WO2024051679A1 - Fak抑制剂及egfr-tki的药物组合及用途 - Google Patents

Fak抑制剂及egfr-tki的药物组合及用途 Download PDF

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WO2024051679A1
WO2024051679A1 PCT/CN2023/116969 CN2023116969W WO2024051679A1 WO 2024051679 A1 WO2024051679 A1 WO 2024051679A1 CN 2023116969 W CN2023116969 W CN 2023116969W WO 2024051679 A1 WO2024051679 A1 WO 2024051679A1
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cancer
inhibitor
pharmaceutically acceptable
growth factor
tyrosine kinase
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French (fr)
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张宝袁
刘学彬
高家明
张平
庞然
王在琪
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应世生物科技(南京)有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/444—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring heteroatom, e.g. amrinone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the invention belongs to the field of medicinal chemistry. Specifically, the present invention relates to focal adhesion kinase (Focal Adhesion Kinase, FAK) inhibitors combined with other drugs to treat tumors.
  • focal adhesion kinase Fecal Adhesion Kinase, FAK
  • ICD Immunogenic cell death
  • ER stress endoplasmic reticulum stress
  • oxidative stress reactive oxygen species stress
  • DAMPs damage-associated molecular patterns
  • High mobility group box 1, HMGB1 High mobility group box 1, HMGB1, etc.
  • This type of DAMPs will be specifically recognized by pattern recognition receptors on Antigen-presenting cells (APCs) in the body, induce the maturation, differentiation and activation of APCs, and gradually present them to immune cells such as effector T cells, thereby It causes immune cells to generate antigen memory, and when tumor cells from the same source are found again, the immune cells will specifically recognize and kill the tumor cells.
  • the new tumor-specific immune response initiated by ICD can increase sensitivity to immune checkpoint inhibitors (Immuno-checkpoint inhibitors, ICI), thereby enhancing the effect of immune checkpoint inhibitors and producing immune memory. Durable antitumor response.
  • FAK also known as protein tyrosine kinase 2 (PTK2), is a non-receptor tyrosine kinase and a key component of the focal adhesion complex. FAK plays an important role in mediating integrin and growth factor signaling to regulate tumor cell invasion, proliferation, and survival.
  • PTK2 protein tyrosine kinase 2
  • EGFR is the abbreviation of Epidermal Growth Factor Receptor.
  • the EGFR gene codes for and makes a receptor protein called epidermal growth factor receptor.
  • the EGFR receptor protein is a transmembrane protein that is divided into three parts: one end of the protein is located outside the cell, one part is located in the cell membrane, and the other end is located within the cell. This allows the EGFR receptor to bind to other proteins outside the cell, called ligands, helping the cell receive the signal and respond to its stimulation.
  • the binding of receptors and ligands is like a key and a lock, so they all have specific binding "partners".
  • EGFR When EGFR binds to a ligand, it attaches to another nearby EGFR receptor and forms a complex (dimer), thereby entering an active state and activating intracellular signaling pathways.
  • EGFR mutations mainly occur in exons 18 to 21, of which deletion mutations in exon 19 and L858R point mutations in exon 21 are the most common mutation types, accounting for 90% of all mutation types.
  • the EGFR receptor protein will be in a continuously activated state, which causes cells to continue to receive proliferation and survival signals, causing excessive cell growth and survival (inability to undergo normal apoptosis), leading to the formation of tumors.
  • Targeted drugs currently on the market for EGFR mutations include: first-generation Icotinib, gefitinib, and erlotinib targeting exon 19 and 21 mutations; second-generation drugs targeting exons 8 and 21.
  • Targeted drugs targeting ALK mutations include: first-generation targeted drugs crizotinib, second-generation targeted drugs ceritinib, alectinib, brigatinib and The third-generation targeted drug lorlatinib, etc.
  • resistance to these targeted drugs mostly appears about 1 year after treatment. Overcoming resistance to targeted drugs, or delaying resistance and improving the possibility of cure are the main goals of drug development.
  • One aspect of the present disclosure provides the use of FAK inhibitors, epidermal growth factor receptor tyrosine kinase inhibitors, and immune checkpoint inhibitors in the preparation of a medicament for treating tumors in a subject.
  • Yet another aspect of the present disclosure provides a pharmaceutical combination product of a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor, and an immune checkpoint inhibitor for use in treating tumors in a subject.
  • Yet another aspect of the present disclosure provides a method of treating tumors, the method comprising providing Such as administering therapeutically effective amounts of FAK inhibitors, epidermal growth factor receptor tyrosine kinase inhibitors and immune checkpoint inhibitors.
  • kit or pharmaceutically acceptable composition which includes: (a) a FAK inhibitor; (b) an epidermal growth factor receptor tyrosine kinase inhibitor; and (c) immune Checkpoint inhibitors.
  • Another aspect of the present disclosure provides the use of a FAK inhibitor and an epidermal growth factor receptor tyrosine kinase inhibitor in the preparation of a medicament for treating tumors, wherein the FAK inhibitor is used to enhance the epidermal growth factor receptor tyrosine kinase inhibitor.
  • Yet another aspect of the present disclosure provides FAK inhibitors for use in the treatment of tumors to enhance immunogenic cell death induced by epidermal growth factor receptor tyrosine kinase inhibitors.
  • Yet another aspect of the present disclosure provides a method of treating tumors, the method comprising administering to a subject in need thereof a therapeutically effective amount of a FAK inhibitor and an epidermal growth factor receptor tyrosine kinase inhibitor, wherein the FAK inhibitor is To enhance immunogenic cell death induced by the epidermal growth factor receptor tyrosine kinase inhibitor.
  • Yet another aspect of the present disclosure provides the use of a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor, and an immune checkpoint inhibitor in the preparation of a medicament for the combined treatment of tumors.
  • Yet another aspect of the present disclosure provides the use of a FAK inhibitor in the preparation of a medicament for the treatment of tumors in combination with an epidermal growth factor receptor tyrosine kinase inhibitor and an immune checkpoint inhibitor.
  • Yet another aspect of the present disclosure provides the use of an epidermal growth factor receptor tyrosine kinase inhibitor in the preparation of a medicament for use in combination with a FAK inhibitor and an immune checkpoint inhibitor to treat tumors.
  • Yet another aspect of the present disclosure provides the use of an immune checkpoint inhibitor in the preparation of a medicament for the treatment of tumors in combination with a FAK inhibitor and an epidermal growth factor receptor tyrosine kinase inhibitor.
  • Yet another aspect of the present disclosure provides the use of a FAK inhibitor in the preparation of a medicament for the treatment of tumors in combination with an epidermal growth factor receptor tyrosine kinase inhibitor and an immune checkpoint inhibitor.
  • Yet another aspect of the present disclosure provides the use of an epidermal growth factor receptor tyrosine kinase inhibitor in the preparation of a medicament for the treatment of tumors in combination with a FAK inhibitor and an immune checkpoint inhibitor.
  • Yet another aspect of the present disclosure provides the use of an immune checkpoint inhibitor in the preparation of a medicament for the treatment of tumors in combination with a FAK inhibitor and an epidermal growth factor receptor tyrosine kinase inhibitor.
  • kits which includes: a FAK inhibitor; and instructions indicating that the FAK inhibitor can be used in combination with an epidermal growth factor receptor tyrosine kinase inhibitor and an immune checkpoint inhibitor. Treat tumors.
  • kits which includes: an epidermal growth factor receptor tyrosine kinase inhibitor; and instructions indicating that the epidermal growth factor receptor tyrosine kinase inhibitor can be used with FAK inhibition. combination of agents and immune checkpoint inhibitors to treat tumors.
  • kits which includes: an immune checkpoint inhibitor; and instructions indicating that the immune checkpoint inhibitor can be used with FAK inhibitors and epidermal growth factor receptor tyrosine kinase inhibition. Combination treatment of tumors.
  • Another aspect of the present disclosure provides a method of treating tumors, comprising administering to a subject in need thereof a therapeutically effective amount of a FAK inhibitor and an epidermal growth factor receptor tyrosine kinase inhibitor.
  • Yet another aspect of the present disclosure provides a pharmaceutical combination product of a FAK inhibitor and an epidermal growth factor receptor tyrosine kinase inhibitor for use in treating tumors in a subject in need thereof.
  • Yet another aspect of the present disclosure provides the use of a FAK inhibitor and an epidermal growth factor receptor tyrosine kinase inhibitor in the preparation of a combination drug for treating tumors.
  • Yet another aspect of the present disclosure provides the use of a FAK inhibitor in the preparation of a medicament for use in combination with an epidermal growth factor receptor tyrosine kinase inhibitor to treat tumors.
  • Yet another aspect of the present disclosure provides the use of an epidermal growth factor receptor tyrosine kinase inhibitor in the preparation of a medicament for use in combination with a FAK inhibitor to treat tumors.
  • Yet another aspect of the present disclosure provides the use of a FAK inhibitor and an epidermal growth factor receptor tyrosine kinase inhibitor in the preparation of a medicament for combined treatment of tumors.
  • Yet another aspect of the present disclosure provides the use of a FAK inhibitor in the preparation of a medicament for the treatment of tumors in combination with an epidermal growth factor receptor tyrosine kinase inhibitor.
  • Yet another aspect of the present disclosure provides the use of an epidermal growth factor receptor tyrosine kinase inhibitor in the preparation of a medicament for the treatment of tumors in combination with a FAK inhibitor.
  • kits which includes: a FAK inhibitor; and instructions indicating that the FAK inhibitor can be used to treat tumors in combination with an epidermal growth factor receptor tyrosine kinase inhibitor.
  • kits which includes: an epidermal growth factor receptor tyrosine kinase inhibitor; and instructions indicating that the epidermal growth factor receptor tyrosine kinase inhibitor can be used with FAK inhibition. Combination treatment of tumors.
  • the FAK inhibitor is IN10018, Defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886 or a pharmaceutically acceptable salt thereof, preferably IN10018, AMP945, Defactinib, or a pharmaceutically acceptable salt thereof.
  • Pharmaceutically acceptable salt enter The first step is preferably IN10018 or a pharmaceutically acceptable salt thereof, especially IN10018 tartrate.
  • the structure of IN10018 is as follows:
  • the Defactinib is also called defatinib, and its CAS number is 1345713-71-4; the CAS number of GSK2256098 is 1224887-10-8.
  • the CAS number of PF-00562271 is 717907-75-0; the CAS number of VS-4718 is 1061353-68-1; the APG-2449 is developed by Ascentage Pharmaceuticals; the CAS number of AMP945 is 1393653- 34-3.
  • the epidermal growth factor receptor tyrosine kinase inhibitor is Gefitinib, Erlotinib, Icotinib, Afatinib, Dacomitinib, Crizotinib, Osimertinib (AZD9291), Almonertinib, Alflutinib (also known as Furmonertinib) , EAI045, JBJ-04-125-02, BLU-945, BLU-701, TQB3804, BBT-176, ES-072, BPI-361175, CH7233163 or their pharmaceutically acceptable salts.
  • the CAS number of Gefitinib is 184475-35-2; the CAS number of Erlotinib is 183321-74-6; the CAS number of Icotinib is 610798-31 -7; the CAS number of Afatinib is 850140-72-6; the CAS number of Crizotinib is 877399-52-5; the CAS number of Osimertinib (AZD9291) is 1421373-65-0; the CAS number of Almonertinib is 1899921-05-1; the CAS number of Alflutinib (also known as Furmonertinib) is 1869057-83-9; EAI045 The CAS number of JBJ-04-125-02 is 2060610-53-7; the CAS number of BLU-945 is 2660250-10-0; BLU-701 is a product of Cable Figure Pharmaceutical Company ( Developed jointly by Blueprint Medicines Corp) and Zai Lab; TQB3804’s CAS number is 2267329
  • the epidermal growth factor receptor tyrosine kinase inhibitor is Osimertinib, Almonertinib, Alflutinib or a pharmaceutically acceptable salt thereof.
  • the epidermal growth factor receptor tyrosine kinase inhibitor is Osimertinib or its drug Scientifically acceptable salt.
  • the epidermal growth factor receptor tyrosine kinase inhibitor is Almonertinib or a pharmaceutically acceptable salt thereof.
  • the epidermal growth factor receptor tyrosine kinase inhibitor is Alflutinib or a pharmaceutically acceptable salt thereof.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody, a PD-1/PD-L1 small molecule inhibitor or a TIGIT inhibitor.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody.
  • the anti-PD-1/PD-L1 antibody is pembrolizumab or tislelizumab. Tislelizumab, Nivolumab, Toripalimab, Atezolizumab, durvalumab, Avelumab, Camrelizumab, Sintilimab, Cemiplimab, envafolimab, BMS-936559, JS003, SHR-1316, GS- 4224, AN-4005 or MX-10181.
  • the immune checkpoint inhibitor is a small molecule inhibitor of PD-1/PD-L1.
  • the small molecule inhibitor of PD-1/PD-L1 is INCB-086550, lazertinib ( Lazertinib), IMMH-010, CA-170, ABSK043 or RRx-001.
  • the immune checkpoint inhibitor is a TIGIT inhibitor.
  • the TIGIT inhibitor is Ociperlimab (Ociperlimab/BGB-A1217), Vibostolimab (Vibostolimab), domvanalimab (AB154 ), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939 , COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof
  • the epidermal growth factor receptor tyrosine kinase inhibitor is Osimertinib or a pharmaceutically acceptable salt thereof
  • the immune checkpoint inhibition The agent is an anti-PD-1/PD-L1 antibody or a PD-1/PD-L1 small molecule inhibitor.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof
  • the epidermal growth factor receptor tyrosine kinase inhibitor is Osimertinib or a pharmaceutically acceptable salt thereof
  • the immune checkpoint inhibition The agent is an anti-PD-1/PD-L1 antibody.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, and the table
  • the epidermal growth factor receptor tyrosine kinase inhibitor is Almonertinib or a pharmaceutically acceptable salt thereof;
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or PD-1/PD-L1 small molecule inhibitor agent.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof
  • the epidermal growth factor receptor tyrosine kinase inhibitor is Almonertinib or a pharmaceutically acceptable salt thereof
  • the immune checkpoint inhibition The agent is an anti-PD-1/PD-L1 antibody.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof
  • the epidermal growth factor receptor tyrosine kinase inhibitor is Alflutinib or a pharmaceutically acceptable salt thereof
  • the immune checkpoint inhibition The agent is an anti-PD-1/PD-L1 antibody or a PD-1/PD-L1 small molecule inhibitor.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof
  • the epidermal growth factor receptor tyrosine kinase inhibitor is Alflutinib or a pharmaceutically acceptable salt thereof
  • the immune checkpoint inhibition The agent is an anti-PD-1/PD-L1 antibody.
  • the FAK inhibitor, the epidermal growth factor receptor tyrosine kinase inhibitor and the immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.
  • the tumor is bladder cancer, breast cancer, cervical cancer, colon cancer (including colorectal cancer), esophageal cancer, esophageal squamous cell carcinoma, head and neck cancer, liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer).
  • melanoma myeloma, rhabdomyosarcoma, inflammatory myofibroblastic tumor, neuroblastoma, pancreatic cancer, prostate cancer, kidney cancer, renal cell carcinoma, sarcoma (including osteosarcoma), skin cancer (including squamous cell carcinoma) (like cell carcinoma), gastric cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal cancer, neuroendocrine cancer, ovarian cancer, salivary gland cancer, spindle cell carcinoma anaplastic large cell lymphoma, anaplastic thyroid carcinoma, non-Hodgkin lymphoma, Hodgkin lymphoma, glioma, or hematological malignancies such as acute myeloid leukemia (AML), acute lymphoblastic leukemia Leukemia (ALL), diffuse large B-cell lympho
  • the tumor is preferably breast cancer, ovarian cancer, colon cancer (including colorectal cancer), lung cancer (including small cell lung cancer and non-small cell lung cancer), melanoma or pancreatic cancer.
  • the tumor is lung cancer (including small cell lung cancer and non-small cell lung cancer) or colon cancer (including colorectal cancer).
  • the tumor is non-small cell lung cancer or colon cancer (including colorectal cancer).
  • Figure 1 shows a white light microscope photo of the cells taken after the lung cancer KPL cells in Example 1 were incubated with drugs for 48 hours.
  • Figure 2a shows the percentage of CRT-positive KPL cells after incubation of lung cancer KPL cells in Example 1 with drugs for 48 hours
  • Figure 2b shows the percentage of Annexin V-positive KPL cells in lung cancer KPL cells in Example 1 after incubation with drugs for 48 hours.
  • Figure 3 shows a white light microscope photo of the cells taken after the lung cancer KPL cells in Example 2 were incubated with drugs for 48 hours.
  • Figure 4a shows the percentage of CRT-positive KPL cells after incubation of lung cancer KPL cells in Example 2 with drugs for 48 hours
  • Figure 4b shows the percentage of Annexin V-positive KPL cells in lung cancer KPL cells in Example 2 after incubation with drugs for 48 hours.
  • Figure 5 shows a white light microscope photo of the cells taken after the lung cancer KPL cells in Example 3 were incubated with drugs for 48 hours.
  • Figure 6a shows the percentage of CRT-positive KPL cells after incubation of lung cancer KPL cells in Example 3 with drugs for 48 hours
  • Figure 6b shows the percentage of Annexin V-positive KPL cells after incubation of lung cancer KPL cells in Example 3 with drugs for 48 hours.
  • Figure 7 shows the early and late apoptosis of cells after FAK silencing and treatment with 0.3nM AZD9291 for 48 hours.
  • Figure 8 shows that combined treatment with 0.3nM AZD9291 for 48 hours after FAK silencing enhanced calreticulin release and exposure.
  • Figure 9 shows the synergistic killing effect of AZD9291 at different doses with 3 ⁇ M IN10018 and 5 ⁇ M IN10018 for 48 hours on non-small cell lung cancer cells HCC827.
  • Figure 10 shows the detection of cell apoptosis after 48 hours of co-administration of 0.3nM AZD9291 and 3 ⁇ M IN10018.
  • Figure 11 shows that the combination of AZD9291 and IN10018 significantly up-regulated the endoplasmic reticulum stress of HCC827.
  • Figure 12 shows that coadministration of AZD9291 with IN10018 enhanced calreticulin release and exposure.
  • Figure 13 shows the changes in tumor volume after administration of different test substances in the colon cancer CT26 cell BALB/c mouse subcutaneous allograft tumor model.
  • Figure 14 shows changes in mouse body weight after administration of different test substances in the colon cancer CT26 cell BALB/c mouse subcutaneous allograft tumor model.
  • Figure 15 shows the changes in tumor volume after administration of different test substances in the colon cancer MC38 cell C57BL/6 mouse subcutaneous allograft tumor model.
  • Figure 16 shows changes in mouse body weight after administration of different test substances in the colon cancer MC38 cell C57BL/6 mouse subcutaneous allograft tumor model.
  • Figure 17a shows the percentage of CRT-positive 4T1 cells after 48 hours of incubation of breast cancer 4T1 cells in Example 8 with drugs
  • Figure 17b shows the percentage of Annexin V-positive 4T1 cells after 48 hours of incubation of breast cancer 4T1 cells in Example 8 with drugs.
  • Figure 18a shows the percentage of CRT-positive 4T1 cells after 48 hours of incubation of breast cancer 4T1 cells in Example 9 with drugs
  • Figure 18b shows the percentage of Annexin V-positive 4T1 cells after 48 hours of incubation of breast cancer 4T1 cells in Example 9 with drugs.
  • Figure 19a shows the percentage of CRT-positive 4T1 cells after 48 hours of incubation of breast cancer 4T1 cells in Example 10 with drugs
  • Figure 19b shows the percentage of Annexin V-positive 4T1 cells after 48 hours of incubation of breast cancer 4T1 cells in Example 10 with drugs.
  • FAK inhibitor refers to a potent inhibitor of FAK, which may be suitable for mammals, especially humans.
  • the FAK inhibitor is IN10018, Defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886 or a pharmaceutically acceptable salt thereof, and the structure of IN10018 is as follows:
  • the Defactinib is also called defatinib, and its CAS number is 1345713-71-4; the CAS number of GSK2256098 is 1224887-10-8.
  • the CAS number of PF-00562271 is 717907-75-0; the CAS number of VS-4718 is 1061353-68-1; the APG-2449 is developed by Ascentage Pharmaceuticals; the CAS number of AMP945 is 1393653- 34-3.
  • the FAK inhibitor is preferably IN10018, AMP945, Defactinib or a pharmaceutically acceptable salt thereof. In some preferred embodiments, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, Especially IN10018 tartrate.
  • epidermal growth factor receptor tyrosine kinase inhibitor EGFR-TKI
  • epidermal growth factor receptor tyrosine kinase inhibitors include, but are not limited to, gefitinib, erlotinib Erlotinib, Icotinib, Afatinib, Dacomitinib, Crizotinib, Osimertinib (AZD9291), Ametinib (Almonertinib), Alflutinib (also known as Furmonertinib), EAI045, JBJ-04-125-02, BLU-945, BLU-701, TQB3804, BBT-176, ES-072, BPI-361175, CH7233163 or their pharmaceutically acceptable salts.
  • the CAS number of Gefitinib is 184475-35-2; the CAS number of Erlotinib is 183321-74-6; the CAS number of Icotinib is 610798-31 -7; the CAS number of Afatinib is 850140-72-6; the CAS number of Dacomitinib is 1110813-31-4; the CAS number of Crizotinib is 877399- 52-5; the CAS number of Osimertinib (AZD9291) is 1421373-65-0; the CAS number of Almonertinib (Almonertinib) is 1899921-05-1; Alflutinib (Alflutinib, also known as Fumei) The CAS number of Furmonertinib is 1869057-83-9; the CAS number of EAI045 is 1942114-09-1; the CAS number of JBJ-04-125-02 is 2060610-53-7; the CAS number of BLU-945
  • immune checkpoint inhibitor refers to drugs that can improve the activity of the immune system by regulating immune checkpoint pathways (such as PD-1, TIGIT, CTLA-4, LAG-3, TIM-3, etc.).
  • the immune checkpoint inhibitor is an antagonist of the PD-1/PD-L1 (programmed cell death protein 1) pathway (also known as a "PD-1 inhibitor”) or a TIGIT inhibitor.
  • PD-1 inhibitors are also referred to as PD-1/PD-L1 inhibitors in this disclosure.
  • the PD-1/PD-L1 inhibitor is a PD-1/PD-L1 antibody, including but not limited to pembrolizumab (Keytruda) , Tislelizumab (Tislelizumab/Baizian), Nivolumab (Nivolumab), Toripalimab (Toripalimab/Tuoyi), Duvalumab (Infinifer/Duvalumab) /durvalumab), avelumab (Avelumab/Bavencio), atezolizumab (MPDL3280A/Atezolizumab/Tecentriq), BMS-936559 (fully human anti-PD-L1 IgG4 monoclonal antibody), GS-4224, AN-4005 or MX-10181.
  • pembrolizumab Keytruda
  • Tislelizumab Tislelizumab/Baizian
  • Nivolumab Nivolumab
  • the PD-1 inhibitor is toripalimab. In some embodiments, PD-1 inhibitors are used to treat Treatment object. In some embodiments, PD-1 is human PD-1.
  • PD-1/PD-L1 inhibitors also include PD-1/PD-L1 small molecule inhibitors, such as INCB-086550, Lazertinib, IMMH-010, CA-170, ABSK043 or RRx-001.
  • TIGIT also known as WUCAM, Vstm3, VSIG9 is a receptor of the Ig superfamily and a new immune checkpoint after PD-1/PD-L1.
  • the immune checkpoint inhibitor is a TIGIT inhibitor, including but not limited to Ociperlimab (Ociperlimab/BGB-A1217), Vibostolimab (Vibostolimab) ), domvanalimab(AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M- 6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, HB0036 or IBI-321.
  • TIGIT inhibitors are used to treat human subjects. To avoid ambiguity, antibodies in this article include double antibodies.
  • pharmaceutical combination or “pharmaceutical combination product” may refer to a fixed combination in the form of a dosage unit (for example, all active pharmaceutical ingredients are present in one dosage form) or a kit of products for combined administration. Refers to the combination of a drug and instructions indicating that the drug can be used in combination with one or more other drugs.
  • combination therapy or “combination drug” refers to the combination of one drug with one or more other drugs to treat a disease, including the combination of one drug with another one or more drugs. This also includes the combination of a drug and instructions indicating that the drug can be used in combination with one or more other drugs.
  • “Simultaneous or sequential administration” in this application refers to two or more drugs administered simultaneously or at a certain time interval within one administration cycle (for example, within 4 weeks, within 3 weeks, within 2 weeks, within 1 week, or within 24 hours). Administered sequentially, the methods of drug administration (such as oral, intravenous, intramuscular or subcutaneous administration, etc.) may be the same or different, and the administration frequency/period of two or more drugs may be the same or different. When the treatment methods, products or uses of the present disclosure involve two drugs, the two drugs can be administered separately at the same time or at certain intervals.
  • the three drugs can be administered at the same time point, or two drugs can be administered at one time point and one drug can be administered at another time point, or all Each of the three drugs was administered at different time points.
  • the PD-1/PD-L1 inhibitor is administered intravenously (eg, as an intravenous infusion) or subcutaneously, or orally.
  • the PD-1/PD-L1 inhibitor is administered as an intravenous infusion.
  • the TIGIT inhibitor is administered intravenously (e.g., as an intravenous infusion) or subcutaneously Administer or take by mouth.
  • the TIGIT inhibitor is administered as an intravenous infusion.
  • immune checkpoint inhibitors to treat cancer depends on the presence of tumor antigen-specific T cells within tumor tissue. This requires that tumor tissues express antigens that distinguish themselves from their non-transformed counterparts, for example, through novel protein products called neoantigens. Tumor neoantigen burden is closely related to immunogenicity and sensitivity (e.g., to checkpoint inhibitor therapies), meaning that less immunogenic tumors should be largely resistant to these agents. Therapies designed to release tumor antigens that can be taken up by APCs, such as those that induce immunogenic cell death (ICD), may promote effective anti-tumor immunity, especially when further combined with checkpoint inhibitors.
  • ICD immunogenic cell death
  • treatment refers to the administration of one or more pharmaceutical substances to a subject suffering from a disease or having symptoms of said disease in order to cure, alleviate, alleviate, alter, treat, ameliorate, ameliorate or affect said disease.
  • Disease or symptoms of said disease In some embodiments, the disease is tumor or cancer.
  • tumor refers to abnormal lesions formed by the body's abnormal growth caused by various tumorigenic factors. Cells in local tissues lose normal control of their growth at the genetic level, resulting in abnormal proliferation of their clonal types.
  • the tumors include, but are not limited to: bladder cancer, breast cancer, cervical cancer, colon cancer (including colorectal cancer), esophageal cancer, esophageal squamous cell carcinoma, head and neck cancer, liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer).
  • small cell lung cancer small cell lung cancer
  • melanoma myeloma, rhabdomyosarcoma, inflammatory myofibroblastic tumor, neuroblastoma, pancreatic cancer, prostate cancer, kidney cancer, renal cell carcinoma, sarcoma (including osteosarcoma), skin cancer (including Squamous cell carcinoma), gastric cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal cancer, neuroendocrine cancer, ovarian cancer, salivary gland cancer, spindle cell carcinoma Metastases, anaplastic large cell lymphoma, anaplastic thyroid carcinoma, non-Hodgkin lymphoma, Hodgkin lymphoma, glioma, and hematological malignancies such as acute myeloid leukemia (AML), acute lymphoblastic leukemia Leukemia (ALL), diffuse large B
  • the term "subject” or “subject” refers to mammals and non-mammals.
  • Mammal means any member of the class Mammalia, including but not limited to: humans; non-human primates, such as Chimpanzees and other ape and monkey species; farm animals, such as cattle, horses, sheep, goats, and pigs; domestic animals, such as rabbits, dogs, and cats; laboratory animals, including rodents, such as rats, mice, and guinea pigs ;etc. Examples of non-mammals include, but are not limited to, birds.
  • the term "subject” does not identify a specific age or gender. In some embodiments, the subject is a human.
  • pharmaceutically acceptable means nontoxic, biologically tolerable, and suitable for administration to a subject.
  • pharmaceutically acceptable salts refers to nontoxic, biologically tolerable acid addition salts suitable for administration to a subject, including, but not limited to, acid addition salts with inorganic acids. , such as hydrochlorides, hydrobromides, carbonates, bicarbonates, phosphates, sulfates, sulfites, nitrates, etc.; and acid addition salts formed with organic acids, such as formates, acetates, etc.
  • Acid malate, maleate, fumarate, tartrate, succinate, citrate, lactate, methanesulfonate, p-toluenesulfonate, 2-hydroxyethanesulfonate , benzoate, salicylate, stearate and salts formed with alkanedicarboxylic acids of the formula HOOC-(CH 2 ) n -COOH (where n is 0-4), etc.
  • pharmaceutically acceptable acid addition salts can be obtained by dissolving the free base in a suitable solvent and treating the solution with an acid according to conventional procedures for preparing acid addition salts from basic compounds.
  • suitable solvent for preparing acid addition salts from basic compounds.
  • One skilled in the art will be able to identify, without undue experimentation, various synthetic methods that can be used to prepare nontoxic pharmaceutically acceptable acid addition salts.
  • composition means that it must be chemically and/or toxicologically compatible with the other ingredients including the formulation, and/or with the subject to be treated therewith.
  • therapeutically effective amount refers to an amount generally sufficient to produce a beneficial therapeutic effect in a subject. Conventional influencing factors (e.g., mode of administration, pharmacokinetics of the compound, severity and course of disease, subject's medical history, subject's health condition, subject degree of response to drugs, etc.) to determine the therapeutically effective dose of the present invention.
  • inhibitor refers to a decrease in the baseline activity of a biological activity or process.
  • kit used in this article refers to a box used to contain chemical reagents for detecting chemical components, drug residues, virus types, etc.
  • the kit of the present invention may include (i) one, two or three of FAK inhibitors, epidermal growth factor receptor tyrosine kinase inhibitors and immune checkpoint inhibitors; and (ii) instructions, The instructions state that FAK inhibitors, epidermal growth factor receptor tyrosine kinase inhibitors, and immune checkpoint inhibitors can be used to treat tumors in a subject.
  • the kit includes (i) a FAK inhibitor; and (ii) instructions for use of a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor, and an immune checkpoint inhibitor to treat tumors in subjects.
  • the kit includes (i) an epidermal growth factor receptor tyrosine kinase inhibitor; and (ii) instructions indicating that the FAK inhibitor, epidermal growth factor receptor tyrosine kinase, can be used inhibitors and immune checkpoint inhibitors to treat tumors in subjects.
  • the kit includes (i) an immune checkpoint inhibitor; and (ii) instructions for using a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor, and an immune checkpoint inhibitor. Inhibitors are used to treat tumors in subjects.
  • the kit includes (i) a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor, and an immune checkpoint inhibitor; and (ii) instructions indicating that the FAK inhibitor can be used , epidermal growth factor receptor tyrosine kinase inhibitors, and immune checkpoint inhibitors to treat tumors in subjects.
  • the kit includes (i) a FAK inhibitor; and (ii) instructions indicating that the FAK inhibitor and the epidermal growth factor receptor tyrosine kinase inhibitor can be used to treat tumors in a subject .
  • the kit includes (i) an epidermal growth factor receptor tyrosine kinase inhibitor; and (ii) instructions for using the epidermal growth factor receptor tyrosine kinase inhibitor and FAK The inhibitor treats tumors in the subject.
  • kits may include a first container including at least one dose of a drug including a FAK inhibitor, a second container including at least one dose of epidermal growth factor receptor, a third container and a package insert. a tyrosine kinase inhibitor, the third container includes at least one dose of an immune checkpoint inhibitor drug, and the package insert includes instructions for using the drug to treat the subject's tumor.
  • the first container, the second container, and the third container may contain the same or different shapes (eg, vials, syringes, and bottles) and/or materials (eg, plastic or glass). Kits may also include other materials that may aid in administering the medication, such as diluents, filters, IV bags and tubing, needles, and syringes.
  • FAK inhibitor secretoryl growth factor receptor tyrosine kinase inhibitor
  • immune checkpoint inhibitor an effective amount also depends on the degree, severity and type of cell proliferation. A technician will be able to determine the appropriate dosage based on these and other factors.
  • FAK inhibitors can be administered in a suitable manner, such as oral, intravenous, intramuscular, or subcutaneous administration.
  • the drug may be administered orally with a pharmaceutically acceptable carrier such as an inert diluent or an absorbable edible carrier. They can be enclosed in hard- or soft-shell gelatin capsules, compressed into tablets, or can be mixed directly with the patient's food.
  • a pharmaceutically acceptable carrier such as an inert diluent or an absorbable edible carrier. They can be enclosed in hard- or soft-shell gelatin capsules, compressed into tablets, or can be mixed directly with the patient's food.
  • the drug may be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, lozenges, capsules, elixirs, suspensions, syrups, or wafers. Tablets, lozenges, pills, capsules, etc.
  • a binder such as tragacanth, acacia, corn starch or gelatin
  • an excipient such as dicalcium phosphate
  • a disintegrant such as corn starch, potato starch , alginic acid, etc.
  • lubricants such as magnesium stearate
  • sweeteners such as sucrose, fructose, lactose or aspartame; or flavoring agents.
  • solutions of the drug may be prepared in water, optionally mixed with a nontoxic surfactant.
  • Exemplary pharmaceutical dosage forms for injection or infusion include sterile aqueous solutions, dispersions, or sterile powders containing the active ingredient suitable for the extemporaneous preparation of sterile injection or infusion solutions or dispersions. Regardless, the final dosage form should be sterile, fluid, and stable under the conditions of manufacture and storage.
  • Sterile injectable solutions can be prepared by incorporating the required amount of the drug in an appropriate solvent with various other ingredients required from above, followed by filtered sterilization.
  • the preferred methods of preparation may be vacuum drying and freeze-drying techniques, which produce a powder of the active ingredient plus any other desired ingredients present after previous sterile filtration.
  • the amount of FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor, or immune checkpoint inhibitor required for treatment may vary not only with the specific agent selected, but also with the route of administration, disease being treated varies with the nature of the disease as well as the age and condition of the patient and can ultimately be determined at the discretion of the attending physician or clinician. In general, however, dosages may range from about 0.1 to about 50 mg/kg of body weight per day.
  • the FAK inhibitors are administered at doses ranging from 5 mg/day to 300 mg/day in adults.
  • IN10018, or a pharmaceutically acceptable salt thereof is administered at a dose of 5 mg/day to 100 mg/day in adults.
  • IN10018, or a pharmaceutically acceptable salt thereof is administered in adults. In adults doses of 25 mg/day to 100 mg/day are administered as free base.
  • the epidermal growth factor receptor tyrosine kinase inhibitor is administered in a dosage range of 2-250 mg per day in adults.
  • Osimertinib or a pharmaceutically acceptable salt thereof is administered in adults at a dose of 2-250 mg, for example, 80 mg per day, based on Osimertinib; Almonertinib or a pharmaceutically acceptable salt thereof is administered in adults.
  • a dose of 2-250 mg, such as 110 mg, is administered per day, the dose being calculated as Almonertinib; Alflutinib or a pharmaceutically acceptable salt thereof to form
  • a dose of 2-250 mg, for example 80 mg, of alflutinib per day is administered.
  • the immune checkpoint inhibitor is administered at a dose of 2-10 mg/kg or 50-1200 mg in adults every 2 to 3 weeks. In a specific embodiment, the immune checkpoint inhibitor is administered at a dose of 3-10 mg/kg or 100-1200 mg in adults, once every 2 weeks to 3 weeks.
  • the present disclosure also discloses the following:
  • FAK inhibitors said epidermal growth factor receptor tyrosine kinase inhibitors and immune checkpoint inhibitors, methods for treating tumors in a subject.
  • the epidermal growth factor receptor tyrosine kinase inhibitor is Gemphil Gefitinib, Erlotinib, Icotinib, Afatinib, Dacomitinib,
  • the FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 1-3, wherein the epidermal growth factor receptor tyrosine kinase inhibitor It is Osimertinib, Almonertinib, Alflutinib or a pharmaceutically acceptable salt thereof.
  • the immune checkpoint inhibitor is anti-PD-1/ PD-L1 antibody
  • the anti-PD-1/PD-L1 antibody is pembrolizumab, tislelizumab, nivolumab, terep Toripalimab, Atezolizumab, durvalumab, Avelumab, Camrelizumab, Sintilimab ), cemiplimab, envolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.
  • the immune checkpoint inhibitor is a PD-1/PD-L1 small molecule inhibitor
  • the PD- 1/PD-L1 small molecule inhibitors are INCB-086550, Lazertinib, IMMH-010, CA-170, ABSK043 or RRx-001.
  • the FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 1-8, wherein the immune checkpoint inhibitor is a TIGIT inhibitor, further , the TIGIT inhibitors are Ociperlimab (Ociperlimab/BGB-A1217), Vibostolimab (Vibostolimab), domvanalimab (AB154), Tiragolumab (Tiragolumab), Belrestotug, Attilizine Monoclonal antibody (Etigilimab), ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.
  • TIGIT inhibitors are
  • the FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 1-4, wherein the FAK inhibitor is IN10018 or pharmaceutically acceptable thereof salt, the epidermal growth factor receptor tyrosine kinase inhibitor is Osimertinib or a pharmaceutically acceptable salt thereof; the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or PD-1/PD -L1 small molecule inhibitors, especially anti-PD-1/PD-L1 antibodies.
  • the FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 1-4, wherein the FAK inhibitor is IN10018 or pharmaceutically acceptable thereof salt, the epidermal growth factor receptor tyrosine kinase inhibitor is Almonertinib or a pharmaceutically acceptable salt thereof; the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or PD-1/PD -L1 small molecule inhibitors, especially anti-PD-1/PD-L1 antibodies.
  • the FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 1-4, wherein the FAK inhibitor is IN10018 or pharmaceutically acceptable thereof salt, the epidermal growth factor receptor tyrosine kinase inhibitor is Alflutinib or a pharmaceutically acceptable salt thereof; the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or PD-1/PD -L1 small molecule inhibitors, especially anti-PD-1/PD-L1 antibodies.
  • the tumor is bladder cancer, breast cancer, or cervical cancer.
  • colon cancer including colorectal cancer
  • esophageal cancer including esophageal squamous cell carcinoma, head and neck cancer
  • liver cancer including lung cancer (including small cell lung cancer and non-small cell lung cancer), melanoma, myeloma, rhabdomyosarcoma, inflammatory myofibroblast Cytoma, neuroblastoma, pancreatic cancer, prostate cancer, kidney cancer, renal cell carcinoma, sarcoma (including osteosarcoma), skin cancer (including squamous cell carcinoma), gastric cancer, testicular cancer, thyroid cancer, uterine cancer, mesenchymal cancer Dermatoma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal carcinoma
  • kits or pharmaceutically acceptable composition comprising:
  • kits or composition of embodiment 18, wherein the FAK inhibitor is IN10018, Defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886 or pharmaceutically acceptable ones thereof
  • the salt is preferably IN10018, AMP945, Defactinib, or a pharmaceutically acceptable salt thereof, and further preferably IN10018 or a pharmaceutically acceptable salt thereof, especially IN10018 tartrate.
  • the structure of IN10018 is as follows:
  • kits or composition of embodiments 18-19, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is Gefitinib, Erlotinib, Ek Icotinib, Afatinib, Dacomitinib, Crizotinib, Osimertinib (AZD9291), Almonertinib, Aflotinib Alflutinib (also known as Furmonertinib), EAI045, JBJ-04-125-02, BLU-945, BLU-701, TQB3804, BBT-176, ES-072, BPI-361175, CH7233163 or other Pharmaceutically acceptable salt.
  • the epidermal growth factor receptor tyrosine kinase inhibitor is Gefitinib, Erlotinib, Ek Icotinib, Afatinib, Dacomitinib, Crizotinib, Osimertinib (AZD9291), Almonertinib,
  • kits or composition of any one of embodiments 18-20, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is Osimertinib, Almonertinib, Alflutinib or a pharmaceutically acceptable salt thereof.
  • the long factor receptor tyrosine kinase inhibitor is Osimertinib or a pharmaceutically acceptable salt thereof.
  • composition of any one of embodiments 18-21, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is Alflutinib or a pharmaceutically acceptable salt thereof.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody
  • the anti-PD-1/PD -L1 antibodies are pembrolizumab, Tislelizumab, Nivolumab, Toripalimab, Atezolizumab , durvalumab, Avelumab, Camrelizumab, Sintilimab, Cemiplimab, Envolimab Anti-(envafolimab), BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.
  • TIGIT inhibitor is Ociperlimab /BGB-A1217), Vibostolimab, domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS
  • kits or composition of embodiment 33, wherein the tumor is lung cancer (including small cell lung cancer and non-small cell lung cancer) or colon cancer (including colorectal cancer), especially non-small cell lung cancer or colon cancer (including colorectal cancer).
  • a method of treating a tumor in a subject comprising administering to the subject a therapeutically effective amount of a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor, and an immune checkpoint inhibitor.
  • epidermal growth factor receptor tyrosine kinase inhibitor is Osimertinib, Almonertinib, Alflutinib or a pharmaceutically acceptable salt thereof.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody, a PD-1/PD-L1 small molecule inhibitor, or a TIGIT inhibitor.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody
  • the anti-PD-1/PD-L1 antibody is Pembrolizumab, Tislelizumab, Nivolumab, Toripalimab, Atezolizumab, Duvalumab Anti-(durvalumab), Avelumab, Camrelizumab, Sintilimab, Cemiplimab, envolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.
  • the immune checkpoint inhibitor is a PD-1/PD-L1 small molecule inhibitor
  • the PD- 1/PD-L1 small molecule inhibitors are INCB-086550, Lazertinib, IMMH-010, CA-170, ABSK043 or RRx-001.
  • the immune checkpoint inhibitor is a TIGIT inhibitor
  • the TIGIT inhibitor is Ociperlimab/BGB-A1217 ), Vibostolimab, domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX- 301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof
  • the epidermal growth factor receptor tyrosine kinase inhibitor is Osimertinib or a pharmaceutically acceptable salt thereof.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or a PD-1/PD-L1 small molecule inhibitor, especially an anti-PD-1/PD-L1 antibody.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof
  • the epidermal growth factor receptor tyrosine kinase inhibitor is Almonertinib or a pharmaceutically acceptable salt thereof.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or a PD-1/PD-L1 small molecule inhibitor, especially an anti-PD-1/PD-L1 antibody.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof
  • the epidermal growth factor receptor tyrosine kinase inhibitor is Alflutinib or a pharmaceutically acceptable salt thereof.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or a PD-1/PD-L1 small molecule inhibitor, especially an anti-PD-1/PD-L1 antibody.
  • the tumor is bladder cancer, breast cancer, cervical cancer, colon cancer (including colorectal cancer), esophageal cancer, esophageal squamous cell carcinoma, head and neck cancer Cancer, liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer), melanoma, myeloma, transverse Stroyosarcoma, inflammatory myofibroblastic tumor, neuroblastoma, pancreatic cancer, prostate cancer, kidney cancer, renal cell carcinoma, sarcoma (including osteosarcoma), skin cancer (including squamous cell carcinoma), gastric cancer, testicular cancer , Thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal cancer, neuroendocrine cancer, ovarian cancer, salivary gland cancer, metastases caused by spindle cell carcinoma, an
  • tumor is lung cancer (including small cell lung cancer and non-small cell lung cancer) or colon cancer (including colorectal cancer), especially non-small cell lung cancer or Colon cancer (including colorectal cancer).
  • lung cancer including small cell lung cancer and non-small cell lung cancer
  • colon cancer including colorectal cancer
  • non-small cell lung cancer or Colon cancer including colorectal cancer
  • FAK inhibitors for use in methods of treating tumors by increasing immunogenic cell death in a subject.
  • the epidermal growth factor receptor tyrosine kinase inhibitor is Gemfil Gefitinib, Erlotinib, Icotinib, Afat
  • the FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 52-58, wherein the immune checkpoint inhibitor is anti-PD-1/ PD-L1 antibodies, PD-1/PD-L1 small molecule inhibitors or TIGIT inhibitors.
  • the FAK inhibitor is anti-PD-1/ PD-L1 antibody
  • the anti-PD-1/PD-L1 antibody is pembrolizumab, tislelizumab, nivolumab, terep Toripalimab, Atezolizumab, durvalumab, Avelumab, Camrelizumab, Sintilimab ), cemiplimab, envolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.
  • the immune checkpoint inhibitor is a PD-1/PD-L1 small molecule inhibitor
  • the PD- 1/PD-L1 small molecule inhibitors are INCB-086550, Lazertinib, IMMH-010, CA-170, ABSK043 or RRx-001.
  • the TIGIT inhibitors are Ociperlimab (Ociperlimab/BGB-A1217), Vibostolimab (Vibostolimab), domvanalimab (AB154), Tiragolumab (Tiragolumab), Belrestotug , Etigilimab, ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.
  • the FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 52-55, wherein the FAK inhibitor is IN10018 or pharmaceutically acceptable thereof salt, the epidermal growth factor receptor tyrosine kinase inhibitor is Osimertinib or a pharmaceutically acceptable salt thereof; the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or PD-1/PD -L1 small molecule inhibitors, especially anti-PD-1/PD-L1 antibodies.
  • the FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 52-55, wherein the FAK inhibitor is IN10018 or pharmaceutically acceptable thereof salt, the epidermal growth factor receptor tyrosine kinase inhibitor is Almonertinib or a pharmaceutically acceptable salt thereof; the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or PD-1/PD -L1 small molecule inhibitors, especially anti-PD-1/PD-L1 antibodies.
  • the FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 52-55, wherein the FAK inhibitor is IN10018 or pharmaceutically acceptable thereof salt, the epidermal growth factor receptor tyrosine kinase inhibitor is Alflutinib or a pharmaceutically acceptable salt thereof; the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or PD-1/PD -L1 small molecule inhibitors, especially anti-PD-1/PD-L1 antibodies.
  • the tumor is bladder cancer, breast cancer, cervical cancer, Colon cancer (including colorectal cancer), esophageal cancer, esophageal squamous cell carcinoma, head and neck cancer, liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer), melanoma, myeloma, rhabdomyosarcoma, inflammatory myofibroblast tumor, neuroblastoma, pancreatic cancer, prostate cancer, kidney cancer, renal cell carcinoma, sarcoma (including osteosarcoma), skin cancer (including squamous cell carcinoma), gastric cancer, testicular cancer, thyroid cancer, Uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal cancer, neuroendocrine
  • the tumor is lung cancer (including small cell lung cancer and non-small cell lung cancer).
  • cell lung cancer or colon cancer (including colorectal cancer), especially non-small cell lung cancer or colon cancer (including colorectal cancer).
  • a method of treating tumors by increasing immunogenic cell death in a subject comprises administering to said subject a therapeutically effective amount of a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor, and Immune checkpoint inhibitors.
  • the FAK inhibitor is IN10018, Defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886 or a pharmaceutically acceptable salt thereof, preferably IN10018, AMP945, Defactinib, or a pharmaceutically acceptable salt thereof, and further preferably IN10018 or a pharmaceutically acceptable salt thereof, especially IN10018 tartrate.
  • the structure of IN10018 is as follows:
  • epidermal growth factor receptor tyrosine kinase inhibitor is Gefitinib, Erlotinib, Icotinib ), Afatinib, Dacomitinib, Crizotinib, Osimertinib (AZD9291), Almonertinib, Alflutinib, Also known as Furmonertinib), EAI045, JBJ-04-125-02, BLU-945, BLU-701, TQB3804, BBT-176, ES-072, BPI-361175, CH7233163 or other pharmaceutically acceptable of salt.
  • epidermal growth factor receptor tyrosine kinase inhibitor is Osimertinib, Almonertinib, Alflutinib, or a pharmaceutically acceptable salt thereof.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody, a PD-1/PD-L1 small molecule inhibitor, or a TIGIT inhibitor .
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody
  • the anti-PD-1/PD-L1 antibody is Pembrolizumab, Tislelizumab, Nivolumab, Toripalimab, Atezolizumab, Duvalumab Anti-durvalumab, Avelumab, Camrelizumab, Sintilimab, Cemiplimab, envafolimab , BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.
  • the immune checkpoint inhibitor is a PD-1/PD-L1 small molecule inhibitor
  • the PD- 1/PD-L1 small molecule inhibitors are INCB-086550, Lazertinib, IMMH-010, CA-170, ABSK043 or RRx-001.
  • the immune checkpoint inhibitor is a TIGIT inhibitor
  • the TIGIT inhibitor is Ociperlimab/BGB-A1217 ), Vibostolimab, domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX- 301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, and the epidermal growth factor receptor tyrosine kinase inhibitor It is Osimertinib or a pharmaceutically acceptable salt thereof;
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or a PD-1/PD-L1 small molecule inhibitor, especially an anti-PD-1/PD- L1 antibodies.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof
  • the epidermal growth factor receptor tyrosine kinase inhibitor is Almonertinib or a pharmaceutically acceptable salt thereof.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or a PD-1/PD-L1 small molecule inhibitor, especially an anti-PD-1/PD-L1 antibody.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof
  • the epidermal growth factor receptor tyrosine kinase inhibitor is Alflutinib or a pharmaceutically acceptable salt thereof.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or a PD-1/PD-L1 small molecule inhibitor, especially an anti-PD-1/PD-L1 antibody.
  • the tumor is bladder cancer, breast cancer, cervical cancer, colon cancer (including colorectal cancer), esophageal cancer, esophageal squamous cell carcinoma, head and neck cancer Cancer, liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer), melanoma, myeloma, rhabdomyosarcoma, inflammatory myofibroblastic tumor, neuroblastoma, pancreatic cancer, prostate cancer, kidney cancer, renal cell carcinoma , Sarcoma (including osteosarcoma), skin cancer (including squamous cell carcinoma), gastric cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal cancer, neuroendocrine cancer, Ovarian cancer, salivary gland cancer, metastases from spindle cell carcinoma, ana
  • AML Acute myeloid leukemia
  • ALL acute lymphoblastic leukemia
  • DLBCL diffuse large B-cell lymphoma
  • FL follicular lymphoma
  • CLL chronic lymphocytic leukemia
  • CML chronic myelogenous leukemia
  • the tumor is breast cancer, ovarian cancer, colon cancer (including colorectal cancer), lung cancer (including small cell lung cancer and non-small cell lung cancer), melanoma and pancreatic cancer.
  • the tumor is lung cancer (including small cell lung cancer and non-small cell lung cancer) or colon cancer (including colorectal cancer), especially non-small cell lung cancer or colon cancer (including colorectal cancer).
  • FAK inhibitor in the preparation of a medicament for treating tumors in a subject, wherein the FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor and an immune checkpoint inhibitor are combined
  • the agent is administered to the subject.
  • an immune checkpoint inhibitor in the preparation of a medicament for treating tumors in a subject, wherein a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor, and the immune checkpoint inhibitor are administered to the Described object.
  • FAK inhibitors Use of FAK inhibitors, epidermal growth factor receptor tyrosine kinase inhibitors and immune checkpoint inhibitors in the preparation of combination drugs for treating tumors.
  • FAK inhibitor in the preparation of a drug for the treatment of tumors in combination with an epidermal growth factor receptor tyrosine kinase inhibitor and an immune checkpoint inhibitor.
  • FAK inhibitors Use of FAK inhibitors, epidermal growth factor receptor tyrosine kinase inhibitors and immune checkpoint inhibitors in the preparation of drugs for combined treatment of tumors.
  • FAK inhibitor in the preparation of a medicament for the treatment of tumors in combination with an epidermal growth factor receptor tyrosine kinase inhibitor and an immune checkpoint inhibitor.
  • the salt is preferably IN10018, AMP945, Defactinib, or a pharmaceutically acceptable salt thereof, and further preferably IN10018 or a pharmaceutically acceptable salt thereof, especially IN10018 tartrate.
  • the structure of IN10018 is as follows:
  • epidermal growth factor receptor tyrosine kinase inhibitor is Gefitinib, Erlotinib, Icoti Icotinib, Afatinib, Dacomitinib, Crizotinib, Osimertinib (AZD9291), Almonertinib, Aflotinib (Alflutinib, also known as Furmonertinib), EAI045, JBJ-04-125-02, BLU-945, BLU-701, TQB3804, BBT-176, ES-072, BPI-361175, CH7233163 or its pharmaceutical with an acceptable salt.
  • the epidermal growth factor receptor tyrosine kinase inhibitor is Gefitinib, Erlotinib, Icoti Icotinib, Afatinib, Dacomitinib, Crizotinib, Osimertinib (AZD9291), Almonertinib, Aflotinib (
  • epidermal growth factor receptor tyrosine kinase inhibitor is Osimertinib, Almonertinib, Alflutinib or a pharmaceutically acceptable salt thereof.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody
  • the anti-PD-1/PD-L1 antibody is Pembrolizumab, Tislelizumab, Nivolumab, Toripalimab, Atezolizumab, Duvalumab Anti-durvalumab, Avelumab, Camrelizumab, Sintilimab, Cemiplimab, envafolimab , BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.
  • the immune checkpoint inhibitor is a small molecule inhibitor of PD-1/PD-L1.
  • the small molecule inhibitor of PD-1/PD-L1 is INCB-086550, Lazertinib, IMMH-010, CA-170, ABSK043 or RRx-001.
  • the immune checkpoint inhibitor is a TIGIT inhibitor
  • the TIGIT inhibitor is Ociperlimab/BGB-A1217 ), Vibostolimab, domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX- 301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or a PD-1/PD-L1 small molecule inhibitor, especially an anti-PD-1/PD-L1 antibody.
  • Pharmaceutically acceptable salt; the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or a PD-1/PD-L1 small molecule inhibitor, especially an anti-PD-1/PD-L1 antibody.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or a PD-1/PD-L1 small molecule inhibitor, especially an anti-PD-1/PD-L1 antibody.
  • the tumor is bladder cancer, breast cancer, cervical cancer, colon cancer (including colorectal cancer), esophageal cancer, esophageal squamous cell carcinoma, head and neck cancer Cancer, liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer), melanoma, myeloma, rhabdomyosarcoma, inflammatory myofibroblastic tumor, neuroblastoma, pancreatic cancer, prostate cancer, kidney cancer, renal cell carcinoma , Sarcoma (including osteosarcoma), skin cancer (including squamous cell carcinoma), gastric cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal cancer, Neuroendocrine cancer, ovarian cancer, salivary gland cancer, metastases from spindle cell carcinoma, ana
  • any one of embodiments 86-111, wherein the tumor is lung cancer (including small cell lung cancer and non-small cell lung cancer) or colon cancer (including colorectal cancer), especially non-small cell lung cancer or colon cancer (including colorectal cancer).
  • lung cancer including small cell lung cancer and non-small cell lung cancer
  • colon cancer including colorectal cancer
  • non-small cell lung cancer or colon cancer including colorectal cancer
  • Example 1 Study of IN10018 and AZD9291 in lung cancer KPL cells
  • KPL cells (Institute of Cell Science, Chinese Academy of Sciences) were cultured with RPMI 1640 (Shanghai Yuanpei, classification number: L210KJ, batch number: F210916) + 10% FBS (Gibco, classification number: 10099-141c, batch number: 2158737cp), and passaged twice.
  • four groups were set up. The first group was the negative control group and culture medium was added. The second group was IN10018 at a concentration of 10 ⁇ M. The third group was AZD9291 at a concentration of 10 ⁇ M. The fourth group was a combination of IN10018 and AZD9291.
  • the use groups are IN10018 (10 ⁇ M) and AZD9291 (10 ⁇ M) respectively. Mix the drugs and incubate in a 5% CO2 incubator at 37°C for 48 hours.
  • KPL cells (Institute of Cell Science, Chinese Academy of Sciences) were cultured with RPMI 1640 (Shanghai Yuanpei, classification number: L210KJ, batch number: F210916) + 10% FBS (Gibco, classification number: 10099-141c, batch number: 2158737cp), and passaged twice.
  • four groups were set up. The first group was the negative control group and culture medium was added. The second group was IN10018 at a concentration of 10 ⁇ M. The third group was Almonertinib at a concentration of 4.7 ⁇ M. The fourth group was IN10018 and Almonertinib.
  • the combination group is IN10018 (10 ⁇ M) and Almonertinib (4.7 ⁇ M). Mix the drugs and incubate in a 5% CO2 incubator at 37°C for 48 hours.
  • KPL cells Institute of Cell Science, Chinese Academy of Sciences
  • RPMI 1640 Session Infection, Chinese Cell Science, Chinese Academy of Sciences
  • FBS FBS
  • Gibco classification number: 10099-141c, batch number: 2158737cp
  • the first group was the negative control group and culture medium was added, and the second group was IN10018.
  • the concentration is 10 ⁇ M
  • the third group is Alflutinib
  • the concentration is 4.7 ⁇ M
  • the fourth group is the combination group of IN10018 and Alflutinib, which are IN10018 (10 ⁇ M) and Alflutinib (4.7 ⁇ M) respectively.
  • Example 4 Study on FAK target inhibition to enhance immunogenic cell death of non-small cell lung cancer cells against AZD9291
  • Anti-calreticulin Recombinant Alexa Fluorescent Anti-Calreticulin (anti-calreticulin) antibody (Abcam, ab196159), Annexin V apoptosis detection kit (Invitrogen, A35110).
  • FAK siRNA is provided by Jima Gene, and the sequence is shown in the table below:
  • HCC827 cells were transfected with control siRNA or FAK siRNA at a final concentration of 50nM respectively. After 24 hours of transfection, 0.3nM AZD9291 was treated for 48 hours, and the cells were stained using Annexin V kit and detected using flow cytometry. The early and late cell apoptosis values were counted and compared with the DMSO control group and plotted using Graphpad 8.0.
  • HCC827 cells were transfected with control siRNA or FAK siRNA at a final concentration of 50nM respectively. 24 hours after transfection, 0.3nM AZD9291 was treated for 48 hours, fluorescent staining was performed with Calreticulin antibody, and the staining results were analyzed by flow cytometry. FlowJo software result statistics show that after FAK silencing combined with AZD9291, the release and exposure of Calreticulin were significantly enhanced, as shown in Figure 8.
  • Example 5 Study on IN10018 enhancing the immunogenic cell death of non-small cell lung cancer cells against AZD9291
  • IN10018 was used in combination with AZD9291 for 6 hours to detect the activation of the endoplasmic reticulum stress signaling pathway; the two drugs were used in combination for 48 hours to detect the expression of Calreticulin, the main target of ICD.
  • Anti-Calreticulin antibody (Abcam, ab196159), FAK antibody (CST, 3285S), Phospho-eIF2 ⁇ (Ser51) (CST, 3398), eif2 ⁇ (CST, 5324), DDIT3 (HUABIO, ET1703-05), HRP-labeled Alpha tubulin ( ⁇ -tubulin) Antibody (Proteintech, HRP-66031), Annexin V apoptosis detection kit (Invitrogen, A35110)
  • a 96-well cell plate was laid with 5000 HCC827 cells/well, and 24 hours later a fixed concentration of IN10018 combined with different concentrations of AZD9291 was added.
  • the concentrations of IN10018 are 3 ⁇ M and 5 ⁇ M; the highest concentration of AZD9291 is 1 ⁇ M, and a total of 9 concentrations are set by 3-fold concentration gradient dilution.
  • 10 ⁇ M CCK8 solution was added to each well.
  • a microplate reader was used to set the absorbance wavelength to 450 nm for reading. The values read were compared to the DMSO control group and plotted using Graphpad 8.0.
  • the results show that AZD9291 at different doses combined with 3 ⁇ M IN10018 and 5 ⁇ M IN10018 for 48 hours has a synergistic killing effect on non-small cell lung cancer cells HCC827, as shown in Figure 9.
  • HCC827 cells were treated with 0.3nM AZD9291 and 3 ⁇ M IN10018. After 48 hours, the cells were collected and stained using Annexin V kit and detected by flow cytometry. The early and late cell apoptosis values were counted and compared with the DMSO control group and plotted using Graphpad 8.0. The results showed that compared with the single drug group, early and late apoptosis were significantly enhanced in the combination group, as shown in Figure 10.
  • HCC827 cells were treated with 0.1nM and 0.3nM AZD9291 and 3 ⁇ M IN10018 respectively, and the protein level expression of endoplasmic reticulum stress-related proteins Phospho-eIF2 ⁇ (Ser51) and DDIT3 was detected 6 hours later.
  • the results showed that compared with the single drug group, the expression levels of Phospho-eIF2 ⁇ (Ser51) and DDIT3 were significantly increased in the combination group, indicating that the endoplasmic reticulum pressure was significantly increased in the combined use of the two drugs, as shown in Figure 11.
  • HCC827 cells were treated with 0.3nM AZD9291 and 3 ⁇ M IN10018. After 48 hours, calreticulin antibody was used for fluorescent staining, and the staining results were analyzed by flow cytometry. FlowJo Software result statistics show that compared with the single drug group, the release and exposure of Calreticulin were significantly enhanced, as shown in Figure 12.
  • Example 6 Study on the in vivo anti-tumor efficacy of AZD9291 in colon cancer CT26 cell BALB/c mouse subcutaneous allograft tumor model
  • mice Female BALB/c mice aged 6-8 weeks were purchased from Shanghai Lingchang Biotechnology Co., Ltd. The experiment began after the animals arrived and were acclimated to the experimental environment. Animals were kept in IVC (independent ventilation system) cages (5 animals per cage) in SPF-level animal rooms. All cages, bedding and drinking water must be sterilized before use. All experimenters should wear protective clothing and latex gloves when operating in the animal room. The animal information card for each cage should indicate the number of animals in the cage, gender, strain, date of receipt, dosage regimen, experiment number, group and experiment start date. Cages, feed and drinking water were changed twice a week. The feeding environment and lighting conditions are as follows:
  • Light cycle 12 hours of light, 12 hours of no light
  • Cage Made of polycarbonate, volume 300mm ⁇ 180mm ⁇ 150mm.
  • the bedding material is corn cobs and is changed twice a week.
  • Drinking water Experimental animals can drink sterilized water freely.
  • Animal identification Experimental animals are identified with ear tags.
  • Colorectal cancer cell CT26 (sourced from Nanjing Kebai Biotechnology Co., Ltd., product number: CBP60043) was maintained and passaged by Yingshi Biotechnology (Nanjing) Co., Ltd.
  • the cells were cultured in a monolayer in vitro.
  • the culture conditions were RPMI-1640 medium plus 10% fetal calf serum and cultured in a 37°C 5% CO 2 incubator. Perform routine digestion with trypsin-EDTA for passage two to three times a week. When the cells are in the exponential growth phase and the saturation is 80%-90%, collect the cells, count them and inoculate them.
  • Administration volume Based on the mouse body weight of 10 mL/kg, if the body weight drops by more than 15%, the animal will stop administration; wait until the body weight recovers to a 10% decrease before resuming administration.
  • the animal's health condition continues to deteriorate, or the tumor volume exceeds 3,000mm 3 , or it has severe disease or pain, it must be euthanized. If the following conditions occur, the veterinarian will be notified and euthanasia will be implemented: obvious weight loss, weight loss of more than 20%; unable to freely eat and drink; the average tumor volume in the control group reaches 2,000mm 3 , and the experiment will be terminated.
  • the animals showed the following clinical manifestations and continued to worsen: piloerection, arched back, white ears, nose, eyes or feet, rapid breathing, convulsions, continuous diarrhea, dehydration, slow movement and vocalization.
  • Tumor diameter was measured with vernier calipers three times a week.
  • the tumor growth inhibition rate TGI (%) was calculated according to the following formula.
  • TGI (%) [1-(Average tumor volume of a certain administration group-Average tumor volume of the administration group at the beginning of treatment)/(Average tumor volume of the solvent control group-Average tumor volume of the solvent control group at the beginning of treatment) area)] ⁇ 100%.
  • the tumor volume in the control group was 2777.3 ⁇ 705.2mm 3 .
  • the tumor volumes of each treatment group of AZD9291+IN10018 (20+25mg/kg), AZD9291+PDL1 antibody (20+10mg/kg) and AZD9291+IN10018+PDL1 antibody (20+25+10mg/kg) were 1693.4 ⁇ 1207.5mm respectively. 3 , 1387.6 ⁇ 859.6mm 3 and 927.2 ⁇ 627.9mm 3 , see Table 4 for details.
  • the tumor rates TGI were 40.0% (p ⁇ 0.0001), 51.2% (p ⁇ 0.0001) and 62.7% (p ⁇ 0.0001) respectively. See Table 4 for details.
  • the comprehensive tumor volume was compared with the AZD9291+IN10018+PDL1 antibody (20+25+10mg/kg) three-drug combination group, and statistical analysis was performed.
  • the tumor volumes of each dose group at different time periods are shown in Figure 13.
  • Table 4 Evaluation of the anti-tumor effect of the test substance on the BALB/c mouse transplant tumor model of mouse colon cancer CT26 cells (based on the data on the 15th day after group administration)
  • the experiment was conducted in accordance with the dosage regimen. During the experiment, the animals were observed for eating, drinking and other activities every day, and the animal weight was recorded three times a week. The animal weight curve is shown in Figure 14. Throughout the entire dosing cycle, the animals in each group showed no significant weight loss and were in good condition.
  • AZD9291+IN10018 (20+25mg/kg), AZD9291+PDL1 antibody (20+10mg/kg) and AZD9291+IN10018+PDL1 antibody (20+25+10mg/kg) treatment groups all had significant The tumor growth inhibition effect was statistically different compared with the control group. Taking into account the entire dosing cycle, the three-drug combination group of AZD9291+IN10018+PDL1 antibody (20+25+10mg/kg) is compared with AZD9291+IN10018 (20+25mg/kg) and AZD9291+PDL1 antibody (20+10mg/kg).
  • the tumor volume in the two-drug combination group has always been smaller, and it is statistically different from the AZD9291+IN10018 (20+25mg/kg) group.
  • the combination of PDL1 antibody (20+10mg/kg) and two drugs has a better effect on inhibiting tumor growth.
  • the animals' weight changed well, and no abnormalities were found in their activities, drinking, eating, and mental status during the entire dosing cycle, indicating that the animals were tolerant to the three-drug combination of AZD9291+IN10018+PDL1 antibody (20+25+10mg/kg).
  • Example 7 Study on the in vivo anti-tumor efficacy of AZD9291 in colon cancer MC38 cell C57BL/6 mouse subcutaneous allograft tumor model
  • mice Female C57BL/6 mice aged 6-8 weeks were purchased from Shanghai Slack Experimental Animal Co., Ltd. The experiment began after the animals arrived and were acclimated to the experimental environment. Animals were kept in IVC (independent ventilation system) cages (5 animals per cage) in SPF-level animal rooms. All cages, bedding and drinking water must be sterilized before use. All experimenters should wear protective clothing and latex gloves when operating in the animal room. The animal information card for each cage should indicate the number of animals in the cage, gender, strain, date of receipt, dosage regimen, experiment number, group and experiment start date. Cages, feed and drinking water are changed twice a week. The feeding environment and lighting conditions are as follows:
  • Light cycle 12 hours of light, 12 hours of no light
  • Cage Made of polycarbonate, volume 300mm ⁇ 180mm ⁇ 150mm.
  • the bedding material is corn cobs and is changed twice a week.
  • Drinking water Experimental animals can drink sterilized water freely.
  • Animal identification Experimental animals are identified with ear tags.
  • Colorectal cancer cell MC38 (sourced from Nanjing Kebai Biotechnology Co., Ltd., product number: CBP60825) was maintained and passaged by Yingshi Biotechnology (Nanjing) Co., Ltd.
  • the cells were cultured in a monolayer in vitro.
  • the culture conditions were DMEM medium plus 10% fetal bovine serum and cultured in a 37°C 5% CO 2 incubator. Perform routine digestion with trypsin-EDTA for passage two to three times a week. When the cells are in the exponential growth phase and the saturation is 80%-90%, collect the cells, count them and inoculate them.
  • Administration volume Based on the mouse body weight of 10 mL/kg, if the body weight drops by more than 15%, the animal will stop administration; wait until the body weight recovers to a 10% decrease before resuming administration.
  • the animal's health condition continues to deteriorate, or the tumor volume exceeds 3,000mm 3 , or it has severe disease or pain, it must be euthanized. If the following conditions occur, the veterinarian will be notified and euthanasia will be implemented: obvious weight loss, weight loss of more than 20%; unable to freely eat and drink; the average tumor volume in the control group reaches 2,000mm 3 , and the experiment will be terminated.
  • the animals showed the following clinical manifestations and continued to worsen: piloerection, arched back, white ears, nose, eyes or feet, rapid breathing, convulsions, continuous diarrhea, dehydration, slow movement and vocalization.
  • Tumor diameter was measured with vernier calipers three times a week.
  • TGI (%) [1-(Average tumor volume of a certain administration group-Average tumor volume of the administration group at the beginning of treatment)/(Average tumor volume of the solvent control group-Average tumor volume of the solvent control group at the beginning of treatment)] ⁇ 100%.
  • the tumor volume in the control group was 1984.4 ⁇ 537.8mm 3 .
  • PD-L1 Tumors in each treatment group of antibody (5mg/kg), AZD9291 (20mg/kg), IN10018+PD-L1 antibody (25+5mg/kg) and AZD9291+IN10018+PD-L1 antibody (20+25+5mg/kg)
  • the volumes are 1974.8 ⁇ 1629.9mm 3 , 1518.1 ⁇ 728.8mm 3 , 1425.2 ⁇ 889.6mm 3 and 1292.9 ⁇ 934.6mm 3 respectively. See Table 8 for details.
  • the comprehensive tumor volume was compared with the AZD9291+IN10018+PD-L1 antibody (20+25+5mg/kg) combination group for statistical analysis.
  • the tumor volumes of each dose group at different time periods are shown in Figure 15.
  • Table 8 Evaluation of the anti-tumor effect of the test substance on the C57BL/6 mouse transplant tumor model of mouse colon cancer MC38 cells (based on the data on the 14th day after group administration)
  • the experiment was conducted according to the dosing plan. During the experiment, the animals were observed for eating, drinking and other activities every day, and the animal weight was recorded three times a week. The animal weight curve is shown in Figure 16.
  • One mouse in the control group was found dead on the 11th day after group administration due to tumor rupture; one mouse in the AZD9291+IN10018+PD-L1 antibody (20+25+5mg/kg) group was found dead due to tumor rupture. Errors in intragastric administration resulted in The animals died on the 8th day. The animals in the remaining groups showed no significant weight loss and were in good condition during the entire dosing cycle, showing tolerance to different dosing methods.
  • both the IN10018 + PD-L1 antibody (25 + 5 mg/kg) and AZD9291 + IN10018 + PD-L1 antibody (20 + 25 + 5 mg/kg) groups had significant tumor growth inhibition effects. There are statistically significant differences. Taking into account the entire dosing cycle, the tumor volume of the AZD9291+IN10018+PD-L1 antibody (20+25+5mg/kg) group has always been smaller than that of the other treatment groups, and is similar to that of the PD-L1 antibody (5mg/kg) alone. There was a statistical difference between the drug groups.
  • Example 8 Study on the induction of immunogenic cell death targets in mouse breast cancer 4T1 cells by AZD9291 and AMP945 in vitro
  • AZD9291 is provided by Shanghai Chaolan Chemical Technology Center, Lot No.: 202012-02
  • AMP945 is provided by MCE, Lot No.:143253
  • 4T1 cells (from Nanjing Kebai Biotechnology Co., Ltd., Cat. No.: CBP60352) were treated with RPMI 1640 (Shanghai Yuanpei, Cat No.: L210KJ, Lot No.: F210916) + 10% FBS (Gibco, Cat No.: 10099-141c , Lot No.: 2158737cp) culture, the culture conditions were 37°C and 5% CO 2 . Twice a week to three passages for routine digestion with trypsin. When the cells are in the exponential growth phase and the adhesion confluence reaches 80%-90%, the cells are harvested and plated. 4T1 cells were digested with trypsin and collected and counted.
  • the cells were diluted with RPMI1640+10% FBS.
  • the dilution concentration was 50,000 cells per ml, and then plated on a 12-well cell culture plate. Each well was Plate 2 ml of cell suspension or 100,000 cells. After plating, the cells were cultured in a 37°C, 5% CO2 incubator. After the cells were spread for 24 hours, six groups were set up. The first group was the control group and culture medium was added. The second group was AMP945 at a concentration of 3 ⁇ M. The third group was AMP945 at a concentration of 6 ⁇ M. The fourth group was AZD9291 at a concentration of 10 ⁇ M.
  • the fifth group is the combination of AMP945 (3 ⁇ M) and AZD9291 (10 ⁇ M)
  • the sixth group is the combination of AMP945 (6 ⁇ M) and AZD9291 (10 ⁇ M). Mix the drugs and incubate at 37°C in a 5% CO2 incubator for 48 hours.
  • Example 9 Study on the induction of immunogenic cell death target in mouse breast cancer 4T1 cells by Alflutinib and AMP945 in vitro
  • Alflutinib is provided by Shanghai MCE, Lot No.:33805
  • AMP945 is provided by MCE, Lot No.:143253
  • 4T1 cells (from Nanjing Kebai Biotechnology Co., Ltd., Cat. No.: CBP60352) were treated with RPMI 1640 (Shanghai Yuanpei, Cat No.: L210KJ, Lot No.: F210916) + 10% FBS (Gibco, Cat No.: 10099-141c , Lot No.: 2158737cp) culture, the culture conditions were 37°C and 5% CO 2 . Perform routine digestion pass-through with trypsin two to three times a week. When the cells are in the exponential growth phase and the adhesion confluence reaches 80%-90%, the cells are harvested and plated. 4T1 cells were digested with trypsin and collected and counted.
  • the cells were diluted with RPMI1640+10% FBS.
  • the dilution concentration was 50,000 cells per ml, and then plated on a 12-well cell culture plate. Each well was Plate 2 ml of cell suspension or 100,000 cells. After plating, the cells were cultured in a 37°C, 5% CO2 incubator. After the cells were spread for 24 hours, six groups were set up. The first group was the control group and culture medium was added. The second group was AMP945 at a concentration of 3 ⁇ M. The third group was AMP945 at a concentration of 6 ⁇ M. The fourth group was Alflutinib at a concentration of 10 ⁇ M.
  • the fifth group is the combination of AMP945 (3 ⁇ M) and Alflutinib (10 ⁇ M)
  • the sixth group is the combination of AMP945 (6 ⁇ M) and Alflutinib (10 ⁇ M). Mix the drugs and incubate at 37°C in a 5% CO2 incubator for 48 hours.
  • Example 10 Study on the induction of immunogenic cell death target in mouse breast cancer 4T1 cells by Almonertinib and AMP945 in vitro
  • Almonertinib is provided by Shanghai MCE, Lot No.: 65966
  • AMP945 is provided by MCE, Lot No.:143253
  • 4T1 cells (from Nanjing Kebai Biotechnology Co., Ltd., Cat. No.: CBP60352) were treated with RPMI 1640 (Shanghai Yuanpei, Cat No.: L210KJ, Lot No.: F210916) + 10% FBS (Gibco, Cat No.: 10099-141c , Lot No.: 2158737cp) culture, the culture conditions were 37°C and 5% CO 2 . Perform routine digestion pass-through with trypsin two to three times a week. When the cells are in the exponential growth phase and the adhesion confluence reaches 80%-90%, the cells are harvested and plated. 4T1 cells were digested with trypsin and collected and counted.
  • the cells were diluted with RPMI1640+10% FBS.
  • the dilution concentration was 50,000 cells per ml, and then plated on a 12-well cell culture plate. Each well was Plate 2 ml of cell suspension or 100,000 cells. After plating, the cells were cultured in a 37°C, 5% CO2 incubator. After the cells were spread for 24 hours, six groups were set up. The first group was the control group and culture medium was added. The second group was AMP945 at a concentration of 3 ⁇ M. The third group was AMP945 at a concentration of 6 ⁇ M. The fourth group was Almonertinib at a concentration of 2 ⁇ M.
  • the fifth group is the combination of AMP945 (3 ⁇ M) and Almonertinib (2 ⁇ M)
  • the sixth group is the combination of AMP945 (6 ⁇ M) and Almonertinib (2 ⁇ M). Mix the drugs and incubate at 37°C in a 5% CO2 incubator for 48 hours.

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Abstract

FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂联合治疗肿瘤,及FAK抑制剂和表皮生长因子受体酪氨酸激酶抑制剂联合治疗肿瘤。

Description

FAK抑制剂及EGFR-TKI的药物组合及用途
本申请要求于2022年9月5日递交的中国专利申请202211077426.8、于2022年9月23日递交的中国专利申请202211166052.7和于2023年8月8日递交的中国专利申请202310998195.2的优先权，在此全文引用上述中国专利申请公开的内容以作为本申请的一部分。
技术领域
本发明属于药物化学领域。具体地，本发明涉及粘着斑激酶(Focal Adhesion Kinase,FAK)抑制剂与其它药物联用治疗肿瘤。
背景技术
肿瘤是威胁人们健康的第二大杀手。免疫原性细胞死亡(Immunogenic cell death，ICD)是基于细胞程序性死亡的叠加效应。癌细胞接触化疗或者靶向治疗类药物的同时可能激活细胞内部的压力信号，这一类压力信号包含内质网压力(ER stress)和活性氧压力(oxidative stress)。在压力信号的作用下细胞会先尝试修复压力，如果压力造成的损伤超出了修复的能力，细胞会启动程序性死亡过程。在这个过程中往往伴随着一类叫做损伤相关分子(Damage associated molecular patterns，DAMPs)的释放，这些分子包括钙网蛋白(calrectulin)、膜联蛋白A1(annexin A1)、高移动组框蛋白1(High mobility group box 1，HMGB1)等。这类DAMPs会特异性的被机体内抗原呈递细胞(Antigen-presenting cells，APC)上的模式识别受体识别，诱导APC成熟、分化和激活，并逐级呈递给效应T细胞等免疫细胞，从而使免疫细胞产生抗原记忆，再次发现相同来源的肿瘤细胞时免疫细胞就会特异性识别并杀伤肿瘤细胞。ICD启动的新的针对肿瘤的特异性免疫反应可增加对免疫检查点抑制剂(Immuno-checkpoint inhibitor，ICI)的敏感，以此增效免疫检查点类抑制剂的作用，并可产生具有免疫记忆持久的抗肿瘤的反应。
FAK，又称为蛋白酪氨酸激酶2(PTK2)，是一种非受体酪氨酸激酶，并且是粘着斑复合体的关键组分。FAK在介导整合素和生长因子信号以调节肿瘤细胞的侵袭、增殖和存活方面发挥着重要作用。
EGFR是表皮生长因子受体(Epidermal Growth Factor Receptor)的简称。 EGFR基因负责编码并制造一种称为表皮生长因子受体的受体蛋白。EGFR受体蛋白是一种跨膜蛋白，其分为三部分：蛋白的一端位于细胞外，一部分位于细胞膜，另一端则位于细胞内。这允许EGFR受体与细胞外的其他蛋白(称为配体)结合，帮助细胞接收信号并对其刺激作出反应。而受体与配体的结合如同钥匙与锁，因此它们都有特定的结合“伙伴”。当EGFR与配体结合时，它会附着于另一个位于附近的EGFR受体并形成复合物(二聚体)，从而进入激活状态，并激活细胞内的信号传导途径。EGFR突变主要发生在18～21号外显子，其中19号外显子的缺失突变和21号外显子的L858R点突变是最常见的突变类型，占所有突变类型的90％。当EGFR发生致病性基因突变时，EGFR受体蛋白便会处于持续激活状态，这导致细胞持续接收增生和生存的信号，造成细胞过度生长及存活(不能正常凋亡)，导致肿瘤的形成。
目前上市针对EGFR突变的靶向药物包括：一代针对19、21外显子突变的埃克替尼(Icotinib)、吉非替尼(gefitinib)、厄洛替尼(erlotinib)；二代针对8、20号外显子突变的阿法替尼(afatinib)及三代针对T790M突变的奥希替尼(osimertinib，在本文中也称作AZD9291)、阿美替尼(Almonertinib)及艾氟替尼(Alflutinib)。针对ALK突变的靶向药物包括：一代靶向药物克唑替尼(crizotinib)，二代靶向药色瑞替尼(ceritinib)、艾乐替尼(alectinib)、布格替尼(brigatinib)及三代的靶向药物劳拉替尼(lorlatinib)等。但这些靶向药物耐药性多出现于用药后1年左右。克服靶向药物的耐药性，或推迟耐药时间及提高治愈的可能是药物研发的主要目标。
因此，仍然需要找到一种方法去提高靶向中单药的疗效，提高应答率，通过联合用药为免疫治疗创造条件，并进一步克服耐药，为肿瘤治愈提供可能。
发明内容
本公开一方面提供了FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂在制备用于在对象中治疗肿瘤的药物中的用途。
本公开又一方面提供了FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂的药物组合产品，其用于在对象中治疗肿瘤。
本公开又一方面提供了一种治疗肿瘤的方法，该方法包括向有需要的对 象施用治疗有效量的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂。
本公开又一方面提供了一种试剂盒或药学上可接受的组合物，其包括：(a)FAK抑制剂；(b)表皮生长因子受体酪氨酸激酶抑制剂；和(c)免疫检查点抑制剂。
本公开又一方面提供了FAK抑制剂和表皮生长因子受体酪氨酸激酶抑制剂在制备用于在治疗肿瘤的药物中的用途，其中所述FAK抑制剂用于增强所述表皮生长因子受体酪氨酸激酶抑制剂诱导的免疫原性细胞死亡。
本公开又一方面提供了FAK抑制剂，其在治疗肿瘤中用于增强表皮生长因子受体酪氨酸激酶抑制剂诱导的免疫原性细胞死亡。
本公开又一方面提供了一种治疗肿瘤的方法，该方法包括向有需要的对象施用治疗有效量的FAK抑制剂和表皮生长因子受体酪氨酸激酶抑制剂，其中所述FAK抑制剂用于增强所述表皮生长因子受体酪氨酸激酶抑制剂诱导的免疫原性细胞死亡。
本公开又一方面提供了FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂在制备用于联合治疗肿瘤的药物中的用途。
本公开又一方面提供了FAK抑制剂在制备用于与表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂治疗肿瘤的联用药物中的用途。
本公开又一方面提供了表皮生长因子受体酪氨酸激酶抑制剂在制备用于与FAK抑制剂和免疫检查点抑制剂治疗肿瘤的联用药物中的用途。
本公开又一方面提供了免疫检查点抑制剂在制备用于与FAK抑制剂和表皮生长因子受体酪氨酸激酶抑制剂治疗肿瘤的联用药物中的用途。
本公开又一方面提供了FAK抑制剂在制备用于与表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂联合治疗肿瘤的药物中的用途。
本公开又一方面提供了表皮生长因子受体酪氨酸激酶抑制剂在制备用于与FAK抑制剂和免疫检查点抑制剂联合治疗肿瘤的药物中的用途。
本公开又一方面提供了免疫检查点抑制剂在制备用于与FAK抑制剂和表皮生长因子受体酪氨酸激酶抑制剂联合治疗肿瘤的药物中的用途。
本公开又一方面提供了一种试剂盒，其包括：FAK抑制剂；和说明书，该说明书指出该FAK抑制剂可用于与表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂联合治疗肿瘤。
本公开又一方面提供了一种试剂盒，其包括：表皮生长因子受体酪氨酸激酶抑制剂；和说明书，该说明书指出该表皮生长因子受体酪氨酸激酶抑制剂可用于与FAK抑制剂和免疫检查点抑制剂联合治疗肿瘤。
本公开又一方面提供了一种试剂盒，其包括：免疫检查点抑制剂；和说明书，该说明书指出该免疫检查点抑制剂可用于与FAK抑制剂和表皮生长因子受体酪氨酸激酶抑制剂联合治疗肿瘤。
本公开另一方面提供了一种治疗肿瘤的方法，该方法包括向有需要的对象施用治疗有效量的FAK抑制剂和表皮生长因子受体酪氨酸激酶抑制剂。
本公开又一方面提供了FAK抑制剂和表皮生长因子受体酪氨酸激酶抑制剂的药物组合产品，其用于在有需要的对象中治疗肿瘤。
本公开又一方面提供了FAK抑制剂和表皮生长因子受体酪氨酸激酶抑制剂在制备用于治疗肿瘤的联用药物中的用途。
本公开又一方面提供了FAK抑制剂在制备用于与表皮生长因子受体酪氨酸激酶抑制剂治疗肿瘤的联用药物中的用途。
本公开又一方面提供了表皮生长因子受体酪氨酸激酶抑制剂在制备用于与FAK抑制剂治疗肿瘤的联用药物中的用途。
本公开又一方面提供了FAK抑制剂和表皮生长因子受体酪氨酸激酶抑制剂在制备用于联合治疗肿瘤的药物中的用途。
本公开又一方面提供了FAK抑制剂在制备用于与表皮生长因子受体酪氨酸激酶抑制剂联合治疗肿瘤的药物中的用途。
本公开又一方面提供了表皮生长因子受体酪氨酸激酶抑制剂在制备用于与FAK抑制剂联合治疗肿瘤的药物中的用途。
本公开又一方面提供了一种试剂盒，其包括：FAK抑制剂；和说明书，该说明书指出该FAK抑制剂可用于与表皮生长因子受体酪氨酸激酶抑制剂联合治疗肿瘤。
本公开又一方面提供了一种试剂盒，其包括：表皮生长因子受体酪氨酸激酶抑制剂；和说明书，该说明书指出该表皮生长因子受体酪氨酸激酶抑制剂可用于与FAK抑制剂联合治疗肿瘤。
可选的，所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、APG-2449、AMP945、AMP886或其药学上可接受的盐，优选为IN10018、AMP945、Defactinib、或其药学上可接受的盐，进 一步优选为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐，所述IN10018结构如下：
所述Defactinib也称为地法替尼，CAS号为1345713-71-4；所述GSK2256098的CAS号为1224887-10-8。所述PF-00562271的CAS号为717907-75-0；所述VS-4718的CAS号为1061353-68-1；所述APG-2449为亚盛医药研发；所述AMP945的CAS号为1393653-34-3。
可选的，所述表皮生长因子受体酪氨酸激酶抑制剂为吉非替尼(Gefitinib)、厄洛替尼(Erlotinib)、埃克替尼(Icotinib)、阿法替尼(Afatinib)、达克替尼(Dacomitinib)、克唑替尼(Crizotinib)、Osimertinib(奥希替尼，AZD9291)、阿美替尼(Almonertinib)、艾氟替尼(Alflutinib，又名伏美替尼(Furmonertinib))、EAI045、JBJ-04-125-02、BLU-945、BLU-701、TQB3804、BBT-176、ES-072、BPI-361175、CH7233163或其药学上可接受的盐。
所述吉非替尼(Gefitinib)的CAS号为184475-35-2；厄洛替尼(Erlotinib)的CAS号为183321-74-6；埃克替尼(Icotinib)的CAS号为610798-31-7；阿法替尼(Afatinib)的CAS号为850140-72-6；克唑替尼(Crizotinib)的CAS号为877399-52-5；Osimertinib(奥希替尼，AZD9291)的CAS号为1421373-65-0；阿美替尼(Almonertinib)的CAS号为1899921-05-1；艾氟替尼(Alflutinib，又名伏美替尼(Furmonertinib))的CAS号为1869057-83-9；EAI045的CAS号为1942114-09-1；JBJ-04-125-02的CAS号为2060610-53-7；BLU-945的CAS号为2660250-10-0；BLU-701是缆图药业公司(Blueprint Medicines Corp)和再鼎医药联合开发的；TQB3804的CAS号为2267329-76-8；BBT-176是毕利吉生物科技股份有限公司开发的；ES-072是博生医药开发的；BPI-361175是贝达药业研发的；CH7233163是中外制药株式会社开发的。
可选的，所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib、Almonertinib、Alflutinib或其药学上可接受的盐。
可选的，所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib或其药 学上可接受的盐。
可选的，所述表皮生长因子受体酪氨酸激酶抑制剂为Almonertinib或其药学上可接受的盐。
可选的，所述表皮生长因子受体酪氨酸激酶抑制剂为Alflutinib或其药学上可接受的盐。
可选的，所述免疫检查点抑制剂为抗PD-1/PD-L1抗体、PD-1/PD-L1小分子抑制剂或者TIGIT抑制剂。
可选的，所述免疫检查点抑制剂为抗PD-1/PD-L1抗体，进一步的，所述抗PD-1/PD-L1抗体为帕博利珠单抗(pembrolizumab)、替雷利珠单抗(Tislelizumab)、尼伏单抗(Nivolumab)、特瑞普利单抗(Toripalimab)、阿替利珠单抗(Atezolizumab)、度伐单抗(durvalumab)、阿维单抗(Avelumab)、卡瑞利珠单抗(Camrelizumab)、信迪利单抗(Sintilimab)、西米普利单抗(Cemiplimab)、恩沃利单抗(envafolimab)、BMS-936559、JS003、SHR-1316、GS-4224、AN-4005或者MX-10181。
可选的，所述免疫检查点抑制剂为PD-1/PD-L1小分子抑制剂，进一步的，所述PD-1/PD-L1小分子抑制剂为INCB-086550、拉泽替尼(Lazertinib)、IMMH-010、CA-170、ABSK043或者RRx-001。
可选的，所述免疫检查点抑制剂为TIGIT抑制剂，进一步的，所述TIGIT抑制剂为欧司珀利单抗(Ociperlimab/BGB-A1217)、维博利单抗(Vibostolimab)、domvanalimab(AB154)、替瑞利尤单抗(Tiragolumab)、Belrestotug、艾替利单抗(Etigilimab)、ONO-4686、JS-006、AZD-2936、HLX-301、SEA-TGT、M-6223、IBI-939、COM-902、AB-308、AGEN-1777、AK-127、BAT-6021、BAT-6005、ASP-8374、PM-1022、BMS-986207、HB0036或IBI-321。
可选的，所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂。
可选的，所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体。
可选的，所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表 皮生长因子受体酪氨酸激酶抑制剂为Almonertinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂。
可选的，所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Almonertinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体。
可选的，所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Alflutinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂。
可选的，所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Alflutinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体。
可选的，所述FAK抑制剂、所述表皮生长因子受体酪氨酸激酶抑制剂和所述免疫检查点抑制剂被同时或依次施用于所述对象。
可选的，所述肿瘤为膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、神经胶质瘤胡或恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)。
可选的，所述肿瘤优选为乳腺癌、卵巢癌、结肠癌(包括结直肠癌)、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤或胰腺癌。
可选的，所述肿瘤为肺癌(包括小细胞肺癌和非小细胞肺癌)或结肠癌(包括结直肠癌)。
可选的，所述肿瘤为非小细胞肺癌或结肠癌(包括结直肠癌)。
附图说明
为了更清楚地说明本公开实施例的技术方案，下面将对实施例的附图作简单地介绍，显而易见地，下面描述中的附图仅仅涉及本公开的一些实施例，而非对本发明的限制。
图1显示了实施例1肺癌KPL细胞与药物孵育48小时后，拍摄细胞的白光显微镜照片。
图2a显示了实施例1肺癌KPL细胞与药物孵育48小时后，CRT阳性KPL细胞的百分比；图2b显示了实施例1肺癌KPL细胞与药物孵育48小时后，Annexin V阳性KPL细胞的百分比。
图3显示了实施例2肺癌KPL细胞与药物孵育48小时后，拍摄细胞的白光显微镜照片。
图4a显示了实施例2肺癌KPL细胞与药物孵育48小时后，CRT阳性KPL细胞的百分比；图4b显示了实施例2肺癌KPL细胞与药物孵育48小时后，Annexin V阳性KPL细胞的百分比。
图5显示了实施例3肺癌KPL细胞与药物孵育48小时后，拍摄细胞的白光显微镜照片。
图6a显示了实施例3肺癌KPL细胞与药物孵育48小时后，CRT阳性KPL细胞的百分比；图6b显示了实施例3肺癌KPL细胞与药物孵育48小时后，Annexin V阳性KPL细胞的百分比。
图7显示了FAK沉默后联合0.3nM AZD9291处理48小时后细胞早期及晚期凋亡情况。
图8显示了FAK沉默后联合0.3nM AZD9291处理48小时后增强了Calreticulin释放及暴露。
图9显示了不同剂量下AZD9291分别与3μM IN10018和5μM IN10018合用48小时对非小细胞肺癌细胞HCC827的杀伤协同作用。
图10显示了0.3nM AZD9291与3μM IN10018合用48小时后检测细胞凋亡。
图11显示了AZD9291与IN10018合用显著上调了HCC827的内质网应激。
图12显示了AZD9291与IN10018合用增强了Calreticulin释放及暴露。
图13显示了在结肠癌CT26细胞BALB/c小鼠皮下同种移植瘤模型中给予不同受试物后的肿瘤体积的变化。
图14显示了在结肠癌CT26细胞BALB/c小鼠皮下同种移植瘤模型中给予不同受试物后小鼠体重的变化。
图15显示了在结肠癌MC38细胞C57BL/6小鼠皮下同种移植瘤模型中给予不同受试物后的肿瘤体积的变化。
图16显示了在结肠癌MC38细胞C57BL/6小鼠皮下同种移植瘤模型中给予不同受试物后小鼠体重的变化。
图17a显示了实施例8乳腺癌4T1细胞与药物孵育48小时后，CRT阳性4T1细胞的百分比；图17b显示了实施例8乳腺癌4T1与药物孵育48小时后，Annexin V阳性4T1细胞的百分比。
图18a显示了实施例9乳腺癌4T1细胞与药物孵育48小时后，CRT阳性4T1细胞的百分比；图18b显示了实施例9乳腺癌4T1与药物孵育48小时后，Annexin V阳性4T1细胞的百分比。
图19a显示了实施例10乳腺癌4T1细胞与药物孵育48小时后，CRT阳性4T1细胞的百分比；图19b显示了实施例10乳腺癌4T1与药物孵育48小时后，Annexin V阳性4T1细胞的百分比。
具体实施方式
为使本公开实施例的目的、技术方案和优点更加清楚，下面将结合本公开实施例的附图，对本公开实施例的技术方案进行清楚、完整地描述。显然，所描述的实施例是本公开的一部分实施例，而不是全部的实施例。基于所描述的本公开的实施例，本领域普通技术人员在无需创造性劳动的前提下所获得的所有其他实施例，都属于本发明保护的范围。
本发明可在不偏离本发明基本属性的情况下以其它具体形式来实施。应该理解的是，在不冲突的前提下，本发明的任一和所有实施方案都可与任一其它实施方案或多个其它实施方案中的技术特征进行组合以得到另外的实施方案。本发明包括这样的组合得到的另外的实施方案。
本公开中提及的所有出版物和专利在此通过引用以它们的全部内容纳入本公开。如果通过引用纳入的任何出版物和专利中使用的用途或术语与本公开中使用的用途或术语冲突，那么以本公开的用途和术语为准。
本文所用的章节标题仅用于组织文章的目的，而不应被解释为对所述主题的限制。
除非另有规定，本文使用的所有技术术语和科学术语具有要求保护主题所属领域的通常含义。倘若对于某术语存在多个定义，则以本文定义为准。
除了在工作实施例中或另外指出之外，在说明书和权利要求中陈述的定量性质例如剂量的所有数字应理解为在所有情况中被术语“约”修饰。还应理解的是，本申请列举的任何数字范围意在包括该范围内的所有的子范围和该范围或子范围的各个端点的任何组合。
本公开中使用的“包括”、“含有”或者“包含”等类似的词语意指出现该词前面的要素涵盖出现在该词后面列举的要素及其等同，而不排除未记载的要素。本文所用的术语“含有”或“包括(包含)”可以是开放式、半封闭式和封闭式的。换言之，所述术语也包括“基本上由…组成”、或“由…组成”。
定义
本申请中所用的下列术语和符号具有如下所述的含义，其所处的上下文中另有说明的除外。本文所用的术语“FAK抑制剂”是指FAK的有效抑制剂，可适于哺乳动物，特别是人。在一些实施方案中，所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、APG-2449、AMP945、AMP886或其药学上可接受的盐，所述IN10018结构如下：
所述Defactinib也称为地法替尼，CAS号为1345713-71-4；所述GSK2256098的CAS号为1224887-10-8。所述PF-00562271的CAS号为717907-75-0；所述VS-4718的CAS号为1061353-68-1；所述APG-2449为亚盛医药研发；所述AMP945的CAS号为1393653-34-3。
在一些实施方案中，所述FAK抑制剂优选为IN10018、AMP945、Defactinib或其药学上可接受的盐，在一些优选实施方案中，所述FAK抑制剂为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐。
本文所用术语“表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKI)”是可选择性地、有效地抑制表皮生长因子受体酪氨酸激酶的药剂。所述表皮生长因子受体酪氨酸激酶抑制剂的实例包括但不限于吉非替尼(Gefitinib)、厄洛替 尼(Erlotinib)、埃克替尼(Icotinib)、阿法替尼(Afatinib)、达克替尼(Dacomitinib)、克唑替尼(Crizotinib)、Osimertinib(奥希替尼，AZD9291)、阿美替尼(Almonertinib)、艾氟替尼(Alflutinib，又名伏美替尼(Furmonertinib))、EAI045、JBJ-04-125-02、BLU-945、BLU-701、TQB3804、BBT-176、ES-072、BPI-361175、CH7233163或其药学上可接受的盐。在一些实施方案中，所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib、Almonertinib、Alflutinib或其药学上可接受的盐。
所述吉非替尼(Gefitinib)的CAS号为184475-35-2；厄洛替尼(Erlotinib)的CAS号为183321-74-6；埃克替尼(Icotinib)的CAS号为610798-31-7；阿法替尼(Afatinib)的CAS号为850140-72-6；达克替尼(Dacomitinib)的CAS号为1110813-31-4；克唑替尼(Crizotinib)的CAS号为877399-52-5；Osimertinib(奥希替尼，AZD9291)的CAS号为1421373-65-0；阿美替尼(Almonertinib)的CAS号为1899921-05-1；艾氟替尼(Alflutinib，又名伏美替尼(Furmonertinib))的CAS号为1869057-83-9；EAI045的CAS号为1942114-09-1；JBJ-04-125-02的CAS号为2060610-53-7；BLU-945的CAS号为2660250-10-0；BLU-701是缆图药业公司(Blueprint Medicines Corp)和再鼎医药联合开发的；TQB3804的CAS号为2267329-76-8；BBT-176是毕利吉生物科技股份有限公司开发的；ES-072是博生医药开发的；BPI-361175是贝达药业研发的；CH7233163是中外制药株式会社开发的。
本文所用的术语“免疫检查点抑制剂”是指能够通过调控免疫检查点通路(例如PD-1、TIGIT、CTLA-4、LAG-3、TIM-3等)来提高免疫系统活性的药物。在一些实施方案中，免疫检查点抑制剂为PD-1/PD-L1(程序性细胞死亡蛋白1)通路的拮抗剂(也称为“PD-1抑制剂”)或TIGIT抑制剂。PD-1抑制剂在本公开中也称为PD-1/PD-L1抑制剂。例如，在本公开的治疗方法、药物和用途中所述PD-1/PD-L1抑制剂为PD-1/PD-L1抗体，包括但不限于帕博利珠单抗(可瑞达/Keytruda)、替雷利珠单抗(Tislelizumab/百泽安)、尼伏单抗(Nivolumab)、特瑞普利单抗(Toripalimab/拓益)、度伐单抗(英飞凡/度伐鲁单抗/durvalumab)、阿维单抗(Avelumab/Bavencio)、阿替利珠单抗(MPDL3280A/Atezolizumab/Tecentriq)、BMS-936559(全人源抗PD-L1的IgG4单克隆抗体)、GS-4224、AN-4005或者MX-10181。在一些优选的实施方案中，PD-1抑制剂为特瑞普利单抗。在一些实施方案中，PD-1抑制剂用于治 疗人对象。在一些实施方案中，PD-1为人PD-1。PD-1/PD-L1抑制剂也包括PD-1/PD-L1小分子抑制剂，例如INCB-086550、拉泽替尼(Lazertinib)、IMMH-010、CA-170、ABSK043或者RRx-001。TIGIT(也称为WUCAM、Vstm3、VSIG9)是Ig超家族的一种受体，是继PD-1/PD-L1之后的新型免疫检查点。例如，在本公开的治疗方法、药物和用途中，所述免疫检查点抑制剂为TIGIT抑制剂，包括但不限于欧司珀利单抗(Ociperlimab/BGB-A1217)、维博利单抗(Vibostolimab)、domvanalimab(AB154)、替瑞利尤单抗(Tiragolumab)、Belrestotug、艾替利单抗(Etigilimab)、ONO-4686、JS-006、AZD-2936、HLX-301、SEA-TGT、M-6223、IBI-939、COM-902、AB-308、AGEN-1777、AK-127、BAT-6021、BAT-6005、ASP-8374、PM-1022、HB0036或IBI-321。在一些实施方案中，TIGIT抑制剂用于治疗人对象。为了避免歧义，本文中抗体均包括双抗。
本文所用的“药物组合”或者“药物组合产品”既可以指采用一个剂量单位形式的固定组合(例如所有药物活性成分以一种剂型存在)或者成套药盒以组合施用的产品的情形，也可以指一种药物与指示该药物可与另外的一种或多种药物联合使用的说明书的组合情形。
本文所用的“联合治疗”或者“联用药物”是指一种药物与另外的一种或多种药物联合使用来治疗疾病，既包括一种药物与另外的一种或多种药物的组合的情形，也包括一种药物与指示该药物可与另外的一种或多种药物联合使用的说明书的组合情形。
“同时或依次施用”在本申请中是指一个给药周期内(例如4周内、3周内、2周内、1周内或24小时以内)两种以上的药物同时或以一定时间间隔先后施用，药物施用的方式(例如口服、静脉、肌肉或皮下施用等)可以相同或不同，两种以上的药物的给药频率/周期可以相同或不同。当本公开的治疗方法、产品或用途涉及两种药物时，两种药物可同时或以一定时间间隔分别单独施用。当本公开的治疗方法、产品或用途涉及三种药物时，三种药物可以在同一时间点施用，或者两种药物在一个时间点施用而剩下一种药物在另一个时间点施用，或者所有三种药物各自在不同时间点施用。
在一些实施方案中，PD-1/PD-L1抑制剂被静脉(例如，作为静脉输注)或皮下施用或口服。优选地，PD-1/PD-L1抑制剂以静脉输注施用。
在一些实施方案中，TIGIT抑制剂被静脉(例如，作为静脉输注)或皮下 施用或口服。优选地，TIGIT抑制剂以静脉输注施用。
免疫检查点抑制剂治疗癌症的能力取决于肿瘤组织内肿瘤抗原特异性T细胞的存在。这要求肿瘤组织表达将自身与其非转化的对应物区分开来的抗原，例如，通过被称为新抗原(neoantigen)的新型蛋白产物。肿瘤新抗原负荷与免疫原性和敏感性(例如，对检查点抑制剂疗法的敏感性)密切相关，这意味着免疫原性较差的肿瘤应该在很大程度上对这些药物耐药。用于释放可被APC摄取的肿瘤抗原的疗法，如诱导免疫原性细胞死亡(ICD)的那些，可能会促进有效的抗肿瘤免疫，特别是当进一步与检查点抑制剂联合时。
本文所用的术语“治疗”是指给患有疾病或者具有所述疾病的症状的对象施用一种或多种药物物质，用以治愈、缓解、减轻、改变、医治、改善、改进或影响所述疾病或者所述疾病的症状。在一些实施方案中，所述疾病是肿瘤或者癌症。
本文所用的术语“肿瘤”是指机体在各种致瘤因素的作用下，局部组织的细胞在基因水平上失去对其生长的正常调控，从而导致其克隆型异常增生而形成的异常病变。所述肿瘤包括，但不限于：膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤，神经胶质瘤以及恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)；在一些实施方案中，所述肿瘤优选为乳腺癌、卵巢癌、结肠癌(包括结直肠癌)、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤和胰腺癌；在一些实施方案中，所述肿瘤为肺癌(包括小细胞肺癌和非小细胞肺癌)或结肠癌(包括结直肠癌)；在一些实施方案中，所述肿瘤为非小细胞肺癌或结肠癌(包括结直肠癌)。
本文所用的术语“对象”或“受试者”是指哺乳动物和非哺乳动物。哺乳动物是指哺乳类的任何成员，其包括但不限于：人；非人灵长类动物，如 黑猩猩及其它猿类和猴类物种；农场动物，如牛、马、绵羊、山羊和猪；家畜，如兔、狗和猫；实验室动物，包括啮齿类动物，如大鼠、小鼠和豚鼠；等等。非哺乳动物的例子包括但不限于鸟等。术语“对象”并不限定特定的年龄或性别。在一些实施方案中，对象是人。
本文所用的术语“药学上可接受的”指的是无毒的、生物学上可耐受的，适合给对象施用的。
本文所用的术语“药学上可接受的盐”指的是无毒的、生物学上可耐受的适合给对象施用的酸加成盐，包括但不限于：与无机酸形成的酸加成盐，例如盐酸盐、氢溴酸盐、碳酸盐、碳酸氢盐、磷酸盐、硫酸盐、亚硫酸盐、硝酸盐等；以及与有机酸形成的酸加成盐，例如甲酸盐、乙酸盐、苹果酸盐、马来酸盐、富马酸盐、酒石酸盐、琥珀酸盐、柠檬酸盐、乳酸盐、甲磺酸盐、对甲苯磺酸盐、2-羟基乙磺酸盐、苯甲酸盐、水杨酸盐、硬脂酸盐和与式HOOC-(CH2)n-COOH(其中n是0-4)的链烷二羧酸形成的盐等。
此外，药学上可接受的酸加成盐可以按照由碱性化合物制备酸加成盐的常规操作通过将游离碱溶于合适的溶剂并且用酸处理该溶液来得到。本领域技术人员无需过多实验即可确定各种可用来制备无毒的药学上可接受的酸加成盐的合成方法。
本文所用的术语“药学上可接受的组合物”是指必须在化学和/或毒理学上与包括制剂的其他成分相容，和/或与接受其治疗的对象相容。本文所用的术语“治疗有效量”是指通常足以对对象产生有益治疗效果的量。可以通过常规方法(例如建模、剂量递增研究或临床试验)结合常规影响因素(例如给药方式、化合物的药代动力学、疾病的严重程度和病程、对象的病史、对象的健康状况、对象对药物的响应程度等)来确定本发明的治疗有效量。
本文所用的术语“抑制”是指生物活动或过程的基线活性的降低。
本文所用的术语“试剂盒”是指用于盛放检测化学成分、药物残留、病毒种类等化学试剂的盒子。本发明所述试剂盒可以是包括(i)FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂中的一种、二种或三种；以及(ii)说明书，所述说明书指出可使用FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂来在对象中治疗肿瘤。在一种实施方案中，试剂盒包括(i)FAK抑制剂；以及(ii)说明书，所述说明书指出可使用FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂 来在对象中治疗肿瘤。在一种实施方案中，试剂盒包括(i)表皮生长因子受体酪氨酸激酶抑制剂；以及(ii)说明书，所述说明书指出可使用FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂来在对象中治疗肿瘤。在一种实施方案中，试剂盒包括(i)免疫检查点抑制剂；以及(ii)说明书，所述说明书指出可使用FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂来在对象中治疗肿瘤。在一种实施方案中，试剂盒包括(i)FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂；以及(ii)说明书，所述说明书指出可使用FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂来在对象中治疗肿瘤。在一种实施方案中，试剂盒包括(i)FAK抑制剂；以及(ii)说明书，所述说明书指出可使用FAK抑制剂和表皮生长因子受体酪氨酸激酶抑制剂来在对象中治疗肿瘤。在一种实施方案中，试剂盒包括(i)表皮生长因子受体酪氨酸激酶抑制剂；以及(ii)说明书，所述说明书指出可使用表皮生长因子受体酪氨酸激酶抑制剂和FAK抑制剂在对象中治疗肿瘤。
试剂盒的化合物可以包含在分开的容器中。可选地，两种或更多种化合物包含在同一容器中。例如，试剂盒可以包括第一容器、第二容器、第三容器和包装插页，其中第一容器包括至少一个剂量的包括FAK抑制剂的药物，第二容器包括至少一个剂量的表皮生长因子受体酪氨酸激酶抑制剂，第三容器包括至少一个剂量的免疫检查点抑制剂的药物，且所述包装插页包括使用药物治疗对象的肿瘤的说明。第一容器、第二容器和第三容器可以包含相同或不同形状(例如，小瓶、注射器和瓶)和/或材料(例如，塑料或玻璃)。试剂盒还可以包括可以有助于施用药物的其他材料，如稀释剂、过滤器、IV袋和管线、针和注射器。
给予受试者的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂、或免疫检查点抑制剂的精确量将取决于各种因素，例如给定的药物或化合物，药物制剂，给药途径，疾病类型，病症，所治疗的受试者或宿主的身份等，但是仍然可以由本领域技术人员常规确定。例如，确定有效量还取决于细胞增殖的程度，严重性和类型。技术人员将能够根据这些和其他因素确定合适的剂量。
FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂、或免疫检查点抑制剂可选择合适的方式例如口服、静脉、肌肉或皮下施用给药。
例如，口服给药时，可以将药物与药学上可接受的载体例如惰性稀释剂或可吸收的食用载体一起口服给药。它们可以封装在硬壳或软壳明胶胶囊中，可以压制成片剂，或者可以直接与患者的食物混合。例如，药物可以与一种或多种赋形剂组合，并以可摄取的片剂，口腔片剂，锭剂，胶囊，酏剂，悬浮液，糖浆或糯米纸囊剂等形式使用。片剂，锭剂，丸剂，胶囊剂等可进一步包括：粘合剂，例如黄芪胶，阿拉伯胶，玉米淀粉或明胶；赋形剂，如磷酸二钙；崩解剂，例如玉米淀粉，马铃薯淀粉，海藻酸等；润滑剂，例如硬脂酸镁；或甜味剂，例如蔗糖，果糖，乳糖或阿斯巴甜；或调味剂。
例如，输注或注射静脉内或腹膜内给药时，药物的溶液可以在水中制备，任选地与无毒的表面活性剂混合。
用于注射或输注的示例性药物剂型包括：无菌水溶液，分散液，或包含活性成分的无菌粉末，该无菌粉末适合于临时制备无菌注射或输注溶液或分散液。无论如何，最终剂型在生产和储存条件下均应无菌，流动且稳定。
无菌注射溶液可以通过将所需量的药物与所需的上述各种其他成分掺入适当的溶剂中，然后过滤灭菌来制备。对于用于制备无菌注射溶液的无菌粉末，优选的制备方法可以是真空干燥和冷冻干燥技术，其可以产生活性成分加上先前无菌过滤后存在的任何其他所需成分的粉末。
用于治疗所需的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂、或免疫检查点抑制剂的量不仅可以随所选择的特定试剂而变化，还可以随给药途径，所治疗疾病的性质以及患者的年龄和状况而变化，并且最终可以由主治医师或临床医生自行决定。然而，一般而言，剂量可以在每天约0.1至约50mg/kg体重的范围内。
所述FAK抑制剂以成年人中5mg/天-300mg/天的剂量范围施用。在一种具体的实施方案中，IN10018或其药学上可接受盐以成年人中5mg/天-100mg/天的剂量施用，在一种具体的实施方案中，IN10018或其药学上可接受盐以成年人中25mg/天-100mg/天的剂量施用，所述剂量以游离碱计。
所述表皮生长因子受体酪氨酸激酶抑制剂以成年人中每天2-250mg的剂量范围施用。在一种具体的实施方式中，Osimertinib或其药学上可接受盐以成年人中每天2-250mg，例如80mg的剂量施用，所述剂量Osimertinib计；Almonertinib或其药学上可接受盐以成年人中每天2-250mg，例如110mg的剂量施用，所述剂量以Almonertinib计；Alflutinib或其药学上可接受盐以成 年人中每天2-250mg，例如80mg的剂量施用，所述剂量以Alflutinib计。
所述免疫检查点抑制剂每次给药以成年人中2-10mg/kg或者50-1200mg的剂量施用，每2周到3周给药一次。在一种具体的实施方式中，所述免疫检查点抑制剂每次给药以成年人中3-10mg/kg或者100-1200mg的剂量施用，每2周到3周给药一次。
本文所用的未具体定义的技术和科学术语具有本发明所属领域的技术人员通常理解的含义。
在一些实施方案中，本公开还公开了以下：
1.FAK抑制剂、所述表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其用于在对象中治疗肿瘤的方法。
2.如实施方案1所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、APG-2449、AMP945、AMP886或其药学上可接受的盐，优选为IN10018、AMP945、Defactinib、或其药学上可接受的盐，进一步优选为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐，所述IN10018结构如下：
3.如实施方案1-2所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述表皮生长因子受体酪氨酸激酶抑制剂为吉非替尼(Gefitinib)、厄洛替尼(Erlotinib)、埃克替尼(Icotinib)、阿法替尼(Afatinib)、达克替尼(Dacomitinib)、克唑替尼(Crizotinib)、Osimertinib(奥希替尼，AZD9291)、阿美替尼(Almonertinib)、艾氟替尼(Alflutinib，又名伏美替尼(Furmonertinib))、EAI045、JBJ-04-125-02、BLU-945、BLU-701、TQB3804、BBT-176、ES-072、BPI-361175、CH7233163或其药学上可接受的盐。
4.如实施方案1-3任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib、Almonertinib、Alflutinib或其药学上可接受的盐。
5.如实施方案1-4任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib或其药学上可接受的盐。
6.如实施方案1-4任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Almonertinib或其药学上可接受的盐。
7.如实施方案1-4任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Alflutinib或其药学上可接受的盐。
8.如实施方案1-7任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体、PD-1/PD-L1小分子抑制剂或者TIGIT抑制剂。
9.如实施方案1-8任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体，进一步的，所述抗PD-1/PD-L1抗体为帕博利珠单抗(pembrolizumab)、替雷利珠单抗(Tislelizumab)、尼伏单抗(Nivolumab)、特瑞普利单抗(Toripalimab)、阿替利珠单抗(Atezolizumab)、度伐单抗(durvalumab)、阿维单抗(Avelumab)、卡瑞利珠单抗(Camrelizumab)、信迪利单抗(Sintilimab)、西米普利单抗(Cemiplimab)、恩沃利单抗(envafolimab)、BMS-936559、JS003、SHR-1316、GS-4224、AN-4005或者MX-10181。
10.如实施方案1-8任一项所述的用途、药物组合产品或者方法，其中所述免疫检查点抑制剂为PD-1/PD-L1小分子抑制剂，进一步的，所述PD-1/PD-L1小分子抑制剂为INCB-086550、拉泽替尼(Lazertinib)、IMMH-010、CA-170、ABSK043或者RRx-001。
11.如实施方案1-8任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述免疫检查点抑制剂为TIGIT抑制剂，进一步的，所述TIGIT抑制剂为欧司珀利单抗(Ociperlimab/BGB-A1217)、维博利单抗(Vibostolimab)、domvanalimab(AB154)、替瑞利尤单抗(Tiragolumab)、Belrestotug、艾替利单抗(Etigilimab)、ONO-4686、JS-006、AZD-2936、HLX-301、SEA-TGT、M-6223、IBI-939、COM-902、AB-308、AGEN-1777、AK-127、BAT-6021、BAT-6005、 ASP-8374、PM-1022、BMS-986207、HB0036或IBI-321。
12.如实施方案1-4任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
13.如实施方案1-4任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Almonertinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
14.如实施方案1-4任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Alflutinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
15.如实施方案1-14任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，所述FAK抑制剂、所述表皮生长因子受体酪氨酸激酶抑制剂和所述免疫检查点抑制剂被同时或依次施用于所述对象。
16.如实施方案1-15任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述肿瘤为膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤，神经胶质瘤以及恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、 慢性粒细胞白血病(CML)；优选地，所述肿瘤为乳腺癌、卵巢癌、结肠癌(包括结直肠癌)、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤和胰腺癌。
17.如实施方案1-16任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述肿瘤为肺癌(包括小细胞肺癌和非小细胞肺癌)或结肠癌(包括结直肠癌)，尤其是非小细胞肺癌或结肠癌(包括结直肠癌)。
18.一种试剂盒或药学上可接受的组合物，其包括：
(a)FAK抑制剂；
(b)表皮生长因子受体酪氨酸激酶抑制剂；和
(c)免疫检查点抑制剂。
19.如实施方案18所述的试剂盒或组合物，其中所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、APG-2449、AMP945、AMP886或其药学上可接受的盐，优选为IN10018、AMP945、Defactinib、或其药学上可接受的盐，进一步优选为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐，所述IN10018结构如下：
20.如实施方案18-19所述的试剂盒或组合物，其中所述表皮生长因子受体酪氨酸激酶抑制剂为吉非替尼(Gefitinib)、厄洛替尼(Erlotinib)、埃克替尼(Icotinib)、阿法替尼(Afatinib)、达克替尼(Dacomitinib)、克唑替尼(Crizotinib)、Osimertinib(奥希替尼，AZD9291)、阿美替尼(Almonertinib)、艾氟替尼(Alflutinib，又名伏美替尼(Furmonertinib))、EAI045、JBJ-04-125-02、BLU-945、BLU-701、TQB3804、BBT-176、ES-072、BPI-361175、CH7233163或其药学上可接受的盐。
21.如实施方案18-20任一项所述的试剂盒或组合物，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib、Almonertinib、Alflutinib或其药学上可接受的盐。
22.如实施方案18-21任一项所述的试剂盒或组合物，其中所述表皮生 长因子受体酪氨酸激酶抑制剂为Osimertinib或其药学上可接受的盐。
23.如实施方案18-21任一项所述的试剂盒或组合物，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Almonertinib或其药学上可接受的盐。
24.如实施方案18-21任一项所述的组合物，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Alflutinib或其药学上可接受的盐。
25.如实施方案18-24任一项所述的试剂盒或组合物，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体、PD-1/PD-L1小分子抑制剂或者TIGIT抑制剂。
26.如实施方案18-25任一项所述的试剂盒或组合物，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体，进一步的，所述抗PD-1/PD-L1抗体为帕博利珠单抗(pembrolizumab)、替雷利珠单抗(Tislelizumab)、尼伏单抗(Nivolumab)、特瑞普利单抗(Toripalimab)、阿替利珠单抗(Atezolizumab)、度伐单抗(durvalumab)、阿维单抗(Avelumab)、卡瑞利珠单抗(Camrelizumab)、信迪利单抗(Sintilimab)、西米普利单抗(Cemiplimab)、恩沃利单抗(envafolimab)、BMS-936559、JS003、SHR-1316、GS-4224、AN-4005或者MX-10181。
27.如实施方案18-25任一项所述的用途、药物组合产品或者方法，其中所述免疫检查点抑制剂为PD-1/PD-L1小分子抑制剂，进一步的，所述PD-1/PD-L1小分子抑制剂为INCB-086550、拉泽替尼(Lazertinib)、IMMH-010、CA-170、ABSK043或者RRx-001。
28.如实施方案18-25任一项所述的试剂盒或组合物，其中所述免疫检查点抑制剂为TIGIT抑制剂，进一步的，所述TIGIT抑制剂为欧司珀利单抗(Ociperlimab/BGB-A1217)、维博利单抗(Vibostolimab)、domvanalimab(AB154)、替瑞利尤单抗(Tiragolumab)、Belrestotug、艾替利单抗(Etigilimab)、ONO-4686、JS-006、AZD-2936、HLX-301、SEA-TGT、M-6223、IBI-939、COM-902、AB-308、AGEN-1777、AK-127、BAT-6021、BAT-6005、ASP-8374、PM-1022、BMS-986207、HB0036或IBI-321。
29.如实施方案18-21任一项所述的试剂盒或组合物，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
30.如实施方案18-21任一项所述的试剂盒或组合物，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Almonertinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
31.如实施方案18-21任一项所述的试剂盒或组合物，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Alflutinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
32.如实施方案18-31任一项所述的试剂盒或组合物，其中所述组合物用于药物。
33.如实施方案32所述的试剂盒或组合物，其中所述药物用于治疗肿瘤，所述肿瘤为膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、神经胶质瘤或者恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)；优选地，所述肿瘤为乳腺癌、卵巢癌、结肠癌(包括结直肠癌)、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤和胰腺癌。
34.如实施方案33所述的试剂盒或组合物，其中所述肿瘤为肺癌(包括小细胞肺癌和非小细胞肺癌)或结肠癌(包括结直肠癌)，尤其是非小细胞肺癌或结肠癌(包括结直肠癌)。
35.一种在对象中治疗肿瘤的方法，其中所述方法包括向所述对象施用治疗有效量的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂。
36.如实施方案35所述的方法，其中所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、APG-2449、AMP945、 AMP886或其药学上可接受的盐，优选为IN10018、AMP945、Defactinib、或其药学上可接受的盐，进一步优选为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐，所述IN10018结构如下：
37.如实施方案35-36所述的方法，其中所述表皮生长因子受体酪氨酸激酶抑制剂为吉非替尼(Gefitinib)、厄洛替尼(Erlotinib)、埃克替尼(Icotinib)、阿法替尼(Afatinib)、达克替尼(Dacomitinib)、克唑替尼(Crizotinib)、Osimertinib(奥希替尼，AZD9291)、阿美替尼(Almonertinib)、艾氟替尼(Alflutinib，又名伏美替尼(Furmonertinib))、EAI045、JBJ-04-125-02、BLU-945、BLU-701、TQB3804、BBT-176、ES-072、BPI-361175、CH7233163或其药学上可接受的盐。
38.如实施方案35-37任一项所述的方法，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib、Almonertinib、Alflutinib或其药学上可接受的盐。
39.如实施方案35-38任一项所述的方法，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib或其药学上可接受的盐。
40.如实施方案35-38任一项所述的方法，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Almonertinib或其药学上可接受的盐。
41.如实施方案35-38任一项所述的方法，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Alflutinib或其药学上可接受的盐。
42.如实施方案35-41任一项所述的方法，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体、PD-1/PD-L1小分子抑制剂或者TIGIT抑制剂。
43.如实施方案35-42任一项所述的方法，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体，进一步的，所述抗PD-1/PD-L1抗体为帕博利珠单抗(pembrolizumab)、替雷利珠单抗(Tislelizumab)、尼伏单抗(Nivolumab)、特瑞普利单抗(Toripalimab)、阿替利珠单抗(Atezolizumab)、度伐单抗(durvalumab)、阿维单抗(Avelumab)、卡瑞利珠单抗(Camrelizumab)、信迪利单抗(Sintilimab)、 西米普利单抗(Cemiplimab)、恩沃利单抗(envafolimab)、BMS-936559、JS003、SHR-1316、GS-4224、AN-4005或者MX-10181。
44.如实施方案35-42任一项所述的用途、药物组合产品或者方法，其中所述免疫检查点抑制剂为PD-1/PD-L1小分子抑制剂，进一步的，所述PD-1/PD-L1小分子抑制剂为INCB-086550、拉泽替尼(Lazertinib)、IMMH-010、CA-170、ABSK043或者RRx-001。
45.如实施方案35-42任一项所述的方法，其中所述免疫检查点抑制剂为TIGIT抑制剂，进一步的，所述TIGIT抑制剂为欧司珀利单抗(Ociperlimab/BGB-A1217)、维博利单抗(Vibostolimab)、domvanalimab(AB154)、替瑞利尤单抗(Tiragolumab)、Belrestotug、艾替利单抗(Etigilimab)、ONO-4686、JS-006、AZD-2936、HLX-301、SEA-TGT、M-6223、IBI-939、COM-902、AB-308、AGEN-1777、AK-127、BAT-6021、BAT-6005、ASP-8374、PM-1022、BMS-986207、HB0036或IBI-321。
46.如实施方案35-38任一项所述的方法，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
47.如实施方案35-38任一项所述的方法，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Almonertinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
48.如实施方案35-38任一项所述的方法，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Alflutinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
49.如实施方案35-48任一项所述的方法，所述FAK抑制剂、所述表皮生长因子受体酪氨酸激酶抑制剂和所述免疫检查点抑制剂被同时或依次施用于所述对象。
50.如实施方案35-49任一项所述的方法，其中所述肿瘤为膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横 纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、神经胶质瘤或者恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)；优选地，所述肿瘤为乳腺癌、卵巢癌、结肠癌(包括结直肠癌)、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤和胰腺癌。
51.如实施方案35-50任一项所述的方法，其中所述肿瘤为肺癌(包括小细胞肺癌和非小细胞肺癌)或结肠癌(包括结直肠癌)，尤其为非小细胞肺癌或结肠癌(包括结直肠癌)。
52.FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其用于通过在对象中增加免疫原性细胞死亡来治疗肿瘤的方法。
53.如实施方案52所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、APG-2449、AMP945、AMP886或其药学上可接受的盐，优选为IN10018、AMP945、Defactinib、或其药学上可接受的盐，进一步优选为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐，所述IN10018结构如下：
54.如实施方案52-53所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述表皮生长因子受体酪氨酸激酶抑制剂为吉非替尼(Gefitinib)、厄洛替尼(Erlotinib)、埃克替尼(Icotinib)、阿法替尼(Afatinib)、达克替尼(Dacomitinib)、克唑替尼(Crizotinib)、Osimertinib(奥希替尼，AZD9291)、阿美替尼(Almonertinib)、艾氟替尼(Alflutinib，又名伏美 替尼(Furmonertinib))、EAI045、JBJ-04-125-02、BLU-945、BLU-701、TQB3804、BBT-176、ES-072、BPI-361175、CH7233163或其药学上可接受的盐。
55.如实施方案52-54任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib、Almonertinib、Alflutinib或其药学上可接受的盐。
56.如实施方案52-55任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib或其药学上可接受的盐。
57.如实施方案52-55任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Almonertinib或其药学上可接受的盐。
58.如实施方案52-55任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Alflutinib或其药学上可接受的盐。
59.如实施方案52-58任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体、PD-1/PD-L1小分子抑制剂或者TIGIT抑制剂。
60.如实施方案52-59任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体，进一步的，所述抗PD-1/PD-L1抗体为帕博利珠单抗(pembrolizumab)、替雷利珠单抗(Tislelizumab)、尼伏单抗(Nivolumab)、特瑞普利单抗(Toripalimab)、阿替利珠单抗(Atezolizumab)、度伐单抗(durvalumab)、阿维单抗(Avelumab)、卡瑞利珠单抗(Camrelizumab)、信迪利单抗(Sintilimab)、西米普利单抗(Cemiplimab)、恩沃利单抗(envafolimab)、BMS-936559、JS003、SHR-1316、GS-4224、AN-4005或者MX-10181。
61.如实施方案52-59任一项所述的用途、药物组合产品或者方法，其中所述免疫检查点抑制剂为PD-1/PD-L1小分子抑制剂，进一步的，所述PD-1/PD-L1小分子抑制剂为INCB-086550、拉泽替尼(Lazertinib)、IMMH-010、CA-170、ABSK043或者RRx-001。
62.如实施方案52-59任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述免疫检查点抑制剂为TIGIT 抑制剂，进一步的，所述TIGIT抑制剂为欧司珀利单抗(Ociperlimab/BGB-A1217)、维博利单抗(Vibostolimab)、domvanalimab(AB154)、替瑞利尤单抗(Tiragolumab)、Belrestotug、艾替利单抗(Etigilimab)、ONO-4686、JS-006、AZD-2936、HLX-301、SEA-TGT、M-6223、IBI-939、COM-902、AB-308、AGEN-1777、AK-127、BAT-6021、BAT-6005、ASP-8374、PM-1022、BMS-986207、HB0036或IBI-321。
63.如实施方案52-55任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
64.如实施方案52-55任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Almonertinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
65.如实施方案52-55任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Alflutinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
66.如实施方案52-65任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，所述FAK抑制剂、所述表皮生长因子受体酪氨酸激酶抑制剂和所述免疫检查点抑制剂被同时或依次施用于所述对象。
67.如实施方案52-66任一项所述的FAK抑制剂表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述肿瘤为膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、 子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、神经胶质瘤或者恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)；优选地，所述肿瘤为乳腺癌、卵巢癌、结肠癌(包括结直肠癌)、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤和胰腺癌。
68.如实施方案52-67任一项所述的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂，其中所述肿瘤为肺癌(包括小细胞肺癌和非小细胞肺癌)或结肠癌(包括结直肠癌)，尤其是非小细胞肺癌或结肠癌(包括结直肠癌)。
69.一种通过在对象中增加免疫原性细胞死亡来治疗肿瘤的方法，其中所述方法包括向所述对象施用治疗有效量的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂。
70.如实施方案69所述的方法，其中所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、APG-2449、AMP945、AMP886或其药学上可接受的盐，优选为IN10018、AMP945、Defactinib、或其药学上可接受的盐，进一步优选为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐，所述IN10018结构如下：
71.如实施方案69-70所述的方法，其中所述表皮生长因子受体酪氨酸激酶抑制剂为吉非替尼(Gefitinib)、厄洛替尼(Erlotinib)、埃克替尼(Icotinib)、阿法替尼(Afatinib)、达克替尼(Dacomitinib)、克唑替尼(Crizotinib)、Osimertinib(奥希替尼，AZD9291)、阿美替尼(Almonertinib)、艾氟替尼(Alflutinib，又名伏美替尼(Furmonertinib))、EAI045、JBJ-04-125-02、BLU-945、BLU-701、TQB3804、BBT-176、ES-072、BPI-361175、CH7233163或其药学上可接受的盐。
72.如实施方案69-71任一项所述的方法，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib、Almonertinib、Alflutinib或其药学上可接受的盐。
73.如实施方案69-72任一项所述的方法，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib或其药学上可接受的盐。
74.如实施方案69-72任一项所述的方法，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Almonertinib或其药学上可接受的盐。
75.如实施方案69-72任一项所述的方法，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Alflutinib或其药学上可接受的盐。
76.如实施方案69-75任一项所述的方法，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体、PD-1/PD-L1小分子抑制剂或者TIGIT抑制剂。
77.如实施方案69-76任一项所述的方法，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体，进一步的，所述抗PD-1/PD-L1抗体为帕博利珠单抗(pembrolizumab)、替雷利珠单抗(Tislelizumab)、尼伏单抗(Nivolumab)、特瑞普利单抗(Toripalimab)、阿替利珠单抗(Atezolizumab)、度伐单抗(durvalumab)、阿维单抗(Avelumab)、卡瑞利珠单抗(Camrelizumab)、信迪利单抗(Sintilimab)、西米普利单抗(Cemiplimab)、恩沃利单抗(envafolimab)、BMS-936559、JS003、SHR-1316、GS-4224、AN-4005或者MX-10181。
78.如实施方案69-76任一项所述的用途、药物组合产品或者方法，其中所述免疫检查点抑制剂为PD-1/PD-L1小分子抑制剂，进一步的，所述PD-1/PD-L1小分子抑制剂为INCB-086550、拉泽替尼(Lazertinib)、IMMH-010、CA-170、ABSK043或者RRx-001。
79.如实施方案69-76任一项所述的方法，其中所述免疫检查点抑制剂为TIGIT抑制剂，进一步的，所述TIGIT抑制剂为欧司珀利单抗(Ociperlimab/BGB-A1217)、维博利单抗(Vibostolimab)、domvanalimab(AB154)、替瑞利尤单抗(Tiragolumab)、Belrestotug、艾替利单抗(Etigilimab)、ONO-4686、JS-006、AZD-2936、HLX-301、SEA-TGT、M-6223、IBI-939、COM-902、AB-308、AGEN-1777、AK-127、BAT-6021、BAT-6005、ASP-8374、PM-1022、BMS-986207、HB0036或IBI-321。
80.如实施方案69-72任一项所述的方法，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂 为Osimertinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
81.如实施方案69-72任一项所述的方法，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Almonertinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
82.如实施方案69-72任一项所述的方法，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Alflutinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
83.如实施方案69-82任一项所述的方法，所述FAK抑制剂、所述表皮生长因子受体酪氨酸激酶抑制剂和所述免疫检查点抑制剂被同时或依次施用于所述对象。
84.如实施方案69-83任一项所述的方法，其中所述肿瘤为膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、神经胶质瘤或者恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)；优选地，所述肿瘤为乳腺癌、卵巢癌、结肠癌(包括结直肠癌)、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤和胰腺癌。
85.如实施方案69-84任一项所述的方法，其中所述肿瘤为肺癌(包括小细胞肺癌和非小细胞肺癌)或结肠癌(包括结直肠癌)，尤其是非小细胞肺癌或结肠癌(包括结直肠癌)。
86.FAK抑制剂在制备用于在对象中治疗肿瘤的药物中的用途，其中将所述FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制 剂施用于所述对象。
87.表皮生长因子受体酪氨酸激酶抑制剂在制备用于在对象中治疗肿瘤的药物中的用途，其中将FAK抑制剂，所述表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂施用于所述对象。
88.免疫检查点抑制剂在制备用于在对象中治疗肿瘤的药物中的用途，其中将FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和所述免疫检查点抑制剂施用于所述对象。
89.FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂在制备用于治疗肿瘤的联用药物中的用途。
90.FAK抑制剂在制备用于与表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂治疗肿瘤的联用药物中的用途。
91.表皮生长因子受体酪氨酸激酶抑制剂在制备用于与FAK抑制剂和免疫检查点抑制剂治疗肿瘤的联用药物中的用途。
92.免疫检查点抑制剂在制备用于与FAK抑制剂和表皮生长因子受体酪氨酸激酶抑制剂治疗肿瘤的联用药物中的用途。
93.FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂在制备用于联合治疗肿瘤的药物中的用途。
94.FAK抑制剂在制备用于与表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂联合治疗肿瘤的药物中的用途。
95.表皮生长因子受体酪氨酸激酶抑制剂在制备用于与FAK抑制剂和免疫检查点抑制剂联合治疗肿瘤的药物中的用途。
96.免疫检查点抑制剂在制备用于与FAK抑制剂和表皮生长因子受体酪氨酸激酶抑制剂联合治疗肿瘤的药物中的用途。
97.如实施方案86-96任一项所述的用途，其中所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、APG-2449、AMP945、AMP886或其药学上可接受的盐，优选为IN10018、AMP945、Defactinib、或其药学上可接受的盐，进一步优选为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐，所述IN10018结构如下：
98.如实施方案86-97任一项所述的用途，其中所述表皮生长因子受体酪氨酸激酶抑制剂为吉非替尼(Gefitinib)、厄洛替尼(Erlotinib)、埃克替尼(Icotinib)、阿法替尼(Afatinib)、达克替尼(Dacomitinib)、克唑替尼(Crizotinib)、Osimertinib(奥希替尼，AZD9291)、阿美替尼(Almonertinib)、艾氟替尼(Alflutinib，又名伏美替尼(Furmonertinib))、EAI045、JBJ-04-125-02、BLU-945、BLU-701、TQB3804、BBT-176、ES-072、BPI-361175、CH7233163或其药学上可接受的盐。
99.如实施方案86-98任一项所述的用途，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib、Almonertinib、Alflutinib或其药学上可接受的盐。
100.如实施方案86-99任一项所述的用途，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib或其药学上可接受的盐。
101.如实施方案86-99任一项所述的用途，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Almonertinib或其药学上可接受的盐。
102.如实施方案86-99任一项所述的用途，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Alflutinib或其药学上可接受的盐。
103.如实施方案86-102任一项所述的用途，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体、PD-1/PD-L1小分子抑制剂或者TIGIT抑制剂。
104.如实施方案86-103任一项所述的用途，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体，进一步的，所述抗PD-1/PD-L1抗体为帕博利珠单抗(pembrolizumab)、替雷利珠单抗(Tislelizumab)、尼伏单抗(Nivolumab)、特瑞普利单抗(Toripalimab)、阿替利珠单抗(Atezolizumab)、度伐单抗(durvalumab)、阿维单抗(Avelumab)、卡瑞利珠单抗(Camrelizumab)、信迪利单抗(Sintilimab)、西米普利单抗(Cemiplimab)、恩沃利单抗(envafolimab)、BMS-936559、JS003、SHR-1316、GS-4224、AN-4005或者MX-10181。
105.如实施方案86-103任一项所述的用途、药物组合产品或者方法， 其中所述免疫检查点抑制剂为PD-1/PD-L1小分子抑制剂，进一步的，所述PD-1/PD-L1小分子抑制剂为INCB-086550、拉泽替尼(Lazertinib)、IMMH-010、CA-170、ABSK043或者RRx-001。
106.如实施方案86-103任一项所述的用途，其中所述免疫检查点抑制剂为TIGIT抑制剂，进一步的，所述TIGIT抑制剂为欧司珀利单抗(Ociperlimab/BGB-A1217)、维博利单抗(Vibostolimab)、domvanalimab(AB154)、替瑞利尤单抗(Tiragolumab)、Belrestotug、艾替利单抗(Etigilimab)、ONO-4686、JS-006、AZD-2936、HLX-301、SEA-TGT、M-6223、IBI-939、COM-902、AB-308、AGEN-1777、AK-127、BAT-6021、BAT-6005、ASP-8374、PM-1022、BMS-986207、HB0036或IBI-321。
107.如实施方案86-99任一项所述的用途，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
108.如实施方案86-99任一项所述的用途，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Almonertinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
109.如实施方案86-99任一项所述的用途，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Alflutinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
110.如实施方案86-109任一项所述的用途，所述FAK抑制剂、所述表皮生长因子受体酪氨酸激酶抑制剂和所述免疫检查点抑制剂被同时或依次施用于所述对象。
111.如实施方案86-110任一项所述的用途，其中所述肿瘤为膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、 神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、神经胶质瘤或者恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)；优选地，所述肿瘤为乳腺癌、卵巢癌、结肠癌(包括结直肠癌)、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤和胰腺癌。
112.如实施方案86-111任一项所述的用途，其中所述肿瘤为肺癌(包括小细胞肺癌和非小细胞肺癌)或结肠癌(包括结直肠癌)，尤其是非小细胞肺癌或结肠癌(包括结直肠癌)。
实施例
提供下面的实施例以进一步阐述本发明。应理解，这些实施例仅用于举例说明本发明，而不用于限制本发明的范围。
下列实施例中未注明具体条件的实验方法均可以按照这类反应的常规条件进行或者按照制造厂商所建议的条件进行。以下实施例中所用的实验材料和试剂如无特别说明均可从市售渠道获得。
实施例中所用的缩写含义如下：

实施例1：IN10018和AZD9291在肺癌KPL细胞中的研究
KPL细胞(中科院细胞所)用RPMI 1640(上海元培，分类号:L210KJ,批号:F210916)+10％FBS(Gibco，分类号:10099-141c,批号:2158737cp)培养，传代两次，当细胞状况良好时，将培养液放置在24孔板中。细胞铺展24小时后，设四组，第一组为阴性对照组，加入培养基，第二组为IN10018，浓度为10μM，第三组为AZD9291，浓度为10μM，第四组为IN10018和AZD9291联用组，分别为IN10018(10μM)和AZD9291(10μM)。将药物混合，并在37℃的5％CO2培养箱中培养48小时。
药物作用48小时后，对细胞进行显微镜观察和拍照，并保存照片。然后收集细胞进行流动分析，并用流动缓冲液(PBS+2％FBS)洗涤细胞两次，将0.5μl共轭GRP94多克隆抗体(Proteintech，分类号：CL488-14700，批号：21006579)添加到每个孔中，混合，在4℃且避光的 条件下孵育20min。20min后，用流动缓冲液(PBS+2％FBS)清洗细胞两次。然后在流式细胞仪上进行分析。
显微镜下观察细胞，AZD9291单药组细胞状态较差，双药联合组细胞状态最差，细胞死亡较多，而阴性对照组和IN10018组细胞状态较好。流式细胞仪结果显示，双药联合组的CRT阳性率和Annexin V阳性率高于单药组。详见图1、图2a和图2b。
实施例2：IN10018和Almonertinib在肺癌KPL细胞中的研究
KPL细胞(中科院细胞所)用RPMI 1640(上海元培，分类号:L210KJ,批号:F210916)+10％FBS(Gibco，分类号:10099-141c,批号:2158737cp)培养，传代两次，当细胞状况良好时，将培养液放置在24孔板中。细胞铺展24小时后，设四组，第一组为阴性对照组，加入培养基，第二组为IN10018，浓度为10μM，第三组为Almonertinib，浓度为4.7μM，第四组为IN10018和Almonertinib联用组，分别为IN10018(10μM)和Almonertinib(4.7μM)。将药物混合，并在37℃的5％CO2培养箱中培养48小时。
药物作用48小时后，对细胞进行显微镜观察和拍照，并保存照片。然后收集细胞进行流动分析，并用流动缓冲液(PBS+2％FBS)洗涤细胞两次，将0.5μl共轭GRP94多克隆抗体(Proteintech，分类号：CL488-14700，批号：21006579)添加到每个孔中，混合，在4℃且避光的条件下孵育20min。20min后，用流动缓冲液(PBS+2％FBS)清洗细胞两次。然后在流式细胞仪上进行分析。
显微镜下观察细胞，Almonertinib单药组细胞状态较差，双药联合组细胞状态最差，细胞死亡较多，而阴性对照组和IN10018组细胞状态较好。流式细胞仪结果显示，双药联合组的CRT阳性率和Annexin V阳性率高于单药组。详见图3、图4a和图4b。
实施例3：IN10018和Alflutinib在肺癌KPL细胞中的研究
KPL细胞(中科院细胞所)用RPMI 1640(上海元培，分类号:L210KJ,批号:F210916)+10％FBS(Gibco，分类号:10099-141c,批号:2158737cp)培养，传代两次，当细胞状况良好时，将培养液放置在24孔板中。细胞铺展24小时后，设四组，第一组为阴性对照组，加入培养基，第二组为IN10018， 浓度为10μM，第三组为Alflutinib，浓度为4.7μM，第四组为IN10018和Alflutinib联用组，分别为IN10018(10μM)和Alflutinib(4.7μM)。将药物混合，并在37℃的5％CO2培养箱中培养48小时。
药物作用48小时后，对细胞进行显微镜观察和拍照，并保存照片。然后收集细胞进行流动分析，并用流动缓冲液(PBS+2％FBS)洗涤细胞两次，将0.5μl共轭GRP94多克隆抗体(Proteintech，分类号：CL488-14700，批号：21006579)添加到每个孔中，混合，在4℃且避光的条件下孵育20min。20min后，用流动缓冲液(PBS+2％FBS)清洗细胞两次。然后在流式细胞仪上进行分析。
显微镜下观察细胞，Alflutinib单药组细胞状态较差，双药联合组细胞状态最差，细胞死亡较多，而阴性对照组和IN10018组细胞状态较好。流式细胞仪结果显示，双药联合组的CRT阳性率和Annexin V阳性率高于单药组。详见图5、图6a和图6b。
实施例4：FAK靶点抑制增强非小细胞肺癌细胞针对AZD9291免疫原性细胞死亡的研究
实验方案：
1.FAK沉默，联合AZD9291促进肺癌细胞HCC827凋亡
体外实验利用siRNA技术降低FAK的表达水平，与AZD9291联合使用48小时；利用流式细胞仪，检测对照和FAK沉默的细胞凋亡
2.FAK沉默，联合AZD9291能够有效促进肺癌细胞HCC827产生ICD作用
体外实验利用siRNA技术降低FAK的表达水平，与AZD9291联合使用48小时；利用流式细胞仪，检测ICD的主要靶标Calreticulin的表达情况。
实验抗体：重组Alexa荧光Anti-Calreticulin(抗-钙网蛋白)抗体(Abcam，ab196159)，Annexin V细胞凋亡检测试剂盒(Invitrogen,A35110)。
FAK siRNA由吉玛基因提供，序列如下表所示：
F：CCUGUAUGCCUAUCAGCUUTT；
R：AAGCUGAUAGGCAUACAGGTT。
实验仪器：
荧光显微镜(Olympus U-HGLGPS)。化学发光成像仪(BIORAD chemidoc touch)
实验结果：
1、FAK沉默后联合AZD9291显著增加HCC827细胞凋亡
分别使用终浓度为50nM的对照siRNA或FAK siRNA转染HCC827细胞。转染24小时后，给予0.3nM AZD9291处理48小时，使用Annexin V试剂盒进行细胞染色并使用流式细胞仪进行检测。统计细胞早期和晚期凋亡数值与DMSO对照组进行比较并使用Graphpad 8.0进行绘图。
结果显示FAK沉默组与对照组比较，经过AZD9291处理后，晚期凋亡被显著的增强了，如图7所示。
2、FAK沉默后联合AZD9291显著上调了ICD靶点Calreticulin释放及暴露
分别使用终浓度为50nM的对照siRNA或FAK siRNA转染HCC827细胞。转染24小时后，给予0.3nM AZD9291处理48小时，用Calreticulin抗体进行荧光染色，流式细胞仪对染色结果经行分析。FlowJo软件结果统计显示：FAK沉默后联合AZD9291，Calreticulin的释放及暴露均被显著的增强了，如图8所示。
实施例5：IN10018增强非小细胞肺癌细胞针对AZD9291免疫原性细胞死亡的研究
实验方案：
1.IN10018联合AZD9291产生协同肿瘤杀伤作用，并诱导细胞凋亡
体外实验检测AZD9291在肺癌细胞HCC827中的杀伤曲线并确定IC50值。探索不同剂量AZD9291分别与3μM IN10018和5μM IN10018合用检测细胞毒作用。同时利用流式细胞仪，采用Annexin V染色检测药物单用与合用细胞凋亡反应。
2.IN10018联合AZD9291能有效促进肺癌细胞HCC827产生ICD作用
体外实验IN10018与AZD9291联用6小时检测内质网应激信号通路激活；两药联用48小时检测ICD的主要靶标Calreticulin的表达情况。
实验抗体：
重组Alexa647荧光Anti-Calreticulin(抗-钙网蛋白)抗体(Abcam， ab196159)，FAK抗体(CST，3285S)，Phospho-eIF2α(Ser51)(CST，3398)，eif2α(CST，5324)，DDIT3(HUABIO，ET1703-05)，HRP标记Alpha tubulin(α-微管蛋白)抗体(Proteintech，HRP-66031)，Annexin V细胞凋亡检测试剂盒(Invitrogen,A35110)
实验仪器：
荧光显微镜(Olympus U-HGLGPS)。化学发光成像仪(BIORAD chemidoc touch)。
实验结果：
1、IN10018联合AZD9291产生协同肿瘤杀伤作用
以5000个HCC827细胞/孔铺设96孔细胞板，24小时后加入定浓度的IN10018联合不同浓度的AZD9291。IN10018的浓度为3μM和5μM；AZD9291最高浓度为1μM以3倍浓度梯度稀释共设9个浓度。药物与细胞共同孵育72小时后每孔加入10μM CCK8溶液，于37℃、5％二氧化碳孵箱孵育2小时后，使用酶标仪设定吸光度波长为450nm进行读值。读取的数值与DMSO对照组进行比较并使用Graphpad 8.0进行绘图。结果显示不同剂量下AZD9291分别与3μM IN10018和5μM IN10018合用48小时对非小细胞肺癌细胞HCC827的杀伤协同作用，如图9所示。
2、IN10018联合AZD9291促进细胞凋亡
使用0.3nM AZD9291与3μM IN10018联合处理HCC827细胞，48小时后收取细胞使用Annexin V试剂盒进行细胞染色并使用流式细胞仪进行检测。统计细胞早期和晚期凋亡数值与DMSO对照组进行比较并使用Graphpad 8.0进行绘图。结果显示合用组与单药组比较，早期及晚期凋亡均被显著的增强了，如图10所示。
3、IN10018联合AZD9291能够显著上调了HCC827的内质网应激
分别用0.1nM和0.3nM AZD9291与3μM IN10018合用处理HCC827细胞，6小时后检测内质网应激相关蛋白Phospho-eIF2α(Ser51)、DDIT3的蛋白水平表达。结果显示合用组与单药组比较，Phospho-eIF2α(Ser51)、DDIT3的表达水平显著上调，说明两药合用内质网压力显著上调，如图11所示。
4、AZD9291与IN10018合用增强了ICD靶点Calreticulin释放及暴露
0.3nM AZD9291及3μM IN10018合用处理HCC827细胞，48小时后使用Calreticulin抗体进行荧光染色，流式细胞仪对染色结果经行分析。FlowJo 软件结果统计显示合用组与单药组比较，Calreticulin的释放及暴露均被显著的增强了，如图12所示。
实施例6：AZD9291在结肠癌CT26细胞BALB/c小鼠皮下同种移植瘤模型中的体内抗肿瘤药效研究
试验材料：
小鼠：6-8周龄的雌性BALB/c小鼠购自上海灵畅生物科技有限责任公司。动物到达后在实验环境适应性饲养后开始实验。动物在SPF级动物房以IVC(独立送风系统)笼具饲养(每笼5只)。所有笼具、垫料及饮水在使用前均需灭菌。所有实验人员在动物房操作时应穿着防护服和乳胶手套。每笼动物信息卡应注明笼内动物数目，性别，品系，接收日期，给药方案，实验编号，组别以及实验开始日期。笼具、饲料及饮水每周更换两次。饲养环境及光照情况如下：
温度：20-26℃
湿度：40-70％
光照周期：12小时光照，12小时无光照
笼具：以聚碳酸酯制成，体积300mm×180mm×150mm。垫料为玉米芯，每周更换两次。
食物：实验动物在整个实验阶段中可自由进食(辐照灭菌，干颗粒状食物)。
饮水：实验动物可自由饮用灭菌水。
动物标识：实验动物以耳标进行标识。
化合物信息见表1
表1.化合物信息

结直肠癌细胞CT26(来源南京科佰生物科技有限公司，货号：CBP60043)由应世生物科技(南京)有限公司维持传代。细胞体外单层培养，培养条件为RPMI-1640培养基中加10％胎牛血清，37℃5％CO2培养箱中培养。一周两到三次用胰酶-EDTA进行常规消化处理传代。当细胞处于指数生长期，饱和度为80％-90％时，收取细胞，计数后接种。
细胞接种及分组
将0.1mL含有3×105个细胞的细胞悬液皮下接种于每只小鼠的右后背。当肿瘤体积达到～63mm3时(细胞接种后第12天)，根据肿瘤体积进行随机分组给药，分组信息见表2。
表2.受试物对CT26小鼠移植瘤模型的给药方案
注：1.每组小鼠数目；
2.给药体积：根据小鼠体重10mL/kg，如果体重下降超过15％，动物停止给药；待体重恢复至降低10％，再恢复给药。
受试物的配制
详见表3
表3.受试物配置方法
实验动物日常观察
本实验方案的拟定及任何修改均通过了云桥生物IACUC的评估核准。实验动物的使用及福利遵照AAALAC的规定执行。每天监测动物的健康状况及死亡情况，例行检查包括观察肿瘤生长和药物治疗对动物日常行为表现的影响如行为活动，摄食摄水量(仅目测)，体重变化，外观体征或其它不正常情况。基于各组动物数量记录了组内动物死亡数和副作用。
实验终止
若动物健康状况持续恶化，或瘤体积超过3,000mm3，或有严重疾病，或疼痛，须处以安乐死。有以下情况者，通知兽医并处以安乐死：明显消瘦，体重降低大于20％；不能自由取食和饮水；对照组瘤体积平均值达到2,000mm3，实验终止。动物出现以下临床表现且持续恶化：立毛，弓背，耳、鼻、眼或足色发白，呼吸仓促，抽搐，连续腹泻，脱水，行动迟缓，发声。
肿瘤测量和实验指标
用游标卡尺测量肿瘤直径，每周测量3次。肿瘤体积的计算公式为：V＝0.5×a×b2，a和b分别表示肿瘤的长径和短径。
参照分组后第一天的肿瘤体积，根据以下公式计算肿瘤生长抑制率TGI(％)。
TGI(％)＝[1-(某给药组的平均瘤体积-该给药组开始治疗时平均瘤体积)/(溶剂对照组的平均瘤体积-溶剂对照组开始治疗时平均瘤体 积)]×100％。
统计分析
统计分析基于试验结束时肿瘤体积和肿瘤重量运用Prism Graphpad软件进行分析。多组间比较，采用Two-way ANOVA，Fisher’s LSD test法检验进行分析。P<0.05认为有显著性差异。
实验结果
受试物AZD9291和/或PD-L1抗体与IN10018联用在CT26小鼠结直肠癌细胞BALB/c小鼠皮下同种移植瘤模型中的体内药效。细胞接种后，每天观察肿瘤生长情况，接种后第12天根据肿瘤体积进行分组，入组平均肿瘤体积约为63mm3。由于肿瘤负荷，对照组在接种后的第27天，即分组给药之后的第15天安乐死，整个实验结束。
分组给药后第15天，对照组的肿瘤体积为2777.3±705.2mm3。AZD9291+IN10018(20+25mg/kg)、AZD9291+PDL1抗体(20+10mg/kg)和AZD9291+IN10018+PDL1抗体(20+25+10mg/kg)各治疗组的肿瘤体积分别为1693.4±1207.5mm3、1387.6±859.6mm3和927.2±627.9mm3，详见表4。综合肿瘤体积与对照组进行比较，AZD9291+IN10018(20+25mg/kg)、AZD9291+PDL1抗体(20+10mg/kg)和AZD9291+IN10018+PDL1抗体(20+25+10mg/kg)组的抑瘤率TGI分别为40.0％(p<0.0001)、51.2％(p<0.0001)和62.7％(p<0.0001)。详见表4。综合肿瘤体积与AZD9291+IN10018+PDL1抗体(20+25+10mg/kg)三药联用组相比较，进行统计学分析，对照组、AZD9291+IN10018(20+25mg/kg)和AZD9291+PDL1抗体(20+10mg/kg)各治疗组的P值分别为p<0.0001、p＝0.0032和p＝0.0745。各剂量组在不同时间段的肿瘤体积如图13所示。
表4：受试物对小鼠结肠癌CT26细胞的BALB/c小鼠移植瘤模型的抑瘤效果评价(基于分组给药后第15天数据)
注：1.按照分组给药后的天数来计算，数据为平均值±标准误差。
2.****：p<0.0001，vs.对照组，Two-way ANOVA。
3.**：p<0.01，****：p<0.0001，vs.AZD9291+IN10018+PDL1抗体(20+25+10mg/kg)组，Two-way ANOVA。
实验按照给药方案进行，实验过程中，每天观察动物摄食饮水等活动，每周记录3次动物体重，动物体重曲线见图14。在整个给药周期中，各组动物体重均无明显下降且状态良好。
结论
与空白对照组相比，AZD9291+IN10018(20+25mg/kg)、AZD9291+PDL1抗体(20+10mg/kg)和AZD9291+IN10018+PDL1抗体(20+25+10mg/kg)治疗组均有明显肿瘤生长抑制作用，和对照组相比都有统计学差异。综合整个给药周期，AZD9291+IN10018+PDL1抗体(20+25+10mg/kg)三药联用组相对于AZD9291+IN10018(20+25mg/kg)、AZD9291+PDL1抗体(20+10mg/kg)两药联用组的肿瘤体积一直都更小，并且和AZD9291+IN10018(20+25mg/kg)组相比均具有统计学差异，其相对于AZD9291+IN10018(20+25mg/kg)、AZD9291+PDL1抗体(20+10mg/kg)两药联用组，具有更好的抑制肿瘤生长的效果。同时，动物体重变化良好，整个给药周期的活动、饮水进食以及精神状况也未发现异常，说明动物对AZD9291+IN10018+PDL1抗体(20+25+10mg/kg)三药联用耐受。
实施例7：AZD9291在结肠癌MC38细胞C57BL/6小鼠皮下同种移植瘤模型中的体内抗肿瘤药效研究
试验材料：
小鼠：6-8周龄的雌性C57BL/6小鼠购自上海斯莱克实验动物有限责任公司。动物到达后在实验环境适应性饲养后开始实验。动物在SPF级动物房以IVC(独立送风系统)笼具饲养(每笼5只)。所有笼具、垫料及饮水在使用前均需灭菌。所有实验人员在动物房操作时应穿着防护服和乳胶手套。每笼动物信息卡应注明笼内动物数目，性别，品系，接收日期，给药方案，实验编号，组别以及实验开始日期。笼具、饲料及饮水每周更换两次。饲养环境及光照情况如下：
温度：20-26℃
湿度：40-70％
光照周期：12小时光照，12小时无光照
笼具：以聚碳酸酯制成，体积300mm×180mm×150mm。垫料为玉米芯，每周更换两次。
食物：实验动物在整个实验阶段中可自由进食(辐照灭菌，干颗粒状食物)。
饮水：实验动物可自由饮用灭菌水。
动物标识：实验动物以耳标进行标识。
化合物信息见表5。
表5：化合物信息
结直肠癌细胞MC38(来源南京科佰生物科技有限公司，货号：CBP60825)由应世生物科技(南京)有限公司维持传代。细胞体外单层培养，培养条件为DMEM培养基中加10％胎牛血清，37℃5％CO2培养箱中培养。一周两到三次用胰酶-EDTA进行常规消化处理传代。当细胞处于指数生长期，饱和度为80％-90％时，收取细胞，计数后接种。
细胞接种及分组
将0.1mL含有2×105个细胞的细胞悬液皮下接种于每只小鼠的右后背。当肿瘤体积达到～70mm3时(细胞接种后第15天)，根据肿瘤体积进行随机 分组给药，分组信息见表6。
表6：受试物对MC38小鼠移植瘤模型的给药方案
注：1.每组小鼠数目；
2.给药体积：根据小鼠体重10mL/kg，如果体重下降超过15％，动物停止给药；待体重恢复至降低10％，再恢复给药。
受试物的配制
详见表7
表7：受试物配置方法
实验动物日常观察
本实验方案的拟定及任何修改均通过了云桥生物IACUC的评估核准。实验动物的使用及福利遵照AAALAC的规定执行。每天监测动物的健康状况及死亡情况，例行检查包括观察肿瘤生长和药物治疗对动物日常行为表现的影响如行为活动，摄食摄水量(仅目测)，体重变化，外观体征或其它不正常情况。基于各组动物数量记录了组内动物死亡数和副作用。
实验终止
若动物健康状况持续恶化，或瘤体积超过3,000mm3，或有严重疾病，或疼痛，须处以安乐死。有以下情况者，通知兽医并处以安乐死：明显消瘦，体重降低大于20％；不能自由取食和饮水；对照组瘤体积平均值达到2,000mm3，实验终止。动物出现以下临床表现且持续恶化：立毛，弓背，耳、鼻、眼或足色发白，呼吸仓促，抽搐，连续腹泻，脱水，行动迟缓，发声。
肿瘤测量和实验指标
用游标卡尺测量肿瘤直径，每周测量3次。肿瘤体积的计算公式为：V＝0.5×a×b2，a和b分别表示肿瘤的长径和短径。
参照分组后第一天的肿瘤体积，根据以下公式计算肿瘤生长抑制率TGI(％)。TGI(％)＝[1-(某给药组的平均瘤体积-该给药组开始治疗时平均瘤体积)/(溶剂对照组的平均瘤体积-溶剂对照组开始治疗时平均瘤体积)]×100％。
统计分析
统计分析基于试验结束时肿瘤体积和肿瘤重量运用Prism Graphpad软件进行分析。多组间比较，采用Two-way ANOVA，Fisher’s LSD test法检验进行分析。P<0.05认为有显著性差异。
实验结果
受试物AZD9291和IN10018与PD-L1抗体联用在MC38小鼠结直肠癌细胞C57BL/6小鼠皮下同种移植瘤模型中的体内药效。细胞接种后，每天观察肿瘤生长情况，接种后第15天根据肿瘤体积进行分组，入组平均肿瘤体积约为70mm3。由于肿瘤负荷，对照组在接种后的第29天，即分组给药之后的第14天安乐死，整个实验结束。
分组给药后第14天，对照组的肿瘤体积为1984.4±537.8mm3。PD-L1 抗体(5mg/kg)、AZD9291(20mg/kg)、IN10018+PD-L1抗体(25+5mg/kg)和AZD9291+IN10018+PD-L1抗体(20+25+5mg/kg)各治疗组的肿瘤体积分别为1974.8±1629.9mm3，1518.1±728.8mm3，1425.2±889.6mm3和1292.9±934.6mm3，详见表8。综合肿瘤体积与对照组进行比较，PD-L1抗体(5mg/kg)、AZD9291(20mg/kg)、IN10018+PD-L1抗体(25+5mg/kg)和AZD9291+IN10018+PD-L1抗体(20+25+5mg/kg)组的抑瘤率TGI分别为0.5％(p＝0.9728)、24.4％(p＝0.1012)、29.2％(p＝0.0498)和36.1％(p＝0.0205)。详见表8。综合肿瘤体积与AZD9291+IN10018+PD-L1抗体(20+25+5mg/kg)联用组相比较，进行统计学分析，对照组、PD-L1抗体(5mg/kg)、AZD9291(20mg/kg)和IN10018+PD-L1抗体(25+5mg/kg)各组的P值分别为p＝0.0205、p＝0.0170、p＝0.4274和p＝0.6408。各剂量组在不同时间段的肿瘤体积如图15所示。
表8：受试物对小鼠结肠癌MC38细胞的C57BL/6小鼠移植瘤模型的抑瘤效果评价(基于分组给药后第14天数据)
注：1.按照分组给药后的天数来计算，数据为平均值±标准误差。
2.*：p<0.05，vs.对照组，Two-way ANOVA。
3.*：p<0.05，vs.AZD9291+IN10018+PD-L1抗体(20+25+5mg/kg)组，Two-way ANOVA。
实验按照给药方案进行，实验过程中，每天观察动物摄食饮水等活动，每周记录3次动物体重，动物体重曲线见图16。对照组有1只小鼠，由于肿瘤破溃，在分组给药后的第11天发现死亡；AZD9291+IN10018+PD-L1抗体(20+25+5mg/kg)组有1只小鼠，由于灌胃给药的失误导致在分组给药后 的第8天死亡，其余各组的动物在整个给药周期中，体重均无明显下降且状态良好，显示出对不同给药方式的耐受。
结论
与空白对照组相比，IN10018+PD-L1抗体(25+5mg/kg)和AZD9291+IN10018+PD-L1抗体(20+25+5mg/kg)组均有明显肿瘤生长抑制作用，和对照组相比都有统计学差异。综合整个给药周期，AZD9291+IN10018+PD-L1抗体(20+25+5mg/kg)组相对于其余各治疗组的肿瘤体积一直都更小，并且和PD-L1抗体(5mg/kg)单药组相比具有统计学差异，虽然三药联用组和AZD9291(20mg/kg)和IN10018+PD-L1抗体(25+5mg/kg)组相比没有统计学差异，但是其平均瘤体积相对其余各治疗组而言一直较小，也显示AZD9291+IN10018+PD-L1抗体(20+25+5mg/kg)三药联用组具有更好的抑制肿瘤生长的效果。同时，除去因肿瘤破溃而自然死亡以及人为操作失误死亡的动物之外，其余各组动物体重变化良好，整个给药周期的活动、饮水进食以及精神状况也未发现异常，说明动物对AZD9291+IN10018+PD-L1抗体(20+25+5mg/kg)三药联用耐受。
实施例8：AZD9291和AMP945对小鼠乳腺癌4T1细胞体外免疫原性细胞死亡靶点诱导的研究
实验材料：
1)本实验所用药物
AZD9291由上海超岚化工科技中心提供,Lot No.:202012-02
AMP945由MCE提供,Lot No.:143253
2)本实验所用抗体
重组Alexa647荧光Anti-Calreticulin抗体(Abcam，Cat No.:ab196159,Lot No.:CR33676773)。Annexin V-调亡检测试剂盒(Beyotime,Cat No.:C1062L,Lot No.:122221220706)。
实验方法：
4T1细胞(来源南京科佰生物科技有限公司，货号：CBP60352)用RPMI 1640(上海元培,Cat No.:L210KJ,Lot No.:F210916)+10％FBS(Gibco,Cat No.:10099-141c,Lot No.:2158737cp)培养，培养条件为37℃、5％CO2。一周两 到三次用胰蛋白酶进行常规消化传代处理。当细胞处于指数生长期，贴壁汇合至80％-90％时，收取细胞并进行铺板。4T1细胞用胰蛋白酶进行消化后收取细胞并进行计数，根据计数结果，用RPMI1640+10％FBS对细胞进行稀释，稀释浓度为5万个细胞每毫升，然后进行12孔细胞培养板铺板，每孔铺2ml细胞悬液即10万个细胞。铺板完成后，将细胞放于37℃、5％CO2培养箱中培养。细胞铺展24小时后，设立六组，第一组为对照组，添加培养基，第二组为AMP945，浓度为3μM，第三组为AMP945，浓度为6μM，第四组为AZD9291，浓度为10μM，第五组为AMP945(3μM)与AZD9291(10μM)联用，第六组为AMP945(6μM)与AZD9291(10μM)联用。将药物混合，并在37℃下在5％CO2培养箱中培养48小时。
实验结果
药物作用48h后，收集细胞进行流动分析，并使用流动缓冲液(PBS+2％FBS)和0.5μl AF647清洗细胞两次，每个孔中加入抗钙网蛋白抗体(abcam)，混合。在4℃且避光的条件下孵育，孵育20min后加入流动缓冲液，使用膜联蛋白染色试剂盒(Beyotime)，加入195μl Annexin-V-FITC结合物，与细胞进行吹打混合后，加入5μl Annexin-V-FITC抗体，轻轻混合，最后加入10μl PI染料混合，室温避光孵育15min，将样品送至流式细胞仪上进行信号测定。流式细胞仪分析结果显示两药合用组的CRT阳性率和Annexin-V阳性率显著优于单药组及对照组，见图17a和图17b。
实施例9：Alflutinib和AMP945对小鼠乳腺癌4T1细胞体外免疫原性细胞死亡靶点诱导的研究
实验材料：
1)本实验所用药物
Alflutinib由上海MCE提供,Lot No.:33805
AMP945由MCE提供,Lot No.:143253
2)本实验所用抗体
重组Alexa647荧光Anti-Calreticulin抗体(Abcam，Cat No.:ab196159,Lot No.:CR33676773)。Annexin V-调亡检测试剂盒(Beyotime,Cat No.:C1062L,Lot No.:122221220706)。
实验方法：
4T1细胞(来源南京科佰生物科技有限公司，货号：CBP60352)用RPMI 1640(上海元培,Cat No.:L210KJ,Lot No.:F210916)+10％FBS(Gibco,Cat No.:10099-141c,Lot No.:2158737cp)培养，培养条件为37℃、5％CO2。一周两到三次用胰蛋白酶进行常规消化传代处理。当细胞处于指数生长期，贴壁汇合至80％-90％时，收取细胞并进行铺板。4T1细胞用胰蛋白酶进行消化后收取细胞并进行计数，根据计数结果，用RPMI1640+10％FBS对细胞进行稀释，稀释浓度为5万个细胞每毫升，然后进行12孔细胞培养板铺板，每孔铺2ml细胞悬液即10万个细胞。铺板完成后，将细胞放于37℃、5％CO2培养箱中培养。细胞铺展24小时后，设立六组，第一组为对照组，添加培养基，第二组为AMP945，浓度为3μM，第三组为AMP945，浓度为6μM，第四组为Alflutinib，浓度为10μM，第五组为AMP945(3μM)与Alflutinib(10μM)联用，第六组为AMP945(6μM)与Alflutinib(10μM)联用。将药物混合，并在37℃下在5％CO2培养箱中培养48小时。
实验结果
药物作用48h后，收集细胞进行流动分析，并使用流动缓冲液(PBS+2％FBS)和0.5μl AF647清洗细胞两次，每个孔中加入抗钙网蛋白抗体(abcam)，混合。在4℃且避光的条件下孵育，孵育20min后加入流动缓冲液，使用膜联蛋白染色试剂盒(Beyotime)，加入195μl Annexin-V-FITC结合物，与细胞进行吹打混合后，加入5μl Annexin-V-FITC抗体，轻轻混合，最后加入10μl PI染料混合，室温避光孵育15min，将样品送至流式细胞仪上进行信号测定。流式细胞仪分析结果显示两药合用组的CRT阳性率和Annexin-V阳性率显著优于单药组及对照组，见图18a和图18b。
实施例10：Almonertinib和AMP945对小鼠乳腺癌4T1细胞体外免疫原性细胞死亡靶点诱导的研究
实验材料：
1)本实验所用药物
Almonertinib由上海MCE提供,Lot No.:65966
AMP945由MCE提供,Lot No.:143253
2)本实验所用抗体
重组Alexa647荧光Anti-Calreticulin抗体(Abcam，Cat No.:ab196159,Lot No.:CR33676773)。Annexin V-调亡检测试剂盒(Beyotime,Cat No.:C1062L,Lot No.:122221220706)。
实验方法：
4T1细胞(来源南京科佰生物科技有限公司，货号：CBP60352)用RPMI 1640(上海元培,Cat No.:L210KJ,Lot No.:F210916)+10％FBS(Gibco,Cat No.:10099-141c,Lot No.:2158737cp)培养，培养条件为37℃、5％CO2。一周两到三次用胰蛋白酶进行常规消化传代处理。当细胞处于指数生长期，贴壁汇合至80％-90％时，收取细胞并进行铺板。4T1细胞用胰蛋白酶进行消化后收取细胞并进行计数，根据计数结果，用RPMI1640+10％FBS对细胞进行稀释，稀释浓度为5万个细胞每毫升，然后进行12孔细胞培养板铺板，每孔铺2ml细胞悬液即10万个细胞。铺板完成后，将细胞放于37℃、5％CO2培养箱中培养。细胞铺展24小时后，设立六组，第一组为对照组，添加培养基，第二组为AMP945，浓度为3μM，第三组为AMP945，浓度为6μM，第四组为Almonertinib，浓度为2μM，第五组为AMP945(3μM)与Almonertinib(2μM)联用，第六组为AMP945(6μM)与Almonertinib(2μM)联用。将药物混合，并在37℃下在5％CO2培养箱中培养48小时。
实验结果
药物作用48h后，收集细胞进行流动分析，并使用流动缓冲液(PBS+2％FBS)和0.5μl AF647清洗细胞两次，每个孔中加入抗钙网蛋白抗体(abcam)，混合。在4℃且避光的条件下孵育，孵育20min后加入流动缓冲液，使用膜联蛋白染色试剂盒(Beyotime)，加入195μl Annexin-V-FITC结合物，与细胞进行吹打混合后，加入5μl Annexin-V-FITC抗体，轻轻混合，最后加入10μl PI染料混合，室温避光孵育15min，将样品送至流式细胞仪上进行信号测定。流式细胞仪分析结果显示两药合用组的CRT阳性率和Annexin-V阳性率显著优于单药组及对照组，见图19a和图19b。
通过引用将本发明中所提及的所有参考文献均完整合并入本文，就如同每一篇文献均单独列出一样。应理解，在阅读了本发明的公开内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落入本申请所附权利要求书所限定的范围内。

Claims (42)

  1. FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂在制备用于在对象中治疗肿瘤的药物中的用途。
  2. FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂的药物组合产品，其用于在对象中治疗肿瘤。
  3. 一种治疗肿瘤的方法，该方法包括向对象施用治疗有效量的FAK抑制剂、表皮生长因子受体酪氨酸激酶抑制剂和免疫检查点抑制剂。
  4. 如权利要求1-3任一项所述的用途、药物组合产品或者方法，其中所述FAK抑制剂和所述表皮生长因子受体酪氨酸激酶抑制剂诱导免疫原性细胞死亡(ICD)。
  5. 如权利要求1-4任一项所述的用途、药物组合产品或者方法，其中所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、APG-2449、AMP945、AMP886或其药学上可接受的盐，优选为IN10018、AMP945、Defactinib、或其药学上可接受的盐，进一步优选为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐，所述IN10018结构如下：
  6. 如权利要求1-5任一项所述的用途、药物组合产品或者方法，其中所述表皮生长因子受体酪氨酸激酶抑制剂为吉非替尼(Gefitinib)、厄洛替尼(Erlotinib)、埃克替尼(Icotinib)、阿法替尼(Afatinib)、达克替尼(Dacomitinib)、克唑替尼(Crizotinib)、Osimertinib(奥希替尼，AZD9291)、阿美替尼(Almonertinib)、艾氟替尼(Alflutinib，又名伏美替尼(Furmonertinib))、EAI045、JBJ-04-125-02、BLU-945、BLU-701、TQB3804、BBT-176、ES-072、BPI-361175、CH7233163或其药学上可接受的盐。
  7. 如权利要求1-6任一项所述的用途、药物组合产品或者方法，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib、Almonertinib、Alflutinib或其药学上可接受的盐。
  8. 如权利要求1-7任一项所述的用途、药物组合产品或者方法，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib或其药学上可接受的盐。
  9. 如权利要求1-8任一项所述的用途、药物组合产品或者方法，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Almonertinib或其药学上可接受的盐。
  10. 如权利要求1-8任一项所述的用途、药物组合产品或者方法，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Alflutinib或其药学上可接受的盐。
  11. 如权利要求1-10任一项所述的用途、药物组合产品或者方法，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体、PD-1/PD-L1小分子抑制剂或者TIGIT抑制剂。
  12. 如权利要求1-11任一项所述的用途、药物组合产品或者方法，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体，进一步的，所述抗PD-1/PD-L1抗体为帕博利珠单抗(pembrolizumab)、替雷利珠单抗(Tislelizumab)、尼伏单抗(Nivolumab)、特瑞普利单抗(Toripalimab)、阿替利珠单抗(Atezolizumab)、度伐单抗(durvalumab)、阿维单抗(Avelumab)、卡瑞利珠单抗(Camrelizumab)、信迪利单抗(Sintilimab)、西米普利单抗(Cemiplimab)、恩沃利单抗(envafolimab)、BMS-936559、JS003、SHR-1316、GS-4224、AN-4005或者MX-10181。
  13. 如权利要求1-11任一项所述的用途、药物组合产品或者方法，其中所述免疫检查点抑制剂为PD-1/PD-L1小分子抑制剂，进一步的，所述PD-1/PD-L1小分子抑制剂为INCB-086550、拉泽替尼(Lazertinib)、IMMH-010、CA-170、ABSK043或者RRx-001。
  14. 如权利要求1-11任一项所述的用途、药物组合产品或者方法，其中所述免疫检查点抑制剂为TIGIT抑制剂，进一步的，所述TIGIT抑制剂为欧司珀利单抗(Ociperlimab/BGB-A1217)、维博利单抗(Vibostolimab)、domvanalimab(AB154)、替瑞利尤单抗(Tiragolumab)、Belrestotug、艾替利单抗(Etigilimab)、ONO-4686、JS-006、AZD-2936、HLX-301、SEA-TGT、M-6223、IBI-939、COM-902、AB-308、AGEN-1777、AK-127、BAT-6021、BAT-6005、ASP-8374、PM-1022、BMS-986207、HB0036或IBI-321。
  15. 如权利要求1-4任一项所述的用途、药物组合产品或者方法，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
  16. 如权利要求1-4任一项所述的用途、药物组合产品或者方法，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Almonertinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
  17. 如权利要求1-4任一项所述的用途、药物组合产品或者方法，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Alflutinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
  18. 如权利要求1-17任一项所述的用途、药物组合产品或者方法，所述FAK抑制剂、所述表皮生长因子受体酪氨酸激酶抑制剂和所述免疫检查点抑制剂被同时或依次施用于所述对象。
  19. 如权利要求1-18任一项所述的用途、药物组合产品或者方法，其中所述肿瘤为膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、神经胶质瘤或者恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)；优选地，所述肿瘤为乳腺癌、卵巢癌、结肠癌(包括结直肠癌)、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤或胰腺癌。
  20. 如权利要求1-19任一项所述的用途、药物组合产品或者方法，其中所述肿瘤为肺癌(包括小细胞肺癌和非小细胞肺癌)或结肠癌(包括结直肠癌)，尤其是非小细胞肺癌或结肠癌(包括结直肠癌)。
  21. 一种试剂盒或药学上可接受的组合物，其包括：
    (a)FAK抑制剂；
    (b)表皮生长因子受体酪氨酸激酶抑制剂；和
    (c)免疫检查点抑制剂。
  22. 如权利要求21所述的试剂盒或组合物，其中所述FAK抑制剂和所述表皮生长因子受体酪氨酸激酶抑制剂诱导免疫原性细胞死亡(ICD)。
  23. 如权利要求21-22任一项所述的试剂盒或组合物，其中所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、APG-2449、AMP945、AMP886或其药学上可接受的盐，优选为IN10018、AMP945、Defactinib、或其药学上可接受的盐，进一步优选为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐，所述IN10018结构如下：
  24. 如权利要求21-23任一项所述的试剂盒或组合物，其中所述表皮生长因子受体酪氨酸激酶抑制剂为吉非替尼(Gefitinib)、厄洛替尼(Erlotinib)、埃克替尼(Icotinib)、阿法替尼(Afatinib)、达克替尼(Dacomitinib)、克唑替尼(Crizotinib)、Osimertinib(奥希替尼，AZD9291)、阿美替尼(Almonertinib)、艾氟替尼(Alflutinib，又名伏美替尼(Furmonertinib))、EAI045、JBJ-04-125-02、BLU-945、BLU-701、TQB3804、BBT-176、ES-072、BPI-361175、CH7233163或其药学上可接受的盐。
  25. 如权利要求21-24任一项所述的试剂盒或组合物，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib、Almonertinib、Alflutinib或其药学上可接受的盐。
  26. 如权利要求21-25任一项所述的试剂盒或组合物，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib或其药学上可接受的盐。
  27. 如权利要求21-25任一项所述的试剂盒或组合物，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Almonertinib或其药学上可接受的盐。
  28. 如权利要求21-25任一项所述的试剂盒或组合物，其中所述表皮生长因子受体酪氨酸激酶抑制剂为Alflutinib或其药学上可接受的盐。
  29. 如权利要求21-28任一项所述的试剂盒或组合物，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体、PD-1/PD-L1小分子抑制剂或者TIGIT抑制剂。
  30. 如权利要求21-29任一项所述的试剂盒或组合物，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体，进一步的，所述抗PD-1/PD-L1抗体为帕博利珠单抗(pembrolizumab)、替雷利珠单抗(Tislelizumab)、尼伏单抗(Nivolumab)、特瑞普利单抗(Toripalimab)、阿替利珠单抗(Atezolizumab)、度伐单抗(durvalumab)、阿维单抗(Avelumab)、卡瑞利珠单抗(Camrelizumab)、信迪利单抗(Sintilimab)、西米普利单抗(Cemiplimab)、恩沃利单抗(envafolimab)、BMS-936559、JS003、SHR-1316、GS-4224、AN-4005或者MX-10181。
  31. 如权利要求21-29任一项所述的试剂盒或组合物，其中所述免疫检查点抑制剂为PD-1/PD-L1小分子抑制剂，进一步的，所述PD-1/PD-L1小分子抑制剂为INCB-086550、拉泽替尼(Lazertinib)、IMMH-010、CA-170、ABSK043或者RRx-001。
  32. 如权利要求21-29任一项所述的试剂盒或组合物，其中所述免疫检查点抑制剂为TIGIT抑制剂，进一步的，所述TIGIT抑制剂为欧司珀利单抗(Ociperlimab/BGB-A1217)、维博利单抗(Vibostolimab)、domvanalimab(AB154)、替瑞利尤单抗(Tiragolumab)、Belrestotug、艾替利单抗(Etigilimab)、ONO-4686、JS-006、AZD-2936、HLX-301、SEA-TGT、M-6223、IBI-939、COM-902、AB-308、AGEN-1777、AK-127、BAT-6021、BAT-6005、ASP-8374、PM-1022、BMS-986207、HB0036或IBI-321。
  33. 如权利要求21-22任一项所述的试剂盒或组合物，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
  34. 如权利要求21-22任一项所述的试剂盒或组合物，其中所述FAK抑 制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Almonertinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
  35. 如权利要求21-22任一项所述的试剂盒或组合物，其中所述FAK抑制剂为IN10018或其药学上可接受的盐，所述表皮生长因子受体酪氨酸激酶抑制剂为Alflutinib或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
  36. 如权利要求21-35任一项所述的试剂盒或组合物，其用作药物。
  37. 如权利要求21-36任一项所述的试剂盒或组合物，其中所述药物用于肿瘤，所述肿瘤为膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、神经胶质瘤或恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)；优选地，所述肿瘤为乳腺癌、卵巢癌、结肠癌(包括结直肠癌)、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤或胰腺癌。
  38. 如权利要求37所述的试剂盒或组合物，其中所述肿瘤为肺癌(包括小细胞肺癌和非小细胞肺癌)或结肠癌(包括结直肠癌)，尤其是非小细胞肺癌或结肠癌(包括结直肠癌)。
  39. FAK抑制剂，其在治疗肿瘤中用于增强表皮生长因子受体酪氨酸激酶抑制剂诱导的免疫原性细胞死亡。
  40. 如权利要求39所述的FAK抑制剂，其中所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、APG-2449、AMP945、AMP886或其药学上可接受的盐，优选为IN10018、AMP945、Defactinib或其药学上可接受的盐，进一步优选为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐，所述IN10018结构如下：
  41. 如权利要求39或40所述的FAK抑制剂，其中所述表皮生长因子受体酪氨酸激酶抑制剂为吉非替尼(Gefitinib)、厄洛替尼(Erlotinib)、埃克替尼(Icotinib)、阿法替尼(Afatinib)、达克替尼(Dacomitinib)、克唑替尼(Crizotinib)、Osimertinib(奥希替尼，AZD9291)、阿美替尼(Almonertinib)、艾氟替尼(Alflutinib，又名伏美替尼(Furmonertinib))、EAI045、JBJ-04-125-02、BLU-945、BLU-701、TQB3804、BBT-176、ES-072、BPI-361175、CH7233163或其药学上可接受的盐；优选地，所述表皮生长因子受体酪氨酸激酶抑制剂为Osimertinib、Almonertinib、Alflutinib或其药学上可接受的盐。
  42. 如权利要求39-41中任一项所述的FAK抑制剂，其中所述肿瘤为膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、神经胶质瘤或者恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)；优选地，所述肿瘤为乳腺癌、卵巢癌、结肠癌(包括结直肠癌)、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤或胰腺癌。
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