Pharmaceutical combinations and uses of FAK inhibitors and EGFR-TKIs

Abstract

A FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor and an immune checkpoint inhibitor are combined to treat tumors, and a FAK inhibitor and an epidermal growth factor receptor tyrosine kinase inhibitor are combined to treat tumors.

Description

[Technical Field]

[0001] This application claims priority to Chinese Patent Application No. 202211077426.8 filed on September 5, 2022, Chinese Patent Application No. 202211166052.7 filed on September 23, 2022, and Chinese Patent Application No. 202310998195.2 filed on August 8, 2023. The disclosures of the above Chinese patent applications are incorporated herein by reference in their entireties as part of this application.

[0002] The present disclosure is in the field of medicinal chemistry. Specifically, the present disclosure relates to the treatment of tumors using focal adhesion kinase (FAK) inhibitors in combination with other drugs. [Background technology]

[0003] Tumors are the second leading cause of death in human health. Immunogenic cell death (ICD) is an additive effect based on programmed cell death. When cancer cells are exposed to chemotherapy or targeted drugs, intracellular stress signals can be activated. These stress signals include endoplasmic reticulum stress (ER stress) and oxidative stress. Under the action of stress signals, cells first attempt to repair the stress. If the damage caused by stress exceeds the cell's repair capacity, the cell initiates a programmed death process. This process is often accompanied by the release of a class of molecules called damage-associated molecular patterns (DAMPs), including calreticulin, annexin A1, and high-mobility group box 1 (HMGB1). These DAMPs are specifically recognized by pattern recognition receptors on antigen-presenting cells (APCs) in the body, inducing their maturation, differentiation, and activation, and then presenting them to immune cells such as effector T cells, thereby generating antigen memory in the immune cells. If tumor cells from the same source are encountered again, the immune cells will specifically recognize and kill them. The new tumor-specific immune response initiated by ICD can increase sensitivity to immune checkpoint inhibitors (ICIs), thereby enhancing the effects of immune checkpoint inhibitors and providing an anti-tumor response with sustained immunological memory.

[0004] FAK, also known as protein tyrosine kinase 2 (PTK2), is a non-receptor tyrosine kinase and a key component of focal adhesion complexes. FAK mediates integrin and growth factor signals and plays an important role in regulating tumor cell invasion, proliferation, and survival.

[0005] EGFR stands for epidermal growth factor receptor. The EGFR gene encodes and produces a receptor protein called the epidermal growth factor receptor. The EGFR protein is a transmembrane protein divided into three parts: one end is located on the outside of the cell, one part is located in the cell membrane, and the other end is located inside the cell. This allows EGFR to bind to other proteins (called ligands) on the outside of the cell, helping the cell receive signals and respond to stimuli. Because receptor-ligand binding is like a key to a lock, both receptors have specific binding "partners." When EGFR binds to a ligand, it attaches to another nearby EGFR to form a complex (dimer), thereby entering an activated state and activating signaling pathways in the cell. EGFR mutations primarily occur in exons 18–21. Among these, deletion mutations in exon 19 and the L858R point mutation in exon 21 are the most common mutation types, accounting for 90% of all mutation types. When a pathogenic genetic mutation occurs in EGFR, the EGFR protein becomes continuously activated, causing the cell to continuously receive proliferation and survival signals, resulting in excessive cell growth and survival (inability to undergo normal apoptosis), and leading to tumor formation.

[0006] Currently, commercially available drugs for EGFR mutations include first-generation targeted drugs, icotinib, gefitinib, and erlotinib, which target mutations in exon 19 and exon 21; second-generation targeted drugs, afatinib, which target mutations in exon 8 and exon 20; and third-generation targeted drugs, osimertinib (also referred to herein as AZD9291), almonertinib, and alflutinib, which target T790M mutations. Targeted drugs for ALK mutations include the first-generation targeted drug crizotinib, the second-generation targeted drugs ceritinib, alectinib, brigutinib, and the third-generation targeted drug lorlatinib. However, drug resistance to these targeted drugs often appears after about one year of drug treatment. Overcoming resistance to targeted drugs or delaying the onset of drug resistance and improving the chances of cure are major goals of drug development. Summary of the Invention [Problem to be solved by the invention]

[0007] Therefore, there remains a need to explore ways to improve the efficacy of single targeted drugs, increase response rates, create conditions for immunotherapy through combination therapy, and further overcome drug resistance to offer the possibility of tumor cure. [Means for solving the problem]

[0008] One 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 manufacture of a medicament for treating a tumor in a subject.

[0009] 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 a tumor in a subject.

[0010] Another aspect of the present disclosure provides a method of treating a tumor, comprising administering to a subject in need thereof therapeutically effective amounts of a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor, and an immune checkpoint inhibitor.

[0011] Another aspect of the present disclosure provides a kit or pharmaceutically acceptable composition comprising (a) a FAK inhibitor, (b) an epidermal growth factor receptor tyrosine kinase inhibitor, and (c) an immune checkpoint inhibitor.

[0012] Another aspect of the present disclosure provides the use of a FAK inhibitor and an epidermal growth factor receptor tyrosine kinase inhibitor in the manufacture of a medicament for treating a tumor, wherein the FAK inhibitor is used to enhance immunogenic cell death induced by the epidermal growth factor receptor tyrosine kinase inhibitor.

[0013] Another aspect of the present disclosure provides a FAK inhibitor for use in enhancing immunogenic cell death induced by epidermal growth factor receptor tyrosine kinase inhibitors in the treatment of tumors.

[0014] Another aspect of the present disclosure provides a method for treating a tumor, comprising administering to a subject in need thereof therapeutically effective amounts of a FAK inhibitor and an epidermal growth factor receptor tyrosine kinase inhibitor, wherein the FAK inhibitor is used to enhance immunogenic cell death induced by the epidermal growth factor receptor tyrosine kinase inhibitor.

[0015] 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 manufacture of a medicament for the combined treatment of a tumor.

[0016] Another aspect of the present disclosure provides use of a FAK inhibitor in the manufacture of a combination medicament with an epidermal growth factor receptor tyrosine kinase inhibitor and an immune checkpoint inhibitor for treating a tumor.

[0017] Another aspect of the present disclosure provides the use of an epidermal growth factor receptor tyrosine kinase inhibitor in the manufacture of a combination medicament with a FAK inhibitor and an immune checkpoint inhibitor for treating a tumor.

[0018] Another aspect of the present disclosure provides the use of an immune checkpoint inhibitor in the manufacture of a combination medicament with a FAK inhibitor and an epidermal growth factor receptor tyrosine kinase inhibitor for treating a tumor.

[0019] Another aspect of the present disclosure provides the use of a FAK inhibitor in the manufacture of a medicament for the combined treatment of a tumor with an epidermal growth factor receptor tyrosine kinase inhibitor and an immune checkpoint inhibitor.

[0020] Another aspect of the present disclosure provides the use of an epidermal growth factor receptor tyrosine kinase inhibitor in the manufacture of a medicament for the combined treatment of a tumor with a FAK inhibitor and an immune checkpoint inhibitor.

[0021] Another aspect of the present disclosure provides the use of an immune checkpoint inhibitor in the manufacture of a medicament for the combined treatment of a tumor with a FAK inhibitor and an epidermal growth factor receptor tyrosine kinase inhibitor.

[0022] Another aspect of the present disclosure provides a kit comprising a FAK inhibitor and instructions indicating that the FAK inhibitor can be used in combination treatment of a tumor with an epidermal growth factor receptor tyrosine kinase inhibitor and an immune checkpoint inhibitor.

[0023] Another aspect of the present disclosure provides a kit comprising an epidermal growth factor receptor tyrosine kinase inhibitor and instructions indicating that the epidermal growth factor receptor tyrosine kinase inhibitor can be used in a combination treatment of a tumor with a FAK inhibitor and an immune checkpoint inhibitor.

[0024] Another aspect of the present disclosure provides a kit comprising an immune checkpoint inhibitor and instructions indicating that the immune checkpoint inhibitor can be used in a combination treatment of a tumor with a FAK inhibitor and an epidermal growth factor receptor tyrosine kinase inhibitor.

[0025] Another aspect of the present disclosure provides a method of treating a tumor, comprising administering to a subject in need thereof therapeutically effective amounts of a FAK inhibitor and an epidermal growth factor receptor tyrosine kinase inhibitor.

[0026] 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 a tumor in a subject in need thereof.

[0027] Another aspect of the present disclosure provides the use of a FAK inhibitor and an epidermal growth factor receptor tyrosine kinase inhibitor in the manufacture of a pharmaceutical combination for treating a tumor.

[0028] Another aspect of the present disclosure provides the use of a FAK inhibitor in the manufacture of a combination medicament with an epidermal growth factor receptor tyrosine kinase inhibitor for treating a tumor.

[0029] Another aspect of the present disclosure provides the use of an epidermal growth factor receptor tyrosine kinase inhibitor in the manufacture of a combination medicament with a FAK inhibitor for treating a tumor.

[0030] Another aspect of the present disclosure provides the use of a FAK inhibitor and an epidermal growth factor receptor tyrosine kinase inhibitor in the manufacture of a medicament for the combined treatment of a tumor.

[0031] Another aspect of the present disclosure provides the use of a FAK inhibitor in the manufacture of a medicament for the combination treatment of a tumor with an epidermal growth factor receptor tyrosine kinase inhibitor.

[0032] Another aspect of the present disclosure provides the use of an epidermal growth factor receptor tyrosine kinase inhibitor in the manufacture of a medicament for the combination treatment of a tumor with a FAK inhibitor.

[0033] Another aspect of the present disclosure provides a kit comprising a FAK inhibitor and instructions indicating that the FAK inhibitor can be used in combination treatment of a tumor with an epidermal growth factor receptor tyrosine kinase inhibitor.

[0034] Another aspect of the present disclosure provides a kit comprising an epidermal growth factor receptor tyrosine kinase inhibitor and instructions indicating that the epidermal growth factor receptor tyrosine kinase inhibitor can be used in combination treatment of a tumor with a FAK inhibitor.

[0035] In some cases, the FAK inhibitor is IN10018, defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886, or a pharmaceutically acceptable salt thereof; alternatively, the FAK inhibitor is IN10018, AMP945, defactinib, or a pharmaceutically acceptable salt thereof; and further alternatively, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; or alternatively, the FAK inhibitor is the tartrate salt of IN10018, the structure of which is as follows:

[0036] [ka]

[0037] Defactinib, also known as difatini, has a CAS number of 1345713-71-4, GSK2256098 has a CAS number of 1224887-10-8, PF-00562271 has a CAS number of 717907-75-0, VS-4718 has a CAS number of 1061353-68-1, APG-2449 was developed by Ascentage Pharma, and AMP945 has a CAS number of 1393653-34-3.

[0038] Optionally, the epidermal growth factor receptor tyrosine kinase inhibitor is gefitinib, erlotinib, icotinib, afatinib, dacomitinib, crizotinib, osimertinib (AZD9291), almonertinib, alflutinib (also known as flumonertinib), EAI045, JBJ-04-125-02, BLU-945, BLU-701, TQB3804, BBT-176, ES-072, BPI-361175, CH7233163, or a pharmaceutically acceptable salt thereof.

[0039] The CAS number for gefitinib is 184475-35-2, the CAS number for erlotinib is 183321-74-6, the CAS number for icotinib is 610798-31-7, the CAS number for afatinib is 850140-72-6, the CAS number for crizotinib is 877399-52-5, the CAS number for osimertinib (AZD9291) is 1421373-65-0, and the CAS number for Almonel is 1421373-65-0. The CAS number for alflutinib is 1899921-05-1, the CAS number for alflutinib (also known as flumonertinib) is 1869057-83-9, the CAS number for EAI045 is 1942114-09-1, the CAS number for JBJ-04-125-02 is 2060610-53-7, the CAS number for BLU-945 is 2660250-10-0, and the CAS number for BLU-701 is Blueprint It was jointly developed by Medicines Corp and Zai Lab, TQB3804's CAS number is 2267329-76-8, BBT-176 was developed by Bridge Biotherapeutics, Inc., ES-072 was developed by Bosheng Pharmaceutical Co Ltd., BPI-361175 was developed by Betta Pharmaceuticals, and CH7233163 was developed by Chugai Pharmaceutical Co., Ltd.

[0040] Optionally, the epidermal growth factor receptor tyrosine kinase inhibitor is osimertinib, almonertinib, alfurtiniib, or a pharmaceutically acceptable salt thereof.

[0041] Optionally, the epidermal growth factor receptor tyrosine kinase inhibitor is osimertinib or a pharmaceutically acceptable salt thereof.

[0042] Optionally, the epidermal growth factor receptor tyrosine kinase inhibitor is almonertinib or a pharmaceutically acceptable salt thereof.

[0043] Optionally, the epidermal growth factor receptor tyrosine kinase inhibitor is alflutinib or a pharmaceutically acceptable salt thereof.

[0044] Optionally, the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, a PD-1 / PD-L1 small molecule inhibitor, or a TIGIT inhibitor.

[0045] Optionally, the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, and further, the anti-PD-1 / PD-L1 antibody is pembrolizumab, tislelizumab, nivolumab, toripalimab, atezolizumab, durvalumab, avelumab, camrelizumab, sintilimab, cemiplimab, embafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005, or MX-10181.

[0046] Optionally, the immune checkpoint inhibitor is a PD-1 / PD-L1 small molecule inhibitor, and further wherein the PD-1 / PD-L1 small molecule inhibitor is INCB-086550, lazertinib, IMMH-010, CA-170, ABSK043, or RRx-001.

[0047] In some cases, the immune checkpoint inhibitor is a TIGIT inhibitor, and further the TIGIT inhibitor is osipeliumab (BGB-A1217), vibostolimab, domvanalimab (AB154), tiragolumab, berrestug, 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.

[0048] Optionally, 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor.

[0049] Optionally, 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0050] Optionally, 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor.

[0051] Optionally, 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0052] Optionally, 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor.

[0053] Optionally, 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0054] Optionally, the FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.

[0055] In some cases, 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), stomach cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, smooth muscle sarcoma, liposarcoma, nasopharyngeal carcinoma, neuroendocrine carcinoma, ovarian cancer, salivary gland cancer, metastasis from spindle cell carcinoma, anaplastic large cell lymphoma, anaplastic thyroid carcinoma, non-Hodgkin's lymphoma, Hodgkin's lymphoma, glioma or hematological malignancies such as acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), chronic lymphocytic leukemia (CLL) and chronic myeloid leukemia (CML).

[0056] Optionally, the tumor is instead 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.

[0057] Optionally, the tumor is lung cancer (including small cell lung cancer and non-small cell lung cancer) or colon cancer (including colorectal cancer).

[0058] Optionally, the tumor is non-small cell lung cancer or colon cancer (including colorectal cancer).

[0059] In order to more clearly describe the technical solutions of the examples of the present disclosure, the drawings of the examples are briefly introduced below. Obviously, the drawings in the following description only relate to some examples of the present disclosure and are not intended to limit the present disclosure. [Brief explanation of the drawings]

[0060] [Figure 1] FIG. 1 shows white light micrographs of lung cancer KPL cells after 48 hours of incubation with a drug in Example 1. [Figure 2] Figure 2a is a graph showing the percentage of CRT-positive KPL cells after incubating lung cancer KPL cells with a drug for 48 hours in Example 1. Figure 2b is a graph showing the percentage of Annexin V-positive KPL cells after incubating lung cancer KPL cells with a drug for 48 hours in Example 1. [Figure 3] 1 shows white light micrographs of lung cancer KPL cells after 48 hours of incubation with the drug in Example 2. FIG. [Figure 4] Figure 4a is a graph showing the percentage of CRT-positive KPL cells after incubating lung cancer KPL cells with a drug for 48 hours in Example 2. Figure 4b is a graph showing the percentage of Annexin V-positive KPL cells after incubating lung cancer KPL cells with a drug for 48 hours in Example 2. [Figure 5] 1 shows white light micrographs of lung cancer KPL cells after 48 hours of incubation with drugs in Example 3. FIG. [Figure 6]Figure 6a is a graph showing the percentage of CRT-positive KPL cells after incubating lung cancer KPL cells with a drug for 48 hours in Example 3. Figure 6b is a graph showing the percentage of Annexin V-positive KPL cells after incubating lung cancer KPL cells with a drug for 48 hours in Example 3. [Figure 7] FIG. 1 is a graph showing the early and late apoptotic status of cells after FAK silencing in combination with 0.3 nM AZD9291 treatment for 48 hours. [Figure 8] FIG. 10 is a graph showing enhanced release and exposure of calreticulin after FAK silencing combined with 0.3 nM AZD9291 treatment for 48 hours. [Figure 9] 1 is a graph showing the synergistic killing effect of different dosages of AZD9291 in combination with 3 μM and 5 μM IN10018, respectively, on non-small cell lung cancer cells HCC827 for 48 hours. [Figure 10] Figure 1 is a graph showing the detection of cell apoptosis after 48 hours of combined treatment with 0.3 nM AZD9291 and 3 μM IN10018. [Figure 11] FIG. 1 shows that the combination of AZD9291 and IN10018 significantly upregulated endoplasmic reticulum stress in HCC827. [Figure 12] 1 is a graph showing that the combination of AZD9291 and IN10018 enhanced calreticulin release and exposure. [Figure 13] 1 is a graph showing the change in tumor volume after administration of different assay substances in a subcutaneous allograft tumor model of colon cancer CT26 cells in BALB / c mice. [Figure 14] 1 is a graph showing changes in mouse body weight after administration of different assay substances in a subcutaneous allograft tumor model of colon cancer CT26 cells in BALB / c mice. [Figure 15] 1 is a graph showing the change in tumor volume after administration of different assay substances in a subcutaneous allograft tumor model of colon cancer MC38 cells in C57BL / 6 mice. [Figure 16] 1 is a graph showing changes in mouse body weight after administration of different assay substances in a subcutaneous allograft tumor model of colon cancer MC38 cells in C57BL / 6 mice. [Figure 17] Figure 17a is a graph showing the percentage of CRT-positive 4T1 cells after incubating breast cancer 4T1 cells with drugs for 48 hours in Example 8. Figure 17b is a graph showing the percentage of Annexin V-positive 4T1 cells after incubating breast cancer 4T1 cells with drugs for 48 hours in Example 8. [Figure 18] Figure 18a is a graph showing the percentage of CRT-positive 4T1 cells after incubating breast cancer 4T1 cells with drugs for 48 hours in Example 9. Figure 18b is a graph showing the percentage of Annexin V-positive 4T1 cells after incubating breast cancer 4T1 cells with drugs for 48 hours in Example 9. [Figure 19] Figure 19a is a graph showing the percentage of CRT-positive 4T1 cells after incubating breast cancer 4T1 cells with drugs for 48 hours in Example 10. Figure 19b is a graph showing the percentage of Annexin V-positive 4T1 cells after incubating breast cancer 4T1 cells with drugs for 48 hours in Example 10. DETAILED DESCRIPTION OF THE INVENTION

[0061] In order to make the objectives, technical solutions and advantages of the examples of the present disclosure more apparent, the technical solutions of the examples of the present disclosure will be clearly and completely described together with the accompanying drawings of the examples of the present disclosure. Obviously, the described examples are only a part of the examples of the present disclosure, but not all of the examples. Based on the described examples of the present disclosure, all other examples obtained by those skilled in the art without creative efforts belong to the scope of protection of the present disclosure.

[0062] The present disclosure may be embodied in other specific forms without departing from the basic attributes of the disclosure. It is understood that any and all embodiments of the present disclosure may be combined with the technical characteristics of any other embodiment or embodiments to yield additional embodiments. The present disclosure includes further embodiments resulting from such combinations.

[0063] All publications and patents mentioned in this disclosure are incorporated herein by reference in their entirety. In the event that the usage or terminology used in any publication or patent incorporated by reference conflicts with the usage or terminology used in this disclosure, the usage and terminology of this disclosure shall control.

[0064] The section headings used in this disclosure are for organizational purposes only and are not to be construed as limitations on the subject matter.

[0065] Unless otherwise defined, all technical and scientific terms used herein have their ordinary meaning within the field to which the claimed subject matter belongs. In the event that there are multiple definitions of a term, those in this disclosure prevail.

[0066] Unless otherwise indicated in the examples provided or otherwise indicated, all numbers of quantitative characteristics, such as dosage amounts, set forth in the specification and claims are understood to be modified in all instances by the term "about." Also, any numerical ranges recited in this application are understood to be intended to include all subranges within the ranges, and any combination of the endpoints of the ranges or subranges.

[0067] As used in this disclosure, "including," "containing," "comprising," and the like mean that the elements appearing before the term encompass the elements listed after the term and their equivalents, but do not exclude elements not listed. As used herein, "containing" or "including" (comprising) can be open, semi-closed, or closed. In other words, the term also includes "consisting essentially of" or "consisting of."

[0068] definition The following terms and symbols used in this application have the following meanings unless otherwise specified in the context: As used herein, the term "FAK inhibitor" refers to an effective inhibitor of FAK, which may be suitable for mammals, particularly humans. In some embodiments, 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:

[0069] [ka]

[0070] Defactinib, also known as Difacini, has a CAS number of 1345713-71-4, GSK2256098 has a CAS number of 1224887-10-8, PF-00562271 has a CAS number of 717907-75-0, VS-4718 has a CAS number of 1061353-68-1, APG-2449 is developed by Ascentage Pharma, and AMP945 has a CAS number of 1393653-34-3.

[0071] In some embodiments, the FAK inhibitor is instead IN10018, AMP945, defactinib or a pharmaceutically acceptable salt thereof, and in some alternative embodiments, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, or the FAK inhibitor is the tartrate salt of IN10018.

[0072] As used herein, the term "epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI)" refers to a drug that selectively and effectively inhibits epidermal growth factor receptor tyrosine kinase. Examples of epidermal growth factor receptor tyrosine kinase inhibitors include, but are not limited to, gefitinib, erlotinib, icotinib, afatinib, dacomitinib, crizotinib, osimertinib (AZD9291), almonertinib, alflutinib (also known as flumonertinib), EAI045, JBJ-04-125-02, BLU-945, BLU-701, TQB3804, BBT-176, ES-072, BPI-361175, CH7233163, and pharmaceutically acceptable salts thereof. In some embodiments, the epidermal growth factor receptor tyrosine kinase inhibitor is osimertinib, almonertinib, alfurtiniib, or a pharmaceutically acceptable salt thereof.

[0073] The CAS number for gefitinib is 184475-35-2, the CAS number for erlotinib is 183321-74-6, the CAS number for icotinib is 610798-31-7, the CAS number for afatinib is 850140-72-6, the CAS number for dacomitinib is 1110813-31-4, the CAS number for crizotinib is 877399-52-5, and the CAS number for osimertinib (AZD9291) is 1421373 -65-0, the CAS number for almonertinib is 1899921-05-1, the CAS number for alflutinib (also known as flumonertinib) is 1869057-83-9, the CAS number for EAI045 is 1942114-09-1, the CAS number for JBJ-04-125-02 is 2060610-53-7, the CAS number for BLU-945 is 2660250-10-0, and the CAS number for BLU-701 is Blueprint It was jointly developed by Medicines Corp and Zai Lab, TQB3804's CAS number is 2267329-76-8, BBT-176 was developed by Bridge Biotherapeutics, Inc., ES-072 was developed by Bosheng Pharmaceutical Co Ltd., BPI-361175 was developed by Betta Pharmaceuticals, and CH7233163 was developed by Chugai Pharmaceutical Co., Ltd.

[0074] As used herein, the term "immune checkpoint inhibitor" refers to a pharmaceutical agent that can improve the activity of the immune system by modulating immune checkpoint pathways (e.g., PD-1, TIGIT, CTLA-4, LAG-3, TIM-3, etc.). In some embodiments, the immune checkpoint inhibitor is a PD-1 / PD-L1 (programmed cell death protein 1) pathway antagonist (also referred to as a "PD-1 inhibitor") or a TIGIT inhibitor. PD-1 inhibitors are also referred to as PD-1 / PD-L1 inhibitors in the present disclosure. For example, in the therapeutic methods, medicaments, and uses of the present disclosure, the PD-1 / PD-L1 inhibitor is an anti-PD-1 / PD-L1 antibody, including, but not limited to, pembrolizumab (kerida / Keytruda), tislelizumab (Baizean), nivolumab, toripalimab (Tuoyi), durvalumab (Imfinzi), avelumab (Bavencio), atezolizumab (MPDL3280A / Tecentriq), BMS-936559 (a fully humanized IgG4 monoclonal antibody against PD-L1), GS-4224, AN-4005, or MX-10181. In some alternative embodiments, the PD-1 inhibitor is toripalimab. In some embodiments, the PD-1 inhibitor is used to treat a human subject. In some embodiments, the 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, or VSIG9) is a receptor of the Ig superfamily and a novel immune checkpoint in addition to PD-1 / PD-L1.For example, in the therapeutic methods, medicaments, and uses disclosed herein, the immune checkpoint inhibitor is a TIGIT inhibitor, including, but not limited to, osipelimab (BGB-A1217), vibostolimab, domvanalimab (AB154), tiragolumab, berlestog, 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. In some embodiments, the TIGIT inhibitor is used to treat a human subject. For the avoidance of ambiguity, all antibodies in this disclosure include bispecific antibodies.

[0075] As used herein, a "pharmaceutical combination" or "pharmaceutical combination product" can refer to a fixed combination in the form of a dosage unit (e.g., all pharmaceutically active ingredients present in one dosage form), or to a product that is administered in combination in a complete kit, as well as a combination of a medication and instructions indicating that the medication can be used in combination with one or more other medications.

[0076] As used herein, "combination therapy" or "combined medication" means that a medication is used in combination with one or more other medications to treat a disease, and includes both combinations of one medication and one or more other medications, as well as combinations of one medication and instructions indicating that the medication can be used in combination with one or more other medications.

[0077] In this application, "simultaneous or sequential administration" refers to simultaneous administration of two or more medicaments, or sequential administration of two or more medicaments at a certain time interval within an administration cycle (e.g., within 4 weeks, 3 weeks, 2 weeks, 1 week, or 24 hours). The mode of administration (e.g., oral administration, intravenous administration, intramuscular administration, or subcutaneous administration) may be the same or different, and the frequency / cycle of administration of the two or more medicaments may be the same or different. When the therapeutic method, product, or use of the present disclosure involves two medicaments, the two medicaments can be administered simultaneously or separately at a certain interval. When the therapeutic method, product, or use of the present disclosure involves three medicaments, the three medicaments can be administered at the same time, or two medicaments can be administered at one time and the remaining one at a different time, or each of the three medicaments can be administered at different times.

[0078] In some embodiments, the PD-1 / PD-L1 inhibitor is administered intravenously (e.g., as an intravenous infusion), subcutaneously, or orally. Alternatively, the PD-1 / PD-L1 inhibitor is administered by intravenous infusion.

[0079] In some embodiments, the TIGIT inhibitor is administered intravenously (e.g., as an intravenous infusion), subcutaneously, or orally. Alternatively, the TIGIT inhibitor is administered by intravenous infusion.

[0080] The ability of immune checkpoint inhibitors to treat cancer depends on the presence of tumor antigen-specific T cells in tumor tissue. This requires that tumor tissue express antigens that distinguish it from its non-transformed counterparts through new protein products called neoantigens. The neoantigen load of a tumor is closely related to its immunogenicity and sensitivity (e.g., sensitivity to checkpoint inhibitor therapy), meaning that poorly immunogenic tumors can be highly resistant to these drugs. Therapies that release tumor antigens that can be digested by APCs, such as those that induce immunogenic cell death (ICD), can promote effective antitumor immunity, especially when combined with checkpoint inhibitors.

[0081] As used herein, the term "treatment" refers to the administration of one or more pharmaceutical agents to a subject suffering from a disease or having symptoms of a disease to cure, alleviate, relieve, alter, treat, ameliorate, or affect the disease or symptoms of the disease. In some embodiments, the disease is a tumor or cancer.

[0082] As used herein, the term "tumor" refers to an abnormal pathological change formed by the abnormal proliferation of clonal cells caused by the loss of normal regulation of cellular growth in local tissues at the genetic level under the action of various tumorigenic factors. Tumors include 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), gastric cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal carcinoma, neuroendocrine carcinoma, ovarian cancer, salivary gland cancer, metastasis from spindle cell carcinoma, anaplastic large cell lymphoma, anaplastic thyroid carcinoma, non-Hodgkin's lymphoma, Hodgkin's lymphoma, glioma, and hematological malignancies. For example, but not limited to, acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), chronic lymphocytic leukemia (CLL), and chronic myelogenous leukemia (CML), in some embodiments the tumor is alternatively 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, in some embodiments the tumor is lung cancer (including small cell lung cancer and non-small cell lung cancer) or colon cancer (including colorectal cancer), in some embodiments the tumor is non-small cell lung cancer or colon cancer (including colorectal cancer).

[0083] As used herein, the term "subject" or "patient" refers to a mammal or a non-mammal. Mammal refers to any member of the mammalian family, including, but not limited to, humans; non-human primates, such as chimpanzees and other ape and monkey species; livestock, such as cows, horses, sheep, goats, and pigs; domestic animals, such as rabbits, dogs, and cats; and laboratory animals, including rodents, such as rats, mice, and guinea pigs. Examples of non-mammals include, but are not limited to, birds, and the like. The term "subject" is not limited to a particular age or sex. In some embodiments, the subject is a human.

[0084] As used herein, the term "pharmaceutically acceptable" means non-toxic, biologically tolerable, and suitable for administration to a subject.

[0085] As used herein, the term "pharmaceutically acceptable salt" includes acid addition salts formed with inorganic acids, such as hydrochlorides, hydrobromides, carbonates, bicarbonates, phosphates, sulfates, sulfites, nitrates, and the like; and acid addition salts formed with organic acids, such as formates, acetates, malates, maleates, fumarates, tartrates, succinates, citrates, lactates, methanesulfonates, p-toluenesulfonates, 2-hydroxyethylsulfonates, benzoates, salicylates, stearates, and salts of the formula HOOC-(CH) n This refers to non-toxic, biologically tolerable acid addition salts suitable for administration to a subject, including, but not limited to, salts of —COOH (where n is 0-4) formed with alkanedicarboxylic acids.

[0086] Additionally, 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. Those skilled in the art can determine, without undue experimentation, various synthetic methods that can be used to prepare non-toxic pharmaceutically acceptable acid addition salts.

[0087] As used herein, the term "pharmaceutically acceptable composition" means that it must be chemically and / or toxicologically compatible with other ingredients included in the preparation and / or compatible with the subject being treated therewith. As used herein, the term "therapeutically effective amount" refers to an amount generally sufficient to produce a beneficial therapeutic effect in a subject. The therapeutically effective amount of the present disclosure can be determined by conventional methods (e.g., modeling, dose escalation studies, or clinical trials) in conjunction with conventional influencing factors (e.g., mode of administration, pharmacokinetics of the compound, severity and course of the disease, medical history of the subject, health status of the subject, degree of response of the subject to the medication, etc.).

[0088] As used herein, the term "inhibition" refers to a reduction in the baseline activity of a biological activity or method.

[0089] As used herein, the term "kit" refers to a box for containing chemical reagents for detecting chemical components, pharmaceutical residues, virus types, etc. The kits of the present disclosure can include (i) one, two, or three of a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor, and an immune checkpoint inhibitor; and (ii) instructions indicating that the FAK inhibitor, the epidermal growth factor receptor tyrosine kinase inhibitor, and the immune checkpoint inhibitor can be used to treat a tumor in a subject. In one embodiment, the kit includes (i) a FAK inhibitor and (ii) instructions indicating that the FAK inhibitor, the epidermal growth factor receptor tyrosine kinase inhibitor, and the immune checkpoint inhibitor can be used to treat a tumor in a subject. In one embodiment, the kit includes (i) an epidermal growth factor receptor tyrosine kinase inhibitor and (ii) instructions indicating that the FAK inhibitor, the epidermal growth factor receptor tyrosine kinase inhibitor, and the immune checkpoint inhibitor can be used to treat a tumor in a subject. In one embodiment, the kit comprises (i) an immune checkpoint inhibitor, and (ii) instructions indicating that a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor, and an immune checkpoint inhibitor can be used to treat a tumor in a subject. In one embodiment, the kit comprises (i) a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor, and an immune checkpoint inhibitor, and (ii) instructions indicating that a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor, and an immune checkpoint inhibitor can be used to treat a tumor in a subject. In one embodiment, the kit comprises (i) a FAK inhibitor, and (ii) instructions indicating that a FAK inhibitor and an epidermal growth factor receptor tyrosine kinase inhibitor can be used to treat a tumor in a subject. In one embodiment, the kit comprises (i) an epidermal growth factor receptor tyrosine kinase inhibitor, and (ii) instructions indicating that an epidermal growth factor receptor tyrosine kinase inhibitor and a FAK inhibitor can be used to treat a tumor in a subject.

[0090] The compounds of the kit may be contained in separate containers. In some cases, two or more compounds are contained in the same container. For example, the kit may include a first container, a second container, a third container, and a package insert, where the first container contains at least one dose of a pharmaceutical agent comprising a FAK inhibitor, the second container contains at least one dose of an epidermal growth factor receptor tyrosine kinase inhibitor, and the third container contains at least one dose of an immune checkpoint inhibitor pharmaceutical agent, and the package insert includes instructions for treating a tumor in a subject with the pharmaceutical agent. The first container, the second container, and the third container may contain the same or different shapes (e.g., vials, syringes, and bottles) and / or materials (e.g., plastic or glass). The kit may also include other materials that can assist in administering the pharmaceutical agent, such as diluents, filters, IV bags and lines, needles, and syringes.

[0091] The exact amount of FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor, or immune checkpoint inhibitor administered to a subject depends on various factors, such as the given drug or compound, pharmaceutical agent, administration route, disease type, symptoms, and identity of the subject or host being treated, but can also be routinely determined by those skilled in the art. For example, determining an effective amount also depends on the degree, severity, and type of cell proliferation. Those skilled in the art can determine the appropriate dosage based on these and other factors.

[0092] The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor can be administered in any suitable manner, for example orally, intravenously, intramuscularly or subcutaneously.

[0093] For example, when administered orally, the medicament can be orally administered with a pharmaceutically acceptable carrier, such as an inert diluent or an absorbable edible carrier. These can be encapsulated in hard or soft shell gelatin capsules, compressed into tablets, or mixed directly with the patient's food. For example, the medicament can be combined with one or more excipients and used in the form of an ingestible tablet, oral tablet, lozenge, capsule, elixir, suspension, syrup, or wafer. Tablets, lozenges, pills, capsules, etc. may further contain a binder such as tragacanth gum, gum arabic, corn starch, or gelatin; an excipient such as dicalcium phosphate; a disintegrant such as corn starch, potato starch, alginic acid, etc.; a lubricant such as magnesium stearate; or a sweetener such as sucrose, fructose, lactose, or aspartame; or a flavoring agent.

[0094] For example, for intravenous or intraperitoneal administration by infusion or injection, pharmaceutical solutions can be prepared in water, optionally mixed with a nontoxic surfactant.

[0095] Exemplary pharmaceutical dosage forms for injection or infusion include sterile aqueous solutions, dispersions, or sterile powders containing the active ingredient and suitable for the extemporaneous preparation of sterile injectable or infusion solutions or dispersions. In any case, the ultimate dosage form should be sterile, fluid, and stable under the conditions of manufacture and storage.

[0096] Sterile injection solutions can be prepared by mixing the required amount of the drug with various other required ingredients as mentioned above in a suitable solvent, and then filtering and sterilizing the mixture. For the sterile powder used to prepare sterile injection solutions, alternative preparation methods can be vacuum drying and freeze-drying techniques, which can produce powders containing the active ingredient plus any other required ingredients present after previous sterile filtration.

[0097] The amount of FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor, or immune checkpoint inhibitor required for treatment may vary depending on the particular reagent selected, as well as the route of administration, the nature of the disease being treated, and the age and condition of the patient, and will ultimately be determined by the attending physician or clinician. However, in general, the dosage may range from about 0.1 to about 50 mg / kg body weight per day.

[0098] The FAK inhibitor is administered to an adult in a dosage range of 5 mg / day to 300 mg / day. In certain embodiments, IN10018 or a pharmaceutically acceptable salt thereof is administered to an adult in a dosage range of 5 mg / day to 100 mg / day. In certain embodiments, IN10018 or a pharmaceutically acceptable salt thereof is administered to an adult in a dosage range of 25 mg / day to 100 mg / day, the dosage being calculated on the free base.

[0099] The epidermal growth factor receptor tyrosine kinase inhibitor is administered to an adult in a dosage range of 2 to 250 mg per day. In a specific embodiment, osimertinib or a pharmaceutically acceptable salt thereof is administered to an adult in a dosage range of 2 to 250 mg, for example, 80 mg, per day, the dosage being calculated based on the osimertinib dose; almonertinib or a pharmaceutically acceptable salt thereof is administered to an adult in a dosage range of 2 to 250 mg, for example, 110 mg, per day, the dosage being calculated based on the almonertinib dose; and alfutinib or a pharmaceutically acceptable salt thereof is administered to an adult in a dosage range of 2 to 250 mg, for example, 80 mg, per day, the dosage being calculated based on the alfutinib dose.

[0100] The immune checkpoint inhibitor is administered to an adult at a dosage of 2 to 10 mg / kg or 50 to 1200 mg every 2 to 3 weeks. In a specific embodiment, the immune checkpoint inhibitor is administered to an adult at a dosage of 3 to 10 mg / kg or 100 to 1200 mg every 2 to 3 weeks.

[0101] All technical and scientific terms used herein that do not have a specific definition have the meaning commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0102] In some embodiments, the present disclosure also discloses:

[0103] 1. A FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor and an immune checkpoint inhibitor for use in a method of treating a tumor in a subject.

[0104] 2. The FAK inhibitor is IN10018, defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886, or a pharmaceutically acceptable salt thereof; alternatively, the FAK inhibitor is IN10018, AMP945, defactinib, or a pharmaceutically acceptable salt thereof; and alternatively, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; or the FAK inhibitor is the tartrate salt of IN10018, and the structure of IN10018 is as follows:

[0105] [ka]

[0106] 2. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to embodiment 1, wherein

[0107] 3. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to embodiment 1 or 2, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is gefitinib, erlotinib, icotinib, afatinib, dacomitinib, crizotinib, osimertinib (AZD9291), almonertinib, alflutinib (also known as flumonertinib), EAI045, JBJ-04-125-02, BLU-945, BLU-701, TQB3804, BBT-176, ES-072, BPI-361175, CH7233163 or a pharmaceutically acceptable salt thereof.

[0108] 4. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 1 to 3, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is osimertinib, almonertinib, alfurtiniib or a pharmaceutically acceptable salt thereof.

[0109] 5. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 1 to 4, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is osimertinib or a pharmaceutically acceptable salt thereof.

[0110] 6. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 1 to 4, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is almonertinib or a pharmaceutically acceptable salt thereof.

[0111] 7. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 1 to 4, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is alflutinib or a pharmaceutically acceptable salt thereof.

[0112] 8. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 1 to 7, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, a PD-1 / PD-L1 small molecule inhibitor or a TIGIT inhibitor.

[0113] 9. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor, and immune checkpoint inhibitor according to any one of embodiments 1 to 8, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, and further wherein the anti-PD-1 / PD-L1 antibody is pembrolizumab, tislelizumab, nivolumab, toripalimab, atezolizumab, durvalumab, avelumab, camrelizumab, sintilimab, cemiplimab, embafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005, or MX-10181.

[0114] 10. The use, pharmaceutical combination product or method according to any one of embodiments 1 to 8, wherein the immune checkpoint inhibitor is a PD-1 / PD-L1 small molecule inhibitor, and further wherein the PD-1 / PD-L1 small molecule inhibitor is INCB-086550, lazertinib, IMMH-010, CA-170, ABSK043 or RRx-001.

[0115] 11. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor, and immune checkpoint inhibitor according to any one of embodiments 1 to 8, wherein the immune checkpoint inhibitor is a TIGIT inhibitor, and further wherein the TIGIT inhibitor is osipeliumab (BGB-A1217), vibostolimab, domvanalimab (AB154), tiragolumab, berlestog, 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.

[0116] 12. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 1 to 4, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0117] 13. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 1 to 4, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0118] 14. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 1 to 4, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0119] 15. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 1 to 14, administered simultaneously or sequentially to a subject.

[0120] 16. 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), gastric cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal carcinoma, neuroendocrine carcinoma, ovarian cancer, salivary gland cancer, metastasis from spindle cell carcinoma, anaplastic large cell lymphoma, anaplastic thyroid cancer, non-homologous thyroid cancer, thyroid cancer, thyroid cancer, ovarian cancer, ovarian cancer, ovarian cancer, thyroid ... 16. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 1 to 15, wherein the tumor is FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor ...

[0121] 17. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 1 to 16, wherein the tumor is lung cancer (including small cell lung cancer and non-small cell lung cancer) or colon cancer (including colorectal cancer), or the tumor is non-small cell lung cancer or colon cancer (including colorectal cancer).

[0122] 18. (a) FAK inhibitors, (b) an epidermal growth factor receptor tyrosine kinase inhibitor, and (c) Immune checkpoint inhibitors 20. A kit or pharmaceutically acceptable composition comprising:

[0123] 19. The FAK inhibitor is IN10018, defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886, or a pharmaceutically acceptable salt thereof; alternatively, the FAK inhibitor is IN10018, AMP945, defactinib, or a pharmaceutically acceptable salt thereof; and further alternatively, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; or the FAK inhibitor is the tartrate salt of IN10018, and the structure of IN10018 is

[0124] [ka]

[0125] 19. The kit or composition of embodiment 18, wherein

[0126] 20. The kit or composition of embodiment 18 or 19, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is gefitinib, erlotinib, icotinib, afatinib, dacomitinib, crizotinib, osimertinib (AZD9291), almonertinib, alflutinib (also known as flumonertinib), EAI045, JBJ-04-125-02, BLU-945, BLU-701, TQB3804, BBT-176, ES-072, BPI-361175, CH7233163, or a pharmaceutically acceptable salt thereof.

[0127] 21. The kit or composition of any one of embodiments 18 to 20, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is osimertinib, almonertinib, alfurtiniib, or a pharmaceutically acceptable salt thereof.

[0128] 22. The kit or composition according to any one of embodiments 18 to 21, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is osimertinib or a pharmaceutically acceptable salt thereof.

[0129] 23. The kit or composition according to any one of embodiments 18 to 21, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is almonertinib or a pharmaceutically acceptable salt thereof.

[0130] 24. The 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.

[0131] 25. The kit or composition of any one of embodiments 18 to 24, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, a PD-1 / PD-L1 small molecule inhibitor, or a TIGIT inhibitor.

[0132] 26. The kit or composition of any one of embodiments 18 to 25, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, and further wherein the anti-PD-1 / PD-L1 antibody is pembrolizumab, tislelizumab, nivolumab, toripalimab, atezolizumab, durvalumab, avelumab, camrelizumab, sintilimab, cemiplimab, embafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005, or MX-10181.

[0133] 27. The use, pharmaceutical combination product or method according to any one of embodiments 18 to 25, wherein the immune checkpoint inhibitor is a PD-1 / PD-L1 small molecule inhibitor, and further wherein the PD-1 / PD-L1 small molecule inhibitor is INCB-086550, lazertinib, IMMH-010, CA-170, ABSK043 or RRx-001.

[0134] 28. The kit or composition of any one of embodiments 18 to 25, wherein the immune checkpoint inhibitor is a TIGIT inhibitor, and further wherein the TIGIT inhibitor is osipeliumab (BGB-A1217), vibostolimab, domvanalimab (AB154), tiragolumab, berlestog, 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.

[0135] 29. The kit or composition of any one of embodiments 18 to 21, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0136] 30. The kit or composition of any one of embodiments 18 to 21, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0137] 31. The kit or composition of any one of embodiments 18 to 21, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0138] 32. The kit or composition according to any one of embodiments 18 to 31, wherein the composition is for pharmaceutical use.

[0139] 33. The medicament is used to treat a tumor, and the tumor is selected from the group consisting of 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), stomach cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal carcinoma, neuroendocrine carcinoma, ovarian cancer, salivary gland cancer, spindle cell carcinoma, and the like. 33. The kit or composition of embodiment 32, wherein the tumor is metastasis from cancer, anaplastic large cell lymphoma, anaplastic thyroid carcinoma, non-Hodgkin's lymphoma, Hodgkin's lymphoma, glioma, or a hematological malignancy such as acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), chronic lymphocytic leukemia (CLL), and chronic myelogenous leukemia (CML), or wherein 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, or pancreatic cancer.

[0140] 34. The kit 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), or the tumor is non-small cell lung cancer or colon cancer (including colorectal cancer).

[0141] 35. A method for treating a tumor in a subject, comprising administering therapeutically effective amounts of a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor, and an immune checkpoint inhibitor.

[0142] 36. The FAK inhibitor is IN10018, defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886, or a pharmaceutically acceptable salt thereof; alternatively, the FAK inhibitor is IN10018, AMP945, defactinib, or a pharmaceutically acceptable salt thereof; and further alternatively, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; or the FAK inhibitor is the tartrate salt of IN10018, and the structure of IN10018 is

[0143] [ka]

[0144] 36. The method of embodiment 35, wherein

[0145] 37. The method of embodiment 35 or 36, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is gefitinib, erlotinib, icotinib, afatinib, dacomitinib, crizotinib, osimertinib (AZD9291), almonertinib, alflutinib (also known as flumonertinib), EAI045, JBJ-04-125-02, BLU-945, BLU-701, TQB3804, BBT-176, ES-072, BPI-361175, CH7233163, or a pharmaceutically acceptable salt thereof.

[0146] 38. The method of any one of embodiments 35-37, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is osimertinib, almonertinib, alfurtiniib, or a pharmaceutically acceptable salt thereof.

[0147] 39. The method of any one of embodiments 35-38, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is osimertinib or a pharmaceutically acceptable salt thereof.

[0148] 40. The method of any one of embodiments 35-38, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is almonertinib or a pharmaceutically acceptable salt thereof.

[0149] 41. The method of any one of embodiments 35-38, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is alflutinib or a pharmaceutically acceptable salt thereof.

[0150] 42. The method of any one of embodiments 35 to 41, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, a PD-1 / PD-L1 small molecule inhibitor, or a TIGIT inhibitor.

[0151] 43. The method of any one of embodiments 35 to 42, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, and further wherein the anti-PD-1 / PD-L1 antibody is pembrolizumab, tislelizumab, nivolumab, toripalimab, atezolizumab, durvalumab, avelumab, camrelizumab, sintilimab, cemiplimab, embafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005, or MX-10181.

[0152] 44. The use, pharmaceutical combination product or method according to any one of embodiments 35 to 42, wherein the immune checkpoint inhibitor is a PD-1 / PD-L1 small molecule inhibitor, and further wherein the PD-1 / PD-L1 small molecule inhibitor is INCB-086550, lazertinib, IMMH-010, CA-170, ABSK043 or RRx-001.

[0153] 45. The method of any one of embodiments 35 to 42, wherein the immune checkpoint inhibitor is a TIGIT inhibitor, and the TIGIT inhibitor is osipeliumab (BGB-A1217), vibostolimab, domvanalimab (AB154), tiragolumab, berlestog, 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.

[0154] 46. ​​The method of any one of embodiments 35-38, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0155] 47. The method of any one of embodiments 35-38, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0156] 48. The method of any one of embodiments 35-38, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0157] 49. The method of any one of embodiments 35 to 48, wherein the FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.

[0158] 50. 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), gastric cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal carcinoma, neuroendocrine carcinoma, ovarian cancer, salivary gland cancer, metastasis from spindle cell carcinoma, undifferentiated large intestine 50. The method of any one of embodiments 35-49, wherein the tumor is selected from the group consisting of B-cell lymphoma, anaplastic thyroid carcinoma, non-Hodgkin's lymphoma, Hodgkin's lymphoma, glioma, and hematological malignancies, such as acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), chronic lymphocytic leukemia (CLL), and chronic myelogenous leukemia (CML), or wherein 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.

[0159] 51. The method of any one of embodiments 35-50, wherein the tumor is lung cancer (including small cell lung cancer and non-small cell lung cancer) or colon cancer (including colorectal cancer), or wherein the tumor is non-small cell lung cancer or colon cancer (including colorectal cancer).

[0160] 52. A FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor and an immune checkpoint inhibitor for use in a method of treating a tumor by increasing immunogenic cell death in a subject.

[0161] 53. The FAK inhibitor is IN10018, defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886, or a pharmaceutically acceptable salt thereof; alternatively, the FAK inhibitor is IN10018, AMP945, defactinib, or a pharmaceutically acceptable salt thereof; and further alternatively, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; or the FAK inhibitor is the tartrate salt of IN10018, and the structure of IN10018 is

[0162] [ka]

[0163] 53. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to embodiment 52, wherein

[0164] 54. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to embodiment 52 or 53, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is gefitinib, erlotinib, icotinib, afatinib, dacomitinib, crizotinib, osimertinib (AZD9291), almonertinib, alflutinib (also known as flumonertinib), EAI045, JBJ-04-125-02, BLU-945, BLU-701, TQB3804, BBT-176, ES-072, BPI-361175, CH7233163 or a pharmaceutically acceptable salt thereof.

[0165] 55. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 52 to 54, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is osimertinib, almonertinib, alfurtiniib or a pharmaceutically acceptable salt thereof.

[0166] 56. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 52 to 55, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is osimertinib or a pharmaceutically acceptable salt thereof.

[0167] 57. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 52 to 55, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is almonertinib or a pharmaceutically acceptable salt thereof.

[0168] 58. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 52 to 55, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is alflutinib or a pharmaceutically acceptable salt thereof.

[0169] 59. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 52 to 58, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, a PD-1 / PD-L1 small molecule inhibitor or a TIGIT inhibitor.

[0170] 60. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor, and immune checkpoint inhibitor according to any one of embodiments 52 to 59, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, and further wherein the anti-PD-1 / PD-L1 antibody is pembrolizumab, tislelizumab, nivolumab, toripalimab, atezolizumab, durvalumab, avelumab, camrelizumab, sintilimab, cemiplimab, embafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005, or MX-10181.

[0171] 61. The use, pharmaceutical combination product or method according to any one of embodiments 52-59, wherein the immune checkpoint inhibitor is a PD-1 / PD-L1 small molecule inhibitor, and further wherein the PD-1 / PD-L1 small molecule inhibitor is INCB-086550, lazertinib, IMMH-010, CA-170, ABSK043 or RRx-001.

[0172] 62. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor, and immune checkpoint inhibitor according to any one of embodiments 52 to 59, wherein the immune checkpoint inhibitor is a TIGIT inhibitor, and further wherein the TIGIT inhibitor is osipeliumab (BGB-A1217), vibostolimab, domvanalimab (AB154), tiragolumab, berlestog, 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.

[0173] 63. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 52 to 55, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0174] 64. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 52 to 55, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0175] 65. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 52 to 55, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0176] 66. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 52 to 65, administered simultaneously or sequentially to a subject.

[0177] 67. 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), gastric cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal carcinoma, neuroendocrine carcinoma, ovarian cancer, salivary gland cancer, metastasis from spindle cell carcinoma, anaplastic large cell lymphoma, anaplastic thyroid cancer, non-Hodgkin's lymphoma 67. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 52 to 66, wherein the tumor is Hodgkin's lymphoma, Hodgkin's lymphoma, glioma or hematological malignancies such as acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), chronic lymphocytic leukemia (CLL) and chronic myelogenous leukemia (CML), or 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 or pancreatic cancer.

[0178] 68. The FAK inhibitor, epidermal growth factor receptor tyrosine kinase inhibitor and immune checkpoint inhibitor according to any one of embodiments 52 to 67, wherein the tumor is lung cancer (including small cell lung cancer and non-small cell lung cancer) or colon cancer (including colorectal cancer), or the tumor is non-small cell lung cancer or colon cancer (including colorectal cancer).

[0179] 69. A method for treating a tumor by increasing immunogenic cell death in a subject, comprising administering to the subject therapeutically effective amounts of a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor, and an immune checkpoint inhibitor.

[0180] 70. The FAK inhibitor is IN10018, defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886, or a pharmaceutically acceptable salt thereof; alternatively, the FAK inhibitor is IN10018, AMP945, defactinib, or a pharmaceutically acceptable salt thereof; and further alternatively, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; or the FAK inhibitor is the tartrate salt of IN10018, and the structure of IN10018 is

[0181] [ka]

[0182] 70. The method of embodiment 69, wherein

[0183] 71. The method of embodiment 69 or 70, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is gefitinib, erlotinib, icotinib, afatinib, dacomitinib, crizotinib, osimertinib (AZD9291), almonertinib, alflutinib (also known as flumonertinib), EAI045, JBJ-04-125-02, BLU-945, BLU-701, TQB3804, BBT-176, ES-072, BPI-361175, CH7233163, or a pharmaceutically acceptable salt thereof.

[0184] 72. The method of any one of embodiments 69-71, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is osimertinib, almonertinib, alfurtiniib, or a pharmaceutically acceptable salt thereof.

[0185] 73. The method of any one of embodiments 69-72, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is osimertinib or a pharmaceutically acceptable salt thereof.

[0186] 74. The method of any one of embodiments 69-72, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is almonertinib or a pharmaceutically acceptable salt thereof.

[0187] 75. The method of any one of embodiments 69-72, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is alflutinib or a pharmaceutically acceptable salt thereof.

[0188] 76. The method of any one of embodiments 69-75, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, a PD-1 / PD-L1 small molecule inhibitor, or a TIGIT inhibitor.

[0189] 77. The method of any one of embodiments 69 to 76, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, and further wherein the anti-PD-1 / PD-L1 antibody is pembrolizumab, tislelizumab, nivolumab, toripalimab, atezolizumab, durvalumab, avelumab, camrelizumab, sintilimab, cemiplimab, embafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005, or MX-10181.

[0190] 78. The use, pharmaceutical combination product or method according to any one of embodiments 69-76, wherein the immune checkpoint inhibitor is a PD-1 / PD-L1 small molecule inhibitor, and further wherein the PD-1 / PD-L1 small molecule inhibitor is INCB-086550, lazertinib, IMMH-010, CA-170, ABSK043 or RRx-001.

[0191] 79. The method of any one of embodiments 69 to 76, wherein the immune checkpoint inhibitor is a TIGIT inhibitor, and further wherein the TIGIT inhibitor is osipeliumab (BGB-A1217), vibostolimab, domvanalimab (AB154), tiragolumab, berlestog, 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.

[0192] 80. The method of any one of embodiments 69 to 72, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0193] 81. The method of any one of embodiments 69 to 72, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0194] 82. The method of any one of embodiments 69-72, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0195] 83. The method of any one of embodiments 69 to 82, wherein the FAK inhibitor, the epidermal growth factor receptor tyrosine kinase inhibitor and the immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.

[0196] 84. 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), gastric cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal carcinoma, neuroendocrine carcinoma, ovarian cancer, salivary gland cancer, metastasis from spindle cell carcinoma, undifferentiated large intestine 84. The method of any one of embodiments 69 to 83, wherein the tumor is selected from the group consisting of B-cell lymphoma, anaplastic thyroid carcinoma, non-Hodgkin's lymphoma, Hodgkin's lymphoma, glioma, and hematological malignancies, such as acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), chronic lymphocytic leukemia (CLL), and chronic myelogenous leukemia (CML), or wherein 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.

[0197] 85. The method of any one of embodiments 69-84, wherein the tumor is lung cancer (including small cell lung cancer and non-small cell lung cancer) or colon cancer (including colorectal cancer), or the tumor is non-small cell lung cancer or colon cancer (including colorectal cancer).

[0198] 86. Use of a FAK inhibitor in the manufacture of a medicament for treating a tumor in a subject, wherein a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor, and an immune checkpoint inhibitor are administered to the subject.

[0199] 87. Use of an epidermal growth factor receptor tyrosine kinase inhibitor in the manufacture of a medicament for treating a tumor in a subject, wherein a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor, and an immune checkpoint inhibitor are administered to the subject.

[0200] 88. Use of an immune checkpoint inhibitor in the manufacture of a medicament for treating a tumor in a subject, wherein a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor and an immune checkpoint inhibitor are administered to the subject.

[0201] 89. Use of a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor and an immune checkpoint inhibitor in the manufacture of a combination pharmaceutical for treating a tumor.

[0202] 90. Use of a FAK inhibitor in the manufacture of a combination medicine with an epidermal growth factor receptor tyrosine kinase inhibitor and an immune checkpoint inhibitor for treating a tumor.

[0203] 91. Use of an epidermal growth factor receptor tyrosine kinase inhibitor in the manufacture of a combination medicament with a FAK inhibitor and an immune checkpoint inhibitor for treating a tumor.

[0204] 92. Use of an immune checkpoint inhibitor in the manufacture of a combination medicament with a FAK inhibitor and an epidermal growth factor receptor tyrosine kinase inhibitor for treating a tumor.

[0205] 93. Use of a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor and an immune checkpoint inhibitor in the manufacture of a medicament for the combined treatment of a tumor.

[0206] 94. Use of a FAK inhibitor in the manufacture of a medicament for the combined treatment of a tumor with an epidermal growth factor receptor tyrosine kinase inhibitor and an immune checkpoint inhibitor.

[0207] 95. Use of an epidermal growth factor receptor tyrosine kinase inhibitor in the manufacture of a medicament for the combined treatment of a tumor with a FAK inhibitor and an immune checkpoint inhibitor.

[0208] 96. Use of an immune checkpoint inhibitor in the manufacture of a medicament for the combined treatment of a tumor with a FAK inhibitor and an immune checkpoint inhibitor epidermal growth factor receptor tyrosine kinase inhibitor.

[0209] 97. The FAK inhibitor is IN10018, defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886, or a pharmaceutically acceptable salt thereof; alternatively, the FAK inhibitor is IN10018, AMP945, defactinib, or a pharmaceutically acceptable salt thereof; and further alternatively, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; or the FAK inhibitor is a tartrate salt of IN10018, and the structure of IN10018 is as follows:

[0210] [ka]

[0211] The use according to any one of embodiments 86 to 96, wherein

[0212] 98. The use according to any one of embodiments 86 to 97, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is gefitinib, erlotinib, icotinib, afatinib, dacomitinib, crizotinib, osimertinib (AZD9291), almonertinib, alflutinib (also known as flumonertinib), EAI045, JBJ-04-125-02, BLU-945, BLU-701, TQB3804, BBT-176, ES-072, BPI-361175, CH7233163 or a pharmaceutically acceptable salt thereof.

[0213] 99. The use according to any one of embodiments 86 to 98, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is osimertinib, almonertinib, alfurtiniib, or a pharmaceutically acceptable salt thereof.

[0214] 100. The use according to any one of embodiments 86 to 99, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is osimertinib or a pharmaceutically acceptable salt thereof.

[0215] 101. The use according to any one of embodiments 86 to 99, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is almonertinib or a pharmaceutically acceptable salt thereof.

[0216] 102. The use according to any one of embodiments 86 to 99, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is alflutinib or a pharmaceutically acceptable salt thereof.

[0217] 103. The use according to any one of embodiments 86 to 102, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, a PD-1 / PD-L1 small molecule inhibitor or a TIGIT inhibitor.

[0218] 104. The use of any one of embodiments 86 to 103, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, and further wherein the anti-PD-1 / PD-L1 antibody is pembrolizumab, tislelizumab, nivolumab, toripalimab, atezolizumab, durvalumab, avelumab, camrelizumab, sintilimab, cemiplimab, embafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005, or MX-10181.

[0219] 105. The use, pharmaceutical combination product or method according to any one of embodiments 86 to 103, wherein the immune checkpoint inhibitor is a PD-1 / PD-L1 small molecule inhibitor, and further wherein the PD-1 / PD-L1 small molecule inhibitor is INCB-086550, lazertinib, IMMH-010, CA-170, ABSK043 or RRx-001.

[0220] 106. The use according to any one of embodiments 86 to 103, wherein the immune checkpoint inhibitor is a TIGIT inhibitor, and further wherein the TIGIT inhibitor is osipeliumab (BGB-A1217), vibostolimab, domvanalimab (AB154), tiragolumab, berlestog, 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.

[0221] 107. The use according to any one of embodiments 86 to 99, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0222] 108. The use according to any one of embodiments 86 to 99, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0223] 109. The use of any one of embodiments 86 to 99, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0224] 110. The use according to any one of embodiments 86 to 109, wherein the FAK inhibitor, the epidermal growth factor receptor tyrosine kinase inhibitor and the immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.

[0225] 111. 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), gastric cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal carcinoma, neuroendocrine carcinoma, ovarian cancer, salivary gland cancer, metastasis from spindle cell carcinoma, undifferentiated large intestine Use according to any one of embodiments 86 to 110, wherein the tumor is selected from the group consisting of B-cell lymphoma, anaplastic thyroid carcinoma, non-Hodgkin's lymphoma, Hodgkin's lymphoma, glioma or hematological malignancies such as acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), chronic lymphocytic leukemia (CLL) and chronic myelogenous leukemia (CML), or wherein 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 or pancreatic cancer.

[0226] 112. The use according to any one of embodiments 86 to 111, wherein the tumor is lung cancer (including small cell lung cancer and non-small cell lung cancer) or colon cancer (including colorectal cancer), or the tumor is non-small cell lung cancer or colon cancer (including colorectal cancer). [Example]

[0227] The following examples are provided to further illustrate the present disclosure. It should be understood that these examples are used only to illustrate the present disclosure and are not intended to limit the scope of the present disclosure.

[0228] The assay methods in the following examples without specific conditions can be carried out according to conventional conditions for this type of reaction or according to conditions suggested by the manufacturer. All assay materials or reagents used in the following examples are commercially available unless otherwise specified.

[0229] The abbreviations used in the examples have the following meanings:

[0230] [Table 1]

[0231] Example 1 Testing of IN10018 and AZD9291 in lung cancer KPL cells KPL cells (Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences) were cultured in RPMI 1640 (Shanghai Basalmedia, Catalog No. L210KJ, Lot No. F210916) + 10% FBS (Gibco, Catalog No. 10099-141c, Lot No. 2158737cp) and passaged twice. When the cells were in good condition, the culture solution was placed in a 24-well plate. 24 hours after plating, four groups were set up: the first group was a negative control group, to which culture medium was added; the second group was IN10018 at a concentration of 10 μM; the third group was AZD9291 at a concentration of 10 μM; and the fourth group was a combination group of IN10018 and AZD9291, containing IN10018 (10 μM) and AZD9291 (10 μM), respectively. The drugs were mixed and the mixture was incubated at 37°C in a 5% CO2 incubator for 48 hours.

[0232] After 48 hours of drug action, the cells were observed under a microscope, and photographs were taken and stored. The cells were then collected for flow analysis and washed twice with flow buffer (PBS + 2% FBS). 0.5 μl of CoraLite® 488-conjugated GRP94 polyclonal antibody (Proteintech, Cat. No. CL488-14700, Lot No.: 21006579) was added to each well and mixed. The mixture was incubated at 4°C in the dark for 20 minutes. After 20 minutes, the cells were washed twice with flow buffer (PBS + 2% FBS). The cells were then analyzed using a flow cytometer.

[0233] The cells were observed under a microscope. Cells in the AZD9291 single-drug group were in poor condition, while cells in the two-drug combination group were in the worst condition with more cell death, while cells in the negative control group and IN10018 group were in better condition. Flow cytometry results showed that the CRT-positive rate and Annexin V-positive rate in the two-drug combination group were higher than those in the single-drug groups. For details, see Figure 1, Figure 2a, and Figure 2b.

[0234] Example 2 Testing of IN10018 and almonertinib in lung cancer KPL cells KPL cells (Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences) were cultured in RPMI 1640 (Shanghai Basalmedia, Catalog No. L210KJ, Lot No. F210916) + 10% FBS (Gibco, Catalog No. 10099-141c, Lot No. 2158737cp) and passaged twice. When the cells were in good condition, the culture solution was placed in a 24-well plate. 24 hours after plating, four groups were set up: the first group was a negative control group, to which culture medium was added; the second group was IN10018 at a concentration of 10 μM; the third group was almonertinib at a concentration of 4.7 μM; and the fourth group was a combination group of IN10018 and almonertinib, containing IN10018 (10 μM) and almonertinib (4.7 μM), respectively. The drugs were mixed and the mixture was incubated at 37°C in a 5% CO2 incubator for 48 hours.

[0235] After 48 hours of drug action, the cells were observed under a microscope, and photographs were taken and stored. The cells were then collected for flow analysis and washed twice with flow buffer (PBS + 2% FBS). 0.5 μl of CoraLite® 488-conjugated GRP94 polyclonal antibody (Proteintech, Cat. No. CL488-14700, Lot No.: 21006579) was added to each well and mixed. The mixture was incubated at 4°C in the dark for 20 minutes. After 20 minutes, the cells were washed twice with flow buffer (PBS + 2% FBS). The cells were then analyzed using a flow cytometer.

[0236] The cells were observed under a microscope. Cells in the almonertinib single-drug group were in poor condition, while cells in the two-drug combination group were in the worst condition with more cell death, while cells in the negative control group and IN10018 group were in better condition. Flow cytometry results showed that the CRT positive rate and Annexin V positive rate in the two-drug combination group were higher than those in the single-drug groups. For details, see Figure 3, Figure 4a, and Figure 4b.

[0237] Example 3 Testing of IN10018 and alflutinib in lung cancer KPL cells KPL cells (Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences) were cultured in RPMI 1640 (Shanghai Basalmedia, Catalog No. L210KJ, Lot No. F210916) + 10% FBS (Gibco, Catalog No. 10099-141c, Lot No. 2158737cp) and passaged twice. When the cells were in good condition, the culture solution was placed in a 24-well plate. 24 hours after plating, four groups were set up: the first group was a negative control group, to which culture medium was added; the second group was IN10018 at a concentration of 10 μM; the third group was alflutinib at a concentration of 4.7 μM; and the fourth group was a combination group of IN10018 and alflutinib, containing IN10018 (10 μM) and alflutinib (4.7 μM), respectively. The drugs were mixed and the mixture was incubated at 37°C in a 5% CO2 incubator for 48 hours.

[0238] After 48 hours of drug action, the cells were observed under a microscope, and photographs were taken and stored. The cells were then collected for flow analysis and washed twice with flow buffer (PBS + 2% FBS). 0.5 μl of CoraLite® 488-conjugated GRP94 polyclonal antibody (Proteintech, Cat. No. CL488-14700, Lot No.: 21006579) was added to each well and mixed. The mixture was incubated at 4°C in the dark for 20 minutes. After 20 minutes, the cells were washed twice with flow buffer (PBS + 2% FBS). The cells were then analyzed using a flow cytometer.

[0239] The cells were observed under a microscope. Cells in the alflutinib single-drug group were in poor condition, while cells in the two-drug combination group were in the worst condition with more cell death, while cells in the negative control group and IN10018 group were in better condition. Flow cytometry results showed that the CRT positive rate and Annexin V positive rate in the two-drug combination group were higher than those in the single-drug groups. For details, see Figures 5, 6a, and 6b.

[0240] Example 4 Targeted inhibition of FAK enhances immunogenic cell death in non-small cell lung cancer cells in response to AZD9291 Assay Protocol: 1. FAK silencing combined with AZD9291-promoted apoptosis in lung cancer cells HCC827 In an in vitro assay, siRNA technology was used to reduce the expression level of FAK, which was combined with AZD9291 for 48 hours, and apoptosis in control and FAK-silenced cells was detected using a flow cytometer.

[0241] 2. FAK silencing combined with AZD9291 can effectively promote the ICD effect provided in lung cancer cells HCC827.

[0242] In an in vitro assay, siRNA technology was used to reduce the expression level of FAK, which was combined with AZD9291 for 48 hours, and a flow cytometer was used to detect the expression of calreticulin, a major target of ICD.

[0243] Assay antibodies: recombinant Alexa Fluor® 647 anti-calreticulin antibody (Abcam, ab196159), Annexin V Apoptosis Detection Kit (Invitrogen, A35110).

[0244] FAK siRNAs having the sequences shown in the table below were provided by GenePharma. F:CCUGUAUGCCUAUCAGCUUTT; R:AAGCUGAUAGGCAUACAGGTT.

[0245] Assay equipment: Fluorescence microscope (Olympus U-HGLGPS); chemiluminescence imaging system (BIORAD chemidoc touch).

[0246] Assay Results: 1. FAK silencing in combination with AZD9291 significantly improved apoptosis in HCC827 cells

[0247] HCC827 cells were transfected with control siRNA or FAK siRNA at a final concentration of 50 nM. After 24 hours of transfection, 0.3 nM of AZD9291 was added and the cells were treated for 48 hours. Cells were stained using an Annexin V kit and detected using a flow cytometer. The early and late apoptotic values ​​of the cells were compared with those of the DMSO control group, and Graphpad 8.0 was used to generate graphs.

[0248] As shown in Figure 7, the results showed that compared with the control group, late apoptosis in the FAK-silenced group was significantly enhanced after treatment with AZD9291.

[0249] 2. FAK silencing in combination with AZD9291 significantly upregulated the release and exposure of the ICD target calreticulin.

[0250] HCC827 cells were transfected with control siRNA or FAK siRNA at a final concentration of 50 nM. After 24 hours of transfection, 0.3 nM AZD9291 was added and the cells were treated for 48 hours. Calreticulin antibody was used for fluorescent staining, and the staining results were analyzed by flow cytometry. As shown in Figure 8, statistical results from FlowJo software indicated that calreticulin release and exposure were significantly enhanced after FAK silencing and combination with AZD9291.

[0251] Example 5 Examining the enhancement of immunogenic cell death of non-small cell lung cancer cells in response to AZD9291 by IN10018 Assay Protocol: 1. IN10018 in combination with AZD9291 produced a synergistic tumor-killing effect and induced cell apoptosis.

[0252] An in vitro assay was performed to detect the cell killing curve of AZD9291 in lung cancer cells HCC827, and the IC 50 The cytotoxic effects of different doses of AZD9291 in combination with 3 μM IN10018 and 5 μM IN10018 were investigated, respectively. Simultaneously, Annexin V staining was used to detect the apoptotic response to the drugs used alone and in combination using a flow cytometer.

[0253] 2. IN10018 combined with AZD9291 can effectively promote the ICD effect provided in lung cancer cells HCC827.

[0254] In vitro assays used a combination of IN10018 and AZD9291 for 6 hours to detect activation of the endoplasmic reticulum stress signaling pathway, and a combination of the two drugs for 48 hours to assay the expression of calreticulin, a key target of ICD.

[0255] Assay antibody: Recombinant Alexa Fluor® 647 anti-calreticulin antibody (Abcam, ab196159), FAK antibody (CST, 3285S), phospho-eIF2α (Ser51) (CST, 3398), eif2α (CST, 5324), DDIT3 (HUABIO, ET1703-05), HRP-conjugated alpha-tubulin antibody (Proteintech, HRP-66031), Annexin V Apoptosis Detection Kit (Invitrogen, A35110).

[0256] Assay equipment: Fluorescence microscope (Olympus U-HGLGPS); chemiluminescence imaging system (BIORAD chemidoc touch).

[0257] Assay Results: 1. IN10018 in combination with AZD9291 produced a synergistic tumor killing effect.

[0258] 5,000 HCC827 cells per well were plated in a 96-well cell plate. After 24 hours, a fixed concentration of IN10018 was added in combination with different concentrations of AZD9291. The IN10018 concentrations were 3 μM and 5 μM, and the highest AZD9291 concentration was 1 μM. A total of nine concentrations were established using a 3-fold gradient dilution. After 72 hours of co-incubation of the drugs and cells, 10 μL of CCK8 solution was added to each well, and the plate was incubated in an incubator at 37°C and 5% carbon dioxide for 2 hours. A microplate reader was then used to read the data at a set absorbance wavelength of 450 nm. The readings were compared with the DMSO control, and Graphpad 8.0 was used to generate graphs. As shown in Figure 9, the results showed that different dosages of AZD9291 combined with 3 μM and 5 μM IN10018, respectively, for 48 hours had a synergistic killing effect on non-small cell lung cancer cells HCC827.

[0259] 2. IN10018 combined with AZD9291 promoted cell apoptosis

[0260] HCC827 cells were treated with 0.3 nM AZD9291 in combination with 3 μM IN10018. After 48 hours, the cells were harvested, stained using an Annexin V kit, and detected by flow cytometry. The statistical values ​​of early and late apoptosis of cells were compared with those of the DMSO control group, and Graphpad 8.0 was used for graphing. As shown in Figure 10, the results show that early and late apoptosis in the combination group were significantly enhanced compared to the single drug group.

[0261] 3. IN10018 in combination with AZD9291 significantly upregulated endoplasmic reticulum stress in HCC827.

[0262] HCC827 cells were treated with 0.1 nM and 0.3 nM AZD9291 in combination with 3 μM IN10018, respectively. After 6 hours, the protein expression levels of the endoplasmic reticulum stress-related proteins phospho-eIF2α (Ser51) and DDIT3 were assayed. As shown in Figure 11, the results showed that the expression levels of phospho-eIF2α (Ser51) and DDIT3 in the combination group were significantly upregulated compared to the single-drug groups, indicating that the combination of the two drugs significantly upregulated endoplasmic reticulum stress.

[0263] 4. AZD9291 in combination with IN10018 enhanced the release and exposure of ICD-targeted calreticulin.

[0264] HCC827 cells were treated with 0.3 nM AZD9291 in combination with 3 μM IN10018. After 48 hours, calreticulin antibody was used for fluorescent staining, and the staining results were analyzed by flow cytometer. As shown in Figure 12, the statistical results of FlowJo software showed that the release and exposure of calreticulin in the combination group were significantly enhanced compared with those in the single drug group.

[0265] Example 6 Testing the in vivo antitumor efficacy of AZD9291 in a subcutaneous allograft tumor model of colon cancer CT26 cells in BALB / c mice Assay materials: Mice: Six- to eight-week-old female BALB / c mice were purchased from Shanghai Lingchang Biotechnology Co., Ltd. After the animals arrived, they were housed in the assay environment for acclimatization before the assay. Animals were housed in cages (five animals per cage) equipped with an independent air supply system (IVC) in an SPF animal room. All cages, pads, and drinking water were sterilized before use. All experimenters wore protective clothing and latex gloves when performing operations in the animal room. An animal information card for each cage indicated the number of animals in the cage, sex, strain, administration date and time, administration schedule, assay number, group, and date of assay initiation. Cages, food, and drinking water were changed twice weekly. The feeding environment and lighting conditions were as follows: Temperature: 20~26℃; Humidity: 40~70%; Light period: 12 hours light, 12 hours dark; Cage: Made of polycarbonate, 300mm x 180mm x 150mm in volume, padded with corncob, changed twice a week; Food: Assay animals are allowed to eat ad libitum during the entire assay (radiation-sterilized, dry granular food); Drinking water: Assay animals have access to sterile water ad libitum; Animal identification: Assay animals were identified by ear tag.

[0266] The compound information is shown in Table 1.

[0267] [Table 2]

[0268] Colorectal cancer cells CT26 (Nanjing Kebai Biotechnology Co., Ltd., product number: CBP60043) were maintained and passaged by InxMed (Nanjing) Co., Ltd. Cells were cultured in vitro in monolayers in RPMI-1640 medium containing 10% fetal bovine serum in a 37°C, 5% CO2 incubator. Trypsin-EDTA was used for routine digestion and passage two or three times a week. When the cells were in the exponential growth phase and reached 80%-90% saturation, they were harvested, counted, and inoculated.

[0269] Cell seeding and grouping

[0270] 3×10 5 0.1 mL of the cell suspension containing 10 cells was inoculated subcutaneously into the right dorsal region of each mouse. The tumor volume was approximately 63 mm. 3 When tumor volume reached 12 days after cell inoculation, mice were randomly divided into groups according to tumor volume and administered the treatment. Grouping information is shown in Table 2.

[0271] [Table 3]

[0272] Preparation of assay materials See Table 3 for details.

[0273] [Table 4]

[0274] Daily observation of assay animals The formulation and any modifications of this assay protocol were reviewed and approved by the IACUC of ClinBridge Biotech Co. Ltd. The use and welfare of assay animals complied with AAALAC regulations. Animal health and mortality were monitored daily. Routine examinations included observation of tumor growth and the effects of drug treatment on the animals' daily behavior, such as behavioral activity, feeding and drinking (visual inspection only), weight change, appearance signs, or other abnormal conditions. Based on the number of animals in each group, the number of animals that died and any side effects in the group were recorded.

[0275] End of assay If the animal's health continues to deteriorate or the tumor volume exceeds 3,000 mm 3 If the tumor volume exceeds 2000 mm or if the animal is in severe pain or illness, the animal should be euthanized. If any of the following conditions occur, a veterinarian should be contacted and the animal should be euthanized: obvious weakness, weight loss of more than 20%, inability to eat and drink freely, or the average tumor volume in the control group is 2,000 mm 3 The assay is terminated when animals show the following clinical signs and continue to deteriorate: piloerection, kyphosis, pale ears, nose, eyes or paws, shortness of breath, convulsions, continuous diarrhea, dehydration, slowness and vocalization.

[0276] Tumor Measurement and Assay Indicators Tumor diameters were measured three times a week with a vernier caliper. Tumor volume was calculated using the formula V = 0.5 × a × b 2 (where a and b represent the long and short diameters of the tumor, respectively).

[0277] The tumor growth inhibition rate (TGI) (%) was calculated based on the tumor volume on the first day after grouping according to the following formula:

[0278] TGI (%) = [1 - (mean tumor volume of treatment group - mean tumor volume of treatment group at the start of treatment) / (mean tumor volume of vehicle control group - mean tumor volume of vehicle control group at the start of treatment)] x 100%.

[0279] statistical analysis Statistical analysis was performed using Prism Graphpad software based on the tumor volume and tumor mass at the end of the assay. Comparisons between multiple groups were analyzed by two-way analysis of variance, and Fisher's LSD test was used for analysis. P<0.05 was considered to indicate a significant difference.

[0280] Assay Results The in vivo efficacy of the test article AZD9291 and / or PD-L1 antibody in combination with IN10018 was assayed in a subcutaneous allograft tumor model of murine colorectal cancer cell line CT26 in BALB / c mice. After cell inoculation, tumor growth was monitored daily, and animals were divided into groups according to tumor volume 12 days after inoculation. The mean tumor volume of enrolled mice was approximately 63 mm. 3 Due to tumor burden, mice in the control group were euthanized on day 27 post-inoculation, i.e., day 15 post-group administration, terminating all assays.

[0281] On the 15th day after administration to the control group, the tumor volume was 2777.3 ± 705.2 mm 3 The tumor volumes in the AZD9291 + IN10018 (20 + 25 mg / kg), AZD9291 + PDL1 antibody (20 + 10 mg / kg), and AZD9291 + IN10018 + PDL1 antibody (20 + 25 + 10 mg / kg) treatment groups were 1693.4 ± 1207.5 mm, respectively. 3 , 1387.6±859.6mm 3 and 927.2±627.9mm 3Details are shown in Table 4. When comparing the total tumor volume with the control group, the tumor growth inhibition rates (TGI) in the AZD9291 + IN10018 (20 + 25 mg / kg) group, the AZD9291 + PDL1 antibody (20 + 10 mg / kg) group, and the AZD9291 + IN10018 + PDL1 antibody (20 + 25 + 10 mg / kg) group were 40.0% (p<0.0001), 51.2% (p<0.0001), and 62.7% (p<0.0001), respectively. Details are shown in Table 4. Statistical analysis of the total tumor volume compared with that of the AZD9291 + IN10018 + PDL1 antibody (20 + 25 + 10 mg / kg) triple combination group revealed p<0.0001, p=0.0032, and p=0.0745 for the control, AZD9291 + IN10018 (20 + 25 mg / kg), and AZD9291 + PDL1 antibody (20 + 10 mg / kg) treatment groups, respectively. The tumor volumes of each treatment group at different times are shown in Figure 13.

[0282] [Table 5]

[0283] The assay was carried out according to the administration scheme. During the assay, the animals' activities, such as food and water intake, were observed daily, and their body weights were recorded three times a week. The animal weight curves are shown in Figure 14. During the entire administration cycle, the animals in each group did not show significant weight loss and remained in good condition.

[0284] conclusion Compared with the blank control group, each of the AZD9291 + IN10018 (20 + 25 mg / kg), AZD9291 + PDL1 antibody (20 + 10 mg / kg), and AZD9291 + IN10018 + PDL1 antibody (20 + 25 + 10 mg / kg) treatment groups had a significant tumor growth inhibitory effect that was statistically different from the control group. Considering the entire administration cycle, the tumor volume of the AZD9291 + IN10018 + PDL1 antibody (20 + 25 + 10 mg / kg) triple combination group was always smaller than that of the AZD9291 + IN10018 (20 + 25 mg / kg) and AZD9291 + PDL1 antibody (20 + 10 mg / kg) double combination groups, and was statistically different from the AZD9291 + IN10018 (20 + 25 mg / kg) group. Compared with the AZD9291 + IN10018 (20 + 25 mg / kg) and AZD9291 + PDL1 antibody (20 + 10 mg / kg) double-drug combination groups, the triple-drug combination group had a better effect on tumor growth inhibition. At the same time, the animals' body weights changed well, and no abnormalities were observed in their activity, water intake, feeding, and mental state during the entire administration cycle, indicating that the animals tolerated the triple-drug combination of AZD9291 + IN10018 + PDL1 antibody (20 + 25 + 10 mg / kg).

[0285] Example 7 Testing the in vivo antitumor efficacy of AZD9291 in a subcutaneous allograft tumor model of colon cancer MC38 cells in C57BL / 6 mice Assay materials: Mice: Six- to eight-week-old female C57BL / 6 mice were purchased from Shanghai Slack Laboratory Animal Co., Ltd. After the animals arrived, they were housed in the assay environment for acclimatization before the assay. Animals were housed in cages (five animals per cage) equipped with an IVC (independent air supply system) in an SPF animal room. All cages, pads, and drinking water were sterilized before use. All experimenters wore protective clothing and latex gloves when performing operations in the animal room. An animal information card for each cage indicated the number of animals in the cage, sex, strain, administration date and time, administration schedule, assay number, group, and date of assay initiation. Cages, food, and drinking water were changed twice weekly. The feeding environment and lighting conditions were as follows: Temperature: 20~26℃; Humidity: 40~70%; Light period: 12 hours light, 12 hours dark; Cage: Made of polycarbonate, 300mm x 180mm x 150mm in volume, padded with corncob, changed twice a week; Food: Assay animals are allowed to eat ad libitum during the entire assay (radiation-sterilized, dry granular food); Drinking water: Assay animals have access to sterile water ad libitum; Animal identification: Assay animals were identified by ear tag.

[0286] The compound information is shown in Table 5.

[0287] [Table 6]

[0288] Colorectal cancer cells MC38 (Nanjing Kebai Biotechnology Co., Ltd., product number: CBP60825) were maintained and passaged by InxMed (Nanjing) Co., Ltd. Cells were cultured in vitro in monolayers in DMEM medium containing 10% fetal bovine serum in an incubator at 37°C and 5% CO2. Trypsin-EDTA was used for routine digestion and passage two or three times a week. When the cells were in the exponential growth phase and reached 80%-90% saturation, they were harvested, counted, and inoculated.

[0289] Cell seeding and grouping 2×10 5 0.1 mL of the cell suspension containing 10 cells was inoculated subcutaneously into the right dorsal region of each mouse. 3 When tumor volume reached 15 days after cell inoculation, mice were randomly divided into groups according to tumor volume and administered the treatment. Grouping information is shown in Table 6.

[0290] [Table 7]

[0291] Preparation of assay materials See Table 7 for details.

[0292] [Table 8]

[0293] Daily observation of assay animals The formulation and any modifications of this assay protocol were reviewed and approved by the IACUC of ClinBridge Biotech Co. Ltd. The use and welfare of assay animals complied with AAALAC regulations. Animal health and mortality were monitored daily. Routine examinations included observation of the effects of tumor growth and drug treatment on the animals' daily behavior, such as behavioral activity, feeding and drinking (visual inspection only), weight change, external signs or other abnormal conditions. Based on the number of animals in each group, the number of animals that died and any side effects in the group were recorded.

[0294] End of assay If the animal's health continues to deteriorate or the tumor volume exceeds 3,000 mm 3 If the tumor volume exceeds 2000 mm or if the animal is in severe pain or illness, the animal should be euthanized. If any of the following conditions occur, a veterinarian should be contacted and the animal should be euthanized: obvious weakness, weight loss of more than 20%, inability to eat and drink freely, or the average tumor volume in the control group is 2,000 mm 3 The assay is terminated when animals show the following clinical signs and continue to deteriorate: piloerection, kyphosis, pale ears, nose, eyes or paws, shortness of breath, convulsions, continuous diarrhea, dehydration, slowness and vocalization.

[0295] Tumor Measurements and Assay Indicators Tumor diameters were measured three times a week with a vernier caliper. Tumor volume was calculated using the formula V = 0.5 × a × b 2 (where a and b represent the long and short diameters of the tumor, respectively).

[0296] The tumor growth inhibition rate (TGI) (%) was calculated based on the tumor volume on the first day after grouping according to the following formula: TGI (%) = [1 - (mean tumor volume of the treatment group - mean tumor volume of the treatment group at the start of treatment) / (mean tumor volume of the vehicle control group - mean tumor volume of the vehicle control group at the start of treatment)] × 100%.

[0297] statistical analysis Statistical analysis was performed using Prism Graphpad software based on the tumor volume and tumor mass at the end of the assay. Comparisons between multiple groups were analyzed by two-way analysis of variance, and Fisher's LSD test was used for analysis. P<0.05 was considered to indicate a significant difference.

[0298] Assay Results The in vivo efficacy of the test articles AZD9291 and IN10018 in combination with a PD-L1 antibody was assessed in a subcutaneous allograft tumor model of mouse colorectal cancer cell line MC38 in C57BL / 6 mice. After cell inoculation, tumor growth was monitored daily, and 15 days after inoculation, animals were divided into groups according to tumor volume. The mean tumor volume of enrolled mice was approximately 70 mm. 3 Due to tumor burden, mice in the control group were euthanized on day 29 post-inoculation, i.e., day 14 post-group administration, terminating all assays.

[0299] On the 14th day after administration to the control group, the tumor volume was 1984.4±537.8mm 3 The tumor volumes in the PD-L1 antibody (5 mg / kg), AZD9291 (20 mg / kg), IN10018 + PD-L1 antibody (25 + 5 mg / kg), and AZD9291 + IN10018 + PD-L1 antibody (20 + 25 + 5 mg / kg) treatment groups were 1974.8 ± 1629.9 mm, respectively. 3 , 1518.1±728.8mm 3 , 1425.2±889.6mm 3 and 1292.9±934.6mm 3Details are shown in Table 8. When comparing the total tumor volume with the control group, the tumor growth inhibition rates (TGI) in the PD-L1 antibody (5 mg / kg) group, AZD9291 (20 mg / kg) group, IN10018 + PD-L1 antibody (25 + 5 mg / kg) group, and AZD9291 + IN10018 + PD-L1 antibody (20 + 25 + 5 mg / kg) group were 0.5% (p = 0.9728), 24.4% (p = 0.1012), 29.2% (p = 0.0498), and 36.1% (p = 0.0205), respectively. Details are shown in Table 8. Statistical analysis of the total tumor volume compared with the AZD9291 + IN10018 + PD-L1 antibody (20 + 25 + 5 mg / kg) combination group revealed P values ​​of p=0.0205, p=0.0170, p=0.4274, and p=0.6408 for the control group, PD-L1 antibody (5 mg / kg), AZD9291 (20 mg / kg), and IN10018 + PD-L1 antibody (25 + 5 mg / kg) groups, respectively. The tumor volumes of each dosing group at different times are shown in Figure 15.

[0300] [Table 9]

[0301] The assay was performed according to the dosing scheme. During the assay, animal activity, such as food and water intake, was observed daily, and animal weights were recorded three times a week. The animal weight curves are shown in Figure 16. One mouse in the control group died due to tumor rupture 11 days after group administration, and one mouse in the AZD9291 + IN10018 + PD-L1 antibody (20 + 25 + 5 mg / kg) group died due to an error in oral gavage 8 days after group administration. During the entire dosing cycle, animals in the other groups did not show significant weight loss and remained in good condition, indicating that the various dosing methods were well tolerated.

[0302] conclusion Compared to the blank control group, the IN10018 + PD-L1 antibody (25 + 5 mg / kg) and AZD9291 + IN10018 + PD-L1 antibody (20 + 25 + 5 mg / kg) groups each had significant tumor growth inhibitory effects that were statistically different from the control group. Considering the entire administration cycle as a whole, the tumor volume in the AZD9291 + IN10018 + PD-L1 antibody (20 + 25 + 5 mg / kg) group was always smaller than that of each of the other treatment groups, and there was a statistically different effect compared to the PD-L1 antibody (5 mg / kg) single drug group. Although there was no statistical difference between the triple combination group, the AZD9291 (20 mg / kg) group, and the IN10018 + PD-L1 antibody (25 + 5 mg / kg) group, the mean tumor volume in the triple combination group was always smaller than that in each of the other treatment groups, indicating that the AZD9291 + IN10018 + PD-L1 antibody (20 + 25 + 5 mg / kg) triple combination group had a better effect in inhibiting tumor growth. At the same time, except for animals that died naturally due to tumor rupture and human error, the body weight of the other animals in each group changed smoothly, and no abnormalities were observed in their activity, water intake, feeding, or mental state during the entire treatment cycle, indicating that the animals tolerated the triple combination of AZD9291 + IN10018 + PD-L1 antibody (20 + 25 + 5 mg / kg).

[0303] Example 8 Testing AZD9291 and AMP945 for induction of immunogenic cell death targets in murine breast cancer 4T1 cells in vitro Assay materials: 1) Drugs used in this assay AZD9291 was provided by Shanghai Chaolan Chemical Technology Center Co., Ltd., Lot No.: 202012-02.

[0304] AMP945 was provided by MCE, Lot No: 143253.

[0305] 2) Antibodies used in this assay Recombinant Alexa Fluor® 647 anti-calreticulin antibody (Abcam, Catalog No.: ab196159, Lot No.: CR33676773); Annexin V-Apoptosis Detection Kit (Beyotime, Catalog No.: C1062L, Lot No.: 122221220706).

[0306] Assay Method: 4T1 cells (Nanjing Kebai Biotechnology Co., Ltd., product number: CBP60352) were cultured in RPMI 1640 (Shanghai Basalmedia, catalog number: L210KJ, lot number: F210916) + 10% FBS (Gibco, catalog number: 10099-141c, lot number: 2158737cp) at 37°C and 5% CO2. Trypsin was used twice or three times a week for routine digestion and passaging. Cells were harvested and plated when they were in the exponential growth phase and reached 80%-90% confluency. 4T1 cells were digested with trypsin, then harvested and counted. According to the counting results, cells were diluted with RPMI 1640 + 10% FBS to a dilution concentration of 50,000 cells / ml. Next, the cells were plated into 12-well cell culture plates, and 2 ml of cell suspension (100,000 cells) was plated into each well. After plating, the cells were cultured in an incubator at 37°C and 5% CO2. 24 hours after cell plating, six groups were set up: the first group was the control group, to which culture medium was added; the second group was AMP945 at a concentration of 3 μM; the third group was AMP945 at a concentration of 6 μM; the fourth group was AZD9291 at a concentration of 10 μM; the fifth group was AMP945 (3 μM) combined with AZD9291 (10 μM); and the sixth group was AMP945 (6 μM) combined with AZD9291 (10 μM). The drugs were mixed, and the mixture was cultured at 37°C in a 5% CO2 incubator for 48 hours.

[0307] Assay Results After 48 hours of drug action, cells were collected for flow analysis. Cells were washed twice with flow buffer (PBS + 2% FBS) and 0.5 μl of AF647. Anti-calreticulin antibody (Abcam) was added to each well and mixed. The mixture was incubated at 4°C in the dark. After 20 minutes of incubation, flow buffer was added. Using an annexin staining kit (Beyotime), 195 μl of annexin V-FITC conjugate was added and mixed with the cells by pipetting. Next, 5 μl of annexin V-FITC antibody was added and mixed gently. Finally, 10 μl of PI dye was added and mixed. The mixture was incubated at room temperature in the dark for 15 minutes. Samples were sent to a flow cytometer for signal determination. The results of flow cytometry analysis showed that the CRT positive rate and Annexin V positive rate of the two-drug combination group were significantly better than those of the single drug group and the control group, which are shown in Figure 17a and Figure 17b.

[0308] Example 9 Examination of the induction of immunogenic cell death targets in murine breast cancer 4T1 cells in vitro by alflutinib and AMP945 Assay materials: 1) Drugs used in this assay Alflutinib was provided by Shanghai MCE Co., Ltd., Lot No.: 33805.

[0309] AMP945 was provided by MCE, Lot No: 143253.

[0310] 2) Antibodies used in this assay Recombinant Alexa Fluor® 647 anti-calreticulin antibody (Abcam, Catalog No.: ab196159, Lot No.: CR33676773); Annexin V-Apoptosis Detection Kit (Beyotime, Catalog No.: C1062L, Lot No.: 122221220706).

[0311] Assay Method:

[0312] 4T1 cells (Nanjing Kebai Biotechnology Co., Ltd., product number: CBP60352) were cultured in RPMI 1640 (Shanghai Basalmedia, catalog number: L210KJ, lot number: F210916) + 10% FBS (Gibco, catalog number: 10099-141c, lot number: 2158737cp) at 37°C and 5% CO2. Trypsin was used twice or three times a week for routine digestion and passaging. Cells were harvested and plated when they were in the exponential growth phase and reached 80%-90% confluency. 4T1 cells were digested with trypsin, then harvested and counted. According to the counting results, cells were diluted with RPMI 1640 + 10% FBS to a dilution concentration of 50,000 cells / ml. Next, the cells were plated in a 12-well cell culture plate, and 2 ml of the cell suspension (100,000 cells) was plated in each well. After plating, the cells were cultured in an incubator at 37°C and 5% CO2. 24 hours after the cells were plated, six groups were set up: the first group was the control group, to which culture medium was added; the second group was AMP945 at a concentration of 3 μM; the third group was AMP945 at a concentration of 6 μM; the fourth group was alflutinib at a concentration of 10 μM; the fifth group was AMP945 (3 μM) combined with alflutinib (10 μM); and the sixth group was AMP945 (6 μM) combined with alflutinib (10 μM). The drugs were mixed, and the mixture was cultured at 37°C in a 5% CO2 incubator for 48 hours.

[0313] Assay Results After 48 hours of drug action, cells were collected for flow analysis. Cells were washed twice with flow buffer (PBS + 2% FBS) and 0.5 μl of AF647. Anti-calreticulin antibody (Abcam) was added to each well and mixed. The mixture was incubated at 4°C in the dark. After 20 minutes of incubation, flow buffer was added. Using an annexin staining kit (Beyotime), 195 μl of annexin V-FITC conjugate was added and mixed with the cells by pipetting. Next, 5 μl of annexin V-FITC antibody was added and mixed gently. Finally, 10 μl of PI dye was added and mixed. The mixture was incubated at room temperature in the dark for 15 minutes. Samples were sent to a flow cytometer for signal determination. The results of flow cytometry analysis showed that the CRT positive rate and Annexin V positive rate of the two-drug combination group were significantly better than those of the single drug group and the control group, which are shown in Figure 18a and Figure 18b.

[0314] Example 10 Examination of the induction of immunogenic cell death targets in murine breast cancer 4T1 cells in vitro by almonertinib and AMP945 Assay materials: 1) Drugs used in this assay Almonertinib was provided by Shanghai MCE Company, Lot No: 65966.

[0315] AMP945 was provided by MCE, Lot No: 143253.

[0316] 2) Antibodies used in this assay Recombinant Alexa Fluor® 647 anti-calreticulin antibody (Abcam, Catalog No.: ab196159, Lot No.: CR33676773); Annexin V-Apoptosis Detection Kit (Beyotime, Catalog No.: C1062L, Lot No.: 122221220706).

[0317] Assay Method: 4T1 cells (Nanjing Kebai Biotechnology Co., Ltd., product number: CBP60352) were cultured in RPMI 1640 (Shanghai Basalmedia, catalog number: L210KJ, lot number: F210916) + 10% FBS (Gibco, catalog number: 10099-141c, lot number: 2158737cp) at 37°C and 5% CO2. Trypsin was used twice or three times a week for routine digestion and passaging. Cells were harvested and plated when they were in the exponential growth phase and reached 80%-90% confluency. 4T1 cells were digested with trypsin, then harvested and counted. According to the counting results, cells were diluted with RPMI 1640 + 10% FBS to a dilution concentration of 50,000 cells / ml. Next, the cells were plated in a 12-well cell culture plate, and 2 ml of the cell suspension (100,000 cells) was plated in each well. After plating, the cells were cultured in an incubator at 37°C and 5% CO2. 24 hours after the cells were plated, six groups were set up: the first group was the control group, to which culture medium was added; the second group was AMP945 at a concentration of 3 μM; the third group was AMP945 at a concentration of 6 μM; the fourth group was almonertinib at a concentration of 2 μM; the fifth group was AMP945 (3 μM) combined with almonertinib (2 μM); and the sixth group was AMP945 (6 μM) combined with almonertinib (2 μM). The drugs were mixed, and the mixture was cultured at 37°C in a 5% CO2 incubator for 48 hours.

[0318] Assay Results After 48 hours of drug action, cells were collected for flow analysis. Cells were washed twice with flow buffer (PBS + 2% FBS) and 0.5 μl of AF647. Anti-calreticulin antibody (Abcam) was added to each well and mixed. The mixture was incubated at 4°C in the dark. After 20 minutes of incubation, flow buffer was added. Using an annexin staining kit (Beyotime), 195 μl of annexin V-FITC conjugate was added and mixed with the cells by pipetting. Next, 5 μl of annexin V-FITC antibody was added and mixed gently. Finally, 10 μl of PI dye was added and mixed. The mixture was incubated at room temperature in the dark for 15 minutes. Samples were sent to a flow cytometer for signal determination. The results of flow cytometry analysis showed that the CRT positive rate and Annexin V positive rate of the two-drug combination group were significantly better than those of the single drug group and the control group, which are shown in Figure 19a and Figure 19b.

[0319] All references mentioned in this disclosure are incorporated by reference in their entirety as if each document were listed separately. After reading this disclosure, those skilled in the art will be able to make various changes or modifications to the disclosure, and it will be understood that these equivalents also fall within the scope defined by the claims of this application.

Claims

1. Use of a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor, and an immune checkpoint inhibitor in the manufacture of a medicament for treating a tumor in a subject.

2. 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 a tumor in a subject.

3. A method for treating a tumor, comprising administering to a subject therapeutically effective amounts of a FAK inhibitor, an epidermal growth factor receptor tyrosine kinase inhibitor, and an immune checkpoint inhibitor.

4. 4. The use, pharmaceutical combination product or method according to any one of claims 1 to 3, wherein said FAK inhibitor and said epidermal growth factor receptor tyrosine kinase inhibitor induce immunogenic cell death (ICD).

5. The FAK inhibitor is IN10018, defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886, or a pharmaceutically acceptable salt thereof; alternatively, the FAK inhibitor is IN10018, AMP945, defactinib, or a pharmaceutically acceptable salt thereof; or alternatively, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; or alternatively, the FAK inhibitor is a tartrate salt of IN10018, and the structure of IN10018 is 【Chemical 1】 5. The use, pharmaceutical combination product or method according to any one of claims 1 to 4, wherein

6. 6. The use, pharmaceutical combination product or method of any one of claims 1 to 5, wherein said epidermal growth factor receptor tyrosine kinase inhibitor is gefitinib, erlotinib, icotinib, afatinib, dacomitinib, crizotinib, osimertinib (AZD9291), almonertinib, alflutinib (also known as flumonertinib), EAI045, JBJ-04-125-02, BLU-945, BLU-701, TQB3804, BBT-176, ES-072, BPI-361175, CH7233163 or a pharmaceutically acceptable salt thereof.

7. 7. The use, pharmaceutical combination product or method according to any one of claims 1 to 6, wherein said epidermal growth factor receptor tyrosine kinase inhibitor is osimertinib, almonertinib, alfurtiniib or a pharmaceutically acceptable salt thereof.

8. 8. The use, pharmaceutical combination product or method according to any one of claims 1 to 7, wherein said epidermal growth factor receptor tyrosine kinase inhibitor is osimertinib or a pharmaceutically acceptable salt thereof.

9. 9. The use, pharmaceutical combination product or method according to any one of claims 1 to 8, wherein said epidermal growth factor receptor tyrosine kinase inhibitor is almonertinib or a pharmaceutically acceptable salt thereof.

10. 9. The use, pharmaceutical combination product or method according to any one of claims 1 to 8, wherein said epidermal growth factor receptor tyrosine kinase inhibitor is alflutinib or a pharmaceutically acceptable salt thereof.

11. The use, pharmaceutical combination product or method of any one of claims 1 to 10, wherein said immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, a PD-1 / PD-L1 small molecule inhibitor or a TIGIT inhibitor.

12. 12. The use, pharmaceutical combination product or method of any one of claims 1 to 11, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, and further wherein the anti-PD-1 / PD-L1 antibody is pembrolizumab, tislelizumab, nivolumab, toripalimab, atezolizumab, durvalumab, avelumab, camrelizumab, sintilimab, cemiplimab, embafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.

13. 12. The use, pharmaceutical combination product or method of any one of claims 1 to 11, wherein said immune checkpoint inhibitor is a PD-1 / PD-L1 small molecule inhibitor, and further wherein said PD-1 / PD-L1 small molecule inhibitor is INCB-086550, lazertinib, IMMH-010, CA-170, ABSK043 or RRx-001.

14. 12. The use, pharmaceutical combination product or method of any one of claims 1 to 11, wherein said immune checkpoint inhibitor is a TIGIT inhibitor, and further wherein said TIGIT inhibitor is osipeliumab (BGB-A1217), vibostolimab, domvanalimab (AB154), tiragolumab, berlestog, 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.

15. The use, pharmaceutical combination product or method of any one of claims 1 to 4, wherein said FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, said epidermal growth factor receptor tyrosine kinase inhibitor is osimertinib or a pharmaceutically acceptable salt thereof, and said immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or said immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

16. The use, pharmaceutical combination product or method of any one of claims 1 to 4, wherein said FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, said epidermal growth factor receptor tyrosine kinase inhibitor is almonertinib or a pharmaceutically acceptable salt thereof, and said immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or said immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

17. The use, pharmaceutical combination product or method of any one of claims 1 to 4, wherein said FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, said epidermal growth factor receptor tyrosine kinase inhibitor is alflutinib or a pharmaceutically acceptable salt thereof, and said immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or said immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

18. 18. The use, pharmaceutical combination product or method according to any one of claims 1 to 17, wherein the FAK inhibitor, the epidermal growth factor receptor tyrosine kinase inhibitor and the immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.

19. The tumor may be 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), gastric cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal carcinoma, neuroendocrine carcinoma, ovarian cancer, salivary gland cancer, metastasis from spindle cell carcinoma, or anaplastic large cell lymphoma.

19. The use, pharmaceutical combination product or method of any one of claims 1 to 18, wherein the tumor is selected from the group consisting of acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), chronic lymphocytic leukemia (CLL) and chronic myelogenous leukemia (CML), anaplastic thyroid cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma, glioma or hematological malignancies such as acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), chronic lymphocytic leukemia (CLL) and chronic myelogenous leukemia (CML), or wherein 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 or pancreatic cancer.

20. 20. The use, pharmaceutical combination product or method of any one of claims 1 to 19, wherein the tumour is lung cancer (including small cell lung cancer and non-small cell lung cancer) or colon cancer (including colorectal cancer), or wherein the tumour is non-small cell lung cancer or colon cancer (including colorectal cancer).

21. (a) FAK inhibitors, (b) an epidermal growth factor receptor tyrosine kinase inhibitor, and (c) Immune checkpoint inhibitors 20. A kit or pharmaceutically acceptable composition comprising:

22. 22. The kit or composition of claim 21, wherein the FAK inhibitor and the epidermal growth factor receptor tyrosine kinase inhibitor induce immunogenic cell death (ICD).

23. The FAK inhibitor is IN10018, defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886, or a pharmaceutically acceptable salt thereof; alternatively, the FAK inhibitor is IN10018, AMP945, defactinib, or a pharmaceutically acceptable salt thereof; or alternatively, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; or alternatively, the FAK inhibitor is a tartrate salt of IN10018, and the structure of IN10018 is 【Chemistry 2】 23. The kit or composition of claim 21 or 22, wherein:

24. 24. The kit or composition of any one of claims 21 to 23, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is gefitinib, erlotinib, icotinib, afatinib, dacomitinib, crizotinib, osimertinib (AZD9291), almonertinib, alflutinib (also known as flumonertinib), EAI045, JBJ-04-125-02, BLU-945, BLU-701, TQB3804, BBT-176, ES-072, BPI-361175, CH7233163, or a pharmaceutically acceptable salt thereof.

25. The kit or composition of any one of claims 21 to 24, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is osimertinib, almonertinib, alfurtiniib, or a pharmaceutically acceptable salt thereof.

26. 26. The kit or composition of any one of claims 21 to 25, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is osimertinib or a pharmaceutically acceptable salt thereof.

27. The kit or composition according to any one of claims 21 to 25, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is almonertinib or a pharmaceutically acceptable salt thereof.

28. 26. The kit or composition of any one of claims 21 to 25, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is alflutinib or a pharmaceutically acceptable salt thereof.

29. The kit or composition of any one of claims 21 to 28, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, a PD-1 / PD-L1 small molecule inhibitor, or a TIGIT inhibitor.

30. The kit or composition of any one of claims 21 to 29, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, and further wherein the anti-PD-1 / PD-L1 antibody is pembrolizumab, tislelizumab, nivolumab, toripalimab, atezolizumab, durvalumab, avelumab, camrelizumab, sintilimab, cemiplimab, embafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005, or MX-10181.

31. 30. The kit or composition of any one of claims 21 to 29, wherein the immune checkpoint inhibitor is a PD-1 / PD-L1 small molecule inhibitor, and further wherein the PD-1 / PD-L1 small molecule inhibitor is INCB-086550, lazertinib, IMMH-010, CA-170, ABSK043, or RRx-001.

32. The kit or composition of any one of claims 21 to 29, wherein the immune checkpoint inhibitor is a TIGIT inhibitor, and the TIGIT inhibitor is osipeliumab (BGB-A1217), vibostolimab, domvanalimab (AB154), tiragolumab, berlestog, 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.

33. 23. The kit or composition of claim 21 or 22, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

34. 23. The kit or composition of claim 21 or 22, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

35. 23. The kit or composition of claim 21 or 22, wherein 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, and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, or the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

36. A kit or composition according to any one of claims 21 to 35 for use as a medicament.

37. The medicament is used to treat tumors, and the tumor is metastasis from 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), stomach cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal carcinoma, neuroendocrine cancer, ovarian cancer, salivary gland cancer, spindle cell carcinoma, 37. The kit or composition of any one of claims 21 to 36, wherein the tumor is anaplastic large cell lymphoma, anaplastic thyroid carcinoma, non-Hodgkin's lymphoma, Hodgkin's lymphoma, glioma or a hematological malignancy such as acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), chronic lymphocytic leukemia (CLL) and chronic myelogenous leukemia (CML), or 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 or pancreatic cancer.

38. 38. The kit or composition of claim 37, wherein the tumor is lung cancer (including small cell lung cancer and non-small cell lung cancer) or colon cancer (including colorectal cancer), or wherein the tumor is non-small cell lung cancer or colon cancer (including colorectal cancer).

39. A FAK inhibitor for use in enhancing immunogenic cell death induced by epidermal growth factor receptor tyrosine kinase inhibitors in the treatment of tumors.

40. IN10018, defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886, or a pharmaceutically acceptable salt thereof, or IN10018, AMP945, defactinib, or a pharmaceutically acceptable salt thereof, and / or IN10018 or a pharmaceutically acceptable salt thereof, or a tartrate salt of IN10018, wherein the structure of IN10018 is 【Chemistry 3】 The FAK inhibitor of claim 39, wherein:

41. 41. The FAK inhibitor of claim 39 or 40, wherein the epidermal growth factor receptor tyrosine kinase inhibitor is gefitinib, erlotinib, icotinib, afatinib, dacomitinib, crizotinib, osimertinib (AZD9291), almonertinib, alflutinib (also known as flumonertinib), EAI045, JBJ-04-125-02, BLU-945, BLU-701, TQB3804, BBT-176, ES-072, BPI-361175, CH7233163, or a pharmaceutically acceptable salt thereof.

42. The tumor may be 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), gastric cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal carcinoma, neuroendocrine carcinoma, ovarian cancer, salivary gland cancer, metastasis from spindle cell carcinoma, undifferentiated large cell carcinoma 42. The FAK inhibitor of any one of claims 39 to 41, wherein the tumor is lymphoma, anaplastic thyroid cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma, glioma, or hematological malignancy such as acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), chronic lymphocytic leukemia (CLL), and chronic myelogenous leukemia (CML), or 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, or pancreatic cancer.

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