Pharmaceutical composition of FAK inhibitor and topoisomerase inhibitor and application

Abstract

The present disclosure relates to the combination treatment of tumors with FAK inhibitors, topoisomerase inhibitors, and immune checkpoint inhibitors. The present disclosure also relates to the combination treatment of tumors with FAK inhibitors and topoisomerase inhibitors.

Description

Drug combination and use of FAK inhibitor and topoisomerase inhibitor

[0001] This application claims priority to Chinese patent application 202211129639.0 filed on September 16, 2022 and Chinese patent application 202311155482.3 filed on September 7, 2023, and the contents of the above-mentioned Chinese patent application disclosures are hereby incorporated by reference in their entirety as part of this application. Technical Field

[0002] The present invention belongs to the field of medicinal chemistry and specifically relates to the treatment of tumors by combining a focal adhesion kinase (FAK) inhibitor with other drugs. Background Art

[0003] Cancer is the second leading threat to human health. Immunogenic cell death (ICD) is based on the synergistic effects of programmed cell death. When cancer cells are exposed to chemotherapy or targeted therapy drugs, they may activate internal stress signals, including endoplasmic reticulum stress (ER stress) and reactive oxygen species (ROS) stress. Under the influence of stress signals, cells first attempt to repair the stress. If the damage caused by stress exceeds the ability to repair, the cells initiate programmed cell death. This process is often accompanied by the release of a class of molecules called damage-associated molecular patterns (DAMPs). These DAMPs are specifically recognized by pattern recognition receptors on antigen-presenting cells (APCs) in the body, inducing APC maturation, differentiation, and activation. These DAMPs are then progressively presented to immune cells, such as effector T cells, thereby enabling immune cells to develop antigen memory. Upon encountering tumor cells of the same origin, these immune cells will specifically recognize and kill the tumor cells. The new tumor-specific immune response initiated by ICD can increase sensitivity to immune checkpoint inhibitors (ICIs), thereby enhancing the effects of immune checkpoint inhibitors and producing anti-tumor responses with long-lasting immune memory.

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

[0005] The in vivo function of topoisomerases is to release the tension generated during DNA replication and transcription by cleaving DNA chains, removing positive supercoils generated at the front of the replication fork and negative supercoils generated by RNA polymerization during transcription. The enzyme then connects the broken ends of the DNA chains to ensure smooth reaction progression. Topoisomerase inhibitors bind to the corresponding isomerase's readily dissociable complexes, forming a stable complex that blocks the smooth passage of the replication fork, leading to DNA strand breaks and disrupting DNA replication and transcription. These drugs are widely used in cancer treatment. However, they can develop resistance to these drugs over time.

[0006] Chemotherapy, including combinations of topoisomerase inhibitors and immunotherapy, has been widely used in clinical practice. However, there is still room for improvement in efficacy, especially in terms of enhancing efficacy and producing lasting anti-tumor effects.

[0007] Summary of the Invention

[0008] In one aspect, the present disclosure provides uses of a FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor in the preparation of a medicament for treating a tumor in a subject, wherein the topoisomerase inhibitor is not an anthracycline.

[0009] Yet another aspect of the present disclosure provides a pharmaceutical combination product of a FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor for use in treating a tumor in a subject, wherein the topoisomerase inhibitor is not an anthracycline.

[0010] In another aspect, the present disclosure provides a method for treating tumors, comprising administering to a subject in need thereof a therapeutically effective amount of a FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

[0011] In another aspect, the present disclosure provides a kit or a pharmaceutically acceptable composition comprising: (a) a FAK inhibitor; (b) a topoisomerase inhibitor, wherein the topoisomerase inhibitor is not an anthracycline; and (c) an immune checkpoint inhibitor.

[0012] Another aspect of the present disclosure provides use of a FAK inhibitor and a topoisomerase inhibitor in the preparation of a medicament for treating tumors, wherein the FAK inhibitor is used to enhance immunogenic cell death induced by the topoisomerase inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

[0013] Yet another aspect of the present disclosure provides a FAK inhibitor for use in the treatment of tumors to enhance immunogenic cell death induced by a topoisomerase inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

[0014] Another aspect of the present disclosure provides a method for treating tumors, comprising administering to a subject in need thereof a therapeutically effective amount of a FAK inhibitor and a topoisomerase inhibitor, wherein the FAK inhibitor is used to enhance immunogenic cell death induced by the topoisomerase inhibitor, and wherein the topoisomerase inhibitor is not an anthracycline.

[0015] In another aspect, the present disclosure provides use of a FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor in the preparation of a medicament for combined treatment of tumors, wherein the topoisomerase inhibitor is not an anthracycline.

[0016] In another aspect, the present disclosure provides use of a FAK inhibitor in the preparation of a combination medicament for treating tumors with a topoisomerase inhibitor and an immune checkpoint inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

[0017] In another aspect, the present disclosure provides use of a topoisomerase inhibitor in the preparation of a combination drug for treating tumors with a FAK inhibitor and an immune checkpoint inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

[0018] In another aspect, the present disclosure provides use of an immune checkpoint inhibitor in the preparation of a combination medicament for treating tumors with a FAK inhibitor and a topoisomerase inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

[0019] In another aspect, the present disclosure provides use of a FAK inhibitor in the preparation of a medicament for treating tumors in combination with a topoisomerase inhibitor and an immune checkpoint inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

[0020] In another aspect, the present disclosure provides use of a topoisomerase inhibitor in the preparation of a medicament for treating tumors in combination with a FAK inhibitor and an immune checkpoint inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

[0021] In another aspect, the present disclosure provides use of an immune checkpoint inhibitor in the preparation of a medicament for treating tumors in combination with a FAK inhibitor and a topoisomerase inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

[0022] In another aspect, the present disclosure provides a kit comprising: a FAK inhibitor; and instructions, wherein the instructions indicate that the FAK inhibitor can be used in combination with a topoisomerase inhibitor and an immune checkpoint inhibitor to treat tumors, wherein the topoisomerase inhibitor is not an anthracycline.

[0023] In another aspect, the present disclosure provides a kit comprising: a topoisomerase inhibitor; and instructions, wherein the instructions indicate that the topoisomerase inhibitor can be used in combination with a FAK inhibitor and an immune checkpoint inhibitor to treat tumors, wherein the topoisomerase inhibitor is not an anthracycline.

[0024] In another aspect, the present disclosure provides a kit comprising: an immune checkpoint inhibitor; and instructions, wherein the instructions indicate that the immune checkpoint inhibitor can be used in combination with a FAK inhibitor and a topoisomerase inhibitor to treat tumors, wherein the topoisomerase inhibitor is not an anthracycline.

[0025] Another aspect of the present disclosure provides a method for treating tumors, comprising administering to a subject in need thereof a therapeutically effective amount of a FAK inhibitor and a topoisomerase inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

[0026] Yet another aspect of the present disclosure provides a pharmaceutical combination product of a FAK inhibitor and a topoisomerase inhibitor for use in treating a tumor in a subject in need thereof, wherein the topoisomerase inhibitor is not an anthracycline.

[0027] In another aspect, the present disclosure provides use of a FAK inhibitor and a topoisomerase inhibitor in preparing a combined drug for treating tumors, wherein the topoisomerase inhibitor is not an anthracycline.

[0028] In another aspect, the present disclosure provides use of a FAK inhibitor in the preparation of a medicament for use in combination with a topoisomerase inhibitor in treating tumors, wherein the topoisomerase inhibitor is not an anthracycline.

[0029] In another aspect, the present disclosure provides use of a topoisomerase inhibitor in the preparation of a medicament for use in combination with a FAK inhibitor in treating tumors, wherein the topoisomerase inhibitor is not an anthracycline.

[0030] In another aspect, the present disclosure provides use of a FAK inhibitor and a topoisomerase inhibitor in the preparation of a medicament for combined treatment of tumors, wherein the topoisomerase inhibitor is not an anthracycline.

[0031] In another aspect, the present disclosure provides use of a FAK inhibitor in the preparation of a medicament for treating tumors in combination with a topoisomerase inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

[0032] In another aspect, the present disclosure provides use of a topoisomerase inhibitor in the preparation of a medicament for treating tumors in combination with a FAK inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

[0033] In another aspect, the present disclosure provides a kit comprising: a FAK inhibitor; and instructions, wherein the instructions indicate that the FAK inhibitor can be used in combination with a topoisomerase inhibitor to treat tumors, wherein the topoisomerase inhibitor is not an anthracycline.

[0034] In another aspect, the present disclosure provides a kit comprising: a topoisomerase inhibitor; and instructions, wherein the instructions indicate that the topoisomerase inhibitor can be used in combination with a FAK inhibitor to treat tumors, wherein the topoisomerase inhibitor is not an anthracycline.

[0035] Optionally, the FAK inhibitor is IN10018, Defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886, or a pharmaceutically acceptable salt thereof, preferably IN10018, Defactinib, AMP945, or a pharmaceutically acceptable salt thereof, further preferably IN10018 or a pharmaceutically acceptable salt thereof, especially IN10018 tartrate. The structure of IN10018 is as follows:

[0036] Defactinib, also known as defatinib, 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.

[0037] Optionally, the topoisomerase inhibitor is a topoisomerase I inhibitor.

[0038] For example, the topoisomerase I inhibitor is topotecan, irinotecan, belotecan, cositecan, exatecan (DX-8951), indenoisoquinoline, indotecan (LMP-400), indimitecan (LMP-776), simmitecan, gimatecan (ST1481), EC-112002, PLX-038 (NK012), AR-67, or a pharmaceutically acceptable salt thereof.

[0039] The CAS number of the topotecan is 123948-87-8; the CAS number of the irinotecan is 97682-44-5; the CAS number of the belotecan is 256411-32-2; the CAS number of the cositecan is 401905-67-7; the CAS number of the exatecan (DX-8951) is 171335-80-1; the indenoisoquinoline The CAS number of the oquinoline is 97501-75-2; the CAS number of the indotecan (LMP-400) is 915303-09-2; the CAS number of the indimitecan (LMP-776) is 915360-05-3; the CAS number of the simmitecan is 951290-31-6; the CAS number of the gimatecan (ST1481) is 292618-32-7; the EC-112002 was developed by Elucida Oncology Inc., the CAS number of the PLX-038 (NK012) is 86639-52-3; and the CAS number of the AR-67 is 220913-32-6.

[0040] Optionally, the topoisomerase I inhibitor is irinotecan or a pharmaceutically acceptable salt thereof.

[0041] Optionally, the topoisomerase inhibitor is a topoisomerase II inhibitor.

[0042] For example, the topoisomerase II inhibitor is etoposide, teniposide, sabarubicin, or a pharmaceutically acceptable salt thereof.

[0043] The CAS number of the etoposide is 33419-42-0; the CAS number of the teniposide is 29767-20-2; and the CAS number of the sabarubicin is 211100-13-9.

[0044] Optionally, the topoisomerase II inhibitor is etoposide, teniposide, or a pharmaceutically acceptable salt thereof.

[0045] Optionally, the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, a PD-1 / PD-L1 small molecule inhibitor or a TIGIT antibody.

[0046] 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, Atezolizumab, Camrelizumab, Sintilimab, Cemiplimab, envafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.

[0047] Optionally, the immune checkpoint inhibitor is a PD-1 / PD-L1 small molecule inhibitor, and further, the PD-1 / PD-L1 small molecule inhibitor is INCB-086550, Lazertinib, IMH-010, CA-170, ABSK043 or RRx-001.

[0048] Optionally, the immune checkpoint inhibitor is a TIGIT antibody, and further, the TIGIT antibody is Ociperlimab (Ociperlimab / BGB-A1217), Vibostolimab, domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.

[0049] Optionally, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the topoisomerase inhibitor is irinotecan 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.

[0050] Optionally, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the topoisomerase inhibitor is irinotecan or a pharmaceutically acceptable salt thereof; and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0051] Optionally, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the topoisomerase inhibitor is etoposide, teniposide, 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.

[0052] Optionally, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the topoisomerase inhibitor is etoposide, teniposide, or a pharmaceutically acceptable salt thereof; and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0053] Optionally, the FAK inhibitor, the topoisomerase inhibitor and the immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.

[0054] Optionally, 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, bile duct cancer, leiomyosarcoma, Liposarcoma, nasopharyngeal carcinoma, neuroendocrine carcinoma, ovarian cancer, salivary gland cancer, metastases from spindle cell carcinoma, anaplastic large cell lymphoma, anaplastic thyroid cancer, non-Hodgkin lymphoma, Hodgkin 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), chronic myeloid leukemia (CML).

[0055] Optionally, the tumor is breast cancer or colon cancer (including colorectal cancer). BRIEF DESCRIPTION OF THE DRAWINGS

[0056] In order to more clearly illustrate the technical solutions of the embodiments of the present disclosure, the drawings of the embodiments will be briefly introduced below. Obviously, the drawings in the following description only relate to some embodiments of the present disclosure, and are not intended to limit the present invention.

[0057] Figure 1 shows that FAK silencing combined with 2 μM etoposide treatment for 48 hours increased apoptosis in MDA-MB-231 cells.

[0058] FIG2 shows that FAK silencing combined with 2 μM etoposide treatment for 48 hours enhanced the release and exposure of calreticulin.

[0059] FIG3 shows that FAK silencing combined with 2 μM teniposide treatment for 48 hours increased apoptosis in MDA-MB-231 cells.

[0060] FIG4 shows that FAK silencing combined with 2 μM teniposide treatment for 48 hours enhanced the release and exposure of calreticulin.

[0061] FIG5 shows the IC50 values ​​of etoposide combined with IN10018 in colon cancer cells CT26.

[0062] FIG6 shows white light microscope pictures of colon cancer CT26 cells taken after incubation with drugs for 48 hours.

[0063] Figure 7 shows the situation after colon cancer CT26 cells were incubated with IN10018 and etoposide for 48 hours, wherein: 7a shows the percentage of CRT-positive cells after colon cancer CT26 cells were incubated with drugs for 48 hours; 7b shows the percentage of Annexin V-positive cells after colon cancer CT26 cells were incubated with drugs for 48 hours.

[0064] Figure 8 shows the situation after colon cancer CT26 cells were incubated with IN10018 and irinotecan for 48 hours, wherein: 8a shows the percentage of CRT-positive cells after colon cancer CT26 cells were incubated with drugs for 48 hours; 8b shows the percentage of Annexin V-positive cells after colon cancer CT26 cells were incubated with drugs for 48 hours.

[0065] FIG9 shows a curve showing the inhibitory effect of teniposide and its combination with IN10018 on the proliferation of colon cancer CT26 cells relative to drug concentration.

[0066] Figure 10 shows the situation after colon cancer CT26 cells were incubated with IN10018 and teniposide for 48 hours, wherein: 10a shows the percentage of Calreticulin (CRT)-positive cells in mouse colon cancer cell CT26 cells after incubation with drugs for 48 hours; 10b shows the percentage of Annexin-V-positive cells in mouse colon cancer cell CT26 cells after incubation with drugs for 48 hours.

[0067] FIG11 shows the tumor growth curve of the MC38 mouse colorectal cancer cell subcutaneous transplant tumor model in C57BL / 6 mice after administration of the drug, wherein the data points represent the mean tumor volume within the group, and the error bars represent the standard error (SEM).

[0068] FIG12 shows the body weight change curve of the MC38 mouse colorectal cancer cell C57BL / 6 mouse subcutaneous transplant tumor model after administration, wherein the data points represent the mean body weight within the group, and the error bars represent the standard error (SEM).

[0069] Figure 13 shows the situation after mouse breast cancer T41 cells were incubated with AMP945 and Etoposide for 48 hours, wherein: 13a shows the percentage of Calreticulin (CRT)-positive cells after mouse breast cancer T41 cells were incubated with drugs for 48 hours; 13b shows the percentage of Annexin-V-positive cells after mouse breast cancer T41 cells were incubated with drugs for 48 hours.

[0070] Figure 14 shows the situation after mouse breast cancer T41 cells were incubated with AMP945 and Irinotecan for 48 hours, wherein: 14a shows the percentage of Calreticulin (CRT)-positive cells after mouse breast cancer T41 cells were incubated with drugs for 48 hours; 14b shows the percentage of Annexin-V-positive cells after mouse breast cancer T41 cells were incubated with drugs for 48 hours. DETAILED DESCRIPTION

[0071] To make the purpose, technical solutions, and advantages of the embodiments of the present disclosure more clear, the technical solutions of the embodiments of the present disclosure will be clearly and completely described below in conjunction with the drawings of the embodiments of the present disclosure. Obviously, the described embodiments are part of the embodiments of the present disclosure, not all of the embodiments. Based on the described embodiments of the present disclosure, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0072] The present invention may be implemented in other specific forms without departing from the essential attributes of the present invention. It should be understood that, without conflict, any and all embodiments of the present invention may be combined with the technical features of any other embodiment or multiple other embodiments to produce additional embodiments. The present invention includes additional embodiments resulting from such combinations.

[0073] All publications and patents mentioned in this disclosure are hereby incorporated into the present disclosure in their entirety by reference. If the purposes or terms used in any publications and patents incorporated by reference conflict with the purposes or terms used in this disclosure, then the purposes and terms of this disclosure shall prevail.

[0074] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0075] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as those commonly used in the art to which the claimed subject matter belongs. If there are multiple definitions for a term, the definition herein shall prevail.

[0076] Except in the working examples or otherwise indicated, all numbers stating quantitative properties such as dosage in the specification and claims should be understood to be modified by the term "about" in all cases. It should also be understood that any numerical range recited herein is intended to include all subranges within the range and any combination of the various endpoints of the range or subrange.

[0077] As used in this disclosure, words such as "include," "comprising," or "including" mean that the elements preceding the word include the elements listed after the word and their equivalents, without excluding unlisted elements. The terms "comprising" or "including" as used herein may be open, semi-closed, or closed. In other words, the terms also include "consisting essentially of" or "consisting of."

[0078] definition

[0079] The following terms and symbols used in this application have the meanings described below, unless otherwise indicated in the context in which they are used. The term "FAK inhibitor" as used herein refers to an effective inhibitor of FAK, which can be suitable for mammals, particularly humans. In some embodiments, the FAK inhibitor is IN10018, Defactinib, GSK2256098, PF-00562271, VS-4718, AMP945, AMP886, or a pharmaceutically acceptable salt thereof, and the structure of IN10018 is as follows:

[0080] Defactinib, also known as defatinib, 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 Pharmaceuticals; and AMP945 has a CAS number of 1393653-34-3. In some embodiments, the FAK inhibitor is preferably IN10018, Defactinib, AMP945, or a pharmaceutically acceptable salt thereof. In some preferred embodiments, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, particularly IN10018 tartrate.

[0081] The term "topoisomerase inhibitor" as used herein refers to a class of compounds that can inhibit the activity of DNA topoisomerase, including topoisomerase I inhibitors and topoisomerase II inhibitors.

[0082] As used herein, the term "anthracycline" refers to a class of topoisomerase II inhibitors, including doxorubicin, epirubicin, daunorubicin, and aclarubicin.

[0083] The term "immune checkpoint inhibitor" as used herein refers to a drug that can increase the activity of the immune system by regulating the immune checkpoint pathway (e.g., PD-1, TIGIT, CTLA-4, LAG-3, TIM-3, etc.). In some embodiments, the immune checkpoint inhibitor is an antagonist of the PD-1 / PD-L1 (programmed cell death protein 1) pathway (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 this disclosure. For example, in the methods of treatment, drugs, and uses disclosed herein, the PD-1 / PD-L1 inhibitor is a PD-1 / PD-L1 antibody, including but not limited to pembrolizumab (Keytruda / K drug), tislelizumab (Tislelizumab / Baizean), nivolumab, toripalimab (Toripalimab / Tuoyi), atezolizumab (Atezolizumab / Tai Shengqi), durvalumab (Improvin / Durvalumab / Durvalumab), avelumab (Bavencio), atezolizumab (MPDL3280A / Atezolizumab / Tecentriq / T drug), BMS-936559 (fully human anti-PD-L1 IgG4 monoclonal antibody), GS-4224, AN-4005, or MX-10181. In some preferred embodiments, the PD-1 inhibitor is toripalimab. In some embodiments, PD-1 inhibitors are used to treat human subjects. In some embodiments, PD-1 is human PD-1. PD-1 / PD-L1 inhibitors also include small molecule PD-1 / PD-L1 inhibitors, such as INCB-086550, lazertinib, IMH-010, CA-170, ABSK043, or RRx-001. TIGIT (also known as WUCAM, Vstm3, and VSIG9) is a receptor of the Ig superfamily and a new immune checkpoint following PD-1 / PD-L1.For example, in the methods of treatment, drugs and uses disclosed herein, the immune checkpoint inhibitor is a TIGIT inhibitor, including but not limited to Ociperlimab / BGB-A1217, Vibostolimab, domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321. In some embodiments, the TIGIT inhibitor is used to treat human subjects. To avoid ambiguity, antibodies herein include bispecific antibodies.

[0084] As used herein, "drug combination" or "drug combination product" may refer to a fixed combination in the form of one dosage unit (for example, all active pharmaceutical ingredients are present in one dosage form) or a kit of parts for combined administration, or it may refer to a combination of one drug and instructions indicating that the drug can be used in combination with one or more other drugs.

[0085] As used herein, "combination therapy" or "combination drug" refers to the use of a drug in combination with one or more other drugs to treat a disease, including both the combination of a drug with one or more other drugs and the combination of a drug with instructions indicating that the drug can be used in combination with one or more other drugs.

[0086] "Administered simultaneously or sequentially" in this application refers to the administration of two or more drugs simultaneously or successively at certain time intervals within a dosing cycle (e.g., within 4 weeks, within 3 weeks, within 2 weeks, within 1 week, or within 24 hours), the mode of drug administration (e.g., oral, intravenous, intramuscular or subcutaneous administration, etc.) may be the same or different, and the administration frequency / cycle of two or more drugs may be the same or different. When the method of treatment, product, or use disclosed herein involves two drugs, the two drugs may be administered separately at the same time or at certain time intervals. When the method of treatment, product, or use disclosed herein involves three drugs, the three drugs may be administered at the same time point, or two drugs may be administered at one time point and the remaining drug may be administered at another time point, or all three drugs may be administered at different time points.

[0087] In some embodiments, the PD-1 / PD-L1 inhibitor is administered intravenously (e.g., as an intravenous infusion) or subcutaneously, or orally. Preferably, the PD-1 / PD-L1 inhibitor is administered as an intravenous infusion.

[0088] In some embodiments, the TIGIT inhibitor is administered intravenously (eg, as an intravenous infusion) or subcutaneously, or orally. Preferably, the TIGIT inhibitor is administered as an intravenous infusion.

[0089] The ability of immune checkpoint inhibitors to treat cancer depends on the presence of tumor antigen-specific T cells within tumor tissue. This requires that tumor tissue express antigens that distinguish itself from its non-transformed counterparts, for example, through novel protein products called neoantigens. Tumor neoantigen load is closely related to immunogenicity and sensitivity (for example, sensitivity to checkpoint inhibitor therapy), which means that tumors with poor immunogenicity should be largely resistant to these drugs. Therapies for releasing tumor antigens that can be taken up by APCs, such as those that induce immunogenic cell death (ICD), may promote effective anti-tumor immunity, especially when further combined with checkpoint inhibitors.

[0090] As used herein, the term "treat" refers to administering one or more pharmaceutical substances to a subject suffering from a disease or symptoms of a disease in order to cure, alleviate, relieve, alter, cure, ameliorate, improve, or affect the disease or symptoms of the disease. In some embodiments, the disease is a tumor or cancer.

[0091] The term "tumor" as used herein refers to an abnormal lesion formed when the cells of local tissues lose normal regulation of their growth at the genetic level under the influence of various tumorigenic factors, thereby causing abnormal proliferation of their clonal types. The tumors include, but are not limited to: bladder cancer, breast cancer, cervical cancer, colon cancer (including colorectal cancer), esophageal cancer, esophageal squamous cell carcinoma, head and neck cancer, liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer), 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, bile duct cancer, smooth muscle In some embodiments, the tumor is a breast cancer or colon cancer (including colorectal cancer).

[0092] As used herein, the term "subject" or "subject" refers to both mammals and non-mammals. Mammals refer to any member of the class mammalia, including but not limited to humans; non-human primates such as chimpanzees and other apes and monkey species; farm animals such as cattle, horses, sheep, goats, and pigs; livestock such as rabbits, dogs, and cats; laboratory animals, including rodents such as rats, mice, and guinea pigs; and the like. Examples of non-mammals include, but are not limited to, birds. The term "subject" does not limit the subject to a particular age or sex. In some embodiments, the subject is a human.

[0093] As used herein, the term "pharmaceutically acceptable" means non-toxic, biologically tolerable, and suitable for administration to a subject.

[0094] As used herein, the term "pharmaceutically acceptable salt" refers to non-toxic, biologically tolerable acid addition salts suitable for administration to a subject, including, but not limited to, 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 formate, acetate, malate, maleate, fumarate, tartrate, succinate, citrate, lactate, methanesulfonate, p-toluenesulfonate, 2-hydroxyethanesulfonate, benzoate, salicylate, stearate, and salts with the formula HOOC-(CH2) n-COOH (wherein n is 0-4) and the like.

[0095] In addition, 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 various synthetic methods that can be used to prepare non-toxic pharmaceutically acceptable acid addition salts without undue experimentation.

[0096] The term "pharmaceutically acceptable composition" as used herein means that it must be chemically and / or toxicologically compatible with the other ingredients comprising the formulation, and / or compatible with the subject being treated therewith. The term "therapeutically effective amount" as used herein means an amount that is generally sufficient to produce a beneficial therapeutic effect on the subject. The therapeutically effective amount of the present invention can be determined by conventional methods (e.g., modeling, dose escalation studies, or clinical trials) in combination with conventional influencing factors (e.g., route 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 drug, etc.).

[0097] As used herein, the term "inhibit" refers to a decrease in the baseline activity of a biological activity or process.

[0098] The term "kit" as used herein refers to a box for containing chemical reagents such as those for detecting chemical components, drug residues, and virus species. The kit of the present invention may include one, two, or three of (i) a FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor; and (ii) instructions, the instructions indicating that FAK inhibitors, topoisomerase inhibitors, and immune checkpoint inhibitors can be used to treat tumors in a subject. In one embodiment, the kit includes (i) a FAK inhibitor; and (ii) instructions, the instructions indicating that FAK inhibitors, topoisomerase inhibitors, and immune checkpoint inhibitors can be used to treat tumors in a subject. In one embodiment, the kit includes (i) a topoisomerase inhibitor; and (ii) instructions, the instructions indicating that FAK inhibitors, topoisomerase inhibitors, and immune checkpoint inhibitors can be used to treat tumors in a subject. In one embodiment, the kit includes (i) an immune checkpoint inhibitor; and (ii) instructions, the instructions indicating that FAK inhibitors, topoisomerase inhibitors, and immune checkpoint inhibitors can be used to treat tumors in a subject. In one embodiment, the kit includes (i) a FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor; and (ii) instructions indicating that a FAK inhibitor, a topoisomerase inhibitor, and an 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 a FAK inhibitor and a topoisomerase inhibitor can be used to treat a tumor in a subject. In one embodiment, the kit includes (i) a topoisomerase inhibitor; and (ii) instructions indicating that a topoisomerase inhibitor and a FAK inhibitor can be used to treat a tumor in a subject.

[0099] The compound of test kit can be contained in a separate container.Alternatively, two or more compounds are contained in the same container.For example, test kit can include a first container, a second container, a third container and a package insert, wherein the first container includes at least one dose of the medicine including a FAK inhibitor, the second container includes at least one dose of topoisomerase inhibitors, the third container includes at least one dose of the immune checkpoint inhibitor medicine, and the package insert includes instructions for using the tumor of the drug treatment object. The first container, the second container and the third container can include the same or different shapes (for example, vials, syringes and bottles) and / or materials (for example, plastic or glass). Test kit can also include other materials that can contribute to the administration of the drug, such as diluents, filters, IV bags and pipelines, needles and syringes.

[0100] The precise amount of a FAK inhibitor, topoisomerase inhibitor, or immune checkpoint inhibitor administered to a subject will depend on various factors, such as the given drug or compound, the pharmaceutical formulation, the route of administration, the type of disease, the condition, the identity of the subject or host being treated, etc., but can still be routinely determined by one skilled in the art. For example, determining an effective amount also depends on the extent, severity, and type of cell proliferation. A skilled person will be able to determine an appropriate dosage based on these and other factors.

[0101] FAK inhibitors, topoisomerase inhibitors, or immune checkpoint inhibitors can be administered by appropriate routes such as oral, intravenous, intramuscular, or subcutaneous administration.

[0102] For example, when administered orally, the drug can be administered orally with a pharmaceutically acceptable carrier, such as an inert diluent or an assimilable edible carrier. They can be encapsulated in hard-shell or soft-shell gelatin capsules, compressed into tablets, or mixed directly with the patient's food. For example, the drug can be combined with one or more excipients and used in the form of an ingestible tablet, buccal tablet, lozenge, capsule, elixir, suspension, syrup, or wafer. Tablets, lozenges, pills, capsules, etc. may further include: a binder, such as gum tragacanth, 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.

[0103] For example, when administered intravenously or intraperitoneally by infusion or injection, solutions of the drug can be prepared in water, optionally mixed with a nontoxic surfactant.

[0104] Exemplary pharmaceutical dosage forms for injection or infusion include sterile aqueous solutions, dispersions, or sterile powders containing the active ingredient, which are suitable for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions. In any case, the final dosage form should be sterile, fluid, and stable under the conditions of manufacture and storage.

[0105] Sterile injectable solutions can be prepared by incorporating the required amount of the drug into an appropriate solvent with the various other ingredients listed above as required, followed by filtered sterilization. For sterile powders for the preparation of sterile injectable solutions, the preferred preparation methods may be vacuum drying and freeze drying techniques, which can produce a powder of the active ingredient plus any other desired ingredients that have been previously sterile filtered.

[0106] The amount of FAK inhibitor, topoisomerase inhibitor, or immune checkpoint inhibitor required for treatment may vary not only with the specific agent selected, but also with the route of administration, the nature of the disease being treated, and the age and condition of the patient, and may ultimately be determined at the discretion of the attending physician or clinician. However, in general, the dosage may be in the range of about 0.1 to about 50 mg / kg body weight per day.

[0107] The FAK inhibitor is administered in a dosage range of 5 mg / day to 300 mg / day in adults. In a specific embodiment, IN10018 or a pharmaceutically acceptable salt thereof is administered in a dosage range of 5 mg / day to 100 mg / day in adults, for example, IN10018 or a pharmaceutically acceptable salt thereof is administered in a dosage range of 25 mg / day to 100 mg / day in adults, calculated as IN10018.

[0108] The topoisomerase inhibitor is administered at a dose of 1-300 mg / m2 per week in adults. 2 In a specific embodiment, irinotecan or a pharmaceutically acceptable salt thereof is administered at a dose range of 1-300 mg / m2 per week in adults. 2 The dosage is based on irinotecan; etoposide or its pharmaceutically acceptable salt is 1-300 mg / m per week in adults. 2 The dosage is based on etoposide; teniposide is 1-300 mg / m per week in adults. 2 The dosage is calculated based on teniposide.

[0109] The immune checkpoint inhibitor is administered at a dose of 2-10 mg / kg or 50-1200 mg in adults per administration, administered once every 2 to 3 weeks. In a specific embodiment, the immune checkpoint inhibitor is administered at a dose of 3-10 mg / kg or 100-1200 mg in adults per administration, administered once every 2 to 3 weeks.

[0110] Technical and scientific terms used herein without specific definition have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0111] In some embodiments, the present disclosure further discloses the following:

[0112] 1. A FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor for use in a method of treating a tumor in a subject, wherein the topoisomerase inhibitor is not an anthracycline.

[0113] 2. The FAK inhibitor, topoisomerase inhibitor, and immune checkpoint inhibitor according to embodiment 1, wherein the FAK inhibitor is IN10018, Defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886, or a pharmaceutically acceptable salt thereof, preferably IN10018, Defactinib, AMP945, or a pharmaceutically acceptable salt thereof, further preferably IN10018 or a pharmaceutically acceptable salt thereof, especially IN10018 tartrate, and the structure of IN10018 is as follows:

[0114] 3. The FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor according to embodiments 1-2, wherein the topoisomerase inhibitor is a topoisomerase I inhibitor.

[0115] 4. The FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor according to embodiment 3, wherein the topoisomerase I inhibitor is topotecan, irinotecan, belotecan, cositecan, exatecan (DX-8951), indenoisoquinoline, indotecan (LMP-400), indimitecan (LMP-776), simmitecan, gimatecan (ST1481), EC-112002, PLX-038 (NK012), AR-67, or a pharmaceutically acceptable salt thereof.

[0116] 5. The FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor according to any one of embodiments 3-4, wherein the topoisomerase I inhibitor is irinotecan, or a pharmaceutically acceptable salt thereof.

[0117] 6. The FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor according to any one of embodiments 1-2, wherein the topoisomerase inhibitor is a topoisomerase II inhibitor.

[0118] 7. The FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor as described in embodiment 6, wherein the topoisomerase II inhibitor is etoposide, teniposide, sabarubicin, or a pharmaceutically acceptable salt thereof.

[0119] 8. The FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor according to any one of embodiment 7, wherein the topoisomerase II inhibitor is etoposide, teniposide, or a pharmaceutically acceptable salt thereof.

[0120] 9. The FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor according to any one of embodiments 1-8, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, a PD-1 / PD-L1 small molecule inhibitor or a TIGIT antibody.

[0121] 10. The FAK inhibitor, topoisomerase inhibitor, and immune checkpoint inhibitor according to any one of embodiments 1 to 9, wherein 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, or ab), durvalumab, avelumab, atezolizumab, camrelizumab, sintilimab, cemiplimab, envafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005, or MX-10181.

[0122] 11. The use, pharmaceutical combination product, or method according to any one of embodiments 1 to 9, wherein the immune checkpoint inhibitor is a PD-1 / PD-L1 small molecule inhibitor, and further, the PD-1 / PD-L1 small molecule inhibitor is INCB-086550, lazertinib, IMH-010, CA-170, ABSK043, or RRx-001.

[0123] 12. The FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor according to any one of embodiments 1 to 9, wherein the immune checkpoint inhibitor is a TIGIT antibody, and further, the TIGIT antibody is Ociperlimab (Ociperlimab / BGB-A1217), Vibostolimab, domvanalimab (AB154), Tiragolumab, Belre stotug, 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.

[0124] 13. The FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor according to embodiment 1, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; the topoisomerase inhibitor is irinotecan 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, especially an anti-PD-1 / PD-L1 antibody.

[0125] 14. The FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor as described in Embodiment 1, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; the topoisomerase inhibitor is etoposide, teniposide, 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, especially an anti-PD-1 / PD-L1 antibody.

[0126] 15. The FAK inhibitor, topoisomerase inhibitor, and immune checkpoint inhibitor of any one of embodiments 1-14, wherein the FAK inhibitor, the topoisomerase inhibitor, and the immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.

[0127] 16. A FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor as described in any one of embodiments 1-15, wherein 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, bile duct cancer, leiomyosarcoma, liposarcoma, nasopharyngeal carcinoma , neuroendocrine cancer, ovarian cancer, salivary gland cancer, metastases caused by spindle cell cancer, anaplastic large cell lymphoma, undifferentiated thyroid cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma, glioma or hematological malignancies, such as acute myeloid leukemia (AML), acute lymphocytic leukemia (ALL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML); preferably, 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.

[0128] 17. A FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor as described in any one of embodiments 1-16, wherein the tumor is breast cancer or colon cancer (including colorectal cancer).

[0129] 18. A kit or pharmaceutically acceptable composition comprising:

[0130] (a) FAK inhibitors;

[0131] (b) a topoisomerase inhibitor, said topoisomerase inhibitor not being an anthracycline; and

[0132] (c) Immune checkpoint inhibitors.

[0133] 19. The kit or composition according to embodiment 18, wherein the FAK inhibitor is IN10018, Defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886, or a pharmaceutically acceptable salt thereof, preferably IN10018, Defactinib, AMP945, or a pharmaceutically acceptable salt thereof, further preferably IN10018 or a pharmaceutically acceptable salt thereof, especially IN10018 tartrate, wherein the structure of IN10018 is as follows:

[0134] 20. The kit or composition of embodiments 18-19, wherein the topoisomerase inhibitor is a topoisomerase I inhibitor.

[0135] 21. The kit or composition of embodiment 20, wherein the topoisomerase I inhibitor is topotecan, irinotecan, belotecan, cositecan, exatecan (DX-8951), indenoisoquinoline, indotecan (LMP-400), indimitecan (LMP-776), simmitecan, gimatecan (ST1481), EC-112002, PLX-038 (NK012), AR-67, or a pharmaceutically acceptable salt thereof.

[0136] 22. The kit or composition of any one of embodiments 20-21, wherein the topoisomerase I inhibitor is irinotecan, or a pharmaceutically acceptable salt thereof.

[0137] 23. The kit or composition of any one of embodiments 18-19, wherein the topoisomerase inhibitor is a topoisomerase II inhibitor.

[0138] 24. The kit or composition of embodiment 23, wherein the topoisomerase II inhibitor is etoposide, teniposide, sabarubicin, or a pharmaceutically acceptable salt thereof.

[0139] 25. The kit or composition of embodiment 24, wherein the topoisomerase II inhibitor is etoposide, teniposide, or a pharmaceutically acceptable salt thereof.

[0140] 26. The kit or composition of any one of embodiments 18-25, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, a PD-1 / PD-L1 small molecule inhibitor, or a TIGIT antibody.

[0141] 27. The kit or composition of any one of embodiments 18-26, wherein 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, Atezolizumab, Camrelizumab, Sintilimab, Cemiplimab, envafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005, or MX-10181.

[0142] 28. The use, pharmaceutical combination product, or method according to any one of embodiments 18 to 26, wherein the immune checkpoint inhibitor is a PD-1 / PD-L1 small molecule inhibitor, and further, the PD-1 / PD-L1 small molecule inhibitor is INCB-086550, lazertinib, IMH-010, CA-170, ABSK043, or RRx-001.

[0143] 29. The kit or composition of any one of embodiments 18-26, wherein the immune checkpoint inhibitor is a TIGIT antibody, and further, the TIGIT antibody is Ociperlimab (BGB-A1217), Vibostolimab, domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036, or IBI-321.

[0144] 30. The kit or composition according to embodiment 18, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; the topoisomerase inhibitor is irinotecan 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, especially an anti-PD-1 / PD-L1 antibody.

[0145] 31. The kit or composition according to embodiment 18, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; the topoisomerase inhibitor is etoposide, teniposide, 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, especially an anti-PD-1 / PD-L1 antibody.

[0146] 32. The kit or composition of any one of embodiments 18-31, wherein the composition is for use in medicine.

[0147] 33. The kit or composition of embodiment 32, wherein the drug is used to treat a tumor, and 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, bile duct cancer, leiomyosarcoma, liposarcoma, nasopharyngeal cancer, neuroendocrine cancer, The invention relates to a method for treating a thyroid cancer of the present invention and the like. The method comprises the following steps: treating a thyroid cancer of the present invention and the like; wherein the thyroid cancer is a thyroid cancer, a thyroid cancer, a salivary gland cancer, a metastatic tumor caused by spindle cell carcinoma, an anaplastic large cell lymphoma, an undifferentiated thyroid cancer, a non-Hodgkin's lymphoma, a Hodgkin's lymphoma, a 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), chronic myeloid leukemia (CML); preferably, 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.

[0148] 34. The kit or composition of embodiment 33, wherein the tumor is breast cancer or colon cancer (including colorectal cancer).

[0149] 35. A method of treating a tumor in a subject, wherein the method comprises administering to the subject a therapeutically effective amount of a FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

[0150] 36. The method according to embodiment 35, wherein the FAK inhibitor is IN10018, Defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886, or a pharmaceutically acceptable salt thereof, preferably IN10018, Defactinib, AMP945, or a pharmaceutically acceptable salt thereof, further preferably IN10018 or a pharmaceutically acceptable salt thereof, especially IN10018 tartrate, wherein the structure of IN10018 is as follows:

[0151] 37. The method of embodiments 35-36, wherein the topoisomerase inhibitor is a topoisomerase I inhibitor.

[0152] 38. The method of embodiment 37, wherein the topoisomerase I inhibitor is topotecan, irinotecan, belotecan, cositecan, exatecan (DX-8951), indenoisoquinoline, indotecan (LMP-400), indimitecan (LMP-776), simmitecan, gimatecan (ST1481), EC-112002, PLX-038 (NK012), AR-67, or a pharmaceutically acceptable salt thereof.

[0153] 39. The method according to embodiment 38, wherein the topoisomerase I inhibitor is irinotecan, or a pharmaceutically acceptable salt thereof.

[0154] 40. The method of any one of embodiments 35-36, wherein the topoisomerase inhibitor is a topoisomerase II inhibitor.

[0155] 41. The method according to embodiment 40, wherein the topoisomerase II inhibitor is etoposide, teniposide, sabarubicin, or a pharmaceutically acceptable salt thereof.

[0156] 42. The method according to embodiment 41, wherein the topoisomerase II inhibitor is etoposide, teniposide, or a pharmaceutically acceptable salt thereof.

[0157] 43. A method as described in any one of embodiments 35-42, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, a PD-1 / PD-L1 small molecule inhibitor, or a TIGIT antibody.

[0158] 44. The method according to any one of embodiments 35 to 43, wherein 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, Atezolizumab, Camrelizumab, Sintilimab, Cemiplimab, envafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.

[0159] 45. The use, pharmaceutical combination product, or method according to any one of embodiments 35 to 43, wherein the immune checkpoint inhibitor is a PD-1 / PD-L1 small molecule inhibitor, and further, the PD-1 / PD-L1 small molecule inhibitor is INCB-086550, lazertinib, IMH-010, CA-170, ABSK043, or RRx-001.

[0160] 46. ​​The method according to any one of embodiments 35 to 43, wherein the immune checkpoint inhibitor is a TIGIT antibody, and further, the TIGIT antibody is Ociperlimab (BGB-A1217), Vibostolimab, domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.

[0161] 47. The method according to embodiment 35, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; the topoisomerase inhibitor is irinotecan 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, especially an anti-PD-1 / PD-L1 antibody.

[0162] 48. The method according to embodiment 35, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; the topoisomerase inhibitor is etoposide, teniposide, 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, especially an anti-PD-1 / PD-L1 antibody.

[0163] 49. The method of any one of embodiments 35-48, wherein the FAK inhibitor, the topoisomerase inhibitor, and the immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.

[0164] 50. A method as described in any one of embodiments 35-49, wherein 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, bile duct cancer, leiomyosarcoma, liposarcoma, nasopharyngeal cancer, neuroendocrine cancer, ovarian cancer Preferably, the tumor is a breast cancer, an ovarian cancer, a colon cancer (including colorectal cancer), a lung cancer (including small cell lung cancer and non-small cell lung cancer), a melanoma or a pancreatic cancer.

[0165] 51. The method of any one of embodiments 35-50, wherein the tumor is breast cancer or colon cancer (including colorectal cancer).

[0166] 52. A FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor for use in a method of treating a tumor by increasing immunogenic cell death in a subject, wherein the topoisomerase inhibitor is not an anthracycline.

[0167] 53. The FAK inhibitor, topoisomerase inhibitor, and immune checkpoint inhibitor according to embodiment 52, wherein the FAK inhibitor is IN10018, Defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886, or a pharmaceutically acceptable salt thereof, preferably IN10018, Defactinib, AMP945, or a pharmaceutically acceptable salt thereof, further preferably IN10018 or a pharmaceutically acceptable salt thereof, especially IN10018 tartrate, and the structure of IN10018 is as follows:

[0168] 54. The FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor of embodiments 52-53, wherein the topoisomerase inhibitor is a topoisomerase I inhibitor.

[0169] 55. A FAK inhibitor, a topoisomerase inhibitor and an immune checkpoint inhibitor as described in embodiment 54, wherein the topoisomerase I inhibitor is topotecan, irinotecan, Belotecan, Cositecan, Exatecan (DX-8951), Indenoisoquinoline, Indotecan (LMP-400), Indimitecan (LMP-776), Simmitecan, Gimatecan (ST1481), EC-112002, PLX-038 (NK012), AR-67, or a pharmaceutically acceptable salt thereof.

[0170] 56. The FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor according to embodiment 55, wherein the topoisomerase I inhibitor is irinotecan or a pharmaceutically acceptable salt thereof.

[0171] 57. A FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor as described in any one of embodiments 52-53, wherein the topoisomerase inhibitor is a topoisomerase II inhibitor.

[0172] 58. The FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor according to embodiment 57, wherein the topoisomerase II inhibitor is etoposide, teniposide, sabarubicin, or a pharmaceutically acceptable salt thereof.

[0173] 59. The FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor according to any one of embodiment 58, wherein the topoisomerase II inhibitor is etoposide, teniposide, or a pharmaceutically acceptable salt thereof.

[0174] 60. A FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor as described in any one of embodiments 52-59, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, a PD-1 / PD-L1 small molecule inhibitor, or a TIGIT antibody.

[0175] 61. The FAK inhibitor, topoisomerase inhibitor, and immune checkpoint inhibitor according to any one of embodiments 52-60, wherein 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, or the like. mab), durvalumab, avelumab, atezolizumab, camrelizumab, sintilimab, cemiplimab, envafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005, or MX-10181.

[0176] 62. The use, pharmaceutical combination product, or method according to any one of embodiments 52 to 60, wherein the immune checkpoint inhibitor is a PD-1 / PD-L1 small molecule inhibitor, and further, the PD-1 / PD-L1 small molecule inhibitor is INCB-086550, Lazertinib, IMH-010, CA-170, ABSK043, or RRx-001.

[0177] 63. The FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor according to any one of embodiments 52-60, wherein the immune checkpoint inhibitor is a TIGIT antibody, and further, the TIGIT antibody is Ociperlimab (Ociperlimab / BGB-A1217), Vibostolimab, domvanalimab (AB154), Tiragolumab, Belr estotug, 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.

[0178] 64. The FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor according to embodiment 52, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; the topoisomerase inhibitor is irinotecan 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, especially an anti-PD-1 / PD-L1 antibody.

[0179] 65. The FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor according to embodiment 52, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; the topoisomerase inhibitor is etoposide, teniposide, 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, especially an anti-PD-1 / PD-L1 antibody.

[0180] 66. The FAK inhibitor, topoisomerase inhibitor, and immune checkpoint inhibitor of any one of embodiments 52-65, wherein the FAK inhibitor, the topoisomerase inhibitor, and the immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.

[0181] 67. A FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor as described in any one of embodiments 52-66, wherein 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, bile duct cancer, leiomyosarcoma, liposarcoma, nasopharyngeal cancer Preferably, 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.

[0182] 68. A FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor as described in any one of embodiments 52-67, wherein the tumor is breast cancer or colon cancer (including colorectal cancer).

[0183] 69. A method of treating a tumor by increasing immunogenic cell death in a subject, wherein the method comprises administering to the subject a therapeutically effective amount of a FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

[0184] 70. The method according to embodiment 69, wherein the FAK inhibitor is IN10018, Defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886, or a pharmaceutically acceptable salt thereof, preferably IN10018, Defactinib, AMP945, or a pharmaceutically acceptable salt thereof, further preferably IN10018 or a pharmaceutically acceptable salt thereof, especially IN10018 tartrate, and the structure of IN10018 is as follows:

[0185] 71. The method of embodiments 69-70, wherein the topoisomerase inhibitor is a topoisomerase I inhibitor.

[0186] 72. A method as described in any one of embodiments 69-71, wherein the topoisomerase I inhibitor is topotecan, irinotecan, belotecan, cositecan, exatecan (DX-8951), indenoisoquinoline, indotecan (LMP-400), indimitecan (LMP-776), simmitecan, gimatecan (ST1481), EC-112002, PLX-038 (NK012), AR-67, or a pharmaceutically acceptable salt thereof.

[0187] 73. The method of embodiment 72, wherein the topoisomerase I inhibitor is irinotecan, or a pharmaceutically acceptable salt thereof.

[0188] 74. The method of any one of embodiments 69-70, wherein the topoisomerase inhibitor is a topoisomerase II inhibitor.

[0189] 75. The method of embodiment 74, wherein the topoisomerase II inhibitor is etoposide, teniposide, sabarubicin, or a pharmaceutically acceptable salt thereof.

[0190] 76. The method according to embodiment 75, wherein the topoisomerase II inhibitor is etoposide, teniposide, or a pharmaceutically acceptable salt thereof.

[0191] 77. The method of any one of embodiments 69-76, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, a PD-1 / PD-L1 small molecule inhibitor, or a TIGIT antibody.

[0192] 78. The method of any one of embodiments 69-77, wherein 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, Atezolizumab, Camrelizumab, Sintilimab, Cemiplimab, envafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.

[0193] 79. The use, pharmaceutical combination product, or method according to any one of embodiments 69 to 77, wherein the immune checkpoint inhibitor is a PD-1 / PD-L1 small molecule inhibitor, and further, the PD-1 / PD-L1 small molecule inhibitor is INCB-086550, Lazertinib, IMH-010, CA-170, ABSK043, or RRx-001.

[0194] 80. The method according to any one of embodiments 69 to 77, wherein the immune checkpoint inhibitor is a TIGIT antibody, and further, the TIGIT antibody is Ociperlimab (BGB-A1217), Vibostolimab, domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.

[0195] 81. The method according to embodiment 69, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; the topoisomerase inhibitor is irinotecan 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, especially an anti-PD-1 / PD-L1 antibody.

[0196] 82. The method according to embodiment 69, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; the topoisomerase inhibitor is etoposide, teniposide, 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, especially an anti-PD-1 / PD-L1 antibody.

[0197] 83. The method of any one of embodiments 69-82, wherein the FAK inhibitor, the topoisomerase inhibitor, and the immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.

[0198] 84. A method as described in any one of embodiments 69-83, wherein 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, bile duct cancer, leiomyosarcoma, liposarcoma, nasopharyngeal cancer, neuroendocrine cancer, ovarian cancer Preferably, the tumor is a breast cancer, an ovarian cancer, a colon cancer (including colorectal cancer), a lung cancer (including small cell lung cancer and non-small cell lung cancer), a melanoma or a pancreatic cancer.

[0199] 85. The method of any one of embodiments 69-84, wherein the tumor is breast cancer or colon cancer (including colorectal cancer).

[0200] 86. Use of a FAK inhibitor in the preparation of a medicament for treating a tumor in a subject, wherein the FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor are administered to the subject, wherein the topoisomerase inhibitor is not an anthracycline.

[0201] 87. Use of a topoisomerase inhibitor in the preparation of a medicament for treating a tumor in a subject, wherein a FAK inhibitor, the topoisomerase inhibitor, and an immune checkpoint inhibitor are administered to the subject, wherein the topoisomerase inhibitor is not an anthracycline.

[0202] 88. Use of an immune checkpoint inhibitor in the preparation of a medicament for treating a tumor in a subject, wherein a FAK inhibitor, a topoisomerase inhibitor, and the immune checkpoint inhibitor are administered to the subject, wherein the topoisomerase inhibitor is not an anthracycline.

[0203] 89. Use of a FAK inhibitor, a topoisomerase inhibitor and an immune checkpoint inhibitor in the preparation of a combination drug for treating tumors, wherein the topoisomerase inhibitor is not an anthracycline.

[0204] 90. Use of a FAK inhibitor in the preparation of a combination drug for treating tumors with a topoisomerase inhibitor and an immune checkpoint inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

[0205] 91. Use of a topoisomerase inhibitor in the preparation of a combination drug for treating tumors with a FAK inhibitor and an immune checkpoint inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

[0206] 92. Use of an immune checkpoint inhibitor in the preparation of a combination drug for treating tumors with a FAK inhibitor and a topoisomerase inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

[0207] 93. Use of a FAK inhibitor, a topoisomerase inhibitor and an immune checkpoint inhibitor in the preparation of a medicament for combined treatment of tumors, wherein the topoisomerase inhibitor is not an anthracycline.

[0208] 94. Use of a FAK inhibitor in the preparation of a medicament for treating tumors in combination with a topoisomerase inhibitor and an immune checkpoint inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

[0209] 95. Use of a topoisomerase inhibitor in the preparation of a medicament for treating tumors in combination with a FAK inhibitor and an immune checkpoint inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

[0210] 96. Use of an immune checkpoint inhibitor in the preparation of a medicament for treating tumors in combination with a FAK inhibitor and a topoisomerase inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

[0211] 97. The use according to any one of embodiments 86-96, wherein the FAK inhibitor is IN10018, Defactinib, GSK2256098, PF-00562271, APG-2449, VS-4718, AMP945, AMP886, or a pharmaceutically acceptable salt thereof, preferably IN10018, Defactinib, AMP945, or a pharmaceutically acceptable salt thereof, further preferably IN10018 or a pharmaceutically acceptable salt thereof, especially IN10018 tartrate, and the structure of IN10018 is as follows:

[0212] 98. The use according to any one of embodiments 86-97, wherein the topoisomerase inhibitor is a topoisomerase I inhibitor.

[0213] 99. The use according to embodiment 98, wherein the topoisomerase I inhibitor is topotecan, irinotecan, belotecan, cositecan, exatecan (DX-8951), indenoisoquinoline, indotecan (LMP-400), indimitecan (LMP-776), simmitecan, gimatecan (ST1481), EC-112002, PLX-038 (NK012), AR-67, or a pharmaceutically acceptable salt thereof.

[0214] 100. The use according to embodiment 99, wherein the topoisomerase I inhibitor is irinotecan or a pharmaceutically acceptable salt thereof.

[0215] 101. The use according to any one of embodiments 86-97, wherein the topoisomerase inhibitor is a topoisomerase II inhibitor.

[0216] 102. The use according to embodiment 101, wherein the topoisomerase II inhibitor is etoposide, teniposide, sabarubicin, or a pharmaceutically acceptable salt thereof.

[0217] 103. The use according to embodiment 102, wherein the topoisomerase II inhibitor is etoposide, teniposide, or a pharmaceutically acceptable salt thereof.

[0218] 104. The use according to any one of embodiments 86-103, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, a PD-1 / PD-L1 small molecule inhibitor, or a TIGIT antibody.

[0219] 105. The use according to any one of embodiments 86 to 104, wherein 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, Atezolizumab, Camrelizumab, Sintilimab, Cemiplimab, envafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.

[0220] 106. The use, pharmaceutical combination product, or method according to any one of embodiments 86 to 104, wherein the immune checkpoint inhibitor is a PD-1 / PD-L1 small molecule inhibitor, and further, the PD-1 / PD-L1 small molecule inhibitor is INCB-086550, Lazertinib, IMH-010, CA-170, ABSK043, or RRx-001.

[0221] 107. The use according to any one of embodiments 86 to 104, wherein the immune checkpoint inhibitor is a TIGIT antibody, and further, the TIGIT antibody is Ociperlimab (BGB-A1217), Vibostolimab, domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.

[0222] 108. The use according to any one of embodiments 86-96, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; the topoisomerase inhibitor is irinotecan 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, especially an anti-PD-1 / PD-L1 antibody.

[0223] 109. The use according to any one of embodiments 86-96, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; the topoisomerase inhibitor is etoposide, teniposide, or a pharmaceutically acceptable salt thereof; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, especially an anti-PD-1 / PD-L1 antibody.

[0224] 110. The use according to any one of embodiments 86-109, wherein the FAK inhibitor, the topoisomerase inhibitor, and the immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.

[0225] 111. The use according to any one of embodiments 86 to 110, wherein 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, bile duct cancer, leiomyosarcoma, liposarcoma, nasopharyngeal cancer, neuroendocrine cancer, ovariectomy, or the like. Preferably, 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 breast cancer or colon cancer (including colorectal cancer).

[0227] Example

[0228] The following examples are provided to further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0229] The experimental methods in the following examples without specifying specific conditions can be carried out according to conventional conditions of such reactions or according to conditions recommended by the manufacturers.

[0230] Unless otherwise specified, the experimental materials and reagents used in the following examples can be obtained from commercial channels.

[0231] The abbreviations used in the examples have the following meanings:

[0232] Example 1: Study on the Enhancement of Cell Death of Breast Cancer Cells Against Etoposide Immunogenicity by FAK Target Inhibition

[0233] Experimental plan:

[0234] 1. FAK silencing combined with etoposide promotes apoptosis of breast cancer cells MDA-MB-231

[0235] In vitro experiments used siRNA technology to reduce the expression level of FAK and combined it with etoposide for 48 hours; flow cytometry was used to detect apoptosis in control and FAK-silenced cells.

[0236] 2. FAK silencing combined with etoposide can effectively promote ICD in breast cancer cells MDA-MB-231

[0237] In vitro experiments used siRNA technology to reduce the expression level of FAK and combined it with etoposide for 48 hours; flow cytometry was used to detect the expression of calreticulin, the main target of ICD.

[0238] Experimental antibody: recombinant Alexa 647 fluorescent Anti-Calreticulin antibody (Abcam, ab196159), Annexin V apoptosis detection kit (Invitrogen, A35110).

[0239] FAK siRNA was provided by Genema, and the sequence is shown in the following table:

[0240] F: CCUGUAUGCCUAUCAGCUUTT;

[0241] R:AAGCUGAUAGGCAUACAGGTT

[0242] Experimental instruments:

[0243] Fluorescence microscope (Olympus U-HGLGPS) and chemiluminescence imager (BIORAD chemidoc touch)

[0244] Experimental results:

[0245] 1. FAK silencing combined with etoposide significantly increased apoptosis in MDA-MB-231 cells

[0246] MDA-MB-231 cells were transfected with control siRNA or FAK siRNA at a final concentration of 50 nM. Twenty-four hours after transfection, cells were treated with 2 μM etoposide for 48 hours. Cells were stained using an Annexin V kit and analyzed by flow cytometry. Early and late apoptosis values ​​were compared with those in the DMSO control group and plotted using Graphpad 8.0. The results showed that etoposide treatment significantly enhanced apoptosis in the FAK silencing group compared to the control group, as shown in Figure 1.

[0247] 2. FAK silencing combined with etoposide significantly upregulated the release and exposure of the ICD target calreticulin

[0248] MDA-MB-231 cells were transfected with either control siRNA or FAK siRNA at a final concentration of 50 nM. Twenty-four hours after transfection, cells were treated with 2 μM etoposide for 48 hours. Calreticulin was then fluorescently stained with an antibody and analyzed by flow cytometry. FlowJo software analysis revealed that FAK silencing combined with etoposide significantly enhanced calreticulin release and exposure, as shown in Figure 2.

[0249] Example 2: Study on the Enhancement of Teniposide-Induced Cell Death in Breast Cancer Cells by FAK Target Inhibition

[0250] Experimental plan:

[0251] 1. FAK silencing combined with teniposide promotes apoptosis of breast cancer cells MDA-MB-231

[0252] In vitro experiments used siRNA technology to reduce the expression level of FAK and combined it with teniposide for 48 hours; flow cytometry was used to detect apoptosis in control and FAK-silenced cells.

[0253] 2. FAK silencing combined with teniposide can effectively promote ICD in breast cancer cells MDA-MB-231

[0254] In vitro experiments used siRNA technology to reduce the expression level of FAK and combined it with teniposide for 48 hours; flow cytometry was used to detect the expression of calreticulin, the main target of ICD.

[0255] Experimental Antibodies:

[0256] Restructuring Alexa 647 fluorescent Anti-Calreticulin antibody (Abcam, ab196159), Annexin V apoptosis detection kit (Invitrogen, A35110).

[0257] FAK siRNA was provided by Genema, and the sequence is shown in the following table:

[0258] F: CCUGUAUGCCUAUCAGCUUTT;

[0259] R:AAGCUGAUAGGCAUACAGGTT

[0260] Experimental instruments:

[0261] Fluorescence microscope (Olympus U-HGLGPS) and chemiluminescence imager (BIORAD chemidoc touch).

[0262] Experimental results:

[0263] 1. FAK silencing combined with teniposide significantly increased apoptosis in MDA-MB-231 cells

[0264] MDA-MB-231 cells were transfected with either control siRNA or FAK siRNA at a final concentration of 50 nM. Twenty-four hours after transfection, cells were treated with 2 μM teniposide for 48 hours. Cells were stained using an Annexin V kit and analyzed by flow cytometry. Early and late apoptosis were compared with those in the DMSO control group and plotted using Graphpad 8.0. The results showed that apoptosis was significantly enhanced in the FAK silencing group after teniposide treatment compared to the control group, as shown in Figure 3.

[0265] 2. FAK silencing combined with teniposide significantly upregulated the release and exposure of the ICD target calreticulin

[0266] MDA-MB-231 cells were transfected with control siRNA or FAK siRNA at a final concentration of 50 nM. Twenty-four hours after transfection, cells were treated with 2 μM teniposide for 48 hours. Calreticulin was then fluorescently stained with an antibody and analyzed by flow cytometry. FlowJo software analysis revealed that FAK silencing combined with teniposide significantly enhanced calreticulin release and exposure, as shown in Figure 4.

[0267] Example 3: Synergistic effect of IN10018 and etoposide in colon cancer CT26 cells

[0268] CT26 cells were cultured in RPMI 1640 (Shanghai Yuanpei, catalog number: L210KJ, batch number: F210916) + 10% FBS (Gibco, catalog number: 10099-141c, batch number: 2158737cp) and passaged twice. When cells were in good condition, they were plated in 96-well plates at 3,000 cells per well. After 24 hours of cell spreading, medium containing etoposide was added. Ten drug concentrations were set up, starting with a 30 μM concentration, diluted fivefold, and ending with a control with a drug concentration of 0. Three wells were set up for each drug concentration. Simultaneously, another set of cells was established using the same drug concentrations as above. However, 5 μM IN10018 was added to each well, the drugs were mixed, and the cells were cultured at 37°C in a 5% CO2 incubator for 72 hours.

[0269] After 72 hours of drug exposure, cells were observed under a microscope. 10 μl of CCK8 detection reagent (Cellorlab, catalog number: CX001M, batch number: 2571100) was added to each well and incubated in a 37°C, 5% CO2 incubator for 2-4 hours. The plate was then read using a microplate reader (chemiluminescence) at OD450. Analysis revealed that the IC50 for the etoposide (source: MCE, catalog number: 33419-42-0, batch number: 113741) group was 24.43 μM; the IC50 for the etoposide + 5 μM IN10018 group was 4.86 μM. The IC50 for the group containing IN10018 was significantly lower than that for the group without IN10018, indicating that the combined treatment group was more effective than the monotherapy group, as shown in Figure 5.

[0270] Example 4: Study of IN10018 and Etoposide in Colon Cancer CT26 Cells

[0271] CT26 cells (Institute of Cell Biology, Chinese Academy of Sciences) were cultured in RPMI 1640 (Shanghai Yuanpei, catalog number: L210KJ, batch number: F210916) + 10% FBS (Gibco, catalog number: 10099-141c, batch number: 2158737cp) and passaged twice. When cells were in good condition, the culture medium was plated in 24-well plates. After 24 hours of cell expansion, four groups were assigned: the first group served as the control group and received culture medium; the second group received IN10018 at a concentration of 5 μM; the third group received etoposide (from MCE, catalog number: 33419-42-0, batch number: 113741) at a concentration of 20 μM; and the fourth group received a combination of IN10018 (5 μM) and etoposide (20 μM). The drugs were mixed and incubated at 37°C in a 5% CO2 incubator for 48 hours.

[0272] After 48 hours of drug treatment, cells were observed under a microscope and photographed, and the photographs were saved. Cells were then collected for flow cytometry analysis and washed twice with flow cytometry buffer (PBS + 2% FBS). 0.5 μl of AF647 anti-calreticulin antibody (abcam, catalog number: ab196159, batch number: CR33676773) was added to each well, mixed thoroughly, and incubated at 4°C in the dark for 20 minutes. After 20 minutes, cells were washed twice with flow cytometry buffer (PBS + 2% FBS). Using the apoptosis detection kit (Beyotime, catalog number: CL062L, batch number: 021921210811), 195 μl of annexin V-FITC conjugate was added, the cells were gently mixed, and then 5 μl of annexin V-FITC antibody was added and gently mixed. 10 μl of PI dye was added, mixed gently, and incubated at room temperature in the dark for 15 minutes before analysis on a flow cytometer.

[0273] Microscopic observation of cells revealed poor cell status in the etoposide monotherapy and dual-drug groups, with the dual-drug group showing the worst results, with more cell death. Cells in the control and IN10018 groups remained in good condition. Results showed that the CRT and Annexin V positivity rates in the dual-drug group were higher than in the monotherapy group (see Figures 6 and 7 for details).

[0274] Example 5: Study on the induction of immunogenic cell death targets by irinotecan and IN10018 in mouse colon cancer CT26 cells in vitro

[0275] Grouping information is shown in Table 1.

[0276] Table 1: Grouping scheme for in vitro efficacy experiments

[0277] The information of the tested compounds is shown in Table 2.

[0278] Table 2: Information of test compounds

[0279] The main experimental reagents are shown in Table 3.

[0280] Table 3: Main reagents for the experiment

[0281] Experimental methods and steps

[0282] Cell culture

[0283] CT26 cells were maintained and passaged by Ins Biotech (Nanjing) Co., Ltd. Cells were cultured as monolayers in RPMI1640 medium supplemented with 10% fetal bovine serum in a 37°C, 5% CO2 incubator. Cells were routinely digested and passaged using trypsin two to three times a week. When cells reached 80%-90% confluence during the exponential growth phase, they were harvested and plated.

[0284] Cell plating

[0285] CT26 cells were trypsinized, harvested, and counted. Based on the count, the cells were diluted with the appropriate complete culture medium to a concentration of 50,000 cells / mL. The cells were then plated onto 24-well cell culture plates, with 1 mL of the cell suspension per well (i.e., 50,000 cells). After plating, the cells were cultured in a 37°C, 5% CO2 incubator.

[0286] Add the test compound

[0287] 24 hours after plating, the test compounds, irinotecan and IN10018, were added to different wells. The test compounds were prepared and aliquoted in DMSO at a volume of 50 μL per vial and stored at -20°C in the dark. For this experiment, one vial was removed for drug addition and testing. Groups and drug concentrations are shown in Table 4.

[0288] Table 4: Drug groups and dosing concentrations

[0289] Collect cells for flow cytometry

[0290] After washing the cells twice with flow cytometry buffer (DPBS + 2% FBS), 0.5 μL of AF647 Anti-Calreticulin Antibody was added to each well, mixed, and incubated at 4°C in the dark for 20 minutes. After washing once with flow cytometry buffer, 195 μL of Annexin-V-FITC conjugate was added. After gently mixing the cells, 5 μL of Annexin-V-FITC antibody was added. After further mixing, 10 μL of PI dye was added and mixed. The cells were incubated at room temperature in the dark for 15 minutes before flow cytometry analysis.

[0291] Data Analysis

[0292] After the experiment, Flowjo (V10) software was used to analyze the cell positive rate.

[0293] Experimental results

[0294] 48 hours after the drug treatment, flow cytometric analysis results showed that the positive rates of CRT and Annexin V in the combination drug group were significantly higher than those in the single drug group, as shown in Figure 8.

[0295] Example 6: Study on the Inhibitory Activity of Teniposide and IN10018 on the Proliferation of Mouse Colon Cancer CT26 Cells in Vitro

[0296] Experimental design

[0297] See Table 5 for grouping information.

[0298] Table 5: Grouping scheme for cell proliferation inhibition assay

[0299] The information of the tested compounds is shown in Table 6.

[0300] Table 6:

[0301] The main reagent information is shown in Table 7.

[0302] Table 7:

[0303] Experimental methods and steps:

[0304] Cell culture:

[0305] CT26 cells were revived and maintained by Inspiron Biotech (Nanjing) Co., Ltd. Cells were cultured as adherent monolayers in RPMI1640 medium supplemented with 10% fetal bovine serum at 37°C and 5% CO2. Cells were trypsinized and passaged two to three times weekly. When cells reached 80%-90% confluence during the exponential growth phase, they were harvested and plated.

[0306] Cell plating:

[0307] CT26 cells were trypsinized, harvested, and counted. Based on the count, the cells were diluted to 30,000 cells / mL in RPMI1640 + 10% FBS. The cells were then plated into 96-well flat-bottom culture plates, with 0.1 mL of cell suspension added to each well, representing 3,000 cells. After plating, the plates were placed in a 37°C, 5% CO2 incubator for continued culture.

[0308] Add the test compound:

[0309] 24 hours after plating the cells, the test compound, teniposide, was added to different wells of the cell plate. The test compounds were uniformly prepared and aliquoted prior to the experiment. Briefly, the drug was dissolved in DMSO to 10 mM, aliquoted into 50 μL vials, and stored at -20°C in the dark. For this experiment, one vial was removed for drug administration and treatment of the cells. Grouping and drug concentration information are shown in Table 8.

[0310] Table 8: Drug groups and dosing concentrations

[0311] Ten drug concentrations were designed for teniposide, the last of which was 0, which was set as the negative control group.

[0312] After drug addition, the drugs were gently mixed, and the cells were cultured in a 37°C, 5% CO2 incubator.

[0313] Add CCK-8 detection reagent and read the plate:

[0314] After 72 hours of drug exposure, 10 μL of CCK-8 assay reagent was added to each well of the cell plate using a multichannel pipette. The cell plate was then incubated for another 4 hours in a 37°C, 5% CO2 incubator. The absorbance of each well was measured at 450 nm using a microplate reader.

[0315] Data Analysis:

[0316] After the experiment, GraphPad Prism 8 software was used to analyze the inhibition percentage of the test drugs on the cells.

[0317] Calculation of inhibition percentage: Inhibition percentage = {[A(0 drug addition) - A(blank)] - [A(drug addition) - A(blank)]} / [A(0 drug addition) - A(blank)] × 100% A(drug addition): absorbance of wells with cells, CCK-8 solution, and drug solution A(blank): absorbance of wells with culture medium and CCK-8 solution but no cells A(0 drug addition): absorbance of wells with cells, CCK-8 solution, but no drug solution

[0318] Experimental conclusion:

[0319] This study evaluated the inhibitory effects of teniposide alone and in combination with IN10018 on CT26 cell proliferation in vitro. The relevant test results for each group 72 hours after drug treatment are shown in Figure 9. The IC50 value of the teniposide and IN10018 combination group was 0.09799 μM, lower than the IC50 value of the single-agent group (0.8179 μM). Therefore, the combination group has a stronger inhibitory effect on cancer cell growth in vitro.

[0320] Example 7: Study on the induction of immunogenic cell death targets by teniposide and IN10018 in mouse colon cancer CT26 cells in vitro

[0321] The experimental groups are shown in Table 9.

[0322] Table 9:

[0323] Compound information is shown in Table 10:

[0324] Table 10.

[0325] The compound information is shown in Table 11.

[0326] Table 11:

[0327] Cell culture

[0328] CT26 cells were maintained and passaged by Ins Biotech (Nanjing) Co., Ltd. Cells were cultured as monolayers in RPMI1640 medium supplemented with 10% fetal bovine serum at 37°C in a 5% CO2 incubator. Cells were routinely digested and passaged using trypsin two to three times a week. When cells reached 80%-90% confluence during the exponential growth phase, they were harvested and plated.

[0329] Cell plating

[0330] CT26 cells were digested with trypsin and then collected and counted. Based on the counting results, the cells were diluted with RPMI1640 + 10% FBS to a concentration of 50,000 cells per milliliter. The cells were then plated in a 12-well cell culture plate, with 2 ml of cell suspension, i.e., 100,000 cells, per well.

[0331] After plating, the cells were cultured in a 37°C, 5% CO2 incubator.

[0332] Add the test compound

[0333] 24 hours after plating, the test compound, teniposide, was added to different wells. The test compounds were uniformly prepared and aliquoted in DMSO at a concentration of 10 mM. Each aliquot was 50 μL per vial and stored at -20°C in the dark. For this experiment, one vial was removed for drug addition and testing. The groups and drug concentrations are shown in Table 9.

[0334] After 48 hours of drug treatment, the cells were photographed using a microscope, and then trypsinized and collected for flow cytometry staining.

[0335] After washing the cells twice with flow cytometry buffer (DPBS + 2% FBS), each group of cells was divided equally into two aliquots. 0.5 μL of AF647 Anti-Calreticulin Antibody was added to each well of one aliquot, mixed, and incubated at 4°C in the dark for 20 minutes. After washing once with flow cytometry buffer, 195 μL of Annexin-V-FITC conjugate was added. After gently mixing the cells, 5 μL of Annexin-V-FITC antibody was added. After further mixing, 10 μL of PI dye was added and mixed. The cells were incubated at room temperature in the dark for 15 minutes before flow cytometry analysis.

[0336] Data Analysis

[0337] After the experiment, Flowjo (V10) software was used to analyze the cell positive rate.

[0338] Experimental results:

[0339] This study evaluated the effects of teniposide alone and in combination with IN10018 on inducing the expression of immune cell death targets in CT26 cells under in vitro conditions.

[0340] The relevant test results for each group 48 hours after drug treatment are shown in Figure 10. Microscopic observation of cell status revealed significant cell death in the teniposide monotherapy group, with the most significant cell death in the combination therapy group. Cell viability was relatively good in the negative control group and IN10018 group. Flow cytometric analysis revealed that the CRT and Annexin V positivity rates in the combination therapy group were significantly higher than those in the monotherapy group.

[0341] Example 8: In vivo antitumor efficacy study of the combination of etoposide, IN10018, and anti-mouse PD-L1 antibody in a subcutaneous homograft model of colon cancer MC38 cells in C57BL / 6 mice

[0342] Grouping and dosing information are shown in Table 12.

[0343] Table 12: Animal groups and dosing regimens for in vivo efficacy experiments Note: 1. N: Number of mice per group; 2. Dosing volume: 10 mL / kg based on mouse body weight. If the body weight decreases by more than 15%, dosing should be stopped; dosing should be resumed after the body weight has recovered to 10% of the decrease. 3. The etoposide dose was increased to 6 mg / kg on the 6th day after group dosing.

[0344] Cell culture:

[0345] Colorectal cancer cell lines MC38 (source: Nanjing Kebai Biotechnology Co., Ltd., catalog number: CBP60825) were maintained and passaged by Ins Biotechnology (Nanjing) Co., Ltd. Cells were cultured as monolayers in DMEM supplemented with 10% fetal bovine serum at 37°C in a 5% CO2 incubator. Cells were routinely digested and passaged two to three times weekly using trypsin-EDTA. When cells were in the exponential growth phase and at 80%-90% saturation, they were harvested, counted, and plated.

[0346] Cell seeding and grouping:

[0347] 0.1 mL contains 2×10 5 A cell suspension of 100 cells was subcutaneously inoculated on the right back of each mouse. 3 Around 9 days after cell inoculation, the mice were randomly divided into groups according to tumor volume and given drugs. The preparations of the test substances and control solvents are shown in Table 13.

[0348] Table 13: Test substance and control solvent preparation

[0349] Daily observation of experimental animals:

[0350] The development of this experimental protocol and any modifications were reviewed and approved by the Yunqiao Biotech IACUC. The use and welfare of experimental animals were carried out in accordance with AAALAC guidelines. Animal health and mortality were monitored daily. Routine examinations included observation of tumor growth and the effects of drug treatment on the animals' daily behaviors, such as activity, food and water intake (visual observation only), weight changes, physical signs, or other abnormalities. Group deaths and adverse reactions were recorded based on the number of animals in each group.

[0351] Experiment termination:

[0352] If the animal's health condition continues to deteriorate, or the tumor volume exceeds 3,000 mm 3 If the animal is seriously ill or in pain, it must be euthanized. If the animal has any of the following conditions, notify the veterinarian and euthanize it: obvious weight loss, weight loss greater than 20%; cannot eat and drink freely; the average tumor volume of the control group reaches 3,000mm 3The experiment was terminated. The animal developed the following clinical manifestations and continued to worsen: piloerection, arched back, pale ears, nose, eyes, or feet, rapid breathing, convulsions, continuous diarrhea, dehydration, slow movement, and vocalization.

[0353] Tumor measurements and experimental parameters:

[0354] The experimental indicator is to examine whether tumor growth is inhibited, delayed, or cured. Tumor diameter is measured with a vernier caliper three times a week. The formula for calculating tumor volume is: V = 0.5 × a × b 2 , a and b represent the long diameter and short diameter of the tumor, respectively.

[0355] The tumor inhibition efficacy of the compound was evaluated using the TGI (%), which reflects the rate of tumor growth inhibition. The tumor growth inhibition rate (TGI) (%) was calculated using the following formula, referring to the tumor volume on the first day after grouping: TGI (%) = [1 - (average tumor volume of a given treatment group - average tumor volume of that treatment group at the start of treatment) / (average tumor volume of the solvent control group - average tumor volume of the solvent control group at the start of treatment)] × 100%.

[0356] Statistical analysis:

[0357] Statistical analysis was performed using Prism GraphPad software based on tumor volume at the end of the study. Multiple group comparisons were performed using two-way ANOVA and Fisher's LSD test. P < 0.05 was considered significant.

[0358] Experimental results:

[0359] 1. In vivo efficacy study of the combination of etoposide, IN10018, and anti-mouse PD-L1 antibody in a subcutaneous homograft model of MC38 colorectal cancer cells in C57BL / 6 mice

[0360] After cell inoculation, tumor growth was observed every day. Groups were divided according to tumor volume on the 9th day after inoculation. The average tumor volume in the group was about 50 mm 3 Due to tumor burden, the control group was euthanized on day 26 after inoculation, which was 17 days after group administration, and the entire experiment was terminated.

[0361] On the 17th day after group administration, the tumor volume of the control group was 2670.9±1438.9mm 3 The tumor volumes of the PD-L1 antibody (5 mg / kg) and etoposide (3 mg / kg) monotherapy groups were 2677.7±2029.0 mm 3 and 2420.2±1377.3mm 3The tumor volumes of the etoposide + PD-L1 antibody (3 + 5 mg / kg) and etoposide + IN10018 (3 + 25 mg / kg) combination treatment groups were 1947.6 ± 539.7 mm 3 and 1865.0±839.9mm 3 The tumor volumes of the etoposide + IN10018 + PD-L1 antibody (3 + 25 + 5 mg / kg) triplet treatment groups were 1245.6 ± 1064.7 mm 3 Compared with the control group, the tumor inhibition rates (TGI) of the PD-L1 antibody (5 mg / kg) and etoposide (3 mg / kg) monotherapy groups were -0.2% (p=0.9827) and 9.5% (p=0.4487), respectively. The tumor inhibition rates (TGI) of the etoposide + PD-L1 antibody (3+5 mg / kg) and etoposide + IN10018 (3+25 mg / kg) combination therapy groups were 27.6% (p=0.0295) and 30.7% (p=0.0111), respectively. The tumor inhibition rate (TGI) of the etoposide + IN10018 + PD-L1 antibody (3+25+5 mg / kg) triplet therapy group was 54.4% (p<0.0001).

[0362] Statistical analysis of the overall tumor volume was performed by comparing the control group with the etoposide + IN10018 + PD-L1 antibody (3 + 25 + 5 mg / kg) triplet combination group. The P values ​​for the control group, PD-L1 antibody (5 mg / kg), and etoposide (3 mg / kg) monotherapy groups were p < 0.0001, p < 0.0001, and p = 0.0005, respectively; the P values ​​for the etoposide + PD-L1 antibody (3 + 5 mg / kg) and etoposide + IN10018 (3 + 25 mg / kg) dual-drug combination treatment groups were p = 0.0346 and p = 0.0504, respectively. See Table 14 for details. The tumor volume of each dose group at different time periods is shown in Figure 11. Compared with the blank control group, the average tumor volume of the PD-L1 antibody (5 mg / kg) and etoposide (3 mg / kg) monotherapy groups was very close to that of the control group, and no inhibitory effect on tumor growth was shown; while the etoposide + PD-L1 antibody (3 + 5 mg / kg) and etoposide + IN10018 (3 + 25 mg / kg) combination therapy groups and the etoposide + IN10018 + PD-L1 antibody (3 + 25 + 5 mg / kg) triple therapy group all had significant tumor growth inhibitory effects, which were statistically different from the control group. Comprehensively considering the entire dosing cycle, the tumor volume of the etoposide + IN10018 + PD-L1 antibody (3+25+5 mg / kg) triple combination group was always smaller than that of the other treatment groups, and there were statistical differences compared with the PD-L1 antibody (5 mg / kg) and etoposide (3 mg / kg) monotherapy group and the etoposide + PD-L1 antibody (3+5 mg / kg) two-drug combination treatment group. Although there was no statistical difference between the etoposide + IN10018 + PD-L1 antibody (3+25+5 mg / kg) triple combination group and the etoposide + IN10018 (3+25 mg / kg) two-drug combination treatment group, their average tumor volume was relatively small, which also showed that the etoposide + IN10018 + PD-L1 antibody (3+25+5 mg / kg) triple combination group had a better effect in inhibiting tumor growth.

[0363] Table 14: Evaluation of the tumor inhibition effect of the test substance on the C57BL / 6 mouse transplant tumor model of mouse colon cancer MC38 cells (based on the data on the 17th day after group administration) Note: 1. Calculated according to the number of days after drug administration in each group, the data are mean ± standard deviation (mean ± SD); 2. TGI (%) = [1-(T 17 -T0) / (V 17-V0)] × 100%; 3.*: p < 0.05, vs. control group, two-way ANOVA; 4.*: p < 0.05, vs. etoposide + IN10018 + PD-L1 antibody (3 + 25 + 5 mg / kg) group, two-way ANOVA;

[0364] 2. Study on the effect of combination therapy of etoposide, IN10018, and anti-mouse PD-L1 antibody on body weight changes and clinical status in a C57BL / 6 mouse subcutaneous transplant model of MC38 colorectal cancer cells

[0365] The experiment was carried out according to the dosing regimen. During the experiment, the animals' activities such as eating and drinking were observed every day, and the weight of the animals was recorded three times a week. After 17 days of group administration, the average body weight of the control group changed from 18.7g on the day of group administration (Day 0) to 23.9g, with a body weight growth rate of 28.8%; the average body weight of the PD-L1 antibody (5mg / kg) and etoposide (3mg / kg) monotherapy groups changed from 18.8g and 19.1g on Day 0 to 23.6g and 22.8g on Day 17, respectively, with body weight change rates of 25.8% and 19.3%, respectively; The average body weight of the etoposide + IN10018 (3 + 25 mg / kg) combination treatment groups changed from 19.1 g and 18.9 g on Day 0 to 22.3 g and 22.5 g on Day 17, respectively, with body weight change rates of 17.2% and 19.0%, respectively; the average body weight of the etoposide + IN10018 + PD-L1 antibody (3 + 25 + 5 mg / kg) triplet combination group changed from 19.1 g on Day 0 to 21.6 g on Day 17, with a body weight change rate of 13.3%. During the entire dosing cycle, one mouse each in the etoposide (3 mg / kg) monotherapy group and the etoposide + PD-L1 antibody (3 + 5 mg / kg) combination therapy group died on day 12 and day 17 after group dosing due to tumor rupture. The animals in the remaining groups showed no significant weight loss and were in good condition throughout the dosing cycle, demonstrating tolerance to the different dosing methods. See Table 15 for details. The weight changes of each dose group at different time periods are shown in Figure 12. This indicates that the animals tolerated the triple drug combination of etoposide + IN10018 + PD-L1 antibody (3 + 25 + 5 mg / kg).

[0366] Table 15: Evaluation of the test substance on body weight changes in the C57BL / 6 mouse transplant tumor model of mouse colon cancer MC38 cells (based on data on day 17 after group administration) Note: 1. Calculated based on the number of days after group administration: number of animals surviving on day 0 / number of animals surviving on day 17; 2. Data are mean ± standard deviation (mean ± SD); 3. Body weight change rate = [1-(W 17 -W0) / W0]*100%;

[0367] Example 9: Study on the induction of immunogenic cell death targets by etoposide and AMP945 in mouse breast cancer 4T1 cells in vitro

[0368] Experimental Materials:

[0369] 1) Drugs used in this experiment

[0370] Etoposide provided by MCE, Lot No.: 113741

[0371] AMP945 provided by MCE, Lot No.:143253

[0372] 2) Antibodies used in this experiment

[0373] Restructuring Alexa 647 fluorescent anti-calreticulin antibody (Abcam, Cat No.: ab196159, Lot No.: CR33676773). Annexin V-apoptosis detection kit (Beyotime, Cat No.: C1062L, Lot No.: 122221220706).

[0374] Experimental methods:

[0375] 4T1 cells (source: Nanjing Kebai Biotechnology Co., Ltd., catalog number: CBP60352) were cultured in RPMI1640 (Shanghai Yuanpei, Cat No.: L210KJ, Lot No.: F210916) + 10% FBS (Gibco, Cat No.: 10099-141c, Lot No.: 2158737cp) at 37°C and 5% CO2. Two to three times a week, cells were routinely digested and passaged using trypsin. When the cells were in the exponential growth phase and 80%-90% confluent, they were harvested and plated. After trypsinization, the 4T1 cells were harvested and counted. Based on the count results, the cells were diluted in RPMI1640 + 10% FBS to a concentration of 50,000 cells / mL. The cells were then plated in 12-well cell culture plates, with 2 mL of cell suspension per well, i.e., 100,000 cells. After plating, the cells were cultured in a 37°C, 5% CO2 incubator. After 24 hours of cell spreading, six groups were set up: the first group served as the control group and received culture medium; the second group received AMP945 at a concentration of 3 μM; the third group received AMP945 at a concentration of 6 μM; the fourth group received etoposide at a concentration of 5 μM; the fifth group received a combination of AMP945 (3 μM) and etoposide (5 μM); and the sixth group received a combination of AMP945 (6 μM) and etoposide (5 μM). The drugs were mixed and cultured at 37°C in a 5% CO2 incubator for 48 hours.

[0376] Experimental results

[0377] After 48 hours of drug exposure, cells were collected for flow cytometry analysis and washed twice with flow cytometry buffer (PBS + 2% FBS). 0.5 μl of AF647 anti-calreticulin antibody (abcam) was added to each well and mixed. The cells were incubated at 4°C in the dark for 20 minutes. After that, flow cytometry buffer was added. Using the Annexin V-Apoptosis Detection Kit (Beyotime), 195 μl of Annexin-V-FITC conjugate was added and mixed by pipetting. 5 μl of Annexin-V-FITC antibody was added and gently mixed. Finally, 10 μl of PI dye was added and mixed. The mixture was incubated at room temperature in the dark for 15 minutes. The samples were then transferred to a flow cytometer for signal measurement.

[0378] The results of flow cytometry analysis showed that the CRT positivity rate and Annexin-V positivity rate of the combination group were significantly better than those of the single-drug group and the control group, as shown in Figure 13.

[0379] Example 10: Study on the induction of immunogenic cell death targets by irinotecan and AMP945 in mouse breast cancer 4T1 cells in vitro

[0380] Experimental Materials:

[0381] 1) Drugs used in this experiment

[0382] Irinotecan provided by Shanghai Chaolan Chemical Technology Center, Lot No.: 202110

[0383] AMP945 provided by MCE, Lot No.:143253

[0384] 2) Antibodies used in this experiment

[0385] Restructuring Alexa 647 fluorescent anti-calreticulin antibody (Abcam, Cat No.: ab196159, Lot No.: CR33676773). Annexin V-apoptosis detection kit (Beyotime, Cat No.: C1062L, Lot No.: 122221220706).

[0386] Experimental methods:

[0387] 4T1 cells (source: Nanjing Kebai Biotechnology Co., Ltd., catalog number: CBP60352) were cultured in RPMI1640 (Shanghai Yuanpei, Cat No.: L210KJ, Lot No.: F210916) + 10% FBS (Gibco, Cat No.: 10099-141c, Lot No.: 2158737cp) at 37°C and 5% CO2. Two to three times a week, cells were routinely digested and passaged using trypsin. When the cells were in the exponential growth phase and 80%-90% confluent, they were harvested and plated. After trypsinization, the 4T1 cells were harvested and counted. Based on the count results, the cells were diluted in RPMI1640 + 10% FBS to a concentration of 50,000 cells / mL. The cells were then plated in 12-well cell culture plates, with 2 mL of cell suspension per well, i.e., 100,000 cells. After plating, the cells were cultured in a 37°C, 5% CO2 incubator. After 24 hours of cell spreading, six groups were set up: the first group was a control group, supplemented with culture medium; the second group was treated with AMP945 at a concentration of 3 μM; the third group was treated with AMP945 at a concentration of 6 μM; the fourth group was treated with irinotecan at a concentration of 120 μM; the fifth group was treated with AMP945 (3 μM) combined with irinotecan (120 μM); and the sixth group was treated with AMP945 (6 μM) combined with irinotecan (120 μM). The drugs were mixed and cultured at 37°C in a 5% CO2 incubator for 48 hours.

[0388] Experimental results

[0389] After 48 hours of drug exposure, cells were collected for flow cytometry analysis and washed twice with flow cytometry buffer (PBS + 2% FBS). 0.5 μl of AF647 anti-calreticulin antibody (abcam) was added to each well and mixed. The cells were incubated at 4°C in the dark for 20 minutes. After that, flow cytometry buffer was added. Using the Annexin V-Apoptosis Detection Kit (Beyotime), 195 μl of Annexin-V-FITC conjugate was added and mixed by pipetting. 5 μl of Annexin-V-FITC antibody was added and gently mixed. Finally, 10 μl of PI dye was added and mixed. The mixture was incubated at room temperature in the dark for 15 minutes. The samples were then transferred to a flow cytometer for signal measurement.

[0390] The results of flow cytometry analysis showed that the CRT positivity rate and Annexin-V positivity rate of the combination group were significantly better than those of the single-drug group and the control group, as shown in Figure 14.

[0391] All references mentioned in this application are incorporated herein by reference in their entirety, just as if each reference were listed separately. It should be understood that after reading the disclosure of this application, those skilled in the art may make various changes or modifications to the present invention, and these equivalent forms also fall within the scope of the claims appended hereto.

Claims

1. Use of a FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor in the preparation of a medicament for treating a tumor in a subject, wherein the topoisomerase inhibitor is not an anthracycline.

2. A pharmaceutical combination product of a FAK inhibitor, a topoisomerase inhibitor and an immune checkpoint inhibitor for use in treating a tumor in a subject, wherein the topoisomerase inhibitor is not an anthracycline.

3. A method for treating a tumor, comprising administering to a subject a therapeutically effective amount of a FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

4. The use, pharmaceutical combination or method of any one of claims 1 to 3, wherein the FAK inhibitor and the topoisomerase inhibitor induce immunogenic cell death (ICD), wherein the topoisomerase inhibitor is not an anthracycline.

5. The use, pharmaceutical combination or method according to any one of claims 1 to 4, wherein the FAK inhibitor is IN10018, Defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886 or a pharmaceutically acceptable salt thereof, preferably IN10018, Defactinib, AMP945 or a pharmaceutically acceptable salt thereof, further preferably IN10018 or a pharmaceutically acceptable salt thereof, especially IN10018 tartrate, and the structure of IN10018 is as follows:

6. The use, pharmaceutical combination or method according to any one of claims 1 to 5, wherein the topoisomerase inhibitor is a topoisomerase I inhibitor.

7. The use, pharmaceutical combination or method of claim 6, wherein the topoisomerase I inhibitor is topotecan, irinotecan, belotecan, cositecan, exatecan (DX-8951), indenoisoquinoline, indotecan (LMP-400), indimitecan (LMP-776), simmitecan, gimatecan (ST1481), EC-112002, PLX-038 (NK012), AR-67, or a pharmaceutically acceptable salt thereof.

8. The use, pharmaceutical combination or method of claim 7, wherein the topoisomerase I inhibitor is irinotecan, or a pharmaceutically acceptable salt thereof.

9. The use, pharmaceutical combination or method according to any one of claims 1 to 5, wherein the topoisomerase inhibitor is a topoisomerase II inhibitor.

10. The use, pharmaceutical combination or method of claim 9, wherein the topoisomerase II inhibitor is etoposide, teniposide, sabarubicin, or a pharmaceutically acceptable salt thereof.

11. The use, pharmaceutical combination or method of claim 10, wherein the topoisomerase II inhibitor is etoposide, teniposide, or a pharmaceutically acceptable salt thereof.

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

13. The use, pharmaceutical combination or method according to any one of claims 1 to 12, wherein 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, atezolizumab, camrelizumab, sintilimab, cemiplimab, envafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.

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

15. The use, pharmaceutical combination product or method according to any one of claims 1 to 12, wherein the immune checkpoint inhibitor is a TIGIT antibody, and further, the TIGIT antibody is Ociperlimab (BGB-A1217), Vibostolimab, domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.

16. The use, pharmaceutical combination product or method according to any one of claims 1 to 5, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; the topoisomerase inhibitor is irinotecan 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, in particular an anti-PD-1 / PD-L1 antibody.

17. The use, pharmaceutical combination product or method according to any one of claims 1 to 5, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; the topoisomerase inhibitor is etoposide, teniposide, 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, especially an anti-PD-1 / PD-L1 antibody.

18. The use, pharmaceutical combination or method of any one of claims 1 to 17, wherein the FAK inhibitor, the topoisomerase inhibitor and the immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.

19. The use, pharmaceutical combination or method of any one of claims 1 to 18, wherein 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, bile duct cancer, leiomyosarcoma, liposarcoma, nasopharyngeal carcinoma, neuroendocrine cancer, The invention relates to a method for treating a thyroid cancer of the present invention and the like. The method comprises the following steps: treating a thyroid cancer of the present invention and the like; wherein the thyroid cancer is a thyroid cancer, a thyroid cancer, a salivary gland cancer, a metastatic tumor caused by spindle cell carcinoma, an anaplastic large cell lymphoma, an undifferentiated thyroid cancer, a non-Hodgkin's lymphoma, a Hodgkin's lymphoma, a 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), chronic myeloid leukemia (CML); preferably, 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. The use, pharmaceutical combination or method of any one of claims 1 to 19, wherein the tumor is breast cancer or colon cancer (including colorectal cancer).

21. A kit or pharmaceutically acceptable composition comprising: (a) FAK inhibitors; (b) a topoisomerase inhibitor, wherein the topoisomerase inhibitor is not an anthracycline; and (c) Immune checkpoint inhibitors.

22. The kit or composition of claim 21, wherein the FAK inhibitor and the topoisomerase inhibitor induce immunogenic cell death (ICD).

23. The kit or composition according to any one of claims 21 to 22, wherein the FAK inhibitor is IN10018, Defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886, or a pharmaceutically acceptable salt thereof, preferably IN10018, Defactinib, AMP945, or a pharmaceutically acceptable salt thereof, further preferably IN10018 or a pharmaceutically acceptable salt thereof, especially IN10018 tartrate, wherein the structure of IN10018 is as follows:

24. The kit or composition of any one of claims 21 to 23, wherein the topoisomerase inhibitor is a topoisomerase I inhibitor.

25. The kit or composition of claim 24, wherein the topoisomerase I inhibitor is topotecan, irinotecan, belotecan, cositecan, exatecan (DX-8951), indenoisoquinoline, indotecan (LMP-400), indimitecan (LMP-776), simmitecan, gimatecan (ST1481), EC-112002, PLX-038 (NK012), AR-67, or a pharmaceutically acceptable salt thereof.

26. The kit or composition of claim 25, wherein the topoisomerase I inhibitor is irinotecan, or a pharmaceutically acceptable salt thereof.

27. The kit or composition of any one of claims 21-23, wherein the topoisomerase inhibitor is a topoisomerase II inhibitor.

28. The kit or composition of claim 27, wherein the topoisomerase II inhibitor is etoposide, teniposide, sabarubicin, or a pharmaceutically acceptable salt thereof.

29. The kit or composition of claim 28, wherein the topoisomerase II inhibitor is etoposide, teniposide, or a pharmaceutically acceptable salt thereof.

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

31. The kit or composition of any one of claims 21 to 30, wherein 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, Atezolizumab, Camrelizumab, Sintilimab, Cemiplimab, envafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005, or MX-10181.

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

33. The kit or composition of any one of claims 21-30, wherein the immune checkpoint inhibitor is a TIGIT antibody, and further, the TIGIT antibody is Ociperlimab / BGB-A1217, Vibostolimab, domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.

34. The kit or composition of any one of claims 21-22, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; the topoisomerase inhibitor is irinotecan 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, especially an anti-PD-1 / PD-L1 antibody.

35. The kit or composition of any one of claims 21-22, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; the topoisomerase inhibitor is etoposide, teniposide, or a pharmaceutically acceptable salt thereof; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, especially an anti-PD-1 / PD-L1 antibody.

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

37. The kit or composition of any one of claims 21 to 36, wherein the drug is used to treat a tumor, and 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, bile duct cancer, leiomyosarcoma, liposarcoma, nasopharyngeal carcinoma, neuroblastoma, Metastases caused by endocrine cancer, ovarian cancer, salivary gland cancer, spindle cell cancer, anaplastic large cell lymphoma, undifferentiated thyroid cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma, glioma or hematological malignancies, such as acute myeloid leukemia (AML), acute lymphocytic leukemia (ALL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML); preferably, 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. The kit or composition of claim 37, wherein the tumor is breast cancer or colon cancer (including colorectal cancer).

39. A FAK inhibitor for use in the treatment of tumors for enhancing immunogenic cell death induced by a topoisomerase inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.

40. The FAK inhibitor according to claim 39, wherein the FAK inhibitor is IN10018, Defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886, or a pharmaceutically acceptable salt thereof, preferably IN10018, Defactinib, AMP945, or a pharmaceutically acceptable salt thereof, further preferably IN10018 or a pharmaceutically acceptable salt thereof, especially IN10018 tartrate, and the structure of IN10018 is as follows:

41. The FAK inhibitor of claim 39 or 40, wherein the topoisomerase inhibitor is a topoisomerase I inhibitor.

42. The FAK inhibitor of claim 41, wherein the topoisomerase I inhibitor is topotecan, irinotecan, belotecan, cositecan, exatecan (DX-8951), indenoisoquinoline, indotecan (LMP-400), indimitecan (LMP-776), simmitecan, gimatecan (ST1481), EC-112002, PLX-038 (NK012), AR-67, or a pharmaceutically acceptable salt thereof.

43. The FAK inhibitor according to claim 42, wherein the topoisomerase I inhibitor is irinotecan, or a pharmaceutically acceptable salt thereof.

44. The FAK inhibitor of claim 39 or 40, wherein the topoisomerase inhibitor is a topoisomerase II inhibitor.

45. The FAK inhibitor according to claim 44, wherein the topoisomerase II inhibitor is etoposide, teniposide, sabarubicin, or a pharmaceutically acceptable salt thereof. The FAK inhibitor according to claim 45 , wherein the topoisomerase II inhibitor is etoposide, teniposide, or a pharmaceutically acceptable salt thereof.

47. The FAK inhibitor of any one of claims 39-46, wherein 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, bile duct cancer, leiomyosarcoma, liposarcoma, nasopharyngeal carcinoma, neuroendocrine cancer, Ovarian cancer, salivary gland cancer, metastases caused by spindle cell carcinoma, anaplastic large cell lymphoma, undifferentiated thyroid cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma, glioma or hematological malignancies, such as acute myeloid leukemia (AML), acute lymphocytic leukemia (ALL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML); preferably, 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.

48. The FAK inhibitor according to any one of claims 39 to 47, wherein the tumor is breast cancer or colon cancer (including colorectal cancer).

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