Pharmaceutical composition of FAK inhibitor and microtubule inhibitor and application thereof

Abstract

The invention relates to a composition of an FAK inhibitor, a microtubule inhibitor and an immune checkpoint inhibitor, a kit composed of the composition and application of the composition in preparation of drugs for treating tumors.

Description

Drug combination and use of FAK inhibitor and microtubule inhibitor

[0001] This application claims priority to Chinese patent application No. 202211018821.9 filed on August 24, 2022, Chinese patent application No. 202211165722.3 filed on September 23, 2022, and Chinese patent application No. 202310940775.6 filed on July 28, 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 cause of death and a leading cause of death. Immunogenic cell death (ICD) is a cascade of effects that occur in programmed cell death (PND). Exposure of cancer cells to chemotherapy or targeted therapies may activate internal stress signals, including endoplasmic reticulum (ER) stress and reactive oxygen species (ROS) stress. In response to these stress signals, cells first attempt to repair the damage. If the damage caused by stress exceeds their ability to repair, they initiate programmed cell death. This process is often accompanied by the release of damage-associated molecular patterns (DAMPs), including calreticulin, annexin A1, and HMGB1. These DAMPs are specifically recognized by pattern recognition receptors on antigen-presenting cells (APCs), inducing APC maturation, differentiation, and activation. These DAMPs are then presented to immune cells, including effector T cells, thereby generating antigen memory in these immune cells. 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] Microtubules are dynamic, filamentous cytoskeletal proteins and the main component of centrioles. The orderly dynamic changes of microtubule fibers are crucial for cell mitosis. Microtubule inhibitors bind to tubulin to promote or inhibit microtubule assembly, disrupting the normal structure and function of the microtubule-forming spindle. This has the potential to interfere with cell mitosis and inhibit cell proliferation, exerting a clear anti-tumor effect. Anti-tumor drugs targeting microtubules have become an important aspect of current anti-tumor drug research. Many microtubule inhibitors are currently on the market, including taxanes and eribulin. Taxanes include paclitaxel, a drug extracted from natural tree bark, as well as synthetic docetaxel and, now, albumin-paclitaxel. Taxanes are M-phase-specific drugs that promote the polymerization of tubules into stable microtubules and inhibit their dissociation, significantly reducing the number of tubules and disrupting the microtubule network. Eribulin is a synthetic halichondrin B analogue that inhibits microtubule depolymerization, leading to G2 / M arrest in the cell cycle and disruption of the mitotic spindle, ultimately causing cell apoptosis. Taxanes and eribulin are both commonly used chemotherapy drugs in clinical practice. However, both can develop resistance after a period of use.

[0006] Therefore, there is still a need to find a way to improve the efficacy of single drugs in chemotherapy and further overcome the problem of drug resistance.

[0007] Summary of the Invention

[0008] In one aspect, the present disclosure provides uses of a FAK inhibitor, a microtubule inhibitor, and an immune checkpoint inhibitor in the preparation of a medicament for treating a tumor in a subject.

[0009] Yet another aspect of the present disclosure provides a pharmaceutical combination product of a FAK inhibitor, a microtubule inhibitor, and an immune checkpoint inhibitor for use in treating a tumor in a subject.

[0010] In another aspect, the present disclosure provides a method for treating tumors, comprising administering a therapeutically effective amount of a FAK inhibitor, a microtubule inhibitor, and an immune checkpoint inhibitor to a subject in need thereof.

[0011] In another aspect, the present disclosure provides a kit or a pharmaceutically acceptable composition comprising: (a) a FAK inhibitor; (b) a microtubule inhibitor; and (c) an immune checkpoint inhibitor.

[0012] In another aspect, the present disclosure provides use of a FAK inhibitor and a microtubule inhibitor in preparing a medicament for treating tumors, wherein the FAK inhibitor is used to enhance the immunogenic cell death induced by the microtubule inhibitor.

[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 microtubule inhibitors.

[0014] Yet another aspect of the present disclosure provides a method for treating tumors, comprising administering a therapeutically effective amount of a FAK inhibitor and a microtubule inhibitor to a subject in need thereof, wherein the FAK inhibitor is used to enhance immunogenic cell death induced by the microtubule inhibitor.

[0015] In another aspect, the present disclosure provides the use of FAK inhibitors, microtubule inhibitors and immune checkpoint inhibitors in the preparation of drugs for combined treatment of tumors.

[0016] In another aspect, the present disclosure provides use of a FAK inhibitor in the preparation of a drug for use in combination with a microtubule inhibitor and an immune checkpoint inhibitor in treating tumors.

[0017] In another aspect, the present disclosure provides use of a microtubule inhibitor in the preparation of a drug for use in combination with a FAK inhibitor and an immune checkpoint inhibitor in treating tumors.

[0018] In another aspect, the present disclosure provides the use of an immune checkpoint inhibitor in the preparation of a drug for use in combination with a FAK inhibitor and a microtubule inhibitor in treating tumors.

[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 microtubule inhibitor and an immune checkpoint inhibitor.

[0020] In another aspect, the present disclosure provides use of a microtubule inhibitor in the preparation of a medicament for treating tumors in combination with a FAK inhibitor and an immune checkpoint inhibitor.

[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 microtubule inhibitor.

[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 microtubule inhibitor and an immune checkpoint inhibitor to treat tumors.

[0023] In another aspect, the present disclosure provides a kit comprising: a microtubule inhibitor; and instructions, wherein the instructions indicate that the microtubule inhibitor can be used in combination with a FAK inhibitor and an immune checkpoint inhibitor to treat tumors.

[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 microtubule inhibitor to treat tumors.

[0025] Another aspect of the present disclosure provides a method for treating a tumor, comprising administering a therapeutically effective amount of a FAK inhibitor and a microtubule inhibitor to a subject in need thereof.

[0026] Yet another aspect of the present disclosure provides a pharmaceutical combination product of a FAK inhibitor and a microtubule inhibitor for use in treating a tumor in a subject in need thereof.

[0027] In another aspect, the present disclosure provides use of a FAK inhibitor and a microtubule inhibitor in preparing a combined drug for treating tumors.

[0028] In another aspect, the present disclosure provides use of a FAK inhibitor in preparing a medicament for use in combination with a microtubule inhibitor in treating tumors.

[0029] In another aspect, the present disclosure provides use of a microtubule inhibitor in preparing a medicament for use in combination with a FAK inhibitor in treating tumors.

[0030] In another aspect, the present disclosure provides use of a FAK inhibitor and a microtubule inhibitor in the preparation of a medicament for combined treatment of tumors.

[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 microtubule inhibitor.

[0032] In another aspect, the present disclosure provides use of a microtubule inhibitor in preparing a medicament for treating tumors in combination with a FAK inhibitor.

[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 microtubule inhibitor to treat tumors.

[0034] In another aspect, the present disclosure provides a kit comprising: a microtubule inhibitor; and instructions, wherein the instructions indicate that the microtubule inhibitor can be used in combination with a FAK inhibitor to treat tumors.

[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 microtubule inhibitor is a taxane, Eribulin, Ixempra or vinca alkaloid.

[0038] Optionally, the microtubule inhibitor is a taxane.

[0039] Optionally, the taxane is docetaxel, paclitaxel, cabazitaxel or cephalomannine, preferably docetaxel or paclitaxel.

[0040] Optionally, the microtubule inhibitor is docetaxel.

[0041] Optionally, the microtubule inhibitor is paclitaxel.

[0042] The paclitaxel or docetaxel includes preparations thereof, such as liposome paclitaxel, albumin paclitaxel, and the like.

[0043] Optionally, the microtubule inhibitor is Eribulin, which is also called Eribulin and has a CAS number of 253128-41-5.

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

[0045] Optionally, the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, and further, the anti-PD-1 / PD-L1 antibody is pembrolizumab, Tislelizumab, Nivolumab, Toripalimab, Atezolizumab, durvalumab, Avelumab, Camrelizumab, Sintilimab, Cemiplimab, envafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.

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

[0047] Optionally, the immune checkpoint inhibitor is a TIGIT inhibitor, and further, the TIGIT inhibitor 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.

[0048] Optionally, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is docetaxel; and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor.

[0049] Optionally, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is docetaxel; and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

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

[0051] Optionally, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is paclitaxel; and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0052] Optionally, the FAK inhibitor, the microtubule inhibitor and the immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.

[0053] 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).

[0054] Optionally, the tumor is preferably breast cancer, ovarian cancer, colon cancer (including colorectal cancer), lung cancer (including small cell lung cancer and non-small cell lung cancer), melanoma or pancreatic cancer.

[0055] Optionally, the tumor is breast cancer, ovarian 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 FAK silencing combined with 0.3 μM or 1 μM Docetaxel for 48 hours, followed by staining using the Annexin V kit.

[0058] FIG2 shows that FAK silencing combined with 1 μM Docetaxel for 48 hours enhanced calreticulin release and exposure.

[0059] Figure 3 shows FAK silencing combined with 0.15 μM Eribulin for 48 hours, followed by staining using the Annexin V kit.

[0060] FIG4 shows that FAK silencing combined with 0.15 μM Eribulin for 48 hours enhanced the release and exposure of calreticulin.

[0061] FIG5 shows the IC50 values ​​of Docetaxel 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] FIG7 a shows the percentage of CRT-positive cells after colon cancer CT26 cells were incubated with drugs for 48 hours; FIG7 b shows the percentage of Annexin V-positive cells after colon cancer CT26 cells were incubated with drugs for 48 hours.

[0064] Figure 8a shows the percentage of Calreticulin (CRT)-positive cells in mouse breast cancer 4T1 cells after incubation with drugs for 48 hours; Figure 8b shows the percentage of Annexin-V-positive cells in mouse breast cancer 4T1 cells after incubation with drugs for 48 hours; Figure 8c shows the percentage of GRP94-positive cells in mouse breast cancer 4T1 cells after incubation with drugs for 48 hours.

[0065] FIG9 a shows the percentage of Calreticulin (CRT)-positive cells in mouse ovarian epithelial cancer ID8 cells after incubation with drugs for 48 hours; FIG9 b shows the percentage of Annexin-V-positive cells in mouse ovarian epithelial cancer ID8 cells after incubation with drugs for 48 hours.

[0066] Figure 10a shows the percentage of Calreticulin (CRT)-positive cells in human ovarian epithelial cancer TOV-21G cells after incubation with drugs for 48 hours; Figure 10b shows the percentage of Annexin-V-positive cells in human ovarian epithelial cancer TOV-21G cells after incubation with drugs for 48 hours.

[0067] Figure 11a shows the percentage of Calreticulin (CRT)-positive cells in mouse colon cancer CT26 cells after incubation with drugs such as Docetaxel for 48 hours; Figure 11b shows the percentage of Annexin-V-positive cells in mouse colon cancer CT26 cells after incubation with drugs such as Docetaxel for 48 hours.

[0068] Figure 12a shows the percentage of Calreticulin (CRT)-positive cells in mouse colon cancer CT26 cells after incubation with Eribulin and other drugs for 48 hours; Figure 12b shows the percentage of Annexin-V-positive cells in mouse colon cancer CT26 cells after incubation with Eribulin and other drugs for 48 hours.

[0069] Figure 13a shows the percentage of Calreticulin (CRT)-positive cells in mouse colon cancer CT26 cells after incubation with Paclitaxel and other drugs for 48 hours; Figure 13b shows the percentage of Annexin-V-positive cells in mouse colon cancer CT26 cells after incubation with Paclitaxel and other drugs for 48 hours.

[0070] Figure 14a shows the percentage of Calreticulin (CRT)-positive cells in mouse breast cancer 4T1 cells after incubation with Docetaxel and other drugs for 48 hours; Figure 14b shows the percentage of Annexin-V-positive cells in mouse breast cancer 4T1 cells after incubation with Docetaxel and other drugs for 48 hours.

[0071] FIG15 a shows the percentage of Calreticulin (CRT)-positive cells in mouse breast cancer 4T1 cells after incubation with Eribulin and other drugs for 48 hours; FIG15 b shows the percentage of Annexin-V-positive cells in mouse breast cancer 4T1 cells after incubation with Eribulin and other drugs for 48 hours.

[0072] FIG16 shows the changes in tumor volume after administration of different test substances in a BALB / c mouse subcutaneous transplant tumor model of breast cancer 4T1 cells.

[0073] FIG17 shows the changes in mouse body weight after administration of different test substances in a BALB / c mouse subcutaneous transplant tumor model of breast cancer 4T1 cells.

[0074] Figure 18a shows the percentage of Calreticulin (CRT)-positive cells in mouse breast cancer 4T1 cells after incubation with Docetaxel and other drugs for 48 hours; Figure 18b shows the percentage of Annexin-V-positive cells in mouse breast cancer 4T1 cells after incubation with Docetaxel and other drugs for 48 hours.

[0075] FIG19a shows the percentage of Calreticulin (CRT)-positive cells in mouse breast cancer 4T1 cells after incubation with Eribulin and other drugs for 48 hours; FIG19b shows the percentage of Annexin-V-positive cells in mouse breast cancer 4T1 cells after incubation with Eribulin and other drugs for 48 hours. DETAILED DESCRIPTION

[0076] 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.

[0077] 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.

[0078] 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.

[0079] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0080] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as 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.

[0081] 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.

[0082] 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."

[0083] definition

[0084] 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, APG-2449, AMP945, AMP886, or a pharmaceutically acceptable salt thereof, and the structure of IN10018 is as follows:

[0085] 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, or a pharmaceutically acceptable salt thereof. In some preferred embodiments, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, particularly IN10018 tartrate.

[0086] The term "microtubule inhibitor" as used herein refers to an effective inhibitor of microtubules / tubulin, which can be suitable for mammals, especially humans. Microtubules are components of the cytoskeleton and are distributed throughout the cytoplasm. Tubulin is one of several members of the small globin family. The tubulin superfamily includes five different families, α, β, γ, δ and ε tubulin, and the sixth family is only found in animal and plant embryonic protozoa. The most common members of the tubulin family are α-tubulin and β-tubulin, which are proteins that make up microtubules. Microtubule inhibitors bind to tubulin to promote or inhibit the assembly of microtubules, interfere with the normal structure and function of the spindle composed of microtubules, have the effect of interfering with cell mitosis and inhibiting cell proliferation, and exert a clear anti-tumor effect. Microtubule inhibitors are a large class of chemotherapy drugs, including taxanes, eribulin, ixabepilone and vinca alkaloids.

[0087] As used herein, the term "taxanes" refers to a broad class of chemotherapy drugs, including paclitaxel, a drug extracted from natural tree bark, and the synthetic docetaxel. For the avoidance of doubt, references to paclitaxel or docetaxel in the claims of this invention include their formulations, such as liposome paclitaxel and albumin paclitaxel.

[0088] The eribulin is also called Eribulin, and its CAS number is 253128-41-5.

[0089] The ixabepilone is also known as Ixabepilone, azaepothilone B or BMS-247550, and its CAS number is 219989-84-1.

[0090] 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 (Nivolumab), toripalimab (Toripalimab / Tuoyi), durvalumab (Improvin / 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, the PD-1 inhibitor is used to treat a human subject. In some embodiments, the PD-1 is human PD-1.

[0091] PD-1 / PD-L1 inhibitors also include PD-1 / PD-L1 small molecule inhibitors. In some embodiments, the PD-1 / PD-L1 small molecule inhibitor is INCB-086550, Lazertinib, IMH-010, CA-170, ABSK043, or RRx-001.

[0092] TIGIT (also known as WUCAM, Vstm3, and VSIG9) is a receptor of the Ig superfamily and a new immune checkpoint after PD-1 / PD-L1. For example, in the treatment methods, drugs and uses disclosed herein, the PD-1 / PD-L1 inhibitor is a TIGIT inhibitor, and the TIGIT inhibitor includes but is 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, TIGIT inhibitors are used to treat human subjects. For the avoidance of doubt, antibodies herein include bispecific antibodies.

[0093] 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.

[0094] 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.

[0095] "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.

[0096] 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.

[0097] 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.

[0098] 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.

[0099] 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.

[0100] 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 preferably breast cancer, ovarian cancer, colon cancer (including colorectal cancer), lung cancer (including small cell lung cancer and non-small cell lung cancer), melanoma or pancreatic cancer; in some embodiments, the tumor is breast cancer, ovarian cancer or colon cancer (including colorectal cancer).

[0101] 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.

[0102] As used herein, the term "pharmaceutically acceptable" means non-toxic, biologically tolerable, and suitable for administration to a subject.

[0103] 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.

[0104] 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.

[0105] 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.).

[0106] As used herein, the term "inhibit" refers to a decrease in the baseline activity of a biological activity or process.

[0107] 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 microtubule inhibitor, and an immune checkpoint inhibitor; and (ii) instructions, the instructions indicating that FAK inhibitors, microtubule 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, microtubule inhibitors, and immune checkpoint inhibitors can be used to treat tumors in a subject. In one embodiment, the kit includes (i) a microtubule inhibitor; and (ii) instructions, the instructions indicating that FAK inhibitors, microtubule 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, microtubule 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 microtubule inhibitor, and an immune checkpoint inhibitor; and (ii) instructions indicating that the FAK inhibitor, microtubule inhibitor, and immune checkpoint inhibitor can be used to treat tumors in a subject. In one embodiment, the kit includes (i) a FAK inhibitor; and (ii) instructions indicating that the FAK inhibitor and microtubule inhibitor can be used to treat tumors in a subject. In one embodiment, the kit includes (i) a microtubule inhibitor; and (ii) instructions indicating that the microtubule inhibitor and FAK inhibitor can be used to treat tumors in a subject.

[0108] 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 a medicine including a FAK inhibitor, the second container includes at least one dose of a microtubule inhibitor, the third container includes at least one dose of an immune checkpoint inhibitor, and the package insert includes instructions for using a drug to treat the tumor of an 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.

[0109] The precise amount of FAK inhibitor, microtubule 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.

[0110] The FAK inhibitor, microtubule inhibitor, or immune checkpoint inhibitor can be administered by a suitable route, such as oral, intravenous, intramuscular, or subcutaneous administration.

[0111] 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.

[0112] 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.

[0113] 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.

[0114] 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.

[0115] The amount of FAK inhibitor, microtubule 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.

[0116] 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. In a specific embodiment, 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 the free base.

[0117] The microtubule inhibitor is administered at a dose of 20-60 mg / m3 per week in adults. 2 In a specific embodiment, docetaxel is administered at a dose range of 20-25 mg / m2 per week in adults. 2 Paclitaxel or its preparations are administered at a dose of 45-60 mg / m per week in adults 2 The dosage is based on paclitaxel; Eribulin is administered at 1.4 mg / m 2 Administer intravenously over 2 to 5 minutes on Days 1 and 8 of a 21-day course of treatment.

[0118] 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.

[0119] 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.

[0120] In some embodiments, the present disclosure further discloses the following:

[0121] 1. A FAK inhibitor, a microtubule inhibitor, and an immune checkpoint inhibitor for use in a method of treating a tumor in a subject.

[0122] 2. The FAK inhibitor, microtubule 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:

[0123] 3. The FAK inhibitor, microtubule inhibitor, and immune checkpoint inhibitor according to embodiments 1-2, wherein the microtubule inhibitor is a taxane, eribulin, ixabepilone, or vinca alkaloid.

[0124] 4. The FAK inhibitor, microtubule inhibitor, and immune checkpoint inhibitor according to any one of embodiments 1-3, wherein the microtubule inhibitor is a taxane.

[0125] 5. The FAK inhibitor, microtubule inhibitor and immune checkpoint inhibitor according to any one of embodiments 3-4, wherein the taxane is docetaxel, paclitaxel, cabazitaxel or cephalomannine, preferably docetaxel or paclitaxel.

[0126] 6. The FAK inhibitor, microtubule inhibitor and immune checkpoint inhibitor according to any one of embodiments 1-5, wherein the microtubule inhibitor is docetaxel.

[0127] 7. The FAK inhibitor, microtubule inhibitor and immune checkpoint inhibitor according to any one of embodiments 1-5, wherein the microtubule inhibitor is paclitaxel.

[0128] 8. The FAK inhibitor, microtubule inhibitor, and immune checkpoint inhibitor according to any one of embodiments 1-4, wherein the microtubule inhibitor is Eribulin.

[0129] 9. The FAK inhibitor, microtubule 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 inhibitor.

[0130] 10. The FAK inhibitor, microtubule 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, durvalumab, Avelumab, Camrelizumab, Sintilimab, Cemiplimab, envafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005, or MX-10181.

[0131] 11. The FAK inhibitor, microtubule inhibitor, and immune checkpoint inhibitor according to any one of embodiments 1-9, 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.

[0132] 12. The FAK inhibitor, microtubule inhibitor and immune checkpoint inhibitor according to any one of embodiments 1 to 9, wherein the immune checkpoint inhibitor is a TIGIT inhibitor, and further, the TIGIT inhibitor is Ociperlimab (Ociperlimab / BGB-A1217), Vibostolimab, domvanalimab (AB154), Tiragolumab, Belres totug, 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.

[0133] 13. The FAK inhibitor, microtubule inhibitor and immune checkpoint inhibitor according to embodiment 1, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is docetaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, especially the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is docetaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0134] 14. The FAK inhibitor, microtubule inhibitor and immune checkpoint inhibitor according to embodiment 1, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is paclitaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, especially the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is paclitaxel; and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0135] 15. The FAK inhibitor, microtubule inhibitor, and immune checkpoint inhibitor of any one of embodiments 1-14, wherein the FAK inhibitor, the microtubule inhibitor, and the immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.

[0136] 16. A FAK inhibitor, a microtubule 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.

[0137] 17. A FAK inhibitor, a microtubule inhibitor, and an immune checkpoint inhibitor as described in any one of embodiments 1-16, wherein the tumor is breast cancer, ovarian cancer, or colon cancer (including colorectal cancer).

[0138] 18. A kit or pharmaceutically acceptable composition comprising:

[0139] (a) FAK inhibitors;

[0140] (b) microtubule inhibitors; and

[0141] (c) Immune checkpoint inhibitors.

[0142] 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:

[0143] 20. The kit or composition of embodiments 18-19, wherein the microtubule inhibitor is a taxane, Eribulin, Ixempra, or a vinca alkaloid.

[0144] 21. The kit or composition of any one of embodiments 18-20, wherein the microtubule inhibitor is a taxane.

[0145] 22. The kit or composition according to any one of embodiments 20-21, wherein the taxane is docetaxel, paclitaxel, cabazitaxel or cephalomannine, preferably docetaxel or paclitaxel.

[0146] 23. The kit or composition of any one of embodiments 18-22, wherein the microtubule inhibitor is docetaxel.

[0147] 24. The kit or composition of any one of embodiments 18-22, wherein the microtubule inhibitor is paclitaxel.

[0148] 25. The kit or composition of any one of embodiments 18-22, wherein the microtubule inhibitor is Eribulin.

[0149] 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 inhibitor.

[0150] 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, Camrelizumab, Sintilimab, Cemiplimab, envafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005, or MX-10181.

[0151] 28. The kit or composition according to any one of embodiments 18-26, 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.

[0152] 29. The kit or composition of any one of embodiments 18-26, wherein the immune checkpoint inhibitor is a TIGIT inhibitor, and further, the TIGIT inhibitor is Ociperlimab (BGB-A1217), Vibostolimab, domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036, or IBI-321.

[0153] 30. The kit or composition according to embodiment 18, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is docetaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, especially the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is docetaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0154] 31. The kit or composition according to embodiment 18, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is paclitaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, especially the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is paclitaxel; and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0155] 32. The kit or composition of any one of embodiments 18-31, wherein the composition is for use in medicine.

[0156] 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.

[0157] 34. The kit or composition of embodiment 33, wherein the tumor is breast cancer, ovarian cancer, or colon cancer (including colorectal cancer).

[0158] 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 microtubule inhibitor, and an immune checkpoint inhibitor.

[0159] 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:

[0160] 37. The method of embodiments 35-36, wherein the microtubule inhibitor is a taxane, Eribulin, Ixempra, or a vinca alkaloid.

[0161] 38. The method of any one of embodiments 35-37, wherein the microtubule inhibitor is a taxane.

[0162] 39. The method according to any one of embodiments 37-38, wherein the taxane is docetaxel, paclitaxel, cabazitaxel or cephalomannine, preferably docetaxel or paclitaxel.

[0163] 40. The method of any one of embodiments 35-39, wherein the microtubule inhibitor is docetaxel.

[0164] 41. The method of any one of embodiments 35-39, wherein the microtubule inhibitor is paclitaxel.

[0165] 42. The method of any one of embodiments 35-37, wherein the microtubule inhibitor is Eribulin.

[0166] 43. The method of 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 inhibitor.

[0167] 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, Camrelizumab, Sintilimab, Cemiplimab, envafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.

[0168] 45. The method according to any one of embodiments 35-43, 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.

[0169] 46. ​​The method according to any one of embodiments 35 to 43, wherein the immune checkpoint inhibitor is a TIGIT inhibitor, and further, the TIGIT inhibitor is Ociperlimab (BGB-A1217), Vibostolimab, domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.

[0170] 47. The method according to embodiment 35, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is docetaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, especially the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is docetaxel; and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0171] 48. The method according to embodiment 35, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is paclitaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, especially the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is paclitaxel; and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0172] 49. The method of any one of embodiments 35-48, wherein the FAK inhibitor, the microtubule inhibitor, and the immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.

[0173] 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.

[0174] 51. The method of any one of embodiments 35-50, wherein the tumor is breast cancer, ovarian cancer, or colon cancer (including colorectal cancer).

[0175] 52. A FAK inhibitor, a microtubule inhibitor, and an immune checkpoint inhibitor for use in a method of treating a tumor by increasing immunogenic cell death in a subject.

[0176] 53. The FAK inhibitor, microtubule 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:

[0177] 54. The FAK inhibitor, microtubule inhibitor and immune checkpoint inhibitor of embodiments 52-53, wherein the microtubule inhibitor is a taxane, Eribulin, Ixempra or a vinca alkaloid.

[0178] 55. The FAK inhibitor, microtubule inhibitor, and immune checkpoint inhibitor of any one of embodiments 52-54, wherein the microtubule inhibitor is a taxane.

[0179] 56. A FAK inhibitor, a microtubule inhibitor, and an immune checkpoint inhibitor as described in any one of embodiments 54-55, wherein the taxane is docetaxel, paclitaxel, cabazitaxel, or cephalomannine, preferably docetaxel or paclitaxel.

[0180] 57. The FAK inhibitor, microtubule inhibitor, and immune checkpoint inhibitor of any one of embodiments 52-56, wherein the microtubule inhibitor is docetaxel.

[0181] 58. A FAK inhibitor, a microtubule inhibitor, and an immune checkpoint inhibitor as described in any one of embodiments 52-56, wherein the microtubule inhibitor is paclitaxel.

[0182] 59. The FAK inhibitor, microtubule inhibitor, and immune checkpoint inhibitor of any one of embodiments 52-54, wherein the microtubule inhibitor is Eribulin.

[0183] 60. The FAK inhibitor, microtubule inhibitor, and immune checkpoint inhibitor of 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 inhibitor.

[0184] 61. The FAK inhibitor, microtubule inhibitor, and immune checkpoint inhibitor according to any one of embodiments 52 to 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, durvalumab, Avelumab, Camrelizumab, Sintilimab, Cemiplimab, envafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005, or MX-10181.

[0185] 62. The FAK inhibitor, microtubule inhibitor, and immune checkpoint inhibitor according to any one of embodiments 52-60, 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.

[0186] 63. The FAK inhibitor, microtubule inhibitor and immune checkpoint inhibitor according to any one of embodiments 52-60, wherein the immune checkpoint inhibitor is a TIGIT inhibitor, and further, the TIGIT inhibitor is 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.

[0187] 64. The FAK inhibitor, microtubule inhibitor and immune checkpoint inhibitor according to embodiment 52, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is docetaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, especially the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is docetaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0188] 65. The FAK inhibitor, microtubule inhibitor and immune checkpoint inhibitor according to embodiment 52, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is paclitaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, especially the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is paclitaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0189] 66. The FAK inhibitor, microtubule inhibitor, and immune checkpoint inhibitor of any one of embodiments 52-65, wherein the FAK inhibitor, the microtubule inhibitor, and the immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.

[0190] 67. A FAK inhibitor, a microtubule 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 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.

[0191] 68. A FAK inhibitor, a microtubule inhibitor, and an immune checkpoint inhibitor as described in any one of embodiments 52-67, wherein the tumor is breast cancer, ovarian cancer, or colon cancer (including colorectal cancer).

[0192] 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 microtubule inhibitor, and an immune checkpoint inhibitor.

[0193] 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, wherein the structure of IN10018 is as follows:

[0194] 71. The method of embodiments 69-70, wherein the microtubule inhibitor is a taxane, Eribulin, Ixempra, or a vinca alkaloid.

[0195] 72. The method of any one of embodiments 69-71, wherein the microtubule inhibitor is a taxane.

[0196] 73. The method according to any one of embodiments 71-72, wherein the taxane is docetaxel, paclitaxel, cabazitaxel or cephalomannine, preferably docetaxel or paclitaxel.

[0197] 74. The method of any one of embodiments 69-73, wherein the microtubule inhibitor is docetaxel.

[0198] 75. The method of any one of embodiments 69-73, wherein the microtubule inhibitor is paclitaxel.

[0199] 76. The method of any one of embodiments 69-73, wherein the microtubule inhibitor is Eribulin.

[0200] 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 inhibitor.

[0201] 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, Camrelizumab, Sintilimab, Cemiplimab, envafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.

[0202] 79. The 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.

[0203] 80. The method according to any one of embodiments 69 to 77, wherein the immune checkpoint inhibitor is a TIGIT inhibitor, and further, the TIGIT inhibitor is Ociperlimab (BGB-A1217), Vibostolimab, domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.

[0204] 81. The method according to embodiment 69, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is docetaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, a PD-1 / PD-L1 small molecule inhibitor, in particular, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is docetaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0205] 82. The method according to embodiment 69, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is paclitaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, in particular, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is paclitaxel; and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0206] 83. The method of any one of embodiments 69-82, wherein the FAK inhibitor, the microtubule inhibitor, and the immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.

[0207] 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.

[0208] 85. The method of any one of embodiments 69-84, wherein the tumor is breast cancer, ovarian cancer, or colon cancer (including colorectal cancer).

[0209] 86. Use of a FAK inhibitor in the preparation of a medicament for treating a tumor in a subject, wherein the FAK inhibitor, a microtubule inhibitor, and an immune checkpoint inhibitor are administered to the subject.

[0210] 87. Use of a microtubule inhibitor in the preparation of a medicament for treating a tumor in a subject, wherein a FAK inhibitor, the microtubule inhibitor and an immune checkpoint inhibitor are administered to the subject.

[0211] 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 microtubule inhibitor and the immune checkpoint inhibitor are administered to the subject.

[0212] 89. Use of FAK inhibitors, microtubule inhibitors and immune checkpoint inhibitors in the preparation of combined drugs for treating tumors.

[0213] 90. Use of a FAK inhibitor in the preparation of a drug for use in combination with a microtubule inhibitor and an immune checkpoint inhibitor in treating tumors.

[0214] 91. Use of a microtubule inhibitor in the preparation of a combination drug for treating tumors with a FAK inhibitor and an immune checkpoint inhibitor.

[0215] 92. Use of immune checkpoint inhibitors in the preparation of drugs for use in combination with FAK inhibitors and microtubule inhibitors in treating tumors.

[0216] 93. Use of FAK inhibitors, microtubule inhibitors and immune checkpoint inhibitors in the preparation of drugs for combined treatment of tumors.

[0217] 94. Use of a FAK inhibitor in the preparation of a medicament for treating tumors in combination with a microtubule inhibitor and an immune checkpoint inhibitor.

[0218] 95. Use of a microtubule inhibitor in the preparation of a drug for treating tumors in combination with a FAK inhibitor and an immune checkpoint inhibitor.

[0219] 96. Use of immune checkpoint inhibitors in the preparation of drugs for treating tumors in combination with FAK inhibitors and microtubule inhibitors.

[0220] 97. The use according to any one of embodiments 86 to 96, 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:

[0221] 98. The use according to any one of embodiments 86-97, wherein the microtubule inhibitor is a taxane, Eribulin, Ixempra or a vinca alkaloid.

[0222] 99. The use according to any one of embodiments 86-98, wherein the microtubule inhibitor is a taxane.

[0223] 100. The use according to any one of embodiments 98-99, wherein the taxane is docetaxel, paclitaxel, cabazitaxel or cephalomannine, preferably docetaxel or paclitaxel.

[0224] 101. The use according to any one of embodiments 98-100, wherein the microtubule inhibitor is docetaxel.

[0225] 102. The use according to any one of embodiments 98-100, wherein the microtubule inhibitor is paclitaxel.

[0226] 103. The use according to any one of embodiments 98-100, wherein the microtubule inhibitor is Eribulin.

[0227] 104. The use according to any one of embodiments 98-103, wherein the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, a PD-1 / PD-L1 small molecule inhibitor, or a TIGIT inhibitor.

[0228] 105. The use according to any one of embodiments 98 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, Camrelizumab, Sintilimab, Cemiplimab, envafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.

[0229] 106. The use according to any one of embodiments 98-104, 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.

[0230] 107. The use according to any one of embodiments 98 to 104, wherein the immune checkpoint inhibitor is a TIGIT inhibitor, and further, the TIGIT inhibitor is Ociperlimab (BGB-A1217), Vibostolimab, domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.

[0231] 108. The use according to any one of embodiments 98 to 107, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is docetaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, a PD-1 / PD-L1 small molecule inhibitor, in particular, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is docetaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0232] 109. The use according to any one of embodiments 98 to 107, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is paclitaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody, a PD-1 / PD-L1 small molecule inhibitor, especially the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is paclitaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

[0233] 110. The use according to any one of embodiments 98-109, wherein the FAK inhibitor, the microtubule inhibitor, and the immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.

[0234] 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.

[0235] 112. The use according to any one of embodiments 98 to 111, wherein the tumor is breast cancer, ovarian cancer or colon cancer (including colorectal cancer).

[0236] Example

[0237] 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.

[0238] The experimental methods in the following examples where specific conditions are not specified can be carried out according to conventional conditions of such reactions or according to the conditions recommended by the manufacturers. The experimental materials and reagents used in the following examples can be obtained from commercial channels unless otherwise specified.

[0239] The abbreviations used in the examples have the following meanings:

[0240] Example 1: Study on the Enhancement of Cell Death of Ovarian Cancer Cells Against Docetaxel Immunogenicity by FAK Target Inhibition

[0241] Experimental plan:

[0242] 1. FAK silencing combined with Docetaxel promotes apoptosis of ovarian cancer cells HCC827

[0243] In vitro experiments used siRNA technology to reduce the expression level of FAK and combined it with Docetaxel for 48 hours; flow cytometry was used to detect apoptosis in control (DMSO) and FAK silenced cells.

[0244] 2. FAK silencing combined with Docetaxel can effectively promote ICD in ovarian cancer cells SK-OV-3

[0245] In vitro experiments used siRNA technology to reduce the expression level of FAK, combined with Docetaxel for 48 hours, and flow cytometry was used to detect the expression of Calreticulin, the main target of ICD.

[0246] Experimental antibody: recombinant Alexa 647 fluorescent Anti-Calreticulin antibody (Abcam, ab196159), Annexin V apoptosis detection kit (Invitrogen, A35110).

[0247] FAK siRNA was provided by Genema, and the sequence is shown in the following table:

[0248] F: CCUGUAUGCCUAUCAGCUUTT;

[0249] R:AAGCUGAUAGGCAUACAGGTT

[0250] Experimental instruments:

[0251] Fluorescence microscope (Olympus U-HGLGPS) and chemiluminescence imager (BIORAD chemidoc touch)

[0252] Experimental results:

[0253] 1. FAK silencing combined with Docetaxel significantly increased SK-OV-3 cell apoptosis

[0254] SK-OV-3 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 0.3 μM or 1 μM Docetaxel for 48 hours. Cells were stained using an Annexin V kit and analyzed by flow cytometry. Apoptosis was calculated and compared with the control group and plotted using Graphpad 8.0. The results showed that Docetaxel treatment significantly enhanced apoptosis in the FAK silencing group compared to the control group, as shown in Figure 1.

[0255] 2. FAK silencing combined with Docetaxel significantly upregulated the release and exposure of the ICD target calreticulin

[0256] SK-OV-3 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 1 μM docetaxel for 48 hours. Fluorescence staining with a calreticulin antibody was performed and analyzed by flow cytometry. FlowJo software analysis revealed that the release and exposure of calreticulin were significantly enhanced after FAK silencing and the combination of docetaxel, as shown in Figure 2.

[0257] Example 2: Study on the Enhancement of Cell Death of Breast Cancer Cells Against Eribulin Immunogenicity by FAK Target Inhibition

[0258] Experimental plan:

[0259] 1. FAK silencing combined with Eribulin promotes apoptosis of breast cancer cells MDA-MB-231

[0260] In vitro experiments used siRNA technology to reduce the expression level of FAK and combined it with Eribulin for 48 hours; flow cytometry was used to detect apoptosis in control and FAK-silenced cells.

[0261] 2. FAK silencing combined with Eribulin can effectively promote ICD in breast cancer cells MDA-MB-231

[0262] In vitro experiments used siRNA technology to reduce the expression level of FAK and combined it with Eribulin for 48 hours; flow cytometry was used to detect the expression of Calreticulin, the main target of ICD.

[0263] Experimental Antibodies:

[0264] Restructuring Alexa 647 fluorescent Anti-Calreticulin antibody (Abcam, ab196159), Annexin V apoptosis detection kit (Invitrogen, A35110).

[0265] FAK siRNA was provided by Genema, and the sequence is shown in the following table:

[0266] F: CCUGUAUGCCUAUCAGCUUTT;

[0267] R:AAGCUGAUAGGCAUACAGGTT

[0268] Experimental instruments:

[0269] Fluorescence microscope (Olympus U-HGLGPS) and chemiluminescence imager (BIORAD chemidoc touch).

[0270] Experimental results:

[0271] 1. FAK silencing combined with Eribulin significantly increased apoptosis in MDA-MB-231 cells

[0272] 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 0.15 μM eribulin 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 eribulin treatment significantly enhanced both early and late apoptosis in the FAK silencing group compared with the control group, as shown in Figure 3.

[0273] 2. FAK silencing combined with Eribulin significantly upregulated the release and exposure of the ICD target calreticulin

[0274] 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 0.15 μM eribulin for 48 hours. Calreticulin staining was performed using a calreticulin antibody and analyzed by flow cytometry. FlowJo software analysis revealed that FAK silencing combined with eribulin significantly enhanced calreticulin release and exposure, as shown in Figure 4.

[0275] Example 3: Synergistic Effect of IN10018 and Docetaxel in Colon Cancer CT26 Cells

[0276] 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 Docetaxel was added. Ten drug concentrations were set up, starting with a 30 μM concentration, diluted threefold, and ending with a control with zero drug concentration. 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 incubated at 37°C in a 5% CO2 incubator for 72 hours.

[0277] 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 Docetaxel group was 0.19 μM; the IC50 for the Docetaxel + 5 μM IN10018 group was <0.005 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.

[0278] Example 4: Study of IN10018 and Docetaxel in Colon Cancer CT26 Cells

[0279] CT26 cells (Institute of Cell Biology, Chinese Academy of Sciences) were cultured in RPMI-1640 (Shanghai Yuanpei, catalog number: L210KJ, batch number: F210916) supplemented with 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, receiving culture medium; the second group received IN10018 at a concentration of 5 μM; the third group received Docetaxel (MCE, catalog number: HY-B0011, batch number: 111613) at a concentration of 0.1 μM; and the fourth group received a combination of IN10018 (5 μM) and Docetaxel (0.1 μM). The drugs were mixed and incubated at 37°C in a 5% CO2 incubator for 48 hours.

[0280] After 48 hours of drug treatment, cells were observed under a microscope and photographed, and the images 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 for 20 minutes at 4°C in the dark. After 20 minutes, cells were washed twice with flow cytometry buffer (PBS + 2% FBS). A cell apoptosis detection kit (Beyotime, catalog number: CL062L, batch number: 021921210811) was used, 195 μl of annexin V-FITC conjugate was added, the cells were gently mixed, 5 μl of annexin V-FITC antibody was added, and the mixture was gently mixed. 10 μl of PI dye was added, the mixture was incubated at room temperature in the dark for 15 minutes, and then analyzed on a flow cytometer.

[0281] Microscopic observation of cells revealed poor cell status in the Docetaxel monotherapy and dual-drug combination groups, with the dual-drug combination group showing the worst results, with more cell death. Cells in the control and IN10018 groups remained in good condition. The results showed that the CRT and Annexin V positivity rates in the dual-drug combination group were higher than those in the monotherapy groups (see Figures 6, 7a, and 7b for details).

[0282] Example 5: Study on the Induction of Immunogenic Cell Death Targets by Eribulin and IN10018 in Mouse Breast Cancer 4T1 Cells

[0283] The compound information is shown in Table 1.

[0284] Table 1:

[0285] The main reagent information of the experiment is shown in Table 2.

[0286] Table 2:

[0287] The experimental design is shown in Table 3.

[0288] Table 3:

[0289] Cell culture:

[0290] 4T1 cells were cultured as monolayers in RPMI-1640 medium supplemented with 10% fetal bovine serum at 37°C and 5% CO2. Cells were routinely digested and passaged using trypsin two to three times a week. When cells reached 80%-90% confluence and were in the exponential growth phase, they were harvested and plated.

[0291] 4T1 cells were trypsinized, harvested, and counted. Based on the count, the cells were diluted with RPMI-1640 + 10% FBS to a concentration of 50,000 cells / mL. The cells were then plated onto 12-well cell culture plates, with 2 mL of the cell suspension per well, or 100,000 cells. After plating, the cells were cultured in a 37°C, 5% CO2 incubator.

[0292] Add the test compound:

[0293] 24 hours after plating, test compounds IN10018 and Eribulin were added to different wells, respectively. The groups and drug concentrations are shown in Table 3.

[0294] Collect cells for flow cytometry:

[0295] After 48 hours of drug treatment, the cells were photographed using a microscope, and then trypsinized and collected for flow cytometry staining.

[0296] After washing the cells twice with flow cytometry buffer (DPBS + 2% FBS), each group of cells was divided into two equal portions. 0.5 μL of AF647 Anti-Calreticulin Antibody was added to each well of one portion, 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 detection. 0.5 μL of FITC was added to each well of the other portion. GRP94 Polyclonal Antibody was mixed and incubated at 4°C in the dark for 20 minutes. After that, the cells were washed twice with flow cytometry buffer and resuspended in 200 μL flow cytometry buffer before flow cytometry detection.

[0297] Data Analysis:

[0298] After the experiment, Flowjo (V10) software was used to analyze the cell positive rate.

[0299] Experimental results:

[0300] 48 hours after drug treatment, cells in each group were observed under a microscope. Eribulin monotherapy showed significant cell death, with the most significant cell death in the combination therapy group. Cell viability in the control and IN10018 groups was relatively good. Flow cytometric analysis showed that the positive rates of CRT, Annexin-V, and GRP94 in the combination therapy group were significantly higher than those in the monotherapy group. The relevant test results are shown in Figures 8a, 8b, and 8c.

[0301] Example 6: Study on the induction of immunogenic cell death targets by Eribulin and IN10018 in mouse ovarian epithelial cancer ID8 cells and human ovarian epithelial cancer TOV-21G cells in vitro

[0302] The information of compounds and main reagents is the same as that in Example 5.

[0303] The experimental design is shown in Table 4.

[0304] Table 4:

[0305] Cell culture:

[0306] ID8 and TOV-21G cells were cultured as monolayers in vitro. ID8 cells were cultured in DMEM medium supplemented with 10% fetal bovine serum, while TOV-21G cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum. Both were incubated 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.

[0307] ID8 cells and TOV-21G cells were digested with trypsin and then collected and counted. Based on the counting results, the cells were diluted with the corresponding complete culture medium to a concentration of 50,000 cells per milliliter. The cells were then plated on a 12-well cell culture plate, with 2 ml of cell suspension, or 100,000 cells, being plated per well.

[0308] After plating, the cells were cultured in a 37°C, 5% CO2 incubator.

[0309] Add the test compound:

[0310] 24 hours after plating, test compounds IN10018 and Eribulin were added to different wells, respectively. The groups and drug concentrations are shown in Table 4.

[0311] Collect cells for flow cytometry:

[0312] After 48 hours of drug treatment, the cells were photographed using a microscope, and then trypsinized and collected for flow cytometry staining.

[0313] 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.

[0314] Data Analysis:

[0315] After the experiment, Flowjo (V10) software was used to analyze the cell positive rate.

[0316] Experimental results:

[0317] Microscopic observation of the cells revealed significant cell death in both cell lines: the eribulin monotherapy group experienced significant cell death, with the combination group experiencing the most significant cell death. Cell viability in the control and IN10018 groups was relatively good. Flow cytometry analysis revealed significantly higher CRT and Annexin V positivity rates in the combination group than in the monotherapy group. Results for the ID8 cell line are shown in Figures 9a and 9b; those for the TOV-21G cell line are shown in Figures 10a and 10b.

[0318] Example 7: Study on the induction of immunogenic cell death targets by Docetaxel / Eribulin / Paclitaxel and Defactinib in mouse colon cancer CT26 cells in vitro

[0319] The compound information is shown in Table 5.

[0320] Table 5:

[0321] The main reagent information of the experiment is shown in Table 2:

[0322] The experimental design is shown in Table 6:

[0323] Table 6:

[0324] Cell culture:

[0325] CT26 cells were cultured as monolayers in RPMI-1640 medium supplemented with 10% fetal bovine serum at 37°C and 5% CO2. Cells were routinely digested and passaged using trypsin two to three times a week. When cells reached 80%-90% confluence and were in the exponential growth phase, they were harvested and plated.

[0326] CT26 cells were digested with trypsin and then collected and counted. Based on the counting results, the cells were diluted with the corresponding complete culture medium to a concentration of 50,000 cells per milliliter. The cells were then plated on a 12-well cell culture plate, with 2 ml of cell suspension (i.e., 100,000 cells) per well.

[0327] After plating, the cells were cultured in a 37°C, 5% CO2 incubator.

[0328] 24 hours after plating, the test compounds Docetaxel, Eribulin, Paclitaxel and Defactinib were added to different wells, respectively. The groups and drug concentrations are shown in Table 6.

[0329] After 48 hours of drug treatment, the cells were photographed using a microscope, and then trypsinized and collected for flow cytometry staining.

[0330] 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.

[0331] Data Analysis:

[0332] After the experiment, Flowjo (V10) software was used to analyze the cell positive rate.

[0333] Experimental results:

[0334] This study evaluated the effects of Docetaxel / Eribulin / Paclitaxel alone and in combination with Defactinib on inducing the expression of immune cell death targets in CT26 cells in vitro.

[0335] 48 hours after drug treatment, cells in each group were observed under a microscope. Cell death was evident in the docetaxel / eribulin / paclitaxel monotherapy group, with the most significant cell death in the combination therapy group. Cell viability in the control group was relatively good, while cell death was more pronounced in the defactinib monotherapy group. Flow cytometric analysis showed that the CRT and Annexin V positivity rates in the combination therapy group were significantly higher than those in the monotherapy group. The results for the docetaxel experiment are shown in Figures 11a and 11b; the results for the eribulin experiment are shown in Figures 12a and 12b; and the results for the paclitaxel experiment are shown in Figures 13a and 13b.

[0336] Example 8: Study on the Induction of Immunogenic Cell Death Targets by Docetaxel / Eribulin and Defactinib in Mouse Breast Cancer 4T1 Cells in Vitro

[0337] The compound information is shown in Table 5 (without Paclitaxel), and the main experimental reagent information is shown in Table 2.

[0338] The experimental design is shown in Table 7:

[0339] Table 7:

[0340] Cell culture:

[0341] 4T1 cells were cultured as monolayers in RPMI-1640 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 and were in the exponential growth phase, they were harvested and plated.

[0342] 4T1 cells were digested with trypsin and then collected and counted. Based on the counting results, the cells were diluted with the corresponding complete culture medium to a concentration of 50,000 cells per milliliter. The cells were then plated on a 12-well cell culture plate, with 2 ml of cell suspension (i.e., 100,000 cells) per well.

[0343] After plating, the cells were cultured in a 37°C, 5% CO2 incubator.

[0344] 24 hours after plating, the test compounds Defactinib, Docetaxel and Eribulin were added to different wells, respectively. The groups and drug concentrations are shown in Table 7.

[0345] Collect cells for flow cytometry:

[0346] After 48 hours of drug treatment, the cells were photographed using a microscope, and then trypsinized and collected for flow cytometry staining.

[0347] 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. Incubated at room temperature in the dark for 15 minutes before flow cytometry analysis.

[0348] Data Analysis:

[0349] After the experiment, Flowjo (V10) software was used to analyze the cell positive rate.

[0350] Experimental results:

[0351] 48 hours after drug treatment, cells in each group were observed under a microscope. Significant cell death was observed in the docetaxel / eribulin monotherapy group, with the most significant cell death in the combination therapy group. Cell viability in the control group was relatively good, while cell death was more pronounced in the defactinib monotherapy group. Flow cytometric analysis showed that both CRT and Annexin V positivity rates were significantly higher in the combination therapy group than in the Defactinib monotherapy group. The results of the docetaxel experiment are shown in Figures 14a and 14b; the results of the eribulin experiment are shown in Figures 15a and 15b.

[0352] Example 9: In vivo antitumor efficacy study of docetaxel injection in a BALB / c mouse subcutaneous transplant model of breast cancer 4T1 cells

[0353] Test materials:

[0354] Mice: Female BALB / c mice aged 6-8 weeks were purchased from Shanghai Slake Laboratory Animal Co., Ltd. Upon arrival, the animals were acclimated to the experimental environment before the experiment began. The animals were housed in an SPF-grade animal room using IVC (independent ventilation system) cages (5 per cage). All cages, bedding, and drinking water were sterilized before use. All experimental personnel were required to wear protective clothing and latex gloves when working in the animal room. Cages, feed, and drinking water were changed twice a week. The housing environment and lighting conditions were as follows:

[0355] Temperature: 20-26℃

[0356] Humidity: 40-70%

[0357] Photoperiod: 12 hours of light, 12 hours of no light

[0358] Cages: Made of polycarbonate, 300 mm × 180 mm × 150 mm. Bedding is corn cobs, changed twice a week.

[0359] Food: The experimental animals had free access to food (irradiated sterilized, dry pelleted food) throughout the experimental period.

[0360] Drinking water: Experimental animals can drink sterile water freely.

[0361] Cage identification: The animal information card for each cage should indicate the number of animals in the cage, sex, strain, receipt date, dosing regimen, experiment number, group and start date of the experiment.

[0362] Animal identification: Experimental animals were identified with ear tags.

[0363] Compound information is shown in Table 8

[0364] Table 8:

[0365] Breast cancer 4T1 cells (sourced from Nanjing Kebai Biotechnology Co., Ltd., catalog number CBP60352) were maintained and passaged by Ins Biotechnology (Nanjing) Co., Ltd. Cells were cultured as monolayers in RPMI-1640 medium supplemented with 10% fetal bovine serum at 37°C in a 5% CO2 incubator. Cells were routinely digested and passaged using trypsin-EDTA two to three times weekly. When cells were in the exponential growth phase and at 80%-90% saturation, they were harvested, counted, and plated.

[0366] Cell seeding and grouping

[0367] 0.1 mL contains 2×10 5 A cell suspension of 100 cells was subcutaneously inoculated on the right back of each mouse. 3 At 10 days after cell inoculation, the patients were randomly divided into groups and given medication according to the tumor volume. The grouping information is shown in Table 9.

[0368] Table 9. Dosage regimen of the test substance in 4T1 mouse transplant tumor model

[0369] Note: 1. Number of mice in each group;

[0370] 2. Dosing volume: 10 mL / kg based on mouse body weight. If the body weight decreases by more than 15%, stop dosing the animal; resume dosing when the body weight recovers to 10% of the decrease.

[0371] The preparation of the test substances is detailed in Table 10.

[0372] Table 10.

[0373] Daily observation of experimental animals

[0374] 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.

[0375] Experiment terminated

[0376] If the animal's health condition continues to deteriorate, or the tumor volume exceeds 3000mm 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 2000mm 3 The 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.

[0377] Tumor measurements and experimental parameters

[0378] Tumor diameter was measured with a vernier caliper three times a week. Tumor volume was calculated using the formula: V = 0.5 × a × b 2 , a and b represent the long diameter and short diameter of the tumor, respectively.

[0379] The tumor growth inhibition rate (TGI) (%) was calculated based on the tumor volume on the first day after grouping according to the following formula: TGI (%) = [1 - (average tumor volume of a given dosing group - average tumor volume of that dosing group at the start of treatment) / (average tumor volume of the vehicle control group - average tumor volume of the vehicle control group at the start of treatment)] × 100%.

[0380] Statistical analysis

[0381] 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.

[0382] Experimental results

[0383] The in vivo efficacy of the test substances DTX and / or MAX10181 in combination with IN10018 in a BALB / c mouse subcutaneous transplant tumor model of 4T1 mouse breast cancer cells. After cell inoculation, tumor growth was observed daily. Groups were divided according to tumor volume on the 10th day after inoculation. The average tumor volume in the group was approximately 52 mm. 3 Due to tumor burden, the control group was euthanized on day 31 after inoculation, which was 21 days after group administration, and the entire experiment was terminated.

[0384] On day 21 after group administration, the tumor volume of the control group was 2129.2±322.0mm 3 The tumor volumes of the DTX (10 mg / kg), MAX10181 (120 mg / kg), DTX+IN10018 (10+25 mg / kg), DTX+MAX10181 (10+120 mg / kg), and DTX+IN10018+MAX10181 (10+25+120 mg / kg) treatment groups were 1818.7±644.0 mm 3 , 2205.7±912.7mm 3 , 1182.9±264.3mm 3 , 1480.3±345.6mm 3 and 829.4±249.1mm 3 , see Table 11 for details. Compared with the control group, the comprehensive tumor volume showed that the tumor inhibition rates TGI of the DTX (10 mg / kg), MAX10181 (120 mg / kg), DTX+IN10018 (10+25 mg / kg), DTX+MAX10181 (10+120 mg / kg) and DTX+IN10018+MAX10181 (10+25+120 mg / kg) groups were 14.9% (p=0.0147), -3.7% (p=0.5642), 45.6% (p<0.0001), 31.3% (p<0.0001) and 62.6% (p<0.0001), respectively. See Table 11 for details. Statistical analysis was performed to compare the combined tumor volume with that of the DTX + IN10018 + MAX10181 (10 + 25 + 120 mg / kg) triple-drug combination group. The P values ​​for the control group, DTX (10 mg / kg), MAX10181 (120 mg / kg), DTX + IN10018 (10 + 25 mg / kg), and DTX + MAX10181 (10 + 120 mg / kg) groups were p < 0.0001, p < 0.0001, p < 0.0001, p = 0.0055, and p < 0.0001, respectively. The tumor volumes of each dose group at different time periods are shown in Figure 16.

[0385] Table 11: Data on day 21 after group administration

[0386] Note: 1. Calculated based on the number of days after group administration. Data are mean ± standard error.

[0387] 2. *: p < 0.05, ****: p < 0.0001, vs. control group, Two-way ANOVA.

[0388] 3. **: p < 0.01, ****: p < 0.0001, vs. DTX + IN10018 + MAX10181 (10 + 25 + 120 mg / kg) group, Two-way ANOVA.

[0389] The experiment was conducted according to the dosing schedule. Animals were observed daily for food and water intake, and their body weights were recorded three times a week. The body weight curves are shown in Figure 17. Throughout the dosing period, animals in the MAX10181-treated groups experienced diarrhea, which resulted in one death in each of the G3 MAX10181 (120 mg / kg) and G5 DTX + MAX10181 (10 + 120 mg / kg) groups. Additionally, weight loss was observed in the DTX + IN10018 + MAX10181 (10 + 25 + 120 mg / kg) groups, but it did not exceed 10%. Animals in all groups maintained good mental and motor function and tolerated all dosing routes.

[0390] in conclusion

[0391] Compared with the blank control group, the DTX (10 mg / kg), DTX+IN10018 (10+25 mg / kg), DTX+MAX10181 (10+120 mg / kg) and DTX+IN10018+MAX10181 (10+25+120 mg / kg) treatment groups all had significant tumor growth inhibition effects, and there were statistical differences compared with the control group. Comprehensively considering the entire dosing cycle, the tumor volume of the DTX+IN10018 (10+25 mg / kg) group was smaller than that of the DTX (10 mg / kg) single-drug group, indicating that the combination of DTX and IN10018 has better efficacy than single drugs; the tumor volume of the DTX+IN10018+MAX10181 (10+25+120 mg / kg) triple-drug combination group was statistically different from each single-drug group and each two-drug combination group, and compared with the other groups, indicating that the combination of DTX, IN10018 and MAX10181 has a better effect in inhibiting tumor growth. Although MAX10181 caused diarrhea in animals after administration, and led to the death of one animal in each of the MAX10181 (120 mg / kg) and DTX+MAX10181 (10+120 mg / kg) groups, the mental and motor conditions of the remaining animals were good, indicating that the animals showed a certain tolerance to the combination of DTX+IN10018+MAX10181 (10+25+120 mg / kg).

[0392] Example 10: Study on the induction of immunogenic cell death targets by docetaxel injection and AMP945 in mouse breast cancer 4T1 cells in vitro

[0393] Experimental Materials:

[0394] 1) Drugs used in this experiment

[0395] Docetaxel injection was provided by MCE, Lot No.: 111613

[0396] AMP945 provided by MCE, Lot No.:143253

[0397] 2) Antibodies used in this experiment

[0398] 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).

[0399] Experimental methods:

[0400] 4T1 cells (source: Nanjing Kebai Biotechnology Co., Ltd., catalog number: CBP60352) were cultured in RPMI-1640 (Shanghai Yuanpei, Cat No.: L210KJ, Lot No.: F210916) + 10% FBS (Gibco, Cat No.: 10099-141c, Lot No.: 2158737cp) at 37°C and 5% CO2. Trypsin was used for routine passage two to three times a week. 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 RPMI-1640 + 10% FBS to a concentration of 50,000 cells / mL and plated into 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 docetaxel injection (Docetaxel) at a concentration of 0.3 μM; the fifth group was treated with AMP945 (3 μM) combined with Docetaxel (0.3 μM); and the sixth group was treated with AMP945 (6 μM) combined with Docetaxel (0.3 μM). The drugs were mixed and cultured at 37°C in a 5% CO2 incubator for 48 hours.

[0401] Experimental results

[0402] After 48 hours of drug treatment, cells were collected for flow 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. Incubated at 4°C in the dark for 20 minutes, flow cytometry buffer was added, and 195 μl of Annexin-V-FITC binding solution was added using the Annexin staining kit (Beyotime). After pipetting and mixing with the cells, 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, and the samples were sent to the flow cytometer for signal measurement.

[0403] The results of flow cytometry analysis showed that the CRT positivity rate and Annexin-V positivity rate of the combination of the two drugs were significantly better than those of the single drug group and the control group, as shown in Figures 18a and 18b.

[0404] Example 11: Study on the induction of immunogenic cell death targets by Eribulin and AMP945 in mouse breast cancer 4T1 cells in vitro

[0405] Experimental Materials:

[0406] 1) Drugs used in this experiment

[0407] Eribulin was provided by Beijing Huaguan Pharmaceutical Technology Co., Ltd., Lot No.: HG-000000011-000-001

[0408] AMP945 provided by MCE, Lot No.:143253

[0409] 2) Antibodies used in this experiment

[0410] 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).

[0411] Experimental methods:

[0412] 4T1 cells (source: Nanjing Kebai Biotechnology Co., Ltd., catalog number: CBP60352) were cultured in RPMI-1640 (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 RPMI-1640 + 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 eribulin at a concentration of 0.3 μM; the fifth group received a combination of AMP945 (3 μM) and eribulin (0.3 μM); and the sixth group received a combination of AMP945 (6 μM) and eribulin (0.3 μM). The drugs were mixed and cultured at 37°C in a 5% CO2 incubator for 48 hours.

[0413] Experimental results

[0414] After 48 hours of drug treatment, cells were collected for flow 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. Incubated at 4°C in the dark for 20 minutes, flow cytometry buffer was added, and 195 μl of Annexin-V-FITC binding solution was added using the Annexin staining kit (Beyotime). After pipetting and mixing with the cells, 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, and the samples were sent to the flow cytometer for signal measurement.

[0415] The results of flow cytometry analysis showed that the CRT positivity rate and Annexin-V positivity rate of the combination of the two drugs were significantly better than those of the single drug group and the control group, as shown in Figures 19a and 19b.

[0416] 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 microtubule inhibitor, and an immune checkpoint inhibitor in the preparation of a medicament for treating a tumor in a subject.

2. A pharmaceutical combination product of a FAK inhibitor, a microtubule inhibitor, and an immune checkpoint inhibitor for use in treating a tumor in a subject.

3. A method for treating a tumor, comprising administering to a subject a therapeutically effective amount of a FAK inhibitor, a microtubule inhibitor, and an immune checkpoint inhibitor.

4. The use, pharmaceutical combination or method of any one of claims 1 to 3, wherein the FAK inhibitor and the microtubule inhibitor induce immunogenic cell death (ICD).

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 microtubule inhibitor is a taxane, eribulin, ixabepilone or a vinca alkaloid.

7. The use, pharmaceutical combination or method according to any one of claims 1 to 6, wherein the microtubule inhibitor is a taxane.

8. The use, pharmaceutical combination or method according to any one of claims 6 to 7, wherein the taxane is docetaxel, paclitaxel, cabazitaxel or cephalomannine, preferably docetaxel or paclitaxel.

9. The use, pharmaceutical combination or method according to any one of claims 1 to 8, wherein the microtubule inhibitor is docetaxel.

10. The use, pharmaceutical combination or method according to any one of claims 1 to 8, wherein the microtubule inhibitor is paclitaxel.

11. The use, pharmaceutical combination or method according to any one of claims 1 to 6, wherein the microtubule inhibitor is eribulin.

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 inhibitor.

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

16. The use, pharmaceutical combination product or method according to any one of claims 1 to 4, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is docetaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, in particular, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is docetaxel; and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

17. The use, pharmaceutical combination product or method according to any one of claims 1 to 4, wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is paclitaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, in particular, the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is paclitaxel; and the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

18. The use, pharmaceutical combination or method according to any one of claims 1 to 17, wherein the FAK inhibitor, the microtubule 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, ovarian cancer or colon cancer (including colorectal cancer).

21. A kit or pharmaceutically acceptable composition comprising: (a) FAK inhibitors; (b) microtubule inhibitors; and (c) Immune checkpoint inhibitors.

22. The kit or composition of claim 21, wherein the FAK inhibitor and the microtubule 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 according to any one of claims 21 to 23, wherein the microtubule inhibitor is a taxane, eribulin, ixabepilone, or a vinca alkaloid.

25. The kit or composition according to any one of claims 21 to 24, wherein the microtubule inhibitor is a taxane.

26. The kit or composition according to any one of claims 24-25, wherein the taxane is docetaxel, paclitaxel, cabazitaxel or cephalomannine, preferably docetaxel or paclitaxel.

27. The kit or composition of any one of claims 21 to 26, wherein the microtubule inhibitor is docetaxel.

28. The kit or composition of any one of claims 21-26, wherein the microtubule inhibitor is paclitaxel.

29. The kit or composition of any one of claims 21-24, wherein the microtubule inhibitor is Eribulin.

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 inhibitor.

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

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 microtubule inhibitor is docetaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, especially the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is docetaxel; the immune checkpoint inhibitor is 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 microtubule inhibitor is paclitaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody or a PD-1 / PD-L1 small molecule inhibitor, especially the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, the microtubule inhibitor is paclitaxel; the immune checkpoint inhibitor is an anti-PD-1 / PD-L1 antibody.

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

37. The 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, ovarian cancer, or colon cancer (including colorectal cancer).

39. FAK inhibitors for use in the treatment of tumors to enhance immunogenic cell death induced by microtubule inhibitors.

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 according to claim 39 or 40, wherein the microtubule inhibitor is a taxane, Eribulin, Ixempra or a vinca alkaloid.

42. The FAK inhibitor according to claim 41, wherein the taxane is docetaxel, paclitaxel, cabazitaxel or cephalomannine, preferably docetaxel or paclitaxel.

43. The FAK inhibitor of any one of claims 39-42, 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, metastasis caused by spindle cell carcinoma, anaplastic large cell lymphoma, anaplastic thyroid cancer, non-Hodgkin lymphoma, Hodgkin lymphoma, glioma, or hematological malignancy, for example 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; more preferably, the tumor is breast cancer, ovarian cancer or colon cancer (including colorectal cancer).

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