WO2024056074A1 - Fak抑制剂及拓扑异构酶抑制剂的药物组合及用途 - Google Patents

Fak抑制剂及拓扑异构酶抑制剂的药物组合及用途 Download PDF

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WO2024056074A1
WO2024056074A1 PCT/CN2023/119106 CN2023119106W WO2024056074A1 WO 2024056074 A1 WO2024056074 A1 WO 2024056074A1 CN 2023119106 W CN2023119106 W CN 2023119106W WO 2024056074 A1 WO2024056074 A1 WO 2024056074A1
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inhibitor
cancer
topoisomerase
pharmaceutically acceptable
fak
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English (en)
French (fr)
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张宝袁
刘学彬
高家明
张平
庞然
王在琪
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应世生物科技(南京)有限公司
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Priority to CN202380064083.8A priority Critical patent/CN119855608A/zh
Publication of WO2024056074A1 publication Critical patent/WO2024056074A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the invention belongs to the field of medicinal chemistry. Specifically, the present invention relates to focal adhesion kinase (Focal Adhesion Kinase, FAK) inhibitors combined with other drugs to treat tumors.
  • focal adhesion kinase Fecal Adhesion Kinase, FAK
  • Immunogenic cell death is based on the additive effect of programmed cell death.
  • This type of stress signal includes endoplasmic reticulum stress (ER stress) and reactive oxygen species stress (oxidative stress).
  • ER stress endoplasmic reticulum stress
  • oxidative stress reactive oxygen species stress
  • cells will first try to repair the stress. If the damage caused by the stress exceeds the ability to repair, the cells will initiate a programmed death process. This process is often accompanied by the release of a type of damage-associated molecules (Damage associated molecular patterns, DAMPs).
  • DAMPs damage associated molecular patterns
  • This type of DAMPs will be specifically recognized by pattern recognition receptors on Antigen-presenting cells (APCs) in the body, induce the maturation, differentiation and activation of APCs, and gradually present them to immune cells such as effector T cells, thereby It causes immune cells to generate antigen memory, and when tumor cells from the same source are found again, the immune cells will specifically recognize and kill the tumor cells.
  • the new tumor-specific immune response initiated by ICD can increase sensitivity to immune checkpoint inhibitors (Immuno-checkpoint inhibitors, ICI), thus enhancing the effect of immune checkpoint inhibitors and producing immune Memory for durable antitumor responses.
  • FAK also known as protein tyrosine kinase 2 (PTK2), is a non-receptor tyrosine kinase and a key component of the focal adhesion complex. FAK plays an important role in mediating integrin and growth factor signaling to regulate tumor cell invasion, proliferation, and survival.
  • PTK2 protein tyrosine kinase 2
  • topoisomerase inhibitors bind to the easily dissociable complex of the corresponding isomerase and form a stable complex, preventing the smooth passage of replication forks and causing DNA chain breakage, interfering with DNA replication and transcription. This type of drug is widely used in the treatment of tumors. However, these drugs will develop some resistance after being taken for a period of time.
  • Chemotherapy including a combination of topoisomerase inhibitors and immunotherapy, has been widely used in clinical practice. However, there is still room for improvement in efficacy, especially how to enhance the efficacy and produce lasting anti-tumor effects.
  • One aspect of the present disclosure provides the use of a FAK inhibitor, a topoisomerase inhibitor and an immune checkpoint inhibitor in the preparation of a medicament for treating a tumor in a subject, wherein the topoisomerase inhibitor is not an anthracycline kind.
  • Yet another aspect of the present disclosure provides a pharmaceutical combination product of a FAK inhibitor, a topoisomerase inhibitor and an immune checkpoint inhibitor for use in treating a tumor in a subject, wherein the topoisomerase inhibitor is not an anthracycline kind.
  • Yet another aspect of the present disclosure provides a method of treating tumors, the method comprising administering to a subject in need thereof a therapeutically effective amount of a FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor, wherein the topoisomerase
  • the enzyme inhibitor is not an anthracycline.
  • kit or pharmaceutically acceptable composition which includes: (a) a FAK inhibitor; (b) a topoisomerase inhibitor, the topoisomerase inhibitor is not anthracyclines; and (c) immune checkpoint inhibitors.
  • Yet another aspect of the present disclosure provides the use of a FAK inhibitor and a topoisomerase inhibitor in the preparation of a medicament for treating tumors, wherein the FAK inhibitor is used to enhance immunity induced by the topoisomerase inhibitor. Primary cell death, wherein the topoisomerase inhibitor is not an anthracycline.
  • Yet another aspect of the present disclosure provides FAK inhibitors for use in the treatment of tumors to enhance immunogenic cell death induced by a topoisomerase inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.
  • Yet another aspect of the present disclosure provides a method of treating tumors, the method comprising administering to a subject in need thereof a therapeutically effective amount of a FAK inhibitor and a topoisomerase inhibitor, wherein the FAK inhibitor is used to enhance the topoisomerase inhibitor.
  • a FAK inhibitor is used to enhance the topoisomerase inhibitor.
  • Yet another aspect of the present disclosure provides the use of a FAK inhibitor, a topoisomerase inhibitor and an immune checkpoint inhibitor in the preparation of a drug for combined treatment of tumors, wherein the topoisomerase inhibitor is not an anthracycline .
  • Yet another aspect of the present disclosure provides the use of a FAK inhibitor in the preparation of a medicament for the treatment of tumors in combination with a topoisomerase inhibitor and an immune checkpoint inhibitor, wherein the topoisomerase inhibitor is not an anthracycline kind.
  • Yet another aspect of the disclosure provides the use of a topoisomerase inhibitor in the preparation of a medicament for use in combination with a FAK inhibitor and an immune checkpoint inhibitor to treat tumors, wherein the topoisomerase inhibitor is not an anthracycline kind.
  • Yet another aspect of the present disclosure provides the use of an immune checkpoint inhibitor in the preparation of a medicament for the treatment of tumors in combination with a FAK inhibitor and a topoisomerase inhibitor, wherein the topoisomerase inhibitor is not an anthracycline kind.
  • Yet another aspect of the present disclosure provides the use of a FAK inhibitor in the preparation of a medicament for the treatment of tumors in combination with a topoisomerase inhibitor and an immune checkpoint inhibitor, wherein the topoisomerase inhibitor is not an anthracycline .
  • Yet another aspect of the present disclosure provides the use of a topoisomerase inhibitor in the preparation of a medicament for the treatment of tumors in combination with a FAK inhibitor and an immune checkpoint inhibitor, wherein the topoisomerase inhibitor is not an anthracycline .
  • Yet another aspect of the disclosure provides the use of an immune checkpoint inhibitor in the preparation of a medicament for the treatment of tumors in combination with a FAK inhibitor and a topoisomerase inhibitor, wherein the topoisomerase inhibitor is not an anthracycline .
  • kits which includes: a FAK inhibitor; and instructions, which indicate that the FAK inhibitor can be used to treat tumors in combination with a topoisomerase inhibitor and an immune checkpoint inhibitor, wherein the The above topoisomerase inhibitors are not anthracyclines.
  • kits which includes: a topoisomerase inhibitor; and instructions indicating that the topoisomerase inhibitor can be used to treat tumors in combination with FAK inhibitors and immune checkpoint inhibitors. , wherein the topoisomerase inhibitor is not an anthracycline.
  • kits which includes: an immune checkpoint inhibitor; and instructions indicating that the immune checkpoint inhibitor can be used to treat tumors in combination with a FAK inhibitor and a topoisomerase inhibitor, Wherein the topoisomerase inhibitor is not an anthracycline.
  • Another aspect of the present disclosure provides a method of treating a tumor, the method comprising administering to a subject in need thereof a therapeutically effective amount of a FAK inhibitor and a topoisomerase inhibitor, wherein the topoisomerase inhibitor is not anthracene Ring type.
  • Yet another aspect of the present disclosure provides a pharmaceutical combination product of a FAK inhibitor and a topoisomerase inhibitor for use in treating tumors in a subject in need thereof, wherein the topoisomerase inhibitor is not an anthracycline.
  • Yet another aspect of the present disclosure provides the use of a FAK inhibitor and a topoisomerase inhibitor in the preparation of a combination drug for treating tumors, wherein the topoisomerase inhibitor is not an anthracycline.
  • Yet another aspect of the present disclosure provides the use of a FAK inhibitor in the preparation of a medicament for use in combination with a topoisomerase inhibitor to treat tumors, wherein the topoisomerase inhibitor is not an anthracycline.
  • Yet another aspect of the present disclosure provides the use of a topoisomerase inhibitor in the preparation of a medicament for use in combination with a FAK inhibitor to treat tumors, wherein the topoisomerase inhibitor is not an anthracycline.
  • Yet another aspect of the present disclosure provides the use of a FAK inhibitor and a topoisomerase inhibitor in the preparation of a medicament for combined treatment of tumors, wherein the topoisomerase inhibitor is not an anthracycline.
  • Yet another aspect of the present disclosure provides the use of a FAK inhibitor in the preparation of a medicament for the treatment of tumors in combination with a topoisomerase inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.
  • Yet another aspect of the present disclosure provides the use of a topoisomerase inhibitor in the preparation of a medicament for the treatment of tumors in combination with a FAK inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.
  • kits which includes: a FAK inhibitor; and instructions indicating that the FAK inhibitor can be used to treat tumors in combination with a topoisomerase inhibitor, wherein the topoisomerase inhibitor
  • the agent is not an anthracycline.
  • kits which includes: a topoisomerase inhibitor; and instructions indicating that the topoisomerase inhibitor can be used to treat tumors in combination with a FAK inhibitor, wherein the topoisomerase inhibitor
  • the structural enzyme inhibitor is not an anthracycline.
  • 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.
  • the above acceptable salt is further preferably IN10018 or a pharmaceutically acceptable salt thereof, especially IN10018 tartrate.
  • the structure of IN10018 is as follows:
  • the Defactinib is also called defatinib, and its CAS number is 1345713-71-4; the CAS number of GSK2256098 is 1224887-10-8.
  • the CAS number of PF-00562271 is 717907-75-0; the CAS number of VS-4718 is 1061353-68-1; the APG-2449 is developed by Ascentage Pharmaceuticals; the CAS number of AMP945 is 1393653- 34-3.
  • the topoisomerase inhibitor is a topoisomerase I inhibitor.
  • the topoisomerase I inhibitor is Topotecan, Irinotecan, Belotecan, Cositecan, Exatecan (DX) -8951), Indenoisoquinoline (Indenoisoquinoline), Indotecan (LMP-400), Indimitecan (LMP-776), Simmitecan (Simmitecan), Gimatecan (ST1481), EC-112002 , PLX-038 (NK012) , AR-67, or a pharmaceutically acceptable salt thereof.
  • the CAS number of Topotecan is 123948-87-8 ; the CAS number of Irinotecan is 97682-44-5 ; the CAS number of Belotecan is 256411 -32-2 ; the CAS number of Cositecan is 401905-67-7 ; the CAS number of Exatecan (DX-8951) is 171335-80-1; The CAS number of Indenoisoquinoline is 97501-75-2; the CAS number of Indotecan (LMP-400) is 915303-09-2; the CAS number of Indimitecan (LMP-776) is 915360- 05-3; The CAS number of Simmitecan is 951290-31-6; the CAS number of Gimatecan (ST1481) is 292618-32-7; the EC-112002 is developed by Elucida Oncology Inc.
  • the CAS number of PLX-038 (NK012) is 86639-52-3; the CAS number of AR-67 is 220913-32-6.
  • the topoisomerase I inhibitor is irinotecan or a pharmaceutically acceptable salt thereof.
  • the topoisomerase inhibitor is a topoisomerase II inhibitor.
  • the topoisomerase II inhibitor is Etoposide, Teniposide, Sabarubicin, or a pharmaceutically acceptable salt thereof.
  • the CAS number of Etoposide is 33419-42-0 ; the CAS number of Teniposide is 29767-20-2 ; the CAS number of Sabarubicin is 211100-13-9 .
  • the topoisomerase II inhibitor is Etoposide, Teniposide, or a pharmaceutically acceptable salt thereof.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody, a PD-1/PD-L1 small molecule inhibitor or a TIGIT antibody.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody.
  • the anti-PD-1/PD-L1 antibody is pembrolizumab or tislelizumab. Tislelizumab, Nivolumab, Toripalimab, Atezolizumab, durvalumab, Avelumab, Avelumab Atezolizumab, Camrelizumab, Sintilimab, Cemiplimab, envafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.
  • the immune checkpoint inhibitor is a small molecule inhibitor of PD-1/PD-L1.
  • the small molecule inhibitor of PD-1/PD-L1 is INCB-086550, lazertinib ( Lazertinib), IMMH-010, CA-170, ABSK043 or RRx-001.
  • the immune checkpoint inhibitor is a TIGIT antibody.
  • the TIGIT antibody is Ociperlimab (Ociperlimab/BGB-A1217), Vibostolimab (Vibostolimab), domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939, COM -902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, and the topoisomerase inhibitor is irinotecan or a pharmaceutically acceptable salt thereof; the immune checkpoint inhibition
  • the agent is an anti-PD-1/PD-L1 antibody or a PD-1/PD-L1 small molecule inhibitor.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, and the topoisomerase inhibitor is irinotecan or a pharmaceutically acceptable salt thereof; the immune checkpoint inhibition
  • the agent is an anti-PD-1/PD-L1 antibody.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, and the topoisomerase inhibitor is Etoposide, Teniposide, or a pharmaceutically acceptable salt thereof. salt;
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or a small molecule inhibitor of PD-1/PD-L1.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, and the topoisomerase inhibitor is Etoposide, Teniposide, or a pharmaceutically acceptable salt thereof. salt; the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody.
  • the FAK inhibitor, the topoisomerase inhibitor and the immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.
  • the tumor is bladder cancer, breast cancer, cervical cancer, colon cancer (including colorectal cancer), esophageal cancer, esophageal squamous cell carcinoma, head and neck cancer, liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer).
  • melanoma myeloma, rhabdomyosarcoma, inflammatory myofibroblastic tumor, neuroblastoma, pancreatic cancer, prostate cancer, kidney cancer, renal cell carcinoma, sarcoma (including osteosarcoma), skin cancer (including squamous cell carcinoma) (like cell carcinoma), gastric cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal cancer, neuroendocrine cancer, ovarian cancer, salivary gland cancer, spindle cell carcinoma anaplastic large cell lymphoma, anaplastic thyroid carcinoma, non-Hodgkin lymphoma, Hodgkin lymphoma, glioma, or hematological malignancies such as acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), diffuse large B-cell lymphoma (AML), acute
  • the tumor is breast cancer or colon cancer (including colorectal cancer).
  • Figure 1 shows that combined treatment with 2 ⁇ M etoposide for 48 hours after FAK silencing increased MDA-MB-231 cell apoptosis.
  • Figure 2 shows that FAK silencing combined with 2 ⁇ M etoposide treatment for 48 hours enhanced calreticulin release and exposure.
  • Figure 3 shows that combined treatment with 2 ⁇ M teniposide for 48 hours after FAK silencing increased MDA-MB-231 cell apoptosis.
  • Figure 4 shows that FAK silencing combined with 2 ⁇ M teniposide treatment for 48 hours enhanced calreticulin release and exposure.
  • Figure 5 shows the IC50 value of etoposide combined with IN10018 in colon cancer cells CT26.
  • Figure 6 shows a white light microscope photo of colon cancer CT26 cells taken after incubation with drugs for 48 hours.
  • Figure 7 shows the situation of colon cancer CT26 cells after incubation with IN10018 and etoposide for 48 hours, in which: 7a shows the percentage of CRT-positive cells after incubation of colon cancer CT26 cells with drugs for 48 hours; 7b shows colon cancer CT26 cells with drugs Percentage of Annexin V-positive cells after 48 hours of incubation.
  • Figure 8 shows the situation after colon cancer CT26 cells were incubated with IN10018 and irinotecan for 48 hours, where: 8a shows the percentage of CRT-positive cells after colon cancer CT26 cells were incubated with drugs for 48 hours; 8b shows colon cancer CT26 cells with drugs Percentage of Annexin V-positive cells after 48 hours of incubation.
  • Figure 9 shows the curve of the inhibitory effect of teniposide and its combination with IN10018 on the proliferation of colon cancer CT26 cells relative to drug concentration.
  • Figure 10 shows the situation of colon cancer CT26 cells after incubation with IN10018 and teniposide for 48 hours, in which: 10a shows the percentage of Calreticulin (CRT) positive cells after mouse colon cancer CT26 cells were incubated with the drug for 48 hours; 10b shows the percentage of Calreticulin (CRT) positive cells. Percentage of Annexin-V positive cells after mouse colon cancer CT26 cells were incubated with drugs for 48 hours.
  • CRT Calreticulin
  • Figure 11 shows the tumor growth curve after administration of MC38 mouse colorectal cancer cell C57BL/6 mouse subcutaneous homograft tumor model, in which the data points represent the average tumor volume within the group, and the error bars represent the standard error (SEM).
  • Figure 12 shows the body weight change curve after administration of MC38 mouse colorectal cancer cell C57BL/6 mouse subcutaneous allograft tumor model, in which the data points represent the average body weight within the group, and the error bars represent the standard error (SEM).
  • Figure 13 shows the situation of mouse breast cancer T41 cells after incubation with AMP945 and Etoposide for 48 hours, where: 13a shows the percentage of Calreticulin (CRT) positive cells after mouse breast cancer T41 cells were incubated with drugs for 48 hours; 13b shows the mouse Percentage of Annexin-V positive cells after breast cancer T41 cells were incubated with drugs for 48 hours.
  • CRT Calreticulin
  • Figure 14 shows the situation of mouse breast cancer T41 cells after incubation with AMP945 and Irinotecan for 48 hours, where: 14a shows the percentage of Calreticulin (CRT) positive cells after mouse breast cancer T41 cells were incubated with drugs for 48 hours; 14b shows the mouse Percentage of Annexin-V positive cells after breast cancer T41 cells were incubated with drugs for 48 hours.
  • CRT Calreticulin
  • FAK inhibitor refers to a potent inhibitor of FAK, which may be suitable for mammals, especially humans.
  • the FAK inhibitor is IN10018, Defactinib, GSK2256098, PF-00562271, VS-4718, AMP945, AMP886 or a pharmaceutically acceptable salt thereof, and the structure of IN10018 is as follows:
  • the Defactinib is also called defatinib, and its CAS number is 1345713-71-4; the CAS number of GSK2256098 is 1224887-10-8.
  • the CAS number of PF-00562271 is 717907-75-0; the CAS number of VS-4718 is 1061353-68-1; the APG-2449 is developed by Ascentage Pharmaceuticals; the CAS number of AMP945 is 1393653- 34-3.
  • the FAK inhibitor is preferably IN10018, Defactinib, AMP945 or a pharmaceutically acceptable salt thereof.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof, Especially IN10018 tartrate.
  • topoisomerase inhibitor refers to a class of compounds capable of inhibiting the activity of DNA topoisomerase. Common ones include topoisomerase I inhibitors and topoisomerase II inhibitors.
  • anthracycline refers to a class of topoisomerase II inhibitors, including doxorubicin, epirubicin, daunorubicin, and aclarithromycin.
  • immune checkpoint inhibitor refers to drugs that can improve the activity of the immune system by regulating immune checkpoint pathways (such as PD-1, TIGIT, CTLA-4, LAG-3, TIM-3, etc.).
  • the immune checkpoint inhibitor is an antagonist of the PD-1/PD-L1 (programmed cell death protein 1) pathway (also known as a "PD-1 inhibitor”) or a TIGIT inhibitor.
  • PD-1 inhibitors are also referred to as PD-1/PD-L1 inhibitors in this disclosure.
  • the PD-1/PD-L1 inhibitor is a PD-1/PD-L1 antibody, including but not limited to pembrolizumab (Keytruda/ K drug), Tislelizumab (Tislelizumab/Baizian), Nivolumab (Nivolumab), Toripalimab (Toripalimab/Tuoyi), Atezolizumab (Taishengqi) ), durvalumab (Infinifer/durvalumab/durvalumab), avelumab (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.
  • pembrolizumab Keytruda/ K drug
  • Tislelizumab Tislelizuma
  • the PD-1 inhibitor is toripalimab. In some embodiments, PD-1 inhibitors are used to treat human subjects. In some embodiments, PD-1 is human PD-1.
  • PD-1/PD-L1 inhibitors also include PD-1/PD-L1 small molecule inhibitors, such as INCB-086550, Lazertinib, IMMH-010, CA-170, ABSK043 or RRx-001.
  • TIGIT also known as WUCAM, Vstm3, VSIG9 is a receptor of the Ig superfamily and a new immune checkpoint after PD-1/PD-L1.
  • the immune checkpoint inhibitor is a TIGIT inhibitor, including but not limited to Ociperlimab (Ociperlimab/BGB-A1217), Vibostolimab (Vibostolimab) ), domvanalimab(AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M- 6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.
  • TIGIT inhibitors are used to treat human subjects. To avoid ambiguity, antibodies in this article include double antibodies.
  • pharmaceutical combination or “pharmaceutical combination product” may refer to a fixed combination in the form of a dosage unit (for example, all active pharmaceutical ingredients are present in one dosage form) or a kit of products for combined administration. Refers to the combination of a drug and instructions indicating that the drug can be used in combination with one or more other drugs.
  • combination therapy or “combination drug” refers to the combination of one drug with one or more other drugs to treat a disease, including the combination of one drug with another one or more drugs. This also includes the combination of a drug and instructions indicating that the drug can be used in combination with one or more other drugs.
  • “Simultaneous or sequential administration” in this application refers to two or more drugs administered simultaneously or at a certain time interval within one administration cycle (for example, within 4 weeks, within 3 weeks, within 2 weeks, within 1 week, or within 24 hours). Administered sequentially, the methods of drug administration (such as oral, intravenous, intramuscular or subcutaneous administration, etc.) may be the same or different, and the administration frequency/period of two or more drugs may be the same or different. When the treatment methods, products or uses of the present disclosure involve two drugs, the two drugs can be administered separately at the same time or at certain intervals.
  • the three drugs can be administered at the same time point, or two drugs can be administered at one time point and one drug can be administered at another time point, or all Each of the three drugs was administered at different time points.
  • the PD-1/PD-L1 inhibitor is administered intravenously (eg, as an intravenous infusion) or subcutaneously, or orally.
  • the PD-1/PD-L1 inhibitor is administered as an intravenous infusion.
  • a TIGIT inhibitor is administered intravenously (eg, as an intravenous infusion) or subcutaneously, or orally.
  • the TIGIT inhibitor is administered as an intravenous infusion.
  • immune checkpoint inhibitors to treat cancer depends on the presence of tumor antigen-specific T cells within tumor tissue. This requires that tumor tissues express antigens that distinguish themselves from their non-transformed counterparts, for example, through novel protein products called neoantigens. Tumor neoantigen burden is closely related to immunogenicity and sensitivity (e.g., to checkpoint inhibitor therapies), meaning that less immunogenic tumors should be largely resistant to these agents. Therapies designed to release tumor antigens that can be taken up by APCs, such as those that induce immunogenic cell death (ICD), may promote effective anti-tumor immunity, especially when further combined with checkpoint inhibitors.
  • ICD immunogenic cell death
  • treatment refers to the administration of one or more pharmaceutical substances to a subject suffering from a disease or having symptoms of said disease in order to cure, alleviate, alleviate, alter, treat, ameliorate, ameliorate or affect said disease.
  • Disease or symptoms of said disease In some embodiments, the disease is tumor or cancer.
  • tumor refers to abnormal lesions formed by the body's abnormal growth caused by various tumorigenic factors. Cells in local tissues lose normal control of their growth at the genetic level, resulting in abnormal proliferation of their clonal types.
  • the tumors include, but are not limited to: bladder cancer, breast cancer, cervical cancer, colon cancer (including colorectal cancer), esophageal cancer, esophageal squamous cell carcinoma, head and neck cancer, liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer).
  • small cell lung cancer small cell lung cancer
  • melanoma myeloma, rhabdomyosarcoma, inflammatory myofibroblastic tumor, neuroblastoma, pancreatic cancer, prostate cancer, kidney cancer, renal cell carcinoma, sarcoma (including osteosarcoma), skin cancer (including Squamous cell carcinoma), gastric cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal cancer, neuroendocrine cancer, ovarian cancer, salivary gland cancer, spindle cell carcinoma Metastases, anaplastic large cell lymphoma, anaplastic thyroid carcinoma, non-Hodgkin lymphoma, Hodgkin lymphoma, glioma, or hematological malignancies such as acute myeloid leukemia (AML), acute lymphoblastic leukemia Leukemia (ALL), diffuse large B
  • the term "subject” or “subject” refers to mammals and non-mammals.
  • Mammal means any member of the class Mammalia, which includes, but is not limited to: humans; non-human primates, such as chimpanzees and other ape and monkey species; farm animals, such as cattle, horses, sheep, goats, and pigs; Domestic animals, such as rabbits, dogs, and cats; laboratory animals, including rodents, such as rats, mice, and guinea pigs; etc. Examples of non-mammals include, but are not limited to, birds.
  • the term "subject” does not identify a specific age or gender. In some embodiments, the subject is a human.
  • pharmaceutically acceptable means nontoxic, biologically tolerable, and suitable for administration to a subject.
  • pharmaceutically acceptable salts refers to nontoxic, biologically tolerable acid addition salts suitable for administration to a subject, including, but not limited to, acid addition salts with inorganic acids. , such as hydrochlorides, hydrobromides, carbonates, bicarbonates, phosphates, sulfates, sulfites, nitrates, etc.; and acid addition salts formed with organic acids, such as formates, acetates, etc.
  • Acid malate, maleate, fumarate, tartrate, succinate, citrate, lactate, methanesulfonate, p-toluenesulfonate, 2-hydroxyethanesulfonate , benzoate, salicylate, stearate and salts formed with alkanedicarboxylic acids of the formula HOOC-(CH 2 ) n -COOH (where n is 0-4), etc.
  • pharmaceutically acceptable acid addition salts can be obtained by dissolving the free base in a suitable solvent and treating the solution with an acid according to conventional procedures for preparing acid addition salts from basic compounds.
  • suitable solvent for preparing acid addition salts from basic compounds.
  • One skilled in the art will be able to identify, without undue experimentation, various synthetic methods that can be used to prepare nontoxic pharmaceutically acceptable acid addition salts.
  • composition means that it must be chemically and/or toxicologically compatible with the other ingredients including the formulation, and/or with the subject to be treated therewith.
  • therapeutically effective amount refers to an amount generally sufficient to produce a beneficial therapeutic effect in a subject. Conventional influencing factors (e.g., mode of administration, pharmacokinetics of the compound, severity and course of disease, subject's medical history, subject's health condition, subject degree of response to drugs, etc.) to determine the therapeutically effective dose of the present invention.
  • inhibitor refers to a decrease in the baseline activity of a biological activity or process.
  • kit used in this article refers to a box used to contain chemical reagents for detecting chemical components, drug residues, virus types, etc.
  • the kit of the present invention may include (i) one, two or three types of FAK inhibitors, topoisomerase inhibitors and immune checkpoint inhibitors; and (ii) instructions, which indicate that Tumors are treated in subjects using FAK inhibitors, topoisomerase inhibitors, and immune checkpoint inhibitors.
  • the kit includes (i) a FAK inhibitor; and (ii) instructions indicating that the FAK inhibitor, topoisomerase inhibitor, and immune checkpoint inhibitor can be used to treat in a subject Tumor.
  • the kit includes (i) a topoisomerase inhibitor; and (ii) instructions indicating that the FAK inhibitor, topoisomerase inhibitor, and immune checkpoint inhibitor can be used in Treat tumors in subjects.
  • the kit includes (i) an immune checkpoint inhibitor; and (ii) instructions indicating that a FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor can be used to treat a disease in a subject. in the treatment of tumors.
  • the kit includes (i) a FAK inhibitor, a topoisomerase inhibitor and an immune checkpoint inhibitor; and (ii) instructions indicating that the FAK inhibitor, topoisomerase inhibitor can be used inhibitors and immune checkpoint inhibitors to treat tumors in subjects.
  • the kit includes (i) a FAK inhibitor; and (ii) instructions indicating that the FAK inhibitor and the topoisomerase inhibitor can be used to treat a tumor in a subject. In one embodiment, the kit includes (i) a topoisomerase inhibitor; and (ii) instructions indicating that the topoisomerase inhibitor and the FAK inhibitor can be used to treat a tumor in a subject.
  • kits may include a first container including at least one dose of a drug including a FAK inhibitor, a second container including at least one dose of a topoisomerase inhibitor, and a package insert.
  • the third container includes at least one dose of an immune checkpoint inhibitor drug, and the package insert includes instructions for using the drug to treat a tumor in the subject.
  • the first container, the second container, and the third container may contain the same or different shapes (eg, vials, syringes, and bottles) and/or materials (eg, plastic or glass). Kits may also include other materials that may aid in administering the medication, such as diluents, filters, IV bags and tubing, needles, and syringes.
  • FAK inhibitor topoisomerase inhibitor
  • immune checkpoint inhibitor an effective amount also depends on the degree, severity and type of cell proliferation. A technician will be able to determine the appropriate dosage based on these and other factors.
  • FAK inhibitors, topoisomerase inhibitors, or immune checkpoint inhibitors can be administered in a suitable manner, such as oral, intravenous, intramuscular, or subcutaneous administration.
  • the drug may be administered orally with a pharmaceutically acceptable carrier such as an inert diluent or an absorbable edible carrier. They can be enclosed in hard- or soft-shell gelatin capsules, compressed into tablets, or can be mixed directly with the patient's food.
  • a pharmaceutically acceptable carrier such as an inert diluent or an absorbable edible carrier. They can be enclosed in hard- or soft-shell gelatin capsules, compressed into tablets, or can be mixed directly with the patient's food.
  • the drug may be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, lozenges, capsules, elixirs, suspensions, syrups, or wafers. Tablets, lozenges, pills, capsules, etc.
  • a binder such as tragacanth, acacia, corn starch or gelatin
  • an excipient such as dicalcium phosphate
  • a disintegrant such as corn starch, potato starch , alginic acid, etc.
  • lubricants such as magnesium stearate
  • sweeteners such as sucrose, fructose, lactose or aspartame; or flavoring agents.
  • solutions of the drug may be prepared in water, optionally mixed with a nontoxic surfactant.
  • Exemplary pharmaceutical dosage forms for injection or infusion include sterile aqueous solutions, dispersions, or sterile powders containing the active ingredient suitable for the extemporaneous preparation of sterile injection or infusion solutions or dispersions. Regardless, the final dosage form should be sterile, fluid, and stable under the conditions of manufacture and storage.
  • Sterile injectable solutions can be prepared by incorporating the required amount of the drug in an appropriate solvent with various other ingredients required from above, followed by filtered sterilization.
  • the preferred methods of preparation may be vacuum drying and freeze-drying techniques, which produce a powder of the active ingredient plus any other desired ingredients present after previous sterile filtration.
  • the amount of FAK inhibitor, topoisomerase inhibitor, or immune checkpoint inhibitor required for treatment may vary not only with the specific agent selected, but also with the route of administration, the nature of the disease being treated, and the patient's condition. Varies with age and condition and can ultimately be determined at the discretion of the attending physician or clinician. In general, however, dosages may range from about 0.1 to about 50 mg/kg of body weight per day.
  • the FAK inhibitors are administered at doses ranging from 5 mg/day to 300 mg/day in adults.
  • IN10018 or a pharmaceutically acceptable salt thereof is administered at a dosage of 5 mg/day to 100 mg/day in adults, for example, IN10018 or a pharmaceutically acceptable salt thereof is administered at a dosage of 25 mg/day in adults to 100 mg/day in adults.
  • a dose of 100 mg/day was administered based on IN10018.
  • the topoisomerase inhibitors are administered in a dose range of 1-300 mg/ m per week in adults.
  • irinotecan or a pharmaceutically acceptable salt thereof is administered at a dose of 1-300 mg/m per week in adults, based on irinotecan; etoposide or a pharmaceutically acceptable salt thereof; Acceptable salts are administered at doses of 1 to 300 mg/m per week in adults based on etoposide; teniposide is administered at doses between 1 and 300 mg/m per week in adults based on Niposide meter.
  • the immune checkpoint inhibitor is administered at a dose of 2-10 mg/kg or 50-1200 mg in adults every 2 to 3 weeks. In a specific embodiment, the immune checkpoint inhibitor is administered at a dose of 3-10 mg/kg or 100-1200 mg in adults, once every 2 weeks to 3 weeks.
  • the present disclosure also discloses the following:
  • FAK inhibitors topoisomerase inhibitors and immune checkpoint inhibitors, methods for treating tumors in a subject, wherein the topoisomerase inhibitor is not an anthracycline.
  • topoisomerase inhibitor is a topoisomerase I inhibitor.
  • topoisomerase I inhibitor is topotecan, irinotecan , Belotecan (Belotecan), Cositecan (Cositecan), Exatecan (DX-8951), Indenoisoquinoline (Indenoisoquinoline), Indotecan (LMP-400), Indimitecan (LMP-776) ), Simmitecan, Gimatecan (ST1481), EC-112002, PLX-038 (NK012), AR-67, or pharmaceutically acceptable salts thereof.
  • the topoisomerase I inhibitor is topotecan, irinotecan , Belotecan (Belotecan), Cositecan (Cositecan), Exatecan (DX-8951), Indenoisoquinoline (Indenoisoquinoline), Indotecan (LMP-400), Indimitecan (LMP-776) ), Simmitecan, Gimatecan (ST1481), EC-112002, PLX-038 (NK012), AR-67, or pharmaceutically acceptable salts thereof
  • topoisomerase I inhibitor is irinotecan, or Its pharmaceutically acceptable salt.
  • topoisomerase inhibitor is a topoisomerase II inhibitor.
  • topoisomerase II inhibitor is Etoposide, Teniposide ), sabarubicin, or its pharmaceutically acceptable salt.
  • topoisomerase II inhibitor is etoposide (Etoposide), tenipol Teniposide, or its pharmaceutically acceptable salt.
  • the FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor according to any one of embodiments 1-8, wherein the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody, PD-1/PD-L1 small molecule inhibitors or TIGIT antibodies.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody
  • the anti-PD-1/PD-L1 antibody is pembrolizumab, tislelizumab, nivolumab, toripalimab ), Atezolizumab, durvalumab, Avelumab, Atezolizumab, Camrelizumab, Sintilimab Anti-(Sintilimab), cemiplimab (Cemiplimab), envolimab (envafolimab), BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.
  • the immune checkpoint inhibitor is a PD-1/PD-L1 small molecule inhibitor
  • the PD- 1/PD-L1 small molecule inhibitors are INCB-086550, Lazertinib, IMMH-010, CA-170, ABSK043 or RRx-001.
  • the immune checkpoint inhibitor is a TIGIT antibody
  • the TIGIT antibody For Ociperlimab (Ociperlimab/BGB-A1217), Vibostolimab (Vibostolimab), domvanalimab (AB154), Tiragolumab (Tiragolumab), Belrestotug, Etigilimab (Etigilimab), ONO -4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005 , ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.
  • the FAK inhibitor, the topoisomerase inhibitor and the immune checkpoint inhibitor according to any one of embodiments 1-14, the FAK inhibitor, the topoisomerase inhibitor and the immune checkpoint inhibitor Point inhibitors are administered to the subject simultaneously or sequentially.
  • the tumor is bladder cancer, breast cancer, cervical cancer, colon cancer (including Colorectal cancer), esophageal cancer, esophageal squamous cell carcinoma, head and neck cancer, liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer), melanoma, myeloma, rhabdomyosarcoma, inflammatory myofibroblastic tumor, neuroblastoma Cell tumors, pancreatic cancer, prostate cancer, kidney cancer, renal cell carcinoma, sarcoma (including osteosarcoma), skin cancer (including squamous cell carcinoma), gastric cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma , leiomyosarcoma, liposarcoma, nasopharyngeal carcinoma, neuroendocrine carcinoma, ovarian cancer, salivary
  • kits or pharmaceutically acceptable composition comprising:
  • kits or composition of embodiment 18, wherein the FAK inhibitor is IN10018, Defactinib, GSK2256098, PF-00562271, VS-4718, APG-2449, AMP945, AMP886 or pharmaceutically acceptable ones thereof
  • the salt is preferably IN10018, 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:
  • topoisomerase I inhibitor is Topotecan, Irinotecan, Belotecan, Corsi Cositecan, Exatecan (DX-8951), Indenoisoquinoline, Indotecan (LMP-400), Indimitecan (LMP-776), Simmitecan, Gimatecan (ST1481), EC-112002, PLX-038 (NK012), AR-67, or pharmaceutically acceptable salts thereof.
  • kits or composition of embodiment 24, wherein the topoisomerase II inhibitor is Etoposide, Teniposide, or a pharmaceutically acceptable salt thereof.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody
  • the anti-PD-1/PD -L1 antibodies are pembrolizumab, Tislelizumab, Nivolumab, Toripalimab, Atezolizumab, durvalumab, Avelumab, Atezolizumab, Camrelizumab, Sintilimab, Cimeplimab (Cemiplimab), envolimab (envafolimab), BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.
  • TIGIT antibody is Ociperlimab/BGB -A1217), Vibostolimab, domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686,
  • the above acceptable salt; the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or a PD-1/PD-L1 small molecule inhibitor, especially an anti-PD-1/PD-L1 antibody.
  • kits or composition of embodiment 33, wherein the tumor is breast cancer or colon cancer (including colorectal cancer).
  • a method of treating tumors in a subject comprising administering to the subject a therapeutically effective amount of a FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor, wherein the topoisomerase The inhibitor is not an anthracycline.
  • topoisomerase I inhibitor is Topotecan, Irinotecan, Belotecan, Cositecan ), Exatecan (DX-8951), Indenoisoquinoline (Indenoisoquinoline), Indotecan (LMP-400), Indimitecan (LMP-776), Simmitecan (Simmitecan), Gimatecan (ST1481), EC-112002, PLX-038(NK012), AR-67, or pharmaceutically acceptable salts thereof.
  • the topoisomerase I inhibitor is Topotecan, Irinotecan, Belotecan, Cositecan ), Exatecan (DX-8951), Indenoisoquinoline (Indenoisoquinoline), Indotecan (LMP-400), Indimitecan (LMP-776), Simmitecan (Simmitecan), Gimatecan (ST1481), EC-112002, PLX-038(NK012), AR-67, or pharmaceutically acceptable salts thereof.
  • topoisomerase I inhibitor is irinotecan, or a pharmaceutically acceptable salt thereof.
  • topoisomerase inhibitor is a topoisomerase II inhibitor.
  • topoisomerase II inhibitor is Etoposide, Teniposide, Sabarubicin, or a pharmaceutically acceptable compound thereof. Take that with a grain of salt.
  • topoisomerase II inhibitor is Etoposide, Teniposide, or a pharmaceutically acceptable salt thereof.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody, a PD-1/PD-L1 small molecule inhibitor, or a TIGIT antibody.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody
  • the anti-PD-1/PD-L1 antibody is pembrolizumab, tislelizumab, nivolumab, toripalimab, atezolizumab, durvalumab (durvalumab), Avelumab, Atezolizumab, Camrelizumab, Sintilimab, Cemiplimab, Envafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.
  • the immune checkpoint inhibitor is a PD-1/PD-L1 small molecule inhibitor
  • the PD- 1/PD-L1 small molecule inhibitors are INCB-086550, Lazertinib, IMMH-010, CA-170, ABSK043 or RRx-001.
  • the immune checkpoint inhibitor is a TIGIT antibody
  • the TIGIT antibody is Ociperlimab (Ociperlimab/BGB-A1217), Vibostolimab, domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof
  • the topoisomerase inhibitor is Irinotecan or a pharmaceutically acceptable salt thereof salt
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or a small molecule inhibitor of PD-1/PD-L1, especially an anti-PD-1/PD-L1 antibody.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof
  • the topoisomerase inhibitor is Etoposide, Teniposide ), or a pharmaceutically acceptable salt thereof
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or a PD-1/PD-L1 small molecule inhibitor, especially an anti-PD-1/PD- L1 antibodies.
  • the tumor is bladder cancer, breast cancer, cervical cancer, colon cancer (including colorectal cancer), esophageal cancer, esophageal squamous cell carcinoma, head and neck cancer Cancer, liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer), melanoma, myeloma, rhabdomyosarcoma, inflammatory myofibroblastic tumor, neuroblastoma, pancreatic cancer, prostate cancer, kidney cancer, renal cell carcinoma , Sarcoma (including osteosarcoma), skin cancer (including squamous cell carcinoma), gastric cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal cancer, neuroendocrine cancer, Ovarian cancer, salivary gland cancer, metastases from spindle cell carcinoma, anaplastic large
  • AML Acute myeloid leukemia
  • ALL acute lymphoblastic leukemia
  • DLBCL diffuse large B-cell lymphoma
  • FL follicular lymphoma
  • CLL chronic lymphocytic leukemia
  • CML chronic myelogenous leukemia
  • the tumor is breast cancer, ovarian cancer, colon cancer (including colorectal cancer), lung cancer (including small cell lung cancer and non-small cell lung cancer), melanoma or pancreatic cancer.
  • FAK inhibitors for use in methods of treating tumors by increasing immunogenic cell death in a subject, wherein the topoisomerase inhibitor is not anthracene Ring type.
  • the FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor of 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, the The structure of IN10018 is as follows:
  • topoisomerase I inhibitor is topotecan, irinotecan , Belotecan (Belotecan), Cositecan (Cositecan), Exatecan (DX-8951), Indenoisoquinoline (Indenoisoquinoline), Indotecan (LMP-400), Indimitecan (LMP-776) ), Simmitecan, Gimatecan (ST1481), EC-112002, PLX-038 (NK012), AR-67, or pharmaceutically acceptable salts thereof.
  • the topoisomerase I inhibitor is topotecan, irinotecan , Belotecan (Belotecan), Cositecan (Cositecan), Exatecan (DX-8951), Indenoisoquinoline (Indenoisoquinoline), Indotecan (LMP-400), Indimitecan (LMP-776) ), Simmitecan, Gimatecan (ST1481), EC-112002, PLX-038 (NK012), AR-67, or pharmaceutically acceptable salts thereof
  • topoisomerase II inhibitor is Etoposide, Teniposide ), sabarubicin, or its pharmaceutically acceptable salt.
  • topoisomerase II inhibitor is etoposide, tenipol Teniposide, or its pharmaceutically acceptable salt.
  • the anti-PD-1/PD-L1 antibody is pembrolizumab, tislelizumab, nivolumab, toripalimab ), Atezolizumab, durvalumab, Avelumab, Atezolizumab, Camrelizumab, Sintilimab Anti-(Sintilimab), cemiplimab (Cemiplimab), envolimab (envafolimab), BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.
  • the immune checkpoint inhibitor is a PD-1/PD-L1 small molecule inhibitor
  • the PD-1/PD-L1 small molecule inhibitor is INCB-086550, Lazertinib, IMH-010, CA-170, ABSK043 or RRx-001.
  • the FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor according to any one of embodiments 52-60, wherein the immune checkpoint inhibitor is a TIGIT antibody, further, the TIGIT antibody For Ociperlimab (Ociperlimab/BGB-A1217), Vibostolimab (Vibostolimab), domvanalimab (AB154), Tiragolumab (Tiragolumab), Belrestotug, Etigilimab (Etigilimab), ONO -4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005 , ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.
  • TIGIT antibody For Ociperlimab (Ociperlimab/BGB-
  • the FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor of embodiment 52 wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; the topoisomerase inhibitor The agent is irinotecan or a pharmaceutically acceptable salt thereof; the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or a small molecule inhibitor of PD-1/PD-L1, especially an anti-PD-1/PD-L1 antibody. PD-1/PD-L1 antibodies.
  • the FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor of embodiment 52 wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; the topoisomerase inhibitor The agent is Etoposide, Teniposide, or a pharmaceutically acceptable salt thereof; the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or PD-1/PD- L1 small molecule inhibitors, especially anti-PD-1/PD-L1 antibodies.
  • the FAK inhibitor, the topoisomerase inhibitor and the immune checkpoint inhibitor according to any one of embodiments 52-65, the FAK inhibitor, the topoisomerase inhibitor and the immune checkpoint inhibitor Point inhibitors are administered to the subject simultaneously or sequentially.
  • the tumor is bladder cancer, breast cancer, cervical cancer, colon cancer (including Colorectal cancer), esophageal cancer, esophageal squamous cell carcinoma, head and neck cancer, liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer), melanoma, myeloma, rhabdomyosarcoma, inflammatory myofibroblastic tumor, neuroblastoma Cell tumors, pancreatic cancer, prostate cancer, kidney cancer, renal cell carcinoma, sarcoma (including osteosarcoma), skin cancer (including squamous cell carcinoma), gastric cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma , leiomyosarcoma, liposarcoma, nasopharyngeal carcinoma, neuroendocrine carcinoma, ovarian cancer,
  • a method of treating tumors by increasing immunogenic cell death in a subject comprises administering to said subject a therapeutically effective amount of a FAK inhibitor, a topoisomerase inhibitor, and an immune checkpoint inhibitor , wherein the topoisomerase inhibitor is not an anthracycline.
  • topoisomerase inhibitor is a topoisomerase I inhibitor.
  • topoisomerase I inhibitor is Topotecan, Irinotecan, Belotecan, Co. Cositecan, Exatecan (DX-8951), Indenoisoquinoline, Indotecan (LMP-400), Indimitecan (LMP-776), Simmitecan, Gimatecan (ST1481), EC-112002, PLX-038 (NK012), AR-67, or pharmaceutically acceptable salts thereof.
  • topoisomerase I inhibitor is irinotecan, or a pharmaceutically acceptable salt thereof.
  • topoisomerase inhibitor is a topoisomerase II inhibitor.
  • topoisomerase II inhibitor is Etoposide, Teniposide, Sabarubicin, or a pharmaceutically acceptable compound thereof. Take that with a grain of salt.
  • topoisomerase II inhibitor is Etoposide, Teniposide, or a pharmaceutically acceptable salt thereof.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody, a PD-1/PD-L1 small molecule inhibitor, or a TIGIT antibody.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody
  • the anti-PD-1/PD-L1 antibody is pembrolizumab, tislelizumab, nivolumab, toripalimab, atezolizumab, durvalumab (durvalumab), Avelumab, Atezolizumab, Camrelizumab, Sintilimab, Cemiplimab, Envafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.
  • the immune checkpoint inhibitor is a PD-1/PD-L1 small molecule inhibitor
  • the PD- 1/PD-L1 small molecule inhibitors are INCB-086550, Lazertinib, IMMH-010, CA-170, ABSK043 or RRx-001.
  • the immune checkpoint inhibitor is a TIGIT antibody
  • the TIGIT antibody is Ociperlimab/BGB-A1217, Vibostolimab, domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof
  • the topoisomerase inhibitor is Irinotecan or a pharmaceutically acceptable salt thereof salt
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or a small molecule inhibitor of PD-1/PD-L1, especially an anti-PD-1/PD-L1 antibody.
  • the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof
  • the topoisomerase inhibitor is Etoposide, Teniposide ), or a pharmaceutically acceptable salt thereof
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or a PD-1/PD-L1 small molecule inhibitor, especially an anti-PD-1/PD- L1 antibodies.
  • the tumor is bladder cancer, breast cancer, cervical cancer, colon cancer (including colorectal cancer), esophageal cancer, esophageal squamous cell carcinoma, head and neck cancer Cancer, liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer), melanoma, myeloma, rhabdomyosarcoma, inflammatory myofibroblastic tumor, neuroblastoma, pancreatic cancer, prostate cancer, kidney cancer, renal cell carcinoma , Sarcoma (including osteosarcoma), skin cancer (including squamous cell carcinoma), gastric cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal cancer, neuroendocrine cancer, Ovarian cancer, salivary gland cancer, metastases from spindle cell carcinoma, ana
  • AML Acute myeloid leukemia
  • ALL acute lymphoblastic leukemia
  • DLBCL diffuse large B-cell lymphoma
  • FL follicular lymphoma
  • CLL chronic lymphocytic leukemia
  • CML chronic myelogenous leukemia
  • the tumor is breast cancer, ovarian cancer, colon cancer (including colorectal cancer), lung cancer (including small cell lung cancer and non-small cell lung cancer), melanoma or pancreatic cancer.
  • a FAK inhibitor in the preparation of a medicament for treating tumors in a subject, wherein the FAK inhibitor, topoisomerase inhibitor and immune checkpoint inhibitor are administered to the subject, wherein the topoisomerase inhibitor
  • the isomerase inhibitor is not an anthracycline.
  • a topoisomerase inhibitor in the preparation of a medicament for treating a tumor in a subject, wherein a FAK inhibitor, the topoisomerase inhibitor and an immune checkpoint inhibitor are administered to the subject, wherein The topoisomerase inhibitor is not an anthracycline.
  • an immune checkpoint inhibitor in the manufacture of a medicament for treating a tumor in a subject, wherein a FAK inhibitor, a topoisomerase inhibitor, and the immune checkpoint inhibitor are administered to the subject, wherein the The above topoisomerase inhibitors are not anthracyclines.
  • a FAK inhibitor a topoisomerase inhibitor and an immune checkpoint inhibitor in the preparation of a combination drug for treating tumors, wherein the topoisomerase inhibitor is not an anthracycline.
  • a FAK inhibitor in the preparation of a medicament for the treatment of tumors in combination with a topoisomerase inhibitor and an immune checkpoint inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.
  • a topoisomerase inhibitor in the preparation of a drug for the treatment of tumors in combination with a FAK inhibitor and an immune checkpoint inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.
  • an immune checkpoint inhibitor in the preparation of a medicament for the treatment of tumors in combination with a FAK inhibitor and a topoisomerase inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.
  • a FAK inhibitor a topoisomerase inhibitor and an immune checkpoint inhibitor in the preparation of a drug for combined treatment of tumors, wherein the topoisomerase inhibitor is not an anthracycline.
  • a FAK inhibitor for the preparation of a medicament for the treatment of tumors in combination with a topoisomerase inhibitor and an immune checkpoint inhibitor, wherein the topoisomerase inhibitor is not an anthracycline.
  • topoisomerase inhibitor wherein the topoisomerase inhibitor is not an anthracycline, in the preparation of a medicament for the treatment of tumors in combination with a FAK inhibitor and an immune checkpoint inhibitor.
  • the salt is 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:
  • topoisomerase inhibitor is a topoisomerase I inhibitor.
  • topoisomerase I inhibitor is Topotecan, Irinotecan, Belotecan, Cositecan ), Exatecan (DX-8951), Indenoisoquinoline (Indenoisoquinoline), Indotecan (LMP-400), Indimitecan (LMP-776), Simmitecan (Simmitecan), Gimatecan (ST1481), EC-112002, PLX-038(NK012), AR-67, or pharmaceutically acceptable salts thereof.
  • the topoisomerase I inhibitor is Topotecan, Irinotecan, Belotecan, Cositecan ), Exatecan (DX-8951), Indenoisoquinoline (Indenoisoquinoline), Indotecan (LMP-400), Indimitecan (LMP-776), Simmitecan (Simmitecan), Gimatecan (ST1481), EC-112002, PLX-038(NK012), AR-67, or pharmaceutically acceptable salts thereof.
  • topoisomerase I inhibitor is irinotecan or a pharmaceutically acceptable salt thereof.
  • topoisomerase inhibitor is a topoisomerase II inhibitor.
  • topoisomerase II inhibitor is Etoposide, Teniposide, Sabarubicin, or a pharmaceutically acceptable compound thereof. Take that with a grain of salt.
  • topoisomerase II inhibitor is Etoposide, Teniposide, or a pharmaceutically acceptable salt thereof.
  • any one of embodiments 86-103, wherein the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody, a PD-1/PD-L1 small molecule inhibitor, or a TIGIT antibody.
  • the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody
  • the anti-PD-1/PD-L1 antibody is pembrolizumab, tislelizumab, nivolumab, toripalimab, atezolizumab, durvalumab (durvalumab), Avelumab, Atezolizumab, Camrelizumab, Sintilimab, Cemiplimab, Envafolimab, BMS-936559, JS003, SHR-1316, GS-4224, AN-4005 or MX-10181.
  • the immune checkpoint inhibitor is a PD-1/PD-L1 small molecule inhibitor
  • the PD- 1/PD-L1 small molecule inhibitors are INCB-086550, Lazertinib, IMMH-010, CA-170, ABSK043 or RRx-001.
  • the immune checkpoint inhibitor is a TIGIT antibody
  • the TIGIT antibody is Ociperlimab/BGB-A1217, Vibostolimab, domvanalimab (AB154), Tiragolumab, Belrestotug, Etigilimab, ONO-4686, JS-006, AZD-2936, HLX-301, SEA-TGT, M-6223, IBI-939, COM-902, AB-308, AGEN-1777, AK-127, BAT-6021, BAT-6005, ASP-8374, PM-1022, BMS-986207, HB0036 or IBI-321.
  • any one of embodiments 86-96 wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; the topoisomerase inhibitor is Irinotecan or a pharmaceutically acceptable salt thereof.
  • Pharmaceutically acceptable salt; the immune checkpoint inhibitor is an anti-PD-1/PD-L1 antibody or a PD-1/PD-L1 small molecule inhibitor, especially an anti-PD-1/PD-L1 antibody.
  • any one of embodiments 86-96 wherein the FAK inhibitor is IN10018 or a pharmaceutically acceptable salt thereof; the topoisomerase inhibitor is Etoposide, Etoposide, or a pharmaceutically acceptable salt thereof.
  • any one of embodiments 86-109, the FAK inhibitor, the topoisomerase inhibitor and the immune checkpoint inhibitor are administered to the subject simultaneously or sequentially.
  • the tumor is bladder cancer, breast cancer, cervical cancer, colon cancer (including colorectal cancer), esophageal cancer, esophageal squamous cell carcinoma, head and neck cancer Cancer, liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer), melanoma, myeloma, rhabdomyosarcoma, inflammatory myofibroblastic tumor, neuroblastoma, pancreatic cancer, prostate cancer, kidney cancer, renal cell carcinoma , Sarcoma (including osteosarcoma), skin cancer (including squamous cell carcinoma), gastric cancer, testicular cancer, thyroid cancer, uterine cancer, mesothelioma, cholangiocarcinoma, leiomyosarcoma, liposarcoma, nasopharyngeal cancer, neuroendocrine cancer, Ovarian cancer, salivary gland cancer, metastases from spindle cell carcinoma, ana
  • AML Acute myeloid leukemia
  • ALL acute lymphoblastic leukemia
  • DLBCL diffuse large B-cell lymphoma
  • FL follicular lymphoma
  • CLL chronic lymphocytic leukemia
  • CML chronic myelogenous leukemia
  • the tumor is breast cancer, ovarian cancer, colon cancer (including colorectal cancer), lung cancer (including small cell lung cancer and non-small cell lung cancer), melanoma or pancreatic cancer.
  • Example 1 Study on FAK target inhibition to enhance breast cancer cell death in response to Etoposide immunogenic cell death
  • Experimental Antibody Recombinant Alexa 647 fluorescent Anti-Calreticulin (anti-calreticulin) antibody (Abcam, ab196159), Annexin V apoptosis detection kit (Invitrogen, A35110).
  • FAK siRNA is provided by Jima Gene, and the sequence is shown in the table below:
  • MDA-MB-231 cells were transfected with control siRNA or FAK siRNA at a final concentration of 50 nM respectively. After 24 hours of transfection, the cells were treated with 2 ⁇ M etoposide for 48 hours, and the cells were stained using Annexin V kit and detected by flow cytometry. The early and late apoptosis values of the cells were compared with the DMSO control group and plotted using Graphpad 8.0. The results showed that compared with the control group, apoptosis was significantly enhanced after etoposide treatment, as shown in Figure 1. Show.
  • MDA-MB-231 cells were transfected with control siRNA or FAK siRNA at a final concentration of 50 nM respectively. After 24 hours of transfection, the cells were treated with 2 ⁇ M etoposide for 48 hours, fluorescent staining was performed with Calreticulin antibody, and the staining results were analyzed by flow cytometry. FlowJo software results statistics show that after FAK silencing combined with etoposide, the release and exposure of Calreticulin were significantly enhanced, as shown in Figure 2.
  • Example 2 Study on FAK target inhibition to enhance breast cancer cell death in response to Teniposide immunogenic cell death
  • FAK siRNA is provided by Jima Gene, and the sequence is shown in the table below:
  • MDA-MB-231 cells were transfected with control siRNA or FAK siRNA at a final concentration of 50 nM respectively. 24 hours after transfection, 2 ⁇ M teniposide was treated for 48 hours, and the cells were stained using Annexin V kit and detected using flow cytometry. The early and late cell apoptosis values were counted and compared with the DMSO control group and plotted using Graphpad 8.0. The results showed that compared with the control group, apoptosis in the FAK silencing group was significantly enhanced after teniposide treatment, as shown in Figure 3.
  • MDA-MB-231 cells were transfected with control siRNA or FAK siRNA at a final concentration of 50 nM respectively. After 24 hours of transfection, the cells were treated with 2 ⁇ M teniposide for 48 hours, fluorescent staining was performed with Calreticulin antibody, and the staining results were analyzed by flow cytometry. Statistics from FlowJo software show that after FAK silencing combined with teniposide, the release and exposure of Calreticulin were significantly enhanced, as shown in Figure 4.
  • Example 3 Synergistic effect of IN10018 and etoposide in colon cancer CT26 cells
  • CT26 cells were cultured with RPMI 1640 (Shanghai Yuanpei, classification number: L210KJ, batch number: F210916) + 10% FBS (Gibco, classification number: 10099-141c, batch number: 2158737cp), and passaged twice.
  • RPMI 1640 Shanghai Yuanpei, classification number: L210KJ, batch number: F210916) + 10% FBS (Gibco, classification number: 10099-141c, batch number: 2158737cp)
  • the first concentration is 30 ⁇ M, 5-fold dilution, the last one is the control, the drug concentration is 0, and each drug concentration is set to 3 A composite hole.
  • another group of cells was established using the same drug concentration as described above. The difference is that 5 ⁇ M IN10018 is added to each well, the drugs are mixed, and incubated at 37°C in a 5% CO2 incubator for 72 hours.
  • Example 4 Study of IN10018 and etoposide in colon cancer CT26 cells
  • CT26 cells Institute of Cell Science, Chinese Academy of Sciences
  • RPMI 1640 Shanghai Yuanpei, classification number: L210KJ, batch number: F210916) + 10% FBS (Gibco, classification number: 10099-141c, batch number: 2158737cp)
  • FBS FBS
  • the third group was etoposide (source MCE, classification number: 33419-42-0 , batch number: 113741), the concentration is 20 ⁇ M, and the fourth group is the combination group of IN10018 (5 ⁇ M) and etoposide (20 ⁇ M). Mix the drugs and incubate in a 5% CO2 incubator at 37°C for 48 hours.
  • Annexin-V-FITC conjugate 195 ⁇ l Annexin-V-FITC conjugate, mix the cells gently, then add 5 ⁇ l Annexin-V-FITC antibody, gently Mix lightly.
  • Add 10 ⁇ l PI dye mix gently, incubate at room temperature in the dark for 15 minutes, and then analyze on a flow cytometer.
  • Example 5 Study on the induction of immunogenic cell death targets in mouse colon cancer CT26 cells by irinotecan and IN10018 in vitro
  • CT26 cells were maintained and passaged by Yingshi Biotechnology (Nanjing) Co., Ltd.
  • the cells were cultured in a monolayer in vitro.
  • the cell culture conditions were RPMI1640 medium plus 10% fetal calf serum and cultured in a 37°C, 5% CO2 incubator. Perform routine digestion pass-through with trypsin two to three times a week. When the cells are in the exponential growth phase and the adhesion confluence reaches 80%-90%, the cells are harvested and plated.
  • CT26 cells were digested with trypsin and collected and counted. According to the counting results, the cells were diluted with the corresponding complete culture medium to a dilution concentration of 50,000 cells per ml, and then plated on a 24-well cell culture plate. Each well was plated. 1ml of cell suspension equals 50,000 cells. After plating, the cells were cultured in a 37°C, 5% CO2 incubator.
  • the compounds to be tested irinotecan and IN10018, were added to different wells respectively.
  • the compounds to be tested were uniformly prepared and aliquoted, dissolved in DMSO, and the aliquot volume was 50 ⁇ L/tube, and placed at -20°C. Stored under light, one of them was taken out for this experiment for dosing and testing.
  • the groupings and drug concentrations are shown in Table 4.
  • Example 6 Study on the inhibitory activity of Teniposide and IN10018 on mouse colon cancer CT26 cells in vitro
  • the main reagent information is shown in Table 7.
  • CT26 cells were revived and maintained for passage by Yingshi Biotechnology (Nanjing) Co., Ltd.
  • the cells were cultured in monolayer adhesion in vitro, and the culture conditions were RPMI1640 medium plus 10% fetal bovine serum, 37°C, and 5% CO 2 . Passage cells using trypsin two to three times a week. When the cells are in the exponential growth phase and the adhesion confluence reaches 80%-90%, the cells are harvested and plated.
  • CT26 cells were digested with trypsin and collected and counted. According to the counting results, the cells were diluted to 30,000 cells per ml using RPMI1640+10% FBS, and then cells were plated on a 96-well cell flat-bottom culture plate, and added to each well. 0.1ml of cell suspension equals 3,000 cells. After cell plating is completed, place the culture plate in a 37°C, 5% CO2 incubator to continue culturing.
  • the compound to be tested teniposide
  • the compound to be tested was uniformly prepared and packaged before the experiment. Briefly, DMSO was used to dissolve the drug to 10mM, and the aliquot volume was 50 ⁇ L/tube. Store it at -20°C in the dark. In this experiment, one tube was taken out to add medicine and process the cells. Grouping and drug concentration information are shown in Table 8.
  • Teniposide was designed with 10 drug effect concentrations, of which the last drug effect concentration was 0 and was set as the negative control group.
  • Inhibition percentage ⁇ [A(0 Added Drug)-A(Blank)]--[A(Added Drug)-A(Blank)] ⁇ /[A(0 Added Drug)-A(Blank)] ⁇ 100%
  • Example 7 Study on the induction of immunogenic cell death targets by teniposide and IN10018 in mouse colon cancer CT26 cells in vitro
  • CT26 cells were maintained and passaged by Yingshi Biotechnology (Nanjing) Co., Ltd.
  • the cells were cultured in a monolayer in vitro.
  • the culture conditions were RPMI1640 medium plus 10% fetal bovine serum and cultured in a 37°C, 5% CO 2 incubator. Perform routine digestion pass-through with trypsin two to three times a week. When the cells are in the exponential growth phase and the adhesion confluence reaches 80%-90%, the cells are harvested and plated.
  • CT26 cells were digested with trypsin and collected and counted. According to the counting results, the cells were diluted with RPMI1640+10% FBS. The dilution concentration was 50,000 cells per ml, and then plated on a 12-well cell culture plate. Each well was Plate 2 ml of cell suspension or 100,000 cells.
  • the cells were cultured in a 37°C, 5% CO2 incubator.
  • the compound to be tested teniposide
  • the compound to be tested was uniformly prepared and aliquoted, and dissolved in DMSO.
  • the concentration was 10mM, and the aliquot volume was 50 ⁇ L/tube. Store in the dark at -20°C. In this experiment, one of the tubes was taken out for dosing and testing.
  • the groupings and drug concentrations are shown in Table 9.
  • the cells were photographed using a microscope, and then the cells were trypsinized and collected for flow cytometry staining.
  • This experiment evaluated the effect of teniposide alone and in combination with IN10018 on inducing the expression of immune cell death targets in CT26 cells under in vitro conditions.
  • Example 8 Study on the in vivo anti-tumor efficacy of the compatibility treatment between etoposide, IN10018 and anti-mouse PD-L1 antibody in colon cancer MC38 cell C57BL/6 mouse subcutaneous allograft tumor model
  • Table 12 In vivo drug efficacy experimental animal grouping and dosage regimen Note: 1.N: number of mice in each group; 2. Dosing volume: According to the mouse body weight 10mL/kg, if the body weight decreases by more than 15%, the animal will stop administration; until the body weight recovers to a decrease of 10%, Resume administration; 3. The dose of etoposide will be increased to 6 mg/kg on the 6th day after group administration;
  • Colorectal cancer cell MC38 (sourced from Nanjing Kebai Biotechnology Co., Ltd., product number: CBP60825) was maintained and passaged by Yingshi Biotechnology (Nanjing) Co., Ltd.
  • the cells were cultured in a monolayer in vitro.
  • the culture conditions were DMEM medium plus 10% fetal calf serum at 37°C and a 5% CO 2 incubator. Perform routine digestion with trypsin-EDTA for passage two to three times a week. When the cells are in the exponential growth phase and the saturation is 80%-90%, collect the cells, count them and inoculate them.
  • the animal's health condition continues to deteriorate, or the tumor volume exceeds 3,000mm 3 , or it has severe disease or pain, it must be euthanized. If the following conditions occur, the veterinarian will be notified and euthanasia will be implemented: obvious weight loss, weight loss of more than 20%; unable to freely eat and drink; the average tumor volume in the control group reaches 3,000mm 3 , and the experiment will be terminated.
  • the animals showed the following clinical manifestations and continued to worsen: piloerection, arched back, white ears, nose, eyes or feet, rapid breathing, convulsions, continuous diarrhea, dehydration, slow movement and vocalization.
  • TGI (%) which reflects the tumor growth inhibition rate.
  • TGI (%) [1-(Average tumor volume of a certain administration group-Average tumor volume of the administration group at the beginning of treatment)/(Average tumor volume of the solvent control group-Average tumor volume of the solvent control group at the beginning of treatment)] ⁇ 100%.
  • tumor growth was observed every day.
  • the tumors were divided into groups based on tumor volume.
  • the average tumor volume in the group was approximately 50 mm 3 . Due to tumor burden, the control group was euthanized on the 26th day after vaccination, that is, the 17th day after group administration, and the entire experiment ended.
  • the tumor volume in the control group was 2670.9 ⁇ 1438.9mm 3 .
  • the tumor volumes of the PD-L1 antibody (5mg/kg) and etoposide (3mg/kg) monotherapy groups were 2677.7 ⁇ 2029.0mm 3 and 2420.2 ⁇ 1377.3mm 3 respectively; etoposide + PD-L1 antibody (3+
  • the tumor volumes of the two drug combination treatment groups were 1947.6 ⁇ 539.7mm 3 and 1865.0 ⁇ 839.9mm 3 respectively; etoposide + IN10018 + PD-L1 antibody
  • the tumor volumes in the (3+25+5mg/kg) three-drug combination treatment group were 1245.6 ⁇ 1064.7mm 3 respectively.
  • the tumor inhibition rate TGI of the etoposide + IN10018 + PD-L1 antibody (3+25+5mg/kg) three-drug combination treatment group was 54.4% (p ⁇ 0.0001) .
  • the comprehensive tumor volume was compared with the three-drug combination group of etoposide + IN10018 + PD-L1 antibody (3 + 25 + 5 mg/kg) for statistical analysis.
  • the control group, PD-L1 antibody (5 mg/kg) and etoposide were analyzed statistically.
  • the tumor volumes of each dose group at different time periods are shown in Figure 11.
  • the average tumor volume of the PD-L1 antibody (5 mg/kg) and etoposide (3 mg/kg) monotherapy group was very close to that of the control group, and did not show any effect on tumor growth. It has inhibitory effect; while the two-drug combination treatment group of etoposide + PD-L1 antibody (3 + 5 mg/kg) and etoposide + IN10018 (3 + 25 mg/kg) and etoposide + IN10018 + PD-L1 antibody (3+25+5mg/kg)
  • the three-drug combination group all had significant tumor growth inhibition effects, and there were statistical differences compared with the control group.
  • the tumor volume of the three-drug combination group of etoposide + IN10018 + PD-L1 antibody (3 + 25 + 5 mg/kg) has always been smaller than that of the other treatment groups, and is similar to that of the PD-L1 antibody.
  • the experiment was conducted in accordance with the dosage regimen. During the experiment, the animals were observed for eating, drinking and other activities every day, and the animal weight was recorded three times a week. After 17 days of group administration, the average weight of the control group changed from 18.7g on the day of group administration (Day0) to 23.9g, with a weight growth rate of 28.8%; PD-L1 antibody (5mg/kg) and etoposide ( 3mg/kg) The average body weight of the monotherapy group changed from 18.8g and 19.1g on Day0 to 23.6g and 22.8g on Day17, respectively, and the weight change rates were 25.8% and 19.3% respectively; etoposide + PD-L1 antibody The average body weight of the two-drug combination treatment group (3+5mg/kg) and etoposide+IN10018 (3+25mg/kg) changed from 19.1g and 18.9g on Day0 to 22.3g and 22.5g on Day17, respectively.
  • Example 9 Study on the induction of immunogenic cell death target in mouse breast cancer 4T1 cells by Etoposide and AMP945 in vitro
  • AMP945 is provided by MCE, Lot No.:143253
  • 4T1 cells (from Nanjing Kebai Biotechnology Co., Ltd., Cat. No.: CBP60352) were grown with RPMI1640 (Shanghai Yuanpei, Cat No.: L210KJ, Lot No.: F210916) + 10% FBS (Gibco, Cat No.: 10099-141c, Lot No.: 2158737cp), the culture conditions were 37°C and 5% CO 2 . Perform routine digestion pass-through with trypsin two to three times a week. When the cells are in the exponential growth phase and the adhesion confluence reaches 80%-90%, the cells are harvested and plated. 4T1 cells were digested with trypsin and collected and counted.
  • the cells were diluted with RPMI1640+10% FBS.
  • the dilution concentration was 50,000 cells per ml, and then plated on a 12-well cell culture plate. Each well was Plate 2 ml of cell suspension or 100,000 cells. After plating, the cells were cultured in a 37°C, 5% CO2 incubator. After the cells were spread for 24 hours, six groups were set up. The first group was the control group and culture medium was added. The second group was AMP945 at a concentration of 3 ⁇ M. The third group was AMP945 at a concentration of 6 ⁇ M. The fourth group was Etoposide at a concentration of 5 ⁇ M.
  • the fifth group is the combination of AMP945 (3 ⁇ M) and Etoposide (5 ⁇ M)
  • the sixth group is the combination of AMP945 (6 ⁇ M) and Etoposide (5 ⁇ M).
  • Example 10 Study on the induction of immunogenic cell death target in mouse breast cancer 4T1 cells by irinotecan and AMP945 in vitro
  • Irinotecan is provided by Shanghai Chaolan Chemical Technology Center, Lot No.:202110
  • AMP945 is provided by MCE, Lot No.:143253
  • 4T1 cells (from Nanjing Kebai Biotechnology Co., Ltd., Cat. No.: CBP60352) were grown with RPMI1640 (Shanghai Yuanpei, Cat No.: L210KJ, Lot No.: F210916) + 10% FBS (Gibco, Cat No.: 10099-141c, Lot No.: 2158737cp), and the culture conditions were 37°C and 5% CO 2 . Perform routine digestion passaging with trypsin two to three times a week. When the cells are in the exponential growth phase and the adhesion confluence reaches 80%-90%, the cells are harvested and plated. 4T1 cells were digested with trypsin and collected and counted.
  • the cells were diluted with RPMI1640+10% FBS.
  • the dilution concentration was 50,000 cells per ml, and then plated on a 12-well cell culture plate. Each well was Plate 2 ml of cell suspension, which is 100,000 cells. After plating, the cells were cultured in a 37°C, 5% CO2 incubator. After the cells were spread for 24 hours, six groups were set up. The first group was the control group and culture medium was added. The second group was AMP945 with a concentration of 3 ⁇ M. The third group was AMP945 with a concentration of 6 ⁇ M. The fourth group was Irinotecan with a concentration of 120 ⁇ M.
  • the fifth group is the combination of AMP945 (3 ⁇ M) and Irinotecan (120 ⁇ M), and the sixth group is the combination of AMP945 (6 ⁇ M) and Irinotecan (120 ⁇ M). Mix the drugs and incubate at 37°C in a 5% CO2 incubator for 48 hours.

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Abstract

本公开涉及FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂联合治疗肿瘤。本公开还涉及FAK抑制剂和拓扑异构酶抑制剂联合治疗肿瘤。

Description

FAK抑制剂及拓扑异构酶抑制剂的药物组合及用途
本申请要求于2022年9月16日递交的中国专利申请202211129639.0和于2023年9月7日递交的中国专利申请202311155482.3的优先权，在此全文引用上述中国专利申请公开的内容以作为本申请的一部分。
技术领域
本发明属于药物化学领域。具体地，本发明涉及粘着斑激酶(Focal Adhesion Kinase,FAK)抑制剂与其它药物联用治疗肿瘤。
背景技术
肿瘤是威胁人们健康的第二大杀手。免疫原性细胞死亡(Immunogenic cell death，ICD)是基于细胞程序性死亡的叠加效应。癌细胞接触化疗或者靶向治疗类药物的同时可能激活细胞内部的压力信号，这一类压力信号包含内质网压力(ER stress)和活性氧压力(oxidative stress)。在压力信号的作用下细胞会先尝试修复压力，如果压力造成的损伤超出了修复的能力，细胞会启动程序性死亡过程。在这个过程中往往伴随着一类叫做损伤相关分子(Damage associated molecular patterns，DAMPs)的释放。这类DAMPs会特异性的被机体内抗原呈递细胞(Antigen-presenting cells，APC)上的模式识别受体识别，诱导APC成熟、分化和激活，并逐级呈递给效应T细胞等免疫细胞，从而使免疫细胞产生抗原记忆，再次发现相同来源的肿瘤细胞时免疫细胞就会特异性识别并杀伤肿瘤细胞。ICD启动的新的针对肿瘤的特异性免疫反应可增加对免疫检查点抑制剂(Immuno-check point inhibitor，ICI)的敏感，以此增效免疫检查点类抑制剂的作用，并可产生具有免疫记忆持久的抗肿瘤的反应。
FAK，又称为蛋白酪氨酸激酶2(PTK2)，是一种非受体酪氨酸激酶，并且是粘着斑复合体的关键组分。FAK在介导整合素和生长因子信号以调节肿瘤细胞的侵袭、增殖和存活方面发挥着重要作用。
拓扑异构酶的体内功能是通过切割DNA链释放DNA复制及转录过程中产生的张力，去除复制又前端产生的正超螺旋和转录时RNA聚合于游所产生的负超螺旋，再将切割形成的断端连接成完整DNA链，保证反应顺利进行。拓扑异构酶抑制剂与相应异构酶易解离复合物结合并使之形成稳定复合物，阻止复制叉顺利通过而至DNA链断裂，干扰DNA复制及转录。这类药物被广泛用于肿瘤的治疗。但是此类药物会在服用一段时间后产生一定的耐药。
化疗包括拓扑异构酶抑制剂和免疫治疗联合，已在临床中有的广泛的应用。但是，疗效仍有改善提高的空间，尤其是如何强化疗效，产生持久的抗肿瘤效果。
发明内容
本公开一方面提供了FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂在制备用于在对象中治疗肿瘤的药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂的药物组合产品，其用于在对象中治疗肿瘤，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了一种治疗肿瘤的方法，该方法包括向有需要的对象施用治疗有效量的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了一种试剂盒或药学上可接受的组合物，其包括：(a)FAK抑制剂；(b)拓扑异构酶抑制剂，所述拓扑异构酶抑制剂不为蒽环类；和(c)免疫检查点抑制剂。
本公开又一方面提供了FAK抑制剂和拓扑异构酶抑制剂在制备用于在治疗肿瘤的药物中的用途，其中所述FAK抑制剂用于增强所述拓扑异构酶抑制剂诱导的免疫原性细胞死亡，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了FAK抑制剂，其在治疗肿瘤中用于增强拓扑异构酶抑制剂诱导的免疫原性细胞死亡，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了一种治疗肿瘤的方法，该方法包括向有需要的对象施用治疗有效量的FAK抑制剂和拓扑异构酶抑制剂，其中所述FAK抑制剂用于增强所述拓扑异构酶抑制剂诱导的免疫原性细胞死亡，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂在制备用于联合治疗肿瘤的药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了FAK抑制剂在制备用于与拓扑异构酶抑制剂和免疫检查点抑制剂治疗肿瘤的联用药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了拓扑异构酶抑制剂在制备用于与FAK抑制剂和免疫检查点抑制剂治疗肿瘤的联用药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了免疫检查点抑制剂在制备用于与FAK抑制剂和拓扑异构酶抑制剂治疗肿瘤的联用药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了FAK抑制剂在制备用于与拓扑异构酶抑制剂和免疫检查点抑制剂联合治疗肿瘤的药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了拓扑异构酶抑制剂在制备用于与FAK抑制剂和免疫检查点抑制剂联合治疗肿瘤的药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了免疫检查点抑制剂在制备用于与FAK抑制剂和拓扑异构酶抑制剂联合治疗肿瘤的药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了一种试剂盒，其包括：FAK抑制剂；和说明书，该说明书指出该FAK抑制剂可用于与拓扑异构酶抑制剂和免疫检查点抑制剂联合治疗肿瘤，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了一种试剂盒，其包括：拓扑异构酶抑制剂；和说明书，该说明书指出该拓扑异构酶抑制剂可用于与FAK抑制剂和免疫检查点抑制剂联合治疗肿瘤，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了一种试剂盒，其包括：免疫检查点抑制剂；和说明书，该说明书指出该免疫检查点抑制剂可用于与FAK抑制剂和拓扑异构酶抑制剂联合治疗肿瘤，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开另一方面提供了一种治疗肿瘤的方法，该方法包括向有需要的对象施用治疗有效量的FAK抑制剂和拓扑异构酶抑制剂，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了FAK抑制剂和拓扑异构酶抑制剂的药物组合产品，其用于在有需要的对象中治疗肿瘤，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了FAK抑制剂和拓扑异构酶抑制剂在制备用于治疗肿瘤的联用药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了FAK抑制剂在制备用于与拓扑异构酶抑制剂治疗肿瘤的联用药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了拓扑异构酶抑制剂在制备用于与FAK抑制剂治疗肿瘤的联用药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了FAK抑制剂和拓扑异构酶抑制剂在制备用于联合治疗肿瘤的药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了FAK抑制剂在制备用于与拓扑异构酶抑制剂联合治疗肿瘤的药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了拓扑异构酶抑制剂在制备用于与FAK抑制剂联合治疗肿瘤的药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了一种试剂盒，其包括：FAK抑制剂；和说明书，该说明书指出该FAK抑制剂可用于与拓扑异构酶抑制剂联合治疗肿瘤，其中所述拓扑异构酶抑制剂不为蒽环类。
本公开又一方面提供了一种试剂盒，其包括：拓扑异构酶抑制剂；和说明书，该说明书指出该拓扑异构酶抑制剂可用于与FAK抑制剂联合治疗肿瘤，其中所述拓扑异构酶抑制剂不为蒽环类。
可选的，所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、APG-2449、AMP945、AMP886或其药学上可接受的盐，优选为IN10018、Defactinib、AMP945或其药学上可接受的盐，进一步优选为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐，所述IN10018结构如下：
所述Defactinib也称为地法替尼，CAS号为1345713-71-4；所述GSK2256098的CAS号为1224887-10-8。所述PF-00562271的CAS号为717907-75-0；所述VS-4718的CAS号为1061353-68-1；所述APG-2449为亚盛医药研发；所述AMP945的CAS号为1393653-34-3。
可选的，所述拓扑异构酶抑制剂为拓扑异构酶I抑制剂。
例如，所述拓扑异构酶I抑制剂为拓扑替康(Topotecan)、伊立替康(Irinotecan)、贝洛替康(Belotecan)、科西特康(Cositecan)、依喜替康(Exatecan, DX-8951)、茚并异喹啉(Indenoisoquinoline)、Indotecan(LMP-400)、Indimitecan(LMP-776)、希明替康(Simmitecan)、Gimatecan(ST1481)、EC-112002、PLX-038(NK012)、AR-67，或其药学上可接受的盐。
所述拓扑替康(Topotecan)的CAS号为123948-87-8；所述伊立替康(Irinotecan)的CAS号为97682-44-5；所述贝洛替康(Belotecan)的CAS号为256411-32-2；所述科西特康(Cositecan)的CAS号为401905-67-7；所述依喜替康(Exatecan，DX-8951)的CAS号为171335-80-1；所述茚并异喹啉(Indenoisoquinoline)的CAS号为97501-75-2；所述Indotecan(LMP-400)的CAS号为915303-09-2；所述Indimitecan(LMP-776)的CAS号为915360-05-3；所述希明替康(Simmitecan)的CAS号为951290-31-6；所述Gimatecan(ST1481)的CAS号为292618-32-7；所述EC-112002为Elucida Oncology Inc研发、所述PLX-038(NK012)的CAS号为86639-52-3；所述AR-67的CAS号为220913-32-6。
可选的，所述拓扑异构酶I抑制剂为伊立替康(Irinotecan)或其药学上可接受的盐。
可选的，所述拓扑异构酶抑制剂为拓扑异构酶II抑制剂。
例如，所述拓扑异构酶II抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)、莎巴比星(Sabarubicin)，或其药学上可接受的盐。
所述依托泊苷(Etoposide)的CAS号为33419-42-0；所述替尼泊苷(Teniposide)的CAS号为29767-20-2；所述莎巴比星(Sabarubicin)的CAS号为211100-13-9。
可选的，所述拓扑异构酶II抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)，或其药学上可接受的盐。
可选的，所述免疫检查点抑制剂为抗PD-1/PD-L1抗体、PD-1/PD-L1小分子抑制剂或者TIGIT抗体。
可选的，所述免疫检查点抑制剂为抗PD-1/PD-L1抗体，进一步的，所述抗PD-1/PD-L1抗体为帕博利珠单抗(pembrolizumab)、替雷利珠单抗(Tislelizumab)、尼伏单抗(Nivolumab)、特瑞普利单抗(Toripalimab)、阿替珠单抗(Atezolizumab)、度伐单抗(durvalumab)、阿维单抗(Avelumab)、阿特珠单抗(Atezolizumab)、卡瑞利珠单抗(Camrelizumab)、信迪利单抗(Sintilimab)、西米普利单抗(Cemiplimab)、恩沃利单抗(envafolimab)、BMS-936559、JS003、SHR-1316、GS-4224、AN-4005或者MX-10181。
可选的，所述免疫检查点抑制剂为PD-1/PD-L1小分子抑制剂，进一步的，所述PD-1/PD-L1小分子抑制剂为INCB-086550、拉泽替尼(Lazertinib)、IMMH-010、CA-170、ABSK043或者RRx-001。
可选的，所述免疫检查点抑制剂为TIGIT抗体，进一步的，所述TIGIT抗体为欧司珀利单抗(Ociperlimab/BGB-A1217)、维博利单抗(Vibostolimab)、domvanalimab(AB154)、替瑞利尤单抗(Tiragolumab)、Belrestotug、艾替利单抗(Etigilimab)、ONO-4686、JS-006、AZD-2936、HLX-301、SEA-TGT、M-6223、IBI-939、COM-902、AB-308、AGEN-1777、AK-127、BAT-6021、BAT-6005、ASP-8374、PM-1022、BMS-986207、HB0036或IBI-321。
可选的，所述FAK抑制剂为IN10018或其药学上可接受的盐，所述拓扑异构酶抑制剂为伊立替康(Irinotecan)或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂。
可选的，所述FAK抑制剂为IN10018或其药学上可接受的盐，所述拓扑异构酶抑制剂为伊立替康(Irinotecan)或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体。
可选的，所述FAK抑制剂为IN10018或其药学上可接受的盐，所述拓扑异构酶抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)，或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂。
可选的，所述FAK抑制剂为IN10018或其药学上可接受的盐，所述拓扑异构酶抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)，或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体。
可选的，所述FAK抑制剂、所述拓扑异构酶抑制剂和所述免疫检查点抑制剂被同时或依次施用于所述对象。
可选的，所述肿瘤为膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、神经胶质瘤或者恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)。
可选的，其中所述肿瘤为乳腺癌或结肠癌(包括结直肠癌)。
附图说明
为了更清楚地说明本公开实施例的技术方案，下面将对实施例的附图作简单地介绍，显而易见地，下面描述中的附图仅仅涉及本公开的一些实施例，而非对本发明的限制。
图1显示了FAK沉默后联合2μM依托泊苷处理48小时，增加了MDA-MB-231细胞凋亡。
图2显示了FAK沉默后联合2μM依托泊苷处理48小时，增强了Calreticulin释放及暴露。
图3显示了FAK沉默后联合2μM替尼泊苷处理48小时增加了MDA-MB-231细胞凋亡。
图4显示了FAK沉默后联合2μM替尼泊苷处理48小时，增强了Calreticulin释放及暴露。
图5显示了依托泊苷联合IN10018在结肠癌细胞CT26中的IC50值。
图6显示了结肠癌CT26细胞与药物孵育48小时后拍摄细胞的白光显微镜照片。
图7显示结肠癌CT26细胞与IN10018和依托泊苷孵育48小时后的情况，其中：7a显示了结肠癌CT26细胞与药物孵育48小时后CRT阳性细胞的百分比；7b显示了结肠癌CT26细胞与药物孵育48小时后Annexin V阳性细胞的百分比。
图8显示结肠癌CT26细胞与IN10018和伊立替康孵育48小时后的情况，其中：8a显示了结肠癌CT26细胞与药物孵育48小时后CRT阳性细胞的百分比；8b显示了结肠癌CT26细胞与药物孵育48小时后Annexin V阳性细胞的百分比。
图9显示了替尼泊苷及其联合IN10018对结肠癌CT26细胞的增殖的抑制作用相对于药物浓度的曲线。
图10显示结肠癌CT26细胞与IN10018和替尼泊苷孵育48小时后的情况，其中：10a显示了小鼠结肠癌细胞CT26细胞与药物孵育48小时后Calreticulin(CRT)阳性细胞百分比；10b显示了小鼠结肠癌细胞CT26细胞与药物孵48小时后Annexin-V阳性细胞百分比。
图11显示了MC38小鼠结直肠癌细胞C57BL/6小鼠皮下同种移植瘤模型给药后的肿瘤生长曲线，其中数据点代表组内平均肿瘤体积，误差线代表标准误(SEM)。
图12显示了MC38小鼠结直肠癌细胞C57BL/6小鼠皮下同种移植瘤模型给药后的体重变化曲线，其中数据点代表组内平均体重，误差线代表标准误(SEM)。
图13显示小鼠乳腺癌T41细胞与AMP945和Etoposide孵育48小时后的情况，其中：13a显示了小鼠乳腺癌T41细胞与药物孵育48小时后Calreticulin(CRT)阳性细胞百分比；13b显示了小鼠乳腺癌T41细胞与药物孵48小时后Annexin-V阳性细胞百分比。
图14显示小鼠乳腺癌T41细胞与AMP945和Irinotecan孵育48小时后的情况，其中：14a显示了小鼠乳腺癌T41细胞与药物孵育48小时后Calreticulin(CRT)阳性细胞百分比；14b显示了小鼠乳腺癌T41细胞与药物孵48小时后Annexin-V阳性细胞百分比。
具体实施方式
为使本公开实施例的目的、技术方案和优点更加清楚，下面将结合本公开实施例的附图，对本公开实施例的技术方案进行清楚、完整地描述。显然，所描述的实施例是本公开的一部分实施例，而不是全部的实施例。基于所描述的本公开的实施例，本领域普通技术人员在无需创造性劳动的前提下所获得的所有其他实施例，都属于本发明保护的范围。
本发明可在不偏离本发明基本属性的情况下以其它具体形式来实施。应该理解的是，在不冲突的前提下，本发明的任一和所有实施方案都可与任一其它实施方案或多个其它实施方案中的技术特征进行组合以得到另外的实施方案。本发明包括这样的组合得到的另外的实施方案。
本公开中提及的所有出版物和专利在此通过引用以它们的全部内容纳入本公开。如果通过引用纳入的任何出版物和专利中使用的用途或术语与本公开中使用的用途或术语冲突，那么以本公开的用途和术语为准。
本文所用的章节标题仅用于组织文章的目的，而不应被解释为对所述主题的限制。
除非另有规定，本文使用的所有技术术语和科学术语具有要求保护主题所属领域的通常含义。倘若对于某术语存在多个定义，则以本文定义为准。
除了在工作实施例中或另外指出之外，在说明书和权利要求中陈述的定量性质例如剂量的所有数字应理解为在所有情况中被术语“约”修饰。还应理解的是，本申请列举的任何数字范围意在包括该范围内的所有的子范围和该范围或子范围的各个端点的任何组合。
本公开中使用的“包括”、“含有”或者“包含”等类似的词语意指出现该词前面的要素涵盖出现在该词后面列举的要素及其等同，而不排除未记载的要素。本文所用的术语“含有”或“包括(包含)”可以是开放式、半封闭式和封闭式的。换言之，所述术语也包括“基本上由…组成”、或“由…组成”。
定义
本申请中所用的下列术语和符号具有如下所述的含义，其所处的上下文中另有说明的除外。本文所用的术语“FAK抑制剂”是指FAK的有效抑制剂，可适于哺乳动物，特别是人。在一些实施方案中，所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、AMP945、AMP886或其药学上可接受的盐，所述IN10018结构如下：
所述Defactinib也称为地法替尼，CAS号为1345713-71-4；所述GSK2256098的CAS号为1224887-10-8。所述PF-00562271的CAS号为717907-75-0；所述VS-4718的CAS号为1061353-68-1；所述APG-2449为亚盛医药研发；所述AMP945的CAS号为1393653-34-3。在一些实施方案中，所述FAK抑制剂优选为IN10018、Defactinib、AMP945或其药学上可接受的盐，在一些优选实施方案中，所述FAK抑制剂为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐。
本文所用术语“拓扑异构酶抑制剂”是指一类能够抑制DNA拓扑异构酶活性的化合物。常见的有拓扑异构酶I抑制剂和拓扑异构酶II抑制剂。
本文所用术语“蒽环类”是指一类拓扑异构酶II抑制剂，包括阿霉素、表阿霉素、柔红霉素和阿克拉霉素等。
本文所用的术语“免疫检查点抑制剂”是指能够通过调控免疫检查点通路(例如PD-1、TIGIT、CTLA-4、LAG-3、TIM-3等)来提高免疫系统活性的药物。在一些实施方案中，免疫检查点抑制剂为PD-1/PD-L1(程序性细胞死亡蛋白1)通路的拮抗剂(也称为“PD-1抑制剂”)或TIGIT抑制剂。PD-1抑制剂在本公开中也称为PD-1/PD-L1抑制剂。例如，在本公开的治疗方法、药物和用途中所述PD-1/PD-L1抑制剂为PD-1/PD-L1抗体，包括但不限于帕博利珠单抗(可瑞达/Keytruda/K药)、替雷利珠单抗(Tislelizumab/百泽安)、尼伏单抗(Nivolumab)、特瑞普利单抗(Toripalimab/拓益)、阿替珠单抗(Atezolizumab/泰圣奇)、度伐单抗(英飞凡/度伐鲁单抗/durvalumab)、阿维单抗(Avelumab/Bavencio)、阿特珠单抗(MPDL3280A/Atezolizumab/Tecentriq/T药)、BMS-936559(全人源抗PD-L1的IgG4单克隆抗体)、GS-4224、AN-4005或者MX-10181。在一些优选的实施方案中，PD-1抑制剂为特瑞普利单抗。在一些实施方案中，PD-1抑制剂用于治疗人对象。在一些实施方案中，PD-1为人PD-1。PD-1/PD-L1抑制剂也包括PD-1/PD-L1小分子抑制剂，例如INCB-086550、拉泽替尼(Lazertinib)、IMMH-010、CA-170、ABSK043或者RRx-001。TIGIT(也称为WUCAM、Vstm3、VSIG9)是Ig超家族的一种受体，是继PD-1/PD-L1之后的新型免疫检查点。例如，在本公开的治疗方法、药物和用途中，所述免疫检查点抑制剂为TIGIT抑制剂，包括但不限于欧司珀利单抗(Ociperlimab/BGB-A1217)、维博利单抗(Vibostolimab)、domvanalimab(AB154)、替瑞利尤单抗(Tiragolumab)、Belrestotug、艾替利单抗(Etigilimab)、ONO-4686、JS-006、AZD-2936、HLX-301、SEA-TGT、M-6223、IBI-939、COM-902、AB-308、AGEN-1777、AK-127、BAT-6021、BAT-6005、ASP-8374、PM-1022、BMS-986207、HB0036或IBI-321。在一些实施方案中，TIGIT抑制剂用于治疗人对象。为了避免歧义，本文中抗体均包括双抗。
本文所用的“药物组合”或者“药物组合产品”既可以指采用一个剂量单位形式的固定组合(例如所有药物活性成分以一种剂型存在)或者成套药盒以组合施用的产品的情形，也可以指一种药物与指示该药物可与另外的一种或多种药物联合使用的说明书的组合情形。
本文所用的“联合治疗”或者“联用药物”是指一种药物与另外的一种或多种药物联合使用来治疗疾病，既包括一种药物与另外的一种或多种药物的组合的情形，也包括一种药物与指示该药物可与另外的一种或多种药物联合使用的说明书的组合情形。
“同时或依次施用”在本申请中是指一个给药周期内(例如4周内、3周内、2周内、1周内或24小时以内)两种以上的药物同时或以一定时间间隔先后施用，药物施用的方式(例如口服、静脉、肌肉或皮下施用等)可以相同或不同，两种以上的药物的给药频率/周期可以相同或不同。当本公开的治疗方法、产品或用途涉及两种药物时，两种药物可同时或以一定时间间隔分别单独施用。当本公开的治疗方法、产品或用途涉及三种药物时，三种药物可以在同一时间点施用，或者两种药物在一个时间点施用而剩下一种药物在另一个时间点施用，或者所有三种药物各自在不同时间点施用。
在一些实施方案中，PD-1/PD-L1抑制剂被静脉(例如，作为静脉输注)或皮下施用或口服。优选地，PD-1/PD-L1抑制剂以静脉输注施用。
在一些实施方案中，TIGIT抑制剂被静脉(例如，作为静脉输注)或皮下施用或口服。优选地，TIGIT抑制剂以静脉输注施用。
免疫检查点抑制剂治疗癌症的能力取决于肿瘤组织内肿瘤抗原特异性T细胞的存在。这要求肿瘤组织表达将自身与其非转化的对应物区分开来的抗原，例如，通过被称为新抗原(neoantigen)的新型蛋白产物。肿瘤新抗原负荷与免疫原性和敏感性(例如，对检查点抑制剂疗法的敏感性)密切相关，这意味着免疫原性较差的肿瘤应该在很大程度上对这些药物耐药。用于释放可被APC摄取的肿瘤抗原的疗法，如诱导免疫原性细胞死亡(ICD)的那些，可能会促进有效的抗肿瘤免疫，特别是当进一步与检查点抑制剂联合时。
本文所用的术语“治疗”是指给患有疾病或者具有所述疾病的症状的对象施用一种或多种药物物质，用以治愈、缓解、减轻、改变、医治、改善、改进或影响所述疾病或者所述疾病的症状。在一些实施方案中，所述疾病是肿瘤或者癌症。
本文所用的术语“肿瘤”是指机体在各种致瘤因素的作用下，局部组织的细胞在基因水平上失去对其生长的正常调控，从而导致其克隆型异常增生而形成的异常病变。所述肿瘤包括，但不限于：膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、神经胶质瘤或者恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)。在一些实施方案中，所述肿瘤为乳腺癌或结肠癌(包括结直肠癌)。
本文所用的术语“对象”或“受试者”是指哺乳动物和非哺乳动物。哺乳动物是指哺乳类的任何成员，其包括但不限于：人；非人灵长类动物，如黑猩猩及其它猿类和猴类物种；农场动物，如牛、马、绵羊、山羊和猪；家畜，如兔、狗和猫；实验室动物，包括啮齿类动物，如大鼠、小鼠和豚鼠；等等。非哺乳动物的例子包括但不限于鸟等。术语“对象”并不限定特定的年龄或性别。在一些实施方案中，对象是人。
本文所用的术语“药学上可接受的”指的是无毒的、生物学上可耐受的，适合给对象施用的。
本文所用的术语“药学上可接受的盐”指的是无毒的、生物学上可耐受的适合给对象施用的酸加成盐，包括但不限于：与无机酸形成的酸加成盐，例如盐酸盐、氢溴酸盐、碳酸盐、碳酸氢盐、磷酸盐、硫酸盐、亚硫酸盐、硝酸盐等；以及与有机酸形成的酸加成盐，例如甲酸盐、乙酸盐、苹果酸盐、马来酸盐、富马酸盐、酒石酸盐、琥珀酸盐、柠檬酸盐、乳酸盐、甲磺酸盐、对甲苯磺酸盐、2-羟基乙磺酸盐、苯甲酸盐、水杨酸盐、硬脂酸盐和与式HOOC-(CH2)n-COOH(其中n是0-4)的链烷二羧酸形成的盐等。
此外，药学上可接受的酸加成盐可以按照由碱性化合物制备酸加成盐的常规操作通过将游离碱溶于合适的溶剂并且用酸处理该溶液来得到。本领域技术人员无需过多实验即可确定各种可用来制备无毒的药学上可接受的酸加成盐的合成方法。
本文所用的术语“药学上可接受的组合物”是指必须在化学和/或毒理学上与包括制剂的其他成分相容，和/或与接受其治疗的对象相容。本文所用的术语“治疗有效量”是指通常足以对对象产生有益治疗效果的量。可以通过常规方法(例如建模、剂量递增研究或临床试验)结合常规影响因素(例如给药方式、化合物的药代动力学、疾病的严重程度和病程、对象的病史、对象的健康状况、对象对药物的响应程度等)来确定本发明的治疗有效量。
本文所用的术语“抑制”是指生物活动或过程的基线活性的降低。
本文所用的术语“试剂盒”是指用于盛放检测化学成分、药物残留、病毒种类等化学试剂的盒子。本发明所述试剂盒可以是包括(i)FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂中的一种、二种或三种；以及(ii)说明书，所述说明书指出可使用FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂来在对象中治疗肿瘤。在一种实施方案中，试剂盒包括(i)FAK抑制剂；以及(ii)说明书，所述说明书指出可使用FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂来在对象中治疗肿瘤。在一种实施方案中，试剂盒包括(i)拓扑异构酶抑制剂；以及(ii)说明书，所述说明书指出可使用FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂来在对象中治疗肿瘤。在一种实施方案中，试剂盒包括(i)免疫检查点抑制剂；以及(ii)说明书，所述说明书指出可使用FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂来在对象中治疗肿瘤。在一种实施方案中，试剂盒包括(i)FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂；以及(ii)说明书，所述说明书指出可使用FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂来在对象中治疗肿瘤。在一种实施方案中，试剂盒包括(i)FAK抑制剂；以及(ii)说明书，所述说明书指出可使用FAK抑制剂和拓扑异构酶抑制剂来在对象中治疗肿瘤。在一种实施方案中，试剂盒包括(i)拓扑异构酶抑制剂；以及(ii)说明书，所述说明书指出可使用拓扑异构酶抑制剂和FAK抑制剂在对象中治疗肿瘤。
试剂盒的化合物可以包含在分开的容器中。可选地，两种或更多种化合物包含在同一容器中。例如，试剂盒可以包括第一容器、第二容器、第三容器和包装插页，其中第一容器包括至少一个剂量的包括FAK抑制剂的药物，第二容器包括至少一个剂量的拓扑异构酶抑制剂，第三容器包括至少一个剂量的免疫检查点抑制剂的药物，且所述包装插页包括使用药物治疗对象的肿瘤的说明。第一容器、第二容器和第三容器可以包含相同或不同形状(例如，小瓶、注射器和瓶)和/或材料(例如，塑料或玻璃)。试剂盒还可以包括可以有助于施用药物的其他材料，如稀释剂、过滤器、IV袋和管线、针和注射器。
给予受试者的FAK抑制剂、拓扑异构酶抑制剂、或免疫检查点抑制剂的精确量将取决于各种因素，例如给定的药物或化合物，药物制剂，给药途径，疾病类型，病症，所治疗的受试者或宿主的身份等，但是仍然可以由本领域技术人员常规确定。例如，确定有效量还取决于细胞增殖的程度，严重性和类型。技术人员将能够根据这些和其他因素确定合适的剂量。
FAK抑制剂、拓扑异构酶抑制剂、或免疫检查点抑制剂可选择合适的方式例如口服、静脉、肌肉或皮下施用给药。
例如，口服给药时，可以将药物与药学上可接受的载体例如惰性稀释剂或可吸收的食用载体一起口服给药。它们可以封装在硬壳或软壳明胶胶囊中，可以压制成片剂，或者可以直接与患者的食物混合。例如，药物可以与一种或多种赋形剂组合，并以可摄取的片剂，口腔片剂，锭剂，胶囊，酏剂，悬浮液，糖浆或糯米纸囊剂等形式使用。片剂，锭剂，丸剂，胶囊剂等可进一步包括：粘合剂，例如黄芪胶，阿拉伯胶，玉米淀粉或明胶；赋形剂，如磷酸二钙；崩解剂，例如玉米淀粉，马铃薯淀粉，海藻酸等；润滑剂，例如硬脂酸镁；或甜味剂，例如蔗糖，果糖，乳糖或阿斯巴甜；或调味剂。
例如，输注或注射静脉内或腹膜内给药时，药物的溶液可以在水中制备，任选地与无毒的表面活性剂混合。
用于注射或输注的示例性药物剂型包括：无菌水溶液，分散液，或包含活性成分的无菌粉末，该无菌粉末适合于临时制备无菌注射或输注溶液或分散液。无论如何，最终剂型在生产和储存条件下均应无菌，流动且稳定。
无菌注射溶液可以通过将所需量的药物与所需的上述各种其他成分掺入适当的溶剂中，然后过滤灭菌来制备。对于用于制备无菌注射溶液的无菌粉末，优选的制备方法可以是真空干燥和冷冻干燥技术，其可以产生活性成分加上先前无菌过滤后存在的任何其他所需成分的粉末。
用于治疗所需的FAK抑制剂、拓扑异构酶抑制剂、或免疫检查点抑制剂的量不仅可以随所选择的特定试剂而变化，还可以随给药途径，所治疗疾病的性质以及患者的年龄和状况而变化，并且最终可以由主治医师或临床医生自行决定。然而，一般而言，剂量可以在每天约0.1至约50mg/kg体重的范围内。
所述FAK抑制剂以成年人中5mg/天-300mg/天的剂量范围施用。在一种具体的实施方案中，IN10018或其药学上可接受盐以成年人中5mg/天-100mg/天的剂量施用，例如，IN10018或其药学上可接受盐以成年人中25mg/天-100mg/天的剂量施用，所述剂量以IN10018计。
所述拓扑异构酶抑制剂以成年人中每周1-300mg/m2的剂量范围施用。在一种具体的实施方式中，伊立替康或其药学上可接受的盐以成年人中每周1-300mg/m2的剂量施用，用量以伊立替康计；依托泊苷或其药学上可接受的盐以成年人中每周1-300mg/m2的剂量施用，用量以依托泊苷计；替尼泊苷以成年人中每周1-300mg/m2的剂量施用，用量以替尼泊苷计。
所述免疫检查点抑制剂每次给药以成年人中2-10mg/kg或者50-1200mg的剂量施用，每2周到3周给药一次。在一种具体的实施方式中，所述免疫检查点抑制剂每次给药以成年人中3-10mg/kg或者100-1200mg的剂量施用，每2周到3周给药一次。
本文所用的未具体定义的技术和科学术语具有本发明所属领域的技术人员通常理解的含义。
在一些实施方案中，本公开还公开了以下：
1.FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其用于在对象中治疗肿瘤的方法，其中所述拓扑异构酶抑制剂不为蒽环类。
2.如实施方案1所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、APG-2449、AMP945、AMP886或其药学上可接受的盐，优选为IN10018、Defactinib、AMP945或其药学上可接受的盐，进一步优选为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐，所述IN10018结构如下：
3.如实施方案1-2所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述拓扑异构酶抑制剂为拓扑异构酶I抑制剂。
4.如实施方案3所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述拓扑异构酶I抑制剂为拓扑替康(Topotecan)、伊立替康(Irinotecan)、贝洛替康(Belotecan)、科西特康(Cositecan)、依喜替康(Exatecan，DX-8951)、茚并异喹啉(Indenoisoquinoline)、Indotecan(LMP-400)、Indimitecan(LMP-776)、希明替康(Simmitecan)、Gimatecan(ST1481)、EC-112002、PLX-038(NK012)、AR-67，或其药学上可接受的盐。
5.如实施方案3-4任一项所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述拓扑异构酶I抑制剂为伊立替康(Irinotecan)，或其药学上可接受的盐。
6.如实施方案1-2任一项所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述拓扑异构酶抑制剂为拓扑异构酶II抑制剂。
7.如实施方案6所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述拓扑异构酶II抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)、莎巴比星(Sabarubicin)，或其药学上可接受的盐。
8.如实施方案7任一项所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述拓扑异构酶II抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)，或其药学上可接受的盐。
9.如实施方案1-8任一项所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体、PD-1/PD-L1小分子抑制剂或者TIGIT抗体。
10.如实施方案1-9任一项所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体，进一步的，所述抗PD-1/PD-L1抗体为帕博利珠单抗(pembrolizumab)、替雷利珠单抗(Tislelizumab)、尼伏单抗(Nivolumab)、特瑞普利单抗(Toripalimab)、阿替珠单抗(Atezolizumab)、度伐单抗(durvalumab)、阿维单抗(Avelumab)、阿特珠单抗(Atezolizumab)、卡瑞利珠单抗(Camrelizumab)、信迪利单抗(Sintilimab)、西米普利单抗(Cemiplimab)、恩沃利单抗(envafolimab)、BMS-936559、JS003、SHR-1316、GS-4224、AN-4005或者MX-10181。
11.如实施方案1-9任一项所述的用途、药物组合产品或者方法，其中所述免疫检查点抑制剂为PD-1/PD-L1小分子抑制剂，进一步的，所述PD-1/PD-L1小分子抑制剂为INCB-086550、拉泽替尼(Lazertinib)、IMMH-010、CA-170、ABSK043或者RRx-001。
12.如实施方案1-9任一项所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述免疫检查点抑制剂为TIGIT抗体，进一步的，所述TIGIT抗体为欧司珀利单抗(Ociperlimab/BGB-A1217)、维博利单抗(Vibostolimab)、domvanalimab(AB154)、替瑞利尤单抗(Tiragolumab)、Belrestotug、艾替利单抗(Etigilimab)、ONO-4686、JS-006、AZD-2936、HLX-301、SEA-TGT、M-6223、IBI-939、COM-902、AB-308、AGEN-1777、AK-127、BAT-6021、BAT-6005、ASP-8374、PM-1022、BMS-986207、HB0036或IBI-321。
13.如实施方案1所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述FAK抑制剂为IN10018或其药学上可接受的盐；所述拓扑异构酶抑制剂为伊立替康(Irinotecan)或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
14如实施方案1所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述FAK抑制剂为IN10018或其药学上可接受的盐；所述拓扑异构酶抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)，或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
15.如实施方案1-14任一项所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，所述FAK抑制剂、所述拓扑异构酶抑制剂和所述免疫检查点抑制剂被同时或依次施用于所述对象。
16.如实施方案1-15任一项所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述肿瘤为膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、神经胶质瘤或者恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)；优选地，所述肿瘤为乳腺癌、卵巢癌、结肠癌(包括结直肠癌)、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤或胰腺癌。
17.如实施方案1-16任一项所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述肿瘤为乳腺癌或结肠癌(包括结直肠癌)。
18.一种试剂盒或药学上可接受的组合物，其包括：
(a)FAK抑制剂；
(b)拓扑异构酶抑制剂，所述拓扑异构酶抑制剂不为蒽环类；和
(c)免疫检查点抑制剂。
19.如实施方案18所述的试剂盒或组合物，其中所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、APG-2449、AMP945、AMP886或其药学上可接受的盐，优选为IN10018、Defactinib、AMP945或其药学上可接受的盐，进一步优选为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐，所述IN10018结构如下：
20.如实施方案18-19所述的试剂盒或组合物，其中所述拓扑异构酶抑制剂为拓扑异构酶I抑制剂。
21.如实施方案20所述的试剂盒或组合物，其中所述拓扑异构酶I抑制剂为拓扑替康(Topotecan)、伊立替康(Irinotecan)、贝洛替康(Belotecan)、科西特康(Cositecan)、依喜替康(Exatecan，DX-8951)、茚并异喹啉(Indenoisoquinoline)、Indotecan(LMP-400)、Indimitecan(LMP-776)、希明替康(Simmitecan)、Gimatecan(ST1481)、EC-112002、PLX-038(NK012)、AR-67，或其药学上可接受的盐。
22.如实施方案20-21任一项所述的试剂盒或组合物，其中所述拓扑异构酶I抑制剂为伊立替康(Irinotecan)，或其药学上可接受的盐。
23.如实施方案18-19任一项所述的试剂盒或组合物，其中所述拓扑异构酶抑制剂为拓扑异构酶II抑制剂。
24.如实施方案23所述的试剂盒或组合物，其中所述拓扑异构酶II抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)、莎巴比星(Sabarubicin)，或其药学上可接受的盐。
25.如实施方案24所述的试剂盒或组合物，其中所述拓扑异构酶II抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)，或其药学上可接受的盐。
26.如实施方案18-25任一项所述的试剂盒或组合物，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体、PD-1/PD-L1小分子抑制剂或者TIGIT抗体。
27.如实施方案18-26任一项所述的试剂盒或组合物，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体，进一步的，所述抗PD-1/PD-L1抗体为帕博利珠单抗(pembrolizumab)、替雷利珠单抗(Tislelizumab)、尼伏单抗(Nivolumab)、特瑞普利单抗(Toripalimab)、阿替珠单抗(Atezolizumab)、度伐单抗(durvalumab)、阿维单抗(Avelumab)、阿特珠单抗(Atezolizumab)、卡瑞利珠单抗(Camrelizumab)、信迪利单抗(Sintilimab)、西米普利单抗(Cemiplimab)、恩沃利单抗(envafolimab)、BMS-936559、JS003、SHR-1316、GS-4224、AN-4005或者MX-10181。
28.如实施方案18-26任一项所述的用途、药物组合产品或者方法，其中所述免疫检查点抑制剂为PD-1/PD-L1小分子抑制剂，进一步的，所述PD-1/PD-L1小分子抑制剂为INCB-086550、拉泽替尼(Lazertinib)、IMMH-010、CA-170、ABSK043或者RRx-001。
29.如实施方案18-26任一项所述的试剂盒或组合物，其中所述免疫检查点抑制剂为TIGIT抗体，进一步的，所述TIGIT抗体为欧司珀利单抗(Ociperlimab/BGB-A1217)、维博利单抗(Vibostolimab)、domvanalimab(AB154)、替瑞利尤单抗(Tiragolumab)、Belrestotug、艾替利单抗(Etigilimab)、ONO-4686、JS-006、AZD-2936、HLX-301、SEA-TGT、M-6223、IBI-939、COM-902、AB-308、AGEN-1777、AK-127、BAT-6021、BAT-6005、ASP-8374、PM-1022、BMS-986207、HB0036或IBI-321。
30.如实施方案18所述的试剂盒或组合物，其中所述FAK抑制剂为IN10018或其药学上可接受的盐；所述拓扑异构酶抑制剂为伊立替康(Irinotecan)或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
31.如实施方案18所述的试剂盒或组合物，其中所述FAK抑制剂为IN10018或其药学上可接受的盐；所述拓扑异构酶抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)，或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
32.如实施方案18-31任一项所述的试剂盒或组合物，其中所述组合物用于药物。
33.如实施方案32所述的试剂盒或组合物，其中所述药物用于治疗肿瘤，所述肿瘤为膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、神经胶质瘤或者恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)；优选地，所述肿瘤为乳腺癌、卵巢癌、结肠癌(包括结直肠癌)、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤或胰腺癌。
34.如实施方案33所述的试剂盒或组合物，其中所述肿瘤为乳腺癌或结肠癌(包括结直肠癌)。
35.一种在对象中治疗肿瘤的方法，其中所述方法包括向所述对象施用治疗有效量的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述拓扑异构酶抑制剂不为蒽环类。
36.如实施方案35所述的方法，其中所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、APG-2449、AMP945、AMP886或其药学上可接受的盐，优选为IN10018、Defactinib、AMP945或其药学上可接受的盐，进一步优选为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐，所述IN10018结构如下：
37.如实施方案35-36所述的方法，其中所述拓扑异构酶抑制剂为拓扑异构酶I抑制剂。
38.如实施方案37所述的方法，其中所述拓扑异构酶I抑制剂为拓扑替康(Topotecan)、伊立替康(Irinotecan)、贝洛替康(Belotecan)、科西特康(Cositecan)、依喜替康(Exatecan，DX-8951)、茚并异喹啉(Indenoisoquinoline)、Indotecan(LMP-400)、Indimitecan(LMP-776)、希明替康(Simmitecan)、Gimatecan(ST1481)、EC-112002、PLX-038(NK012)、AR-67，或其药学上可接受的盐。
39.如实施方案38所述的方法，其中所述拓扑异构酶I抑制剂为伊立替康(Irinotecan)，或其药学上可接受的盐。
40.如实施方案35-36任一项所述的方法，其中所述拓扑异构酶抑制剂为拓扑异构酶II抑制剂。
41.如实施方案40所述的方法，其中所述拓扑异构酶II抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)、莎巴比星(Sabarubicin)，或其药学上可接受的盐。
42.如实施方案41所述的方法，其中所述拓扑异构酶II抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)，或其药学上可接受的盐。
43.如实施方案35-42任一项所述的方法，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体、PD-1/PD-L1小分子抑制剂或者TIGIT抗体。
44.如实施方案35-43任一项所述的方法，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体，进一步的，所述抗PD-1/PD-L1抗体为帕博利珠单抗(pembrolizumab)、替雷利珠单抗(Tislelizumab)、尼伏单抗(Nivolumab)、特瑞普利单抗(Toripalimab)、阿替珠单抗(Atezolizumab)、度伐单抗(durvalumab)、阿维单抗(Avelumab)、阿特珠单抗(Atezolizumab)、卡瑞利珠单抗(Camrelizumab)、信迪利单抗(Sintilimab)、西米普利单抗(Cemiplimab)、恩沃利单抗(envafolimab)、BMS-936559、JS003、SHR-1316、GS-4224、AN-4005或者MX-10181。
45.如实施方案35-43任一项所述的用途、药物组合产品或者方法，其中所述免疫检查点抑制剂为PD-1/PD-L1小分子抑制剂，进一步的，所述PD-1/PD-L1小分子抑制剂为INCB-086550、拉泽替尼(Lazertinib)、IMMH-010、CA-170、ABSK043或者RRx-001。
46.如实施方案35-43任一项所述的方法，其中所述免疫检查点抑制剂为TIGIT抗体，进一步的，所述TIGIT抗体为欧司珀利单抗(Ociperlimab/BGB-A1217)、维博利单抗(Vibostolimab)、domvanalimab(AB154)、替瑞利尤单抗(Tiragolumab)、Belrestotug、艾替利单抗(Etigilimab)、ONO-4686、JS-006、AZD-2936、HLX-301、SEA-TGT、M-6223、IBI-939、COM-902、AB-308、AGEN-1777、AK-127、BAT-6021、BAT-6005、ASP-8374、PM-1022、BMS-986207、HB0036或IBI-321。
47.如实施方案35所述的方法，其中所述FAK抑制剂为IN10018或其药学上可接受的盐；所述拓扑异构酶抑制剂为伊立替康(Irinotecan)或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
48.如实施方案35所述的方法，其中所述FAK抑制剂为IN10018或其药学上可接受的盐；所述拓扑异构酶抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)，或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
49.如实施方案35-48任一项所述的方法，所述FAK抑制剂、所述拓扑异构酶抑制剂和所述免疫检查点抑制剂被同时或依次施用于所述对象。
50.如实施方案35-49任一项所述的方法，其中所述肿瘤为膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、神经胶质瘤或者恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)；优选地，所述肿瘤为乳腺癌、卵巢癌、结肠癌(包括结直肠癌)、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤或胰腺癌。
51.如实施方案35-50任一项所述的方法，其中所述肿瘤为乳腺癌或结肠癌(包括结直肠癌)。
52.FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其用于通过在对象中增加免疫原性细胞死亡来治疗肿瘤的方法，其中所述拓扑异构酶抑制剂不为蒽环类。
53.如实施方案52所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、APG-2449、AMP945、AMP886或其药学上可接受的盐，优选为IN10018、Defactinib、AMP945或其药学上可接受的盐，进一步优选为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐，所述IN10018结构如下：
54.如实施方案52-53所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述拓扑异构酶抑制剂为拓扑异构酶I抑制剂。
55.如实施方案54所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述拓扑异构酶I抑制剂为拓扑替康(Topotecan)、伊立替康(Irinotecan)、贝洛替康(Belotecan)、科西特康(Cositecan)、依喜替康(Exatecan，DX-8951)、茚并异喹啉(Indenoisoquinoline)、Indotecan(LMP-400)、Indimitecan(LMP-776)、希明替康(Simmitecan)、Gimatecan(ST1481)、EC-112002、PLX-038(NK012)、AR-67，或其药学上可接受的盐。
56.如实施方案55所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述拓扑异构酶I抑制剂为伊立替康(Irinotecan)或其药学上可接受的盐。
57.如实施方案52-53任一项所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述拓扑异构酶抑制剂为拓扑异构酶II抑制剂。
58.如实施方案57所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述拓扑异构酶II抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)、莎巴比星(Sabarubicin)，或其药学上可接受的盐。
59.如实施方案58任一项所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述拓扑异构酶II抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)，或其药学上可接受的盐。
60.如实施方案52-59任一项所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体、PD-1/PD-L1小分子抑制剂或者TIGIT抗体。
61.如实施方案52-60任一项所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体，进一步的，所述抗PD-1/PD-L1抗体为帕博利珠单抗(pembrolizumab)、替雷利珠单抗(Tislelizumab)、尼伏单抗(Nivolumab)、特瑞普利单抗(Toripalimab)、阿替珠单抗(Atezolizumab)、度伐单抗(durvalumab)、阿维单抗(Avelumab)、阿特珠单抗(Atezolizumab)、卡瑞利珠单抗(Camrelizumab)、信迪利单抗(Sintilimab)、西米普利单抗(Cemiplimab)、恩沃利单抗(envafolimab)、BMS-936559、JS003、SHR-1316、GS-4224、AN-4005或者MX-10181。
62.如实施方案52-60任一项所述的用途、药物组合产品或者方法，其中所述免疫检查点抑制剂为PD-1/PD-L1小分子抑制剂，进一步的，所述PD-1/PD-L1小分子抑制剂为INCB-086550、拉泽替尼(Lazertinib)、IMMH-010、CA-170、ABSK043或者RRx-001。
63.如实施方案52-60任一项所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述免疫检查点抑制剂为TIGIT抗体，进一步的，所述TIGIT抗体为欧司珀利单抗(Ociperlimab/BGB-A1217)、维博利单抗(Vibostolimab)、domvanalimab(AB154)、替瑞利尤单抗(Tiragolumab)、Belrestotug、艾替利单抗(Etigilimab)、ONO-4686、JS-006、AZD-2936、HLX-301、SEA-TGT、M-6223、IBI-939、COM-902、AB-308、AGEN-1777、AK-127、BAT-6021、BAT-6005、ASP-8374、PM-1022、BMS-986207、HB0036或IBI-321。
64.如实施方案52所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述FAK抑制剂为IN10018或其药学上可接受的盐；所述拓扑异构酶抑制剂为伊立替康(Irinotecan)或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
65.如实施方案52所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查 点抑制剂，其中所述FAK抑制剂为IN10018或其药学上可接受的盐；所述拓扑异构酶抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)，或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
66.如实施方案52-65任一项所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，所述FAK抑制剂、所述拓扑异构酶抑制剂和所述免疫检查点抑制剂被同时或依次施用于所述对象。
67.如实施方案52-66任一项所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述肿瘤为膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、神经胶质瘤或者恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)；优选地，所述肿瘤为乳腺癌、卵巢癌、结肠癌(包括结直肠癌)、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤或胰腺癌。
68.如实施方案52-67任一项所述的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述肿瘤为乳腺癌或结肠癌(包括结直肠癌)。
69.一种通过在对象中增加免疫原性细胞死亡来治疗肿瘤的方法，其中所述方法包括向所述对象施用治疗有效量的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述拓扑异构酶抑制剂不为蒽环类。
70.如实施方案69所述的方法，其中所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、APG-2449、AMP945、AMP886或其药学上可接受的盐，优选为IN10018、Defactinib、AMP945或其药学上可接受的盐，进一步优选为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐，所述IN10018结构如下：
71.如实施方案69-70所述的方法，其中所述拓扑异构酶抑制剂为拓扑异构酶I抑制剂。
72.如实施方案69-71任一项所述的方法，其中所述拓扑异构酶I抑制剂为拓扑替康(Topotecan)、伊立替康(Irinotecan)、贝洛替康(Belotecan)、科西特康(Cositecan)、依喜替康(Exatecan，DX-8951)、茚并异喹啉(Indenoisoquinoline)、Indotecan(LMP-400)、Indimitecan(LMP-776)、希明替康(Simmitecan)、Gimatecan(ST1481)、EC-112002、PLX-038(NK012)、AR-67，或其药学上可接受的盐。
73.如实施方案72所述的方法，其中所述拓扑异构酶I抑制剂为伊立替康(Irinotecan)，或其药学上可接受的盐。
74.如实施方案69-70任一项所述的方法，其中所述拓扑异构酶抑制剂为拓扑异构酶II抑制剂。
75.如实施方案74所述的方法，其中所述拓扑异构酶II抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)、莎巴比星(Sabarubicin)，或其药学上可接受的盐。
76.如实施方案75所述的方法，其中所述拓扑异构酶II抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)，或其药学上可接受的盐。
77.如实施方案69-76任一项所述的方法，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体、PD-1/PD-L1小分子抑制剂或者TIGIT抗体。
78.如实施方案69-77任一项所述的方法，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体，进一步的，所述抗PD-1/PD-L1抗体为帕博利珠单抗(pembrolizumab)、替雷利珠单抗(Tislelizumab)、尼伏单抗(Nivolumab)、特瑞普利单抗(Toripalimab)、阿替珠单抗(Atezolizumab)、度伐单抗(durvalumab)、阿维单抗(Avelumab)、阿特珠单抗(Atezolizumab)、卡瑞利珠单抗(Camrelizumab)、信迪利单抗(Sintilimab)、西米普利单抗(Cemiplimab)、恩沃利单抗(envafolimab)、BMS-936559、JS003、SHR-1316、GS-4224、AN-4005或者MX-10181。
79.如实施方案69-77任一项所述的用途、药物组合产品或者方法，其中所述免疫检查点抑制剂为PD-1/PD-L1小分子抑制剂，进一步的，所述PD-1/PD-L1小分子抑制剂为INCB-086550、拉泽替尼(Lazertinib)、IMMH-010、CA-170、ABSK043或者RRx-001。
80.如实施方案69-77任一项所述的方法，其中所述免疫检查点抑制剂为TIGIT抗体，进一步的，所述TIGIT抗体为欧司珀利单抗(Ociperlimab/BGB-A1217)、维博利单抗(Vibostolimab)、domvanalimab(AB154)、替瑞利尤单抗(Tiragolumab)、Belrestotug、艾替利单抗(Etigilimab)、ONO-4686、JS-006、AZD-2936、HLX-301、SEA-TGT、M-6223、IBI-939、COM-902、AB-308、AGEN-1777、AK-127、BAT-6021、BAT-6005、ASP-8374、PM-1022、BMS-986207、HB0036或IBI-321。
81.如实施方案69所述的方法，其中所述FAK抑制剂为IN10018或其药学上可接受的盐；所述拓扑异构酶抑制剂为伊立替康(Irinotecan)或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
82.如实施方案69所述的方法，其中所述FAK抑制剂为IN10018或其药学上可接受的盐；所述拓扑异构酶抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)，或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
83.如实施方案69-82任一项所述的方法，所述FAK抑制剂、所述拓扑异构酶抑制剂和所述免疫检查点抑制剂被同时或依次施用于所述对象。
84.如实施方案69-83任一项所述的方法，其中所述肿瘤为膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、神经胶质瘤或者恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)；优选地，所述肿瘤为乳腺癌、卵巢癌、结肠癌(包括结直肠癌)、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤或胰腺癌。
85如实施方案69-84任一项所述的方法，其中所述肿瘤为乳腺癌或结肠癌(包括结直肠癌)。
86.FAK抑制剂在制备用于在对象中治疗肿瘤的药物中的用途，其中将所述FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂施用于所述对象，其中所述拓扑异构酶抑制剂不为蒽环类。
87.拓扑异构酶抑制剂在制备用于在对象中治疗肿瘤的药物中的用途，其中将FAK抑制剂，所述拓扑异构酶抑制剂和免疫检查点抑制剂施用于所述对象，其中所述拓扑异构酶抑制剂不为蒽环类。
88.免疫检查点抑制剂在制备用于在对象中治疗肿瘤的药物中的用途，其中将FAK抑制剂、拓扑异构酶抑制剂和所述免疫检查点抑制剂施用于所述对象，其中所述拓扑异构酶抑制剂不为蒽环类。
89.FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂在制备用于治疗肿瘤的联用药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
90.FAK抑制剂在制备用于与拓扑异构酶抑制剂和免疫检查点抑制剂治疗肿瘤的联用药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
91.拓扑异构酶抑制剂在制备用于与FAK抑制剂和免疫检查点抑制剂治疗肿瘤的联用药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
92.免疫检查点抑制剂在制备用于与FAK抑制剂和拓扑异构酶抑制剂治疗肿瘤的联用药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
93.FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂在制备用于联合治疗肿瘤的药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
94.FAK抑制剂在制备用于与拓扑异构酶抑制剂和免疫检查点抑制剂联合治疗肿瘤的药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
95.拓扑异构酶抑制剂在制备用于与FAK抑制剂和免疫检查点抑制剂联合治疗肿瘤的药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
96.免疫检查点抑制剂在制备用于与FAK抑制剂和拓扑异构酶抑制剂联合治疗肿瘤的药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
97.如实施方案86-96任一项所述的用途，其中所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、APG-2449、VS-4718、AMP945、AMP886或其药学上可接受的盐，优选为IN10018、Defactinib、AMP945或其药学上可接受的盐，进一步优选为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐，所述IN10018结构如下：
98.如实施方案86-97任一项所述的用途，其中所述拓扑异构酶抑制剂为拓扑异构酶I抑制剂。
99.如实施方案98所述的用途，其中所述拓扑异构酶I抑制剂为拓扑替康(Topotecan)、伊立替康(Irinotecan)、贝洛替康(Belotecan)、科西特康(Cositecan)、依喜替康(Exatecan，DX-8951)、茚并异喹啉(Indenoisoquinoline)、Indotecan(LMP-400)、Indimitecan(LMP-776)、希明替康(Simmitecan)、Gimatecan(ST1481)、EC-112002、PLX-038(NK012)、AR-67，或其药学上可接受的盐。
100.如实施方案99所述的用途，其中所述拓扑异构酶I抑制剂为伊立替康(Irinotecan)或其药学上可接受的盐。
101.如实施方案86-97任一项所述的用途，其中所述拓扑异构酶抑制剂为拓扑异构酶II抑制剂。
102.如实施方案101所述的用途，其中所述拓扑异构酶II抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)、莎巴比星(Sabarubicin)，或其药学上可接受的盐。
103.如实施方案102所述的用途，其中所述拓扑异构酶II抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)，或其药学上可接受的盐。
104.如实施方案86-103任一项所述的用途，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体、PD-1/PD-L1小分子抑制剂或者TIGIT抗体。
105.如实施方案86-104任一项所述的用途，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体，进一步的，所述抗PD-1/PD-L1抗体为帕博利珠单抗(pembrolizumab)、替雷利珠单抗(Tislelizumab)、尼伏单抗(Nivolumab)、特瑞普利单抗(Toripalimab)、阿替珠单抗(Atezolizumab)、度伐单抗(durvalumab)、阿维单抗(Avelumab)、阿特珠单抗(Atezolizumab)、卡瑞利珠单抗(Camrelizumab)、信迪利单抗(Sintilimab)、西米普利单抗(Cemiplimab)、恩沃利单抗(envafolimab)、BMS-936559、JS003、SHR-1316、GS-4224、AN-4005或者MX-10181。
106.如实施方案86-104任一项所述的用途、药物组合产品或者方法，其中所述免疫检查点抑制剂为PD-1/PD-L1小分子抑制剂，进一步的，所述PD-1/PD-L1小分子抑制剂为INCB-086550、拉泽替尼(Lazertinib)、IMMH-010、CA-170、ABSK043或者RRx-001。
107.如实施方案86-104任一项所述的用途，其中所述免疫检查点抑制剂为TIGIT抗体，进一步的，所述TIGIT抗体为欧司珀利单抗(Ociperlimab/BGB-A1217)、维博利单抗(Vibostolimab)、domvanalimab(AB154)、替瑞利尤单抗(Tiragolumab)、Belrestotug、艾替利单抗(Etigilimab)、ONO-4686、JS-006、AZD-2936、HLX-301、SEA-TGT、M-6223、IBI-939、COM-902、AB-308、AGEN-1777、AK-127、BAT-6021、BAT-6005、ASP-8374、PM-1022、BMS-986207、HB0036或IBI-321。
108.如实施方案86-96任一项所述的用途，其中所述FAK抑制剂为IN10018或其药学上可接受的盐；所述拓扑异构酶抑制剂为伊立替康(Irinotecan)或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
109.如实施方案86-96任一项所述的用途，其中所述FAK抑制剂为IN10018或其药学上可接受的盐；所述拓扑异构酶抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)，或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
110.如实施方案86-109任一项所述的用途，所述FAK抑制剂、所述拓扑异构酶抑制剂和所述免疫检查点抑制剂被同时或依次施用于所述对象。
111.如实施方案86-110任一项所述的用途，其中所述肿瘤为膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、神经胶质瘤或者恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)；优选地，所述肿瘤为乳腺癌、卵巢癌、结肠癌(包括结直肠癌)、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤或胰腺癌。
112.如实施方案86-111任一项所述的用途，其中所述肿瘤为乳腺癌或结肠癌(包括结直肠癌)。
实施例
提供下面的实施例以进一步阐述本发明。应理解，这些实施例仅用于举例说明本发明，而不用于限制本发明的范围。
下列实施例中未注明具体条件的实验方法均可以按照这类反应的常规条件进行或者按照制造厂商所建议的条件进行。
以下实施例中所用的实验材料和试剂如无特别说明均可从市售渠道获得。
实施例中所用的缩写含义如下：

实施例1：FAK靶点抑制增强乳腺癌细胞针对依托泊苷(Etoposide)免疫原性细胞死亡的研究
实验方案：
1.FAK沉默联合依托泊苷促进乳腺癌细胞MDA-MB-231凋亡
体外实验利用siRNA技术降低FAK的表达水平，与依托泊苷联合使用48小时；利用流式细胞仪，检测对照和FAK沉默的细胞凋亡
2.FAK沉默联合依托泊苷能够有效促进乳腺癌细胞MDA-MB-231产生ICD作用
体外实验利用siRNA技术降低FAK的表达水平，与依托泊苷联合使用48小时；利用流式细胞仪，检测ICD的主要靶标Calreticulin的表达情况。
实验抗体：重组Alexa647荧光Anti-Calreticulin(抗-钙网蛋白)抗体(Abcam，ab196159)，Annexin V细胞凋亡检测试剂盒(Invitrogen,A35110)。
FAK siRNA由吉玛基因提供，序列如下表所示：
F：CCUGUAUGCCUAUCAGCUUTT；
R：AAGCUGAUAGGCAUACAGGTT
实验仪器：
荧光显微镜(Olympus U-HGLGPS)。化学发光成像仪(BIORAD chemidoc touch)
实验结果：
1、FAK沉默后联合依托泊苷显著增加MDA-MB-231细胞凋亡
分别使用终浓度为50nM的对照siRNA或FAK siRNA转染MDA-MB-231细胞。转染24小时后，给予2μM依托泊苷处理48小时，使用Annexin V试剂盒进行细胞染色并使用流式细胞仪进行检测。统计细胞早期和晚期凋亡数值与DMSO对照组进行比较并使用Graphpad 8.0进行绘图，结果显示FAK沉默组与对照组比较，经过依托泊苷处理后，凋亡被显著的增强了，如图1所示。
2、FAK沉默后联合依托泊苷显著上调了ICD靶点Calreticulin释放及暴露
分别使用终浓度为50nM的对照siRNA或FAK siRNA转染MDA-MB-231细胞。转染24小时后，给予2μM依托泊苷处理48小时，用Calreticulin抗体进行荧光染色，流式细胞仪对染色结果经行分析。FlowJo软件结果统计显示：FAK沉默后联合依托泊苷，Calreticulin的释放及暴露均被显著的增强了，如图2所示。
实施例2：FAK靶点抑制增强乳腺癌细胞针对替尼泊苷(Teniposide)免疫原性细胞死亡的研究
实验方案：
1.FAK沉默，联合替尼泊苷促进乳腺癌细胞MDA-MB-231凋亡
体外实验利用siRNA技术降低FAK的表达水平，与teniposide联合使用48小时；利用流式细胞仪，检测对照和FAK沉默的细胞凋亡
2.FAK沉默，联合替尼泊苷能够有效促进乳腺癌细胞MDA-MB-231产生ICD作用
体外实验利用siRNA技术降低FAK的表达水平，与替尼泊苷联合使用48小时；利用流式细胞仪，检测ICD的主要靶标Calreticulin的表达情况。
实验抗体：
重组Alexa647荧光Anti-Calreticulin(抗-钙网蛋白)抗体(Abcam，ab196159)，Annexin V细胞凋亡检测试剂盒(Invitrogen,A35110)。
FAK siRNA由吉玛基因提供，序列如下表所示：
F：CCUGUAUGCCUAUCAGCUUTT；
R：AAGCUGAUAGGCAUACAGGTT
实验仪器：
荧光显微镜(Olympus U-HGLGPS)。化学发光成像仪(BIORAD chemidoc touch)。
实验结果：
1、FAK沉默后联合替尼泊苷显著增加MDA-MB-231细胞凋亡
分别使用终浓度为50nM的对照siRNA或FAK siRNA转染MDA-MB-231细胞。转染24小时后，给予2μM替尼泊苷处理48小时，使用Annexin V试剂盒进行细胞染色并使用流式细胞仪进行检测。统计细胞早期和晚期凋亡数值与DMSO对照组进行比较并使用Graphpad 8.0进行绘图。结果显示FAK沉默组与对照组比较，经过替尼泊苷处理后，凋亡被显著的增强了，如图3所示。
2、FAK沉默后联合替尼泊苷显著上调了ICD靶点Calreticulin释放及暴露
分别使用终浓度为50nM的对照siRNA或FAK siRNA转染MDA-MB-231细胞。转染24小时后，给予2μM替尼泊苷处理48小时，用Calreticulin抗体进行荧光染色，流式细胞仪对染色结果经行分析。FlowJo软件结果统计显示：FAK沉默后联合替尼泊苷，Calreticulin的释放及暴露均被显著的增强了，如图4所示。
实施例3：IN10018和依托泊苷在结肠癌CT26细胞中的协同作用
CT26细胞用RPMI 1640(上海元培，分类号:L210KJ,批号:F210916)+10％FBS(Gibco，分类号:10099-141c,批号:2158737cp)培养，传代两次，当细胞状况良好时，将其放置在96孔板中，3000个细胞/孔。细胞铺展24小时后，加入含有依托泊苷的培养基，设置10个药物作用浓度，第一个浓度为30μM，5倍稀释，最后一个为对照，药物浓度为0，每个药物浓度设置为3个复合孔。同时，用与上述方法相同的药物浓度建立另一组细胞。不同之处在于，每孔加入5μM IN10018，将药物混合，并在37℃下在5％CO2培养箱中培养72小时。
药物作用72小时后，显微镜下观察细胞，向每个孔中加入10μl CCK8检测试剂(Cellorlab，分类号：CX001M，批号：2571100)，在37℃的5％CO2培养箱中培养2-4小时，然后用微孔板读取器OD450(化学发光法)读取孔板。分析结果发现，依托泊苷(来源MCE，分类号：33419-42-0，批号：113741)组的IC50为24.43μM；依托泊苷+5μM IN10018组的IC50为4.86μM。含有IN10018的组的IC50显著小于不含IN10018的组，表明两种药物联合治疗组的疗效优于单药治疗组，如图5所示。
实施例4：IN10018和依托泊苷在结肠癌CT26细胞中的研究
CT26细胞(中科院细胞所)用RPMI 1640(上海元培，分类号:L210KJ,批号:F210916)+10％FBS(Gibco，分类号:10099-141c,批号:2158737cp)培养，传代两次，当细胞状况良好时，将培养液放置在24孔板中。细胞铺展24小时后，设四组，第一组为对照组，加入培养基，第二组为IN10018，浓度为5μM，第三组为依托泊苷(来源MCE，分类号：33419-42-0，批号：113741)，浓度为20μM，第四组为IN10018(5μM)和依托泊苷(20μM)联用组。将药物混合，并在37℃的5％CO2培养箱中培养48小时。
药物作用48小时后，对细胞进行显微镜观察和拍照，并保存照片。然后收集细胞进行流动分析，并用流式缓冲液(PBS+2％FBS)洗涤细胞两次，将0.5μl AF647抗钙网蛋白抗体(abcam，分类号：ab196159，批号：CR33676773)添加到每个孔中，充分混合，在4℃且避光的条件下孵育20分钟。20分钟后，用流式缓冲液(PBS+2％FBS)清洗细胞两次。使用细胞凋亡检测试剂盒(Beyotime，目录号：CL062L，批号：021921210811)，加入195μl膜联蛋白-V-FITC结合物，轻轻混合细胞，然后加入5μl膜联蛋白-V-FITC抗体，轻轻混合。加入10μl PI染料，轻轻混合，室温避光培养15min，然后在流式细胞仪上进行分析。
显微镜下观察细胞，依托泊苷单药组和两药联用组的细胞状态较差，两药联用组最差，细胞死亡较多，而对照组和IN10018组的细胞状态良好。结果显示，双药联用组的CRT阳性率和Annexin V阳性率高于单药组，详见图6和图7。
实施例5：伊立替康(Irinotecan)和IN10018在小鼠结肠癌CT26细胞体外对免疫原性细胞死亡靶点诱导的研究
分组信息见表1。
表1：体外药效实验分组方案
待测化合物信息见表2。
表2：待测化合物信息

实验主要试剂见表3。
表3：实验主要试剂
实验方法与步骤
细胞培养
CT26细胞由应世生物科技(南京)有限公司维持传代。细胞体外单层培养，细胞培养条件为RPMI1640培养基中加10％胎牛血清，置于37℃、5％CO2培养箱中培养。一周两到三次用胰蛋白酶进行常规消化传代处理。当细胞处于指数生长期，贴壁汇合至80％-90％时，收取细胞并进行铺板。
细胞铺板
CT26细胞用胰蛋白酶进行消化后收取细胞并进行计数，根据计数结果，用对应完全培养基对细胞进行稀释，稀释浓度为5万个细胞每毫升，然后进行24孔细胞培养板铺板，每孔铺1ml细胞悬液即5万个细胞。铺板完成后，将细胞放于37℃、5％CO2培养箱中培养。
加入待测化合物
铺板24小时后，在不同孔内分别加入待测化合物伊立替康和IN10018，待测化合物为之前进行统一配制分装，用DMSO进行溶解，分装体积为50μL/支，置于-20℃避光保存，本次实验取出其中一支进行加药及检测。分组及药物浓度如表4所示。
表4：药物分组及加药浓度
收集细胞进行流式检测
细胞用流式缓冲液(DPBS+2％FBS)清洗两遍后，每孔加入0.5μL AF647Anti-Calreticulin Antibody抗体，混匀后4℃避光孵育20min。此后加入流式缓冲液清洗一遍后加入195μL Annexin-V-FITC结合液，轻轻混匀细胞后加入5μL Annexin-V-FITC抗体，进一步混匀后加入10μL PI染料并进行混匀，室温避光孵育15min后进行流式检测。
数据分析
实验结束后，用Flowjo(V10)软件进行细胞阳性率的分析。
实验结果
各组在药物作用后48小时后。流式细胞分析结果显示，联合用药组CRT和Annexin V阳性率均明显高于单药组，详见图8。
实施例6：替尼泊苷(Teniposide)和IN10018对小鼠结肠癌CT26细胞体外增殖抑制活性的研究
实验设计
分组信息见表5。
表5：细胞增殖抑制试验分组方案
待测化合物信息见表6。
表6：
主要试剂信息见表7。
表7：
实验方法与步骤：
细胞培养：
CT26细胞由应世生物科技(南京)有限公司复苏并维持传代。细胞体外单层贴壁培养，培养条件为RPMI1640培养基中加10％胎牛血清，37℃、 5％CO2。一周两到三次使用胰蛋白酶对细胞消化传代。当细胞生长处于指数生长期，贴壁汇合至80％-90％时，收取细胞并进行铺板。
细胞铺板：
CT26细胞用胰蛋白酶进行消化后收取细胞并进行计数，根据计数结果，使用RPMI1640+10％FBS将细胞稀释至每毫升3万个细胞，然后对96孔细胞平底培养板进行细胞铺板，每孔加入0.1ml细胞悬液即3千个细胞。细胞铺板完成后，将培养板置于37℃、5％CO2培养箱中继续培养。
加入待测化合物：
细胞铺板24小时后，在细胞板不同的孔内分别加入待测化合物替尼泊苷，待测化合物在实验前进行统一配制及分装。简言之，使用DMSO将药物溶解至10mM，分装体积为50μL/支，置于-20℃避光保存，本次实验取出其中一支对细胞进行加药及处理。分组及药物浓度信息见表8所示。
表8：药物分组及加药浓度
替尼泊苷设计10个药物作用浓度，其中最后一个药物作用浓度为0，设为阴性对照组。
加药完成后，将药物轻轻混匀，然后细胞置于37℃、5％CO2培养箱中培养。
加入CCK-8检测试剂并读板：
药物作用72小时后，使用多孔道移液器对细胞板每孔均加入10μL CCK-8检测试剂，而后将细胞板置于37℃、5％CO2培养箱中继续孵育4小时。最后使用酶标仪测定每孔于450nm处的吸光度。
数据分析：
实验结束后，使用GraphPad Prism 8软件对测试药物对细胞的抑制百分比进行分析。
抑制百分比计算：
抑制百分比＝{[A(0加药)－A(空白)]--[A(加药)－A(空白)]}/[A(0加
药)－A(空白)]×100％
A(加药)：加有细胞、CCK-8溶液和药物溶液孔的吸光度值
A(空白)：加有培养基和CCK-8溶液而无细胞孔的吸光度值
A(0加药)：加有细胞、CCK-8溶液而无药物溶液孔的吸光度值
实验结论：
本实验评价了替尼泊苷单药和与IN10018合用后在体外条件下对CT26细胞增殖抑制的作用。各组在药物作用后72小时相关检测结果如图9所示。替尼泊苷与IN10018联合用药组的IC50值为0.09799μM，低于单药组的IC50值(0.8179μM)，因此联合用药组具有更强的体外抑制癌细胞生长作用。
实施例7：替尼泊苷和IN10018在小鼠结肠癌CT26细胞体外对免疫原性细胞死亡靶点诱导的研究
实验分组见表9。
表9：
化合物信息见表10：
表10.
化合物信息见表11。
表11：
细胞培养
CT26细胞由应世生物科技(南京)有限公司维持传代。细胞体外单层培养，培养条件为RPMI1640培养基中加10％胎牛血清，37℃、5％CO2培养箱中培养。一周两到三次用胰蛋白酶进行常规消化传代处理。当细胞处于指数生长期，贴壁汇合至80％-90％时，收取细胞并进行铺板。
细胞铺板
CT26细胞用胰蛋白酶进行消化后收取细胞并进行计数，根据计数结果，用RPMI1640+10％FBS对细胞进行稀释，稀释浓度为5万个细胞每毫升，然后进行12孔细胞培养板铺板，每孔铺2ml细胞悬液即10万个细胞。
铺板完成后，将细胞放于37℃、5％CO2培养箱中培养。
加入待测化合物
铺板24小时后，在不同孔内分别加入待测化合物替尼泊苷，待测化合物为之前进行统一配制分装，用DMSO进行溶解，浓度均为10mM，分装体积为50μL/支，置于-20℃避光保存，本次实验取出其中一支进行加药及检测。分组及药物浓度如表9所示。
药物作用48小时后，使用显微镜对细胞进行拍照，然后胰蛋白酶消化细胞并收集细胞进行流式染色。
细胞用流式缓冲液(DPBS+2％FBS)清洗两遍后，每组细胞平均分为两份，其中一份每孔加入0.5μL AF647Anti-Calreticulin Antibody抗体，混匀后4℃避光孵育20分钟。此后加入流式缓冲液清洗一遍后加入195μL Annexin-V-FITC结合液，轻轻混匀细胞后加入5μL Annexin-V-FITC抗体，进一步混匀后加入10μL PI染料并进行混匀，室温避光孵育15分钟后进行流式检测。
数据分析
实验结束后，用Flowjo(V10)软件进行细胞阳性率的分析。
实验结果：
本实验评价了替尼泊苷单药和与IN10018合用后在体外条件下对CT26细胞诱导表达免疫细胞死亡靶点的作用。
各组在药物作用后48小时相关检测结果如图10所示。在显微镜下观察细胞状态，替尼泊苷单药组细胞死亡明显，联合用药组细胞死亡最为显著，阴性对照组和IN10018组细胞活力较好。流式细胞分析结果显示，联合用药组CRT和Annexin V阳性率均明显高于单药组。
实施例8：依托泊苷、IN10018与抗鼠PD-L1抗体之间的配伍治疗在结肠癌MC38细胞C57BL/6小鼠皮下同种移植瘤模型中的体内抗肿瘤药效研究
分组与给药信息见表12。
表12：体内药效实验动物分组及给药方案

注：
1.N：每组小鼠数目；
2.给药体积：根据小鼠体重10mL/kg，如果体重下降超过15％，动物停止给药；待体重恢复至降低10％，
再恢复给药；
3.依托泊苷剂量在分组给药之后的第6天增至6mg/kg给药；
细胞培养：
结直肠癌细胞MC38(来源南京科佰生物科技有限公司，货号：CBP60825)由应世生物科技(南京)有限公司维持传代。细胞体外单层培养，培养条件为DMEM培养基中加10％胎牛血清，37℃，5％CO2培养箱中培养。一周两到三次用胰酶-EDTA进行常规消化处理传代。当细胞处于指数生长期，饱和度为80％-90％时，收取细胞，计数后接种。
细胞接种及分组：
将0.1mL含有2×105个细胞的细胞悬液皮下接种于每只小鼠的右后背。当肿瘤体积达到～50mm3左右时(细胞接种后的第9天)，根据肿瘤体积进行随机分组并给药，受试物及对照溶媒配制见表13。
表13：受试物及对照溶媒配制
实验动物日常观察：
本实验方案的拟定及任何修改均通过了云桥生物IACUC的评估核准。实验动物的使用及福利遵照AAALAC的规定执行。每天监测动物的健康状况及死亡情况，例行检查包括观察肿瘤生长和药物治疗对动物日常行为表现的影响如行为活动，摄食摄水量(仅目测)，体重变化，外观体征或其它不正常情况。基于各组动物数量记录了组内动物死亡数和副作用。
实验终止：
若动物健康状况持续恶化，或瘤体积超过3,000mm3，或有严重疾病，或疼痛，须处以安乐死。有以下情况者，通知兽医并处以安乐死：明显消瘦，体重降低大于20％；不能自由取食和饮水；对照组瘤体积平均值达到3,000mm3，实验终止。动物出现以下临床表现且持续恶化：立毛，弓背，耳、鼻、眼或足色发白，呼吸仓促，抽搐，连续腹泻，脱水，行动迟缓，发声。
肿瘤测量和实验指标：
实验指标是考察肿瘤生长是否被抑制、延缓或治愈。用游标卡尺测量肿瘤直径，每周测量3次。肿瘤体积的计算公式为：V＝0.5×a×b2，a和b分别表示肿瘤的长径和短径。
化合物的抑瘤疗效用TGI(％)评价，反映肿瘤生长抑制率。参照分组后第一天的肿瘤体积，根据以下公式计算肿瘤生长抑制率TGI(％)。TGI(％)＝[1-(某给药组的平均瘤体积-该给药组开始治疗时平均瘤体积)/(溶剂对照组的平均瘤体积-溶剂对照组开始治疗时平均瘤体积)]×100％。
统计分析：
统计分析基于试验结束时肿瘤体积运用Prism Graphpad软件进行分析。多组间比较，采用Two-way ANOVA，Fisher’s LSD test法检验进行分析。P<0.05认为有显著性差异。
实验结果：
1.受试物依托泊苷、IN10018与抗鼠PD-L1抗体之间的配伍治疗在MC38小鼠结直肠癌细胞C57BL/6小鼠皮下同种移植瘤模型中的体内药效研究
细胞接种后，每天观察肿瘤生长情况，接种后第9天根据肿瘤体积进行分组，入组平均肿瘤体积约为50mm3。由于肿瘤负荷，对照组在接种后的第26天，即分组给药之后的第17天安乐死，整个实验结束。
分组给药后第17天，对照组的肿瘤体积为2670.9±1438.9mm3。PD-L1抗体(5mg/kg)和依托泊苷(3mg/kg)单药治疗组的肿瘤体积分别为2677.7±2029.0mm3和2420.2±1377.3mm3；依托泊苷+PD-L1抗体(3+5mg/kg)和依托泊苷+IN10018(3+25mg/kg)两药联用治疗组的肿瘤体积分别为1947.6±539.7mm3和1865.0±839.9mm3；依托泊苷+IN10018+PD-L1抗体(3+25+5mg/kg)三药联用治疗组的肿瘤体积分别为1245.6±1064.7mm3。综合肿瘤体积与对照组进行比较，PD-L1抗体(5mg/kg)和依托泊苷(3mg/kg)单药治疗组的抑瘤率TGI分别为-0.2％(p＝0.9827)和9.5％(p＝0.4487)；依托泊苷+PD-L1抗体(3+5mg/kg)和依托泊苷+IN10018(3+25mg/kg)两药联用治疗组的抑瘤率TGI分别为27.6％(p＝0.0295)和30.7％(p＝0.0111)；依托泊苷+IN10018+PD-L1抗体(3+25+5mg/kg)三药联用治疗组的抑瘤率TGI为54.4％(p<0.0001)。
综合瘤体积与依托泊苷+IN10018+PD-L1抗体(3+25+5mg/kg)三药联用组相比较，进行统计学分析，对照组、PD-L1抗体(5mg/kg)和依托泊苷(3mg/kg)单药治疗组的P值分别为p<0.0001、p<0.0001和p＝0.0005；依托泊苷+PD-L1抗体(3+5mg/kg)和依托泊苷+IN10018(3+25mg/kg)两药联用治疗组的P值分别为p＝0.0346和p＝0.0504，详见表14。各剂量组在不同时间段的肿瘤体积如图11所示。与空白对照组相比，PD-L1抗体(5mg/kg)和依托泊苷(3mg/kg)单药治疗组的平均瘤体积和对照组的平均瘤体积很接近，并没有显现出对肿瘤生长有抑制作用；而依托泊苷+PD-L1抗体(3+5mg/kg)和依托泊苷+IN10018(3+25mg/kg)两药联用治疗组和依托泊苷+IN10018+PD-L1抗体(3+25+5mg/kg)三药联用组则均有明显肿瘤生长抑制作用，和对照组相比都有统计学差异。综合整个给药周期，依托泊苷+IN10018+PD-L1抗体(3+25+5mg/kg)三药联用组相对于其余各治疗组的肿瘤体积一直都更小，且和PD-L1抗体(5mg/kg)和依托泊苷(3mg/kg)单药治疗组以及依托泊苷+PD-L1抗体(3+5mg/kg)两药联用治疗组相比均具有统计学差异，虽然依托泊苷+IN10018+PD-L1抗体(3+25+5mg/kg)三药联用组和依托泊苷+IN10018(3+25mg/kg)两药联用治疗组相比没有统计学差异，但是其平均瘤体积相对而言一直都较小，也显示出依托泊苷+IN10018+PD-L1抗体(3+25+5mg/kg)三药联用组具有更好的抑制肿瘤生长的效果。
表14：受试物对小鼠结肠癌MC38细胞的C57BL/6小鼠移植瘤模型的抑瘤效果评价(基于分组给药后第17天数据)

注：
1.按照分组给药后的天数来计算，数据为平均值±标准差(mean±SD)；
2.TGI(％)＝[1-(T17-T0)/(V17-V0)]×100％；
3.*：p<0.05，vs.对照组，Two-way ANOVA；
4.*：p<0.05，vs.依托泊苷+IN10018+PD-L1抗体(3+25+5mg/kg)组，Two-way ANOVA；
2.受试物依托泊苷、IN10018和抗鼠PD-L1抗体之间的配伍治疗在MC38小鼠结直肠癌细胞C57BL/6小鼠皮下同种移植瘤模型中的体重变化及临床状况研究
实验按照给药方案进行，实验过程中，每天观察动物摄食饮水等活动，每周记录3次动物体重。分组给药17天之后，对照组的平均体重从分组给药当天(Day0)的18.7g，变化至23.9g，体重增长率为28.8％；PD-L1抗体(5mg/kg)和依托泊苷(3mg/kg)单药治疗组的平均体重分别从Day0的18.8g和19.1g变化至Day17的23.6g和22.8g，其体重变化率分别为25.8％和19.3％；依托泊苷+PD-L1抗体(3+5mg/kg)和依托泊苷+IN10018(3+25mg/kg)两药联用治疗组的平均体重分别从Day0的19.1g和18.9g变化至Day17的22.3g和22.5g，体重变化率分别为17.2％和19.0％；依托泊苷+IN10018+PD-L1抗体(3+25+5mg/kg)三药联用组的平均体重从Day 0的19.1g变化至Day17的21.6g，体重变化率为13.3％。在整个给药周期中，依托泊苷(3mg/kg)单药治疗组和依托泊苷+PD-L1抗体(3+5mg/kg)两药联用治疗组分别各有1只小鼠，由于肿瘤破溃，分别在分组给药后的第12天和第17天发现死亡，其余各组的动物在整个给药周期中，体重均无明显下降且状态良好，显示出对不同给药方式的耐受。详见表15。各剂量组在不同时间段的体重变化如图12所示。说明动物对依托泊苷+IN10018+PD-L1抗体(3+25+5mg/kg)三药联用耐受。
表15：受试物对小鼠结肠癌MC38细胞的C57BL/6小鼠移植瘤模型的体重变化评价(基于分组给药后第17天数据)

注：
1.按照分组给药后的天数来计算，第0天的动物存活数目/第17天的动物存活数目；
2.数据为平均值±标准差(mean±SD)；
3.体重变化率＝[1-(W17-W0)/W0]*100％；
实施例9：依托泊苷(Etoposide)和AMP945对小鼠乳腺癌4T1细胞体外免疫原性细胞死亡靶点诱导的研究
实验材料：
1)本实验所用药物
Etoposide由MCE提供,Lot No.:113741
AMP945由MCE提供,Lot No.:143253
2)本实验所用抗体
重组Alexa647荧光Anti-Calreticulin抗体(Abcam，Cat No.:ab196159,Lot No.:CR33676773)。Annexin V-调亡检测试剂盒(Beyotime,Cat No.:C1062L,Lot No.:122221220706)。
实验方法：
4T1细胞(来源南京科佰生物科技有限公司，货号：CBP60352)用RPMI1640(上海元培,Cat No.:L210KJ,Lot No.:F210916)+10％FBS(Gibco,Cat No.:10099-141c,Lot No.:2158737cp)培养，培养条件为37℃、5％CO2。一周两到三次用胰蛋白酶进行常规消化传代处理。当细胞处于指数生长期，贴壁汇合至80％-90％时，收取细胞并进行铺板。4T1细胞用胰蛋白酶进行消化后收取细胞并进行计数，根据计数结果，用RPMI1640+10％FBS对细胞进行稀释，稀释浓度为5万个细胞每毫升，然后进行12孔细胞培养板铺板，每孔铺2ml细胞悬液即10万个细胞。铺板完成后，将细胞放于37℃、5％CO2培养箱中培养。细胞铺展24小时后，设立六组，第一组为对照组，添加培养基，第二组为AMP945，浓度为3μM，第三组为AMP945，浓度为6μM，第四组为Etoposide，浓度为5μM，第五组为AMP945(3μM)与Etoposide(5μM)联用，第六组为AMP945(6μM)与Etoposide(5μM)联用。将药物混合，并在37℃下在5％CO2培养箱中培养48小时。
实验结果
药物作用48h后，收集细胞进行流动分析，并使用流式缓冲液(PBS+2％FBS)清洗细胞两次，每个孔中加入0.5μl AF647抗钙网蛋白抗体(abcam)，混合。在4℃且避光的条件下孵育，孵育20min后加入流式缓冲液，使用Annexin V-调亡检测试剂盒(Beyotime)，加入195μl Annexin-V-FITC结合物，与细胞进行吹打混合后，加入5μl Annexin-V-FITC抗体，轻轻混合，最后加入10μl PI染料混合，室温避光孵育15min，将样品送至流式细胞仪上进行信号测定。
流式细胞仪分析结果显示两药合用组的CRT阳性率和Annexin-V阳性率显著优于单药组及对照组，见图13。
实施例10：伊立替康(Irinotecan)和AMP945对小鼠乳腺癌4T1细胞体外免疫原性细胞死亡靶点诱导的研究
实验材料：
1)本实验所用药物
Irinotecan由上海超岚化工科技中心提供,Lot No.:202110
AMP945由MCE提供,Lot No.:143253
2)本实验所用抗体
重组Alexa647荧光Anti-Calreticulin抗体(Abcam，Cat No.:ab196159,Lot No.:CR33676773)。Annexin V-调亡检测试剂盒(Beyotime,Cat No.:C1062L,Lot No.:122221220706)。
实验方法：
4T1细胞(来源南京科佰生物科技有限公司，货号：CBP60352)用RPMI1640(上海元培,Cat No.:L210KJ,Lot No.:F210916)+10％FBS(Gibco,Cat No.:10099-141c,Lot No.:2158737cp)培养，培养条件为37℃、5％CO2。一周两到三次用胰蛋白酶进行常规消化传代处理。当细胞处于指数生长期，贴壁汇合至80％-90％时，收取细胞并进行铺板。4T1细胞用胰蛋白酶进行消化后收取细胞并进行计数，根据计数结果，用RPMI1640+10％FBS对细胞进行稀释，稀释浓度为5万个细胞每毫升，然后进行12孔细胞培养板铺板，每孔铺2ml细胞悬液即10万个细胞。铺板完成后，将细胞放于37℃、5％CO2培养箱中培养。细胞铺展24小时后，设立六组，第一组为对照组，添加培养基，第二组为AMP945，浓度为3μM，第三组为AMP945，浓度为6μM，第四组为Irinotecan浓度为120μM，第五组为AMP945(3μM)与Irinotecan(120μM)联用，第六组为AMP945(6μM)与Irinotecan(120μM)联用。将药物混合，并在37℃下在5％CO2培养箱中培养48小时。
实验结果
药物作用48h后，收集细胞进行流动分析，并使用流式缓冲液(PBS+2％FBS)清洗细胞两次，每个孔中加入0.5μl AF647抗钙网蛋白抗体(abcam)，混合。在4℃且避光的条件下孵育，孵育20min后加入流式缓冲液，使用Annexin V-调亡检测试剂盒(Beyotime)，加入195μl Annexin-V-FITC结合物，与细胞进行吹打混合后，加入5μl Annexin-V-FITC抗体，轻轻混合，最后加入10μl PI染料混合，室温避光孵育15min，将样品送至流式细胞仪上进行信号测定。
流式细胞仪分析结果显示两药合用组的CRT阳性率和Annexin-V阳性率显著优于单药组及对照组，见图14。
通过引用将本发明中所提及的所有参考文献均完整合并入本文，就如同每一篇文献均单独列出一样。应理解，在阅读了本发明的公开内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落入本申请所附权利要求书所限定的范围内。

Claims (48)

  1. FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂在制备用于在对象中治疗肿瘤的药物中的用途，其中所述拓扑异构酶抑制剂不为蒽环类。
  2. FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂的药物组合产品，其用于在对象中治疗肿瘤，其中所述拓扑异构酶抑制剂不为蒽环类。
  3. 一种治疗肿瘤的方法，该方法包括向对象施用治疗有效量的FAK抑制剂、拓扑异构酶抑制剂和免疫检查点抑制剂，其中所述拓扑异构酶抑制剂不为蒽环类。
  4. 如权利要求1-3任一项所述的用途、药物组合产品或者方法，其中所述FAK抑制剂和所述拓扑异构酶抑制剂诱导免疫原性细胞死亡(ICD)，其中所述拓扑异构酶抑制剂不为蒽环类。
  5. 如权利要求1-4任一项所述的用途、药物组合产品或者方法，其中所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、APG-2449、AMP945、AMP886或其药学上可接受的盐，优选为IN10018、Defactinib、AMP945或其药学上可接受的盐，进一步优选为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐，所述IN10018结构如下：
  6. 如权利要求1-5任一项所述的用途、药物组合产品或者方法，其中所述拓扑异构酶抑制剂为拓扑异构酶I抑制剂。
  7. 如权利要求6所述的用途、药物组合产品或者方法，其中所述拓扑异构酶I抑制剂为拓扑替康(Topotecan)、伊立替康(Irinotecan)、贝洛替康(Belotecan)、科西特康(Cositecan)、依喜替康(Exatecan，DX-8951)、茚并异喹啉(Indenoisoquinoline)、Indotecan(LMP-400)、Indimitecan(LMP-776)、希明替康(Simmitecan)、Gimatecan(ST1481)、EC-112002、PLX-038(NK012)、AR-67，或其药学上可接受的盐。
  8. 如权利要求7所述的用途、药物组合产品或者方法，其中所述拓扑异构酶I抑制剂为伊立替康(Irinotecan)，或其药学上可接受的盐。
  9. 如权利要求1-5任一项所述的用途、药物组合产品或者方法，其中所述拓扑异构酶抑制剂为拓扑异构酶II抑制剂。
  10. 如权利要求9所述的用途、药物组合产品或者方法，其中所述拓扑异构酶II抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)、莎巴比星(Sabarubicin)，或其药学上可接受的盐。
  11. 如权利要求10所述的用途、药物组合产品或者方法，其中所述拓扑异构酶II抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)，或其药学上可接受的盐。
  12. 如权利要求1-11任一项所述的用途、药物组合产品或者方法，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体、PD-1/PD-L1小分子抑制剂或者TIGIT抗体。
  13. 如权利要求1-12任一项所述的用途、药物组合产品或者方法，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体，进一步的，所述抗PD-1/PD-L1抗体为帕博利珠单抗(pembrolizumab)、替雷利珠单抗(Tislelizumab)、尼伏单抗(Nivolumab)、特瑞普利单抗(Toripalimab)、阿替珠单抗(Atezolizumab)、度伐单抗(durvalumab)、阿维单抗(Avelumab)、阿特珠单抗(Atezolizumab)、卡瑞利珠单抗(Camrelizumab)、信迪利单抗(Sintilimab)、西米普利单抗(Cemiplimab)、恩沃利单抗(envafolimab)、BMS-936559、JS003、SHR-1316、GS-4224、AN-4005或者MX-10181。
  14. 如权利要求1-12任一项所述的用途、药物组合产品或者方法，其中所述免疫检查点抑制剂为PD-1/PD-L1小分子抑制剂，进一步的，所述PD-1/PD-L1小分子抑制剂为INCB-086550、拉泽替尼(Lazertinib)、IMMH-010、CA-170、ABSK043或者RRx-001。
  15. 如权利要求1-12任一项所述的用途、药物组合产品或者方法，其中所述免疫检查点抑制剂为TIGIT抗体，进一步的，所述TIGIT抗体为欧司珀利单抗(Ociperlimab/BGB-A1217)、维博利单抗(Vibostolimab)、domvanalimab(AB154)、替瑞利尤单抗(Tiragolumab)、Belrestotug、艾替利单抗(Etigilimab)、ONO-4686、JS-006、AZD-2936、HLX-301、SEA-TGT、M-6223、IBI-939、COM-902、AB-308、AGEN-1777、AK-127、BAT-6021、BAT-6005、ASP-8374、PM-1022、BMS-986207、HB0036或IBI-321。
  16. 如权利要求1-5任一项所述的用途、药物组合产品或者方法，其中所述FAK抑制剂为IN10018或其药学上可接受的盐；所述拓扑异构酶抑制剂为伊立替康(Irinotecan)或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
  17. 如权利要求1-5任一项所述的用途、药物组合产品或者方法，其中所述FAK抑制剂为IN10018或其药学上可接受的盐；所述拓扑异构酶抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)，或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
  18. 如权利要求1-17任一项所述的用途、药物组合产品或者方法，所述FAK抑制剂、所述拓扑异构酶抑制剂和所述免疫检查点抑制剂被同时或依次施用于所述对象。
  19. 如权利要求1-18任一项所述的用途、药物组合产品或者方法，其中所述肿瘤为膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、神经胶质瘤或者恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)；优选地，所述肿瘤为乳腺癌、卵巢癌、结肠癌(包括结直肠癌)、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤或胰腺癌。
  20. 如权利要求1-19任一项所述的用途、药物组合产品或者方法，其中所述肿瘤为乳腺癌或结肠癌(包括结直肠癌)。
  21. 一种试剂盒或药学上可接受的组合物，其包括：
    (a)FAK抑制剂；
    (b)拓扑异构酶抑制剂，所述拓扑异构酶抑制剂不为蒽环类；和
    (c)免疫检查点抑制剂。
  22. 如权利要求21所述的试剂盒或组合物，其中所述FAK抑制剂和所述拓扑异构酶抑制剂诱导免疫原性细胞死亡(ICD)。
  23. 如权利要求21-22任一项所述的试剂盒或组合物，其中所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、APG-2449、AMP945、AMP886或其药学上可接受的盐，优选为IN10018、Defactinib、AMP945或其药学上可接受的盐，进一步优选为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐，所述IN10018结构如下：
  24. 如权利要求21-23任一项所述的试剂盒或组合物，其中所述拓扑异构酶抑制剂为拓扑异构酶I抑制剂。
  25. 如权利要求24所述的试剂盒或组合物，其中所述拓扑异构酶I抑制剂为拓扑替康(Topotecan)、伊立替康(Irinotecan)、贝洛替康(Belotecan)、科西特康(Cositecan)、依喜替康(Exatecan，DX-8951)、茚并异喹啉(Indenoisoquinoline)、Indotecan(LMP-400)、Indimitecan(LMP-776)、希明替康(Simmitecan)、Gimatecan(ST1481)、EC-112002、PLX-038(NK012)、AR-67，或其药学上可接受的盐。
  26. 如权利要求25所述的试剂盒或组合物，其中所述拓扑异构酶I抑制剂为伊立替康(Irinotecan)，或其药学上可接受的盐。
  27. 如权利要求21-23任一项所述的试剂盒或组合物，其中所述拓扑异构酶抑制剂为拓扑异构酶II抑制剂。
  28. 如权利要求27所述的试剂盒或组合物，其中所述拓扑异构酶II抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)、莎巴比星(Sabarubicin)，或其药学上可接受的盐。
  29. 如权利要求28所述的试剂盒或组合物，其中所述拓扑异构酶II抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)，或其药学上可接受的盐。
  30. 如权利要求21-29任一项所述的试剂盒或组合物，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体、PD-1/PD-L1小分子抑制剂或者TIGIT抗体。
  31. 如权利要求21-30任一项所述的试剂盒或组合物，其中所述免疫检查点抑制剂为抗PD-1/PD-L1抗体，进一步的，所述抗PD-1/PD-L1抗体为帕博利珠单抗(pembrolizumab)、替雷利珠单抗(Tislelizumab)、尼伏单抗(Nivolumab)、特瑞普利单抗(Toripalimab)、阿替珠单抗(Atezolizumab)、度伐单抗(durvalumab)、阿维单抗(Avelumab)、阿特珠单抗(Atezolizumab)、卡瑞利珠单抗(Camrelizumab)、信迪利单抗(Sintilimab)、西米普利单抗(Cemiplimab)、恩沃利单抗(envafolimab)、BMS-936559、JS003、SHR-1316、GS-4224、AN-4005或者MX-10181。
  32. 如权利要求21-30任一项所述的试剂盒或组合物，其中所述免疫检查点抑制剂为PD-1/PD-L1小分子抑制剂，进一步的，所述PD-1/PD-L1小分子抑制剂为INCB-086550、拉泽替尼(Lazertinib)、IMMH-010、CA-170、ABSK043或者RRx-001。
  33. 如权利要求21-30任一项所述的试剂盒或组合物，其中所述免疫检查点抑制剂为TIGIT抗体，进一步的，所述TIGIT抗体为欧司珀利单抗(Ociperlimab/BGB-A1217)、维博利单抗(Vibostolimab)、domvanalimab(AB154)、替瑞利尤单抗(Tiragolumab)、Belrestotug、艾替利单抗(Etigilimab)、ONO-4686、JS-006、AZD-2936、HLX-301、SEA-TGT、M-6223、IBI-939、COM-902、AB-308、AGEN-1777、AK-127、BAT-6021、BAT-6005、ASP-8374、PM-1022、BMS-986207、HB0036或IBI-321。
  34. 如权利要求21-22任一项所述的试剂盒或组合物，其中所述FAK抑制剂为IN10018或其药学上可接受的盐；所述拓扑异构酶抑制剂为伊立替康(Irinotecan)或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
  35. 如权利要求21-22任一项所述的试剂盒或组合物，其中所述FAK抑制剂为IN10018或其药学上可接受的盐；所述拓扑异构酶抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)，或其药学上可接受的盐；所述免疫检查点抑制剂为抗PD-1/PD-L1抗体或PD-1/PD-L1小分子抑制剂，尤其是抗PD-1/PD-L1抗体。
  36. 如权利要求21-35任一项所述的试剂盒或组合物，其用作药物。
  37. 如权利要求21-36任一项所述的试剂盒或组合物，其中所述药物用于治疗肿瘤，所述肿瘤为膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、神经胶质瘤或者恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)；优选地，所述肿瘤为乳腺癌、卵巢癌、结肠癌(包括结直肠癌)、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤或胰腺癌。
  38. 如权利要求37所述的试剂盒或组合物，其中所述肿瘤为乳腺癌或结肠癌(包括结直肠癌)。
  39. FAK抑制剂，其在治疗肿瘤中用于增强拓扑异构酶抑制剂诱导的免疫原性细胞死亡，其中所述拓扑异构酶抑制剂不为蒽环类。
  40. 如权利要求39所述的FAK抑制剂，其中所述FAK抑制剂为IN10018、Defactinib、GSK2256098、PF-00562271、VS-4718、APG-2449、AMP945、AMP886或其药学上可接受的盐，优选为IN10018、Defactinib、AMP945或其药学上可接受的盐，进一步优选为IN10018或其药学上可接受的盐，尤其是IN10018酒石酸盐，所述IN10018结构如下：
  41. 如权利要求39或40所述的FAK抑制剂，其中所述拓扑异构酶抑制剂为拓扑异构酶I抑制剂。
  42. 如权利要求41所述的FAK抑制剂，其中所述拓扑异构酶I抑制剂为拓扑替康(Topotecan)、伊立替康(Irinotecan)、贝洛替康(Belotecan)、科西特康(Cositecan)、依喜替康(Exatecan，DX-8951)、茚并异喹啉(Indenoisoquinoline)、Indotecan(LMP-400)、Indimitecan(LMP-776)、希明替康(Simmitecan)、Gimatecan(ST1481)、EC-112002、PLX-038(NK012)、AR-67，或其药学上可接受的盐。
  43. 如权利要求42所述的FAK抑制剂，其中所述拓扑异构酶I抑制剂为伊立替康(Irinotecan)，或其药学上可接受的盐。
  44. 如权利要求39或40所述的FAK抑制剂，其中所述拓扑异构酶抑制剂为拓扑异构酶II抑制剂。
  45. 如权利要求44所述的FAK抑制剂，其中所述拓扑异构酶II抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)、莎巴比星(Sabarubicin)，或其药学上可接受的盐。
  46. 如权利要求45所述的FAK抑制剂，其中所述拓扑异构酶II抑制剂为依托泊苷(Etoposide)、替尼泊苷(Teniposide)，或其药学上可接受的盐。
  47. 如权利要求39-46任意一项所述的FAK抑制剂，其中所述肿瘤为膀胱癌、乳腺癌、子宫颈癌、结肠癌(包括结直肠癌)、食管癌、食管鳞状细胞癌、头颈癌、肝癌、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤、骨髓瘤、横纹肌肉瘤、炎性肌纤维母细胞瘤、成神经细胞瘤、胰腺癌、前列腺癌、肾癌、肾细胞癌、肉瘤(包括骨肉瘤)、皮肤癌(包括鳞状细胞癌)、胃癌、睾丸癌、甲状腺癌、子宫癌、间皮瘤、胆管癌、平滑肌肉瘤、脂肪肉瘤、鼻咽癌、神经内分泌癌、卵巢癌、唾液腺癌、梭形细胞癌引起的转移瘤、间变性大细胞淋巴瘤、甲状腺未分化癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、神经胶质瘤或者恶性血液病，例如急性髓细胞性白血病(AML)、急性淋巴细胞白血病(ALL)、弥漫性大B细胞淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)；优选地，所述肿瘤为乳腺癌、卵巢癌、结肠癌(包括结直肠癌)、肺癌(包括小细胞肺癌和非小细胞肺癌)、黑色素瘤或胰腺癌。
  48. 如权利要求39-47任意一项所述的FAK抑制剂，其中所述肿瘤为乳腺癌或结肠癌(包括结直肠癌)。
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CN108289892A (zh) * 2015-06-29 2018-07-17 维瑞斯特姆股份有限公司 治疗组合物、组合和使用方法
CN111867589A (zh) * 2018-03-14 2020-10-30 默克专利股份有限公司 在对象中治疗肿瘤的化合物及其用途

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Publication number Priority date Publication date Assignee Title
CN108289892A (zh) * 2015-06-29 2018-07-17 维瑞斯特姆股份有限公司 治疗组合物、组合和使用方法
CN111867589A (zh) * 2018-03-14 2020-10-30 默克专利股份有限公司 在对象中治疗肿瘤的化合物及其用途

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