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WO2022042537A1 - TGF-β受体的融合蛋白与多靶点酪氨酸激酶抑制剂联合在制备抗肿瘤药物中的用途 - Google Patents

TGF-β受体的融合蛋白与多靶点酪氨酸激酶抑制剂联合在制备抗肿瘤药物中的用途 Download PDF

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Publication number
WO2022042537A1
WO2022042537A1 PCT/CN2021/114280 CN2021114280W WO2022042537A1 WO 2022042537 A1 WO2022042537 A1 WO 2022042537A1 CN 2021114280 W CN2021114280 W CN 2021114280W WO 2022042537 A1 WO2022042537 A1 WO 2022042537A1
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WIPO (PCT)
Prior art keywords
once
tgf
weeks
seq
fusion protein
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PCT/CN2021/114280
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English (en)
French (fr)
Inventor
张晓静
孟志强
解婧
李华军
王莹
Original Assignee
江苏恒瑞医药股份有限公司
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Publication of WO2022042537A1 publication Critical patent/WO2022042537A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/475—Growth factors; Growth regulators
    • C07K14/495—Transforming growth factor [TGF]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
    • 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 present disclosure belongs to the field of pharmacy, in particular to the use of a fusion protein containing a TGF- ⁇ receptor in combination with a multi-target tyrosine kinase inhibitor in the preparation of a medicament for treating tumors.
  • PD-1 Programmed death 1
  • PD-1 is a member of the CD28 superfamily. PD-1 is expressed in activated T cells, B cells and myeloid cells, and it has two ligands, programmed death ligand 1 (PD-L1) and PD-L2. PD-L1 interacts with the receptor PD-1 on T cells and plays an important role in the negative regulation of immune responses.
  • the expression of PD-L1 protein can be detected in many human tumor tissues.
  • the microenvironment of the tumor site can induce the expression of PD-L1 on tumor cells.
  • the expressed PD-L1 is beneficial to the occurrence and growth of tumors and induces anti-tumor effects. Apoptosis of T cells.
  • PD-1/PD-L1 pathway inhibitors can block the combination of PD-1 and PD-L1, block negative regulatory signals, and restore the activity of T cells, thereby enhancing the immune response. Therefore, PD-1/PD- L1-targeted immune regulation has important implications for tumor suppression.
  • TGF- ⁇ Transforming growth factor- ⁇ belongs to the TGF- ⁇ superfamily that regulates cell growth and differentiation.
  • TGF-beta signals through a heterotetrameric receptor complex consisting of two type I and two type II transmembrane serine/threonine kinase receptors.
  • TGF- ⁇ signaling pathway can reduce tumor metastasis.
  • truncated Smad2/3 dominant-negative mutants to inhibit the TGF- ⁇ signaling pathway in breast tumor cell lines, it was found that the metastatic ability of tumor cells was inhibited.
  • Microsatellite instability studies in colon cancer found that inactive mutations in TGF- ⁇ RII resulted in reduced metastasis and increased postoperative survival in subjects in need.
  • the inhibitor that inhibits the TGF- ⁇ signaling pathway alone has a weak effect in clinical treatment, which may be related to the abnormally high expression of TGF- ⁇ in tumor cells, while the inhibitor of the TGF- ⁇ signaling pathway alone is very effective. It is difficult to focus on tumor, resulting in low efficacy or bioavailability of signaling pathway inhibitors.
  • inhibiting the PD-1/PD-L1 pathway on the basis of targeting and neutralizing TGF- ⁇ in the tumor microenvironment can restore the activity of T cells, enhance the immune response, and more effectively improve the effect of inhibiting the occurrence and development of tumors.
  • the PD-L1/TGF- ⁇ dual antibody M-7824 developed by Merck has entered clinical phase III.
  • this product has also been developed in combination with eribulin mesylate for the treatment of metastasis triple-negative breast cancer (NCT03579472); combined with gemcitabine for the treatment of previously treated advanced pancreatic adenocarcinoma (NCT03451773); combined with topotecan or temozolomide for the treatment of recurrent small cell lung cancer (NCT0-3554473) and other indications and, M7824 in NSCLC (NCT03631706) has been terminated early, and a Phase II/III clinical trial of M7824 in combination with gemcitabine and cisplatin in biliary tract cancer (NCT04066491) is currently recruiting.
  • WO2018205985A discloses a novel PD-L1/TGF- ⁇ fusion protein.
  • CN101007815A provides multi-target tyrosine kinase inhibitors, including compounds of formula (I), and discloses that the compounds have VEGF and EGFR inhibitory activities.
  • CN106535896A discloses that compounds of formula (I) are useful for the treatment of fibrotic diseases, such as pulmonary fibrosis, liver cirrhosis, scleroderma or renal fibrosis. The contents of the aforementioned patents are incorporated in this disclosure in their entirety.
  • Pancreatic cancer and biliary tract malignant tumors have a high degree of malignancy and rapid progression, but their onset is insidious and their early symptoms are not typical.
  • the treatment of pancreatic cancer and biliary tract malignancies is a systematic project, which often requires a multidisciplinary consultation model (MDT), which mainly includes surgical treatment, radiotherapy, chemotherapy, interventional therapy and supportive care, in which surgical resection is pancreatic cancer and biliary tract.
  • MDT multidisciplinary consultation model
  • Subjects in need of malignancy have the only chance for cure and long-term survival. For advanced pancreatic cancer that has failed first-line treatment, the available treatment options are limited.
  • gemcitabine or fluorouracil-based single-agent chemotherapy or combination chemotherapy and second-line chemotherapy can be selected.
  • ORR ⁇ 10%.
  • the preferred chemotherapy regimen is oxaliplatin combined with 5-FU
  • immunotherapy regimens include pembrolizumab combined with lenvatinib, pembrolizumab ( Only tumors with high microsatellite instability (MSI-H)/mismatch repair deficiency (dMMR) and nivolumab have been recommended by the Chinese Society of Clinical Oncology (CSCO) and the National Comprehensive Cancer Network (NCCN) guidelines.
  • CSCO Chinese Society of Clinical Oncology
  • NCCN National Comprehensive Cancer Network
  • the present disclosure relates to the use of a fusion protein containing a TGF- ⁇ receptor in combination with a multi-target tyrosine kinase inhibitor in the preparation of a medicament for treating tumors or cancer.
  • the present disclosure provides a method of treating, alleviating or preventing a tumor or cancer, comprising administering to a subject in need thereof a therapeutically, alleviating or prophylactically effective amount of a multi-targeted tyrosine kinase inhibitor and a therapeutically, alleviating or prophylactically effective amount
  • a method of treating, alleviating or preventing a tumor or cancer comprising administering to a subject in need thereof a therapeutically, alleviating or prophylactically effective amount of a multi-targeted tyrosine kinase inhibitor and a therapeutically, alleviating or prophylactically effective amount
  • the fusion protein containing TGF- ⁇ receptor comprising administering to a subject in need thereof a therapeutically, alleviating or prophylactically effective amount of a multi-targeted tyrosine kinase inhibitor and a therapeutically, alleviating or prophylactically effective amount
  • the fusion protein containing TGF- ⁇ receptor comprising administering to a subject in need thereof
  • a method of treating a tumor or cancer comprising administering to a subject in need thereof a therapeutically, ameliorating or prophylactically effective amount of the above-described multi-targeted tyrosine kinase inhibitor, said multi-targeted tyrosine kinase inhibitor
  • the agent is used in combination with the fusion protein containing the TGF- ⁇ receptor.
  • a method of treating a tumor or cancer comprising administering to a subject in need thereof a therapeutically, ameliorating or prophylactically effective amount of a TGF-beta receptor-containing fusion protein, the TGF-beta receptor-containing fusion protein In combination with the multi-targeted tyrosine kinase inhibitor.
  • the multi-target tyrosine kinase inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof,
  • the multi-target tyrosine kinase inhibitor is famitinib or a pharmaceutically acceptable salt thereof.
  • the fusion protein of TGF- ⁇ receptor comprises PD-L1 antibody or antigen-binding fragment thereof and TGF- ⁇ receptor, wherein the part of TGF- ⁇ receptor is the extracellular domain of TGF- ⁇ RII N-terminal truncated form.
  • the TGF- ⁇ receptor fusion protein is shown in general formula (II):
  • TGF- ⁇ RII ECD is a truncated form of the extracellular domain of TGF- ⁇ RII
  • Ab is a PD-L1 antibody or an antigen-binding fragment thereof
  • L is the linking sequence
  • the linker sequence is (G4S)xG, wherein x is 3-6, eg, 4 or 5.
  • the PD-L1 antibody or antigen-binding fragment thereof comprises:
  • HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 respectively, and HCDR1 as shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6 respectively of LCDR1, LCDR2 and LCDR3,
  • HCDR1 SYWMH SEQ ID NO: 1
  • HCDR3 GGSSYDYFDY SEQ ID NO:3
  • LCDR1 RASESVSIHGTHLMH SEQ ID NO:4
  • the PD-L1 antibody or antigen-binding fragment thereof is referred to as a chimeric antibody or a functional fragment thereof, a humanized antibody or a functional fragment thereof, or a human antibody or a functional fragment thereof.
  • sequences of the humanized PD-L1 antibody heavy and light chains are as follows:
  • the sequence is FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4, the italic in the sequence is the FR sequence; the underline is the CDR sequence, and the double underlined site is the site obtained after affinity maturation screening.
  • the heavy chain amino acid sequence of the PD-L1 antibody or antigen-binding fragment thereof is as shown in SEQ ID NO: 9 or at least 85%, 86% with the sequence shown in SEQ ID NO: 9, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity
  • the PD- The light chain amino acid sequence of the L1 antibody or antigen-binding fragment thereof is as shown in SEQ ID NO: 10 or has at least 85%, 86%, 87%, 88%, 89%, 90% of the sequence shown in SEQ ID NO: 10 , 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity, where
  • PD-L1 antibody heavy chain sequence IgG4(AA)(S228P)
  • Non-limiting example sequences of the TGF- ⁇ RII extracellular domain and truncated forms thereof in the present disclosure are as follows:
  • TGF- ⁇ RII extracellular domain sequence ECD(1-136)
  • Ab is the PD-L1 antibody described in this disclosure.
  • ECD(n-136) is the full-length or truncated form of the extracellular region of TGF- ⁇ RII
  • n is the truncated amino acid of the extracellular region of TGF- ⁇ RII. starting number of digits.
  • PD-L1/TGF- ⁇ fusion protein in WO2018205985A is incorporated herein in its entirety.
  • the heavy chain amino acid sequence of the PD-L1 antibody or its antigen-binding fragment in the fusion protein of TGF- ⁇ receptor is shown in SEQ ID NO:9, and the light chain amino acid sequence is shown in SEQ ID NO:10.
  • the linking sequence L is (G 4 S) 4 G, and the extracellular region of TGF- ⁇ RII is shown in SEQ ID NO: 12.
  • the PD-L1 antibody or antigen-binding fragment thereof is selected from the group consisting of avelumab, atezolizumab, durvalumab, CS-1001, M-7824, KL-A167, CX-072, BGB-A333, GNS-1480, CA-170, BMS-936559, preferably avelumab, atezolizumab, durvalumab.
  • the administered dose of the multi-targeted tyrosine kinase inhibitor is selected from 1-1600 mg, for example: 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg , 65mg, 70mg, 75mg, 80mg, 85mg, 90mg, 95mg, 100mg, 105mg, 110mg, 115mg, 120mg, 125mg, 130mg, 135mg, 140mg, 145mg, 150mg, 155mg, 160mg, 165mg, 170mg, 175mg, 180mg, 185mg , 190mg, 195mg, 200mg, 210mg, 220mg, 230mg, 240mg, 250mg, 260mg, 270mg, 280mg, 290mg, 300mg, 310mg, 320mg,
  • the dosage of the multi-targeted tyrosine kinase inhibitor is selected from 1-800 mg, and the dosing frequency is twice a day, once a day, once every two days, once every three days, Once every four days, once every five days, once every six days, once a week, once every two weeks, once every three weeks or once every four weeks, preferably twice a day or once a day.
  • the multi-targeted tyrosine kinase inhibitor is administered at a dose selected from 5 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 15 mg, 16 mg, 17 mg, 18 mg, 19 mg, 20 mg, 21 mg, 22 mg , 23mg, 24mg, 25mg, 26mg, 27mg, 28mg, 29mg, 30mg, 35mg, 40mg, 45mg, 50mg, 100mg, 150mg, 200mg, 250mg, 300mg, 350mg, 400mg, 450mg, 500mg, 550mg, 600mg, 650mg, 700mg , 750mg, 800mg, the dosing frequency is twice a day, once a day, once every two days, once every three days, once every four days, once every five days, once every six days, once a week, once every two weeks Once, every three weeks or every four weeks, preferably twice a day or once a day or once
  • the administration dose of the fusion protein containing TGF- ⁇ receptor is selected from 0.1-500 mg/kg, and the administration frequency is once a week, once every two weeks, once every three weeks or once every four weeks .
  • the administration dose of the fusion protein containing TGF- ⁇ receptor can be selected from 1 mg/kg, 2 mg/kg, 3 mg/kg, 4 mg/kg, 5 mg/kg, 6 mg/kg, 7 mg/kg, 8 mg/kg , 9mg/kg, 10mg/kg, 11mg/kg, 12mg/kg, 13mg/kg, 14mg/kg, 15mg/kg, 16mg/kg, 17mg/kg, 18mg/kg, 19mg/kg, 20mg/kg, 21mg /kg, 22mg/kg, 23mg/kg, 24mg/kg, 25mg/kg, 26mg/kg, 27mg/kg, 28mg/kg, 29mg/kg, 30mg/kg, 31m
  • the multi-targeted tyrosine kinase inhibitor is selected from c-Kit, VEGFR2, PDGFR inhibitor.
  • the multi-targeted tyrosine kinase inhibitor is a VEGFR2 inhibitor, such as sunitinib, a compound of formula (I) or a pharmaceutically acceptable salt thereof, preferably a compound of formula (I) or a pharmaceutically acceptable salt thereof Salt,
  • the administration dose of the fusion protein containing TGF- ⁇ receptor is selected from 0.1-200 mg/kg, and the administration frequency is once a week, once every two weeks, every three Once a week or every four weeks.
  • the administration dose of the fusion protein containing TGF- ⁇ receptor is selected from 0.1-100 mg/kg, and the administration frequency is once a week, once every two weeks, every three Once a week or every four weeks.
  • the TGF-beta receptor-containing fusion protein (eg, fusion protein 9) is administered at a dose selected from 1 mg/kg, 2 mg/kg, 3 mg/kg, 4 mg/kg, 5 mg/kg, 6 mg /kg, 7mg/kg, 8mg/kg, 9mg/kg, 10mg/kg, 11mg/kg, 12mg/kg, 13mg/kg, 14mg/kg, 15mg/kg, 16mg/kg, 17mg/kg, 18mg/kg , 19mg/kg, 20mg/kg, 21mg/kg, 22mg/kg, 23mg/kg, 24mg/kg, 25mg/kg, 26mg/kg, 27mg/kg, 28mg/kg, 29mg/kg, 30mg/kg, 31mg /kg, 32mg/kg, 33mg/kg, 34mg/kg, 35mg/kg, 36mg/kg, 37mg/kg, 38
  • the TGF-beta receptor-containing fusion protein (eg, fusion protein 9) is administered at a dose selected from 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, 20 mg/kg, 25 mg /kg, 30mg/kg, 35mg/kg, 40mg/kg, 45mg/kg, 50mg/kg, 55mg/kg, 60mg/kg, dosing frequency is once every two weeks, once every three weeks or once every four weeks.
  • the TGF-beta receptor-containing fusion protein (eg, fusion protein 9) is administered at a dose selected from 10 mg/kg, 20 mg/kg, 30 mg/kg, 40 mg/kg, 50 mg/kg, 60 mg /kg, dosing frequency is once every two weeks, once every three weeks or once every four weeks.
  • the TGF-beta receptor-containing fusion protein (eg, fusion protein 9) is administered at a dose of 30 mg/kg at a frequency of once every three weeks.
  • a multi-targeted tyrosine kinase inhibitor (eg, formula (I)) is 20 mg once a day, administered continuously, every 3 weeks (21 days) as a cycle.
  • a fusion protein containing a TGF-beta receptor eg, fusion protein 9
  • a dosing cycle administered on the first day of each cycle.
  • the dose of the fusion protein containing TGF- ⁇ receptor is 1-4000mg, specifically 50mg, 100mg, 150mg, 200mg, 250mg, 300mg, 350mg, 400mg, 450mg , 500mg, 550mg, 600mg, 650mg, 700mg, 750mg, 800mg, 850mg, 900mg, 950mg, 1000mg, 1050mg, 1100mg, 1150mg, 1200mg, 1250mg, 1300mg, 1350mg, 1400mg, 1450mg, 1500mg, 1550mg, 160mg , 1750mg, 1800mg, 1850mg, 1900mg, 1950mg, 2000mg, 2050mg, 2100mg, 2150mg, 2200mg, 2250mg, 2300mg, 2350mg, 2400mg, 2450mg, 2500
  • the dose of the TGF-beta receptor-containing fusion protein is 300 mg, 600 mg, 900 mg, 1200 mg, 1500 mg, 1800 mg, 2100 mg, 2400 mg, 2700 mg, 3000 mg, 3300 mg, 3600 mg, Dosing frequency is every two weeks, every three weeks, or every four weeks.
  • the dose of the TGF-beta receptor-containing fusion protein is 600 mg, 1200 mg, 1800 mg, 2400 mg, 3000 mg, and the dosing frequency is once every two weeks or once every three weeks.
  • the TGF- ⁇ receptor-containing fusion protein (eg, fusion protein 9) is administered by injection, such as subcutaneous or intravenous injection, and the TGF- ⁇ receptor-containing fusion protein needs to be injected before injection.
  • the protein is formulated into an injectable form, and the injectable form of the fusion protein containing the TGF- ⁇ receptor can be an injection or a freeze-dried powder, which comprises the fusion protein containing the TGF- ⁇ receptor, a buffer, and a stabilizer, Optionally also contain surfactants.
  • the buffer can be selected from one or more of acetate, citrate, succinate, and phosphate.
  • Stabilizers may be selected from sugars or amino acids, preferably disaccharides such as sucrose, lactose, trehalose, maltose.
  • the surfactant is selected from polyoxyethylene hydrogenated castor oil, glycerol fatty acid ester, polyoxyethylene sorbitan fatty acid ester and the like.
  • the present disclosure provides a method of treating a tumor or cancer, comprising administering to a subject in need thereof an effective amount of the above-described multi-targeted tyrosine kinase inhibitor and an effective amount of the above-described TGF-beta receptor-containing fusion protein (eg, a fusion protein 9).
  • a method of treating a tumor or cancer comprising administering to a subject in need an effective amount of the above-mentioned multi-targeted tyrosine kinase inhibitor (eg, famitinib or a compound of formula (I) or its pharmaceutically acceptable salt), the multi-targeted tyrosine kinase inhibitor is used in combination with the above-mentioned fusion protein (eg, fusion protein 9) containing the TGF- ⁇ receptor.
  • the above-mentioned multi-targeted tyrosine kinase inhibitor eg, famitinib or a compound of formula (I) or its pharmaceutically acceptable salt
  • the multi-targeted tyrosine kinase inhibitor is used in combination with the above-mentioned fusion protein (eg, fusion protein 9) containing the TGF- ⁇ receptor.
  • a method of treating a tumor or cancer comprising administering to a subject in need thereof an effective amount of the above-described TGF-beta receptor-containing fusion protein (eg, fusion protein 9), the TGF-beta receptor-containing fusion protein
  • the fusion protein is used in combination with the above-mentioned multi-target tyrosine kinase inhibitor (eg, famitinib or the compound represented by formula (I) or a pharmaceutically acceptable salt thereof).
  • the subject in need is a human.
  • the tumors or cancers described in the present disclosure are selected from tumors or cancers of the following sites: colorectal, breast, ovary, pancreas, stomach, prostate, kidney, cervix, myeloma, lymphoma, leukemia, thyroid, endometrium, uterus, Bladder, neuroendocrine, head and neck, liver, nasopharynx, testis, small cell lung cancer, non-small cell lung cancer, melanoma, basal cell skin cancer, squamous cell skin cancer, dermatofibrosarcoma protuberans, meke cell carcinoma, glioblastoma, glioma, sarcoma, mesothelioma, and myelodysplastic syndrome.
  • the tumor or cancer is pancreatic cancer, biliary tract tumor.
  • the biliary tract tumor is a biliary tract malignancy selected from the group consisting of intrahepatic cholangiocarcinoma, extrahepatic cholangiocarcinoma, and gallbladder cancer.
  • the pancreatic cancer is advanced or metastatic pancreatic cancer and the biliary tract tumor (eg, intrahepatic cholangiocarcinoma, extrahepatic cholangiocarcinoma, and gallbladder cancer) is an advanced or metastatic biliary tract tumor.
  • the biliary tract tumor eg, intrahepatic cholangiocarcinoma, extrahepatic cholangiocarcinoma, and gallbladder cancer
  • the biliary tract tumor is selected from biliary tract malignant tumors with mutations in RAS (including KRAS, NRAS, HRAS), BRAF, and PI3KCA genes.
  • RAS including KRAS, NRAS, HRAS
  • BRAF PI3KCA genes.
  • the pancreatic cancer or biliary tract tumor is progressive or metastatic or treatment failure following surgery or first-line therapy.
  • the biliary tract tumor is one or more of uracil, doxorubicin, mitomycin, siggio or capecitabine combination therapy continues to progress or metastasize or Treatment failed.
  • combining refers to the administration of at least one dose of a multitargeted tyrosine kinase inhibitor and at least one dose of a TGF-beta receptor-containing fusion over a period of time proteins, both of which exhibit pharmacological effects.
  • Said time period can be within one dosing cycle, optionally within 4 weeks, within 3 weeks, within 2 weeks, within 1 week, within 24 hours, within 2 hours.
  • the multi-targeted tyrosine kinase inhibitor and the fusion protein containing the TGF-beta receptor can be administered simultaneously or sequentially.
  • This term includes treatment in which the multi-targeted tyrosine kinase inhibitor and the fusion protein containing the TGF-beta receptor are administered by the same route of administration or by different routes of administration.
  • the modes of administration of the combinations described in this disclosure are selected from simultaneous administration, separate formulation and co-administration, or separate formulation and sequential administration.
  • the "antibody” referred to in the present disclosure refers to an immunoglobulin, which is a tetrapeptide chain structure composed of two identical heavy chains and two identical light chains connected by interchain disulfide bonds.
  • the antibody light chain of the present disclosure may further comprise a light chain constant region comprising human or murine ⁇ , ⁇ chains or variants thereof.
  • the antibody heavy chain of the present disclosure may further comprise a heavy chain constant region comprising human or murine IgG1, IgG2, IgG3, IgG4 or variants thereof.
  • variable region The sequence of about 110 amino acids near the N-terminus of the antibody heavy and light chains varies greatly, and is the variable region (Fv region); the remaining amino acid sequences near the C-terminus are relatively stable and are the constant region.
  • the variable region includes three hypervariable regions (HVR) and four relatively conserved framework regions (FR). Three hypervariable regions determine the specificity of antibodies, also known as complementarity determining regions (CDRs).
  • CDRs complementarity determining regions
  • Each light chain variable region (LCVR) and heavy chain variable region (HCVR) consists of 3 CDR regions and 4 FR regions.
  • the sequence from the amino terminus to the carboxy terminus is: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.
  • the three CDR regions of the light chain are referred to as LCDR1, LCDR2, and LCDR3; the three CDR regions of the heavy chain are referred to as HCDR1, HCDR2, and HCDR3.
  • the number and position of CDR amino acid residues in the LCVR and HCVR regions of the antibodies or antigen-binding fragments of the present disclosure conform to the known Kabat numbering conventions (LCDR1-3, HCDE2-3), or the numbering conventions of Kabat and Chothia (HCDR1).
  • Antibodies of the present disclosure include murine antibodies, chimeric antibodies, humanized antibodies, fully human antibodies, eg, humanized antibodies.
  • Antigen-binding fragments include: single-chain antibodies (ie, full-length heavy and light chains); Fab, modified Fab, Fab', modified Fab', F(ab')2, Fv, Fab-Fv, Fab-dsFv, single domain antibodies (eg VH or VL or VHH), scFv, bivalent or trivalent or tetravalent antibodies, Bis-scFv, diabody, tribody, triabody, tetrabody and epitope binding fragments of any of the above (See, eg, Holliger and Hudson, 2005, Nature Biotech. 23(9): 1126-1136; Adair and Lawson, 2005, Drug Design Reviews-Online 2(3), 209-217).
  • Fab-Fv form was first disclosed in WO2009/040562 and its disulfide stabilized form, Fab-dsFv, was first disclosed in WO2010/035012.
  • Antigen-binding fragments of the present disclosure also include Fab and Fab' fragments described in WO2005/003169, WO2005/003170 and WO2005/003171.
  • Multivalent antibodies may comprise multispecific, eg bispecific, or may be monospecific (see eg WO92/22583 and WO05/113605), an example of the latter being the Tri-Fab (or Tri-Fab) described in WO 92/22583 TFM).
  • binding to PD-L1 means capable of interacting with PD-L1 or an epitope thereof, which may be of human origin.
  • antigen-binding site refers to a discrete three-dimensional spatial site on an antigen that is recognized by an antibody or antigen-binding fragment of the present disclosure.
  • murine antibody in the present disclosure is a monoclonal antibody to human PD-L1 prepared according to the knowledge and skill in the art. In preparation, test subjects are injected with PD-L1 antigen, and hybridomas expressing antibodies with desired sequence or functional properties are isolated.
  • chimeric antibody is an antibody obtained by fusing the variable region of a murine antibody with the constant region of a human antibody, which can alleviate the immune response induced by the murine antibody.
  • To establish a chimeric antibody first establish a hybridoma that secretes a mouse-specific monoclonal antibody, then clone the variable region gene from the mouse hybridoma cell, and then clone the constant region gene of the human antibody as needed, and then clone the variable region of the mouse.
  • the gene is linked with the human constant region gene to form a chimeric gene and then inserted into a human vector, and finally the chimeric antibody molecule is expressed in a eukaryotic industrial system or a prokaryotic industrial system.
  • the antibody light chain of the PCSK-9 chimeric antibody further comprises a light chain constant region of a human ⁇ , ⁇ chain or a variant thereof.
  • the antibody heavy chain of the PCSK-9 chimeric antibody further comprises the heavy chain constant region of human IgG1, IgG2, IgG3, IgG4 or a variant thereof.
  • the constant region of the human antibody can be selected from the heavy chain constant region of human IgG1, IgG2, IgG3 or IgG4 or its variants, preferably comprising the heavy chain constant region of human IgG2 or IgG4, or without ADCC (antibody-dependent) after amino acid mutation. cell-mediated cytotoxicity, antibody-dependent cell-mediated cytotoxicity) toxicity of IgG4.
  • humanized antibody also known as CDR-grafted antibody, refers to the grafting of mouse CDR sequences into a human antibody variable region framework, i.e. a different type of human germline Antibody produced in antibody framework sequences.
  • the strong antibody variable antibody response induced by chimeric antibodies can be overcome because they carry a large number of mouse protein components.
  • framework sequences can be obtained from public DNA databases or published references that include germline antibody gene sequences.
  • the germline DNA sequences of human heavy and light chain variable region genes can be found in the "VBase" human germline sequence database, and in Kabat, E.A. et al., 1991 Sequences of Proteins of Immunological Interest, 5th ed.
  • human antibody variable region framework sequence can be subjected to minimal reverse mutation or back mutation to maintain the activity.
  • the humanized antibodies of the present disclosure also include humanized antibodies that are further subjected to affinity maturation of the CDRs by phage display.
  • Identity refers to the sequence similarity between two polynucleotide sequences or between two polypeptides. Sequence identity in the present disclosure may be at least 85%, 90% or 95%, preferably at least 95%. Non-limiting examples include 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% , 100%. Sequence comparison and determination of percent identity between two sequences can be performed with the default settings of the BLASTN/BLASTP algorithm available on the National Center For Biotechnology Institute website.
  • TGF-beta receptor II or “TGFbetaRII” or “transforming growth factor beta receptor II” refers to binding ligands (including but not limited to TGFbeta1, TGFbeta2 and TGFbeta3) and thereby triggering intracellular signal transduction pathway's cell surface receptors.
  • PD-L1 refers to programmed death ligand 1, also known as CD274 and B7H1.
  • PD-L1 is a 290 amino acid protein with extracellular IgV-like and IgC-like domains (amino acids 19-239 of full-length PD-L1), a transmembrane domain, and an intracellular domain of approximately 30 amino acids.
  • PD-L1 is constitutively expressed on many cells such as antigen-presenting cells (eg, dendritic cells, macrophages, and B cells), as well as on hematopoietic and non-hematopoietic cells (eg, vascular endothelial cells, pancreatic islets, and sites of immune forgiveness) .
  • antigen-presenting cells eg, dendritic cells, macrophages, and B cells
  • hematopoietic and non-hematopoietic cells eg, vascular endothelial cells, pancreatic islets, and sites of immune forgiveness
  • PD-L1 is also expressed on a variety of tumors and virus-infected cells and is a component of an immunosuppressive milieu (Ribas 2012, NEJM 366:2517-2519). PD-L1 binds to one of two T-cell co-inhibitors, PD-1 and B7-1.
  • the fusion protein described in the present disclosure is a protein product of co-expression of two genes obtained by DNA recombination.
  • Methods for producing and purifying antibodies and antigen-binding fragments are well known in the art, eg, Cold Spring Harbor's Technical Guide to Antibody Assays, Chapters 5-8 and 15.
  • mice can be immunized with human PD-L1 or fragments thereof, and the resulting antibodies can be renatured, purified, and amino acid sequenced using conventional methods.
  • Antigen-binding fragments can likewise be prepared by conventional methods.
  • the antibody or antigen-binding fragment of the present invention uses genetic engineering to add one or more human FR regions to the non-human CDR regions.
  • Human FR germline sequences can be obtained by aligning the IMGT human antibody variable region germline gene database with MOE software, from the website of ImMunoGeneTics (IMGT) at http://imgt.cines.fr, or from the Journal of Immunoglobulins, 2001 ISBN012441351 get.
  • IMGT ImMunoGeneTics
  • an "effective amount” includes an amount sufficient to ameliorate or prevent a symptom or disorder of a medical disease.
  • An effective amount also means an amount sufficient to allow or facilitate diagnosis.
  • the effective amount for a particular subject in need or veterinary medicine may vary depending on factors such as the condition to be treated, the general health of the subject in need, the method, route and dosage of administration and the severity of side effects.
  • An effective amount can be the maximum dose or dosing regimen that avoids significant side effects or toxic effects.
  • ORR is the objective response rate.
  • PFS progression-free survival.
  • OS is overall survival.
  • DCR is the disease control rate.
  • CR Complete remission
  • Partial remission At least 30% reduction in the sum of target lesion diameters from baseline.
  • PD Disease progression: At least a 20% relative increase in diameter and relative increase in diameter and relative increase of at least 20% (or baseline value if the baseline measurement value is the smallest), with reference to the minimum value of the sum of all measured target lesion diameters throughout the experimental study; otherwise
  • the absolute value of the sum of the diameters must increase by at least 5 mm (the appearance of one or more new lesions is also considered as disease progression).
  • Stable disease The degree of reduction of target lesions does not reach the PR level, and the degree of increase does not reach the level of PD. Between the two, the minimum sum of diameters can be used as a reference in the study.
  • TGF- ⁇ receptor fusion protein fusion protein 9
  • fusion protein 9 multi-target tyrosine kinase inhibitors
  • TGF- ⁇ receptor fusion protein use fusion protein 9, 50mg/mL;
  • Multi-target tyrosine kinase inhibitor compound represented by formula (I).
  • Age 18-75 years old, gender is not limited;
  • TMM stage: IV Histologically or cytologically diagnosed pancreatic cancer or biliary tract tumor (TNM stage: IV); ⁇ 1 line of treatment failure; according to the solid tumor response evaluation criteria (RECIST1.1), imaging diagnosis has at least one measurable lesion.
  • RECIST1.1 Solid tumor response evaluation criteria
  • TGF- ⁇ receptor fusion protein (fusion protein 9): intravenously administered at a dose of 30 mg/kg, administered once every 3 weeks, every 3 weeks (21 days) as a cycle, and administered on the first day of each cycle. It is recommended to administer the drug through an infusion pump, and the intravenous infusion should be completed within 30-60 minutes. Intravenous bolus injection and rapid bolus injection are not allowed. After the infusion is completed, flush the infusion tube with a sufficient amount of normal saline or 5% glucose solution.
  • Multi-targeted tyrosine kinase inhibitor represented by formula (I): 20 mg (initial dose), taken orally before or after meals, recommended daily at a fixed time and within 0.5 hours after meals, once a day; continuous Take the medicine every 3 weeks (21 days) as a cycle.
  • Study drug was used until protocol-specified treatment termination criteria were met or subjects were withdrawn from the study.
  • the primary endpoint was objective response rate (ORR) based on RECIST 1.1 criteria, and secondary endpoints included disease control rate (DCR), progression-free survival (PFS), overall survival (OS), and safety.
  • the safety indicators include vital signs, laboratory indicators, adverse events (AEs), serious adverse events (SAEs), drug-related AEs and SAEs.
  • the incidence of treatment-related adverse events (TRAEs) was 66.67%.
  • fusion protein 9 combined with famitinib had good efficacy and safety in subjects with pancreatic cancer and biliary tract cancer who had failed previous standard treatments. Since the enrolled cases are stage IV subjects in need who have failed at least first-line treatment, CR and PR cases have appeared in a small number of enrolled cases, indicating that the combination administration has a good prospect, and this study is expected to The combination used can extend PFS, OS.

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Abstract

一种TGF-β受体的融合蛋白与多靶点酪氨酸激酶抑制剂联合在制备抗肿瘤药物中的用途。

Description

TGF-β受体的融合蛋白与多靶点酪氨酸激酶抑制剂联合在制备抗肿瘤药物中的用途
本申请要求2020年08月24日提交的中国专利申请(申请号202010854875.3)、2021年04月29日提交的中国专利申请(申请号202110476314.9)和2021年06月16日提交的中国专利申请(申请号202110664907.8)的优先权。本申请引用上述中国专利申请的全文。
技术领域
本公开属于制药领域，具体的涉及一种含有TGF-β受体的融合蛋白与多靶点酪氨酸激酶抑制剂联合在制备治疗肿瘤的药物中的用途。
背景技术
程序性死亡受体1(programmed death 1,PD-1)为CD28超家族成员。PD-1表达于活化的T细胞，B细胞及髓系细胞，其有两个配体，即程序性死亡配体-1(programmed death ligand 1,PD-L1)和PD-L2。PD-L1与T细胞上的受体PD-1相互作用，在免疫应答的负调控方面发挥着重要作用。在许多人类肿瘤组织中均可检测到PD-L1蛋白的表达，肿瘤部位的微环境可诱导肿瘤细胞上的PD-L1的表达，表达的PD-L1有利于肿瘤的发生和生长，诱导抗肿瘤T细胞的凋亡。PD-1/PD-L1通路抑制剂可以阻断PD-1与PD-L1的结合，阻断负向调控信号，使T细胞恢复活性，从而增强免疫应答，因此，以PD-1/PD-L1为靶点的免疫调节对肿瘤抑制有重要的意义。
转化生长因子-β(transforming growth factor-β,TGF-β)属于调节细胞生长和分化的TGF-β超家族。TGF-β通过异源四聚体受体复合物传递信号，这个受体复合物是由两个I型和两个II型的跨膜丝氨酸/苏氨酸激酶受体组成。
研究发现阻断TGF-β信号传导通路能够减少肿瘤的转移。运用截短的Smad2/3显负性突变体抑制乳腺肿瘤细胞系的TGF-β信号通路，结果发现肿瘤细胞的转移能力被抑制。结肠癌的微卫星不稳定性研究发现，TGF-βRII无活性的突变，使转移减少，增加了有需要的受试者术后的存活率。但总体而言，抑制TGF-β信号通路的抑制剂单用在临床治疗中效果微弱，可能跟TGF-β主要在肿瘤细胞内异常性高表达，而单纯的TGF-β信号通路的抑制剂很难集中靶向于肿瘤导致 药效不高或信号通路抑制剂生物利用度有关。
因此，在靶向中和肿瘤微环境的TGF-β基础上抑制PD-1/PD-L1通路，可以使T细胞恢复活性，增强免疫应答，更有效地提高抑治肿瘤发生和发展的效果。Merck公司开发的PD-L1/TGF-β双抗M-7824目前已经进入临床III期，除单药用于治疗肿瘤外，该产品还开发了与艾日布林甲磺酸盐联用治疗转移性三阴性乳腺癌(NCT03579472)；与吉西他滨联用治疗在先接受过治疗的晚期胰腺腺癌(NCT03451773)；与拓扑替康或者替莫唑胺联用治疗复发性小细胞肺癌(NCT0-3554473)等适应症；以及，M7824治疗NSCLC(NCT03631706)已提前终止，M7824联合吉西他滨和顺铂治疗胆道癌(NCT04066491)的II/III期临床试验目前正在招募中。
WO2018205985A公开了一种新的PD-L1/TGF-β融合蛋白。CN101007815A提供了多靶点酪氨酸激酶抑制剂，包括如式(I)化合物，并公开了该化合物具有VEGF、EGFR抑制活性。CN106535896A公开了式(I)化合物可用于治疗纤维化疾病，例如肺纤维化、肝硬化、硬皮病或者肾纤维化。本公开全文引入上述专利内容。
Figure PCTCN2021114280-appb-000001
胰腺癌和胆道恶性肿瘤恶性程度较高、进展迅速,但起病隐匿、早期症状不典型，临床就诊时大部分有需要的受试者已属于中晚期。胰腺癌和胆道恶性肿瘤的治疗是一个系统性的工程，常常需要多学科会诊模式(MDT)，主要包括手术治疗、放射治疗、化学治疗、介入治疗和支持治疗，其中手术切除是胰腺癌和胆道恶性肿瘤有需要的受试者获得治愈和长期生存的唯一机会。对于一线治疗失败的晚期胰腺癌，可选的治疗方案有限，根据有需要的受试者的身体功能状态及肿瘤进展情况，可以选择以吉西他滨或者氟尿嘧啶为基础的单药化疗或者联合化疗，二线化疗ORR<10％。对于一线治疗失败的晚期胆道恶性肿瘤，首选的化疗方案为奥沙利铂联合5-FU，免疫治疗方案包括帕博丽珠单抗(Pembrolizumab)联合仑伐替尼、帕博丽珠单抗(仅高微卫星不稳定(MSI-H)/错配修复缺陷(dMMR) 类型的肿瘤)及纳武利尤单抗已获得中国临床肿瘤学会(CSCO)及美国国立综合癌症网络(NCCN)指南推荐，但目前尚无Ⅲ期研究数据公布，免疫治疗疗效还需进一步探索。
因此，寻找更为有效的治疗新途径或药物组合，为胰腺癌、胆道肿瘤临床治疗提供新的循证医学证据，从而指导有需要的受试者的临床诊疗综合当前国际国内研究新进展，具有重要的临床意义。
发明内容
本公开涉及一种含有TGF-β受体的融合蛋白与多靶点酪氨酸激酶抑制剂联合在制备治疗肿瘤或癌症的药物中的用途。
本公开提供一种治疗、缓解或预防肿瘤或癌症的方法，包括向有需要的受试者施用治疗、缓解或预防有效量的多靶点酪氨酸激酶抑制剂和治疗、缓解或预防有效量的含有TGF-β受体的融合蛋白。
一些实施方案中，提供治疗肿瘤或癌症的方法，包括有需要的受试者施用治疗、缓解或预防有效量的上述多靶点酪氨酸激酶抑制剂，所述多靶点酪氨酸激酶抑制剂与所述含有TGF-β受体的融合蛋白联用。
一些实施方案中，提供治疗肿瘤或癌症的方法，包括有需要的受试者施用治疗、缓解或预防有效量的含有TGF-β受体的融合蛋白，所述含有TGF-β受体的融合蛋白与所述多靶点酪氨酸激酶抑制剂联用。
在某些实施方式中，所述的多靶点酪氨酸激酶抑制剂为式(I)化合物或其可药用盐，
Figure PCTCN2021114280-appb-000002
在某些实施方式中，所述的多靶点酪氨酸激酶抑制剂为法米替尼或其可药用盐。
在某些实施方式中，所述的TGF-β受体的融合蛋白包含PD-L1抗体或其抗原结合片段和TGF-β受体，其中TGF-β受体部分为TGF-βRII胞外区的N端截短形式。
在某些实施方式中，所述TGF-β受体融合蛋白如通式(II)所示：
Ab-L-TGF-βRII ECD                 (II)
其中TGF-βRII ECD为TGF-βRII胞外区的截短形式；
Ab为PD-L1抗体或其抗原结合片段；
L为连接序列。
在某些实施方式中，所述的连接序列为(G 4S)xG，其中x为3-6，例如为4或5。
在某些实施方式中，所述的PD-L1抗体或其抗原结合片段包含：
分别如SEQ ID NO:1、SEQ ID NO:2和SEQ ID NO:3所示的HCDR1、HCDR2和HCDR3，和分别如SEQ ID NO:4、SEQ ID NO:5和SEQ ID NO:6所示的LCDR1、LCDR2和LCDR3，
其中，前面所述的各CDR序列如下所示：
HCDR1：SYWMH                      SEQ ID NO:1
HCDR2：RIGPNSGFTSYNEKFKN          SEQ ID NO:2
HCDR3：GGSSYDYFDY                 SEQ ID NO:3
LCDR1：RASESVSIHGTHLMH            SEQ ID NO:4
LCDR2：AASNLES                    SEQ ID NO:5
LCDR3：QQSFEDPLT                  SEQ ID NO:6
在某些实施方式中，所述PD-L1抗体或其抗原结合片段谓嵌合抗体或其功能片段、人源化抗体或其功能片段或人抗体或其功能片段。
在某些实施方式中，所述人源化PD-L1抗体重、轻链的序列如下所示：
PD-L1抗体重链可变区：
Figure PCTCN2021114280-appb-000003
PD-L1抗体轻链可变区：
Figure PCTCN2021114280-appb-000004
注：顺序为FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4，序列中斜体为FR序列；下划线为CDR序列，其中双下划线的位点为亲和力成熟筛选后得到的位点。
在某些实施方案中，所述PD-L1抗体或其抗原结合片段的重链氨基酸序列如SEQ ID NO:9所示或与SEQ ID NO:9所示的序列具有至少85％，86％，87％，88％，89％，90％，91％，92％，93％，94％，95％，96％，97％，98％，99％或100％的序列同一性，所述PD-L1抗体或其抗原结合片段的轻链氨基酸序列如SEQ ID NO:10所示或与SEQ ID NO:10所示的序列具有至少85％，86％，87％，88％，89％，90％，91％，92％，93％，94％，95％，96％，97％，98％，99％或100％的序列同一性，其中
PD-L1抗体重链序列：Ig G4(AA)(S228P)
Figure PCTCN2021114280-appb-000005
PD-L1抗体轻链序列：
Figure PCTCN2021114280-appb-000006
本公开中TGF-βRII胞外结构域及其截短形式的非限制性实施例序列如下：
TGF-βRII胞外结构域序列：ECD(1-136)
Figure PCTCN2021114280-appb-000007
Figure PCTCN2021114280-appb-000008
TGF-βRII胞外结构域序列在N端有19个氨基酸的截短或缺失：ECD(20-136)
Figure PCTCN2021114280-appb-000009
TGF-βRII胞外结构域序列在N端有21个氨基酸的截短或缺失：ECD(22-136)
Figure PCTCN2021114280-appb-000010
TGF-βRII胞外结构域序列在N端有14个氨基酸的截短或缺失：ECD(15-136)
Figure PCTCN2021114280-appb-000011
利用同源重组技术将PD-L1抗体的重链C末端氨基酸通过(G 4S) xG连接不同长度TGF-βRII胞外区，与轻链一起，通过293表达系统进行常规表达，得到如表1所示的融合蛋白：
表1.PD-L1抗体/TGF-βRII胞外区融合蛋白
Figure PCTCN2021114280-appb-000012
Figure PCTCN2021114280-appb-000013
注：Ab为本公开所述PD-L1抗体，序列描述中ECD(n-136)为TGF-βRII胞外区的全长或截短形式，n为TGF-βRII胞外区截短后的氨基酸起始位数。
此处全文引入WO2018205985A中的PD-L1/TGF-β融合蛋白。
一些具体实施方案中，TGF-β受体的融合蛋白中的PD-L1抗体或其抗原结合片段的重链氨基酸序列如SEQ ID NO:9所示、轻链氨基酸序列如SEQ ID NO:10所示，连接序列L为(G 4S) 4G，TGF-βRII胞外区如SEQ ID NO:12所示。
在一些实施方式中，所述PD-L1抗体或其抗原结合片段选自avelumab、atezolizumab、durvalumab、CS-1001、M-7824、KL-A167、CX-072、、BGB-A333、GNS-1480、CA-170、BMS-936559，优选avelumab、atezolizumab、durvalumab。
在一些实施方式中，所述多靶点酪氨酸激酶抑制剂的给药剂量选自1-1600mg，例如：10mg、15mg、20mg、25mg、30mg、35mg、40mg、45mg、50mg、55mg、60mg、65mg、70mg、75mg、80mg、85mg、90mg、95mg、100mg、105mg、110mg、115mg、120mg、125mg、130mg、135mg、140mg、145mg、150mg、155mg、160mg、165mg、170mg、175mg、180mg、185mg、190mg、195mg、200mg、210mg、220mg、230mg、240mg、250mg、260mg、270mg、280mg、290mg、300mg、310mg、320mg、330mg、340mg、350mg、360mg、370mg、380mg、390mg、400mg、410mg、420mg、430mg、440mg、450mg、460mg、470mg、480mg、490mg、500mg、510mg、520mg、530mg、540mg、550mg、560mg、570mg、580mg、590mg、600mg、625mg、650mg、675mg、700mg、725mg、750mg、775mg、800mg、825mg、850mg、875mg、900mg、925mg、950mg、975mg、1000mg、1025mg、1050mg、1075mg、1100mg、1125mg、1150mg、1175mg、1200mg、1225mg、1250mg、1275mg、1300mg、1325mg、1350mg、1375mg、1400mg、1425mg、1450mg、1475mg、1500mg、1525mg、1550mg、1575mg、1600mg，给药频率为一日两次，一日一次，每两日一次，每三日一次，每四日一次，每五日一次，每六日一次，每周一次，每两周一次，每三周一次或 每四周一次，优选一日两次或一日一次。
在一些实施方式中，所述多靶点酪氨酸激酶抑制剂的给药剂量选自1-800mg，给药频率为一日两次，一日一次，每两日一次，每三日一次，每四日一次，每五日一次，每六日一次，每周一次，每两周一次，每三周一次或每四周一次，优选一日两次或一日一次。
在一些实施方式中，所述多靶点酪氨酸激酶抑制剂的给药剂量选自5mg、10mg、11mg、12mg、13mg、14mg、15mg、16mg、17mg、18mg、19mg、20mg、21mg、22mg、23mg、24mg、25mg、26mg、27mg、28mg、29mg、30mg、35mg、40mg、45mg、50mg、100mg、150mg、200mg、250mg、300mg、350mg、400mg、450mg、500mg、550mg、600mg、650mg、700mg、750mg、800mg，给药频率为一日两次，一日一次，每两日一次，每三日一次，每四日一次，每五日一次，每六日一次，每周一次，每两周一次，每三周一次或每四周一次，优选一日两次或一日一次。
在一些实施方式中，所述含有TGF-β受体的融合蛋白的给药剂量选自0.1-500mg/kg，给药频率为每周一次，每两周一次，每三周一次或每四周一次。具体的,含有TGF-β受体的融合蛋白的给药剂量可以选自1mg/kg、2mg/kg、3mg/kg、4mg/kg、5mg/kg、6mg/kg、7mg/kg、8mg/kg、9mg/kg、10mg/kg、11mg/kg、12mg/kg、13mg/kg、14mg/kg、15mg/kg、16mg/kg、17mg/kg、18mg/kg、19mg/kg、20mg/kg、21mg/kg、22mg/kg、23mg/kg、24mg/kg、25mg/kg、26mg/kg、27mg/kg、28mg/kg、29mg/kg、30mg/kg、31mg/kg、32mg/kg、33mg/kg、34mg/kg、35mg/kg、36mg/kg、37mg/kg、38mg/kg、39mg/kg、40mg/kg、41mg/kg、42mg/kg、43mg/kg、44mg/kg、45mg/kg、46mg/kg、47mg/kg、48mg/kg、49mg/kg、50mg/kg、51mg/kg、52mg/kg、53mg/kg、54mg/kg、55mg/kg、56mg/kg、57mg/kg、58mg/kg、59mg/kg、60mg/kg、61mg/kg、62mg/kg、63mg/kg、64mg/kg、65mg/kg、66mg/kg、67mg/kg、68mg/kg、69mg/kg、70mg/kg、71mg/kg、72mg/kg、73mg/kg、74mg/kg、75mg/kg、76mg/kg、77mg/kg、78mg/kg、79mg/kg、80mg/kg、81mg/kg、82mg/kg、83mg/kg、84mg/kg、85mg/kg、86mg/kg、87mg/kg、88mg/kg、89mg/kg、90mg/kg、91mg/kg、92mg/kg、93mg/kg、94mg/kg、95mg/kg、96mg/kg、97mg/kg、98mg/kg、99mg/kg、100mg/kg、105mg/kg、110mg/kg、115mg/kg、120mg/kg、125mg/kg、130mg/kg、135mg/kg、140mg/kg、145mg/kg、150mg/kg、155mg/kg、160mg/kg、165mg/kg、170mg/kg、175mg/kg、180mg/kg、185mg/kg、190mg/kg、195mg/kg、200mg/kg、205mg/kg、 210mg/kg、215mg/kg、220mg/kg、225mg/kg、230mg/kg、235mg/kg、240mg/kg、245mg/kg、250mg/kg、260mg/kg、270mg/kg、280mg/kg、290mg/kg、300mg/kg、310mg/kg、320mg/kg、330mg/kg、340mg/kg、350mg/kg、360mg/kg、370mg/kg、380mg/kg、390mg/kg、400mg/kg、410mg/kg、420mg/kg、430mg/kg、440mg/kg、450mg/kg、460mg/kg、470mg/kg、480mg/kg、490mg/kg、500mg/kg。
上述实施方式中，多靶点酪氨酸激酶抑制剂选自c-Kit、VEGFR2、PDGFR抑制剂。一些具体实施方案中，多靶点酪氨酸激酶抑制剂为VEGFR2抑制剂，例如舒尼替尼、式(I)化合物或其可药用盐，优选为式(I)化合物或其可药用盐，
在一些实施方式中，所述含有TGF-β受体的融合蛋白(例如融合蛋白9)的给药剂量选自0.1-200mg/kg，给药频率为每周一次，每两周一次，每三周一次或每四周一次。
在一些实施方式中，所述含有TGF-β受体的融合蛋白(例如融合蛋白9)的给药剂量选自0.1-100mg/kg，给药频率为每周一次，每两周一次，每三周一次或每四周一次。
在一些实施方式中，所述含有TGF-β受体的融合蛋白(例如融合蛋白9)的给药剂量选自1mg/kg、2mg/kg、3mg/kg、4mg/kg、5mg/kg、6mg/kg、7mg/kg、8mg/kg、9mg/kg、10mg/kg、11mg/kg、12mg/kg、13mg/kg、14mg/kg、15mg/kg、16mg/kg、17mg/kg、18mg/kg、19mg/kg、20mg/kg、21mg/kg、22mg/kg、23mg/kg、24mg/kg、25mg/kg、26mg/kg、27mg/kg、28mg/kg、29mg/kg、30mg/kg、31mg/kg、32mg/kg、33mg/kg、34mg/kg、35mg/kg、36mg/kg、37mg/kg、38mg/kg、39mg/kg、40mg/kg、41mg/kg、42mg/kg、43mg/kg、44mg/kg、45mg/kg、46mg/kg、47mg/kg、48mg/kg、49mg/kg、50mg/kg、51mg/kg、52mg/kg、53mg/kg、54mg/kg、55mg/kg、56mg/kg、57mg/kg、58mg/kg、59mg/kg、60mg/kg，给药频率为每两周一次，每三周一次或每四周一次。
在一些实施方式中，所述含有TGF-β受体的融合蛋白(例如融合蛋白9)的给药剂量选自1mg/kg、3mg/kg、10mg/kg、15mg/kg、20mg/kg、25mg/kg、30mg/kg、35mg/kg、40mg/kg、45mg/kg、50mg/kg、55mg/kg、60mg/kg，给药频率为每两周一次，每三周一次或每四周一次。
在一些实施方式中，所述含有TGF-β受体的融合蛋白(例如融合蛋白9)的给药剂量选自10mg/kg、20mg/kg、30mg/kg、40mg/kg、50mg/kg、60mg/kg，给 药频率为每两周一次，每三周一次或每四周一次。
在一些实施方式中，所述含有TGF-β受体的融合蛋白(例如融合蛋白9)的给药剂量为30mg/kg，给药频率为每三周一次。多靶点酪氨酸激酶抑制剂(例如式(I))是20mg，每日1次，连续服药，每3周(21天)为一个周期。
一些实施方式中，当每周一次、每两周一次、每三周一次或每四周一次给予有需要的受试者或受试者含有TGF-β受体的融合蛋白(例如融合蛋白9)时，每周一次、每两周一次、每三周一次或每四周一次分别为一个给药周期，在每个周期的第1天给药。
在一些实施方式中，所述含有TGF-β受体的融合蛋白(例如融合蛋白9)的剂量为1-4000mg，具体可以为50mg、100mg、150mg、200mg、250mg、300mg、350mg、400mg、450mg、500mg、550mg、600mg、650mg、700mg、750mg、800mg、850mg、900mg、950mg、1000mg、1050mg、1100mg、1150mg、1200mg、1250mg、1300mg、1350mg、1400mg、1450mg、1500mg、1550mg、1600mg、1650mg、1700mg、1750mg、1800mg、1850mg、1900mg、1950mg、2000mg、2050mg、2100mg、2150mg、2200mg、2250mg、2300mg、2350mg、2400mg、2450mg、2500mg、2550mg、2600mg、2650mg、2700mg、2750mg、2800mg、2850mg、2900mg、2950mg、3000mg、3050mg、3100mg、3150mg、3200mg、3250mg、3300mg、3350mg、3400mg、3450mg、3500mg、3550mg、3600mg、3650mg、3700mg、3750mg、3800mg、3850mg、3900mg、3950mg、4000mg，给药频率为每周一次，每两周一次，每三周一次或每四周一次。
在一些实施方式中，所述含有TGF-β受体的融合蛋白(例如融合蛋白9)的剂量为300mg、600mg、900mg、1200mg、1500mg、1800mg、2100mg、2400mg、2700mg、3000mg、3300mg、3600mg，给药频率为每两周一次，每三周一次或每四周一次。
在一些实施方式中，所述含有TGF-β受体的融合蛋白(例如融合蛋白9)的剂量为600mg、1200mg、1800mg、2400mg、3000mg，给药频率为每两周一次或每三周一次。
可选的实施方案中，所述的含有TGF-β受体的融合蛋白(例如融合蛋白9)以注射的方式给药，例如皮下或静脉注射，注射前需将含有TGF-β受体的融合蛋白配制成可注射的形式，含有TGF-β受体的融合蛋白的可注射形式可以是是注射液或冻干粉针，其包含含有TGF-β受体的融合蛋白、缓冲剂、稳定剂，任选地还含有表面活性剂。缓冲剂可选自醋酸盐、柠檬酸盐、琥珀酸盐、以及磷酸盐 中的一种或几种。稳定剂可选自糖或氨基酸，优选二糖，例如蔗糖、乳糖、海藻糖、麦芽糖。表面活性剂选自聚氧乙烯氢化蓖麻油、甘油脂肪酸酯、聚氧乙烯山梨醇酐脂肪酸酯等。
本公开提供一种治疗肿瘤或癌症的方法，包括向有需要的受试者施用有效量上述多靶点酪氨酸激酶抑制剂和有效量上述含有TGF-β受体的融合蛋白(例如融合蛋白9)。
一些实施方案中，提供治疗肿瘤或癌症的方法，包括有需要的受试者施用有效量的上述多靶点酪氨酸激酶抑制剂(例如法米替尼或式(I)所示化合物或其可药用盐)，所述多靶点酪氨酸激酶抑制剂与上述含有TGF-β受体的融合蛋白(例如融合蛋白9)联用。
一些实施方案中，提供治疗肿瘤或癌症的方法，包括有需要的受试者施用有效量的上述含有TGF-β受体的融合蛋白(例如融合蛋白9)，所述含有TGF-β受体的融合蛋白与上述多靶点酪氨酸激酶抑制剂(例如法米替尼或式(I)所示化合物或其可药用盐)联用。
可选的实施方案中，前述治疗肿瘤或癌症的方法或用途中，所述有需要的受试者为人类。
本公开中所述肿瘤或癌症选自以下部位的肿瘤或癌症：结直肠、乳腺、卵巢、胰腺、胃、前列腺、肾、宫颈、骨髓瘤、淋巴瘤、白血病、甲状腺、子宫内膜、子宫、膀胱、神经内分泌、头部颈部、肝、鼻咽、睾丸、小细胞肺癌、非小细胞肺癌、黑素瘤、基底细胞皮肤癌、鳞状细胞皮肤癌、隆突性皮肤纤维肉瘤、梅克尔细胞癌、成胶质细胞瘤、胶质瘤、肉瘤、间皮瘤，和骨髓增生异常综合征。
一些具体实施方案中，所述肿瘤或癌症为胰腺癌、胆道肿瘤。
一些具体实施方案中，所述胆道肿瘤为胆道恶性肿瘤，选自肝内胆管癌、肝外胆管癌和胆囊癌。
一些具体实施方案中，所述胰腺癌为晚期或转移性胰腺癌，所述胆道肿瘤(例如肝内胆管癌、肝外胆管癌和胆囊癌)是晚期或转移性胆道肿瘤。
一些具体实施方案中，所述胆道肿瘤选自RAS(包括KRAS、NRAS、HRAS)、BRAF、PI3KCA基因突变的胆道恶性肿瘤。
一些具体实施方案中，所述胰腺癌或胆道肿瘤是经过手术治疗或一线治疗的继续进展的或转移的或治疗失败的。
一些具体实施方案中，所述胆道肿瘤是接受过尿嘧啶类、阿霉素、丝裂霉素、 替吉奥或者卡培他滨中一种或多种组合治疗后继续进展的或转移的或治疗失败的。
具体实施方式
术语
本公开中所述“联合”一种给药方式，是指在一定时间期限内给予至少一种剂量的多靶点酪氨酸激酶抑制剂和至少一种剂量的含有TGF-β受体的融合蛋白，其中两种物质都显示药理学作用。所述的时间期限可以是一个给药周期内，可选4周内，3周内，2周内，1周内，24小时以内，2小时以内。可以同时或依次给予多靶点酪氨酸激酶抑制剂和含有TGF-β受体的融合蛋白。这种期限包括这样的治疗，其中通过相同给药途径或不同给药途径给予多靶点酪氨酸激酶抑制剂和含有TGF-β受体的融合蛋白。本公开所述联合的给药方式选自同时给药、独立地配制并共给药或独立地配制并相继给药。
本公开所述的“抗体”指免疫球蛋白，是由两条相同的重链和两条相同的轻链通过链间二硫键连接而成的四肽链结构。
在本公开中，本公开所述的抗体轻链可进一步包含轻链恒定区，所述的轻链恒定区包含人源或鼠源的κ、λ链或其变体。
在本公开中，本公开所述的抗体重链可进一步包含重链恒定区，所述的重链恒定区包含人源或鼠源的IgG1、IgG2、IgG3、IgG4或其变体。
抗体重链和轻链靠近N端的约110个氨基酸的序列变化很大，为可变区(Fv区)；靠近C端的其余氨基酸序列相对稳定，为恒定区。可变区包括3个高变区(HVR)和4个序列相对保守的骨架区(FR)。3个高变区决定抗体的特异性，又称为互补性决定区(CDR)。每条轻链可变区(LCVR)和重链可变区(HCVR)由3个CDR区4个FR区组成，从氨基端到羧基端依次排列的顺序为：FR1，CDR1，FR2，CDR2，FR3，CDR3，FR4。轻链的3个CDR区指LCDR1、LCDR2、和LCDR3；重链的3个CDR区指HCDR1、HCDR2和HCDR3。本公开所述的抗体或抗原结合片段的LCVR区和HCVR区的CDR氨基酸残基在数量和位置符合已知的Kabat编号规则(LCDR1-3，HCDE2-3)，或者符合kabat和chothia的编号规则(HCDR1)。
本公开的抗体包括鼠源抗体、嵌合抗体、人源化抗体、全人抗体，例如人源化抗体。
本公开“抗原结合片段”包括：单链抗体(即全长重链和轻链)；Fab、修饰的Fab、Fab’、修饰的Fab’、F(ab’)2、Fv、Fab-Fv、Fab-dsFv、单结构域抗体(例如VH或VL或VHH)、scFv、二价或三价或四价抗体、Bis-scFv、diabody、tribody、triabody、tetrabody和上述任意一种的表位结合片段(参见例如Holliger and Hudson,2005,Nature Biotech.23(9):1126-1136；Adair and Lawson,2005,Drug Design Reviews-Online 2(3),209-217)。产生和制备这些抗体片段的方法在本领域是公知的(参见例如Verma等人,1998,Journal ofImmunological Methods,216,165-181)。Fab-Fv形式首先公开于WO2009/040562，其二硫键稳定化形式Fab-dsFv首先公开于WO2010/035012。本公开的抗原结合片段还包括描述于WO2005/003169、WO2005/003170和WO2005/003171中的Fab和Fab’片段。多价抗体可包含多特异性例如双特异性或可以是单特异性的(参见例如WO92/22583和WO05/113605)，后者的一个示例是描述于WO 92/22583中的Tri-Fab(或TFM)。
术语“与PD-L1结合”，指能与PD-L1或其表位相互作用，所述PD-L1或其表位可以是人源的。
术语“抗原结合位点”指抗原上不连续的，由本公开抗体或抗原结合片段识别的三维空间位点。
术语“鼠源抗体”在本公开中为根据本领域知识和技能制备的对人PD-L1的单克隆抗体。制备时用PD-L1抗原注射试验对象，然后分离表达具有所需序列或功能特性的抗体的杂交瘤。
术语“嵌合抗体(chimeric antibody)”，是将鼠源性抗体的可变区与人抗体的恒定区融合而成的抗体，可以减轻鼠源性抗体诱发的免疫应答反应。建立嵌合抗体，要先建立分泌鼠源性特异性单抗的杂交瘤，然后从小鼠杂交瘤细胞中克隆可变区基因，再根据需要克隆人抗体的恒定区基因，将小鼠可变区基因与人恒定区基因连接成嵌合基因后插入人载体中，最后在真核工业系统或原核工业系统中表达嵌合抗体分子。在本公开一个优选的实施方案中，所述的PCSK-9嵌合抗体的抗体轻链进一步包含人源κ、λ链或其变体的轻链恒定区。所述的PCSK-9嵌合抗体的抗体重链进一步包含人源IgG1、IgG2、IgG3、IgG4或其变体的重链恒定区。人抗体的恒定区可选自人源IgG1、IgG2、IgG3或IgG4或其变体的重链恒定区，优选包含人源IgG2或IgG4重链恒定区，或者使用氨基酸突变后无ADCC(antibody-dependent cell-mediated cytotoxicity，抗体依赖的细胞介导的细胞毒作用)毒性的IgG4。
术语“人源化抗体(humanized antibody)”，也称为CDR移植抗体(CDR-grafted antibody)，是指将小鼠的CDR序列移植到人的抗体可变区框架，即不同类型的人种系抗体构架序列中产生的抗体。可以克服嵌合抗体由于携带大量小鼠蛋白成分，从而诱导的强烈的抗体可变抗体反应。此类构架序列可以从包括种系抗体基因序列的公共DNA数据库或公开的参考文献获得。如人重链和轻链可变区基因的种系DNA序列可以在“VBase”人种系序列数据库，以及在Kabat，E.A.等人，1991 Sequences of Proteins of Immunological Interest，第5版中找到。为避免免疫原性下降的同时，引起的活性下降，可对所述的人抗体可变区框架序列进行最少反向突变或回复突变，以保持活性。本公开的人源化抗体也包括进一步由噬菌体展示对CDR进行亲和力成熟后的人源化抗体。
本公开中所述“同一性”是指两个多核苷酸序列之间或两个多肽之间的序列相似性。本公开中的序列同一性可以至少为85％、90％或95％，优选至少为95％。非限制性实施例包括85％，86％，87％，88％，89％，90％，91％，92％，93％，94％，95％，96％，97％，98％，99％，100％。两个序列之间的序列比较和同一性百分比测定可以通过National Center For Biotechnology Institute网站上可得的BLASTN/BLASTP算法的默认设置来进行。
术语“TGF-β受体II”或“TGFβRII”或“转化生长因子β受体II”是指结合配体(包括但不限于TGFβ1、TGFβ2和TGFβ3)、并且由此引发细胞内的信号转导途径的细胞表面受体。
术语“PD-L1”是指程序性死亡配体1，也称为CD274和B7H1。PD-L1是具有胞外IgV样和IgC样结构域(全长PD-L1的氨基酸19-239)、跨膜结构域和约30个氨基酸的胞内结构域的290个氨基酸的蛋白质。PD-L1在许多细胞例如抗原呈递细胞(例如，树突细胞、巨噬细胞和B细胞)上以及造血细胞和非造血细胞(例如，血管内皮细胞、胰岛、和免疫赦免部位)上组成型表达。PD-L1也在多种肿瘤和病毒感染的细胞上表达，并且是免疫抑制环境(immunosuppressive milieu)的组成(Ribas 2012，NEJM 366：2517-2519)。PD-L1与两种T细胞共抑制剂PD-1和B7-1之一结合。
本公开中所述的融合蛋白是一种通过DNA重组得到的两个基因共表达的蛋白产物。现有技术中熟知生产和纯化抗体和抗原结合片段的方法，如冷泉港的抗体实验技术指南，5-8章和15章。例如，小鼠可以用人PD-L1或其片段免疫，所得到的抗体能被复性、纯化，并且可以用常规的方法进行氨基酸测序。抗原结 合片段同样可以用常规方法制备。发明所述的抗体或抗原结合片段用基因工程方法在非人源的CDR区加上一个或多个人源FR区。人FR种系序列可以通过比对IMGT人类抗体可变区种系基因数据库和MOE软件，从ImMunoGeneTics(IMGT)的网站http://imgt.cines.fr得到，或者从免疫球蛋白杂志，2001ISBN012441351上获得。
“有效量”包含足以改善或预防医学疾病的症状或病症的量。有效量还意指足以允许或促进诊断的量。用于特定有需要的受试者或兽医学受试者的有效量可依据以下因素而变化：例如，待治疗的病症、有需要的受试者的总体健康情况、给药的方法途径和剂量以及副作用严重性。有效量可以是避免显著副作用或毒性作用的最大剂量或给药方案。
ORR为客观缓解率。PFS为无进展生存期。OS为总生存期。DCR为疾病控制率。
完全缓解(CR)：所有靶病灶消失，全部病理淋巴结(包括靶结节和非靶结节)短直径必须减少至＜10mm。
部分缓解(PR)：靶病灶直径之和比基线水平减少至少30％。
疾病进展(PD)：以整个实验研究过程中所有测量的靶病灶直径之和的最小值为参照，直径和相对增加至少20％(如果基线测量值最小就以基线值为参照)；除此之外，必须满足直径和的绝对值增加至少5mm(出现一个或多个新病灶也视为疾病进展)。
疾病稳定(SD)：靶病灶减小的程度没达到PR，增加的程度也没达到PD水平，介于两者之间，研究时可以直径之和的最小值作为参考。
以下结合实施例用于进一步描述本发明，但这些实施例并非限制本公开的范围。
实施例评价测试药物在人体内的药效
分别招募20-30例晚期胰腺癌及胆道肿瘤受试者，初步评估TGF-β受体融合蛋白(融合蛋白9)联合多靶点酪氨酸激酶抑制剂在晚期胰腺癌及胆道肿瘤中的疗效。
1、受试抗体和化合物
TGF-β受体融合蛋白：使用融合蛋白9，50mg/mL；
多靶点酪氨酸激酶抑制剂：式(I)所示化合物。
2、入组受试者标准
1)年龄18-75岁，性别不限；
2)组织学或细胞学确诊为胰腺癌或胆道肿瘤(TNM分期：IV期)；≥1线治疗失败；根据实体瘤缓解评价标准(RECIST1.1)，影像学诊断至少有一个可测量病灶。
3、给药方法
TGF-β受体融合蛋白(融合蛋白9)：静脉给药，剂量30mg/kg，3周给药1次，每3周(21天)为一周期，每周期第1天给药。建议通过输液泵给药，静脉输注30-60分钟完成给药，不得静脉推注和快速推注，输液结束后，用足量的生理盐水或者5％的葡萄糖溶液冲洗输液管。
式(I)所示多靶点酪氨酸激酶抑制剂：20mg(起始剂量)，餐前或餐后口服，推荐每日固定时间、餐后0.5h内给药，每日1次；连续服药，每3周(21天)为一个周期。
使用研究药物直至出现方案规定的治疗终止标准或受试者退出研究。
受试者出现符合实体瘤的疗效评价标准第1.1版(RECIST1.1)定义的进展后，如果研究者评估受试者临床获益并可耐受研究治疗且有需要的受试者自愿的前提下，仍可继续TGF-β受体融合蛋白和/或式(I)所示多靶点酪氨酸激酶抑制剂治疗。
主要的评价终点是基于RECIST 1.1标准评估的客观缓解率(ORR)，次要终点包括疾病控制率(DCR)、无进展生存期(PFS)、总生存期(OS)和安全性。所述安全性的指标包括生命体征、实验室指标、不良事件(AE)、严重不良事件(SAE)、药物相关性AE及SAE。
共入组18例胰腺癌和6例胆道癌。胰腺癌队列可评估的12例有需要的受试者中，1例CR，4例SD，7例PD，ORR 8.3％，DCR 41.6％；胆道癌队列已评估3例，包括1例PR(持续3.3+个月，肿瘤缩小46.48％)，1例SD，ORR 33.3％，DCR 66.7％。治疗相关不良事件(TRAE)发生率66.67％，最常见TRAE为高血压(38.10％)、蛋白尿(38.10％)和丙氨酸氨基转移酶升高(33.33％)，最常见3级TRAE为血胆红素升高(9.52％)。无4/5级AE发生。
结果显示，在既往标准治疗失败的胰腺癌和胆道癌有需要的受试者中，融合蛋白9联合法米替尼治疗具有较好的疗效和安全性。由于在入组的病例是已经经过至少一线治疗失败的IV期有需要的受试者，在少量入组病例中已经出现CR 和PR的病例，说明该联合给药具有良好的前景，预期本研究使用的组合可延长PFS、OS。

Claims (19)

  1. 一种含有TGF-β受体的融合蛋白与多靶点酪氨酸激酶抑制剂联合在制备治疗肿瘤或癌症的药物中的用途，所述肿瘤或癌症优选为胰腺癌、胆道肿瘤；所述胆道肿瘤优选自肝内胆管癌、肝外胆管癌和胆囊癌。
  2. 根据权利要求1所述的用途，其特征在于，所述胰腺癌、胆道肿瘤为晚期或转移性的。
  3. 根据权利要求1或2所述的用途，其特征在于，所述的TGF-β受体的融合蛋白包含PD-L1抗体或其抗原结合片段和TGF-β受体，其中TGF-β受体部分为TGF-βRII胞外区的N端截短形式。
  4. 根据权利要求3所述的用途，其特征在于，所述的TGF-β受体融合蛋白如通式(II)所示：
    Ab-L-TGF-βRII ECD  (II)
    其中TGF-βRII ECD为TGF-βRII胞外区的截短形式；
    Ab为PD-L1抗体或其抗原结合片段；
    L为连接序列。
  5. 根据权利要求4所述的用途，其特征在于，其中所述的连接序列为(G 4S)xG，其中x为3-6，优选为4。
  6. 根据权利要求5所述的用途，其特征在于，所述的TGF-βRII胞外区的包含如SEQ ID NO:11、12、13和14所示序列；优选包含SEQ ID NO:12所示的序列。
  7. 根据权利要求4所述的用途，其特征在于，所述的PD-L1抗体或其抗原结合片段包含：
    分别如SEQ ID NO:1、SEQ ID NO:2和SEQ ID NO:3所示的HCDR1、HCDR2和HCDR3，和分别如SEQ ID NO:4、SEQ ID NO:5和SEQ ID NO:6所示的LCDR1、LCDR2和LCDR3。
  8. 根据权利要求4-7任一项所述的用途，其特征在于，所述PD-L1抗体或其抗原结合片段为嵌合抗体、人源化抗体、全人抗体或其抗原结合片段。
  9. 根据权利要求8所述的用途，其特征在于，所述PD-L1抗体或其抗原结合片段包含如SEQ ID NO:7所示的重链可变区，和如SEQ ID NO:8所示的轻链可变区。
  10. 根据权利要求4-9任一项所述的用途，其特征在于，所述PD-L1抗体或其抗原结合片段的重链氨基酸序列如SEQ ID NO:9所示或与SEQ ID NO:9所示的氨基酸序列具有至少95％的同一性，轻链氨基酸序列如SEQ ID NO:10所示或与SEQ ID NO:10所示氨基酸序列具有至少95％的同一性。
  11. 根据权利要求4所述的用途，其特征在于，所述PD-L1抗体或其抗原结合片段选自avelumab、atezolizumab、durvalumab、CS-1001、M-7824、KL-A167、CX-072、、BGB-A333、GNS-1480、CA-170、BMS-936559，优选avelumab、atezolizumab、durvalumab。
  12. 根据权利要求4-10任一项所述的用途，其特征在于，所述TGF-β受体的融合蛋白中，PD-L1抗体或其抗原结合片段的重链氨基酸序列如SEQ ID NO:9所示，轻链氨基酸序列如SEQ ID NO:10所示，连接序列L为(G 4S) 4G，TGF-βRII胞外区的氨基酸序列如SEQ ID NO:12所示。
  13. 根据权利要求1或2所述的用途，其特征在于，所述多靶点酪氨酸激酶抑制剂为式(I)化合物或其可药用盐，
    Figure PCTCN2021114280-appb-100001
  14. 根据权利要求1-13任一项所述的用途，其特征在于，所述含有TGF-β受体的融合蛋白的给药剂量选自0.1-500mg/kg，给药频率为每周一次，每两周一次，每三周一次或每四周一次。
  15. 根据权利要求14所述的用途，其特征在于，所述含有TGF-β受体的融合蛋白的给药剂量选自10mg/kg、20mg/kg、30mg/kg、40mg/kg、50mg/kg、60mg/kg，给药频率为每两周一次，每三周一次或每四周一次；
    优选地，TGF-β受体的融合蛋白的给药剂量为30mg/kg，每三周一次。
  16. 根据权利要求1-13任一项所述的用途，其特征在于，所述含有TGF-β受体的融合蛋白的给药剂量选自1-4000mg，给药频率为每周一次，每两周一次，每三周一次或每四周一次优选的给药剂量选自300mg、600mg、900mg、1200mg、1500mg、1800mg、2100mg、2400mg、2700mg、3000mg、3300mg、3600mg。
  17. 根据权利要求1-16任一项所述的用途，其特征在于，所述多靶点酪氨酸 激酶抑制剂的给药剂量选自1-1600mg，给药频率为一日两次，一日一次，每两日一次，每三日一次，每四日一次，每五日一次，每六日一次，每周一次，每两周一次，每三周一次或每四周一次，优选一日一次或一日两次；
    优选地，多靶点酪氨酸激酶抑制剂的给药剂量为10mg、15mg、20mg、25mg、30mg，一日一次；
    更优选地，多靶点酪氨酸激酶抑制剂的给药剂量为20mg，一日一次。
  18. 根据权利要求17所述的用途，其特征在于，所述多靶点酪氨酸激酶抑制剂连续给药三周。
  19. 一种药物包装盒，其中含有根据权利要求1-18任一项所述的用途中所述的TGF-β受体的融合蛋白和所述多靶点酪氨酸激酶抑制剂。
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