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WO2025061119A1 - Anticorps anti-tigit et leurs utilisations - Google Patents

Anticorps anti-tigit et leurs utilisations Download PDF

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Publication number
WO2025061119A1
WO2025061119A1 PCT/CN2024/119890 CN2024119890W WO2025061119A1 WO 2025061119 A1 WO2025061119 A1 WO 2025061119A1 CN 2024119890 W CN2024119890 W CN 2024119890W WO 2025061119 A1 WO2025061119 A1 WO 2025061119A1
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seq
amino acid
antibody
set forth
cdrs
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Inventor
Weiqiu LAN
Baihong Liu
Yi Yang
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Biocytogen Pharmaceuticals Beijing Co Ltd
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Biocytogen Pharmaceuticals Beijing Co Ltd
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Publication of WO2025061119A1 publication Critical patent/WO2025061119A1/fr
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A61K39/39591—Stabilisation, fragmentation
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • C07K2317/732—Antibody-dependent cellular cytotoxicity [ADCC]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/94—Stability, e.g. half-life, pH, temperature or enzyme-resistance
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • This disclosure relates to anti-TIGIT antibodies or antigen binding fragments thereof, and antibody drug conjugates.
  • Cancer is currently one of the diseases that have the highest human mortality. According to the World Health Organization statistical data, in 2018, the number of global cancer incidence and death cases reached 18.1 million and 9.6 million, respectively. In China, the newly diagnosed cancer cases are 3.8 million, and the death toll is 2.3 million.
  • This disclosure relates to anti-TIGIT antibodies, antigen-binding fragment thereof, and the uses thereof.
  • the disclosure is related to an antibody or antigen-binding fragment thereof that binds to TIGIT (T cell immunoreceptor with Ig and ITIM domains) comprising:
  • VH heavy chain variable region
  • CDRs complementarity determining regions
  • VL light chain variable region
  • the VL CDR1 region comprises an amino acid sequence that is at least 80%identical to a selected VL CDR1 amino acid sequence
  • the VL CDR2 region comprises an amino acid sequence that is at least 80%identical to a selected VL CDR2 amino acid sequence
  • the VL CDR3 region comprises an amino acid sequence that is at least 80%identical to a selected VL CDR3 amino acid sequence
  • selected VH CDRs 1, 2, and 3 amino acid sequences and the selected VL CDRs, 1, 2, and 3 amino acid sequences are one of the following:
  • the selected VH CDRs 1, 2, 3 amino acid sequences are set forth in SEQ ID NOs: 43-45, respectively, and the selected VL CDRs 1, 2, 3 amino acid sequences are set forth in SEQ ID NOs: 28-30, respectively.
  • the VH comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 1-3, respectively, and the VL comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 4-6, respectively, according to the Kabat definition.
  • the VH comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 7-9, respectively, and the VL comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 10-12, respectively, according to the Kabat definition.
  • the VH comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 13-15, respectively, and the VL comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 16-18, respectively, according to the Kabat definition.
  • the VH comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 13, 61, 15, respectively
  • the VL comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 16-18, respectively, according to the Kabat definition.
  • the VH comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 13, 62, 15, respectively
  • the VL comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 16-18, respectively, according to the Kabat definition.
  • the VH comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 19-21, respectively, and the VL comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 22-24, respectively, according to the Kabat definition.
  • the VH comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 25-27, respectively, and the VL comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 28-30, respectively, according to the Kabat definition.
  • the VH comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 31-33, respectively, and the VL comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 4-6, respectively, according to the Chothia definition.
  • the VH comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 34-36, respectively, and the VL comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 10-12, respectively, according to the Chothia definition.
  • the VH comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 37-39, respectively, and the VL comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 16-18, respectively, according to the Chothia definition.
  • the VH comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 40-42, respectively, and the VL comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 22-24, respectively, according to the Chothia definition.
  • the VH comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 43-45, respectively, and the VL comprises CDRs 1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 28-30, respectively, according to the Chothia definition.
  • the disclosure is related to an antibody or antigen-binding fragment thereof that binds to TIGIT comprising a heavy chain variable region (VH) comprising an amino acid sequence that is at least 90%identical to a selected VH sequence, and a light chain variable region (VL) comprising an amino acid sequence that is at least 90%identical to a selected VL sequence, wherein the selected VH sequence and the selected VL sequence are one of the following:
  • the selected VH sequence is SEQ ID NO: 46, and the selected VL sequence is SEQ ID NO: 47;
  • the selected VH sequence is SEQ ID NO: 48 or 65, and the selected VL sequence is SEQ ID NO: 49;
  • the selected VH sequence is SEQ ID NO: 50, and the selected VL sequence is SEQ ID NO: 51;
  • the selected VH sequence is SEQ ID NO: 63
  • the selected VL sequence is SEQ ID NO: 51;
  • the selected VH sequence is SEQ ID NO: 64, and the selected VL sequence is SEQ ID NO: 51;
  • the selected VH sequence is SEQ ID NO: 52, and the selected VL sequence is SEQ ID NO: 53;
  • the selected VH sequence is SEQ ID NO: 54
  • the selected VL sequence is SEQ ID NO: 55.
  • the VH comprises the sequence of SEQ ID NO: 46 and the VL comprises the sequence of SEQ ID NO: 47.
  • the VH comprises the sequence of SEQ ID NO: 48 and the VL comprises the sequence of SEQ ID NO: 49.
  • the VH comprises the sequence of SEQ ID NO: 65 and the VL comprises the sequence of SEQ ID NO: 49.
  • the VH comprises the sequence of SEQ ID NO: 50 and the VL comprises the sequence of SEQ ID NO: 51.
  • the VH comprises the sequence of SEQ ID NO: 63 and the VL comprises the sequence of SEQ ID NO: 51.
  • the VH comprises the sequence of SEQ ID NO: 64 and the VL comprises the sequence of SEQ ID NO: 51.
  • the VH comprises the sequence of SEQ ID NO: 52 and the VL comprises the sequence of SEQ ID NO: 53.
  • the VH comprises the sequence of SEQ ID NO: 54 and the VL comprises the sequence of SEQ ID NO: 55.
  • the disclosure is related to an antibody or antigen-binding fragment thereof that binds to TIGIT comprising
  • VH heavy chain variable region
  • VH CDR2 heavy chain variable region
  • VL light chain variable region
  • the selected VH sequence is SEQ ID NO: 46, and the selected VL sequence is SEQ ID NO: 47;
  • the selected VH sequence is SEQ ID NO: 48 or 65, and the selected VL sequence is SEQ ID NO: 49;
  • the selected VH sequence is SEQ ID NO: 50, and the selected VL sequence is SEQ ID NO: 51;
  • the selected VH sequence is SEQ ID NO: 63
  • the selected VL sequence is SEQ ID NO: 51;
  • the selected VH sequence is SEQ ID NO: 64, and the selected VL sequence is SEQ ID NO: 51;
  • the selected VH sequence is SEQ ID NO: 52, and the selected VL sequence is SEQ ID NO: 53;
  • the selected VH sequence is SEQ ID NO: 54
  • the selected VL sequence is SEQ ID NO: 55.
  • the antibody or antigen-binding fragment thereof specifically binds to human or monkey TIGIT.
  • the antibody or antigen-binding fragment thereof is a human or humanized antibody or antigen-binding fragment thereof.
  • the antibody or antigen-binding fragment is a single-chain variable fragment (scFv) or a multispecific antibody (e.g., a bispecific antibody) .
  • scFv single-chain variable fragment
  • a multispecific antibody e.g., a bispecific antibody
  • the disclosure is related to an antibody or antigen-binding fragment thereof that cross-competes with the antibody or antigen-binding fragment thereof described herein.
  • the disclosure is related to a nucleic acid comprising a polynucleotide encoding a polypeptide comprising:
  • an immunoglobulin heavy chain or a fragment thereof comprising a heavy chain variable region (VH) comprising complementarity determining regions (CDRs) 1, 2, and 3 comprising the amino acid sequences set forth in SEQ ID NOs: 1-3, respectively, and wherein the VH, when paired with a light chain variable region (VL) comprising the amino acid sequence set forth in SEQ ID NO: 47, binds to TIGIT;
  • VH heavy chain variable region
  • CDRs complementarity determining regions
  • VL light chain variable region
  • CDRs complementarity determining regions
  • an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3 comprising the amino acid sequences set forth in SEQ ID NOs: 7-9, respectively, and wherein the VH, when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO: 49, binds to TIGIT;
  • an immunoglobulin light chain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3 comprising the amino acid sequences set forth in SEQ ID NOs: 10-12, respectively, and wherein the VL, when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO: 48 or 65, binds to TIGIT;
  • an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3 comprising the amino acid sequences set forth in SEQ ID NOs: 13-15, respectively, and wherein the VH, when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO: 51, binds to TIGIT;
  • an immunoglobulin light chain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3 comprising the amino acid sequences set forth in SEQ ID NOs: 16-18, respectively, and wherein the VL, when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO: 50, binds to TIGIT;
  • an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3 comprising the amino acid sequences set forth in SEQ ID NOs: 13, 61, 15, respectively, and wherein the VH, when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO: 51, binds to TIGIT;
  • an immunoglobulin light chain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3 comprising the amino acid sequences set forth in SEQ ID NOs: 16-18, respectively, and wherein the VL, when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO: 63, binds to TIGIT;
  • an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3 comprising the amino acid sequences set forth in SEQ ID NOs: 13, 62, 15, respectively, and wherein the VH, when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO: 51, binds to TIGIT;
  • an immunoglobulin light chain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3 comprising the amino acid sequences set forth in SEQ ID NOs: 16-18, respectively, and wherein the VL, when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO: 64, binds to TIGIT;
  • an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3 comprising the amino acid sequences set forth in SEQ ID NOs: 19-21, respectively, and wherein the VH, when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO: 53, binds to TIGIT;
  • an immunoglobulin light chain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3 comprising the amino acid sequences set forth in SEQ ID NOs: 22-24, respectively, and wherein the VL, when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO: 52, binds to TIGIT;
  • an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3 comprising the amino acid sequences set forth in SEQ ID NOs: 25-27, respectively, and wherein the VH, when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO: 55, binds to TIGIT;
  • an immunoglobulin light chain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3 comprising the amino acid sequences set forth in SEQ ID NOs: 28-30, respectively, and wherein the VL, when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO: 54, binds to TIGIT;
  • an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3 comprising the amino acid sequences set forth in SEQ ID NOs: 31-33, respectively, and wherein the VH, when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO: 47, binds to TIGIT;
  • an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3 comprising the amino acid sequences set forth in SEQ ID NOs: 34-36, respectively, and wherein the VH, when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO: 49, binds to TIGIT;
  • an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3 comprising the amino acid sequences set forth in SEQ ID NOs: 37-39 respectively, and wherein the VH, when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO: 51, binds to TIGIT;
  • an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3 comprising the amino acid sequences set forth in SEQ ID NOs: 40-42 respectively, and wherein the VH, when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO: 53, binds to TIGIT; and
  • an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3 comprising the amino acid sequences set forth in SEQ ID NOs: 43-45, respectively, and wherein the VH, when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO: 55, binds to TIGIT.
  • the VH when paired with a VL specifically binds to human or monkey TIGIT.
  • the immunoglobulin heavy chain or the fragment thereof is a human or humanized immunoglobulin heavy chain or a fragment thereof
  • the immunoglobulin light chain or the fragment thereof is a human or humanized immunoglobulin light chain or a fragment thereof.
  • the nucleic acid encodes a single-chain variable fragment (scFv) or a multi-specific antibody (e.g., a bispecific antibody) .
  • scFv single-chain variable fragment
  • a multi-specific antibody e.g., a bispecific antibody
  • the nucleic acid is cDNA.
  • the disclosure is related to a vector comprising one or more of the nucleic acids described herein.
  • the disclosure is related to a vector comprising two of the nucleic acids described herein, in some embodiments, the vector encodes the VH region and the VL region that together bind to TIGIT.
  • the disclosure is related to a pair of vectors, in some embodiments, each vector comprises one of the nucleic acids described herein, in some embodiments, together the pair of vectors encodes the VH region and the VL region that together bind to TIGIT.
  • the disclosure is related to a cell comprising the vector described herein.
  • the cell is a CHO cell.
  • the disclosure is related to a cell comprising one or more of the nucleic acids described herein.
  • the disclosure is related to a method of producing an antibody or antigen-binding fragment thereof, the method comprising
  • the disclosure is related to an antibody-drug conjugate (ADC) comprising a therapeutic agent covalently bound to the antibody or antigen-binding fragment thereof described herein.
  • the therapeutic agent is a cytotoxic or cytostatic agent.
  • the disclosure is related to a method of treating a subject having cancer, the method comprising administering a therapeutically effective amount of a composition comprising the antibody or antigen-binding fragment thereof described herein, or the antibody-drug conjugate described herein, to the subject.
  • the cancer is melanoma, lung cancer (e.g., non-small cell lung carcinoma (NSCLC) , small cell lung carcinoma (SCLC) ) , colon cancer, hepatocellular carcinoma (HCC) , gastric cancer, breast cancer, pancreatic cancer, glioblastoma, hematological malignancies, renal carcinoma or ovarian cancer.
  • lung cancer e.g., non-small cell lung carcinoma (NSCLC) , small cell lung carcinoma (SCLC)
  • HCC hepatocellular carcinoma
  • gastric cancer e.g., breast cancer, pancreatic cancer, glioblastoma, hematological malignancies, renal carcinoma or ovarian cancer.
  • the cancer is melanoma, non-small cell lung carcinoma (NSCLC) , colon cancer, hepatocellular carcinoma (HCC) , gastric cancer, glioblastoma, hematological malignancies, renal carcinoma or ovarian cancer.
  • NSCLC non-small cell lung carcinoma
  • HCC hepatocellular carcinoma
  • gastric cancer glioblastoma
  • hematological malignancies renal carcinoma or ovarian cancer.
  • the method further comprises administering a therapeutically effective amount of an anti-PD1 antibody, an anti-PDL1 antibody, an anti-PDL2 antibody, an anti-LAG-3 antibody, an anti-OX40 antibody, an anti-CTLA-4 antibody, an anti-GITR antibody, an anti-TIM-3 antibody, an anti-4-1BB antibody, and/or an anti-CD40 antibody, to the subject.
  • the method further comprises administering a chemotherapy to the subject.
  • the disclosure is related to a method of decreasing the rate of tumor growth, the method comprising contacting a tumor cell with an effective amount of a composition comprising the antibody or antigen-binding fragment thereof described herein, or the antibody-drug conjugate described herein.
  • the disclosure is related to a method of killing a tumor cell, the method comprising contacting a tumor cell with an effective amount of a composition comprising the antibody or antigen-binding fragment thereof described herein, or the antibody-drug conjugate described herein.
  • the disclosure is related to a pharmaceutical composition
  • a pharmaceutical composition comprising a pharmaceutically acceptable carrier and
  • malignancies of the various organ systems such as head and neck, respiratory, cardiovascular, renal, reproductive, hematological, neurological, hepatic, gastrointestinal, and endocrine systems; as well as adenocarcinomas which include malignancies such as most colon cancers, renal-cell carcinoma, prostate cancer and/or testicular tumors, non-small cell carcinoma of the lung, glioma and cancer of the small intestine.
  • Cancer that is “naturally arising” includes any cancer that is not experimentally induced by implantation of cancer cells into a subject, and includes, for example, spontaneously arising cancer, cancer caused by exposure of a patient to a carcinogen (s) , cancer resulting from insertion of a transgenic oncogene or knockout of a tumor suppressor gene, and cancer caused by infections, e.g., viral infections.
  • a carcinogen s
  • cancer resulting from insertion of a transgenic oncogene or knockout of a tumor suppressor gene and cancer caused by infections, e.g., viral infections.
  • the term “carcinoma” is art recognized and refers to malignancies of epithelial or endocrine tissues. The term also includes carcinosarcomas, which include malignant tumors composed of carcinomatous and sarcomatous tissues.
  • Adenocarcinoma refers to a carcinoma derived from glandular tissue or in which the tumor cells form recognizable glandular structures.
  • the term “sarcoma” is art recognized and refers to malignant tumors of mesenchymal derivation.
  • hematopoietic neoplastic disorders includes diseases involving hyperplastic/neoplastic cells of hematopoietic origin. Ahematopoietic neoplastic disorder can arise from myeloid, lymphoid or erythroid lineages, or precursor cells thereof.
  • a hematologic cancer is a cancer that begins in blood-forming tissue, such as the bone marrow, or in the cells of the immune system. Examples of hematologic cancer include e.g., leukemia, lymphoma, and multiple myeloma etc.
  • an antibody is used herein in its broadest sense and includes certain types of immunoglobulin molecules that include one or more antigen-binding domains that specifically bind to an antigen or epitope.
  • An antibody specifically includes, e.g., intact antibodies (e.g., intact immunoglobulins) , antibody fragments, bispecific antibodies, and multi-specific antibodies.
  • an antibody is a protein complex that includes two heavy chains and two light chains. Additional examples of an antibody are described herein.
  • the term “antigen-binding fragment” or “antibody fragment” refers to a portion of a full-length antibody, wherein the portion of the antibody is capable of specifically binding to an antigen.
  • the antigen-binding fragment contains at least one variable domain (e.g., a variable domain of a heavy chain or a variable domain of light chain or a VHH) .
  • variable domains include, e.g., Fab, Fab’, F (ab’) 2 , and Fv fragments.
  • human antibody refers to an antibody that is encoded by an endogenous nucleic acid (e.g., rearranged human immunoglobulin heavy or light chain locus) derived from a human.
  • a human antibody is collected from a human or produced in a human cell culture (e.g., human hybridoma cells) .
  • a human antibody is produced in a non-human cell (e.g., a mouse or hamster cell line) .
  • a human antibody is produced in a bacterial or yeast cell.
  • a human antibody is produced in a transgenic non-human animal (e.g., a bovine) containing an unrearranged or rearranged human immunoglobulin locus (e.g., heavy or light chain human immunoglobulin locus) .
  • a transgenic non-human animal e.g., a bovine
  • human immunoglobulin locus e.g., heavy or light chain human immunoglobulin locus
  • humanized antibody refers to a non-human antibody which contains minimal sequence derived from a non-human (e.g., mouse) immunoglobulin and contains sequences derived from a human immunoglobulin.
  • humanized antibodies are human antibodies (recipient antibody) in which hypervariable (e.g., CDR) region residues of the recipient antibody are replaced by hypervariable (e.g., CDR) region residues from a non-human antibody (e.g., a donor antibody) , e.g., a mouse, rat, or rabbit antibody, having the desired specificity, affinity, and capacity.
  • the Fv framework residues of the human immunoglobulin are replaced by corresponding non-human (e.g., mouse) immunoglobulin residues.
  • humanized antibodies may contain residues which are not found in the recipient antibody or in the donor antibody. These modifications can be made to further refine antibody performance.
  • the humanized antibody contains substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops (CDRs) correspond to those of a non-human (e.g., mouse) immunoglobulin and all or substantially all of the framework regions are those of a human immunoglobulin.
  • CDRs hypervariable loops
  • the humanized antibody can also contain at least a portion of an immunoglobulin constant region (Fc) , typically, that of a human immunoglobulin.
  • Fc immunoglobulin constant region
  • Humanized antibodies can be produced using molecular biology methods known in the art. Non-limiting examples of methods for generating humanized antibodies are described herein.
  • multispecific antibody refers to an antibody that binds to two or more different epitopes.
  • the epitopes can be on the same antigen or on different antigens.
  • the multispecific antibody is a bispecific antibody.
  • bispecific antibody refers to an antibody that binds to two different epitopes.
  • the epitopes can be on the same antigen or on different antigens.
  • FIG. 1 lists Kabat CDR sequences for anti-TIGIT antibodies.
  • FIG. 2 lists Chothia CDR sequences for anti-TIGIT antibodies.
  • FIG. 3 lists amino acids sequences discussed in the disclosure.
  • FIG. 4A shows the blocking effect of the binding between TIGIT and CD155 in the presence of an anti-TIGIT antibody (5F1 or 9H11) .
  • PC was used as a control.
  • FIG. 4B shows the blocking effect of the binding between TIGIT and CD155 in the presence of an anti-TIGIT antibody (36F3 or 36H7) .
  • PC was used as a control.
  • FIG. 4C shows the blocking effect of the binding between TIGIT and CD155 in the presence of an anti-TIGIT antibody 15C5.
  • PC was used as a control.
  • FIG. 5 shows the concentration-time curves of anti-TIGIT antibodies in different groups of C57BL/6 mice.
  • FIG. 6 shows the ADCC activity assay results of anti-TIGIT antibodies. PC was used as a control.
  • FIG. 7 shows the average tumor volume in different groups of hTIGIT mice that were injected with MC38 cells, and were treated with PC (G2) , 36F3 (G3) , 36H7 (G4) , or 9H11 (G5) treatment.
  • PC PC
  • G3 36F3
  • G4 36H7
  • G5 9H11
  • FIG. 8 shows the average tumor volume in different groups of hTIGIT mice that were injected with MC38 cells, and were treated with PC (G2) , 9H11 (G3, G4) or 36F3 (G5, G6) .
  • PC PC
  • 9H11 G3, G4
  • 36F3 G5, G6
  • PBS was used as a control (G1) .
  • FIG. 9 shows the average tumor volume in different groups of hTIGIT mice that were injected with MC38 cells, and were treated with PC (G2) or 15C5 (G3) .
  • PBS was used as a control (G1) .
  • the present disclosure relates to anti-TIGIT antibodies or antigen binding fragments thereof, and antibody drug conjugates.
  • TIGIT T cell immunoreceptor with Ig and ITIM domains, also called WUCAM, Vstm3, VSIG9
  • WUCAM WUCAM
  • Vstm3 VSIG9 T cell immunoreceptor with Ig and ITIM domains, also called WUCAM, Vstm3, VSIG9
  • TIGIT participates in a complex regulatory network involving multiple IRs (e.g., CD96/TACTILE, CD112R/PVRIG) , one competing costimulatory receptor (DNAM-1/CD226) , and multiple ligands (e.g., CD155 (PVR/NECL-5) , CD112 (Nectin-2/PVRL2) ) .
  • TIGIT -/- mice do not develop autoimmunity.
  • TIGIT -/- mice develop more severe experimental autoimmune encephalitis when immunized with myelin oligodendrocyte glycoprotein. Such an observation supports the role of TIGIT as a negative regulator of T cell functions.
  • TIGIT is expressed by activated CD8+T and CD4+T cells, natural killer (NK) cells, regulatory T cells (Tregs) , and follicular T helper cells in humans. In sharp contrast with DNAM-1/CD226, TIGIT is weakly expressed by naive T cells.
  • TIGIT is coexpressed with PD-1 on tumor antigen-specific CD8+T cells and CD8+tumor-infiltrating lymphocytes (TILs) in mice and humans. It is also coexpressed with other IRs, such as T cell immunoglobulin and mucin domain-containing molecule-3 (TIM-3) and lymphocyte activation gene 3 (LAG-3) , on exhausted CD8+T cell subsets in tumors. Further, TIGIT is highly expressed by Tregs in peripheral blood mononuclear cells of healthy donors and patients with cancer and further upregulated in the TME.
  • TIGIT is composed of an extracellular immunoglobulin (Ig) variable domain, a type 1 transmembrane domain, and a cytoplasmic tail with two inhibitory motifs conserved in mouse and human: an immunoreceptor tyrosine-based inhibitory motif (ITIM) and an Ig tail-tyrosine (ITT) -like motif.
  • ITIM immunoreceptor tyrosine-based inhibitory motif
  • ITT Ig tail-tyrosine
  • TIGIT in cancer immunotherapy.
  • TIGIT as an emerging immune checkpoint.
  • Targeting TIGIT for immunotherapy of cancer Update on clinical development.
  • the disclosure provides several antibodies and antigen-binding fragments thereof that specifically bind to TIGIT.
  • the antibodies and antigen-binding fragments described herein are capable of binding to TIGIT.
  • the disclosure provides e.g., anti-TIGIT antibodies 5F1, 9H11, 15C5, 36F3 and 36H7, and the antibodies derived therefrom (e.g., 15C5-ND, 15C5-SG) .
  • the CDR sequences for 5F1, and 5F1 derived antibodies include CDRs of the heavy chain variable domain, SEQ ID NOs: 1-3, and CDRs of the light chain variable domain, SEQ ID NOs: 4-6 as defined by Kabat definition.
  • the CDRs can also be defined by Chothia system. Under the Chothia definition, the CDR sequences of the heavy chain variable domain are set forth in SEQ ID NOs: 31-33, and CDR sequences of the light chain variable domain are set forth in SEQ ID NOs: 4-6.
  • the human heavy chain variable region and human light chain variable region for 5F1 are shown in SEQ ID NO: 46 and SEQ ID NO: 47, respectively.
  • the CDR sequences for 9H11, and 9H11 derived antibodies include CDRs of the heavy chain variable domain, SEQ ID NOs: 7-9, and CDRs of the light chain variable domain, SEQ ID NOs: 10-12, as defined by Kabat definition. Under Chothia definition, the CDR sequences of the heavy chain variable domain are set forth in SEQ ID NOs: 34-36, and CDRs of the light chain variable domain are set forth in SEQ ID NOs: 10-12.
  • the human heavy chain variable region and human light chain variable region for 9H11 are shown in SEQ ID NO: 65 and SEQ ID NO: 49, respectively.
  • the CDR sequences for 15C5, and 15C5 derived antibodies include CDRs of the heavy chain variable domain, SEQ ID NOs: 13-15, and CDRs of the light chain variable domain, SEQ ID NOs: 16-18, as defined by Kabat definition. Under Chothia definition, the CDR sequences of the heavy chain variable domain are set forth in SEQ ID NOs: 37-39, and CDRs of the light chain variable domain are set forth in SEQ ID NOs: 16-18.
  • the human heavy chain variable region and human light chain variable region for 15C5 are shown in SEQ ID NO: 50 and SEQ ID NO: 51, respectively.
  • the CDR sequences for 15C5 derived antibody 15C5-ND include CDRs of the heavy chain variable domain, SEQ ID NOs: 13, 61, 15, and CDRs of the light chain variable domain, SEQ ID NOs: 16-18, as defined by Kabat definition. Under Chothia definition, the CDR sequences of the heavy chain variable domain are set forth in SEQ ID NOs: 37-39, and CDRs of the light chain variable domain are set forth in SEQ ID NOs: 16-18.
  • the human heavy chain variable region and human light chain variable region for 15C5-ND are shown in SEQ ID NO: 63 and SEQ ID NO: 51, respectively.
  • the CDR sequences for 15C5 derived antibody 15C5-SG include CDRs of the heavy chain variable domain, SEQ ID NOs: 13, 62, 15, and CDRs of the light chain variable domain, SEQ ID NOs: 16-18, as defined by Kabat definition. Under Chothia definition, the CDR sequences of the heavy chain variable domain are set forth in SEQ ID NOs: 37-39, and CDRs of the light chain variable domain are set forth in SEQ ID NOs: 16-18.
  • the human heavy chain variable region and human light chain variable region for 15C5-SG are shown in SEQ ID NO: 64 and SEQ ID NO: 51, respectively.
  • the CDR sequences for 36F3, and 36F3 derived antibodies include CDRs of the heavy chain variable domain, SEQ ID NOs: 19-21, and CDRs of the light chain variable domain, SEQ ID NOs: 22-24, as defined by Kabat definition. Under Chothia definition, the CDR sequences of the heavy chain variable domain are set forth in SEQ ID NOs: 40-42, and CDRs of the light chain variable domain are set forth in SEQ ID NOs: 22-24.
  • the human heavy chain variable region and human light chain variable region for 36F3 are shown in SEQ ID NO: 52 and SEQ ID NO: 53, respectively.
  • the CDR sequences for 36H7, and 36H7 derived antibodies include CDRs of the heavy chain variable domain, SEQ ID NOs: 25-27, and CDRs of the light chain variable domain, SEQ ID NOs: 28-30, as defined by Kabat definition. Under Chothia definition, the CDR sequences of the heavy chain variable domain are set forth in SEQ ID NOs: 43-45, and CDRs of the light chain variable domain are set forth in SEQ ID NOs: 28-30.
  • the human heavy chain variable region and human light chain variable region for 36H7 are shown in SEQ ID NO: 54 and SEQ ID NO: 55, respectively.
  • the antibodies or antigen-binding fragments thereof described herein can also contain one, two, or three heavy chain variable region CDRs selected from the group of SEQ ID NOs: 1, 2, 3; SEQ ID NOs: 7, 8, 9; SEQ ID NOs: 13, 14, 15; SEQ ID NOs: 19, 20, 21; SEQ ID NOs: 25, 26, 27; SEQ ID NOs: 31, 32, 33; SEQ ID NOs: 34, 35, 36; SEQ ID NOs: 37, 38, 39; SEQ ID NOs: 40, 41, 42; and SEQ ID NOs: 43, 44, 45; and/or one, two, or three light chain variable region CDRs selected from the group of SEQ ID NOs: 4, 5, 6; SEQ ID NOs: 10, 11, 12; SEQ ID NOs: 16, 17, 18; SEQ ID NOs: 22, 23, 24; and SEQ ID NOs: 28, 29, 30.
  • the antibodies can have a heavy chain variable region (VH) comprising complementarity determining regions (CDRs) 1, 2, 3, wherein the CDR1 region comprises or consists of an amino acid sequence that is at least 80%, 85%, 90%, or 95%identical to a selected VH CDR1 amino acid sequence, the CDR2 region comprises or consists of an amino acid sequence that is at least 80%, 85%, 90%, or 95%identical to a selected VH CDR2 amino acid sequence, and the CDR3 region comprises or consists of an amino acid sequence that is at least 80%, 85%, 90%, or 95%identical to a selected VH CDR3 amino acid sequence, and a light chain variable region (VL) comprising CDRs 1, 2, 3, wherein the CDR1 region comprises or consists of an amino acid sequence that is at least 80%, 85%, 90%, or 95%identical to a selected VL CDR1 amino acid sequence, the CDR2 region comprises or consists of an amino acid sequence that is at least 80%
  • the antibody or an antigen-binding fragment described herein can contain a heavy chain variable domain containing one, two, or three of the CDRs of SEQ ID NO: 1 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 2 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 3 with zero, one or two amino acid insertions, deletions, or substitutions.
  • the antibody or an antigen-binding fragment described herein can contain a heavy chain variable domain containing one, two, or three of the CDRs of SEQ ID NO: 7 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 8 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 9 with zero, one or two amino acid insertions, deletions, or substitutions.
  • the antibody or an antigen-binding fragment described herein can contain a heavy chain variable domain containing one, two, or three of the CDRs of SEQ ID NO: 13 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 14 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 15 with zero, one or two amino acid insertions, deletions, or substitutions.
  • the antibody or an antigen-binding fragment described herein can contain a heavy chain variable domain containing one, two, or three of the CDRs of SEQ ID NO: 13 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 61 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 15 with zero, one or two amino acid insertions, deletions, or substitutions.
  • the antibody or an antigen-binding fragment described herein can contain a heavy chain variable domain containing one, two, or three of the CDRs of SEQ ID NO: 13 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 62 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 15 with zero, one or two amino acid insertions, deletions, or substitutions.
  • the antibody or an antigen-binding fragment described herein can contain a heavy chain variable domain containing one, two, or three of the CDRs of SEQ ID NO: 19 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 20 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 21 with zero, one or two amino acid insertions, deletions, or substitutions.
  • the antibody or an antigen-binding fragment described herein can contain a heavy chain variable domain containing one, two, or three of the CDRs of SEQ ID NO: 25 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 26 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 27 with zero, one or two amino acid insertions, deletions, or substitutions.
  • the antibody or an antigen-binding fragment described herein can contain a heavy chain variable domain containing one, two, or three of the CDRs of SEQ ID NO: 31 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 32 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 33 with zero, one or two amino acid insertions, deletions, or substitutions.
  • the antibody or an antigen-binding fragment described herein can contain a heavy chain variable domain containing one, two, or three of the CDRs of SEQ ID NO: 34 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 35 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 36 with zero, one or two amino acid insertions, deletions, or substitutions.
  • the antibody or an antigen-binding fragment described herein can contain a heavy chain variable domain containing one, two, or three of the CDRs of SEQ ID NO: 37 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 38 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 39 with zero, one or two amino acid insertions, deletions, or substitutions.
  • the antibody or an antigen-binding fragment described herein can contain a heavy chain variable domain containing one, two, or three of the CDRs of SEQ ID NO: 40 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 41 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 42 with zero, one or two amino acid insertions, deletions, or substitutions.
  • the antibody or an antigen-binding fragment described herein can contain a heavy chain variable domain containing one, two, or three of the CDRs of SEQ ID NO: 43 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 44 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 45 with zero, one or two amino acid insertions, deletions, or substitutions.
  • the antibody or an antigen-binding fragment described herein can contain a light chain variable domain containing one, two, or three o fthe CDRs of SEQ ID NO: 4 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 5 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 6 with zero, one or two amino acid insertions, deletions, or substitutions.
  • the antibody or an antigen-binding fragment described herein can contain a light chain variable domain containing one, two, or three o fthe CDRs of SEQ ID NO: 10 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 11 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 12 with zero, one or two amino acid insertions, deletions, or substitutions.
  • the antibody or an antigen-binding fragment described herein can contain a light chain variable domain containing one, two, or three of the CDRs of SEQ ID NO: 16 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 17 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 18 with zero, one or two amino acid insertions, deletions, or substitutions.
  • the antibody or an antigen-binding fragment described herein can contain a light chain variable domain containing one, two, or three of the CDRs of SEQ ID NO: 22 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 23 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 24 with zero, one or two amino acid insertions, deletions, or substitutions.
  • the antibody or an antigen-binding fragment described herein can contain a light chain variable domain containing one, two, or three of the CDRs of SEQ ID NO: 28 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 29 with zero, one or two amino acid insertions, deletions, or substitutions; SEQ ID NO: 30 with zero, one or two amino acid insertions, deletions, or substitutions.
  • the insertions, deletions, and substitutions can be within the CDR sequence, or at one or both terminal ends of the CDR sequence.
  • the CDR is determined based on Kabat definition scheme.
  • the CDR is determined based on Chothia definition scheme.
  • the CDR is determined based on a combination of Kabat and Chothia definition scheme.
  • the CDR is determined based on IMGT definition.
  • the disclosure also provides antibodies or antigen-binding fragments thereof that bind to TIGIT.
  • the antibodies or antigen-binding fragments thereof contain a heavy chain variable region (VH) comprising or consisting of an amino acid sequence that is at least 80%, 85%, 90%, or 95%identical to a selected VH sequence, and a light chain variable region (VL) comprising or consisting of an amino acid sequence that is at least 80%, 85%, 90%, or 95%identical to a selected VL sequence.
  • VH heavy chain variable region
  • VL light chain variable region
  • the selected VH sequence is SEQ ID NOs: 46, 48, 50, 52, 54, 63, 64, or 65
  • the selected VL sequence is SEQ ID NO: 47, 49, 51, 53, or 55.
  • the antibody or antigen binding fragment thereof can have 3 VH CDRs that are identical to the CDRs of any VH sequences as described herein. In some embodiments, the antibody or antigen binding fragment thereof can have 3VL CDRs that are identical to the CDRs of any VL sequences as described herein.
  • the disclosure also provides nucleic acid comprising a polynucleotide encoding a polypeptide comprising an immunoglobulin heavy chain or an immunoglobulin light chain.
  • the immunoglobulin heavy chain or immunoglobulin light chain comprises CDRs as shown in FIG. 1 or FIG. 2, or have sequences as shown in FIG. 3.
  • the polypeptides are paired with corresponding polypeptide (e.g., a corresponding heavy chain variable region or a corresponding light chain variable region)
  • TIGIT e.g., human TIGIT
  • the anti-TIGIT antibodies and antigen-binding fragments can also be antibody variants (including derivatives and conjugates) of antibodies or antibody fragments and multi-specific (e.g., bispecific) antibodies or antibody fragments.
  • Additional antibodies provided herein are polyclonal, monoclonal, multi-specific (multimeric, e.g., bispecific) , human antibodies, chimeric antibodies (e.g., human-mouse chimera) , single-chain antibodies, intracellularly-made antibodies (i.e., intrabodies) , and antigen-binding fragments thereof.
  • the antibodies or antigen-binding fragments thereof can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY) , class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2) , or subclass.
  • the antibody or antigen-binding fragment thereof is an IgG (e.g., IgG1) antibody or antigen-binding fragment thereof.
  • Fragments of antibodies are suitable for use in the methods provided so long as they retain the desired affinity and specificity of the full-length antibody.
  • a fragment of an antibody that binds to TIGIT will retain an ability to bind to TIGIT.
  • An Fv fragment is an antibody fragment which contains a complete antigen recognition and binding site. This region consists of a dimer of one heavy and one light chain variable domain in tight association, which can be covalent in nature, for example in scFv. It is in this configuration that the three CDRs of each variable domain interact to define an antigen binding site on the surface of the VH-VL dimer. Collectively, the six CDRs or a subset thereof confer antigen binding specificity to the antibody. However, even a single variable domain (or half of an Fv comprising only three CDRs specific for an antigen) can have the ability to recognize and bind antigen, although usually at a lower affinity than the entire binding site.
  • the present disclosure also provides an antibody or antigen-binding fragment thereof that cross-competes with any antibody or antigen-binding fragment as described herein.
  • the cross-competing assay is known in the art, and is described e.g., in Moore et al., "Antibody cross-competition analysis of the human immunodeficiency virus type 1 gp120 exterior envelope glycoprotein. " Journal of Virology 70.3 (1996) : 1863-1872, which is incorporated herein reference in its entirety.
  • the present disclosure also provides an antibody or antigen-binding fragment thereof that binds to the same epitope or region as any antibody or antigen-binding fragment as described herein.
  • the epitope binning assay is known in the art, and is described e.g., in Estep et al. "High throughput solution-based measurement of antibody-antigen affinity and epitope binning. " MAbs. Vol. 5. No. 2. Taylor&Francis, 2013, which is incorporated herein reference in its entirety.
  • antibodies can be made up of two classes of polypeptide chains, light chains and heavy chains.
  • a non-limiting antibody of the present disclosure can be an intact, four immunoglobulin chain antibody comprising two heavy chains and two light chains.
  • the heavy chain of the antibody can be of any isotype including IgM, IgG, IgE, IgA, or IgD or sub-isotype including IgG1, IgG2, IgG2a, IgG2b, IgG3, IgG4, IgE1, IgE2, etc.
  • the light chain can be a kappa light chain or a lambda light chain.
  • An antibody can comprise two identical copies of a light chain and/or two identical copies of a heavy chain.
  • the heavy chains which each contain one variable domain (or variable region, VH) and multiple constant domains (or constant regions) , bind to one another via disulfide bonding within their constant domains to form the “stem” of the antibody.
  • the light chains which each contain one variable domain (or variable region, VL) and one constant domain (or constant region) , each bind to one heavy chain via disulfide binding.
  • the variable region of each light chain is aligned with the variable region of the heavy chain to which it is bound.
  • the variable regions of both the light chains and heavy chains contain three hypervariable regions sandwiched between more conserved framework regions (FR) .
  • CDRs complementary determining regions
  • the four framework regions largely adopt a beta-sheet conformation and the CDRs form loops connecting, and in some cases forming part of, the beta-sheet structure.
  • the CDRs in each chain are held in close proximity by the framework regions and, with the CDRs from the other chain, contribute to the formation of the antigen-binding region.
  • the CDRs are important for recognizing an epitope of an antigen.
  • an “epitope” is the smallest portion of a target molecule capable of being specifically bound by the antigen binding domain of an antibody.
  • the minimal size of an epitope may be about three, four, five, six, or seven amino acids, but these amino acids need not be in a consecutive linear sequence of the antigen’s primary structure, as the epitope may depend on an antigen’s three-dimensional configuration based on the antigen’s secondary and tertiary structure.
  • the antibody is an intact immunoglobulin molecule (e.g., IgG1, IgG2a, IgG2b, IgG3, IgM, IgD, IgE, IgA) .
  • the IgG subclasses (IgG1, IgG2, IgG3, and IgG4) are highly conserved, differ in their constant region, particularly in their hinges and upper CH2 domains.
  • the sequences and differences of the IgG subclasses are known in the art, and are described, e.g., in Vidarsson, et al, “IgG subclasses and allotypes: from structure to effector functions. ” Frontiers in immunology 5 (2014) ; Irani, et al.
  • the antibody can also be an immunoglobulin molecule that is derived from any species (e.g., human, rodent, mouse, rat, camelid) .
  • Antibodies disclosed herein also include, but are not limited to, polyclonal, monoclonal, monospecific, polyspecific antibodies, and chimeric antibodies that include an immunoglobulin binding domain fused to another polypeptide.
  • the antigen binding domain or antigen binding fragment is a portion of an antibody that retains specific binding activity of the intact antibody, i.e., any portion of an antibody that is capable of specific binding to an epitope on the intact antibody’s target molecule. It includes, e.g., Fab, Fab’, F (ab’) 2 , and variants of these fragments.
  • an antibody or an antigen binding fragment thereof can be, e.g., a scFv, a Fv, a Fd, a dAb, a bispecific antibody, abispecific scFv, a diabody, a linear antibody, a single-chain antibody molecule, a multi-specific antibody formed from antibody fragments, and any polypeptide that includes a binding domain which is, or is homologous to, an antibody binding domain.
  • Non-limiting examples of antigen binding domains include, e.g., the heavy chain and/or light chain CDRs of an intact antibody, the heavy and/or light chain variable regions of an intact antibody, full length heavy or light chains of an intact antibody, or an individual CDR from either the heavy chain or the light chain of an intact antibody.
  • the Fab fragment contains a variable and constant domain of the light chain and a variable domain and the first constant domain (CH1) of the heavy chain.
  • F (ab') 2 antibody fragments comprise a pair of Fab fragments which are generally covalently linked near their carboxy termini by hinge cysteines between them. Other chemical couplings of antibody fragments are also known in the art.
  • Diabodies are small antibody fragments with two antigen-binding sites, which fragments comprise a VH connected to a VL in the same polypeptide chain (VH and VL) .
  • VH and VL polypeptide chain
  • Linear antibodies comprise a pair of tandem Fd segments (VH-CH1-VH-CH1) which, together with complementary light chain polypeptides, form a pair of antigen binding regions.
  • Linear antibodies can be bispecific or monospecific.
  • Antibodies and antibody fragments of the present disclosure can be modified in the Fc region to provide desired effector functions or serum half-life.
  • Multimerization of antibodies may be accomplished through natural aggregation of antibodies or through chemical or recombinant linking techniques known in the art. For example, some percentage of purified antibody preparations (e.g., purified IgG 1 molecules) spontaneously form protein aggregates containing antibody homodimers and other higher-order antibody multimers.
  • purified antibody preparations e.g., purified IgG 1 molecules
  • antibody homodimers may be formed through chemical linkage techniques known in the art.
  • heterobifunctional crosslinking agents including, but not limited to SMCC (succinimidyl4- (maleimidomethyl) cyclohexane-1-carboxylate) and SATA (N-succinimidyl S-acethylthio-acetate) can be used to form antibody multimers.
  • SMCC succinimidyl4- (maleimidomethyl) cyclohexane-1-carboxylate
  • SATA N-succinimidyl S-acethylthio-acetate
  • An exemplary protocol for the formation of antibody homodimers is described in Ghetie et al. (Proc. Natl. Acad. Sci. U.S.A. 94: 7509-7514, 1997) .
  • Antibody homodimers can be converted to Fab’ 2 homodimers through digestion with pepsin. Another way to form antibody homod
  • the multi-specific antibody is a bi-specific antibody.
  • Bi-specific antibodies can be made by engineering the interface between a pair of antibody molecules to maximize the percentage of heterodimers that are recovered from recombinant cell culture.
  • the interface can contain at least a part of the CH3 domain of an antibody constant domain.
  • one or more small amino acid side chains from the interface of the first antibody molecule are replaced with larger side chains (e.g., tyrosine or tryptophan) .
  • Compensatory “cavities” of identical or similar size to the large side chain (s) are created on the interface of the second antibody molecule by replacing large amino acid side chains with smaller ones (e.g., alanine or threonine) .
  • This provides a mechanism for increasing the yield of the heterodimer over other unwanted end-products such as homodimers. This method is described, e.g., in WO 96/27011, which is incorporated by reference in its entirety.
  • Bi-specific antibodies include cross-linked or “heteroconjugate” antibodies.
  • one of the antibodies in the heteroconjugate can be coupled to avidin and the other to biotin.
  • Heteroconjugate antibodies can also be made using any convenient cross-linking methods. Suitable cross-linking agents and cross-linking techniques are well known in the art and are disclosed in U.S. Patent No. 4,676,980, which is incorporated herein by reference in its entirety.
  • any of the antibodies or antigen-binding fragments described herein may be conjugated to a stabilizing molecule (e.g., a molecule that increases the half-life of the antibody or antigen-binding fragment thereof in a subject or in solution) .
  • stabilizing molecules include: a polymer (e.g., a polyethylene glycol) or a protein (e.g., serum albumin, such as human serum albumin) .
  • the conjugation of a stabilizing molecule can increase the half-life or extend the biological activity of an antibody or an antigen-binding fragment in vitro (e.g., in tissue culture or when stored as a pharmaceutical composition) or in vivo (e.g., in a human) .
  • the antigen binding fragment can form a part of a chimeric antigen receptor (CAR) .
  • the chimeric antigen receptor are fusions of single-chain variable fragments (scFv) as described herein, fused to CD3-zeta transmembrane-and endodomain.
  • the chimeric antigen receptor also comprises intracellular signaling domains from various costimulatory protein receptors (e.g., CD28, 41BB, ICOS) .
  • the chimeric antigen receptor comprises multiple signaling domains, e.g., CD3z-CD28-41BB or CD3z-CD28-OX40, to increase potency.
  • the disclosure further provides cells (e.g., T cells) that express the chimeric antigen receptors as described herein.
  • the antibodies or the antigen-binding fragments thereof described herein can be conjugated to a therapeutic agent.
  • the antibody-drug conjugate comprising the antibody or antigen-binding fragment thereof can covalently or non-covalently bind to a therapeutic agent.
  • the therapeutic agent is a cytotoxic or cytostatic agent (e.g., monomethyl auristatin E, monomethyl auristatin F, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicin, doxorubicin, daunorubicin, dihydroxy anthracin, maytansinoids such as DM-1 and DM-4, dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, puromycin, epirubicin, and cyclophosphamide and analogs) .
  • cytotoxic or cytostatic agent e.g., monomethyl auristatin E, monomethyl auristatin F, cytochalas
  • the therapeutic agent is MMAE or MMAF.
  • the therapeutic agent is conjugated via a linker, e.g., a VC linker. Details of the linkers used for ADCs can be found, e.g., in Su, Z. et al. “Antibody–drug conjugates: Recent advances in linker chemistry. ” Acta Pharmaceutica Sinica B (2021) , which is incorporated herein by reference in its entirety.
  • sequences e.g., CDRs or VH/VL sequences
  • an addition antigen e.g., OX40, CD40, 4-1BB, CTLA4, PD-1 or PD-L1 .
  • the disclosure provides antibodies and antigen-binding fragments thereof that specifically bind to TIGIT.
  • the antibodies and antigen-binding fragments described herein are capable of binding to TIGIT. These antibodies can be agonists or antagonists.
  • the anti-TIGIT antibodies or antigen-binding fragments thereof described herein can bind to TIGIT, and block the binding between TIGIT and CD155, and/or the binding between TIGIT and CD112. By binding to TIGIT, the anti-TIGIT antibodies can down-regulate or up-regulate the TIGIT-associated signaling pathway.
  • the anti-TIGIT antibodies or antigen-binding fragments thereof described herein can bind to TIGIT, but don't block the binding between TIGIT and CD155, and/or the binding between TIGIT and CD112.
  • the antibodies or antigen-binding fragments thereof as described herein are TIGIT agonist.
  • the antibodies or antigen-binding fragments thereof are TIGIT antagonist.
  • the antibodies or antigen-binding fragments thereof can bind to TIGIT (e.g., human TIGIT, monkey TIGIT, mouse TIGIT, and/or chimeric TIGIT) with a dissociation rate (koff) of less than 0.1 s -1 , less than 0.01 s -1 , less than 0.001 s -1 , less than 0.0001 s -1 , or less than 0.00001 s -1 .
  • the dissociation rate (koff) is greater than 0.01 s -1 , greater than 0.001 s -1 , greater than 0.0001 s -1 , greater than 0.00001 s -1 , or greater than 0.000001 s -1 .
  • kinetic association rates (kon) is greater than 1 x 10 2 /Ms, greater than 1 x 10 3 /Ms, greater than 1 x 10 4 /Ms, greater than 1 x 10 5 /Ms, or greater than 1 x 10 6 /Ms. In some embodiments, kinetic association rates (kon) is less than 1 x 10 5 /Ms, less than 1 x 10 6 /Ms, or less than 1 x 10 7 /Ms.
  • the antibodies or antigen-binding fragments thereof can bind to TIGIT (e.g., human TIGIT, monkey TIGIT, mouse TIGIT, and/or chimeric TIGIT) with a KD of less than 1 x 10 -6 M, less than 1 x 10 -7 M, less than 1 x 10 -8 M, less than 1 x 10 -9 M, or less than 1 x 10 -10 M.
  • TIGIT e.g., human TIGIT, monkey TIGIT, mouse TIGIT, and/or chimeric TIGIT
  • the KD is less than 50 nM, 40 nM, 30 nM, 20 nM, 10 nM, 9 nM, 8 nM, 7 nM, 6 nM, 5 nM, 4 nM, 3 nM, 2 nM, or 1 nM. In some embodiments, KD is greater than 1 x 10 -7 M, greater than 1 x 10 -8 M, greater than 1 x 10 -9 M, or greater than 1 x 10 -10 M.
  • the antibodies or the antigen-binding fragments thereof have a tumor growth inhibition rate or percentage (TGI%) that is greater than 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190%, or 200%.
  • the antibody has a tumor growth inhibition percentage that is less than 60%, 70%, 80%, 90%, 100%, 110%, 120%, 130%, 140%, or 150%.
  • the TGI (%) can be determined, e.g., at 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 days after the treatment starts.
  • Ti is the average tumor volume in the treatment group on day i.
  • T0 is the average tumor volume in the treatment group on day zero.
  • Vi is the average tumor volume in the control group on day i.
  • V0 is the average tumor volume in the control group on day zero.
  • the antibody or the antigen-binding fragment thereof can enhance the amount of tumor-infiltrating lymphocytes. In some embodiments, the antibody or the antigen-binding fragment thereof can enhance the amount of tumor-infiltrating lymphocytes by greater than 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190%, or 200%.
  • the antibody or the antigen-binding fragment thereof has a functional Fc region.
  • effector function of a functional Fc region is antibody-dependent cell-mediated cytotoxicity (ADCC) .
  • ADCC antibody-dependent cell-mediated cytotoxicity
  • effector function of a functional Fc region is phagocytosis.
  • effector function of a functional Fc region is ADCC and phagocytosis.
  • the Fc region is human IgG1, human IgG2, human IgG3, or human IgG4.
  • the antibody or the antigen-binding fragment thereof does not have a functional Fc region.
  • the antibodies or antigen binding fragments are Fab, Fab’, F (ab’) 2 , and Fv fragments.
  • the antibody or the antigen-binding fragment thereof as described herein have an Fc region without effector function.
  • the Fc is a human IgG4 Fc.
  • the Fc does not have a functional Fc region.
  • the Fc region has LALA mutations (L234A and L235A mutations in EU numbering) , or LALA-PG mutations (L234A, L235A, P329G mutations in EU numbering) .
  • Fc region a cysteine residue (s) can be introduced into the Fc region, thereby allowing interchain disulfide bond formation in this region.
  • the homodimeric fusion protein thus generated may have any increased half-life in vitro and/or in vivo.
  • Fc regions are provided having a carbohydrate structure that lacks fucose attached (directly or indirectly) to an Fc region.
  • the amount of fucose in such Fc region composition may be from 1%to 80%, from 1%to 65%, from 5%to 65%or from 20%to 40%.
  • the amount of fucose is determined by calculating the average amount of fucose within the sugar chain at Asn 297, relative to the sum of all glycostructures attached to Asn 297 (e.g. complex, hybrid and high mannose structures) as measured by MALDI-TOF mass spectrometry, as described in WO 2008/077546, for example.
  • Asn297 refers to the asparagine residue located at about position 297 in the Fc region (Eu numbering of Fc region residues; or position 314 in Kabat numbering) ; however, Asn297 may also be located about ⁇ 3 amino acids upstream or downstream of position 297, i.e., between positions 294 and 300, due to minor sequence variations in Fc region sequences. Such fucosylation variants may have improved ADCC function.
  • the Fc region can be further engineered to replace the Asparagine at position 297 with Alanine (N297A) .
  • An isolated fragment of human protein can be used as an immunogen to generate antibodies using standard techniques for polyclonal and monoclonal antibody preparation.
  • Polyclonal antibodies can be raised in animals by multiple injections (e.g., subcutaneous or intraperitoneal injections) of an antigenic peptide or protein.
  • the antigenic peptide or protein is injected with at least one adjuvant.
  • the antigenic peptide or protein can be conjugated to an agent that is immunogenic in the species to be immunized. Animals can be injected with the antigenic peptide or protein more than one time (e.g., twice, three times, four times, or five times) .
  • the full-length polypeptide or protein can be used or, alternatively, antigenic peptide fragments thereof can be used as immunogens.
  • the antigenic peptide of a protein comprises at least 8 (e.g., at least 10, 15, 20, or 30) amino acid residues of the amino acid sequence of the protein and encompasses an epitope of the protein such that an antibody raised against the peptide forms a specific immune complex with the protein.
  • An immunogen typically is used to prepare antibodies by immunizing a suitable subject (e.g., human or transgenic animal expressing at least one human immunoglobulin locus) .
  • a suitable subject e.g., human or transgenic animal expressing at least one human immunoglobulin locus
  • An appropriate immunogenic preparation can contain, for example, a recombinantly-expressed or a chemically-synthesized polypeptide.
  • the preparation can further include an adjuvant, such as Freund’s complete or incomplete adjuvant, or a similar immunostimulatory agent.
  • Polyclonal antibodies can be prepared as described above by immunizing a suitable subject with a polypeptide, or an antigenic peptide thereof (e.g., part of the protein) as an immunogen.
  • the antibody titer in the immunized subject can be monitored over time by standard techniques, such as with an enzyme-linked immunosorbent assay (ELISA) using the immobilized polypeptide or peptide.
  • ELISA enzyme-linked immunosorbent assay
  • the antibody molecules can be isolated from the mammal (e.g., from the blood) and further purified by well-known techniques, such as protein A or protein G chromatography to obtain the IgG fraction.
  • antibody-producing cells can be obtained from the subject and used to prepare monoclonal antibodies by standard techniques, such as the hybridoma technique originally described by Kohler et al. (Nature 256: 495-497, 1975) , the human B cell hybridoma technique (Kozbor et al., Immunol. Today 4: 72, 1983) , the EBV-hybridoma technique (Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc., pp. 77-96, 1985) , or trioma techniques.
  • standard techniques such as the hybridoma technique originally described by Kohler et al. (Nature 256: 495-497, 1975) , the human B cell hybridoma technique (Kozbor et al., Immunol. Today 4: 72, 1983) , the EBV-hybridoma technique (Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Lis
  • Hybridoma cells producing a monoclonal antibody are detected by screening the hybridoma culture supernatants for antibodies that bind the polypeptide or epitope of interest, e.g., using a standard ELISA assay.
  • Variants of the antibodies or antigen-binding fragments described herein can be prepared by introducing appropriate nucleotide changes into the DNA encoding a human, humanized, or chimeric antibody, or antigen-binding fragment thereof described herein, or by peptide synthesis.
  • Such variants include, for example, deletions, insertions, or substitutions of residues within the amino acids sequences that make-up the antigen-binding site of the antibody or an antigen-binding domain.
  • some antibodies or antigen-binding fragments will have increased affinity for the target protein. Any combination of deletions, insertions, and/or combinations can be made to arrive at an antibody or antigen-binding fragment thereof that has increased binding affinity for the target.
  • the amino acid changes introduced into the antibody or antigen-binding fragment can also alter or introduce new post-translational modifications into the antibody or antigen-binding fragment, such as changing (e.g., increasing or decreasing) the number of glycosylation sites, changing the type of glycosylation site (e.g., changing the amino acid sequence such that a different sugar is attached by enzymes present in a cell) , or introducing new glycosylation sites.
  • Antibodies disclosed herein can be derived from any species of animal, including mammals.
  • Non-limiting examples of native antibodies include antibodies derived from humans, primates, e.g., monkeys and apes, cows, pigs, horses, sheep, camelids (e.g., camels and llamas) , chicken, goats, and rodents (e.g., rats, mice, hamsters and rabbits) , including transgenic rodents genetically engineered to produce human antibodies.
  • Phage display can be used to optimize antibody sequences with desired binding affinities.
  • a gene encoding single chain Fv (comprising VH or VL) can be inserted into a phage coat protein gene, causing the phage to “display” the scFv on its outside while containing the gene for the protein on its inside, resulting in a connection between genotype and phenotype.
  • These displaying phages can then be screened against target antigens, in order to detect interaction between the displayed antigen binding sites and the target antigen.
  • large libraries of proteins can be screened and amplified in a process called in vitro selection, and antibodies sequences with desired binding affinities can be obtained.
  • Human and humanized antibodies include antibodies having variable and constant regions derived from (or having the same amino acid sequence as those derived from) human germline immunoglobulin sequences. Human antibodies may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo) , for example in the CDRs.
  • a humanized antibody typically has a human framework (FR) grafted with non-human CDRs.
  • FR human framework
  • a humanized antibody has one or more amino acid sequence introduced into it from a source which is non-human.
  • “humanized” antibodies are chimeric antibodies wherein substantially less than an intact human V domain has been substituted by the corresponding sequence from a non-human species.
  • humanized antibodies are typically mouse antibodies in which some CDR residues and some FR residues are substituted by residues from analogous sites in human antibodies.
  • humanized antibodies can be prepared by a process of analysis of the parental sequences and various conceptual humanized products using three-dimensional models of the parental and humanized sequences.
  • Three-dimensional immunoglobulin models are commonly available and are familiar to those skilled in the art.
  • Computer programs are available which illustrate and display probable three-dimensional conformational structures of selected candidate immunoglobulin sequences. Inspection of these displays permits analysis of the likely role of the residues in the functioning of the candidate immunoglobulin sequence, i.e., the analysis of residues that influence the ability of the candidate immunoglobulin to bind its antigen.
  • FR residues can be selected and combined from the recipient and import sequences so that the desired antibody characteristic, such as increased affinity for the target antigen (s) , is achieved.
  • a mouse e.g., RenMab TM mouse with a humanized heavy chain immunoglobulin locus and a humanized kappa chain immunoglobulin locus is used to generate antibodies.
  • the heavy chain immunoglobulin locus is a region on the chromosome that contains genes for the heavy chains of antibodies.
  • the locus can include e.g., human IGHV (variable) genes, human IGHD (diversity) genes, human IGHJ (joining) genes, and mouse heavy chain constant domain genes.
  • the kappa chain immunoglobulin locus is a region on the chromosome that contains genes that encode the light chains of antibodies (kappa chain) .
  • the kappa chain immunoglobulin locus can include e.g., human IGKV (variable) genes, human IGKJ (joining) genes, and mouse light chain constant domain genes.
  • human IGKV variable
  • human IGKJ joining
  • mouse light chain constant domain genes e.g., RenMab TM mice.
  • RenMab TM mice can be found in PCT/CN2020/075698 or US20200390073A1, which is incorporated herein by reference in its entirety.
  • Identity or homology with respect to an original sequence is usually the percentage of amino acid residues present within the candidate sequence that are identical with a sequence present within the human, humanized, or chimeric antibody or fragment, after aligning the sequences and introducing gaps, ifnecessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity.
  • a covalent modification can be made to the antibodies or the antigen-binding fragments thereof. These covalent modifications can be made by chemical or enzymatic synthesis, or by enzymatic or chemical cleavage. Other types of covalent modifications of the antibody or antibody fragment are introduced into the molecule by reacting targeted amino acid residues of the antibody or fragment with an organic derivatization agent that is capable of reacting with selected side chains or the N-or C-terminal residues.
  • antibody variants having a carbohydrate structure that lacks fucose attached (directly or indirectly) to an Fc region.
  • the amount of fucose in such antibody may be from 1%to 80%, from 1%to 65%, from 5%to 65%or from 20%to 40%.
  • the amount of fucose is determined by calculating the average amount of fucose within the sugar chain at Asn297, relative to the sum of all glycostructures attached to Asn 297 (e.g. complex, hybrid and high mannose structures) as measured by MALDI-TOF mass spectrometry, as described in WO 2008/077546, for example.
  • Asn297 refers to the asparagine residue located at about position 297 in the Fc region (Eu numbering of Fc region residues; or position 314 in Kabat numbering) ; however, Asn297 may also be located about ⁇ 3 amino acids upstream or downstream of position 297, i.e., between positions 294 and 300, due to minor sequence variations in antibodies. Such fucosylation variants may have improved ADCC function.
  • the Fc region of the antibody can be further engineered to replace the Asparagine at position 297 with Alanine (N297A) .
  • the Fc region of the antibodies was further engineered to replace the serine at position 228 (EU numbering) of IgG4 with proline (S228P) .
  • S228P serine at position 228
  • a detailed description regarding S228 mutation is described, e.g., in Silva et al. “The S228P mutation prevents in vivo and in vitro IgG4 Fab-arm exchange as demonstrated using a combination of novel quantitative immunoassays and physiological matrix preparation. ” Journal of Biological Chemistry 290.9 (2015) : 5462-5469, which is incorporated by reference in its entirety.
  • the present disclosure also provides recombinant vectors (e.g., expression vectors) that include an isolated polynucleotide disclosed herein (e.g., a polynucleotide that encodes a polypeptide disclosed herein) , host cells into which are introduced the recombinant vectors (i.e., such that the host cells contain the polynucleotide and/or a vector comprising the polynucleotide) , and the production of recombinant antibody polypeptides or fragments thereof by recombinant techniques.
  • recombinant vectors e.g., expression vectors
  • an isolated polynucleotide disclosed herein e.g., a polynucleotide that encodes a polypeptide disclosed herein
  • host cells into which are introduced the recombinant vectors (i.e., such that the host cells contain the polynucleotide and/or a vector comprising the polynucleotide
  • a “vector” is any construct capable of delivering one or more polynucleotide (s) of interest to a host cell when the vector is introduced to the host cell.
  • An “expression vector” is capable of delivering and expressing the one or more polynucleotide (s) of interest as an encoded polypeptide in a host cell into which the expression vector has been introduced.
  • the polynucleotide of interest is positioned for expression in the vector by being operably linked with regulatory elements such as a promoter, enhancer, and/or a poly-A tail, either within the vector or in the genome of the host cell at or near or flanking the integration site of the polynucleotide of interest such that the polynucleotide of interest will be translated in the host cell introduced with the expression vector.
  • regulatory elements such as a promoter, enhancer, and/or a poly-A tail
  • a vector can be introduced into the host cell by methods known in the art, e.g., electroporation, chemical transfection (e.g., DEAE-dextran) , transformation, transfection, and infection and/or transduction (e.g., with recombinant virus) .
  • vectors include viral vectors (which can be used to generate recombinant virus) , naked DNA or RNA, plasmids, cosmids, phage vectors, and DNA or RNA expression vectors associated with cationic condensing agents.
  • a polynucleotide disclosed herein e.g., a polynucleotide that encodes a polypeptide disclosed herein
  • a viral expression system e.g., vaccinia or other pox virus, retrovirus, or adenovirus
  • vaccinia or other pox virus, retrovirus, or adenovirus may involve the use of a non-pathogenic (defective) , replication competent virus, or may use a replication defective virus.
  • viral propagation generally will occur only in complementing virus packaging cells. Suitable systems are disclosed, for example, in Fisher-Hoch et al., 1989, Proc. Natl. Acad. Sci. USA 86: 317-321; Flexner et al., 1989, Ann. N.Y.
  • the DNA insert comprising an antibody-encoding or polypeptide-encoding polynucleotide disclosed herein can be operatively linked to an appropriate promoter (e.g., a heterologous promoter) , such as the phage lambda PL promoter, the E. coli lac, trp and tac promoters, the SV40 early and late promoters and promoters of retroviral LTRs, to name a few. Other suitable promoters are known to the skilled artisan.
  • the expression constructs can further contain sites for transcription initiation, termination and, in the transcribed region, a ribosome binding site for translation.
  • the coding portion of the mature transcripts expressed by the constructs may include a translation initiating at the beginning and a termination codon (UAA, UGA, or UAG) appropriately positioned at the end of the polypeptide to be translated.
  • the expression vectors can include at least one selectable marker.
  • markers include dihydrofolate reductase or neomycin resistance for eukaryotic cell culture and tetracycline or ampicillin resistance genes for culturing in E. coli and other bacteria.
  • Representative examples of appropriate hosts include, but are not limited to, bacterial cells, such as E. coli, Streptomyces, and Salmonella typhimurium cells; fungal cells, such as yeast cells; insect cells such as Drosophila S2 and Spodoptera Sf9 cells; animal cells such as CHO, COS, Bowes melanoma, and HK 293 cells; and plant cells. Appropriate culture mediums and conditions for the host cells described herein are known in the art.
  • Non-limiting vectors for use in bacteria include pQE70, pQE60 and pQE-9, available from Qiagen; pBS vectors, Phagescript vectors, Bluescript vectors, pNH8A, pNH16a, pNH18A, pNH46A, available from Stratagene; and ptrc99a, pKK223-3, pKK233-3, pDR540, pRIT5 available from Pharmacia.
  • Non-limiting eukaryotic vectors include pWLNEO, pSV2CAT, pOG44, pXT1 and pSG available from Stratagene; and pSVK3, pBPV, pMSG and pSVL available from Pharmacia. Other suitable vectors will be readily apparent to the skilled artisan.
  • Non-limiting bacterial promoters suitable for use include the E. coli lacI and lacZ promoters, the T3 and T7 promoters, the gpt promoter, the lambda PR and PL promoters and the trp promoter.
  • Suitable eukaryotic promoters include the CMV immediate early promoter, the HSV thymidine kinase promoter, the early and late SV40 promoters, the promoters of retroviral LTRs, such as those of the Rous sarcoma virus (RSV) , and metallothionein promoters, such as the mouse metallothionein-I promoter.
  • yeast Saccharomyces cerevisiae a number of vectors containing constitutive or inducible promoters such as alpha factor, alcohol oxidase, and PGH may be used.
  • constitutive or inducible promoters such as alpha factor, alcohol oxidase, and PGH.
  • Introduction of the construct into the host cell can be effected by calcium phosphate transfection, DEAE-dextran mediated transfection, cationic lipid-mediated transfection, electroporation, transduction, infection or other methods.
  • Such methods are described in many standard laboratory manuals, such as Davis et al., Basic Methods In Molecular Biology (1986) , which is incorporated herein by reference in its entirety.
  • Enhancers are cis-acting elements of DNA, usually about from 10 to 300bp that act to increase transcriptional activity of a promoter in a given host cell-type.
  • enhancers include the SV40 enhancer, which is located on the late side of the replication origin at base pairs 100 to 270, the cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, and adenovirus enhancers.
  • secretion signals may be incorporated into the expressed polypeptide.
  • the signals may be endogenous to the polypeptide or they may be heterologous signals.
  • the polypeptide (e.g., antibody) can be expressed in a modified form, such as a fusion protein (e.g., a GST-fusion) or with a histidine-tag, and may include not only secretion signals, but also additional heterologous functional regions. For instance, a region of additional amino acids, particularly charged amino acids, may be added to the N-terminus of the polypeptide to improve stability and persistence in the host cell, during purification, or during subsequent handling and storage. Also, peptide moieties can be added to the polypeptide to facilitate purification. Such regions can be removed prior to final preparation of the polypeptide. The addition of peptide moieties to polypeptides to engender secretion or excretion, to improve stability and to facilitate purification, among others, are familiar and routine techniques in the art.
  • the disclosure also provides a nucleic acid sequence that is at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%identical to any nucleotide sequence as described herein, and an amino acid sequence that is at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%identical to any amino acid sequence as described herein.
  • the disclosure also provides a nucleic acid sequence that has a homology of at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%to any nucleotide sequence as described herein, and an amino acid sequence that has a homology of at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%to any amino acid sequence as described herein.
  • the amino acid sequence is less than 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 250, 300, 350, 400, 500, 600, 700 or 800 amino acid residues.
  • the amino acid sequence (i) comprises an amino acid sequence; or (ii) consists of an amino acid sequence, wherein the amino acid sequence is any one of the sequences as described herein.
  • the nucleic acid sequence (i) comprises a nucleic acid sequence; or (ii) consists of a nucleic acid sequence, wherein the nucleic acid sequence is any one of the sequences as described herein.
  • the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or nucleic acid sequence for optimal alignment and non-homologous sequences can be disregarded for comparison purposes) .
  • the amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position (as used herein amino acid or nucleic acid “identity” is equivalent to amino acid or nucleic acid “homology” ) .
  • the percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences.
  • the comparison of sequences and determination of percent identity between two sequences can be accomplished using a Blossum 62 scoring matrix with a gap penalty of 12, a gap extend penalty of 4, and a frameshift gap penalty of 5.
  • the percentage of sequence homology (e.g., amino acid sequence homology or nucleic acid homology) can also be determined. How to determine percentage of sequence homology is known in the art.
  • amino acid residues conserved with similar physicochemical properties e.g. leucine and isoleucine, can be used to measure sequence similarity. Families of amino acid residues having similar physicochemical properties have been defined in the art.
  • amino acids with basic side chains e.g., lysine, arginine, histidine
  • acidic side chains e.g., aspartic acid, glutamic acid
  • uncharged polar side chains e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine
  • nonpolar side chains e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan
  • beta-branched side chains e.g., threonine, valine, isoleucine
  • aromatic side chains e.g., tyrosine, phenylalanine, tryptophan, histidine
  • the disclosure provides one or more nucleic acid encoding any of the polypeptides as described herein.
  • the nucleic acid e.g., cDNA
  • the nucleic acid includes a polynucleotide encoding a polypeptide of a heavy chain as described herein.
  • the nucleic acid includes a polynucleotide encoding a polypeptide of a light chain as described herein.
  • the nucleic acid includes a polynucleotide encoding a scFv polypeptide as described herein.
  • the vector can have two of the nucleic acids as described herein, wherein the vector encodes the VL region and the VH region that together bind to TIGIT.
  • a pair of vectors is provided, wherein each vector comprises one of the nucleic acids as described herein, wherein together the pair of vectors encodes the VL region and the VH region that together bind to TIGIT.
  • Vectors can also be constructed to express specific antibodies or polypeptides.
  • a vector can be constructed to co-express anti-TIGIT antibody light chain (TIGIT-K) and heavy chain (TIGIT-H) .
  • a vector can contain sequences of, from 5’ end to 3’ end, cytomegalovirus promotor (CMV) , TIGIT-K, polyadenylation (PolyA) , CMV, TIGIT-H, PolyA, simian vacuolating virus 40 terminator (SV40) and glutamine synthetase marker (GS) .
  • CMV cytomegalovirus promotor
  • TIGIT-K TIGIT-K
  • PolyA polyadenylation
  • CMV CMV
  • TIGIT-H polyadenylation
  • PolyA simian vacuolating virus 40 terminator
  • GS glutamine synthetase marker
  • a first vector expressing antibody heavy chains e.g., any of the heavy chains described herein
  • a second vector expressing antibody light chains e.g., any of the light chains described herein
  • co-transfect cells e.g., CHO cells
  • the methods described herein include methods for the treatment of disorders associated with cancer. Generally, the methods include administering a therapeutically effective amount of engineered antibodies or antigen-binding fragments thereof as described herein, to a subject who is in need of, or who has been determined to be in need of, such treatment.
  • to “treat” means to ameliorate at least one symptom of the disorder associated with cancer.
  • cancer results in death; thus, a treatment can result in an increased life expectancy (e.g., by at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months, or by at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 years) .
  • Administration of a therapeutically effective amount of an agent described herein for the treatment of a condition associated with cancer will result in decreased number of cancer cells and/or alleviated symptoms.
  • cancer refers to cells having the capacity for autonomous growth, i.e., an abnormal state or condition characterized by rapidly proliferating cell growth.
  • the term is meant to include all types of cancerous growths or oncogenic processes, metastatic tissues or malignantly transformed cells, tissues, or organs, irrespective of histopathologic type or stage of invasiveness.
  • tumor refers to cancerous cells, e.g., a mass of cancerous cells.
  • Cancers that can be treated or diagnosed using the methods described herein include malignancies of the various organ systems, such as affecting lung, breast, thyroid, lymphoid, gastrointestinal, and genito-urinary tract, as well as adenocarcinomas which include malignancies such as most colon cancers, renal-cell carcinoma, prostate cancer and/or testicular tumors, non-small cell carcinoma of the lung, cancer of the small intestine and cancer of the esophagus.
  • the agents described herein are designed for treating or diagnosing a carcinoma in a subject.
  • carcinoma is art recognized and refers to malignancies of epithelial or endocrine tissues including respiratory system carcinomas, gastrointestinal system carcinomas, genitourinary system carcinomas, testicular carcinomas, breast carcinomas, prostatic carcinomas, endocrine system carcinomas, and melanomas.
  • the cancer is renal carcinoma or melanoma.
  • Exemplary carcinomas include those forming from tissue of the cervix, lung, prostate, breast, head and neck, colon and ovary.
  • carcinosarcomas e.g., which include malignant tumors composed of carcinomatous and sarcomatous tissues.
  • an “adenocarcinoma” refers to a carcinoma derived from glandular tissue or in which the tumor cells form recognizable glandular structures.
  • the term “sarcoma” is art recognized and refers to malignant tumors of mesenchymal derivation.
  • the cancer is lung cancer, skin cancer, head and neck cancer, bladder cancer, prostate cancer, breast cancer, endometrium cancer, cervix cancer, pancreas cancer, glioma, lymphoma, or leukemia.
  • the cancer is melanoma, non-small cell lung carcinoma (NSCLC) , colon cancer, hepatocellular carcinoma (HCC) , gastric cancer, glioblastoma, hematological malignancies, renal carcinoma or ovarian cancer.
  • NSCLC non-small cell lung carcinoma
  • HCC hepatocellular carcinoma
  • gastric cancer glioblastoma
  • hematological malignancies renal carcinoma or ovarian cancer.
  • the cancer is a chemotherapy resistant cancer.
  • the disclosure also provides methods for treating a cancer in a subject, methods of reducing the rate of the increase of volume of a tumor in a subject over time, methods of reducing the risk of developing a metastasis, or methods of reducing the risk of developing an additional metastasis in a subject.
  • the treatment can halt, slow, retard, or inhibit progression of a cancer.
  • the treatment can result in the reduction of in the number, severity, and/or duration of one or more symptoms of the cancer in a subject.
  • the disclosure features methods that include administering a therapeutically effective amount of antibodies, the antigen-binding fragments thereof, or an antibody drug conjugates disclosed herein to a subject in need thereof, e.g., a subject having, or identified or diagnosed as having, a cancer, e.g., solid tumor, lung cancer (e.g., non-small cell lung cancer, lung adenocarcinoma, or lung carcinoma) , gastric cancer (e.g., gastric carcinoma) , skin cancer (e.g., skin carcinoma) , colorectal cancer, breast cancer, head and neck cancer, ovarian cancer, prostate cancer, thyroid cancer, pancreatic cancer, CNS cancer, liver cancer, nasopharynx cancer, or brain cancer.
  • a cancer e.g., solid tumor
  • lung cancer e.g., non-small cell lung cancer, lung adenocarcinoma, or lung carcinoma
  • gastric cancer e.g., gastric carcinoma
  • skin cancer e.g., skin carcinoma
  • the terms “subject” and “patient” are used interchangeably throughout the specification and describe an animal, human or non-human, to whom treatment according to the methods of the present invention is provided.
  • Veterinary and non-veterinary applications are contemplated by the present invention.
  • Human patients can be adult humans or juvenile humans (e.g., humans below the age of 18 years old) .
  • patients include but are not limited to mice, rats, hamsters, guinea-pigs, rabbits, ferrets, cats, dogs, and primates.
  • non-human primates e.g., monkey, chimpanzee, gorilla, and the like
  • rodents e.g., rats, mice, gerbils, hamsters, ferrets, rabbits
  • lagomorphs e.g., swine (e.g., pig, miniature pig)
  • equine canine, feline, bovine, and other domestic, farm, and zoo animals.
  • compositions and methods disclosed herein can be used for treatment of patients at risk for a cancer.
  • Patients with cancer can be identified with various methods known in the art.
  • an “effective amount” is meant an amount or dosage sufficient to effect beneficial or desired results including halting, slowing, retarding, or inhibiting progression of a disease, e.g., a cancer.
  • An effective amount will vary depending upon, e.g., an age and a body weight of a subject to which the antibody, antigen binding fragment, antibody-drug conjugates, antibody-encoding polynucleotide, vector comprising the polynucleotide, and/or compositions thereof is to be administered, a severity of symptoms and a route of administration, and thus administration can be determined on an individual basis.
  • an effective amount can be administered in one or more administrations.
  • an effective amount of an antibody, an antigen binding fragment, or an antibody-drug conjugate is an amount sufficient to ameliorate, stop, stabilize, reverse, inhibit, slow and/or delay progression of an autoimmune disease or a cancer in a patient or is an amount sufficient to ameliorate, stop, stabilize, reverse, slow and/or delay proliferation of a cell (e.g., a biopsied cell, any of the cancer cells described herein, or cell line (e.g., a cancer cell line) ) in vitro.
  • a cell e.g., a biopsied cell, any of the cancer cells described herein, or cell line (e.g., a cancer cell line)
  • an effective amount of an antibody, antigen binding fragment, or antibody-drug conjugate may vary, depending on, inter alia, patient history as well as other factors such as the type (and/or dosage) of antibody used.
  • Effective amounts and schedules for administering the antibodies, antibody-encoding polynucleotides, antibody-drug conjugates, and/or compositions disclosed herein may be determined empirically, and making such determinations is within the skill in the art.
  • the dosage that must be administered will vary depending on, for example, the mammal that will receive the antibodies, antibody-encoding polynucleotides, antibody-drug conjugates, and/or compositions disclosed herein, the route of administration, the particular type of antibodies, antibody-encoding polynucleotides, antigen binding fragments, antibody-drug conjugates, and/or compositions disclosed herein used and other drugs being administered to the mammal.
  • a typical daily dosage of an effective amount of an antibody, the antigen-binding fragment thereof, or antibody-drug conjugate is 0.01 mg/kg to 100 mg/kg. In some embodiments, the dosage can be less than 100 mg/kg, 30 mg/kg, 20 mg/kg, 10 mg/kg, 9 mg/kg, 8 mg/kg, 7 mg/kg, 6 mg/kg, 5 mg/kg, 4 mg/kg, 3 mg/kg, 2 mg/kg, 1 mg/kg, 0.5 mg/kg, or 0.1 mg/kg.
  • the dosage can be greater than 10 mg/kg, 9 mg/kg, 8 mg/kg, 7 mg/kg, 6 mg/kg, 5 mg/kg, 4 mg/kg, 3 mg/kg, 2 mg/kg, 1 mg/kg, 0.5 mg/kg, 0.1 mg/kg, 0.05 mg/kg, or 0.01 mg/kg.
  • the dosage is about or at least 10 mg/kg, 9 mg/kg, 8 mg/kg, 7 mg/kg, 6 mg/kg, 5 mg/kg, 4 mg/kg, 3 mg/kg, 2 mg/kg, 1 mg/kg, 0.9 mg/kg, 0.8 mg/kg, 0.7 mg/kg, 0.6 mg/kg, 0.5 mg/kg, 0.4 mg/kg, 0.3 mg/kg, 0.2 mg/kg, or 0.1 mg/kg.
  • the at least one antibody, the antigen-binding fragment thereof, antibody-drug conjugates, or pharmaceutical composition e.g., any of the antibodies, antigen-binding fragments, antibody-drug conjugates described herein
  • at least one additional therapeutic agent can be administered to the subject at least once a week (e.g., once a week, twice a week, three times a week, four times a week, once a day, twice a day, or three times a day) .
  • at least two different antibodies and/or antigen-binding fragments are administered in the same composition (e.g., a liquid composition) .
  • At least one antibody, the antigen-binding fragment thereof, or antibody-drug conjugate, and at least one additional therapeutic agent are administered in the same composition (e.g., a liquid composition) .
  • the at least one antibody or antigen-binding fragment and the at least one additional therapeutic agent are administered in two different compositions (e.g., a liquid composition containing at least one antibody or antigen-binding fragment and a solid oral composition containing at least one additional therapeutic agent) .
  • the at least one additional therapeutic agent is administered as a pill, tablet, or capsule.
  • the at least one additional therapeutic agent is administered in a sustained-release oral formulation.
  • the one or more additional therapeutic agents can be administered to the subject prior to, or after administering the at least one antibody, antigen-binding antibody fragment, antibody-drug conjugate, or pharmaceutical composition (e.g., any of the antibodies, antigen-binding antibody fragments, or pharmaceutical compositions described herein) .
  • the one or more additional therapeutic agents and the at least one antibody, antigen-binding antibody fragment, antibody-drug conjugate, or pharmaceutical composition are administered to the subject such that there is an overlap in the bioactive period of the one or more additional therapeutic agents and the at least one antibody or antigen-binding fragment (e.g., any of the antibodies or antigen-binding fragments described herein) in the subject.
  • the subject can be administered the at least one antibody, antigen-binding antibody fragment, antibody-drug conjugate, or pharmaceutical composition (e.g., any of the antibodies, antigen-binding antibody fragments, or pharmaceutical compositions described herein) over an extended period of time (e.g., over a period of at least 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 1 year, 2 years, 3 years, 4 years, or 5 years) .
  • a skilled medical professional may determine the length of the treatment period using any of the methods described herein for diagnosing or following the effectiveness of treatment (e.g., the observation of at least one symptom of cancer) .
  • a skilled medical professional can also change the identity and number (e.g., increase or decrease) of antibodies or antigen-binding antibody fragments, antibody-drug conjugates (and/or one or more additional therapeutic agents) administered to the subject and can also adjust (e.g., increase or decrease) the dosage or frequency of administration of at least one antibody or antigen-binding antibody fragment (and/or one or more additional therapeutic agents) to the subject based on an assessment of the effectiveness of the treatment (e.g., using any of the methods described herein and known in the art) .
  • one or more additional therapeutic agents can be administered to the subject.
  • the additional therapeutic agent can comprise one or more inhibitors selected from the group consisting of an inhibitor of B-Raf, an PD-1 inhibitor, a CD155 inhibitor, a CD112 inhibitor, a 4-1BB inhibitor, a CD40 inhibitor, an inhibitor of a MEK, an inhibitor of ERK, an inhibitor of K-Ras, an inhibitor of c-Met, an inhibitor of anaplastic lymphoma kinase (ALK) , an inhibitor of a phosphatidylinositol3-kinase (PI3K) , an inhibitor of an Akt, an inhibitor of mTOR, a dual PI3K/mTOR inhibitor, an inhibitor of Bruton's tyrosine kinase (BTK) , and an inhibitor of Isocitrate dehydrogenase 1 (IDH1) and/or Isocitrate dehydrogenase 2 (IDH2) .
  • the additional therapeutic agent can comprise
  • the additional therapeutic agent can comprise one or more inhibitors selected from the group consisting of an TIGIT inhibitor, a CD155 inhibitor, a CD112 inhibitor, a 4-1BB inhibitor, a CD40 inhibitor, an inhibitor of LSD1, an inhibitor of MDM2, an inhibitor of BCL2, an inhibitor of CHK1, an inhibitor of activated hedgehog signaling pathway, and an agent that selectively degrades the estrogen receptor.
  • an TIGIT inhibitor a CD155 inhibitor, a CD112 inhibitor, a 4-1BB inhibitor, a CD40 inhibitor, an inhibitor of LSD1, an inhibitor of MDM2, an inhibitor of BCL2, an inhibitor of CHK1, an inhibitor of activated hedgehog signaling pathway, and an agent that selectively degrades the estrogen receptor.
  • the additional therapeutic agent can comprise one or more therapeutic agents selected from the group consisting of Trabectedin, nab-paclitaxel, Trebananib, Pazopanib, Cediranib, Palbociclib, everolimus, fluoropyrimidine, IFL, regorafenib, Reolysin, Alimta, Zykadia, Sutent, temsirolimus, axitinib, everolimus, sorafenib, Votrient, Pazopanib, IMA-901, AGS-003, cabozantinib, Vinflunine, an Hsp90 inhibitor, Ad-GM-CSF, Temazolomide, IL-2, IFNa, vinblastine, Thalomid, dacarbazine, cyclophosphamide, lenalidomide, azacytidine, bortezomid, amrubicine, carfilzomib, pralatrexate, and
  • the additional therapeutic agent can comprise one or more therapeutic agents selected from the group consisting of an adjuvant, a TLR agonist, tumor necrosis factor (TNF) alpha, IL-1, HMGB1, an IL-10 antagonist, an IL-4 antagonist, an IL-13 antagonist, an IL-17 antagonist, an HVEM antagonist, an ICOS agonist, a treatment targeting CX3CL1, a treatment targeting CXCL9, a treatment targeting CXCL10, a treatment targeting CCL5, an LFA-1 agonist, an ICAM1 agonist, and a Selectin agonist.
  • TNF tumor necrosis factor
  • carboplatin, nab-paclitaxel, paclitaxel, cisplatin, pemetrexed, gemcitabine, FOLFOX, or FOLFIRI are administered to the subject.
  • the additional therapeutic agent is an anti-PD-1 antibody, an anti-PD-L1 antibody, anti-PD-L2 antibody, an anti-LAG-3 antibody, an anti-BTLA antibody, an anti-CTLA4 antibody, an anti-CD40 antibody, an anti-OX40 antibody, an anti-4-1BB antibody, or an anti-GITR antibody.
  • compositions that contain at least one (e.g., one, two, three, or four) of the antibodies or antigen-binding fragments described herein. Two or more (e.g., two, three, or four) of any of the antibodies or antigen-binding fragments described herein can be present in a pharmaceutical composition in any combination.
  • the pharmaceutical compositions may be formulated in any manner known in the art.
  • compositions are formulated to be compatible with their intended route of administration (e.g., intravenous, intraarterial, intramuscular, intradermal, subcutaneous, or intraperitoneal) .
  • the compositions can include a sterile diluent (e.g., sterile water or saline) , a fixed oil, polyethylene glycol, glycerine, propylene glycol or other synthetic solvents, antibacterial or antifungal agents, such as benzyl alcohol or methyl parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like, antioxidants, such as ascorbic acid or sodium bisulfite, chelating agents, such as ethylenediaminetetraacetic acid, buffers, such as acetates, citrates, or phosphates, and isotonic agents, such as sugars (e.g., dextrose) , polyalcohols (e.g., mannitol or
  • Liposomal suspensions can also be used as pharmaceutically acceptable carriers (see, e.g., U.S. Patent No. 4,522,811) .
  • Preparations of the compositions can be formulated and enclosed in ampules, disposable syringes, or multiple dose vials. Where required (as in, for example, injectable formulations) , proper fluidity can be maintained by, for example, the use of a coating, such as lecithin, or a surfactant.
  • Absorption of the antibody or antigen-binding fragment thereof can be prolonged by including an agent that delays absorption (e.g., aluminum monostearate and gelatin) .
  • controlled release can be achieved by implants and microencapsulated delivery systems, which can include biodegradable, biocompatible polymers (e.g., ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid; Alza Corporation and Nova Pharmaceutical, Inc. ) .
  • biodegradable, biocompatible polymers e.g., ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid; Alza Corporation and Nova Pharmaceutical, Inc.
  • compositions containing one or more of any of the antibodies or antigen-binding fragments described herein can be formulated for parenteral (e.g., intravenous, intraarterial, intramuscular, intradermal, subcutaneous, or intraperitoneal) administration in dosage unit form (i.e., physically discrete units containing a predetermined quantity of active compound for ease of administration and uniformity of dosage) .
  • parenteral e.g., intravenous, intraarterial, intramuscular, intradermal, subcutaneous, or intraperitoneal
  • dosage unit form i.e., physically discrete units containing a predetermined quantity of active compound for ease of administration and uniformity of dosage
  • Toxicity and therapeutic efficacy of compositions can be determined by standard pharmaceutical procedures in cell cultures or experimental animals (e.g., monkeys) .
  • Agents that exhibit high therapeutic indices are preferred. Where an agent exhibits an undesirable side effect, care should be taken to minimize potential damage (i.e., reduce unwanted side effects) .
  • Toxicity and therapeutic efficacy can be determined by other standard pharmaceutical procedures.
  • a therapeutically effective amount of the one or more (e.g., one, two, three, or four) antibodies or antigen-binding fragments thereof (e.g., any of the antibodies or antibody fragments described herein) will be an amount that treats the disease (e.g., kills cancer cells) in a subject (e.g., a human subject identified as having cancer) , or a subject identified as being at risk of developing the disease (e.g., a subject who has previously developed cancer but now has been cured) , decreases the severity, frequency, and/or duration of one or more symptoms of a disease in a subject (e.g., a human) .
  • any of the antibodies or antigen-binding fragments described herein can be determined by a health care professional or veterinary professional using methods known in the art, as well as by the observation of one or more symptoms of disease in a subject (e.g., a human) . Certain factors may influence the dosage and timing required to effectively treat a subject (e.g., the severity of the disease or disorder, previous treatments, the general health and/or age of the subject, and the presence of other diseases) .
  • Exemplary doses include milligram or microgram amounts of any of the antibodies or antigen-binding fragments, or antibody-drug conjugates described herein per kilogram of the subject’s weight (e.g., about 1 ⁇ g/kg to about 500 mg/kg; about 100 ⁇ g/kg to about 500 mg/kg; about 100 ⁇ g/kg to about 50 mg/kg; about 10 ⁇ g/kg to about 5 mg/kg; about 10 ⁇ g/kg to about 0.5 mg/kg; or about 0.1 mg/kg to about 0.5 mg/kg) . While these doses cover a broad range, one of ordinary skill in the art will understand that therapeutic agents, including antibodies and antigen-binding fragments thereof, vary in their potency, and effective amounts can be determined by methods known in the art.
  • relatively low doses are administered at first, and the attending health care professional or veterinary professional (in the case of therapeutic application) or a researcher (when still working at the development stage) can subsequently and gradually increase the dose until an appropriate response is obtained.
  • the specific dose level for any particular subject will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, gender, and diet of the subject, the time of administration, the route of administration, the rate of excretion, and the half-life of the antibody or antibody fragment in vivo.
  • compositions can be included in a container, pack, or dispenser together with instructions for administration.
  • the disclosure also provides methods of manufacturing the antibodies or antigen binding fragments thereof, or antibody-drug conjugates for various uses as described herein.
  • Human TIGIT protein (hTIGIT-Fc, ACRO Biosystems Inc., Cat#: TIT-H5254) and Cynomolgus/Rhesus macaque TIGIT protein (cynTIGIT-Fc, ACRO Biosystems Inc., Cat#: TIT-C5254) , or expression plasmids encoding human TIGIT or Rhesus monkey TIGIT were emulsified with adjuvants, and was used to immunize RenMab TM .
  • RenMab TM mice have both a humanized heavy chain immunoglobulin locus and a humanized kappa chain immunoglobulin locus.
  • the heavy chain immunoglobulin locus is a region on the chromosome that contains genes for the heavy chains of antibodies.
  • the locus includes IGHV (variable) , IGHD (diversity) , IGHJ (joining) , and heavy chain constant domain genes.
  • the kappa chain immunoglobulin locus is a region on the chromosome that contains genes that encode the light chains of antibodies (kappa chain) .
  • the kappa chain immunoglobulin locus includes IGKV (variable) , IGKJ (joining) , and light chain constant domain genes.
  • IGKV variable
  • IGKJ joining
  • TIGIT protein was injected by intraperitoneal injection, and the CHO-S cells expressing human TIGIT antigen was injected through the tail vein.
  • Antigen-specific immune cells were isolated from the immunized mice to further obtain anti-TIGIT antibodies or to obtain the light chain and heavy chain variable region sequences of the anti-TIGIT antibodies.
  • single cell technology e.g., using Optofluidic System, Berkeley Lights Inc.
  • reverse transcription and PCR sequencing were used to obtain antibody variable region sequences.
  • the obtained variable region sequences were cloned into a vector containing a sequence encoding the human IgG1 constant region for antibody expression. Binding of the expressed antibodies to TIGIT was verified by FACS.
  • Exemplary antibodies obtained by this method included: 5F1, 15C5, 36F3 and 36H7.
  • phage display was performed to screen and find monoclonal antibodies that was antigen-specific.
  • 9H11 was the exemplary antibody obtained by this method.
  • VH and VL CDR 1-3 sequences of 5F1, 9H11, 15C5, 36F3 and 36H7 are shown in FIG. 1 and FIG. 2.
  • the VH and VL regions of 5F1, 9H11, 15C5, 36F3 and 36H7 are shown in FIG. 3.
  • a reference antibody targeting TIGIT labeled as PC was included in the following experiments, for comparative purposes.
  • the antibody PC has VH/VL sequences as shown in SEQ ID NOs: 57-58, and human IgG1 constant region.
  • the constant region of above antibodies can also include some mutations.
  • LALA mutations L234A and L235A mutations in EU numbering
  • LALA is added to the name.
  • the antibody is named as 5F1-LALA.
  • the “EN” in VH CDR of the antibodies described herein was mutated to “ED” to create CDR-modified VHs.
  • the “EN” in VH CDR2 of 15C5 was mutated to “ED” , and the resulting CDR-modified VH is named 15C5-ND VH.
  • the sequence of the CDR-modified VH can be found in FIG. 3.
  • VH CDR of the antibodies described herein was mutated to “YG” to create CDR-modified VHs.
  • the “YS” in VH CDR2 of 15C5 was mutated to “YG”
  • the resulting CDR-modified VH is named 15C5-SG VH.
  • the sequence of the CDR-modified VH can be found in FIG. 3.
  • binding affinities of the anti-TIGIT antibodies to His-tagged TIGIT protein of human hTIGIT-His, ACRO biosystems, Cat#: TIT-H52H3 or monkey (cynTIGIT-His, ACRO biosystems, Cat#: TIT-C5223) were verified using surface plasmon resonance (SPR) using a Biacore TM (Biacore, Inc., Piscataway N.J. ) 8K biosensor equipped with pre-immobilized Protein A sensor chips.
  • SPR surface plasmon resonance
  • Purified anti-TIGIT antibodies was captured on the Protein A chip (Series S Sensor Chip Protein A) for the detection. 1 ⁇ g/mL or 2 ⁇ g/mL purified anti-TIGIT antibodies was loaded at 10 ⁇ L/min to bind to the recombinant hTIGIT-His or cynTIGIT-His with the concentration of 100 nM. The flow rate was 30 ⁇ L/min, the binding and dissociation time were set to 180s and 400s, respectively. The chip was regenerated after the last injection of each titration with a glycine solution (pH2.0) at 30 ⁇ L/min for 30 seconds.
  • a glycine solution pH2.0
  • the experiment was performed to test whether the anti-TIGIT antibodies can block the TIGIT/CD155 pathway.
  • CHO-S-hTIGIT cells obtained by transfecting CHO-S cells with a vector expressing human TIGIT (SEQ ID NO: 56) ) were seeded in a 96-well plate (cell density 2 ⁇ 10 5 cells/well) .
  • the anti-TIGIT antibodies 5F1, 9H11, 15C5, 36F3, 36H7 and PC were serially diluted (2-fold) with the highest concentration at 50 ⁇ g/mL, respectively.
  • the assay buffer was phosphate buffer saline (PBS) . 30 ⁇ L antibody was added to each well. PBS was added to each well as a negative control.
  • the above 96-well plate was incubated in a 4°C incubator for 15 minutes.
  • the ligand biotinylated human CD155 protein (ACRO biosystems, Cat#: TIT-C5223) was added to the plate with a concentration of 0.3 ⁇ g/mL, and incubated in a 4°C incubator for 20 minutes. Then, after one wash with PBS, the cells were incubated with the secondary antibody Anti-hIgG-Fc-Alexa Fluor 647 (Jackson, Cat#: 109-606-170) at 1: 5000 dilution and PE Stretavidin at 1: 100 dilution at 4°C for 15 minutes before flow cytometry analysis. The mean fluorescence intensity (MFI) was determined. A fitting curve was obtained using antibody concentration (log10, ⁇ g/mL) as the X-axis and MFI as the Y-axis. The results are shown in FIGs. 4A-4C.
  • HBS-EP+buffer 10 mM4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid (HEPES) , 150 mM NaCl, 3 mM ethylenediaminetetraacetic acid (EDTA) and 0.05%P20, pH7.4) diluted from HBS-EP+buffer (10 ⁇ ) was used as the running buffer throughout the experiment.
  • the ligand hTIGIT-His protein with a concentration of 5 ⁇ g/mL was captured at a flow rate of 10 ⁇ L/min, and 200 nM of antibody was injected at a flow rate of 30 ⁇ L/min to bind the ligand.
  • Another antibody was injected under the same conditions to determine whether the binding of different antibodies interfered with each other. The binding time was 300 s for each antibody.
  • the binding value of each antibody was obtained using Data Analysis HT 12.0. To quantify the interference of one antibody binding to another, a binding ratio was calculated to compare each pair of antibodies. The binding ratio is defined as the binding value of the second antibody, divided by the binding value of the first antibody. The threshold to determine whether the two antibodies can interfere with each other is 0.5. For example, a binding ratio over 0.5 indicates that the second antibody can still bind to TIGIT subsequent to the binding of the first antibody. As such, the two antibodies may recognize different epitopes.
  • Anti-TIGIT antibodies were diluted to 2 mg/mL using a buffer at pH6.0 (3 mg/mL histidine, 80 mg/mL sucrose, and 0.2 mg/mL Tween 80) .
  • the diluted antibodies were kept in sealed Eppendorftubes at 40 ⁇ 2°C (hereinafter referred to as 40°C) for 7 days, and their thermal stability was evaluated.
  • the antibody samples were diluted to 1 mg/mL with purified water and an Agilent 1290 chromatograph system (connected with XBridge Protein BEH SEC column ( Waters Corporation) ) was used.
  • the following parameters were used: mobile phase: 0.1Mphosphate buffer (PB) +10%ACN, pH7.4; flow rate: 1.8 ml/min; column temperature: 25°C; detection wavelength: 280 nm, 220nm; injection volume: 10 ⁇ L; sample tray temperature: about 4°C; and running time: 7 minutes.
  • the Maurice cIEF Method Development Kit (Protein Simple, Cat#: PS-MDK01-C) was used for sample preparation. Specifically, 40 ⁇ g protein sample was mixed with the following reagents in the kit: 1 ⁇ L Maurice cIEF pI Marker-4.05, 1 ⁇ L Maurice cIEF pI Marker-9.99, 35 ⁇ L 1%Methyl Cellulose Solution, 2 ⁇ L Maurice cIEF 500 mM Arginine, 4 ⁇ L Ampholytes (Pharmalyte pH ranges 3-10) , and water (added to make a final volume of 100 ⁇ L) .
  • Maurice cIEF Cartridges PS-MC02-C were used to generate imaging capillary isoelectric focusing spectra. The sample was focused for a total of 10 minutes. The analysis software installed on the instrument was used to analyze the absorbance of the 280 nm-focused protein.
  • mice The pharmacokinetic properties of the anti-TIGIT antibodies were determined in C57BL/6 mice. Specifically, the mice were placed into four groups (3 mice per group) , and administered with 3 mg/kg of 9H11, 15C5-SG, 36F3 or PC by intravenous injection. Blood samples were collected 4 days before administration and 15 minutes, 4 hr, 1 day, 2 days, 4 days and 6 days after administration.
  • the serum levels of human antibodies were determined by sandwich ELISA. Briefly, Goat Anti-Human IgG (H+L) (Jackson ImmunoResearch Inc., Cat#: 109-005-088) was diluted to a final concentration of 2000 ng/mL, added to a 96-well plate (ELISA plate) at 100 ⁇ L/well, and then incubated overnight at 4°C. After the incubation, the plate was washed with PBS-T buffer (PBS supplemented with Tween TM 20) 4 times. Antibody-unbound areas were blocked with 2%BSA (bovine serum albumin) for 1 hours at 37°C. Afterwards, the plate was washed with PBS-T buffer 4 times.
  • Goat Anti-Human IgG H+L
  • ELISA plate 96-well plate
  • TMB tetramethylbenzidine
  • the experiment was performed to test ADCC (antibody-dependent cellular cytotoxicity) activity of anti-TIGIT antibodies. Specifically, effector cells Jurkat-luc-hCD16A (obtained by transfecting Jurkat cells with a vector expressing human CD16A (SEQ ID NO: 60) ) and target cells CHO-S-TIGIT (obtained by transfecting CHO-S cells with a vector expressing human TIGIT (SEQ ID NO: 56) ) at a E: T ratio of 4: 1 were co-incubated with serially diluted anti-TIGIT antibodies for 6hours. After the incubation, 75 ⁇ L luciferase substrate was added and incubated at room temperature for 5 minutes. Then, the mixture was placed into a 96 microplate luminometer to test the cell killing activity. EC50 was caculated.
  • a humanized TIGIT mouse model (hTIGIT mice) was engineered to express a chimeric TIGIT protein (SEQ ID NO: 59) wherein a part of the extracellular region of the mouse TIGIT protein was replaced with the corresponding human TIGIT extracellular region.
  • SEQ ID NO: 59 a chimeric TIGIT protein
  • mice About 5 ⁇ 10 5 MC38 cells were injected subcutaneously in hTIGIT mice, and when the tumor volume reached about 100-150 mm 3 , the mice were divided to a control group and four treatment groups based on tumor size (6 mice per group) .
  • the treatment groups were randomly selected for 9H11, 36F3, 36H7 or PC treatment.
  • the control group mice were injected with phosphate buffer saline (PBS) .
  • the tumor volumes were measured twice a week and the body weights of the mice were weighed as well. Euthanasia was performed when the tumor volume of a mouse reached 3000 mm 3 . Details of the administration scheme are shown in the table below.
  • TGI tumor growth inhibition percentage
  • mice in different groups all increased. On the day of group assignment (Day 0) , the average weight of each group was in the range of 21.8 g-22.1 g; At the end of the experiment (Day 21) , the average weight of each group was in the range of 23.9 g-24.8 g. Thus, the average weight change of each group was in the range of 109.4%-112.4%. The results showed that the tested antibodies were well tolerated and were not obviously toxic to the mice.
  • the tumor sizes in groups treated with the antibodies are shown in FIG. 7.
  • the table below summarizes the results for this experiment, including the tumor volumes on the day of grouping (day 0) , 11 days after grouping (day 11) and at the end of the experiment (day 21) ; the survival rate of the mice; Tumor Growth Inhibition value (TGI%) ; and the statistical differences (P value) of tumor volume and body weight between the treatment and control groups.
  • the tumor growths in the treatment groups (G2 to G5) were inhibited to different extents.
  • 36F3, 36H7 and 9H11 exhibited better tumor inhibitory effect as compared to the positive control PC.
  • mice in different groups all increased. On the day of group assignment (Day 0) , the average weight of each group was in the range of 21.1 g-21.5 g; on Day 21, the average weight of each group was in the range of 23.1 g-24.5 g. Thus, the average weight change of each group was in the range of 107.9%-112.9%. The results showed that the tested antibodies were well tolerated and were not obviously toxic to the mice.
  • the tumor sizes in groups treated with the antibodies are shown in FIG. 8.
  • the table below summarizes the results for this experiment, including the tumor volumes on Day 0, Day 11 and Day 21; the survival rate of the mice, TGI%, and P value of tumor volume and body weight between the treatment and control groups on Day 21.
  • the tumor growths in the treatment groups (G2-G6) were inhibited to different extents.
  • 9H11 and 36F3 inhibited tumor growth with higher TGI%at both the 3mg/kg dose level and the 10mg/kg dose level than that of the positive control PC at 10 mg/kg dosage.
  • mice About 5 ⁇ 10 5 MC38 cells were injected subcutaneously in hTIGIT mice, and when the tumor volume grew to about 100-150 mm 3 , the mice were assigned to a control group and two treatment groups based on tumor size (5 mice per group) . The treatment groups were randomly selected for 15C5 or PC treatment. The control group mice were injected with PBS. The tumor volumes were measured twice a week and the body weights of the mice were weighed as well. Euthanasia was performed when the tumor volume of a mouse reached 3000 mm 3 . Details of the administration scheme are shown in the table below.
  • mice in different groups all increased. On the day of group assignment (Day 0) , the average weight of each group was 19.9 g. On Day 21, the average weight of each group was in the range of 22.7 g-23.8 g. The average weight change of each group was in the range of 114.0%-120.0%. The results showed that the tested antibodies were well tolerated and were not obviously toxic to the mice.
  • the tumor sizes in groups treated with the antibodies are shown in FIG. 9.
  • the table below summarizes the results for this experiment, including the tumor volumes on Day 0, Day 11 and Day 21; the survival rate of the mice, TGI%and P value of tumor volume and body weight between the treatment and control groups.
  • 15C5 exhibited better tumor inhibitory effect as compared to the positive control PC.

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Abstract

L'invention concerne des anticorps anti-TIGIT ou des fragments de liaison à l'antigène de ceux-ci, et les conjugués anticorps-médicament associés.
PCT/CN2024/119890 2023-09-21 2024-09-20 Anticorps anti-tigit et leurs utilisations Pending WO2025061119A1 (fr)

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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019062832A1 (fr) * 2017-09-29 2019-04-04 江苏恒瑞医药股份有限公司 Anticorps tigit, fragment de liaison à l'antigène de celui-ci, et son utilisation médicale
US20190328907A1 (en) * 2016-08-31 2019-10-31 Beijing Biocytogen Co., Ltd. Genetically Modified Non-Human Animal With Human Or Chimeric TIGIT
US20220363754A1 (en) * 2019-07-15 2022-11-17 Shanghai Junshi Biosciences Co., Ltd. Anti-tigit antibodies and application thereof
US20220389095A1 (en) * 2021-05-04 2022-12-08 Agenus Inc. Anti-tigit antibodies and methods of use thereof
US20220396617A1 (en) * 2021-06-10 2022-12-15 Beijing Mabworks Biotech Co. Ltd. Antibodies binding tigit and uses thereof
US20230272064A1 (en) * 2020-04-29 2023-08-31 Acroimmune Biotech Co., Ltd. Monoclonal antibody binding to tigit antigen, preparation method and use thereof

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20190328907A1 (en) * 2016-08-31 2019-10-31 Beijing Biocytogen Co., Ltd. Genetically Modified Non-Human Animal With Human Or Chimeric TIGIT
WO2019062832A1 (fr) * 2017-09-29 2019-04-04 江苏恒瑞医药股份有限公司 Anticorps tigit, fragment de liaison à l'antigène de celui-ci, et son utilisation médicale
US20220363754A1 (en) * 2019-07-15 2022-11-17 Shanghai Junshi Biosciences Co., Ltd. Anti-tigit antibodies and application thereof
US20230272064A1 (en) * 2020-04-29 2023-08-31 Acroimmune Biotech Co., Ltd. Monoclonal antibody binding to tigit antigen, preparation method and use thereof
US20220389095A1 (en) * 2021-05-04 2022-12-08 Agenus Inc. Anti-tigit antibodies and methods of use thereof
US20220396617A1 (en) * 2021-06-10 2022-12-15 Beijing Mabworks Biotech Co. Ltd. Antibodies binding tigit and uses thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
SOLOMON, B.L. ET AL.: "TIGIT: a novel immunotherapy target moving from bench to bedside", CANCER IMMUNOLOGY, IMMUNOTHERAPY, vol. 67, 19 September 2018 (2018-09-19), XP036618739, DOI: 10.1007/s00262-018-2246-5 *

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