WO2023143227A1 - 抗cd73抗体及应用 - Google Patents

抗cd73抗体及应用 Download PDF

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Publication number
WO2023143227A1
WO2023143227A1 PCT/CN2023/072527 CN2023072527W WO2023143227A1 WO 2023143227 A1 WO2023143227 A1 WO 2023143227A1 CN 2023072527 W CN2023072527 W CN 2023072527W WO 2023143227 A1 WO2023143227 A1 WO 2023143227A1
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Prior art keywords
seq
sequence
sequence shown
identity
antibody
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PCT/CN2023/072527
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English (en)
French (fr)
Inventor
丁焕弟
惠希武
姚兵
淡墨
靳子怡
郭青娟
袁灿
李文宾
王超
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石药集团巨石生物制药有限公司
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Publication of WO2023143227A1 publication Critical patent/WO2023143227A1/zh

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    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • 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

Definitions

  • the present application generally relates to the field of biopharmaceuticals, in particular, the present application relates to novel anti-CD73 antibodies or antigen-binding fragments thereof, pharmaceutical compositions comprising said antibodies or antigen-binding fragments thereof, and pharmaceuticals for said antibodies or antigen-binding fragments thereof learning purposes.
  • CD73 is an extracellular-5'-nucleotidase encoded by the NT5E gene, which includes two parts, the N-terminal and the C-terminal, and is anchored to the outer surface of the cell membrane by glycosylphosphatidylinositol (GPI) to form a homodimer.
  • GPI glycosylphosphatidylinositol
  • CD73 is widely expressed on the surface of human endothelial cells, lymphocytes and other cells. It is highly expressed in various tumor tissues such as gastric cancer, renal cancer, prostate cancer, breast cancer (such as triple-negative breast cancer), non-small cell adenocarcinoma, and indicates poor prognosis.
  • CD73 can hydrolyze extracellular adenosine monophosphate (AMP) to adenosine.
  • AMP extracellular adenosine monophosphate
  • Adenosine is a powerful immunosuppressive molecule that inhibits the function of protective immune cells (such as effector T cells, NK cells, DCs, and B cells) while maintaining regulatory immune cells (such as Treg, MDSC, TAM and CAF) to weaken anti-tumor immunity and participate in tumor immune escape.
  • protective immune cells such as effector T cells, NK cells, DCs, and B cells
  • regulatory immune cells such as Treg, MDSC, TAM and CAF
  • CD73 can also directly stimulate tumor cell proliferation, migration, invasion and tumor angiogenesis, and inhibition of extracellular nucleotidase CD73 can reverse adenosine-mediated immunosuppression. Studies have found that hypoxia, radiotherapy and chemotherapy lead to the upregulation of PDL1, CD73 and CD47 expression.
  • CD73 has the potential to be applied clinically as monotherapy or combination therapy.
  • Medimmune s oleclumab
  • I-Mab uliledlimab.
  • the fastest-growing CD73 antibody is in the third phase of clinical trials, and there is no marketed product. Therefore, the development and application of CD73 antibodies are urgently needed in this field.
  • the application provides an anti-CD73 antibody or an antigen-binding portion thereof, the anti-CD73 antibody or an antigen-binding portion thereof comprising a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising a heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3), the light chain variable region includes light chain CDR1 (LCDR1), light chain CDR2 (LCDR2) and light chain CDR3 (LCDR3), where:
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1
  • the sequence of HCDR2 is SEQ ID NO.2
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or with the sequence shown in SEQ ID NO.
  • sequence shown in 3 has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR1 is the sequence shown in SEQ ID NO.5 or has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR2 is the sequence shown in SEQ ID NO.6 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.6 99% identity
  • sequence of LCDR3 is the sequence shown in SEQ ID NO.7 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.7; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1
  • the sequence of HCDR2 is SEQ ID NO.17
  • the sequence shown may have at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.17
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or with the sequence shown in SEQ ID NO.
  • the sequence shown in 3 has at least 80%, 85%, 90%, 95% or 99% identity
  • the sequence of LCDR1 is the sequence shown in SEQ ID NO.5 or has at least 80%, 85%, 90%, 95% or 99% identity
  • the sequence of LCDR2 is the sequence shown in SEQ ID NO.32 or has at least 80%, 85%, 90%, 95% or 99% identity
  • the sequence of LCDR3 is SEQ ID The sequence shown in NO.7 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.7; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1, and the sequence of HCDR2 is SEQ ID NO.17
  • the sequence shown is at least 80%, 85%, 90%, 95% or 99% identical to the sequence shown in SEQ ID NO.17
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or the same as SEQ ID NO.
  • sequence shown in 3 has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR1 is the sequence shown in SEQ ID NO.5 or has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR2 is the sequence shown in SEQ ID NO.6 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.6 99% identity
  • sequence of LCDR3 is the sequence shown in SEQ ID NO.7 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.7; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1
  • the sequence of HCDR2 is SEQ ID NO.2
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or with the sequence shown in SEQ ID NO.
  • sequence shown in 3 has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR1 is the sequence shown in SEQ ID NO.5 or has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR2 is the sequence shown in SEQ ID NO.32 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.32 99% identity
  • sequence of LCDR3 is the sequence shown in SEQ ID NO.7 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.7; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1
  • the sequence of HCDR2 is SEQ ID NO.21
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or with the sequence shown in SEQ ID NO.
  • sequence shown in 3 has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR1 is the sequence shown in SEQ ID NO.5 or has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR2 is the sequence shown in SEQ ID NO.32 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.32 99% identity
  • sequence of LCDR3 is the sequence shown in SEQ ID NO.7 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.7; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1
  • the sequence of HCDR2 is SEQ ID NO.21
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or with the sequence shown in SEQ ID NO.
  • sequence shown in 3 has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR1 is the sequence shown in SEQ ID NO.5 or has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR2 is the sequence shown in SEQ ID NO.6 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.6 99% identity
  • sequence of LCDR3 is the sequence shown in SEQ ID NO.7 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.7; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1
  • the sequence of HCDR2 is SEQ ID NO.23
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or with the sequence shown in SEQ ID NO.
  • sequence shown in 3 has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR1 is the sequence shown in SEQ ID NO.5 or has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR2 is the sequence shown in SEQ ID NO.32 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.32 99% identity
  • sequence of LCDR3 is the sequence shown in SEQ ID NO.7 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.7; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1, and the sequence of HCDR2 is SEQ ID NO.23 The sequence shown or with the SEQ ID The sequence shown in NO.23 has at least 80%, 85%, 90%, 95% or 99% identity, and the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or has at least 80% identity with the sequence shown in SEQ ID NO.3 %, 85%, 90%, 95% or 99% identity, the sequence of LCDR1 is the sequence shown in SEQ ID NO.5 or has at least 80%, 85%, 90%, 95% with the sequence shown in SEQ ID NO.5 % or 99% identity, the sequence of LCDR2 is the sequence shown in SEQ ID NO.6 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.6, the sequence of LCDR3 The sequence is or is
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.9 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.9
  • the sequence of HCDR2 is SEQ ID NO.10
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.11 or with the sequence shown in SEQ ID NO.
  • sequence shown in 11 has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR1 is the sequence shown in SEQ ID NO.13 or has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR2 is the sequence shown in SEQ ID NO.14 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.14 99% identity
  • sequence of LCDR3 is the sequence shown in SEQ ID NO.15 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.15; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.9 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.9
  • the sequence of HCDR2 is SEQ ID NO.10
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.11 or with the sequence shown in SEQ ID NO.
  • the sequence shown in 11 has at least 80%, 85%, 90%, 95% or 99% identity
  • the sequence of LCDR1 is the sequence shown in SEQ ID NO.35 or has at least 80%, 85%, 90%, 95% or 99% identity
  • the sequence of LCDR2 is the sequence shown in SEQ ID NO.14 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.14 99% identity
  • the sequence of LCDR3 is the sequence shown in SEQ ID NO.15 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.15; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.9 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.9
  • the sequence of HCDR2 is SEQ ID NO.27
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.11 or with the sequence shown in SEQ ID NO.
  • the sequence shown in 11 has at least 80%, 85%, 90%, 95% or 99% identity
  • the sequence of LCDR1 is the sequence shown in SEQ ID NO.35 or has at least 80%, 85%, 90%, 95% or 99% identity
  • the sequence of LCDR2 is the sequence shown in SEQ ID NO.14 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.14 99% identity
  • the sequence of LCDR3 is the sequence shown in SEQ ID NO.15 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.15; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.9 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.9
  • the sequence of HCDR2 is SEQ ID NO.27
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.11 or with the sequence shown in SEQ ID NO.
  • sequence shown in 11 has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR1 is the sequence shown in SEQ ID NO.13 or has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR2 is the sequence shown in SEQ ID NO.14 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.14 99% identity
  • sequence of LCDR3 is the sequence shown in SEQ ID NO.15 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.15; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.9 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.9
  • the sequence of HCDR2 is SEQ ID NO.30
  • the sequence shown may have at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.30
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.11 or with the sequence shown in SEQ ID NO.
  • the sequence shown in 11 has at least 80%, 85%, 90%, 95% or 99% identity
  • the sequence of LCDR1 is the sequence shown in SEQ ID NO.35 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.35
  • the sequence of LCDR2 is SEQ ID
  • the sequence of LCDR3 is the sequence shown in SEQ ID NO.15 or with SEQ ID NO.15
  • the sequence shown in ID NO. 15 has at least 80%, 85%, 90%, 95% or 99% identity; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.9 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.9
  • the sequence of HCDR2 is SEQ ID NO.30
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.11 or with the sequence shown in SEQ ID NO.
  • sequence shown in 11 has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR1 is the sequence shown in SEQ ID NO.13 or has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR2 is the sequence shown in SEQ ID NO.14 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.14 99% identity
  • sequence of LCDR3 is the sequence shown in SEQ ID NO.15 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.15;
  • HCDR and LCDR sequences are defined according to Kabat.
  • the present application provides an anti-CD73 antibody or an antigen-binding portion thereof, the anti-CD73 antibody or an antigen-binding portion thereof comprising a heavy chain variable region and a light chain variable region, wherein:
  • the heavy chain variable region has an amino acid sequence shown in SEQ ID NO.4, 18, 19, 20, 22, 24, 12, 25, 26, 28, 29 or 31 or the same as SEQ ID NO.4, 18, 19 , 20, 22, 24, 12, 25, 26, 28, 29 or 31 have an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to the amino acid sequence; and/ or
  • the light chain variable region has an amino acid sequence shown in SEQ ID NO.8, 33, 34, 16, 36, 37 or 38 or the same as shown in SEQ ID NO.8, 33, 34, 16, 36, 37 or 38 Amino acid sequences are amino acid sequences that are at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical.
  • the present application provides an anti-CD73 antibody or an antigen-binding portion thereof, the anti-CD73 antibody or an antigen-binding portion thereof comprising a heavy chain variable region, the heavy chain variable region comprising a heavy chain CDR1 (HCDR1), a heavy chain chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3), wherein:
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1
  • the sequence of HCDR2 is SEQ ID
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or with SEQ ID NO.
  • the sequence shown in ID NO.3 has at least 80%, 85%, 90%, 95% or 99% identity; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1
  • the sequence of HCDR2 is SEQ ID
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or with SEQ ID NO.3
  • the sequence shown in ID NO.3 has at least 80%, 85%, 90%, 95% or 99% identity; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1
  • the sequence of HCDR2 is SEQ ID
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or with SEQ ID NO.
  • the sequence shown in ID NO.3 has at least 80%, 85%, 90%, 95% or 99% identity; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1
  • the sequence of HCDR2 is SEQ ID
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or with SEQ ID NO.3
  • the sequence shown in ID NO.3 has at least 80%, 85%, 90%, 95% or 99% identity one sex; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.9 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.9
  • the sequence of HCDR2 is SEQ ID
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.11 or with the sequence shown in SEQ ID NO.10
  • the sequence shown in ID NO.11 has at least 80%, 85%, 90%, 95% or 99% identity; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.9 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.9
  • the sequence of HCDR2 is SEQ ID
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.11 or with the sequence shown in SEQ ID NO.11
  • the sequence shown in ID NO.11 has at least 80%, 85%, 90%, 95% or 99% identity; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.9 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.9
  • the sequence of HCDR2 is SEQ ID
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.11 or with SEQ ID NO.11
  • the sequence shown in ID NO.11 has at least 80%, 85%, 90%, 95% or 99% identity;
  • HCDR sequence is defined according to Kabat.
  • the present application provides an anti-CD73 antibody or an antigen-binding portion thereof, the anti-CD73 antibody or an antigen-binding portion thereof comprising a light chain variable region, the light chain variable region comprising a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2) and light chain CDR3 (LCDR3), wherein:
  • the sequence of LCDR1 is the sequence shown in SEQ ID NO.5 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.5, and the sequence of LCDR2 is SEQ ID The sequence shown in NO.6 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.6, the sequence of LCDR3 is the sequence shown in SEQ ID NO.7 or with SEQ ID NO. The sequence shown in ID NO.7 has at least 80%, 85%, 90%, 95% or 99% identity; or
  • the sequence of LCDR1 is the sequence shown in SEQ ID NO.5 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.5, and the sequence of LCDR2 is SEQ ID The sequence shown in NO.32 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.32, and the sequence of LCDR3 is the sequence shown in SEQ ID NO.7 or with SEQ ID NO.7 The sequence shown in ID NO.7 has at least 80%, 85%, 90%, 95% or 99% identity; or
  • the sequence of LCDR1 is the sequence shown in SEQ ID NO.13 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.13
  • the sequence of LCDR2 is SEQ ID
  • the sequence of LCDR3 is the sequence shown in SEQ ID NO.15 or with SEQ ID NO.14
  • the sequence shown in ID NO.15 has at least 80%, 85%, 90%, 95% or 99% identity; or
  • the sequence of LCDR1 is the sequence shown in SEQ ID NO.35 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.35
  • the sequence of LCDR2 is SEQ ID
  • the sequence of LCDR3 is the sequence shown in SEQ ID NO.15 or with SEQ ID NO.14
  • the sequence shown in ID NO.15 has at least 80%, 85%, 90%, 95% or 99% identity;
  • LCDR sequence is defined according to Kabat.
  • the present application provides an isolated nucleic acid molecule encoding the anti-CD73 antibody or antigen-binding portion thereof described in the first to fourth aspects.
  • the present application provides a vector containing the nucleic acid molecule described in the fifth aspect.
  • the present application provides a host cell containing the nucleic acid molecule of the fifth aspect or the vector of the sixth aspect.
  • the present application provides an antibody-drug conjugate, which comprises the anti-CD73 antibody or the antigen-binding part thereof described in the first to fourth aspects coupled with a therapeutic agent.
  • the present application provides a pharmaceutical composition, which comprises the anti-CD73 antibody or its antigen-binding portion described in the first aspect to the fourth aspect or the antibody-drug conjugate described in the eighth aspect, and a pharmaceutically acceptable carrier.
  • the present application provides the anti-CD73 antibody or its antigen-binding portion described in the first to fourth aspects, the nucleic acid molecule described in the fifth aspect, the carrier described in the sixth aspect, and the anti-CD73 antibody described in the seventh aspect.
  • the present application provides a method for treating a tumor in an individual, especially a tumor expressing CD73 (CD73+) on the surface of tumor cells, the method comprising administering to the individual a therapeutically effective amount of the first aspect to the fourth aspect.
  • the present application provides a conjugate, which comprises the anti-CD73 antibody or antigen-binding portion thereof described in the first to fourth aspects and a detectable label.
  • the present application provides a fusion protein comprising the anti-CD73 antibody or antigen-binding portion thereof described in the first to fourth aspects.
  • Figure 1 shows the test results of the binding ability of exemplary antibodies of the present application to soluble CD73.
  • Figure 2 shows the test results of the binding ability of exemplary humanized antibodies of the present application to soluble CD73.
  • Figure 3 shows the test results of the binding ability of the exemplary humanized antibody of the present application to CD73 on the surface of human breast cancer MDA-MB-231 cells.
  • Figure 4 shows the test results of the binding ability of the exemplary humanized antibody of the present application to CD73 on the surface of human ovarian cancer SKOV3 cells.
  • Figure 5 shows the test results of the binding ability of the exemplary humanized antibody of the present application to cynomolgus CD73 on the cell surface.
  • Fig. 6 shows the results of the enzyme activity inhibition experiment of the exemplary humanized antibody of the present application on CD73 protein.
  • Fig. 7 shows the results of the enzyme activity inhibition experiment of the exemplary humanized antibody of the present application on the CD73 protein on the cell surface.
  • Figure 8 shows the experimental results of CD73 internalization induced by exemplary humanized antibodies of the present application.
  • Figure 9A shows the detection results of the exemplary humanized antibody of the present application recognizing the N-terminus of CD73 protein.
  • Figure 9B shows the detection results of the exemplary humanized antibody of the present application recognizing the C-terminus of CD73 protein.
  • Figure 10A shows the results of the volume change of xenograft tumors in A375 tumor-bearing mice resulting from the administration of exemplary antibodies of the present application and other test substances.
  • Fig. 10B shows the results of changes in the volume of xenograft tumors in individual A375 tumor-bearing mice resulting from the administration of exemplary antibodies of the present application and other test substances.
  • Figure 10C shows the results of the administration of exemplary antibodies of the present application and other test substances on the body weight of A375 tumor-bearing mice.
  • Figure 11A shows the results of the volume change of xenograft tumors in CT26 tumor-bearing mice produced by administration of exemplary humanized antibodies of the present application and other test substances.
  • Fig. 11B shows the results of the change of xenograft tumor volume in individual CT26 tumor-bearing mice produced by administration of exemplary humanized antibodies of the present application and other test substances.
  • Figure 11C shows the results of administration of exemplary humanized antibodies of the present application and other test substances on the body weight of CT26 tumor-bearing mice.
  • the term "subject” or “individual” refers to mammals, such as humans, but can also be other animals, such as wild animals, domestic animals or experimental animals (such as orangutans, monkeys, rats, mice, rabbits, guinea pigs, etc.) , marmots, ground squirrels, etc.).
  • antigen is a predetermined target to which an antibody can selectively bind.
  • antigens include, but are not limited to, polypeptides, sugars, nucleic acids, lipids, haptens, or other naturally occurring or synthetic compounds.
  • an “antibody” may refer to an immunoglobulin molecule capable of specifically binding to a target via at least one antigen recognition site located in the variable region of the immunoglobulin molecule, thus encompassing intact/full-length antibodies, antibody monoclonal chain or any antigen-binding fragment of an antibody (also referred to as an "antigen-binding portion").
  • antibody and “antigen-binding fragment/antigen-binding part” appear in the same context, “antibody” can be understood as a complete body relative to "antigen-binding fragment/antigen-binding part", and the two together correspond to the generalized Antibody concept.
  • anti-CD73 antibody or "antibody that binds CD73” includes antibodies that are capable of binding CD73 with sufficient affinity such that the antibody is useful as a diagnostic and/or therapeutic agent when targeted to CD73.
  • the anti-CD73 antibody binds to an irrelevant non-CD73 protein to an extent that is less than about 10% of the binding of the antibody to CD73, such as by fluorescence activated cell sorting (FACS) analysis or an immunoassay (e.g., radioimmunoassay (RIA)) assay.
  • FACS fluorescence activated cell sorting
  • an immunoassay e.g., radioimmunoassay (RIA)
  • the antibody that binds CD73 has a dissociation constant (KD) of less than or equal to 500 nM, 100 nM, 10 nM, 9 nM, 8 nM , 7nM, 6nM, 5nM, 4nM, 0.9nM, 0.8nM, 0.7nM, 0.6nM, 0.5nM, 0.4nM, 0.3nM, 0.2nM or 0.1nM.
  • KD dissociation constant
  • the anti-CD73 antibody binds to an epitope of the CD73 protein that is conserved between CD73 from different species (eg, between human and cynomolgus CD73).
  • agonist antibody is an antibody that elicits a response, eg, an antibody that mimics at least one functional activity of a polypeptide of interest.
  • Agonist antibodies include antibodies that are ligand mimics, e.g., wherein the ligand binds to a cell surface receptor, the binding induces cell signaling or activity through an intracellular cell signaling pathway, and wherein the antibody induces similar cellular Signal transduction or activation.
  • Full-length antibody refers to a protein comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds.
  • Each heavy chain comprises a heavy chain variable region (abbreviated VH) and a heavy chain constant region.
  • the heavy chain constant region comprises three domains, CH1, CH2 and CH3.
  • Each light chain comprises a light chain variable region (abbreviated VL) and a light chain constant region.
  • the light chain constant region comprises one domain, CL.
  • the VH and VL regions can be subdivided into regions of high variability called complementarity determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs).
  • CDRs complementarity determining regions
  • Each VH and VL consists of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.
  • These variable regions of the heavy and light chains contain the binding domains that interact with the antigen.
  • the constant regions of the antibodies can mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (such as effector cells) and the first component (Clq) of the classical complement system.
  • a full-length antibody may be of any class of antibody, such as IgD, IgE, IgG, IgA, or IgM (or subclasses of the above), but the antibody need not belong to any particular class.
  • immunoglobulins can be assigned to different classes.
  • immunoglobulins there are five main classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these classes can be further divided into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2.
  • the heavy chain constant domains corresponding to the different classes of immunoglobulins are called ⁇ , ⁇ , ⁇ , ⁇ , and ⁇ .
  • the subunit structures and three-dimensional structures of the different classes of immunoglobulins are well known. Chimeric or humanized antibodies are also encompassed by antibodies according to the present application.
  • complementarity determining regions are regions in the variable region that have the greatest influence on the affinity and specificity of antibodies.
  • CDR amino acid sequence of VH or VL There are many common ways to define the CDR amino acid sequence of VH or VL, such as Kabat definition, IMGT definition, Chothia definition, etc.
  • the CDR amino acid sequences in the VH and VL amino acid sequences can usually be determined according to different definition methods. In embodiments of the present application, Kabat sense CDR amino acid sequences are utilized.
  • the amino acid sequences of the CDRs in the variable region amino acid sequence can be analyzed in a number of ways.
  • mouse antibody refers to an antibody that is derived from the fusion of B cells and myeloma cells of immunized mice, and the mouse hybrid fusion cells that can proliferate indefinitely and secrete antibodies are screened, and then screened, The antibodies obtained by preparation and purification, murine antibodies are generally immunogenic, so they need to be humanized later.
  • humanized antibody means an antibody obtained by grafting CDR sequences derived from the germline of another mammalian species, such as a mouse, onto human framework sequences. To preserve binding affinity, some residues of the backbone (termed FR) segments can be modified. Humanized antibodies or fragments thereof according to the present application can be prepared by techniques known to those skilled in the art;
  • chimeric antibody refers to an antibody in which the variable region sequences are from one species and the constant region sequences are from another species, eg, an antibody in which the variable region sequences are from a mouse antibody and the constant region sequences are from a human antibody.
  • the chimeric antibody or fragment thereof according to the present application can be produced by using gene recombination technology.
  • said chimeric antibodies can be produced by cloning recombinant DNA comprising a promoter and sequences encoding the variable regions of non-human, especially murine monoclonal antibodies according to the present application, and sequences encoding the constant regions of human antibodies .
  • the chimeric antibody of the present application encoded by such a recombinant gene will be, for example, a mouse-human chimera whose specificity is determined by a variable region derived from mouse DNA and whose isotype is determined by a variable region derived from human DNA. to determine the constant region.
  • partially humanized antibody refers to an antibody that contains constant regions from a human and variable regions (including CDRs) from a non-human (eg, murine).
  • si-humanized antibody is a type of humanized antibody, and refers to an antibody in which one antibody chain contains a murine variable region and the other antibody chain contains a humanized variable region, that is, a semi-humanized antibody .
  • the term "monoclonal antibody” refers to an antibody obtained from a population of substantially homogeneous antibodies, ie, the individual antibodies comprising the population are identical except for the possibility of naturally occurring mutations in a small number of individuals.
  • antigen-binding fragment or “antigen-binding portion” or “antigen-binding region” is used interchangeably and refers to a protein comprising amino acid residues that interact with an antigen and confer on the binding agent its specificity and affinity for the antigen.
  • an antibody fragment such as Fv, Fab, F(ab') 2 or Fab', or any fragment that should be able to increase the half-life by chemical modification or by incorporation into liposomes
  • said chemical Modifications such as addition of poly(alkylene) glycols such as polyethylene glycol (“pegylated, PEGylated”) (known as Fv-PEG, scFv-PEG, Fab-PEG, F(ab') 2 - PEG or a pegylated fragment of Fab'-PEG) (“PEG” is polyethylene glycol), which fragment has CD73 binding activity.
  • the functional fragment will consist of or comprise a partial sequence of the variable chain of the heavy or light chain of the antibody from which it is derived, said partial sequence being sufficient to retain the same binding specificity and sufficient affinity as the antibody from which it is derived.
  • Fragments will contain a minimum of 5 amino acids, preferably 10, 15, 25, 50 and 100 contiguous amino acids of the antibody sequence from which they are derived.
  • antigen-binding fragments include, but are not limited to: (1) Fab fragments, which may be monovalent fragments having a VL-CL chain and a VH-CH1 chain; ( 2 ) F(ab') fragments, which may be monovalent fragments having two A bivalent fragment of a Fab' fragment, the two Fab' fragments are connected by a disulfide bridge in the hinge region (ie a dimer of Fab'); (3) an Fv fragment with the VL and VH domains of a single arm of an antibody.
  • single-chain antibody refers to a single polypeptide chain that is connected by a VH domain and a VL domain via a peptide linker.
  • (scFv) 2 comprises two VH domains and two VL domains connected by a peptide linker, the two The VL domain is combined with the two VH domains via a disulfide bridge.
  • Fc fragment refers to a portion of the constant region of an antibody heavy chain, including the hinge, CH2 and CH3 fragments of the constant region .
  • the Fc region of an anti-CD73 antibody can be engineered or modified, including effector function-related modifications, such as to reduce or eliminate antibody-dependent cellular Toxicity (ADCC) and/or complement-dependent cytotoxicity (CDC), which can be achieved by introducing one or more amino acid substitutions/mutations in the Fc region of the antibody.
  • ADCC antibody-dependent cellular Toxicity
  • CDC complement-dependent cytotoxicity
  • binding refers to a non-random binding reaction between two molecules, such as the binding of an antibody to an antigenic epitope.
  • polyantibody also known as “multispecific antibody” is a molecule with binding specificities for at least two different antigens, among which, such molecules that only bind to two antigens are also called double antibodies (ie, bispecific antibodies).
  • Antibodies, BsAb are also known as “multispecific antibody”.
  • bispecific antibody refers to an antibody that has the ability to bind two epitopes at the same time.
  • the two epitopes can be on different antigens or on the same antigen.
  • Bispecific antibodies can have a variety of structural configurations.
  • a bispecific antibody can be composed of two Fc fragments and two antigen-binding parts fused to them respectively (similar to natural antibodies, except that the two arms bind different antigenic targets or epitopes), and the antigen-binding parts can be single Chain antibody (scfv) format or Fab fragment format.
  • a human antibody that specifically binds CD73 can include at most about 1, at most about 2, at most about 5, at most about 10, or at most about 15 conservative substitutions and specifically binds a CD73 polypeptide.
  • conservative substitutions also includes the use of substituted amino acids in place of the unsubstituted parent amino acid, so long as the antibody specifically binds CD73. Non-conservative substitutions are those that reduce activity or bind to CD73.
  • XaaaY such as L234A, L235A, etc.
  • aaa represents the sequence position of an amino acid or base
  • aaa represents the residue sequence position in the reference sequence; or it can also follow the position numbering method prevailing in the field, such as the EU numbering system, etc.
  • X represents the original amino acid or base at "aaa”
  • Y represents the changed amino acid or base at "aaa”.
  • L234A mutation of the human heavy chain constant region it means that the 234th position leucine (L) is mutated to alanine (A) according to the EU numbering system of the human heavy chain constant region.
  • drugs A and B can be administered separately in sequence, administered separately at the same time, formulated in the same drug and administered together.
  • drugs A and B can be prepared in the same composition, or in the same pharmaceutical kit, at the time of manufacture.
  • Drugs A and B can be physically separate, or joined together.
  • the administration of drugs A and B may be separate, simultaneous or overlapping in time.
  • isolated refers to a biological component (such as a nucleic acid, protein (including antibodies) or organelle) that has been separated from other biological components (ie, other chromosomal and extrachromosomal DNA and RNA, proteins, and organelles) are substantially isolated or purified.
  • Nucleic acids and proteins that have been "isolated” include nucleic acids and proteins purified using standard purification methods.
  • the term also includes nucleic acids and proteins produced by recombinant expression in host cells as well as chemically synthesized nucleic acids.
  • derivative sequence refers to a sequence that has at least 80% (preferably 85%, 90%, 95%, 98% or 99%) sequence identity to a related sequence and still has the same or similar function.
  • pharmaceutical composition refers to a combination of at least one drug and optionally a pharmaceutically acceptable carrier or adjuvant combined together to achieve a specific purpose.
  • the pharmaceutical composition includes combinations that are separated in time and/or space, as long as they can work together to achieve the purpose of the present application.
  • the components contained in the pharmaceutical composition eg, antibodies, nucleic acid molecules, nucleic acid molecule combinations and/or conjugates according to the present application
  • the ingredients contained in the pharmaceutical composition are administered to the individual separately, the ingredients may be administered to the individual simultaneously or sequentially.
  • the pharmaceutically acceptable carrier is water, buffered aqueous solution, isotonic Saline solutions such as PBS (phosphate buffered saline), dextrose, mannitol, dextrose, lactose, starch, magnesium stearate, cellulose, magnesium carbonate, 0.3% glycerin, hyaluronic acid, ethanol, or polyalkylene diacetates Alcohols such as polypropylene glycol, triglycerides, etc.
  • the type of pharmaceutically acceptable carrier used depends inter alia on whether the composition according to the present application is formulated for oral, nasal, intradermal, subcutaneous, intramuscular or intravenous administration.
  • compositions according to the present application may comprise wetting agents, emulsifiers or buffer substances as additives.
  • the pharmaceutical composition or pharmaceutical preparation according to the present application can be administered by any suitable route, for example, it can be administered orally, nasally, intradermally, subcutaneously, intramuscularly or intravenously.
  • the term "therapeutically effective amount” or “effective amount” refers to a dose sufficient to demonstrate benefit to the subject to which it is administered.
  • the actual amount administered, as well as the rate and time course of administration, will depend upon the individual condition and severity of the individual being treated.
  • the prescribing of treatment e.g. decisions on dosage etc.
  • the prescribing of treatment is ultimately the responsibility of and relies on the general practitioner and other medical practitioners to make decisions, usually taking into account the disease being treated, the individual patient's condition, the site of delivery, the method of administration and what is known to the physician. other known factors.
  • the EC 50 value mainly refers to the concentration of a drug, antibody or toxin that can achieve 50% of the maximum biological effect after a specific exposure time.
  • EC 50 is also used to characterize the potency (including agonism and antagonism) of a compound at the cellular level, and the EC 50 value can be determined by methods such as ELISA.
  • fusion protein is a protein composed of at least two structural domains, each domain is not naturally associated, encoded by a separate gene, each gene is linked, and is recognized as a whole. Transcription and translation produce a single protein.
  • a "fusion protein” comprising an antibody or an antigen-binding part refers to a product obtained by fusing an antibody or an antigen-binding part with another biologically active protein using genetic engineering techniques. Antigen binding ability and unique biological properties of bioactive proteins fused to antibodies.
  • identity/homology/identity with respect to amino acid or nucleic acid sequences is defined as the percentage of residues in amino acid or nucleotide sequence variants that are identical after alignment of the sequences and introduction of gaps, if desired, to Maximum percent identity. Methods and computer programs for alignment are well known in the art.
  • tumor refers to a neoplasm or solid lesion formed by abnormal cell growth. Tumors can be benign, pre-malignant, or malignant.
  • malignancy refers to or describes the physiological condition in mammals that is typically characterized by unregulated cell growth.
  • exemplary malignancies include: carcinomas, solid tumors, melanoma sarcomas, hematological tumors, germ cell tumors, and blastomas.
  • malignancies include: multiple myeloma, renal cancer, lung cancer including small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, and lung squamous cell carcinoma, bladder cancer, breast cancer, cervical cancer, colon cancer, including Gastrointestinal cancer of stomach, prostate, pancreas, peritoneum, hepatocellular carcinoma, glioblastoma, ovary, liver, urinary tract, hepatoma, rectum, colorectum, endometrium or uterus Carcinoma, salivary gland cancer, squamous cell carcinoma (eg, squamous cell carcinoma), vulvar cancer, thyroid cancer, anal cancer, penile cancer, melanoma and B-cell lymphoma, brain and head and neck cancer and associated metastases.
  • lung cancer including small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, and lung squamous cell carcinoma
  • bladder cancer breast cancer, cervical cancer, colon cancer
  • blood cancer refers to the uncontrolled growth and proliferation of abnormal cells, which in most cases originate from the bone marrow, which is where blood cells are produced.
  • exemplary hematological tumors include various types of leukemia, multiple myeloma, and malignant lymphoma.
  • blood cancers include: acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), hairy cell leukemia (HCL), T-cell prolymphocytic leukemia, large granular lymphocytic leukemia, juvenile myelomonocytic leukemia, B-cell prolymphocytic leukemia, Burkitt's leukemia and adult T-cell leukemia, non-Hodgkin Lymphoma, B-cell lymphoma, small lymphocytic lymphoma, lymphoplasmacytic lymphoma, primary macroglobulinemia ( macroglobulinemia), splenic marginal zone lymphoma, plasmacytoma, extranodal marginal zone B-cell lymphoma, MALT lymphoma, nodal marginal zone B-cell lymphoma (NMZL), follicular lymphoma
  • ALL
  • solid tumor refers to a tangible mass that can be palpated by clinical examination such as X-ray film, CT scan, B-ultrasound or palpation.
  • Clinically diagnosed and treated solid tumors are divided into malignant and benign.
  • Malignant solid tumors include: Pediatric Hodgkin's lymphoma: lymphocyte predominant, nodular sclerosis, mixed cellularity, lymphocytic depletion; Pediatric non-Hodgkin's lymphoma: prolymphoblastic lymphoma, small noncleaved Cell lymphoma (Burkitt/non-Burkitt lymphoma), diffuse large B-cell lymphoma, anaplastic large cell lymphoma, etc.; renal tumors in children: Wilms tumor (Wilms tumor), clear cell renal cell carcinoma, Renal rhabdoid tumor, renal clear cell sarcoma, renal primitive neuroectodermal tumor, etc.; neuroblastoma in children: neuroblastoma, ganglion
  • the application provides an anti-CD73 antibody or an antigen-binding portion thereof, the anti-CD73 antibody or an antigen-binding portion thereof comprising a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising a heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3), the light chain variable region includes light chain CDR1 (LCDR1), light chain CDR2 (LCDR2) and light chain CDR3 (LCDR3), where:
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1
  • the sequence of HCDR2 is SEQ ID NO.2
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or with the sequence shown in SEQ ID NO.
  • sequence shown in 3 has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR1 is the sequence shown in SEQ ID NO.5 or has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR2 is the sequence shown in SEQ ID NO.6 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.6 99% identity
  • sequence of LCDR3 is the sequence shown in SEQ ID NO.7 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.7; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1, and the sequence of HCDR2 is SEQ ID NO.17
  • the sequence shown is at least 80%, 85%, 90%, 95% or 99% identical to the sequence shown in SEQ ID NO.17
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or the same as SEQ ID NO.
  • sequence shown in 3 has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR1 is the sequence shown in SEQ ID NO.5 or has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR2 is the sequence shown in SEQ ID NO.32 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.32 99% identity
  • sequence of LCDR3 is the sequence shown in SEQ ID NO.7 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.7; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1
  • the sequence of HCDR2 is SEQ ID NO.17
  • the sequence shown may have at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.17
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or with the sequence shown in SEQ ID NO.
  • the sequence shown in 3 has at least 80%, 85%, 90%, 95% or 99% identity
  • the sequence of LCDR1 is the sequence shown in SEQ ID NO.5 or has at least 80%, 85%, 90%, 95% or 99% identity
  • the sequence of LCDR2 is the sequence shown in SEQ ID NO.6 or with SEQ ID NO.6
  • the sequence shown has at least 80%, 85%, 90%, 95% or 99% identity
  • the sequence of LCDR3 is the sequence shown in SEQ ID NO.7 or has at least 80%, 85% identity with the sequence shown in SEQ ID NO.7 %, 90%, 95% or 99% identity; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1
  • the sequence of HCDR2 is SEQ ID NO.2
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or with the sequence shown in SEQ ID NO.
  • sequence shown in 3 has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR1 is the sequence shown in SEQ ID NO.5 or has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR2 is the sequence shown in SEQ ID NO.32 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.32 99% identity
  • sequence of LCDR3 is the sequence shown in SEQ ID NO.7 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.7; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1
  • the sequence of HCDR2 is SEQ ID NO.21
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or with the sequence shown in SEQ ID NO.
  • sequence shown in 3 has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR1 is the sequence shown in SEQ ID NO.5 or has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR2 is the sequence shown in SEQ ID NO.32 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.32 99% identity
  • sequence of LCDR3 is the sequence shown in SEQ ID NO.7 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.7; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1
  • the sequence of HCDR2 is SEQ ID NO.21
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or with the sequence shown in SEQ ID NO.
  • sequence shown in 3 has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR1 is the sequence shown in SEQ ID NO.5 or has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR2 is the sequence shown in SEQ ID NO.6 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.6 99% identity
  • sequence of LCDR3 is the sequence shown in SEQ ID NO.7 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.7; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1
  • the sequence of HCDR2 is SEQ ID NO.23
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or with the sequence shown in SEQ ID NO.
  • sequence shown in 3 has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR1 is the sequence shown in SEQ ID NO.5 or has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR2 is the sequence shown in SEQ ID NO.32 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.32 99% identity
  • sequence of LCDR3 is the sequence shown in SEQ ID NO.7 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.7; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1
  • the sequence of HCDR2 is SEQ ID NO.23
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or with the sequence shown in SEQ ID NO.
  • sequence shown in 3 has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR1 is the sequence shown in SEQ ID NO.5 or has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR2 is the sequence shown in SEQ ID NO.6 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.6 99% identity
  • sequence of LCDR3 is the sequence shown in SEQ ID NO.7 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.7; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.9 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.9
  • the sequence of HCDR2 is SEQ ID NO.10
  • the sequence shown or with the SEQ ID has at least 80%, 85%, 90%, 95% or 99% identity
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.11 or has at least 80% identity with the sequence shown in SEQ ID NO.11 %, 85%, 90%, 95% or 99% identity
  • the sequence of LCDR1 is the sequence shown in SEQ ID NO.13 or has at least 80%, 85%, 90%, 95% with the sequence shown in SEQ ID NO.13 % or 99% identity
  • the sequence of LCDR2 is the sequence shown in SEQ ID NO.14 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.14
  • the sequence of LCDR3 The sequence is or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.9 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.9
  • the sequence of HCDR2 is SEQ ID NO.10
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.11 or with the sequence shown in SEQ ID NO.
  • the sequence shown in 11 has at least 80%, 85%, 90%, 95% or 99% identity
  • the sequence of LCDR1 is the sequence shown in SEQ ID NO.35 or has at least 80%, 85%, 90%, 95% or 99% identity
  • the sequence of LCDR2 is the sequence shown in SEQ ID NO.14 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.14 99% identity
  • the sequence of LCDR3 is the sequence shown in SEQ ID NO.15 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.15; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.9 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.9
  • the sequence of HCDR2 is SEQ ID NO.27
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.11 or with the sequence shown in SEQ ID NO.
  • the sequence shown in 11 has at least 80%, 85%, 90%, 95% or 99% identity
  • the sequence of LCDR1 is the sequence shown in SEQ ID NO.35 or has at least 80%, 85%, 90%, 95% or 99% identity
  • the sequence of LCDR2 is the sequence shown in SEQ ID NO.14 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.14 99% identity
  • the sequence of LCDR3 is the sequence shown in SEQ ID NO.15 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.15; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.9 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.9
  • the sequence of HCDR2 is SEQ ID NO.27
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.11 or with the sequence shown in SEQ ID NO.
  • sequence shown in 11 has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR1 is the sequence shown in SEQ ID NO.13 or has at least 80%, 85%, 90%, 95% or 99% identity
  • sequence of LCDR2 is the sequence shown in SEQ ID NO.14 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.14 99% identity
  • sequence of LCDR3 is the sequence shown in SEQ ID NO.15 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.15; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.9 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.9
  • the sequence of HCDR2 is SEQ ID NO.30
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.11 or with the sequence shown in SEQ ID NO.
  • the sequence shown in 11 has at least 80%, 85%, 90%, 95% or 99% identity
  • the sequence of LCDR1 is the sequence shown in SEQ ID NO.35 or has at least 80%, 85%, 90%, 95% or 99% identity
  • the sequence of LCDR2 is the sequence shown in SEQ ID NO.14 or has at least 80%, 85%, 90%, 95% or the sequence shown in SEQ ID NO.14 99% identity
  • the sequence of LCDR3 is the sequence shown in SEQ ID NO.15 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.15; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.9 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.9
  • the sequence of HCDR2 is SEQ ID NO.30
  • the sequence of HCDR3 is SEQ ID
  • the sequence of LCDR1 is the sequence shown in SEQ ID NO.13 or with SEQ ID NO.13
  • the sequence shown in ID NO.13 has at least 80%, 85%, 90%, 95% or 99% identity
  • the sequence of LCDR2 is the sequence shown in SEQ ID NO.14 or has at least the same sequence with the sequence shown in SEQ ID NO.14 80%, 85%, 90%, 95% or 99% identity
  • HCDR and LCDR sequences are defined according to Kabat.
  • the heavy chain variable region has the amino acid sequence shown in SEQ ID NO.4, 18, 19, 20, 22, 24, 12, 25, 26, 28, 29 or 31 or the same , 18, 19, 20, 22, 24, 12, 25, 26, 28, 29 or 31 amino acid sequences having at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identity sequence.
  • the light chain variable region has the amino acid sequence shown in SEQ ID NO.8, 33, 34, 16, 36, 37 or 38 or the or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity to the amino acid sequence shown in 38.
  • the heavy chain variable region sequence is the sequence shown in SEQ ID NO.4, and the light chain variable region sequence is the sequence shown in SEQ ID NO.8; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.18, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.33; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.18, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.34; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.19, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.33; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.19, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.34; or
  • the heavy chain variable region sequence is the sequence shown in SEQ ID NO.20, and the light chain variable region sequence is the sequence shown in SEQ ID NO.33; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.20, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.34; or
  • the heavy chain variable region sequence is the sequence shown in SEQ ID NO.22, and the light chain variable region sequence is the sequence shown in SEQ ID NO.33; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.22, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.34; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.24, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.33; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.24, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.34; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.12, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.16; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.25, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.36; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.25, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.37; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.25, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.38; or
  • the heavy chain variable region sequence is the sequence shown in SEQ ID NO.26, and the light chain variable region sequence is the sequence shown in SEQ ID NO.36; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.26, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.37; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.26, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.38; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.28, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.36; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.28, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.37; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.28, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.38; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.29, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.36; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.29, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.37; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.29, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.38; or
  • the sequence of the heavy chain variable region is the sequence shown in SEQ ID NO.31, and the sequence of the light chain variable region is the sequence shown in SEQ ID NO.36; or
  • sequence of the variable region of the heavy chain is the sequence shown in SEQ ID NO.31, and the sequence of the variable region of the light chain is the sequence shown in SEQ ID NO.37; or
  • the sequence of the variable region of the heavy chain is the sequence shown in SEQ ID NO.31, and the sequence of the variable region of the light chain is the sequence shown in SEQ ID NO.38.
  • the present application provides an anti-CD73 antibody or an antigen-binding portion thereof, the anti-CD73 antibody or an antigen-binding portion thereof comprising a heavy chain variable region and a light chain variable region, wherein:
  • the heavy chain variable region has an amino acid sequence shown in SEQ ID NO.4, 18, 19, 20, 22, 24, 12, 25, 26, 28, 29 or 31 or the same as SEQ ID NO.4, 18, 19 , 20, 22, 24, 12, 25, 26, 28, 29 or 31 have an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to the amino acid sequence; and/ or
  • the light chain variable region has an amino acid sequence shown in SEQ ID NO.8, 33, 34, 16, 36, 37 or 38 or the same as shown in SEQ ID NO.8, 33, 34, 16, 36, 37 or 38 Amino acid sequences are amino acid sequences that are at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical.
  • the present application provides an anti-CD73 antibody or an antigen-binding portion thereof, the anti-CD73 antibody or an antigen-binding portion thereof comprising a heavy chain variable region, the heavy chain variable region comprising a heavy chain CDR1 (HCDR1), a heavy chain chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3), wherein:
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1
  • the sequence of HCDR2 is SEQ ID
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or with SEQ ID NO.
  • the sequence shown in ID NO.3 has at least 80%, 85%, 90%, 95% or 99% identity; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1
  • the sequence of HCDR2 is SEQ ID
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or with SEQ ID NO.3
  • the sequence shown in ID NO.3 has at least 80%, 85%, 90%, 95% or 99% identity; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1, and the sequence of HCDR2 is SEQ ID The sequence shown in NO.21 or with SEQ The sequence shown in ID NO.21 has at least 80%, 85%, 90%, 95% or 99% identity, and the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or has at least the same sequence with the sequence shown in SEQ ID NO.3 80%, 85%, 90%, 95% or 99% identity; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.1 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.1
  • the sequence of HCDR2 is SEQ ID
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.3 or with SEQ ID NO.3
  • the sequence shown in ID NO.3 has at least 80%, 85%, 90%, 95% or 99% identity; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.9 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.9
  • the sequence of HCDR2 is SEQ ID
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.11 or with the sequence shown in SEQ ID NO.10
  • the sequence shown in ID NO.11 has at least 80%, 85%, 90%, 95% or 99% identity; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.9 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.9
  • the sequence of HCDR2 is SEQ ID
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.11 or with the sequence shown in SEQ ID NO.11
  • the sequence shown in ID NO.11 has at least 80%, 85%, 90%, 95% or 99% identity; or
  • the sequence of HCDR1 is the sequence shown in SEQ ID NO.9 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.9
  • the sequence of HCDR2 is SEQ ID
  • the sequence of HCDR3 is the sequence shown in SEQ ID NO.11 or with SEQ ID NO.11
  • the sequence shown in ID NO.11 has at least 80%, 85%, 90%, 95% or 99% identity;
  • HCDR sequence is defined according to Kabat.
  • the present application provides an anti-CD73 antibody or an antigen-binding portion thereof, the anti-CD73 antibody or an antigen-binding portion thereof comprising a light chain variable region, the light chain variable region comprising a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2) and light chain CDR3 (LCDR3), wherein:
  • the sequence of LCDR1 is the sequence shown in SEQ ID NO.5 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.5, and the sequence of LCDR2 is SEQ ID The sequence shown in NO.6 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.6, the sequence of LCDR3 is the sequence shown in SEQ ID NO.7 or with SEQ ID NO. The sequence shown in ID NO.7 has at least 80%, 85%, 90%, 95% or 99% identity; or
  • the sequence of LCDR1 is the sequence shown in SEQ ID NO.5 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.5, and the sequence of LCDR2 is SEQ ID The sequence shown in NO.32 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.32, and the sequence of LCDR3 is the sequence shown in SEQ ID NO.7 or with SEQ ID NO.7 The sequence shown in ID NO.7 has at least 80%, 85%, 90%, 95% or 99% identity; or
  • the sequence of LCDR1 is the sequence shown in SEQ ID NO.13 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.13
  • the sequence of LCDR2 is SEQ ID
  • the sequence of LCDR3 is the sequence shown in SEQ ID NO.15 or with SEQ ID NO.14
  • the sequence shown in ID NO.15 has at least 80%, 85%, 90%, 95% or 99% identity; or
  • the sequence of LCDR1 is the sequence shown in SEQ ID NO.35 or has at least 80%, 85%, 90%, 95% or 99% identity with the sequence shown in SEQ ID NO.35
  • the sequence of LCDR2 is SEQ ID
  • the sequence of LCDR3 is the sequence shown in SEQ ID NO.15 or with SEQ ID NO.15
  • the sequence shown in ID NO.15 has at least 80%, 85%, 90%, 95% or 99% identity;
  • LCDR sequence is defined according to Kabat.
  • the anti-CD73 antibody, or antigen-binding portion thereof binds human CD73. In some embodiments, the anti-CD73 antibody or antigen-binding portion thereof binds to an epitope of the CD73 protein that is conserved between CD73 from different species (eg, between human and cynomolgus CD73).
  • the anti-CD73 antibody, or antigen-binding portion thereof has ADCC activity.
  • the anti-CD73 antibody, or antigen-binding portion thereof has CDC activity.
  • the anti-CD73 antibody is a whole antibody, a single chain antibody (scFv), scFv-Fc, a bispecific antibody or a multispecific antibody.
  • the antigen binding portion of the anti-CD73 antibody is Fab, Fab', Fv or F(ab') 2 .
  • the anti-CD73 antibody is a murine antibody, a chimeric antibody, a humanized antibody or a fully human antibody.
  • the anti-CD73 antibody is a monoclonal antibody.
  • the anti-CD73 antibody is an antibody of the IgM, IgD, IgG, IgA or IgE type. In some embodiments, the anti-CD73 antibody is an IgG antibody. In some more specific embodiments, the anti-CD73 antibody is an IgG1 antibody.
  • the anti-CD73 antibody is of the IgGl, IgG2 or IgG4 isotype.
  • the anti-CD73 antibody comprises a light chain constant region of the kappa subtype or the lambda subtype.
  • the anti-CD73 antibody comprises a human IgGl heavy chain constant region and a human kappa light chain constant region.
  • the anti-CD73 antibody or antigen-binding portion thereof is used in medical therapy.
  • the anti-CD73 antibody comprises a wild-type Fc region.
  • the anti-CD73 antibody, or antigen-binding portion thereof is engineered to have enhanced ADCC or CDC activity.
  • an anti-CD73 antibody comprises an engineered Fc region that renders the antibody having enhanced ADCC and/or CDC effects.
  • the anti-CD73 antibody, or antigen-binding portion thereof is engineered to have reduced ADCC or CDC activity.
  • the anti-CD73 antibody comprises an engineered Fc region such that the antibody has reduced ADCC and/or CDC effects.
  • antibodies with weakened or even eliminated ADCC or CDC activity may be beneficial, because too strong Fc effect may affect or even weaken the therapeutic effect of antibodies. Ways to weaken or eliminate Fc effects are known in the art, for example, 1) amino acid residue mutation modification (mainly weakening the binding to related receptors), 2) glycosylation modification, 3) IgG4 antibody modification, etc.
  • the antibody has a human IgG1 heavy chain constant region and contains mutations at one or more of L234, L235, P329, P331, wherein residue numbering follows the EU numbering system, e.g., the mutation is L234A One or more mutations in , L235A, P329A, P331S.
  • the antibody has a heavy chain variable region shown in SEQ ID NO.20, a light chain variable region shown in SEQ ID NO.33, and a light chain variable region shown in SEQ ID NO.39 The heavy chain constant region shown and the light chain constant region shown in SEQ ID NO.40.
  • the antibody has a heavy chain variable region shown in SEQ ID NO.31, a light chain variable region shown in SEQ ID NO.38, and a light chain variable region shown in SEQ ID NO.39 Shown heavy chain constant region and SEQ ID NO.40 Light chain constant region indicated.
  • the present application provides an isolated nucleic acid molecule encoding the anti-CD73 antibody or antigen-binding portion thereof described in the first to fourth aspects.
  • the application provides a combination of isolated polynucleotides comprising a polynucleotide encoding a light chain of an antibody of the application or an antigen-binding portion thereof and a heavy chain encoding an antibody of the application or an antigen-binding portion thereof chain of polynucleotides.
  • the polynucleotide is operably linked to a regulatory sequence that is recognized by a host cell transformed with the vector.
  • the present application provides a vector (such as an expression vector), which contains the nucleic acid molecule or polynucleotide combination described in the fifth aspect.
  • the expression vector of the present application comprises the nucleic acid molecule described in the present application or the combination of the polynucleotide described in the present application, and described polynucleotide and the polypeptide that allow its coding express in host cell or cell-free
  • the regulatory sequences expressed in the system are operably linked.
  • the choice of expression vector depends on the choice of the host cell and can be selected so as to have the desired expression and regulatory characteristics in the host cell of choice.
  • An "expression vector” is a vector that includes one or more expression control sequences, which are DNA sequences that control and regulate the transcription and/or translation of another DNA sequence.
  • the nucleic acid in the vector may be operably linked to one or more expression control sequences.
  • "operably linked” means incorporated into a genetic construct such that expression control sequences effectively control the expression of a coding sequence of interest.
  • expression control sequences include promoters, enhancers, and transcription termination regions.
  • a promoter is an expression control sequence consisting of a region of a DNA molecule usually within 100 nucleotides upstream of the start of transcription (usually near the initiation site for RNA polymerase II).
  • the translation initiation site of the translational reading frame of the polypeptide must be positioned between 1 and about 50 nucleotides downstream of the promoter.
  • Enhancers provide expression specificity in terms of time, location and level. Unlike promoters, enhancers can function when located at different distances from the transcription site. Enhancers can also be located downstream of the transcription initiation site.
  • a coding sequence is "operably linked to” and “under the control of” an expression control sequence in a cell when RNA polymerase is capable of transcribing the coding sequence into mRNA, which can then be translated into the protein encoded by the coding sequence ".
  • Suitable expression vectors include, but are not limited to, plasmid and viral vectors derived from, for example, bacteriophage, baculovirus, tobacco mosaic virus, herpes virus, cytomegalovirus, retrovirus, vaccinia virus, adenovirus, and adeno-associated virus.
  • Many vectors and expression systems are commercially available from companies such as Novagen (Madison, WI), Clontech (Palo Alto, CA), Stratagene (LaJolla, CA), and Invitrogen Life Technologies (Carlsbad, CA).
  • Expression vectors can include tag sequences.
  • Tag sequences are usually expressed as fusions to the encoded polypeptide. Such tags can be inserted anywhere within the polypeptide, including the carboxyl or amino termini. Examples of useful tags include, but are not limited to, Fc fragments, polyhistidine, green fluorescent protein (GFP), glutathione S-transferase (GST), c-myc, hemagglutinin, FlagTM tags (Kodak, New Haven, CT), maltose E-binding protein and protein A.
  • a nucleic acid molecule encoding a CD73 fusion polypeptide is present in a vector containing nucleic acid encoding one or more domains of an Ig heavy chain constant region, e.g., corresponding to the C ⁇ 1 chain of a human immunoglobulin. Amino acid sequences of the hinge region, CH2 region and CH3 region (Fc fragment).
  • the present application provides a host cell containing the nucleic acid molecule of the fifth aspect or the vector of the sixth aspect.
  • the host cell can be a prokaryotic host cell, a eukaryotic host cell, or a phage.
  • the prokaryotic host cell can be Escherichia coli, Bacillus subtilis, Streptomyces or Proteus mirabilis, etc.
  • Eukaryotic host cells can be fungi such as Pichia pastoris, Saccharomyces cerevisiae, Schizosaccharomyces, Trichoderma, insect cells such as armyworm, plant cells such as tobacco, such as BHK cells, CHO cells, COS cells, Mammalian cells such as myeloma cells.
  • the host cells are preferably mammalian cells, more preferably BHK cells, CHO cells, NSO cells or COS cells.
  • the present application provides an antibody-drug conjugate, also known as an antibody-drug conjugate, which comprises the anti-CD73 antibody or its antigen-binding portion described in the first to fourth aspects coupled to a therapeutic agent .
  • the therapeutic agent is an antineoplastic drug, eg, a cytotoxic drug, an immunopotentiator, or a radioisotope.
  • the type of cytotoxic drug includes tubulin inhibitors (eg, alkaloids), DNA topoisomerase inhibitors, DNA damaging agents, antimetabolites, or antitumor antibiotics.
  • tubulin inhibitors eg, alkaloids
  • DNA topoisomerase inhibitors e.g., DNA damaging agents, antimetabolites, or antitumor antibiotics.
  • tubulin inhibitors include, but are not limited to, auristatin derivatives (such as MMAE (Monomethyl auristatin E), MMAF (Monomethyl auristatin F)) or maytansine alkaloid derivatives (such as DM1, DM4, Ansamitocin, Mertansine or dolastatin and its derivatives).
  • auristatin derivatives such as MMAE (Monomethyl auristatin E), MMAF (Monomethyl auristatin F)
  • maytansine alkaloid derivatives such as DM1, DM4, Ansamitocin, Mertansine or dolastatin and its derivatives.
  • the DNA topoisomerase inhibitor is a camptothecin analog or a DNA topoisomerase I inhibitor and derivatives thereof, e.g., DXD, SN38, irinotecan, irinotecan hydrochloride, Camptothecin, 9-aminocamptothecin, 9-nitrocamptothecin, 10-hydroxycamptothecin, 9-chloro-10-hydroxycamptothecin, 22-hydroxycamptothecin, topotecan, le Totecan, belotecan, exitecan, homosilatecan, 6,8-dibromo-2-methyl-3-[2-(D-xylopyranosylamino)benzene Base]-4(3H)-quinazolinone, 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-(2E)-2-acrylamide, 2 -Cyano-3-(3,4-dihydroxyphenyl)
  • DNA damaging agents include, but are not limited to, calicheamicins, duocarmycins, antramycin derivatives PBD (pyrrolobenzodiazepine, pyrrolobenzodiazepine ).
  • antimetabolites include, but are not limited to, methotrexate, 6-mercaptopurine, or 5-fluorouracil.
  • antitumor antibiotics include, but are not limited to, polypeptide antibiotics (such as actinomycin D or bleomycin) or anthraquinones (such as doxorubicin or mitoxantrone hydrochloride).
  • radioisotopes include, but are not limited to, 211 At, 131 I, 125 I, 90 Y, 186 Re, 188 Re, 153 Sm, 212 Bi, 32 P , 60 Co, or 177 Lu.
  • immune potentiating agents include, but are not limited to, levamisole, pidotimod, imiquimod, isoprinosine, polyinosine, or polyinosine glycosides.
  • an anti-CD73 antibody of the present application is covalently linked to a drug moiety via a linker.
  • the linker is a cleavable linker.
  • linkers are cleavable under intracellular conditions.
  • the linker is hydrolyzable at a pH of less than 5.5.
  • linkers are cleavable by intracellular proteases.
  • the linker is a cathepsin cleavable linker.
  • the linker comprises a dipeptide.
  • the dipeptide is valine (Val)-citrulline (Cit).
  • the antibody is attached to the linker through a cysteine sulfhydryl group of the antibody.
  • the antibody is linked to the linker via an amino group of the antibody, particularly an amino group of a glutamine residue.
  • linkers include mc-Val-Cit-pAB, mc-Val-Cit-pABC, mc-Val-Cit, NH2-(PEG)m-Val-Cit, NH2- (PEG)m-Val- Cit-pAB, wherein m is an integer from 1 to 8.
  • the linker is a non-cleavable linker, including without limitation, an acid-labile linker (e.g., a hydrazone linker), a disulfide bond-containing linker, a peptidase-sensitive linker (e.g., a peptide linker comprising an amino acid, such as valline acid and/or citrulline such as citrulline-valine or phenylalanine-lysine), photolabile linkers, dimethyl linkers, thioether linkers, or those designed to avoid multidrug transporter-mediated Resistant hydrophilic linkers, etc.
  • an acid-labile linker e.g., a hydrazone linker
  • a disulfide bond-containing linker e.g., a disulfide bond-containing linker
  • a peptidase-sensitive linker e.g., a peptide linker comprising an amino acid, such as valline acid and/or citrulline such
  • the present application provides a pharmaceutical composition, which comprises the anti-CD73 antibody or its antigen-binding portion described in the first aspect to the fourth aspect or the antibody-drug conjugate described in the eighth aspect, and a pharmaceutically acceptable carrier.
  • the pharmaceutical composition is used to treat a tumor in an individual.
  • the tumor is a tumor that expresses CD73 (CD73+) on the surface of tumor cells.
  • the tumor is a tumor that highly expresses CD73 (CD73 + ) on the surface of tumor cells.
  • a tumor that highly expresses CD73 (CD73 + ) means that at least 60% of the tumor cells in the tumor cell population express CD73.
  • a tumor that highly expresses CD73 (CD73 + ) means that at least 70% of the tumor cells in the tumor cell population express CD73.
  • a tumor that highly expresses CD73 (CD73 + ) means that at least 80% of the tumor cells in the tumor cell population express CD73.
  • a tumor that highly expresses CD73 (CD73 + ) means that at least 90% of the tumor cells in the tumor cell population express CD73.
  • a tumor that highly expresses CD73 means that at least 95% of the tumor cells in the tumor cell population express CD73. in some implementations In the scheme, a tumor highly expressing CD73 (CD73 + ) means that at least 98% of the tumor cells in the tumor cell population express CD73. In some embodiments, a tumor that highly expresses CD73 (CD73 + ) means that at least 99% of the tumor cells in the tumor cell population express CD73.
  • the tumor is a solid tumor.
  • the tumor is a hematological tumor.
  • the tumor is a malignant tumor.
  • the tumor is cancer
  • the tumor is selected from the group consisting of ovarian cancer, breast cancer, melanoma, colorectal cancer, gastric cancer, kidney cancer, prostate cancer, pancreatic cancer, esophageal cancer, head and neck cancer, thyroid cancer, lung cancer, bladder cancer, cervical cancer , colon cancer, liver cancer, peritoneal cancer, hepatocellular carcinoma, glioblastoma, urinary tract cancer, rectal cancer, endometrial cancer, uterine cancer, salivary gland cancer, squamous cell carcinoma, vulvar cancer, anal cancer, penile cancer , brain cancer, lymphoma (eg, B-cell lymphoma), leukemia, and metastases of the above tumors.
  • lymphoma eg, B-cell lymphoma
  • the pharmaceutical composition may also contain one or more of: lubricants, such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying agents; suspending agents; preservatives, Such as benzoic acid, sorbic acid and calcium propionate; sweeteners and/or flavoring agents, etc.
  • lubricants such as talc, magnesium stearate, and mineral oil
  • wetting agents such as talc, magnesium stearate, and mineral oil
  • emulsifying agents such as benzoic acid, sorbic acid and calcium propionate
  • preservatives such as benzoic acid, sorbic acid and calcium propionate
  • sweeteners and/or flavoring agents etc.
  • the pharmaceutical compositions of the present application can be formulated as tablets, pills, powders, lozenges, elixirs, suspensions, emulsions, solutions, syrups, suppositories or capsules and the like.
  • the pharmaceutical composition of the present application can be delivered by any physiologically acceptable mode of administration, including but not limited to: oral administration, parenteral administration, nasal administration, rectal administration Drugs, intraperitoneal administration, intravascular injection, subcutaneous administration, transdermal administration, inhalation administration, etc.
  • compositions for therapeutic use may be formulated in the form of lyophilized formulations or aqueous solutions by mixing reagents of desired purity with, as appropriate, pharmaceutically acceptable carriers, excipients, etc. storage.
  • the present application provides the anti-CD73 antibody or its antigen-binding portion described in the first to fourth aspects, the nucleic acid molecule described in the fifth aspect, the carrier described in the sixth aspect, and the anti-CD73 antibody described in the seventh aspect.
  • the tumor is a tumor that expresses CD73 (CD73+) on the surface of tumor cells.
  • the tumor is a tumor that highly expresses CD73 (CD73 + ) on the surface of tumor cells.
  • a tumor that highly expresses CD73 (CD73 + ) means that at least 60% of the tumor cells in the tumor cell population express CD73.
  • a tumor that highly expresses CD73 (CD73 + ) means that at least 70% of the tumor cells in the tumor cell population express CD73.
  • a tumor that highly expresses CD73 (CD73 + ) means that at least 80% of the tumor cells in the tumor cell population express CD73.
  • a tumor that highly expresses CD73 (CD73 + ) means that at least 90% of the tumor cells in the tumor cell population express CD73.
  • a tumor that highly expresses CD73 means that at least 95% of the tumor cells in the tumor cell population express CD73. In some embodiments, a tumor that highly expresses CD73 (CD73 + ) means that at least 98% of the tumor cells in the tumor cell population express CD73. In some embodiments, a tumor that highly expresses CD73 (CD73 + ) means that at least 99% of the tumor cells in the tumor cell population express CD73.
  • the tumor is a solid tumor.
  • the tumor is a hematological tumor.
  • the tumor is a malignant tumor.
  • the tumor is cancer
  • the tumor is selected from the group consisting of ovarian cancer, breast cancer, melanoma, colorectal cancer, gastric cancer, kidney cancer, prostate cancer, pancreatic cancer, esophageal cancer, head and neck cancer, thyroid cancer, lung cancer, bladder cancer, cervical cancer , colon cancer, liver cancer, peritoneal cancer, hepatocellular carcinoma, glioblastoma, urinary tract cancer, rectal cancer, endometrial cancer, uterine cancer, salivary gland cancer, squamous cell carcinoma, vulvar cancer, anal cancer, penile cancer , brain cancer, lymphoma (eg, B-cell lymphoma), leukemia, and metastases of the above tumors.
  • lymphoma eg, B-cell lymphoma
  • the present application provides the treatment of tumors in individuals, especially tumor cells expressing CD73 on the surface (CD73+) tumors, the method comprising administering to the individual a therapeutically effective amount of the anti-CD73 antibody or antigen-binding portion thereof described in the first to fourth aspects, or the antibody drug conjugate described in the eighth aspect Or the pharmaceutical composition described in the ninth aspect.
  • the methods of treating or preventing a disease, disorder or condition comprise exposing cells to an anti-CD73 antibody in vitro.
  • the tumor is a tumor that expresses CD73 (CD73+) on the surface of tumor cells.
  • the tumor is a tumor that highly expresses CD73 (CD73 + ) on the surface of tumor cells.
  • a tumor that highly expresses CD73 (CD73 + ) means that at least 60% of the tumor cells in the tumor cell population express CD73.
  • a tumor that highly expresses CD73 (CD73 + ) means that at least 70% of the tumor cells in the tumor cell population express CD73.
  • a tumor that highly expresses CD73 (CD73 + ) means that at least 80% of the tumor cells in the tumor cell population express CD73.
  • a tumor that highly expresses CD73 (CD73 + ) means that at least 90% of the tumor cells in the tumor cell population express CD73.
  • a tumor that highly expresses CD73 means that at least 95% of the tumor cells in the tumor cell population express CD73. In some embodiments, a tumor that highly expresses CD73 (CD73 + ) means that at least 98% of the tumor cells in the tumor cell population express CD73. In some embodiments, a tumor that highly expresses CD73 (CD73 + ) means that at least 99% of the tumor cells in the tumor cell population express CD73.
  • the tumor is a solid tumor.
  • the tumor is a hematological tumor.
  • the tumor is a malignant tumor.
  • the tumor is cancer
  • the tumor is selected from the group consisting of ovarian cancer, breast cancer, melanoma, colorectal cancer, gastric cancer, kidney cancer, prostate cancer, pancreatic cancer, esophageal cancer, head and neck cancer, thyroid cancer, lung cancer, bladder cancer, cervical cancer , colon cancer, liver cancer, peritoneal cancer, hepatocellular carcinoma, glioblastoma, urinary tract cancer, rectal cancer, endometrial cancer, uterine cancer, salivary gland cancer, squamous cell carcinoma, vulvar cancer, anal cancer, penile cancer , brain cancer, lymphoma (eg, B-cell lymphoma), leukemia, and metastases of the above tumors.
  • lymphoma eg, B-cell lymphoma
  • the anti-CD73 antibody or antigen-binding portion thereof of the present application can be used alone or in combination with other compositions in anti-tumor therapy.
  • an anti-CD73 antibody can be co-administered with at least one additional therapeutic agent and/or adjuvant.
  • the additional compound is a therapeutic antibody other than an anti-CD73 antibody.
  • the additional therapeutic agent is a therapeutic agent that targets the cancer immune cycle (e.g., immune checkpoint inhibitors (e.g., anti-PD-1 antibody, anti-PD-L1 antibody, or anti-CTLA4 antibody), A2AR antagonists, STAT-3 inhibitors), combination therapy with such therapeutic agents can be used to reduce tumor-mediated immunosuppression.
  • immune checkpoint inhibitors e.g., anti-PD-1 antibody, anti-PD-L1 antibody, or anti-CTLA4 antibody
  • A2AR antagonists e.g., anti-PD-1 antibody, anti-PD-L1 antibody, or anti-CTLA4 antibody
  • STAT-3 inhibitors e.g., anti-CTLA4 antibody
  • combination therapy with such therapeutic agents can be used to reduce tumor-mediated immunosuppression.
  • the present application provides a conjugate, which comprises the anti-CD73 antibody or antigen-binding portion thereof described in the first to fourth aspects and a detectable label.
  • Detectable labels are, for example, detectable fluorescent labels, chemiluminescent labels, isotopic labels, and the like.
  • the present application provides a fusion protein comprising the anti-CD73 antibody or antigen-binding portion thereof described in the first to fourth aspects.
  • antibody fusion proteins include Fab fusion proteins, Fc fusion proteins, and single-chain antibody (scFv) fusion proteins, etc., which are named according to the different sites to which effector proteins (such as cytokines) are fused.
  • a conjugate may be formed by synthesizing an anti-CD73 antibody of the present application to which a linker is covalently attached to one or more non-antibody-acting agents.
  • an antibody of the application is conjugated or recombinantly fused to a diagnostic, detection, or therapeutic agent, or any other molecule. Conjugated or recombinantly fused antibodies can be used, for example, to monitor or predict the onset, occurrence, progression and/or severity of CD73-mediated diseases as part of a clinical trial approach, eg, to determine the efficacy of a particular therapy. Such diagnosis and detection can be accomplished, for example, by conjugating the antibody to a detectable label.
  • Detectable labels include, but are not limited to, enzymes, prosthetic groups, fluorescent labels, chemiluminescent labels, isotopic labels, and the like.
  • an antibody of the present application may be conjugated or recombinantly fused to a therapeutic moiety or drug moiety that modifies a given biological response (here also an antibody drug conjugate as described in the eighth aspect).
  • Therapeutic or drug moieties are not limited to typical chemotherapeutic agents.
  • the drug moiety can be a protein, peptide, polypeptide, small molecule toxin, etc. that has the desired biological activity.
  • mice Healthy BALB/c mice were immunized with recombinant human CD73 protein (Arco, Cat. No. CD3-H52H7). After the initial immunization, booster immunization was performed three times every 14 days. The peripheral serum of the mice was collected, and the titer of the mouse serum was detected by enzyme-linked immunosorbent assay (ELISA), and mice with high plasma antibody titers were selected for cell fusion. Pulse immunizations were performed 3 days before fusion via tail vein or intraperitoneally. On the day of fusion, the mouse spleen was taken, prepared into a single cell suspension, mixed with SP2/0 cells and splenocytes at a ratio of 1:2, and fused by electrofusion.
  • ELISA enzyme-linked immunosorbent assay
  • the hybridoma cells Place them in a CO 2 incubator at 37°C and culture them. When the hybridoma cells grow to a certain amount, the hybridoma supernatants are screened by ELISA. Positive clones were selected and screened for subclones by limiting dilution. Finally, the obtained monoclonal positive cells are extracted from RNA, reverse transcription and PCR amplification to obtain the heavy chain and light chain variable region fragments of the mouse immunoglobulin for sequencing. Finally, the heavy chain and light chain CDR region sequences and the amino acid sequences of the variable regions of 7C3C2 and 1F2F9 clones were obtained as follows:
  • Heavy chain CDR1 SYDIN (SEQ ID NO.1)
  • Heavy chain CDR2 WIYPRDGFTKYNEKFKG (SEQ ID NO.2)
  • Heavy chain CDR3 KEGYRGDWYFDV (SEQ ID NO.3)
  • Light chain CDR1 RASGNVHNYLA (SEQ ID NO.5)
  • Heavy chain CDR1 SYWMH (SEQ ID NO.9)
  • Heavy chain CDR2 NINPSNGGTKYNEKFKS (SEQ ID NO.10)
  • Heavy chain CDR3 RDYGSPSY (SEQ ID NO.11)
  • Light chain CDR2 RTNILLD (SEQ ID NO.14)
  • Light chain CDR3 LQYHEFPVT (SEQ ID NO.15)
  • Embodiment 2 ELISA method detects the affinity of chimeric antibody
  • a human-mouse chimeric antibody expression vector was constructed.
  • the VH and VL were respectively constructed on eukaryotic expression vectors containing the hIgG1-kappa constant region, which contained L234A, L235A, P329A, and P331S mutations (referred to as hIgG1 in this application, and the sequence of the heavy chain constant region is shown in SEQ ID NO.39. shown, the light chain constant region sequence is shown in SEQ ID NO.40, the following examples are the same).
  • the obtained eukaryotic expression vector was transiently transformed into HEK293 cells and cultured for 6 days.
  • the culture supernatant was harvested and purified through a ProteinA column to obtain the target antibodies, named 7C3C2-hIgG1 and 1F2F9-hIgG1 respectively, and the affinity of the antibodies to recombinant human CD73 protein (same as in Example 1) was determined by ELISA.
  • the EC 50 of 7C3C2-hIgG1 and 1F2F9-hIgG1 binding to CD73 protein were 0.017nM and 0.028nM, respectively. It can be seen that the EC 50 values of these two antibodies meet the requirements of further experiments.
  • the humanized light chain template for the mouse 7C3C2 is IGKV1-NL1; for the heavy chain, the closest human matching sequence is the IGHV1-8 gene; the humanized light chain template for the mouse 1F2F9 is the IGKV1-16 gene, The closest human heavy chain match is the IGHV1-3 gene.
  • the CDRs of the light and heavy chains were respectively grafted onto the framework sequences of the matching light chain and heavy chain genes to obtain humanized antibodies Hab7C3C2 and Hab1F2F9.
  • Discovery Studio and Antibody Modeling constructs a 3D model to analyze whether there are any framework positions that replace mouse amino acids with human amino acids that affect binding and/or CDR conformation. See Table 2 and Table 3 for the reverse mutation sequence. See Table 4 and Table 5 for relevant humanized antibodies.
  • Table 4 Schematic diagram of 7C3C2 humanized antibody combination
  • Humanized antibody VH and VL were respectively constructed on eukaryotic expression vectors containing hIgG1-kappa constant region, which contained L234A, L235A, P329A, P331S mutations (abbreviated as hIgG1 in this application).
  • the affinity of the derivatized antibody was Using the HIS1K biosensor, human CD73 protein was immobilized on the biosensor to a binding response threshold of 0.3 nm. After the 120s baseline step, the sensors were respectively immersed in the anti-CD73 antibody analyte diluted with 0.02% PBST buffer, the initial antibody concentration was 25nM, 2-fold serial dilution, and 7 concentration gradients were set.
  • the binding and dissociation times of ligand and analyte were 120s and 300s, respectively. According to the binding curve, the affinity data of the antigen and antibody are calculated by regression analysis software. Among all the detected antibodies, Hab7C3C2.5, Huab7C3C.9, Huab1F2F9.12 and Huab1F2F9.15 showed the highest affinity and fewer back mutations, the results are shown in Table 6.
  • Table 6 Affinity of humanized antibody to human CD73 protein detected by Fortebio method
  • Test object EC50 value (nM) Hab7C3C2.5 0.013 Hab1F2F9.15 0.011
  • the EC 50 of Hab7C3C2.5 and Huab1F2F9.15 binding to CD73 protein were 0.013nM and 0.011nM, respectively. It can be seen that the affinity of the humanized antibody is not affected compared with the mouse antibody.
  • This example compares exemplary antibodies of the present application to anti-CD73 antibodies described in other patent applications.
  • the MEDI9447 and Hu101-28 (CDR region sequences are from PCT patent applications WO 2016/075099 and WO2018/137598, respectively) antibodies were cloned into eukaryotic expression vectors containing hIgG1-kappa constant regions.
  • the tumor cell lines MDA-MB-231 and SKOV3 endogenously expressing human CD73 on the cell surface were used to evaluate the binding ability of humanized antibodies Hab7C3C2.5 and Hab1F2F9.15 to CD73 on the cell surface.
  • the cells were seeded in a 96-well plate at a density of 2 ⁇ 10 6 /ml, 100 ⁇ l/well.
  • the cross-binding ability of the exemplary humanized anti-CD73 antibody of the present application and cynomolgus monkey CD73 was detected by Fortebio assay.
  • the specific implementation steps include immobilizing the CD73 protein of cynomolgus monkey on the HIS1K biosensor. After the 120s baseline step, the sensors were respectively immersed in the anti-CD73 antibody analyte diluted with 0.02% PBST buffer. The initial concentration of the antibody was 25nM, and 7 concentration gradients were set respectively for affinity analysis experiments. The binding and dissociation times of ligand and analyte were 120s and 300s, respectively. According to the binding curve, the affinity data of the antigen and antibody were calculated by regression analysis software. The results are shown in Table 9, the exemplary antibody of the present application can cross-recognize the CD73 protein of cynomolgus monkey.
  • Table 9 Affinity of humanized antibody to cynomolgus monkey CD73 protein detected by Fortebio method
  • CHO-K1 cells that can stably express the monkey-derived full-length CD73 were constructed.
  • the cells were seeded in a 96-well plate at a density of 2 ⁇ 10 6 /ml, 100 ⁇ l/well. Wash three times with PBS buffer, discard the supernatant.
  • Antibody diluted in PBS buffer was added and incubated at 4°C for 1 h.
  • the cells were washed 3 times with PBS buffer, and then labeled with Alexa Fluor 488-labeled goat anti-human IgG antibody at 4°C in the dark for 50 min.
  • the cells were washed 3 times with PBS buffer, and the fluorescence intensity of the cells was read by flow cytometry.
  • the prepared humanized antibody can dose-dependently bind to the monkey expressed on the cell CD73 protein.
  • Table 10 The exemplary humanized antibody of the present application and the binding experiment of cynomolgus monkey CD73 on the cell surface
  • CD73-expressing MDA-MB-231 cells were diluted at 2 ⁇ 10 5 /ml, 100 ⁇ l/well was inoculated into a 96-well plate. The plate was incubated at 37° C. for 1 h, then AMP (1800 ⁇ M) was added to each well, 50 ⁇ l/well, and incubated at 37° C. for 2 hours.
  • the cells were centrifuged at 1000 rpm for 5 min, and 50 ⁇ l of the supernatant was transferred to a new 96-well plate, and 50 ⁇ l of ATP was added to make the final concentration 75 ⁇ M.
  • CellTiter-Glo reagent Promega was added at a ratio of 1:1 to measure the cellular CD73 enzyme activity. The results are shown in Figure 7 and Table 12: Hab7C3C.5 and Hab1F2F9.15 can inhibit the CD73 enzyme activity on MDA-MB-231 cells.
  • antibody-mediated internalization of CD73 was assessed by flow cytometry.
  • the cells were suspended in serum-free DMEM medium, and incubated with the tested antibodies for 1 h on ice at a concentration of 10 ⁇ g/ml.
  • the cells were washed three times with serum-free DMEM medium; the cells incubated with different antibodies were suspended in serum-free DMEM medium, and each was divided into two parts and spread in two 24-well plates.
  • One culture plate was incubated in a 37°C incubator; the other culture plate was placed in a 4°C refrigerator as a control group; cultured for 24 hours. Collect and wash the cells, add Alexa Fluor 488-labeled goat anti-human secondary antibody, and stain at 4°C for 1 h in the dark. Washed 3 times in PBS buffer and detected by flow cytometry.
  • the exemplary antibody of the present application can induce about 60% of CD73 receptor internalization, and maintain about 40% of the initial surface staining. All calculated values are percent internalization compared to the control (set as 100%).
  • the C-terminal part (SEQ ID NO.41) and the N-terminal part (SEQ ID NO.42) of the CD73 protein were respectively constructed, and the antibody recognition epitope was detected by ELISA method.
  • the CD73 N-terminal and C-terminal proteins were coated on the microplate at a concentration of 1 ⁇ g/ml, and the antibodies to be tested were serially diluted. The resulting affinity is shown in Figure 9A (N-terminal) and 9B (C-terminal).
  • the experimental results show that: compared with the control antibodies (MEDI9447 and Hu101-28), the binding epitopes of Hab7C3C.5 and Hab1F2F9.15 are different; Bind to the N-terminus of CD73 protein, and Hu101-28 binds to the C-terminus of CD73 protein.
  • the binding epitopes of Hab7C3C.5 and Hab1F2F9.15 of the present application are obviously different from those of MEDI9447 and Hu101-28.
  • the monotherapy efficacy of the 1F2F9 chimeric antibody was evaluated in a tumor xenograft model.
  • 4 ⁇ 106 A375 melanoma cells and 4 ⁇ 105 human PBMCs were mixed with Matrigel 1:1 and inoculated into NCG immunodeficient mice.
  • 30 mg/kg of 1F2F9 chimeric antibody or PBS control was injected intraperitoneally and every other day.
  • 24 days after treatment several mice had tumors that reached an upper limit (2000 mm 3 ). Day 24 after treatment therefore became the data endpoint for the analysis.
  • the mean tumor volume was 1448 ⁇ 176.76 mm 3 (mean ⁇ standard error) at 24 days of treatment.
  • human CD73/PD1/PDL1 knock-in BALB/c mice and CT26 cells (colon cancer cell line) overexpressing human CD73/PDL1 were used to evaluate Huab1F2F9.15, anti-PD-1 antibody and their combination efficacy.
  • 1 ⁇ 10 6 CT26-hCD73/hPDL1 cells were subcutaneously inoculated into genetic mice.
  • the day of grouping was defined as day D0, and administration began on the day of grouping.
  • the indicated doses of antibodies or PBS control were injected intraperitoneally twice a week.
  • the present application has obtained an antibody with high affinity that recognizes CD73 protein and CD73 at the cellular level.
  • the representative antibodies tested can potently inhibit the enzymatic activity of CD73, and at the same time induce the internalization of CD73 protein on tumor cells, and further reduce the enzymatic activity of CD73 by reducing the CD73 molecules on the cell surface, thereby achieving inhibition through multiple pathways Effect of CD73 enzymatic activity.
  • Epitope analysis shows that the exemplary antibody of the present application does not bind to either the N-terminal or the C-terminal protein of the CD73 protein alone, and has differential epitope binding characteristics from prior art antibodies such as MEDI9447 and Hu101-28.

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Abstract

本申请涉及新的抗CD73抗体及其用途，所述抗CD73抗体可用于***，特别是肿瘤细胞表面表达CD73(CD73+)的肿瘤。

Description

抗CD73抗体及应用
相关申请的引用
本申请要求于2022年1月25日递交的中国专利申请第202210098976.1号的优先权，通过援引加入的方式将其全部内容整体并入本文，并用于所有目的。
技术领域
本申请大体涉及生物制药领域，具体而言，本申请涉及新的抗CD73抗体或其抗原结合片段、包含所述抗体或其抗原结合片段的药物组合物及所述抗体或其抗原结合片段的医药学用途。
背景技术
CD73是由NT5E基因编码的胞外-5’-核苷酸酶，包含N端及C端两部分，通过糖基磷脂酰肌醇(GPI)锚定于细胞膜外表面形成同源二聚体。CD73广泛表达于人体内皮细胞，淋巴细胞等细胞表面。在胃癌、肾癌、***癌、乳腺癌(例如三阴性乳腺癌)、非小细胞腺癌等多种肿瘤组织中呈现高表达，且提示预后不良。CD73可将细胞外腺苷单磷酸(AMP)水解为腺苷。腺苷是一种功能强大的免疫抑制性分子，通过抑制保护性免疫细胞(如效应性T细胞、NK细胞、DC和B细胞)的功能，同时维持调节性免疫细胞(如Treg、MDSC、TAM和CAF)的功能来削弱抗肿瘤免疫，参与肿瘤的免疫逃逸。除了抑制免疫细胞功能，越来越多的研究表明CD73也可以直接刺激肿瘤细胞增殖、迁移、侵袭及肿瘤血管生成，抑制胞外核苷酸酶CD73可以逆转腺苷介导的免疫抑制。研究发现，低氧及放化疗导致PDL1、CD73和CD47表达上调。因此，靶向CD73的治疗策略有潜力作为单药或联合疗法应用于临床。目前有近30种抗CD73抗体产品在研，包括Medimmune的oleclumab和天境生物的uliledlimab。进展最快的CD73抗体处于临床三期，无上市产品，因此，针对CD73抗体的开发和应用是本领域亟需的。
发明概述
第一方面，本申请提供了抗CD73抗体或其抗原结合部分，所述抗CD73抗体或其抗原结合部分包括重链可变区和轻链可变区，所述重链可变区包括重链CDR1(HCDR1)、重链CDR2(HCDR2)和重链CDR3(HCDR3)，轻链可变区包括轻链CDR1(LCDR1)、轻链CDR2(LCDR2)和轻链CDR3(LCDR3)，其中：
HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.2所示序列或与SEQ ID NO.2所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.6所示序列或与SEQ ID NO.6所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.17所示序列或与SEQ ID NO.17所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.32所示序列或与SEQ ID NO.32所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID  NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.17所示序列或与SEQ ID NO.17所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.6所示序列或与SEQ ID NO.6所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.2所示序列或与SEQ ID NO.2所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.32所示序列或与SEQ ID NO.32所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.21所示序列或与SEQ ID NO.21所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.32所示序列或与SEQ ID NO.32所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.21所示序列或与SEQ ID NO.21所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.6所示序列或与SEQ ID NO.6所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.23所示序列或与SEQ ID NO.23所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.32所示序列或与SEQ ID NO.32所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.23所示序列或与SEQ ID  NO.23所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.6所示序列或与SEQ ID NO.6所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.10所示序列或与SEQ ID NO.10所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.13所示序列或与SEQ ID NO.13所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.10所示序列或与SEQ ID NO.10所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.35所示序列或与SEQ ID NO.35所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.27所示序列或与SEQ ID NO.27所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.35所示序列或与SEQ ID NO.35所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.27所示序列或与SEQ ID NO.27所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.13所示序列或与SEQ ID NO.13所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.30所示序列或与SEQ ID NO.30所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同 一性，LCDR1的序列为SEQ ID NO.35所示序列或与SEQ ID NO.35所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.30所示序列或与SEQ ID NO.30所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.13所示序列或与SEQ ID NO.13所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；
其中HCDR和LCDR序列根据Kabat定义。
第二方面，本申请提供了抗CD73抗体或其抗原结合部分，所述抗CD73抗体或其抗原结合部分包括重链可变区和轻链可变区，其中：
所述重链可变区具有SEQ ID NO.4、18、19、20、22、24、12、25、26、28、29或31所示氨基酸序列或与SEQ ID NO.4、18、19、20、22、24、12、25、26、28、29或31所示氨基酸序列具有至少80％、至少85％、至少90％、至少95％或至少99％同一性的氨基酸序列；和/或
所述轻链可变区具有SEQ ID NO.8、33、34、16、36、37或38所示氨基酸序列或与SEQ ID NO.8、33、34、16、36、37或38所示氨基酸序列具有至少80％、至少85％、至少90％、至少95％或至少99％同一性的氨基酸序列。
第三方面，本申请提供了抗CD73抗体或其抗原结合部分，所述抗CD73抗体或其抗原结合部分包括重链可变区，所述重链可变区包括重链CDR1(HCDR1)、重链CDR2(HCDR2)和重链CDR3(HCDR3)，其中：
(1)HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.2所示序列或与SEQ ID NO.2所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性；或
(2)HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.17所示序列或与SEQ ID NO.17所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性；或
(3)HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.21所示序列或与SEQ ID NO.21所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性；或
(4)HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.23所示序列或与SEQ ID NO.23所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同 一性；或
(5)HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.10所示序列或与SEQ ID NO.10所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性；或
(6)HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.27所示序列或与SEQ ID NO.27所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性；或
(7)HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.30所示序列或与SEQ ID NO.30所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性；
其中HCDR序列根据Kabat定义。
第四方面，本申请提供了抗CD73抗体或其抗原结合部分，所述抗CD73抗体或其抗原结合部分包括轻链可变区，轻链可变区包括轻链CDR1(LCDR1)、轻链CDR2(LCDR2)和轻链CDR3(LCDR3)，其中：
(1)LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.6所示序列或与SEQ ID NO.6所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
(2)LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.32所示序列或与SEQ ID NO.32所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
(3)LCDR1的序列为SEQ ID NO.13所示序列或与SEQ ID NO.13所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；或
(4)LCDR1的序列为SEQ ID NO.35所示序列或与SEQ ID NO.35所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；
其中LCDR序列根据Kabat定义。
第五方面，本申请提供了分离的核酸分子，其编码第一方面至第四方面所述的抗CD73抗体或其抗原结合部分。
第六方面，本申请提供了载体，其含有第五方面所述的核酸分子。
第七方面，本申请提供了宿主细胞，其含有第五方面所述的核酸分子或第六方面所述的载体。
第八方面，本申请提供了抗体药物偶联物，其包含与治疗剂偶联的第一方面至第四方面所述的抗CD73抗体或其抗原结合部分。
第九方面，本申请提供了药物组合物，其包含第一方面至第四方面所述的抗CD73抗体或其抗原结合部分或第八方面所述的抗体药物偶联物，以及药学可接受的载体。
第十方面，本申请提供了第一方面至第四方面所述的抗CD73抗体或其抗原结合部分、第五方面所述的核酸分子、第六方面所述的载体、第七方面所述的宿主细胞或第八方面所述的抗体药物偶联物在制备用于治疗个体中的肿瘤，特别是肿瘤细胞表面表达CD73(CD73+)的肿瘤的药物中的用途。
第十一方面，本申请提供了治疗个体中的肿瘤，特别是肿瘤细胞表面表达CD73(CD73+)的肿瘤的方法，所述方法包括给予所述个体治疗有效量的第一方面至第四方面所述的抗CD73抗体或其抗原结合部分、第八方面所述的抗体药物偶联物或第九方面所述的药物组合物。
第十二方面，本申请提供了缀合物，其包含第一方面至第四方面所述的抗CD73抗体或其抗原结合部分以及可检测标记物。
第十三方面，本申请提供了包含第一方面至第四方面所述的抗CD73抗体或其抗原结合部分的融合蛋白。
附图简要描述
图1显示了本申请的示例性抗体与可溶性CD73的结合能力测试结果。
图2显示了本申请的示例性人源化抗体与可溶性CD73的结合能力测试结果。
图3显示了本申请的示例性人源化抗体与人乳腺癌MDA-MB-231细胞表面上CD73的结合能力测试结果。
图4显示了本申请的示例性人源化抗体与人卵巢癌SKOV3细胞表面上CD73的结合能力测试结果。
图5显示了本申请的示例性人源化抗体与细胞表面食蟹猴CD73的结合能力测试结果。
图6显示了本申请的示例性人源化抗体对CD73蛋白的酶活抑制实验结果。
图7显示了本申请的示例性人源化抗体对细胞表面CD73蛋白的酶活抑制实验结果。
图8显示了本申请的示例性人源化抗体诱导CD73内化的实验结果。
图9A显示了本申请的示例性人源化抗体识别CD73蛋白N端的检测结果。
图9B显示了本申请的示例性人源化抗体识别CD73蛋白C端的检测结果。
图10A显示了本申请的示例性抗体以及其他测试物的给药产生的A375荷瘤小鼠中移植瘤的体积变化结果。
图10B显示了本申请的示例性抗体以及其他测试物的给药产生的个体A375荷瘤小鼠中移植瘤体积的变化结果。
图10C显示了本申请的示例性抗体以及其他测试物的给药对A375荷瘤小鼠体重的影响结果。
图11A显示了本申请的示例性人源化抗体以及其他测试物的给药产生的CT26荷瘤小鼠中移植瘤的体积变化结果。
图11B显示了本申请的示例性人源化抗体以及其他测试物的给药产生的个体CT26荷瘤小鼠中移植瘤体积的变化结果。
图11C显示了本申请的示例性人源化抗体以及其他测试物给药对CT26荷瘤小鼠体重的影响结果。
发明详细描述
定义
除非另有定义，本文使用的所有科技术语具有本领域普通技术人员所理解的相同含义。关于本领域的定义及术语，专业人员具体可参考Current Protocols in Molecular Biology(Ausubel)。氨基酸残基的缩写是本领域中所用的指代20个常用L-氨基酸之一的标准3字母和/或1字母代码。
尽管本申请的广义范围所示的数字范围和参数近似值，但是具体实施例中所示的数值尽可能准确的进行记载。然而，任何数值本来就必然含有一定的误差，其是由它们各自的测量中存在的标准偏差所致。另外，本文公开的所有范围应理解为涵盖其中包含的任何和所有子范围。例如记载的“1至10”的范围应认为包含最小值1和最大值10之间(包含端点)的任何和所有子范围；也就是说，所有以最小值1或更大起始的子范围，例如1至6.1，以及以最大值10或更小终止的子范围，例如5.5至10。另外，任何称为“并入本文”的参考文献应理解为以其整体并入。
本文所使用的术语“对象”或“个体”是指哺乳动物，如人类，但也可以是其它动物，如野生动物，家畜或实验动物(如猩猩、猴子、大鼠、小鼠、兔子、豚鼠、土拨鼠、地松鼠等)。
本文所使用的术语“抗原”是抗体可选择性与之结合的预定的靶标。抗原的实例包括但不限于多肽、糖、核酸、脂质、半抗原或其它天然存在的化合物或合成化合物。
广义而言，“抗体”可以指能够经由至少一个位于免疫球蛋白分子的可变区中的抗原识别位点特异性结合到靶标的免疫球蛋白分子，因此涵盖完整抗体/全长抗体、抗体单链或抗体的任何抗原结合片段(也称为“抗原结合部分”)。当“抗体”和“抗原结合片段/抗原结合部分”出现在同一语境下时，“抗体”可以理解为相对于“抗原结合片段/抗原结合部分”的完整体，二者共同对应于广义的抗体概念。
术语“抗CD73抗体”或“与CD73结合的抗体”包括能够以足够的亲和力结合CD73，使得抗体在靶向CD73时可用作诊断剂和/或治疗剂的抗体。在一些实施方案中，抗CD73抗体与无关的非CD73蛋白结合的程度小于抗体与CD73结合的约10％，例如通过荧光激活细胞分选(FACS)分析或免疫测定法(例如放射免疫测定法(RIA))测定。对于与CD73“特异性结合”或对CD73“具有特异性”的抗体，在某些实施方案中，与CD73结合的抗体的解离常数(KD)小于或等于500nM、100nM、10nM、9nM、8nM、7nM、6nM、5nM、4nM、0.9nM、0.8nM、0.7nM、0.6nM、0.5nM、0.4nM、0.3nM、0.2nM或0.1nM。在一些实施方案中，抗CD73抗体与来自不同物种的CD73间(例如人和食蟹猴CD73间)保守的CD73蛋白表位结合。
术语“激动剂抗体”是引发反应的抗体，例如，模拟目标多肽的至少一种功能活性的抗体。激动剂抗体包括是配体模拟物的抗体，例如，其中配体与细胞表面受体结合，所述结合通过胞内细胞信号转导途径诱导细胞信号转导或活性，且其中抗体诱导类似的细胞信号转导或活化。
“全长抗体”系指包含通过二硫键而互连的至少两条重(H)链和两条轻(L)链的蛋白。每条重链包含一重链可变区(缩写为VH)和一重链恒定区。该重链恒定区包含三个域(domain)，CH1、CH2和CH3。每条轻链包含一轻链可变区(缩写为VL)和一轻链恒定区。该轻链恒定区包含一个域，CL。VH和VL区域还可再细分为具有高可变性的多个区，被称为互补决定区(CDR)，其间散布有更为保守的被称为框架区(FR)的多个区域。每个VH和VL均由三个CDR和四个FR构成，按照以下顺序从氨基端至羧基端排布：FR1、CDR1、FR2、CDR2、FR3、CDR3、FR4。重链和轻链的这些可变区包含与抗原相互作用的结合域。抗体的恒定区可介导免疫球蛋白与宿主的组织或因子结合，包括免疫***的各种细胞(如效应细胞)和经典补体***的第一成分(Clq)。全长抗体可以是任何种类的抗体，例如IgD、IgE、IgG、IgA或IgM(或上述的子类)，但抗体不需要属于任何特定的类别。根据重链的恒定域的抗体氨基酸序列，可以将免疫球蛋白指定为不同的类别。通常，免疫球蛋白有五种主要的类别：IgA、IgD、IgE、IgG及IgM，而且这些类别中有几个可以再被进一步区分成子类(同型)，例如IgG1、IgG2、IgG3、IgG4、IgA1及IgA2。对应于不同免疫球蛋白类别的重链恒定域分别称为α、 δ、ε、γ、以及μ。不同类别的免疫球蛋白的子单元结构和三维结构是公知的。嵌合或人源化抗体也涵盖在根据本申请的抗体中。本领域技术人员公知，互补决定区(CDR，通常有CDR1、CDR2及CDR3)是可变区中对抗体的亲和力和特异性影响最大的区域。VH或VL的CDR氨基酸序列有多种常见的定义方式，例如Kabat定义、IMGT定义，Chothia定义等。对于给定抗体的可变区氨基酸序列，通常可以根据不同定义方式来确定VH和VL氨基酸序列中的CDR氨基酸序列。在本申请的实施方案中，利用Kabat义CDR氨基酸序列。对于给定抗体的可变区氨基酸序列，可以通过多种方式分析可变区氨基酸序列的中CDR氨基酸序列。
术语“鼠源抗体”是指一种抗体，其来源于免疫接种过的小鼠的B细胞与骨髓瘤细胞融合，筛选出既能无限增殖又能分泌抗体的鼠杂交融合细胞，进而进行筛选、制备和纯化获得的抗体，鼠源抗体一般具有免疫原性，因此后续需要对其作人源化处理。
术语“人源化抗体”意指将源自另一哺乳动物物种，诸如小鼠种系的CDR序列嫁接到人框架序列上获得的抗体。为了保留结合亲和力，可以修饰骨架(称为FR)区段的一些残基。通过本领域技术人员已知的技术可以制备根据本申请的人源化抗体或其片段；
术语“嵌合抗体”系指以下抗体，其中的可变区序列来自一物种而恒定区序列来自另一物种，例如可变区序列来自小鼠抗体而恒定区序列来自人抗体的抗体。通过使用基因重组技术可以制备根据本申请的嵌合抗体或其片段。例如，所述嵌合抗体可以通过克隆重组DNA来生产，所述重组DNA包含启动子和编码根据本申请的非人尤其是鼠单克隆抗体可变区的序列、以及编码人抗体恒定区的序列。由这种重组基因编码的本申请嵌合抗体将是，例如，鼠-人嵌合体，该抗体的特异性由来源于鼠DNA的可变区确定，并且其同种型由来源于人DNA的恒定区来确定。
术语“部分人源化抗体”指含有来自人的恒定区和来自非人(如鼠)的可变区(包括CDR)的抗体。
术语“半人源化抗体”是人源化抗体的一种，是指其中一个抗体链包含鼠可变区，并且另一抗体链包含人源化可变区的抗体，即半人源化抗体。
术语“单克隆抗体”指由基本同质的抗体群体获得的抗体，即，除了可能在少量个体中存在自然发生的突变以外，组成群体的各个抗体是相同的。
本文所使用的术语“抗原结合片段”或“抗原结合部分”或“抗原结合区”可互换使用，是指包含与抗原相互作用并赋予结合剂其针对抗原的特异性和亲和力的氨基酸残基的抗体的部分，尤其是指抗体片段如Fv、Fab、F(ab’)2或Fab’，或者通过化学修饰或通过掺入脂质体中应能够增加半寿期的任何片段，所述化学修饰例如添加聚(亚烷基)二醇如聚乙二醇(“聚乙二醇化，PEG化”)(被称为Fv-PEG、scFv-PEG、Fab-PEG、F(ab’)2-PEG或Fab’-PEG的聚乙二醇化片段)(“PEG”为聚乙二醇)，所述片段具有CD73结合活性。优选地，所述功能片段将由其来源抗体的重链或轻链可变链的部分序列构成或者包含它们，所述部分序列足以保留与其来源抗体相同的结合特异性和充分的亲和力，这种功能片段将包含最少5个氨基酸，优选其来源的抗体序列的10、15、25、50和100个连续氨基酸。抗原结合片段的实例包括但不限于：(1)Fab片段，其可以是具有VL-CL链和VH-CH1链的单价片段；(2)F(ab')2片段，其可以是具有两个Fab'片段的二价片段，该两个Fab'片段由铰链区的二硫桥(即Fab'的二聚物)连接；(3)具有抗体的单臂的VL和VH域的Fv片段。
术语“单链抗体(scFv)指是由VH域和VL域经由肽连接子连接的单一多肽链。(scFv)2包含两个由肽连接子连接的VH域和两个VL域，该两个VL域是经由二硫桥与该两个VH域组合。
术语“Fc片段”、“Fc区”、“Fc结构域”、“Fc部分”或类似的术语是指抗体重链恒定区的一部分，包括铰链区(hinge)、恒定区的CH2片段和CH3片段。可以对抗CD73抗体的Fc区进行工程化改造或修饰，包括效应功能相关的改造，以例如降低或消除抗体依赖的细胞 毒性(ADCC)和/或依赖补体的细胞毒性(CDC)，这可通过在抗体的Fc区引入一个或更多个氨基酸取代/突变来实现。
如本文所用术语“特异性结合”是指两个分子之间的非随机结合反应，例如抗体与抗原表位的结合。
术语“多抗”又称“多特异性抗体”，是具有针对至少2种不同抗原的结合特异性的分子，其中，仅结合2种抗原的该类分子也被称为双抗(即双特异性抗体，BsAb)。
术语“双特异性抗体”是指同时具有两种抗原表位结合能力的抗体。两种抗原表位可以在不同抗原上，也可是在同一抗原上。双特异性抗体可以具有多种结构构型。例如，双特异性抗体可以由两个Fc片段以及分别与其融合的两个抗原结合部分组成(与天然抗体相似，区别在于两个臂结合不同的抗原靶标或表位)，抗原结合部分可以为单链抗体(scfv)的形式或者Fab片段的形式。
通常，为了制备单克隆抗体或其功能片段，尤其是鼠源的单克隆抗体或其功能片段，可以参考尤其描述在手册“Antibodies”中的技术(Harlow and Lane，Antibodies：A Laboratory Manual，Cold Spring Harbor Laboratory，Cold Spring Harbor NY，pp.726，1988)或者参考Kohler和Milstein描述的从杂交瘤细胞制备的技术(Nature，256：495-497，1975)。
术语“保守变体”或“保守氨基酸置换”是基本上不影响或降低蛋白的亲和力例如抗体对CD73的亲和力的那些替换。例如，特异性结合CD73的人抗体可包括至多约1、至多约2、至多约5、至多约10或至多约15个保守替换，并且特异性结合CD73多肽。术语“保守变体”还包括使用取代的氨基酸代替未取代的亲本氨基酸，只要抗体特异性结合CD73。非保守替换是降低活性或与CD73结合的那些。
在描述涉及氨基酸或核酸序列突变的一些实施方案时，本申请使用“XaaaY”的限定方式(例如L234A、L235A等)，其中“aaa”代表氨基酸或碱基的序列位置(在存在具体参考序列时，“aaa”代表在该参照序列中的残基顺序位置；或者也可以按照本领域通行的位置编号方式，例如EU编号***等)，“X”代表“aaa”处原有的氨基酸或碱基，“Y”代表“aaa”处变化后的氨基酸或碱基。作为实例，当描述人重链恒定区的L234A突变时，是指按照人重链恒定区的EU编号***，第234位的亮氨酸(L)被突变为丙氨酸(A)。
本文中，当描述药物A和B进行的“联合治疗”、“组合治疗”或类似术语时，除非在技术上冲突或不兼容，药物A和B联合治疗的给药方式和方案不受特定的限制。例如，药物A和B可以依次分别给药、同时分别给药、配制于同一药剂中一并给药。或者，药物A和B可以在生产时即制备于同一组合物中，或同一药品套装中。药物A和B在物理上可以是分隔的，或者结合在一起。药物A和B的给药在时间上可以是分开的、同时的或重叠的。
术语“分离的”生物组分(例如核酸、蛋白质(包括抗体)或细胞器)已经与所述组分天然存在的环境(例如细胞)中的其他生物组分(即，其他染色体和额外染色体DNA和RNA、蛋白质和细胞器)基本分离或被纯化。已经被“分离”的核酸和蛋白质包括用标准纯化方法纯化的核酸和蛋白质。该术语还包括通过在宿主细胞中重组表达制备的核酸和蛋白质以及化学合成的核酸。
术语“衍生序列”是指与相关序列具有至少80％(优选85％、90％、95％、98％或者99％)序列同一性且仍具有相同或相似功能的序列。
本文使用的术语“药物组合物”其表示组合在一起以实现某种特定目的的至少一种药物以及任选地可药用载体或辅料的组合。在某些实施方案中，所述药物组合物包括在时间和/或空间上分开的组合，只要其能够共同作用以实现本申请的目的。例如，所述药物组合物中所含的成分(例如根据本申请的抗体、核酸分子、核酸分子组合和/或缀合物)可以以整体施用于个体，或者分开施用于个体。当所述药物组合物中所含的成分分开地施用于个体时，所述成分可以同时或依次施用于个体。优选地，所述可药用载体是水、缓冲水溶液、等渗 盐溶液如PBS(磷酸盐缓冲液)、葡萄糖、甘露醇、右旋葡萄糖、乳糖、淀粉、硬脂酸镁、纤维素、碳酸镁、0.3％甘油、透明质酸、乙醇或聚亚烷基二醇如聚丙二醇、甘油三酯等。所用可药用载体的类型尤其依赖于根据本申请的组合物是否配制为用于口服、鼻、皮内、皮下、肌内或静脉施用。根据本申请的组合物可包含润湿剂、乳化剂或缓冲液物质作为添加剂。根据本申请的药物组合物或者药物制剂可通过任何适宜的途径施用，例如可口服、鼻、皮内、皮下、肌内或静脉内施用。
本文使用的术语“治疗有效量”或“有效量”是指足以显示其对于所施用个体益处的剂量。施用的实际量，以及施用的速率和时间过程会取决于所治疗者的自身情况和严重程度。治疗的处方(例如对剂量的决定等)最终是全科医生及其它医生的责任并依赖其做决定，通常考虑所治疗的疾病、患者个体的情况、递送部位、施用方法以及对于医生来说已知的其它因素。
EC50值主要是指：指在特定暴露时间后，能达到50％最大生物效应对应的药物、抗体或者毒素等的浓度。药学中除了用于表征体外实验中(in vitro)激动剂(agonist)的激活能力外，还可用于表示达到体内(in vivo)最大生物效应一半时所需的血药浓度。在一些文献中，EC50也用于表征某化合物在细胞水平的效力(包括激动和拮抗)，可以通过ELISA等方法测定EC50值。
本文使用的术语“融合蛋白”在一般语境中是由至少两个结构域组成的蛋白，各结构域不是天然状态下相关联的，由单独的基因编码，各个基因相连接，作为一个整体被转录和翻译，产生单个蛋白。在本申请的技术背景下，包含抗体或抗原结合部分的“融合蛋白”指利用基因工程技术将抗体或抗原结合部分与另外的生物活性蛋白融合所得的产物，这类抗体融合蛋白兼具抗体的抗原结合能力以及与抗体融合的生物活性蛋白的独特生物学特性。
关于氨基酸或核酸序列的术语“同一性/同源性/一致性”被定义为经过序列比对和引入空位后，氨基酸或核苷酸序列变体中相同的残基的百分比，如果需要，达到最大百分比的同一性。用于比对的方法和计算机程序在本领域内是公知的。
本文所用术语“肿瘤”是指由异常细胞生长形成的赘生物或实体病变。肿瘤可以是良性的、恶变前的或恶性的。
本文所用术语“恶性肿瘤”是指或描述哺乳动物的生理条件，其典型的特征在于不受调控的细胞生长。示例性恶性肿瘤包括：癌、实体瘤、黑素瘤肉瘤、血液肿瘤、生殖细胞瘤和胚细胞瘤。恶性肿瘤的更多具体实例包括：多发性骨髓瘤、肾癌，包括小细胞肺癌、非小细胞肺癌、肺腺癌和肺鳞癌的肺癌，膀胱癌，乳腺癌，***，结肠癌，包括胃肠癌的胃癌，***癌，胰腺癌，腹膜癌，肝细胞癌，成胶质细胞瘤，卵巢癌，肝癌，尿路癌，肝细胞瘤，直肠癌，结肠直肠癌，子宫内膜或子宫癌，唾液腺癌，鳞状细胞癌(例如，鳞状上皮细胞癌)，外阴癌，甲状腺癌，***癌，***癌，黑素瘤和B细胞淋巴瘤，脑癌及头颈癌以及相关转移灶。
本文所用术语“血液肿瘤”是指由于异常细胞的生长增殖不受控制引起的，在多数情况下这些不正常的细胞的起源部位是骨髓，这也正是血液细胞产生的地方。示例性血液肿瘤包括各类白血病、多发性骨髓瘤以及恶性淋巴瘤。血液肿瘤的更多具体实例包括：急性成淋巴细胞性白血病(ALL)、慢性淋巴细胞性白血病(CLL)、急性髓细胞性白血病(AML)、慢性髓细胞性白血病(CML)、毛细胞性白血病(HCL)、T细胞幼淋巴细胞性白血病、大颗粒淋巴细胞白血病、幼年型粒-单核细胞白血病、B细胞幼淋巴细胞性白血病、伯基特白血病和成人T细胞性白血病、非霍奇金淋巴瘤、B细胞淋巴瘤、小淋巴细胞性淋巴瘤、淋巴浆细胞淋巴瘤、原发性巨球蛋白血症(macroglobulinemia)、脾边缘区淋巴瘤、浆细胞瘤、结外边缘区B细胞淋巴瘤、MALT淋巴瘤、结内边缘区B细胞淋巴瘤(NMZL)、滤泡性淋巴瘤、套细胞淋巴瘤、弥漫性大B细胞淋巴瘤、纵隔(胸腺)大B细胞淋巴瘤、血管内大B细胞淋巴瘤、原发性渗出性淋巴瘤、伯基特淋巴瘤、B细胞慢性淋巴细胞性淋巴瘤、经典霍奇金淋巴瘤、结节性淋巴细胞为主型霍奇金淋巴瘤、成人T细胞淋巴瘤、结外鼻型 NK/T细胞淋巴瘤、肠病型T细胞淋巴瘤、肝脾T细胞淋巴瘤、母细胞NK细胞淋巴瘤、蕈样真菌病、赛谢综合征、原发性皮肤CD30阳性T细胞淋巴增生病、原发性皮肤间变性大细胞淋巴瘤、淋巴瘤样丘疹病、血管免疫母细胞性T细胞淋巴瘤、非特指型外周T细胞淋巴瘤和间变性大细胞淋巴瘤。
本文所用术语“实体瘤”是指可通过临床检查如X线摄片、CT扫描、B超或触诊扪及到的有形肿块。临床上诊治过的实体瘤分恶性和良性两种。恶性实体瘤包括：儿童霍奇金淋巴瘤：淋巴细胞为主型、结节硬化型、混合细胞型、淋巴细胞消减型；儿童非霍奇金淋巴瘤：前淋巴母细胞淋巴瘤、小无裂细胞淋巴瘤(伯基特/非伯基特淋巴瘤)、弥漫性大B细胞淋巴瘤、间变性大细胞淋巴瘤等；儿童肾脏肿瘤：肾母细胞瘤(Wilms瘤)、肾透明细胞癌、肾横纹肌样瘤、肾透明细胞肉瘤、肾原始神经外胚叶瘤等；儿童神经母细胞瘤：神经母细胞瘤、节细胞神经母细胞瘤、节细胞神经瘤；儿童颅外生殖细胞瘤：成熟畸胎瘤、未成熟畸胎瘤、内胚窦瘤(卵黄囊瘤)、***瘤、无性细胞瘤、绒毛膜上皮癌、胚胎癌等；骨肉瘤及软骨肉瘤；儿童横纹肌肉瘤：胚胎型、腺泡型、多形型等；儿童软组织肉瘤：纤维肉瘤、恶性纤维组织细胞瘤、脂肪肉瘤、平滑肌肉瘤、血管肉瘤、***肉瘤、恶性神经鞘瘤、腺泡状软组织肉瘤、上皮样肉瘤、透明细胞肉瘤、恶性黑色素瘤、滑膜肉瘤、促纤维增生性小圆细胞瘤等；尤文氏家族肉瘤：尤文氏肉瘤、原始神经外胚叶瘤；儿童肝脏肿瘤：肝母细胞瘤(胚胎型、胎儿型、未分化型)、肝细胞癌；视网膜母细胞瘤；其他肿瘤：后颅窝髓母细胞瘤、鼻咽癌、甲状腺***状癌、胸腺瘤、肺母细胞瘤、胰母细胞瘤、胰岛细胞瘤、回盲部类癌、间皮瘤等。良性实体瘤包括：***瘤、血管瘤、甲状舌管囊肿等。
第一方面，本申请提供了抗CD73抗体或其抗原结合部分，所述抗CD73抗体或其抗原结合部分包括重链可变区和轻链可变区，所述重链可变区包括重链CDR1(HCDR1)、重链CDR2(HCDR2)和重链CDR3(HCDR3)，轻链可变区包括轻链CDR1(LCDR1)、轻链CDR2(LCDR2)和轻链CDR3(LCDR3)，其中：
HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.2所示序列或与SEQ ID NO.2所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.6所示序列或与SEQ ID NO.6所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.17所示序列或与SEQ ID NO.17所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.32所示序列或与SEQ ID NO.32所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.17所示序列或与SEQ ID NO.17所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.6所示序列或与SEQ ID NO.6 所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.2所示序列或与SEQ ID NO.2所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.32所示序列或与SEQ ID NO.32所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.21所示序列或与SEQ ID NO.21所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.32所示序列或与SEQ ID NO.32所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.21所示序列或与SEQ ID NO.21所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.6所示序列或与SEQ ID NO.6所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.23所示序列或与SEQ ID NO.23所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.32所示序列或与SEQ ID NO.32所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.23所示序列或与SEQ ID NO.23所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.6所示序列或与SEQ ID NO.6所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.10所示序列或与SEQ ID  NO.10所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.13所示序列或与SEQ ID NO.13所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.10所示序列或与SEQ ID NO.10所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.35所示序列或与SEQ ID NO.35所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.27所示序列或与SEQ ID NO.27所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.35所示序列或与SEQ ID NO.35所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.27所示序列或与SEQ ID NO.27所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.13所示序列或与SEQ ID NO.13所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.30所示序列或与SEQ ID NO.30所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.35所示序列或与SEQ ID NO.35所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；或
HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.30所示序列或与SEQ ID NO.30所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID  NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.13所示序列或与SEQ ID NO.13所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；
其中HCDR和LCDR序列根据Kabat定义。
在一些实施方案中，重链可变区具有SEQ ID NO.4、18、19、20、22、24、12、25、26、28、29或31所示氨基酸序列或与SEQ ID NO.4、18、19、20、22、24、12、25、26、28、29或31所示氨基酸序列具有至少80％、至少85％、至少90％、至少95％或至少99％同一性的氨基酸序列。
在一些实施方案中，轻链可变区具有SEQ ID NO.8、33、34、16、36、37或38所示氨基酸序列或与SEQ ID NO.8、33、34、16、36、37或38所示氨基酸序列具有至少80％、至少85％、至少90％、至少95％或至少99％同一性的氨基酸序列。
在一些实施方案中，重链可变区序列为SEQ ID NO.4所示序列，轻链可变区的序列为SEQ ID NO.8所示序列；或
重链可变区序列为SEQ ID NO.18所示序列，轻链可变区的序列为SEQ ID NO.33所示序列；或
重链可变区序列为SEQ ID NO.18所示序列，轻链可变区的序列为SEQ ID NO.34所示序列；或
重链可变区序列为SEQ ID NO.19所示序列，轻链可变区的序列为SEQ ID NO.33所示序列；或
重链可变区序列为SEQ ID NO.19所示序列，轻链可变区的序列为SEQ ID NO.34所示序列；或
重链可变区序列为SEQ ID NO.20所示序列，轻链可变区的序列为SEQ ID NO.33所示序列；或
重链可变区序列为SEQ ID NO.20所示序列，轻链可变区的序列为SEQ ID NO.34所示序列；或
重链可变区序列为SEQ ID NO.22所示序列，轻链可变区的序列为SEQ ID NO.33所示序列；或
重链可变区序列为SEQ ID NO.22所示序列，轻链可变区的序列为SEQ ID NO.34所示序列；或
重链可变区序列为SEQ ID NO.24所示序列，轻链可变区的序列为SEQ ID NO.33所示序列；或
重链可变区序列为SEQ ID NO.24所示序列，轻链可变区的序列为SEQ ID NO.34所示序列；或
重链可变区序列为SEQ ID NO.12所示序列，轻链可变区的序列为SEQ ID NO.16所示序列；或
重链可变区序列为SEQ ID NO.25所示序列，轻链可变区的序列为SEQ ID NO.36所示序列；或
重链可变区序列为SEQ ID NO.25所示序列，轻链可变区的序列为SEQ ID NO.37所示序列；或
重链可变区序列为SEQ ID NO.25所示序列，轻链可变区的序列为SEQ ID NO.38所示序列；或
重链可变区序列为SEQ ID NO.26所示序列，轻链可变区的序列为SEQ ID NO.36所示序列；或
重链可变区序列为SEQ ID NO.26所示序列，轻链可变区的序列为SEQ ID NO.37所示序列；或
重链可变区序列为SEQ ID NO.26所示序列，轻链可变区的序列为SEQ ID NO.38所示序列；或
重链可变区序列为SEQ ID NO.28所示序列，轻链可变区的序列为SEQ ID NO.36所示序列；或
重链可变区序列为SEQ ID NO.28所示序列，轻链可变区的序列为SEQ ID NO.37所示序列；或
重链可变区序列为SEQ ID NO.28所示序列，轻链可变区的序列为SEQ ID NO.38所示序列；或
重链可变区序列为SEQ ID NO.29所示序列，轻链可变区的序列为SEQ ID NO.36所示序列；或
重链可变区序列为SEQ ID NO.29所示序列，轻链可变区的序列为SEQ ID NO.37所示序列；或
重链可变区序列为SEQ ID NO.29所示序列，轻链可变区的序列为SEQ ID NO.38所示序列；或
重链可变区序列为SEQ ID NO.31所示序列，轻链可变区的序列为SEQ ID NO.36所示序列；或
重链可变区序列为SEQ ID NO.31所示序列，轻链可变区的序列为SEQ ID NO.37所示序列；或
重链可变区序列为SEQ ID NO.31所示序列，轻链可变区的序列为SEQ ID NO.38所示序列。
第二方面，本申请提供了抗CD73抗体或其抗原结合部分，所述抗CD73抗体或其抗原结合部分包括重链可变区和轻链可变区，其中：
所述重链可变区具有SEQ ID NO.4、18、19、20、22、24、12、25、26、28、29或31所示氨基酸序列或与SEQ ID NO.4、18、19、20、22、24、12、25、26、28、29或31所示氨基酸序列具有至少80％、至少85％、至少90％、至少95％或至少99％同一性的氨基酸序列；和/或
所述轻链可变区具有SEQ ID NO.8、33、34、16、36、37或38所示氨基酸序列或与SEQ ID NO.8、33、34、16、36、37或38所示氨基酸序列具有至少80％、至少85％、至少90％、至少95％或至少99％同一性的氨基酸序列。
第三方面，本申请提供了抗CD73抗体或其抗原结合部分，所述抗CD73抗体或其抗原结合部分包括重链可变区，所述重链可变区包括重链CDR1(HCDR1)、重链CDR2(HCDR2)和重链CDR3(HCDR3)，其中：
(1)HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.2所示序列或与SEQ ID NO.2所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性；或
(2)HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.17所示序列或与SEQ ID NO.17所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性；或
(3)HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.21所示序列或与SEQ  ID NO.21所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性；或
(4)HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.23所示序列或与SEQ ID NO.23所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性；或
(5)HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.10所示序列或与SEQ ID NO.10所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性；或
(6)HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.27所示序列或与SEQ ID NO.27所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性；或
(7)HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.30所示序列或与SEQ ID NO.30所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性；
其中HCDR序列根据Kabat定义。
第四方面，本申请提供了抗CD73抗体或其抗原结合部分，所述抗CD73抗体或其抗原结合部分包括轻链可变区，轻链可变区包括轻链CDR1(LCDR1)、轻链CDR2(LCDR2)和轻链CDR3(LCDR3)，其中：
(1)LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.6所示序列或与SEQ ID NO.6所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
(2)LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.32所示序列或与SEQ ID NO.32所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
(3)LCDR1的序列为SEQ ID NO.13所示序列或与SEQ ID NO.13所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；或
(4)LCDR1的序列为SEQ ID NO.35所示序列或与SEQ ID NO.35所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％ 或99％同一性；
其中LCDR序列根据Kabat定义。
在第一至第四方面的一些实施方案中，抗CD73抗体或其抗原结合部分结合人CD73。在一些具体实施方案中，抗CD73抗体或其抗原结合部分与来自不同物种的CD73间(例如人和食蟹猴CD73间)保守的CD73蛋白表位结合。
在第一至第四方面的一些实施方案中，抗CD73抗体或其抗原结合部分具有ADCC活性。
在第一至第四方面的一些实施方案中，抗CD73抗体或其抗原结合部分具有CDC活性。
在第一至第四方面的一些实施方案中，抗CD73抗体为全抗体、单链抗体(scFv)、scFv-Fc、双特异性抗体或多特异性抗体。
在第一至第四方面的一些实施方案中，抗CD73抗体的抗原结合部分为Fab、Fab’、Fv或F(ab’)2。
在第一至第四方面的一些实施方案中，抗CD73抗体为鼠源抗体、嵌合抗体、人源化抗体或全人源抗体。
在第一至第四方面的一些实施方案中，抗CD73抗体为单克隆抗体。
在第一至第四方面的一些实施方案中，抗CD73抗体为IgM、IgD、IgG、IgA或IgE型抗体。在一些具体实施方案中，抗CD73抗体为IgG抗体。在一些更具体的实施方案中，抗CD73抗体为IgG1抗体。
在第一至第四方面的一些实施方案中，抗CD73抗体为IgG1、IgG2或IgG4同种型的。
在第一至第四方面的一些实施方案中，抗CD73抗体包含κ亚型或者λ亚型的轻链恒定区。
在第一至第四方面的一些实施方案中，抗CD73抗体包含人IgG1重链恒定区和人κ轻链恒定区。
在第一至第四方面的一些实施方案中，抗CD73抗体或其抗原结合部分用于医疗疗法。
在第一至第四方面的一些实施方案中，抗CD73抗体包含野生型Fc区。
在第一至第四方面的一些实施方案中，抗CD73抗体或其抗原结合部分经过工程化改造具有增强的ADCC或CDC活性。在一些具体实施方案中，抗CD73抗体包含经过工程改造的Fc区，所述经过工程改造的Fc区使得所述抗体具有增强的ADCC和/或CDC效应。
在第一至第四方面的一些实施方案中，抗CD73抗体或其抗原结合部分经过工程化改造具有减弱的ADCC或CDC活性。在一些具体实施方案中，抗CD73抗体包含经过工程改造的Fc区，所述经过工程改造的Fc区使得所述抗体具有减弱的ADCC和/或CDC效应。针对本申请针对的靶点CD73以及抗体的体内应用，具有减弱的、甚至消除的ADCC或CDC活性的抗体可能是有利的，因为过强的Fc效应可能会影响甚至消弱抗体的疗效。减弱或者消除Fc效应的方式是本领域中已知的，例如，1)氨基酸残基突变改造(主要减弱对相关受体的结合)，2)糖基化改造，3)IgG4型抗体改造等。作为非限制性实例，抗体具有人IgG1重链恒定区，并且含有L234、L235、P329、P331中一个或多个位点的突变，其中残基编号按照EU编号***，例如，所述突变为L234A、L235A、P329A、P331S中的一种或多种突变。
在第一至第四方面的一些实施方案中，所述抗体具有SEQ ID NO.20所示的重链可变区，SEQ ID NO.33所示的轻链可变区，SEQ ID NO.39所示的重链恒定区和SEQ ID NO.40所示的轻链恒定区。
在第一至第四方面的一些实施方案中，所述抗体具有SEQ ID NO.31所示的重链可变区，SEQ ID NO.38所示的轻链可变区，SEQ ID NO.39所示的重链恒定区和SEQ ID NO.40 所示的轻链恒定区。
第五方面，本申请提供了分离的核酸分子，其编码第一方面至第四方面所述的抗CD73抗体或其抗原结合部分。在一些实施方案中，本申请提供了分离的多核苷酸的组合，所述组合包括编码本申请抗体或其抗原结合部分的轻链的多核苷酸和编码本申请抗体或其抗原结合部分的重链的多核苷酸。在一些实施方案中，所述多核苷酸可操作地连接到调控序列，调控序列可以被用所述载体转化过的宿主细胞识别。
第六方面，本申请提供了载体(例如表达载体)，其含有第五方面所述的核酸分子或多核苷酸的组合。
在一些实施方案中，本申请的表达载体包含本申请所述的核酸分子或者本申请所述的多核苷酸的组合，所述多核苷酸与允许其所编码的多肽在宿主细胞或无细胞表达***中表达的调节序列有效连接。表达载体的选择取决于宿主细胞的选择，并且可以选择以便在所选宿主细胞中具有所需的表达和调控特征。
“表达载体”是包括一个或多个表达控制序列的载体，“表达控制序列”是控制和调节另一DNA序列的转录和/或翻译的DNA序列。
载体中的核酸可以与一个或多个表达控制序列可操作地连接。如本文所用，“可操作地连接”意指并入遗传构建体中，使得表达控制序列有效地控制目标编码序列的表达。表达控制序列的示例包括启动子、增强子和转录终止区。启动子是由DNA分子的通常在转录起始点上游100个核苷酸内(通常在RNA聚合酶II的起始位点附近)的区域组成的表达控制序列。为了使编码序列处于启动子的控制下，必须将多肽翻译阅读框的翻译起始位点定位在启动子下游的1至约50个核苷酸之间。增强子在时间、位置和水平方面提供表达特异性。与启动子不同，增强子可以在位于距转录位点的不同距离时起作用。增强子也可位于转录起始位点的下游。当RNA聚合酶能够将编码序列转录成mRNA，然后mRNA可以翻译成由编码序列所编码的蛋白质时，编码序列与细胞中的表达控制序列“可操作地连接”且处于表达控制序列的“控制下”。
合适的表达载体包括但不限于源自例如噬菌体、杆状病毒、烟草花叶病毒、疱疹病毒、巨细胞病毒、逆转录病毒、痘苗病毒、腺病毒和腺相关病毒的质粒和病毒载体。许多载体和表达***可从诸如Novagen(Madison,WI)、Clontech(Palo Alto,CA)、Stratagene(LaJolla,CA)和Invitrogen Life Technologies(Carlsbad,CA)等公司商购获得。
表达载体可包括标签序列。标签序列通常表达为与编码的多肽的融合体。此类标签可以***多肽内的任何位置，包括羧基或氨基末端。有用标签的示例包括但不限于Fc片段、聚组氨酸、绿色荧光蛋白(GFP)、谷胱甘肽S-转移酶(GST)、c-myc、血凝素、FlagTM标签(Kodak,NewHaven,CT)、麦芽糖E结合蛋白和蛋白A。在一些实施方案中，编码CD73融合多肽的核酸分子存在于含有编码Ig重链恒定区的一个或多个结构域的核酸的载体中，所述结构域例如，对应与人免疫球蛋白Cγ1链的铰链区、CH 2区和CH3区的氨基酸序列(Fc片段)。
第七方面，本申请提供了宿主细胞，其含有第五方面所述的核酸分子或第六方面所述的载体。在一些实施方案中，宿主细胞可以为原核宿主细胞、真核宿主细胞或噬菌体。原核宿主细胞可以为大肠杆菌、枯草杆菌、链霉菌或奇异变形菌等。真核宿主细胞可以为如巴斯德毕赤酵母、酿酒酵母、裂殖酵母、木霉等真菌，如草地粘虫等昆虫细胞，如烟草等植物细胞，如BHK细胞、CHO细胞、COS细胞、骨髓瘤细胞等哺乳动物细胞。在一些实施方案中，宿主细胞优选为哺乳动物细胞，更优选为BHK细胞、CHO细胞、NSO细胞或COS细胞。
第八方面，本申请提供了抗体药物偶联物，也称为抗体-药物缀合物，其包含与治疗剂偶联的第一方面至第四方面所述的抗CD73抗体或其抗原结合部分。
在第八方面的一些实施方案中，治疗剂为抗肿瘤药物，例如，细胞毒性药物、免疫增强剂或放射性同位素。
在一些实施方案中，细胞毒性药物的类型包括微管蛋白抑制剂(例如生物碱)、DNA拓扑异构酶抑制剂、DNA损伤剂、抗代谢药或抗肿瘤抗生素。
在一些实施方案中，微管蛋白抑制剂包括但不限于，奥瑞斯他汀衍生物(例如MMAE(Monomethyl auristatin E)、MMAF(Monomethyl auristatin F))或美登素生物碱衍生物(例如DM1、DM4、安丝菌素(Ansamitocin)、美登素(Mertansine)或海兔毒素肽(dolastatin)及其衍生物)。
在一些实施方案中，DNA拓扑异构酶抑制剂为喜树碱类似物或DNA拓扑异构酶I抑制剂及其衍生物，例如，DXD、SN38、伊立替康、伊立替康盐酸盐、喜树碱、9-氨基喜树碱、9-硝基喜树碱、10-羟基喜树碱、9-氯-10-羟基喜树碱、22-羟基旱莲木碱、拓扑替康、勒托替康、贝洛替康、依喜替康、硅基高喜树碱(homosilatecan)、6,8-二溴-2-甲基-3-[2-(D-吡喃木糖基氨基)苯基]-4(3H)-喹唑啉酮、2-氰基-3-(3,4-二羟基苯基)-N-(苯基甲基)-(2E)-2-丙烯酰胺、2-氰基-3-(3,4-二羟基苯基)-N-(3-羟基苯基丙基)-(E)-2-丙烯酰胺、12-β-D-吡喃萄萄糖基-12,13-二氢-2,10-二羟基-6-[[2-羟基-1-(羟基甲基)乙基]氨基]-5H-吲哚并[2,3-a]吡咯并[3,4-c]咔唑-5,7(6H)-二酮、N-[2-(二甲基氨基)乙基]-4-吖啶甲酰胺二盐酸盐、N-[2-(二甲基氨基)乙基]-4-吖啶甲酰胺。
在一些实施方案中，DNA损伤剂包括但不限于，卡奇霉素(calicheamicin)类、倍癌霉素(duocarmycin)类、安曲霉素类衍生物PBD(pyrrolobenzodiazepine，吡咯并苯并二氮杂)。
在一些实施方案中，抗代谢药包括但不限于，甲氨蝶呤、6-巯嘌呤或5-氟尿嘧啶。
在一些实施方案中，抗肿瘤抗生素包括但不限于，多肽类抗生素(例如放线菌素D或博来霉素)或蒽醌类药物(例如阿霉素或盐酸米托蒽醌)。在一些实施方案中，放射性同位素包括但不限于，211At、131I、125I、90Y、186Re、188Re、153Sm、212Bi、32P、60Co或177Lu。
在一些实施方案中，免疫增强剂包括但不限于，左旋咪唑(levamisole)、匹多莫德(pidotimod)、咪喹莫特(imiquimod)、异丙肌苷、聚肌胞苷酸或聚肌尿苷酸。
在一些实施方案中，本申请的抗CD73抗体通过接头与药物部分共价连接。在一些实施方案中，接头是可切割接头。在一些实施方案中，接头在细胞内条件下可被切割。在一个实施方案中，接头在小于5.5的pH下可水解。在一些实施方案中，接头可被细胞内蛋白酶切割。在一些实施方案中，接头是组织蛋白酶可切割接头。在一些实施方案中，接头包含二肽。在一些实施方案中，二肽是缬氨酸(Val)-瓜氨酸(Cit)。在一些实施方案中，抗体通过抗体的半胱氨酸巯基与接头连接。在一个实施方案中，抗体通过抗体的氨基(特别是谷氨酰胺残基的氨基)与接头连接。接头的非限制性实例包括mc-Val-Cit-pAB、mc-Val-Cit-pABC、mc-Val-Cit、NH2-(PEG)m-Val-Cit、NH2-(PEG)m-Val-Cit-pAB，其中m为1至8的整数。在一些实施方案中，接头是非可切割接头，包括而不限于，酸不稳定接头(例如腙接头)、含二硫键的接头、肽酶敏感的接头(例如包含氨基酸的肽接头，例如缬氨酸和/或瓜氨酸例如瓜氨酸-缬氨酸或苯丙氨酸-赖氨酸)、光不稳定接头、二甲基接头、硫醚接头或经设计避免多药转运蛋白介导的抗性的亲水接头等。
第九方面，本申请提供了药物组合物，其包含第一方面至第四方面所述的抗CD73抗体或其抗原结合部分或第八方面所述的抗体药物偶联物，以及药学可接受的载体。
在一些实施方案中，药物组合物用于治疗个体中的肿瘤。
在一些实施方案中，肿瘤为肿瘤细胞表面表达CD73(CD73+)的肿瘤。
在一些实施方案中，肿瘤为肿瘤细胞表面高表达CD73(CD73+)的肿瘤。在一些实施方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少60％的肿瘤细胞表达CD73。在一些实施方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少70％的肿瘤细胞表达CD73。在一些实施方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少80％的肿瘤细胞表达CD73。在一些实施方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少90％的肿瘤细胞表达CD73。在一些实施方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少95％的肿瘤细胞表达CD73。在一些实施 方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少98％的肿瘤细胞表达CD73。在一些实施方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少99％的肿瘤细胞表达CD73。
在一些实施方案中，肿瘤为实体瘤。
在一些实施方案中，肿瘤为血液肿瘤。
在一些实施方案中，肿瘤为恶性肿瘤。
在一些实施方案中，肿瘤为癌症。
在一些实施方案中，肿瘤选自：卵巢癌、乳腺癌、黑色素瘤、结直肠癌、胃癌、肾癌、***癌、胰腺癌、食管癌、头颈癌、甲状腺癌、肺癌、膀胱癌、***、结肠癌、肝癌、腹膜癌、肝细胞癌、成胶质细胞瘤、尿路癌、直肠癌、子宫内膜癌、子宫癌、唾液腺癌、鳞状细胞癌、外阴癌、***癌、***癌、脑癌、淋巴瘤(例如B细胞淋巴瘤)、白血病，以及上述肿瘤的转移灶。
在一些实施方案中，药物组合物还可包含下述中的一种或多种：润滑剂，如滑石粉、硬脂酸镁和矿物油；润湿剂；乳化剂；悬浮剂；防腐剂，如苯甲酸、山梨酸和丙酸钙；增甜剂和/或调味剂等。在一些实施方案中，可将本申请中的药物组合物配制为片剂、丸剂、粉剂、锭剂、酏剂、悬液、乳剂、溶液、糖浆、栓剂或胶囊等形式。
在一些实施方案中，可以利用任何生理上可接受的给药方式递送本申请的药物组合物，这些给药方式包括但不限于：口服给药、肠胃外给药、经鼻给药、直肠给药、腹膜内给药、血管内注射、皮下给药、经皮给药、吸入给药等。
在一些实施方案中，可以通过混合具有所需纯度的试剂与视情况的药学上可接受的载体、赋形剂等，以冻干制剂或水溶液的形式配制用于治疗用途的药物组合物用于存储。
第十方面，本申请提供了第一方面至第四方面所述的抗CD73抗体或其抗原结合部分、第五方面所述的核酸分子、第六方面所述的载体、第七方面所述的宿主细胞或第八方面所述的抗体药物偶联物在制备用于治疗个体中的肿瘤，特别是肿瘤细胞表面表达CD73(CD73+)的肿瘤的药物中的用途。
在一些实施方案中，肿瘤为肿瘤细胞表面表达CD73(CD73+)的肿瘤。
在一些实施方案中，肿瘤为肿瘤细胞表面高表达CD73(CD73+)的肿瘤。在一些实施方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少60％的肿瘤细胞表达CD73。在一些实施方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少70％的肿瘤细胞表达CD73。在一些实施方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少80％的肿瘤细胞表达CD73。在一些实施方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少90％的肿瘤细胞表达CD73。在一些实施方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少95％的肿瘤细胞表达CD73。在一些实施方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少98％的肿瘤细胞表达CD73。在一些实施方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少99％的肿瘤细胞表达CD73。
在一些实施方案中，肿瘤为实体瘤。
在一些实施方案中，肿瘤为血液肿瘤。
在一些实施方案中，肿瘤为恶性肿瘤。
在一些实施方案中，肿瘤为癌症。
在一些实施方案中，肿瘤选自：卵巢癌、乳腺癌、黑色素瘤、结直肠癌、胃癌、肾癌、***癌、胰腺癌、食管癌、头颈癌、甲状腺癌、肺癌、膀胱癌、***、结肠癌、肝癌、腹膜癌、肝细胞癌、成胶质细胞瘤、尿路癌、直肠癌、子宫内膜癌、子宫癌、唾液腺癌、鳞状细胞癌、外阴癌、***癌、***癌、脑癌、淋巴瘤(例如B细胞淋巴瘤)、白血病，以及上述肿瘤的转移灶。
第十一方面，本申请提供了治疗个体中的肿瘤，特别是肿瘤细胞表面表达CD73 (CD73+)的肿瘤的方法，所述方法包括给予所述个体治疗有效量的第一方面至第四方面所述的抗CD73抗体或其抗原结合部分、第八方面所述的抗体药物偶联物或第九方面所述的药物组合物。在治疗或预防疾病、病症或病况的方法的一些实施方案中，方法包括将细胞体外暴露于抗CD73抗体中。
在一些实施方案中，肿瘤为肿瘤细胞表面表达CD73(CD73+)的肿瘤。
在一些实施方案中，肿瘤为肿瘤细胞表面高表达CD73(CD73+)的肿瘤。在一些实施方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少60％的肿瘤细胞表达CD73。在一些实施方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少70％的肿瘤细胞表达CD73。在一些实施方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少80％的肿瘤细胞表达CD73。在一些实施方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少90％的肿瘤细胞表达CD73。在一些实施方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少95％的肿瘤细胞表达CD73。在一些实施方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少98％的肿瘤细胞表达CD73。在一些实施方案中，高表达CD73(CD73+)的肿瘤是指肿瘤细胞群体中至少99％的肿瘤细胞表达CD73。
在一些实施方案中，肿瘤为实体瘤。
在一些实施方案中，肿瘤为血液肿瘤。
在一些实施方案中，肿瘤为恶性肿瘤。
在一些实施方案中，肿瘤为癌症。
在一些实施方案中，肿瘤选自：卵巢癌、乳腺癌、黑色素瘤、结直肠癌、胃癌、肾癌、***癌、胰腺癌、食管癌、头颈癌、甲状腺癌、肺癌、膀胱癌、***、结肠癌、肝癌、腹膜癌、肝细胞癌、成胶质细胞瘤、尿路癌、直肠癌、子宫内膜癌、子宫癌、唾液腺癌、鳞状细胞癌、外阴癌、***癌、***癌、脑癌、淋巴瘤(例如B细胞淋巴瘤)、白血病，以及上述肿瘤的转移灶。
在第九至十一方面的一些实施方案中，本申请的抗CD73抗体或其抗原结合部分可单独使用或与抗肿瘤疗法中的其它组合物联用。例如，抗CD73抗体可与至少一种另外的治疗剂和/或佐剂共同给予。在一些实施方案中，另外的化合物是抗CD73抗体以外的治疗性抗体。在一些实施方案中，其它治疗剂是靶向癌症免疫周期的治疗剂(例如：免疫检查点抑制剂(例如，抗PD-1抗体、抗PD-L1抗体或抗CTLA4抗体)，A2AR拮抗剂，STAT-3抑制剂)，使用这样的治疗剂联合疗法可用于降低肿瘤介导的免疫抑制。
第十二方面，本申请提供了缀合物，其包含第一方面至第四方面所述的抗CD73抗体或其抗原结合部分以及可检测标记物。可检测标记物例如可检测的荧光标记物、化学发光标记物、同位素标记物等。
第十三方面，本申请提供了包含第一方面至第四方面所述的抗CD73抗体或其抗原结合部分的融合蛋白。抗体融合蛋白的实例包括Fab融合蛋白、Fc融合蛋白与单链抗体(scFv)融合蛋白等，根据效应蛋白(例如细胞因子)所融合的部位不同而命名。
在十二或十三方面的一些实施方案中，可以通过合成接头与一种或多种非抗体作用剂共价结合的本申请的抗CD73抗体而形成缀合物。在一些实施方案中，使本申请的抗体与诊断剂、检测剂或治疗剂或任何其它分子缀合或重组融合。缀合或重组融合的抗体可用于例如监测或预测CD73介导的疾病的发作、发生、进展和/或严重程度作为临床试验法的一部分，例如测定特定疗法的功效。这类诊断和检测可通过例如使抗体与可检测标记物偶联来实现。可检测标记物包括但不限于酶、辅基、荧光标记物、化学发光标记物、同位素标记物等。在一些实施方案中，本申请的抗体可与修饰指定生物应答的治疗部分或药物部分缀合或重组融合(此处也可能是第八方面所述的抗体药物偶联物)。治疗部分或药物部分不限于典型的化学治疗剂。例如，药物部分可以是具有所需生物活性的蛋白质、肽、多肽、小分子毒素等。
应当理解，以上详细描述仅为了使本领域技术人员更清楚地了解本申请的内容，而并非意图在任何方面加以限制。本领域技术人员能够对所述实施方案进行各种改动和变化。
实施例
下文将结合实施例对本申请的具体实施方案进行详细描述，但是本领域技术人员将会理解，以下实施例仅用于说明本申请的发明，而不应视为限制本申请的范围。
实施例1：抗人CD73单克隆抗体的制备
使用重组人CD73蛋白(Arco，货号CD3-H52H7)免疫健康的BALB/c小鼠，初次免疫后，每隔14天进行加强免疫3次。取小鼠外周血清，通过酶联免疫法(ELISA)检测小鼠血清效价，选择血浆抗体滴度高的小鼠进行细胞融合。融合前3天通过尾静脉或腹腔进行冲击免疫。融合当天取小鼠脾脏，制备成单细胞悬液，将SP2/0细胞与脾细胞按1:2的比例混合，通过电融合的方法进行融合。置于37℃CO2培养箱中培养，当杂交瘤细胞生长至一定量时，通过ELISA方法对杂交瘤上清进行筛选。挑选出阳性克隆，通过有限稀释法进行亚克隆筛选。最终将获得的单克隆阳性细胞通过提取RNA、反转录及PCR扩增，获得鼠免疫球蛋白重链和轻链可变区片段，进行测序。最终获得了7C3C2、1F2F9克隆的重链和轻链CDR区序列和可变区的氨基酸序列如下：
7C3C2
重链CDR1：SYDIN(SEQ ID NO.1)
重链CDR2:WIYPRDGFTKYNEKFKG(SEQ ID NO.2)
重链CDR3:KEGYRGDWYFDV(SEQ ID NO.3)
重链可变区序列
轻链CDR1：RASGNVHNYLA(SEQ ID NO.5)
轻链CDR2：NAKTLAD(SEQ ID NO.6)
轻链CDR3：QHFWSTPWT(SEQ ID NO.7)
轻链可变区序列
1F2F9
重链CDR1：SYWMH(SEQ ID NO.9)
重链CDR2:NINPSNGGTKYNEKFKS(SEQ ID NO.10)
重链CDR3:RDYGSPSY(SEQ ID NO.11)
重链可变区序列
轻链CDR1：KASQDINSYLT(SEQ ID NO.13)
轻链CDR2：RTNILLD(SEQ ID NO.14)
轻链CDR3：LQYHEFPVT(SEQ ID NO.15)
轻链可变区序列

注：上述抗体可变区序列中，下划线为CDR序列(根据Kabat编号***确定并注释)。
实施例2：ELISA法检测嵌合抗体的亲和力
根据上述单克隆抗体编码基因的可变区序列信息，构建人鼠嵌合抗体表达载体。将VH和VL分别构建到含有hIgG1-kappa恒定区的真核表达载体上，其中含有L234A、L235A、P329A、P331S突变(本申请中简称为hIgG1，重链恒定区序列如SEQ ID NO.39所示，轻链恒定区序列如SEQ ID NO.40所示，下文实施例与此相同)。
将获得的真核表达载体，瞬转入HEK293细胞中，培养6天。收获培养上清通过ProteinA柱进行纯化，获得目标抗体，分别命名为7C3C2-hIgG1、1F2F9-hIgG1，采用ELISA法测定抗体与重组人CD73蛋白(同实施例1)的亲和力。将重组人CD73蛋白按1μg/ml的浓度包被酶标板，PBST缓冲液洗板3次；然后用含2％BSA的PBS缓冲液，37℃封闭2h，PBST缓冲液洗板3次；加入5倍梯度稀释的待测抗体(0-133.3nM)，37℃孵育2h，PBST缓冲液洗板3次。加入辣根过氧化物酶标记的二抗，37℃孵育1h，PBST缓冲液洗板3次。显色，终止后读取吸光值。所得亲和力如图1及表1所示。
表1.嵌合抗体亲和力的EC50值
抗体名称 EC50值(nM)
7C3C2-hIgG1 0.017
1F2F9-hIgG1 0.028
7C3C2-hIgG1和1F2F9-hIgG1与CD73蛋白结合的EC50分别为0.017nM及0.028nM。由此可见，这两株抗体的EC50值满足进一步实验的要求。
实施例3：抗体人源化
使用Discovery Studio和Antibody Modeling进行同源建模。通过结构模拟与理性设计，分析并获得与鼠源抗体框架区最接近的人源框架区。对于鼠源7C3C2的人源化轻链模板为IGKV1-NL1；对于重链来说，最接近的人类匹配序列是IGHV1-8基因；鼠源1F2F9的人源化轻链模板为IGKV1-16基因，最接近的人类重链匹配序列是IGHV1-3基因。将轻重链的CDR分别移植到匹配的轻链和重链基因的框架序列上，即获得人源化抗体Hab7C3C2及Hab1F2F9。然后通过Discovery Studio和Antibody Modeling构建3D模型，分析是否存在任何将小鼠氨基酸取代为人氨基酸的框架位置影响结合和/或CDR构象的位点，进行回复突变序列见表2及表3。相关人源化抗体见表4及表5。


表4：7C3C2人源化抗体组合示意图
  Hab7C3C2-VL.1 Hab7C3C2-VL.1a
Hab7C3C2-VH.1 Hab7C3C2.1 Hab7C3C2.2
Hab7C3C2-VH.1a Hab7C3C2.3 Hab7C3C2.4
Hab7C3C2-VH.1b Hab7C3C2.5 Hab7C3C2.6
Hab7C3C2-VH.1c Hab7C3C2.7 Hab7C3C2.8
Hab7C3C2-VH.1d Hab7C3C2.9 Hab7C3C2.10
表5：1F2F9人源化抗体组合示意图
  Hab1F2F9-VL.1 Hab1F2F9-VL.1a Hab1F2F9-VL.1b
Hab1F2F9-VH.1a Hab1F2F9.1 Hab1F2F9.2 Hab1F2F9.3
Hab1F2F9-VH.1b Hab1F2F9.4 Hab1F2F9.5 Hab1F2F9.6
Hab1F2F9-VH.1c Hab1F2F9.7 Hab1F2F9.8 Hab1F2F9.9
Hab1F2F9-VH.1d Hab1F2F9.10 Hab1F2F9.11 Hab1F2F9.12
Hab1F2F9-VH.1e Hab1F2F9.13 Hab1F2F9.14 Hab1F2F9.15
实施例4：人源化抗CD73抗体亲和力检测
4.1 Fortebio检测人源化抗体与人CD73蛋白的亲和力
将人源化抗体VH和VL分别构建到含有hIgG1-kappa恒定区的真核表达载体上，其中含有L234A、L235A、P329A、P331S突变(本申请中简称为hIgG1)，本实施例检测了所有人源化抗体的亲和力。使用HIS1K生物传感器，将人的CD73蛋白固定在生物传感器上，至0.3nm的结合响应阈值。在120s基线步骤后，将传感器分别浸入用0.02％PBST缓冲液梯度稀释的抗CD73抗体分析物中，抗体起始浓度为25nM，2倍梯度稀释，设置7个浓度梯度。配体与分析物结合和解离时间分别为120s和300s。根据结合曲线由分析软件，回归计算出抗原抗体的亲和力数据。在所有检测抗体中Hab7C3C2.5、Huab7C3C.9、Huab1F2F9.12及Huab1F2F9.15表现出最高的亲和力和较少的回复突变，结果如表6所示。
表6：Fortebio法检测人源化抗体与人CD73蛋白的亲和力
测试物 Ka(1/Ms) Kd(1/s) KD(M)
Hab7C3C2.5 3.56E06 2.72E-03 7.653E-10
Hab7C3C2.9 3.7E06 3.81E-03 1.03E-09
Hab1F2F9.12 1.85E06 1.33E-03 7.195E-10
Hab1F2F9.15 2.06E06 1.6E-03 7.787E-10
4.2 ELISA检测人源化抗体与人CD73蛋白的亲和力
采用ELISA法测定人源化抗体与重组CD73蛋白的亲和力，将CD73蛋白按1μg/ml的浓度包被酶标板；加入5倍梯度稀释的抗体(0-66.7nM)，抗体所得亲和力如图2及表7所示。
表7.ELISA法检测人源化抗体亲和力的EC50值
测试物 EC50值(nM)
Hab7C3C2.5 0.013
Hab1F2F9.15 0.011
Hab7C3C2.5及Huab1F2F9.15与CD73蛋白结合的EC50分别为0.013nM及0.011nM。 由此可见，人源化后的抗体与鼠源抗体相比，亲和力未受到影响。
实施例5：人源化抗体与人源肿瘤细胞的结合
本实施例将本申请的示例性抗体与其它专利申请中描述的抗CD73抗体进行比较。将MEDI9447及Hu101-28(CDR区序列分别来自PCT专利申请WO 2016/075099及WO2018/137598)抗体克隆至含有hIgG1-kappa恒定区的真核表达载体上。使用细胞表面内源表达人CD73的肿瘤细胞系MDA-MB-231及SKOV3，评估人源化抗体Hab7C3C2.5及Hab1F2F9.15与细胞表面CD73的结合能力。将细胞按照2×106/ml的密度，100μl/孔接种于96孔板。加入4倍梯度稀释的抗体(0-100nM)，4℃孵育1h。加入山羊抗人二抗，4℃避光标记50min，通过流式细胞仪读取细胞的荧光强度。实验结果如图3、4及表8所示，结果表明：本申请的示例性人源化抗体可以剂量依赖性结合细胞系上表达的CD73蛋白。同时，Hab7C3C2.5和Hab1F2F9.15在识别细胞表面CD73的能力上优于对照分子Hu101-28，与MEDI9447相当。
表8本申请的示例性人源化抗体与人肿瘤细胞表面上CD73结合实验
实施例6：人源化抗CD73抗体与食蟹猴CD73的交叉反应
6.1 Fortebio法检测人源化抗体与猴CD73蛋白交叉识别
本实施例将本申请的示例性人源化抗CD73抗体与食蟹猴CD73的交叉结合力通过Fortebio实验进行检测。具体实施步骤包括将食蟹猴的CD73蛋白固定在HIS1K生物传感器上。在120s基线步骤后，将传感器分别浸入用0.02％PBST缓冲液梯度稀释的抗CD73抗体分析物中，抗体起始浓度为25nM，分别设置7个浓度梯度，进行亲和力分析实验。配体与分析物结合和解离时间分别为120s和300s。根据结合曲线由分析软件回归计算出抗原抗体的亲和力数据。结果如表9所示，本申请的示例性抗体可交叉识别食蟹猴的CD73蛋白。
表9：Fortebio法检测人源化抗体与食蟹猴CD73蛋白的亲和力
测试物 Ka(1/Ms) Kd(1/s) KD(M)
Hab7C3C2.5 4.08E06 2.99E-03 7.32E-10
Hab7C3C2.9 4.56E06 3.88E-03 8.52E-10
Hab1F2F9.12 2.22E06 1.52E-03 6.81E-10
Hab1F2F9.15 2.09E06 1.7E-03 8.14E-10
6.2流式细胞术检测人源化抗体与猴CD73蛋白的交叉识别
为了表征本申请制备的人源化抗体对细胞膜上食蟹猴CD73分子的识别能力，构建了可以稳定表达猴源全长CD73的CHO-K1细胞。将细胞以2×106/ml的密度，100μl/孔接种于96孔板。用PBS缓冲液洗三次，弃上清。加入PBS缓冲液梯度稀释的抗体，4℃孵育1h。用PBS缓冲液洗涤细胞3次，随后用Alexa Fluor 488标记的山羊抗人IgG抗体在4℃避光标记50min。用PBS缓冲液洗涤细胞3次，通过流式细胞仪读取细胞的荧光强度。
如图5及表10结果所示，制备的人源化抗体可以剂量依赖性结合细胞上表达的猴 CD73蛋白。
表10本申请的示例性人源化抗体与细胞表面上食蟹猴CD73结合实验
测试物 细胞水平亲和力EC50(nM)
Hab7C3C2.5 11.34
Hab1F2F9.15 1.51
实施例7：抗CD73抗体介导的CD73酶活抑制能力
7.1 CD73蛋白酶活抑制
将重组人CD73蛋白稀释至终浓度0.5μg/ml，加入4倍梯度稀释(0-5nM)的抗CD73抗体，以人IgG1为阴性对照，37℃孵育1h。加入ATP(75μM)和AMP(94μM)至每孔中，37℃孵育1h。将等体积的含有荧光素酶的CellTiter-Glo加入每孔中，通过酶标仪读取自发光数值。结果如图6及表11所示：Hab7C3C.5及Hab1F2F9.15可抑制游离CD73蛋白的酶活性。
表11人源化抗体对CD73蛋白酶活抑制活性数据
测试物 蛋白酶活抑制IC50(nM)
Hab7C3C2.5 0.018
Hab1F2F9.15 0.011
7.2人MDA-MB-231细胞上的CD73酶活抑制活性
在50μl/孔不同剂量的3倍梯度稀释(0-10nM)的抗CD73抗体或同种型对照抗体(人IgG1)存在的情况下，将表达CD73的MDA-MB-231细胞以2×105/ml，100μl/孔接种到96孔板中。将板在37℃孵育1h，然后每孔加入AMP(1800μM)，50μl/孔，于37℃孵育2小时。以1000rpm 5min离心细胞，并将50μl上清液转移至新的96孔板，加入50μl ATP使其终浓度为75μM。以1：1比例加入CellTiter-Glo试剂(Promega)，测定细胞性的CD73酶活性。结果如图7及表12所示：Hab7C3C.5及Hab1F2F9.15可抑制MDA-MB-231细胞上的CD73酶活性。
表12人源化抗体对MDA-MB-231细胞上的CD73酶活抑制活性
测试物 细胞酶活抑制IC50(nM)
Hab7C3C2.5 0.027
Hab1F2F9.15 0.069
实施例8：抗体诱导细胞表面CD73的内化
在本实施例中，通过流式细胞术，评估抗体介导的CD73的内化。将细胞悬浮于无血清DMEM培养基中，在冰上分别与所检验抗体一起孵育1h，抗体浓度为10μg/ml。用无血清DMEM培养基洗涤细胞三次；将孵育不同抗体的细胞分别悬浮于无血清DMEM培养基中，各分为两份分别铺于两个24孔板中。一块培养板置37℃培养箱中温育；另一培养板置4℃冰箱，作为对照组；培养24h。收集并洗涤细胞，添加Alexa Fluor 488标记的山羊抗人二抗，在4℃避光染色1h。在PBS缓冲液中洗涤3次，通过流式细胞仪进行检测。
结果图8所示，本申请的示例性抗体可诱导约60％的CD73受体内化，保持40％左右的初始表面染色。所有计算值为与该对照(设定为100％)相比的内化百分比。
实施例9：抗体的识别表位鉴定
在本实施例中，分别构建了CD73蛋白的C端部分(SEQ ID NO.41)以及N端部分(SEQ ID NO.42)，通过ELISA法检测抗体识别表位。将CD73N端及C端的蛋白以1μg/ml的浓度分别包被酶标板，梯度稀释待测抗体。所得亲和力见图9A(N端)、9B(C端)。实验结果表明：与对照抗体(MEDI9447和Hu101-28)相比，Hab7C3C.5及Hab1F2F9.15的结合表位存在差别；Hab7C3C.5及Hab1F2F9.15不结合CD73蛋白的N端和C端，MEDI9447结合CD73蛋白N端，Hu101-28结合CD73蛋白C端。本申请的Hab7C3C.5及Hab1F2F9.15的结合表位与MEDI9447和Hu101-28明显不同。
实施例10：抗CD73抗体的动物体内生物学活性评价
PBMC免疫重建鼠中评价嵌合抗体单药的药效
在肿瘤异种移植模型中评估了1F2F9嵌合抗体的单药治疗功效。将4×106个A375黑素瘤细胞和4×105个人PBMC与基质胶1：1混合后接种到NCG免疫缺陷小鼠中。在移植当天，通过腹膜内注射30mg/kg的1F2F9嵌合抗体或PBS对照，并每隔一天注射一次。治疗后24天，多只小鼠肿瘤大小达到上限(2000mm3)。因此治疗后24天成为分析的数据终点。在对照组中，平均肿瘤体积在治疗24天为1448±176.76mm3(平均值±标准误差)。与对照组相比，1F2F9-hIgG1及MEDI9447的给药减小了肿瘤生长，分别造成31.14％和16.88％的肿瘤抑制，肿瘤体积分别为997.37±131.39mm3和1203.95±158.01mm3(图10A)。单个肿瘤体积的结果显示在图10B(多条线分别代表多只小鼠)中。各实验组的体重变化并不显著(图10C)，表明治疗得到了良好的耐受。
实施例11：基因鼠中评价CD73人源化抗体的药效
在本实施例中，利用人CD73/PD1/PDL1敲入的BALB/c小鼠和过表达人CD73/PDL1的CT26细胞(结肠癌细胞系)评价Huab1F2F9.15、抗PD-1抗体以及它们组合的药效。将1×106个CT26-hCD73/hPDL1细胞皮下接种至基因鼠中。当肿瘤平均体积在60-80mm3左右，根据肿瘤体积进行分组。分组当天定义为D0天，并于分组当天开始给药。腹腔每周两次注射指定剂量的抗体或PBS对照。采用单独的抗PD-1抗体(1mg/kg)、单独的抗CD73抗体(Huab1F2F9.15及MEDI9447；15mg/kg)以及抗PD-1抗体加上抗CD73抗体(1mg/kg+15mg/kg)处理小鼠。结果表明：Huab1F2F9.15与抗PD-1抗体及MEDI9447与抗PD1抗体联合给药减小肿瘤生长，分别产生53.74％和37.67％的肿瘤抑制，Huab1F2F9.15与抗PD-1抗体联合效果显著优于MEDI9447与抗PD1抗体的组合(图11A)。单个肿瘤体积的结果显示在图11B(多条线分别代表不同小鼠)中。各实验组的体重变化并不显著(图11C)，表明治疗得到了良好的耐受性。
综合以上实验结果，本申请获得了具有高亲和力的识别CD73蛋白及细胞水平上的CD73的抗体。所测试的代表性抗体可强效的抑制CD73的酶活，同时可诱导肿瘤细胞上的CD73蛋白内化，通过降低细胞表面的CD73分子进一步降低CD73的酶活，由此实现通过多种途径抑制CD73酶活性的效果。通过表位分析显示本申请的示例性抗体对CD73蛋白单独的N端及C端蛋白都不结合，与诸如MEDI9447和Hu101-28的现有技术抗体具有差异化的表位结合特征。动物体内实验证实1F2F9嵌合抗体及人源化Huab1F2F9.15抗体可有效的抑制动物体内肿瘤细胞的增殖。特别是人源化Huab1F2F9.15抗体与抗PD-1抗体联合使用实现了肿瘤抑制的优异效果，并显示良好的耐受性。
本文提供的任何和所有实施例或示例性语言(例如，“诸如”)的使用仅旨在更好地说明本申请，而不对本申请的范围构成限制，除非另有要求。说明书中的语言不应被解释为指示任何未要求保护的元件对于实施本申请是必要的。
本说明书中引用的所有出版物和专利申请通过引用并入本文，如同每个单独的出版物或专利申请被具体地和单独地指明通过引用并入。此外，本文所述的任何理论、机制、证明或发现旨在进一步增强对本申请的理解，并且不意图以任何方式将本申请限制到这样的理论、机制、证明或发现。尽管已经在附图和前面的描述中详细地示出和描述了本申请，但是本申请应当被认为是说明性的而不是限制性的。

Claims (43)

  1. 抗CD73抗体或其抗原结合部分，所述抗CD73抗体或其抗原结合部分包括重链可变区和轻链可变区，所述重链可变区包括重链CDR1(HCDR1)、重链CDR2(HCDR2)和重链CDR3(HCDR3)，轻链可变区包括轻链CDR1(LCDR1)、轻链CDR2(LCDR2)和轻链CDR3(LCDR3)，其中：
    HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.2所示序列或与SEQ ID NO.2所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.6所示序列或与SEQ ID NO.6所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
    HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.17所示序列或与SEQ ID NO.17所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.32所示序列或与SEQ ID NO.32所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
    HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.17所示序列或与SEQ ID NO.17所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.6所示序列或与SEQ ID NO.6所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
    HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.2所示序列或与SEQ ID NO.2所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.32所示序列或与SEQ ID NO.32所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
    HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.21所示序列或与SEQ ID NO.21所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.32所示序列或与SEQ ID  NO.32所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
    HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.21所示序列或与SEQ ID NO.21所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.6所示序列或与SEQ ID NO.6所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
    HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.23所示序列或与SEQ ID NO.23所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.32所示序列或与SEQ ID NO.32所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
    HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.23所示序列或与SEQ ID NO.23所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.6所示序列或与SEQ ID NO.6所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
    HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.10所示序列或与SEQ ID NO.10所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.13所示序列或与SEQ ID NO.13所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；或
    HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.10所示序列或与SEQ ID NO.10所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.35所示序列或与SEQ ID NO.35所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；或
    HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.27所示序列或与SEQ ID NO.27所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.35所示序列或与SEQ ID NO.35所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；或
    HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.27所示序列或与SEQ ID NO.27所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.13所示序列或与SEQ ID NO.13所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；或
    HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.30所示序列或与SEQ ID NO.30所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.35所示序列或与SEQ ID NO.35所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；或
    HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.30所示序列或与SEQ ID NO.30所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR1的序列为SEQ ID NO.13所示序列或与SEQ ID NO.13所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；
    其中HCDR和LCDR序列根据Kabat定义。
  2. 如权利要求1所述的抗CD73抗体或其抗原结合部分，其中所述重链可变区具有SEQ ID NO.4、18、19、20、22、24、12、25、26、28、29或31所示氨基酸序列或与SEQ ID NO.4、18、19、20、22、24、12、25、26、28、29或31所示氨基酸序列具有至少80％、至少85％、至少90％、至少95％或至少99％同一性的氨基酸序列。
  3. 如权利要求1或2所述的抗CD73抗体或其抗原结合部分，其中所述轻链可变区具有SEQ ID NO.8、33、34、16、36、37或38所示氨基酸序列或与SEQ ID NO.8、33、34、16、36、37或38所示氨基酸序列具有至少80％、至少85％、至少90％、至少95％ 或至少99％同一性的氨基酸序列。
  4. 如权利要求1-3中任一项所述的抗CD73抗体或其抗原结合部分，其中
    重链可变区序列为SEQ ID NO.4所示序列，轻链可变区的序列为SEQ ID NO.8所示序列；或
    重链可变区序列为SEQ ID NO.18所示序列，轻链可变区的序列为SEQ ID NO.33所示序列；或
    重链可变区序列为SEQ ID NO.18所示序列，轻链可变区的序列为SEQ ID NO.34所示序列；或
    重链可变区序列为SEQ ID NO.19所示序列，轻链可变区的序列为SEQ ID NO.33所示序列；或
    重链可变区序列为SEQ ID NO.19所示序列，轻链可变区的序列为SEQ ID NO.34所示序列；或
    重链可变区序列为SEQ ID NO.20所示序列，轻链可变区的序列为SEQ ID NO.33所示序列；或
    重链可变区序列为SEQ ID NO.20所示序列，轻链可变区的序列为SEQ ID NO.34所示序列；或
    重链可变区序列为SEQ ID NO.22所示序列，轻链可变区的序列为SEQ ID NO.33所示序列；或
    重链可变区序列为SEQ ID NO.22所示序列，轻链可变区的序列为SEQ ID NO.34所示序列；或
    重链可变区序列为SEQ ID NO.24所示序列，轻链可变区的序列为SEQ ID NO.33所示序列；或
    重链可变区序列为SEQ ID NO.24所示序列，轻链可变区的序列为SEQ ID NO.34所示序列；或
    重链可变区序列为SEQ ID NO.12所示序列，轻链可变区的序列为SEQ ID NO.16所示序列；或
    重链可变区序列为SEQ ID NO.25所示序列，轻链可变区的序列为SEQ ID NO.36所示序列；或
    重链可变区序列为SEQ ID NO.25所示序列，轻链可变区的序列为SEQ ID NO.37所示序列；或
    重链可变区序列为SEQ ID NO.25所示序列，轻链可变区的序列为SEQ ID NO.38所示序列；或
    重链可变区序列为SEQ ID NO.26所示序列，轻链可变区的序列为SEQ ID NO.36所示序列；或
    重链可变区序列为SEQ ID NO.26所示序列，轻链可变区的序列为SEQ ID NO.37所示序列；或
    重链可变区序列为SEQ ID NO.26所示序列，轻链可变区的序列为SEQ ID NO.38所示序列；或
    重链可变区序列为SEQ ID NO.28所示序列，轻链可变区的序列为SEQ ID NO.36所示序列；或
    重链可变区序列为SEQ ID NO.28所示序列，轻链可变区的序列为SEQ ID NO.37所示序列；或
    重链可变区序列为SEQ ID NO.28所示序列，轻链可变区的序列为SEQ ID NO.38所示序列；或
    重链可变区序列为SEQ ID NO.29所示序列，轻链可变区的序列为SEQ ID NO.36所 示序列；或
    重链可变区序列为SEQ ID NO.29所示序列，轻链可变区的序列为SEQ ID NO.37所示序列；或
    重链可变区序列为SEQ ID NO.29所示序列，轻链可变区的序列为SEQ ID NO.38所示序列；或
    重链可变区序列为SEQ ID NO.31所示序列，轻链可变区的序列为SEQ ID NO.36所示序列；或
    重链可变区序列为SEQ ID NO.31所示序列，轻链可变区的序列为SEQ ID NO.37所示序列；或
    重链可变区序列为SEQ ID NO.31所示序列，轻链可变区的序列为SEQ ID NO.38所示序列。
  5. 抗CD73抗体或其抗原结合部分，所述抗CD73抗体或其抗原结合部分包括重链可变区和轻链可变区，其中：
    所述重链可变区具有SEQ ID NO.4、18、19、20、22、24、12、25、26、28、29或31所示氨基酸序列或与SEQ ID NO.4、18、19、20、22、24、12、25、26、28、29或31所示氨基酸序列具有至少80％、至少85％、至少90％、至少95％或至少99％同一性的氨基酸序列；和/或
    所述轻链可变区具有SEQ ID NO.8、33、34、16、36、37或38所示氨基酸序列或与SEQ ID NO.8、33、34、16、36、37或38所示氨基酸序列具有至少80％、至少85％、至少90％、至少95％或至少99％同一性的氨基酸序列。
  6. 抗CD73抗体或其抗原结合部分，所述抗CD73抗体或其抗原结合部分包括重链可变区，所述重链可变区包括重链CDR1(HCDR1)、重链CDR2(HCDR2)和重链CDR3(HCDR3)，其中：
    (1)HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.2所示序列或与SEQ ID NO.2所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性；或
    (2)HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.17所示序列或与SEQ ID NO.17所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性；或
    (3)HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.21所示序列或与SEQ ID NO.21所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性；或
    (4)HCDR1的序列为SEQ ID NO.1所示序列或与SEQ ID NO.1所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.23所示序列或与SEQ ID NO.23所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.3所示序列或与SEQ ID NO.3所示序列具有至少80％、85％、90％、95％或99％同一性；或
    (5)HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少 80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.10所示序列或与SEQ ID NO.10所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性；或
    (6)HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.27所示序列或与SEQ ID NO.27所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性；或
    (7)HCDR1的序列为SEQ ID NO.9所示序列或与SEQ ID NO.9所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR2的序列为SEQ ID NO.30所示序列或与SEQ ID NO.30所示序列具有至少80％、85％、90％、95％或99％同一性，HCDR3的序列为SEQ ID NO.11所示序列或与SEQ ID NO.11所示序列具有至少80％、85％、90％、95％或99％同一性；
    其中HCDR序列根据Kabat定义。
  7. 抗CD73抗体或其抗原结合部分，所述抗CD73抗体或其抗原结合部分包括轻链可变区，轻链可变区包括轻链CDR1(LCDR1)、轻链CDR2(LCDR2)和轻链CDR3(LCDR3)，其中：
    (1)LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.6所示序列或与SEQ ID NO.6所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
    (2)LCDR1的序列为SEQ ID NO.5所示序列或与SEQ ID NO.5所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.32所示序列或与SEQ ID NO.32所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.7所示序列或与SEQ ID NO.7所示序列具有至少80％、85％、90％、95％或99％同一性；或
    (3)LCDR1的序列为SEQ ID NO.13所示序列或与SEQ ID NO.13所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；或
    (4)LCDR1的序列为SEQ ID NO.35所示序列或与SEQ ID NO.35所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR2的序列为SEQ ID NO.14所示序列或与SEQ ID NO.14所示序列具有至少80％、85％、90％、95％或99％同一性，LCDR3的序列为SEQ ID NO.15所示序列或与SEQ ID NO.15所示序列具有至少80％、85％、90％、95％或99％同一性；
    其中LCDR序列根据Kabat定义。
  8. 如权利要求1-7中任一项所述的抗体或其抗原结合部分，其中：
    所述抗CD73抗体或其抗原结合部分结合人CD73；和/或
    所述抗CD73抗体或其抗原结合部分具有ADCC活性；和/或
    所述抗CD73抗体或其抗原结合部分具有CDC活性；和/或
    所述抗CD73抗体为全抗体、单链抗体(scFv)、scFv-Fc、双特异性抗体或多特异性抗 体；和/或
    所述抗CD73抗体的抗原结合部分为Fab、Fab’、Fv或F(ab’)2；和/或
    所述抗CD73抗体为鼠源抗体、嵌合抗体、人源化抗体或全人源抗体；和/或
    所述抗CD73抗体为单克隆抗体；和/或
    所述抗CD73抗体为IgG1、IgG2或IgG4同种型的；和/或
    所述抗CD73抗体包含κ亚型或者λ亚型的轻链恒定区；和/或
    所述抗CD73抗体包含人IgG1重链恒定区和人κ轻链恒定区；和/或
    所述抗CD73抗体包含野生型Fc区；和/或
    所述抗CD73抗体包含经过工程改造的Fc区，所述经过工程改造的Fc区使得所述抗体具有减弱的ADCC和/或CDC效应，例如，所述抗体具有人IgG1重链恒定区，并且含有L234、L235、P329、P331中一个或多个位点的突变，例如，L234A、L235A、P329A、P331S中的一种或多种突变，其中残基编号按照EU编号***。
  9. 如权利要求1-8中任一项所述的抗体或其抗原结合部分，其中所述抗体具有SEQ ID NO.20所示的重链可变区，SEQ ID NO.33所示的轻链可变区，SEQ ID NO.39所示的重链恒定区和SEQ ID NO.40所示的轻链恒定区。
  10. 如权利要求1-8中任一项所述的抗体或其抗原结合部分，其中所述抗体具有SEQ ID NO.31所示的重链可变区，SEQ ID NO.38所示的轻链可变区，SEQ ID NO.39所示的重链恒定区和SEQ ID NO.40所示的轻链恒定区。
  11. 如权利要求1-10中任一项所述的抗体或其抗原结合部分，其用于医疗疗法。
  12. 分离的核酸分子，其编码权利要求1-11中任一项所述的抗CD73抗体或其抗原结合部分。
  13. 载体，其含有权利要求12所述的核酸分子。
  14. 宿主细胞，其含有权利要求12所述的核酸分子或权利要求13所述的载体。
  15. 抗体药物偶联物，其包含与治疗剂偶联的权利要求1-11中任一项所述的抗CD73抗体或其抗原结合部分。
  16. 如权利要求15所述的抗体药物偶联物，其中所述治疗剂为抗肿瘤药物，例如，细胞毒性药物、免疫增强剂或放射性同位素。
  17. 药物组合物，其包含权利要求1-11中任一项所述的抗CD73抗体或其抗原结合部分或权利要求15或16所述的抗体药物偶联物，以及药学可接受的载体。
  18. 如权利要求17所述的药物组合物，其用于治疗个体中的肿瘤，特别是肿瘤细胞表面表达CD73(CD73+)的肿瘤。
  19. 如权利要求18所述的药物组合物，其中所述肿瘤为实体瘤。
  20. 如权利要求18所述的药物组合物，其中所述肿瘤为恶性肿瘤。
  21. 如权利要求18所述的药物组合物，其中所述肿瘤为癌症。
  22. 如权利要求18所述的药物组合物，其中所述肿瘤选自：卵巢癌、乳腺癌、黑色素瘤、结直肠癌、胃癌、肾癌、***癌、胰腺癌、食管癌、头颈癌、甲状腺癌、肺癌、膀胱癌、***、结肠癌、肝癌、腹膜癌、肝细胞癌、成胶质细胞瘤、尿路癌、直肠癌、子宫内膜癌、子宫癌、唾液腺癌、鳞状细胞癌、外阴癌、***癌、***癌、脑癌、淋巴瘤(例如B细胞淋巴瘤)、白血病，以及上述肿瘤的转移灶。
  23. 如权利要求18-22中任一项所述的药物组合物，其用于与免疫检查点抑制剂、A2AR拮抗剂或STAT-3抑制剂联合治疗所述肿瘤。
  24. 如权利要求23所述的药物组合物，其中所述免疫检查点抑制剂为抗PD-1抗体、抗PD-L1抗体或抗CTLA4抗体。
  25. 如权利要求23所述的药物组合物，其中所述免疫检查点抑制剂为抗PD-1抗体。
  26. 权利要求1-11任一项所述的抗CD73抗体或其抗原结合部分、权利要12所述所述的核酸分子、权利要求13所述的载体、权利要求14所述的宿主细胞或权利要求15或16所述的抗体药物偶联物在制备用于治疗个体中的肿瘤，特别是肿瘤细胞表面表达CD73(CD73+)的肿瘤的药物中的用途。
  27. 如权利要求26所述的用途，其中所述肿瘤为实体瘤。
  28. 如权利要求26所述的用途，其中所述肿瘤为恶性肿瘤。
  29. 如权利要求26所述的用途，其中所述肿瘤为癌症。
  30. 如权利要求26所述的用途，其中所述肿瘤选自：卵巢癌、乳腺癌、黑色素瘤、结直肠癌、胃癌、肾癌、***癌、胰腺癌、食管癌、头颈癌、甲状腺癌、肺癌、膀胱癌、***、结肠癌、肝癌、腹膜癌、肝细胞癌、成胶质细胞瘤、尿路癌、直肠癌、子宫内膜癌、子宫癌、唾液腺癌、鳞状细胞癌、外阴癌、***癌、***癌、脑癌、淋巴瘤(例如B细胞淋巴瘤)、白血病，以及上述肿瘤的转移灶。
  31. 如权利要求26-30中任一项所述的用途，其中所述药物用于与免疫检查点抑制剂、A2AR拮抗剂或STAT-3抑制剂联合治疗所述肿瘤。
  32. 如权利要求31所述的用途，其中所述免疫检查点抑制剂为抗PD-1抗体、抗PD-L1抗体或抗CTLA4抗体。
  33. 如权利要求31所述的用途，其中所述免疫检查点抑制剂为抗PD-1抗体。
  34. 治疗个体中的肿瘤，特别是肿瘤细胞表面表达CD73(CD73+)的肿瘤的方法，所述方法包括给予所述个体治疗有效量的权利要求1-11任一项所述的抗CD73抗体或其抗原结合部分、权利要求15或16所述的抗体药物偶联物或权利要求17所述的药物组合物。
  35. 如权利要求34所述的方法，其中所述肿瘤为实体瘤。
  36. 如权利要求34所述的方法，其中所述肿瘤为恶性肿瘤。
  37. 如权利要求34所述的方法，其中所述肿瘤为癌症。
  38. 如权利要求34所述的方法，其中所述肿瘤选自：卵巢癌、乳腺癌、黑色素瘤、结直肠癌、胃癌、肾癌、***癌、胰腺癌、食管癌、头颈癌、甲状腺癌、肺癌、膀胱癌、***、结肠癌、肝癌、腹膜癌、肝细胞癌、成胶质细胞瘤、尿路癌、直肠癌、子宫内膜癌、子宫癌、唾液腺癌、鳞状细胞癌、外阴癌、***癌、***癌、脑癌、淋巴瘤(例如B细胞淋巴瘤)、白血病，以及上述肿瘤的转移灶。
  39. 如权利要求34-38中任一项所述的方法，其中所述方法还包括将所述抗CD73抗体或其抗原结合部分、抗体药物偶联物或药物组合物与免疫检查点抑制剂、A2AR拮抗剂或STAT-3抑制剂联合给予所述个体，以治疗所述肿瘤。
  40. 如权利要求39所述的方法，其中所述免疫检查点抑制剂为抗PD-1抗体、抗PD-L1抗体或抗CTLA4抗体。
  41. 如权利要求39所述的方法，其中所述免疫检查点抑制剂为抗PD-1抗体。
  42. 缀合物，其包含权利要求1-11中任一项所述的抗CD73抗体或其抗原结合部分以及可检测标记物。
  43. 包含权利要求1-11中任一项所述的抗CD73抗体或其抗原结合部分的融合蛋白。
PCT/CN2023/072527 2022-01-25 2023-01-17 抗cd73抗体及应用 WO2023143227A1 (zh)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107001474A (zh) * 2014-11-21 2017-08-01 百时美施贵宝公司 抗cd73抗体及其用途
CN109476740A (zh) * 2016-03-04 2019-03-15 百时美施贵宝公司 利用抗cd73抗体的联合治疗
CN112166124A (zh) * 2018-05-19 2021-01-01 勃林格殷格翰国际有限公司 拮抗性cd73抗体

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107001474A (zh) * 2014-11-21 2017-08-01 百时美施贵宝公司 抗cd73抗体及其用途
CN109476740A (zh) * 2016-03-04 2019-03-15 百时美施贵宝公司 利用抗cd73抗体的联合治疗
CN112166124A (zh) * 2018-05-19 2021-01-01 勃林格殷格翰国际有限公司 拮抗性cd73抗体

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