Binding molecule specific for cd73 and use of binding molecule

Abstract

A binding molecule specific for CD73 and a use of the binding molecule. Specifically, provided are a separate antibody binding CD73 and inhibiting the activity of CD73 or an antigen binding part of the separate antibody, and a use of the separate antibody or the antigen binding part thereof in treatment of diseases; also provided are a nucleic acid molecule encoding the separate antibody or the antigen binding part thereof, an expression vector for expressing the separate antibody or the antigen binding part thereof, a host cell, and a preparation method.

1 - 71 . (canceled) 
     
     
         72 . An isolated antibody or antigen-binding fragment thereof comprising a heavy chain variable region that comprises HCDR1, HCDR2, HCDR3; and a light chain variable region that comprises LCDR1, LCDR2, LCDR3, wherein:
 (a) HCDR1 comprises the amino acid sequence of SEQ ID NO: 18;   (b) HCDR2 comprises the amino acid sequence of SEQ ID NO:19;   (c) HCDR3 comprises the amino acid sequence of SEQ ID NO:20;   (d) LCDR1 comprises the amino acid sequence of SEQ ID NO:23;   (e) LCDR2 comprises the amino acid sequence of SEQ ID NO:24; and   (f) LCDR3 comprises the amino acid sequence of SEQ ID NO: 25.   
     
     
         73 . The isolated antibody or antigen-binding fragment thereof of  claim 72 , which comprises:
 (i) a heavy chain variable region (VH) comprising an amino acid sequence which has at least 85% identity to the amino acid sequence of any one of SEQ ID NOs: 16, and 66-67, and comprises HCDR1 comprising SEQ ID NO: 18, HCDR2 comprising SEQ ID NO: 19, and HCDR3 comprising SEQ ID NO: 20; and   (ii) a light chain variable region (VL) comprising an amino acid sequence which has at least 85% identity to the amino acid sequence of any one of SEQ ID NO: 21, and 68-70, and comprises LCDR1 comprising SEQ ID NO: 23, LCDR2 comprising SEQ ID NO: 24, and LCDR3 comprising SEQ ID NO: 25.   
     
     
         74 . The isolated antibody or antigen-binding fragment thereof of  claim 72 , which comprises:
 1) a heavy chain variable region (VH) that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 16 and comprises HCDR1 comprising SEQ ID NO: 18, HCDR2 comprising SEQ ID NO: 19, and HCDR3 comprising SEQ ID NO: 20, and a light chain variable region (VL) that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO:21 and comprises LCDR1 comprising SEQ ID NO: 23, LCDR2 comprising SEQ ID NO: 24, and LCDR3 comprising SEQ ID NO: 25;   2) a heavy chain variable region (VH) that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 66 and comprises HCDR1 comprising SEQ ID NO: 18, HCDR2 comprising SEQ ID NO: 19, and HCDR3 comprising SEQ ID NO: 20, and a light chain variable region (VL) that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 68 and comprises LCDR1 comprising SEQ ID NO: 23, LCDR2 comprising SEQ ID NO: 24, and LCDR3 comprising SEQ ID NO: 25;   3) a heavy chain variable region (VH) that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 66 and comprises HCDR1 comprising SEQ ID NO: 18, HCDR2 comprising SEQ ID NO: 19, and HCDR3 comprising SEQ ID NO: 20, and a light chain variable region (VL) that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 69 and comprises LCDR1 comprising SEQ ID NO: 23, LCDR2 comprising SEQ ID NO: 24, and LCDR3 comprising SEQ ID NO: 25;   4) a heavy chain variable region (VH) that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 66 and comprises HCDR1 comprising SEQ ID NO: 18, HCDR2 comprising SEQ ID NO: 19, and HCDR3 comprising SEQ ID NO: 20, and a light chain variable region (VL) that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 70 and comprises LCDR1 comprising SEQ ID NO: 23, LCDR2 comprising SEQ ID NO: 24, and LCDR3 comprising SEQ ID NO: 25;   5) a heavy chain variable region (VH) that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 67 and comprises HCDR1 comprising SEQ ID NO: 18, HCDR2 comprising SEQ ID NO: 19, and HCDR3 comprising SEQ ID NO: 20, and a light chain variable region (VL) that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 68 and comprises LCDR1 comprising SEQ ID NO: 23, LCDR2 comprising SEQ ID NO: 24, and LCDR3 comprising SEQ ID NO: 25;   6) a heavy chain variable region (VH) that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 67 and comprises HCDR1 comprising SEQ ID NO: 18, HCDR2 comprising SEQ ID NO: 19, and HCDR3 comprising SEQ ID NO: 20, and a light chain variable region (VL) that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 69 and comprises LCDR1 comprising SEQ ID NO: 23, LCDR2 comprising SEQ ID NO: 24, and LCDR3 comprising SEQ ID NO: 25; or   7) a heavy chain variable region (VH) that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 67 and comprises HCDR1 comprising SEQ ID NO: 18, HCDR2 comprising SEQ ID NO: 19, and HCDR3 comprising SEQ ID NO: 20, and a light chain variable region (VL) that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 70 and comprises LCDR1 comprising SEQ ID NO: 23, LCDR2 comprising SEQ ID NO: 24, and LCDR3 comprising SEQ ID NO: 25.   
     
     
         75 . The isolated antibody or antigen-binding fragment thereof of  claim 72 , wherein the isolated antibody is an IgG. 
     
     
         76 . The isolated antibody or antigen-binding fragment thereof of  claim 72 , wherein the isolated antibody is an IgG1, IgG2 or IgG4. 
     
     
         77 . The isolated antibody or antigen-binding fragment thereof of  claim 72 , wherein the isolated antibody is a monoclonal antibody, a chimeric antibody, a humanized antibody, a human engineered antibody, a human antibody, Fv, a single chain antibody (scFv), Fab, Fab′, Fab′-SH or F(ab′) 2 . 
     
     
         78 . The isolated antibody or antigen-binding fragment thereof of  claim 72 , comprising a heavy chain and a light chain, wherein:
 (I) the heavy chain comprising an amino acid sequence which has at least 85% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 26, 27, 71 and 72, and comprises HCDR1 comprising SEQ ID NO: 18, HCDR2 comprising SEQ ID NO: 19, and HCDR3 comprising SEQ ID NO: 20; and   (II) the light chain comprising an amino acid sequence which has at least 85% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 28, 73, 74 and 75, and comprises LCDR1 comprising SEQ ID NO: 23, LCDR2 comprising SEQ ID NO: 24, and LCDR3 comprising SEQ ID NO: 25.   
     
     
         79 . The isolated antibody or antigen-binding fragment thereof of  claim 72 , which comprises:
 1) a heavy chain that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 26 and comprises HCDR1 comprising SEQ ID NO: 18, HCDR2 comprising SEQ ID NO: 19, and HCDR3 comprising SEQ ID NO: 20, and a light chain that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 28 and comprises LCDR1 comprising SEQ ID NO: 23, LCDR2 comprising SEQ ID NO: 24, and LCDR3 comprising SEQ ID NO: 25;   2) a heavy chain that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 27 and comprises HCDR1 comprising SEQ ID NO: 18, HCDR2 comprising SEQ ID NO: 19, and HCDR3 comprising SEQ ID NO: 20, and a light chain that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 28 and comprises LCDR1 comprising SEQ ID NO: 23, LCDR2 comprising SEQ ID NO: 24, and LCDR3 comprising SEQ ID NO: 25;   3) a heavy chain that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 71 and comprises HCDR1 comprising SEQ ID NO: 18, HCDR2 comprising SEQ ID NO: 19, and HCDR3 comprising SEQ ID NO: 20, and a light chain that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 73 and comprises LCDR1 comprising SEQ ID NO: 23, LCDR2 comprising SEQ ID NO: 24, and LCDR3 comprising SEQ ID NO: 25;   4) a heavy chain that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 71 and comprises HCDR1 comprising SEQ ID NO: 18, HCDR2 comprising SEQ ID NO: 19, and HCDR3 comprising SEQ ID NO: 20, and a light chain that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 74 and comprises LCDR1 comprising SEQ ID NO: 23, LCDR2 comprising SEQ ID NO: 24, and LCDR3 comprising SEQ ID NO: 25;   5) a heavy chain that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 71 and comprises HCDR1 comprising SEQ ID NO: 18, HCDR2 comprising SEQ ID NO: 19, and HCDR3 comprising SEQ ID NO: 20, and a light chain that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 75 and comprises LCDR1 comprising SEQ ID NO: 23, LCDR2 comprising SEQ ID NO: 24, and LCDR3 comprising SEQ ID NO: 25;   6) a heavy chain that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 72 and comprises HCDR1 comprising SEQ ID NO: 18, HCDR2 comprising SEQ ID NO: 19, and HCDR3 comprising SEQ ID NO: 20, and a light chain that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 73 and comprises LCDR1 comprising SEQ ID NO: 23, LCDR2 comprising SEQ ID NO: 24, and LCDR3 comprising SEQ ID NO: 25;   7) a heavy chain that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 72 and comprises HCDR1 comprising SEQ ID NO: 18, HCDR2 comprising SEQ ID NO: 19, and HCDR3 comprising SEQ ID NO: 20, and a light chain that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 74 and comprises LCDR1 comprising SEQ ID NO: 23, LCDR2 comprising SEQ ID NO: 24, and LCDR3 comprising SEQ ID NO: 25; or   8) a heavy chain that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 72 and comprises HCDR1 comprising SEQ ID NO: 18, HCDR2 comprising SEQ ID NO: 19, and HCDR3 comprising SEQ ID NO: 20, and a light chain that comprises an amino acid sequence which has at least 85% identity to the amino acid sequence of SEQ ID NO: 75 and comprises LCDR1 comprising SEQ ID NO: 23, LCDR2 comprising SEQ ID NO: 24, and LCDR3 comprising SEQ ID NO: 25.   
     
     
         80 . The isolated antibody or antigen-binding fragment thereof of  claim 79 , wherein the antibody comprises:
 1) a heavy chain consisting of SEQ ID NO: 26 and a light chain consisting of SEQ ID NO: 28;   2) a heavy chain consisting of SEQ ID NO: 27 and a light chain consisting of SEQ ID NO: 28;   3) a heavy chain consisting of SEQ ID NO: 71 and a light chain consisting of SEQ ID NO: 73;   4) a heavy chain consisting of SEQ ID NO: 71 and a light chain consisting of SEQ ID NO: 74;   5) a heavy chain consisting of SEQ ID NO: 71 and a light chain consisting of SEQ ID NO: 75;   6) a heavy chain consisting of SEQ ID NO: 72 and a light chain consisting of SEQ ID NO: 73;   7) a heavy chain consisting of SEQ ID NO: 72 and a light chain consisting of SEQ ID NO: 74; or   8) a heavy chain consisting of SEQ ID NO: 72 and a light chain consisting of SEQ ID NO: 75.   
     
     
         81 . The isolated antibody or antigen-binding fragment thereof of  claim 72 , which is an antagonist of CD73 or an antagonist of 5′-nucleotidase of CD73. 
     
     
         82 . The isolated antibody or antigen-binding fragment thereof of  claim 81 , wherein the CD73 is human CD73. 
     
     
         83 . A method of decreasing adenosine levels in a subject with tumor, improving a T cell response in a subject with tumor, stimulating an immune response in a subject, or inhibiting the growth of tumor cells in a subject, said method comprises administering a therapeutically effective amount of the isolated antibody or antigen-binding fragment thereof of  claim 72  to the subject. 
     
     
         84 . The method of  claim 83 , wherein the subject is a subject with tumor. 
     
     
         85 . The method of  claim 83 , wherein the subject has a tumor cell expressing CD73 and/or a tumor microenvironment containing CD73. 
     
     
         86 . A pharmaceutical composition comprising the isolated antibody or antigen-binding fragment thereof of  claim 72 , and a pharmaceutically acceptable excipient. 
     
     
         87 . A method of treating tumor comprising administering a therapeutically effective amount of the isolated antibody or antigen-binding fragment thereof of  claim 72  to a subject in need thereof. 
     
     
         88 . The method of  claim 87 , wherein the tumor is a solid tumor or hematological tumor. 
     
     
         89 . The method of  claim 87 , wherein the tumor is selected from bladder cancer, breast cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophageal cancer, gastrointestinal cancer, pancreatic cancer, colorectal cancer, colon cancer, renal cancer, head and neck cancer, lung cancer (small cell lung cancer or non-small cell lung cancer), stomach cancer, bone cancer, liver cancer, thyroid cancer, skin cancer, central nervous system tumor, lymphoma, leukemia, myeloma, sarcoma, and virus-associated cancer.