Agr2 blocking antibody and use thereof

Abstract

Disclosed is an AGR2 blocking monoclonal antibody, and in particular, a humanized monoclonal antibody for blocking AGR2. Also disclosed is a pharmaceutical composition containing the antibody and a method for preparing the same, and a use of the antibody in blocking tumor growth and metastasis.

1 . An antibody or functional fragment thereof capable of specifically binding to AGR2 protein, which binds to essentially the same epitope of AGR2 protein as the murine anti-human AGR2 protein monoclonal antibody 18A4. 
     
     
         2 . The antibody or functional fragment thereof of  claim 1 , wherein the antibody is the murine anti-human monoclonal antibody 18A4, or a humanized or chimeric form thereof. 
     
     
         3 . The antibody or functional fragment thereof of  claim 1 , wherein said epitope is located within the protein disulfide isomerase active domain of AGR2. 
     
     
         4 . The antibody according to or functional fragment thereof of  claim 1 , wherein the AGR2 active domain to which the antibody binds is set forth in SEQ ID NO: 12. 
     
     
         5 . The antibody or functional fragment thereof of  claim 1 , comprising at least one sequence selected from the group consisting of
 the heavy chain CDR1 amino acid sequence of SEQ ID NO: 8, the heavy chain CDR2 amino acid sequence of SEQ ID NO: 9, the heavy chain CDR3 amino acid sequence of SEQ ID NO: 10, the light chain CDR1 amino acid sequence of SEQ ID NO: 11, the light chain CDR2 amino acid sequence of SEQ ID NO: 12, and the light chain CDR3 amino acid sequence of SEQ ID NO: 13.   
     
     
         6 . The antibody or functional fragment thereof of  claim 5  comprising the heavy chain CDR1 amino acid sequence as shown in DYNMD SEQ ID NO: 8), the heavy chain CDR2 amino acid sequence DINPNYDTTSYNQKFQG (SEQ ID NO: 9), the heavy chain CDR3 amino acid sequence SMMGYGSPMDY (SEQ ID NO: 10), the light chain CDR1 amino acid sequence RASKSVSTSGYSYMH (SEQ ID NO: 11), the light chain CDR2 amino acid sequence LASNLES (SEQ ID NO: 12), and the light chain CDR3 amino acid sequence QHIRELPRT (SEQ ID NO: 13). 
     
     
         7 . The antibody or functional fragment thereof of  claim 6 , comprising the heavy chain variable region amino acid sequence set forth in SEQ ID NO: 2, and the light chain variable region amino acid sequence set forth in SEQ ID NO: 1. 
     
     
         8 . The antibody or functional fragment thereof of  claim 6 , comprising the heavy chain variable region amino acid sequence set forth in SEQ ID NO: 4, and the light chain variable region amino acid sequence set forth in SEQ ID NO: 3. 
     
     
         9 . The antibody or functional fragment thereof of  claim 2 , wherein the humanized antibody is an intact IgG1 antibody. 
     
     
         10 . The antibody or functional fragment thereof of  claim 1 , wherein the functional fragment is selected from the group consisting of an Fab, an Fab′, an F(ab′) 2 , an Fv fragment, a linear antibody, a single chain antibody, and an Fab fragment. 
     
     
         11 . A pharmaceutical composition comprising the antibody or functional fragment thereof of  claim 1 , and a pharmaceutically acceptable carrier. 
     
     
         12 . An isolated nucleic acid or vector encoding the antibody or functional fragment thereof of  claim 1 . 
     
     
         13 . (canceled) 
     
     
         14 . A host cell comprising the vector of  claim 12 . 
     
     
         15 . A method for producing an antibody or functional fragment thereof comprising cultivating the host cell of  claim 14  so as to express the nucleic acid and produce the antibody or functional fragment thereof. 
     
     
         16 . The method of  claim 15 , further comprising recovering the antibody or functional fragment thereof from the culture of the host cell. 
     
     
         17 . A method for using the antibody or functional fragment thereof according to  claim 1  for the treatment of a disease associated with pathological angiogenesis in a mammal comprising the step of administering the antibody or functional fragment thereof to the mammal. 
     
     
         18 . The method of  claim 17 , wherein the disease is a cancer. 
     
     
         19 . The method of  claim 18 , wherein the cancer is selected from the group consisting of a breast cancer, an ovarian cancer, an osteosarcoma, a liver cancer, a pancreatic cancer, a prostate cancer, a colorectal cancer, a non-small cell lung cancer, a renal cancer, a head and neck cancer, a melanoma, and a multiple myeloma. 
     
     
         20 . The method of  claim 19 , wherein the treatment comprises the step of simultaneous or sequential administration of a second therapeutic agent with the antibody or functional fragment thereof. 
     
     
         21 . The method of  claim 20 , wherein the second therapeutic agent is selected from the group consisting of an anti-angiogenic agent, a chemotherapeutic agent, and a cytotoxic agent.