Anti-AXL Antagonistic Antibodies

Abstract

Described are antibodies that specifically bind to the Axl protein and inhibit the interaction between Axl and the Axl-ligand, Gas6. Also disclosed are methods for the production and use of the anti-Axl antibodies.

1 . An antibody that binds to the epitope bound by the antibody obtainable from either:
 A) the hybridoma UT-10C9-B9; or   B) the hybridoma WR-10G5-E5.   
     
     
         2 . The antibody according to  claim 1 , that inhibits the binding of Axl to its ligand Gas6. 
     
     
         3 . The antibody according to either one of  claim 1  or  2  that downregulates Axl expression, inhibits Axl receptor signalling, and/or inhibits tumour growth. 
     
     
         4 . The antibody according to any one of  claims 1  to  3  that binds Axl and that comprises:
 A) an antibody VH domain comprising a VH CDR3 with the amino acid sequence of SEQ ID NO.7 and optionally one or more VH CDR's with an amino acid sequence selected from SEQ ID NO.6 and SEQ ID NO.5; and/or 
 an antibody VL domain comprising one or more VL CDR's with an amino acid sequence selected from SEQ ID NO.8, SEQ ID NO.9 and SEQ ID NO.10; 
 or 
 B) an antibody VH domain comprising a VH CDR3 with the amino acid sequence of SEQ ID NO.25 and optionally one or more VH CDR's with an amino acid sequence selected from SEQ ID NO.24 and SEQ ID NO.23; and/or 
 an antibody VL domain comprising one or more VL CDR's with an amino acid sequence selected from SEQ ID NO.26, SEQ ID NO.27 and SEQ ID NO.28. 
 
     
     
         5 . An antibody according to any one of  claims 1  to  4  comprising an antibody VH domain comprising either:
 A) the VH CDR's with the amino acid sequences of SEQ ID NO.5, SEQ ID NO.6 and SEQ ID NO.7, which antibody competes for binding to Axl with an Axl binding domain of an antibody comprising the 10C9 VH domain (SEQ ID NO. 3) and the 10C9 VL domain (SEQ ID NO. 4); or 
 B) the VH CDR's with the amino acid sequences of SEQ ID NO.23, SEQ ID NO.24 and SEQ ID NO.25, which antibody competes for binding to Axl with an Axl binding domain of an antibody comprising the 10G5 VH domain (SEQ ID NO. 21) and the 10G5 VL domain (SEQ ID NO. 22). 
 
     
     
         6 . An antibody according to any one of  claims 1  to  5  comprising either:
 A) the 10C9 VH domain (SEQ ID NO. 3); or 
 B) the 10G5 VH domain (SEQ ID NO. 21). 
 
     
     
         7 . An antibody according to  claim 6  comprising either:
 A) the 10C9 VL domain (SEQ ID NO. 4); or 
 B) the 10G5 VL domain (SEQ ID NO. 22). 
 
     
     
         8 . A variant of an antibody according to any one of  claims 1  to  7 , wherein the variant comprises one or more amino acid sequence alterations in one or more framework regions and/or one or more CDRs. 
     
     
         9 . An antibody according to any one of  claims 1  to  8 , wherein the antibody comprises:
 (i) all or a portion of an antibody heavy chain constant region and/or of an antibody light chain constant region; 
 (ii) a whole antibody; or 
 (iii) an antigen-binding antibody fragment. 
 
     
     
         10 . An antibody according to  claim 9 , wherein the whole antibody is an IgG antibody; or wherein the antigen-binding antibody fragment is a single domain antibody, Fv, scFv, dsFv, Fd, Fab, F(ab′)2, minibody, diabody, single-chain diabody, tandem scFv, TandAb, bi-body, tri-body, kappa(lambda)-body, BiTE, DVD-Ig, SIP, SMIP, or DART. 
     
     
         11 . An antibody according to any one of  claims 1  to  10  wherein the antibody is a humanized or a chimeric antibody. 
     
     
         12 . An antibody according to any one of  claims 1  to  11  wherein the Axl is human Axl. 
     
     
         13 . An antibody according to any one of  claims 1  to  12  that binds Axl with a K D  no greater than 6×10 −10  M. 
     
     
         14 . An antibody according to any one of  claims 1  to  13  that binds Axl with a k on  no lower than 8×10 5  M −1 s −1 . 
     
     
         15 . An antibody according to any one of  claims 1  to  14  that specifically binds primate Axl. 
     
     
         16 . An antibody according to any one of  claims 1  to  15  that:
 (i) binds murine Axl with a K D  greater than 10 −3  M; 
 (ii) binds human Mer with a K D  greater than 10 −3  M; and/or 
 (iii) binds human Tyro3 with a K D  greater than 10 −3  M. 
 
     
     
         17 . An antibody according to any one of  claims 1  to  16  that inhibits the binding of Axl to Gas6. 
     
     
         18 . An antibody according to any one of  claims 1  to  17  that down-regulates expression of the Axl receptor. 
     
     
         19 . An antibody according to any one of  claims 1  to  18  that increases the rate of Axl receptor internalization. 
     
     
         20 . An antibody according to any one of  claims 1  to  19 , wherein the antibody inhibits Axl receptor downstream signalling. 
     
     
         21 . An antibody according to any one of  claims 1  to  20  that increases the rate of cell death. 
     
     
         22 . An antibody according to any one of  claims 1  to  21  that inhibits tumour growth. 
     
     
         23 . An antibody according to any one of  claims 1  to  22  that is conjugated to a detectable label, enzyme, or toxin, optionally via a peptidyl bond or linker. 
     
     
         24 . An antibody obtainable from either:
 A) the hybridoma UT-10C9-B9; or   B) the hybridoma WR-10G5-E5.   
     
     
         25 . An isolated nucleic acid that comprises a nucleotide sequence encoding an antibody or antibody VH or VL domain of an antibody according to any one of  claims 1  to  23 . 
     
     
         26 . A host cell transformed with nucleic acid according to  claim 25 . 
     
     
         27 . A method of producing an antibody or antibody VH or VL domain, the method comprising culturing host cells according to  claim 26  under conditions for production of said antibody or antibody VH or VL domain. 
     
     
         28 . A composition comprising an antibody according to any one of  claims 1  to  23 , or an immunoconjugate thereof, in conjunction with a pharmaceutically acceptable excipient. 
     
     
         29 . The composition according to  claim 28 , further comprising an Immune Checkpoint Modulator, and/or an anti-tumour antibody specific for a target other than Axl. 
     
     
         30 . An antibody according to any one of  claims 1  to  23 , or the composition according to  claim 23 , for use in a method of treatment. 
     
     
         31 . An antibody or composition according to  claim 30  for use in a method of treating a proliferative disease. 
     
     
         32 . An antibody or composition according to  claim 31  where the proliferative disease is cancer. 
     
     
         33 . An antibody or composition according to  claim 32  where the cancer is metastatic cancer. 
     
     
         34 . Use of an antibody according to any one of  claims 1  to  23 , or the composition according to either one of  claim 28  or  claim 29 , in the manufacture of a medicament for treatment of a disease or disorder characterized by overexpression of Axl. 
     
     
         35 . A diagnostic kit comprising an antibody according to any one of  claims 1  to  24  and one or more reagents that allow determination of the binding of said member to metastatic cancer cells. 
     
     
         36 . A kit comprising an antibody according to any one of  claims 1  to  23 , or the composition according to either one of  claim 28  or  claim 29 .