Method of inhibition of vascular development using an antibody

Abstract

Tie1 and Tie2 are receptor tyrosine kinase proteins that include a transmembrane domain. Tie1 and Tie2 are present on endothelial cells. This disclosure describes agents, such as antibodies, that bind to Tie1, Tie2, and Ang, including ones that inhibit endothelial cell activity and angiogenesis. The agents can be used to treat angiogenesis-associated disorders.

1 - 28 . (canceled) 
     
     
         29 . An isolated human antibody, or antigen-binding fragment thereof, that binds to human Tie1 with an affinity K D  of less than 10 −8  M. 
     
     
         30 . The isolated human antibody, or antigen-binding fragment thereof, of  claim 29 , which binds to human Tie1 with an affinity K D  of less than 5×10 −9  M. 
     
     
         31 . The isolated human antibody, or antigen-binding fragment thereof, of  claim 29 , which binds to human Tie1 with an affinity K D  of less than 10 −9  M. 
     
     
         32 . An isolated human antibody, or antigen-binding fragment thereof, that binds to human Tie1 with a K on  rate of at least 6.19×10 3  Ms −1 . 
     
     
         33 . The isolated human antibody, or antigen-binding fragment thereof, of  claim 32 , which binds to human Tie1 with a K on  rate of at least 7.09×10 3  Ms −1 . 
     
     
         34 . An isolated human antibody, or antigen-binding fragment thereof, that binds to human Tie1 and has a K off  rate of 1.02×10 −3  s −1  or less. 
     
     
         35 . The isolated human antibody, or antigen-binding fragment thereof, of  claim 34 , which binds to human Tie1 and has a K off  rate of 3.67×10 −5  s −1  or less. 
     
     
         36 . The isolated human antibody, or antigen-binding fragment thereof, of any one of  claim 29 ,  32  or  34 , which is a recombinant antibody. 
     
     
         37 . The isolated human antibody, or antigen-binding fragment thereof, of any one of  claim 29 ,  32  or  34 , which is a Fab. 
     
     
         38 . The isolated human antibody, or antigen-binding fragment thereof, of any one of  claim 29 ,  32  or  34 , which is an IgG. 
     
     
         39 . An isolated human antibody or antigen-binding fragment thereof, which:
 induces Tie1 tyrosine phosphorylation;   inhibits tube formation by human umbilical endothelial cells (HUVECs) in vitro;   inhibits angiogenesis in a MATRIGEL™ in vivo assay; or   antagonizes the formation of a Tie1, Tie 2 and Ang heteromeric complex.   
     
     
         40 . The isolated human antibody, or antigen-binding fragment thereof, of  claim 39 , wherein the antibody, or antigen-binding fragment thereof,
 has a heavy chain CDR3 comprising the amino acid sequence of residues 99-109 of SEQ ID NO:114; and   has a light chain CDR3 comprising the amino acid sequence of residues 89-96 of SEQ ID NO:116.   
     
     
         41 . The isolated human antibody, or antigen-binding fragment thereof, of  claim 39 , wherein angiogenesis is inhibited by at least 70% in the MATRIGEL™ in vivo assay. 
     
     
         42 . The isolated human antibody, or antigen-binding fragment thereof, of  claim 39 , which has no deleterious effect on stem cells. 
     
     
         43 . The isolated human antibody, or antigen-binding fragment thereof, of  claim 39 , which does not induce platelet agglutination or aggregation. 
     
     
         44 . An isolated human antibody or antigen-binding fragment thereof, which dissociates from Tie1 with a K off  rate of 1.02×10 −3  s −1  or less and binds to the Tie1 ectodomain. 
     
     
         45 . An isolated human antibody or antigen-binding fragment thereof, with a light chain variable region (LCVR) having a CDR3 domain comprising the amino acid sequence of residues 89-96 of SEQ ID NO:116, and with a heavy chain variable region (HCVR) having a CDR3 domain comprising the amino acid sequence of residues 99-109 of SEQ ID NO:114. 
     
     
         46 . The isolated human antibody, or antigen-binding fragment thereof, of  claim 45 , wherein the LCVR further has a CDR2 domain comprising the amino acid sequence of residues 50-56 of SEQ ID NO:116 and the HCVR further has a CDR2 domain comprising the amino acid sequence of residues 50-66 of SEQ ID NO:114. 
     
     
         47 . The isolated human antibody, or antigen-binding fragment thereof, of  claim 45 , wherein the LCVR further has a CDR1 domain comprising the amino acid sequence of residues 24-34 of SEQ ID NO:116 and the HCVR further has a CDR1 domain comprising the amino acid sequence of residues 31-35 of SEQ ID NO:114. 
     
     
         48 . A pharmaceutical composition comprising a human antibody, or antigen binding portion thereof, and a pharmaceutically acceptable carrier, wherein the human antibody, or antigen-binding portion thereof, comprises a light chain CDR3 domain comprising the amino acid sequence of residues 89-96 of SEQ ID NO:116 and a heavy chain CDR3 domain comprising the amino acid sequence of residues 99-109 of SEQ ID NO:114. 
     
     
         49 . A pharmaceutical composition comprising the human antibody, or antigen-binding portion thereof, of  claim 29  and a pharmaceutically acceptable carrier. 
     
     
         50 . A method of inhibiting vascular development in a subject, the method comprising administering to a subject having or at risk for a disorder requiring inhibition of vascular development a human antibody, or antigen-binding fragment thereof, according to  claim 29 . 
     
     
         51 . The method of  claim 50 , wherein the human antibody, or antigen-binding fragment thereof, is administered in an amount effective to decrease vascular development in a subject. 
     
     
         52 . The method of  claim 50 , wherein the subject has a vascular-dependent cancer or tumor. 
     
     
         53 . The method of  claim 52 , wherein the tumor is a solid tumor. 
     
     
         54 . The method of  claim 50 , further comprising a second therapy that is an anti-cancer therapy. 
     
     
         55 . The method of  claim 54 , wherein the second therapy is chemotherapy. 
     
     
         56 . The method of  claim 54 , wherein the second therapy comprises administering bevacizumab. 
     
     
         57 . The method of  claim 54 , wherein the second therapy comprises administering an agent selected from the group consisting of 5-FU, leucovorin, avastin, cyclophosphamide, and/or irinotecan.