Humanized antibodies against west nile virus and therapeutic and prophylactic uses thereof

Abstract

The present invention relates to compositions comprising humanized antibodies or fragments thereof that immunospecifically bind to one or more antigens of a flavivirus, particularly of West Nile Virus (WNV) and methods for preventing, treating or ameliorating symptoms associated with a flavivirus, particularly of West Nile Virus (WNV) infection utilizing said compositions. In particular, the present invention relates to methods for preventing, treating or ameliorating symptoms associated with WNV infection, said methods comprising administering to a human subject an effective amount of one or more humanized antibodies or fragments thereof that immunospecifically bind to a WNV antigen. The present invention also relates to detectable or diagnostic compositions comprising humanized antibodies or fragments thereof that immunospecifically bind to a WNV. antigen and methods for detecting or diagnosing WNV infection utilizing said compositions.

1 - 16 . (canceled) 
     
     
         17 . A humanized antibody or an epitope binding fragment thereof comprising three VH complementary determining regions (CDRs) or three VL CDRs, wherein the antibody or epitope binding fragment thereof specifically binds a West Nile Virus (WNV) epitope defined by monoclonal antibody E16, E24, or E34, wherein said three VH or VL CDRs are from monoclonal antibody E16, E24, or E34 or differ by one amino acid substitution from a CDR from monoclonal antibody E16, E24, or E34. 
     
     
         18 . The humanized antibody of  claim 17  comprising a humanized variable heavy region comprising the amino acid sequence of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 37, or SEQ ID NO: 46. 
     
     
         19 . The humanized antibody of  claim 17  comprising a humanized variable light chain region comprising the amino acid sequence of SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 38, or SEQ ID NO: 47. 
     
     
         20 . The humanized antibody of  claim 17  comprising a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 27, a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 28, or SEQ ID NO: 39, and a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 3, SEQ ID NO: 29, or SEQ ID NO: 40. 
     
     
         21 . The humanized antibody of  claim 17  comprising a VL CDR1 having the amino acid sequence of SEQ ID NO: 11, a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 12, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 13 or SEQ ID NO: 34. 
     
     
         22 . The humanized antibody of  claim 17  comprising a framework region comprising at least one amino acid modification in heavy chain FR3 or light chain FR2. 
     
     
         23 . The humanized antibody of  claim 22 , wherein the at least one amino acid modification in heavy chain FR3 comprises a substitution at position 5, 6, 9, 11, 12, 19, 20, 25, 30, 38, 40, 43, 48, 66, 67, 69, 71, 75, 76, 79, 81, 82A, 83, 85, 87, 105, or 109. 
     
     
         24 . The humanized antibody of  claim 22 , wherein the at least one amino acid modification in light chain FR2 comprises a substitution at position 8, 9, 10, 11, 12, 13, 15, 17, 19, 20, 22, 43, 49, 63, 71, 78, 83, 85, or 100. 
     
     
         25 . The humanized antibody of  claim 17 , wherein the humanized antibody is an epitope-binding antibody fragment. 
     
     
         26 . The humanized antibody of  claim 25 , wherein the epitope-binding antibody fragment is a Fab, F(ab′) 2 , or scFv fragment. 
     
     
         27 . A pharmaceutical composition comprising (i) a therapeutically effective amount of the humanized antibody of  claim 17 ; and (ii) a pharmaceutically acceptable carrier. 
     
     
         28 . A method of treating a WNV infection in a patient, said method comprising administering to said patient a therapeutically effective amount of the humanized antibody of  claim 17 . 
     
     
         29 . The method of  claim 28  further comprising administering an anti-viral agent. 
     
     
         30 . The method of  claim 29 , wherein the anti-viral agent is selected from the group consisting of protease inhibitors, nucleoside reverse transcriptase inhibitors, non-nucleoside reverse transcriptase inhibitors, nucleoside analogs, and alpha-interferons. 
     
     
         31 . The method of  claim 29 , wherein the anti-viral agent is selected from the group consisting of zidovudine, acyclovir, gangcyclovir, vidarabine, idoxuridine, trifluridine, and ribavirin, foscarnet, amantadine, rimantadine, saquinavir, indinavir, amprenavir, lopinavir, ritonavir, adefovir, clevadine, entecavir, and pleconaril. 
     
     
         32 . The method of  claim 28 , wherein said patient is human. 
     
     
         33 . A method of diagnosis of WNV infection in a subject comprising:
 (a) contacting a biological sample from said subject with an effective amount of the humanized antibody of  claim 17 ; and   (b) detecting binding of said antibody,   
       wherein detection of said detectable marker above a background or standard level indicates that said subject has a WNV infection. 
     
     
         34 . The method of  claim 33 , wherein said detectable marker is a chemiluminescent, enzymatic, fluorescent, or radioactive label.