Humanized anti-ccr2 antibodies and methods of use therefor

Abstract

The present invention relates to a humanized antibody or functional fragment thereof which binds to a mammalian (e.g., human) CC-chemokine receptor 2 (CCR2) or a portion of the receptor and blocks binding of a ligand to the receptor. The invention further relates to a method of inhibiting the interaction of a cell bearing mammalian CCR2 with a ligand thereof, and to use of the antibodies and fragments in therapeutic, prophylactic and diagnostic methods.

1 . A humanized immunoglobulin or antigen-binding fragment thereof having binding specificity for CCR2, said immunoglobulin or fragment comprising an antigen binding region of nonhuman origin and at least a portion of an immunoglobulin of human origin. 
     
     
         2 .- 8 . (canceled) 
     
     
         9 . The humanized immunoglobulin or antigen-binding fragment thereof of  claim 1  comprising a heavy chain and a light chain, wherein said light chain comprises at least one complementarity determining region derived from an antibody of nonhuman origin which binds CCR2 and a framework region derived from a light chain of human origin, and wherein said heavy chain comprises at least one complementarity determining region derived from an antibody of nonhuman origin which binds CCR2 and a framework region derived from a heavy chain of human origin. 
     
     
         10 .- 14 . (canceled) 
     
     
         15 . A humanized immunoglobulin light chain or antigen-binding fragment thereof comprising CDR1, CDR2 and CDR3 of the light chain of murine 1D9 antibody and a human light chain framework region. 
     
     
         16 .- 17 . (canceled) 
     
     
         18 . An isolated nucleic acid molecule encoding the humanized immunoglobulin light chain or antigen-binding fragment thereof of  claim 15 . 
     
     
         19 . (canceled) 
     
     
         20 . A humanized immunoglobulin heavy chain or antigen-binding fragment thereof comprising CDR1, CDR2 and CDR3 of the heavy chain of murine 1D9 antibody and a human heavy chain framework region. 
     
     
         21 .- 22 . (canceled) 
     
     
         23 . An isolated nucleic acid molecule encoding the humanized immunoglobulin heavy chain or antigen-binding fragment thereof of  claim 20 . 
     
     
         24 . (canceled) 
     
     
         25 . The humanized immunoglobulin light chain or antigen-binding fragment thereof of  claim 15 , said light chain or antigen-binding fragment thereof having an amino acid sequence comprising at least a functional portion of the light chain variable region amino acid sequence of SEQ ID NO: 9. 
     
     
         26 . An isolated nucleic acid molecule comprising a nucleotide sequence encoding a humanized immunoglobulin light chain or antigen-binding fragment thereof of  claim 25 . 
     
     
         27 . (canceled) 
     
     
         28 . The humanized immunoglobulin heavy chain or antigen-binding fragment thereof of  claim 20 , said heavy chain or antigen-binding fragment thereof having an amino acid sequence comprising at least a functional portion of the heavy chain variable region amino acid sequence shown in SEQ ID NO: 10. 
     
     
         29 . An isolated nucleic acid molecule comprising a nucleotide sequence encoding the humanized immunoglobulin heavy chain or antigen-binding fragment thereof of  claim 28 . 
     
     
         30 . (canceled) 
     
     
         31 . An expression vector comprising a fused gene encoding a humanized immunoglobulin light chain, said gene comprising a nucleotide sequence of the nucleic acid molecule of  claim 18 . 
     
     
         32 . (canceled) 
     
     
         33 . A host cell comprising the expression vector of  claim 31 . 
     
     
         34 . An expression vector comprising a fused gene encoding a humanized immunoglobulin heavy chain, said gene comprising a nucleotide sequence of the nucleic acid molecule of  claim 23 . 
     
     
         35 . (canceled) 
     
     
         36 . A host cell comprising the expression vector of  claim 34 . 
     
     
         37 . A host cell comprising a first recombinant nucleic acid molecule encoding a humanized immunoglobulin light chain and a second recombinant nucleic acid molecule encoding a humanized immunoglobulin heavy chain, wherein said first nucleic acid molecule comprises the nucleic acid molecule of  claim 18 , and wherein said second nucleic acid molecule comprises a nucleotide sequence encoding a CDR derived from the heavy chain of murine antibody 1D9 and a framework region derived from a heavy chain of human origin. 
     
     
         38 . A method of preparing a humanized immunoglobulin comprising maintaining a host cell of  claim 37  under conditions appropriate for expression of a humanized immunoglobulin, whereby humanized immunoglobulin chains are expressed and a humanized immunoglobulin is produced. 
     
     
         39 .- 40 . (canceled) 
     
     
         41 . A method of inhibiting the interaction of a cell expressing CCR2 with a ligand of CCR2, comprising contacting said cell with an effective amount of a humanized immunoglobulin or antigen-binding fragment thereof of  claim 1 . 
     
     
         42 .- 45 . (canceled) 
     
     
         46 . A method of inhibiting HIV infection of a cell, comprising contacting a cell with an effective amount of a composition comprising a humanized immunoglobulin or antigen-binding fragment thereof of  claim 1 . 
     
     
         47 . A method of treating HIV in a patient comprising administering to the patient a composition comprising an effective amount of a humanized immunoglobulin or antigen-binding fragment thereof of  claim 1 . 
     
     
         48 . A method of inhibiting HIV infection in a patient, comprising administering to the patient a composition comprising an effective amount of a humanized immunoglobulin or antigen-binding fragment thereof of  claim 1 . 
     
     
         49 . A method of inhibiting a function associated with binding of a chemokine to mammalian CCR2, comprising contacting CCR2 with an effective amount of a humanized immunoglobulin or antigen-binding fragment thereof of  claim 1 , wherein said humanized immunoglobulin inhibits binding of said chemokine to mammalian CCR2 and inhibits one or more functions associated with binding of the chemokine to CCR2. 
     
     
         50 . (canceled) 
     
     
         51 . A method of inhibiting leukocyte trafficking in a patient, comprising administering to the patient a composition comprising an effective amount of a humanized immunoglobulin or antigen-binding fragment thereof of  claim 1  which binds to mammalian CCR2 or portion of CCR2 and inhibits binding of a ligand to the receptor. 
     
     
         52 .- 53 . (canceled) 
     
     
         54 . A method of treating a CCR2-mediated disorder in a patient, comprising administering to the patient an effective amount of a humanized immunoglobulin or antigen-binding fragment thereof of  claim 1  which binds to mammalian CCR2 or portion thereof. 
     
     
         55 . (canceled) 
     
     
         56 . A method of inhibiting restenosis in a patient, comprising administering to the patient an effective amount of a humanized immunoglobulin or antigen-binding fragment thereof of  claim 1  which binds to mammalian CCR2 or portion thereof. 
     
     
         57 .- 59 . (canceled) 
     
     
         60 . A pharmaceutical composition comprising a humanized immunoglobulin or antigen-binding fragment thereof of  claim 1  and a physiologically acceptable carrier. 
     
     
         61 . A humanized immunoglobulin light chain or antigen-binding fragment thereof having binding specificity for CCR2 comprising an amino acid sequence selected from the group consisting of:
 a) SEQ ID NO: 12;   b) SEQ ID NO: 13;   c) SEQ ID NO: 14;   d) SEQ ID NO: 15; and   e) SEQ ID NO: 107.   
     
     
         62 . A humanized immunoglobulin heavy chain or antigen-binding fragment thereof having binding specificity for CCR2 comprising an amino acid sequence selected from the group consisting of:
 a) SEQ ID NO: 17;   b) SEQ ID NO: 18;   c) SEQ ID NO: 19; and   d) SEQ ID NO: 20.   
     
     
         63 . The humanized immunoglobulin light chain or antigen-binding fragment thereof of  claim 15 , wherein the immunoglobulin light chain or antigen-binding fragment thereof is encoded by a nucleic acid molecule comprising SEQ ID NO: 98. 
     
     
         64 . The humanized immunoglobulin heavy chain or antigen-binding fragment thereof of  claim 18 , wherein the immunoglobulin heavy chain or antigen-binding fragment thereof is encoded by a nucleic acid molecule comprising SEQ ID NO: 97. 
     
     
         65 . A chimeric immunoglobulin or antigen-binding fragment thereof having binding specificity for CCR2 comprising a light chain variable region of nonhuman origin and a human constant region. 
     
     
         66 . A chimeric immunoglobulin or antigen-binding fragment thereof having binding specificity for CCR2 comprising a heavy chain variable region of nonhuman origin and a human constant region. 
     
     
         67 .- 71 . (canceled) 
     
     
         72 . A method of inhibiting narrowing of the lumen of a vessel in a mammal, comprising administering to said mammal an effective amount of a humanized immunoglobulin or antigen-binding fragment thereof of  claim 1  which binds to mammalian CCR2 or portion thereof. 
     
     
         73 . A method of inhibiting neointimal hyperplasia of a vessel in a mammal, comprising administering to said mammal an effective amount of a humanized immunoglobulin or antigen-binding fragment thereof of  claim 1  which binds to mammalian CCR2 or portion thereof. 
     
     
         74 .- 81 . (canceled) 
     
     
         82 . The humanized immunoglobulin or antigen-binding fragment thereof of  claim 9 , wherein said light chain comprises at least one complementarity determining region derived from murine monoclonal antibody 1D9 and a framework region derived from the light chain of human antibody HF-21/28, and wherein said heavy chain comprises at least one complementarity determining region derived from murine monoclonal antibody 1D9 and a framework region derived from the heavy chain of human antibody 4B4′CL. 
     
     
         83 .- 84 . (canceled) 
     
     
         85 . The humanized immunoglobulin or antigen-binding fragment thereof of  claim 1 , comprising a light chain and a heavy chain, wherein said light chain comprises a variable region comprising SEQ ID NO: 12. 
     
     
         86 . The humanized immunoglobulin or antigen-binding fragment thereof of  claim 1 , comprising a heavy chain and a light chain, wherein said heavy chain comprises a variable region comprising SEQ ID NO: 17. 
     
     
         87 .- 101 . (canceled)