Conjugate of an antibody against ccr5 and an antifusogenic peptide

Abstract

The current invention is related to a conjugate comprising one or more antifusogenic peptides and an anti-CCR5 antibody (mAb CCR5) characterized in that one to eight antifusogenic peptides are each conjugated to one terminus of the heavy and/or light chains of said anti-CCR5 antibody and to the pharmaceutical use of said conjugate.

1 . A conjugate comprising an anti-CCR5 antibody (mAb CCR5) having termini comprising an N-terminus and a C-terminus; and
 an antifusogenic peptide, conjugated to said N-terminus or said C-terminus.   
     
     
         2 . The conjugate of  claim 1 , comprising two antifusogenic peptides, conjugated to said anti-CCR5 antibody termini. 
     
     
         3 . The conjugate according to  claim 1 , wherein said antifusogenic peptides are linear peptides that comprise an amino acid sequence of from 5 to 100 amino acids. 
     
     
         4 . The conjugate of  claim 1 , wherein said antifusogenic peptide is conjugated to said N-terminus or C-terminus through a peptide linker. 
     
     
         5 . The conjugate of  claim 1 , wherein said mAb CCR5 comprises two heavy chains and two light chains. 
     
     
         6 . The conjugate of  claim 5 , characterized by the general formula mAb CCR5-[linker] m -[antifusogenic peptide] n , wherein m is independently for each antifusogenic peptide either 0 or 1, and n is an integer of from 1 to 8. 
     
     
         7 . The conjugate of  claim 6 , comprising a conjugate of heavy and/or light chain of mAb CCR5 and an antifusogenic peptide(s) (“chain conjugate”), selected from the group consisting of:
   [antifusogenic peptide]-[linker] m -[heavy chain];  (1)
 
   [heavy chain]-[linker] m -[antifusogenic peptide];  (2)
 
   [antifusogenic peptide]-[linker] m -[heavy chain]-[antifusogenic peptide];  (3)
 
   [antifusogenic peptide]-[linker] m -[light chain];  (4)
 
   [light chain]-[linker] m -[antifusogenic peptide];  (5)
 
   [antifusogenic peptide]-[linker] m -[light chain]-[antifusogenic peptide];  (6)
 
   [antifusogenic peptide]-[linker] m -[heavy chain]-[linker] m -[antifusogenic peptide];  (7)
 
   and 
   [antifusogenic peptide]-[linker] m -[light chain]-[linker] m -[antifusogenic peptide],  (8)
 
 
       wherein each linker can be the same or different, wherein each m is independently an integer of 1 or 0. 
     
     
         8 . The conjugate of  claim 7 , which comprises a chain conjugate (2), (3), (4), or (7). 
     
     
         9 . The conjugate of  claim 8 , which comprises:
   2×[mAb CCR5 light chain] and 2×(2),
     2×[mAb CCR5 light chain] and 2×(3),
     2×[mAb CCR5 heavy chain] and 2×(4), or
     2×[mAb CCR5 light chain] and 2×(7).
   
     
     
         10 . The conjugate of  claim 1 , wherein said antifusogenic peptide has a sequence selected from the group SEQ ID NO:29 to 35. 
     
     
         11 . The conjugate according of  claim 1 , wherein said anti-CCR5 antibody comprises a variable heavy chain domain consisting of an immunoglobulin framework and a CDR3 region selected from the group consisting of the heavy chain CDR3 sequences SEQ ID NO:16 and SEQ ID NO:17. 
     
     
         12 . The conjugate of  claim 1 , wherein said anti-CCR5 antibody comprises a variable heavy chain domain consisting of an immunoglobulin framework and a CDR3 region selected from the group consisting of heavy chain CDR3 sequences SEQ ID NOS:16 and 17; a CDR2 region selected from the group consisting of heavy chain CDR2 sequences SEQ ID NOS:13, 14, and 15; and a CDR1 region selected from the group consisting of heavy chain CDR1 sequences SEQ ID NOS:9, 10, 11, and 12. 
     
     
         13 . The conjugate of  claim 1 , wherein said anti-CCR5 antibody comprises a heavy chain variable domain selected from the group consisting of SEQ ID NOS:1, 3, 5, and 7. 
     
     
         14 . The conjugate of  claim 1 , wherein said anti-CCR5 antibody comprises a variable light chain domain consisting of an immunoglobulin framework and a CDR1 region selected from SEQ ID NOS:18, 19, and 20; a CDR2 region selected from SEQ ID NOS:21, 22, and 23; and a CDR3 region selected from SEQ ID NOS:24 and 25. 
     
     
         15 . The conjugate of  claim 12 , wherein said anti-CCR5 antibody comprises as heavy chain CDRs the CDRs of SEQ ID NO: 1 and as light chain CDRs the CDRs of SEQ ID NO: 2, as heavy chain CDRs the CDRs of SEQ ID NO: 3 and as light chain CDRs the CDRs of SEQ ID NO: 4, as heavy chain CDRs the CDRs of SEQ ID NO: 5 and as light chain CDRs the CDRs of SEQ ID NO: 6, or as heavy chain CDRs the CDRs of SEQ ID NO: 7 and as light chain CDRs the CDRs of SEQ ID NO: 8. 
     
     
         16 . The conjugate of  claim 15 , wherein said anti-CCR5 antibody comprises a variable heavy chain domain and a variable light chain domain independently selected from the group consisting of
 a) the heavy chain (V H ) variable domain defined by amino acid sequence SEQ ID NO:1 and the light chain (V L ) variable domain defined by SEQ ID NO:2;   b) the heavy chain variable domain defined by amino acid sequence SEQ ID NO:3 and the light chain variable domain defined by SEQ ID NO:4;   c) the heavy chain variable domain defined by amino acid sequence SEQ ID NO:5 and the light chain variable domain defined by SEQ ID NO:6;   d) the heavy chain variable domain defined by amino acid sequence SEQ ID NO:7 and the light chain variable domain defined by SEQ ID NO:8.   
     
     
         17 . The conjugate of  claim 1 , wherein said conjugate comprises:
 said anti-CCR5 antibody, selected from the group consisting of
 the heavy chain (V H ) variable domain defined by amino acid sequence SEQ ID NO:1 and the light chain (V L ) variable domain defined by SEQ ID NO:2; 
 the heavy chain variable domain defined by amino acid sequence SEQ ID NO:3 and the light chain variable domain defined by amino acid SEQ ID NO:4; 
 the heavy chain variable domain defined by amino acid sequence SEQ ID NO:5 and the light chain variable domain defined by amino acid SEQ ID NO:6; and 
 the heavy chain variable domain defined by amino acid sequence SEQ ID NO:7 and the light chain variable domain defined by amino acid SEQ ID NO:8; 
   a linker selected from the group consisting of amino acids glycine (G) and asparagine (N), the tripeptide GST, and SEQ ID NO: 36-62; and   an antifusogenic peptide having a sequence selected from the group consisting of SEQ ID NO:29 to 35.   
     
     
         18 . The conjugate of  claim 1 , wherein said antifusogenic peptide has a sequence selected from the group consisting of SEQ ID NOS:29 to 35. 
     
     
         19 . The conjugate of  claim 17 , wherein said conjugate comprises two light chain variable domains of SEQ ID NO:2, two conjugates of type (2) each comprising a heavy chain variable domain of SEQ ID NO:1, a linker of SEQ ID NO:40 and an antifusogenic peptide of SEQ ID NO:33, in comprising two light chain variable domains of SEQ ID NO:4, two conjugates of type (2) each comprising a heavy chain variable domain of SEQ ID NO:3, a linker of SEQ ID NO:40 and an antifusogenic peptide of SEQ ID NO:33, in comprising two light chain variable domains of SEQ ID NO:6, two conjugates of type (2) each comprising a heavy chain variable domain of SEQ ID NO:5, a linker of SEQ ID NO:40 and an antifusogenic peptide of SEQ ID NO:33, or in comprising two light chain variable domains of SEQ ID NO:8, two conjugates of type (2) each comprising a heavy chain variable domain of SEQ ID NO:7, a linker of SEQ ID NO:40 and an antifusogenic peptide of SEQ ID NO:33. 
     
     
         20 . The conjugate of  claim 1 , wherein said anti-CCR5 antibody is of IgG4 subclass, or is of IgG1 or IgG2 subclass with a mutation in amino acid 5228, L234, L235, and/or D265, and/or contains the PVA236 mutation 
     
     
         21 . A conjugate according to  claim 20 , wherein said anti-CCR5 antibody of IgG4 subclass has the mutation S228P and said anti-CCR5 antibody of IgG1 subclass has the mutations L234A and L235A. 
     
     
         22 . A method producing a conjugate of  claim 1 , wherein said method comprises:
 a) cultivating a cell containing nucleic acid molecules encoding a conjugate according to  claim 1  under conditions suitable for the expression of the conjugate; and   b) recovering the conjugate from the cell or the cell culture supernatant.   
     
     
         23 . A pharmaceutical composition, comprising an effective amount of a conjugate of  claim 1 , together with a pharmaceutically acceptable excipient or carrier. 
     
     
         24 . A method for treating a viral infection, comprising:
 administering an effective amount of a conjugate of  claim 1  to a subject in need thereof.   
     
     
         25 . The method of  claim 24 , wherein the viral infection is HIV infection.