Il-17a/f heterologous polypeptides and therapeutic uses thereof

Abstract

The present invention is directed to a novel naturally occurring human cytokine that is comprised of a heterodimer of interleukin-17 and interleukin-17F designated herein as interleukin 17A/F (IL-17A/F). Also provided herein are vectors and host cells comprising those nucleic acid sequences, chimeric polypeptide molecules comprising the polypeptides of the present invention fused to heterologous polypeptide sequences, specific antibodies which bind to the polypeptides of the present invention and to methods for producing the polypeptides of the present invention. Further provided herein are methods for treating degenerative cartilaginous disorders and other inflammatory diseases.

1 . An isolated humanized antibody which binds to an IL 17A/F polypeptide comprising SEQ ID NO:3 and SEQ ID: with or without their associated signal peptides, wherein the antibody is an IgG 1  isotype, and wherein the antibody comprises an amino acid sequence comprising: 
       
         
           
                 
               
                   (SEQ ID NO: 81) 
                 
                   
                     LSCAASGFTXSDYXXXWVRQAPGKGLEWVATITXXGXNTYYXDSVKGRFT 
                   
                 
                     
                 
                     ISXDXXKNXXYLQMNSLRAEDTAVYYCAXXPXYYEGS . 
                 
             
                
                
                
                
               
            
           
         
       
     
     
         2 . The isolated antibody of  claim 1 , wherein said antibody is a monoclonal antibody, or a single-chain antibody. 
     
     
         3 . (canceled) 
     
     
         4 . The isolated antibody of claim which wherein the antibody is labeled and is immobilized on a solid support. 
     
     
         5 . The isolated antibody of  claim 1 , wherein said antibody is an antibody fragment. 
     
     
         6 .- 13 . (canceled) 
     
     
         14 . The isolated antibody of  claim 5 , wherein said antibody is an anti-IL-17A/F antagonist antibody. 
     
     
         15 . An isolated humanized antibody that binds to an IL-17A polypeptide comprising SEQ ID NO: 3 and to an IL-17F polypeptide comprising SEQ ID NO: 4, wherein the IL-17A and IL-17F polypeptides are in complex, wherein the antibody is an IgG 1  isotype, and wherein the antibody comprises an amino acid sequence comprising: 
       
         
           
                 
               
                   (SEQ ID NO: 81) 
                 
                   
                     LSCAASGFTXSDYXXXWVRQAPGKGLEWVATITXXGXNTYYXDSVKGRFT 
                   
                 
                     
                 
                     ISXDXXKNXXYLQMNSLRAEDTAVYYCAXXPXYYEGS . 
                 
             
                
                
                
                
               
            
           
         
       
     
     
         16 . An isolated humanized antibody that binds to an IL-17A polypeptide comprising SEQ ID NO: 3 or to an IL-17F polypeptide comprising SEQ ID NO: 4, wherein the IL-17A and IL-17F polypeptides are in a complex, wherein the antibody is an IgG 1  isotype, and wherein the antibody comprises an amino acid sequence comprising: 
       
         
           
                 
               
                   (SEQ ID NO: 81) 
                 
                   
                     LSCAASGFTXSDYXXXWVRQAPGKGLEWVATITXXGXNTYYXDSVKGRFT 
                   
                 
                     
                 
                     ISXDXXKNXXYLQMNSLRAEDTAVYYCAXXPXYYEGS . 
                 
             
                
                
                
                
               
            
           
         
       
     
     
         17 . An isolated humanized antibody that binds to an IL-17A polypeptide comprising SEQ ID NO: 3, wherein the IL-17A polypeptide is in a complex, wherein the antibody is an IgG 1  isotype, and wherein the antibody comprises an amino acid sequence comprising: 
       
         
           
                 
               
                   (SEQ ID NO: 81) 
                 
                   
                     LSCAASGFTXSDYXXXWVRQAPGKGLEWVATITXXGXNTYYXDSVKGRFT 
                   
                 
                     
                 
                     ISXDXXKNXXYLQMNSLRAEDTAVYYCAXXPXYYEGS . 
                 
             
                
                
                
                
               
            
           
         
       
     
     
         18 . An isolated humanized antibody that binds to an IL-17F polypeptide comprising SEQ ID NO: 4, wherein the IL-17A polypeptide is in a complex, wherein the antibody is an IgGi isotype, and wherein the antibody comprises an amino acid sequence comprising: 
       
         
           
                 
               
                   (SEQ ID NO: 81) 
                 
                   
                     LSCAASGFTXSDYXXXWVRQAPGKGLEWVATITXXGXNTYYXDSVKGRFT 
                   
                 
                     
                 
                     ISXDXXKNXXYLQMNSLRAEDTAVYYCAXXPXYYEGS . 
                 
             
                
                
                
                
               
            
           
         
       
     
     
         19 .- 22 . (canceled) 
     
     
         23 . An isolated humanized antibody which binds to an IL-17A/F polypeptide comprising SEQ ID NO:3 and SEQ ID NO:4 with or without their associated signal peptides, wherein the antibody is an IgG 4  isotype, and wherein the antibody comprises an amino acid sequence comprising: 
       
         
           
                 
               
                   (SEQ ID NO: 82) 
                 
                   SCXASGXXXTDYXIHWVRQAPGXGLEWXGVINPXXGXTDYXXXXKGRXTI 
                 
                     
                 
                   XADXSXXTAYXXXXSLRXEDTAVYYCARXXYFTGTGVY. 
                 
             
                
                
                
                
               
            
           
         
       
     
     
         24 . An isolated humanized antibody fragment which binds to an IL-17A/F polypeptide comprising SEQ ID NO:3 and SEQ ID NO:4 with or without their associated signal peptides, and wherein the antibody comprises an amino acid sequence comprising: 
       
         
           
                 
               
                   (SEQ ID NO: 83) 
                 
                   LSCAASGXTXXXXGWXRQAPGKXXEXVAXISXSGXDTYYADSVKGRFTIS 
                 
                     
                 
                   XDXXXXTXYLQMNSLXXEDTAVYYCAXRXGLYYVWDSNDY. 
                 
             
                
                
                
                
               
            
           
         
       
     
     
         25 . An isolated humanized antibody that binds to an IL-17A polypeptide comprising SEQ ID NO: 3 and to an IL-17F polypeptide comprising SEQ ID NO: 4, wherein the IL-17A and IL-17F polypeptides are in complex, wherein the antibody is an IgG 4  isotype, and wherein the antibody comprises an amino acid sequence comprising: 
       
         
           
                 
               
                   (SEQ ID NO: 82) 
                 
                   SCXASGXXXTDYXIHWVRQAPGXGLEWXGVINPXXGXTDYXXXXKGRXTI 
                 
                     
                 
                   XADXSXXTAYXXXXSLRXEDTAVYYCARXXYFTGTGVY. 
                 
             
                
                
                
                
               
            
           
         
       
     
     
         26 . An isolated humanized antibody fragment that binds to an IL-17A polypeptide comprising SEQ ID NO: 3 and to an IL-17F polypeptide comprising SEQ ID NO: 4, wherein the IL-17A and IL-17F polypeptides are in complex, and wherein the antibody comprises an amino acid sequence comprising: 
       
         
           
                 
               
                   (SEQ ID NO: 83) 
                 
                   LSCAASGXTXXXXGWXRQAPGKXXEXVAXISXSGXDTYYADSVKGRFTIS 
                 
                     
                 
                   XDXXXXTXYLQMNSLXXEDTAVYYCAXRXGLYYVWDSNDY. 
                 
             
                
                
                
                
               
            
           
         
       
     
     
         27 . An isolated humanized antibody that binds to an IL-17A polypeptide comprising SEQ ID NO: 3 or to an IL-17F polypeptide comprising SEQ ID NO: 4, wherein the IL-17A and IL-17F polypeptides are in a complex, wherein the antibody is an IgG 4  isotype, and wherein the antibody comprises an amino acid sequence comprising: 
       
         
           
                 
               
                   (SEQ ID NO: 82) 
                 
                   SCXASGXXXTDYXIHWVRQAPGXGLEWXGVINPXXGXTDYXXXXKGRXTI 
                 
                     
                 
                   XADXSXXTAYXXXXSLRXEDTAVYYCARXXYFTGTGVY. 
                 
             
                
                
                
                
               
            
           
         
       
     
     
         28 . An isolated humanized antibody that binds to an IL-17A polypeptide comprising SEQ ID NO: 3, wherein the IL-17A polypeptide is in a complex, wherein the antibody is an IgG4 isotype, and wherein the antibody comprises an amino acid sequence comprising: 
       
         
           
                 
               
                   (SEQ ID NO: 82) 
                 
                   SCXASGXXXTDYXIHWVRQAPGXGLEWXGVINPXXGXTDYXXXXKGRXTI 
                 
                     
                 
                   XADXSXXTAYXXXXSLRXEDTAVYYCARXXYFTGTGVY. 
                 
             
                
                
                
                
               
            
           
         
       
     
     
         29 . An isolated humanized antibody fragment that binds to an IL-17A polypeptide comprising SEQ ID NO: 3, wherein the IL-17A polypeptide is in a complex with an IL-17F polypeptide, and wherein the antibody comprises an amino acid sequence comprising: 
       
         
           
                 
               
                   (SEQ ID NO: 83) 
                 
                   LSCAASGXTXXXXGWXRQAPGKXXEXVAXISXSGXDTYYADSVKGRFTIS 
                 
                     
                 
                   XDXXXXTXYLQMNSLXXEDTAVYYCAXRXGLYYVWDSNDY. 
                 
             
                
                
                
                
               
            
           
         
       
     
     
         30 . An isolated humanized antibody that binds to an IL-17F polypeptide comprising SEQ ID NO: 4, wherein the IL-17A polypeptide is in a complex, wherein the antibody is an IgG4 isotype, and wherein the antibody comprises an amino acid sequence comprising: 
       
         
           
                 
               
                   (SEQ ID NO: 82) 
                 
                   SCXASGXXXTDYXIHWVRQAPGXGLEWXGVINPXXGXTDYXXXXKGRXTI 
                 
                     
                 
                   XADXSXXTAYXXXXSLRXEDTAVYYCARXXYFTGTGVY. 
                 
             
                
                
                
                
               
            
           
         
       
     
     
         31 . An isolated humanized antibody which binds to an IL-17A/F polypeptide comprising SEQ ID NO:3 and SEQ ID NO:4 with or without their associated signal peptides, wherein the antibody is an IgG 1  isotype, and wherein the antibody comprises three heavy chain CDR regions: a CDR1 region with an amino acid sequence comprising LSCAASGFTX 1 SDYX 2 X 3  (SEQ ID NO: 84), wherein X 1  and X 3  comprise a non-polar hydrophobic residue, and wherein X 2  comprises a polar hydrophilic residue; a CDR2 region with an amino acid sequence comprising APGKGLEWVATITX 4 X 5 G (SEQ ID NO: 85), wherein X 4  comprises a non-polar residue, and X 5  comprises a polar hydrophilic residue; and a CDR3 region with an amino acid sequence comprising YLQMNSLRAEDTAVYYCAX 6 X 7 PX 8 YYEGSX 9  (SEQ ID NO: 86), wherein X 6  and X 8  comprise a polar hydrophilic residue, and X 7  and X 9  comprise a non-polar hydrophobic residue. 
     
     
         32 . An isolated humanized antibody which binds to an IL-17A/F polypeptide comprising SEQ ID NO:3 and SEQ ID NO:4 with or without their associated signal peptides, wherein the antibody is an IgG4 isotype, and wherein the antibody comprises three heavy chain CDR regions: a CDR1 region with an amino acid sequence comprising ASGX a X b X c TDYX d  (SEQ ID NO: 87) wherein X a  comprises a non-polar aromatic residue, wherein X b  comprises a polar, hydrophilic neutral residue, wherein X c  comprises a non-polar hydrophobic residue, and wherein X d  comprises a polar hydrophilic residue; a CDR2 region with an amino acid sequence comprising GLEWX e GVINPX f X g GX h TDY (SEQ ID NO: 88), wherein X e , X f , and X g  comprise a non-polar hydrophobic residue, and wherein X h  comprises a polar, hydrophilic neutral residue; and a CDR3 region with an amino acid sequence comprising EDTAVYYCARX i X J YFTGTGVY (SEQ ID NO: 89), wherein X h  comprises a non-polar hydrophobic residue and X j  comprises a polar hydrophilic residue. 
     
     
         33 . An isolated humanized antibody fragment which binds to an IL-17A/F polypeptide comprising SEQ ID NO:3 and SEQ ID NO:4 with or without their associated signal peptides, and wherein the fragment comprises three heavy chain CDR regions: a CDR1 region with an amino acid sequence comprising TX 1′ X 2′ X 3′ X 4′ G, wherein X 1′ , X 3′  and X 4′  comprise a non-polar hydrophobic residue, and X 2′  comprises a polar hydrophilic residue; a CDR2 region with an amino acid sequence comprising EX 5′ VAX 6′ ISX 7′ SG (SEQ ID NO: 90), wherein X 5′  comprises an aromatic, non-polar hydrophobic residue, and wherein X 6′  and X 7′  comprise a non-polar hydrophobic residue; and a CDR3 region with an amino acid sequence comprising EDTAVYYCAX 8′ RX 9′ GLYYVWDSNDY (SEQ ID NO: 91), wherein X 8′  and X 9′  comprise a polar hydrophilic residue. 
     
     
         34 . The isolated antibody of  claim 1  or  15 , wherein the antibody binds to IL-17A/F polypeptide with a KD no greater than 1×10 −10  M. 
     
     
         35 . The isolated antibody of  claim 16  or  17 , wherein the antibody binds to IL-17A with a K D  no greater than 1×10 −10  M. 
     
     
         36 . The isolated antibody of  claim 18 , wherein the antibody binds to IL-17F with a K D  no greater than 1×10 −10  M. 
     
     
         37 . The isolated antibody of  claim 23  or  25 , wherein the antibody binds to IL-17A/F polypeptide with a K D  no greater than 1×10 −10  M. 
     
     
         38 . The isolated antibody of  claim 27  or  28 , wherein the antibody binds to IL-17A/F polypeptide with a K D  no greater than 1×10 −10  M.