Novel interleukin-17a-specific and interleukin-23-specific binding polypeptides and uses thereof

Abstract

The present invention relates to novel, specific-binding therapeutic and/or diagnostic polypeptides directed against the target of Swiss Prot Q16552 and novel, specific-binding therapeutic and/or diagnostic polypeptides directed against the target of Swiss Prot Q9NPF7. In addition, the present invention relates to novel, specific-binding therapeutic and/or diagnostic polypeptides directed against one or both of Swiss Prot Q16552 and Swiss Prot Q9NPF7. The invention also relates to nucleic acid molecules encoding such polypeptides and to methods for generation of such polypeptides and nucleic acid molecules. In addition, the invention is directed to compositions comprising the polypeptides, and therapeutic and/or diagnostic uses of these polypeptides.

1 - 187 . (canceled) 
     
     
         188 . A lipocalin mutein having binding specificity for IL-23p19,
 wherein the lipocalin mutein is a mutein of human tear lipocalin comprising, in comparison with the linear polypeptide sequence of mature human tear lipocalin (SEQ ID NO: 1), twenty or more mutated amino acid residues selected from the group consisting of: Arg 26→Trp; Glu 27→Val or Gln; Phe 28→Cys; Pro 29→Ala or Thr; Glu 30→Phe or Trp; Met 31→Ala or Asp; Asn 32→Asp; Leu 33→Asp or Glu; Glu 34→Pro; Met 55→Ile; Leu 56→Pro; Ile 57→Thr; Ser 58→Phe; Arg 60→Leu; Cys 61→Ala, Trp, or Tyr; Val 64→Glu; Cys 101→Ser; Glu 104→Arg or Ala; Leu 105→Cys; His 106→Tyr or Trp; Lys 108→Arg, His, or Gin; Arg 111→Pro; and Lys 114→Trp; and Cys 153→Ser, and wherein the lipocalin mutein has at least 85% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 12, and 13, or   wherein the lipocalin mutein is a mutein of human neutrophil gelatinase-associated lipocalin (hNGAL) comprising, in comparison with the linear polypeptide sequence of mature hNGAL (SEQ ID NO: 8), sixteen or more mutated amino acid residues selected from the group consisting of: Gln 28→His, Leu 36→Glu or Met, Ala 40→Leu or Tyr, Ile 41→Met or Leu, Gin 49→Thr, Arg, or Asp, Tyr 52→Ser, Thr or Glu, Ser 68→Trp or Arg, Leu 70→Glu or Asp, Arg 72→Ile or Gly, Lys 73→Phe, Met or Val, Asp 77→Lys or Trp, Trp 79→Gln or Phe, Arg 81→Gly or Thr, Asn 96→Gly or His, Tyr 100→Met, Leu 103→Met, Tyr 106→Phe, Lys 125→Tyr, Ser 127→Tyr, and Lys 134→Glu, and wherein the lipocalin mutein has at least 85% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 9, 10, and 15.   
     
     
         189 . The lipocalin mutein of  claim 188 , wherein the lipocalin mutein comprises one of the following sets of mutated amino acid residues in comparison with the linear polypeptide sequence of mature human tear lipocalin (SEQ ID NO: 1):
 (a) Arg 26→Trp; Glu 27→Gln; Pro 29→Thr; Glu 30→Trp; Met 31→Asp; Asn 32→Asp; Leu 33→Asp; Glu 34→Pro; Leu 56→Pro; Ser 58→Phe; Arg 60→Leu; Val 64→Glu; Glu 104→Ala; His 106→Tyr; and Lys 108→Gln;   (b) Arg 26→Trp; Glu 27→Gln; Pro 29→Thr; Glu 30→Trp; Met 31→Ala; Asn 32→Asp; Leu 33→Glu; Glu 34→Pro; Met 55→Ile; Leu 56→Pro; Ile 57→Thr; Ser 58→Phe; Arg 60→Leu; Val 64→Glu; Glu 104→Arg; His 106→Trp; and Lys 108→Arg; or   (c) Arg 26→Trp; Glu 27→Val; Pro 29→Ala; Glu 30→Phe; Met 31→Asp; Asn 32→Asp; Leu 33→Asp; Glu 34→Pro; Met 55→Ile; Leu 56→Pro; Ile 57→Thr; Ser 58→Phe; Arg 60→Leu; Val 64→Glu; Glu 104→Ala; and Lys 108→His.   
     
     
         190 . The lipocalin mutein of  claim 188 , wherein the lipocalin mutein comprises one of the following sets of mutated amino acid residues in comparison with the linear polypeptide sequence of mature hNGAL (SEQ ID NO: 8):
 (a) Leu 36→Met, Ala 40→Leu, Ile 41→Met, Gln 49→Thr, Tyr 52→Ser, Ser 68→Trp, Leu 70→Asp, Lys 73→Phe, Asp 77→Lys, Trp 79→Gln, Arg 81→Gly, Cys 87→Ser, Asn 96→Gly, Tyr 100→Met, Leu 103→Met, Tyr 106→Phe, Lys 125→Tyr, Ser 127→Tyr, and Lys 134→Glu;   (b) Ala 40→Thr, Gln 49→Asp, Tyr 52→Glu, Leu 70→Asp, Arg 72→Ile, Lys 73→Met, Asp 77→Trp, Trp 79→Phe, Arg 81→Thr, Cys 87→Ser, Asn 96→His, Tyr 100→Met, Leu 103→Met, Tyr 106→Phe, Lys 125→Tyr, Ser 127→Tyr, Tyr 132→Phe, and Lys 134→Glu; or   (c) Leu 36→Glu, Ala 40→Leu, Ile 41→Leu, Gln 49→Arg, Tyr 52→Thr, Ser 68→Arg, Leu 70→Glu, Arg 72→Gly, Lys 73→Val, Cys 76→Tyr, Asp 77→Lys, Trp 79→Gln, Arg 81→Gly, Cys 87→Ser, Asn 96→Gly, Tyr 100→Met, Leu 103→Met, Tyr 106→Phe, Lys 125→Tyr, Ser 127→Tyr, Lys 134→Glu, Cys 175→Ala.   
     
     
         191 . The lipocalin mutein of  claim 188 , wherein the lipocalin mutein has at least 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 9, 10, 12, 13, and 15. 
     
     
         192 . The lipocalin mutein of  claim 188 , wherein the lipocalin mutein comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 9, 10, 12, 13, and 15. 
     
     
         193 . The lipocalin mutein of  claim 188 , wherein the lipocalin mutein is capable of binding IL-23p19 with a K D  value of about 140 nM or lower. 
     
     
         194 . The lipocalin mutein of  claim 188 , wherein the lipocalin mutein is capable of binding IL-23p19 with a K D  value of about 12 nM or lower. 
     
     
         195 . The lipocalin mutein of  claim 188 , wherein the lipocalin mutein is capable of inhibiting the binding of IL-23 to its receptor with an IC 50  value of about 120 nM or lower. 
     
     
         196 . The lipocalin mutein of  claim 188 , wherein the lipocalin mutein is capable of inhibiting the binding of IL-23 to its receptor with an IC 50  value of about 1 nM or lower. 
     
     
         197 . The lipocalin mutein of  claim 188 , wherein the lipocalin mutein is capable of blocking biological activity of IL-23. 
     
     
         198 . The lipocalin mutein of  claim 188 , wherein the lipocalin mutein is capable of blocking biological activity of IL-23 in a cell-based proliferation assay with an IC 50  value lower than the IC 50  value of ustekinumab. 
     
     
         199 . The lipocalin mutein of  claim 188 , wherein the lipocalin mutein is crossreactive with human and cynomolgus monkey IL-23. 
     
     
         200 . The lipocalin mutein of  claim 188 , wherein the lipocalin mutein is fused at its N-terminus and/or its C-terminus to a second lipocalin mutein or a moiety which is a protein, a protein domain, or a peptide. 
     
     
         201 . The lipocalin mutein of  claim 200 , wherein the second lipocalin mutein is is a lipocalin mutein having at least 85% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 9, 10, 12, 13, and 15, or a fragment or variant thereof. 
     
     
         202 . The lipocalin mutein of  claim 200 , wherein the second lipocalin mutein has a binding specificity for IL-17A. 
     
     
         203 . The lipocalin mutein of  claim 188 , wherein the lipocalin mutein is conjugated to a moiety that extends the serum half-life of the mutein, wherein the moiety that extends the serum half-life is selected from the group consisting of a polyalkylene glycol molecule, a polyethylene glycol molecule, hydroxyethyl starch, a Fc part of an immunoglobulin, a CH3 domain of an immunoglobulin, a CH4 domain of an immunoglobulin, an albumin binding peptide, and an albumin binding protein. 
     
     
         204 . A method of binding of IL-23p19 in a subject, comprising administering an effective amount of one or more lipocalin muteins of  claim 188 . 
     
     
         205 . A method for inhibiting the binding of IL-23 to its receptor in a subject, comprising administering an effective amount of one or more lipocalin muteins of  claim 188 . 
     
     
         206 . A nucleic acid molecule comprising a nucleotide sequence encoding the lipocalin mutein of  claim 188 . 
     
     
         207 . A method of producing the lipocalin mutein of  claim 188 , wherein the lipocalin mutein is produced starting from the nucleic acid encoding the lipocalin mutein.