Polypeptides

Abstract

The present disclosure relates to a class of engineered polypeptides having a binding affinity for the neonatal Fc receptor (in the following referred to as FcRn), and provides an FcRn binding polypeptide comprising the sequence EX 2 X 3 X 4 AX 6 X 7 EIRWLPNL X 16 X 17 X 18 QRX 21 AFIX 25 X 26 LX 28 X 29 (SEQ ID NO:1075). The present disclosure also relates to the use of such an FcRn binding polypeptide as an agent for modifying pharmacokinetic and pharmacodynamic properties and as a therapeutic agent.

1 . An FcRn binding polypeptide, comprising an FcRn binding motif BM, which motif consists of the amino acid sequence 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 1075) 
                 
                     
                   EX 2  X 3  X 4  AX 6  X 7  EIR WLPNLX 16 X 17  X 18  QR 
                 
                     
                     
                 
                     
                   X 21  AFIX 25  X 26 LX 28  X 29   
                 
             
                
                
                
                
               
            
           
         
       
       wherein, independently from each other,
 X 2  is selected from A, D, E, F, H, I, K, L, N, Q, R, S, T, V, W and Y; 
 X 3  is selected from A, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W and Y; 
 X 4  is selected from A, D, E, F, G, H, I, K, L, N, Q, R, S, T, V, W and Y; 
 X 6  is selected from A, E, F, G, H, I, K, Q, R, S and V; 
 X 7  is selected from A, F, H, K, N, Q, R, S and V; 
 X 16  is selected from N and T; 
 X 17  is selected from F, W and Y; 
 X 18  is selected from A, D, E and N; 
 X 21  is selected from A, S, V and W; 
 X 25  is selected from D, E, G, H, I, K, L, N, Q, R, S, T, V, W and Y; 
 X 26  is selected from K and S, 
 X 28  is selected from A, D, E, F, H, I, K, L, N, Q, R, S, T, V, W and Y; and 
 X 29  is selected from D and R. 
 
     
     
         2 . The FcRn binding polypeptide according to  claim 1 , wherein the BM consists of an amino acid sequence selected from 
       
         
           
                 
                 
               
                     
                   i) 
                 
                     
                   (SEQ ID NO: 1076) 
                 
                     
                   EX 2  X 3  X 4  AX 6  HEIR WLPNLTX 17  X 18  QR 
                 
                     
                     
                 
                     
                   X 21  AFIX 25  KLX 28  D 
                 
             
                
                
                
                
                
               
            
           
         
       
       wherein, independently from each other,
 X 2  is selected from A, D, E, F, H, I, K, L, N, Q, R, S, T, V, W and Y; 
 X 3  is selected from A, D, E, G, H, K, L, M, N, Q, R, S, T, V and Y; 
 X 4  is selected from A, D, E, F, G, I, K, L, N, Q, R, S, T, V and Y; 
 X 6  is selected from A, G, K, R, S and V; 
 X 17  is selected from F, W and Y; 
 X 18  is selected from A, D, E and N; 
 X 21  is selected from A, S, V and W; 
 X 25  is selected from D, G, H, K, L, N, R, V and W; 
 X 28  is selected from A, D, E, H, K, L, N, Q, R, S, T, W and Y; and 
 ii) an amino acid sequence which has at least 96% identity to said sequence. 
 
     
     
         3 . The FcRn binding polypeptide according to  claim 1 , wherein sequence i) fulfills at least three of the six conditions I-VI:
 I. X 6  is selected from A, G, K and S,   II. X 7  is H;   III. X 17  is selected from F and Y;   IV. X 18  is D;   V. X 21  is selected from V and W;   VI. X 25  is selected from H and R.   
     
     
         4 . The FcRn binding polypeptide according to  claim 1 , wherein the sequence is selected from the group consisting of SEQ ID NO:1-353. 
     
     
         5 . The FcRn binding polypeptide according to  claim 1 , wherein said FcRn binding motif forms part of a three-helix bundle protein domain. 
     
     
         6 . The FcRn binding polypeptide according to  claim 1 , which comprises an amino acid sequence selected from: 
       
         
           
                 
                 
               
                     
                   iii) 
                 
                     
                   (SEQ ID NO: 1077) 
                 
                     
                   K-[ BM ]-DPSQS X a X b LLX c  EAKKL X d X e X f Q; 
                 
             
                
                
                
               
            
           
         
       
       wherein
 [BM] is an FcRn binding motif as defined herein, provided that X 29  is D; 
 X a  is selected from A and S, 
 X b  is selected from N and E; 
 X c  is selected from A, S and C; 
 X d  is selected from E, N and S, 
 X e  is selected from D, E and S, 
 X f  is selected from A and S, 
 
       and
 iv) an amino acid sequence which has at least 93% identity to a sequence defined by iii). 
 
     
     
         7 . The FcRn binding polypeptide according to  claim 6 , wherein sequence iii) is selected from the group consisting of SEQ ID NO:354-706. 
     
     
         8 . The FcRn binding polypeptide according to  claim 1 , which comprises an amino acid sequence selected from: 
       
         
           
                 
                 
               
                     
                   xi) 
                 
                     
                   (SEQ ID NO: 1092) 
                 
                     
                   AEAKYAK-[ BM ]-DPSQSSELLSEAKKLNDSQAPK; 
                 
             
                
                
                
               
            
           
         
       
       wherein [BM] is an FcRn binding motif as defined in  claim 1 ; and
 xii) an amino acid sequence which has at least 94% identity to the sequence defined in xi). 
 
     
     
         9 . The FcRn binding polypeptide according to  claim 8 , in which sequence xi) is selected from the group consisting of SEQ ID NO:1060-1062. 
     
     
         10 . The FcRn binding polypeptide according to  claim 1 , which comprises an amino acid sequence selected from: 
       
         
           
                 
                 
               
                     
                   xiii) 
                 
                     
                   (SEQ ID NO: 1095) 
                 
                     
                   VDAKYAK-[ BM ]-DPSQSSELLSEAKKLNDSQAPK, 
                 
             
                
                
                
               
            
           
         
         wherein [BM] is an FcRn binding motif as defined in  claim 1 ; and 
         xiv) an amino acid sequence which has at least 94% identity to the sequence defined in xiii). 
       
     
     
         11 . The FcRn binding polypeptide according to  claim 10 , in which sequence xiii) is selected from the group consisting of SEQ ID NO:707-1059. 
     
     
         12 . The FcRn binding polypeptide according to  claim 1 , which is capable of binding to FcRn at pH 6.0 such that the K D  value of the interaction is at most 1×10 −6  M. 
     
     
         13 . The FcRn binding polypeptide according to  claim 1 , wherein the K D  value of the interaction between FcRn binding polypeptide and FcRn at pH 7.4 is higher than the K D  value of said interaction at pH 6.0. 
     
     
         14 . The FcRn binding polypeptide according to  claim 1 , wherein the K D  value of the interaction between FcRn binding polypeptide and FcRn at pH 7.4 is at least 1×10 −8  M. 
     
     
         15 . The FcRn binding polypeptide according to  claim 1 , wherein the K D  value of the interaction between FcRn binding polypeptide and FcRn at pH 7.4 is at most 1×10 −6  M. 
     
     
         16 . A fusion protein or conjugate comprising
 a first moiety consisting of an FcRn binding polypeptide according to  claim 1 ; and   a second moiety consisting of a polypeptide having a desired biological activity.   
     
     
         17 . An FcRn binding polypeptide, fusion protein or conjugate according to  claim 1 , which inhibits binding of IgG to FcRn. 
     
     
         18 . A polynucleotide encoding a polypeptide according to  claim 1 . 
     
     
         19 . A composition comprising an FcRn binding polypeptide, fusion protein or conjugate according to  claim 1  and at least one pharmaceutically acceptable excipient or carrier. 
     
     
         20 . An FcRn binding polypeptide, fusion protein or conjugate according to  claim 1  for use as a medicament.