Mutants of interleukin-1 receptor antagonist and uses thereof

Abstract

The invention provides an isolated polypeptide comprising a variant amino acid sequence of SEQ ID NO: 1, or a fusion or derivative of said polypeptide, or a fusion of a said derivative thereof, wherein the polypeptide, fusion or derivative retains a biological activity of wild type IL-IRa. In one embodiment, the isolated polypeptide, fusion or derivative is or comprises a polypeptide variant of amino acid sequence SEQ ID NO: 1 comprising or consisting of substitutions at one or more of the following amino acid mutations of SEQ ID NO: 1: Q29K, P38Y, P38R, L42W, D47N, E52R, H54R, E90Y, Q129L, Q129N, M136N, M136D and Q149K. Also provided are pharmaceutical compositions of the above polypeptide, fusion or derivative, as well as uses of the same for treating a disease or condition capable of being treated by an agent which inhibits the function of IL-1 receptors.

1 . An isolated polypeptide comprising a variant of the amino acid sequence set forth SEQ ID NO: 1, wherein the polypeptide comprises at least 20 contiguous amino acids of SEQ ID NO: 1, wherein the polypeptide is between 140 and 180 amino acids in length, wherein the variant amino acid sequence of SEQ ID NO: 1 comprises an N84D amino acid substitution, wherein the polypeptide has an amino acid sequence which has at least 50% identity with SEQ ID NO: 1, and wherein the polypeptide comprises one or more amino acid substitutions at one or more of positions 1 to 11, 51 to 55, and/or 89 to 93 relative SEQ ID NO: 1, and wherein the polypeptide is capable of inhibiting IL-1β-induced NFκB activation in an in vitro reporter assay and/or inhibiting IL-1β-induced production of IL-6 in vitro and/or inhibiting collagen-induced arthritis in mice in vivo. 
     
     
         2 - 20 . (canceled) 
     
     
         21 . The isolated polypeptide of  claim 1 , wherein the polypeptide is a variant of amino acid sequence SEQ ID NO: 1 and further comprises substitutions at one or more of the following amino acid positions:
 Q11, V18, N19, Q29, P38, V40, L42, K45, D47, E52, P53, H54, E75, L78, Q79, A82, N84, E90, R92, K93, K96, D104, P107, T109, S110, Q129, M136, D138, V143, K145 and Q149.   
     
     
         22 - 49 . (canceled) 
     
     
         50 . A pharmaceutical formulation comprising a polypeptide, fusion or derivative according to  claim 1  in admixture with a pharmaceutically or veterinarily acceptable adjuvant, diluent or carrier. 
     
     
         51 . A pharmaceutical formulation according to  claim 50  suitable for parenteral administration to a human. 
     
     
         52 - 67 . (canceled) 
     
     
         68 . The isolated polypeptide according to  claim 1 , wherein the polypeptide comprises at least 30 contiguous amino acids of SEQ ID NO: 1. 
     
     
         69 . The isolated polypeptide according to  claim 1 , wherein the polypeptide is fused to another protein to produce a fusion protein. 
     
     
         70 . The isolated polypeptide according to  claim 69 , wherein
 (a) the C terminus of the polypeptide is fused to the Fc portion of a human IgG molecule, or   (b) the C terminus of the polypeptide is fused to human serum albumin, or   (c) the N terminus of the polypeptide is fused to human serum albumin.   
     
     
         71 . The pharmaceutical formulation of  claim 50 , wherein the formulation is suitable for ocular delivery.