Glycopegylated Factor IX

Abstract

Conjugates between Factor IX and PEG moieties. are disclosed in the present application. The conjugates are linked via a glycosyl linking group interposed between and covalently attached to the peptide and the modifying group. Conjugates are formed from glycosylated peptides by the action of a glycosyltransferase. The glycosyltransferase ligates a modified sugar moiety onto a glycosyl residue on the peptide. Also provided are methods for preparing the conjugates, methods for treating various disease conditions with the conjugates, and pharmaceutical formulations including the conjugates.

1 .- 5 . (canceled) 
     
     
         6 . A PEGylated Factor IX conjugate comprising a Factor IX peptide and a poly(ethylene glycol) moiety, wherein the poly(ethylene glycol) moiety is attached to the Factor IX peptide via a glycosyl linking group of a glycosyl residue as set forth by the formula: 
       
         
           
           
               
               
           
         
         wherein 
         R 2  is COOH; 
         R 3  is H; 
         R 4  is OH; 
         D is OH; and 
         G is R 1 -L- 
         wherein 
         L is a substituted heteroalkyl linker comprising C(O)CH 2 NH; 
         R 1  is the poly(ethylene glycol) moiety, wherein the poly(ethylene glycol) moiety comprises a branched poly(ethylene glycol) having a core glycerol moiety, and wherein the branched poly(ethylene glycol) has a molecular weight of about 40 kDa. 
       
     
     
         7 . The conjugate according to  claim 6 , wherein the Factor IX peptide comprises an amino acid sequence set forth as SEQ ID NO:1. 
     
     
         8 . The conjugate according to  claim 7 , wherein the poly(ethylene glycol) moiety is conjugated to the Factor IX peptide at Asn 157 or Asn 167. 
     
     
         9 . A pharmaceutical formulation comprising a PEGylated Factor IX conjugate according to  claim 6  and a pharmaceutically acceptable carrier. 
     
     
         10 . A method of stimulating blood coagulation in a mammal, comprising administering to the mammal a PEGylated Factor IX conjugate according to  claim 6 . 
     
     
         11 . A method of treating hemophilia, comprising administering to a subject in need thereof an effective amount of a PEGylated Factor IX conjugate according to  claim 6 . 
     
     
         12 . The method according to  claim 9 , wherein hemophilia is hemophilia B.