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 . A PEGylated Factor IX protein comprising SEQ ID NO:1 and a poly(ethylene glycol) moiety, wherein the poly(ethylene glycol) moiety is conjugated to an amino acid selected from Asn 157 and Asn 167 of SEQ ID NO:1 via a glycosyl linking group of a glycosyl residue, and having a composition according to 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. 
 
 
 
       
     
     
         2 . A pharmaceutical formulation comprising the Factor IX conjugate of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         3 . A method of stimulating blood coagulation in a mammal, said method comprising administering to said mammal the PEGylated Factor IX conjugate of  claim 1 . 
     
     
         4 . A method of treating hemophilia in a subject, said method comprising administering to said subject said Factor IX conjugate of  claim 1 . 
     
     
         5 . The method of treating hemophilia of  claim 4 , wherein hemophilia is hemophilia B.