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 pharmaceutical composition comprising (i) a conjugate of a Factor IX variant of SEQ ID NO:1 and a modified sugar and (ii) a pharmaceutically acceptable excipient;
 wherein the modified sugar comprises the formula   
       
         
           
           
               
               
           
         
       
       and
 wherein
 R 10  is a link to a —O-glycosyl residue or an amino acid of the Factor IX variant; 
 R 11  is selected from the group consisting of OH and NHC(O)CH 3 ; 
 R 12  is selected from the group consisting of H, C(O)CH 3 , NH, and NHC(O)CH 3 ; 
 R 13  is selected from the group consisting of OH and C(O)CH 3 ; and 
 R 14  is selected from the group consisting of H, C(O)CH 3 , NH, and NHC(O)CH 3 . 
 
 
     
     
         2 . The pharmaceutical composition according to  claim 1 , wherein the Factor IX variant comprises an alanine in the first position. 
     
     
         3 . A method of stimulating blood coagulation in a mammal, comprising administering to said mammal the pharmaceutical composition according to  claim 1 . 
     
     
         4 . A method of treating hemophilia in a subject, comprising administering to said subject the pharmaceutical composition according to  claim 1 . 
     
     
         5 . The method according to  claim 4 , wherein the hemophilia is hemophilia B. 
     
     
         6 . A method of preparing the conjugate of the pharmaceutical composition according to  claim 1 , comprising conjugating the Factor IX variant and the modified sugar via chemoPEGylation.