Glycopegylated Factor IX
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.