Factor VIII compositions and methods
The present invention provides methods of increasing the half-life and/or specific activity of factor VIII. More specifically, the invention provides methods of increasing the half-life and/or specific activity of factor VIII by substituting one or more amino acids in the A2 domain. It further provides methods for producing such factor VIII mutants. The invention also provides polynucleotides encoding the mutant factor VIII, and methods of treating hemophilia using the polypeptides and polynucleotides of the invention.
1. A mutant factor VIII comprising an amino acid substitution at one or more positions selected from the group consisting of a position corresponding to Lys(377) and Lys(466) of SEQ ID NO:5, and optionally at one or more positions selected from the group consisting of a substitution at a position corresponding to Lys(380), Ser(488), Arg(489), Arg(490), Leu(491), Lys(493), Lys(496), His(497), Lys(499), Lys(512), Lys(523), and Lys(556) of SEQ ID NO:5, wherein the mutant factor VIII has procoagulant activity.
2. A pharmaceutically acceptable composition comprising the mutant factor VIII of claim 1 .
3. A method of treating hemophilia comprising administering to a patient in need thereof an effective amount of the composition of claim 2 .
4. The mutant factor VIII of claim 1 , further comprising an amino acid substitution at one or more additional positions in the A2 domain.
5. A pharmaceutically acceptable composition comprising the mutant factor VIII of claim 4 .
6. A method of treating hemophilia comprising administering to a patient in need thereof an effective amount of the composition of claim 5 .
7. A mutant factor VIII comprising an amino acid substitution at one or more positions selected from the group consisting of a position corresponding to Lys(377) and Lys(466) of SEQ ID NO:2, and optionally at one or more substitutions selected from the group consisting of a substitution at a position corresponding to Lys(380), Ser(488), Arg(489), Arg(490), Leu(491), Lys(493), Lys(496), His(497), Lys(499), Lys(512), Lys(523), and Lys(556) of SEQ ID NO:2, wherein the mutant factor VIII has procoagulant activity.
8. A pharmaceutically acceptable composition comprising the mutant factor VIII of claim 7 .
9. A method of treating hemophilia comprising administering to a patient in need thereof an effective amount of the composition of claim 8 .
10. The mutant factor VIII of claim 7 , further comprising an amino acid substitution at one or more additional positions in the A2 domain.
11. A pharmaceutically acceptable composition comprising the mutant factor VIII of claim 10 .
12. A method of treating hemophilia comprising administering to a patient in need thereof an effective amount of the composition of claim 11 .