FACTOR VIII COMPOSITIONS AND METHODS

• US 20100167392A1
• Filed: 10/02/2009
• Published: 07/01/2010
• Est. Priority Date: 10/31/2003
• Status: Abandoned Application

• Current Assignee: University of Maryland Baltimore County (University of Maryland)
• Original Assignee: University of Maryland Baltimore County (University of Maryland)
• Inventors: Ananyeva, Natalya M., SAENKO, Evgueni L., Sarafanov, Andrey G.
• Application Number:
  US12/572,363
• Publication Number:
  US 20100167392A1
• Time in Patent Office: Days
• US Class Current: 435/325
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C07K 14/755 Factors VIII , e.g. factor VIII C (AHF), factor VIII Ag (VWF)

Abstract

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.

Claims

1-4. -4. (canceled)

5. An isolated polynucleotide encoding a mutant factor VIII comprising the amino acid sequence of SEQ ID NO:

5, further comprising an amino acid substitution at position Lys(466) and/or position Arg(471) and optionally at one or more positions selected from the group consisting of Lys(380), Arg(484), Tyr(487), Ser(488), Arg(489), Arg(490), His(497), His(499), Lys(523), and Lys(556).

6-10. -10. (canceled)

11. An isolated polynucleotide encoding a mutant factor VIII comprising the amino acid sequence of SEQ ID NO:

5, further comprising an amino acid substitution at position Lys(466) and/or position Arg(471) and optionally at one or more positions selected from the group consisting of Lys(380), Arg(484), Tyr(487), Ser(488), Arg(489), Arg(490), His(497), His(499), Lys(523), and Lys(556), further comprising an amino acid substitution at one or more additional positions in the A2 domain.

33. The polynucleotide of any one of claims 5, 11, 17 or 23 wherein said polynucleotide is in a vector.

34. The polynucleotide of claim 33, wherein said vector is an expression vector capable of expressing said mutant factor VIII.

35. The polynucleotide of any one of claims 5, 11, 17 or 23 wherein said polynucleotide is a genetic construct operatively linked to a promoter sequence and to a polyadenylation signal sequence.

36. The polynucleotide of any one of claims 5, 11, 17 or 23 wherein said polynucleotide is in a host cell.

12-16. -16. (canceled)

17. A polynucleotide encoding a mutant factor VIII comprising the amino acid sequence of SEQ ID NO:

5, further comprising a substitution at one or more positions selected from the group consisting of Lys(377), His(378), and Lys(466), and optionally at one or more positions selected from the group consisting of Lys (380), Ser(488), Arg(489), Arg(490), Leu(491), Lys(493), Lys(496), His(497), His(499), Lys(512), Lys(523), and Lys(556).

18-22. -22. (canceled)

23. A polynucleotide encoding a mutant factor VIII comprising the amino acid sequence of SEQ ID NO:

5, further comprising a substitution at one or more positions selected from the group consisting of Lys(377), His(378), and Lys(466), and optionally at one or more positions selected from the group consisting of Lys(380), Ser(488), Arg(489), Arg(490), Leu(491), Lys(493), Lys(496), His(497), His(499), Lys(512), Lys(523), and Lys(556), further comprising the amino acid substitution at one or more additional positions in the A2 domain.

24-32. -32. (canceled)