Factor VIII-FC chimeric and hybrid polypeptides, and methods of use thereof
The present invention provides methods of administering Factor VIII; methods of administering chimeric and hybrid polypeptides comprising Factor VIII; chimeric and hybrid polypeptides comprising Factor VIII; polynucleotides encoding such chimeric and hybrid polypeptides; cells comprising such polynucleotides; and methods of producing such chimeric and hybrid polypeptides using such cells.
1. A method of reducing the incidence of a bleeding episode in a human subject with hemophilia A, comprising
administering to the human subject multiple doses of a chimeric polypeptide comprising a Factor VIII portion and an Fc portion at a dosing interval of about three to about five days, wherein each of the multiple doses is about 25-100 IU/kg, wherein the chimeric polypeptide is administered intravenously.
2. The method of claim 1 , wherein said dosing interval of said chimeric polypeptide is about every five days.
3. The method of claim 1 , wherein said dosing interval of said chimeric polypeptide is about once every 4 days.
4. The method of claim 1 , wherein the Factor VIII portion of said chimeric polypeptide is at least 90% or 95% identical to a Factor VIII amino acid sequence with a signal sequence selected from the group consisting of amino acids 1 to 1457 of SEQ ID NO:2; amino acids 1 to 2351 of SEQ ID NO:6; amino acids 1 to 759 of SEQ ID NO:8; amino acids 1 to 764 of SEQ ID NO:10; and amino acids 1 to 704 of SEQ ID NO:12 or at least 90% or 95% identical to a Factor VIII amino acid sequence without a signal sequence selected from the group consisting of amino acids 20 to 1457 of SEQ ID NO:2; amino acids 20 to 2351 of SEQ ID NO:6; amino acids 20 to 759 of SEQ ID NO:8; amino acids 20 to 764 of SEQ ID NO:10; and amino acids 21 to 704 of SEQ ID NO:12, and
wherein said chimeric polypeptide is in the form of a hybrid comprising a second polypeptide in association with said chimeric polypeptide, wherein said second polypeptide consists essentially of an Fc.
5. The method of claim 4 , wherein said chimeric polypeptide comprises a sequence at least 90% or 95% identical to the Factor VIII and Fc amino acid sequence without a signal sequence set forth as amino acids 20 to 1684 of SEQ ID NO:2 or at least 90% or 95% identical to the Factor VIII and Fc amino acid sequence with a signal sequence set forth as amino acids 1 to 1684 of SEQ ID NO:2.
6. A method for prophylactic treatment of a spontaneous bleeding episode in a human subject with hemophilia A, comprising
administering to the subject multiple doses of a chimeric polypeptide comprising a Factor VIII portion and an Fc portion at a dosing interval of about three to about five days, wherein each of the multiple doses is about 25-65 IU/kg, wherein the chimeric polypeptide is administered intravenously.
7. The method of claim 6 , wherein said Factor VIII portion is at least 90% or 95% identical to a Factor VIII amino acid sequence without a signal sequence selected from the group consisting of amino acids 20 to 1457 of SEQ ID NO:2; amino acids 20 to 2351 of SEQ ID NO:6; amino acids 20 to 759 of SEQ ID NO:8; amino acids 20 to 764 of SEQ ID NO:10; and amino acids 21 to 704 of SEQ ID NO:12.
8. The method of claim 6 , wherein said Factor VIII portion is at least 90% or 95% identical to a Factor VIII amino acid sequence with a signal sequence selected from the group consisting of amino acids 1 to 1457 of SEQ ID NO:2; amino acids 1 to 2351 of SEQ ID NO:6; amino acids 1 to 759 of SEQ ID NO:8; amino acids 1 to 764 of SEQ ID NO:10; and amino acids 1 to 704 of SEQ ID NO:12, and
wherein said Fc portion is at least 90%, at least 95%, or 100% identical to an Fc amino acid sequence selected from the group consisting of amino acids 1458 to 1684 of SEQ ID NO:2;
amino acids 2352 to 2578 of SEQ ID NO:6; amino acids 760 to 986 of SEQ ID NO:8; amino acids 765 to 991 of SEQ ID NO:10; and amino acids 705 to 931 of SEQ ID NO:12.
9. The method of claim 6 , wherein said chimeric polypeptide is a Factor VIII-Fc chimeric polypeptide in the form of a hybrid comprising a second polypeptide in association with said chimeric polypeptide, wherein said second polypeptide consists essentially of an Fc, wherein said chimeric polypeptide comprises a sequence at least 95% identical to the Factor VIII and Fc amino acid sequence without a signal sequence set forth as amino acids 20 to 1684 of SEQ ID NO:2 or at least 90% or 95% identical to the Factor VIII and Fc amino acid sequence with a signal sequence set forth as amino acids 1 to 1684 of SEQ ID NO:2.
10. The method of claim 1 , wherein the chimeric polypeptide comprises a FVIII portion and two Fcs, wherein one of the Fcs is fused to the C-terminus of the light chain of the FVIII portion.
11. The method of claim 1 , wherein the chimeric polypeptide is a rFVIIIFc monomer dimer hybrid recombinant fusion protein comprising a single molecule of recombinant B-domain deleted human FVIII fused to the dimeric Fc domain of human IgG1, with no intervening linker sequence.
12. The method of claim 1 , wherein the hemophilia A is severe hemophilia A.
13. The method of claim 1 , wherein each of the multiple doses is about 70-80 IU/kg.
14. The method of claim 1 , wherein each of the multiple doses is about 80-90 IU/kg.
15. The method of claim 1 , wherein each of the multiple doses is about 90-100 IU/kg.
16. The method of claim 6 , wherein the chimeric polypeptide comprises a FVIII portion and two Fcs, wherein one of the Fcs is fused to the C-terminus of the light chain of the FVIII portion.
17. The method of claim 6 , wherein the chimeric polypeptide is a rFVIIIFc monomer dimer hybrid recombinant fusion protein comprising a single molecule of recombinant B-domain deleted human FVIII fused to the dimeric Fc domain of human IgG1, with no intervening linker sequence.
18. The method of claim 6 , wherein the hemophilia A is severe hemophilia A.
19. The method of claim 6 , wherein each of the multiple doses is about 70-80 IU/kg.
20. The method of claim 6 , wherein each of the multiple doses is about 80-90 IU/kg.
21. The method of claim 6 , wherein each of the multiple doses is about 90-100 IU/kg.
22. A method for treating a bleeding episode in a human subject with hemophilia A, comprising
administering to the subject multiple doses of a chimeric polypeptide comprising a Factor VIII portion and an Fc portion at a dosing interval of about three to about five days, wherein each of the multiple doses is about 25-80 IU/kg, wherein the chimeric polypeptide is administered intravenously.
23. The method of claim 22 , wherein said Factor VIII portion is at least 90% or 95% identical to a Factor VIII amino acid sequence without a signal sequence selected from the group consisting of amino acids 20 to 1457 of SEQ ID NO:2; amino acids 20 to 2351 of SEQ ID NO:6; amino acids 20 to 759 of SEQ ID NO:8; amino acids 20 to 764 of SEQ ID NO:10; and amino acids 21 to 704 of SEQ ID NO:12.
24. The method of claim 22 , wherein said Factor VIII portion is at least 90% or 95% identical to a Factor VIII amino acid sequence with a signal sequence selected from the group consisting of amino acids 1 to 1457 of SEQ ID NO:2; amino acids 1 to 2351 of SEQ ID NO:6; amino acids 1 to 759 of SEQ ID NO:8; amino acids 1 to 764 of SEQ ID NO:10; and amino acids 1 to 704 of SEQ ID NO:12, and
wherein said Fc portion is at least 90%, at least 95%, or 100% identical to an Fc amino acid sequence selected from the group consisting of amino acids 1458 to 1684 of SEQ ID NO:2; amino acids 2352 to 2578 of SEQ ID NO:6; amino acids 760 to 986 of SEQ ID NO:8; amino acids 765 to 991 of SEQ ID NO:10; and amino acids 705 to 931 of SEQ ID NO:12.
25. The method of claim 22 , wherein said chimeric polypeptide is a Factor VIII-Fc chimeric polypeptide in the form of a hybrid comprising a second polypeptide in association with said chimeric polypeptide, wherein said second polypeptide consists essentially of an Fc, wherein said chimeric polypeptide comprises a sequence at least 95% identical to the Factor VIII and Fc amino acid sequence without a signal sequence set forth as amino acids 20 to 1684 of SEQ ID NO:2 or at least 90% or 95% identical to the Factor VIII and Fc amino acid sequence with a signal sequence set forth as amino acids 1 to 1684 of SEQ ID NO:2.
26. The method of claim 22 , wherein the chimeric polypeptide comprises a FVIII portion and two Fcs, wherein one of the Fcs is fused to the C-terminus of the light chain of the FVIII portion.
27. The method of claim 22 , wherein the chimeric polypeptide is a rFVIIIFc monomer dimer hybrid recombinant fusion protein comprising a single molecule of recombinant B-domain deleted human FVIII fused to the dimeric Fc domain of human IgG1, with no intervening linker sequence.
28. The method of claim 22 , wherein the hemophilia A is severe hemophilia A.
29. The method of claim 1 , wherein the Factor VIII portion is pegylated Factor VIII.
30. The method of claim 1 , wherein each of the multiple doses is 25-65 IU/kg.
31. The method of claim 1 , wherein each of the multiple doses is 30-50 IU/kg.
32. The method of claim 1 , wherein each of the multiple doses is 30-65 IU/kg.
33. The method of claim 1 , wherein each of the multiple doses is 50 IU/kg.
34. The method of claim 1 , wherein the dosing interval is every three days.
35. The method of claim 6 , wherein the Factor VIII portion is pegylated Factor VIII.
36. The method of claim 6 , wherein each of the multiple doses is 25-65 IU/kg.
37. The method of claim 6 , wherein each of the multiple doses is 30-50 IU/kg.
38. The method of claim 6 , wherein each of the multiple doses is 30-65 IU/kg.
39. The method of claim 6 , wherein each of the multiple doses is 50 IU/kg.
40. The method of claim 6 , wherein the dosing interval is every three days.
41. The method of claim 22 , wherein the Factor VIII portion is pegylated Factor VIII.
42. The method of claim 22 , wherein each of the multiple doses is 25-65 IU/kg.
43. The method of claim 22 , wherein each of the multiple doses is 30-50 IU/kg.
44. The method of claim 22 , wherein each of the multiple doses is 30-65 IU/kg.
45. The method of claim 22 , wherein each of the multiple doses is 50 IU/kg.
46. The method of claim 22 , wherein the dosing interval is every three days.
47. The method of claim 6 , wherein each of the multiple doses is 50 IU/kg, and wherein the dosing interval is about every 4 days.
48. The method of claim 47 , wherein the human subject with hemophilia A is at least 12 years old.
49. The method of claim 22 , wherein each of the multiple doses is 80 IU/kg.
50. The method of claim 1 , wherein the chimeric protein further comprises an intervening linker sequence between the Factor VIII portion and the Fc portion.
51. The method of claim 50 , wherein the Fc portion comprises a Pro331Ser substitution.
52. The method of claim 50 , wherein the Fc portion is a human immunoglobulin G1 (IgG1) Fc portion comprising three amino acid substitutions that do not cause a significant loss of Fc binding affinity to FcRn.
53. The method of claim 50 , wherein the long-acting Factor VIII is pegylated.
54. The method of claim 50 , wherein the Fc portion is a human immunoglobulin G2 (IgG2) Fc portion comprising three amino acid substitutions that do not cause a significant loss of Fc binding affinity to FcRn.
55. The method of claim 6 , wherein the chimeric protein further comprises an intervening linker sequence between Factor VIII portion and the Fc portion.
56. The method of claim 55 , wherein the Fc portion comprises a Pro331Ser substitution.
57. The method of claim 55 , wherein the Fc portion is a human immunoglobulin G1 (IgG1) Fc portion comprising three amino acid substitutions that do not cause a significant loss of Fc binding affinity to FcRn.
58. The method of claim 55 , wherein the long-acting Factor VIII is pegylated.
59. The method of claim 55 , wherein the Fc portion is a human immunoglobulin G2 (IgG2) Fc portion comprising three amino acid substitutions that do not cause a significant loss of Fc binding affinity to FcRn.
60. The method of claim 22 , wherein the chimeric protein further comprises an intervening linker sequence between the Factor VIII portion and the Fc portion.
61. The method of claim 60 , wherein the Fc portion comprises a Pro331Ser substitution.
62. The method of claim 60 , wherein the Fc portion is a human immunoglobulin G1 (IgG1) Fc portion comprising three amino acid substitutions that do not cause a significant loss of Fc binding affinity to FcRn.
63. The method of claim 60 , wherein the long-acting Factor VIII is pegylated.
64. The method of claim 60 , wherein the Fc portion is a human immunoglobulin G2 (IgG2) Fc portion comprising three amino acid substitutions that do not cause a significant loss of Fc binding affinity to FcRn.