Factor VIII chimeric and hybrid polypeptides, and methods of use thereof
The present invention provides methods of administering Factor VIII (processed FVIII, single chain FVIII, or a combination thereof); 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 treating hemophilia A in a subject in need thereof, comprising administering to the subject an effective amount of a pharmaceutical composition comprising (i) a chimeric polypeptide, which comprises Factor VIII (FVIII) and a second portion, and (ii) at least one pharmaceutically acceptable excipient, wherein 15% to 40% of the FVIII of the chimeric polypeptide is single chain FVIII and 60% to 85% of the FVIII of the chimeric polypeptide is processed FVIII, wherein the single chain FVIII comprises a FVIII heavy chain and a FVIII light chain on a single polypeptide chain and the processed FVIII comprises a FVIII heavy chain and a FVIII light chain on two polypeptide chains.
2. The method of claim 1 , wherein 15% to 25% of the FVIII of the chimeric polypeptide is single chain Factor VIII.
3. The method of claim 1 , wherein 20% to 30% of the FVIII of the chimeric polypeptide is single chain Factor VIII.
4. The method of claim 1 , wherein 25% to 35% of the FVIII of the chimeric polypeptide is single chain Factor VIII.
5. The method of claim 1 , wherein 30% to 40% of the FVIII of the chimeric polypeptide is single chain Factor VIII.
6. The method of claim 1 , wherein 15% of the FVIII of the chimeric polypeptide is single chain FVIII and 85% of the FVIII of the chimeric polypeptide is processed FVIII.
7. The method of claim 1 , wherein 20% of the FVIII of the chimeric polypeptide is single chain FVIII and 80% of the FVIII of the chimeric polypeptide is processed FVIII.
8. The method of claim 1 , wherein 25% of the FVIII of the chimeric polypeptide is single chain FVIII and 75% of the FVIII of the chimeric polypeptide is processed FVIII.
9. The method of claim 1 , wherein 30% of the FVIII of the chimeric polypeptide is single chain FVIII and 70% of the FVIII of the chimeric polypeptide is processed FVIII.
10. The method of claim 1 , wherein 35% of the FVIII of the chimeric polypeptide is single chain FVIII and 65% of the FVIII of the chimeric polypeptide is processed FVIII.
11. The method of claim 1 , wherein 10% of the FVIII of the chimeric polypeptide is single chain FVIII and 60% of the FVIII of the chimeric polypeptide is processed FVIII.
12. The method of claim 1 , wherein the second portion is selected from the group consisting of an Fc region, albumin, a PAS sequence, transferrin, CTP (28 amino acid C-terminal peptide (CTP) of hCG with its 4 O-glycans), polyethylene glycol (PEG), hydroxyethyl starch (HES), albumin binding polypeptide, albumin-binding small molecules, and two or more combinations thereof.
13. The method of claim 1 , wherein the chimeric polypeptide is a FVIIIFc monomer dimer hybrid.
14. The method of claim 1 , wherein the FVIII comprises B-domain deleted FVIII.
15. The method of claim 14 , wherein residue 1645 corresponding to full-length, mature FVIII, residue 1648 corresponding to full-length, mature FVIII, or both residues in the single chain FVIII is arginine.
16. The method of claim 1 , wherein the pharmaceutical composition is administered by intravenous injection.
17. The method of claim 1 , wherein the pharmaceutical composition is lyophilized prior to formulation in an aqueous solution.
18. The method of claim 1 , which is a method of prophylactic treatment.
19. The method of claim 1 , which is a method of on-demand treatment.
20. The method of claim 1 , wherein the FVIII portion comprises an amino acid sequence at least 95% identical to amino acids 20 to 1457 of SEQ ID NO: 2.
21. The method of claim 20 , wherein the FVIII portion comprises an amino acid sequence identical to amino acids 20 to 1457 of SEQ ID NO: 2.
22. The method of claim 1 , wherein the second portion comprises an Fc region.
23. The method of claim 1 , wherein the second portion comprises an amino acid sequence at least 95% identical to amino acids 21 to 247 of SEQ ID NO: 4.
24. The method of claim 23 , wherein the second portion comprises an amino acid sequence identical to amino acids 21 to 246 of SEQ ID NO: 4.
25. A method of decreasing the incidence of spontaneous bleeding in a subject who has hemophilia A, comprising administering to the subject an effective amount of a pharmaceutical composition comprising (i) a FVIIIFc monomer dimer hybrid comprising a single molecule of recombinant B-domain deleted human FVIII (BDD-rFVIII) fused to the N-terminus of a polypeptide of a dimeric human IgG1 Fc domain with no intervening linker sequence, and (ii) at least one pharmaceutically acceptable excipient,
wherein 15% to 40% of the BDD-rFVIII is single chain BDD-rFVIII, and wherein 60% to 85% of the BDD-rFVIII is processed BDD-rFVIII;
wherein said BDD-rFVIII comprises an amino acid sequence at least 95% identical to amino acids 20 to 1457 of SEQ ID NO: 2, and
wherein each polypeptide of the dimeric human IgG1 Fc domain comprises an amino acid sequence at least 95% identical to amino acids 21 to 247 of SEQ ID NO: 4.
26. The method of claim 25 , wherein the hemophilia A is severe hemophilia A.