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 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 25% of the FVIII of the chimeric polypeptide is single chain FVIII and 75% 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 pharmaceutical composition of claim 1 , wherein (a) 25% of the FVIII of the chimeric polypeptide is single chain FVIII and 75% of the FVIII of the chimeric polypeptide is processed FVIII, (b) 20% of the FVIII of the chimeric polypeptide is single chain FVIII and 80% of the FVIII of the chimeric polypeptide is processed FVIII, or (c) 15% of the FVIII of the chimeric polypeptide is single chain FVIII and 85% of the FVIII of the chimeric polypeptide is processed FVIII.
3. The pharmaceutical composition of claim 1 , wherein the single chain FVIII in the chimeric polypeptide has FVIII activity comparable to processed FVIII.
4. The pharmaceutical composition of claim 1 , wherein the chimeric polypeptide is a long-acting FVIII polypeptide.
5. The pharmaceutical composition 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.
6. The pharmaceutical composition of claim 5 , wherein the chimeric polypeptide is a FVIIIFc monomer dimer hybrid.
7. The pharmaceutical composition of claim 1 , wherein the FVIII comprises B-domain deleted FVIII.
8. The pharmaceutical composition of claim 7 , 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.
9. The pharmaceutical composition of claim 4 , wherein the chimeric polypeptide has a half-life longer than a polypeptide consisting of the FVIII.
10. The pharmaceutical composition of claim 1 , which is suitable for subcutaneous, intradermal, intravascular, intravenous, intramuscular, spinal, intracranial, intrathecal, intraocular, periocular, intraorbital, intrasynovial or intraperitoneal injection to a human.
11. The pharmaceutical composition of claim 1 , which is lyophilized.
12. The pharmaceutical composition of claim 2 , which is lyophilized.
13. The pharmaceutical composition of claim 3 , which is lyophilized.
14. The pharmaceutical composition of claim 4 , which is lyophilized.
15. The pharmaceutical composition of claim 5 , which is lyophilized.
16. The pharmaceutical composition of claim 6 , which is lyophilized.
17. The pharmaceutical composition of claim 7 , which is lyophilized.
18. The pharmaceutical composition of claim 8 , which is lyophilized.
19. The pharmaceutical composition of claim 10 , which is lyophilized.
20. The pharmaceutical composition of claim 11 , 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.
21. The pharmaceutical composition of claim 11 , 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.
22. The pharmaceutical composition of claim 11 , 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.