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 - 264 . (canceled)
265 . A method for the prophylactic treatment of hemophilia A in a human subject in need thereof, comprising administering to the subject a therapeutic dose of a pharmaceutical composition, the pharmaceutical composition comprising:
(i) a combination of chimeric polypeptides comprising a first chimeric polypeptide and a second chimeric polypeptide, wherein the first chimeric polypeptide comprises a single chain Factor VIII (FVIII) and a second portion, and wherein the second chimeric polypeptide comprises two-chained FVIII and a second portion; and
(ii) at least one pharmaceutically acceptable excipient,
wherein 10% to 40% of the combination of chimeric polypeptides is the first chimeric polypeptide, and 60% to 90% combination of chimeric polypeptides is the second chimeric polypeptide,
wherein the single chain FVIII comprises a FVIII heavy chain and a FVIII light chain on a single polypeptide chain, and the two-chained FVIII comprises a FVIII heavy chain and a FVIII light chain on two polypeptide chains,
wherein the single chain FVIII comprises an amino acid sequence at least 95% identical to amino acids 20 to 1457 of SEQ ID NO: 2,
wherein the second portion of the first chimeric polypeptide comprises a first Fc region, wherein the first Fc region comprises an amino acid sequence that is at least 95% identical to amino acids 1458 to 1684 of SEQ ID NO: 2,
wherein the second portion of the second chimeric polypeptide comprises a first Fc region, wherein the first Fc region comprises an amino acid sequence that is at least 95% identical to amino acids 1458 to 1684 of SEQ ID NO: 2, and
wherein the chimeric polypeptide is administered at a dosing interval of between about 3 to 5 days.
266 . The method of claim 265 , wherein:
(i) 10% of the combination of chimeric polypeptides is the first chimeric polypeptide, and 90% of the combination of chimeric polypeptides is the second chimeric polypeptide;
(ii) 15% of the FVIII of the combination of chimeric polypeptides is the first chimeric polypeptide, and 85% of the combination of chimeric polypeptides is the second chimeric polypeptide;
(iii) 20% of the FVIII of the combination of chimeric polypeptides is the first chimeric polypeptide, and 80% of the combination of chimeric polypeptides is the second chimeric polypeptide;
(iv) 25% of the FVIII of the combination of chimeric polypeptides is the first chimeric polypeptide, and 75% of the combination of chimeric polypeptides is the second chimeric polypeptide;
(v) 30% of the FVIII of the combination of chimeric polypeptides is the first chimeric polypeptide, and 70% of the combination of chimeric polypeptides is the second chimeric polypeptide;
(vi) 35% of the FVIII of the combination of chimeric polypeptides is the first chimeric polypeptide, and 65% of the combination of chimeric polypeptides is the second chimeric polypeptide; or
(vii) 40% of the FVIII of the combination of chimeric polypeptides is the first chimeric polypeptide, and 60% of the combination of chimeric polypeptides is the second chimeric polypeptide.
267 . The method of claim 265 , wherein the two polypeptide chains of the two-chained FVIII are associated by a metal bond.
268 . The method of claim 265 , wherein the first Fc region of each of the chimeric polypeptides comprises an amino acid sequence that is at least 99% identical to amino acids 1458 to 1684 of SEQ ID NO: 2.
269 . The method of claim 268 , wherein the first Fc region of each of the chimeric polypeptides comprises an amino acid sequence identical to amino acids 1458 to 1683 of SEQ ID NO: 2.
270 . The method of claim 265 , wherein each of the chimeric polypeptides is a FVIIIFc monomer dimer hybrid, wherein the FVIIIFc monomer dimer hybrid comprises the FVIII covalently linked to the first Fc region, and wherein the first Fc region forms a disulfide bond with a second Fc region.
271 . The method of claim 270 , wherein the FVIII is covalently linked to the N-terminus of the first Fc region in each of the chimeric polypeptides.
272 . The method of claim 270 , wherein the second Fc region of each of the chimeric polypeptides comprises an amino acid sequence that is at least 95% identical to amino acids 21 to 247 of SEQ ID NO: 4.
273 . The method of claim 270 , wherein the second Fc region of each of the chimeric polypeptides comprises amino acids 21 to 246 of SEQ ID NO: 4.
274 . The method of claim 265 , wherein the FVIII of each of the chimeric polypeptides comprises amino acids 20 to 1457 of SEQ ID NO: 2.
275 . The method of claim 265 , wherein residue 1645 and/or residue 1648 corresponding to full-length mature FVIII is/are arginine in the single chain FVIII.
276 . The method of claim 265 , wherein residue 1645 and/or residue 1648 corresponding to full-length mature FVIII is/are substituted or mutated in the single chain FVIII compared to wild type FVIII.
277 . The method of claim 265 , wherein the hemophilia A is severe hemophilia A.
278 . A method for treating a subject in need of prophylactic treatment of hemophilia A, comprising administering to the subject a therapeutic dose of a pharmaceutical composition, the pharmaceutical composition comprising:
(i) a chimeric polypeptide, which comprises Factor VIII (FVIII), a first Fc region, and a second Fc region; and
(ii) at least one pharmaceutically acceptable excipient,
wherein:
15% to 40% of the FVIII of the chimeric polypeptide comprises single chain FVIII, and 60% to 85% of the FVIII of the chimeric polypeptide comprises two-chained FVIII, wherein the single chain FVIII comprises a FVIII heavy chain and a FVIII light chain on a single polypeptide chain, and the two-chained FVIII comprises a FVIII heavy chain and a FVIII light chain on two polypeptide chains;
wherein said pharmaceutical composition is lyophilized;
the FVIII comprises an amino acid sequence identical to amino acids 20 to 1457 of SEQ ID NO: 2;
the first Fc region comprises an amino acid sequence at least 95% identical to amino acids 1458 to 1684 of SEQ ID NO: 2;
the FVIII is covalently linked to the N-terminus of the first Fc region;
the first Fc region forms a disulfide bond with the second Fc region; and
the amino acid sequence of the second Fc region is identical to the amino acid sequence of the first Fc region.
279 . The method of claim 278 , wherein the first Fc region comprises an amino acid sequence at least 99% identical to amino acids 1458 to 1684 of SEQ ID NO: 2.
280 . The method of claim 279 , wherein the first Fc region comprises an amino acid sequence identical to amino acids 1458 to 1683 of SEQ ID NO: 2.
281 . The method of claim 278 , wherein the hemophilia A is severe hemophilia A.