Methods of preparing pharmaceutical compositions for treatment of factor Vlll associated disorders and methods of use
The invention concerns glycosylated proteins having human factor VIII activity. In a preferred embodiment, the protein is glycosylated with oligosaccharides that include an alpha-(2,6)-linked sialic acid and a bisecting GlcNAc linked to a core beta-mannose.
1 - 24 . (canceled)
25 . An isolated glycosylated protein having factor VIII procoagulant activity, wherein the amino acid sequence of the protein has at least 62% identity with SEQ ID NO:1 and the protein comprises an N-linked oligosaccharide having an alpha-(2,6)-linked sialic acid.
26 . The isolated protein of claim 25 , wherein the amino acid sequence has at least 72% identity with SEQ ID NO:1.
27 . The isolated protein of claim 25 , wherein the amino acid sequence has at least 82% identity with SEQ ID NO:1.
28 . A pharmaceutical composition comprising the isolated protein of claim 25 and a pharmaceutically acceptable adjuvant.
29 . The pharmaceutical composition of claim 28 wherein the composition is substantially free of a toxic solubilizing agent.
30 . A method of treating a factor VIII-associated disorder comprising administering to a human in need thereof a therapeutically effective amount of the pharmaceutical composition of claim 28 .
31 . A method of reducing hemorrhage in a patient due to surgical procedure comprising administering to the patient a therapeutically effective amount of the pharmaceutical composition of claim 28 .
32 . The method of claim 31 , wherein the pharmaceutical composition is administered to the patient preoperatively.
33 . The method of claim 32 further comprising the step of re-administering the pharmaceutical composition 6 to 12 hours after the initial administration.
34 . An isolated glycosylated protein having factor VIII procoagulant activity, wherein the amino acid sequence of the protein has at least 62% identity with SEQ ID NO:1 and the protein comprises an N-linked oligosaccharide having a bisecting GlcNAc linked to a core beta-mannose.
35 . The isolated protein of claim 34 , wherein the amino acid sequence has at least 72% identity with SEQ ID NO:1.
36 . The isolated protein of claim 34 , wherein the amino acid sequence has at least 82% identity with SEQ ID NO:1.
37 . A pharmaceutical composition comprising the isolated protein of claim 34 and a pharmaceutically acceptable adjuvant.
38 . The pharmaceutical composition of claim 37 , wherein the composition is substantially free of a toxic solubilizing agent.
39 . A method of treating a factor VIII-associated disorder comprising administering to a human in need thereof a therapeutically effective amount of the pharmaceutical composition of claim 37 .
40 . A method of reducing hemorrhage in a patient due to surgical procedure comprising administering to the patient a therapeutically effective amount of the pharmaceutical composition of claim 37 .
41 . The method of claim 40 , wherein the pharmaceutical composition is administered to the patient preoperatively.
42 . The method of claim 41 further comprising the step of re-administering the pharmaceutical composition 6 to 12 hours after the initial administration.