Factor VIII conjugates
This invention relates to Factor VIII muteins that are covalently bound, at a predefined site that is not an N-terminal amine, to one or more biocompatible polymers such as polyethylene glycol. The mutein conjugates retain FVIII procoagulant activity and have improved pharmacokinetic properties.
1. An isolated polypeptide conjugate comprising a functional factor VIII polypeptide and one or more biocompatible polymers, wherein the functional factor VIII polypeptide comprises the amino acid sequence of SEQ ID NO: 4 or an allelic variant thereof and has a B-domain, and further wherein the biocompatible polymer comprises polyalkylene oxide and is covalently attached to the functional factor VIII polypeptide at the B-domain.
2. The isolated polypeptide conjugate of claim 1 , wherein the biocompatible polymer comprises polyethylene glycol.
3. The isolated polypeptide conjugate of claim 2 , wherein the polyethylene glycol comprises methoxypolyethylene glycol.
4. The isolated polypeptide conjugate of claim 3 , wherein the methoxypolyethylene glycol has a size range of from 3 kD to 100 kD.
5. The isolated polypeptide conjugate of claim 3 , wherein the methoxypolyethylene glycol has a size range of from 5 kD to 64 kD.
6. The isolated polypeptide conjugate of claim 3 , wherein the methoxypolyethylene glycol has a size range of from 5 kD to 43 kD.
7. The isolated polypeptide conjugate of claim 2 , wherein the polyethylene glycol has a size of 5, 12, 22, 30, or 43 kD.
8. A pharmaceutical composition comprising a therapeutically effective amount of the isolated polypeptide conjugate of claim 1 and a pharmaceutically acceptable adjuvant.
9. A pharmaceutical composition comprising (1) a therapeutically effective amount of a monopegylated polypeptide conjugate, wherein the monopegylated polypeptide conjugate comprises a functional factor VIII polypeptide and one polyethylene glycol polymer, the functional factor VIII polypeptide comprises the amino acid sequence of SEQ ID NO: 4 or an allelic variant thereof and has a B-domain, and the polyethylene glycol is covalently attached to the functional factor VIII polypeptide at the B-domain; and (2) a pharmaceutically acceptable adjuvant.