Methods and compositions for reducing heparan sulfate proteoglycan-mediated clearance of factor VIII

• US 7,615,622 B2
• Filed: 01/11/2002
• Issued: 11/10/2009
• Est. Priority Date: 01/12/2001
• Status: Expired due to Fees

• Current Assignee: University of Maryland Baltimore County (University of Maryland)
• Original Assignee: University of Maryland Baltimore County (University of Maryland)
• Inventors: Saenko, Evgueni L., Sarafanov, Andrey G
• Primary Examiner(s): CHEN, SHIN LIN
• Application Number:
  US10/472,516
• Publication Number:
  US 20040248785A1
• Time in Patent Office: 2,860 Days
• Field of Search: 536/23.1, 536/23.5, 530/350
• US Class Current: 536/23.5
• CPC Class Codes: A61K 2039/53 DNA (RNA) vaccination
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/04 Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
• CPC Class Codes: C07K 14/755 Factors VIII , e.g. factor VIII C (AHF), factor VIII Ag (VWF)
• CPC Class Codes: C12N 9/96 Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates

Abstract

The present invention provides methods of increasing the half-life factor VIII. More specifically, the invention methods of increasing the half-life of factor VIII by substituting amino acids in the A2 domain or in the C2 domain of factor VIII or in both domains. It further provides factor VIII mutants produced by these methods. The invention also provides a method of using receptor-associated protein (RAP) to increase the half-life of factor VIII. The invention also provides polynucleotides encoding the mutant factor VIII, polynucleotides encoding RAP, and methods of treating hemophilia using the polypeptides and polynucleotides of the invention.

Claims

1. A isolated polynucleotide encoding a mutant factor VIII polypeptide, wherein said mutant factor VIII polypeptide consists of a human or porcine factor VIII polypeptide containing at least one nonconservative amino acid substitution at a position corresponding to an amino acid selected from the group consisting of Lys (380), Lys (512), Lys (523), Lys (556), Lys (570), Arg (571), and Lys (659) of SEQ ID NO:

5, and wherein said mutant factor VIII polypeptide exhibits procoagulant activity and reduced heparan sulfate proteoglycan (HSPG)-dependent, receptor-independent clearance.

2. A isolated polynucleotide encoding a mutant factor VIII polypeptide, wherein said mutant factor VIII polypeptide consists of a human or porcine factor VIII polypeptide containing a nonconservative amino acid substitution at a position corresponding to the amino acid at position Arg (490) of SEQ ID NO:

5, and at least one nonconservative amino acid substitution at a position corresponding to an amino acid selected from the group consisting of Lys (380), Lys (512), Lys (523), Lys (556), Arg (562), Lys (570), Arg (571), and Lys (659) of SEQ ID NO;

5, and wherein said mutant factor VIII polypeptide exhibits procoagulant activity and reduced heparan sulfate proteoglycan (HSPG)-dependent, receptor-independent clearance.

3. A isolated polynucleotide encoding a mutant factor VIII polypeptide, wherein said mutant factor VIII polypeptide consists of a human or porcine factor VIII polypeptide containing two or more nonconservative amino acid substitutions at a position corresponding to an amino acid selected from the group consisting of Lys (380), Lys (512), Lys (523), Lys (556), Arg(562), Lys (570), Arg (571), and Lys (659) of SEQ ID NO:

5, and wherein said mutant factor VIII polypeptide exhibits procoagulant activity and reduced heparan sulfate proteoglycan (HSPG)-dependent, receptor-independent clearance.