Conjugated factor VIII molecules

• US 9,150,848 B2
• Filed: 05/08/2014
• Issued: 10/06/2015
• Est. Priority Date: 02/27/2008
• Status: Active Grant

• Current Assignee: Novo Nordisk A/S
• Original Assignee: Novo Nordisk A/S
• Inventors: Bolt, Gert, Vandahl, Brian Berg Stidsen, Thim, Lars, Stennicke, Henning Ralf, Steenstrup, Thomas Dock, DeFrees, Shawn
• Primary Examiner(s): Skowronek, Karlheinz R
• Assistant Examiner(s): Fischer, Joseph
• Application Number:
  US14/272,726
• Publication Number:
  US 20140242057A1
• Time in Patent Office: 516 Days
• Field of Search: None
• US Class Current: 1/1
• 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: A61K 38/37 Factors VIII
• CPC Class Codes: A61K 47/26 Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• 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 relates to B-domain truncated Factor VIII molecules with a modified circulatory half life, said molecule being covalently conjugated with a hydrophilic polymer. The invention furthermore relates to methods for obtaining such molecules as well as use of such molecules.

Claims

1. A B-domain truncated Factor VIII (FVIII) molecule, comprising:

a heavy chain, a light chain, and a hydrophilic polymer;

wherein (1) the heavy chain comprises a linker derived from the B-domain of a full-length FVIII molecule (SEQ ID NO;

1) comprising a protease recognition site, (2) the hydrophilic polymer is covalently conjugated to the linker at Ser750 amino acid residue numbered relative to SEQ ID NO;

1 via an O-linked oligosaccharide, (3) the length of the linker is 20-30 amino acids, and (4) activation of the molecule by thrombin results in removal of the linker and the hydrophilic polymer;

wherein the B-domain truncated FVIII molecule has an increased circulatory half-life compared to a wild-type FVIII molecule.

2. The molecule according to claim 1, wherein the hydrophilic polymer is polyethylene glycol (PEG).

3. The molecule according to claim 2, wherein the size of the PEG is in a range of 10,000 to 160,000 Da.

4. The molecule according to claim 3, wherein the size of the PEG is 40,000 Da.

5. The molecule according to claim 1, wherein the hydrophilic polymer is a polysaccharide.

6. The molecule according to claim 5, wherein the polysaccharide is polysialic acid.

7. A pharmaceutical composition comprising a molecule according to claim 1.

8. A method of making a molecule according to claim 1, wherein said method comprises conjugating a B-domain truncated Factor VIII molecule with a hydrophilic polymer via an O-linked oligosaccharide in the truncated B domain.

9. A molecule obtained by a method according to claim 8.

10. A method of treating hemophilia comprising administering to a patient in need of such treatment a molecule according to claim 1.

11. A method according to claim 10, wherein said molecule is administered subcutaneously.

12. A method according to claim 10, wherein said molecule is administered intravenously.