Fibronectin based scaffold domain proteins that bind to PCSK9

• US 8,420,098 B2
• Filed: 04/13/2011
• Issued: 04/16/2013
• Est. Priority Date: 04/13/2010
• Status: Active Grant

• Current Assignee: Bristol-Myers Squibb Company
• Original Assignee: Bristol-Myers Squibb Company
• Inventors: Camphausen, Ray, Davis, Jonathan H., Cload, Sharon T., Denhez, Fabienne M., Saeed-Kothe, Amna, Lipovsek, Dasa, Lo, Ching-Hsiung Frederick, Low, Chee Meng, Miao, Bowman, Mitchell, Tracy S., Parker, Rex A., Rakestraw, Ginger C., Russo, Katie A., Sitkoff, Doree F.
• Primary Examiner(s): HADDAD, MAHER M
• Application Number:
  US13/085,864
• Publication Number:
  US 20120094909A1
• Time in Patent Office: 734 Days
• Field of Search: None
• US Class Current: 424/185.1
• CPC Class Codes: A61K 38/39 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
• CPC Class Codes: A61K 47/549 Sugars, nucleosides, nucleotides or nucleic acids
• 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 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 14/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C07K 2318/20 Antigen-binding scaffold molecules wherein the scaffold is not an immunoglobulin variable region or antibody mimetics
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C07K 2319/31 fusions, other than Fc, for prolonged plasma life, e.g. albumin
• CPC Class Codes: C07K 2319/70 containing domain for protein-protein interaction
• CPC Class Codes: C12N 9/6424 Serine endopeptidases (3.4.21)

Abstract

The present invention relates to fibronectin based scaffold domain proteins that bind PCSK9. The invention also relates to the use of the innovative proteins in therapeutic applications to treat atherosclerosis, hypercholesterolemia and other cholesterol related diseases. The invention further relates to cells comprising such proteins, polynucleotides encoding such proteins or fragments thereof, and to vectors comprising the polynucleotides encoding the innovative protein.

Claims

1. An isolated polypeptide comprising a fibronectin type III tenth (¹⁰Fn3) domain wherein the ¹⁰Fn3 has a BC, DE and FG loop, and wherein the BC loop comprises SEQ ID NO:

14, the DE comprises SEQ ID NO;

25 and the FG loop comprises SEQ ID NO;

37, and wherein the polypeptide binds proprotein convertase subtilisin kexin type (PCSK9).

2. The isolated polypeptide of claim 1, wherein the polypeptide binds PCSK9 with a K_D of less than 500 nM.

3. The isolated polypeptide of claim 1, further comprising one or more pharmacokinetic (PK) moieties selected from the group consisting of polyethylene glycol, sialic acid, Fc, Fc fragment, transferring, serum albumin, a serum albumin binding protein, and a serum immunoglobulin binding protein.

4. A pharmaceutically acceptable composition comprising the polypeptide of claim 1.

5. An isolated polypeptide comprising a ¹⁰Fn3 domain wherein the isolated polypeptide comprises SEQ ID NO:

82, and wherein the isolated polypeptide binds PCSK9.

6. The isolated polypeptide of claim 5, wherein the polypeptide binds PCSK9 with a K_D of less than 500 nM.

7. The isolated polypeptide of claim 5, further comprising one or more pharmacokinetic (PK) moieties selected from the group consisting of polyethylene glycol, sialic acid, Fc, Fc fragment, transferring, serum albumin, a serum albumin binding protein, and a serum immunoglobulin binding protein.

8. A pharmaceutically acceptable composition comprising the polypeptide of claim 5.