Albumin fusion proteins

• US 7,569,384 B2
• Filed: 08/08/2006
• Issued: 08/04/2009
• Est. Priority Date: 02/09/2004
• Status: Expired due to Fees

• Current Assignee: Human Genome Sciences Incorporated (GSK plc)
• Original Assignee: Human Genome Sciences Incorporated (GSK plc)
• Inventors: Rosen, Craig A., Haseltine, William A., Ruben, Steven M.
• Primary Examiner(s): Nashed; Nashaat T
• Assistant Examiner(s): Moore; William W
• Application Number:
  US11/500,508
• Publication Number:
  US 20070027306A1
• Time in Patent Office: 1,092 Days
• Field of Search: None
• US Class Current: 435/320.1
• 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: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 37/08 Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/00 Drugs for disorders of the endocrine system
• CPC Class Codes: C07K 14/765 Serum albumin, e.g. HSA
• CPC Class Codes: C07K 2319/31 fusions, other than Fc, for prolonged plasma life, e.g. albumin
• CPC Class Codes: C07K 2319/35 containing a fusion for enhanced stability/folding during expression, e.g. fusions with chaperones or thioredoxin
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins

Abstract

The present invention encompasses albumin fusion proteins. Nucleic acid molecules encoding the albumin fusion proteins of the invention are also encompassed by the invention, as are vectors containing these nucleic acids, host cells transformed with these nucleic acids vectors, and methods of making the albumin fusion proteins of the invention and using these nucleic acids, vectors, and/or host cells. Additionally the present invention encompasses pharmaceutical compositions comprising albumin fusion proteins and methods of treating, preventing, or ameliorating diseases, disorders or conditions using albumin fusion proteins of the invention.

Claims

1. An expression vector comprising a promoter and termination region operably associated with a nucleic acid molecule encoding a leader sequence and an albumin fusion protein comprising two tandemly oriented Glucagon-Like-Peptide 1 (GLP-1) polypeptides fused to albumin or a fragment or variant of albumin, wherein said leader sequence is a modified HSA (human serum albumin)/Kex2 leader sequence having the amino acid sequence of SEQ ID NO:

112.

3. The expression vector of claim 1, wherein said expression vector is constructed from a plasmid selected from:

(a)pSAC35; and

(b)pEE12;

wherein the expression vector comprises an insertion of a nucleic acid molecule encoding a modified HSA (human serum albumin)/Kex2 leader sequence having the amino acid sequence of SEQ ID NO;

1 12 operably associated with an albumin fusion protein comprising two tandemly oriented Glucagon-Like Peptide 1 (GLP-1) polypeptides fused to albumin or a fragment or variant of albumin.

4. The expression vector of claim 3, wherein the expression vector is constructed from a pSAC35 plasmid wherein the expression vector comprises an insertion of a nucleic acid molecule encoding a modified HSA/Kex2 leader sequence having the amino acid sequence of SEQ ID NO:

1 12 operably associated with an albumin fusion protein comprising two tandemly oriented GLP-1 (7-36(A8G)) polypeptides fused to mature albumin.

5. The expression vector of claim 1, wherein said expression vector comprises a nucleic acid sequence encoding the albumin fusion protein of SEQ ID NO:

319.

6. An isolated host cell comprising at least one expression vector of claim 1.

7. The host cell of claim 6, wherein said host cell is a yeast cell or a mammalian cell.

8. The host cell of claim 7, wherein said yeast cell is Saccharomyces Cerevisiae.

9. The host cell of claim 7, wherein said mammalian cell is a CHO (Chinese hamster ovary) cell or a NSO myeloma cell.

10. A method of producing an albumin fusion protein comprising

(a) transforming a host cell comprising at least one expression vector of claim 1;

(b) culturing the host cell in suitable conditions for expression of the albumin fusion protein; and

(c) isolating the albumin fusion protein.

2. The expression vector of claim wherein said promoter is selected from:

(a) a Met25 promoter;

(b) a cbh1 promoter;

(c) a cysD promoter from Aspergillus nidulans;

(d) a modified cbh1 promoter;

(e) an alpha-mating factor promoter; and

(f) a PRB1 promoter.