Albumin fusion proteins

• US 20070027306A1
• Filed: 08/08/2006
• Published: 02/01/2007
• Est. Priority Date: 02/09/2004
• Status: Active Grant

• Current Assignee: Human Genome Sciences Incorporated (GSK plc)
• Original Assignee: Human Genome Sciences Incorporated (GSK plc)
• Inventors: Rosen, Craig, Haseltine, William, Ruben, Steven
• Granted Patent: US 7,569,384 B2
• Time in Patent Office: Days
• US Class Current: 530/399
• 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 to a nucleic acid molecule encoding a leader sequence and an albumin fusion protein comprising two tandemly oriented GLP-1 polypeptides fused to albumin or a fragment or variant of albumin.

2. The vector of claim 1, wherein said leader sequence is modified to produce better expression yield and/or better cleavage yield of said albumin fusion protein than an unmodified leader sequence.

3. The vector of claim 2, wherein said leader sequence is a modified HSA/kex2 signal sequence.

4. The vector of claim 1, wherein said two tandemly oriented GLP-1 polypeptides are GLP-1(7-36(A8G)) (SEQ ID NO:

387).

5. The vector of claim 1, wherein said leader sequence is selected from the group consisting of:

(a) a pre-pro region of the HSA leader sequence;

(b) a killer toxin leader sequence;

(c) an HSA/Kex2 leader sequence;

(d) a yeast mating factor alpha signal sequence;

(e) a modified HSA/kex2 leader sequence;

(f) an MPIF-1 leader sequence; and

(g) a consensus leader sequence.

6. The vector of claim 5, wherein said leader sequence is a modified HSA-Kex2 leader sequence encoding the amino acids of SEQ ID NO:

112.

7. The vector of claim 1, wherein said promoter is selected from the group consisting of:

(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.

8. The vector of claim 1, wherein said expression vector is selected from the group consisting of:

(a) pSAC35;

(b) pEE12; and

(c) pC4;

HSA.

10. A host cell comprising at least one vector of claim 1.

11. The host cell of claim 10, wherein said host cell is a yeast cell or a mammalian cell.

12. The host cell of claim 11, wherein said yeast cell is Saccharomyces cerevisiae.

13. The host cell of claim 11, wherein said mammalian cell is CHO or NSO.

14. A method of producing an albumin fusion protein comprising

(a) transforming a host cell comprising at least one 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.

15. The albumin fusion protein produced by the method of claim 14.

16. The albumin fusion protein of claim 15, wherein said albumin fusion protein is expressed from a host cell containing an albumin fusion construct selected from the group consisting of:

(a) Construct ID 3610; and

(b) Construct ID 3696.

18. A composition comprising the albumin fusion protein of claim 15 and a pharmaceutically acceptable carrier.

19. A method of treating a patient for a disease, condition, or disorder, comprising administering an effective amount of the albumin fusion protein of claim 15, wherein said disease, condition, or disorder is selected from hyperglycemia, diabetes, diabetes insipidus, insulin resistance, insulin deficiency, hyperlipidemia, hyperketonemia, obesity, heart disease, infections, retinopathy, ulcers, metabolic disorders, immune disorders, vascular disorders, suppression of body weight, suppression of appetite, syndrome X, cognitive impairment, and memory loss.

20. A method of stimulating insulin synthesis and release, enhancing adipose, muscle or liver tissue sensitivity towards insulin uptake, stimulating glucose uptake, slowing digestive process, or blocking the secretion of glucagon in a patient, comprising administering to said patient an effective amount of the albumin fusion protein of claim 15.

9. An pSAC35 expression-vector comprising a nucleic acid molecule encoding a modified HSA/kex2 leader sequence and an albumin fusion protein comprising two tandemly oriented GLP-1(7-36(A8G)) fused to mature albumin.

17. An albumin fusion protein produced from a host cell comprising Construct ID 3610, wherein said albumin fusion protein comprises amino acids 25 to 669 of SEQ ID NO:

319.

21. A method of treating a patient for a disease, condition, or disorder, comprising administering an effective amount of the albumin fusion protein of claim 17, wherein said disease, condition, or disorder is selected from hyperglycemia, diabetes, diabetes insipidus, insulin resistance, insulin deficiency, hyperlipidemia, hyperketonemia, obesity, heart disease, infections, retinopathy, ulcers, metabolic disorders, immune disorders, vascular disorders, suppression of body weight, suppression of appetite, syndrome X, cognitive impairment, and memory loss.

22. A method of stimulating insulin synthesis and release, enhancing adipose, muscle or liver tissue sensitivity towards insulin uptake, stimulating glucose uptake, slowing digestive process, or blocking the secretion of glucagon in a patient, comprising administering to said patient an effective amount of the albumin fusion protein of claim 17.