Albumin fusion proteins

• US 20070048282A1
• Filed: 08/08/2006
• Published: 03/01/2007
• Est. Priority Date: 02/09/2004
• Status: Abandoned Application

• Current Assignee: Human Genome Sciences Incorporated (GlaxoSmithKline PLC)
• Original Assignee: Human Genome Sciences Incorporated (GlaxoSmithKline PLC)
• Inventors: Bell, Adam, Shi, Yanggu, Moore, Paul, Rosen, Craig, LaFleur, David, Haseltine, William, Bock, Jason
• Application Number:
  US11/500,314
• Publication Number:
  US 20070048282A1
• Time in Patent Office: Days
• US Class Current: 424/85.700
• 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 albumin fusion protein comprising a member selected from the group consisting of:

(a) a Therapeutic protein;

X and albumin comprising the amino acid sequence of SEQ ID NO;

1;

(b) a Therapeutic protein;

X and a fragment or a variant of the amino acid sequence of SEQ ID NO;

1, wherein said fragment or variant has albumin activity;

(c) a Therapeutic protein;

X and a fragment or a variant of the amino acid sequence of SEQ ID NO;

1, wherein said fragment or variant has albumin activity, and further wherein said albumin activity is the ability to prolong the shelf life of the Therapeutic protein;

X compared to the shelf-life of the Therapeutic protein;

X in an unfused state;

(d) a Therapeutic protein;

X and a fragment or a variant of the amino acid sequence of SEQ ID NO;

1, wherein said fragment or variant has albumin activity, and further wherein the fragment or variant comprises the amino acid sequence of amino acids 1-387 of SEQ ID NO;

1;

(e) a fragment or variant of a Therapeutic protein;

X and albumin comprising the amino acid sequence of SEQ ID NO;

1, wherein said fragment or variant has a biological activity of the Therapeutic protein;

X;

(f) a Therapeutic protein;

X, or fragment or variant thereof, and albumin, or fragment or variant thereof, of (a) to (e), wherein the Therapeutic protein;

X, or fragment or variant thereof, is fused to the N-terminus of albumin, or the N-terminus of the fragment or variant of albumin;

(g) a Therapeutic protein;

X, or fragment or variant thereof, and albumin, or fragment or variant thereof, of (a) to (e), wherein the Therapeutic protein;

X, or fragment or variant thereof, is fused to the C-terminus of albumin, or the C-terminus of the fragment or variant of albumin;

(h) a Therapeutic protein;

X, or fragment or variant thereof, and albumin, or fragment or variant thereof, of (a) to (e), wherein the Therapeutic protein;

X, or fragment or variant thereof, is fused to the N- terminus and C-terminus of albumin, or the N-terminus and the C-terminus of the fragment or variant of albumin;

(i) a Therapeutic protein;

X, or fragment or variant thereof, and albumin, or fragment or variant thereof, of (a) to (e), which comprises a first Therapeutic protein;

X, or fragment or variant thereof, and a second Therapeutic protein;

X, or fragment or variant thereof, wherein said first Therapeutic protein;

X, or fragment or variant thereof, is different from said second Therapeutic protein;

X, or fragment or variant thereof;

(j) a Therapeutic protein;

X, or fragment or variant thereof, and albumin, or fragment or variant thereof, of (a) to (i), wherein the Therapeutic protein;

X, or fragment or variant thereof, is separated from the albumin or the fragment or variant of albumin by a linker; and

(k) a Therapeutic protein;

X, or fragment or variant thereof, and albumin, or fragment or variant thereof, of (a) to (j), wherein the albumin fusion protein has the following formula;

R1-L-R2;

R2-L-R1;

or R1-L-R2-L-R1,

wherein R1 is Therapeutic protein;

X, or fragment or variant thereof, L is a peptide linker, and R2 is albumin comprising the amino acid sequence of SEQ ID NO;

1 or a fragment or variant of albumin;

wherein said Therapeutic protein;

X is a human growth hormone, an interferon alpha, an interferon beta, a natriuretic peptide, a lysosomal glucocerebrosidase, a brain-derived neurotrophic factor isoform, a glucagon-like peptide 1, an arginine deiminase, a uricase, an interferon hybrid, an interleukin-2, an adrenomedullin, a tweak Receptor, a Kiss-1, an ephrin B1, or a B7-H3.

2. The albumin fusion protein of claim 1, wherein the shelf-life or plasma stability of the albumin fusion protein is greater than the shelf-life or plasma stability of the Therapeutic protein:

X, or fragment or variant thereof, in an unfused state.

3. The albumin fusion protein of claim 1, wherein the in vitro biological activity of the Therapeutic protein:

X, or fragment or variant thereof, fused to albumin, or fragment or variant thereof, is greater than the in vitro biological activity of the Therapeutic protein;

X, or fragment or variant thereof, in an unfused state.

4. The albumin fusion protein of claim 1, wherein the in vivo biological activity of the Therapeutic protein:

X, or fragment or variant thereof, fused to albumin, or fragment or variant thereof, is greater than the in vivo biological activity of the Therapeutic protein;

X, or fragment or variant thereof, in an unfused state.

5. The albumin fusion protein of claim 1, which is non-glycosylated.

6. The albumin fusion protein of claim 1, which is expressed in yeast.

7. The albumin fusion protein of claim 6, wherein the yeast is glycosylation deficient or glycosylation and protease deficient.

8. The albumin fusion protein of claim 1, which is expressed by a mammalian cell.

9. The albumin fusion protein of claim 1, which further comprises a secretion leader sequence.

10. A composition comprising the albumin fusion protein of claim 1 and a pharmaceutically acceptable carrier.

11. A kit comprising the composition of claim 10.

12. A method of treating a disease or disorder in a patient, comprising the step of administering the albumin fusion protein of claim 1.

13. The method of claim 12, wherein the disease or disorder comprises indication:

Y.

14. A method of treating a patient with a disease or disorder that is modulated by Therapeutic protein:

X, or fragment or variant thereof, comprising the step of administering an effective amount of the albumin fusion protein of claim 1.

15. The method of claim 14, wherein the disease or disorder is indication:

Y.

17. A nucleic acid molecule comprising a polynucleotide sequence encoding the albumin fusion protein of claim 1.

18. An albumin fusion construct comprising the nucleic acid molecule of claim 17.

19. A vector comprising the nucleic acid molecule of claim 17.

20. A host cell comprising the albumin fusion construct of claim 18.

21. A method of producing an albumin fusion protein comprising:

(a) culturing the host cell of claim 20 under conditions suitable for expression of the albumin fusion protein; and

(b) isolating the albumin fusion protein.

22. The albumin fusion protein produced by the method of claim 21.

16. A method of extending the shelf life or plasma stability of Therapeutic protein:

X, or fragment or variant thereof, comprising the step of fusing the Therapeutic protein;

X, or fragment or variant thereof, to albumin, or fragment or variant thereof, sufficient to extend the shelf-life or plasma stability of the Therapeutic protein;

X, or fragment or variant thereof, compared to the shelf-life of the Therapeutic protein;

X, or fragment or variant thereof, in an unfused state.