Albumin fusion proteins

• US 20050054051A1
• Filed: 08/20/2004
• Published: 03/10/2005
• Est. Priority Date: 04/12/2001
• Status: Abandoned Application

• Current Assignee: Human Genome Sciences Incorporated (GSK plc)
• Original Assignee: Human Genome Sciences Incorporated (GSK plc)
• Inventors: Rosen, Craig A., Haseltine, William A.
• Application Number:
  US10/922,142
• Publication Number:
  US 20050054051A1
• Time in Patent Office: Days
• US Class Current: 435/69.700
• CPC Class Codes: A61K 2039/54 characterised by the route of administration
• 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: C07K 14/61 Growth hormone [GH], i.e. somatotropin
• CPC Class Codes: C07K 14/765 Serum albumin, e.g. HSA
• CPC Class Codes: C07K 2319/01 containing a localisation/targetting motif
• CPC Class Codes: C07K 2319/21 containing a His-tag
• CPC Class Codes: C07K 2319/31 fusions, other than Fc, for prolonged plasma life, e.g. albumin
• CPC Class Codes: C07K 2319/50 containing protease site
• CPC Class Codes: C07K 2319/75 containing a fusion for activation of a cell surface receptor, e.g. thrombopoeitin, NPY and other peptide hormones
• 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, disordrs or conditions using albumin fusion proteins of the invention.

Claims

1. An albumin fusion protein comprising a member selected from:

(a) a Therapeutic protein X and albumin comprising the amino acid sequence of SEQ ID NO;

18;

(b) a Therapeutic protein X and a fragment or a variant of the amino acid sequence of SEQ ID NO;

18, 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;

18, 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;

18, wherein said fragment or variant has albumin activity, and further wherein said albumin activity is the ability to increase the half-life of the Therapeutic protein X compared to the half-life of the Therapeutic protein X in an unfused state;

(e) a Therapeutic protein X and a fragment or a variant of the amino acid sequence of SEQ ID NO;

18, 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;

18;

(f) a fragment or variant of a Therapeutic protein X and albumin comprising the amino acid sequence of SEQ ID NO;

18, wherein said fragment or variant has a biological activity of the Therapeutic Protein X;

(g) a Therapeutic protein X, or fragment or variant thereof, and albumin, or fragment or variant thereof, of (a) to (f), 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;

(h) a Therapeutic protein X, or fragment or variant thereof, and albumin, or fragment or variant thereof, of (a) to (f), 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;

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

(k) 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

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

R1-L-R2;

R2-L-R1;

or R1-L-R2-L-R1,

 

and further 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;

18 or a fragment or variant of albumin.

2. The albumin fusion protein of claim 1, wherein the shelf-life of the albumin fusion protein is greater than the shelf-life 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.

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

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

15. The albumin fusion protein of claim 14, wherein the yeast is glycosylation deficient.

16. The albumin fusion protein of claim 14 wherein the yeast is glycosylation and protease deficient.

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

18. The albumin fusion protein of claim 1, wherein the albumin fusion protein is expressed by a mammalian cell in culture.

19. The albumin fusion protein of claim 1, wherein the albumin fusion protein further comprises a secretion leader sequence.

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

21. A kit comprising the composition of claim 20.

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

23. The method of claim 22, wherein the disease or disorder comprises indication Y.

24. 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.

25. The method of claim 24, wherein the disease or disorder is indication Y.

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

28. A vector comprising the nucleic acid molecule of claim 27.

29. A host cell comprising the nucleic acid molecule of claim 28.

30. The albumin fusion protein of claim 1, wherein the half-life of the albumin fusion protein is greater than the half-life of the Therapeutic protein X, or fragment or variant thereof, in an unfused state.

5. An albumin fusion protein comprising a Therapeutic protein X, or fragment or variant thereof, inserted into an albumin, or fragment or variant thereof, comprising the amino acid sequence of SEQ ID NO:

18 or fragment or variant thereof.

7. The albumin fusion protein of claim 5, wherein said albumin fusion protein comprises a portion of albumin sufficient to prolong the shelf-life of the Therapeutic protein X, or fragment or variant thereof, as compared to the shelf-life of the Therapeutic protein X, or fragment or variant thereof, in an unfused state.

9. The albumin fusion protein of claim 5, wherein said albumin fusion protein comprises a portion of albumin sufficient to prolong the in vitro biological activity of the Therapeutic protein X, or fragment or variant thereof, fused to albumin as compared to the in vitro biological activity of the Therapeutic protein X, or fragment or variant thereof, in an unfused state.

11. The albumin fusion protein of claim 5 wherein said albumin fusion protein comprises a portion of albumin sufficient to prolong the in vivo biological activity of the Therapeutic protein X, or fragment or variant thereof, fused to albumin compared to the in vivo biological activity of the Therapeutic protein X, or fragment or variant thereof, in an unfused state.

31. The albumin fusion protein of claim 5, wherein said albumin fusion protein comprises a portion of albumin sufficient to increase the half-life of the Therapeutic protein X, or fragment or variant thereof, as compared to the half-life of the Therapeutic protein X, or fragment or variant thereof, in an unfused state.

6. An albumin fusion protein comprising a Therapeutic protein X, or fragment or variant thereof, inserted into an albumin, or fragment or variant thereof, comprising an amino acid sequence selected from:

(a) amino acids 54 to 61 of SEQ ID NO;

18;

(b) amino acids 76 to 89 of SEQ ID NO;

18;

(c) amino acids 92 to 100 of SEQ ID NO;

18;

(d) amino acids 170 to 176 of SEQ ID NO;

18;

(e) amino acids 247 to 252 of SEQ ID NO;

18;

(f) amino acids 266 to 277 of SEQ ID NO;

18;

(g) amino acids 280 to 288 of SEQ ID NO;

18;

(h) amino acids 362 to 368 of SEQ ID NO;

18;

(i) amino acids 439 to 447 of SEQ ID NO;

18;

(j) amino acids 462 to 475 of SEQ ID NO;

18;

(k) amino acids 478 to 486 of SEQ ID NO;

18; and

(l) amino acids 560 to 566 of SEQ ID NO;

18.

8. The albumin fusion protein of claim 6, wherein said albumin fusion protein comprises a portion of albumin sufficient to prolong the shelf-life of the Therapeutic protein X, or fragment or variant thereof, as compared to the shelf-life of the Therapeutic protein X, or fragment or variant thereof, in an unfused state.

10. The albumin fusion protein of claim 6, wherein said albumin fusion protein comprises a portion of albumin sufficient to prolong the in vitro biological activity of the Therapeutic protein X, or fragment or variant thereof, fused to albumin as compared to the in vitro biological activity of the Therapeutic protein X, or fragment or variant thereof, in an unfused state.

12. The albumin fusion protein of claim 6 wherein said albumin fusion protein comprises a portion of albumin sufficient to prolong the in vivo biological activity of the Therapeutic protein X, or fragment or variant thereof, fused to albumin compared to the in vivo biological activity of the Therapeutic protein X, or fragment or variant thereof, in an unfused state.

32. The albumin fusion protein of claim 6, wherein said albumin fusion protein comprises a portion of albumin sufficient to increase the half-life of the Therapeutic protein X, or fragment or variant thereof, as compared to the half-life of the Therapeutic protein X, or fragment or variant thereof, in an unfused state.

26. A method of extending the shelf life 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 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.

33. A method of increasing the half-life 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 increase the half-life of the Therapeutic protein X, or fragment or variant thereof, compared to the half-life of the Therapeutic protein X, or fragment or variant thereof, in an unfused state.