Fusion polypeptides of human serum albumin and a therapeutically active polypeptide

• US 7,410,779 B2
• Filed: 01/12/2006
• Issued: 08/12/2008
• Est. Priority Date: 01/31/1992
• Status: Expired due to Fees

• Current Assignee: Novozymes Biopharma DK A/S (Novozymes)
• Original Assignee: Novozymes Biopharma Uk Ltd (Novo Nordisk Fonden)
• Inventors: Guitton, Jean-Dominique, Yeh, Patrice, Fleer, Reinhard, Fournier, Alain, Jung, Gérard
• Primary Examiner(s): Landsman; Robert
• Assistant Examiner(s): DANG, IAN D
• Application Number:
  US11/330,353
• Publication Number:
  US 20060105429A1
• Time in Patent Office: 943 Days
• Field of Search: None
• US Class Current: 435/69.1
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 38/21 Interferons [IFN]
• CPC Class Codes: A61K 38/38 Albumins
• CPC Class Codes: A61K 47/642 the peptide or protein in the drug conjugate being a cytokine, e.g. IL2, chemokine, growth factors or interferons being the inactive part of the conjugate
• CPC Class Codes: A61K 47/643 Albumins, e.g. HSA, BSA, ovalbumin or a Keyhole Limpet Hemocyanin [KHL]
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: B65B 69/00 Unpacking of articles or materials, not otherwise provided for
• CPC Class Codes: C07K 14/575 Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin C07K14/665, e.g. corticotropin C07K14/695
• CPC Class Codes: C07K 14/755 Factors VIII , e.g. factor VIII C (AHF), factor VIII Ag (VWF)
• CPC Class Codes: C07K 14/76 Albumins
• CPC Class Codes: C07K 14/765 Serum albumin, e.g. HSA
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/02 containing a signal sequence
• CPC Class Codes: C07K 2319/31 fusions, other than Fc, for prolonged plasma life, e.g. albumin
• CPC Class Codes: C07K 2319/33 fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
• CPC Class Codes: C12N 9/6456 Plasminogen activators

Abstract

The invention includes biologically active polypeptides comprising a therapeutically active polypeptide fused to human serum albumin or a variant of human serum albumin, methods for the preparation of such fusion polypeptides, nucleotide sequences encoding such fusion polypeptides, expression cassettes comprising such nucleotide sequences, self-replicating plasmids containing such expression cassettes, and pharmaceutical compositions containing such fusion polypeptides.

Claims

1. A nucleic acid molecule comprising a polynucleotide encoding a fusion protein wherein said fusion protein comprises a hormone and a mature form of albumin, wherein (i) said fusion protein has a higher plasma stability than the unfused hormone, (ii) said fusion protein retains the therapeutic activity of the unfused hormone, (iii) and said mature form of albumin is located either at the N-terminus or C-terminus of said fusion protein.

2. The nucleic acid molecule of claim 1, wherein said fusion protein comprises an N-terminal methionine.

3. The nucleic acid molecule of claim 1, wherein said fusion protein comprises a peptide linker.

4. The nucleic acid molecule of claim 1, wherein said fusion protein comprises a secretion signal sequence.

5. The nucleic acid molecule of claim 1, wherein said hormone is fused to the N-terminal end of said mature form of albumin.

6. The nucleic acid molecule of claim 1, wherein said hormone is fused to the C-terminal end of said mature form of albumin.

7. The nucleic acid molecule of claim 1, which further comprises a heterologous polynucleotide.

8. The nucleic acid molecule of claim 7, wherein said heterologous polynucleotide is a vector sequence.

9. The nucleic acid molecule of claim 7, wherein said heterologous polynucleotide is a promoter sequence.

10. The nucleic acid molecule of claim 9, wherein said promoter sequence is a promoter selected from:

(a) a hybrid promoter;

(b) a constitutive promoter;

(c) a regulatable promoter;

(d) a yeast phosphoglycerate kinase (PGK) promoter;

(e) a yeast glyceraldehyde-3-phosphate dehydrogenase (GDP) promoter;

(f) a yeast lactase (LAC4) promoter;

(g) a yeast enolase (ENO) promoter;

(h) a yeast alcohol dehydrogenase (ADH) promoter;

(i) a yeast phosphatase (PHO5) promoter;

(j) a lambda bacteriophage P_L promoter;

(k) a lambda bacteriophage P_R promoter;

(l) a tryptophan P_trp promoter; and

(m) a lactose P_lac promoter.

11. The nucleic acid molecule of claim 7, wherein said heterologous polynucleotide is a selectable marker.

12. The nucleic acid molecule of claim 11, wherein said selectable marker is a selectable marker selected from:

(a) the URA3 gene;

(b) geneticin resistance;

(c) metal iron resistance; and

(d) ampicillin resistance.

13. The nucleic acid molecule of claim 7, wherein said heterologous polynucleotide is a region for termination of transcription.

14. The nucleic acid molecule of claim 1, wherein said nucleic acid molecule is part of an expression cassette.

15. A vector comprising the nucleic acid molecule of claim 1.

16. An isolated host cell comprising the nucleic acid molecule of claim 1.

17. An isolated host cell comprising the vector of claim 15.

18. The isolated host cell of claim 16 or 17, wherein said isolated host cell is prokaryotic or eukaryotic.

19. The isolated host cell of claim 18, wherein said isolated host cell is a yeast cell.

20. The isolated host cell of claim 19, wherein said yeast cell is S. cerevisiae.

21. The isolated host cell of claim 18, wherein said isolated host cell is an animal cell.

22. The isolated host cell of claim 21, wherein said animal cell is a CHO cell or a COS cell.

23. The isolated host cell of claim 18, wherein said isolated host cell is a fungi cell.

24. A method for producing a fusion protein comprising:

(a) culturing the isolated host cell of claim 16 under conditions suitable to produce the fusion protein encoded by said polynucleotide; and

(b) recovering said fusion protein.

25. A fusion protein produced by the method of claim 24.