Albumin fusion proteins

• US 6,926,898 B2
• Filed: 04/12/2001
• Issued: 08/09/2005
• Est. Priority Date: 04/12/2000
• Status: Expired due to Term

• Current Assignee: Human Genome Sciences Incorporated (GSK plc)
• Original Assignee: Human Genome Sciences Incorporated (GSK plc)
• Inventors: Rosen, Craig A., Haseltine, William A.
• Primary Examiner(s): Weber, Jon
• Assistant Examiner(s): Robinson, Hope A.
• Application Number:
  US09/832,929
• Publication Number:
  US 20040171123A1
• Time in Patent Office: 1,580 Days
• Field of Search: 424/192.1, 435/7.1, 435/6, 435/320.1, 530/350, 530/300, 530/387.1, 536/23.1, 514/2, 514/12
• US Class Current: 424/192.1
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A61K 2039/54 characterised by the route of administration
• CPC Class Codes: A61K 38/21 Interferons [IFN]
• CPC Class Codes: A61K 38/212 IFN-alpha
• CPC Class Codes: A61K 38/38 Albumins
• CPC Class Codes: A61K 38/4846 Factor VII (3.4.21.21); Factor IX (3.4.21.22); Factor Xa (3.4.21.6); Factor XI (3.4.21.27); Factor XII (3.4.21.38)
• CPC Class Codes: A61K 47/42 Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein oligopeptides having up to five amino acids A61K47/183; polyamino acids A61K47/34
• CPC Class Codes: A61K 47/65 Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 13/00 Drugs for disorders of the urinary system
• CPC Class Codes: A61P 13/02 of urine or of the urinary tract, e.g. urine acidifiers
• CPC Class Codes: A61P 13/08 of the prostate
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 15/00 : Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 15/08 : for gonadal disorders or for enhancing fertility, e.g. inducers of ovulation or of spermatogenesis
• CPC Class Codes: A61P 15/10 : for impotence
• CPC Class Codes: A61P 15/18 : Feminine contraceptives
• CPC Class Codes: A61P 17/00 : Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 : for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 17/06 : Antipsoriatics
• CPC Class Codes: A61P 17/12 : Keratolytics, e.g. wart or anti-corn preparations
• CPC Class Codes: A61P 17/14 : for baldness or alopecia
• CPC Class Codes: A61P 19/00 : Drugs for skeletal disorders
• CPC Class Codes: A61P 19/02 : for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/08 : for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 19/10 : for osteoporosis
• CPC Class Codes: A61P 21/00 : Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 21/04 : for myasthenia gravis
• CPC Class Codes: A61P 25/00 : Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/02 : for peripheral neuropathies
• CPC Class Codes: A61P 25/08 : Antiepileptics; Anticonvulsants
• CPC Class Codes: A61P 25/16 : Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 : for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 27/02 : Ophthalmic agents
• CPC Class Codes: A61P 29/00 : Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/10 : for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 3/14 : for calcium homeostasis vitamin D A61P3/02; parathyroid hormones A61P5/18; calcitonin A61P5/22; osteoporosis A61P19/10; bone metastasis A61P35/04
• CPC Class Codes: A61P 31/00 : Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/12 : Antivirals
• CPC Class Codes: A61P 31/14 : for RNA viruses
• CPC Class Codes: A61P 31/16 : for influenza or rhinoviruses
• CPC Class Codes: A61P 31/18 : for HIV
• CPC Class Codes: A61P 31/20 : for DNA viruses
• CPC Class Codes: A61P 31/22 : for herpes viruses
• CPC Class Codes: A61P 33/02 : Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
• CPC Class Codes: A61P 33/06 : Antimalarials
• CPC Class Codes: A61P 33/12 : Schistosomicides
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 35/02 : specific for leukemia
• CPC Class Codes: A61P 35/04 : specific for metastasis
• CPC Class Codes: A61P 37/00 : Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 : Immunomodulators
• CPC Class Codes: A61P 37/04 : Immunostimulants
• 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 39/02 : Antidotes
• CPC Class Codes: A61P 41/00 : Drugs used in surgical methods, e.g. surgery adjuvants for preventing adhesion or for vitreum substitution
• 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: A61P 5/10 : of the posterior pituitary hormones, e.g. oxytocin, ADH
• CPC Class Codes: A61P 5/14 : of the thyroid hormones, e.g. T3, T4
• CPC Class Codes: A61P 5/40 : Mineralocorticosteroids, e.g. aldosterone; Drugs increasing or potentiating the activity of mineralocorticosteroids
• CPC Class Codes: A61P 7/00 : Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/04 : Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
• CPC Class Codes: A61P 7/06 : Antianaemics
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/06 : Antiarrhythmics
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61P 9/12 : Antihypertensives
• CPC Class Codes: C07K 14/56 : IFN-alpha
• CPC Class Codes: C07K 14/61 : Growth hormone [GH], i.e. somatotropin
• CPC Class Codes: C07K 14/62 : Insulins
• CPC Class Codes: C07K 14/65 : Insulin-like growth factors, i.e. somatomedins, e.g. IGF-1, IGF-2
• CPC Class Codes: C07K 14/705 : Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 14/7151 : for tumor necrosis factor [TNF], for lymphotoxin [LT]
• 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 2319/21 : 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
• CPC Class Codes: C12N 9/96 : Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
• CPC Class Codes: Y02A 50/30 : Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

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 the group consisting of:

(a) an antibody or antibody fragment that specifically binds human epidermal growth factor receptor-2 (HER2) and albumin, wherein albumin comprises the amino acid sequence of SEQ ID NO;

18;

(b) an antibody or antibody fragment that specifically binds human epidermal growth factor receptor-2 (HER2) and a fragment of the amino acid sequence of SEQ ID NO;

18, wherein said fragment of the amino acid sequence of SEQ ID NO;

18 has the ability to prolong the shelf-life of the antibody or antibody fragment that specifically binds HER2, compared to the shelf-life of the antibody or antibody fragment that specifically binds HER2, in an unfused state;

(c) an antibody or antibody fragment that specifically binds human epidermal growth factor receptor-2 (HER2) and a fragment of the amino acid sequence of SEQ ID NO;

18, wherein said fragment has the ability to prolong the shelf-life of the antibody or antibody fragment that specifically binds HER2 compared to the shelf-life of the antibody or antibody fragment that specifically binds HER2, in an unfused state, and further wherein the said fragment comprises amino acid residues 1-387 of SEQ ID NO;

18;

(d) a fragment of an antibody that specifically binds human epidermal growth factor receptor-2 (HER2) and albumin comprising the amino acid sequence of SEQ ID NO;

18, wherein said fragment has a biological activity of the antibody that specifically binds HER2;

(e) an antibody or antibody fragment that specifically binds human epidermal growth factor receptor-2 (HER2), and albumin, or fragment thereof, of (a) to (d), wherein the antibody or antibody fragment that specifically binds HER2, is fused to the N-terminus of albumin or the N-terminus of the fragment of albumin;

(f) an antibody or antibody fragment that specifically binds human epidermal growth factor receptor-2 (HER2), and albumin, or fragment thereof, of (a) to (d), wherein the antibody or antibody fragment that specifically binds HER2, is fused to the C-terminus of albumin, or the C-terminus of the fragment of albumin;

(g) an antibody or antibody fragment that specifically binds human epidermal growth factor receptor-2 (HER2), and albumin or fragment thereof, of (a) to (d), wherein the an antibody or antibody fragment that specifically binds HER2, is fused to the N-terminus and C-terminus of albumin, or the N-terminus and the C-terminus of the fragment of albumin;

(h) an antibody or antibody fragment that specifically binds human epidermal growth factor receptor-2 (HER2), and albumin or fragment thereof, of (a) to (g), wherein the an antibody or antibody fragment that specifically binds HER2, is separated from the albumin or the fragment of albumin by a linker; and

(i) an antibody or antibody fragment that specifically binds human epidermal growth factor receptor-2 (HER2), and albumin or fragment thereof, of (a) to (h), 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 an antibody or antibody fragment that specifically binds HER2, L is a peptide linker, and R2 is albumin comprising the amino acid sequence of SEQ ID NO;

18 or a fragment 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 antibody or antibody fragment that specifically binds HER2, in an unfused state.

3. The albumin fusion protein of claim 1, wherein the in vitro biological activity of the antibody or antibody fragment that specifically binds HER2, fused to albumin, or fragment thereof, is greater than the in vitro biological activity of the antibody or antibody fragment that specifically binds HER2, in an unfused state.

4. The albumin fusion protein of claim 1, wherein the in vivo biological activity of the antibody or antibody fragment that specifically binds HER2, fused to albumin, or fragment thereof, is greater than the in vivo biological activity of the antibody or antibody fragment that specifically binds HER2, in an unfused state.

13. The albumin fusion protein of any one of claims 1-12, which is non-glycosylated.

14. The albumin fusion protein of any one of claims 1-12, 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 any one of claims 1-12, which is expressed by a mammalian cell.

18. The albumin fusion protein of any one of claims 1-12, wherein the albumin fusion protein is expressed by a mammalian cell in culture.

19. The albumin fusion protein of any one of claims 1-12, wherein the albumin fusion protein further comprises a secretion leader sequence.

20. A composition comprising the albumin fusion protein of any one of claims 1-12 and a pharmaceutically acceptable carrier.

21. A kit comprising the composition of claim 20.

23. A nucleic acid molecule comprising a polynucleotide sequence encoding the albumin fusion protein of any one of claims 1-12.

24. A vector comprising, the nucleic acid molecule of claim 23.

25. A host cell comprising the nucleic acid molecule of claim 24.

5. An albumin fusion protein comprising an antibody or antibody fragment that specifically binds HER2, inserted into an albumin, or fragment thereof, comprising the amino acid sequence of SEQ ID NO:

18 or fragment thereof.

7. The albumin fusion protein of claim 5, wherein said albumin fusion protein comprises a fragment of albumin sufficient to prolong the shelf-life of the antibody or antibody fragment that specifically binds HER2, as compared to the shelf-life of the antibody or antibody fragment that specifically binds HER2, in an unfused state.

9. The albumin fusion protein of claim 5, wherein said albumin fusion protein comprises a fragment of albumin sufficient to prolong the in vitro biological activity of the antibody or antibody fragment that specifically binds HER2, fused to albumin as compared to the in vitro biological activity of the antibody or antibody fragment that specifically binds HER2, in an unfused state.

11. The albumin fusion protein of claim 5, wherein said albumin fusion protein comprises a fragment of albumin sufficient to prolong the in vivo biological activity of the antibody or antibody fragment that specifically binds HER2, fused to albumin as compared to the in vivo biological activity of the antibody or antibody fragment that specifically binds HER2, in an unfused state.

6. An albumin fusion protein comprising an antibody or antibody fragment that specifically binds human epidermal growth factor receptor-2 (HER2), inserted into an albumin, or fragment thereof, comprising an amino acid sequence selected from the group consisting of:

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

18;

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

18;

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

18;

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

18;

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

18;

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

18;

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

18;

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

18;

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

18;

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

18;

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

18; and

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

18.

8. The albumin fusion protein of claim 6, wherein said albumin fusion protein comprises a fragment of albumin sufficient to prolong the shelf-life of the antibody or antibody fragment that specifically binds HER2, as compared to the shelf-life of the antibody or antibody fragment that specifically binds HER2, in an unfused state.

10. The albumin fusion protein of claim 6, wherein said albumin fusion protein comprises a fragment of albumin sufficient to prolong the in vitro biological activity of the antibody or antibody fragment that specifically binds HER2, fused to albumin as compared to the in vitro biological activity of the antibody or antibody fragment that specifically binds HER2, in an unfused state.

12. The albumin fusion protein of claim 6, wherein said albumin fusion protein comprises a fragment of albumin sufficient to prolong the in vivo biological activity of the antibody or antibody fragment that specifically binds HER2, fused to albumin as compared to the in vivo biological activity of the antibody or antibody fragment that specifically binds HER2, in an unfused state.

22. A method of extending the shelf-life of an antibody that specifically binds human epidermal growth factor receptor-2 (HER2) or fragment thereof, comprising the step of fusing the antibody that specifically binds HER2 or fragment thereof, to albumin or albumin fragment, sufficient to extend the shelf-life of the antibody that specifically binds HER2 or fragment thereof, compared to the shelf-life of the antibody that specifically binds HER2 or fragment thereof, in an unfused state.