Serpin fusion polypeptides and methods of use thereof

• US 8,980,266 B2
• Filed: 06/28/2012
• Issued: 03/17/2015
• Est. Priority Date: 06/28/2011
• Status: Active Grant

• Current Assignee: Inhibrx, Inc.
• Original Assignee: InhibRx, LLC (Inhibrx, Inc.)
• Inventors: Eckelman, Brendan, Timmer, John, Nguy, Peter L., Guenther, Grant B., Deveraux, Quinn
• Primary Examiner(s): STANFIELD, CHERIE MICHELLE
• Application Number:
  US13/536,976
• Publication Number:
  US 20130011398A1
• Time in Patent Office: 992 Days
• Field of Search: 530/350, 530/387.3, 530/391.1, 424/178.1
• US Class Current: 424/134.1
• 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 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 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• 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 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• 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 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/10 Antimycotics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 : Immunomodulators
• CPC Class Codes: A61P 37/06 : Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 5/00 : Drugs for disorders of the endocrine system
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• 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: C07K 14/525 : Tumour necrosis factor [TNF]
• CPC Class Codes: C07K 14/7151 : for tumor necrosis factor [TNF], for lymphotoxin [LT]
• CPC Class Codes: C07K 14/7155 : for interleukins [IL]
• CPC Class Codes: C07K 14/76 : Albumins
• CPC Class Codes: C07K 14/765 : Serum albumin, e.g. HSA
• CPC Class Codes: C07K 14/811 : Serine protease (E.C. 3.4.21) inhibitors
• CPC Class Codes: C07K 14/8121 : Serpins
• CPC Class Codes: C07K 14/8125 : Alpha-1-antitrypsin
• CPC Class Codes: C07K 16/241 : Tumor Necrosis Factors
• CPC Class Codes: C07K 16/40 : against enzymes
• CPC Class Codes: C07K 2317/52 : Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/522 : CH1 domain
• CPC Class Codes: C07K 2317/524 : CH2 domain
• CPC Class Codes: C07K 2317/526 : CH3 domain
• CPC Class Codes: C07K 2317/53 : Hinge
• CPC Class Codes: C07K 2317/56 : variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide
• CPC Class Codes: C07K 2319/30 : Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C07K 2319/31 : fusions, other than Fc, for prolonged plasma life, e.g. albumin
• CPC Class Codes: C07K 2319/70 : containing domain for protein-protein interaction

Abstract

This invention relates to molecules, particularly polypeptides, more particularly fusion proteins that include a serpin polypeptide or an amino acid sequence that is derived from a serpin and second polypeptide comprising of at least one the following: an Fc polypeptide or an amino acid sequence that is derived from an Fc polypeptide; a cytokine targeting polypeptide or a sequence derived from a cytokine targeting polypeptide; a WAP domain containing polypeptide or a sequence derived from a WAP containing polypeptide; and an albumin polypeptide or an amino acid sequence that is derived from a serum albumin polypeptide. This invention also relates to methods of using such molecules in a variety of therapeutic and diagnostic indications, as well as methods of producing such molecules.

Claims

1. An isolated fusion protein comprising at least one human serpin polypeptide comprising a human alpha-1 antitrypsin (AAT) polypeptide comprising the amino acid sequence of SEQ ID NO:

1 operably linked to

an immunoglobulin Fc polypeptide comprising the amino acid sequence of SEQ ID NO;

6.

2. The isolated fusion protein of claim 1, wherein AAT polypeptide comprises the amino acid sequence of SEQ ID NO:

2.

3. The isolated fusion protein of claim 1, wherein the serpin polypeptide and the immunoglobulin Fc polypeptide are operably linked via a hinge region, a linker region, or both a hinge region and linker region.

4. The isolated fusion protein of claim 3, wherein the hinge region, the linker region or both the hinge region and the linker region comprise a peptide sequence.

5. The isolated fusion protein of claim 4, wherein the peptide sequence comprises the amino acid sequence of SEQ ID NO:

43, SEQ ID NO;

44, SEQ ID NO;

45, or SEQ ID NO;

46.

6. The isolated fusion protein of claim 1, wherein the fusion protein further comprises a WAP domain-containing polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:

9-13.

7. The isolated fusion protein of claim 1, wherein the immunoglobulin Fc polypeptide is modified to enhance FcRn binding.

8. The isolated fusion protein of claim 1, wherein the immunoglobulin Fc polypeptide comprises at least one of the following mutations:

Met252Tyr, Ser254Thr, Thr256Glu, Met428Leu or Asn434Ser.

9. The isolated fusion protein of claim 1, wherein the immunoglobulin Fc polypeptide comprises at least one mutation at a position selected from the group consisting of:

Met252, Ser254, Thr256, Met428, and Asn434.

17. A method of treating or alleviating a symptom of a disease or disorder associated with aberrant serine protease expression or activity in a subject in need thereof, the method comprising administering a fusion protein according to claim 1 or claim 10.

18. The method of claim 17, wherein the subject is a human.

19. A method of treating or alleviating inflammation or a symptom of an inflammatory disease or disorder while reducing the risk of infection, in a subject in need thereof, the method comprising administering a fusion protein according to claim 1 or claim 10.

20. The method of claim 19, wherein the inflammatory disease or disorder is selected from the following:

emphysema, chronic obstructive pulmonary disease (COPD), acute respiratory distress syndrome (ARDS), allergic asthma, cystic fibrosis, cancers of the lung, ischemia-reperfusion injury, ischemia/reperfusion injury following cardiac transplantation, myocardial infarction, rheumatoid arthritis, septic arthritis, psoriatic arthritis, ankylosing spondylitis, Crohn'"'"'s disease, psoriasis, type I and/or type II diabetes, pneumonia, sepsis, graft versus host disease (GVHD), wound healing, Systemic lupus erythematosus, and Multiple sclerosis.

21. A method of reducing the risk of infection in a subject in need thereof, the method comprising administering a fusion protein according to claim 1 or claim 10.

22. The method of claim 21, wherein the infection is selected from bacterial infections, fungal infections, or viral infections.

10. An isolated fusion protein comprising at least one human alpha-1 antitrypsin (AAT) polypeptide comprising a human alpha-1 antitrypsin (AAT) polypeptide comprising the amino acid sequence of SEQ ID NO:

32 or SEQ ID NO;

33 operably linked to

an immunoglobulin Fc polypeptide comprising the amino acid sequence of SEQ ID NO;

6.

11. The isolated fusion protein of claim 10, wherein the AAT polypeptide and the immunoglobulin Fc polypeptide are operably linked via a hinge region, a linker region, or both a hinge region and linker region.

12. The isolated fusion protein of claim 10, wherein the fusion protein further comprises a SLPI polypeptide comprising the amino acid sequence of SEQ ID NO:

9 or SEQ ID NO;

10.

13. The isolated fusion protein of claim 11, wherein the hinge region, the linker region or both the hinge region and the linker region comprise a peptide sequence.

14. The isolated fusion protein of claim 10, wherein the immunoglobulin Fc polypeptide is modified to enhance FcRn binding.

15. The isolated fusion protein of claim 10, wherein the immunoglobulin Fc polypeptide comprises at least one mutation at a position selected from the group consisting of:

Met252, Ser254, Thr256, Met428, and Asn434.

16. The isolated fusion protein of claim 10, wherein the immunoglobulin Fc polypeptide comprises at least one of the following mutations:

Met252Tyr, Ser254Thr, Thr256Glu, Met428Leu or Asn434Ser.