Protein inhibitors to complement and VEGF pathways and methods of use thereof

• US 9,988,611 B2
• Filed: 11/30/2012
• Issued: 06/05/2018
• Est. Priority Date: 12/01/2011
• Status: Active Grant

• Current Assignee: Innovent Biologics (Suzhou) Co. Ltd., AP Biosciences, Inc. (OBI Pharma Inc.)
• Original Assignee: Innovent Biologics (Suzhou) Co. Ltd., AP Biosciences, Inc. (OBI Pharma Inc.)
• Inventors: Her, Jeng-Horng, Chen, Huang-Tsu
• Primary Examiner(s): PAK, MICHAEL D
• Application Number:
  US14/362,109
• Publication Number:
  US 20150079084A1
• Time in Patent Office: 2,013 Days
• Field of Search: None
• 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 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• 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 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• 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 14/472 Complement proteins, e.g. anaphylatoxin, C3a, C5a
• CPC Class Codes: C07K 14/70596 Molecules with a "CD"-designation not provided for elsewhere
• CPC Class Codes: C07K 14/71 for growth factors; for growth regulators
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/02 containing a signal sequence
• 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 2319/70 containing domain for protein-protein interaction
• CPC Class Codes: C12N 9/1205 Phosphotransferases with an alcohol group as acceptor (2.7.1), e.g. protein kinases
• CPC Class Codes: C12Y 207/01112 : Protein-tyrosine kinase (2.7.1.112) C12Y207/10001, C12Y207/10002 take precedence

Abstract

The invention provides bispecific fusion proteins that inhibit activation of complement pathway and vascular endothelial growth factor (VEGF) pathway and methods for using these fusion proteins.

Claims

1. A fusion protein comprising a complement inhibiting domain (CID), a VEGF inhibiting domain (VID), and a half-life prolonging domain, wherein the fusion protein inhibits complement activation and VEGF activity, wherein the CID comprises an amino acid sequence selected from the group consisting of SEQ ID NO:

1-4, or an amino acid sequence having at least 98% identity to an amino acid sequence selected from the group consisting of SEQ ID NO;

1-4, and wherein the VID comprises the amino acid sequence of SEQ ID NO;

11 or 38, or an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO;

11 or 38.

2. The fusion protein of claim 1, wherein the CID comprises an amino acid sequence selected from the group consisting of SEQ ID NO:

1-4.

3. The fusion protein of claim 1, wherein the VID comprises the amino acid sequence of SEQ ID NO:

11 or 38.

4. The fusion protein of claim 1, wherein the half-life prolonging domain comprises an immunoglobulin Fc region.

5. The fusion protein of claim 4, wherein the Fc region is a human Fc of IgG1, IgG2, IgG3, or IgG4.

6. The fusion protein of claim 4, wherein the Fc region comprises the amino acid sequence of SEQ ID NO:

7 or 39, or an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO;

7 or 39.

7. The fusion protein of claim 1, where the fusion protein further comprises a peptide linker between domains.

8. The fusion protein of claim 7, wherein the peptide linker comprises the amino acid sequence of SEQ ID NO:

8.

9. The fusion protein of claim 4, wherein the fusion protein comprises said VID, CID, and Fc from N-terminal to C-terminal in an order selected from the group consisting of (1) VID, Fc, CID;

(2) CID, Fc, VID;

(3) CID, VID, Fc;

(4) VID, CID, Fc;

(5) Fc, VID, CID; and

(6) Fc, CID, VID.

19. A fusion protein produced by culturing a host cell comprising a nucleic acid encoding the fusion protein of claim 1 under a condition that produces the fusion protein, and recovering the fusion protein produced by the host cell.

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

22. The composition of claim 21, wherein the fusion protein is a dimeric form.

23. A method of producing a fusion protein comprising culturing a host cell comprising a nucleic acid encoding the fusion protein of claim 1 under a condition that produces the fusion protein, and recovering the fusion protein produced by the host cell.

24. The method of claim 23, wherein the fusion protein is recovered from the cell culture medium and purified.

25. The method of claim 23, wherein the host cell is a mammalian cell or a yeast cell.

26. The method of claim 23, wherein the fusion protein recovered is a dimer.

27. A kit comprising the fusion protein of claim 1.

28. The kit of claim 27, further comprising a package insert comprising instructions for use the fusion protein for treating an inflammatory disease, an autoimmune disease, an ocular disease or cancer in a subject.

10. A fusion protein comprising, from the N-terminal to C-terminal, a VEGF inhibiting domain (VID), an immunoglobulin Fc region, and a complement inhibiting domain (CID), wherein the fusion protein inhibits complement activation and VEGF activity, wherein the CID comprises an amino acid sequence selected from the group consisting of SEQ ID NO:

1-4, or an amino acid sequence having at least 98% identity to an amino acid sequence selected from the group consisting of SEQ ID NO;

1-4, and wherein the VID comprises the amino acid sequence of SEQ ID NO;

11 or 38, or an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO;

11 or 38.

11. The fusion protein of claim 10, wherein the CID comprises an amino acid sequence selected from the group consisting of SEQ ID NO:

1-4.

12. The fusion protein of claim 10, wherein the VID comprises the amino acid sequence of SEQ ID NO:

11 or 38.

13. The fusion protein of claim 10, wherein the Fc region is a human Fc of IgG1, IgG2, IgG3 or IgG4.

14. The fusion protein of claim 13, wherein the Fc region comprises the sequence of SEQ ID NO:

7 or 39, or an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO;

7 or 39.

15. The fusion protein of claim 10, where the fusion protein further comprises a peptide linker between domains.

16. The fusion protein of claim 15, wherein the peptide linker is between the Fc region and the CID.

17. The fusion protein of claim 15, wherein the peptide linker comprises the amino acid sequence of SEQ ID NO:

8.

18. The fusion protein of claim 10, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO:

12 or 40, or an amino acid sequence having at least 98% identity to the amino acid sequence of SEQ ID NO;

12 or 40.

20. A dimeric fusion protein comprising two fusion proteins, wherein each fusion protein comprises the fusion protein of claim 10.

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

30. The composition of claim 29, wherein the fusion protein is a dimeric form.

31. The fusion protein of claim 10, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO:

12 or 40.

34. A composition comprising the fusion protein of claim 31 and a pharmaceutically acceptable carrier.

35. The composition of claim 34, wherein the fusion protein is a dimeric form.

40. A kit comprising the fusion protein of claim 31.

41. The kit of claim 40, further comprising a package insert comprising instructions for use the fusion protein for treating an inflammatory disease, an autoimmune disease, an ocular disease or cancer in a subject.

32. A fusion protein produced by culturing a host cell comprising a nucleic acid encoding a fusion protein comprising the amino acid sequence of SEQ ID NO:

12 or 40 under a condition that produces the fusion protein, and recovering the fusion protein produced by the host cell.

33. A dimeric fusion protein comprising two fusion proteins, wherein each fusion protein comprises the amino acid sequence of SEQ ID NO:

12 or 40.

36. A method of producing a fusion protein comprising culturing a host cell comprising a nucleic acid encoding the amino acid sequence of SEQ ID NO:

12 or 40 under a condition that produces the fusion protein, and recovering the fusion protein produced by the host cell.

37. The method of claim 36, wherein the fusion protein is recovered from the cell culture medium and purified.

38. The method of claim 36, wherein the host cell is a mammalian cell or a yeast cell.

39. The method of claim 36, wherein the fusion protein recovered is a dimer.