TARGETED INTERFERONS DEMONSTRATE POTENT APOPTOTIC AND ANTI-TUMOR ACTIVITIES

• US 20100172868A1
• Filed: 12/30/2009
• Published: 07/08/2010
• Est. Priority Date: 09/21/2007
• Status: Active Grant

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: HUANG, TZU-HSUAN, MORRISON, SHERIE L., XUAN, CAIYUN
• Granted Patent: US 8,258,263 B2
• Time in Patent Office: Days
• US Class Current: 424/85.400
• CPC Class Codes: A61K 2039/505 comprising antibodies
• 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: A61K 38/212 IFN-alpha
• CPC Class Codes: A61K 38/215 IFN-beta
• CPC Class Codes: A61K 47/6813 the drug being a peptidic cytokine, e.g. an interleukin or interferon
• CPC Class Codes: A61K 47/6851 the antibody targeting a determinant of a tumour cell
• CPC Class Codes: A61K 47/6855 the tumour determinant being from breast cancer cell
• CPC Class Codes: A61K 47/6875 the antibody being a hybrid immunoglobulin
• 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 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: C07K 14/555 Interferons [IFN]
• CPC Class Codes: C07K 14/56 IFN-alpha
• CPC Class Codes: C07K 14/565 IFN-beta
• CPC Class Codes: C07K 16/2863 against receptors for growth factors, growth regulators
• CPC Class Codes: C07K 16/2887 against CD20
• CPC Class Codes: C07K 16/32 against translation products of oncogenes
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2319/33 : fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies

Abstract

Novel chimeric moieties that show significant efficacy against cancers are provided. In certain embodiments the chimeric moieties comprise a targeting moiety attached to an interferon. In certain embodiments, the chimeric moieties comprise fusion proteins where an antibody that specifically binds to a cancer marker is fused to interferon alpha (IFN-α) or interferon beta (IFN-β).

Claims

1. A chimeric construct comprising an interferon attached to a targeting moiety that binds to tumor associated antigen, wherein said targeting moiety is attached directly to said interferon or by a linker that is a proteolysis resistant linker, wherein said construct when contacted to a tumor cell results in the killing or inhibition of growth or proliferation of said tumor cell.

2. The construct of claim 1, wherein said targeting moiety is attached to said interferon by a linker that is not a (Gly4Ser)₃ (SEQ ID NO:

5) linker.

3. The construct of claim 1, wherein said linker is fewer than 15 amino acids in length.

4. The construct of claim 1, wherein said targeting moiety is attached to said interferon by a linker that is about the length of Gly₄Ser (SEQ ID NO:

6) or shorter.

5. The construct of claim 1, wherein said targeting moiety is attached to said interferon by a Gly₄Ser (SEQ ID NO:

6) linker.

6. The construct according to claim 1, wherein said interferon is a type 1 interferon.

7. The construct according to claim 1, wherein said interferon is a type 2 interferon.

8. The construct according to claim 1, wherein said interferon is an interferon alpha.

9. The construct according to claim 1, wherein said interferon is an interferon beta.

10. The construct according to claim 1, wherein the activity of said construct is at least 2-fold greater in vitro against a cell expressing the target marker than the same interferon when not attached to a targeting moiety.

11. The construct according to claim 1, wherein said targeting moiety is chemically coupled to said interferon.

12. The construct according to claim 1, wherein said targeting moiety is joined to said interferon with a peptide linker.

13. The construct of claim 12, wherein said construct is a recombinantly expressed fusion protein.

14. The construct according to claim 1, wherein said targeting moiety is an antibody that binds a tumor associated antigen.

15. The construct of claim 14, wherein said antibody specifically binds a marker selected from the group consisting of HER3, HER2/neu, MUC-1, G250, mesothelin, gp100, tyrosinase, and MAGE.

16. The construct of claim 14, wherein said targeting moiety is an antibody that binds CD20.

17. The construct of claim 16, wherein said targeting moiety is an antibody that comprises the complementarity determining regions (CDRs) of anti-CD20 (RITUXIMAB®

).

18. The construct of claim 16, wherein said targeting moiety is an antibody that comprises the variable regions of anti-CD20 (RITUXIMAB®

).

19. The construct of claim 15, wherein said targeting moiety is an antibody that binds HER2.

20. The construct of claim 19, wherein said antibody is a C6 antibody.

21. The construct of claim 19, wherein said antibody comprises the VH and VL CDRs of C6 MH3-B1.

22. The construct of claim 19, wherein said antibody comprises the VH and VL domains of C6 MH3-B1.

23. The construct of claim 14, wherein said antibody is an antibody selected from the group consisting of is a single chain Fv (scFv), a FAB, a (Fab′

)2, an (ScFv)2, and a full IgG.

24. The construct of claim 14, wherein said antibody is an antibody selected form the group consisting of Rituxan, IFS, B1, 1H4, CD19, B4, B43,FVS191, hLL2, LL2, RFB4, M195, HuM195, AT13/5, Herceptin, 4D5, HuCC49, HUCC39Δ

CH2 B72.3, 12C10, IG5, H23, BM-2, BM-7, 12H12, MAM-6, HMFG-1.

25. The construct of claim 14, wherein said antibody is an antibody that binds a member of the EGF receptor family.

26. The construct of claim 25, wherein said antibody is selected from the group consisting of C6.5, C6mL3-9, C6 MH3-B1, C6-B1D2, F5, HER3.A5, HER3.F4, HER3.H1, HER3.H3, HER3.E12, HER3.B12, EGFR.E12, EGFR.C10, EGFR.B11, EGFR.E8, HER4.B4, HER4.G4, HER4.F4, HER4.A8, HER4.B6, HER4.D4, HER4.D7, HER4.D11, HER4.D12, HER4.E3, HER4.E7, HER4.F8 and HER4.C7.

27. A pharmaceutical formulation comprising a construct according to claim 1 in a pharmaceutically acceptable excipient.

28. The pharmaceutical formulation according to claim 27, wherein said formulation is a unit dosage formulation.

29. The pharmaceutical formulation according to claim 27, wherein said formulation is a formulated for parenteral administration.

30. The pharmaceutical formulation according to claim 27, wherein said formulation is a formulated for administration via a route selected from the group consisting of oral administration, intravenous administration, intramuscular administration, direct tumor administration, inhalation, rectal administration, vaginal administration, transdermal administration, and subcutaneous depot administration.

31. A method of inhibiting growth and/or proliferation of a cancer cell, said method comprising contacting said cancer cell with a chimeric construct according to claim 1.

36. The method of claim 31, wherein said cancer cell is cell produced by a cancer selected from the group consisting of a B cell lymphoma, lung cancer, a bronchus cancer, a colorectal cancer, a prostate cancer, a breast cancer, a pancreas cancer, a stomach cancer, an ovarian cancer, a urinary bladder cancer, a brain or central nervous system cancer, a peripheral nervous system cancer, an esophageal cancer, a cervical cancer, a melanoma, a uterine or endometrial cancer, a cancer of the oral cavity or pharynx, a liver cancer, a kidney cancer, a biliary tract cancer, a small bowel or appendix cancer, a salivary gland cancer, a thyroid gland cancer, a adrenal gland cancer, an osteosarcoma, a chondrosarcoma, a liposarcoma, a testes cancer, and a malignant fibrous histiocytoma.

37. The method of claim 31, wherein said contacting comprises systemically administering said chimeric moiety to a mammal.

38. The method of claim 31, wherein said contacting comprises administering said chimeric moiety directly into a tumor site.

39. The method of claim 31, wherein said contacting comprises intravenous administration of said chimeric moiety.

32-35. -35. (canceled)

40-41. -41. (canceled)

42. A nucleic acid that encodes a fusion protein, said fusion protein comprising an interferon attached to a C6 single-chain antibody or to an anti-CD20 single chain antibody.

43-51. -51. (canceled)

52. A cell comprising a nucleic acid that expresses a fusion protein, wherein said nucleic acid encodes a fusion protein comprising an interferon attached to a C6 single-chain antibody or to an anti-CD20 single chain antibody.

53. (canceled)