TARGETED INTERFERONS DEMONSTRATE POTENT APOPTOTIC AND ANTI-TUMOR ACTIVITIES

• US 20110104112A1
• Filed: 01/05/2011
• Published: 05/05/2011
• 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 8,563,692 B2
• Time in Patent Office: Days
• US Class Current: 424/85.4
• 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 composition comprising an interferon attached to a targeting moiety that binds to an antigen expressed in or associated with a cell, wherein the targeting moiety is attached to the interferon by a proteolysis resistant peptide linker, wherein the linker comprises at least one amino acid residue.

2. The composition of claim 1, wherein the linker comprises fewer than fifteen amino acids.

3. The composition of claim 1, wherein the linker comprises ten or fewer amino acids.

4. The composition of claim 1, wherein the linker comprises five or fewer amino acids.

5. The composition of claim 1, wherein the linker is not (Gly₄Ser)₃ (SEQ ID NO:

5).

6. The composition of claim 1, wherein the linker comprises 27 or fewer amino acids.

7. The composition of claim 1, wherein the linker is selected from the group consisting of:

8. The composition of claim 1, wherein the linker is selected from the group consisting of:

9. The composition of claim 1, wherein the targeting moiety is an antibody.

10. The composition of claim 1, wherein the targeting moiety is an antibody that binds a tumor associated antigen.

11. An isolated nucleic acid encoding the composition of claim 1.

12. A plasmid comprising the isolated nucleic acid of claim 11.

13. A cell comprising the isolated nucleic acid of claim 11.

14. A kit comprising the composition of claim 1.

15. The composition of claim 1, wherein the antigen is a cancer marker.

16. The composition of claim 1, wherein the targeting moiety is an anti-CD20 antibody, the interferon is IFNα

or IFNβ

, and the linker comprises fewer than fifteen amino acids.

17. The composition of claim 1, wherein the targeting moiety is an anti-CD20 antibody, the interferon is IFNα

or IFNβ

, and the linker does not comprise (Gly₄Ser)₃ (SEQ ID NO;

5).

18. The composition of claim 1, wherein the targeting moiety is an anti-CD20 antibody, the interferon is IFNα

or IFNβ

, and the linker is GGGGS (SEQ ID NO;

6).

19. The composition of claim 1, wherein the targeting moiety is an anti-CD20 antibody, the interferon is IFNα

or IFNβ

, and the linker is AEAAAKEAAAKA (SEQ ID NO;

7).

20. The composition of claim 16, wherein the anti-CD20 antibody is RITUXIMAB®

.

21. The composition of claim 1, wherein the targeting moiety is an anti-EGFR family antibody, the interferon is IFNα

or IFNβ

, and the linker is GGGGS (SEQ ID NO;

6).

22. The composition of claim 21, wherein the anti-EGFR family antibody is an anti-HER2/neu antibody.

23. The composition of claim 1, wherein the targeting moiety is an anti-EGFR family antibody, the interferon is IFNα

or IFNβ

, and the linker is

24. The composition of claim 23, wherein the anti-EGFR family antibody is an anti-HER2/neu antibody.

25. A method of treating cancer comprising administering to a subject the composition of claim 1.

26. A method for killing or inhibiting the growth or proliferation of a cancer cell, comprising:

contacting a cancer cell with an interferon attached to a targeting moiety that binds to a cancer marker expressed in or associated with the cancer cell, wherein the targeting moiety is attached to the interferon by a proteolysis resistant peptide linker, wherein the linker comprises at least one amino acid residue; and

killing or inhibiting the growth or proliferation of the cancer cell.

27. The method of claim 26, wherein the interferon is IFNα

or IFNβ

.

28. A method for increasing therapeutic efficacy of an interferon for treatment of cancer, comprising:

administering to a subject an interferon attached to a targeting moiety that binds to a tumor associated antigen expressed on or associated with a cancer cell in the subject, wherein the targeting moiety is attached to the interferon by a proteolysis resistant peptide linker, wherein the linker comprises at least one amino acid residue;

wherein the interferon attached to the targeting moiety is more effective in treating the cancer than the interferon not attached to the targeting moiety.

29. The method of claim 28, wherein the interferon is IFNα

or IFNβ

.

30. A method for reducing toxicity associated with administering an interferon to a subject, comprising:

administering to a subject an interferon attached to a targeting moiety that binds to a tumor associated antigen expressed in or associated with a cancer cell in the subject, wherein the targeting moiety is attached to the interferon by a proteolysis resistant linker, wherein the linker comprises at least one amino acid residue;

wherein administering the interferon attached to the targeting moiety causes less toxicity in the subject than administering the interferon not attached to the targeting moiety.

31. The method of claim 30, wherein the interferon is IFNα

or IFNβ

.

32. A method for increasing the in vivo half-life of an interferon upon administration to a subject, comprising:

preparing or obtaining a composition comprising an interferon attached to a targeting moiety that binds to an antigen expressed in or associated with a cell, wherein the targeting moiety is attached to the interferon by a proteolysis resistant peptide linker, wherein the linker comprises at least one amino acid residue;

identifying a subject that has been previously administered the interferon not attached to the targeting moiety by the linker; and

administering to the subject the interferon attached to the targeting moiety by the linker.

33. The method of claim 32, wherein the interferon is IFNα

or IFNβ

.

34. A method for treating cancer in a subject resistant to treatment with an antibody, an interferon or both, comprising:

identifying a subject with cancer, wherein the subject initially responds to treatment with an antibody, an interferon or both, and wherein the subject subsequently becomes less sensitive to the treatment;

administering to the subject a composition comprising the interferon attached to the antibody, wherein the antibody is attached to the interferon by a proteolysis resistant peptide linker, wherein the linker comprises at least one amino acid residue

35. The method of claim 34, wherein the antibody is an anti-CD20 antibody.

36. The method of claim 35, wherein the anti-CD20 antibody is RITUXIMAB®

.

37. A method for increasing therapeutic efficacy of an interferon for the treatment of cancer, comprising

preparing or obtaining a composition comprising an interferon attached to a targeting moiety that binds to a tumor associated antigen, wherein the targeting moiety is attached to the interferon by a proteolysis resistant peptide linker, wherein the linker comprises at least one amino acid residue;

and

administering to a subject the interferon attached to the targeting moiety by the linker,

wherein the interferon attached to the targeting moiety is more effective in treating cancer in the subject than the interferon not attached to the targeting moiety.