Compositions, methods and kits for treatment of cancer and autoimmune diseases

• US 9,447,160 B2
• Filed: 01/18/2013
• Issued: 09/20/2016
• Est. Priority Date: 01/19/2012
• Status: Active Grant

• Current Assignee: University of Miami
• Original Assignee: University of Miami
• Inventors: Bhatt, Shruti, Lossos, Izidore, Rosenblatt, Joseph, Shin, Seung-Uon
• Primary Examiner(s): LIEB, JEANETTE M
• Application Number:
  US14/373,137
• Publication Number:
  US 20150044134A1
• Time in Patent Office: 1,341 Days
• Field of Search: None
• US Class Current: 1/1
• 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 38/2013 IL-2
• CPC Class Codes: A61K 39/39558 against tumor tissues, cells, antigens
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 51/00 Preparations containing radioactive substances for use in therapy or testing in vivo
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07K 14/54 Interleukins [IL]
• CPC Class Codes: C07K 14/55 IL-2
• CPC Class Codes: C07K 16/2887 against CD20
• CPC Class Codes: C07K 2317/54 F(ab')2
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/732 Antibody-dependent cellular cytotoxicity [ADCC]
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/33 fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies

Abstract

The present invention is directed to fusion proteins having an IL-21 cytokine portion and an anti-CD20 antibody portion, and methods of using such fusion proteins. The invention also provides pharmaceutical compositions and kits utilizing the IL-21-anti-CD20 fusion proteins. In particular, the methods, kits and compositions of the invention are useful in the treatment of cancer and autoimmune diseases.

Claims

1. A recombinant fusion protein comprising an amino acid sequence of IL-21 or cytotoxic fragment thereof fused to an N-terminus of an amino acid sequence of an anti-CD20 antibody or antigen binding fragment thereof via a linker peptide of SEQ ID NO:

1.

2. The fusion protein of claim 1 comprising an amino acid sequence as set forth in SEQ ID NO:

14.

3. The fusion protein of claim 1 wherein the anti-CD20 antibody or antigen binding fragment thereof specifically binds one or more epitopes comprising the amino acid sequence set forth in SEQ ID NO:

2 or SEQ ID NO;

3, or combinations thereof.

4. The fusion protein of claim 1 wherein the anti-CD20 antibody or antigen binding fragment thereof specifically binds a polypeptide having an amino acid sequence as set forth in SEQ ID NO:

4, or portions thereof.

5. The fusion protein of claim 1, wherein the anti-CD20 antibody is a monoclonal antibody.

6. The fusion protein of claim 1, wherein the antigen-binding fragment of the anti-CD20 antibody is selected from the group consisting of Fab, Fab′

, F(ab′

)2, Fv fragments, diabodies, single chain antibody molecules, and multispecific antibodies formed from antibody fragments.

7. The fusion protein of claim 1 further comprising a label.

8. The fusion protein of claim 7, wherein the label is a radiolabel.

9. An isolated polynucleotide comprising a nucleic acid sequence encoding the fusion protein of claim 1.

10. A cloning vector comprising the nucleotide sequence of claim 9.

11. An expression vector comprising the nucleotide sequence of claim 9.

12. An expression vector encoding the fusion protein of claim 1.

13. A cultured cell expressing the fusion protein of claim 1.

14. A pharmaceutical composition comprising the fusion protein of claim 1 and at least one pharmaceutically acceptable carrier.

15. The pharmaceutical composition of claim 14, further comprising one or more pharmaceutically acceptable excipient, additive, or adjuvant.

16. The pharmaceutical composition of claim 14 further comprising at least one of a chemotherapeutic agent, a radioactive isotope, a therapeutic agent, or combinations thereof.

17. A kit comprising the pharmaceutical composition of claim 14.

18. A method of treating CD20 expressing leukemia or lymphoma in a subject comprising administering a therapeutically effective amount of the fusion protein of claim 1 to the subject.

19. A method of treating an autoimmune disease in a subject comprising administering a therapeutically effective amount of the fusion protein of claim 1 to the subject, wherein the autoimmune disease is affected by modulation of natural killer (NK) cells, cytotoxic T cells or B-lymphocytes.

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

21. The method of claim 18 further comprising administering to the subject a therapeutically effective amount of at least one agent selected from the group consisting of a chemotherapeutic agent, a radioactive isotope, a therapeutic agent, or combinations thereof.

22. The method of claim 18 wherein the fusion protein is administered with a pharmaceutically acceptable carrier.

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

24. The kit of claim 23 further comprising at least one agent capable of facilitating detection of the fusion protein.

25. A method of targeted delivery to one or more CD20 expressing tumors in a subject comprising administering a therapeutically effective amount of the fusion protein of claim 1 to the subject.

26. The method of claim 25 further comprising activating one or more immune effector cells adjacent the one or more tumors by the recombinant fusion protein.

27. A method of inhibiting the growth of a CD20 expressing leukemia or lymphatic tumor cell comprising contacting the tumor cell with the fusion protein of claim 1.

28. A method of targeting CD20 and IL-21 receptors on a cell with a single protein, comprising contacting the cell with the fusion protein of claim 1.

29. A method of enhancing antibody dependent cellular cytotoxicity activity against CD20 expressing leukemia or lymphatic cells expressing CD20 in a subject comprising administering the fusion protein of claim 1 to the subject.

30. The method of claim 19 wherein the subject is a human.

31. The method of claim 19 further comprising administering to the subject a therapeutically effective amount of at least one agent selected from the group consisting of a chemotherapeutic agent, a radioactive isotope, a therapeutic agent, or combinations thereof.

32. The method of claim 19 wherein the fusion protein is administered with a pharmaceutically acceptable carrier.