Modified vaccinia ankara expressing p53 in cancer immunotherapy

• US 20040208850A1
• Filed: 12/23/2003
• Published: 10/21/2004
• Est. Priority Date: 12/23/2002
• Status: Active Grant

• Current Assignee: City of Hope
• Original Assignee: City of Hope
• Inventors: Ellenhorn, Joshua D.I., Diamond, Don J.
• Granted Patent: US 7,256,037 B2
• Time in Patent Office: Days
• US Class Current: 424/93.2
• CPC Class Codes: A61K 2039/5156 expressing foreign proteins
• CPC Class Codes: A61K 2039/55516 Proteins; Peptides
• CPC Class Codes: A61K 2039/55561 CpG containing adjuvants; Oligonucleotide containing adjuvants
• CPC Class Codes: A61K 2039/57 characterised by the type of response, e.g. Th1, Th2
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/4746 p53
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2710/24143 viral genome or elements thereof as genetic vector
• CPC Class Codes: Y10S 435/81 Packaged device or kit

Abstract

Mutations to the tumor suppressor protein p53 have been observed in 40-60% of all human cancers. These mutations are often associated with high nuclear and cytoplasmic concentrations of p53. Since many tumors exhibit highly elevated p53 levels, the protein is an attractive target for cancer immunotherapy. Unfortunately, p53 is an autoantigen that is likely to be tolerated as a self-protein by the immune system. The present invention is based on the discovery that this self-tolerance can be overcome by administration of recombinant modified vaccinia Ankara (MVA) containing a nucleic acid that encodes p53 (rMVAp53). The invention discloses a method of generating a p53-specific CTL-response to tumor cells expressing mutated p53 by administering a composition comprising rMVAp53. Administration of rMVAp53 decreases tumor development, tumor growth, and mortality in a variety of malignant cell types. These effects are enhanced by administration of CTLA-4 blocker and/or CpG oligodeoxynucleotide immunomodulators.

Claims

1. A composition comprising a recombinant MVA virus (rMVA) containing a nucleic acid sequence encoding p53.

2. The composition of claim 1, wherein said p53 is wild type murine p53.

3. The composition of claim 2, wherein said nucleic acid sequence comprises the nucleotide sequence of SEQ ID NO:

1.

4. The composition of claim 1, wherein said p53 is wild type human p53.

5. The composition of claim 4, wherein said nucleic acid sequence comprises the nucleotide sequence of SEQ ID NO:

2.

6. The composition of claim 1, further comprising an immunomodulator comprising a CTLA-4 blocking agent, a CpG oligodeoxynucleotide, or both a CTLA-4 blocking agent and a CpG oligodeoxynucleotide.

7. The composition of claim 6, wherein said CTLA-4 blocking agent is an antibody.

8. The composition of claim 7, wherein said antibody is a monoclonal antibody.

9. A method of treating a subject having a p53-expressing malignancy, comprising introducing into said subject a composition comprising recombinant MVA virus containing a nucleic acid sequence encoding p53.

10. The method of claim 9, wherein said subject is human.

11. The method of claim 9, wherein introduction of said composition elicits an immune response effective against said p53-expressing malignancy.

12. The method of claim 9, wherein said p53 is wild type human p53.

13. The method of claim 12, wherein said nucleic acid sequence comprises the nucleotide sequence of SEQ ID NO:

2.

14. The method of claim 9, wherein introduction of said composition is repeated one or more times.

15. The method of claim 9, further comprising introducing into said subject an immunomodulator comprising a CTLA-4 blocking agent, a CpG oligodeoxynucleotide, or both a CTLA-4 blocking agent and a CpG oligodeoxynucleotide.

16. The method of claim 15, wherein said CTLA-4 blocking agent is an antibody.

17. The method of claim 16, wherein said antibody is a monoclonal antibody.

18. The method of claim 15, wherein introduction of said immunomodulator occurs prior to, simultaneous with, or after introduction of the composition comprising recombinant MVA virus.

19. The method of claim 15, wherein introduction of said immunomodulator is repeated one or more times.

20. The method of claims 9 or 15, wherein the method of said introduction is selected from the group consisting of subcutaneous, percutaneous, intradermal, intraperitoneal, intramuscular, intratumoral and intravenous injection.

21. A kit for treating a subject having a p53-expressing malignancy comprising a composition comprising recombinant MVA virus containing a nucleic acid sequence encoding p53 and instructions for administration of said composition, wherein said administration elicits an immune response effective against said p53-expressing malignancy.

22. The kit of claim 21, wherein said p53 is human p53.

23. The kit of claim 22, wherein said nucleic acid sequence comprises the nucleotide sequence of SEQ ID NO:

2.

24. The kit of claim 21, further comprising an immunomodulator comprising a CTLA4 blocking agent, a CpG oligodeoxynucleotide, or both a CTLA-4 blocking agent and a CpG oligodeoxynucleotide.

25. The kit of claim 24, wherein said CTLA-4 blocking agent is an antibody.

26. The kit of claim 25, wherein said antibody is a monoclonal antibody.

27. The kit of claim 21, wherein said subject is human.

28. A vector comprising an MVA recombination plasmid containing a nucleic acid insert encoding p53.

29. The vector of claim 28, wherein said MVA recombination plasmid is pLW22.

30. The vector of claim 28, wherein said nucleic acid insert encodes human wild type p53.

31. The vector of claim 30, wherein said vector comprises the sequence of SEQ ID NO:

5.

32. A method of generating a p53 specific cytotoxic T lymphocyte (CTL) response against cells overexpressing mutant p53 comprising administering a composition comprising recombinant MVA virus containing a nucleic acid sequence encoding p53.

33. The method of claim 32, wherein said nucleic acid sequence encoding p53 encodes murine wild type p53.

34. The method of claim 33, wherein said nucleic acid sequence comprises the nucleotide sequence of SEQ ID NO:

1.

35. The method of claim 32, wherein said nucleic acid sequence encoding p53 encodes human wild type p53.

36. The method of claim 35, wherein said nucleic acid sequence comprises the nucleotide sequence of SEQ ID NO:

2.

37. The method of claim 32, further comprising administering an immunomodulator comprising a CTLA-4 blocking agent, a CpG oligodeoxynucleotide, or both a CTLA-4 blocking agent and a CpG oligodeoxynucleotide.

38. The method of claim 37, wherein said CTLA-4 blocking agent is an antibody.

39. The method of claim 38, wherein said antibody is a monoclonal antibody.

40. The method of claim 37, wherein administration of said immunomodulator occurs prior to, simultaneous with, or after introduction of the composition comprising recombinant MVA virus.

41. The method of claim 37, wherein introduction of said immunomodulator is repeated one or more times.