Recombinant herpes simplex virus useful for treating neoplastic disease

• US 7,744,899 B2
• Filed: 10/11/2006
• Issued: 06/29/2010
• Est. Priority Date: 12/31/1998
• Status: Expired due to Fees

• Current Assignee: UAB Research Foundation (University of Alabama System), University of Chicago
• Original Assignee: UAB Research Foundation (University of Alabama System), University of Chicago
• Inventors: Roizman, Bernard, Whitley, Richard J.
• Primary Examiner(s): WEHBE, ANNE MARIE SABRINA
• Application Number:
  US11/546,233
• Publication Number:
  US 20070031383A1
• Time in Patent Office: 1,357 Days
• Field of Search: 424/199.1, 514/44, 435/320.1
• US Class Current: 424/199.1
• CPC Class Codes: A61K 35/763 Herpes virus
• 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 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/5406 IL-4
• CPC Class Codes: C07K 14/5428 IL-10
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2710/16632 Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
• CPC Class Codes: C12N 2710/16643 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 7/00 Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00

Abstract

Recombinant herpes simplex viruses comprising DNA encoding cytokines and methods for treating neoplastic diseases using the inventive recombinant viruses are disclosed.

Claims

1. A recombinant herpes simplex virus that preferentially kills tumor cells and not healthy cells, that expresses only one γ

₁34.5 gene copy and that comprises an expressible tumor-growth-inhibiting cytokine-encoding DNA.

2. The recombinant herpes simplex virus of claim 1 wherein said virus lacks all or part of a γ

₁34.5 gene copy.

3. The recombinant herpes simplex virus of claim 2 wherein said virus comprises a γ

₁34.5 gene having a deletion of a portion of a coding sequence of said γ

₁34.5 gene, said deletion comprising a Bst EII-Stu I fragment of said γ

₁34.5 gene.

4. The recombinant herpes simplex virus of claim 1 wherein said virus comprises a γ

₁34.5 gene having a stop codon at a Bst EII site in said γ

₁34.5 gene.

5. The recombinant herpes simplex virus of claim 1 wherein said expressible tumor-growth-inhibiting cytokine-encoding DNA is under the promoter-regulatory control of a herpes simplex virus gene promoter.

6. The recombinant herpes simplex virus of claim 5 wherein said herpes simplex virus gene promoter is an EGR-1 promoter.

7. The recombinant herpes simplex virus of claim 1 wherein said tumor-growth-inhibiting cytokine-encoding DNA is under the promoter-regulatory control of a synthetic herpes simplex virus-derived promoter.

8. The recombinant herpes simplex virus of claim 7 wherein said synthetic herpes simplex virus-derived promoter comprises a herpes simplex virus α

gene promoter fragment operatively linked 5′

to a herpes simplex virus γ

gene promoter fragment.

9. The recombinant herpes simplex virus of claim 8 wherein said α

gene promoter fragment comprises promoter sequences upstream of the transcription initiation site of the α

4 gene and said γ

gene promoter fragment comprises a transcription initiation site and the 5′

transcribed non-coding sequence of the γ

₁U_L9 gene.

10. The recombinant herpes simplex virus of claim 1 wherein an unexpressed γ

₁34.5 gene copy is replaced by said expressible tumor-growth-inhibiting cytokine-encoding DNA.

11. The recombinant herpes simplex virus of claim 1 wherein said virus comprises two or more copies of said tumor-growth-inhibiting cytokine-encoding DNA.

12. The recombinant herpes simplex virus of claim 10 where said herpes simplex virus is a HSV-1.

13. The recombinant virus of claim 1 wherein said tumor-growth-inhibiting cytokine-encoding DNA further comprises a polyadenylation signal.

14. The recombinant virus of claim 13 wherein said polyadenylation signal is a hepatitis B virus-derived polyadenylation signal.

27. The recombinant herpes simplex virus of claim 3, wherein the expressible tumor growth inhibiting cytokine-encoding DNA is selected from DNA encoding IL-1, IL-2, IL-A, IL-6, IL-7, IFN-γ

, GM-CSF or TNF-α

.

15. A method for treating neoplastic disease, the method comprising administering to a target tumor, a recombinant herpes simplex virus capable of expressing only one γ

₁34.5 gene copy and comprising an expressible tumor-growth-inhibiting cytokine-encoding DNA, wherein the expressed cytokine augments tumor cell killing.

16. The method of claim 15 wherein said recombinant herpes simplex virus lacks all or part of an unexpressed γ

₁34.5 gene copy.

17. The method of claim 15 wherein said recombinant herpes simplex virus comprises a γ

₁34.5 gene having a stop codon at a Bst EII site in said γ

₁34.5 gene.

18. The method of claim 15 wherein said recombinant herpes simplex virus comprises a γ

₁34.5 gene lacking a portion of the coding sequence corresponding to a Bst EII/Stu I restriction fragment of said γ

₁34.5 gene.

19. The method of claim 15 wherein said expressible tumor-growth-inhibiting cytokine-encoding DNA is under the promoter-regulatory control of a herpes simplex virus gene promoter.

20. The method of claim 19 wherein said herpes simplex virus promoter is an EGR-1 promoter.

21. The method of claim 15 wherein said tumor-growth-inhibiting cytokine-encoding DNA is under the promoter regulatory control of a synthetic herpes simplex virus-derived promoter.

22. The method of claim 21 wherein said synthetic herpes simplex virus-derived promoter comprises a herpes simplex virus α

gene fragment operatively linked 5′

to a herpes simplex virus γ

gene promoter fragment.

23. The method of claim 22 wherein said α

gene promoter fragment comprises a promoter sequence upstream of the transcription initiation site of said α

gene promoter fragment comprising the transcription initiation site and the 5′

transcribed non-coding sequence of the U_L19 gene.

24. The method of claim 15 wherein said γ

₁34.5 gene is replaced by said expressible tumor-growth-inhibiting cytokine-encoding DNA.

26. The method of claim 15, wherein the target tumor is a tumor of the central nervous system.

28. The method of claim 15, wherein the expressible tumor growth inhibiting cytokine-encoding DNA is selected from DNA encoding IL-1, IL-2, IL-4, IL-6, IL-7, IFN-γ

, GM-CSF or TNF-α

.

25. A pharmaceutical composition comprising in a pharmaceutically acceptable carrier, diluent, or adjuvant, a recombinant herpes simplex virus expressing only one γ

₁34.5 gene copy, said virus comprising an expressible tumor-growth-inhibiting cytokine-encoding DNA, wherein the expressed cytokine augments tumor cell killing.