Claims
- 1. An adenoviral vector comprising (a) an adenoviral genome deficient in the E4 region of the adenoviral genome, and optionally deficient in the E1 region, the E2A region, and/or the E3 region of the adenoviral genome, (b) a nucleic acid sequence coding for TNF, and (c) a radiation inducible promoter operably linked to the nucleic acid sequence coding for TNF.
- 2. The adenoviral vector of claim 1, wherein the adenoviral genome is deficient in the E1 region of the adenoviral genome.
- 3. The adenoviral vector of claim 2, wherein the nucleic acid sequence coding for TNF is located in the E1 region of the adenoviral genome.
- 4. The adenoviral vector of claim 3, wherein the TNF is TNF-α.
- 5. The adenoviral vector of claim 3, wherein the nucleic acid sequence coding for TNF further comprises a transcription-terminating region.
- 6. The adenoviral vector of claim 1, wherein the radiation inducible promoter is Egr-1.
- 7. The adenoviral vector of claim 2, wherein the adenoviral vector is deficient in the E3 region of the adenoviral genome.
- 8. The adenoviral vector of claim 7, wherein the adenoviral genome is deficient in the entire E3 region.
- 9. The adenoviral vector of claim 1, wherein the adenoviral genome is deficient in the entire coding region of the E4 region.
- 10. The adenoviral vector of claim 9, wherein the coding region of the E4 region of the adenoviral genome has been replaced with a spacer element having at least 15 base pairs.
- 11. The adenoviral vector of claim 10, wherein the spacer element is transcriptionally inert.
- 12. The adenoviral vector of claim 11, wherein the spacer element comprises a polyadenylation sequence that is non-native to the E4 region of the adenoviral genome.
- 13. The adenoviral vector of claim 12, wherein the spacer element comprises an SV40 polyadenylation sequence.
- 14. The adenoviral vector of claim 10, wherein the viral production level of the adenoviral vector is greater than the viral production level of the adenoviral vector without the spacer element.
- 15. The adenoviral vector of claim 1, wherein the adenoviral genome is a serotype-5 adenoviral genome.
- 16. A replication competent adenovirus-free stock of the adenoviral vector of claim 1.
- 17. A host cell comprising the adenoviral vector of claim 1.
- 18. An adenoviral vector comprising (a) an adenoviral genome deficient in the E4 region of the adenoviral genome, and optionally deficient in the E1 region, the E2A region, and/or the E3 region of the adenoviral genome, (b) a nucleic acid sequence coding for a TNF which is secreted from a cell infected with the adenoviral vector, and (c) a spacer element of at least 15 base pairs in the E4 region of the adenoviral genome.
- 19. The adenoviral vector of claim 18, wherein the adenoviral genome is deficient in the E1 region of the adenoviral genome.
- 20. The adenoviral vector of claim 18, wherein the spacer element is transcriptionally inert.
- 21. The adenoviral vector of claim 20, wherein the spacer element comprises a polyadenylation sequence that is non-native to the E4 region of the adenoviral genome.
- 22. The adenoviral vector of claim 21, wherein the spacer element comprises an SV40 polyadenylation sequence.
- 23. The adenoviral vector of claim 19, wherein the nucleic acid sequence coding for TNF is located in the E1 region of the adenoviral genome.
- 24. The adenoviral vector of claim 23, wherein the TNF is TNF-α.
- 25. The adenoviral vector of claim 23, wherein the nucleic acid sequence coding for TNF further comprises a transcription-terminating region.
- 26. The adenoviral vector of claim 18, wherein the adenoviral vector is deficient in the E3 region of the adenoviral genome.
- 27. The adenoviral vector of claim 26, wherein the adenoviral genome is deficient in the entire E3 region.
- 28. The adenoviral vector of claim 18, wherein the adenoviral genome is deficient in the entire coding region of the E4 region.
- 29. The adenoviral vector of claim 18, wherein the viral production level of the adenoviral vector is greater than the viral production level of the adenoviral vector without the spacer element.
- 30. The adenoviral vector of claim 18, wherein the adenoviral genome is a serotype-5 adenoviral genome.
- 31. A replication competent adenoviral-free stock of the adenoviral vector of claim 18.
- 32. A host cell comprising the adenoviral vector of claim 18.
- 33. A method of treating a tumor or cancer in a mammal comprising administering an anti-tumor or anti-cancer effective amount of the adenoviral vector of claim 1 directly to the tumor or cancer of the mammal.
- 34. The method of claim 33, further comprising the administration of radiation to the host.
- 35. The method of claim 34, wherein the radiation induces expression of the nucleic acid sequence coding for TNF to produce a therapeutic level of TNF in the host.
- 36. The method of claim 35, wherein the administration of radiation comprises the use of an internal source of radiation.
- 37. The method of claim 33, further comprising the administration of a TNF antagonist to the host.
- 38. The method of claim 37, wherein the TNF antagonist is at least one selected from the group comprising soluble TNF receptors and anti-TNF anti-bodies.
- 39. A method of treating a tumor or cancer in a mammal comprising administering an anti-tumor or anti-cancer effective amount of the adenoviral vector of claim 18 directly to the tumor or cancer of the mammal.
- 40. A method of producing an adenoviral vector comprising (a) providing an adenoviral genome that is deficient in the E4 region of the adenoviral genome, (b) inserting a nucleic acid sequence coding for TNF into the adenoviral genome, and (c) inserting a radiation-inducible promoter into the adenoviral genome such that it is operably linked to the nucleic acid sequence coding for TNF.
- 41. The method of claim 40, wherein the adenoviral genome comprises a spacer element having at least 15 base pairs in the E4 region of the adenoviral genome.
- 42. The method of claim 40, wherein the radiation inducible promoter is Egr-1.
- 43. The adenoviral vector of claim 1, wherein the nucleic acid sequence encodes a TNF which is secreted from a cell infected with the adenoviral vector.
- 44. The adenoviral vector of claim 1, wherein the nucleic acid sequence coding for TNF comprises SEQ ID NO: 2.
- 45. The adenoviral vector of claim 18, wherein the nucleic acid sequence coding for a TNF which is secreted from a cell infected with the adenoviral vector comprises SEQ ID NO: 2.
- 46. The method of claim 33, wherein the adenoviral vector is administered to a tumor in a mammal.
- 47. The method of claim 46, wherein the mammal is a human.
- 48. The method of claim 39, wherein the adenoviral vector is administered to a tumor in a mammal.
- 49. The method of claim 48, wherein the mammal is a human.
- 50. A pharmaceutical composition comprising the adenoviral vector of claim 1 and a pharmaceutically acceptable carrier, wherein the pharmaceutical composition does not contain replication-competent adenoviruses.
- 51. A pharmaceutical composition comprising the adenoviral vector of claim 18 and a pharmaceutically acceptable carrier, wherein the pharmaceutical composition does not contain replication-competent adenoviruses.
CROSS-REFERENCE TO RELATED PATENT APPLICATIONS
[0001] This patent application is a divisional of U.S. patent application Ser. No. 09/604,694, filed Jun. 27, 2000.
Continuations (1)
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Number |
Date |
Country |
Parent |
09604694 |
Jun 2000 |
US |
Child |
10424638 |
Apr 2003 |
US |