Method of sensitizing tumor cells with adenovirus E1A

• US 6,100,243 A
• Filed: 05/09/1997
• Issued: 08/08/2000
• Est. Priority Date: 09/06/1994
• Status: Expired due to Fees

• Current Assignee: La Jolla Cancer Research Foundation
• Original Assignee: La Jolla Cancer Research Foundation
• Inventors: Frisch, Steven M.
• Primary Examiner(s): LeGuyader, John L.
• Assistant Examiner(s): Nguyen, Dave Trong
• Application Number:
  US08/853,831
• Time in Patent Office: 1,187 Days
• Field of Search: 514/44, 424/93.21, 536/23.1, 435/69.1, 435/320.1, 435/455, 435/458
• US Class Current: 514/44.00R
• 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 41/0038 Radiosensitizing, i.e. administration of pharmaceutical agents that enhance the effect of radiotherapy radiotherapy per se A61N5/10
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C12N 2710/10322 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Abstract

The present invention is directed to methods of sensitizing a human tumor cell with adenovirus E1A. The methods involve treating a human tumor cell by, first, introducing into the tumor cell nucleic acid encoding a polypeptide having adenovirus E1A activity, expressing the E1A active polypeptide in the cell, and then either contacting the E1A expressing tumor cell with a chemotherapeutic agent or irradiating the E1A-expressing tumor cell. The invention also provides methods of enhancing a subject'"'"'s response to chemotherapy or irradiation by introducing into a subject'"'"'s tumor cells nucleic acid encoding a polypeptide having adenovirus E1A activity, expressing the E1A active polypeptide in the cells and finally, administering either a chemotherapeutic agent or irradiation. The invention also provides a method of treating cancer.

Claims

1. A method of reducing viability of a human tumor cell in a subject, comprising the steps of:

a. introducing into a human tumor cell in a subject nucleic acid encoding a polypeptide having adenovirus E1A activity;

b. expressing said polypeptide in an effective amount in the tumor cell, thereby enhancing sensitivity of the tumor cell expressing said polypeptide to a chemotherapeutic agent; and

c. contacting the tumor cell with said chemotherapeutic agent

wherein a nucleic acid encoding functional p53 is not introduced into the tumor cell, said enhanced sensitivity is independent or p53 and viability of the tumor cell is reduced.

2. The method of claim 1, wherein said human tumor cell does not express functional p53.

3. The method of claim 1, wherein the nucleic acid encodes adenovirus E1A polypeptide (SEQ ID NO:

2).

4. The method of claim 1, wherein the nucleic acid is introduced into the tumor cell by contacting the cell with a suitable retroviral vector containing the nucleic acid.

5. The method of claim 1, wherein the chemotherapeutic agent is selected from the group consisting of etoposide, adriamycin, amsacrine, actinomycin D, VP16, camptothecin, colchicine, taxol, cisplatinum, vincristine, vinblastine, and methotrexate.

6. A method of reducing viability of a human tumor cell in a subject, comprising the steps of:

a. introducing into a human tumor cell in a subject nucleic acid encoding a polypeptide having adenovirus E1A activity;

b. expressing said polypeptide in an effective amount in the tumor cell, thereby enhancing sensitivity of the tumor cell expressing said polypeptide to irradiation; and

c. irradiating the tumor cell;

wherein a nucleic acid encoding functional p53 is not introduced into the tumor cell, said enhanced sensitivity is independent of p53 and viability of the tumor cell is reduced.

7. The method of claim 6, wherein said human tumor cell does not express functional p53.

8. The method of claim 6, wherein the nucleic acid encodes adenovirus E1A polypeptide (SEQ ID NO:

2).

9. The method of claim 8, wherein the nucleic acid is introduced into the tumor cell by contacting the cell with a suitable retroviral vector containing the nucleic acid.

10. A method of increasing susceptibility to chemotherapy in a subject, comprising the steps of:

a. introducing into a subject'"'"'s tumor cells nucleic acid encoding a polypeptide having adenovirus E1A activity;

b. expressing said polypeptide in an effective amount in said tumor cells, thereby increasing susceptibility of said tumor cells expressing said polypeptide to said chemotherapeutic agent; and

c. administering said chemotherapeutic agent to said subject;

wherein a nucleic acid encoding functional p53 is not introduced into said tumor cells and said icreased susceptibility is independent of p53.

11. The method of claim 10, wherein the introducing step (a) is by direct injection into said subject.

12. The method of claim 10, wherein said tumor cells do not express functional p53.

13. The method of claim 10, wherein the nucleic acid encodes adenovirus E1A polypeptide (SEQ ID NO:

2).

14. A method of increasing susceptibility to irradiation in a subject, comprising the steps of:

a. introducing into a subject'"'"'s tumor cells nucleic acid encoding a polypeptide having adenovirus E1A activity;

b. expressing said polypeptide in an effective amount in said tumor cells, thereby increasing susceptibility of said tumor cells expressing said polypeptide to irradiation; and

c. administering irradiation to said subject;

wherein a nucleic acid encoding functional p53 is not introduced into said tumor cells and said increased susceptibility is independent of p53.

15. The method of claim 14, wherein the introducing step (a) is by direct injection into said subject.

16. The method of claim 14, wherein said tumor cells do not express functional p53.

17. The method of claim 14, wherein the nucleic acid encodes adenovirus E1A polypeptide (SEQ ID NO:

2).

18. A method of inactivating a tumor in a subject, comprising the steps of:

a. introducing into a subject'"'"'s tumor cells nucleic acid encoding a polypeptide having adenovirus E1A activity;

b. expressing said polypeptide in an effective amount in said tumor cells, thereby enhancing sensitivity of said tumor to a chemotherapeutic agent; and

c. contacting said tumor with said chemotherapeutic agent;

wherein a nucleic acid encoding functional p53 is not introduced into said tumor cells, said enhanced sensitivity is independent of p53 and said tumor is inactivated.

20. The method of claim 18, wherein said tumor cells do not express functional p53.

21. The method of claim 18, wherein the nucleic acid encodes adenovirus E1A polypeptide (SEQ ID NO:

2).

22. The method of claim 18, wherein the nucleic acid is introduced into the tumor cells by contacting the cells with a suitable retroviral vector containing the nucleic acid.

23. The method of claim 18, wherein the chemotherapeutic agent is selected from the group consisting of etoposide, adriamycin, amsacrine, actinomycin D, VP16, camptothecin, colchicine, taxol, cisplatinum, vincristine, vinblastine, and methotrexate.

19. A method of inactivating a tumor in a subject, comprising the steps of:

a. introducing into a subject'"'"'s tumor cells nucleic acid encoding a polypeptide having adenovirus E1A activity;

b. expressing said polypeptide in an effective amount in said tumor cells, thereby enhancing sensitivity of said tumor to irradiation; and

c. irradiating said tumor;

wherein a nucleic acid encodina functional p53 is not introduced into said tumor cells, said enhanced sensitivity is independent of p53 and said tumor is inactivated.

24. The method of claim 19, wherein said tumor cells do not express functional p53.

25. The method of claim 19, wherein the nucleic acid encodes adenovirus E1A polypeptide (SEQ ID NO:

2).

26. The method of claim 19, wherein the nucleic acid is introduced into the tumor cells by contacting the cells with a suitable retroviral vector containing the nucleic acid.