DNA vaccines against tumor growth and methods of use thereof

• US 20100136058A1
• Filed: 08/04/2009
• Published: 06/03/2010
• Est. Priority Date: 10/08/2003
• Status: Active Grant

• Current Assignee: The Scripps Research Institute
• Original Assignee: The Scripps Research Institute
• Inventors: Xiang, Rong, Luo, Yunping, Reisfeld, Ralph A.
• Granted Patent: US 8,053,421 B2
• Time in Patent Office: Days
• US Class Current: 424/246.100
• CPC Class Codes: A61K 2039/522 avirulent or attenuated
• CPC Class Codes: A61K 2039/523 expressing foreign proteins
• CPC Class Codes: A61K 2039/53 DNA (RNA) vaccination
• CPC Class Codes: A61K 2039/55527 Interleukins
• CPC Class Codes: A61K 2039/6031 Proteins
• CPC Class Codes: A61K 39/0011 Cancer antigens
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4613 Natural-killer cells [NK or NK-T]
• CPC Class Codes: A61K 39/464402 Receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: A61K 48/0008 characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition
• CPC Class Codes: A61K 48/005 characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
• CPC Class Codes: C12N 2510/02 Cells for production
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

A DNA vaccine suitable for eliciting an immune response against cancer cells comprises a polynucleotide construct operably encoding an a Fra-1 protein, such as a polyubiquitinated human Fra-1 protein, and IL-18, such as human IL-18, in a pharmaceutically acceptable carrier. In a preferred embodiment, the polynucleotide construct is operably incorporated in an attenuated bacterial vector, such as an attenuated Salmonella typhimurium, particularly a doubly attenuated aroA^- dam^- S. typhimurium. Transformed host cells, methods of inhibiting tumor growth, of vaccinating a patient against cancer, and of delivering genetic material to a mammalian cell in vivo are also described.

Claims

1. A DNA vaccine suitable for eliciting an immune response against cancer cells that overexpress Fra-1, the vaccine comprising a polynucleotide construct operably encoding a Fra-1 protein and IL-18 in a pharmaceutically acceptable carrier.

2. The DNA vaccine of claim 1 wherein the polynucleotide construct is operably incorporated in a vector.

3. The DNA vaccine of claim 2 wherein the vector is an attenuated bacterial vector.

4. The DNA vaccine of claim 3 wherein the attenuated bacterial vector is selected from the group consisting of attenuated Salmonella typhimurium, Salmonella typhi, Shigella, Bacillus, Lactobacillus, BCG, Escherichia coli, Vibrio cholerae, and Campylobacter.

5. The DNA vaccine of claim 1 wherein the polynucleotide construct encodes a Fra-1 protein having an amino acid sequence selected from the group consisting of SEQ ID NO:

2 and SEQ ID NO;

4.

6. The DNA vaccine of claim 1 wherein the polynucleotide construct encodes IL-18 having an amino acid sequence selected from the group consisting of SEQ ID NO:

6 and SEQ ID NO;

8.

7. The DNA vaccine of claim 1 wherein the polynucleotide construct comprises a polynucleotide encoding a human or murine Fra-1 protein, and having a nucleic acid sequence selected from the group consisting of SEQ ID NO:

1 and SEQ ID NO;

3.

8. The DNA vaccine of claim 1 wherein the polynucleotide construct comprises a polynucleotide encoding a human or murine IL-18, and having a nucleic acid sequence selected from the group consisting of SEQ ID NO:

5 and SEQ ID NO;

7.

9. The DNA vaccine of claim 1 wherein the polynucleotide construct further encodes IL-12.

10. The DNA vaccine of claim 1 wherein the polynucleotide construct comprises an Fra-1 polynucleotide having a nucleic acid sequence selected from the group consisting of SEQ ID NO:

1, and SEQ ID NO;

3 and an IL-18 polynucleotide having a nucleic acid sequence selected from the group consisting of SEQ ID NO;

5, and SEQ ID NO;

7.

11. A method of inhibiting tumor growth in a mammal having a Fra-1 overexpressing tumor and comprising the step of administering to the mammal an effective immunological response eliciting amount of a DNA vaccine comprising a polynucleotide construct operably encoding a Fra-1 protein and IL-18 in a pharmaceutically acceptable carrier, whereby the mammal exhibits an immune response elicited by vaccine and specific to tumor cells.

12. The method of claim 11 wherein the polynucleotide construct encodes IL-18 having an amino acid sequence selected from the group consisting of SEQ ID NO:

6 and SEQ ID NO;

8.

13. The method of claim 11 wherein the polynucleotide construct comprises a polynucleotide encoding a human or murine IL-18, and having a nucleic acid sequence selected from the group consisting of SEQ ID NO:

5 and SEQ ID NO;

7.

14. The method of claim 11 wherein the mammal is a human.

15. The method of claim 11 wherein the polynucleotide construct is operably incorporated in an attenuated bacterial vector.

16. The method of claim 15 wherein the attenuated bacterial vector is selected from attenuated Salmonella typhimurium, Salmonella typhi, Shigella, Bacillus, Lactobacillus, BCG, Escherichia coli, Vibrio cholerae, and Campylobacter.

17. The method of claim 15 wherein the vaccine is administered orally.

18. A transformed host cell transfected with a polynucleotide construct operably encoding a Fra-1 protein and IL-18.

19. A method of vaccinating a mammal against a Fra-1 overexpressing cancer, the method comprising the step of administering to the mammal an effective immunological response eliciting amount of a DNA vaccine comprising a polynucleotide construct operably encoding a Fra-1 protein and IL-18 in a pharmaceutically acceptable carrier.

20. The method of claim 19 wherein the vaccine is administered orally.

21. A method of delivery of genetic material to a mammalian cell in vivo comprising orally administering to a mammal a polynucleotide construct operably encoding a therapeutically useful gene product.

22. The method of claim 21 wherein the therapeutically useful gene product is a tumor antigen capable of eliciting an immune response in the mammal against tumor cells.

23. The method of claim 21 wherein the therapeutically useful gene product is an immune stimulating molecule capable of stimulating the immune system of the mammal.

24. The method of claim 21 wherein the therapeutically useful gene product comprises a tumor antigen capable of eliciting an immune response in the mammal against tumor cells and an immune stimulating molecule capable of stimulating the immune system of the mammal.

25. The method of claim 23 wherein the tumor antigen is a Fra-1 protein and the immune stimulating molecule is IL-18.