Recombinant adenoviral vector and methods of use

• US 6,210,939 B1
• Filed: 10/25/1994
• Issued: 04/03/2001
• Est. Priority Date: 10/25/1993
• Status: Expired due to Term

• Current Assignee: Canji Inc. (Merck & Co., Inc.)
• Original Assignee: Canji Inc. (Merck & Co., Inc.)
• Inventors: Maneval, Daniel C., Wills, Ken N., Gregory, Richard J.
• Primary Examiner(s): Guzo, David
• Application Number:
  US08/328,673
• Time in Patent Office: 2,352 Days
• Field of Search: 435/320.1, 435/172.3, 435/69.1, 435/172.1, 435/325, 435/363, 435/366, 435/370, 435/371, 435/252.3, 424/93.2, 424/93.6, 514/44, 935/32
• US Class Current: 435/456
• 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: C07K 14/4746 p53
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2710/10343 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2830/008 cell type or tissue specific enhancer/promoter combination
• CPC Class Codes: C12N 2830/85 mammalian
• 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

This invention provides a recombinant adenovirus expression vector characterized by the partial or total deletion of the adenoviral protein IX DNA and having a gene encoding a foreign protein or a functional fragment or mutant thereof. Transformed host cells and a method of producing recombinant proteins and gene therapy also are included within the scope of this invention. Thus, for example, the adenoviral vector of this invention can contain a foreign gene for the expression of a protein effective in regulating the cell cycle, such as p53, Rb, or mitosin, or in inducing cell death, such as the conditional suicide gene thymidine kinase. (The latter must be used in conjunction with a thymidine kinase metabolite in order to be effective).

Claims

1. A recombinant adenovirus expression vector comprising

(a) an insert of exogenous DNA comprising a gene encoding a foreign protein and

(b) adenovirus DNA which has sustained a deletion beginning at nucleotide 357 and ending at nucleotide 4020 to 4050.

2. The recombinant adenovirus expression vector of claim 1, wherein the total length of the recombinant adenovirus expression vector is less than 35 KB.

3. The recombinant adenovirus expression vector of claim 1, wherein the insert is less than 4.5 KB.

4. The recombinant adenovirus expression vector of claim 1, wherein the insert further comprises an exogenous polyadenylation signal.

5. The recombinant adenovirus expression vector of claim 1, wherein the foreign protein is a tumor suppressor protein.

6. The recombinant adenovirus expression vector of claim 1, wherein the gene encoding the foreign protein is expressed under control of a cytomegalovirus (CMV) promoter.

7. The recombinant adenovirus expression vector of claim 1, wherein the CMV promoter is the CMV immediate early promoter.

8. The recombinant adenovirus expression vector of claim 1, wherein the gene encoding the foreign protein is expressed under control of the adenovirus 2 major late promoter.

9. The recombinant adenovirus expression vector of claim 1, wherein the vector is A/C/N/53.

10. The recombinant adenovirus expression vector of claim 1, wherein the vector is A/M/N/53.

11. The recombinant adenovirus expression vector of claim 1, wherein the adenovirus is a Group C adenovirus selected from a serotype 1, 2, 5 or 6.

12. A transformed host cell comprising the recombinant adenovirus expression vector of claim 1.

13. The transformed host cell of claim 1, wherein the host cell is a procaryotic or eucaryotic cell.

14. A method for transforming a pathologic hyperproliferative mammalian cell comprising contacting the cell with the expression vector of claim 1.

15. The recombinant adenovirus expression vector of claim 5, wherein the tumor suppressor protein is p53.

16. The recombinant adenovirus expression vector of claim 5, wherein the tumor suppressor protein is RB56.

17. A recombinant adenovirus expression vector comprising

(a) an insert of exogenous DNA comprising a gene encoding a foreign protein;

and

(b) adenovirus DNA in which adenovirus polynucleotide sequences beginning at nucleotide 360 and ending at nucleotide 4020 to 4050 are replaced with a non-adenoviral polynucleotide sequence.

18. The recombinant adenovirus expression vector of claim 17, wherein the vector is A/C/N/53.

19. The recombinant adenovirus expression vector of claim 17, wherein the foreign protein is a tumor suppressor protein.

20. The recombinant adenovirus expression vector of claim 17, wherein the gene encoding the foreign protein is expressed under control of a cytomegalovirus (CMV) promoter.

21. The recombinant adenovirus expression vector of claim 17, wherein the gene encoding the foreign protein is expressed under control of the adenovirus 2 major late promoter.

22. The recombinant adenovirus expression vector of claim 19, wherein the tumor suppressor protein is p53.

23. The recombinant adenovirus expression vector of claim 19, wherein the tumor suppressor protein is RB56.

24. The recombinant adenovirus expression vector of claim 20, wherein the CMV promoter is the CMV immediate early promoter.

25. A recombinant adenovirus expression vector, wherein the vector is A/C/N/53.