Stocks of recombinant, replication-deficient adenovirus free of replication-competent adenovirus

• US 5,994,106 A
• Filed: 11/26/1996
• Issued: 11/30/1999
• Est. Priority Date: 06/10/1994
• Status: Expired due to Fees

• Current Assignee: GenVec, Inc. (Precigen, Inc.)
• Original Assignee: GenVec, Inc. (Precigen, Inc.)
• Inventors: Brough, Douglas E., Kovesdi, Imre, McVey, Duncan L., Bruder, Joseph T., Lizonova, Alena
• Primary Examiner(s): PRIEBE, SCOTT DAVID
• Application Number:
  US08/757,023
• Time in Patent Office: 1,099 Days
• Field of Search: 435/69.1, 435/91.1, 435/172.1, 435/172.3, 435/375, 435/366, 435/320.1, 435/91.4, 435/455, 435/456, 435/325, 514/44, 424/199.1, 536/23.72, 536/24.1
• US Class Current: 435/91.4
• CPC Class Codes: A61K 2039/5256 expressing foreign proteins
• 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 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C07K 14/4712 Cystic fibrosis
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2710/10322 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2710/10343 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2710/10352 relating to complementing cells and packaging systems for producing virus or viral particles
• CPC Class Codes: C12N 2810/60 from viruses
• CPC Class Codes: C12N 2810/6081 rhabdoviridae, e.g. VSV
• CPC Class Codes: C12N 2830/002 inducible enhancer/promoter combination, e.g. hypoxia, iron, transcription factor
• CPC Class Codes: C12N 2830/85 mammalian
• CPC Class Codes: C12N 2840/20 translation of more than one cistron
• CPC Class Codes: C12N 2840/203 having an IRES
• CPC Class Codes: C12N 2840/44 being a specific part of the splice mechanism, e.g. donor, acceptor
• 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
• CPC Class Codes: Y10S 977/799 Containing biological material

Abstract

The present invention provides multiply deficient adenoviral vectors and complementing cell lines. Also provided are recombinants of the multiply deficient adenoviral vectors and a therapeutic method, particularly relating to gene therapy, vaccination, and the like, involving the use of such recombinants.

Claims

1. A replication-competent adenovirus-free stock of a recombinant adenoviral vector, wherein said adenoviral vector is deficient in one or more essential gene functions of one or more regions of the adenoviral genome selected from the group consisting of the E1, E2A , and E4 regions of the adenoviral genome.

2. The stock of claim 1, wherein said adenoviral vector is deficient in one or more essential gene functions of E1.

3. The stock of claim 1, wherein said adenoviral vector is deficient in one or more essential gene functions of E2A .

4. The stock of claim 1, wherein said adenoviral vector is deficient in one or more essential gene functions of E4.

5. The stock of claim 1, wherein said adenoviral vector is deficient in two or more essential gene functions.

6. The stock of claim 5, wherein said adenoviral vector is deficient in one or more essential gene functions of each of the E1 and E2A regions of the adenoviral genome.

7. The stock of claim 5, wherein said adenoviral vector is deficient in one or more essential gene functions of each of the E1 and E4 regions of the adenoviral genome.

8. The stock of claim 5, wherein said adenoviral vector is deficient in one or more essential gene functions of each of the E2A and E4 regions of the adenoviral genome.

9. The stock of claim 1, wherein said adenoviral vector is deficient in three essential gene functions.

10. The stock of claim 9, wherein said adenoviral vector is deficient in one or more essential gene functions of each of the E1, E2A , and E4 regions of the adenoviral genome.

11. The stock of claim 1, wherein said adenoviral vector is prepared in a cell that complements in trans the deficient essential gene functions of said adenoviral vector, wherein the genome of said cell line is free of overlapping sequences with said adenoviral vector that are sufficient to mediate a recombination event resulting in a replication competent adenoviral vector.

12. The stock of claim 2, wherein said adenoviral vector is prepared in a cell that complements in trans the deficient essential gene functions of said adenoviral vector, wherein the genome of said cell line is free of overlapping sequences with said adenoviral vector that are sufficient to mediate a recombination event resulting in a replication competent adenoviral vector.

13. The stock of claim 3, wherein said adenoviral vector is prepared in a cell that complements in trans the deficient essential gene functions of said adenoviral vector, wherein the genome of said cell line is free of overlapping sequences with said adenoviral vector that are sufficient to mediate a recombination event resulting in a replication competent adenoviral vector.

14. The stock of claim 4, wherein said adenoviral vector is prepared in a cell that complements in trans the deficient essential gene functions of said vector, wherein the genome of said cell line is free of overlapping sequences with said vector that are sufficient to mediate a recombination event resulting in a replication competent adenoviral vector.

15. The stock of claim 5, wherein said adenoviral vector is prepared in a cell that complements in trans the deficient essential gene functions of said adenoviral vector, wherein the genome of said cell line is free of overlapping sequences with said adenoviral vector that are sufficient to mediate a recombination event resulting in a replication competent adenoviral vector.

16. The stock of claim 6, wherein said adenoviral vector is prepared in a cell that complements in trans the deficient essential gene functions of said adenoviral vector, wherein the genome of said cell line is free of overlapping sequences with said adenoviral vector that are sufficient to mediate a recombination event resulting in a replication competent adenoviral vector.

17. The stock of claim 7, wherein said adenoviral vector is prepared in a cell that complements in trans the deficient essential gene functions of said adenoviral vector, wherein the genome of said cell line is free of overlapping sequences with said adenoviral vector that are sufficient to mediate a recombination event resulting in a replication competent adenoviral vector.

18. The stock of claim 8, wherein said adenoviral vector is prepared in a cell that complements in trans the deficient essential gene functions of said adenoviral vector, wherein the genome of said cell line is free of overlapping sequences with said adenoviral vector that are sufficient to mediate a recombination event resulting in a replication competent adenoviral vector.

19. The stock of claim 9, wherein said adenoviral vector is prepared in a cell that complements in trans the deficient essential gene functions of said adenoviral vector, wherein the genome of said cell line is free of overlapping sequences with said adenoviral vector that are sufficient to mediate a recombination event resulting in a replication competent adenoviral vector.

20. The stock of claim 10, wherein said adenoviral vector is prepared in a cell that complements in trans the deficient essential gene functions of said adenoviral vector, wherein the genome of said cell line is free of overlapping sequences with said adenoviral vector that are sufficient to mediate a recombination event resulting in a replication competent adenoviral vector.

21. A system comprising (i) a cell having a cellular genome comprising at least one DNA sequence, which is operably linked to an inducible promoter or a repressible promoter, and which complements in trans for a deficiency in one or more essential gene functions of the E1 and E2A regions of an adenoviral genome, and (ii) an adenoviral vector comprising an adenoviral genome having a deficiency in one or more essential gene functions of the E1 and E2A regions of said adenoviral genome, wherein there is insufficient overlap between said cellular genorne and said adenoviral genome to mediate a recombination event sufficient to result in a replication-competent adenovirus.

22. A system comprising (i) a cell having a cellular genome comprising at least one DNA sequence, which is operably linked to an inducible promoter or a repressible promoter, and which complements in trans for a deficiency in one or more essential gene functions of the E1, E2A and E4 regions of an adenoviral genome, and (ii) an adenoviral vector comprising an adenoviral genome having a deficiency in one or more essential gene functions of the E1, E2A and E4 regions of said adenoviral genome, wherein there is insufficient overlap between said cellular genome and said adenoviral genome to mediate a recombination event sufficient to result in a replication-competent adenovirus.

23. A method of propagating an adenoviral vector that is deficient in one or more essential gene functions of one or more regions of an adenoviral genome, which method comprises propagating an adenoviral vector comprising an adenoviral genome having a deficiency in one or more essential gene functions of the E1 and E2A regions in a cell having a cellular genome comprising at least one DNA sequence that complements in trans said deficiency, wherein there is insufficient overlap between said cellular genome and said adenoviral genome to mediate a recombination event resulting in a replication-competent adenoviral vector.

24. A method of propagating an adenoviral vector that is deficient in one or more essential gene functions of one or more regions of an adenoviral genome, which method comprises propagating an adenoviral vector comprising an adenoviral genome having a deficiency in one or more essential gene finctions of the E1, E2A and E4 regions in a cell having a cellular genome comprising at least one DNA sequence that complements in trans said deficiency, wherein there is insufficient overlap between said cellular genome and said adenoviral genome to mediate a recombination event resulting in a replication-competent adenoviral vector.