Oncolytic adenovirus encoding a B7 protein
The present disclosure provides a replication competent oncolytic adenovirus with selectivity for cancer cells, wherein the adenovirus comprises a transgene under the control of a promoter endogenous to the virus, wherein the transgene comprises a DNA sequence encoding a B7 protein or an active fragment thereof, compositions comprising same, methods of generating the viruses, and use of the viruses and compositions in treatment, particularly in the treatment of cancer.
1. A replication competent oncolytic group B adenovirus with selectivity for cancer cells, wherein the adenovirus comprises a transgene under the control of a promoter endogenous to the virus, wherein the transgene comprises a DNA sequence encoding a B7 protein or an active fragment thereof, and
said transgene is located between the stop codon-polyA recognition site of the adenovirus L5 gene and the stop codon-polyA recognition site of the adenovirus E4 gene,
wherein said transgene is under the control of the endogenous major late promoter.
2. A replication competent oncolytic virus according to claim 1 , wherein the B7 protein or active fragment thereof is independently selected from the group comprising B7-1, B7-2, B7-DC, B7-H1, B7-H2, B7-H3, B7-H4, B7-H5 and B7-H6.
3. A replication competent oncolytic virus according to claim 2 , wherein the B7 protein or active fragment thereof is B7-1.
4. A replication competent oncolytic virus according to claim 1 , wherein the virus is a chimeric virus.
5. A replication competent oncolytic virus according to claim 4 , wherein the virus backbone is enadenotucirev.
6. A replication competent oncolytic virus according to claim 5 , wherein the virus has a formula (I):
5′ITR-B 1 -B A -B 2 -B X -B B -B Y -B 3 -3′ITR (I)
wherein:
B 1 comprises: E1A, E1B or E1A-E1B;
B A is E2B-L1-L2-L3-E2A-L4;
B 2 is absent or comprises E3 or a transgene under an endogenous or exogenous promoter;
B X is absent or a DNA sequence comprising: a restriction site, one or more transgenes or both;
B B comprises L5;
B Y comprises a transgene encoding a B7 protein or an active fragment thereof; and
B 3 is absent or comprises E4.
7. A replication competent oncolytic virus according to claim 1 , wherein the B7 protein or active fragment thereof comprises a transmembrane sequence selected from the group comprising a transmembrane domain from a PDGF receptor, or a GPI anchor suitable for anchoring the protein or fragment in a cell membrane.
8. A replication competent oncolytic virus according to claim 1 , which further comprises a second transgene encoding a polypeptide selected from the group comprising a cytokine, a chemokine, an antagonistic antibody molecule or fragment thereof, and an agonistic antibody molecule or fragment thereof.
9. A replication competent oncolytic virus according to claim 1 , which comprises a second and third transgene encoding two different polypeptides selected from the group comprising a cytokine, a chemokine, an antagonistic antibody molecule or fragment thereof, and an agonistic antibody molecule or fragment thereof.
10. A replication competent oncolytic virus according to claim 9 , wherein the second or third transgene encodes a cytokine selected from the group comprising IL-2, IFN-alpha, IFN-beta, IFN-gamma, Flt3 ligand, GM-CSF, IL-15, and IL-12.
11. A replication competent oncolytic virus according to claim 9 , wherein the second or third transgene encodes a chemokine selected from the group comprising MIP-1 alpha, IL-8, CCL5, CCL17, CCL20, CCL22, CXCL9, CXCL10, CXCL11, CXCL13, CXCL12, CCL2, CCL19 and CCL21.
12. A replication competent oncolytic virus according to claim 8 , wherein a cytokine and a chemokine combination is selected from the group comprising Mip1α and Flt3 ligand, and Mip1α and IFNα.
13. A replication competent oncolytic virus according to claim 1 , wherein the virus encodes an antibody molecule or fragment thereof.
14. A replication competent oncolytic virus according to claim 13 , wherein the antibody or antibody fragment molecule comprises a transmembrane sequence or GPI anchor such that it is a cell membrane-anchored form or a transmembrane domain.
15. A replication competent oncolytic virus according to claim 13 , wherein the antibody or antibody fragment molecule comprises an anti-human CD3 antigen binding domain.
16. A replication competent oncolytic virus according to claim 13 , wherein the antibody molecule is an inhibitor selected from the group comprising an anti-VEGF antibody molecule, and an anti-CTLA-4 antibody molecule.
17. A replication competent oncolytic virus according to claim 13 , wherein the antibody molecule is an agonist selected from the group comprising CD40, GITR, OX40, CD27 and 4-1BB.
18. A pharmaceutical composition comprising a replication competent oncolytic adenovirus according to claim 1 , and a diluent or carrier.
19. A method of treating a patient comprising administering a therapeutically effective amount of a replication competent adenovirus according to claim 1 .
20. A method of treating a patient comprising administering a therapeutically effective amount of a composition according to claim 18 .
21. A method of generating a replication competent adenovirus according to claim 1 by replication in a host cell.