Oncolytic virus strain

1. A herpes simplex virus (HSV) which expresses at least three heterologous genes, wherein each of the three heterologous genes is terminated by a different poly adenylation sequence selected from the BGH, SV40, HGH, and RBG poly adenylation sequences.

2. The virus of claim 1 , which is a HSV1.

3. The virus of claim 1 , which is a modified clinical isolate.

4. The virus of claim 3 wherein the clinical isolate is HSV1:

strain RH018A having the accession number ECACC 16121904;

strain RH004A having the accession number ECACC 16121902;

strain RH031A having the accession number ECACC 16121907;

strain RH040B having the accession number ECACC 16121908;

strain RH015A having the accession number ECACC 16121903;

strain RH021A having the accession number ECACC 16121905;

strain RH023A having the accession number ECACC 16121906;

or

strain RH047A having the accession number ECACC 16121909.

5. The virus of claim 1 , wherein the at least three heterologous genes comprise:

(a) one or more immune stimulatory molecule-encoding genes;

and/or

(b) one or more fusogenic protein-encoding genes.

6. The virus of claim 5 , wherein:

(a) the fusogenic protein is selected from the group consisting of vesicular stomatitis virus (VSV) G-protein, syncitin-1, syncitin-2, simian virus 5 (SV5) F-protein, measles virus (MV) H-protein, MV F-protein, respiratory syncytial virus (RSV) F-protein, and a glycoprotein from gibbon ape leukemia virus (GALV), murine leukemia virus (MLV), Mason-Pfizer monkey virus (MPMV), or equine infectious anaemia virus (EIAV) from which the R peptide has been deleted; and/or

(b) the immune stimulatory molecule is GM-CSF, IL-2, IL-12, IL-15, IL-18, IL-21, IL-24, a type I interferon, interferon gamma, a type III interferon, TNF alpha, an antagonist of TGF beta, an immune checkpoint antagonist, or an agonist of an immune potentiating pathway including an agonist of CD40, ICOS, GITR, 4-1-BB, OX40, or flt3.

7. The virus of claim 6 , wherein:

(a) the fusogenic protein is the glycoprotein from gibbon ape leukemia virus (GALV) and has the R transmembrane peptide mutated or removed (GALV-R-);

(b) the agonist of an immune potentiating pathway is CD40 ligand (CD40L), ICOS ligand, GITR ligand, 4-1-BB ligand, OX40 ligand, or flt3 ligand; and/or

(c) the immune checkpoint antagonist is a CTLA-4 inhibitor.

8. The virus of claim 7 , wherein the at least three heterologous genes comprise GALV-R- and one or more of GM-CSF, a CTLA-4 inhibitor, an agonist of CD40, an agonist of GITR, an agonist of OX40, and an agonist of ICOS.

9. The virus of claim 8 , wherein the CTLA-4 inhibitor is an anti-CTLA4 antibody, the agonist of CD40 is CD40L, the agonist of CD40 is 41BBL, the agonist of GITR is GITRL, the agonist of ICOS is ICOSL, and/or the agonist of OX40 is OX4OL.

10. The virus of claim 8 , wherein the HSV is a modified HSV1 strain RH018A having the accession number ECACC 16121904.

11. The virus of claim 1 , wherein the virus:

(a) does not express functional ICP34.5;

(b) does not express functional ICP47; and/or

(c) expresses the US11 gene as an immediate early gene.

12. The virus of claim 1 , which expresses four heterologous genes driven by each of the CMV promoter, the RSV promoter, the SV40 promoter and a retroviral LTR promoter, respectively, and/or terminated by each of the BGH, SV40, HGH and RBG poly adenylation sequences, respectively.

13. A pharmaceutical composition comprising a virus according to claim 1 and a pharmaceutically acceptable carrier or diluent.

14. A product of manufacture comprising a virus according to claim 1 in a sterile vial, ampoule or syringe.

15. A method of treating cancer, which comprises administering a therapeutically effective amount of the virus of claim 1 to a patient in need thereof.

16. A method according to claim 15 , which further comprises administering a therapeutically effective amount of a further anti-cancer agent to a patient in need thereof.

17. A method according to claim 16 , wherein the further anti-cancer agent is selected from an antagonist of an immune co-inhibitory pathway, an agonist of an immune co-stimulatory pathway, radiation and/or chemotherapy, an agent that targets a specific genetic mutation which occurs in tumors, an agent intended to induce an immune response to one or more tumor antigen(s) or neoantigen(s), a cellular product of T cells or NK cells, an agent intended to stimulate the STING, cGAS, TLR or other innate immune response and/or inflammatory pathway, a second virus optionally an oncolytic virus, an inhibitor of the indoleamine 2,3-dioxygenase (IDO) pathway, and combinations thereof.

18. A method according to claim 17 , wherein the antagonist of an immune co-inhibitory pathway is a CTLA-4 inhibitor, a PD-1 inhibitor, a PD-L1 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor, a VISTA inhibitor, aCSF1R inhibitor, an IDO inhibitor, a CEACAM1 inhibitor, a KIR inhibitor, a SLAMF7 inhibitor or a CD47 inhibitor, and/or the agonist of an immune co-stimulatory pathway is a GITR agonist, a 4-1-BB agonist, an OX40 agonist, a CD40 agonist or an ICOS agonist.

19. A method according to claim 16 , wherein the further anti-cancer agent comprises an antibody.

20. A method according to claim 16 , wherein the virus and the further anti-cancer agent(s) are administered separately or concurrently.

21. A method according to claim 15 , wherein the cancer is a solid tumor.

22. The virus of claim 1 , wherein each of the three heterologous genes is driven by a different promoter selected from the CMV promoter, the RSV promoter, the SV40 promoter, and a retroviral LTR promoter.

23. The virus of claim 22 , which is a HSV 1.

24. The virus of claim 22 , which is a modified clinical isolate.

25. The virus of claim 24 wherein the clinical isolate is HSV1:

strain RH018A having the accession number ECACC 16121904;

strain RH004A having the accession number ECACC 16121902;

strain RH031A having the accession number ECACC 16121907;

strain RH040B having the accession number ECACC 16121908;

strain RH015A having the accession number ECACC 16121903;

strain RH021A having the accession number ECACC 16121905;

strain RH023A having the accession number ECACC 16121906; or

strain RH047A having the accession number ECACC 16121909.

26. The virus of claim 22 , wherein the at least three heterologous genes comprise:

(a) one or more immune stimulatory molecule-encoding genes; and/or

(b) one or more fusogenic protein-encoding genes.

27. The virus of claim 26 , wherein:

(a) the fusogenic protein is selected from the group consisting of vesicular stomatitis virus (VSV) G-protein, syncitin-1, syncitin-2, simian virus 5 (SV5) F-protein, measles virus (MV) H-protein, MV F-protein, respiratory syncytial virus (RSV) F-protein, and a glycoprotein from gibbon ape leukemia virus (GALV), murine leukemia virus (MLV), Mason-Pfizer monkey virus (MPMV), or equine infectious anaemia virus (EIAV) from which the R peptide has been deleted; and/or

(b) the immune stimulatory molecule is GM-CSF, IL-2, IL-12, IL-15, IL-18, IL-21, IL-24,a type I interferon, interferon gamma, a type III interferon, TNF alpha, an antagonist of TGF beta, an immune checkpoint antagonist, or an agonist of an immune potentiating pathway including an agonist of CD40,ICOS, GITR, 4-1-BB, 0X40, or flt3.

28. The virus of claim 27 , wherein:

(a) the fusogenic protein is the glycoprotein from gibbon ape leukemia virus (GALV) and has the R transmembrane peptide mutated or removed (GALV-R-);

(b) the agonist of an immune potentiating pathway is CD40 ligand (CD40L), ICOS ligand, GITR ligand, 4-1-BB ligand, 0X40 ligand, or flt3 ligand; and/or

(c) the immune checkpoint antagonist is a CTLA-4 inhibitor.

29. The virus of claim 28 , wherein the at least three heterologous genes comprise GALV-R- and one or more of GM-CSF, a CTLA-4 inhibitor, an agonist of CD40, an agonist of GITR, an agonist of 0X40, and an agonist of ICOS.

30. The virus of claim 28 , wherein the CTLA- 4 inhibitor is an anti-CTLA4 antibody, the agonist of CD40 is CD40L, the agonist of CD40 is 41BBL, the agonist of GITR is GITRL, the agonist of ICOS is ICOSL, and/or the agonist of 0X40 is OX40L.

31. The virus of claim 30 , wherein the HSV is a modified HSV1 strain RH018A having the accession number ECACC 16121904.

32. The virus of claim 22 , wherein the virus:

(a) does not express functional ICP34.5;

(b) does not express functional ICP47; and/or

(c) expresses the US 11 gene as an immediate early gene.

33. A pharmaceutical composition comprising the virus of claim 22 and a pharmaceutically acceptable carrier or diluent.

34. A product of manufacture comprising the virus of claim 22 in a sterile vial, ampoule or syringe.

35. A method of treating cancer, which comprises administering a therapeutically effective amount of the virus of claim 22 to a patient in need thereof.

36. A method according to claim 35 , which further comprises administering a therapeutically effective amount of a further anti-cancer agent to a patient in need thereof.

37. A method according to claim 36 , wherein the further anti-cancer agent is selected from an antagonist of an immune co-inhibitory pathway, an agonist of an immune co-stimulatory pathway, radiation and/or chemotherapy, an agent that targets a specific genetic mutation which occurs in tumors, an agent intended to induce an immune response to one or more tumor antigen(s) or neoantigen(s), a cellular product of T cells or NK cells, an agent intended to stimulate the STING, cGAS, TLR or other innate immune response and/or inflammatory pathway, a second virus optionally an oncolytic virus, an inhibitor of the indoleamine 2,3-dioxygenase (IDO) pathway, and combinations thereof.

38. A method according to claim 37 , wherein the antagonist of an immune co-inhibitory pathway is a CTLA-4 inhibitor, a PD-1 inhibitor, a PD-L1 inhibitor, a LAG-3 inhibitor, a TIM- 3 inhibitor, a VISTA inhibitor, a CSFIR inhibitor, an IDO inhibitor, a CEACAM1 inhibitor, a KIR inhibitor, a SLAMF7 inhibitor or a CD47 inhibitor, and/or the agonist of an immune co-stimulatory pathway is a GITR agonist, a 4-1-BB agonist, an 0X40 agonist, a CD40 agonist or an ICOS agonist.

39. A method according to claim 36 , wherein the further anti-cancer agent comprises an antibody.

40. A method according to claim 36 , wherein the virus and the further anti-cancer agent(s) are administered separately or concurrently.

41. A method according to claim 35 , wherein the cancer is a solid tumor.