Virus strains

• US 20030113348A1
• Filed: 11/12/2002
• Published: 06/19/2003
• Est. Priority Date: 01/21/2000
• Status: Active Grant

• Current Assignee: BioVex Ltd. (Amgen Inc.)
• Original Assignee: BioVex Ltd. (Amgen Inc.)
• Inventors: Coffin, Robert Stuart
• Granted Patent: US 7,063,835 B2
• Time in Patent Office: Days
• US Class Current: 424/230.1
• CPC Class Codes: A61K 2039/55516 Proteins; Peptides
• CPC Class Codes: A61K 2039/55522 Cytokines; Lymphokines; Interferons
• CPC Class Codes: A61K 35/763 Herpes virus
• 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 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/02 for peripheral neuropathies
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/04 specific for metastasis
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 15/869 Herpesviral vectors
• CPC Class Codes: C12N 2710/16021 Viruses as such, e.g. new isolates, mutants or their genomic sequences
• CPC Class Codes: C12N 2710/16033 Use of viral protein as therapeutic agent other than vaccine, e.g. apoptosis inducing or anti-inflammatory
• CPC Class Codes: C12N 2710/16632 Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
• CPC Class Codes: C12N 2710/16643 viral genome or elements thereof as genetic vector
• 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

Abstract

The present invention relates to non-laboratory virus strains, for example of herpes viruses such as HSV, with improved oncolytic and/or gene delivery capabilities as compared to laboratory virus strains.

Claims

1. Use of a modified, oncolytic, non-laboratory virus strain in the manufacture of a medicament for the oncolytic treatment of cancer.

2. Use according to claim 1 wherein said non-laboratory strain:

(a) has undergone one year or less in culture since isolation of its unmodified precursor strain from its host, or

(b) has undergone 100 or less cycles of serial passage since isolation of its unmodified precursor strain from its host, or

(c) has a greater ability than a reference laboratory strain with equivalent modifications to infect or replicate in a tumour cell, to kill tumour cells, or to spread between cells in tissue, or

(d) has substantially the ability of its unmodified precursor strain in respect of one or more of the properties defined in (c).

3. Use according to claim 2 wherein, in (c), said greater ability is a statistically significantly greater ability;

or wherein, in (d), substantially the ability is the same ability or an ability not statistically significantly different.

4. Use according to any one of the preceding claims wherein said non-laboratory virus strain is a strain of a herpes virus, adenovirus, picornavirus, retrovirus or alphavirus.

5. Use according to claim 4 wherein said non-laboratory virus strain is a strain of a herpes virus.

6. Use according to claim 5 wherein said non-laboratory virus strain is a strain of a herpes simplex virus (HSV).

7. Use according to claim 6 wherein HSV strain is a strain of HSV1 or HSV2.

8. Use according to any one of claims 5 to 7 wherein, in an HSV strain, the virus is modified such that it lacks one or more of a functional ICP34.5-encoding gene, a functional ICP6-encoding gene, a functional glycoprotein H-encoding gene, a functional thymidine kinase-encoding gene;

or, in a non-HSV strain, the virus lacks a functional gene equivalent to one of said HSV genes.

9. Use according to any one of claims 5 to 8 wherein the strain is an HSV strain and the virus lacks a functional ICP34.5-encoding gene.

10. Use according to claim 9 wherein the virus further lacks a functional ICP47 gene.

11. Use according to any one of the preceding claims wherein said non-laboratory virus strain further comprises a heterologous gene.

12. Use according to claim 11 wherein said heterologous gene is a gene capable of modifying immune responses.

13. Use according to claim 12 wherein said heterologous gene capable of modifying immune responses encodes an immune stimulatory polypeptide or another gene product capable of modifying immune responses, a prodrug activator, a tumour suppressor or a pro-apoptotic gene product.

14. Use according to claim 13 wherein said immune stimulatory polypeptide is granulocyte macrophage colony-stimulating factor (GMCSF), another cytokine or chemokine, RANTES, B7.1 or B7.2 or IL12, or wherein the prodrug activator is nitroreductase or cytochrome p450, or wherein the tumour suppressor is p53.

15. Use according to any one claims 2 to 14 wherein the non-laboratory strain is an HSV strain and the reference strain as defined in claim 2 is HSV1 strain 17+, HSV1 strain F or HSV1 strain KOS with equivalent modifications to the non-laboratory strain.

30. A method of determining the suitability of a non-laboratory virus strain for modification into a modified strain as defined in any one of the preceding claims, comprising:

(i) optionally, isolating a non-laboratory virus strain from a host;

(ii) providing said non-laboratory virus strain;

(iii) assessing the properties of the virus in respect of one or more of the characteristics defined in part (c) or (d) of claim 2 or part (c), (d) or (e) of claim 17;

and, optionally,

(iv) selecting for modification virus strains with desirable properties.

31. A method of determining the suitability of a non-laboratory virus strain for modification into a modified oncolytic strain as defined in any one of claims 1 to 15, comprising:

(i) optionally, isolating a non-laboratory virus strain from a host;

(ii) assessing the growth of the virus in one or more types of tumour cell; and

optionally

(iii) selecting for modification virus strains with a high growth rate.

32. A method according to claim 31 wherein the virus is as defined in any one of claims 4 to 7.

33. A method of determining the suitability of a non-laboratory virus strain for modification into a modified oncolytic strain, comprising:

(i) providing a non-laboratory virus strain, optionally one selected by a method according to step (ii) of claim 31 or 32;

(ii) modifying said strain such that it becomes oncolytic; and

(iii) assessing the ability of said modified, oncolytic non-laboratory strain to kill tumour cells; and

optionally

(iv) selecting strains that exhibit high tumour cell-killing capacity for further modifications; and

optionally

(v) carrying out further modifications.

34. A method according to claim 33 wherein said virus is as defined in any one of claims 4 to 7.

35. A method according to claim 33 wherein the virus is a herpes virus and, in an HSV strain, it is modified by rendering non-functional one or more genes selected from the genes encoding ICP34.5, ICP6 and thymidine kinase;

or, in a non-HSV strain, it is modified by rendering non-functional a gene or genes equivalent to said HSV gene or genes.

36. A method according to any one of claims 33 to 35 wherein the tumour cell-killing capacity of said modified, oncolytic non-laboratory strain is measured by comparison to the tumour cell-killing capacity of a reference laboratory strain with the same gene or genes rendered non-functional.

37. A method according to claim 36 wherein the non-laboratory strain is an HSV strain and the reference stain is HSV1 strain 17+, HSV strain F or HSV strain KOS.

38. A method of determining whether a gene enhances the anti-tumour effects of a virus comprising:

(i) providing a modified, oncolytic non-laboratory strain as defined in any one of claims 1 to 15;

(ii) inserting said gene into said virus as a heterologous gene; and

(iii) assessing the ability of said modified, oncolytic non-laboratory strain to kill tumour cells compared to the ability of the precursor strain provided in step (i).

39. A method of producing a modified, oncolytic, non-laboratory virus strain comprising:

(i) isolating from a host a non-laboratory strain of a virus;

(ii) optionally determining its suitability for modification as set out in claim 33;

and

(iii) modifying it to render it oncolytic, and optionally

(iv) carrying out further modifications.

40. A method according to claim 39 wherein the resulting modified, oncolytic, non-laboratory virus is as defined in any one of claims 2 to 15.

43. A modified, oncolytic, non-laboratory virus strain as defined in any one of claims 1 to 15.

45. A modified non-laboratory virus strain according to claim 43 or 44 which is a herpes virus strain.

46. A modified non-laboratory virus strain according to claim 45 which is a herpes simplex virus (HSV) strain.

50. A non-laboratory virus strain identified by, or produced in the course of, a method according to any one of claims 30 to 42.

51. Use of a gene identified as enhancing the anti-tumour effects of a virus by a method according to claim 38, or of a gene product encoded by said gene, in the manufacture of a medicament for the treatment or prevention of cancer.

52. A modified non-laboratory virus strain obtained or obtainable by a method according to any one of claims 39 to 42.

54. An HSV1 strain derived from JS1 as defined in claim 53 and having the characteristics set out in any one of claims 2 to 29.

55. An HSV1 strain derived from JS1 as defined in claim 54 and modified such that it lacks a functional ICP34.5 gene, and optionally a functional ICP47 gene;

and optionally further comprises a heterologous gene encoding GMCSF.

58. A method of treating a tumour in an individual in need thereof by administering to said individual an effective amount of a virus as defined in claim 1.

16. Use of a modified, replication incompetent, non-laboratory virus strain comprising a heterologous gene in the manufacture of a medicament for the delivery of said gene to a subject.

17. Use according to claim 16 wherein said non-laboratory virus strain:

(a) has undergone one year or less in culture since isolation of its unmodified precursor strain from its host, or

(b) has undergone 100 or less cycles of serial passage since isolation of its unmodified precursor strain from its host, or

(c) has a greater ability than a reference laboratory strain with equivalent modifications to infect a target cell, or

(d) has a greater ability than a reference laboratory strain with equivalent modifications, to infect a neuron, to spread between cells in nervous tissue, to be transported within an axon, to infect a dendritic cell or to induce an immune response, or

(e) has substantially the ability of its unmodified precursor strain in respect of one or more of the properties defined in (c) or (d).

18. Use according to claim 17 wherein, in (c), a greater ability is a statistically significantly greater ability;

or, in (d), substantially the ability is the same ability or an ability not statistically significantly different.

19. Use according to claim 18 wherein the non-laboratory virus strain is as defined in any one of claims 4 to 7.

20. Use according to claim 19 wherein in an HSV strain, the virus is modified such that it lacks one, two, three or all of a functional ICP27-encoding gene, a functional ICP4-encoding gene, a functional ICP0-encoding gene, or a functional ICP22-encoding gene;

or, in an non-HSV strain, the virus lacks a functional gene equivalent to one of said HSV genes; and

/or, in an HSV strain, the virus lacks a functional vmw65 gene due to a mutation in said gene which abolishes its transcriptional-activation activity;

or in a non-HSV strain, the virus lacks a functional gene equivalent to vmw65 due to a mutation in said gene which abolishes its transcriptional-activation activity.

21. Use according to any one of claims 16 to 20 wherein the medicament is for use in treating or preventing a peripheral nervous system disorder by administering said medicament to a peripheral nerve and said heterologous gene encodes a polypeptide or antisense RNA of therapeutic use in the treatment of said peripheral nervous system disorder.

22. A method of determining whether a gene has an effect on a phenotype associated with a peripheral nervous system disorder or on a cell of the peripheral nervous system which is relevant to a peripheral nervous system disorder, which method comprises:

(i) inoculating a replication incompetent herpes virus as defined in any one of claims 16 to 21 comprising a heterologous gene into a peripheral nerve; and

(ii) monitoring a phenotype of said disorder or an effect of expression of said gene on said cell to determine thereby whether said gene has an effect relevant to said disorder.

23. Use according to any one of claims 16 to 20 wherein the medicament is for use in treating or preventing a central nervous system disorder, the heterologous gene encodes a polypeptide or antisense RNA of therapeutic use in treating said disorder and the replication incompetent herpes virus comprises:

(a) one or more mutations which prevent or reduce the expression of at least two immediate early genes; and

(b) a heterologous gene operably linked to a promoter active during herpes virus latency.

24. Use according to claim 27 wherein said strain is an HSV strain and said immediate early gene is, or genes are, one, two, three, or all of the genes encoding ICP0, ICP4, ICP22 and ICP27.

25. A method of determining whether a gene has an effect on a phenotype associated with a central nervous system disorder or on a cell of the central nervous system which is relevant to a central nervous system disorder, which method comprises:

(i) inoculating into a cell of the central nervous system with a replication incompetent herpes virus as defined in claim 23 or 24; and

(ii) monitoring a phenotype of said disorder or an effect of expression of said gene on said cell to determine thereby whether said gene has an effect on said cell or said phenotype.

26. Use according to any one of claims 16 to 20 wherein:

the herpes virus is an attenuated herpes virus capable of efficiently infecting a dendritic cell without preventing the infected cell from stimulating an immune response;

the medicament is for use in a method of treating or preventing a pathogenic infection or cancer;

which method comprises infecting dendritic cells with the said virus; and

the heterologous gene encodes an antigen associated with said infection or cancer.

27. Use according to claim 26 wherein, in an HSV strain, said virus lacks a functional UL43 gene, and/or a functional vhs gene, or, in a non-HSV strain, lacks a functional equivalent of UL43 and/or vhs;

and optionally, in an HSV strain, further lacks a functional ICP34.5 gene, or, in a non-HSV strain, a functional equivalent of ICP34.5; and

optionally, in an HSV strain, lacks a functional vmw65 gene due to a mutation in said gene which abolishes its transcriptional-activation activity, or, in a non-HSV strain, lacks a functional gene equivalent to vmw65 due to a mutation in said gene which abolishes its transcriptional-activation activity; and

optionally lacks at least one functional immediate early gene.

28. Use according to claim 27 wherein said strain is an HSV strain and said immediate early gene is, or genes are, one, two, three or all of the genes encoding ICP0, ICP4, ICP22 and ICP27.

29. A method of determining whether a gene encodes an antigen associated with a pathogenic infection or cancer, which method comprises infecting a dendritic cell or a macrophage with a virus as defined in any one of claims 26 to 28 and monitoring antigen presentation of the polypeptide product of said gene, or an effect of expression of said gene, or a phenotype of said pathogenic infection or cancer to determine thereby whether said gene encodes an antigen associated with said infection or cancer and which itself has therapeutic potential or is a target for therapeutic intervention.

42. A method according to claim 41 wherein the resulting modified non-laboratory virus strain is as defined in any one of claims 16 to 29.

44. A modified non-laboratory virus strain comprising a heterologous gene, as defined in any one of claims 16 to 29.

47. Use of a gene identified as having an effect on a phenotype associated with a peripheral nervous system disorder or on a cell of the peripheral nervous system which is relevant to a peripheral nervous system disorder by a method according to claim 22, or of a gene product encoded by said gene in the manufacture of a medicament for the treatment of a peripheral nervous system disorder.

48. Use of a gene identified as having an effect on a phenotype associated with a central nervous system disorder or on a cell of the central nervous system which is relevant to a central nervous system disorder by a method according to claim 23, or of a gene product encoded by said gene in the manufacture of a medicament for the treatment of a central nervous system disorder.

49. Use of a gene identified as encoding an antigen associated with a pathogenic infection or cancer by a method according to claim 29, or of an antigen encoded by said gene in the manufacture of a medicament for the treatment or prevention of said infection or cancer.

59. A method of delivering a gene to an individual in need thereof by administering to said individual an effective amount of a virus as defined in claim 16.

60. A method of treating or preventing a peripheral nervous system disorder by administering to a peripheral nerve of an individual in need thereof an effective amount of a virus as defined in claim 21.

61. A method of treating or preventing a central nervous system disorder comprising administering to an individual in need thereof a virus as defined in claim 23.

41. A method of producing a modified non-laboratory virus strain comprising:

(i) providing a non-laboratory strain of a virus;

(ii) modifying it to render it replication incompetent and,

(iii) inserting a heterologous gene.

53. HSV1 strain JS1 as deposited at the European Collection of Cell Cultures (ECACC) under provisional accession number 01010209, or an HSV1 strain derived therefrom.

56. A pharmaceutical composition comprising a virus as defined in any one of claims 53 to 55.

57. A virus as defined in any one of claims 53 to 55 for use in the treatment of the human or animal body.