Oncolytic virus for cancer therapy
It is an object of the present invention to develop a virus preparation comprising an oncolytic HSV having, in vivo, more effective cancer cell-killing activity than the antitumor effects of existing oncolytic HSVs. Specifically, the present invention relates to a virus preparation for the treatment of a cancer, comprising an HSV (herpes simplex virus) having a receptor-retargeted gD mutation and at least one membrane fusion activity-promoting region on the genome, and a method for treating cancer using the aforementioned virus preparation.
1. A virus preparation for the treatment of a cancer, comprising an HSV (herpes simplex virus) having a receptor-retargeted gD mutation and at least one membrane fusion activity-promoting region on the genome,
wherein the membrane fusion activity-promoting region is a region having a syn mutation, and
wherein the syn mutation is a mutation comprising an amino acid mutation at position 858 of SEQ ID NO: 2 which is a substitution with cysteine or histidine.
2. The virus preparation according to claim 1 , wherein the receptor-retargeted gD mutation is a mutation to delete binding ability to nectin-1, the mutation to delete binding ability to nectin-1 being a deletion of all of the amino acids at positions 6 to 38 of SEQ ID NO: 1, a deletion of all of the amino acids at positions 61 to 218 of SEQ ID NO: 1, a mutation of the amino acids at positions 3 and 38 of SEQ ID NO: 1, and/or a mutation of the amino acids at positions 222 and 223 of SEQ ID NO: 1.
3. The virus preparation according to claim 2 , wherein the amino acid mutation at position 3 of SEQ ID NO: 1 is a deletion or a substitution with cysteine, the amino acid mutation at position 38 of SEQ ID NO: 1 is a substitution with cysteine, the amino acid mutation at position 222 of SEQ ID NO: 1 is a substitution with asparagine, and the amino acid mutation at position 223 of SEQ ID NO: 1 is a substitution with isoleucine.
4. The virus preparation according to claim 1 , wherein the receptor-retargeted gD mutation is a mutation to delete binding ability to HVEM and 3-OS-HS, the mutation to delete binding ability to HVEM and 3-OS-HS being a deletion of all or a part of the amino acids at positions 2 to 38 of SEQ ID NO: 1, a deletion of all of the amino acids at positions 61 to 218 of SEQ ID NO: 1, an amino acid mutation at position 27 of SEQ ID NO: 1, an amino acid mutation at position 29 of SEQ ID NO: 1, and/or an amino acid mutation at position 30 of SEQ ID NO: 1.
5. The virus preparation according to claim 4 , wherein: the deletion of a part of the amino acids at positions 2 to 38 of SEQ ID NO: 1 is any of a deletion of the amino acids at positions 2 to 24, a deletion of the amino acids at positions 7 to 11, a deletion of the amino acids at positions 7 to 32, or a deletion of the amino acids at positions 6 to 38; the amino acid mutation at position 27 is a substitution with alanine, proline or arginine; the amino acid mutation at position 29 is a substitution with alanine; and the amino acid mutation at position 30 is a substitution with alanine.
6. The virus preparation according to claim 1 , wherein the syn mutation further comprises a mutation described in the following (a), (b), (c) and/or (d):
(a) an amino acid mutation at position 796, an amino acid mutation at position 800, an amino acid mutation at position 813, an amino acid mutation at position 817, an amino acid mutation at position 854, an amino acid mutation at position 855, an insertion of an amino acid between the amino acids at positions 816 and 817, a nonsense mutation in the amino acid at position 877, and/or a nonsense mutation in the amino acid at position 869, of SEQ ID NO: 2;
(b) an amino acid mutation at position 33, an amino acid mutation at position 40, an amino acid mutation at position 86, an amino acid mutation at position 99, an amino acid mutation at position 111, an amino acid mutation at position 121, an amino acid mutation at position 243, an amino acid mutation at position 304, and/or an amino acid mutation at position 310, of SEQ ID NO: 3;
(c) an amino acid mutation at position 49, amino acid mutations at positions 49, 50 and 51, an amino acid mutation at position 209, amino acid mutations at positions 209, 212 and 213, a nonsense mutation in the amino acid at position 217, of SEQ ID NO: 4, and/or a deletion of all of the amino acids shown in SEQ ID NO: 4; and/or
(d) amino acid mutations at position 62, 63 and 64, of SEQ ID NO: 5.
7. The virus preparation according to claim 6 , wherein:
the amino acid mutation at position 796 of SEQ ID NO: 2 is a substitution with cysteine, the amino acid mutation at position 800 of SEQ ID NO: 2 is a substitution with tryptophan, the amino acid mutation at position 813 of SEQ ID NO: 2 is a substitution with isoleucine, the amino acid mutation at position 817 of SEQ ID NO: 2 is a substitution with histidine or proline, the amino acid mutation at position 854 of SEQ ID NO: 2 is a substitution with phenylalanine, and the amino acid mutation at position 855 of SEQ ID NO: 2 is a substitution with valine;
the amino acid mutation at position 33 of SEQ ID NO: 3 is a substitution with serine, the amino acid mutation at position 40 of SEQ ID NO: 3 is a substitution with valine or threonine, the amino acid mutation at position 86 of SEQ ID NO: 3 is a substitution with proline, the amino acid mutation at position 99 of SEQ ID NO: 3 is a substitution with asparagine, the amino acid mutation at position 111 of SEQ ID NO: 3 is a substitution with valine, the amino acid mutation at position 121 of SEQ ID NO: 3 is a substitution with isoleucine, the amino acid mutation at position 243 of SEQ ID NO: 3 is a substitution with tyrosine, the amino acid mutation at position 304 of SEQ ID NO: 3 is a substitution with proline, and the amino acid mutation at position 310 of SEQ ID NO: 3 is a substitution with leucine;
the amino acid mutation at position 49 of SEQ ID NO: 4 is a substitution with alanine, the amino acid mutation at position 50 of SEQ ID NO: 4 is a substitution with alanine, the amino acid mutation at position 51 of SEQ ID NO: 4 is a substitution with alanine, the amino acid mutation at position 209 of SEQ ID NO: 4 is a substitution with alanine, the amino acid mutation at position 212 of SEQ ID NO: 4 is a substitution with alanine, and the amino acid mutation at position 213 of SEQ ID NO: 4 is a substitution with alanine; and
the amino acid mutation at position 62 of SEQ ID NO: 5 is a substitution with glycine, the amino acid mutation at position 63 of SEQ ID NO: 5 is a substitution with valine, and the amino acid mutation at position 64 of SEQ ID NO: 5 is a substitution with serine.
8. The virus preparation according to claim 1 , wherein a reporter gene and/or a therapeutic gene are incorporated into the genome of the HSV.
9. A therapeutic method for treating a cancer, comprising administering a therapeutically-effective amount of the virus preparation according to claim 1 to a subject in need thereof.
10. A therapeutic method for treating a cancer, comprising administering a therapeutically-effective amount of the virus preparation according to claim 2 to a subject in need thereof.
11. A therapeutic method for treating a cancer, comprising administering a therapeutically-effective amount of the virus preparation according to claim 3 to a subject in need thereof.
12. A therapeutic method for treating a cancer, comprising administering a therapeutically-effective amount of the virus preparation according to claim 4 to a subject in need thereof.
13. A therapeutic method for treating a cancer, comprising administering a therapeutically-effective amount of the virus preparation according to claim 5 to a subject in need thereof.
14. A therapeutic method for treating a cancer, comprising administering a therapeutically-effective amount of the virus preparation according to claim 6 to a subject in need thereof.
15. A therapeutic method for treating a cancer, comprising administering a therapeutically-effective amount of the virus preparation according to claim 7 to a subject in need thereof.
16. A therapeutic method for treating a cancer, comprising administering a therapeutically-effective amount of the virus preparation according to claim 8 to a subject in need thereof.
17. The virus preparation according to claim 1 , wherein the receptor-retargeted gD mutation is a deletion of the amino acids at positions 2 to 24 of SEQ ID NO:1 and an amino acid mutation at position 38 of SEQ ID NO:1 which is a substitution with cysteine, wherein the syn mutation further comprises an amino acid mutation at position 40 of SEQ ID NO: 3 which is a substitution with valine or threonine.
18. A therapeutic method for treating a cancer, comprising administering a therapeutically-effective amount of the virus preparation according to claim 17 to a subject in need thereof.