Recombinant herpes simplex virus and use thereof

Patent metadata

Field	Value
Publication number	US-11680248-B2
Application number	US-201816491699-A
Country	US
Kind code	B2
Filing date	Feb 28, 2018
Priority date	Mar 9, 2017
Publication date	Jun 20, 2023
Grant date	Jun 20, 2023

Abstract

The invention relates to the field of virology and tumor therapy. In particular, the present invention provides a recombinant herpes simplex virus (HSV) capable of specifically replicating at a high level in a tumor cell and effectively killing the tumor cell, but replicating at low levels in normal cells, thereby the recombinant herpes simplex virus of the present invention not only has high lethality against tumor cells, but also has significantly decreased side effects (especially neurotoxicity). Further, the present invention relates to a viral vector constructed based on the recombinant herpes simplex virus, a pharmaceutical composition comprising the recombinant herpes simplex virus or the viral vector, and the use of the recombinant herpes simplex virus or the viral vector. The recombinant herpes simplex virus of the present invention can be used to infect and kill tumor cells, and can be used for gene drug delivery into tumor cells for gene therapy.

First claim

What is claimed is:

1. A recombinant herpes simplex virus (HSV), which does not express a functional ICP0 protein and ICP34.5 protein; but is capable of expressing a functional UL43 protein, a functional UL41 protein, a functional UL48 protein, or any combination thereof; and wherein the genome of the recombinant HSV comprises the following modifications:
two copies of the ICP0 gene each independently comprising a loss-of-function mutation or which is deleted or substituted with an exogenous nucleotide sequence;
two copies of the ICP34.5 gene each independently comprising a loss-of-function mutation or which is deleted or substituted with an exogenous nucleotide sequence; and
wherein the genome of the recombinant HSV further comprises a modification in which a native promoter of one or more HSV genes is substituted with a tumor-specific promoter.

2. The recombinant HSV according to claim 1 , wherein the genome of the recombinant HSV comprises:
(1) a UL43 gene capable of expressing a functional UL43 protein,
(2) a UL41 gene capable of expressing a functional UL41 protein,
(3) a UL48 gene capable of expressing a functional UL48 protein, or
(4) any combination of (1) to (3).

3. The recombinant HSV according to claim 1 , wherein the recombinant HSV further comprises an additional exogenous nucleotide sequence.

4. A viral vector, comprising the genome of the recombinant HSV according to claim 1 .

5. An isolated host cell, which comprises one of the following:
(1) the recombinant HSV according to claim 1 , or
(2) the genome of the recombinant HSV of claim 1 , or
(3) a viral vector comprising the genome of the recombinant HSV of claim 1 .

6. A method of obtaining the recombinant HSV according to claim 1 , comprising:
(1) cultivating a host cell, which is infected with the recombinant HSV, or comprises a genome of the recombinant HSV, or is transfected with a viral vector comprising a genome of the recombinant HSV;
(2) collecting and lysing the host cell after the host cell shows signs of infection, to obtain a lysate of the host cell; and
(3) recovering the recombinant HSV from the lysate.

7. A pharmaceutical composition, which comprises one or more of the following and a pharmaceutically acceptable carrier or excipient:
(1) the recombinant HSV according to claim 1 ,
(2) the genome of the recombinant HSV of claim 1 , and
(3) a viral vector comprising the genome of the recombinant HSV of claim 1 .

8. A method of treating a tumor, which comprises administering to a subject in need thereof a therapeutically effective amount of one or more of the following:
(1) the recombinant HSV according to claim 1 ,
(2) a viral vector comprising the genome of the recombinant HSV of claims 1 , and
(3) a pharmaceutical composition comprising the recombinant HSV of (1) or the viral vector of (2).

9. The recombinant HSV according to claim 1 , wherein in the genome of the recombinant HSV:
(1) the two copies of the ICP0 genes each independently comprises a loss-of-function mutation; and, the two copies of the ICP34.5 gene each independently comprises a loss-of-function mutation; or
(2) the two copies of the ICP0 genes each independently comprises a loss-of-function mutation; and the two copies of the ICP34.5 gene are deleted; or
(3) the two copies of the ICP0 genes each independently comprises a loss-of-function mutation; and the two copies of the ICP34.5 gene each is independently substituted with an exogenous nucleotide sequence; or
(4) the two copies of the ICP0 gene are deleted; and, the two copies of the ICP34.5 gene each independently comprises a loss-of-function mutation; or
(5) the two copies of the ICP0 gene are deleted; and the two copies of the ICP34.5 gene are deleted; or
(6) the two copies of the ICP0 gene are deleted; and the two copies of the ICP34.5 gene each is independently substituted with an exogenous nucleotide sequence; or
(7) the two copies of the ICP0 gene each is independently substituted with an exogenous nucleotide sequence; and, the two copies of the ICP34.5 gene each independently comprises a loss-of-function mutation; or
(8) the two copies of the ICP0 gene each is independently substituted with an exogenous nucleotide sequence; and the two copies of ICP34.5 genes are deleted; or
(9) the two copies of the ICP0 gene each is independently substituted with an exogenous nucleotide sequence; and the two copies of the ICP34.5 gene each is independently substituted with an exogenous nucleotide sequence.

10. The recombinant HSV according to claim 9 , wherein
a) the loss-of-function mutation each is independently selected from the group consisting of: missense mutation, nonsense mutation, frameshift mutation, base deletion, base substitution, base addition, and any combination thereof;
b) the exogenous nucleotide sequence each independently encodes a foreign protein selected from the group consisting of: fluorescent protein, immunomodulatory polypeptide, cytokine, chemokine, antibody, and cytotoxic peptide, or
c) a combination of a) and b).

11. The recombinant HSV according to claim 10 , having any one or more of the following:
(1) the fluorescent protein is selected from the group consisting of green fluorescent protein, red fluorescent protein, blue fluorescent protein, yellow fluorescent protein, and any combination thereof;
(2) the immunomodulatory polypeptide is selected from the group consisting of CD40L, OX40L, inducible costimulatory molecule (ICOS), FTL3L, LIGHT, CD137L, CD70, 4-1BB, GITR, CD28, and any combination thereof;
(3) the cytokine is selected from the group consisting of interleukin, interferon, tumor necrosis factor, colony stimulating factor, and any combination thereof;
(4) the chemokine is selected from the group consisting of CCL2, RANTES, CCL7, CCL9, CCL10, CCL12, CCL15, CCL19, CCL21, CCL20, XCL-1, and any combination thereof;
(5) the cytotoxic peptide is selected from the group consisting of thymidine kinase TK (TK/GCV), TRAIL, FasL, and any combination thereof; and
(6) the antibody is selected from the group consisting of anti-PD-1 antibody, anti-PD-L1 antibody, anti-TIGIT antibody, anti-BTLA antibody, anti-CTLA-4 antibody, anti-Tim-3 antibody, anti-Lag-3 antibody, anti-CD137 antibody, anti-OX40 antibody, anti-GITR antibody, anti-CD73 antibody, anti-KIR antibody, anti-ICOS antibody, anti-CSF1R antibody, anti-EGFR antibody, anti-VEGFR antibody, anti-HER2 antibody, anti-PDGFR antibody, and any combination thereof.

12. The recombinant HSV according to claim 2 , wherein the recombinant HSV is further characterized by:
(1) one or more non-essential genes are deleted or mutated;
(2) except for the two copies of the ICP0 gene and the two copies of the ICP34.5 gene, the genome of the recombinant HSV comprises all other genes of the wild-type HSV, and none of the other genes comprises a loss-of-function mutation; or
(3) the tumor-specific promoter is a promoter of hTERT.

13. The recombinant HSV according to claim 12 , characterized by any one or more of the following:
(1) the non-essential gene is selected from the group consisting of UL3 gene, UL4 gene, UL14 gene, UL16 gene, UL21 gene, UL24 gene, UL31 gene, UL32 gene, US3 gene, UL51 gene, UL55 gene, UL56 gene, US2 gene, US12 gene, LAT gene, nucleotide fragment corresponding to SEQ ID NO: 23, and any combination thereof;
(2) the non-essential gene comprises a loss-of-function mutation, or is substituted with an exogenous nucleotide sequence; and
(3) the tumor-specific promoter is a promoter of hTERT having the sequence set forth in SEQ ID NO: 5.

14. The recombinant HSV according to claim 2 , wherein the genom

Assignees

• Univ Xiamen
• Yang Sheng Tang Co Ltd

Inventors

• Huang Chenghao
• Luo Yong
• Yuan Quan
• Zhang Jun
• Xia Ningshao

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• C12N2710/16643

  viral genome or elements thereof as genetic vector · CPC title

• C12N15/86

  Viral vectors · CPC title

• C12N2710/16622

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title

• A61K35/763 Primary

  Herpes virus · CPC title