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Oncolytic virus improved in safety and anticancer effect

A technology of oncolytic virus and recombinant virus vector, which is applied in the direction of resisting vector-borne diseases, viruses, and viral peptides, and can solve the controversial problems of other cytotoxic effects, so as to control adverse side effects, inhibit virus replication, and improve anti-cancer The effect of action

Pending Publication Date: 2020-08-18
拜耳诺克斯有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, other cytotoxic effects expected from co-administration of GCV remain controversial

Method used

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  • Oncolytic virus improved in safety and anticancer effect
  • Oncolytic virus improved in safety and anticancer effect
  • Oncolytic virus improved in safety and anticancer effect

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0132] Embodiment 1. Preparation of recombinant vaccinia virus (OTS-412)

Embodiment 11

[0133] Example 1.1. Construction of the shuttle plasmid vector

[0134] Wild-type vaccinia virus (NYC Department of Health strain, VR-1536) was purchased from the American Type Culture Collection (ATCC). For recombination, pUC57amp+ (Genewiz, USA) containing the HSV1-TK gene (pSE / L promoter) and firefly luciferase reporter gene (p7.5 promoter) was used as a shuttle plasmid vector ( figure 1 ).

Embodiment 12

[0135] Example 1.2. Preparation of recombinant vaccinia virus

[0136] To ensure the safety of the recombinant virus, 4 × 10 5 cells / well concentration, HeLa cells (ATCC) were prepared in EMEM medium containing 10% fetal bovine serum. Cells were then treated with 0.05 MOI of wild-type vaccinia virus. After 2 hours, replace the medium with EMEM medium containing 2% fetal bovine serum, and then TM Polymer (Clontech 631317, USA), cells were transfected with 4 μg of the linearized shuttle plasmid vector constructed in Example 1.1. After 4 hours of incubation, the medium was replaced with fresh EMEM medium containing 2% fetal bovine serum, and the cells were further incubated for 72 hours. The luciferase activity in HeLa cells was confirmed, and thus the recombinant vaccinia virus containing HSV1-TK gene was obtained.

[0137]Thereafter, from a TK-osteosarcoma cell line (osteosarcoma 143TK-) in the presence of BrdU (thymidine analogue, 15 μg / ml) under biochemical conditions for...

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Abstract

The present invention relates to an oncolytic virus improved in safety and anticancer effect and a use thereof. The oncolytic virus improved in safety and anticancer effect of the present invention isobtained by inserting an HSV-TK fragment-encoding gene into a TK gene region to delete TK of Vaccinia virus. In addition, the oncolytic virus of the present invention expresses an HSV-TK fragment tophosphorylate GCV so that cancer cells infected with the oncolytic virus and even their neighboring cancer cells can be killed. In addition, GCV is also involved in the suppression of viral proliferation and thus can control side effects caused by a virus even upon the administration of a high dose of the virus. Furthermore, an anticancer effect is increased even though the number of viral particles is reduced due to the suppression of GCV against virus proliferation. Therefore, the oncolytic virus improved in safety and anticancer effect of the present invention can be effectively used for the treatment of cancer.

Description

technical field [0001] The present invention relates to an oncolytic virus with improved safety and anticancer effect and its use. Background technique [0002] With the full application of gene recombination technology, clinical studies using oncolytic viruses with improved tumor selectivity and anticancer efficacy have been initiated. The first recombinant oncolytic virus reported in the literature was the herpes simplex virus. Since then, the use of other viruses for oncolysis has been actively studied (Martuza et al., 1991; Hwang et al., 2011; Kaufaman et al., 2015; Khuri et al., 2000; Park et al., 2008). [0003] The use of oncolytic viruses has received considerable attention due to the recent successful commercialization of herpes virus-based T-Vec (Talimogene laherparepvec) in the treatment of advanced melanoma in the United States and Europe. Meanwhile, vaccinia viruses lacking the thymidine kinase (TK) gene have great clinical utility, but there are limitations i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/86A61K31/519A61K35/768A61P35/00C12N9/12
CPCA61K35/768A61P35/00C12N15/86Y02A50/30A61K45/06A61K31/522C12N9/1211C12N2710/24143C12N2710/24132C07K14/005C12N2710/16622A61K2300/00A61K31/519C12Y207/01021C12N2710/24141A61K9/0019A61K31/708C12N2710/24111
Inventor 黄泰皓赵懞
Owner 拜耳诺克斯有限公司
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