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Genetically engineered oncolytic vaccinia viruses and methods of use thereof

A vaccinia virus and oncolytic technology, applied in genetic engineering, plant genetic improvement, virus, etc., can solve problems such as inability to exert therapeutic effects

Pending Publication Date: 2022-06-03
ASTELLAS PHARMA INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, oncolytic vaccinia viruses expressing an immunostimulatory molecule may be rapidly cleared by a strong immune response stimulated by the molecule, rendering them unable to exert therapeutic effects

Method used

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  • Genetically engineered oncolytic vaccinia viruses and methods of use thereof
  • Genetically engineered oncolytic vaccinia viruses and methods of use thereof
  • Genetically engineered oncolytic vaccinia viruses and methods of use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0339] Example 1. Cytotoxicity of vaccinia virus carrying hIL12 and hIL7 on human tumor cells

[0340] This study was conducted to determine whether vaccinia virus carrying hIL12 and hIL7 showed cytotoxic effects in the following human cancer cell lines: human colorectal cancer (COLO 741) cells, human glioblastoma (U-87MG) cells and human cholangiocarcinoma (HuCCT1) cells.

[0341] All cells were infected with vaccinia virus carrying hIL12 and hIL7 at different multiplicities of infection (MOI) (0, 0.1, 1, 10 and 100). 45 days after infection, with CellTiter- The 2.0 assay measures cell viability. Cell viability was calculated by setting uninfected cells (MOI 0) and medium control wells without cells to 100% and 0% viability, respectively. One experiment was performed and data are presented as the mean of three replicate measurements.

[0342] At 4 days post-infection, cell viability dropped to <10% (Table 1). These results indicate that vaccinia viruses carrying hIL12 a...

Embodiment 2

[0346] Example 2. Cytotoxic activity of vaccinia virus carrying hIL12 and hIL7 against various human cancer cell lines

[0347] This study was conducted to further examine whether hIL12 and hIL7-carrying vaccinia viruses exhibited cytotoxic effects on 24 human cancer cell lines.

[0348] Cells were infected with vaccinia virus carrying hIL12 and hIL7 at different multiplicities of infection (MOI). 5 days after infection, with CellTiter- The luminescent cell viability assay measures cell viability. Cell viability was calculated by setting uninfected cells (MOI 0) and medium control wells without cells to 100% and 0% viability, respectively. One experiment was performed and data are presented as the mean of three replicate measurements.

[0349] like figure 1 As shown, vaccinia viruses carrying hIL12 and hIL7 were cytotoxic to all human cancer cells examined 5 days after infection at MOIs of 1.0, 10 or 100.

Embodiment 3

[0350] Example 3. Replication of vaccinia virus carrying hIL12 and hIL7 in human cancer cells or normal cells

[0351] This study was conducted to examine whether hIL12 and hIL7-carrying vaccinia viruses replicate selectively in human cancer cells relative to normal cells.

[0352] Human cancer cells (NCI-H520, HARA, LK-2 and LUDLU 1) or normal human bronchial epithelial cells (HBEpC) were infected with vaccinia virus carrying hIL12 and hIL7 at an MOI of 1, or with vehicle (MOI 0). Cells were harvested at 6 h or 24 h post infection and the DNA amount of vaccinia virus carrying hIL12 and hIL7 was measured by standard quantitative polymerase chain reaction (qPCR) with primers designed to amplify the vaccinia virus hemagglutinin (HA) J7R Gene. Values ​​are normalized to the 18s ribosomal RNA gene and presented as the mean of two replicate measurements.

[0353] like figure 2 shown that the amount of genomic DNA of hIL12 and hIL7-carrying vaccinia virus was detected in all hum...

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Abstract

The present invention provides pharmaceutical compositions comprising an oncolytic vaccinia virus and methods of using such pharmaceutical compositions to treat subjects having cancer.

Description

[0001] Related applications [0002] This application claims the benefit of or priority to US Provisional Application No. 62 / 893,316, filed August 29, 2019, the entire contents of which are incorporated herein by reference. [0003] This application is related to US Patent Publication No. 2017 / 0340687, Japanese Patent Application Nos. JP 2018 223349 and JP 2018 179632, the entire contents of each of which are incorporated herein by reference. [0004] sequence listing [0005] This application contains a Sequence Listing electronically filed in ASCII format and is incorporated herein by reference in its entirety. Said ASCII copy was created on August 25, 2020, named 127206_03920_SL.txt, and is 4,095 bytes in size. Background technique [0006] Various techniques have been developed recently to use viruses to treat cancer. One such virus is vaccinia virus, which has been studied as a vector for delivering therapeutic genes to cancer cells, as an oncolytic virus that prolifer...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/768A61K38/20A61K47/18A61K47/26A61K9/08C12N7/01C12N15/24C12N15/39A61P35/00
CPCA61K38/2046A61K35/768A61K47/26A61K47/18A61K9/08A61K9/0019C12N7/00C07K14/5418C07K14/5434C07K14/475C07K14/005A61P35/00C12N2710/24021C12N2710/24022C12N2710/24032A61K38/208A61K2300/00A61P35/04A61K39/39558A61K45/06C12N15/86C12N2710/24121C12N2710/24132C12N2710/24143C12N2710/24162C12N2710/24171
Inventor 中尾慎典纲野伸明荒井幸规
Owner ASTELLAS PHARMA INC
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