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Preparation and application of targeted hypersensitive broad-spectrum oncolytic virus

A recombinant virus, Newcastle disease virus technology, applied in the field of genetic engineering, can solve the problems of drugs with poor efficacy, unsatisfactory tissue-specific expression of gene drugs, poor targeting, etc., to solve the problems of poor efficacy and enhance anti-tumor immune response. Effect

Active Publication Date: 2021-11-26
旦旦生物科技(上海)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Previous patents reported that Newcastle disease virus NDV has weak lethality, anti-cancer effect is not ideal, poor targeting and tissue-specific expression of gene drugs are not ideal
One of the reasons for the poor efficacy of current solid tumor treatments is the inability of drugs to enter the tumor interior

Method used

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  • Preparation and application of targeted hypersensitive broad-spectrum oncolytic virus
  • Preparation and application of targeted hypersensitive broad-spectrum oncolytic virus
  • Preparation and application of targeted hypersensitive broad-spectrum oncolytic virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0123] Reagents required for the experiment:

[0124] 1. The large plasmid extraction kit is a product of Vazyme, the product number is DC202.

[0125] 2. DMEM is a product of Solarbio Company, the product number is 12100-500ml.

[0126] 3. G-418 is a product of Solarbio, the product number is G8161.

[0127] 4. Opti-MEM is a product of Gibco, the product number is 31985088.

[0128] 5. The transfection reagent is the product of Vazyme Company, the product number is T202-01.

[0129] 6. Cytarabine is a product of McLean, the product number is C805253-5g.

[0130] 7. TPCK is a product of Meilun Biological Company, the article number is MB3477.

[0131] 8. Day-old chicken embryos were purchased from Jinan Saisi Poultry Technology Co., Ltd.

[0132] 9. The RNA extraction kit is a product of Beijing Quanshijin Company, the article number is AT411-02.

[0133] 10. The RT-PCR kit is a product of Beijing Quanshijin Company, the article number is AT411-02.

[0134] experimental...

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Abstract

The invention discloses preparation and application of a targeted hypersensitive broad-spectrum oncolytic virus. The targeted hypersensitive broad-spectrum oncolytic virus is a recombinant virus containing a recombinant Newcastle disease virus, a hypersensitive gene alpha-1, 3GT, a heparanase gene HPSE for dissolving an extracellular matrix, and a FAP scFv gene (NDV-GT-HPSE-FAP scFv, called NDV-GHF for short) plasmid for targeting tumor-associated fibroblasts; and the recombinant virus is obtained by co-transfecting a BSR cell with a recombinant Newcastle disease virus NDV-GHF plasmid and a helper plasmid and rescuing. The recombinant Newcastle disease virus NDV-GHF plasmid is a recombinant Newcastle disease virus NDV-GHF plasmid obtained by recombining NDV with alpha-1, 3GT, HPSE and FAP scFv genes; the sequence of the HPSE gene is as shown in SEQ ID NO: 1; the sequence of the FAP scFv gene is as shown in SEQ ID NO: 2; and the sequence of the alpha-1, 3GT gene is as shown in SEQ ID NO: 3. The invention also discloses a preparation method of the recombinant virus and an application of the recombinant virus in melting a tumor physical barrier.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and relates to a targeted hypersensitive broad-spectrum oncolytic virus for melting tumor physical barriers and its preparation and application. Background technique [0002] Heparanase (HPSE) is the only endogenous glycosidase in mammalian cells that can cleave heparan sulfate proteoglycan side chains (heparan sulfate chains, HS) in the extracellular matrix (that is, cleave heparan sulfate protein at a specific site glycans). In recent years, studies have found that heparanase plays an important role in tumor immune surveillance. On the one hand, it activates natural killer cells (natural killer, NKC); on the other hand, it cuts HS chains and subsequent extracellular matrix (extracellular matrix, ECM) degrades and destroys the physical barrier in tumor tissue, allowing chemokines to enter the tumor stroma, attracting NK cells, T cells, and dendritic cells (dendritic cells, DC) to ga...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/86C12N15/40C12N15/56C12N15/13A61K35/768A61K48/00A61K38/47A61K39/395A61P35/00
CPCC12N7/00C12N9/2402C12N15/86C07K14/005C07K16/30A61K35/768A61K48/005A61K48/0008A61K38/47A61K39/39558A61P35/00C12Y302/01166C12N2760/18121C12N2760/18122C12N2760/18143C12N2760/18132A61K2039/5256A61K2039/54
Inventor 钟莉娉
Owner 旦旦生物科技(上海)有限公司
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