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Tumor targeting recombinant newcastle disease viruses and construction method thereof

A Newcastle disease virus and tumor-targeting technology, which is applied in the direction of anti-tumor drugs, recombinant DNA technology, drug combination, etc., can solve the problems of reducing the dose of NDV finally binding to target tumor cells, coagulation and hemolysis, etc.

Active Publication Date: 2010-12-08
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This unfavorable factor causes NDV to adsorb to normal cells when NDV is injected intravenously, which undoubtedly reduces the dose of NDV that finally binds to target tumor cells.
On the other hand, we found that NDV binds to mouse or human erythrocytes, leading to severe coagulation and hemolysis

Method used

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  • Tumor targeting recombinant newcastle disease viruses and construction method thereof
  • Tumor targeting recombinant newcastle disease viruses and construction method thereof
  • Tumor targeting recombinant newcastle disease viruses and construction method thereof

Examples

Experimental program
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Embodiment Construction

[0022] 1 material

[0023] 1.1 Viruses and cell lines

[0024] The virulent strain NDV Italien (for reference, see "In vivo efficacy of systemic tumor targeting of a viral RNA vector with oncolytic properties using a bispecific adapter protein" Int J Oncol.2006Dec; 29(6): 1359-69.) was donated by Professor Schirrmacher; BSR T7 / 5 cells expressing T7 RNA polymerase were donated by Prof. K. Conzelmann of the Pettenkofer Institute in Munich, Germany.

[0025] 1.2 Strains and plasmids

[0026] E.coli JM109 was purchased from Beijing Dingguo Biotechnology Co., Ltd.; pTA2Vector was purchased from Toyobo Company; pBR322 vector was purchased from TaKaRa Company.

[0027] 1.3 Main reagents

[0028]Trizol LS Reagent and Lipofectamine TM2000 were purchased from Invitrogen; IPTG, X-gal, ampicillin, and G418 were purchased from Genview; ReverTra Ace reverse transcriptase was purchased from Toyobo; restriction enzymes, Ex taq HS enzyme, T4 DNA Ligase was purchased from TaKaRa Company; Pl...

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Abstract

The invention relates to a tumor-targeted recombinant Newcastle virus and a construction method thereof, wherein, 3'of an HN gene open reading frame of an Italien strain virus genome of the Newcastle virus is connected with a tumor scFv sequence through a G4S linker, and fusion expression of the two proteins can be realized; simultaneously, relative sequences which are responsible for coding sialic acid acceptors in HN genes are mutated, and the virus loses identification and combining capacities on sialic acids, and finally the recombinant virus can only be combined with tumor cells which express tumor associated antigens and replicated in the tumor cells which are then caused to be dead. The invention has the advantages that the recombinant oncolytic Newcastle virus which can be combined with the tumor cells which express the tumor associated antigens by means of targeting and replicated in the tumor cells which are finally killed, and is avirulent to normal human histiocytes. The invention provides a possible proposal for therapy research of associated tumors.

Description

technical field [0001] The invention relates to the fields of genetic engineering, biotechnology and medicine, in particular to construct a recombinant Newcastle disease virus capable of specifically infecting and killing tumor cells by recognizing and combining with antigens related to the surface of tumor cells and its construction method. Background technique [0002] Newcastle disease virus (NDV) is a negative-strand RNA virus that causes respiratory tract infection and other symptoms in poultry. Because NDV is non-pathogenic to humans and has the characteristics of selective replication in tumor cells, it has been used in the treatment of tumors in recent years. [0003] NDV is a non-segmented, negative-strand RNA paramyxovirus with a diameter of 100-400 nm, approximately spherical, and the nucleocapsid is surrounded by a double-layer lipid membrane derived from the cytoplasmic membrane. The total length of the RNA genome is about 15186 nucleotides, including 6 genes, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/86A61K48/00A61P35/00
Inventor 边惠洁陈志南尉丁李亚琳张华
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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