Recombinant herpes simplex virus, host cell infected with recombinant herpes simplex virus, host cell for preparation of recombinant herpes simplex virus and their applications

A herpes simplex virus and cell-infecting technology, which is applied to cells modified by introducing foreign genetic material, virus/bacteriophage, and drug combinations, etc., can solve the problems of cross-contamination, immunogenicity, and differences in efficacy of mouse-derived viruses. , to avoid dependence and related side effects, long duration of treatment, and avoid drug diffusion

Inactive Publication Date: 2016-01-06
BEIJING WELLGENE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Mouse NGF has the following disadvantages: (1) there is a 10% difference in the protein sequence between mouse NGF and human NGF, the curative effect may be different, and it is immunogenic, which may induce antibody response; (2) the hidden danger of cross-contamination of mouse-derived virus; ( 3) Production depends on live animals
Due to the difficulty in extraction or the defect of recombination process, human nerve growth factor has not been released into the market

Method used

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  • Recombinant herpes simplex virus, host cell infected with recombinant herpes simplex virus, host cell for preparation of recombinant herpes simplex virus and their applications
  • Recombinant herpes simplex virus, host cell infected with recombinant herpes simplex virus, host cell for preparation of recombinant herpes simplex virus and their applications
  • Recombinant herpes simplex virus, host cell infected with recombinant herpes simplex virus, host cell for preparation of recombinant herpes simplex virus and their applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0027] According to one embodiment of the present invention, the target gene can directly utilize the LAP1 and LAP2 promoters in the LAT region without adding a promoter.

[0028] According to the present invention, in order to ensure that the target gene can be effectively translated independently, the target gene of the present invention preferably also includes a promoter, a start codon and a stop codon. In such a preferred situation, the recombinant herpes simplex When a virus infects a host cell and expresses its own gene, it can transcribe an independent and complete target mRNA segment, thereby effectively and independently translating the target gene.

[0029]According to the present invention, the promoter can be various conventional promoters in the art. According to a preferred embodiment of the present invention, the insertion site of the target gene includes insertion into the latency-related transcription coding region of the herpes simplex virus vector, and when...

Embodiment 1

[0050] This example is used to illustrate the construction of the recombinant herpes simplex virus of the present invention

[0051] According to the application number 2004100064921, the method recorded in the patent application of the authorized announcement number CN1283803C will knock out the ICP34.5 gene and the ICP4 gene of the wild-type clinically isolated HSV-1 virus, and in the latent expression region (LAT region) of the HSV-1 vector The 119499bp-122025bp downstream of the promoter was inserted into the ENK gene carrying the CMV promoter. Sequencing at Beijing Sanbo Yuanzhi Company confirmed that the ENK gene was correctly inserted into the herpes simplex virus vector. The successfully constructed viral vector was incubated at 37°C, 5% CO on the constructed Vero host cell line capable of expressing the ICP4 gene. 2 The multiplication of infection was 0.1. After harvesting, the cell debris was removed with a 0.65 μm filter, and then purified by high-speed centrifugat...

Embodiment 2

[0053] This example is used to illustrate the construction of the recombinant herpes simplex virus of the present invention

[0054] The construction of the recombinant herpes simplex virus was carried out according to the method in Example 1, except that the genes knocked out were the ICP34.5, ICP27 and ICP4 genes of the HSV-1 virus.

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Abstract

The invention relates to the field of biological medicines and discloses a recombinant herpes simplex virus. The recombinant herpes simplex virus comprises a herpes simplex virus vector and a target gene; the herpes simplex virus vector is obtained by enabling deficiency of an infection cell polypeptide 34.5 coding gene and an infection cell polypeptide 4 coding gene and selectable deficiency of an infection cell polypeptide 27 coding gene of a herpes simplex virus; the target gene is inserted in a latency associated transcript coding region and at a downstream position of a promoter of the coding region. The invention further provides a host cell infected with the recombinant herpes simplex virus, a host cell for preparation of the recombinant herpes simplex virus and applications of the recombinant herpes simplex virus, the host cell infected with the recombinant herpes simplex virus and the host cell for preparation of the recombinant herpes simplex virus in preparation of medicines for treatment of chronic pain, nerve injuries and/or drug addiction.

Description

technical field [0001] The invention relates to a recombinant virus, a host cell for preparing the recombinant virus, a host cell infected with the virus, and applications of the recombinant virus, a host cell for preparing the recombinant virus, and a host cell infected with the virus. Specifically, the present invention relates to a recombinant herpes simplex virus, a host cell for preparing the recombinant herpes simplex virus, a host cell infected with the recombinant herpes simplex virus, the recombinant herpes simplex virus, and a method for preparing the recombinant herpes simplex virus Use of host cells and host cells infected with the recombinant herpes simplex virus. Background technique [0002] Gene therapy is a revolution in the treatment of modern biomedicine, and carrier technology is the key to the success of gene therapy. Herpes simplex virus type 1 (HSV-1) vectors have a broad spectrum of host cells, can infect a variety of cells and continuously express t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N5/10A61K38/08A61K38/18A61P29/00A61P25/00A61P25/36
Inventor 于耕李劲风
Owner BEIJING WELLGENE CO LTD
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