Herpes simplex virus vector, recombinant virus, host cell and pharmaceutical composition thereof

A technology of herpes simplex virus and virus vector, which is applied in the fields of medical formula, plant genetic improvement, genetic material composition, etc. It can solve the problems of low virus infection rate, slow replication and proliferation speed, etc., and achieve high infection rate, low toxicity, and dissolution powerful effect

Active Publication Date: 2009-03-04
北京奥源和力生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the genomes of herpes simplex virus type I that infect different races have significantly different restriction endonuclease decomposition patterns, that is, there are restriction fragment length polymorphisms (restriction fragment length polymorphism, RFLP) (see "Quantitative analysis of genomic polymorphism of herpes simplex virus type 1 strains from six countries: studies of molecular evolution and molecular epidemiology of the virus", H. Sakaoka et al., Journal of General Virology, Vol 75, 513-527, 1994), herpes simplex virus 17+ strains from white Therefore, one of the genes encoding cell-infecting polypeptide 34.5, the gene encoding cell-infecting polypeptide 6, and the gene encoding cell-infecting polypeptide 47 is knocked out of the type I herpes simplex virus 17+ strain that is susceptible to infecting white people or The vectors obtained after several times have the disadvantages of low virus infection rate and slow replication and proliferation speed in host cells when used for gene therapy of yellow race

Method used

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  • Herpes simplex virus vector, recombinant virus, host cell and pharmaceutical composition thereof
  • Herpes simplex virus vector, recombinant virus, host cell and pharmaceutical composition thereof
  • Herpes simplex virus vector, recombinant virus, host cell and pharmaceutical composition thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0196] This example illustrates the preparation of a herpes simplex virus vector of the present invention.

[0197] (1) Collection of virus

[0198] An adult Chinese male, 37 years old, Han nationality, with a history of repeated oral herpes, without other diseases, had a typical clinical manifestation of herpes simplex virus type I recurrent infection—the sample was taken two days after herpes-like blisters appeared around the corner of the mouth.

[0199] Take a 15 ml sterile centrifuge tube, which contains 3 ml of sterile phosphate buffered saline (PBS) containing 100 μg / ml kanamycin, which contains 9 g / L of NaCl, 0.21 g / ml of liters of KH 2 PO 4 and 0.97 g / L of Na 2 HPO 4 .12H 2 O: Squeeze the herpetic vesicle with a sterile cotton swab until the light yellow secretion flows out, then dip the secretion with a cotton swab, and rinse the end of the obtained cotton swab with the secretion in the above-mentioned PBS buffer for 10 seconds. Take out the cotton swab, and us...

Embodiment 2

[0211] This example illustrates the lytic ability of the herpes simplex virus vector of the present invention to human nasopharyngeal carcinoma cells (CNE-1) with multiple occurrences in yellow race.

[0212] (1) Determination of Herpes Simplex Virus Vector Titer

[0213] With DMEM medium containing 10% fetal bovine serum, 5 × 10 5 Inoculate African green monkey kidney cells (Vero cells) (ATCC, USA) at a density of 1 cell / well, at 37°C in 5% CO 2 Cultivate in the incubator for 24 hours until each well of Vero cells is 80% full and ready for use.

[0214] Under sterile conditions, collect the supernatant culture fluid of the Vero cell culture cultured with 7 kinds of herpes simplex virus vectors shown in Table 2 of Example 1 respectively, and use serum-free DMEM medium to gradiently dilute to 1 × 10 respectively. 2 Double solution, 1×10 3 Double solution, 1×10 4 Double solution, 1×10 5 Double solution, 1×10 6 Double solution and 1×10 7 100 microliters of each dilution. ...

Embodiment 3

[0234] This example demonstrates the lytic ability of the herpes simplex virus vector of the present invention to general tumor cells.

[0235] According to the method of Example 2, human breast cancer cell MCF-7 and human colon cancer cell HT-29 were transfected at two different multiplicity of infection (MOI=0.1 and MOI=1) respectively with the herpes simplex virus vector of the present invention. and human lung adenocarcinoma cell A549 (purchased from the Second Research Institute of the Academy of Military Medical Sciences), and the number of survival tumor cells at different time points after transfection was tested, and the results are shown in Table 6.

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Abstract

The invention provides a herpes simplex virus carrier which is separated from the yellow race and lacks one or more than one gene of the gene of coded infection cell multi-peptide 34.5, the gene of coded infection cell multi-peptide 6 and the gene of coded infection cell multi-peptide 47; the virus carrier has high specificity infection rate to the host cells of the yellow race with high reproduction speed and low toxicity, so the virus carrier can be directly used for injecting and dissolving the tumor cells of the yellow race, and carry genomic medicine into the cells of the yellow race to have gene therapy.

Description

technical field [0001] The present invention relates to a viral vector, a recombinant virus comprising the viral vector, a host cell comprising the viral vector and / or the recombinant virus, and a pharmaceutical composition comprising the viral vector and / or the recombinant virus . Specifically, the present invention relates to a herpes simplex virus vector, a recombinant virus comprising the viral vector, a host cell comprising the viral vector and / or the recombinant virus, and a host cell comprising the viral vector and / or the Pharmaceutical composition of recombinant virus. Background technique [0002] Herpes simplex virus (HSV) is spherical, and the complete herpes simplex virus consists of core, capsid, tegument and envelope. The core of the virus contains double-stranded DNA, wound into a filamentous scroll. The capsid is icosahedral, with a diameter of about 100 nanometers and consists of 162 shell particles. The capsid is covered by a membrane of uneven thicknes...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/869C12N15/33A61K48/00
Inventor 李秉珅李巍东田超李琦
Owner 北京奥源和力生物技术有限公司
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