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Recombinant adenovirus for expressing human CREG and uses thereof

A recombinant adenovirus and application technology, applied in the field of recombinant adenovirus, can solve the problems of lack of clinical application feasibility, destruction of host cell genome stability, and no feasibility of clinical drug use.

Inactive Publication Date: 2009-07-08
GENERAL HOSPITAL OF THE NORTHERN WAR ZONE OF THE CHINESE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, retroviruses have the following defects: first, retroviruses can only infect vascular smooth muscle cells in the proliferative phase, and the infection efficiency is low; Therefore, it may destroy the stability of the host cell genome, and it is not feasible for clinical use; thirdly, in the study of preventing and treating restenosis after PCI, the most ideal and feasible route of administration is intravascular administration, while Retroviral vectors are only suitable for slow-release through the polyether (pluronic F127) carrier-wrapped vascular adventitia to infiltrate the vascular intima after arterial balloon injury (because the proliferation of vascular smooth muscle cells after balloon injury requires a certain process, intravascular immediate administration is ineffective), but this sustained-release method is only suitable for experimental studies on animals and lacks the feasibility of clinical application

Method used

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  • Recombinant adenovirus for expressing human CREG and uses thereof
  • Recombinant adenovirus for expressing human CREG and uses thereof
  • Recombinant adenovirus for expressing human CREG and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0032] Embodiment: 1. Construction of the recombinant adenoviral vector carrying the full sequence of human CREG cDNA

[0033] Design the RT-PCR primers of CREG gene according to the CREG cDNA sequence given in GenBank (NM_003851), and the primer sequences are as follows:

[0034] Forward primer: 5`---aa ggatcc atg gcc ggg cta tcc cgc-3'

[0035]Reverse primer: 5′-gc gaattc tca ctg aac tgt gac att ata ata ttcttc tgg-3′

[0036] Specific conditions:

[0037] dH 2 O 36.5 μL

[0038] dNTP 4μL

[0039] Buffer 5 μL

[0040] Primer (1) 1.5 μL

[0041] Primer (2) 1 μL

[0042] Template 1 μL

[0043] DNA polymerase (Takara company) 1 μL

[0044]

[0045] The mRNA was extracted from VSMC cultured in vitro and reverse-transcribed into cDNA (cDNA first-strand synthesis kit from TakaRa Company). Amplification conditions were: 25°C for 5 minutes, 42°C for 50 minutes, and 95°C for 10 minutes. A total of 667 bases of the human CREG cDNA coding sequence was amplified by the foll...

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Abstract

The present invention relates to a recombinant adenovirus of human E1A activating gene repressor (CREG) genes. The invention also relates to the use of the recombinant adenovirus for the preparation of drug having a clear inhibitory effect on the restenosis after PCI.

Description

technical field [0001] The invention relates to a recombinant adenovirus carrying a human E1A activator gene repressor (CREG) gene, and also relates to the use of the recombinant adenovirus for preparing a drug with definite inhibitory effect on post-PCI restenosis. Background technique [0002] Percutaneous coronary intervention (PCI) is currently one of the main means of coronary heart disease treatment. More than 2.8 million patients with coronary heart disease worldwide receive PCI treatment every year, but 10% of patients after surgery - The 70% incidence of restenosis reduces its benefit. In my country, the number of PCI is increasing year by year, and the number of restenosis is also increasing year by year, which has become a very serious problem. In view of the occurrence of restenosis, many methods have been used clinically to prevent its occurrence and development, including antithrombotic, lipid-lowering, etc. , PCI again, stent implantation, cutting balloon, dru...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/861C12N15/12A61K48/00A61P9/00
Inventor 韩雅玲郭亮邓捷康建
Owner GENERAL HOSPITAL OF THE NORTHERN WAR ZONE OF THE CHINESE PEOPLES LIBERATION ARMY
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