Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Conditional duplicated adenovirus and its establishment and use

An adenovirus, replication-type technology, applied in the field of biomedicine, can solve problems such as not particularly strict control, virus replication, and limited application prospects of viruses

Inactive Publication Date: 2003-06-18
李川源 +1
View PDF0 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This greatly limits the application prospects of these viruses
In addition, the control of many promoters is not particularly strict, which will cause a certain amount of virus replication in non-targeted normal cells, thus narrowing the therapeutic window

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Conditional duplicated adenovirus and its establishment and use
  • Conditional duplicated adenovirus and its establishment and use
  • Conditional duplicated adenovirus and its establishment and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Isolation of Tumor-Specific Promoter A. Telomerase 5' Regulatory Sequence by PCR Reaction and Cloning

[0022] During natural DNA replication, the ends of chromosomes shorten with each cell division. The integrity of telomeres at the end of chromosomes is crucial to the stability of chromosomes. The integrity of telomeres is maintained by telomerase. Telomerase is inactive in more than 99% of normal cells and is reactivated in more than 90% of tumor cells. Experiments have shown that the activity of telomerase is controlled by its promoter. The promoter of telomerase (TERT) is active in most tumors but not in normal cells. In this way, the promoter of TERT becomes a tumor-selectively activated promoter. In order to isolate the telomerase promoter, the present invention adopts PCR technology, uses human cell DNA as a template, amplifies the promoter sequence 1, inserts it in the plasmid pEGFP-1 (purchased from Clontech Company), drives GFP gene expression, constructs...

Embodiment 2

[0028] Construction of tumor-specific replication-competent adenoviral vectors

[0029] The tumor-specific promoter isolated in Example 1 was embedded into the DNA of the adenovirus by genetic engineering, and used to control the expression of early genes necessary for the replication of the adenovirus. figure 1 Described is the construction method of recombinant virus. The virus is characterized in that their E1A and E4 genes are simultaneously regulated by tumor tissue-specific promoters. The promoters controlling E1A and E4 may be the same promoter or different promoters. The following combinations are combinations of conditionally replicative adenoviruses employed in the present invention.

[0030] 1) AdHRPE1aTERTE4-dsRed2, TSP1 is a hypoxia-inducible promoter, and TSP2 is a telomerase promoter;

[0031] 2) AdAFPE1aTERTE4-dsRed2, TSP1 is the α-embryoprotein promoter, and TSP2 is the telomerase promoter;

[0032] 3) AdE2F1E1aTERTE4-dsRed2, TSP1 is E2F1 promoter, TSP2 is...

Embodiment 3

[0037] Packaging, amplification, purification of virus, and characterization of infection and replication

[0038] Human embryonic kidney 293 cells were used to package the virus constructed above, and then amplified after screening by plaque purification, and the virus was purified by cesium chloride density gradient centrifugation and dialysis. The titer of purified virus can reach 3~5×10 10 pfu / ml. Observe virus replication in various tumor and normal cells. The result is as follows:

[0039] 1) AdTERTE1a-E4-dsRed2 can infect the following tumor cells, such as breast cancer, prostate cancer, ovarian cancer, cervical cancer, colon cancer, etc., and replicate in large numbers in these cells. Obvious cytopathic changes appeared 3-5 days after infection, and then the cells lysed and continued to infect surrounding tumor cells, and almost all tumor cells were killed within 3-7 days. The virus can also infect human fibroblasts and normal epithelial cells, but does not replica...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

A conditionally replicated adenovirus for treating tumor can be replicated selectively in tumor cell, but not nomral cell. Its two or more promotors of early gene needed by replication are respectively substituted by the promotor of tumor cell (or tissue). Said virus can effectively kill the tumor cells by the cracking of its cells or carrying the therapeutic gene. Its advantages are high curative effect and low by-effect.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to an adenovirus used for tumor gene therapy and its construction method and application. In particular, it relates to an adenovirus that can be selectively replicated in tumor cells but not replicated in normal cells, and is used for tumor gene therapy and its construction method and application Background technique [0002] In recent years, surgical treatment, chemotherapy and radiotherapy of tumors have been significantly improved, and the survival period of cancer patients has gradually increased. However, these improvements are still only applicable to early and localized lesions, and the "therapeutic window" between normal and tumor cells is still very narrow and difficult to widen. While these methods kill tumor cells, they also cause serious damage to normal cells of the patient, especially the hematopoietic and immune systems. Therefore, a considerable portion of tumors are still...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61P35/00C12N7/01C12N15/12C12N15/33C12N15/861
Inventor 李川源黄倩
Owner 李川源
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com