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Oncolytic virus preparation and preparing method thereof

An oncolytic virus and preparation technology, applied in biochemical equipment and methods, viruses, viruses/phages, etc., can solve the problems of normal tissue damage, toxic side effects, weakening the effective killing effect of viruses on tumor cells, etc., to improve tumor killing. effect, the effect of avoiding toxic side effects

Inactive Publication Date: 2015-10-07
HUBEI SOUNDNY BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as an alien component, oncolytic virus will be rejected by the body once it enters the body, especially the production of anti-viral antibodies, which will limit the anti-tumor effect of oncolytic virus by clearing the virus
In addition, free oncolytic virus easily enters normal tissues, which not only weakens the effective killing effect of the virus on tumor cells, but also causes damage to normal tissues, causing toxic side effects

Method used

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  • Oncolytic virus preparation and preparing method thereof
  • Oncolytic virus preparation and preparing method thereof
  • Oncolytic virus preparation and preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Using oncolytic adenovirus to induce apoptosis of human lung cancer cells to produce microparticles

[0044] 1. Experimental steps

[0045] A549 human lung cancer cells were cultured in DMEM cell culture medium to make the number of cells reach 1×10 7 ;Add 1×10 8 Oncolytic adenovirus was used to infect human lung cancer cells; the infected tumor cells were cultured at 37°C and 5% oxygen content, and at 48 hours after administration of oncolytic adenovirus, when the tumor cells shrank and darkened, the The supernatant of the human lung cancer cell culture medium was centrifuged at 1000g and 5000g for 10 minutes to remove cells and debris, and the centrifuged supernatant was further centrifuged at 10000g for 2 hours to collect the precipitate and obtain the A549 derived from apoptosis. Microparticles formed by encapsulation of oncolytic adenovirus in vesicles of lung cancer cells.

[0046] 2. Experimental results

[0047] The microparticles prepared above w...

Embodiment 2

[0048] Example 2: Oncolytic adenovirus induces apoptosis of human lung cancer cells. Cell vesicles produced encapsulate oncolytic adenovirus DNA.

[0049] 1. Experimental steps

[0050] Culture A549 human lung cancer cells to make the number of cells reach 1×10 7 ; Add 1×10 8 Oncolytic adenovirus particles; 48 hours after the administration of oncolytic adenovirus, when the tumor cells shrank and darkened, the supernatant of the human lung cancer cell culture medium was collected according to the steps in Example 1 to collect A549 human lung cancer cell apoptosis Microparticles produced (formed by encapsulation of oncolytic adenovirus in vesicles derived from apoptotic A549 human lung cancer cells).

[0051] On the one hand, the microparticles are treated with proteinase K to rupture and lyse the DNA molecules, and at the same time, the oncolytic adenoviruses are treated to separate and prepare the DNA molecules contained therein.

[0052] On the other hand, the micropartic...

Embodiment 3

[0058] A549 human lung cancer cells were cultured in DMEM cell culture medium to make the number of cells reach 1×10 7 ;Add 1×10 7 Oncolytic adenovirus was used to infect human lung cancer cells; the infected tumor cells were cultured at 37°C and 5% oxygen content, and at 48 hours after administration of oncolytic adenovirus, when the tumor cells shrank and darkened, the The supernatant of the human lung cancer cell culture medium was centrifuged at 1000g and 5000g for 10 minutes to remove cells and debris, and the centrifuged supernatant was further centrifuged at 10000g for 2 hours to collect the precipitate and obtain the A549 derived from apoptosis. Microparticles formed by encapsulation of oncolytic adenovirus in vesicles of lung cancer cells.

[0059] The microparticles prepared above were resuspended with 1ml of 0.9% (g / ml) physiological saline and then smeared, the microparticles were observed under an electron microscope, and the particle size of the microparticles w...

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Abstract

The invention provides an oncolytic virus preparation and a preparing method thereof. The oncolytic virus preparation comprises cell vesicles coming from apoptotic tumor cells and oncolytic viruses which are wrapped in the cell vesicles and serve as effective constituents. According to the oncolytic virus preparation, as the cell vesicles coming from the cells are used for wrapping the oncolytic viruses, the oncolytic viruses escape from attacks of an organism immune system and can reach the tumor treatment positions in a targeted manner, and the tumor killing effect is improved.

Description

technical field [0001] The invention relates to an oncolytic virus preparation and a preparation method thereof, in particular to an oncolytic virus preparation and a preparation method thereof. Background technique [0002] As a disease that seriously threatens human life and health, malignant tumors have become the top priority of medical research in recent years to find effective and less toxic treatment methods. According to the principle of using fire to fight fire, using virus to kill tumor cells is an ideal strategy for tumor treatment. However, this strategy faces two problems, namely, how to make the virus selectively reach the tumor site, and how to make the virus escape the body Attack of the immune system. [0003] Viral DNA or RNA is assembled with proteins and other biomolecules to form virus particles. If the latter can infect tumor cells and replicate in large numbers in tumor cells, new virus particles will be generated, which will eventually lead to tumor ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/761A61P35/00
CPCC12N7/00A61K35/761C12N2710/10351C12N2710/10332A61P35/00A61K9/5068A61K9/5089A61K9/0019A61K35/76A61K35/13C12N7/02C12N2720/12032C12N2760/20232C12N2760/20251
Inventor 黄波
Owner HUBEI SOUNDNY BIOLOGICAL TECH
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