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Application of genetically engineered bacterium VNP-20009-M in preparation of medicines for preventing and treating cancer metastasis

A technology of VNP20009-M and genetically engineered bacteria, applied in the field of medicine, can solve problems such as immune response and side effects, and achieve the effect of inhibiting invasion and metastasis and tumor metastasis

Pending Publication Date: 2016-10-05
GUANGZHOU SINOGEN PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since mammals themselves do not express methionase, the way of exogenous administration has certain side effects, which often cause the body's immune response

Method used

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  • Application of genetically engineered bacterium VNP-20009-M in preparation of medicines for preventing and treating cancer metastasis
  • Application of genetically engineered bacterium VNP-20009-M in preparation of medicines for preventing and treating cancer metastasis
  • Application of genetically engineered bacterium VNP-20009-M in preparation of medicines for preventing and treating cancer metastasis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1: the construction of genetically engineered bacteria.

[0050] (1) Construction of a plasmid expressing the L-methioninase gene.

[0051] Synthesized L-methioninase (GenBank: L43133.1) gene was subcloned into pUC57 plasmid (GenScript), and then subcloned into pSVSPORT plasmid (invitrogen) through Kpn I and Hind III restriction sites to obtain pSVSPORT-L- methioninase expression plasmid. The specific construction process is as follows:

[0052] Digest the pSVSPORT plasmid with Kpn I and Hind III, the enzyme digestion system is: 2 μg plasmid DNA, 3 μL 10× buffer, 1.5 μL Kpn I enzyme, 1.5 μL Hind III enzyme, add ddH 2 Make up the volume with O to 30 μL, and incubate at 37°C for 3h. Then the digestion system was separated by electrophoresis in 1% agarose gel, the DNA band with a size of 4.1 kb was cut out, and the DNA was purified with a gel recovery and purification kit.

[0053] The DNA fragment of the L-methioninase coding region obtained by whole gene s...

Embodiment 2

[0062] Example 2: Anti-tumor metastasis effect of genetically engineered bacteria VNP20009-M.

[0063] 1. The effect of genetically engineered bacteria VNP20009-M on the invasion ability of prostate cancer cells. Prostate cancer cells PC-3 (androgen-independent) in the logarithmic growth phase were inoculated in 6-well plates, and when the cell density grew to about 70%-80%, 1*10 6 Add VNP20009-V and VNP20009-M to CFU / well, and set up a blank control group. After bacteria and cells were co-cultured for 30 minutes, they were washed 5 times with PBS, and then the cells were digested with trypsin, collected by centrifugation, counted, and counted according to 5*10 4 / small chamber The cells were added to the transwell chamber covered with matrigel glue, the upper chamber used serum-free medium, and the lower chamber used the medium containing 20% ​​fetal bovine serum. Both PBS and medium contain 12 μg / mL penicillin, 20 μg / mL streptomycin, and 10 μg / mL kanamycin. After 36 hour...

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Abstract

The invention discloses application of genetically engineered bacterium VNP-20009-M of attenuated salmonella typhimurium in preparation of medicines for preventing and treating metastasized cancers. The genetically engineered bacterium VNP20009-M has a tumor targeting property on cancer cells, has a remarkable effect for inhibiting metastasis and growth of cancer cells, and can be used for preparing medicines for preventing and treating metastasized tumors.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to the application of genetically engineered bacteria VNP20009-M in the preparation of medicines for preventing and treating cancer. Background technique [0002] Malignant tumors are major diseases that threaten human health and patient survival in today's society. According to the statistics of the World Health Organization (WHO), in 2012, there were about 14 million new cancer patients worldwide, and about 8.2 million people died of cancer. According to the 2012 data released by my country Cancer Registry Center, there are about 3.5 million new cancer cases in China every year, and about 2.5 million people die from it. Among them, tumor metastasis is the leading cause of death in cancer patients. Tumor metastasis is a complex, multi-step biological process. Cancer cells in a patient's body develop to a certain extent and will migrate to distant places through the circulatory s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K38/51A61K38/00A61P35/00A61P35/04C12N1/21C12N15/74C12N15/60C12R1/42
CPCA61K38/005A61K38/51A61K48/005C12N9/88A61K35/74A61P35/04C12Y404/01011A61K48/00C12N15/74Y02A50/30C12R2001/42C12N1/205
Inventor 赵子建周素瑾林艳赵正刚李芳红
Owner GUANGZHOU SINOGEN PHARMA CO LTD
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