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Attenuated salmonella typhimurium and application of genetically engineered bacterium of attenuated salmonella typhimurium in preparation of medicine for treating prostatic cancer

A technology of Salmonella typhi and genetically engineered bacteria, applied in the field of medicine, can solve problems such as side effects and immune reactions, and achieve a significant inhibitory effect

Active Publication Date: 2014-08-06
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, often causing the body's immune response

Method used

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  • Attenuated salmonella typhimurium and application of genetically engineered bacterium of attenuated salmonella typhimurium in preparation of medicine for treating prostatic cancer
  • Attenuated salmonella typhimurium and application of genetically engineered bacterium of attenuated salmonella typhimurium in preparation of medicine for treating prostatic cancer
  • Attenuated salmonella typhimurium and application of genetically engineered bacterium of attenuated salmonella typhimurium in preparation of medicine for treating prostatic cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: the construction of genetically engineered bacteria.

[0030] (1) Construction of a plasmid expressing the L-methioninase gene.

[0031] 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:

[0032] The pSVSPORT plasmid was digested with Kpn I and Hind III. The digestion system was: 2 μg plasmid DNA, 3 μL 10×buffer, 1.5 μL Kpn I enzyme, 1.5 μL Hind III enzyme, added ddH2O to make up the volume to 30 μL, and incubated at 37 ° C for 3 h. 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.

[0033] The DNA fragment of the L-methioninase coding region obtained by whole gene syn...

Embodiment 2

[0042] Example 2: Antitumor effect of VNP20009 and its genetically engineered bacteria.

[0043] 1. Cultivate androgen-independent prostate cancer cell PC-3 with F-12K medium containing 10% fetal bovine serum, with a cell number of 2×10 6 Inoculated subcutaneously in the right armpit of nude mice. Every 2 to 3 days, the state of the mice was observed, and the size of the tumor was measured with a vernier caliper (volume=0.52×length×width) 2 ). When the tumor volume reaches 0.1-0.2cm 3 , the tumor-bearing nude mice were randomly divided into groups: PBS control group, VNP20009, VNP20009-V group and VNP20009-M group.

[0044] 2. Cultivate VNP20009, VNP20009-V and VNP20009-M with LB-O, when OD ≈ 0.6, collect the bacteria, then resuspend with PBS, dilute with 2×10 6 The dose of CFU / rat was administered by intratumoral injection, and the control group was injected with the same volume of PBS. After administration, the activity, food intake and body weight of the nude mice were...

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Abstract

The invention discloses attenuated salmonella typhimurium and application of a genetically engineered bacterium of the attenuated salmonella typhimurium in preparation of a medicine for treating the prostatic cancer. The attenuated salmonella typhimurium has a tumor targeting property and an obvious suppression effect on prostate cancer cells, and the genetically engineered bacterium consisting of the attenuated salmonella typhimurium and plasmids also has the tumor targeting property; furthermore, the attenuated salmonella typhimurium with the plasmids cloned with an L-methioninase gene can continuously express L-methioninase in a tumor tissue, and methionine and other nutritional substances are greatly consumed, so that tumor cells are lack of nutrition and grow slowly; therefore, the attenuated salmonella typhimurium can be used for preparing the medicine for treating the prostatic cancer.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to the application of attenuated Salmonella typhimurium and its genetically engineered bacteria in the preparation of drugs for treating prostate cancer. Background technique [0002] Prostate cancer is the most common malignant tumor in men in European and American countries. Its incidence rate ranks first in male malignant tumors in the United States, and its mortality rate is second only to lung cancer. In recent years, the incidence of prostate cancer in my country has also shown an increasing trend year by year, and the histological malignancy of prostate cancer is higher than that of American patients. According to the survey on the relative survival rate of patients with malignant urological tumors in Shanghai, China, 80.0% to 90.0% of patients in China have advanced prostate cancer when they see a doctor, and the 5-year survival rate is less than 30%. Due to the large popu...

Claims

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Application Information

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IPC IPC(8): A61K48/00C12N15/74C12N1/21A61P35/00C12R1/42
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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