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Attenuated salmonella typhimurium and application of genetically engineered bacterium of attenuated salmonella typhimurium to preparation of medicines for treating pancreatic cancer

A technology of Salmonella typhi 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 slow growth and significant inhibitory effect

Active Publication Date: 2014-03-26
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 to preparation of medicines for treating pancreatic cancer
  • Attenuated salmonella typhimurium and application of genetically engineered bacterium of attenuated salmonella typhimurium to preparation of medicines for treating pancreatic cancer
  • Attenuated salmonella typhimurium and application of genetically engineered bacterium of attenuated salmonella typhimurium to preparation of medicines for treating pancreatic cancer

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1: the construction of genetically engineered bacteria.

[0025] (1) Construct a plasmid expressing L-methioninase gene.

[0026] 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:

[0027] Digest the pSVSPORT plasmid with Kpn I and Hind III. The 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, and a 4.1kb DNA band was cut out, and the DNA was purified with a gel recovery and purification kit.

[0028] The DNA fragment of the L-methioninase coding region obtained by whole gene synthesis was subcloned into the ...

Embodiment 2

[0037] Example 2: Antitumor effect of VNP20009 and its genetically engineered bacteria.

[0038] 1. CFPAC-1 pancreatic cancer cells were cultured in DMEM medium containing 10% fetal bovine serum, and the number of cells was 2×10 6 Inoculated subcutaneously in the right armpit of nude mice. Observe the state of the mice every 2 to 3 days, and measure the tumor size 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 group, VNP20009 group, VNP20009-V group and VNP20009-M group.

[0039] 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. Six days later, the same dose was administered for the second time. After administration, the activity, foo...

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PUM

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Abstract

The invention discloses attenuated salmonella typhimurium and application of a genetically engineered bacterium of the attenuated salmonella typhimurium to preparation of medicines for treating pancreatic cancer. The attenuated salmonella typhimurium has tumor targeting and a remarkable inhibiting effect on pancreatic cancer cells; the genetically engineered bacterium constructed by the attenuated salmonella typhimurium and a plasmid also has tumor targeting; the attenuated salmonella typhimurium with the plasmid with a clonal L-methioninase gene can continuously express L-methioninase on a tumor tissue to consume methionine and other nutrient substances to cause undernourishment and slow growth of tumor cells; therefore, both the attenuated salmonella typhimurium and the genetically engineered bacterium can be used for preparing the medicines for treating the pancreatic 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 pancreatic cancer. Background technique [0002] Pancreatic cancer is a digestive system tumor with insidious onset, rapid progression, poor prognosis and high degree of malignancy. The morbidity and mortality of pancreatic cancer in China are showing an increasing trend year by year, and the mortality of pancreatic cancer patients in men and women ranks sixth and seventh in all malignant tumors, respectively. According to reports, the relative 5-year survival rate of patients with pancreatic cancer is only 5.5%. At present, the treatment of pancreatic cancer mainly includes surgical treatment, adjuvant treatment, palliative and supportive treatment. Pancreatic tumors are often overlooked because the pancreas is hidden behind the stomach. At pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61P35/00C12N1/21C12N15/63C12R1/42
Inventor 林艳周素瑾赵子建李芳红赵正刚李晓曦于鹏丽
Owner GUANGZHOU SINOGEN PHARMA CO LTD
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