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Application of expression TRAIL protein colon bacillus for preparing medicine to treat tumor

A technology for the treatment of Escherichia coli and tumors, which is applied in the direction of anti-tumor drugs, gene therapy, drug combinations, etc., to achieve broad market prospects and solve the effects of toxic effects

Inactive Publication Date: 2007-01-31
BIOMEDICAL ANALYSIS CENT OF ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] At present, there is no report of linking Escherichia coli with TRAIL, using the tumor targeting of Escherichia coli to bring TRAIL to the tumor site, and exerting the anti-tumor effect of TRAIL at a specific site

Method used

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  • Application of expression TRAIL protein colon bacillus for preparing medicine to treat tumor
  • Application of expression TRAIL protein colon bacillus for preparing medicine to treat tumor
  • Application of expression TRAIL protein colon bacillus for preparing medicine to treat tumor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Establishment of animal tumor model and metastatic tumor model

[0039] 1. Materials:

[0040] (1) Cells, bacteria and culture medium

[0041] Tumor cells B16-F10-luc-G5, MCF-7-luc-F5, BEL7405-luc and SMMC7721-luc were purchased from Xenogen Company, PC-3, C6, MB-49 and HT1080 were purchased from ATCC Company, E.coli DH5α , BL21 and JM109 are commercially available and kept in this laboratory. Cell culture medium: RPMI-1640, DMEM, and fetal bovine serum are products of Sigma Company. Bacterial culture medium: peptone and yeast extract of each component in LB were purchased from OXOID Company, and sodium chloride was domestic analytical grade.

[0042] (2) Plasmids, vectors and libraries

[0043] The pXen-1 (luxABCDE) plasmid was purchased from Xenogen Company, pGEX-KG, pET-28a(+) and pBV220 were purchased from the market and kept in our laboratory, and the human breast cDNA library was purchased from Clotech Company.

[0044] (3) Experimental anim...

Embodiment 2

[0064] Example 2 Construction and Detection of Luminescent Bacteria

[0065] 1. Materials

[0066] Same as Example 1.

[0067] 2. Methods

[0068] (1) Preparation of luminescent bacteria

[0069] Bacterial medium LB was prepared as usual, and competent cells such as Escherichia coli DH5α, BL21, and JM109 were prepared as usual.

[0070] The pXen-1 (luxABCDE) was transformed into Escherichia coli DH5α, BL21 and JM109 according to the conventional method, and the luminescence and efficiency of the clones were observed by IVIS imaging system after plating.

[0071] (2) Treatment of bacteria before the experiment

[0072] ① One day before injecting bacteria into mice, pick monoclonal colonies with higher luminescence efficiency and culture them in LB with Amp resistance overnight, transfer the bacteria at 1:100 the next day, and continue to shake until logarithmic growth The middle and late stage is OD600=0.6~1.0.

[0073] ②Press 1OD=8×10 8 Calculate the amount of b...

Embodiment 3

[0079] Example 3 Study on the distribution characteristics and tumor localization characteristics of bacteria in animals

[0080] 1. Materials:

[0081] Same as Example 1.

[0082] 2. Methods and Results

[0083] (1) Characteristics of bacteria distribution in normal mice

[0084] 2 x 10 intravenously 9 cfu / 100μl / mouse or 1×10 9 Within 10 to 20 minutes of cfu / 100μl / mouse bacteria in normal mice, the bacteria were distributed in the lungs, and then the bacteria gradually accumulated in the liver and localized in the liver (see appendix). figure 1 ). Most mice cannot tolerate greater than 1×10 9 Cfu / 100μl / mouse dose of bacteria, most mice died on the second day after injection, autopsy results showed that bacteria mainly accumulated in the lungs and livers of mice. The bacteria are only distributed in the liver. Intravenous 1×10 8 The mice with cfu / 100μl / mouse bacteria could all survive, and the distribution of bacteria was finally distributed only to the liver.

...

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Abstract

An application of the colibacillus expressing TRAIL protein in preparing the medicines for treating tumor is disclosed. Said colibacillus with tumor target and carrying said TRAIL protein can be located in the tumor tissue to make the TRAIL protein to play its stronger suppression role to tumor.

Description

technical field [0001] The invention relates to the medical use of bacteria, in particular to the use of Escherichia coli expressing TRAIL protein in the preparation of tumor therapeutic drugs. Background technique [0002] Tumor is a major disease that seriously threatens human health. At present, there are two major problems in tumor treatment: First, effective means and methods for tumor treatment. Traditional tumor treatment modes, such as surgical resection, radiotherapy, chemotherapy, etc., have the disadvantages of poor treatment effect and large side effects. With the development of molecular biology and genetic engineering, many new and effective treatment methods have emerged, such as monoclonal Application of antibodies, pro-apoptotic factors, angiogenesis inhibitors, cytokines and suicide genes. The emergence of these methods has brought new opportunities for tumor treatment, and at the same time, it has also brought new problems, that is, the second biggest pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K47/00A61K35/74C12N1/21C12N15/63A61P35/00
Inventor 张学敏张海英梁冰
Owner BIOMEDICAL ANALYSIS CENT OF ACADEMY OF MILITARY MEDICAL SCI
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