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Screening method of medicament for preventing prostatic cancer and application of nitazoxanide in pharmacy

A prostate cancer and screening method technology, applied in the field of biomedicine, can solve the problems of poor prognosis in patients with bone metastases, and achieve the effect of low acquisition cost

Pending Publication Date: 2022-03-04
SOUTH UNIVERSITY OF SCIENCE AND TECHNOLOGY OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, docetaxel and cabazitaxel are the only chemotherapeutic drugs with survival benefit for mCRPC patients, but virtually all mCRPC will eventually develop drug resistance, and the prognosis of patients with bone metastases treated with docetaxel is still poor

Method used

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  • Screening method of medicament for preventing prostatic cancer and application of nitazoxanide in pharmacy
  • Screening method of medicament for preventing prostatic cancer and application of nitazoxanide in pharmacy
  • Screening method of medicament for preventing prostatic cancer and application of nitazoxanide in pharmacy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1 Screening of Agents for Preventing Prostate Cancer

[0076] 1. Construction of PC3-KQ cells with high metastases and PC3-KR cells with low metastases in prostate cancer

[0077] 1. Conversion

[0078] Take the competent Escherichia coli out of the refrigerator at -80°C, and put it on ice to melt; add the ligation products (K369R, K369Q) constructed in the laboratory into the competent Escherichia coli, and incubate on ice; after heat shock at 42°C for 90s, place Place on ice for 2 minutes; add 200 μL of LB medium and 1 μL of antibiotics to the competent cells in a super-clean bench, pipette evenly, and then spread on a plate containing antibiotics; after coating, culture in a constant temperature incubator at 37°C overnight.

[0079] 2. Plasmid amplification

[0080] Use a 10 μL pipette tip to pick up the monoclonal colonies on the plate and put them into EP tubes. Shake the bacteria in a shaker at 200 rpm for 12 hours, and extract the plasmids according to ...

Embodiment 2

[0108] Example 2 Prostate Cancer Bone Metastasis Model Establishment

[0109] 1. Lentivirus infection to construct luciferase cells

[0110] 1. Explore the optimal concentration of puromycin (puromycin)

[0111] Before screening, it is necessary to explore the lowest concentration of puromycin that can kill empty cells: the cells can be plated in 24-well plates with a density of 5×10 per well. 4 After 24 hours, replace the complete medium with different concentrations of puromycin, the concentration of puromycin can be set to 0, 0.2, 0.4, 0.6, 0.8, 1.0, 2.0, 4.0, 6.0, 8.0, 10.0 mg / mL for 48 hours, select the one that can kill Subsequent experiments were performed at the lowest concentration of more than 90% empty cells.

[0112] 2. Lentivirus infection of cells

[0113] Day1: When the confluence of the cells is 80-90%, the cells are collected and spread on a 6-well plate, 1.5×10 per well 5 Generally, it is ensured that the cell confluence rate is between 30% and 50% when t...

Embodiment 3

[0126] Example 3 The preventive effect of nitazoxanide on prostate cancer bone metastasis

[0127] 1. In vivo imaging of small animals and microCT

[0128] In vivo imaging in mice: In vivo imaging was performed 7 days, 14 days, 21 days, 28 days, and 35 days after injection of PC3-KQ-LUC cells. The specific operation is as follows: 10 minutes before the imaging, start the in vivo imaging system, turn on the oxygen switch and the gas anesthesia switch, and then intraperitoneally inject the luciferase substrate (D-luciferin, SodiumSalt) prepared before the experiment into each group of mice, D- The concentration of the fluorescein working solution is 15 mg / mL, and the injection volume for each mouse depends on the weight of the mouse: 150 mg / kg; 10 minutes after injection into the mouse, the mouse is anesthetized for imaging analysis.

[0129] In addition, after the injection of PC3-KQ-LUC cells, nitazoxanide was administered immediately, and the administration method was intrag...

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Abstract

The invention discloses a screening method of a medicament for preventing prostate cancer and application of nitazoxanide in pharmacy, and the screening method comprises the following steps: S10, performing 3D cell culture on prostate cancer cells to obtain tumor spheroids; s20, a to-be-screened medicament is prepared into a solution with the concentration of 10 mu M, the solution is used for carrying out a cell invasion experiment on the tumor spheroids, the invasion area of the tumor spheroids is calculated, the medicament corresponding to the tumor spheroids with the invasion area smaller than 50% is screened out, and the to-be-screened medicament is selected from FDA marketing and pharmacopoeia recorded compound libraries; s30, preparing the screened medicaments into a plurality of solutions with different concentration gradients of 0-10 [mu] M, respectively adding the solutions into the tumor spheroids for culture, calculating the invasion areas of the tumor spheroids, and screening seven medicaments with the invasion areas less than 70%; and S40, measuring the IC50 of the seven medicaments, and screening out the medicament with the IC50 value less than 28 mu M, namely the medicament for preventing prostatic cancer. The nitazoxanide is good in safety and obvious in prevention effect.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a screening method of a medicament for preventing prostate cancer and the application of nitazoxanide in pharmacy. Background technique [0002] Prostate cancer is the most common malignant tumor of the male urinary system, and its incidence rate ranks fourth in the world. It is estimated that in 2020, 1,414,259 patients will develop prostate cancer, and 375,304 cancer patients will die from prostate cancer. Compared with European and American countries, although the incidence rate of prostate cancer in my country is relatively low, in recent years, with the aging of my country's social population, the improvement of economic level and the change of people's eating habits, the incidence rate of prostate cancer in my country is increasing year by year, which seriously threatens our country. Health of middle-aged and older men. According to the analysis of the current situatio...

Claims

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Application Information

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IPC IPC(8): C12Q1/02A61K31/426A61P35/00A61P35/04
CPCG01N33/5011A61K31/426A61P35/00A61P35/04G01N2500/10
Inventor 董金堂黄青青
Owner SOUTH UNIVERSITY OF SCIENCE AND TECHNOLOGY OF CHINA
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