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Application of benzethonium chloride in preparation of drugs to prevent and treat lung cancer

A technology of benzethonium chloride and medicine, which is applied in the field of medicine, can solve the problems of inhibiting the growth of lung cancer cells, and achieve the effects of small toxic and side effects, high safety and low price

Active Publication Date: 2019-05-24
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for lung cancer, there is no relevant literature and patent report that benzethonium chloride has an inhibitory effect on the growth of lung cancer cells

Method used

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  • Application of benzethonium chloride in preparation of drugs to prevent and treat lung cancer
  • Application of benzethonium chloride in preparation of drugs to prevent and treat lung cancer
  • Application of benzethonium chloride in preparation of drugs to prevent and treat lung cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1W

[0028] Example 1 WST-1 cell activity detection

[0029] 1. Cell plating: A549 and H1299 cells (purchased from the American Type Culture Collection (ATCC) cell bank) were collected, counted, plated in a 96-well plate at an amount of 3000 cells per well, and cultured overnight.

[0030] 2. Drug treatment: Dilute benzethonium chloride with DMEM complete medium to the treatment concentration (0, 5, 10, 15, 20 μM), then replace the original medium in the 96-well plate, and treat for 24, 48 and 72 hours respectively.

[0031] 3. WST-1 detection: according to the dilution ratio of 1:10 (w / v), dilute WST-1 (WST-1 cell proliferation detection kit) with DMEM complete medium to obtain the medium containing WST-1; then The medium containing WST-1 was used to replace the original medium in the 96-well plate, and the medium containing WST-1 was added to wells without cultured cells as a blank control, 100 μL per well. Then put the 96-well plate in a constant temperature incubator at 37°C a...

Embodiment 2

[0033] Example 2 Apoptosis

[0034] 1. Drug treatment: A549 and H1299 cells were collected and spread in 6-well plates, so that the cell density was about 40%, and DMEM medium containing 0, 10, 15 and 20 μM benzethonium chloride was added to treat the cells.

[0035] 2. Collect cells: After 48 hours of drug treatment, first collect the apoptotic cells in the supernatant, centrifuge at 1100rpm for 3 minutes, discard the supernatant, then digest the adherent cells with 0.25% trypsin, and collect them in the same centrifuge tube. After centrifugation at 1100rpm for 3min, the supernatant was discarded. Staining was performed after washing twice with PBS buffer.

[0036] 3. Staining: First resuspend the cells with 500 μL of Binding Buffer (Apoptosis Detection Kit), then add 5 μL of Annexin V-FITC to each tube and mix well, then add 5 μL of Propidium Iodide and mix well, then react for 15 minutes at room temperature in the dark .

[0037] 4. Flow cytometry: After staining, use a ...

Embodiment 3

[0039] Embodiment 3 subcutaneous tumor formation experiment in nude mice

[0040] 1. The animal experiments in this project strictly complied with the guidelines of animal care institutions, and all were approved by the Animal Experiment Ethics Committee of Jinan University.

[0041] 2. The experimental animals used in this project are 6-8 weeks old female nude mice (Balb / c-nu, purchased from the Experimental Animal Center of Southern Medical University), a total of 15, including 5 in the control group and 10 in the experimental group, Construct subcutaneous tumor formation model in nude mice:

[0042] (1) Cell collection: collect cells for cell counting, and use 1×10 cells per nude mouse 6 The total number of cells was calculated based on the amount of A549 cells, and the cells were resuspended in PBS buffer and mixed with Matrigel (purchased from Guangzhou Sier Biotechnology Co., Ltd., coring) at a volume ratio of 1:1.

[0043] (2) Inoculation: after the mice were anesthet...

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PUM

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Abstract

The invention discloses application of benzethonium chloride in the preparation of drugs to prevent and treat lung cancer. In-vitro and in-vivo experiments show that benzethonium chloride can inhibitthe growth and proliferation of lung cancer; since benzethonium chloride has little side effect, high safety and low price, benzethonium chloride may act a drug to treat lung cancer. In addition, it is also discovered herein that the joint use of benzethonium chloride and gefitinib can inhibit the growth of lung cancer cells more significantly than the sole use of gefitinib; it is indicated that the addition of benzethonium chloride can improve the sensitivity of lung cancer cells to gefitinib. Therefore, benzethonium chloride can be used as a gefitinib sensitizer, and alternatively, the combination of benzethonium chloride and gefitinib is applied to the treatment of lung cancer.

Description

technical field [0001] The invention belongs to the technical field of medicine, in particular to the application of benzethonium chloride in the preparation of medicines for preventing and treating lung cancer. Background technique [0002] Lung cancer is one of the most common malignant tumors and one of the most difficult diseases to treat in the world. In recent years, the incidence has shown an increasing trend. Because it is difficult to distinguish lung cancer from benign lung diseases, and the early symptoms of lung cancer are not obvious, most patients have developed to the middle and late stages when they seek treatment, so they often miss the best treatment period, which seriously affects the treatment effect. At present, lung cancer treatment methods include surgery, radiotherapy, chemotherapy, immunotherapy, traditional Chinese medicine, etc. However, the resistance of cancer to radiotherapy and chemotherapy leads to postoperative recurrence and poor prognosis o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/14A61P35/00A61K31/5377A61K9/08
Inventor 何庆瑜李斌黄小惠
Owner JINAN UNIVERSITY
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