Application of benzodihydropyran-containing spirooxindole compound in development of antitumor drugs and pharmaceutical composition of benzodihydropyran-containing spirooxindole
A kind of technology of chroman spiro and indole compound, which is applied in the field of anti-tumor drug development
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Embodiment 1
[0042] Example 1: In vitro anti-tumor activity evaluation of compounds
[0043]Tested tumor cells: human liver cancer cells HepG2, human lung cancer cells A549, human lung cancer cells H460, human prostate cancer cells DU145, human colon cancer cells HCT116, human colon cancer cells HCT15, human bladder cancer cells T24, human gastric cancer cells SGC-7901 , cisplatin-resistant human lung cancer cells (A549 / DDP), oxaliplatin-resistant human colon cancer cells (HCT116 / L), and 5-fluorouracil-resistant human colon cancer cells (HCT15 / 5-FU).
[0044] Cell culture: DMEM, DMEM-F12, MEM, RPMI 1640 and McCoys5A medium, 10% fetal bovine serum and 1% penicillin-streptomycin were prepared as culture medium. Cultured cell lines were placed at 37°C, 5% CO 2 The cells were routinely cultured and subcultured under saturated humidity, and the cells in the logarithmic growth phase were used in the experiments.
[0045] In vitro anti-tumor activity evaluation (MTT method):
[0046] The above...
Embodiment 2
[0053] Example 2: Evaluation of tumor cell apoptosis activity caused by compound 4
[0054] The applicant further carried out the compound 4-induced tumor cell apoptosis test, the test conditions are as follows: DU145, A549, and HepG2 cells in the logarithmic growth phase were plated in 6-well plates, and cells containing 10 μM compound 4 were added after the monolayer was overgrown. Incubate in maintenance solution, and set the cell control without drug treatment. After 48 hours, collect the cells, and use the Annexin-FITC / PI apoptosis detection kit to detect cell apoptosis on a flow cytometer.
[0055] Experimental results such as figure 1 It is shown that 10 μM compound 4 can potently induce the apoptosis of the above detected tumor cells.
Embodiment 3
[0056] Example 3: Evaluation of tumor cell G2 / M activity induced by compound 4
[0057] The applicant further implemented the compound 4-induced tumor cell apoptosis test, the test conditions are as follows: DU145, A549, and HepG2 cells in the logarithmic growth phase were plated in 6-well plates, and after the monolayer was overgrown, 2.5 μM compound 4 was added. Cells were incubated in maintenance solution, and a control cell without drug treatment was set. After 24 hours, the cells were collected, and the cell cycle was analyzed on a flow cytometer using PI / RNase staining solution.
[0058] Experimental results such as figure 2 As shown, 2.5 μM compound 4 can potently induce G2 / M phase arrest in the above-mentioned tumor cells.
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