Novel anti-tumor compound and preparation method and application thereof
A technology of compounds and tumor inhibitors, applied in the fields of brain tumor inhibitors, glioma inhibitors, and spinal cord tumor inhibitors, which can solve the problems of few compounds and no central nervous system tumor inhibitory activity
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Embodiment 1
[0028] The preparation of embodiment 1 compound I
[0029] A bell of medicinal materials is crushed, impregnated with 95% ethanol for 3 days each time, 5 times in total, the impregnated solutions are combined and then concentrated under reduced pressure to obtain the ethanol extract extract. The ethanol extract was sequentially extracted three times with petroleum ether, concentrated under reduced pressure, and finally obtained the petroleum ether layer extract.
[0030] The petroleum ether layer extract was taken and subjected to preliminary separation by silica gel column chromatography; then purified by semi-preparative HPLC to obtain compound I as a light yellow solid, m.p.208-209°C. ESI-MS m / z:[M+H] + 483. 1 H-NMR (pyridine-d 5 ,400MHz)δ H :2.15(1H,m,H-2a),2.45(1H,m,H-2b),1.35(1H,m,H-3a),1.96(1H,m,H-3b),4.30(1H,d ,J=12.0Hz,H-5),2.85(1H,dd,J=11.6Hz,2.7Hz,H-6),3.56(1H,br d,J=11.9Hz,H-7),2.00(1H ,m,H-8a),1.75(1H,m,H-8b),2.94(1H,m,9a),2.38(1H,br d,J=13.5,H-9b),4.83(1H,s...
Embodiment 2
[0032] Embodiment 2 pharmacological experiment method and preparation
[0033] (1) Cell lines: Human glioblastoma cell lines U87MG and LN229 were purchased from ATCC cell bank in the United States.
[0035] 1) Take out the frozen cells from the liquid nitrogen tank, quickly put them into the preheated 37°C water bath and shake continuously, so that they can be evenly heated and melted within 1 min, and then quickly transfer the thawed frozen cells into the pre-prepared container Centrifuge at low speed for 5 minutes at room temperature (800r / min);
[0036] 2) After centrifugation, discard the upper cryopreservation solution, leaving only the bottom sediment, add a small amount of culture medium, pipette the cells to make them evenly disperse in the culture medium to form a cell suspension, transfer to a 25ml culture bottle and replenish the culture medium to 5ml ;
[0037] 3) Place at 37°C, 5% CO 2 Cultivate in the incubator for 24 hours, discar...
Embodiment 3
[0054] Example 3 In vitro anti-cancer cell proliferation experiment (MTS experiment)
[0055] 1) Cultivate U87MG and LN229 cells until the cells grow to the logarithmic phase under an inverted microscope, digest with trypsin and make a concentration of 1×10 5 cell / ml cell suspension.
[0056] 2) Gently mix the above two cell suspensions, add 100 μl per well into a 96-well plate with a multi-channel micropipette, and the number of cells in each well is 1×10 4 indivual. Each type of cells was seeded into three 96-well plates, and the wells without adding cells, culture medium and drugs were set up as the zero-adjustment group in the experiment. The 96-well plate was cultured overnight in a cell culture incubator until a monolayer of cells was formed covering the bottom of the well.
[0057] 3) In the morning of the next day, after the cell monolayer covered the bottom of the well, the cell culture medium, temozolomide (10nM), temozolomide (100nM), temozolomide (1μM), temozolo...
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