Pharmaceutical composition containing MLN4924 and Sorafenib and application thereof
A composition and drug technology, applied in the direction of drug combination, medical preparations containing active ingredients, active ingredients of heterocyclic compounds, etc., can solve the problem of MLN4924 liver cancer and achieve good application prospects
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Embodiment 1
[0030] Embodiment 1HepG2 and LO2 cell culture
[0031] 1. HepG2 cell culture
[0033] a. Preparation before resuscitating cells: DMEM cell culture medium preparation and preheating;
[0034] b. Take cells: transfer the frozen cell suspension to a centrifuge tube containing DMEM medium;
[0035] c. Centrifuge, resuspend, and transfer to a culture bottle for culture, while observing the cell growth. If the cell density is too high, passage in time for subsequent experiments.
[0036] 2) Subculture of cells
[0037] a. Observe cell growth: Take out the cells from the incubator, put them under an inverted microscope to observe the cell shape and the number of cells, and observe whether the cells need to be subcultured and the dilution factor required for cell subculture; then put them back into the incubator.
[0038] b. Take cells: take out the cells from the incubator, unscrew the bottle cap after passing the flame of the alcohol lamp on the bottle...
Embodiment 2
[0049] Example 2 The effect of the combination of MLN4924 and Sora on the proliferation of HepG2 cells
[0050] The substance mass ratio range of MLN4924 and Sora of the present invention is 1:160 to 16:5, as shown in Table 1, under the same volume, 0.025 μmol / L MLN4924 combined with 4 μmol / L Sora medication, the ratio range is 1:160; Next, 0.8μmol / LMLN4924 combined with 0.25μmol / L Sora, the ratio range is 16:5.
[0051] Under the same volume, the MLN4924 is 0.025μmol / L, 0.05μmol / L, 0.1μmol / L, 0.2μmol / L, 0.4μmol / L, 0.8μmol / L according to the concentration; the Sora is 0.25μmol / L, 0.5 μmol / L, 1μmol / L, 2μmol / L, 4μmol / L; the concentration and ratio of MLN4924 combined with Sora are as shown in Table 1, and the combined medication is carried out.
[0052] Take the HepG2 in the logarithmic phase of Example 1, digest with trypsin, count the cells, and place them in a 96-well plate at a rate of 2×10 per well. 3 Add 100 μL of cell suspension to inoculate each cell, and store at 37°C...
Embodiment 3
[0074] Example 3 Effects of different doses of MLN4924 and Sora combined on the growth of LO2 cells
[0075] The concentration of MLN4924 is 0.025μmol / L, 0.05μmol / L, 0.1μmol / L, 0.2μmol / L, 0.4μmol / L, 0.8μmol / L; the concentration of Sora is 0.25μmol / L, 0.5μmol / L, 1 μmol / L, 2 μmol / L, 4 μmol / L; MLN4924 combined with Sora, the concentration and ratio of the drug are shown in Table 1, and the combined drug is used.
[0076] The LO2 cells in the logarithmic phase were digested with trypsin, counted, and placed in 96-well plates at 2×10 per well. 3 Each cell was inoculated by adding 100 μL of cell suspension, and the surrounding wells were filled with sterile PBS. Cells were placed in an incubator at 37°C, 5% CO 2 Cultivate until the cells are about 70% confluent; discard the DMEM medium, add 200 μL of medium containing different drug concentrations prepared according to the above ratio and continue to culture for 72 hours, add 10 μL of CCK8 solution to each well, and continue to in...
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