Application of pharmaceutical composition of fingolimod hydrochloride and curcumenol in preparation of anti-multiple myeloma drug
A technology of fingolimod hydrochloride and multiple myeloma, applied in the field of anticancer drugs, can solve the problems of unseen multiple myeloma, achieve great development prospects and potential application value, induce cell necrosis, and inhibit cell proliferation Effect
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Embodiment 1
[0028] CCK-8 method was used to detect the effects of Fingolimod (FTY720) HCl and Curcumol on the survival rate of MM1S cells alone and in combination.
[0029] Experimental steps:
[0030] (1) Collect MM1S cells in the logarithmic growth phase (purchased from ATCC cell bank, USA), adjust the concentration of the cell suspension, add 100 μL to each well, and plate on a 96-well plate so that the cell density to be tested is 1×10 4 a / mL;
[0031] (2) A zero-adjustment group without adding cells and only culture medium and a DMSO control group with a volume concentration of 0.1% without adding drugs are set, and 3 parallel wells are set for each concentration;
[0032] (3) Add 3 μM Fingolimod (FTY720) HCl, 50 μg / mL Curcumol and 3 μM Fingolimod (FTY720) HCl and 50 μg / mL Curcumol joint preparation respectively, and place in 5% CO by volume 2 , incubate in a cell culture incubator at 37°C for 48 hours;
[0033] (4) Add 10 μL of CCK-8 solution to each well, incubate at 37°C in the...
Embodiment 2
[0036] The synergistic effect of different concentrations of Fingolimod (FTY720) HCl and Curcumol drug combination on MM1S cells was detected by CCK-8 method.
[0037] Experimental steps:
[0038] (1) Collect MM1S cells in the logarithmic growth phase, adjust the concentration of the cell suspension, add 100 μL to each well, and plate on a 96-well plate so that the cell density to be tested is 1×10 4 piece / m L;
[0039] (2) A zero-adjustment group without adding cells and only culture medium and a DMSO control group with a volume concentration of 0.1% without adding drugs are set, and 3 parallel wells are set for each concentration;
[0040] (3) Add different concentrations of Fingolimod (FTY720) HCl and Curcumol preparations, the concentrations of the joint preparations are 2μM+25μg / mL, 2μM+50μg / mL, 2μM+100μg / mL, 3μM+25μg / mL, 3μM+50μg / mL, 3μM+100μg / mL, 4μM+25μg / mL, 4μM+50μg / mL, 4μM+100μg / mL, placed in 5% CO2 by volume, 37°C cell culture incubator for 48 hours;
[0041] (4...
Embodiment 3
[0046] Annexin V 647 / PI method was used to detect the effects of Fingolimod (FTY720) HCl and Curcumol on the apoptosis of MM1S cells alone and in combination.
[0047] Experimental steps:
[0048] (1) Collect the MM1S cells in the logarithmic growth phase, and adjust the concentration of the cell suspension to 2.5×10 5 Each / mL in a 6-well plate, the total volume is 2mL, then add 3μM Fingolimod (FTY720) HCl, 50ug / mL Curcumol, and the combined preparation of 3μM Fingolimod (FTY720) HCl and 50ug / mL Curcumol, and the volume of no drug The DMSO treatment group with a concentration of 0.1% was used as a control, which was placed in a cell culture box and incubated for 27 hours;
[0049] (2) Collect the cells, centrifuge at 2500rpm, 4°C for 5min to collect the cells, wash the cells twice with pre-cooled DPBS, each time at 2500rpm, centrifuge at 4°C for 5min, collect 2-3×10 5 cells;
[0050] (3) Add 100 μL 1×Binding Buffer (Vazyme, Nanjing, China) to resuspend the cells;
[0051] ...
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