Application of the pharmaceutical composition of fingolimod hydrochloride and curcumol in the preparation of anti-multiple myeloma drugs
A technology for fingolimod hydrochloride and multiple myeloma, applied in the field of anti-cancer drugs, can solve the problems of unseen multiple myeloma, achieve great development prospects and potential application value, improve efficiency, and induce cell necrosis.
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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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