p-glycoprotein inhibitor
A P-glycoprotein, active drug technology, applied in the application field of saikosaponin C as a P-glycoprotein inhibitor, can solve the problems of chemotherapy failure, reduce drug efficacy and the like, and achieve the effect of less toxicity
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Embodiment 1
[0015] Study on the effect of saikosaponin C on the uptake of colchicine by normal rat liver cells:
[0016] HEK293 cells were inoculated in six-well plates, and the medium was removed after 2 days, and divided into blank control group, saikosaponin C group and verapamil group; cells in the blank group were not treated, saikosaponin C group was added with 50 μM autumn Narcisine and 100 μg / m saikosaponin C complete medium, while the verapamil group added complete medium containing 50 μM verapamil; the final volume was fixed at 2 ml, and after 48 hours of culture, the PBS buffer The cells were washed once; each group was added with a medium containing 50 μM colchicine, and placed in a 37°C incubator for further incubation for 60 minutes. Finally, collect the cells in a centrifuge tube, discard the drug-containing medium in the well, add pre-cooled PBS to wash once, centrifuge and discard the supernatant; add 200 μl of pure water to each tube of cells, vortex to mix, freeze-thaw ...
Embodiment 2
[0032] Effects of saikosaponin C on the expression of P-glycoprotein in HEK293 cells
[0033] 1. Extraction of total cell protein
[0034] The cells were inoculated in 100×20mm culture dishes, and when the cells reached the logarithmic growth phase, various concentrations of vinegar Bupleurum components were added and incubated for 48 hours; a control group (without drug treatment) was also set up. Then discard the medium in the culture dish, add pre-cooled PBS buffer solution to wash twice, collect the cells in a centrifuge tube; centrifuge, and discard the supernatant. Add 100 μl of RIPA lysate to each dish of cells, pump repeatedly with a 1ml syringe to completely lyse; centrifuge at 4°C and 12,000g for 15 minutes, transfer the supernatant to a new EP tube, and store in a -80°C refrigerator for later use. All operations were performed on ice, and each experiment was repeated 3 times.
[0035] The components and content of the culture medium are as follows:
[0036] DMEM ...
Embodiment 3
[0054] Effects of saikosaponin C on the viability of HEK293 cells
[0055] With 12500cells / cm 2 Density inoculate HEK293 cells and 96-well plates, so that the final volume of each well is 100 μl, absorb the complete medium 2 days after inoculation, add a series of drug-containing medium, and measure the OD value 48 hours after adding the drug by the modified MTT method. Calculate the growth inhibitory rate and half inhibitory concentration IC of the drug on the cells 50 value, the formula is as follows:
[0056] (1) Cell growth inhibition rate IR=1-(OD 570(含药) -OD 570(空白) ) / (OD 570(对照) -OD 570(空白) ), OD 570(含药) and OD 570(对照) are the absorbance values at 570nm in the MTT experiment of the cell samples of drug-added and blank culture medium respectively, OD 570(空白) Absorbance value at 570nm of the culture solution in the zeroing well.
[0057] Since the solubility of saikosaponin C and fatty acids can only reach 1 mg / ml and 100 μg / ml at the maximum; according to saik...
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