| Assay Method Information | |
| | URAT1 inhibition assay |
| Description: | The IC50 value of URAT1 inhibition for the crystalline form A of TY706 of the present invention was determined according to the following method:After trypsin digestion, both the expression cells (HEK293) that stably express the URAT1 gene and mock cells were seeded into a lysine-coated 24-well culture plate with a cell seeding density of 1×105 cells/well, and cultured in an incubator with 5% CO2 at 37° C. under saturated humidity for 2 days. The culture medium was removed from the culture plate, and the cultured cells were washed twice with DPBS, and incubated in DPBS buffer at 37° C. for 10 min 500 μL of solutions containing radio-labeled probe substrate ([8-14C] uric acid) and a series of concentrations (0.001-10 μM) of the test compounds or a blank solution were then used to replace DPBS. The concentration of [8-14C] uric acid was 30 μM, and the radiation intensity was 0.867 μCi per well. After 2 min, the reaction was terminated with DPBS buffer solution in ice bath and washed 3 times. Then, 500 μL of 0.1 mol/L NaOH was added to each well to lyse the cells, and the lysate was extracted into a scintillation vial. 3 mL of scintillation liquid (Aquasol-2) was added, and Tri-Carb 2910TR liquid scintillation counter (PerkinElmer, Waltham, USA) was used to determine the radiation intensity in the sample. |
| Affinity data for this assay | |
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