To confirm the assumption that 6-hydroxy-5, 7-dimethyl-2-methylamino-4-(3-pyridylmethyl)benzothiazole (E3040) acts on urate reabsorption by inhibiting urate-anion exchange at the luminal membrane of renal tubules, we investigated the inhibitory effect of E3040 and its two conjugated metabolites on hydroxyl ion (OH(-)) gradient-dependent urate uptake into brush border membrane vesicles from rat renal cortex and compared it with other uricosuric agents. The order of potency was AA193 (5-chloro-7, 8-dihydro-3-phenylfuro[2,3-g]-1,2-benzisoxazole-7-carboxylic acid)>benzbromarone>E3040>probenecid>E3040 sulfate>E3040 glucuronide. Furthermore, kinetic analysis revealed that E3040 may be a competitive inhibitor of the OH(-) gradient-dependent uptake of urate into brush border membrane vesicles.