The purpose of this study was to investigate the effect of probenecid, an inhibitor of the MRP2/ABCC transporter, on the pharmacokinetics and transport of belotecan (7-[2-(N-isopropy-lamino)ethyl]-(20S)-camptothecin). The effect of probenecid on the pharmacokinetics of belotecan was studied in rats. When belotecan was injected as a bolus dose of 5 mg/kg after probenecid was infused at a rate of 42.8 mg/2 mL/h/kg, the cumulative biliary excretion amounts and biliary clearance $(CL_b)$ of belotecan decreased ($28.29{pm}2.83$ versus $19.96{pm}1.45%$ of dose and $161.01{pm}26.95$ versus $92.66{pm}1.45$ mL/min/kg), whereas the systemic pharmacokinetics did not change. This indicates that the MRP2 transporter is involved in the biliary excretion of belotecan. The involvement of MRP2 in the secretory transport was further characterized using Caco-2 cell monolayers expressing MRP2. The apparent permeability across Caco-2 cell monolayers from basolateral to apical was 2.3 times greater than that from the apical to the basolateral side at the $50{mu}M$ belotecan. In addition, probenecid significantly decreased the basolateral-to-apical transport of belotecan (52.9%). These results indicate that MRP2 is involved in the secretory transport of belotecan in biliary excretion.