Identification of natural products as modulators of OATP2B1 using LC-MS/MS to quantify OATP-mediated uptake

Organic anion-transporting polypeptide 2B1 (OATP2B1) which is highly expressed in enterocytes and hepatocytes could be a key determinant for the intestinal absorption and hepatic uptake of its substrate drugs. Natural products are commonly used in traditional Chinese medicine, foods, and beverages.

The objective of this study is to determine the OATP2B1-mediated drug interactions that could occur between natural products and OATP2B1 substrate drugs.

Human OATP2B1 was transiently expressed in human embryonic kidney (HEK293) cells and characterized by immunofluorescence, Western blot, and uptake assay. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods for detecting OATP2B1 substrates estrone-3-sulfate (E3S) and three statins had been developed and were employed to investigate the effects of 27 frequently used natural products on the function of OATP2B1. Uptake of 5 μM E3S and 1 μM statins in the absence or presence of natural products was measured at 37 °C for 2 min with empty vector- and OATP2B1-transfected HEK293 cells. The IC50 values of inhibitors for OATP2B1-mediated 5 μM E3S uptake were determined.

Our results showed that mulberrin, scutellarin, quercetin, and glycyrrhetinic acid were strong inhibitors of OATP2B1-mediate E3S uptake with IC50 values being 1.8, 2.0, 7.5, and 13.0 μM, which were comparable with their plasma concentrations in clinical trials. They also inhibited OATP-mediated uptake of atorvastatin, fluvastatin, and rosuvastatin. These results indicated that clinically relevant drug interactions could occur between these natural compounds and OATP2B1 substrate drugs.

The information obtained from this study might be helpful to predict and to avoid potential OATP2B1-mediated drug interactions.