Method for Testing of Drugs Belonging to Substrates and Inhibitors of the Transporter Protein BCRP on Caco-2 Cells

Introduction. Breast cancer resistance protein (BCRP) is an efflux membrane transporter that controls the pharmacokinetics of a large number drugs. Its activity may change when taking some endo- and exogenous substances, thus making it link in drug interactions. Aim. The aim study was to develop method for testing drugs belonging BCRP substrates inhibitors vitro. Materials methods. work performed on Caco-2 cells overexpressing BCRP, cultivation transwell-system consisting apical basolateral chambers. Cells were seeded at bottom chamber, which semipermeable membrane. Primarily, transport substrates: methotrexate, mitoxantrone quercetin evaluated concentration range 1, 5, 10, 50 μM direction from basal chamber one (Papp b-a) opposite a-b). ratio Papp b-a / a-b more than «2» characterizes participation proteins transcellular substances. To confirm their experiment carried out with addition inhibitor, reserpine, medium μM. chambers analyzed by HPLC-MS/MS. Results discussion. methotrexate (1 μM), (1–10 μM) both or transwell-system, content recipient not detected. When became 5 3.38 ± 0.08, indicates involvement transporters its transfer. donor concentrations 10 μM, decreased values below «2». At 2.72 0.16. An increase led 6.18 0.08. With substance indicator but remained above value In microns, "2". Reserpine reduced 3.31 times ( p = 0.0002), elimination asymmetry substance. reserpine 3.36 < 0.0001). results indicate control transfer substances through cellular monolayer. Conclusion. A using (5 (10 as marker (50 inhibitor developed tested cells.