OAT、OCT、OATP Transport Substrate Identification And Non-radiolabel Analytical Methodology Investigation

Drug-drug interaction and food-drug interaction have attracted the attention ofinternational academia, because of serious impact on human health. There are manymethods which could be used for studying the interaction. One of the most widelyrecognized methods is human transporter transfected cell lines for in vitro simulation.This paper is founded on studying transporter protein function, non radiolabelmethod establishment, combination of radiolabel and non radiolabel method. Theevaluation model is human transporter transfected cell lines.The main results and conclusions of this paper are as follows. First, the functionof these7transporter cell lines was confirmed. Radiolabeled substrate uptakeenhanced phenomenon of3cell lines was found and illustrated in transport efficiency.Second, LC/MS/MS method for rosuvastatin in cell lysates was established and thismethod was used for studying the function of hOATP1B1, hOATP2B1and hOAT3.This method was also used for determining the affinity parameter Km of hOATP1B1and hOATP2B1. Besides, it was recognied as a new investigation strategy forreplacing the traditional radiolabel quantification method. Third, the affinityrelationship between catalpol and hOCT2was studied and proved. Besides, it wasfound that catalpol probably would be a new promoter of hOCT2because it couldfacilitate C14-TEA absorption. Finally, the inhibitory effect of cinnamic acid, ferulicacid, oleanolic acid, deoxycholic acid and cynarin on hOAT1, hOAT3, hOATP1B1and hOATP2B1was studied for the first time. Cinnamic acid, ferulic acid,deoxycholic acid and cynarin were found and validated as the competitive inhibitorsof hOAT1. Meanwhile, the present study suggested new food-drug interactions.Besides, deoxycholic acid was found as the inhibitor for hOAT1, hOAT3, hOATP1B1and hOATP2B1which probably could be used for revealing the correlation betweenhOATs and hOATPs.