Highly sensitive LC-MRM workflow for quantitation of efflux transporters in rat peripheral blood mononuclear cells: Leveraging ProteoExcelTP with MRM prediction capability

Abstract

Though combinational antiretroviral therapy has been proven highly effective, it suffers from drug-drug interactions, drug resistance and adverse reactions upon their long-term use. Introduction of novel drugs in antiretroviral therapy proposes newer treatment options. However, drug safety and their interaction potential after long-term therapy are still unexplored. In this study, the induction potential of bictegravir on efflux transporters at protein level was assessed here by estimating the transporters by LC-MS/MS based quantitation method. Surrogate peptide approach was used for simultaneous determination of P-gp, BCRP and MRP1 transporter proteins in rat peripheral blood mononuclear cells. Previously developed excel based ProteoExcelTP tool was utilized for selection of surrogate peptides corresponding to target transporters. Moreover, ProteoExcelTP was integrated with a newer MRM prediction capability for prediction of MRM transitions of selected surrogate peptides. Surrogate peptides LLSGQALK (415.2→716.4), SSLLDVLAAR (522.8→288.1) and EDLDLVLK (472.7→685.3) were selected for P-gp, BCRP and MRP1 transporters proteins, respectively. The peptides LLSGQALK, SSLLDVLAAR and EDLDLVLK were eluted at 5.4, 7.0 and 4.1 min, respectively. The finding of the study revealed that bictegravir can significantly induce BCRP transporter after a week of its administration to Sprague Dawley rats. This finding can be utilized in future to prevent transporter mediated drug-drug interactions of bictegravir. Moreover, addition of MRM prediction feature to ProteoExcelTP has enhanced its applicability in mass spectrometry based targeted proteomics. The developed LC-MS/MS based quantitation method for determination of clinically relevant efflux transporters will be useful in investigating the induction potential of any other drug.