in vitro and in silico evaluation of P-glycoprotein inhibition through 99mTc-MIBI uptake

Hosseini Balef, Seyed Sajad and Piramoon, Majid and Hosseinimehr, Seyed Jalal and Irannejad, Hamid (2019) in vitro and in silico evaluation of P-glycoprotein inhibition through 99mTc-MIBI uptake. Chem Biol Drug Des.

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Abstract

p-glycoprotein (P-gp) is a multi-drug resistance (MDR) transporter with unknown structural details. This macromolecule is normally responsible for extruding xenobiotics from normal cells. Over-expression of P-gp in tumor cells is a major obstacle in cancer chemotherapy. In this article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record. Please cite this article as this study, human 3D model of P-gp was built by homology modeling based on mouse P-gp crystallographic structure and stabilized through 1 ns molecular dynamic (MD) simulation. Stabilized human P-gp structure was used for flexible docking of 80 drugs into the putative active site of P-gp. Accordingly, digoxin, itraconazole, risperidone, ketoconazole, prazosin, verapamil, cyclosporine A, and ranitidine were selected for further in vitro assay. Subsequently, cell based P-gp inhibition assay was performed on Caco-2 cells while 99mTc MIBI was used as a P-gp efflux substrate for calculating IC50 values. Results of the 99mTc MIBI uptake in drugs-treated Caco-2 cells were in agreement with the previously reported activities. This study for the first time described the relation between molecular dynamics and flexible docking with cellular experiments using 99mTc-MIBI radiotracer for evaluation of potencies of P-gp inhibitors. Finally, results showed that our radiotracer-cell based assay is an accurate and fast screening tool for detecting P-gp inhibitors and non-inhibitors in drug development process. Keywords: P-glycoprotein; 99mTc-MIBI; Caco-2; Homology modeling; Flexible docking

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