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In vitro and in silico evaluation of P‐glycoprotein inhibition through 99m Tc‐methoxyisobutylisonitrile uptake
Author(s) -
Hosseini Balef Seyed Sajad,
Piramoon Majid,
Hosseinimehr Seyed Jalal,
Irannejad Hamid
Publication year - 2019
Publication title -
chemical biology and drug design
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.59
H-Index - 77
eISSN - 1747-0285
pISSN - 1747-0277
DOI - 10.1111/cbdd.13411
Subject(s) - in silico , in vitro , glycoprotein , p glycoprotein , chemistry , computational biology , biochemistry , biology , pharmacology , microbiology and biotechnology , gene , multiple drug resistance , antibiotics
P‐glycoprotein (P‐gp) is a multidrug resistance ( MDR ) transporter with unknown structural details. This macromolecule is normally responsible for extruding xenobiotics from normal cells. Overexpression of P‐gp in tumor cells is a major obstacle in cancer chemotherapy. In 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 dynamics ( 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 99m Tc‐methoxyisobutylisonitrile ( MIBI ) was used as a P‐gp efflux substrate for calculating IC 50 values. Results of the 99m Tc‐ MIBI uptake in drug‐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 99m Tc‐ 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.