RRC ID 27188
Author Hung CC, Chiou MH, Teng YN, Hsieh YW, Huang CL, Lane HY.
Title Functional impact of ABCB1 variants on interactions between P-glycoprotein and methadone.
Journal PLoS One
Abstract Methadone is a widely used substitution therapy for opioid addiction. Large inter-individual variability has been observed in methadone maintenance dosages and P-glycoprotein (P-gp) was considered to be one of the major contributors. To investigate the mechanism of P-gp's interaction with methadone, as well as the effect of genetic variants on the interaction, Flp-In™-293 cells stably transfected with various genotypes of human P-gp were established in the present study. The RNA and protein expression levels of human P-gp were confirmed by real-time quantitative RT-PCR and western blot, respectively. Utilizing rhodamine 123 efflux assay and calcein-AM uptake study, methadone was demonstrated to be an inhibitor of wild-type human P-gp via non-competitive kinetic (IC50 = 2.17±0.10 µM), while the variant-type human P-gp, P-gp with 1236T-2677T-3435T genotype and P-gp with 1236T-2677A-3435T genotype, showed less inhibition potency (IC50 = 2.97±0.09 µM and 4.43±1.10 µM, respectively) via uncompetitive kinetics. Methadone also stimulated P-gp ATPase and inhibited verapamil-stimulated P-gp ATPase activity under therapeutic concentrations. These results may provide a possible explanation for higher methadone dosage requirements in patients carrying variant-type of P-gp and revealed the possible drug-drug interactions in patients who receive concomitant drugs which are also P-gp substrates.
Volume 8(3)
Pages e59419
Published 2013-1-1
DOI 10.1371/journal.pone.0059419
PII PONE-D-12-39123
PMID 23527191
PMC PMC3602015
MeSH ATP Binding Cassette Transporter, Subfamily B ATP Binding Cassette Transporter, Subfamily B, Member 1 / genetics ATP Binding Cassette Transporter, Subfamily B, Member 1 / metabolism* Analysis of Variance Blotting, Western Fluoresceins / metabolism Genetic Variation* HEK293 Cells Humans Immunoblotting Inhibitory Concentration 50 Kinetics Methadone / metabolism* Mutagenesis, Site-Directed Real-Time Polymerase Chain Reaction Rhodamine 123 / metabolism Verapamil
IF 2.74
Times Cited 24
DNA material Human MDR1 cDNA (RDB01372)