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
|
WOS Category
|
PHARMACOLOGY & PHARMACY
|
Resource |
DNA material |
Human MDR1 cDNA (RDB01372) |