title: If the KI is defined by the free energy of binding to P-glycoprotein, which kinetic parameters define the IC50 for the Madin-Darby canine kidney II cell line overexpressing human multidrug resistance 1 confluent cell monolayer?
abstract: From previous fits of drug transport kinetics across confluent Madin-Darby canine kidney II cell line overexpressing human multidrug resistance 1 cell monolayers, we found that a drug's binding constant to P-glycoprotein (P-gp) was significantly smaller than its IC(50) when that drug was used as an inhibitor against another P-gp substrate. We tested several IC(50) candidate functions, including the standard function, the Kalvass-Pollack function, and the efflux ratio, to determine whether any of them yielded an IC(50) = K(I), as would be expected for water-soluble enzymes. For the confluent cell monolayer, the IC(50)/K(I) ratio is greater than 1 for all candidate functions tested. From the mass action kinetic model, we have derived a simple approximate equation that shows how the IC(50)/K(I) ratio depends on the elementary rate constants from our mass action model. Thus, the IC(50) will differ between cell lines and tissues, for the same probe substrate and inhibitor, if there are different membrane concentrations of P-gp, or the probe substrate's elementary rate constants, partition coefficient, binding constant to P-gp, passive permeability, and ability to access the other transporters (if any) in the two cell lines. The mass action model and the approximate equation for the IC(50)/K(I) ratio derived here can be used to estimate the elementary rate constants needed to extrapolate in vitro drug-drug interactions for compounds to the in vivo environment.
authors.affiliation: Department of Biology, Drexel University, 32nd and Chestnut Sts., Philadelphia, PA 19104, USA.
authors.forename: Annie Albin
authors.initials: AA
authors.lastname: Lumen
authors.forename: Poulomi
authors.initials: P
authors.lastname: Acharya
authors.forename: Joseph W
authors.initials: JW
authors.lastname: Polli
authors.forename: Andrew
authors.initials: A
authors.lastname: Ayrton
authors.forename: Harma
authors.initials: H
authors.lastname: Ellens
authors.forename: Joe
authors.initials: J
authors.lastname: Bentz
mesh_terms: D018435:ATP Binding Cassette Transporter, Subfamily B; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000187:drug effects; D003198:Computer Simulation; D004077:Digoxin / Q000378:metabolism / Q000493:pharmacokinetics; D004285:Dogs; D004347:Drug Interactions; D019450:Genes, MDR; D006801:Humans; D007668:Kidney / Q000378:metabolism; D008954:Models, Biological; D010599:Pharmacokinetics; D011485:Protein Binding / Q000187:drug effects; D011802:Quinidine / Q000378:metabolism / Q000493:pharmacokinetics; D013816:Thermodynamics
chemical_list: C513055:ABCB1 protein, human; D018435:ATP Binding Cassette Transporter, Subfamily B; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D004077:Digoxin; D011802:Quinidine