Exact kinetic analysis of passive transport across a polarized confluent MDCK cell monolayer modeled as a single barrier

Tran, Thuy; Mittal, Aditya; Gales, Tracy L.; Maleeff, Beverly E.; Aldinger, Tanya; Polli, Joseph W.; Ayrton, Andrew D.; Ellens, Harma; Bentz, Joe

doi:10.1002/jps.20105

Cited by 66 publications

(87 citation statements)

References 12 publications

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“…For the cell monolayer experiments, digoxin transport was expressed as P app (apparent permeability) and calculated as follows (Tran et al, 2004):…”

Section: Discussionmentioning

confidence: 99%

“…In general, cells were seeded onto permeable filter supports of varying materials and formats, and grown until confluent, polarized monolayers were formed. Some of the laboratories have historical microscopic data to demonstrate formation of cell monolayers (Hidalgo et al, 1989;Tran et al, 2004;Xia et al, 2005). Most laboratories determined the optimal combination of seeding density and time in culture by measuring transepithelial electrical resistance (TEER) and/or paracellular permeability and transport of a P-gp substrate at various seeding densities and times after seeding when they implemented the assay in their company.…”

Section: Methodsmentioning

confidence: 99%

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Variability in P-Glycoprotein Inhibitory Potency (IC₅₀) Using Various in Vitro Experimental Systems: Implications for Universal Digoxin Drug-Drug Interaction Risk Assessment Decision Criteria

et al. 2013

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“…For the cell monolayer experiments, digoxin transport was expressed as P app (apparent permeability) and calculated as follows (Tran et al, 2004):…”

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Variability in P-Glycoprotein Inhibitory Potency (IC₅₀) Using Various in Vitro Experimental Systems: Implications for Universal Digoxin Drug-Drug Interaction Risk Assessment Decision Criteria

et al. 2013

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“…However, most of the interpretations express simplified views and provide approximate solutions for the passive permeability coefficient, P app , by assuming that the system is a single barrier. Even for nonlinear cases, influx or efflux at the apical membrane was simplified by viewing the system as a single barrier and assuming that drug molecules present at either the apical or basolateral compartment have direct access to intracellular enzymes or efflux transporters (Guo et al, 1999;Williams et al, 2002;Troutman and Thakker, 2003b;Tran et al, 2004).…”

mentioning

confidence: 99%

Permeability, Transport, and Metabolism of Solutes in Caco-2 Cell Monolayers: A Theoretical Study

Sun¹,

Pang²

2007

Drug Metab Dispos

102

105

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ABSTRACT:We explored the properties of a catenary model that includes the basolateral (B), apical (A), and cellular compartments via simulations under linear and nonlinear conditions to understand the asymmetric observations arising from transporters, enzymes, and permeability in Caco-2 cells. The efflux ratio (EfR; P app,B3A / P app,A3B ), obtained from the effective permeability from the A3B and B3A direction under linear conditions, was unity for passively permeable drugs whose transport does not involve transporters; the value was unaffected by cellular binding or metabolism, but increased with apical efflux. Metabolism was asymmetric, showing lesser metabolite accrual for the B3A than A3B direction because of inherent differences in the volumes for A and B. Moreover, the net flux (total ؊ passive permeation) due to saturable apical efflux, absorption, or metabolism showed nonconformity to simple The majority of drugs available on the market are in oral dosage forms. For the assessment of permeability and oral drug absorption, in silico models (Stenberg et al., 2001) and high throughput systems, such as the parallel artificial membrane permeability assay (Kansy et al., 1998) and cell-based systems (Hidalgo et al., 1989) exist to relate drug permeability to absorption, especially for compounds that do not undergo intestinal metabolism (Usansky and Sinko, 2005). The most popular high-throughput screening tool for drug permeability is human colon carcinoma (Caco-2) (Hidalgo et al., 1989) or transfected Madin-Darby canine kidney (Irvine et al., 1999) cells. Upon culture, Caco-2 cells differentiate and become confluent to form monolayers with tight junctions and polarized apical/mucosal (A side) and basolateral/serosal (B side) membranes that are structurally and functionally similar to those of enterocytes.ABC efflux transporters such as P-glycoprotein (Pgp) multidrug resistance-associated protein 2 (MRP2), and the breast cancer-resistant protein (BCRP) are expressed on the mucosal membrane of Caco-2 (Hunter et al., 1993;Hirohashi et al., 2000), as are the absorptive transporters, such as the proton-coupled oligopeptide transporter (PEPT1) (Guo et al., 1999) and the organic anion transporting polypeptide (OATP) (Kobayashi et al., 2003). Likewise, on the serosal membrane, basolateral efflux transporters such as MRP3 (Hirohashi et al., 2000) and organic solute transporters ␣ and ␤ (OST␣-OST␤) (Okuwaki et al., 2007) are expressed in Caco-2. In addition, multiple metabolic enzymes such as the sulfotransferases, UDPglucuronosyltransferases (H. Sun, L. Zhang, E. C. Chow, G. Lin, K. S. Pang, Z. Zuo, unpublished data), and the glutathione S-transferases (Peters and Roelofs, 1989) N-(4-[2-(1,2,3,4-tetrahydro-6,7-dimethoxy-2-isoquinolinyl)ethyl]-phenyl)-9,10-dihydro-5-methoxy-9-oxo-4-acridine carboxamide; A, apical; B, basolateral; C D,0 , initial loading concentration in donor side at t ϭ 0; P app , effective permeability; EfR, efflux ratio; MK571,vinyl]phenyl]-(2-dimethylcarbamoylethylsulfanyl)methylsulfanyl] propioni...

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“…Note that the "jump" of the concentration in the cytosol, C C , at the first time point in the simulation (6 min), is not a computational artifact. This is the first time point taken in our experiments (Tran et al, 2004Acharya et al, 2006). This jump is due only to connecting a straight line between t ϭ 0 and t ϭ 6 min and would become smooth if we had plotted earlier time points from the simulation.…”

Section: Lumen Et Almentioning

confidence: 99%

If the K_I 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?

Lumen¹,

Acharya²,

Polli³

et al. 2009

Drug Metab Dispos

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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.

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Exact kinetic analysis of passive transport across a polarized confluent MDCK cell monolayer modeled as a single barrier

Cited by 66 publications

References 12 publications

Variability in P-Glycoprotein Inhibitory Potency (IC₅₀) Using Various in Vitro Experimental Systems: Implications for Universal Digoxin Drug-Drug Interaction Risk Assessment Decision Criteria

Variability in P-Glycoprotein Inhibitory Potency (IC₅₀) Using Various in Vitro Experimental Systems: Implications for Universal Digoxin Drug-Drug Interaction Risk Assessment Decision Criteria

Permeability, Transport, and Metabolism of Solutes in Caco-2 Cell Monolayers: A Theoretical Study

If the K_I 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?

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Exact kinetic analysis of passive transport across a polarized confluent MDCK cell monolayer modeled as a single barrier

Cited by 66 publications

References 12 publications

Variability in P-Glycoprotein Inhibitory Potency (IC50) Using Various in Vitro Experimental Systems: Implications for Universal Digoxin Drug-Drug Interaction Risk Assessment Decision Criteria

Variability in P-Glycoprotein Inhibitory Potency (IC50) Using Various in Vitro Experimental Systems: Implications for Universal Digoxin Drug-Drug Interaction Risk Assessment Decision Criteria

Permeability, Transport, and Metabolism of Solutes in Caco-2 Cell Monolayers: A Theoretical Study

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?

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Variability in P-Glycoprotein Inhibitory Potency (IC₅₀) Using Various in Vitro Experimental Systems: Implications for Universal Digoxin Drug-Drug Interaction Risk Assessment Decision Criteria

Variability in P-Glycoprotein Inhibitory Potency (IC₅₀) Using Various in Vitro Experimental Systems: Implications for Universal Digoxin Drug-Drug Interaction Risk Assessment Decision Criteria

If the K_I 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?