Screening of OATP1B1/3 and OCT1 inhibitors in cryopreserved hepatocytes in suspension

Badolo, Lassina; Rasmussen, L. M.; Hansen, Hanna E.; Sveigaard, Christina

doi:10.1016/j.ejps.2010.03.023

Cited by 33 publications

(34 citation statements)

References 16 publications

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“…These findings are consistent with previous results from Chang et al (2005), who constructed a pharmacophore model for OATP1B1 and concluded that lipophilicity and hydrogen bond-acceptor strength are important properties for OATP1B1 inhibition. Our results also confirm the conclusions drawn by Badolo et al (2010), who determined that lipophilicity and the number of hydrogen bond acceptors (next to polarity and pKa) are the key properties for inhibiting the OATP1B1 probe substrate estradiol-17b-glucuronide in human hepatocytes. The key descriptors identified with our computational model are consistent with the data published by Karlgren et al (2012b), who concluded that lipophilicity and polar surface area are key molecular features for inhibition of several OATP1B isoforms.…”

Section: Discussionsupporting

confidence: 91%

“…In this respect, the experimental conditions used in our in vitro model differ from previously reported methodologies using an excess of inhibitor. Applying a high inhibitor/substrate concentration ratio might indeed lead also to the selection of those compounds with a relatively lower OATP inhibition potency (Badolo et al, 2010;Karlgren et al, 2012a). In the present study, rifampicin and darunavir inhibited sodium fluorescein uptake by more than 80 and 60% of the control value (Fig.…”

Section: Discussionmentioning

confidence: 61%

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Structure-Based Identification of OATP1B1/3 Inhibitors

et al. 2013

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Several recent studies show that inhibition of the hepatic transport proteins organic anion-transporting polypeptide 1B1 (OATP1B1) and 1B3 (OATP1B3) can result in clinically relevant drug-drug interactions (DDI). To avoid late-stage development drug failures due to OATP1B-mediated DDI, predictive in vitro and in silico methods should be implemented at an early stage of the drug candidate evaluation process. In the present study, we first developed a high-throughput in vitro transporter inhibition assay for the OATP1B subfamily. A total of 2000 compounds were tested as potential modulators of the uptake of the OATP1B substrate sodium fluorescein, in OATP1B1-or 1B3-transfected Chinese hamster ovary cells. At an equimolar substrate-inhibitor concentration of 10 mM, 212 and 139 molecules were identified as OATP1B1 and OATP1B3 inhibitors, respectively (minimum 50% inhibition). For 69 compounds, previously not identified as OATP1B inhibitors, concentrationdependent inhibition was also determined, yielding K i values ranging from 0.06 to 6.5 mM. Based on these in vitro data, we subsequently developed a proteochemometrics-based in silico model, which predicted OATP1B inhibitors in the test group (20% of the dataset) with high specificity (86%) and sensitivity (78%). Moreover, several physicochemical compound properties and substructures related to OATP1B1/1B3 inhibition or inactivity were identified. Finally, model performance was prospectively verified with a set of 54 compounds not included in the original dataset. This validation indicated that 80 and 74% of the compounds were correctly classified for OATP1B1 and OATP1B3 inhibition, respectively.

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Section: Discussionsupporting

confidence: 91%

Section: Discussionmentioning

confidence: 61%

Structure-Based Identification of OATP1B1/3 Inhibitors

et al. 2013

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“…These key interactions are further substantiated by Gui et al (2009) who by use of a comparative molecular field analysis approach identified hydrophobicity and basicity of the side chain residues as being critical for OATP1B1 inhibition. Badolo et al (2010) investigated the ability of 179 compounds to inhibit the uptake of estradiol-17␤-glucuronide in human hepatocytes to develop a structure-activity relationship for OATP1B1/1B3. Lipophilicity, polarity, pK a , and the number of hydrogen bond donors and acceptors again were shown to play a critical role in determining the molecular interactions with the OATP1B1/1B3.…”

Section: Discussionmentioning

confidence: 99%

The Development, Characterization, and Application of an OATP1B1 Inhibition Assay in Drug Discovery

Soars¹,

Barton²,

Ismair³

et al. 2012

Drug Metab Dispos

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ABSTRACT:The pivotal role of organic anion-transporting polypeptide 1B1 (OATP1B1) in drug disposition has become clear over the last decade. Therefore, an OATP1B1 inhibition assay suitable for use within early drug discovery was developed and characterized. IC 50 estimates for 10 literature compounds using pitavastatin and estradiol-17␤-glucuronide as substrates were within 2-fold of each other. In addition, the IC 50 estimates using pitavastatin uptake agreed well with literature values (r 2 ‫؍‬ 0.92, average fold error ‫؍‬ 1.3).However, when estrone-3-sulfate was used, OATP1B1 inhibition was underpredicted by as much as 10-fold. A comparison of uptake in human hepatocytes and OATP1B1 inhibition showed a significant correlation (r 2 ‫؍‬ 0.53, P < 0.001) for more than 40 compounds. These data suggest that, for discrete chemical series, OATP1B1 inhibition data may be used as a surrogate for more costly and time-consuming uptake studies in hepatocytes. OATP1B1 inhibition data, determined for over 260 compounds representing both internal AstraZeneca and literature chemistry, were also used to generate a continuous in silico model. The robustness of the model was demonstrated by accurately predicting OATP1B1 inhibition for external test sets using 50 AstraZeneca compounds (root mean square error ‫؍‬ 0.45) and 12 literature drugs (RMSE ‫؍‬ 0.32). The most important molecular descriptors for the prediction of OATP1B1 inhibition were maximal hydrogen bonding strength followed by cLogP. This study has shown that a well validated OATP1B1 inhibition assay in conjunction with an in silico approaches has the potential to influence significantly the designmake-test cycle and subsequently reduce the propensity of OATP1B1 ligands.

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“…Total OATP hepatic uptake is conserved between rodents and humans at a gross kinetic level, with generally good translation of overall apparent substrate or inhibitor affinity and collective impact of OATPs on pharmacokinetics (Badolo et al, 2010;Iusuf et al, 2012b). dx.doi.org/10.1124/dmd.113.054783. s This article has supplemental material available at dmd.aspetjournals.org.…”

Section: Introductionmentioning

confidence: 99%

Utility of Oatp1a/1b-Knockout and OATP1B1/3-Humanized Mice in the Study of OATP-Mediated Pharmacokinetics and Tissue Distribution: Case Studies with Pravastatin, Atorvastatin, Simvastatin, and Carboxydichlorofluorescein

Higgins

Bao

et al. 2013

Drug Metab Dispos

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Although organic anion transporting polypeptide (OATP)-mediated hepatic uptake is generally conserved between rodents and humans at a gross pharmacokinetic level, the presence of three major hepatic OATPs with broad overlap in substrate and inhibitor affinity, and absence of rodent-human orthologs preclude clinical translation of single-gene knockout/knockin findings. At present, changes in pharmacokinetics and tissue distribution of pravastatin, atorvastatin, simvastatin, and carboxydichlorofluorescein were studied in oatp1a/1b-knockout mice lacking the three major hepatic oatp isoforms, and in knockout mice with liver-specific knockin of human OATP1B1 or OATP1B3. Relative to wild-type controls, oatp1a/1b-knockout mice exhibited 1.6-to 19-fold increased intravenous and 2.1-to 115-fold increased oral drug exposure, due to 33%-75% decreased clearance, 14%-60% decreased volume of distribution, and £74-fold increased oral bioavailability, with the magnitude of change depending on the contribution of oatp1a/1b to pharmacokinetics. Hepatic drug distribution was 4.2-to 196-fold lower in oatp1a/1b-knockout mice; distributional attenuation was less notable in kidney, brain, cardiac, and skeletal muscle. Knockin of OATP1B1 or OATP1B3 partially restored control clearance, volume, and bioavailability values (24%-142% increase, £47% increase, and £77% decrease vs. knockout, respectively), such that knockin pharmacokinetic profiles were positioned between knockout and wild-type mice. Consistent with liver-specific humanization, only hepatic drug distribution was partially restored (1.3-to 6.5-fold increase vs. knockout). Exposure and liver distribution changes in OATP1B1-humanized versus knockout mice predicted the clinical impact of OATP1B1 on oral exposure and contribution to human hepatic uptake of statins within 1.7-fold, but only after correcting for human/humanized mouse liver relative protein expression factor (OATP1B1 = 2.2, OATP1B3 = 0.30).

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Screening of OATP1B1/3 and OCT1 inhibitors in cryopreserved hepatocytes in suspension

Cited by 33 publications

References 16 publications

Structure-Based Identification of OATP1B1/3 Inhibitors

Structure-Based Identification of OATP1B1/3 Inhibitors

The Development, Characterization, and Application of an OATP1B1 Inhibition Assay in Drug Discovery

Utility of Oatp1a/1b-Knockout and OATP1B1/3-Humanized Mice in the Study of OATP-Mediated Pharmacokinetics and Tissue Distribution: Case Studies with Pravastatin, Atorvastatin, Simvastatin, and Carboxydichlorofluorescein

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