Exploring the structure-activity relationships of ABCC2 modulators using a screening approach

Bioorg Med Chem. 2015 Jul 1;23(13):3513-25. doi: 10.1016/j.bmc.2015.04.029. Epub 2015 Apr 17.

Abstract

ABCC2 is a transporter with key influence on liver and kidney pharmacokinetics. In order to explore the structure-activity relationships of compounds that modulate ABCC2, and by doing so gain insights into drug-drug interactions, we screened a library of 432 compounds for modulators of radiolabeled β-estradiol 17-(β-d-glucuronide) (EG) and fluorescent 5(6)-carboxy-2',7'-dichlorofluorescein transport (CDCF) in membrane vesicles. Following the primary screen at 80μM, dose-response curves were used to investigate in detail 86 compounds, identifying 16 low μM inhibitors and providing data about the structure-activity relationships in four series containing 19, 24, 10, and eight analogues. Measurements with the CDCF probe were consistently more robust than for the EG probe. Only one compound was clearly probe-selective with a 50-fold difference in the IC50s obtained by the two assays. We built 24 classification models using the SVM and fused-XY Kohonen methods, revealing molecular descriptors related to number of rings, solubility and lipophilicity as important to distinguish inhibitors from inactive compounds. This study is to the best of our knowledge the first to provide details about structure-activity relationships in ABCC2 modulation.

Keywords: ABC transporter; ABCC2; CDCF; Efflux; Estradiol glucuronide; Inhibitor; MRP2; Membrane vesicles; Modulator; SVM modelling; Sf9; Stimulator; Structure–activity relationships; Transport; Transport assay.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biological Transport / drug effects
  • Estradiol / analogs & derivatives
  • Estradiol / metabolism
  • Fluoresceins / metabolism
  • Gene Expression
  • High-Throughput Screening Assays
  • Molecular Probes / metabolism
  • Multidrug Resistance-Associated Protein 2
  • Multidrug Resistance-Associated Proteins / agonists*
  • Multidrug Resistance-Associated Proteins / antagonists & inhibitors*
  • Multidrug Resistance-Associated Proteins / genetics
  • Multidrug Resistance-Associated Proteins / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Sf9 Cells
  • Small Molecule Libraries / chemistry
  • Small Molecule Libraries / pharmacology*
  • Spodoptera
  • Structure-Activity Relationship
  • Transport Vesicles / drug effects*
  • Transport Vesicles / metabolism

Substances

  • Fluoresceins
  • Molecular Probes
  • Multidrug Resistance-Associated Protein 2
  • Multidrug Resistance-Associated Proteins
  • Recombinant Proteins
  • Small Molecule Libraries
  • 5(6)-carboxy-2',7'-dichlorofluorescein
  • estradiol-17 beta-glucuronide
  • Estradiol