Assessment of Substrate-Dependent Ligand Interactions at the Organic Cation Transporter OCT2 Using Six Model Substrates

Mol Pharmacol. 2018 Sep;94(3):1057-1068. doi: 10.1124/mol.117.111443. Epub 2018 Jun 8.

Abstract

Organic cation transporter (OCT) 2 mediates the entry step for organic cation secretion by renal proximal tubule cells and is a site of unwanted drug-drug interactions (DDIs). But reliance on decision tree-based predictions of DDIs at OCT2 that depend on IC50 values can be suspect because they can be influenced by choice of transported substrate; for example, IC50 values for the inhibition of metformin versus MPP transport can vary by 5- to 10-fold. However, it is not clear whether the substrate dependence of a ligand interaction is common among OCT2 substrates. To address this question, we screened the inhibitory effectiveness of 20 µM concentrations of several hundred compounds against OCT2-mediated uptake of six structurally distinct substrates: MPP, metformin, N,N,N-trimethyl-2-[methyl(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino]ethanaminium (NBD-MTMA), TEA, cimetidine, and 4-4-dimethylaminostyryl-N-methylpyridinium (ASP). Of these, MPP transport was least sensitive to inhibition. IC50 values for 20 structurally diverse compounds confirmed this profile, with IC50 values for MPP averaging 6-fold larger than those for the other substrates. Bayesian machine-learning models of ligand-induced inhibition displayed generally good statistics after cross-validation and external testing. Applying our ASP model to a previously published large-scale screening study for inhibition of OCT2-mediated ASP transport resulted in comparable statistics, with approximately 75% of "active" inhibitors predicted correctly. The differential sensitivity of MPP transport to inhibition suggests that multiple ligands can interact simultaneously with OCT2 and supports the recommendation that MPP not be used as a test substrate for OCT2 screening. Instead, metformin appears to be a comparatively representative OCT2 substrate for both in vitro and in vivo (clinical) use.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • CHO Cells
  • Cimetidine / chemistry
  • Cimetidine / metabolism
  • Cimetidine / pharmacology
  • Cricetinae
  • Cricetulus
  • Dose-Response Relationship, Drug
  • Histamine H2 Antagonists / chemistry
  • Histamine H2 Antagonists / metabolism
  • Histamine H2 Antagonists / pharmacology
  • Hypoglycemic Agents / chemistry
  • Hypoglycemic Agents / metabolism
  • Hypoglycemic Agents / pharmacology
  • Ligands
  • Metformin / chemistry
  • Metformin / metabolism
  • Metformin / pharmacology
  • Models, Chemical*
  • Organic Cation Transporter 2 / agonists
  • Organic Cation Transporter 2 / antagonists & inhibitors
  • Organic Cation Transporter 2 / metabolism*
  • Protein Binding / physiology
  • Substrate Specificity / drug effects
  • Substrate Specificity / physiology

Substances

  • Histamine H2 Antagonists
  • Hypoglycemic Agents
  • Ligands
  • Organic Cation Transporter 2
  • Cimetidine
  • Metformin