Evaluation of Strategies for the Assessment of Drug-Drug Interactions Involving Cytochrome P450 Enzymes

Eur J Drug Metab Pharmacokinet. 2018 Dec;43(6):737-750. doi: 10.1007/s13318-018-0485-7.

Abstract

Background and objectives: Drug-drug interactions (DDIs) can occur when one drug alters the metabolism of another drug. Drug metabolism mediated by cytochrome P450 enzymes (CYPs) is responsible for the majority of metabolism of known drugs and inhibition of CYP enzymes is a well-known cause of DDIs. In the current study, the use of various human liver microsomes (HLM)-based methods to determine occurrence of CYP-mediated metabolism-dependent inhibition (MDI) and possible follow-up studies were evaluated.

Methods: Human CYP inhibition was studied using the following methodologies: direct inhibition and (non-diluted) IC50-shift assays, a ferricyanide-based reversibility assay, a spectrophotometric metabolic intermediate complex (MIC) assay, and recording of reduced carbon monoxide (CO)-difference spectra. HLM incubations in the presence and absence of NADPH and glutathione (GSH) were performed to study the possible formation of CYP-dependent GSH adducts. HLM incubations with the radiolabeled inhibitors mifepristone and paroxetine were performed to study CYP-mediated covalent binding.

Results: Dihydralazine and furafylline displayed irreversible MDI of CYP1A2. Paroxetine displayed both quasi-irreversible and irreversible MDI of CYP2D6, formation of CYP-dependent GSH adducts was observed, while CYP-mediated covalent binding occurred which was decreased in the presence of GSH. Mifepristone displayed irreversible MDI of CYP3A4, formation of CYP-dependent GSH adducts was observed, while CYP-mediated covalent binding occurred which was decreased in the presence of GSH. Troleandomycin and verapamil displayed quasi-irreversible MDI of CYP3A4; MIC formation was observed, while no formation of CYP-dependent GSH adducts occurred.

Conclusions: This study gives a representative overview of current methodologies that can be used to study CYP inhibition. The here presented strategy can be applied as a tool during risk evaluation of CYP-mediated DDIs.

MeSH terms

  • Cytochrome P-450 CYP1A2 Inhibitors / pharmacology*
  • Cytochrome P-450 CYP2D6 Inhibitors / pharmacology*
  • Cytochrome P-450 CYP3A Inhibitors / pharmacology*
  • Dihydralazine / pharmacology
  • Dose-Response Relationship, Drug
  • Drug Interactions
  • Humans
  • Microsomes, Liver / metabolism
  • Mifepristone / metabolism
  • Mifepristone / pharmacology
  • Paroxetine / metabolism
  • Paroxetine / pharmacology
  • Theophylline / analogs & derivatives
  • Theophylline / pharmacology
  • Troleandomycin / pharmacology
  • Verapamil / pharmacology

Substances

  • Cytochrome P-450 CYP1A2 Inhibitors
  • Cytochrome P-450 CYP2D6 Inhibitors
  • Cytochrome P-450 CYP3A Inhibitors
  • Mifepristone
  • Paroxetine
  • Theophylline
  • furafylline
  • Troleandomycin
  • Verapamil
  • Dihydralazine