CYP450-Mediated metabolism of suvorexant and investigation of metabolites in forensic case specimens

Forensic Sci Int. 2020 Jul;312:110307. doi: 10.1016/j.forsciint.2020.110307. Epub 2020 Apr 28.

Abstract

Suvorexant (Belsomra®) is a sedative hypnotic that was approved for use in 2015. It has a novel mechanism of action and was the first dual orexin receptor antagonist (DORA) to be approved for the treatment of sleep disorders. Sedative hypnotics often feature prominently in forensic investigations such as impaired driving and drug-facilitated sexual assault (DFSA) cases. As such, suvorexant is a drug of interest and its identification in forensic toxicology investigations is of significance. However, limited studies have been published to date and the disposition or importance of its metabolites has been largely uninvestigated. In this report, we investigate the enzymes responsible for metabolism and explore the prevalence of metabolites in blood from a series of thirteen forensic investigations. Recombinant cytochrome P450 enzymes (rCYPs) were used to generate phase I metabolites for suvorexant in vitro, and metabolites were identified using liquid chromatography-quadrupole/time-of-flight-mass spectrometry (LC-Q/TOF-MS). Four rCYP isoenzymes (3A4, 2C19, 2D6, and 2C9) were found to contribute to suvorexant metabolism. The only metabolite identified in blood or plasma arose from hydroxylation of the benzyl triazole moiety (M9). This metabolite was identified in seventeen blood and plasma specimens from twelve medicolegal death investigations and one impaired driving investigation. In the absence of a commercially available reference material, the metabolite was confirmed using rCYP-generated in vitro controls using high resolution mass spectrometry.

Keywords: Blood; CYP450; LC-Q/TOF-MS; Metabolism; Plasma; Suvorexant.

MeSH terms

  • Azepines / chemistry*
  • Azepines / pharmacokinetics*
  • Chromatography, Liquid
  • Cytochrome P-450 Enzyme System / metabolism*
  • Forensic Toxicology / methods
  • Humans
  • Isoenzymes / metabolism
  • Mass Spectrometry
  • Sleep Aids, Pharmaceutical / chemistry*
  • Sleep Aids, Pharmaceutical / pharmacokinetics*
  • Triazoles / chemistry*
  • Triazoles / pharmacokinetics*

Substances

  • Azepines
  • Isoenzymes
  • Sleep Aids, Pharmaceutical
  • Triazoles
  • suvorexant
  • Cytochrome P-450 Enzyme System