Biotransformation of Daclatasvir In Vitro and in Nonclinical Species: Formation of the Main Metabolite by Pyrrolidine δ-Oxidation and Rearrangement

Drug Metab Dispos. 2016 Jun;44(6):809-20. doi: 10.1124/dmd.115.068866. Epub 2016 Mar 30.

Abstract

Daclatasvir is a first-in-class, potent, and selective inhibitor of the hepatitis C virus nonstructural protein 5A replication complex. In support of nonclinical studies during discovery and exploratory development, liquid chromatography-tandem mass spectrometry and nuclear magnetic resonance were used in connection with synthetic and radiosynthetic approaches to investigate the biotransformation of daclatasvir in vitro and in cynomolgus monkeys, dogs, mice, and rats. The results of these studies indicated that disposition of daclatasvir was accomplished mainly by the release of unchanged daclatasvir into bile and feces and, secondarily, by oxidative metabolism. Cytochrome P450s were the main enzymes involved in the metabolism of daclatasvir. Oxidative pathways included δ-oxidation of the pyrrolidine moiety, resulting in ring opening to an aminoaldehyde intermediate followed by an intramolecular reaction between the aldehyde and the proximal imidazole nitrogen atom. Despite robust formation of the resulting metabolite in multiple systems, rates of covalent binding to protein associated with metabolism of daclatasvir were modest (55.2-67.8 pmol/mg/h) in nicotinamide adenine dinucleotide phosphate (reduced form)-supplemented liver microsomes (human, monkey, rat), suggesting that intramolecular rearrangement was favored over intermolecular binding in the formation of this metabolite. This biotransformation profile supported the continued development of daclatasvir, which is now marketed for the treatment of chronic hepatitis C virus infection.

MeSH terms

  • Animals
  • Bile / metabolism
  • Biotransformation / physiology*
  • Carbamates
  • Chromatography, High Pressure Liquid / methods
  • Cytochrome P-450 Enzyme System / metabolism
  • Dogs
  • Haplorhini
  • Hepatocytes / metabolism
  • Humans
  • Imidazoles / metabolism*
  • Macaca fascicularis
  • Magnetic Resonance Spectroscopy / methods
  • Male
  • Mass Spectrometry / methods
  • Mice
  • Mice, Inbred BALB C
  • Microsomes, Liver / metabolism
  • Oxidation-Reduction
  • Pyrrolidines / metabolism*
  • Rats
  • Rats, Sprague-Dawley
  • Valine / analogs & derivatives

Substances

  • Carbamates
  • Imidazoles
  • Pyrrolidines
  • imidazole
  • Cytochrome P-450 Enzyme System
  • Valine
  • daclatasvir
  • pyrrolidine