Monitoring Cyp2b10 mRNA expression at cessation of 2-year carcinogenesis bioassay in mouse liver provides evidence for a carcinogenic mechanism devoid of human relevance: the dalcetrapib experience

Toxicol Appl Pharmacol. 2012 Mar 15;259(3):355-65. doi: 10.1016/j.taap.2012.01.014. Epub 2012 Jan 28.

Abstract

Introduction: Dalcetrapib is a cholesteryl ester transfer protein (CETP) modulator in clinical assessment for cardiovascular outcome benefits. In compliance with regulatory requirements, dalcetrapib was evaluated in rodent 2-year carcinogenesis bioassays. In the mouse bioassay, male mice demonstrated increased liver weight and statistically increased incidences of hepatocellular adenoma/carcinoma. Hepatic cytochrome p450 (Cyp) 2b10 mRNA induction and increased Cyp2b10 enzyme activity signify activation of hepatic nuclear receptor constitutive androstane receptor (CAR), a widely established promoter of rodent-specific hepatic tumors. We therefore monitored hepatic Cyp2b10 mRNA and its enzyme activity in a subset of dalcetrapib-treated male mice from the bioassay.

Methods: Liver samples were obtained from ~1/3 of male mice from each dose group including vehicle-controls (mean and earliest study day of death 678 and 459 respectively). Quantitative real time PCR (qRT-PCR) was performed to determine Cyp2b10 mRNA expression and Cyp1a-, Cyp2b10- and Cyp3a-selective activities were monitored.

Results: Cyp2b10 mRNA was strongly induced by dalcetrapib with an expected wide inter-individual variation (5-1421-fold). Group average fold-induction versus vehicle-controls showed a dose-related increase from 48-fold (250mg/kg/day) to 160-fold (750mg/kg/day), which declined slightly at 2000mg/kg/day (97-fold). Cyp enzyme activities showed approximate doubling of total Cyp P450 content per milligram protein and a 9-fold increase in Cyp2b10-selective pentoxyresorufin O-dealkylase activity (750mg/kg/day).

Discussion: These data from hepatic Cyp2b10 monitoring are strongly suggestive of CAR activation by dalcetrapib, a mechanism devoid of relevance towards hepatocarcinogenesis in humans; results show feasibility of Cyp2b10 as a surrogate marker for this mechanism at cessation of a carcinogenesis bioassay.

MeSH terms

  • Amides
  • Animals
  • Anticholesteremic Agents / administration & dosage
  • Anticholesteremic Agents / toxicity*
  • Aryl Hydrocarbon Hydroxylases / genetics*
  • Biological Assay / methods
  • Cytochrome P450 Family 2
  • Dose-Response Relationship, Drug
  • Drug Monitoring / methods
  • Esters
  • Feasibility Studies
  • Gene Expression Regulation, Enzymologic / drug effects
  • Humans
  • Liver / drug effects*
  • Liver / enzymology
  • Liver / metabolism
  • Male
  • Mice
  • Mice, Inbred ICR
  • Polymerase Chain Reaction
  • RNA, Messenger / metabolism*
  • Species Specificity
  • Steroid Hydroxylases / genetics*
  • Sulfhydryl Compounds / administration & dosage
  • Sulfhydryl Compounds / toxicity*
  • Time Factors

Substances

  • Amides
  • Anticholesteremic Agents
  • Esters
  • RNA, Messenger
  • Sulfhydryl Compounds
  • dalcetrapib
  • Steroid Hydroxylases
  • Aryl Hydrocarbon Hydroxylases
  • Cyp2b10 protein, mouse
  • Cytochrome P450 Family 2