Overview

Expert study designs, in vitro ADME assays and data interpretation from the in vitro ADME services group at Charles River allow you to characterize your drug candidate's ADME properties and its potential for drug interactions.

Development

In vitro ADME assays are an important part of a successful drug development program aiding in the critical decision making process by offering metabolic information about drug-drug interactions, pharmacokinetics, absorption and potential toxicities. Our wide range of automated in vitro assays can be performed under GLP conditions, if required, yielding information in the areas of metabolism, toxicity and physicochemical characteristics.

Assays

While Charles River has standard procedures for each of these assays listed below, our team of experienced study directors can also assist in designing the best strategy and protocols customized to suit any drug discovery need.

  • CYP450 Inhibition
    • Human liver microsomes or recombinant enzymes
    • Major CYP450 isozymes: 1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 3A4 and 2E1
    • Reversible, metabolic and time-dependent inhibition (IC50 shift)
    • Initial screening assessments (% inhibition) or definitive determinations (IC50, Ki)
    • Kinact/KI determinations
    • Determine formation of MIC and/or covalent binding
    • Activity of CYP450 isoforms measured via LC-MS/MS analysis
    • Drug-drug interaction experiments
  • CYP450 Reaction Phenotyping
    • Human liver microsomes or recombinant enzymes
    • CYP450-selective inhibitors to determine which isozyme(s) are involved in metabolism
    • Metabolite-monitoring option
  • CYP450 Induction
    • Multiple CYP450 isozymes: 1A2, 2B6, 2C9, 2C19 and 3A4
    • Enzyme catalytic activity or mRNA levels
    • Catalytic activity of CYP450 isoforms measured via LC-MS/MS analysis
  • Protein Binding – Rapid Equilibrium Dialysis (RED), Ultrafiltration or Ultracentrifugation (UC)
    • Plasma, serum, whole blood, microsomes, tissue homogenates or purified proteins (albumin, AGP)
    • Rodent and non-rodent species upon request
    • Initial screening assessments or definitive binding determinations
    • Ex vivo samples (plasma, tissue homogenates)
  • Melanin Binding
    • Separate free compound from melanin-bound by centrifugation
    • Synthetic melanin
    • Also available from sepia (cuttlefish)
    • Initial screening assessments (% bound) or definitive binding determinations (kd, Bmax)
  • Metabolic Stability – Microsomes, S9
    • Liver microsomes, S9
    • Rodent and non-rodent species upon request
    • Initial screening assessments (% remaining) or definitive determinations (T1/2, CLint)
    • Metabolite-monitoring option
  • Metabolic Stability – Hepatocytes
    • Pooled cryopreserved hepatocytes
    • Fresh primary hepatocytes available upon request
    • Rodent and non-rodent species upon request
    • Initial screening assessments (% remaining) or definitive determinations (T1/2, CLint)
    • Metabolic monitoring option
  • Metabolite ID/Profiling
    • Liver microsomes, S9, hepatocytes, ex vivo samples, plant or sediment material
    • Rodent and non-rodent species upon request
    • Initial screening assessments (profiling) or more definitive determinations (ID) using accurate mass
  • Matrix Stability (Matrix, Buffer)
    • Ex vivo matrices (plasma, serum, blood, tissue, homogenates, CSF), aqueous buffers
    • Initial screening assessments (% remaining) or definitive determinations (T1/2)
  • Red Blood Cell (RBC) Partitioning (Blood/Plasma Ratio)
    • Fresh whole blood
    • Rodent and non-rodent species upon request
    • Blood/plasma ratio, Kp
  • Permeability and Efflux (Caco-2, MDCK, MDCK-MDRI, MDCK-BCRP)
    • Caco-2 (human epithelial colorectal adenocarcinoma cell line), MDCK (wild type, canine), MDCK-MDRI (transfected with human Pgp) and MDCK-BCRP (transfected with human breast cancer resistance protein)
      • Cell monolayers available every week
    • Uni-directional (A-B flux) or bi-directional (efflux)
    • Measurement of analyte levels on apical and basolateral sides of cell monolayer using LC-MS/MS bioanalysis
  • Influx Transporters
    • TransportoCells (HEK 293) from Corning over expressing: OATP1B1, OATP1B3, OAT1, OAT3, OCT1 or OCT2
    • Substrate and inhibition assay formats
    • Initial screening assessments or definitive determinations
  • hERG Inhibition

    hERG Inhibition

  • Cell Proliferation and Cytoxicity

    Cell Proliferation and Cytoxicity