ADME Studies-in vitro
All in vitro metabolism studies are performed under GLP and also non-GLP screening is possible. Negative/positive controls and reference substances are included in all in vitro studies. All assays are validated. In vitro ADME studies offered by JOINN include (but are not limited to):
ABSORPTION
Intestinal absorption and Pgp binding：Caco-2 cells/PAMPA, bi-directional transport assay, evaluate intestinal permeability, determine whether a compound is a Pgp substrate or inhibitor
Dermal absorption：animal skin, fresh or frozen, evaluate dermal permeability, mass balance including skin tape stripping
DISTRIBUTION
Plasma protein binding：plasma from different species, equilibrium dialysis, ultrafiltration, SHPLC (large molecular), drug stability in plasma, species comparison
Blood distribution：whole blood of one or more species, determination of the compound between the different blood components(whole blood/ plasma ratio), drug stability in blood
METABOLISM
Cytochrome P450 identification：Identification of specific CYP450 iso-enzymes involved in the metabolism of a drug using a tiered approach with: recombinant enzymes (cDNA expressed iso-enzymes), correlation analysis using human liver microsomes of different donors, specific chemical inhibitors
Cytochrome P450 inhibition：pooled human liver microsomes or cDNA expressed enzymes, CYP isoforms: 1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1 & 3A4, standard substrates (two substrates for CYP3A4 inhibition experiments), IC50 and/or Ki determination, reversible and time dependent inhibition, activity of CYP450 isoforms is measured using LC-MS/MS analysis
Cytochrome P450 induction：fresh human and animal hepatocytes, CYP isoforms: 1A2, 2C9, 2C19 and 3A4, activity of CYP450 isoforms is measured using LC-MS/MS analysis
Metabolite profiling：used for elucidation of metabolic pathway and selection of toxicology species, hepatocytes from different species, determination of drug metabolic stability (in vitro half-life), characterization of the metabolite profile (including structural elucidation) using LC-MS/MS, species comparison