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