Aim: Screening of in vitro metabolic stability of test item in plasma from human and preclinical species.
Background information: In addition to hepatic metabolism, drugs can be metabolized by enzymes present in plasma, such as esterases. Structures containing amide and ester groups are susceptible to hydrolysis in plasma.
Drugs with rapid hydrolysis in plasma tend to have high clearance, which leads generally to poor in vivo efficacy. In case of prodrugs, plasma stability may be critical to account for their activity. In addition, plasma stability may show interspecies differences.
Plasma stability is assessed after incubation at preset time points. After stopping the reaction and centrifugation, compound is determined in the supernantant by UPLC-MS/MS.
Compound requirement: 50 µl of 10 mM DMSO solution
Turnaround time: 2 weeks / 5 compounds / 3 species
Test compound concentration: 5 µM (variable on request)
Number of replicates: 2
DMSO concentration: 2%
Incubation time: 0 and 60 min (variable on request)
Control: positive control with known stability
Analytical method: UPLC-MS/MS
% test compound remaining at each time point relative to time 0