Development and validation of LC-MS/MS method for estimating the pharmacokinetics, protein binding, and metabolic stability of soluble epoxide hydrolase inhibitor EC5026

EC5026 is a soluble Epoxide Hydrolase (SEH) inhibitor which is being developed clinically (Phase-1) as a first-in-class analgesic for the treatment of pain. In the present study, we report the development and validation of the LC-MS/MS method for the estimation of EC5026 from rat plasma. The developed method is simple, specific, and sensitive for the quantification of EC5026 from rat plasma. The method was applied to investigate in-vivo pharmacokinetics after single oral administration (P.O.) of EC5026 (5.0 mg/kg) in Wistar rats. Further, the in-vitro metabolic stability and rat plasma protein binding of EC5026 were evaluated using the developed method. The in-vitro results demonstrated a moderate clearance with a value of 10.8 mL/min/kg and half-life (t1/2) of 57.75 min. The results show moderate clearance by the liver manifesting in satisfactory oral bioavailability. The rat plasma protein binding was estimated to be 96.24% ± 0.97% and 96.38% ± 0.56% at 1 µM and 10 µM concentrations, respectively. The developed analytical method is expected to facilitate future pre-clinical, clinical investigations of EC5026.

Bioanalysis; EC5026; LCMS/MS; Metabolic stability; Pharmacokinetics; Protein Binding.