Application of On-Line Enantioselective Capillary Electrophoretic Method: Kinetic and Inhibition Study of CYP3A4 Mediated N-Demethylation of Ketamine.
| Authors | |
|---|---|
| Year of publication | 2014 |
| Type | Conference abstract |
| MU Faculty or unit | |
| Citation | |
| Description | Metabolism of a chiral drug can significantly differ between application of racemate and single enantiomers. Rational drug discovery thus requires an early appraisal of this characteristic impacting on the likely success of a drug candidate in the subsequent clinical testing. Capillary electrophoresis (CE) represents a promising technique to assess the fate of drug enantiomers via in vitro assays. Besides chiral highly effective separations and high throughput, it offers the possibility of implementation of on-line methods where a fused-silica capillary is used not only as a separation column but also as a nanoscale reaction chamber integrating the incubation of an enzymatic reaction, separation of its products and their detection and quantitation into a single fully automated run. After initial attempts, an improved on-line method for determination of enantioselective metabolism of anesthetic ketamine mediated by cytochrome P450 3A4 (CYP3A4) was developed. As mixing of reaction components inside the capillary based on diffusion is generic, robust and rapid, the principle of transverse diffusion of laminar flow profiles was adopted. After the incubation was complete, the reaction products were separated in BGE containing highly sulfated ?-cyclodextrin used as chiral selector. The new approach was thoroughly validated for CYP3A4 mediated N-demethylation pathway of ketamine and applied to the determination of its kinetic parameters and the inhibition characteristics in presence of ketoconazole and dexmedetomidine. The results obtained were in a very good agreement with literature data assessed by different techniques. The final method thus represents a miniaturized and cost-effective tool suitable for screening of stereoselective aspects of drug candidates’ metabolism in early stages of a new drug development. |
| Related projects: |