DEVELOPMENT AND VALIDATION OF A HIGH PERFORMANCE LIQUID CHROMATOGRAPHY ASSAY FOR THE DETERMINATION OF A FLUORINATED ANALOGUE OF THALIDOMIDE, N-(2,6-DIOXOPIPERIDIN-3-YL)-3,4,5,6-TETRAFLUOROPHTHALAMIC ACID, AND LENALIDOMIDE

摘要

Fluorinaled derivatives of a hydrolysis metabolite oflhalidomide are being investigated for their anli-cancer potential. In this communication, we report the development and validation of an HPLC assay for the determination of N-(2,6-dioxopiperidin-3-yl)-3,4,5,6-lelrafluorophlhalamic acid (SN 29900) for use in pharmacokinelic measurements of the compound. The inlra- and inter-assay accuracy (98-lO2%) and precision (relative standard deviation <4%) are well within limits of acceptability over the concentration range of 3.12-500.00muM. The lower limit of quantification is 3. 12jxM. Solutions of SN 29900 in dimelhylsulfoxide and plasma were stable during short-term (24 hr) and long-lenn (3 mon) storage, and following three cycles of freezing and thawing. The phar-macokinelics of SN 29900 in C57BI/6 mice were determined after intraperiloneal administration (100 and 500mg/kg; n= 3 mice per time-point). The maximum plasma concentration delected at lOOmg/kgfor SN 29900 was 72.5muM and at 500mg/kg was 1669.9muM. The area under the curve (AUC) at 100 and 500mg/kg was calculated to be 46.1nM.h and 843.8muM.h, respectively. The results indicated that in mice, SN 29900 was cleared faster than lenalidomide, a second generation lhalidomide analogue thai is in clinical use. The maximum plasma concentration of lenalidomide was 337 muM and the AUC, was calculated to be 338 fiM.h following an inlra-periloneal dose at WOmg/kg measured using the same assay validated here for SN29900.