A rapid and sensitive LC-MS/MS method for evaluation of the absolute oral bioavailability of a novel c-Met tyrosine kinase inhibitor QBH-196 in rats.

Abstract

A sensitive, selective and high-throughput UPLC-MS/MS method was developed and validated for the determination of a novel c-Met tyrosine kinase inhibitor, QBH-196, in rat plasma. QBH-196 and its analog BH357 (IS) were extracted from rat plasma using a mixture of dichloromethane and N-hexane (2:3, v/v). The chromatographic separation was carried out on Phenomenex C18 column (50 × 2.1 mm, 2.6 µm particle size) with a gradient mobile phase of methanol (A) and water containing 0.05% formic acid (B) at a flow rate of 0.2 mL/min. The assay was performed by positive electrospray ionization in multiple reaction monitoring mode using transitions of m/z 622.68 → 140.41 for QBH-196 and m/z 591.19 →126.21 for the IS, respectively. Good linearity was obtained over the concentration range of 8.0-4000 ng/mL (r(2) > 0.99) for QBH-196 and the lower limit of quantification was 8.0 ng/mL in rat plasma. Validations of the method, including its sensitivity, extraction recovery, matrix effect, intra- and inter-day precision, accuracy and stability, were all within acceptable limits. The established method was successfully applied to determine absolute oral bioavailability of QBH-196 in rats for the first time. The mean oral absolute bioavailability of QBH-196 was found to be about 40.8% and the elimination half-life was 40.0 ± 13.1 h. This result suggested that QBH-196 exhibits good oral absorption in vivo, which is very important for the further development of QBH-196 as a new oral anticancer drug.