Quantification of henatinib maleate, a novel potent inhibitor of VEGF receptors, in rat plasma by LC‐MS/MS

Abstract

Henatinib maleate (R,Z)-2-[(5-fluoro-1,2-dihydro-2-oxo-3H-indol-3-ylidene) methyl]-5-(2-hydroxy-3-morpholinopropyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c] azepin-4-ketone maleate is a potent inhibitor of vascular endothelial growth factor receptors, and is currently under preclinical evaluation as an anticancer drug. A novel method for the quantification of henatinib maleate in rat plasma using high performance liquid chromatography-tandem mass spectrometry has been developed. The analyte (henatinib maleate) and internal standard (papaverine hydrochloride) were extracted from 50 microL of rat plasma by protein precipitation and separated on a C(18) column using a mixture of 25 mm ammonium acetate buffer : methanol : acetonitrile (35 : 50 : 15, v/v/v) as mobile phase with a run time of 4.5 min. The detection was performed by means of triple quadrupole mass spectrometer equipped with an ESI interface operating in the multiple-reaction monitoring mode. A linear response was observed over the concentration range 5.0-1000 ng/mL. The limit of quantification was 5.0 ng/mL. Both intra- and inter-day precision, defined as relative standard deviation, were within 9.7%. Accuracy, defined as relative error, was within +/- 3.1%. The developed method was successfully applied to preclinical pharmacokinetic studies of henatinib maleate in rat after a single oral administration of the drug.