A Fast LC-MS/MS Methodology for Estimating Savolitinib in Human Liver Microsomes: Assessment of Metabolic Stability Using In Vitro Metabolic Incubation and In Silico Software Analysis

Abstract

Savolitinib (Orpathys®), was developed by (HUTCHMED (Shanghai, China) and, AstraZeneca (Gaithersburg, Maryland, USA), is an inhibitor of the c-Met receptor tyrosine kinase that is orally bioavailable. It was designed for the treatment of pillary and clear-cell renal-cell carcinoma (RCC), colorectal cancer, gastric cancer, and metastatic non-small-cell lung cancer (NSCLC). The current work aimed to develop a rapid, specific, green, and sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) methodology for estimating savolitinib (SVB) in human liver microsomes (HLMs) with application to an in vitro metabolic stability assessment of SVB in HLMs. The validation steps of the current LC-MS/MS methodology in the HLMs were carried out following US FDA bioanalytical method validation guidelines including sensitivity, selectivity, linearity, accuracy, stability, precision, extraction recovery, and matrix effect. SVB and olmutinib (OLM) were chromatographically separated on an Eclipse Plus C8 column using an isocratic mobile phase. SVB parent ions were generated using the positive mode of an electrospray ionization (ESI) source. SVB daughter ions were detected and quantified using the multiple reaction monitoring (MRM) mode of a triple quadrupole mass analyser. The constructed SVB calibration curve showed linearity over the range from 1 to 3000 ng/mL. The interday and intraday accuracy and precision of the developed LC-MS/MS analytical methodology were −6.67%–4.11% and −0.51%–8.75%, respectively. A lower limit of quantification (LLOQ) of 0.87 ng/mL confirmed the sensitivity of the established method. Furthermore, the eco-scale methodology using the in silico AGREE software was used for the greenness assessment of the current LC-MS/MS method, and the outcomes showed that the established method was very eco-friendly. The intrinsic clearance (Clint) and in vitro half-life (t1/2) of SVB were 33.05 mL/min/kg and 24.54 min, respectively. SVB exhibited a moderate extraction ratio. The current study is the first to establish and validate LC-MS/MS for estimating SVB and assessing the metabolic stability of SVB.