An Ultra‐Fast UPLC‐MS/MS Method for Quantifying Ivosidenib: Application to Assessment of Metabolic Stability in Human Liver Microsomes

Abstract

Ivosidenib (Tibsovo) is a groundbreaking, first‐in‐class medication that specifically and reversibly blocks isocitrate dehydrogenase (IDH1), earning multiple FDA approvals for cancers harboring IDH1 mutations. This research developed a rapid, dependable, and sensitive UPLC‐MS/MS technique to assess the metabolic stability of ivosidenib (IVB) in human liver microsomes (HLM). The method showed a commendable environmental profile (ComplexMoGAPI score: 69.0, AGREEprep: 0.68, and AGSA: 72.22). Using StarDrop software (including DEREK and P450 tools), chemical structure alerts were detected, and in silico analysis of metabolic vulnerability (CSL value) was conducted. A straightforward isocratic mobile phase system with a stationary phase (C8 column) efficiently separated IVB and baricitinib (BCB as internal standard) in an ultra‐fast 1‐min run at 0.4 ml/min flow rate. The IVB calibration curve showed a linearity over the range 1.0 to 4000 ng/ml. Accuracy and precision during intra‐ and inter‐day assessments in the range of −2.18% to 10.17% and −2.83% to 12.50%, respectively. The current approach uniquely combines HLMs‐based metabolic stability assessment with in silico predictions of metabolic soft spots, toxicity, and ADME, along with dual greenness evaluation, all within a single workflow. The in vitro half‐life (t1/2) of IVB was approximately 44.89 min, with an intrinsic clearance (Clint) of 18.06 ml/min/kg. In silico analysis suggests that IVB derivatives with modifications to the 5‐fluoro‐3‐pyridine group (75% at C21) and the 4‐cyano‐2‐pyridine group (23% at C36) may exhibit varying metabolic stability, warranting further experimental validation.