Green Ultra-Fast UPLC-MS/MS Method for Enasidenib Quantification in HLMs Matrix With the Metabolic Stability...

Area of Science:

• Analytical Chemistry
• Pharmacology
• Drug Metabolism

Background:

• Enasidenib (ESD) is an oral medication for refractory or relapsed Acute Myeloid Leukemia.
• No prior UPLC-MS/MS method combined green metrics and in silico metabolism analysis for ESD.

Purpose of the Study:

• Establish an ultrafast UPLC-MS/MS method for estimating Enasidenib in Human Liver Microsomes (HLMs).
• Evaluate the greenness and in silico metabolism of Enasidenib.
• Determine the in vitro metabolic stability of Enasidenib.

Main Methods:

• Ultrafast UPLC-MS/MS analysis using positive ESI ionization and a C8 reversed-phase column within one minute.
• Assessment of method greenness using MoGAPI and AGREE scores.
• In vitro metabolic stability assays (t1/2, Clint) and in silico analysis.

Main Results:

• The UPLC-MS/MS method demonstrated good greenness (MoGAPI score 68.0, AGREE score 0.66).
• The method showed linearity from 1 ng/mL to 4000 ng/mL with acceptable precision and accuracy.
• Enasidenib exhibited a long in vitro half-life (82.52 min) and low intrinsic clearance (9.83 mL/min/kg), classifying it as a low-clearance drug.
• In silico analysis suggested potential for enhanced metabolic stability via structural modifications.

Conclusions:

• The developed ultrafast UPLC-MS/MS method is suitable for Enasidenib estimation and metabolic screening.
• Enasidenib is a low-clearance drug, and its metabolic stability can be potentially improved through targeted drug design.
• The combined in vitro/in silico approach offers an efficient strategy for early-stage drug metabolism studies.