Assessment of Apalutamide Metabolic Stability in HLMs Matrix Using a Sensitive and Fast UPLC-MS/MS Method: In Silico Screening for Toxicity Alerts and Metabolic Lability

Apalutamide (Erleada) is an oral Androgen Receptor antagonist that inhibits androgen action, reducing Cancer cell growth. The FDA approved apalutamide (APD) for metastatic castration-sensitive prostate Cancer (mCSPC) and nonmetastatic castration-resistant prostate Cancer. This study created a fast, reliable, and sensitive UPLC-MS/MS method to assess APD metabolic stability in human liver microsomes (HLMs). The greenness of the method was assessed using in silico applications like AGREE and MoGAPI. The StarDrop package (WhichP450 and DEREK modules) identified APD chemical structure warnings and assessed metabolic lability. An isocratic mobile phase system with a C18 column segregated APD and baricitinib (BCB as internal standard). APD calibration curve sensitivity was high, with statistical linearity from 1.0 to 4000 ng/mL. Intraday and interday evaluation accuracy and precision were 0.72%-10.25% and -0.69% to 12.25%, respectively. The UPLC-MS/MS method used a little amount of acetonitrile (35%), a low flow rate of 0.3 mL/min, and a short running period of 1 min, demonstrating environmental sustainability. APD's in vitro half-life (t_1/2) and intrinsic clearance (Cl_int) were 25.57 min and 31.7 mL/min/kg, respectively. These in silico data propose that APD derivatives with changes in N-methyl moiety might reveal altered metabolic lability; this requires experimental verification.

DEREK alerts; StarDrop software; UPLC‐MS/MS; apalutamide; metabolic stability.