A critical assessment on stability behaviour of Vorinostat using LC-MS-QTOF with H/D exchange and NMR

Abstract

Vorinostat is the first USFDA-approved HDAC inhibitor for the treatment of cutaneous t-cell lymphoma. Vorinostat was exposed to ICH-recommended hydrolytic (acid, base, and neutral), oxidative, thermal, and photolytic stress conditions to understand the degradation behaviour. A Stability indicating LC method was developed and validated for separating and identifying forced degradation products. Under different stress conditions, six degradants were identified and characterized by LC-HRMS, MS/MS, and hydrogen-deuterium exchange mass studies. Vorinostat was found to be highly susceptible to the acidic and basic environment. In contrast, the drug substance was stable in the solid state under thermal and photolytic conditions whereas, it was found moderately stable when photolytic stress was provided to dissolved state of Vorinostat in acetonitrile-water. The degradants were identified as 7-amino-N-phenylheptanamide, 8-hydrazineyl-8-oxo-N-phenyloctanamide, 8-oxo-8-(phenylamino)octanoic acid, 8-oxo-8-(2-(7-oxo-7-(phenylamino)heptyl)hydrazineyl)-N-phenyloctanamide, 8,8′-(1-hydroxyhydrazine-1,2-diyl)bis(8-oxo-N-phenyloctanamide), and N1-((8-oxo-8-(phenylamino)octanoyl)oxy)-N8-phenyloctanediamide. The mechanistic explanation for the formation of each degradant in stability conditions has also been derived. The major degradants were also isolated/synthesized and characterized through 1H NMR for preparing impurity standards. Additionally, in-silico toxicity of the degradants was predicted in comparison to the drug, to identify whether any degradant has any specific type of toxicity and requires special focus to set specification limits during formulation development. The predicted toxicity indicated that the degradants have similar safety profile as that of the drug and specification can be set as per general impurity guideline.