Forced Degradation Profiling and Putative Structural Characterization of Atogepant Degradation Products by LC‐QTOF‐MS/MS Supported by In‐Silico Degradation‐Pathway and Toxicity Assessments
Jayaram Kamma, Murugesan Palanivelu, Venkata Narayana Bypaneni, Saikrishna Bompelliwar, Prabaharan Ramachandran, Padmakar Gandla, Anil Kumar RamgiriABSTRACT
Atogepant is an approved medicine prescribed to be administered by oral route to help prevent migraine headaches in adults. The present study aimed to develop a stability‐indicating RP‐HPLC method for quantifying atogepant and its degradation products (DPs) in atogepant active ingredient. The structure assessment of its DPs was conducted using quadrupole time‐of‐flight mass spectrometry (QTOF‐MS). Stress tests of atogepant demonstrated three DPs, one from alkaline hydrolysis (m/z 603) and two from peroxide oxidation (m/z 619, m/z 619). Under alkali stress, atogepant degraded 1.9%, and under oxidative conditions, 4.9%. In this investigation, the developed RP‐HPLC approach employed a Phenomenex Kinetex C18 column and an acetonitrile/trifluoroacetic acid in water eluent. The method was fully validated, demonstrating good precision, linearity, accuracy, and specificity. The calibration curve was observed to be highly linear for the assay of atogepant in the range of 0.25–0.75 mg/mL and for the determination of atogepant DPs in the range of 0.25–1.5 µg/mL. This approach is highly sensitive and allows for the determination of atogepant and their DPs as low as 0.05% of test concentration. The degradation pathway score of the detected degradation components was computed using Zeneth software. The observed impurities were predicted for toxicology by tox‐prediction software, and it was found that one oxidation degradation product is under toxicity class 5 and the other oxidation degradation product and alkaline hydrolysis product are under class 4, suggesting a comparable safety profile with atogepant. In conclusion, the degradation profiling and established stability‐indicating method provide important insights into the stability behavior of atogepant and can be helpful for impurity profiling, API/formulation design, and quality control.