Quality by Design-Assisted Separation and Characterization of Photolytic Degradation Products of Midostaurin Using
Rupali Mahajan1,2, Saurabh Shah3, Saurabh Srivastava3
1Department of Pharmaceutical Analysis, National Institute of Pharmaceutical Education and Research (NIPER), Hyderabad, India.
Rationale:
Midostaurin is the first-in-class antineoplastic agent for acute myeloid leukemia (AML), exhibits a distinctive pyrrolo[2,3-b]pyridine scaffold, and is susceptible to forming five forced degradation products under photolytic conditions. Impurities in products can compromise their safety and efficacy, often leading to significant recalls. Hence, strict quality control and reliable analytical methods are essential to prevent these risks. To the best of our knowledge, the lack of available literature on midostaurin in photolytic stress conditions drives the rationale for this study.
Methods:
The current study described a quality-by-design approach for developing a robust RP-HPLC analytical methodology. Simultaneously, we devised a rapid UHPLC technique that is transferable for LC-MS analysis to characterize DPs within a 6-min period using LC-Q-TOF-MS/MS. In QbD, a Taguchi orthogonal array design was employed for screening the significant factors, i.e., column temperature, buffer pH, and flow rate. A Box-Behnken design was employed to study the interactions of the chosen factors on the tailing factor and resolution of the analyte and five DPs. Additionally, Zeneth software was used to estimate photolytic DPs. Further, the study was extended using Derek and Sarah software to determine toxicity.
Results:
Midostaurin was found to form five degradation products under photolytic stress conditions. Four degradation products (DP-2, DP-3, DP-4, and DP-5; m/z 587, m/z 546, and m/z 585) were structurally characterized using LC-Q-TOF-MS/MS, whereas DP-1 was detected as an unidentified degradation peak due to a lack of ionization. Partial agreement was observed between Zeneth predictions and experimental findings. In silico toxicity assessment using Derek and Sarah software provided preliminary safety insights for the characterized degradation products.
Conclusions:
The study demonstrates the development of a QbD-based RP-HPLC and UHPLC-LC-MS method for the analysis of photolytic degradation products of midostaurin. The approach enables effective impurity profiling and provides supportive insights into degradation behavior and toxicity, contributing to improved quality control.
