Abstract：
Mitapivat is a novel, first-in-class orally active pyruvate kinase activator approved by the US Food and Drug Administration in 2022 for the treatment of hemolytic anemia. There is no literature available regarding the identification of degradation impurities of mitapivat. The present study deals with the degradation behavior of mitapivat under various stress conditions such as hydrolytic, photolytic, thermal, and oxidative stress. The multivariate analysis found that the independent variables, that is, molarity, temperature, and time, are interacting with each other to affect the degradation of mitapivat. A specific, accurate, and precise high-performance liquid chromatographic (HPLC) method was developed to separate mitapivat from its degradation products. The separation was achieved on the C-18 column (250 mm × 4.6 mm × 5 µm) using the combination of 0.1% formic acid buffer and acetonitrile in gradient elution profile. The method was validated as per the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use Q2(R2) guideline. LC-electrospray ionization-Quadrupole-time of flight was employed to identify degradation products. A total of seven novel degradation products of mitapivat were identified based on tandem mass spectrometry and accurate mass measurement. In-silico toxicity of mitapivat and its degradation products was qualitatively evaluated by the DEREK toxicity prediction tool.
摘要：
Mitapivat是一部小说,2022年由美国食品和药物管理局批准用于治疗溶血性贫血的一流口服活性丙酮酸激酶激活剂。没有关于mitapivat降解杂质的鉴定的文献。本研究涉及mitapivat在各种胁迫条件下的降解行为，例如水解，光解，热,和氧化应激。多变量分析发现，自变量，也就是说,摩尔浓度,温度,和时间,彼此相互作用以影响mitapivat的降解。一个具体的，准确,并开发了精确的高效液相色谱（HPLC）方法来从其降解产物中分离mitapivat。在C-18柱（250mm×4.6mm×5µm）上使用0.1％甲酸缓冲液和乙腈的组合进行梯度洗脱。该方法按照国际人用药品技术要求协调理事会Q2(R2)指南进行验证。采用LC-电喷雾电离-四极杆-飞行时间来鉴定降解产物。基于串联质谱和准确的质量测量，总共鉴定了七种新型的米塔皮伐降解产物。通过DEREK毒性预测工具对mitapivat及其降解产物的硅内毒性进行了定性评估。
