Department of Pharmaceutical Analysis, National Institute of Pharmaceutical Education and Research-Ahmedabad (Ministry of Chemicals and Fertilizers, Government of India), Opposite Air Force Station, Palaj, India.
Biomed Chromatogr. 2024 Oct;38(10):e5983. doi: 10.1002/bmc.5983. Epub 2024 Aug 7.
Peptide aggregation is one of the key challenges associated with the development of therapeutic peptides. Peptide and protein aggregates are considered as one of the most important critical quality attributes (CQA). Therapeutic liraglutide (LGT) is proteinaceous in nature, and aggregation can be triggered by various environmental stress condition. Therefore, it is essential to separate and identify aggregation states of such drugs. In this study, we have established size exclusion chromatography-liquid chromatography-ultraviolet/high resolution mass spectrometry (SEC-LC-UV/HRMS) method to separate and identify the stress induced LGT aggregates. LGT samples were subjected to photolytic, thermal, freeze thaw and shaking stress conditions. Additionally, LGT solution was incubated with surfactant and excipient that are commonly used in peptide formulation, to evaluate their impact on aggregation level and physicochemical stability over time. The developed SEC method was also validated for specificity, accuracy, precision and linearity. The results of this study will be useful for investigators to monitor LGT aggregates during product development.
肽聚集是与治疗性肽的开发相关的关键挑战之一。肽和蛋白质聚集物被认为是最重要的关键质量属性（CQA）之一。治疗性利拉鲁肽（LGT）本质上是蛋白质，并且聚集可以由各种环境应激条件引发。因此，分离和鉴定此类药物的聚集状态至关重要。在这项研究中，我们建立了尺寸排阻色谱-液相色谱-紫外/高分辨质谱（SEC-LC-UV/HRMS）方法来分离和鉴定应激诱导的 LGT 聚集物。将 LGT 样品置于光解、热、冻融和振荡应激条件下。此外，还将 LGT 溶液与肽制剂中常用的表面活性剂和赋形剂孵育，以评估它们对聚集水平和物理化学稳定性随时间的影响。还对开发的 SEC 方法进行了专属性、准确性、精密度和线性验证。这项研究的结果将有助于研究人员在产品开发过程中监测 LGT 聚集物。
