El Zahar N M, Magdy N, El-Kosasy A M, Bartlett Michael G
Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt.
Department of Pharmaceutical and Biomedical Sciences, College of Pharmacy, University of Georgia, Athens, Georgia, USA.
Biomed Chromatogr. 2018 Jan;32(1). doi: 10.1002/bmc.4088. Epub 2017 Oct 4.
Phosphorothioate (PS) oligonucleotides are a rapidly rising class of drugs with significant therapeutic applications. However, owing to their complex structure and multistep synthesis and purification processes, generation of low-level impurities and degradation products are common. Therefore, they require significant investment in quality control and impurity identification. This requires the development of advanced methods for analysis, characterization and quantitation. In addition, the presence of the PS linkage leads to the formation of chiral centers which can affect their biological properties and therapeutic efficiency. In this review, the different types of oligonucleotide impurities and degradation products, with an emphasis on their origin, mechanism of formation and methods to reduce, prevent or even eliminate their production, will be extensively discussed. This review will focus mainly on the application of chromatographic techniques to determine these impurities but will also discuss other approaches such as mass spectrometry, capillary electrophoresis and nuclear magnetic resonance spectroscopy. Finally, the chirality and formation of diastereomer mixtures of PS oligonucleotides will be covered as well as approaches used for their characterization and the application for the development of stereochemically-controlled PS oligonucleotides.
硫代磷酸酯（PS）寡核苷酸是一类迅速崛起的药物，具有重要的治疗应用。然而，由于其结构复杂以及多步合成和纯化过程，低水平杂质和降解产物的产生很常见。因此，它们需要在质量控制和杂质鉴定方面投入大量资金。这就需要开发先进的分析、表征和定量方法。此外，PS键的存在会导致手性中心的形成，这可能会影响它们的生物学性质和治疗效果。在本综述中，将广泛讨论不同类型的寡核苷酸杂质和降解产物，重点是它们的来源、形成机制以及减少、预防甚至消除其产生的方法。本综述将主要关注色谱技术在测定这些杂质方面的应用，但也会讨论其他方法，如质谱、毛细管电泳和核磁共振光谱。最后，将涵盖PS寡核苷酸的手性和非对映异构体混合物的形成，以及用于其表征的方法和立体化学控制的PS寡核苷酸开发中的应用。
