Chen Tao, Tang Shijia, Fu Yige, Napolitano José G, Zhang Kelly
Small Molecule Analytical Chemistry, Small Molecule Pharmaceutical Sciences, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, United States.
Small Molecule Analytical Chemistry, Small Molecule Pharmaceutical Sciences, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, United States.
J Chromatogr A. 2022 Aug 16;1678:463349. doi: 10.1016/j.chroma.2022.463349. Epub 2022 Jul 16.
Oligonucleotides have emerged as powerful therapeutics for treating diverse diseases. To fully unlock the therapeutic potential of oligonucleotides, there is still a great need to further improve their drug-like properties. Numerous chemical modifications have been explored to achieve this goal, with phosphorothioation being one of the most widely used strategies. However, phosphorothioate modification produces diastereomers that are reported to have different properties and performances, demanding detailed characterization of these diastereomers. Here we provide an overview of phosphorothioated oligonucleotide diastereomers, covering their origin and configurations, physicochemical and pharmacological properties, and stereo-selective chemical synthesis, followed by a summary of currently available analytical techniques for characterizing these diastereomers, with a focus on liquid chromatography-based approaches, including ion-pair reversed-phase liquid chromatography, anion exchange chromatography, mixed-mode chromatography, and hybrid approaches. Non-chromatographic techniques, such as capillary electrophoresis, spectroscopy and other methods, are also being reviewed.
寡核苷酸已成为治疗多种疾病的有效疗法。为了充分释放寡核苷酸的治疗潜力，仍迫切需要进一步改善其类药物性质。人们已探索了许多化学修饰方法来实现这一目标，硫代磷酸化是应用最广泛的策略之一。然而，硫代磷酸酯修饰会产生非对映异构体，据报道这些异构体具有不同的性质和性能，因此需要对这些非对映异构体进行详细表征。在此，我们概述了硫代磷酸化寡核苷酸非对映异构体，涵盖其来源和构型、物理化学和药理学性质以及立体选择性化学合成，随后总结了目前用于表征这些非对映异构体的分析技术，重点介绍基于液相色谱的方法，包括离子对反相液相色谱、阴离子交换色谱、混合模式色谱和混合方法。还对非色谱技术，如毛细管电泳、光谱学和其他方法进行了综述。
