Solid-Phase Synthesis of Phosphorothioate Oligonucleotides Using Sulfurization Byproducts for in Situ Capping,The Journal of Organic Chemistry

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Solid-Phase Synthesis of Phosphorothioate Oligonucleotides Using Sulfurization Byproducts for in Situ Capping
The Journal of Organic Chemistry ( IF 3.3 ) Pub Date : 2018-09-04 00:00:00 , DOI: 10.1021/acs.joc.8b01553
Jimin Yang ₁ , Jessica A. Stolee ₁ , Hong Jiang ₁ , Li Xiao ₁ , William F. Kiesman ₁ , Firoz D. Antia ₁ , Yannick A. Fillon ₁ , Austen Ng ₁ , Xianglin Shi ₁

Affiliation

Oligonucleotides containing phosphorothioate (PS) linkages have recently demonstrated significant clinical utility. PS oligonucleotides are manufactured via a solid-phase chain elongation process in which a four-reaction cycle consisting of detritylation, coupling, sulfurization, and failure sequence capping with Ac₂O is repeated. In the capping step, uncoupled sequences are acetylated at the 5′–OH to stop the chain growth and control the levels of deletion, or (n–1), impurities. Herein, we report that the byproducts of commonly used sulfurization reagents react with the 5′–OH and cap the failure sequences. The standard Ac₂O capping step can therefore be eliminated, and this 3-reaction cycle process affords a higher yield and higher or comparable overall purity compared to the conventional 4-reaction synthesis. This improvement results in reducing the number of reactions from ∼80 to ∼60 for the synthesis of a typical length 20-mer oligonucleotide. For every kilogram of an oligonucleotide intermediate synthesized, > 500 L of reagents and organic solvents is saved, and the E-factor is decreased to <1500 from ∼2000.

中文翻译：

固相合成硫代磷酸寡核苷酸使用硫化副产物进行原位加帽

含有硫代磷酸酯（PS）键的寡核苷酸最近已证明具有重要的临床应用价值。PS寡核苷酸是通过固相链延伸过程制造的，在该过程中，重复了由去三苯甲基化，偶联，硫化和用Ac ₂ O封端的失败序列组成的四个反应循环。在封盖步骤中，未偶联的序列在5'–OH处被乙酰化，以终止链增长并控制缺失或（n –1）杂质的水平。在本文中，我们报道了常用的硫化试剂的副产物与5'-OH反应并限制了失效序列。标准Ac ₂因此，可以省去O封端步骤，并且与常规的4-反应合成相比，该3-反应循环过程提供了更高的产率和更高或相当的总纯度。该改进导致用于合成典型长度的20聚体寡核苷酸的反应数量从约80减少至约60。对于合成的每千克寡核苷酸中间体，可节省> 500 L的试剂和有机溶剂，并且E因子从约2000降低至<1500。

更新日期：2018-09-04

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