Oligonucleotide synthesis under mild deprotection conditions
Oligonucleotide synthesis under mild deprotection conditions
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温和脱保护条件下的寡核苷酸合成
DOI：
10.1039/d2nj03845e
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发表时间：
2023
影响因子：
3.3
通讯作者：
Fang, Shiyue
中科院分区：
文献类型：
--
作者：
Chillar, Komal;Eriyagama, Adikari M.;Yin, Yipeng;Shahsavari, Shahien;Halami, Bhaskar;Apostle, Alexander;Fang, Shiyue
在DNA和RNA中发现了超过100种非规范核苷酸。它们中的许多对亲核试剂敏感。由于已知的寡核苷酸合成技术需要亲核条件进行脱保护，因此目前没有合适的技术用于其合成。最近公开的关于使用1，3-二噻烷-2-基-甲基（Dim）进行磷酸酯保护和使用1，3-二噻烷-2-基-甲氧羰基（Dmoc）进行氨基保护的方法可以解决该问题。在Dim-Dmoc保护下，可以用NaIO 4然后用苯胺实现寡脱氧核苷酸（ODN）脱保护。一些敏感基团已被确定在这些条件下是稳定的。除了用作碱之外，苯胺还用作亲核清除剂，其防止脱保护副产物与ODN反应。因此，需要过量的苯胺。在这里，我们报告了使用烷基Dim（aDim）和烷基Dmoc（aDmoc）用于ODN合成。在aDim-aDmoc保护的情况下，用NaIO 4随后用K2 CO 3实现脱保护。不需要亲核清除剂如苯胺。合成了10多个ODNs，其中包括一个含有高度敏感的N4-乙酰胞苷的ODNs。将该方法扩展到敏感RNA合成的工作正在进行中。
Over a hundred non-canonical nucleotides have been found in DNA and RNA. Many of them are sensitive toward nucleophiles. Because known oligonucleotide synthesis technologies require nucleophilic conditions for deprotection, currently there is no suitable technology for their synthesis. The recently disclosed method regarding the use of 1,3-dithian-2-yl-methyl (Dim) for phosphate protection and 1,3-dithian-2-yl-methoxycarbonyl (Dmoc) for amino protection can solve the problem. With Dim–Dmoc protection, oligodeoxynucleotide (ODN) deprotection can be achieved with NaIO4 followed by aniline. Some sensitive groups have been determined to be stable under these conditions. Besides serving as a base, aniline also serves as a nucleophilic scavenger, which prevents deprotection side products from reacting with ODN. For this reason, excess aniline is needed. Here, we report the use of alkyl Dim (aDim) and alkyl Dmoc (aDmoc) for ODN synthesis. With aDim–aDmoc protection, deprotection is achieved with NaIO4 followed by K2CO3. No nucleophilic scavenger such as aniline is needed. Over 10 ODNs including one containing the highly sensitive N4-acetylcytidine were synthesized. Work on extending the method for sensitive RNA synthesis is in progress.