Kore Anilkumar R, Yang Bo, Srinivasan Balasubramanian
Life Sciences Solutions Group, Thermo Fisher Scientific, Austin, Texas.
Curr Protoc Nucleic Acid Chem. 2015 Mar 9;60:1.33.1-1.33.10. doi: 10.1002/0471142700.nc0133s60.
An efficient, reliable method for the chemical synthesis of (E)-5-[3-aminoallyl]-uridine-5'-O-triphosphate (AA-UTP), starting from 5-iodouridine, is described. This new strategy features the involvement of one-pot triphosphate formation and fluorous solid-phase extraction (F-SPE). The one-pot synthesis involves the mono phosphorylation of fluorous-tagged uridine, followed by the reaction with pyrophosphate to afford the fluorous-tagged AA-UTP. The F-SPE is achieved by installing a fluorous-tag onto the uridine prior to triphosphate formation, purification via F-SPE, and cleavage of the fluorous-tag. It is worth mentioning that this protocol produces AA-UTP in high yield and purity using one simple F-SPE; no conventional column chromatography is involved.
本文描述了一种从5-碘尿苷开始，高效、可靠地化学合成（E）-5-[3-氨基烯丙基]-尿苷-5'-O-三磷酸（AA-UTP）的方法。这种新策略的特点是一锅法形成三磷酸酯和氟固相萃取（F-SPE）。一锅法合成包括对氟标记的尿苷进行单磷酸化，然后与焦磷酸反应生成氟标记的AA-UTP。F-SPE是通过在形成三磷酸酯之前在尿苷上安装一个氟标记，通过F-SPE进行纯化，以及去除氟标记来实现的。值得一提的是，该方案使用一种简单的F-SPE就能以高产率和高纯度生产AA-UTP；无需传统的柱色谱法。
