Efthymiou Tim, Krishnamurthy Ramanarayanan
Department of Chemistry, The Scripps Research Institute, La Jolla, California.
Curr Protoc Nucleic Acid Chem. 2015 Mar 9;60:2.19.1-2.19.20. doi: 10.1002/0471142700.nc0219s60.
Microwave-assisted chemical phosphitylation of novel nucleoside analogs containing a ribulose sugar unit was successful with yields ranging from 50% to 79% using 2-cyanoethyl-N,N-diisopropyl chlorophosphoramidite as the phosphitylating reagent. The resultant phosphoramidite products remained intact, with no signs of degradation over extended reaction times (up to 60 min) at an elevated temperature (65°C). When the same microwave-mediated phosphitylating protocols were applied to canonical DNA and RNA nucleoside monomers as substrates, using either 2-cyanoethyl-N,N,-diisopropyl chlorophosphoramidite or 2-cyanoethyl-N,N,N',N'-tetraisopropyl phosphane with an activator, 40% to 90% yields of DNA and RNA phosphoramidites were obtained within 10 to 15 min. These results demonstrate that microwave-assisted phosphitylation is an efficient alternative to standard phosphitylating conditions that can be expanded and refined to include a variety of substrates.
使用2-氰基乙基-N,N-二异丙基氯代亚磷酰胺作为磷酰化试剂，新型含核糖ulose糖单元的核苷类似物的微波辅助化学磷酰化反应成功实现，产率在50%至79%之间。所得的亚磷酰胺产物保持完整，在高温（65°C）下延长反应时间（长达60分钟）没有降解迹象。当将相同的微波介导磷酰化方案应用于标准DNA和RNA核苷单体作为底物时，使用2-氰基乙基-N,N-二异丙基氯代亚磷酰胺或2-氰基乙基-N,N,N',N'-四异丙基膦与活化剂，在10至15分钟内获得了40%至90%产率的DNA和RNA亚磷酰胺。这些结果表明，微波辅助磷酰化是标准磷酰化条件的一种有效替代方法，可以扩展和改进以包括各种底物。
