Synthesis and Biochemical Evaluation of Phosphonoformate Oligodeoxyribonucleotides
作者:Christina M. Yamada、Douglas J. Dellinger、Marvin H. Caruthers
DOI:10.1021/ja060112b
日期:2006.4.1
acid linker. Synthons were sequentially added to this support using tetrazole as an activator, oxidized to phosphonoformate, and the transient 5'-protecting group was removed with acid. Following total synthesis of an oligomer, protecting groups were removed with TEMED.HF and products purified by HPLC. These analogues were resistant to nucleases, formed duplexes with complementary RNA (A-form), and,
膦甲酸寡脱氧核糖核苷酸是通过固相合成策略制备的。制备合适的合成子的第一步是在金属钠存在下将双(N,N-二异丙基氨基)膦和氯甲酸二苯基甲基甲硅烷基乙酯缩合,得到甲酸,[双(N,N-二异丙基氨基)膦基]-β-(二苯基甲基甲硅烷基乙基)酯。然后使用4,5-二氰基咪唑将该反应的产物与适当保护的2'-脱氧核苷缩合以产生3'-O-亚膦酰胺反应性单体。胞嘧啶、腺嘌呤和鸟嘌呤的环外胺用 9-芴基甲氧羰基保护,并使用氢醌-O,O'-二乙酸接头在受控孔玻璃上合成寡脱氧核糖核苷酸。使用四唑作为活化剂,将合成子依次添加到该载体中,氧化成膦甲酸酯,并用酸去除瞬时 5'-保护基团。在低聚物全合成后,用TEMED.HF去除保护基团并通过HPLC纯化产物。这些类似物对核酸酶具有抗性,与互补 RNA(A 型)形成双链体,并且作为在选定位点含有磷酸盐的嵌合寡聚体,刺激了 RNase H1 活性。