Ikehara M, Oshie K, Hasegawa A, Ohtsuka E
Nucleic Acids Res. 1981 Apr 24;9(8):2003-20. doi: 10.1093/nar/9.8.2003.
2'-5'-Linked oligoadenylic acid 5'-triphosphates (2-5A) having chain lengths of 2-4 have been synthesized by polymerization of 3'-O-(o-nitrobenzyl)-N-benzoyladenosine 5'-phosphate followed by 5'-triphosphorylation via the imidazolidates. A large scale preparation of 5'-O-phosphoryladenylyl-(2'-5')-adenylyl-(2'-5')-adenosine was performed by the phosphotriester method using 5'-O-monomethoxytrityl-3'-O-(o-nitrobenzyl)-N-benzoyladenosine 2'-O-p-chlorophenylphosphate and 5'-O-phosphorodianilido-3'-O-(o-nitrobenzyl)-N-benzoyladenosine 2'-O-p-chlorophenylphosphate as intermediates. The trimer was also triphosphorylated by the imidazolide method. CD spectra for 5'-mono and triphosphorylated 2'-5' adenylates were measured as well as their UV hypochromicities. This triester method was also applied to the synthesis of 3',5'-bisphosphorylated protected oligoadenylic acids with natural 3'-5' linkages which could be used for further condensations to yield 5'-phosphorylated polynucleotides.
链长为2至4的2'-5'-连接的寡聚腺苷酸5'-三磷酸酯(2-5A)已通过3'-O-(邻硝基苄基)-N-苯甲酰腺苷5'-磷酸酯的聚合反应,然后通过咪唑酸酯进行5'-三磷酸化反应合成。通过使用5'-O-单甲氧基三苯甲基-3'-O-(邻硝基苄基)-N-苯甲酰腺苷2'-O-对氯苯基磷酸酯和5'-O-磷酰二苯胺基-3'-O-(邻硝基苄基)-N-苯甲酰腺苷2'-O-对氯苯基磷酸酯作为中间体的磷酸三酯法,大规模制备了5'-O-磷酰腺苷基-(2'-5')-腺苷基-(2'-5')-腺苷。该三聚体也通过咪唑酸酯法进行了三磷酸化。测量了5'-单磷酸化和三磷酸化的2'-5'腺苷酸的圆二色光谱及其紫外减色性。该磷酸三酯法还应用于合成具有天然3'-5'连接的3',5'-双磷酸化保护的寡聚腺苷酸,其可用于进一步缩合以产生5'-磷酸化的多核苷酸。