Tanaka T, Yamada Y, Sakata T, Uesugi S, Ikehara M
Faculty of Pharmaceutical Sciences, Osaka University, Japan.
Nucleic Acids Symp Ser. 1988(19):21-4.
A new phosphorylating agent, S-(N-monomethoxytritylaminoethyl)-O-(O-chlorophenyl) phosphorothioate, was prepared and reacted with a 5'-hydroxyl group of an oligonucleotide using 1-mesitylene-sulfonyl-3-nitrotriazole (MSNT) as a condensing agent. After base labile protecting groups were removed, the partially deprotected oligonucleotide was separated on a reversed phase column and converted to the oligonucleotide with an aminoethyl or a phosphoryl group at the 5'-end by treatment with 80% acetic acid or iodine-water, respectively. The syntheses of ppT, pppT, A5'pp5'T and A5'ppp5'T were also performed by treatment of 5'-O-(N-monomethoxytritylaminoethylthiophosphoryl) thymidine with tri-n-octylammonium salt of phosphoric acid, pyrophosphoric acid, pA and ppA, respectively.
制备了一种新的磷酸化试剂S-(N-单甲氧基三苯甲基氨基乙基)-O-(邻氯苯基)硫代磷酸酯，并以1-均三甲苯磺酰基-3-硝基三唑(MSNT)作为缩合剂，使其与寡核苷酸的5'-羟基反应。去除对碱不稳定的保护基团后，将部分脱保护的寡核苷酸在反相柱上分离，分别用80%乙酸或碘水处理，转化为5'-端带有氨乙基或磷酰基的寡核苷酸。通过分别用磷酸、焦磷酸、对氨基苯磷酸和焦磷酸对氨基苯磷酸的三正辛铵盐处理5'-O-(N-单甲氧基三苯甲基氨基乙基硫代磷酰基)胸苷，还进行了ppT、pppT、A5'pp5'T和A5'ppp5'T的合成。
