den Hartog J A, Wille G, Scheublin R A, van Boom J H
Biochemistry. 1982 Mar 2;21(5):1009-18. doi: 10.1021/bi00534a028.
The synthesis via a phosphotriester method of the octadecaribonucleotide AUG(UUC)5 (19) is reported. The octadecanucleotide is meant to serve as a synthetic messenger in a ribosomal protein synthesizing system. A fully protected octadecamer intermediate (18a) was prepared by a block coupling procedure. For the introduction of the desired 3'-5'-internucleotide bonds a 3'-O-(2,2,2-trichloroethyl 2-chlorophenyl phosphate) function was incorporated into the monomeric building blocks which were applied in the synthesis of 18a. Monomeric and oligomeric compounds with a thus protected 3'-O-phosphotriester function can be selectively deblocked to give 3'-O-phosphodiester derivatives suitable for condensation with 5'-hydroxyl (oligo)nucleotides. Conversion of fully protected oligomers to 5'-hydroxyl derivatives, suitable for further coupling at the 5' end, was effected by selective removal of the levulinyl function at the 5' end. The fully protected octadecanucleotide 18a was deblocked with fluoride ions, followed by ammonia and acid to give the required octadecamer 19. The condensing agent 1-(2,4,6-triisopropyl-phenylsulfonyl)-3-nitro-1,2,4-triazole, which was applied to effect the formation of fully protected 3'-5'-internucleotide phosphotriester functions, may give rise to side reactions with the heterocylic bases uracil and guanine. The consequences of these side reactions for the synthesis of octadecamer 19 are reported.
报道了通过磷酸三酯法合成十八聚核糖核苷酸AUG(UUC)5(19)。该十八聚核苷酸旨在作为核糖体蛋白质合成系统中的合成信使。通过逐步偶联程序制备了完全保护的十八聚体中间体(18a)。为了引入所需的3'-5'-核苷酸间键，将3'-O-(2,2,2-三氯乙基2-氯苯基磷酸酯)官能团引入用于合成18a的单体构建块中。具有如此保护的3'-O-磷酸三酯官能团的单体和寡聚化合物可以被选择性地脱保护，得到适合与5'-羟基(寡)核苷酸缩合的3'-O-磷酸二酯衍生物。通过选择性去除5'端的乙酰丙酮酰官能团，将完全保护的寡聚物转化为适合在5'端进一步偶联的5'-羟基衍生物。完全保护的十八聚核苷酸18a用氟离子脱保护，然后用氨和酸处理，得到所需的十八聚体19。用于形成完全保护的3'-5'-核苷酸间磷酸三酯官能团的缩合剂1-(2,4,6-三异丙基苯磺酰基)-3-硝基-1,2,4-三唑可能会与杂环碱基尿嘧啶和鸟嘌呤发生副反应。报道了这些副反应对十八聚体19合成的影响。
