申请人:Hokkaido System Science Co., Ltd.
公开号:EP2857412A1
公开(公告)日:2015-04-08
A nucleic acid synthesis method enabling a reaction in a fluid (flow) with a highly dispersible liquid-phase support to improve coupling efficiency is provided.
The method for synthesizing an oligonucleotide comprising: sequentially condensing and oxidizing a nucleoside phosphoramidite compound in the presence of an acid/azole complex compound using a starting raw material, i.e., hydrophobic group-bonded nucleoside represented by Formula (1):
where R1: an alkylene group having 1 to 12 carbon atoms,
R2: an alkylene group having 1 to 22 carbon atoms, R3 and
R4 each independently represent an alkyl group having 1 to 22 carbon atoms or the like, R5: a single bond or an alkylene group having 1 to 22 carbon atoms, R6: each independently an alkyl group having 6 to 30 carbon atoms, n represents an integer of 2 to 6, X represents a hydrogen atom, hydroxyl group, or the like, Y: a protecting group deprotectable under an acidic condition, and Z: an adenyl group, a guanyl group, or the like having a polar group optionally protected by a protecting group,
wherein a condensation reaction is performed by preliminarily dissolving the hydrophobic group-bonded nucleoside or hydrophobic group-bonded oligonucleotide and the nucleoside phosphoramidite compound in a nonpolar solvent, and contacting the resulting solution with the acid/azole complex compound or a solution containing the complex compound.
本发明提供了一种核酸合成方法,可在具有高分散性液相支持物的流体(流动)中进行反应,从而提高耦合效率。
该方法用于合成寡核苷酸,包括:使用起始原料,即由式(1)表示的疏水基团键合核苷,在酸/唑复合物存在下,依次缩合和氧化核苷磷酰胺化合物:
其中 R1:具有 1 至 12 个碳原子的亚烷基、
R2:具有 1 至 22 个碳原子的亚烷基, R3 和
R4:各自独立地代表具有 1 至 22 个碳原子的烷基或类似基团; R5:单键或具有 1 至 22 个碳原子的亚烷基; R6:各自独立地代表具有 6 至 30 个碳原子的烷基; n 代表 2 至 6 的整数; X 代表氢原子、羟基或类似基团; Y:可在酸性条件下脱保护的保护基团; Z:具有极性基团的腺嘌呤基、鸟苷酸基或类似基团,可选择受保护基团的保护、
其中缩合反应是通过在非极性溶剂中初步溶解疏水基团键合的核苷或疏水基团键合的寡核苷酸和核苷亚磷酰胺化合物,并将所得溶液与酸/唑复合物或含有复合物的溶液接触来进行的。