Acridine- and cholesterol-derivatized solid supports for improved synthesis of 3'-modified oligonucleotides.
Acridine- and cholesterol-derivatized solid supports for improved synthesis of 3'-modified oligonucleotides.
复制标题
吖啶和胆固醇衍生的固体支持物，用于改进 3-修饰寡核苷酸的合成。
DOI：
10.1021/bc00010a005
复制
发表时间：
1991
影响因子：
4.7
通讯作者：
MeyerJr,RB
中科院分区：
文献类型：
--
作者：
Reed,MW;Adams,AD;Nelson,JS;MeyerJr,RB
描述了新的固体支持物，其允许直接合成在3'末端带有胆固醇或吖啶的寡核苷酸。通过还原 N-Cbz-羟基-L-脯氨酸制备立体化学限定的氨基二醇。该接头分子首先用所需的缀合物分子酰化，然后被保护为二甲氧基三苯甲基醚。剩余的仲羟基被琥珀酰化并固定在可控孔径玻璃载体上。使用标准亚磷酰胺偶联和脱保护条件从这些支持物制备 3'-修饰的寡脱氧核苷酸 (ODN)。由胆固醇氯甲酸酯和氨基二醇连接体制备胆固醇修饰的支持物。由具有不同长度连接臂的吖啶四氟苯酯制备了两种类型的吖啶修饰的固体载体。在 3'-衍生化 ODN 的替代合成中，这些活性酯也用于 3'-胺修饰 ODN 的酰化。进行热变性研究以确定不同连接臂对与互补 ODN 靶标杂交的影响。 3'-修饰的 ODN 的简便合成和纯化使得这些功能化固体支持物特别适用于制备带有这些和其他修饰的寡核苷酸。
New solid supports are described which allow the direct synthesis of oligonucleotides bearing either cholesterol or acridine at the 3'-terminus. A stereochemically defined amino diol was prepared by reduction of N-Cbz-hydroxy-L-proline. This linker molecule was first acylated with the desired conjugate molecule, then protected as the dimethoxytrityl ether. The remaining secondary hydroxyl group was succinylated and immobilized on a controlled-pore glass support. 3'-Modified oligodeoxynucleotides (ODNs) were prepared from these supports by using standard phosphoramidite coupling and deprotection conditions. A cholesterol-modified support was prepared from cholesterol chloroformate and the amino diol linker. Two types of acridine-modifiedsolid supports were prepared from acridine tetrafluorophenyl esters with linker arms of different length. In an alternative synthesis of 3'-derivatized ODNs, these active esters were also utilized for acylation of a 3'-amine-modified ODN. A thermal denaturation study was done to determine the effect of the different linker arms on hybridization to a complementary ODN target. Facile synthesis and purificationof the 3'-modified ODNs makes these functionalized solid supports especially useful for preparation of oligonucleotides bearing these and other modifications.