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    • 5. 发明申请
    • LINKER PHOSPHORAMIDITES FOR OLIGONUCLEOTIDE SYNTHESIS
    • 用于寡核苷酸合成的连接磷酸酰胺
    • WO0220537A3
    • 2002-09-06
    • PCT/CA0101263
    • 2001-09-10
    • UNIV TECHNOLOGIES INTPON RICHARD TYU SHUYUAN
    • PON RICHARD TYU SHUYUAN
    • C07H19/073C07B61/00C07H19/00C07H19/10C07H19/20C07H21/00C07H21/04C07H19/06C07H19/16
    • C07H19/10C07B2200/11C07H19/00C07H19/20C07H21/00C40B40/00
    • A novel approach for combining the ease of cleavage of carboxylic acid linker arms with the single phosphoramidite coupling chemistry of the universal supports useful in oligonucleotide synthesis. There is disclosed a new class of phosphoramidite reagents, linker phosphoramidites, which contain a bifunctional linker arm with a protected nucleoside linked through a 3'-ester bond on one end and a reactive phosphoramidite group or other phosphate precursor group on the other end. The phosphoramidite group on the linker phosphoramidite may be activated under the same conditions and has similar reactivity as conventional nucleoside-3'-phosphoramidite reagents lacking the intermediate linker arm. The 3'-ester linkage contained within the linker phosphoramidite has similar properties to the linkages on prederivatized supports. The ester linkage is stable to all subsequent synthesis steps, but upon treatment with a cleavage reagent, such as ammonium hydroxide, the ester linkage is hydrolyzed. This releases the oligonucleotide product with the desired 3'-hydroxyl terminus and leaves the phosphate portion of the reagent attached to the support, which is subsequently discarded.
    • 将羧酸接头臂的易切割性与用于寡核苷酸合成的通用载体的单一亚磷酰胺偶联化学结合起来的新方法。 公开了一类亚磷酰胺试剂,连接亚磷酰胺,其含有双功能连接臂,其具有通过一端上的3'-酯键连接的被保护的核苷酸和另一端的反应性亚磷酰胺基团或其它磷酸酯前体基团。 连接亚磷酰胺上的亚磷酰胺基团可以在相同条件下被活化,并且与缺少中间连接臂的常规核苷-3'-亚磷酰胺试剂具有相似的反应性。 连接子亚磷酰胺中含有的3'-酯键具有与预衍生载体上的连接相似的性质。 酯键对所有随后的合成步骤是稳定的,但是当用切割试剂如氢氧化铵处理时,酯键被水解。 这释放出具有所需的3'-羟基末端的寡核苷酸产物并留下附着在载体上的试剂的磷酸部分,随后将其弃去。