申请人:THE PERKIN-ELMER CORPORATION
公开号:EP0786468A2
公开(公告)日:1997-07-30
The compounds of the invention are exemplified by the class of diglycolate synthesis supports particularly useful as support reagents for the direct synthesis of 3'-labeled polynucleotides. Generally, the compounds of the invention have the structure
where T is an acid-cleavable hydroxyl protecting group, e.g., 4,4'-dimethoxytrityl; L1 is a linker for connecting a 3'-terminal nitrogen to carbon; L2 and L3 are linkers for connecting oxygen and carbon; W is a solid support, e.g., CPG or polystyrene; L4 is a linker for connecting the solid support to nitrogen; R1 and R2 are nitrogen substituents, e.g., hydrogen, lower alkyl, nitrogen protecting group, or label; and R3 through R7 are carbon substituents, e.g., hydrogen or lower alkyl. In a first particularly preferred embodiment, the synthesis supports of the invention are exemplified by compounds having the structure
where DMT is 4,4'-dimethoxytrityl and W is polystyrene. In a second particularly preferred embodiment, the synthesis supports of the invention are exemplified by compounds having the structure
where DMT and W are defined above.
本发明的化合物属于二羟基乙酸酯合成支持物,特别适用于直接合成 3'- 标记的多核苷酸。一般来说,本发明的化合物具有如下结构
其中,T 是可被酸水解的羟基保护基团,如 4,4'-二甲氧基三苯甲基;L1 是连接 3'末端氮和碳的连接体;L2 和 L3 是连接氧和碳的连接体;W 是固体支持物,如 CPG 或聚苯乙烯;L2 和 L3 是连接氧和碳的连接体、R1和R2是氮取代基,如氢、低级烷基、氮保护基团或标签;R3至R7是碳取代基,如氢或低级烷基。在第一个特别优选的实施方案中,本发明的合成支持物以具有以下结构的化合物为例
其中 DMT 为 4,4'-二甲氧基三苯甲基,W 为聚苯乙烯。在第二个特别优选的实施方案中,本发明的合成支持物以具有以下结构的化合物为例
其中 DMT 和 W 定义如上。