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    • 7. 发明授权
    • Modified nucleotides
    • 修饰的核苷酸
    • US08071739B2
    • 2011-12-06
    • US12804352
    • 2010-07-20
    • John MiltonXiaolin WuMark SmithJoseph BrennanColin BarnesXiaohai LiuSilke Ruediger
    • John MiltonXiaolin WuMark SmithJoseph BrennanColin BarnesXiaohai LiuSilke Ruediger
    • C07H21/02C07H21/04C07H19/04C12Q1/68
    • C12Q1/6869C07H1/00C07H19/06C07H19/10C07H19/14C07H19/16C07H19/20C07H21/00C12Q2525/186C12Q2535/113Y02P20/55C12Q2525/197
    • The invention provides modified nucleotide or nucleoside molecule comprising a purine or pyrimidine base and a ribose or deoxyribose sugar moiety having a removable 3′-OH blocking group covalently attached thereto, such that the 3′ carbon atom has attached a group of the structure —O—Z wherein Z is any of —C(R′)2-O—R″, —C(R′)2-N(R″)2, —C(R′)2-N(H)R″, —C(R′)2-S—R″ and —C(R′)2-F, wherein each R″ is or is part of a removable protecting group; each R′ is independently a hydrogen atom, an alkyl, substituted alkyl, arylalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclic, acyl, cyano, alkoxy, aryloxy, heteroaryloxy or amido group, or a detectable label attached through a linking group; or (R′)2 represents an alkylidene group of formula ═C(R′″)2 wherein each R′″ may be the same or different and is selected from the group comprising hydrogen and halogen atoms and alkyl groups; and wherein said molecule may be reacted to yield an intermediate in which each R″ is exchanged for H or, where Z is —C(R′)2-F, the F is exchanged for OH, SH or NH2, preferably OH, which intermediate dissociates under aqueous conditions to afford a molecule with a free 3′OH; with the proviso that where Z is —C(R′)2-S—R″, both R′ groups are not H.
    • 本发明提供了修饰的核苷酸或核苷分子,其包含嘌呤或嘧啶碱基和核糖或脱氧核糖糖部分,其具有共价连接到其上的可移除的3'-OH封闭基团,使得3'碳原子连接一组结构-O -Z其中Z是-C(R')2 -O-R“,-C(R')2 -N(R”)2,-C(R')2 -N(H)R“ -C(R')2 -S-R“和-C(R')2 -F,其中每个R”为或可移除的保护基团的一部分; 每个R'独立地是氢原子,烷基,取代的烷基,芳基烷基,烯基,炔基,芳基,杂芳基,杂环,酰基,氰基,烷氧基,芳氧基,杂芳氧基或酰氨基,或通过连接基团连接的可检测标记; 或(R')2表示式≡C(R'“)2的亚烷基,其中每个R”可以相同或不同,并且选自氢和卤素原子和烷基; 并且其中所述分子可以反应以产生其中每个R“被交换为H的中间体,或其中Z是-C(R')2 -F,F被交换成OH,SH或NH 2,优选OH,其中 中间体在水性条件下解离得到具有游离3'OH的分子; 条件是其中Z是-C(R')2 -S-R“,两个R'基团不是H.