专利快速检索-快速检索全球专利，免费商用专利数据库-IPRDB

3. 发明申请

WO2013029801A1 MULTIPLE ORTHOGONAL LABELLING OF OLIGONUCLEOTIDES 审中-公开
标题翻译：寡核苷酸的多重正交标记
公开(公告)号：WO2013029801A1
公开(公告)日：2013-03-07
申请号：PCT/EP2012/003666
申请日：2012-08-31
申请人： DEUTSCHES KREBSFORSCHUNGSZENTRUM , RUPRECHT-KARLS-UNIVERSITÄT HEIDELBERG , SCHOCH, Juliane , JÄSCHKE, Andres , SAMANTA, Ayan , WIESSLER, Manfred
发明人： SCHOCH, Juliane , JÄSCHKE, Andres , SAMANTA, Ayan , WIESSLER, Manfred
IPC分类号： C07H21/00
CPC分类号： C07H21/00
摘要： In summary, the present invention concerns a method for multiple orthogonal labelling of oligonucleotides, preferably RNA or DNA, by simultaneously performing the inverse Diels-Alder reaction (DAinv) and the copper-catalyzed click reaction (CuAAC), wherein the method is employed in a single step by just adding the different reaction components together and incubating the aqueous reaction mixture preferably for one hour at room temperature. In detail, the reaction components are one or more N 3 -modified labels, a copper compound, a stabilizing ligand, a reducing agent and one or more electron-deficient label-modified dienes that are added together with an at least double-modified oligonucleotide having one more nucleotides containing one or more N3-reactive groups and one or more electron-rich dienophiles, wherein a terminal alkyne moiety is preferably used as N3-reactive group(s) and a frans-cyclooctene moiety or norbornene is preferably used as electron-rich dienophile(s), more preferably frans-cyclooctene. Therefore, the present invention provides a one-pot method for post-synthetic multiple orthogonal labeling of oligonucleotides, which allows the site-specific introduction of more than one label, preferably of at least two labels into oligonucleotides after solid-phase synthesis, wherein the DAinv takes place on the dienophile modification only and the CuAAC selectively takes place on the N3-reactive group modification.
摘要翻译：总之，本发明涉及通过同时进行反狄尔斯 - 阿尔德反应（DAinv）和铜催化的咔嗒反应（CuAAC）来对寡核苷酸（优选RNA或DNA）进行多重正交标记的方法，其中该方法用于通过仅将不同的反应组分加入一起并将该水性反应混合物在室温下温育1小时来进行单步骤。详细地，反应成分是与至少双修饰寡核苷酸一起加入的一种或多种N 3修饰标记，铜化合物，稳定配体，还原剂和一种或多种电子缺陷型标记修饰的二烯，另外还有一个含有一个或多个N 3 - 反应性基团和一个或多个富电子基性亲核体的核苷酸，其中末端炔部分优选用作N 3反应性基团，并且优选使用反式 - 环辛烯部分或降冰片烯作为电子 - 富含亲二烯体，更优选为反式环辛烯。因此，本发明提供了一种用于寡核苷酸的后合成多重正交标记的一锅法，其允许在固相合成之后将多于一个标记，优选至少两个标记的位点特异性引入寡核苷酸，其中 DAinv仅在亲双烯体修饰上发生，CuAAC选择性地发生在N 3反应性基团修饰上。

IPRDB

热门服务

关于我们

友情链接

联系方式