Estalayo-Adriàn Sandra, Lartia Rémy, Meyer Albert, Vasseur Jean-Jacques, Morvan François, Defrancq Eric
Département de Chimie Moléculaire UMR CNRS 5250, Université Grenoble Alpes 38041, Grenoble Cedex 9, France.
Institut des Biomolécules Max Mousseron, UMR 5247 CNRS, Université de Montpellier 34095, Montpellier Cedex 5, France.
ChemistryOpen. 2015 Apr;4(2):169-73. doi: 10.1002/open.201402099. Epub 2014 Dec 10.
The conjugation of oligonucleotides with reporters is of great interest for improving their intrinsic properties or endowing new ones. In this context, we report herein a new procedure for the bis-labelling of oligonucleotides through oxime ligation (Click-O) and copper(I)-catalyzed alkyne-azide cycloaddition (Click-H). 5'-Azido and 3'-aldehyde precursors were incorporated into oligonucleotides, and subsequent coupling reactions through Click-O and Click-H (or vice versa) were successfully achieved. In particular, we exhaustively investigated the full compatibility of each required step for both tethering strategies. The results demonstrate that click Huisgen and click oxime reactions are fully compatible. However, whilst both approaches can deliver the targeted doubly conjugated oligonucleotide, the route involving click oxime ligation prior to click Huisgen is significantly more successful. Thus the reactions investigated here can be considered to be key elements of the chemical toolbox for the synthesis of highly sophisticated bioconjugates.
寡核苷酸与报告基团的共轭对于改善其固有性质或赋予新性质具有极大的意义。在此背景下，我们在此报告一种通过肟连接（Click - O）和铜（I）催化的炔 - 叠氮环加成反应（Click - H）对寡核苷酸进行双标记的新方法。将5'-叠氮基和3'-醛前体引入寡核苷酸中，并通过Click - O和Click - H（或反之亦然）成功实现了后续的偶联反应。特别是，我们详尽地研究了两种连接策略中每个所需步骤的完全兼容性。结果表明，点击式惠斯根反应和点击式肟反应完全兼容。然而，虽然两种方法都可以得到目标双共轭寡核苷酸，但在点击式惠斯根反应之前涉及点击式肟连接的路线明显更成功。因此，这里研究的反应可被视为合成高度复杂生物共轭物的化学工具箱中的关键要素。
