Rapid Esterification of Nucleosides to Solid-Phase Supports for Oligonucleotide Synthesis Using Uronium and Phosphonium Coupling Reagents

• Published in: Bioconjugate chemistry 1999-11, Vol.10 (6), p.1051-1057
• Main Authors: Pon, Richard T, Yu, Shuyuan, Sanghvi, Yogesh S
• Format: Article
• Language: English
• Subjects: 4-Aminopyridine - analogs & derivatives; 4-Aminopyridine - chemistry; DNA - chemical synthesis; Esterification; Esters - chemistry; Imidazoles - chemistry; Indicators and Reagents; Nucleosides - chemistry; Oligonucleotides - chemical synthesis; Phosphorus Compounds - chemistry; Triazoles - chemistry; Urea - chemistry
• ISSN: 1043-1802; 1520-4812
• DOI: 10.1021/bc990063a

Nucleosides can be esterified to solid-phase supports using uronium or phosphonium coupling reagents and a coupling additive, such as 1-hydroxybenzotriazole (HOBT), 7-aza-1-hydroxybenzotriazole (HOAT), N-methylimidazole (NMI), or 4-(dimethylamino)pyridine (DMAP). However, DMAP was far superior to other additives and high nucleoside loadings (up to 60 μmol/g) and rapid coupling reactions (≤10 min) were possible. Hydroxyl-derivatized CPG was attached to nucleosides with 3‘-succinyl or 3‘-hydroquinone-O,O‘-diacetic acid (HQDA or Q-Linker) carboxyl groups through a primary ester linkage. Alternatively, supports derivatized with succinic acid or the Q-Linker were attached directly to the 3‘-OH group of nucleosides through a secondary ester linkage. Uronium reagents (HATU or HBTU) gave the best results with the HQDA linker arm, while the bromophosphonium (BrOP or PyBrOP) reagents were best with the succinyl linker arm. In all cases, the coupling reactions were much faster than previous methods using carbodiimide coupling reagents. The ease and speed of the reaction make this support derivatization procedure suitable for automated in situ couplings on DNA synthesizers.