Optimization of solid-phase synthesis process of phosphorodiamidate morpholino oligonucleotides
10.7644/j.issn.1674-9960.2024.09.004
- VernacularTitle:磷酰二胺吗啉代寡核苷酸固相合成工艺优化
- Author:
Renjie LONG
1
;
Baogang WANG
;
Yabin SONG
Author Information
1. 安徽医科大学,合肥 230032
- Keywords:
solid-phase synthesis;
phosphorodiamidate morpholino oligonucleotides;
process optimization
- From:
Military Medical Sciences
2024;48(9):664-670
- CountryChina
- Language:Chinese
-
Abstract:
Objective To optimize the solid-phase synthesis process of phosphorodiamidate morpholino oligonucleotide(PMO)and determine the optimal reaction conditions.Methods The PMO tetramer PMO-TTTT was synthesized according to the reported reaction conditions,followed by purification through a semi-preparative high-performance liquid chromatography(HPLC)process.PMO-TTTT was structurally verified using nuclear magnetic resonance(NMR)and high-resolution mass spectrometry.With purified PMO-TTTT as a reference,a calibration curve was established,which subsequently guided the optimization of the reaction conditions for the solid-phase coupling reaction process,including the organic base,additives,duration of reaction and temperature.Under the optimized reaction condition,the anti-influenza A virus PMO sequence,PMO-flu,was synthesized and purified using a nucleic acid purification device.Results The optimal parameters for PMO solid-phase synthesis were determined.The organic base was N-ethylmorpholine,the additive was lithium iodide,the best temperature was 30 ℃,and the duration was 90 minutes.Conclusion The PMO solid-phase synthesis process has been established.LiI has been screened as a potent coupling reaction additive which could significantly boosts the efficiency of PMO solid-phase synthesis.
