Microwave-Assisted One-Step Synthesis of 2',3'-Cyclic Phosphates of Nucleosides

2',3'-Cyclic canonical nucleotides are an important class of compounds playing broad roles in regulating biological processes and are investigated in the context of prebiotic chemistry as activated nucleotides for oligonucleotide formation. Despite their growing importance, synthetic access of 2',3'-cyclic nucleotides is constrained, resulting in their cost-prohibitive commercial prices. Here, we describe a microwave-assisted one-pot synthesis starting from commercially available nucleosides employing an easily available cyclophosphorylating reagent, bis(dimethyldiamino)phosphorodiamidate (BDMDAP). The corresponding 2',3'-cyclic nucleotides are isolated in good yields (70-91%) by a simple ion-exchange column with no further workup. The nucleosides require no protecting group as the cyclophosphorylation reaction is selective for the 2',3'-dihydroxyl groups. The experimental protocol is robust and can be run in parallel to provide access to gram quantities of these 2',3'-cyclic nucleotides within a day. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Procedure for the phosphorylation of nucleoside (2',3'-cyclic phosphates of nucleosides) Support Protocol: Synthesis of bis(dimethylamino)phosphorodiamidate sodium salt.