James M. Wood, Dan G. Torres, Roselis A. Landaeta Aponte, Sandesh Deshpande, Alice M. McSweeney, Rinu Shrestha, Andrew Reddy, Gary B. Evans, Steven C. Almo, Tyler L. Grove, Richard L. Kingston, Vernon K. Ward, Lawrence D. Harris
3'-Deoxy-3',4'-didehydronucleoside triphosphates (ddhNTPs) are antiviral metabolites produced by viperin and viperin-like enzymes, all of which are members of the radical SAM superfamily of enzymes. To date, synthetic access to ddhUTP, ddhGTP, and ddhATP has remained limited. We report a scalable synthesis of the complete ddhN nucleoside family using Bobbitt's salt-mediated oxidation of nucleoside orthoformates, including a chromatography-free route to ddhC on a gram scale. This methodology enabled the first synthesis and evaluation of ddhUTP, ddhGTP, and ddhATP as inhibitors of RNA-dependent RNA polymerases, with ddhATP confirmed to act as an obligate chain terminator through primer extension assays.