A novel thymidine phosphorylase to synthesize (halogenated) anticancer and antiviral nucleoside drugs in continuous flow

Four pharmaceutically relevant nucleoside analogues (5-fluoro-2 '-deoxyuridine, 5-chloro-2 '-deoxyuridine, 5-bromo-2 '-deoxyuridine, and 5-iodo-2 '-deoxyuridine) have been synthesized by using a novel thymidine phosphorylase from the halotolerant H. elongata (HeTP). Following enzyme immobilization on microbeads, the biocatalyst was implemented as a packed-bed reactor for the continuous production of halogenated nucleosides, achieving up to 90% conversion at the 10 mM scale with 30 min residence time. Taking the synthesis of floxuridine (5-fluoro-2 '-deoxyuridine) as a study case, we obtained the highest space-time yield (5.5 g L-1 h(-1)) reported to date. In addition, bioinformatic tools such as MD analysis and CapiPy have contributed to shine light on the catalytic performance of HeTP as well as its immobilization, respectively.

Automated summary

Four pharmaceutically relevant nucleoside analogues were synthesized using a novel thymidine phosphorylase from H. elongata. The enzyme was immobilized on microbeads and used as a packed-bed reactor for continuous production of halogenated nucleosides, achieving a high conversion rate. The highest space-time yield reported to date was obtained for the synthesis of floxuridine.