Engineering a Cytidine Aminotransferase for Biocatalytic Production of the Antiviral Molnupiravir

• DOI: 10.26434/chemrxiv-2021-vfkqb-v2
• Publication Date: 2022-02-01T15:04:21Z
• Authors: Ashleigh Burke; William Birmingham; Ying Zhuo; Bruna Zuculoto da...; Rebecca Crawshaw; Thomas Thorpe; Ian Rowles; James Finnigan; Simon J. Charnock; Sarah Lovelock; Nicholas Turner; Anthony Green
• Keywords: Cytidine; Biocatalysis; Nucleotide salvage

The COVID-19 pandemic highlights the urgent need for cost-effective processes to rapidly manufacture antiviral drugs at scale. Here we report a concise biocatalytic process Molnupiravir, nucleoside analogue currently in phase 3 clinical trials as an orally available treatment SARS-CoV-2. Key success of this was development cytidine aminotransferase production N-hydroxy-cytidine through evolutionary adaption hydrolytic enzyme deaminase. This engineered biocatalyst performs >100,000 turnovers less than 30 minutes, operates 180 g/L substrate loading and benefits from situ crystallization product (>90% yield), which can be converted Molnupiravir by selective 5’-acylation using Novozym® 435.