The generation of bioactive molecules from inactive precursors is ac rucial step in the chemical evolution of life, however,mechanistic insights into this aspect of abiogenesis are scarce.Here,weinvestigate the protein-catalyzed formation of antivirals by the 3C-protease of enterovirusD 68. The enzyme induces aldol condensations yielding inhibitors with antiviral activity in cells.K inetic and thermodynamic analyses reveal that the bioactivity emerges from ad ynamic reaction system including inhibitor formation, alkylation of the protein target by the inhibitors,a nd competitive addition of non-protein nucleophiles to the inhibitors.T he most active antivirals are slowlyr eversible inhibitors with elongated target residence times.T he study reveals first examples for the chemical evolution of bio-actives through protein-catalyzed, non-enzymatic CÀCcouplings.The discoveredmechanism works under physiological conditions and might constitute anative process of drug development.

SARS coronavirus main proteases (3CL proteases) have been validated as pharmacological targets for the treatment of coronavirus infections. Current inhibitors of SARS main protease, including the clinically admitted drug nirmatrelvir are peptidomimetics with the downsides of this class of drugs including limited oral bioavailability, cellular permeability, and rapid metabolic degradation. Here, we investigate covalent fragment inhibitors of SARS Mpro as potential alternatives to peptidomimetic inhibitors in use today. Starting from inhibitors acylating the enzyme's active site, a set of reactive fragments was synthesized, and the inhibitory potency was correlated with the chemical stability of the inhibitors and the kinetic stability of the covalent enzyme‐inhibitor complex. We found that all tested acylating carboxylates, several of them published prominently, were hydrolyzed in assay buffer and the inhibitory acyl‐enzyme complexes were rapidly degraded leading to the irreversible inactivation of these drugs. Acylating carbonates were found to be more stable than acylating carboxylates, however, were inactive in infected cells. Finally, reversibly covalent fragments were investigated as chemically stable SARS CoV‐2 inhibitors. Best was a pyridine‐aldehyde fragment with an IC50 of 1.8 μM at a molecular weight of 211 g/mol, showing that pyridine fragments indeed are able to block the active site of SARS‐CoV‐2 main protease.

Leuven (Belgien) [ + + ]D iese Autoren haben zu gleichen Teilen zu der Arbeit beigetragen. [ + ++ + ]g eteilte Erstautorschaft Hintergrundinformationen und die Identifikationsnummern (OR-CID) der Autoren sind unter:https://doi.org/10.1002/ange. 202102074 zu finden. 2021 Die Autoren. AngewandteChemie verçffentlichtv on Wiley-VCH GmbH. Dieser Open Access Beitrag steht unter den Bedingungen der Creative Commons AttributionN on-CommercialN o-Derivs License, die eine Nutzung und Verbreitung in allen Medien gestattet, sofern der ursprüngliche Beitrag ordnungsgemäßzitiert und nicht fürkommerzielle Zwecke genutzt wird und keine ¾nderungen und Anpassungen vorgenommenwerden.