RT Journal Article
SR Electronic
T1 Alpha-ketoamides as broad-spectrum inhibitors of coronavirus and enterovirus replication
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.02.10.936898
DO 10.1101/2020.02.10.936898
A1 Linlin Zhang
A1 Daizong Lin
A1 Yuri Kusov
A1 Yong Nian
A1 Qingjun Ma
A1 Jiang Wang
A1 Albrecht von Brunn
A1 Pieter Leyssen
A1 Kristina Lanko
A1 Johan Neyts
A1 Adriaan de Wilde
A1 Eric J. Snijder
A1 Hong Liu
A1 Rolf Hilgenfeld
YR 2020
UL http://biorxiv.org/content/early/2020/02/10/2020.02.10.936898.abstract
AB The main protease of coronaviruses and the 3C protease of enteroviruses share a similar active-site architecture and a unique requirement for glutamine in the P1 position of the substrate. Because of their unique specificity and essential role in viral polyprotein processing, these proteases are suitable targets for the development of antiviral drugs. In order to obtain near-equipotent, broad-spectrum antivirals against alphacoronaviruses, betacoronaviruses, and enteroviruses, we pursued structure-based design of peptidomimetic α-ketoamides as inhibitors of main and 3C proteases. Six crystal structures of protease:inhibitor complexes were determined as part of this study. Compounds synthesized were tested against the recombinant proteases as well as in viral replicons and virus-infected cell cultures; most of them were not cell-toxic. Optimization of the P2 substituent of the α-ketoamides proved crucial for achieving near-equipotency against the three virus genera. The best near-equipotent inhibitors, 11u (P2 = cyclopentylmethyl) and 11r (P2 = cyclohexylmethyl), display low-micromolar EC50 values against enteroviruses, alphacoronaviruses, and betacoronaviruses in cell cultures. In Huh7 cells, 11r exhibits three-digit picomolar activity against Middle East Respiratory Syndrome coronavirus.3CLpro3C-like protease;3Cpro3C protease;A490absorbance at 490 nm;apantiperiplanar;BACbacterial artificial chromosome;CPEcytopathic effect;CVA16Coxsackievirus A16;CVB3Coxsackievirus B3;DMEMDulbecco’s modified minimal essential medium;EMEMEagle’s minimal essential medium;EVenterovirus;FIPVFeline Infectious Peritonitis Virus;FRETfluorescence resonance energy transfer;GlnLactamglutamine lactam;HCoVhuman coronavirus;HFMDHand, Foot, and Mouth Disease;HRVhuman rhinovirus;MERS-CoVMiddle-East respiratory syndrome coronavirus;Mpromain protease;Nsp5non-structural protein 5;NTRnon-translated region;OD498optical density at 498 nm;PEGpolyethylene glycol;RFUrelative fluorescence units;Rzribozyme;SARSsevere acute respiratory syndrome;SARS-CoVSARS coronavirus;- sc(-)-synclinal;SDstandard deviation;TLCthin-layer chromatography