A Novel Class of Norovirus Inhibitors Targeting the Viral Protease with Potent Antiviral Activity In Vitro and In Vivo
Jana Van Dycke,
Wenhao Dai,
Zoe Stylianidou,
Jian Li,
Arno Cuvry,
Emma Roux,
Bingqian Li,
Jasper Rymenants,
Lindsey Bervoets,
Peter de Witte,
Hong Liu,
Johan Neyts,
Joana Rocha-Pereira
Affiliations
Jana Van Dycke
Laboratory of Virology and Chemotherapy, KU Leuven–Department of Microbiology, Immunology and Transplantation, Rega Institute, 3000 Leuven, Belgium
Wenhao Dai
State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zu Chong Zhi Road, Shanghai 201203, China
Zoe Stylianidou
Laboratory of Virology and Chemotherapy, KU Leuven–Department of Microbiology, Immunology and Transplantation, Rega Institute, 3000 Leuven, Belgium
Jian Li
State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zu Chong Zhi Road, Shanghai 201203, China
Arno Cuvry
Laboratory of Virology and Chemotherapy, KU Leuven–Department of Microbiology, Immunology and Transplantation, Rega Institute, 3000 Leuven, Belgium
Emma Roux
Laboratory of Virology and Chemotherapy, KU Leuven–Department of Microbiology, Immunology and Transplantation, Rega Institute, 3000 Leuven, Belgium
Bingqian Li
State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zu Chong Zhi Road, Shanghai 201203, China
Jasper Rymenants
Laboratory of Virology and Chemotherapy, KU Leuven–Department of Microbiology, Immunology and Transplantation, Rega Institute, 3000 Leuven, Belgium
Lindsey Bervoets
Laboratory of Virology and Chemotherapy, KU Leuven–Department of Microbiology, Immunology and Transplantation, Rega Institute, 3000 Leuven, Belgium
Peter de Witte
Laboratory for Molecular Biodiscovery, KU Leuven–Department of Pharmaceutical and Pharmacological Sciences, 3000 Leuven, Belgium
Hong Liu
State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zu Chong Zhi Road, Shanghai 201203, China
Johan Neyts
Laboratory of Virology and Chemotherapy, KU Leuven–Department of Microbiology, Immunology and Transplantation, Rega Institute, 3000 Leuven, Belgium
Joana Rocha-Pereira
Laboratory of Virology and Chemotherapy, KU Leuven–Department of Microbiology, Immunology and Transplantation, Rega Institute, 3000 Leuven, Belgium
Human noroviruses (HuNoVs) are the most common cause of viral gastroenteritis resulting annually in ~219,000 deaths and a societal cost of ~USD 60 billion, and no antivirals or vaccines are available. Here, we assess the anti-norovirus activity of new peptidomimetic aldehydes related to the protease inhibitor rupintrivir. The early hit compound 4 inhibited the replication of murine norovirus (MNV) and the HuNoV GI.1 replicon in vitro (EC50 ~1 µM) and swiftly cleared the HuNoV GI.1 replicon from the cells. Compound 4 still inhibits the proteolytic activity. We selected a resistant GI.1 replicon, with a mutation (I109V) in a highly conserved region of the viral protease, conferring a low yield of resistance against compound 4 and rupintrivir. After testing new derivatives, compound 10d was the most potent (EC50 nanomolar range). Molecular docking indicated that the aldehyde group of compounds 4 and 10d bind with Cys139 in the HuNoV 3CL protease by a covalent linkage. Finally, compound 10d inhibited the replication of HuNoV GII.4 in infected zebrafish larvae, and PK studies in mice showed an adequate profile.
