A Novel Class of Norovirus Inhibitors Targeting the Viral Protease with Potent Antiviral Activity In Vitro and In Vivo

Human noroviruses (HuNoVs) are the most common cause of viral gastroenteritis resulting annually in similar to 219,000 deaths and a societal cost of similar to 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 similar to 1 mu M) 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.

Automated summary

New peptidomimetic aldehydes have shown anti-norovirus activity, with compound 10d being the most potent. Molecular docking revealed that the aldehyde group of these compounds binds with the HuNoV 3CL protease via a covalent linkage. The study also demonstrated the effectiveness of these compounds in inhibiting different types of noroviruses in mouse and zebrafish models.