Structure-Based Design and Evaluation of Peptidomimetic Inhibitors Against the Human Norovirus Protease

Norovirus is the leading cause of acute gastroenteritis worldwide and can lead to severe chronic infection in immunocompromised patients. Currently, there are no FDA approved therapeutics or vaccines against Norovirus infection. However, progress has been made in terms of identifying the Norovirus protease as a promising drug target due to its necessity in the production of mature viral proteins. Our study has focused on the development of targeted covalent inhibitors against the human strains of Norovirus protease by employing both computational and enzymatic design and evaluation techniques. Analysis of our recently solved apo and complexed crystal structures of Norovirus GI.1 and GII.4 proteases provided additional insights into the binding pocket and active site composition, which led to improved inhibitor efficacy. Potential compounds were designed and analyzed using Schrodinger (2019‐1) software, allowing for predictive ADME screening, covalent docking of inhibitors to the protein structure, and binding affinity analysis. The lead compounds were synthesized, then enzymatically screened against purified Norovirus protease using a FRET‐based assay, following in silico design and evaluation. To date, our lead compounds exhibit nanomolar activity against the Norovirus GII.4 protease as well as the less infectious GI.1 strain. Support or Funding Information Vera Fay Righthand Fellowship in Virology