A Benchmark of Popular Free Energy Approaches Revealing the Inhibitors Binding to SARS-CoV2 Mpro

COVID-19 pandemic has killed millions of people worldwide since its outbreak in Dec 2019. The pandemic is caused by the SARS-CoV-2 virus whose main protease (Mpro) is a promising drug target since it plays a key role in viral proliferation and replication. Currently, designing an effective therapy is an urgent task, which requires accurately estimating ligand-binding free energy to the SARS-CoV-2 Mpro. However, it should be noted that the accuracy of a free energy method probably depends on the protein target. A highly accurate approach for some targets may fail to produce a reasonable correlation with experiment when a novel enzyme is considered as a drug target. Therefore, in this context, the ligand-binding affinity to SARS-CoV-2 Mpro was calculated via various approaches. The Autodock Vina (Vina) and Autodock4 (AD4) packages were manipulated to preliminary investigate the ligand-binding affinity and pose to the SARS-CoV-2 Mpro. The binding free energy was then refined using the fast pulling of ligand (FPL), linear interaction energy (LIE), molecular mechanics-Poission Boltzmann surface area (MM-PBSA), and free energy perturbation (FEP) methods. The benchmark results indicated that for docking calculations, Vina is more accurate than AD4 and for free energy methods, FEP is the most accurate followed by LIE, FPL and MM-PBSA (FEP > LIE > FPL > MM-PBSA). Moreover, the binding mechanism was also revealed by atomistic simulations. The vdW interaction is the dominant factor. The residues Thr25 , Thr26 , His41 , Ser46 , Asn142 , Gly143 , Cys145 , Glu166 , and Gln189 are essential elements affecting on the binding process. Furthermore, the Ser46 and related residues probably are important elements affecting the enlarge/dwindle of the SARS-CoV-2 Mpro binding cleft. The benchmark probably guide for further investigations using computational approaches.