We have discovered a family of synthetic oxazole-based macrocycles to be active against SARS-CoV-2. The
synthesis, pharmacological properties, and docking studies of the compounds are reported in this study. The
structure of the new macrocycles was confirmed by NMR spectroscopy and mass spectrometry. Compounds 13,
14, and 15a-c were evaluated for their anti-SARS-CoV-2 activity on SARS-COV-2 (NRC-03-nhCoV) virus in Vero-
E6 cells. Isopropyl triester 13 and triacid 14 demonstrated superior inhibitory activities against SARS-CoV-2
compared to carboxamides 15a-c. MTT cytotoxicity assays showed that the CC50 (50% cytotoxicity concentration) of 13, 14, and 15a-c ranged from 159.1 to 741.8 μM and their safety indices ranged from 2.50 to 39.1. Study of the viral inhibition via different mechanisms of action (viral adsorption, replication, or virucidal
property) showed that 14 had mild virucidal (60%) and inhibitory effects on virus adsorption (66%) at 20 μM
concentrations. Compound 13 displayed several inhibitory effects at three levels, but the potency of its action is
primarily virucidal. The inhibitory activity of compounds 13, 14, and 15a-c against the enzyme SARS-CoV-2
Mpro was evaluated. Isopropyl triester 13 had a significant inhibition activity against SARS-CoV-2 Mpro with
an IC50 of 2.58 μM. Large substituents on the macrocyclic template significantly reduced the inhibitory effects of
the compounds. Study of the docking of the compounds in the SARS CoV-2-Mpro active site showed that the most potent macrocycles 13 and 14 exhibited the best fit and highest affinity for the active site binding pocket. Taken together, the present study shows that the new macrocyclic compounds constitute a new family of SARS CoV-2-Mpro inhibitors that are worth being further optimized and developed.