Identification of hACE2-interacting sites in SARS-CoV-2 spike receptor binding domain for antiviral drugs screening

•Peptide 391–465 aa is the major hACE2-interacting sites in SARS-CoV-2 spike RBD.•Essential amino acid residues (403R, 449Y, 454R) are critical for the interaction between the SARS-CoV-2 spike RBD and hACE2.•Spike 454R, 449Y, 403R, 439N and 440N are critical for infectivity of SARS-CoV-2.•Cephalosporin derivatives efficiently block infection of the pseudoviruses with wild type spike or new variants in vitro.•Cefixime effectively inhibited live SARS-CoV-2 infection in vitro.