Cathepsin-Targeting SARS-CoV-2 Inhibitors: Design, Synthesis, and Biological Activity.

Cathepsins (Cats) are proteases that mediate the successful entry of SARS-CoV-2 into host cells. We designed and synthesized a tailored series of 21 peptidomimetics and evaluated their inhibitory activity against human cathepsins L, B, and S. Structural diversity was realized by combinations of different C-terminal warhead functions and N-terminal capping groups, while a central Leu-Phe fragment was maintained. Several compounds were identified as promising cathepsin L and S inhibitors with values in the low nanomolar to subnanomolar range, for example, the peptide aldehydes and (, 2.67 nM, CatL; 0.455 nM, CatS; , 1.76 nM, CatL; 0.512 nM, CatS). The compounds' inhibitory activity against the main protease of SARS-CoV-2 (M) was additionally investigated. Based on the results at CatL, CatS, and M, selected inhibitors were subjected to investigations of their antiviral activity in cell-based assays. In particular, the peptide nitrile exhibited promising antiviral activity with an EC value of 38.4 nM in Calu-3 cells without showing cytotoxicity. High metabolic stability and favorable pharmacokinetic properties make suitable for further preclinical development.