Development of a GC-376 Based Peptidomimetic PROTAC as a Degrader of 3‑Chymotrypsin-like Protease of SARS-CoV‑2

We have applied a proteolysis targeting chimera (PROTAC) technology to obtain a peptidomimetic molecule able to trigger the degradation of SARS-CoV-2 3-chymotrypsin-like protease (3CL^Pro). The PROTAC molecule was designed by conjugating a GC-376 based dipeptidyl 3CL^Pro ligand to a pomalidomide moiety through a piperazine–piperidine linker. NMR and crystallographic data complemented with enzymatic and cellular studies showed that (i) the dipeptidyl moiety of PROTAC binds to the active site of the dimeric state of SARS-CoV-2 3CL^Pro forming a reversible covalent bond with the sulfur atom of catalytic Cys145, (ii) the linker and the pomalidomide cereblon-ligand of PROTAC protrude from the protein, displaying a high degree of flexibility and no interactions with other regions of the protein, and (iii) PROTAC reduces the protein levels of SARS-CoV-2 3CL^Pro in cultured cells. This study paves the way for the future applicability of peptidomimetic PROTACs to tackle 3CL^Pro-dependent viral infections.