Poxvirus targeted by RFdiffusion peptide-binders

Peptide-binders were generated by computational RFdiffusion to the F13L-homodimer interface sequences of poxviruses such as vaccinia and monkeypox. Compared to small-drug screenings and/or co-evolutions, peptide-binders have the advantages of higher affinities and easier chemical / recombinant synthesis. Among the hundreds of 20-30-mer peptide-binders randomly generated by RFdiffusion, some targeted vaccinia (VACV) F13L homodimer interface predicting low nanoMolar affinities. To improve its physiological stability, additional peptide sequences were computationally generated by cyclization/ hallucination. The resulting de novo cyclic peptide sequences predicted picoMolar affinities not only targeting the VACV F13L homodimer interfaces but also inner cavities previously identified by small-drug dockings to Tecovirimat-resistant monkeypox (MPXV) mutants. Because their targeting to numerous highly-conserved amino acids among poxviruses, and improved physiological stability against proteases, cyclic peptide-binders may be more adequate against resistant mutants of VACV and MPXV poxviruses.