biostudies-literatureIslam MSNIGMS NIH HHSU.S. Department of Health & Human Services | National Institutes of HealthNIH HHS350https://www.ebi.ac.uk/biostudies/studies/S-EPMC8763920biostudies-literatureUnknown13(1)We report the discovery of a facile peptide macrocyclization and stapling strategy based on a fluorine thiol displacement reaction (FTDR), which renders a class of peptide analogues with enhanced stability, affinity, cellular uptake, and inhibition of cancer cells. This approach enabled selective modification of the orthogonal fluoroacetamide side chains in unprotected peptides in the presence of intrinsic cysteines. The identified benzenedimethanethiol linker greatly promoted the alpha helicity of a variety of peptide substrates, as corroborated by molecular dynamics simulations. The cellular uptake of benzenedimethanethiol stapled peptides appeared to be universally enhanced compared to the classic ring-closing metathesis (RCM) stapled peptides. Pilot mechanism studies suggested that theNature communicationsUnprotected peptide macrocyclization and stapling via a fluorine-thiol displacement reaction.PMC8763920R01 GM116204R01GM116204R35GM22552R35 GM133468S10 OD020095R35GM133468R01 GM123296R01GM123296Kafley PKaneria KHLyu ZZhang SMaloney RVoelz VAIslam MSJunod SLYang WWang REGuan YZhao MBuuh ZYCohen CfalseUnprotected peptide macrocyclization and stapling via a fluorine-thiol displacement reaction.We report the discovery of a facile peptide macrocyclization and stapling strategy based on a fluorine thiol displacement reaction (FTDR), which renders a class of peptide analogues with enhanced stability, affinity, cellular uptake, and inhibition of cancer cells. This approach enabled selective modification of the orthogonal fluoroacetamide side chains in unprotected peptides in the presence of intrinsic cysteines. The identified benzenedimethanethiol linker greatly promoted the alpha helicity of a variety of peptide substrates, as corroborated by molecular dynamics simulations. The cellular uptake of benzenedimethanethiol stapled peptides appeared to be universally enhanced compared to the classic ring-closing metathesis (RCM) stapled peptides. Pilot mechanism studies suggested that the2022-01-01T00:00:00Z2022 Jan2026-05-07T22:20:45.74Z2025-02-19T03:53:30.886ZS-EPMC87639203503949010.1038/s41467-022-27995-5