biostudies-literaturede Araujo ADNational Health and Medical Research CouncilAustralian Research Council895-904https://www.ebi.ac.uk/biostudies/studies/S-EPMC9257625biostudies-literatureUnknown3(7)Cyclic peptides that modulate protein-protein interactions can be valuable therapeutic candidates if they can be delivered intact to their target proteins in cells. Here we systematically compare the effects of different helix-inducing cyclization constraints on the capacity of a macrocyclic peptide component to confer α-helicity, protein-binding affinity, resistance to degradative proteases and cell uptake to a 12-residue peptide fragment of tumor suppressor protein p53. We varied the helix-inducing constraint (hydrocarbon, lactam, aliphatic or aromatic thioether, <i>etc.</i>) and the position of the cyclization linker (<i>i</i> to <i>i</i> + 4 or <i>i</i> to <i>i</i> + 7 bridges) in order to sculpt the macrocyclic size, stabilize its structure, and promote cell uptake. We find that rigidRSC chemical biologyLandscaping macrocyclic peptides: stapling hDM2-binding peptides for helicity, protein affinity, proteolytic stability and cell uptake.PMC9257625112890820095511145372DP1801032441117017DP160104442CE200100012de Araujo ADWu KCHoang HNLim JNguyen HTFairlie DPfalseLandscaping macrocyclic peptides: stapling hDM2-binding peptides for helicity, protein affinity, proteolytic stability and cell uptake.Cyclic peptides that modulate protein-protein interactions can be valuable therapeutic candidates if they can be delivered intact to their target proteins in cells. Here we systematically compare the effects of different helix-inducing cyclization constraints on the capacity of a macrocyclic peptide component to confer α-helicity, protein-binding affinity, resistance to degradative proteases and cell uptake to a 12-residue peptide fragment of tumor suppressor protein p53. We varied the helix-inducing constraint (hydrocarbon, lactam, aliphatic or aromatic thioether, <i>etc.</i>) and the position of the cyclization linker (<i>i</i> to <i>i</i> + 4 or <i>i</i> to <i>i</i> + 7 bridges) in order to sculpt the macrocyclic size, stabilize its structure, and promote cell uptake. We find that rigid2022-01-01T00:00:00Z2022 Jul2025-04-04T12:40:50.682Z2025-04-04T12:40:50.682ZS-EPMC92576253586617110.1039/d1cb00231g