Design of Drug‐Like Protein–Protein Interaction Stabilizers Guided By Chelation‐Controlled Bioactive Conformation Stabilization

Abstract

Protein-protein interactions( PPIs) of 14-3-3 proteins are am odel system for studyingPPI stabilization. The complex natural product Fusicoccin As tabilizes many 14-3-3 PPIs but is not amenable for use in SAR studies, motivating the search for more drug-like chemical matter.H owever, drug-like 14-3-3PPI stabilizers enablingsuch studies have remainede lusive. An X-ray crystal structure of aP PI in complex with an extremelyl ow potencys tabilizer uncovered an unexpected non-protein interacting, ligand-chelated Mg 2 + leading to the discovery of metal-ion-dependent 14-3-3 PPI stabilization potency.T his originates from an ovel chelationcontrolled bioactive conformation stabilization effect. Metal chelation hasb een associated with pan-assayi nterference compounds (PAINS) and frequent hitter behavior,b ut chelation can evidently also lead to true potencyg ains and find use as am edicinal chemistry strategy to guidec ompound optimization. To demonstrate this, we exploited the effect to designt he first potent, selective, and drug-like 14-3-3 PPI stabilizers.Supporting information and the ORCID identification number(s) for the author(s) of this article can be found under: https://doi.10 mm Mg 2 + ."Fold increase" (y-axis)i st he mean fold-increase of signal over baseline (no (R)-2). (e)Ca 2 + (red triangles) and Mn 2 + (purple squares)a lso increase apparente fficacy of (R)-2 in 14-3-3/ERa(pT 594 )FPa ssay.

Trisubstituted Pyrrolinones as Small-Molecule Inhibitors Disrupting the Protein–RNA Interaction of LIN28 and Let-7

“…This series of pyrrolinones and pyrazoles were previously reported as stabilizers of the protein–protein interaction (PPI) involving the plant 14-3-3 proteins, 33 , 36 , 37 which are small adapter proteins that interact with functionally diverse protein partners to play important regulatory roles in vital cellular events including signal transduction, cell cycle control, and apoptosis. 38 An analysis combining the reported PPI stabilizing activity between 14-3-3 and the plant plasma membrane H + -ATPase 2 (PMA2) and PRI inhibitory activity against LIN28– let-7 revealed in this study showed intriguing SAR information surrounding the pyrrolinones and pyrazoles ( Figure S7 ).…”

Discovering protein–protein interaction stabilisers by native mass spectrometry

“…To test this methodology further, two additional putative stabilisers were introduced: fusicoccin J (FC-J), a biosynthetic precursor to FC-A; 25 and pyrrolidone 1 (Pyr1), a weak stabiliser of several 14-3-3-partner interactions. 26,29,30 On their own, like with FC-A, FC-J binds more strongly to 14-3-3s-ERa complexes albeit to a lesser extent with only moderate stabilisation (S P ¼ 0.96, Fig. S13 †).…”

NMR Spectroscopy of supramolecular chemistry on protein surfaces

“…In recent studies, the reverse experiment, monitoring 15 N, 2 H-labeled 14-3-3ΔC protein in 1 H, 15 N-TROSY-HSQC experiments [ 103 ], was used to investigate the stabilizing effect of several compounds on the interaction of 14-3-3 with different unlabeled peptide epitopes derived from 14-3-3 cargo proteins [ 104 – 106 ]. E.g., a semi-synthetic ligand consisting of a fusicoccane diterpene core combined with different sugar moieties was designed to improve its properties as molecular glue for the 14-3-3/p53 complex [ 105 ].…”