Please find below a summary of the scientific and technological achievement, ordered along the two main objectives and 4 sub-objectives as described in Annex 1.
Objective 1 – Mechanistic understanding of PPI stabilization between the 14-3-3 scaffold protein and IDPs
Our work established how small molecules can stabilize native 14-3-3 interactions with intrinsically disordered proteins (IDPs) to create functional ternary complexes.
Objective 1A – From peptide to protein – Implications on conformational selection and multivalency
We then progressed from minimal peptide epitopes toward full-length proteins, explicitly testing how multivalency and conformational selection shape stabilization. In Tau, a paradigmatic IDP with multiple phosphor-dependent 14-3-3 binding modes, reversible covalent imine tethers were engineered as molecular glues that selectively stabilize the pS214 site over other Tau sites (e.g. pS324), demonstrating that site topology and composite interfaces can be exploited for precision selectivity even within the same disordered partner (e.g. RSC Med. Chem. 16, 2190–2201 (2025)). Complementary proximity-enhanced Cys-His crosslinking methodology provided structural constraints in otherwise dynamic complexes, showing that ternary complex formation and cooperativity can be captured in solution and read out across IDPs. Together this has advanced our ability to relate epitope-level binding to protein-level recognition (e.g. Chem. Sci. 16, 3523–3535 (2025)).
Objective 1B – Selectivity via cooperativity – A universal concept for diverse PPI stabilization
We consolidated cooperativity driven selectivity as a universal principle across diverse 14-3-3 clients. A scaffold hopping strategy using Groebke–Blackburn–Bienaymé multi-component reaction (MCR) chemistry produced new drug like molecular glues for the 14-3-3/ERα complex, with multiple ternary crystal structures and deeply rooted in strong biophysical analysis. These data quantified cooperativity and showed that glue potency arises from synergistic binding at the composite interface, not just binary affinity to either partner (e.g. Nat. Commun. 16, 6467 (2025)). Mechanistic insights from the ChREBPα study further generalized the concept: enhancing native client sequestration by 14-3-3 under stress yields functional selectivity at the cellular level, illustrating how cooperativity translates into phenotypic control for otherwise “undruggable” transcription factors (e.g. Nat. Commun. 16, 2110 (2025)).
Objective 2 – PPI stabilization approaches to drug the undruggable; biomolecular condensates and IDPs
Objective 2A – Biomolecular condensates – The next frontier in PPI stabilization
Building on these platforms, we engineered condensate interfaces via dynamic protein insertion, enhancing droplet stability and compartmental control for sustained PPI modulation, which is key to deploying molecular glues in realistic mesoscale environments (e.g. J. Am. Chem. Soc. 147, 18412 (2025)). In parallel, light-activatable protease logic (LaTEV) inside coacervates delivered spatiotemporal release of protein cargos, establishing stimulus-responsive sender-receiver architectures and demonstrating how condensates can host programmable biochemical control applicable to ternary complex formation and dissolution (e.g. Adv. Biol. 9, 2400353 (2024)).
Objective 2B – Mechanism and data-driven small-molecule PPI stabilization – A case study on 14-3-3–Tau
For Objective 2B, we unified mechanistic, structural, and data-driven elements in a focused case study on 14-3-3/Tau (e.g. Commun. Biol. 8, 1139 (2025)). The selective, reversible covalent glues for Tau pS214 showcased site-level precision across Tau’s multivalent landscape; biophysical assays and co-crystal structures revealed how composite interfaces and ternary cooperativity yield stabilization without off-target engagement of other Tau sites (e.g. RSC Med. Chem. 16, 2190 (2025)). The Deep learning predictor guided epitope selection and motif discovery for 14-3-3 clients, informing glue design iterations and wet-lab validation that linked sequence features to cooperativity outcomes (e.g. Digital Discovery 4, 2602–2614 (2025)).
