Transient receptor potential melastatin member 8 (TRPM8) is a polymodally activated ion channel that serves as the primary human cold sensor and a validated therapeutic target for various indications. Nevertheless, TRPM8-targeting compounds often fail clinically due to on-target side effects related to thermosensation and thermoregulation. Here, we demonstrate that TRPM8 protein dynamics, as detected by solution NMR, serve as a quantifiable and predictive intermediate linking ligand chemical structure to cellular function. Using a rationally designed library of TRPM8-regulating small molecules, cheminformatic clustering showed that chemical structure correlates with agonist or antagonist activity, which was validated by automated patch-clamp electrophysiology, showing that EC 50 /IC 50 values align with cheminformatic predictions. Alternatively, cryo-EM structural analysis of TRPM8 shows limited conformational differences across ligand-bound states, suggesting that static structures alone do not fully explain TRPM8 functional outcomes. To probe dynamic mechanisms, we engineered a 19 F-labeled human TRPM8 voltage-sensing-like domain (VSLD) construct and applied NMR spectroscopy. The resulting NMR studies show that ligand binding conformationally selects distinct dynamic ensembles, with spectral signatures correlating to both chemical structure and cellular potency. These NMR-detected dynamics signatures were robust across chemotype diversity and experimental conditions. Our findings support a model of conformational selection in TRPM8, where ligand-induced redistribution of dynamic states governs functional output. This dynamic bridge between structure and function potentially offers a predictive framework for TRPM8 drug discovery and other dynamic systems, where static structures alone may be insufficient. The integration of NMR-based dynamics, cheminformatics, and cellular assays may provide a distinct platform for rational therapeutic design in complex biological systems.
Horn et al. (Sun,) studied this question.