Feature-Based Molecular Networking for the Targeted Identification of G_q-Inhibiting FR900359 Derivatives

Both the soil bacterium Chromobacterium vaccinii and the bacterial endosymbiont Candidatus Burkholderia crenata of the plant Ardisia crenata are producers of FR900359 (FR). This cyclic depsipeptide is a potent and selective G_q protein inhibitor used extensively to investigate the intracellular signaling of G protein coupled receptors (GPCRs). In this study, the metabolomes of both FR producers were investigated and compared using feature-based molecular networking (FBMN). As a result, 30 previously unknown FR derivatives were identified, one-third being unique to C. vaccinii. Guided by MS, a novel FR derivative, FR-6 (compound 1), was isolated, and its structure unambiguously established. In a whole-cell biosensing assay based on detection of dynamic mass redistribution (DMR) as readout for G_q inhibition, FR-6 suppressed G_q signaling with micromolar potency (pIC₅₀ = 5.56). This functional activity was confirmed in radioligand binding assays (pK_i = 7.50). This work demonstrates the power of molecular networking, guiding the way to a novel G_q-inhibiting FR derivative and underlining the potency of FR as a G_q inhibitor.