Modified cannabinoids unlock dual pain-channel activation

TRPA1 and TRPV1 are nociceptive ion channels that integrate noxious chemical stimuli and are attractive targets for non-opioid analgesics. The phytocannabinoids cannabidiol (CBD) and cannabigerol (CBG) modulate both channels, but their pharmacological profiles and underlying structural determinants remain poorly defined. Here, we used rational ligand design to convert CBD and CBG into potent dual TRPA1/TRPV1 agonists and to probe a conserved lipid-sensing architecture in both channels. Guided by the structural features of the TRPV1 vanilloid-binding site and our previously developed TRPA1 cannabinoid-binding site model, we incorporated an aryl carbamate anchor into the CBD and CBG scaffolds. We compared their activity with that of the parent phytocannabinoids in heterologous expression systems using calcium imaging and whole-cell patch-clamp recordings. Whereas CBD and CBG preferentially activated TRPA1 and were weak TRPV1 agonists at physiologically relevant concentrations, carbamate functionalization markedly increased potency and efficacy at TRPV1 while preserving robust TRPA1 activation. In contrast to the parent phytocannabinoids, carbamate derivatives did not potentiate TRPV2 responses, demonstrating channel selectivity. Site-directed mutagenesis and molecular docking localized carbamate-based ligand binding to the canonical vanilloid binding site in TRPV1 and the non-electrophilic cannabinoid-binding site in TRPA1, consistent with engagement of a shared lipid-sensing pocket. Carbamate derivatives also activated nociceptive neurons in primary rat trigeminal ganglion cultures in a TRPV1-dependent manner, as confirmed by pharmacological block with the selective TRPV1 antagonist BCTC. These findings define a conserved structural framework for lipid ligand recognition in TRPA1 and TRPV1 and provide a rational starting point for developing dual-target modulators of peripheral nociceptors.