Br J Pharmacol. 2022 Mar 27. doi: 10.1111/bph.15842. Online ahead of print.
BACKGROUND AND PURPOSE: T-type Ca channels (ICa ) regulate neuronal excitability and contribute to neurotransmitter release. The phytocannabinoids Δ9 -tetrahydrocannabinol and cannabidiol effectively modulate T-type ICa , but effects of other biologically active phytocannabinoids on these channels are unknown. We thus investigated the modulation of T-type ICa by low abundance phytocannabinoids.
EXPERIMENTAL APPROACH: A fluorometric (FLIPR) assay was used to investigate modulation of human T-type ICa (CaV 3.1, 3.2 and 3.3) stably expressed in FlpIn-TREx HEK293 cells. The biophysical effects of some compounds were examined using whole-cell patch clamp recordings from the same cells.
KEY RESULTS: In the FLIPR assay, all eleven phytocannabinoids tested modulated T-type ICa , with most inhibiting CaV 3.1 and CaV 3.2 more effectively than CaV 3.3. Cannabigerolic acid was the most potent inhibitor of CaV 3.1 (pIC50 6.1 ± 0.6) and CaV 3.2 (pIC50 6.4 ± 0.4); in all cases phytocannabinoid acids were more potent than their corresponding neutral forms. In patch clamp recordings, cannabigerolic acid inhibited CaV 3.1 and 3.2 with similar potency to the FLIPR assay, the inhibition was associated with significant hyperpolarizing shift in activation and steady state inactivation of these channels. In contrast, cannabidiol, cannabidivarin and cannabigerol only affected channel inactivation.
CONCLUSION AND IMPLICATIONS: Modulation of T-type calcium channels is a common property of phytocannabinoids, which all increase steady state inactivation at physiological membrane potentials, with some also affecting channel activation. Thus, T-type ICa may be a common site of action for phytocannabinoids, and the diverse actions of phytocannabinoids on channel gating may provide insight into structural requirement for selective T-type ICa modulators.