Inhibition of human recombinant T-type calcium channels by phytocannabinoids in vitro

T-type Ca channels (I_Ca ) regulate neuronal excitability and contribute to neurotransmitter release. The phytocannabinoids Δ⁹ -tetrahydrocannabinol and cannabidiol effectively modulate T-type I_Ca , but effects of other biologically active phytocannabinoids on these channels are unknown. We thus investigated the modulation of T-type I_Ca by low abundance phytocannabinoids.

A fluorometric (fluorescence imaging plate reader [FLIPR]) assay was used to investigate modulation of human T-type I_Ca (Ca_V 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.

In the FLIPR assay, all 11 phytocannabinoids tested modulated T-type I_Ca , with most inhibiting Ca_V 3.1 and Ca_V 3.2 more effectively than Ca_V 3.3. Cannabigerolic acid was the most potent inhibitor of Ca_V 3.1 (pIC₅₀ 6.1 ± 0.6) and Ca_V 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 Ca_V 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.

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 I_Ca 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 I_Ca modulators.