Modulation of Recombinant Human T-Type Calcium Channels by Δ 9-Tetrahydrocannabinolic Acid In Vitro

Introduction: Low voltage-activated T-type calcium channels (T-type I_Ca), Ca_V3.1, Ca_V3.2, and Ca_V3.3, are opened by small depolarizations from the resting membrane potential in many cells and have been associated with neurological disorders, including absence epilepsy and pain. Δ⁹-tetrahydrocannabinol (THC) is the principal psychoactive compound in Cannabis and also directly modulates T-type I_Ca; however, there is no information about functional activity of most phytocannabinoids on T-type calcium channels, including Δ⁹-tetrahydrocannabinolic acid (THCA), the natural nonpsychoactive precursor of THC. The aim of this work was to characterize THCA effects on T-type calcium channels.

Materials and Methods: We used HEK293 Flp-In-TREx cells stably expressing Ca_V3.1, 3.2, or 3.3. Whole-cell patch clamp recordings were made to investigate cannabinoid modulation of I_Ca.

Results: THCA and THC inhibited the peak current amplitude Ca_V3.1 with pEC₅₀s of 6.0±0.7 and 5.6±0.4, respectively. THC (1 μM) or THC produced a significant negative shift in half activation and inactivation of Ca_V3.1, and both drugs prolonged Ca_V3.1 deactivation kinetics. THCA (10 μM) inhibited Ca_V3.2 by 53%±4%, and both THCA and THC produced a substantial negative shift in the voltage for half inactivation and modest negative shift in half activation of Ca_V3.2. THC prolonged the deactivation time of Ca_V3.2, while THCA did not. THCA inhibited the peak current of Ca_V3.3 by 43%±2% (10 μM) but did not notably affect Ca_V3.3 channel activation or inactivation; however, THC caused significant hyperpolarizing shift in Ca_V3.3 steady-state inactivation.

Discussion: THCA modulated T-type I_Ca currents in vitro, with significant modulation of kinetics and voltage dependence at low μM concentrations. This study suggests that THCA may have potential for therapeutic use in pain and epilepsy through T-type calcium channel modulation without the unwanted psychoactive effects associated with THC.