Similarities and differences upon binding of naturally occurring Δ9-tetrahydrocannabinol-derivatives to cannabinoid CB1 and CB2 receptors

Abstract

We have here assessed, using Δ⁹-tetrahydrocannabinol (Δ⁹-THC) for comparison, the effect of Δ⁹-tetrahydrocannabinolic acid (Δ⁹-THCA) and of Δ⁹-tetrahydrocannabivarin (Δ9-THCV) that is mediated by human versions of CB₁, CB₂, and CB₁-CB₂ receptor functional units, expressed in a heterologous system. Binding to the CB₁ and CB₂ receptors was addressed in living cells by means of a homogeneous assay. A biphasic competition curve for the binding to the CB₂ receptor, was obtained for Δ⁹-THCV in cells expressing the two receptors. Signaling studies included cAMP level determination, activation of the mitogen-activated protein kinase pathway and ß-arrestin recruitment were performed. The signaling triggered by Δ⁹-THCA and Δ⁹-THCV via individual receptors or receptor heteromers disclosed differential bias, i.e. the bias observed using a given phytocannabinoid depended on the receptor (CB₁, CB₂ or CB₁-CB₂) and on the compound used as reference to calculate the bias factor (Δ⁹-THC, a selective agonist or a non-selective agonist). These results are consistent with different binding modes leading to differential functional selectivity depending on the agonist structure, and the state (monomeric or heteromeric) of the cannabinoid receptor. In addition, on studying Gi-coupling we showed that Δ⁹-THCV and Δ⁹-THCA and Δ⁹-THCV were able to revert the effect of a selective CB₂ receptor agonist, but only Δ9-THCV, and not Δ9-THCA, reverted the effect of arachidonyl-2'-chloroethylamide (ACEA 100 nM) a selective agonist of the CB₁ receptor. Overall, these results indicate that cannabinoids may have a variety of binding modes that results in qualitatively different effects depending on the signaling pathway that is engaged upon cannabinoid receptor activation.

Keywords: Bitopic drug; CB(1)-CB(2) receptor heteromers; Cannabis sativa L; Exosite; THCA; THCV.