Identification and biochemical analyses of selective CB₂ agonists

Cannabinoid CB₁ and CB₂ receptors are activated by Δ⁹-tetrahydrocannabinol, a psychoactive component of marijuana. The cannabinoid CB₁ receptor is primarily located in the brain and is responsible for the psychoactive side effects, whereas the cannabinoid CB₂ receptor is located in immune cells and is an attractive target for immune-related maladies. We identify small molecules that selectively bind to the cannabinoid CB₂ receptor and can be further developed into therapeutics. The affinity of three molecules, ABK5, ABK6, and ABK7, to the cannabinoid CB₂ receptor was determined with radioligand competition binding. The potency of G-protein coupling was determined with GTPγS binding. The three compounds bound selectively to the cannabinoid CB₂ receptor_, and no binding to the cannabinoid CB₁ receptor was detected up to 10 μM. Immunoblotting studies show that the amount of ERK1/2 and MEK phosphorylation increased in a G_i/o-dependent manner. Furthermore, an immune cell line (Jurkat cells) was treated with ABK5, and as a result, inhibited cell proliferation. These three compounds are novel cannabinoid CB₂ receptor agonists and hold promise to be further developed to treat inflammation and the often-associated pain.