Synthetic
cannabinoid receptor agonists (SCRAs) are an evolving
class of new psychoactive substances (NPS) with structurally diverse
compounds emerging each year. Due to the rapid pace at which these
drugs enter the market, there is often little or nil information regarding
the pharmacology of these substances despite widespread human use.
In this study, 12 recently emerged SCRAs (reported between 2018 and
2020) were synthesized, analytically characterized, and pharmacologically
evaluated using a live cell-based nanoluciferase complementation reporter
assay that monitors in vitro cannabinoid receptor
type 1 (CB1) activation via its interaction with β-arrestin
2 (βarr2). All synthesized SCRAs acted as agonists of CB1, although
differences in potency (EC50 = 2.33–5475 nM) and
efficacy (E
max = 37–378%) were
noted, and several structure–activity relationships were identified.
SCRAs featuring indazole cores (EC50 = 2.33–159
nM) were generally of equal or greater potency than indole analogues
(EC50 = 32.9–330 nM) or 7-azaindole derivatives
(EC50 = 64.0–5475 nM). Interestingly, with the exception
of APP-BINACA (E
max = 75.7%) and 5F-A-P7AICA
(E
max = 37.4%), all SCRAs showed greater
efficacy than the historical SCRA JWH-018 to which responses were
normalized (E
max = 142–378%). The
most potent CB1 agonists in the study were ADB-BINACA (EC50 = 6.36 nM), 4F-MDMB-BINACA (EC50 = 7.39 nM), and MDMB-4en-PINACA
(EC50 = 2.33 nM). Notably, all of these SCRAs featured
an indazole core as well as a “bulky” tert-butyl moiety in the pendant amino acid side chain. This study confirms
that recently detected SCRAs 4F-MDMB-BICA, 5F-MPP-PICA, MMB-4en-PICA,
CUMYL-CBMICA, ADB-BINACA, APP-BINACA, 4F-MDMB-BINACA, MDMB-4en-PINACA,
A-CHMINACA, 5F-AB-P7AICA, 5F-MDMB-P7AICA, and 5F-AP7AICA were all
able to activate the CB1 receptor in vitro, albeit to different extents,
and are potentially psychoactive in vivo. These results indicate that
further evaluation of these widely used NPS is warranted to better
understand the risks associated with human consumption of these drugs.