In vitro μ-opioid receptor activation potential of U10 and β-U10, positional isomers of the synthetic opioid naphthyl U-47700

New synthetic opioids (NSOs) with diverse chemical structures continue to appear on recreational drug markets worldwide. U-type opioids have become one of the largest groups of non-fentanyl-related NSOs. Starting in 2020, a previously unreported U-compound coined "β-U10" (2-naphthyl U-47700; N-[2-(dimethylamino)cyclohexyl]-N-methylnaphthalene-2-carboxamide) was identified in Australia and the United States. β-U10 is a positional isomer of α-U10 (1-naphthyl U-47700), more commonly known as "U10." Here, the first comparative in vitro pharmacological characterization of naphthyl U-47700 (U10 and β-U10), together with the structural analogue U-47700 and fentanyl, is reported. Application of a cell-based μ-opioid receptor (MOR) activation (β-arrestin 2 recruitment) assay demonstrated β-U10 (EC₅₀ = 348 nM; E_max = 150% vs. hydromorphone) to be less potent than U-47700 (EC₅₀ = 116 nM; E_max = 154%) and fentanyl (EC₅₀ = 9.35 nM; E_max = 146%) but considerably more active than the α-isomer (EC₅₀ value in the μM range). For the latter, maximum receptor activation could not be reached at 100 μM. The difference in MOR activation potential for U10 and β-U10 stresses the importance of (analytical) differentiation between closely related analytes. The emergence of β-U10 on the recreational drug market is an example of the continuing emergence of non-fentanyl-related NSOs and further emphasizes the need to closely monitor fluctuations in the drug supply.