TABLE 1.
Inhibition of forskolin-stimulated cAMP and βarrestin2 recruitment.
| Compound(s) | Inhibition of cAMP | βarrestin2 recruitment | ||
|---|---|---|---|---|
| EC50 (nM) (95% C.I.) | E max (% CP55,940) ± S.E.M | EC50 (nM) (95% C.I.) | E max (% CP55,940) ± S.E.M | |
| CP55,940 | 9.4 (3.4–29) | 100 ± 6.4 | 560 (410–760) | 100 ± 3.4 |
| EC-21a | >10,000 | 2.5 ± 0.53* | >10,000 | 1.4 ± 0.96* |
| LV62 | 58 (5.5–250) | 110 ± 5.3 | 63 (49–82)* | 65 ± 1.4* |
| 10 nM LV62 + EC21a | 4.3 (0.47–34) | 104 ± 5.2 | n.d | n.d |
| JR14a | 150 (51–220) | 41 ± 5.7* | 25 (16–39)* | 39 ± 0.93* |
| JR16a | 350 (78–850)* | 36 ± 8.6* | n.c | 27 ± 4.2* |
| JR22a | 62 (2.5–410) | 38 ± 5.5* | >10,000 | 33 ± 3.1* |
| JR26a | 110 (60–210)* | 41 ± 5.1* | >10,000 | 39 ± 6.1* |
| JR58a | 270 (36–440)* | 45 ± 2.9* | >10,000 | 51 ± 2.8* |
| JR60a | 770 (200–1,110)* | 46 ± 4.4* | >10,000 | 36 ± 9.2* |
| JR61a | 420 (14–630) | 46 ± 1.7* | >10,000 | 34 ± 2.2* |
| JR64a | 8.6 (5.9–12) | 38 ± 0.74* | >10,000 | 45 ± 2.4* |
hCB2R activity was quantified for cAMP inhibition using the DiscoveRx HitHunter® assay (CHO-K1 hCB2R) in cells treated with compounds for 90 min, and for βarrestin2 recruitment using the DiscoveRx PathHunter® assay (CHO-K1 hCB2R) in cells treated with compounds for 90 min. Data were fit to a variable slope (3 parameter) non-linear regression in GraphPad (v. 9.0). Data are mean with 95% confidence interval (C.I.) (EC50) or mean ± S.E.M. (E max), n = 6 independent experiments performed in triplicate. Statistical analyses were by non-overlapping C.I. or two-way ANOVA followed by Bonferroni’s post-hoc test. *p < 0.05 relative to CP55,940 within assay. n. d, not determined; n. c, not converged. E max values for data that were not fit to a non-linear regression are the mean from the maximum value observed.