Table 1. Inhibition of Forskolin-Stimulated cAMP and βarrestin2 Recruitmenta.
| inhibition
of cAMP |
βarrestin2
recruitment |
|||
|---|---|---|---|---|
| compound(s) | EC50 (nM) (95% C.I.) | Emax (% CP55, 940) ± S.E.M | EC50 (nM) (95% C.I.) | Emax (% 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**** |
| FM-6b | 20 (10–150) | 76 ± 10*** | 63 (36–100)* | 45 ± 1.5**** |
| FD-22a | 73 (14–230) | 52 ± 3.4**** | >10,000 | 30 ± 1.9**** |
| FD-24a | 8.0 (5.1–13) | 38 ± 1.0**** | >10,000 | 43 ± 2.3**** |
| FD-25a | >10,000 | 21 ± 6.0**** | >10,000 | 46 ± 1.6**** |
| FD-27a | >10,000 | 22 ± 1.8**** | >10,000 | 41 ± 1.1**** |
| FD-32a | >10,000 | 21 ± 0.62**** | 560 (400–760) | 53 ± 1.4**** |
CB2R activity was quantified for cAMP inhibition using the DiscoveRx HitHunter assay (CHO hCB2R) in cells treated with compounds for 90 min and for βarrestin2 recruitment using the DiscoveRx PathHunter assay (CHO hCB2R) in cells treated with compounds for 90 min. Data were fit to a variable slope (three-parameter) nonlinear regression in GraphPad (v. 9). Data are mean with 95% confidence interval (C.I.) (EC50) or mean ± S.E.M, n = 6 independent experiments performed in triplicate. Statistical analyses were by nonoverlapping C.I. (EC50) or two-way ANOVA followed by Dunnett’s posthoc test (Emax, Table S1). *p < 0.05, ***p < 0.001, ****p < 0.0001 relative to CP55,940 within assay. Data from this Table is graphed in Figure 2. Statistical data for these graphs are presented in Table S1.