Journal: Journal of medicinal chemistry

Article Title: Application of Fluorine- and Nitrogen-Walk Approaches: Defining the Structural and Functional Diversity of 2-Phenylindole Class of CB1 Receptor Positive Allosteric Modulators

doi: 10.1021/acs.jmedchem.9b01142

Figure Lengend Snippet: (a-c) CHO cells stably-expressing hCB1R were treated with 0.10 nM – 10 μM GAT compounds alone for 90 min and cAMP inhibition (a) or βarrestin2 recruitment was measured (b). Compound agonist bias between cAMP inhibition and βarrestin2 recruitment (ΔΔLogR (cAMP - βarr2)] is shown in c. (d-f) CHO cells stably-expressing hCB1R were treated with 100 nM CP55,940 + 0.10 nM – 10 μM GAT compounds for 90 min and cAMP inhibition (d) or βarrestin2 recruitment was measured (e). Compound PAM bias between cAMP inhibition and βarrestin2 recruitment (ΔΔLogR (cAMP - βarr2)] is shown in f. cAMP data are expressed as % Forskolin response within compound treatment. βarrestin2 recruitment data are expressed as % CP55,940 response. Data were fit to a nonlinear regression (4 parameter model, GraphPad v. 7) for statistics in Table 5; or fit to the operational model (eq. 1) to calculate bias (c,f). Data are mean ± S.E.M. or 95% CI (c,f), n ≥ 6 independent experiments performed in triplicate.

Article Snippet: 27 – 29 table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Compound Structure cAMP βarrestin2 cLogP EC 50 (nM) E Max (%) EC 50 (nM) E Max (%) 6d (GAT211) Open in a separate window 230 (140–370) 110 ± 6.8 940 (540–1,800) ^ 46 ± 9.5 § 5.03 6e Open in a separate window 130 (58–280) 120 ± 9.0 900 (730–1,100) ^ 37 ± 1.3 § 5.16 6f Open in a separate window 191 (96–380) 130 ± 8.5 740 (540–990) ^ 23 ± 1.2 § 5.16 6g Open in a separate window 110 (59–190) 120 ± 6.0 1,100 (880–1,300) ^ 38 ± 1.3 § 5.16 6h Open in a separate window 22 (8.4–58) * 120 ± 6.1 750 (630–880) ^ 45 ± 1.3 § 5.16 6i Open in a separate window 1400 (1,100–1,700) * 140 ± 6.2 710 (417–1,210) 7.0 ± 0.54 ††† , § 5.16 6j Open in a separate window 80 (29–220) 140 ± 13 † 1,300 (1,100–1,600) ^ 55 ± 2.0 § 5.16 6k Open in a separate window 1,600 (1,200–2,300) * 120 ± 6.2 >10,000 −2.2 ± 6.7 ††† , § 5.16 6a Open in a separate window 43 (28–66) * 110 ± 3.6 1,060 (840–1,300) ^ 63 ± 2.3 § 5.16 6b Open in a separate window 730 (280–1,900) 130 ± 18 2,500 (1,200–5,400) 19 ± 3.3 † , § 5.16 6c Open in a separate window 440 (280–700) 110 ± 6.6 >10,000 8.0 ± 2.7 ††† , § 5.16 6l Open in a separate window 58 (34–97) * 110 ± 5.0 1,140 (870–1,500) ^ 54 ± 2.8 § 5.3 6m Open in a separate window 21 (14–30) * 100 ± 3.2 680 (580–790) ^ 76 ± 1.8 ††† 5.3 6r Open in a separate window 28 (16–49) * 93 ± 4.2 750 (570–980) ^ 71 ± 3.3 5.43 6s Open in a separate window 9.1 (3.4–24) * 93 ± 4.9 510 (340–760) ^ 82 ± 4.8 †† 5.43 6n Open in a separate window 1,300 (330–6,000) 47 ± 5.6 ††† 1,100 (890–1,300) 32 ± 0.99 5.3 6o Open in a separate window 270 (190–380) 121 ± 5.4 700 (440–1,100) ^ 27 ± 1.9 § 5.3 6p Open in a separate window 1,150 (430–3,100) * 91 ± 12 2,100 (1,800–2,600) 56 ± 2.0 § 5.43 6q Open in a separate window 230 (160–340) 110 ± 5.3 1,100 (910–1,400) ^ 43 ± 1.6 § 5.43 Open in a separate window CB1 PAM activity was quantified for cAMP inhibition using the DiscoveRx HitHunter assay (CHO hCB1R) in cells treated with CP55,940 at EC20 + GAT211 analog for 30 min, and for βarrestin2 recruitment using the DiscoveRx PathHunter assay (CHO hCB1R) in cells treated with CP55,940 at EC20 + GAT211 analog for 90 min. Data were fit to a variable slope (4 parameter) non-linear regression in GraphPad (v. 7).

Techniques: Stable Transfection, Expressing, Inhibition