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: The comprehensive in vitro SAR in these cell-based assays for GAT211 congeners obtained from the fluorine-walk are given in , and those from nitrogen-walk are shown in . table ft1 table-wrap mode="anchored" t5 Table 2. caption a7 Compound Structure cAMP βarrestin2 cLogP Thermodynamic Aqueous solubility EC 50 (nM) E Max (%) EC 50 (nM) E Max (%) μM μg/mL 6d Open in a separate window 230 (140–370) 114 ± 6.8 940 (540–1,800) ^ 46 ± 9.5 § 5.03 7.8 2.7 9f Open in a separate window >10,000 81 ± 4.4 1,600 (1,300–2,000) 30 ± 1.1 § 4.16 11.2 3.8 9g Open in a separate window > 10,000 4.0 ± 2.5 ††† > 10,000 2.8 ± 2.4 3.91 23.6 8.1 9h Open in a separate window 450 (47–4,200) 5.2 ± 3.5 ††† > 10,000 5.1 ± 0.35 3.91 6.8 2.3 9i Open in a separate window 1,100 (1,100–1,700) * 39 ± 12 † >10,000 7.0 ± 0.89 4.16 159.0 54.6 9d Open in a separate window 1,100 (630–1,700) * 150 ± 12 >10,000 9.2 ± 8.7 § 3.91 4.9 1.7 9e Open in a separate window 3,100 (1,400–6,700) * 88 ± 18 1,600 (980–2,700) 19 ± 1.7 § 3.91 2.1 0.7 9a Open in a separate window 260 (140–480) 130 ± 7.8 >10,000 5.8 ± 3.8 § 4.15 2.8 1.0 9b Open in a separate window 3,100 (634–15,000) * 170 ± 39 >10,000 −8.1 ± 7.9 § 3.91 1.1 0.4 9c Open in a separate window 510 (330–780) 110 ± 6.4 4,000 (2,200–7,500) ^ 25 ± 4.4 § 3.91 36.7 12.6 9j Open in a separate window 1,900 (630–3,700) * 95 ± 16 >10,000 −1.4 ± 2.7 § 4.28 -- -- Open in a separate window PAM activity was quantified for cAMP inhibition using the DiscoveRx HitHunter assay (CHO hCB1R) in cells treated with CP55,940 at EC 20 + 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