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B. W. Trotter et al. / Bioorg. Med. Chem. Lett. 21 (2011) 2354–2358
temperature for 15 min. Then, forskolin at EC70 was added and incubated at
Acknowledgments
room temperature for an additional 30 min. Detection of cAMP was performed
using Cisbio’s HRTF cAMP dynamic 2 kit following the manufacturer’s protocol
for cAMP detection. After adding d2-cAMP and anti-cAMP-cryptate to all the
wells, there was a final incubation at room temperature for 1 h. Plates were
then read on an EnVision plate reader (Perkin Elmer). CV values calculated for a
positive control compound (>250 replicates) were 59% and 70%, respectively,
for CB1 and CB2 IP values.
We would like to thank Joan S. Murphy and Dr. Charles W. Ross
III for HRMS analysis. We would also like to thank Janine N. Brouil-
lette and Dr. Steven M. Pitzenberger for NMR support.
11. Liu, X. R.; Smith, B. J.; Chen, C.; Callegari, E.; Becker, S. L.; Chen, X.; Cianfrogna,
J.; Doran, A. C.; Doran, S. D.; Gibbs, J. P.; Hosea, N.; Liu, J. H.; Nelson, F. R.; Szewc,
M. A.; Van Deusen, J. Drug Metab. Dispos. 2006, 34, 1443.
12. Nagakura, Y.; Okada, M.; Kohara, A.; Kiso, T.; Toya, T.; Iwai, A.; Wanibuchi, F.;
Yamaguchi, T. J. Pharmacol. Exp. Ther. 2003, 306, 490.
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13. GSK CB2 agonist clinical candidate GW-842166X is unique in that it is reported
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