5332
A. B. Pinkerton et al. / Bioorg. Med. Chem. Lett. 14 (2004) 5329–5332
OH
O
OH
O
R1
R1
Br
R2
a
+
NC
b,c
HO
30
HO
28
29
O
OH
R1
O
R2
O
N
4, 14-22
N
N
HN
Scheme 1. Reagents and conditions: (a) R2COCl, AlCl3, CH2Cl2; (b) K2CO3, acetone, 45°C; (c) TMS-N3, Bu2SnO, toluene, reflux.
13. (a) Johnson, M. P.; Baez, M.; Jagdmann, G. E., Jr.;
Britton, T. C.; Large, T. H.; Callagaro, D. O.; Tizzano, J.
assays, displayed a potency of 1.7lM with 52% potenti-
ation.14 Further exploration of this series of compounds
is in progress and will be disclosed in due course.
P.; Monn, J. A.; Schoepp, D. D. J. Med. Chem. 2003, 46,
3189; (b) Barda, D. A.; Wang, Z.; Britton, T. C.; Henry, S.
S.; Jagdmann, G. E.; Coleman, D. S.; Johnson, M. P.;
References and notes
Andis, S. L.; Schoepp, D. D. Bioorg. Med. Chem. Lett.
2004, 14, 3099.
14. Schaffhauser, H.; Rowe, B. A.; Morales; Chavez-Noriega,
L. E.; Yin, R.; Jachec, C.; Rao, S. P.; Bain, G.; Pinkerton,
A. B.; Vernier, J.-M.; Bristow, L. J.; Varney, M. A.;
Daggett, L. P. Mol. Pharmacol. 2003, 64, 798–810.
15. Screening of compounds was carried out using a Ca2+ flux
functional (FLIPR384) assay using a stable cell line co-
expressing the human mGlu2 receptors coupled to a
promiscuous G-protein (Ga16). Receptor activity was
detected by changes in [Ca2+], measured using the
fluorescent, Ca2+ sensitive dye fura-2. For further infor-
mation, see: Varney, M. A.; Cosford, N. D.; Jachec, C.;
Rao, S. P.; Sacaan, A.; Lin, F. F.; Bleicher, L.; Santori, E.
M.; Flor, P. J.; Allgeier, H.; Gasparini, F.; Kuhn, R.;
Hess, S. D.; Velicelebi, G.; Johnson, E. C. J. Pharmacol.
Exp. Ther. 1999, 290, 170–181.
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Screening hits were confirmed and further characterized in
the [35S]-GTPcS binding assay using a cell line expressing
human mGlu2 receptor. See Ref. 14 for a detailed
description of this assay. Compounds displayed a similar
profile and potency in both assays.
16. Pinkerton, A. B.; Vernier, J.-M.; Schaffahuser, H.; Rowe,
B. A.; Campbell, U. C.; Rodriguez, D. E.; Lorrain, D. S.;
Baccei, C. S.; Daggett, L. P.; Bristow, L. J. J. Med. Chem.
2004, ASAP articles.
17. For the GTPcS assay, an EC10 (1lM) of glutamate was
added to the cell line followed immediately by the test
compound at varying concentrations. The response was
then compared to a response using a saturating amount of
glutamate (1mM) to give both an EC50 and a percent
potentiation (the response normalized to the maximum
response of glutamate alone). The same experiment was
carried out in the absence of glutamate to test if the
compound was truly a positive allosteric modulator. Non-
specific binding was determined by addition of 10lM
unlabeled GTPcS.
18. For some similar reactions, see: Sprenger, R. D.; Ruoff, P.
M.; Frazer, A. H. J. Am. Chem. Soc. 1950, 72, 2874–2876;
Ahluwalia, V. K.; Singh, R. P.; Tripathi, R. P. Gazz.
Chim. Ital. 1984, 114(7–8), 359–361.
19. All final compounds displayed spectral data (NMR, MS)
that was consistent with the assigned structure.
11. (a) Grillon, C.; Cordova, J.; Levine, L. R.; Morgan, C. A.,
III Psychopharmacology 2003, 168, 446–454; (b) Schoepp,
D. D.; Wright, R. A.; Levine, L. R.; Gaydos, B.; Potter,
W. Z. Stress 2003, 6, 189–197.
12. Knoflach, F.; Mutel, V.; Jolidon, S.; Kew, J. N. C.;
Malherbe, P.; Viera, E.; Wichmann, J.; Kemp, J. A. Proc.
Natl. Acad. Sci. U.S.A. 2001, 98, 13402–13407.