5202
N. Shah, T. S. Scanlan / Bioorg. Med. Chem. Lett. 14 (2004) 5199–5203
synthetic glucocorticoid. However, our compounds have
comparable potencies to the natural glucocorticoid,
cortisol (data not shown). For both AP-1 and NF-jB
responses, the majorityof the nonsteroidal compounds
showed efficacies on the order of 50% or more of
DEX, suggesting all, save 27 have the potential to exert
significant anti-inflammatoryeffects in vivo.
References and notes
1. Lefstin, J. A.; Yamamoto, K. R. Nature 1998, 392, 885.
2. Davis, G. F. Clin. Dermatol. 1986, 4, 161.
3. Merkulova, T. I.; Merkulov, V. M.; Mitina, R. L. Mol.
Biol. (Mosk) 1997, 31, 714.
4. Reichardt, H. M.; Tuckermann, J. P.; Gottlicher, M.;
Vujic, M.; Weih, F.; Angel, P.; Herrlich, P.; Schutz, G.
EMBO J. 2001, 20, 7168.
Previous reports of nonsteroidal glucocorticoids demon-
strate dissociation in vitro,28–31 albeit between different
cell types. Ours is the first to examine dissociation in a
single cell type as shown in Table 2 (MMTV, AP-1,
NF-jB, and inset chart). The EC50 of DEX was divided
bythat of each compound for each of the three assays.
Comparison of the relative heights of the bars indicates
degree of dissociation of the three responses relative to
DEX. Following potency, this first panel of nonsteroidal
arylpyrazole ligands can be placed into four distinct
classes. Compounds 15–18 are slightlymore effective
in repressing NF-jB than AP-1 and GRE activation.
Compounds 19, 20, 23, 24, 28, and 29 show little disso-
ciative activity. 21 and 22 are equipotent with respect to
repression, and 5–7-fold selective for this activityover
transactivation. 25 and 26, however, are between 10–
14-fold selective for repression of an AP-1 response over
both GRE activation and NF-jB repression. Although
manyof the compounds tested were not as efficacious
as DEX in NF-jB repression while acting as full agon-
ists for activation, when compared to TAT activation
in CV-1 cells, our compounds are both more potent
and more efficacious in repressing AP-1 and NF-jB.
This cell specific difference of transactivation has been
reported for other gluocorticoids and is independent of
the composition of the synthetic GRE.32 Differences of
expression levels of co-regulatorytranscription factors
and of GR itself between the two cells types likely con-
tribute to the discrepancyof activation activityseen in
our assays.
5. Umland, S. P.; Schleimer, R. P.; Johnston, S. L. Pulm.
Pharmacol. Ther. 2002, 15, 35.
6. Hellal-Levy, C.; Couette, B.; Fagart, J.; Souque, A.;
Gomez-Sanchez, C.; Rafestin-Oblin, M. FEBS Lett. 1999,
464, 9.
7. Shah, N.; Scanlan, T. Abstracts of Papers of the American
Chemical Society 2001, 221, U46.
8. Paquette, L. A.; Wang, T. Z.; Philippo, C. M. G.; Wang,
S. P. J. Am. Chem. Soc. 1994, 116, 3367.
9. Invitrogen, Steroid Receptor Competitor AssayKits.
10. Iniguez-Lluhi, J. A.; Lou, D. Y.; Yamamoto, K. R. J.
Biol. Chem. 1997, 272, 4149.
11. Darimont, B. D.; Wagner, R. L.; Apriletti, J. W.; Stallcup,
M. R.; Kushner, P. J.; Baxter, J. D.; Fletterick, R. J.;
Yamamoto, K. R. Genes Dev. 1998, 12, 3343.
12. Spaete, R. R.; Mocarski, E. S. J. Virol. 1985, 56, 135.
13. Herrlich, P. Oncogene 2001, 20, 2465.
14. Wissink, S.; van Heerde, E. C.; Schmitz, M. L.; Kalkh-
oven, E.; van der Burg, B.; Baeuerle, P. A.; van der Saag,
P. T. J. Biol. Chem. 1997, 272, 22278.
15. Nissen, R. M.; Yamamoto, K. R. Genes Dev. 2000, 14,
2314.
16. Scanlan, T. S.; Shah, N. Preparation of Nonsteroidal
Ligands for the Glucocorticoid Receptor. U.S. Patent
20,030,176,478, Filed 22 January2002.
17. Ali, A.; Graham, D. W.; Quraishi, N.; Balkovec, J. M.;
Thompson, C. F. 1H-Benzo[f]indazol-5-yl Derivatives as
Selective Glucocorticoid Receptor Modulators. Patent
WO03086294, Filed 11 April 2002.
18. Hannah, J.; Kelly, K.; Patchett, A. A. J. Med. Chem. 1975,
18, 168.
19. Hirschmann, R. F.; Steinberg, N. G.; Buchschacher, P.;
Fried, J. H.; Kent, G. J.; Tishler, M.; Steelman, S. L. J.
Am. Chem. Soc. 1963, 85, 120.
These compounds are also GR specific (data not
shown). Binding to androgen receptor (AR) and proges-
terone receptor (PR) was measured with fluorescence
polarization assays.9 A radioligand whole cell binding
assaywas used to assess mineralocorticoid receptor
(MR) binding.33 In all cases selectivityfor GR binding
versus AR, PR, or MR is greater than two orders of
magnitude.
20. During preparation of this manuscript, a similar series of
compounds was reported byAli, A. et al. J. Med. Chem.
2004, 47, 2441.
21. Bui, T.; Barbas, C. F. Tetrahedron Lett. 2000, 41, 6951.
22. Hoyte, R. M.; Zhang, J. X.; Lerum, R.; Oluyemi, A.;
Persaud, P.; OꢁConnor, C.; Labaree, D. C.; Hochberg, R.
B. J. Med. Chem. 2002, 45, 5397.
23. Kumar, V.; Bell, M. R.; Wetzel, J. R.; Herrmann, J. L.;
McGarry, R.; Schane, H. P.; Winneker, R. C.; Snyder, B.
W.; Anzalone, A. J. J. Med. Chem. 1993, 36, 3278.
24. Zayia, G. H. Org. Lett. 1999, 1, 989.
25. Rogatsky, I.; Wang, J. C.; Derynck, M. K.; Nonaka, D.
F.; Khodabakhsh, D. B.; Haqq, C. M.; Darimont, B. D.;
Garabedian, M. J.; Yamamoto, K. R. Proc. Natl. Acad.
Sci. U.S.A. 2003, 100, 13845.
The in vitro selectivitywe have seen is on par with com-
pounds that have proven dissociation in an in vivo mod-
el.28,31 Clearly, with just this first panel of compounds, a
significant range of mechanism specific transcriptional
regulation has been observed, and it remains to be seen
whether these compounds prove to be as effective and
selective in vivo.
26. Rogatsky, I.; Waase, C. L.; Garabedian, M. J. J. Biol.
Chem. 1998, 273, 14315.
27. Starr, D. B.; Matsui, W.; Thomas, J. R.; Yamamoto, K.
R. Genes Dev. 1996, 10, 1271.
28. Coghlan, M. J.; Jacobson, P. B.; Lane, B.; Nakane, M.;
Lin, C. W.; Elmore, S. W.; Kym, P. R.; Luly, J. R.; Carter,
G. W.; Turner, R.; Tyree, C. M.; Hu, J.; Elgort, M.;
Rosen, J.; Miner, J. N. Mol. Endocrinol. 2003, 17,
860.
Acknowledgements
We are grateful to I. Rogatsky, B. Darimont, and
H. Luecke for plasmids. Research support was from
the National Institutes of Health (DK 57574).
29. Elmore, S. W.; Coghlan, M. J.; Anderson, D. D.; Pratt, J.
K.; Green, B. E.; Wang, A. X.; Stashko, M. A.; Lin, C.