P. Patel et al. / Bioorg. Med. Chem. Lett. 21 (2011) 1987–1990
1989
Meakins-Christie Laboratories–MUHC-RI are supported in part by
a Center grant from Le Fonds de la Recherche en Santé du Québec
as well as by the J. T. Costello Memorial Research Fund. J.R. also
wishes to acknowledge the National Science Foundation for the
AMX-360 (Grant Number CHE-90-13145) and Bruker 400 MHz
(Grant Number CHE-03-42251) NMR instruments. The content is
solely the responsibility of the authors and does not necessarily
represent the official views of the National Heart, Lung, And Blood
Institute or the National Institutes of Health.
4
3
2
1
3
2
1
Dig
5oETE
Veh
5oETE
Dig
F3-5o
A. Supplementary data
Supplementary data associated with this article can be found, in
0
1
2
3
4
Time (min)
Figure 1. Effects of 5-oxo-ETE and trifluoro-5-oxo-ETE on intracellular calcium
levels in indo-1-labeled neutrophils. Either vehicle or trifluoro-5-oxo-ETE (100 nM)
was added after 1 min, followed by 5-oxo-ETE (10 nM) at 2.5 min and digitonin
(0.1%) at 3.5 min. Fluorescence due to the binding of Ca++ to indo-1 was monitored
using excitation and emission wavelengths of 331 and 410 nm.
References and notes
1. Rokach, J. Leukotrienes and Lipoxygenases, Chemical, Biological and Clinical
Aspects, 1st ed.; Elsevier Science: Amsterdam The Netherlands, 1989. Vol. 11.
2. Atkinson, J. G.; Rokach, J. Synthesis of the Leukotrienes and Other
Lipoxygenase-Derived Products In CRC Handbook of Eicosanoids:
Prostaglandins and Related Lipids. Chemical and Biochemical Aspects, Part B;
Willis, A. L., Ed.; CRC Press: Boca Raton, FL, 1987; 1, p 175.
3. Vane, J.; O’Grady, J. Therapeutic Applications of Prostaglandins; Edward Arnold:
London, 1993. 277.
4. Nicolaou, K. C.; Petasis, N. A. Synthesis of Eicosanoids In CRC Handbook of
Eicosanoids: Prostaglandins and Related Lipids. Chemical and Biochemical Aspects,
Part B; Willis, A. L., Ed.; CRC Press: Boca Raton, FL, 1987; 1, p 1.
5. Spokas, E. G.; Rokach, J.; Wong, P. Y.-K. Leukotrienes, Lipoxins, and
Hydroxyeicosatetraenoic acids In Methods in Molecular Biology; Lianos, E. A.,
Ed.; Humana Press: Totowa, NJ, 1999; p 213.
6. Lau, C. K.; Adams, J.; Guindon, Y.; Rokach, J. Chemistry of the Leukotrienes and
Other Lipoxygenase Products. In Leukotrienes and Lipoxygenase: Chemical,
Biological, and Clinical Aspects; Rokach, J., Ed.; Elsevier Press: Amsterdam,
1989; p 1. Chapter 1.
7. Rokach, J.; Guindon, Y.; Young, R. N.; Adams, J.; Atkinson, J. G. Synthesis of the
Leukotrienes In The Total Synthesis of Natural Products; ApSimon, J. W., Ed.;
Wiley-Interscience: New York, 1988; 7,.
8. Rokach, J.; Adams, J. Acc. Chem. Res. 1985, 19, 87.
9. Adams, J.; Fitzsimmons, B. J.; Girard, Y.; Leblanc, Y.; Evans, J. F.; Rokach, J. J. Am.
Chem. Soc. 1985, 107, 464.
10. Morrow, J. D.; Hill, K. E.; Burk, R. F.; Nammour, T. M.; Badr, K. F.; Roberts, L. J., II
Proc. Natl. Acad. Sci. U.S.A. 1990, 87, 9383.
11. Hwang, S. W.; Adiyaman, M.; Khanapure, S.; Schio, L.; Rokach, J. J. Am. Chem.
Soc. 1994, 116, 10829.
12. Praticò, D.; Barry, O. P.; Lawson, J. A.; Adiyaman, M.; Hwang, S. W.; Khanapure,
S. P.; Iuliano, L.; Rokach, J.; FitzGerald, G. A. Proc. Natl. Acad. Sci. U.S.A. 1998, 95,
3449.
5-oxo-ETE, as illustrated by Figure 1. The IC50 for 5-oxo-ETE-
induced desensitization was 3 nM, whereas that for C20-trifluoro-
5-oxo-ETE was 7.5 nM (Fig. 2B).
Although C20-trifluoro-5-oxo-ETE is some what less potent an
agonist than 5-oxo-ETE, it is considerably more potent than the
5-oxo-20-HETE 7, a C20-hydroxy metabolite of 5-oxo-ETE, indicat-
ing that addition of a substituent at this position does not necessar-
ily dramatically reduce biological activity. The potent inhibitory
effect of C20-trifluoro-5-oxo-ETE on 5-oxo-ETE-induced calcium
mobilization (Fig. 2B) suggests that the trifluoro derivative binds
strongly to the 5-oxo-ETE receptor, whereas the limited maximal
response suggests that it is a partial agonist, having some antago-
nist properties. These results suggest that further modification of
the C20 methyl group may lead to the identification of 5-oxo-ETE
analogs with potent agonist or antagonist effects. Also, Figure 2B
suggests that an appropriately radio-labeled C20-trifluoro-5-oxo-
ETE could be useful in generating an OXE-R binding assay and for
long term in vivo experimentation.
Acknowledgements
13. Zamboni, R.; Rokach, J. Tetrahedron Lett. 1983, 24, 999.
14. Corey, E. J.; Albright, J. O.; Barton, A. E.; Hashimoto, S. J. Am. Chem. Soc. 1980,
102, 1435.
15. Dussault, P.; Lee, I. J. Org. Chem. 1995, 60, 218.
16. Murphy, R. C.; Hammarstrom, S.; Samuelsson, B. Proc. Natl. Acad. Sci. U.S.A 1979,
76, 4275.
This work was supported by grants from the National Heart,
Lung, And Blood Institute (J.R.; Award Number R01HL081873),
the Canadian Institutes of Health Research (W.S.P.; MOP-6254),
and the Quebec Heart and Stroke Foundation (W.S.P.). The
100
100
A
B
5o-ETE
75
75
F3-5o
5o-ETE
50
50
25
0
F3-5o
25
0
0.1
1
10 100 1000
0.1
1
10 100 1000
Concentration (M)
Concentration (M)
Figure 2. Concentration-response relationships for the effects of 5-oxo-ETE and trifluoro-5-oxo-ETE on intracellular calcium levels in neutrophils. (A) Agonist effects of 5-
oxo-ETE and trifluoro-5-oxo-ETE on intracellular Ca++ levels in neutrophils. (B) Inhibition of 5-oxo-ETE (10 nM)-induced Ca++ mobilization in neutrophils due to prior
treatment with different concentrations of 5-oxo-ETE or trifluoro-5-oxo-ETE as shown in Figure 1.