L. Mugherli et al. / Bioorg. Med. Chem. Lett. 16 (2006) 4488–4491
4491
10. Fersht, A. In Enzyme Structure and Mechanism; W.H.
Freeman & Co: New York, 1985.
display low absorption within the fluorescein excitation/
emission range and will not interfere with fluorescence
measurements.29 Inhibitor assay could easily be per-
formed in microtiter plates provided that purified en-
zymes with a high enough cleavage efficiency are used.
11. (a) Breeuwer, P.; Drocourt, J. L.; Bunschoten, N.;
Zwietering, M. H.; Rombouts, F. M.; Abee, T. Appl.
Environ. Microbiol. 1995, 611, 614–619; (b) Lomolino,
G.; Lante, A.; Crapisi, A.; Spettoli, P.; Curioni, A.
Electrophoresis 2001, 22, 1021–1023; (c) Kierat, R. M.;
Kraemer, R. . Bioorg. Med. Chem. Lett. 2005, 15, 4824–
4827.
12. (a) Caturla, F.; Enjo, J.; Bernabeu, C. M.; Le Serre, S.
Tetrahedron 2004, 60, 1903–1911; (b) Takakusa, H.;
Kikuchi, K.; Urano, Y.; Sakamoto, S.; Yamaguchi, K.;
Nagano, T. J. Am. Chem. Soc. 2002, 124, 1653–1657.
13. (a) Qingping, W.; Scheigetz, J.; Roy, B.; Ramachandran,
C.; Gresser, M. J. Biochim. Biophys. Acta 2002, 1601, 19;
(b) Gee, K. R. Bioorg. Med. Chem. Lett. 1999, 9, 1395.
14. (a) Melhado, L. L.; Stuart, P. W.; Steven, L. P.; Mangel,
W. F. J. Am. Chem. Soc. 1982, 104, 7299; (b) Thomas, J.
J.; Eveland, W. C.; Medzon, E. L.; Christian, W.; Wylie,
D. E.; Burckhalter, J. H.; Jones, R. H. Proc. Soc. Exp.
Biol. Med. 1972, 140, 179.
In conclusion, new fluorogenic MFE-ester substrates
have been synthesized in a fast and convenient way.
The substrates may be used as sensitive indicators of a
wide range of proteolytic activities, and show promise
for screening libraries of protease inhibitors.
Acknowledgment
This work was supported by a grant from the Associa-
tion pour la Recherche sur le Cancer to M.Y.B.
Supplementary data
15. Zaikova, T. O.; Rukavishnikov, A. V.; Birrell, G. B.;
Griffith, O. H.; Keana, J. F. Bioconjug. Chem. 2001, 12,
307.
16. Kirschke, H.; Barrett, A. J. In Lysosomes: Their Role in
Protein Breakdown; Glaumann, H., Ballard, F. J., Eds.;
Academic Press: London, 1987; p 193.
17. (a) Mikkelsen, T.; Yan, P. S.; Ho, K. L.; Sameni, M.;
Sloane, B. F.; Rosenblum, M. L. J. Neurosurg. 1995, 83,
285; (b) Sloane, B. F.; Dunn, J. R.; Honn, K. V. Science
1981, 212, 1151.
Supporting information contains experimental informa-
tion and characterization data for all new compounds.
Supplementary data associated with this article can
References and notes
18. Keppler, D.; Sameni, M.; Moin, K.; Mikkelsen, T.; Diglio,
C. A.; Sloane, B. F. Biochem. Cell. Biol. 1996, 74, 799.
19. (a) Na¨gler, D. K.; Storer, A. C.; Portaro, F. C. V.;
`
1. Puente, X. S.; Sanchez, L. M.; Overall, C. M. Nat. Rev.
Genet. 2003, 4, 544–558.
´
Carmona, E.; Juliano, L.; Menard, R. Biochemistry 1997,
2. (a) Leung, D.; Abbenante, G.; Fairlie, D. P. J. Med.
Chem. 2000, 43, 305–341; (b) Weissleder, R.; Ntziachris-
tos, V. Nat. Med. 2003, 9, 123–128.
3. Goddard, J-P.; Reymond, J-L. Trends Biotechnol. 2004,
22, 1–15.
36, 12608; (b) Willenbrock, F.; Brocklehurst, K. Biochem.
J. 1984, 222, 805.
20. Handbook of Proteolytic Enzymes; Barret, A. J., Rawlings,
N. D., Woessner, J. F., Jr., Eds.; Academic Press: London,
1998.
4. (a) Melo, R. L.; Alves, L. C.; Del Nery, E.; Juliano, L.;
Juliano, M. A. Anal. Biochem. 2001, 293, 71–77; (b)
Szabelski, L.; Rogiewicz, M.; Wiczk, W. Anal. Biochem.
2005, 342, 20–27; (c) Burchak, O. N.; Mugherli, L.;
Chatelain, F.; Balakirev, M. Y. Bioorg. Med. Chem. 2006,
14, 2559–2568.
21. Barrett, A. J. Biochem. J. 1980, 187, 909.
22. Drouet, C.; Bouillet, L.; Csopaki, F.; Colomb, M. G.
FEBS Lett. 1999, 458, 415–418.
23. Arnon, R. In Papain, Methods in Enzymology; Perl-
mann, G., Lorand, L., Eds.; Academic Press: NewYork,
1970, pp 226.
5. Barett, A. J. Methods Enzymol. 1994, 244, 1–15.
6. Portaro, F. C. V.; Santos, A. B. F.; Cezari, M. H. S.;
Juliano, M. A.; Juliano, L.; Carmona, E. Biochem. J. 2000,
347, 123–129.
7. (a) Melo, R. L.; Barbosa Pozzo, R. C.; Alves, L. C.;
Perissutti, E.; Caliendo, G.; Santagada, V.; Juliano, L.;
Juliano, M. A. Biochim. Biophys. Acta 2001, 1547, 82–94;
(b) Birney, Y. A.; O’Connor, B. F. Protein Expr. Purif.
2001, 22, 286–298.
24. Leon, S.; Quarrell, R.; Lowe, G. Bioorg. Med. Chem. Lett.
1998, 8, 2997–3002.
25. Boes, B.; Hengel, H.; Ruppert, T.; Multhaup, G.; Koszi-
nowski, U. H.; Kloetzel, P-M. J. Exp. Med 1994, 179,
901–909.
26. Cicero, D. O.; Barbato, G.; Koch, U.; Ingallinella, P.;
Bianchi, E.; Nardi, C. M.; Steinkuhler, C.; Cortese, R.;
¨
Matassa, V.; De Francesco, R.; Pessi, A.; Bazzo, R.
J. Mol. Biol. 1999, 289, 385–396.
8. Zhu, Q.; Uttamchandani, M.; Li, D. B.; Lesaicherre, M.
L.; Yao, S. Q. Org. Lett. 2003, 5, 1257–1260.
9. (a) Leytus, S. P.; Patterson, W. L.; Mangel, W. F.
Biochem. J. 1983, 215, 253–260; (b) Boonacker, E.;
Elferink, S.; Bardai, A.; Fleischer, B.; Van Noorden, C.
J. F. J. Histochem. Cytochem. 2003, 51, 959–968.
27. Molecular Probes, The Handbook—A Guide to Fluores-
cent and Labeling Techniques, Chapter 10, 10th ed. http://
28. Tsantrizos, Y. S. Biopolymers 2004, 76, 309–323.
29. Pope, A. J.; Haupts, U. M.; Moore, K. J. Drug Discov.
Today 1999, 4, 350–362.