A. I. Vovk et al. / Bioorg. Med. Chem. Lett. 20 (2010) 483–487
487
7. (a) Chen, Y. T.; Seto, C. T. J. Med. Chem. 2002, 45, 3946; (b) Hu, X.; Stebbins, C. E.
Bioorg. Med. Chem. 2005, 13, 1101; (c) Xie, J.; Seto, C. T. Bioorg. Med. Chem. 2005,
13, 2981.
8. Phan, J.; Lee, K.; Cherry, S.; Tropea, J. E.; Burke, T. R., Jr.; Waugh, D. S.
Biochemistry 2003, 42, 13113.
9. (a) Lee, K.; Gao, Y.; Yao, Z.-J.; Phan, J.; Wu, L.; Liang, J.; Waugh, D. S.; Zhang, Z.-Y.;
Burke, T. R., Jr. Bioorg. Med. Chem. Lett. 2003, 13, 2577; (b) Lee, K.; Boovanahalli, S.
K.; Nam, K.-Y.; Kang, S.-U.; Lee, M.; Phan, J.; Wu, L.; Waugh, D. S.; Zhang, Z.-Y.; No,
K. T.; Leed, J. J.; Burke, T. R., Jr. Bioorg. Med. Chem. Lett. 2005, 15, 4037.
10. Liu, S.; Zhou, B.; Yang, H.; He, Y.; Jiang, Z.-X.; Kumar, S.; Wu, L.; Zhang, Z.-Y.
J. Am. Chem. Soc. 2008, 130, 8251.
Kononets, L. A.; Tanchuk, V. Y.; Drapailo, A. B.; Kalchenko, V. I.; Kukhar, V. P. J.
17. Gritsenko, P. G.; Lugovsky, E. V.; Kalchenko, V. I.; Komisarenko, S. V.; Koshel, T.
A.; Cherenok, S. O. Rep. Nat. Acad. Sci. Ukraine, in press.
18. Zhang, Z.-Y.; Clemens, J. C.; Schubert, H. L.; Stuckey, J. A.; Fischer, M. W. F.;
Hume, D. M.; Saper, M. A.; Dixon, J. E. Biol. Chem. 1992, 267, 23759.
19. The assay solutions contained 0.05 M BISTRIS buffer (pH 6.5), 100 mM NaCI,
0.05% Brij-35, 0.6 vol % of dimethyl sulfoxide, 0.4 mM EDTA, 1 mM DTT, p-
nitrophenyl phosphate (0.5–4.0 mM) and inhibitor. The mixtures were
preincubated for 5 min at 30 °C, and reactions were initiated by the addition
of enzyme solution. The generation of p-nitrophenol during the hydrolysis of p-
nitrophenylphosphate was measured by the increase of absorbance at 410 nm,
using a molar absorption coefficient of 18,300 Mꢀ1 cmꢀ1. The Km value was
found to be 2.5 0.3 mM. The kinetics of inhibition were studied at various
11. Sun, J.-P.; Wu, L.; Fedorov, A. A.; Almo, S. C.; Zhang, Z.-Y. J. Biol. Chem. 2003, 278,
33392.
12. (a) Frechette, R. F.; Ackerman, C.; Beers, R.; Moore, J. Bioorg. Med. Chem. Lett.
1997, 7, 2169; (b) Li, X.; Bhandari, A.; Holmes, C. P.; Szardenings, A. K. Bioorg.
Med. Chem. Lett. 2004, 14, 4301; (c) Holmes, C. P.; Li, X.; Pan, Y.; Xu, C.;
Bhandari, A.; Moody, C. M.; Miguel, J. A.; Ferla, S. W.; De Francisco, M. N.;
Frederick, B. T.; Zhou, S.; Macher, N.; Jang, L.; Irvine, J. D.; Grove, J. R. Bioorg.
Med. Chem. Lett. 2008, 18, 2719.
concentrations for each inhibitor: 1: 3
40 M, 80 M; 4: 0.2 mM, 0.4 mM, 0.6 mM; 5: 0.5
0.25 M, 0.50 M, 0.75
l
M, 6
l
M; 2: 1 M, 3
l
M, 2
l
l
l
M; 3:
M; 6:
l
l
lM, 1.0 lM, 1.5
l
l
l
M. The inhibition constants were determined
measuring initial hydrolysis rates of enzyme substrate.
20. Compounds 2, 5, and 6 were evaluated as inhibitors of human recombinant
PTPb. In the assay, 1 mM p-nitrophenyl phosphate as enzyme substrate was
preincubated for 5 min at 30 °C with varying concentrations of an inhibitor in
0.05 M BISTRIS buffer (pH 7.0) containing 100 mM NaCI, 0.05% Brij-35, 0.8 vol %
of dimethyl sulfoxide and 1 mM diethylene triamine pentaacetic acid. The
reaction was started by the addition of PTPb solution and the change in
absorbance was monitored over time at 410 nm. The value of IC50 was the
concentration of the tested compound which decreased the enzyme activity to
50% after 5 min of reaction. For competitive inhibition, Ki = 1/2 IC50 at substrate
concentration equivalent to the Km value. Under assay conditions, Km value of
the PTPb was estimated to be 1.0 0.1 mM.
21. McMartin, C.; Bohacek, R. S. J. Comput. Aided Mol. Des. 1997, 11, 333.
22. Stuckey, J. A.; Schubert, H. L.; Fauman, E. B.; Zhang, Z.-Y.; Dixon, J. E.; Saper, M.
A. Nature 1994, 370, 571.
23. Hu, X.; Stebbins, C. E. Biophys. J. 2006, 91, 948.
24. Hong, J.; Ham, S. Tetrahedron Lett. 2008, 49, 2393.
13. Iyer, S.; Younker, J. M.; Czyryca, P. G.; Hengge, A. C. Bioorg. Med. Chem. Lett.
2004, 14, 5931.
14. (a) Vovk, A.; Kalchenko, V.; Cherenok, S.; Kukhar, V.; Muzychka, O.; Lozynsky,
M. Org. Biomol. Chem. 2004, 2, 3162; (b) Cherenok, S.; Vovk, A.; Muravyova, I.;
Shivanyuk, A.; Kukhar, V.; Lipkowski, J.; Kalchenko, V. Org. Lett. 2006, 8, 549.
15. (a) Böhmer, V. Angew. Chem., Int. Ed. 1995, 34, 713–745; (b) Gutsche, C. D.
Calixarenes Revisited. In Monographs in Supramolecular Chemistry; Stoddart, J.
F., Ed.; Royal Society of Chemistry: Cambridge, 1998; (c) Calixarenes 2001;
Asfari, Z., Böhmer, V., Harrowfield, J., Vicens, J., Eds.; Kluwer Academic:
Dordrecht, 2001; (d) Calixarenes in the Nanoworld; Vicens, J., Harrowfield, J.,
Baklouti, L., Eds.; Springer: Dordrecht, 2007; (e) Rodik, R. V.; Boyko, V. I.;
Kalchenko, V. I. Curr. Med. Chem. 2009, 16, 1630.
16. (a) Almi, M.; Arduini, A.; Casnati, A.; Pochini, A.; Ungaro, R. Tetrahedron 1989,
45, 2177; (b) Kasyan, O.; Swierczynski, D.; Drapailo, A.; Suwinska, K.;
Lipkowski, J.; Kalchenko, V. Tetrahedron Lett. 2003, 44, 7167; (c) Vovk, A. I.;