Chem. Lett., 2010, 20, 4376; (b) J. M. Wagner, B. S. Avvaru,
A. H. Robbins, A. Scozzafava, C. T. Supuran and R. McKenna,
Bioorg. Med. Chem., 2010, 18, 4873.
10 (a) J. F. Domsic, B. S. Avvaru, C. U. Kim, S. M. Gruner,
M. Agbandje-McKenna, D. N. Silverman and R. McKenna, J. Biol.
Chem., 2008, 283, 30766; (b) D. W. Christianson and C. A. Fierke, Acc.
Chem. Res., 1996, 29, 331; (c) K. Kohler, A. Hillebrecht, J. Schulze
¨
Wischeler, A. Innocenti, A. Heine, C. T. Supuran and G. Klebe,
Angew. Chem., Int. Ed., 2007, 46, 7697; (d) A. E. Eriksson, T. A. Jones
and A. Liljas, Proteins, 1988, 4, 274.
11 J. Schulze Wischeler, A. Innocenti, D. Vullo, A. Agrawal,
S. M. Cohen, A. Heine, C. T. Supuran and G. Klebe, ChemMedChem,
2010, 5, 1609.
12 C. Temperini, A. Innocenti, A. Scozzafava and C. T. Supuran,
Bioorg. Med. Chem. Lett., 2006, 16, 4316.
13 A. Bencini, P. Failli, B. Valtancoli and D. Bani, Cardiovasc.
Hematol. Agents Med. Chem., 2010, 8, 128.
14 O. H. Szolar, Anal. Chim. Acta, 2007, 582, 191.
15 E. Amin, A. A. Saboury, H. Mansouri-Torshizi and A. A. Moosavi-
Movahedi, J. Enzyme Inhib. Med. Chem., 2010, 25, 272.
16 L. Morpurgo, A. Desideri, A. Rigo, P. Viglino and G. Rotilio,
Biochim. Biophys. Acta, 1983, 746, 168.
17 (a) A. Innocenti, A. Scozzafava and C. T. Supuran, Bioorg. Med.
Chem. Lett., 2009, 19, 1855; (b) A. Innocenti, A. Scozzafava and
C. T. Supuran, Bioorg. Med. Chem. Lett., 2010, 20, 1548;
(c) C. Temperini, A. Scozzafava and C. T. Supuran, Bioorg.
Med. Chem. Lett., 2010, 20, 474.
18 Y. Lou, P. C. McDonald, A. Oloumi, S. Chia, C. Oslund,
A. Ahmadi, A. Kyle, U. auf dem Keller, S. Leung, D. Huntsman,
B. Clarke, B. W. Sutherland, A. D. Waterhouse, M. Bally,
C. Roskelley, C. M. Overall, A. Minchinton, F. Pacchiano,
F. Carta, A. Scozzafava, N. Touisni, J. Y. Winum, C. T. Supuran
and S. Dedhar, Cancer Res., 2011, 71, 3364.
Fig. 2 View of compounds 8 (green), 9 (magenta), superposed in the
active site of hCA II. hCA II is depicted as a grey surface representation.
Amino acids labeled surfaces are colored salmon. The active-site zinc is
depicted as a blue sphere. This figure has been illustrated using PyMOL
(DeLano Scientific).
Notes and references
1 (a) C. T. Supuran, Nat. Rev. Drug Discovery, 2008, 7, 168;
(b) C. T. Supuran, Bioorg. Med. Chem. Lett., 2010, 20, 346.
2 (a) C. T. Supuran, A. Scozzafava and A. Casini, Med. Res. Rev.,
2003, 23, 146; (b) C. T. Supuran, Curr. Pharm. Des., 2010, 16, 3233.
3 (a) A. Maresca, C. Temperini, H. Vu, N. B. Pham, S. A. Poulsen,
A. Scozzafava, R. J. Quinn and C. T. Supuran, J. Am. Chem. Soc.,
2009, 131, 3057; (b) A. Maresca, C. Temperini, L. Pochet, B. Masereel,
A. Scozzafava and C. T. Supuran, J. Med. Chem., 2010, 53, 335.
4 D. Neri and C. T. Supuran, Nat. Rev. Drug Discovery, 2011, 10, 767.
5 P. Ebbesen, E. O. Pettersen, T. A. Gorr, G. Jobst, K. Williams,
J. Kienninger, R. H. Wenger, S. Pastorekova, L. Dubois, P. Lambin,
B. G. Wouters, C. T. Supuran, L. Poellinger, P. Ratcliffe,
19 V. Alterio, M. Hilvo, A. Di Fiore, C. T. Supuran, P. Pan,
S. Parkkila, A. Scaloni, J. Pastorek, S. Pastorekova, C. Pedone,
A. Scozzafava, S. M. Monti and G. De Simone, Proc. Natl. Acad.
Sci. U. S. A., 2009, 106, 16233.
20 R. G. Khalifah, J. Biol. Chem., 1971, 246, 2561. An applied
photophysics stopped-flow instrument has been used for assaying the
CA catalysed CO2 hydration activity. Phenol red (at a concentration of
0.2 mM) has been used as indicator, working at the absorbance
maximum of 557 nm, with 20 mM HEPES (pH 7.5) as buffer, and
20 mM Na2SO4 (for maintaining constant the ionic strength), following
the initial rates of the CA-catalyzed CO2 hydration reaction for a
period of 10–100 s. Stock solutions of an inhibitor (0.1 mM) were
prepared in distilled-deionized water and dilutions up to 0.01 nM were
done thereafter with distilled-deionized water. The inhibition constants
were obtained by non-linear least-squares methods6.
21 Co-crystals of the two CA II complexes were obtained using the
hanging drop vapor diffusion method. Drops of 10 mL (0.3 mM
hCA II; 0.7 mM drug; 0.1% dimethyl sulfoxide (DMSO); 0.8 M
sodium citrate; 50 mM Tris–HCl; pH 8.0) were equilibrated against
the precipitant solution (1.6 M sodium citrate; 50 mM Tris–HCl;
pH 8.0) at room temperature. Crystals were observed after 5 days.
Crystals were cryoprotected by immersion into 20% glycerol
precipitant solution and flash-cooled at 100 K. The X-ray diffrac-
tion data were collected using an R-AXIS IV++ image plate
system on a Rigaku RU-H3R Cu rotating anode operating at
50 kV and 22 mA, using Osmic Varimax HR optics. The
detector–crystal distance was set to 80 mm. The oscillation steps
were 11 with a 5 min exposure per image. Indexing, integration,
and scaling were performed using HKL2000.
A. Kanopka, A. Gorlach, M. Gasmann, A. L. Harris, P. Maxwell
¨
and A. Scozzafava, J. Enzyme Inhib. Med. Chem., 2009, 24(S1), 1.
6 (a) F. Carta, C. Temperini, A. Innocenti, A. Scozzafava, K. Kaila
and C. T. Supuran, J. Med. Chem., 2010, 53, 5511; (b) A. Maresca,
A. Scozzafava and C. T. Supuran, Bioorg. Med. Chem. Lett., 2010,
20, 7255; (c) A. Maresca and C. T. Supuran, Bioorg. Med. Chem.
Lett., 2010, 20, 4511.
7 V. Alterio, A. Di Fiore, K. D’Ambrosio, C. T. Supuran and G. De
Simone, X-Ray crystallography of CA inhibitors and its importance in
drug design, in Drug Design of Zinc-Enzyme Inhibitors: Functional,
Structural, and Disease Applications, ed. C. T. Supuran and
J. Y. Winum, Wiley, Hoboken, 2009, pp. 73–138.
8 (a) F. Pacchiano, M. Aggarwal, B. S. Avvaru, A. H. Robbins,
A. Scozzafava, R. McKenna and C. T. Supuran, Chem. Commun.,
2010, 46, 8371; (b) A. Di Fiore, S. M. Monti, M. Hilvo, S. Parkkila,
V. Romano, A. Scaloni, C. Pedone, A. Scozzafava, C. T. Supuran
and G. De Simone, Proteins, 2009, 74, 164.
9 (a) B. S. Avvaru, J. M. Wagner, A. Maresca, A. Scozzafava,
A. H. Robbins, C. T. Supuran and R. McKenna, Bioorg. Med.
c
1870 Chem. Commun., 2012, 48, 1868–1870
This journal is The Royal Society of Chemistry 2012