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7. Thiry, A.; Dogne, J.-M.; Masereel, B.; Supuran, C. T.
Curr. Top. Med. Chem. 2007, 7, 855.
24. Fisher, S. Z.; Govindasamy, L.; Boyle, N.; Agbandje-
McKenna, M.; Silverman, D. N.; Blackburn, G. M.;
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25. Supuran, C. T.; Scozzafava, A.; Casini, A. Development
of sulfonamide carbonic anhydrase inhibitors. In Carbonic
Anhydrase–Its Inhibitors and Activators; Supuran, C. T.,
Scozzafava, A., Conway, J., Eds.; CRC Press: Boca
Raton, 2004; pp 67–147.
´
8. Thiry, A.; Dogne, J.-M.; Masereel, B.; Supuran, C. T.
Trends Pharmacol. Sci. 2006, 27, 566.
ˇ
´
9. Svastova, E.; Hulıkova, A.; Rafajova, M.; Zatovicova,
´
´
´
M.; Gibadulinova, A.; Casini, A.; Cecchi, A.; Scozzafava,
´
´
A.; Supuran, C. T.; Pastorek, J.; Pastorekova, S. FEBS
Lett. 2004, 577, 439.
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10. Pastorekova, S.; Kopacek, J.; Pastorek, J. Curr. Top. Med.
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11. Dubois, L.; Douma, K.; Supuran, C. T.; Chiu, R. K.; van
26. Boyle, N. A.; Chegwidden, W. R.; Blackburn, G. M. Org.
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Zandvoort, M. A. M. J.; Pastorekova, S.; Scozzafava, A.;
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S. D.; Liu, Y.; Hill, B.; Vullo, D.; Scozzafava, A.;
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28. 2-N,N-Dimethylamino-5-methyl-1,3,4-thiadiazole. POCl3
(0.82 ml, 8.8 mmol) was added dropwise to an ice-cooled
mixture of glacial acetic acid (0.5 ml, 8.8 mmol) and 4,4-
dimethylthiosemicarbazide (0.50 g, 4.2 mmol), under
argon. The resulting suspension was heated at 70 ꢁC for
3 h, ice-cold water (100 ml) was added, and the residue
dissolved with gentle heating and stirring. The resulting
suspension was filtered and the filtrate was adjusted to pH
8 with saturated NaHCO3. The mixture was extracted
with EtOAc (3· 50 ml) and the combined organic extracts
were washed with brine and dried over MgSO4. Evapo-
ration in vacuo gave the product as a white solid (0.49 mg,
82%); TLC (100% EtOAc): Rf = 0.2; dH (250 MHz;
CDCl3) 3.08 (6H, s, 2· NCH3), 2.53 (3H, s, CCH3); dC
(250 MHz; CDCl3) 172.1 (C3), 154.2 (C2), 41.4 (C4 and 5),
15.8 (C1); MS (EI+) m/z 143 (M+, 100%); HRMS calcd for
Wouters, B. G.; Lambin, P. Radiother. Oncol. 2007, 83,
367.
12. (a) De Simone, G.; Vitale, R. M.; Di Fiore, A.; Pedone,
C.; Scozzafava, A.; Montero, J. L.; Winum, J. Y.;
Supuran, C. T. J. Med. Chem. 2006, 49, 5544; (b) Alterio,
V.; Vitale, R. M.; Monti, S. M.; Pedone, C.; Scozzafava,
A.; Cecchi, A.; De Simone, G.; Supuran, C. T. J. Am.
Chem. Soc. 2006, 128, 8329.
13. Krungkrai, J.; Krungkrai, S. R.; Supuran, C. T. Curr. Top.
Med. Chem. 2007, 7, 909.
14. (a) Nishimori, I.; Minakuchi, T.; Morimoto, K.; Sano, S.;
Onishi, S.; Takeuchi, H.; Vullo, D.; Scozzafava, A.;
Supuran, C. T. J. Med. Chem. 2006, 49, 2117; (b)
Nishimori, I.; Minakuchi, T.; Kohsaki, T.; Onishi, S.;
Takeuchi, H.; Vullo, D.; Scozzafava, A.; Supuran, C. T.
Bioorg. Med. Chem. Lett. 2007, 17, 3585.
15. Klengel, T.; Liang, W. J.; Chaloupka, J.; Ruoff, C.;
Schroppel, K.; Naglik, J. R.; Eckert, S. E.; Morgensen, E.
G.; Haynes, K.; Tuite, M. F.; Levin, L. R.; Buck, J.;
Muhlschlegel, F. A. Curr. Biol. 2005, 15, 2021.
16. Mogensen, E. G.; Janbon, G.; Chaloupka, J.; Steegborn,
C.; Fu, M. S.; Moyrand, F.; Klengel, T.; Pearson, D. S.;
Geeves, M. A.; Buck, J.; Levin, L. R.; Muhlschlegel, F. A.
Eukaryot. Cell 2006, 5, 103.
C5H9N3S: 143.0517. Found: 143.0518; IR (KBr, cmÀ1
3414, 1554, 1420, 1337.
2-N,N-Dimethylamino-1,3,4-thiadiazole-5-methanesulfon-
)
17. Bahn, Y. S.; Muhlschlegel, F. A. Curr. Opin. Microbiol.
2006, 9, 572.
amide 5. BuLi (0.84 ml, 2.1 mmol) was added to 2-N,N-
dimethylamino-5-methyl-1,3,4-thiadiazole (0.3 g,
18. Suarez Covarrubias, A.; Larsson, A. M.; Hogbom, M.;
Lindberg, J.; Bergfors, T.; Bjorkelid, C.; Mowbray, S. L.;
Unge, T.; Jones, T. A. J. Biol. Chem. 2005, 280, 18782.
19. De Simone, G.; Di Fiore, A.; Menchise, V.; Pedone, C.;
Antel, J.; Casini, A.; Scozzafava, A.; Wurl, M.; Supuran,
C. T. Bioorg. Med. Chem. Lett. 2005, 15, 2315.
20. (a) Cecchi, A.; Hulikova, A.; Pastorek, J.; Pastorekova, S.;
Scozzafava, A.; Winum, J.-Y.; Montero, J.-L.; Supuran,
C. T. J. Med. Chem. 2005, 48, 4834; (b) Winum, J. Y.;
Temperini, C.; El Cheikh, K.; Innocenti, A.; Vullo, D.;
Ciattini, S.; Montero, J. L.; Scozzafava, A.; Supuran, C.
2.1 mmol) in anhydrous THF (10 ml), under argon, at
À78 ꢁC. Sulfur dioxide was introduced into the solution at
À78 ꢁC for 1 h, causing a precipitate and the solution to
turn purple. The mixture was concentrated in vacuo and
the residue dissolved in aq sodium acetate (20 ml, 1.1 M).
Hydroxylamine-O-sulfonic acid (1.13 g, 10 mmol) was
added and the solution stirred at rt overnight. The
solution was adjusted to pH 7 with saturated NaHCO3,
extracted with EtOAc (3· 35 ml), and the combined
organic extracts washed with brine, dried (MgSO4), and
evaporated. The resulting pale yellow solid was suspended
in warm CHCl3, collected by filtration, and washed with
CHCl3 to give the product as an off-white solid (0.12 g,
25%); TLC (100% EtOAc): Rf = 0.2 (stained yellow with
ninhydrin); mp 191–193 ꢁC (decomposition); dH
(250 MHz; DMSO-d6) 7.16 (2H, s, NH2), 4.62 (2H, s,
CH2), 3.09 (6H, s, 2· NCH3); dC (250 MHz; DMSO)-d6
172.8 (C3), 147.4 (C2), 55.3 (C1), 41.0 (C4 and 5); MS
(EI+) m/z 222 (M+, 20%); HRMS calcd for C5H10N4O2S2:
222.0245. Found: 222.0246; IR (KBr, cmÀ1) 3332, 1568,
1330, 1164, 1124.
T. J. Med. Chem. 2006, 49, 7024; (c) Gruneberg, S.;
¨
Stubbs, M. T.; Klebe, G. J. Med. Chem. 2002, 45, 3588.
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J.; Whitesides, G. M. J. Med. Chem. 1995, 38, 2286; (b)
Kim, C. Y.; Chang, J. S.; Doyon, J. B.; Baird, T. T.;
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design. In Carbonic Anhydrase—Its Inhibitors and Activa-
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22. Kim, C. Y.; Whittington, D. A.; Chang, J. S.; Liao, J.;
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23. Boriack-Sjodin, P. A.; Zeitlin, S.; Chen, H. H.; Crenshaw,
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O
O
N
N
3
5
2
1
S
N
4
NH2
S
29. Khalifah, R. G. J. Biol. Chem. 1971, 246, 2561, An
Applied Photophysics stopped-flow instrument has been
used for assaying the CA catalyzed CO2 hydration
activity. Phenol red (at a concentration of 0.2 mM) has
been used as indicator, working at the absorbance