Synthesis of rhodamine B-benzenesulfonamide conjugates and their inhibitory activity against human α- And bacterial/fungal β-carbonic anhydrases

Article abstract of DOI:10.1016/j.bmcl.2011.07.045

A series of fluorescent sulfonamide carbonic anhydrase (CA, EC 4.2.1.1) inhibitors were obtained by attaching rhodamine B moieties to the scaffold of benzenesulfonamides. The new compounds have been investigated for the inhibition of 12 human α-CA isoform

Full text of DOI:10.1016/j.bmcl.2011.07.045

Bioorganic & Medicinal Chemistry Letters 21 (2011) 5210–5213
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
Synthesis of rhodamine B–benzenesulfonamide conjugates and their inhibitory
activity against human
a- and bacterial/fungal b-carbonic anhydrases
Marouan Rami a, Alessio Innocenti b, Jean-Louis Montero a, Andrea Scozzafava b,
b,
⇑
Jean-Yves Winum a, , Claudiu T. Supuran
⇑
a Institut des Biomolécules Max Mousseron (IBMM), UMR 5247 CNRS-UM1-UM2, Bâtiment de Recherche Max Mousseron, Ecole Nationale Supérieure de Chimie de Montpellier,
8 rue de l’Ecole Normale, 34296 Montpellier Cedex, France
b Università degli Studi di Firenze, Laboratorio di Chimica Bioinorganica, Rm. 188, Via della Lastruccia 3, I-50019 Sesto Fiorentino (Firenze), Italy
a r t i c l e i n f o
a b s t r a c t
Article history:
A series of fluorescent sulfonamide carbonic anhydrase (CA, EC 4.2.1.1) inhibitors were obtained by
attaching rhodamine B moieties to the scaffold of benzenesulfonamides. The new compounds have been
investigated for the inhibition of 12 human a-CA isoforms (hCA I–hCA XIV), three bacterial and one fun-
gal b-class enzymes from the pathogens Mycobacterium tuberculosis and Candida albicans. All types of
inhibitory activities have been detected, with several compounds showing low nanomolar inhibition
against the transmembrane isoforms hCA IX, XII (cancer-associated) and XIV. The b-CAs were inhibited
in the micromolar range by these compounds which may have applications for the imaging of hypoxic
tumors or bacteria due to their fluorescent moieties.
Received 21 June 2011
Revised 11 July 2011
Accepted 12 July 2011
Available online 21 July 2011
Keywords:
Carbonic anhydrase
Sulfonamide
Ó 2011 Elsevier Ltd. All rights reserved.
Rhodamine B
Fluorescence
Tumor imaging
Bacterial/fungal carbonic anhydrase
Rhodamine dyes are extensively used in biotechnological appli-
cations such as fluorescence microscopy, flow cytometry, fluores-
cence correlation spectroscopy,1,2 as well as for staining micro-
organisms (such a Mycobacterium tuberculosis),3 due to their high
fluorescence quantum yields and rather simple chemical structure.
Together with fluorescein,4 these are the two dyes mostly used for
attaching fluorescent tags to biomolecules, such as for example en-
zyme inhibitors, PET tracers, agents to visualize mitochondrial
function, hypoxic tumors, etc.5,6
sulfonamide B inhibits in vivo the growth of primary tumors and
metastases in a highly aggressive breast cancer cell line, in animals
harboring such tumors.8
S
HN
O
N
H
n
SO2NH2
Ac
N N
We have explored earlier4 the use of fluorescein moieties for
preparing inhibitors of the zinc enzyme carbonic anhydrase (CA,
EC 4.2.1.1).7 Sulfonamides bearing such moieties, of types A and
B, were showing enhanced affinity for the tumor-associated iso-
forms CA IX and XII compared to the cytosolic ones CA I and II,
and were used in the proof-of-concept studies which demon-
strated the involvement of CA IX in tumor acidification processes.4b
Furthermore, in vivo, in animal models of hypoxic tumors,
compounds A and B were observed to accumulate only in the
tumor6c,6d making this type of derivatives (and the entire class of
the CA IX-selective inhibitors) interesting imaging candidates of
this type of tumors. More recently, it has been also shown that
COOH
N
H
SO2NH2
S
AZA
HO
O
A: n = 1
B: n = 2
All these data clearly show the usefulness of fluorescently
labeled CA inhibitors (CAIs) for both in vitro and in vivo studies
with this class of pharmacological agents which have various
applications as diuretics, antiglaucoma, antiobesity, antiepileptic
and antitumor agents.7,8 Furthermore, as CAs belonging to various
⇑
Corresponding authors. Tel.: +33 4 67 14 72 34; fax: +33 4 67 14 43 44 (J.Y.W.);
tel.: +39 055 457 3005; fax: +39 055 457 3385 (C.T.S.).
classes (
a-, b- and/or c-CA family) are present in many pathogenic
E-mail addresses:
jean-yves.winum@univ-montp2.fr (J.-Y. Winum), claudiu.
nematodes, bacteria and fungi,7,9,10 fluorescently labeled CAIs may
supuran@unifi.it (C.T. Supuran).
0960-894X/$ - see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmcl.2011.07.045

M. Rami et al. / Bioorg. Med. Chem. Lett. 21 (2011) 5210–5213
5211
Table 1
Inhibition data of human
have interesting applications as research tools for better under-
standing the role these enzymes play in the life cycle of these
pathogens or for eventually developing alternative pharmacologic
agents to the clinically used compounds which led in many cases
to extensive drug resistance problems.9,10
a
- and bacterial/fungal b-CAs with sulfonamides 3–6 and
acetazolamide AAZ as standard, by a stopped-flow CO2 hydrase assay.12 hCA = human
CA isoform, mtCA = Mycobacterium tuberculosis CA, caNce103 = Candida albicans
enzyme
Isoform/inhibitor
KIa (nM)
As only fluorescein-tagged CAIs have been reported up until
now,4 in this Letter we report the synthesis and inhibition studies
of CAIs of the sulfonamide type labeled with rhodamine B moieties.
For this scope, the carboxylic acid moiety of rhodamine B 1 has
been transformed to the corresponding acyl chloride by reaction
with phosphorus oxychloride, and then coupled with amino-ben-
zenesulfonamide derivatives of type 2, bearing a free amino, meth-
ylamino or ethylamino moiety. The sulfonamide–rhodamine B
conjugates 3–6 obtained in this way (Scheme 1) were thoroughly
characterized and their structures were confirmed.11
3
4
5
6
AZA
hCA I
hCA II
512
279
32,000
7200
413
310
980
60.2
18.5
9.1
280
7.7
4410
414
342
311
714
980
692
955
18,900
7500
996
250
12
20,000
74
63
54
11
2.5
25
5.7
17
41
21.5
34,500
6740
347
321
543
12.1
24.0
10.5
236
8.4
4360
443
hCA III
hCA IV
hCA VA
hCA VB
hCA VI
hCA VII
hCA IX
hCA XII
hCA XIII
hCA XIV
mtCA 1
mtCA 2
mtCA 3
caNce103
21,300
7450
1040
513
1035
34.9
10.2
8.6
515
4.9
4480
411
469
427
814
91.7
19.4
25.3
463
14.6
3700
409
The new compounds reported here, of types 3–6 and acetazol-
amide AZA as standard drug have been investigated for the inhibi-
tion of 12 human a
-CA isoforms (hCA I–hCA XIV),7 three bacterial
481
9
104
132
and one fungal b-class enzymes from the pathogens Mycobacterium
tuberculosis (mtCA 1, 2 and 3) and Candida albicans (caNce 103).9,10
Data of Table 1 show the following structure–activity relationship
(SAR) for the inhibition of these enzymes with the investigated
sulfonamides:
413
4095
422
4730
4990
5320
a
From three different determinations. Errors were in the range of 5–10% of the
reported value.
compound was the metanilamide conjugate 4, whereas the
best inhibitor was the aminoethylbenzenesulfonamide
derivative 6 (and acetazolamide AZA).
(i) The human isoforms hCA I, VA, VB, VI and XIII were inhibited
by the new sulfonamides 3–6 moderately, with inhibition
constants in the low micromolar–submicromolar range.
Thus, for hCA I, the KIs were in the range of 311–714 nM,
for hCA VA in the range of 347–1040 nM; for hCA VB in
the range of 310–513 nM; in the range of 543–1035 nM
against hCA VI; and for hCA XIII in the range of 236–
515 nM, respectively, (Table 1). It may be observed that
most of the time the metanilamide derivative 4 was the
most active in the series, followed by the sulfanilamide
one 3. The longer molecules incorporating aminomethyl
and aminoethyl linkers (5 and 6) were generally the least
active inhibitors in this small series of investigated com-
pounds. Irrespective of the fact that some of these isoforms
are cytosolic (hCA I and XIII), mitochondrial (hCA VA and
VB) or secreted (hCA VI), their behavior against this class
of inhibitors is rather similar.
(iv) The membrane-associated hCA IV was also rather weakly
inhibited by sulfonamides 3–6, with KIs in the range of
6.74–7.50 lM (Table 1). AZA on the other hand is a potent
inhibitor of this isoform (KI of 74 nM).
(v) The brain-associated, cytosolic isoform hCA VII was effec-
tively inhibited by sulfonamides 3–6 incorporating rhoda-
mine B moieties, with KIs in the range of 12.1–91.7 nM.
Again the best inhibitor was the metanilamide conjugate 4,
but followed by the aminomethyl derivative 5, whereas
the weakest one was the aminoethyl derivative 6.
(vi) The most interesting inhibition profile with derivatives 3–6
has been observed against the transmembrane isoforms
hCA IX, XII and XIV (two of them, hCA IX and XII are associ-
ated to tumors, whereas hCA XIV is not).7 Indeed, against
hCA IX these compounds showed KIs in the range of 10.2–
24.0 nM, with the metanilamide conjugate 4 being the least
effective inhibitor and the aminomethyl one 5 the best.
However, the SAR is rather flat as all these compounds are
highly effective as CA IX inhibitors (similar to AZA). Against
hCA XII the efficacy was again excellent, with KIs in the
range of 8.6–25.3 nM and a similar SAR. hCA XIV was inhib-
ited with KIs in the range of 4.9–14.6 nM, being the most
sensitive isoform to this type of CA inhibitor.
(ii) The physiologically dominant isoform hCA II was potently
inhibited by the metanilamide rhodamide conjugate 4 (KI
of 21.5 nM) and weakly inhibited by the remaining com-
pounds 3, 5 and 6, which showed KIs in the range of 279–
980 nM. The SAR is thus rather similar with what discussed
above for the isoforms I, VA, VB and XIII.
(iii) The cytosolic low activity isoforms hCA III was poorly inhib-
ited by all these sulfonamides (and AZA), with KIs in the
range of 18.9–34.5
lM. However, in this case, the least active
R2
R1
O
COOH
POCl3, NEt3
R2
N
n
N
O
N+
N
O
N
R1
Cl-
1
n
0
0
1
2
R1
R2
H2N
n
3
4
5
6
-SO2NH2 -H
2
-H
-SO2NH2
-SO2NH2 -H
-SO2NH2 -H
Scheme 1. Preparation of rhodamine-substituted sulfonamides 3–6.

5212
M. Rami et al. / Bioorg. Med. Chem. Lett. 21 (2011) 5210–5213
Schlicker, C.; Hall, R. A.; Vullo, D.; Middelhaufe, S.; Gertz, M.; Supuran, C. T.;
Muhlschlegel, F. A.; Steegborn, C. J. Mol. Biol. 2009, 385, 1207.
(vii) The mycobacterial enzyme mtCA 1 was weakly inhibited by
sulfonamides 3–6, with KIs in the range of 3.70–4.48 M,
l
10. (a) Syrjänen, L.; Tolvanen, M.; Hilvo, M.; Olatubosun, A.; Innocenti, A.;
Scozzafava, A.; Leppiniemi, J.; Niederhauser, B.; Hytönen, V. P.; Gorr, T. A.;
Parkkila, S.; Supuran, C. T. BMC Biochem. 2010, 11, 28; (b) Minakuchi, T.;
Nishimori, I.; Vullo, D.; Scozzafava, A.; Supuran, C. T. J. Med. Chem. 2009, 52,
2226; (c) Nishimori, I.; Minakuchi, T.; Vullo, D.; Scozzafava, A.; Innocenti, A.;
Supuran, C. T. J. Med. Chem. 2009, 52, 3116; (d) Burghout, P.; Vullo, D.;
Scozzafava, A.; Hermans, P. W. M.; Supuran, C. T. Bioorg. Med. Chem. 2011, 19,
243.
11. The rhodamine B–benzenesulfonamides conjugates 3–6 were synthesized as
follows: Rhodamine B (1 mmol) 1 was dissolved in phosphorus oxychloride
POCl3 (27 mmol). The mixture was refluxed overnight. After cooling at room
temperature, the solvent was evaporated under reduced pressure. The
obtained rhodamine B acid chloride was dissolved in acetonitrile. (10 mL).
Benzenesulfonamide (2.5 mmol) derivatives 2 and triethylamine (3.6 mmol)
were added to the solution and stirred at room temperature overnight under
N2. The mixture was then concentrated under reduced pressure, water was
added to the residue, and the aqueous phase was extracted with methylene
chloride (2 Â 40 mL). The organic layer was washed twice with water, dried
over anhydrous Na2SO4, and concentrated under vacuum. The residue was
subjected to column chromatography (silica, CH2Cl2/MeOH, 99:1).
whereas the remaining two b-CAs from this pathogen were
at least one order of magnitude more sensitive to be inhib-
ited by these compounds. Indeed, against mtCA 2 the inhibi-
tion constants were in the range of 409–443 nM, and against
mtCA 3 in the range of 342–469 nM (Table 1). Thus, the new
sulfonamides reported here are less effective than AZA as
mtCA inhibitors.
(viii) The fungal enzyme from C. albicans caNce 103 was also
weakly inhibited by the compounds investigated here,
which showed KIs in the range of 4.09–5.32 lM.
In conclusion, we report the synthesis of a series of fluorescent
CA inhibitors, which were obtained by attaching rhodamine B moi-
eties to the scaffold of benzenesulfonamides. The new compounds
have been investigated for the inhibition of 12 human
a-CA iso-
Rhodamine B-sulfanilamide conjugate 3: Yield 29%, mp: 135–137 °C, 1H NMR
(400 MHz, DMSO-d6) d 1.05 (t, 12H, J = 6.8 Hz), 3.29 (m, 8H), 6.35–6.59 (m, 6H),
7.06–7.07 (m, 1H), 7.13 (d, 2H, J = 8.2 Hz) 7.26 (s, 2H, SO2NH2), 7.55–7.59 (m,
2H), 7.6 (d, 2H, J = 8.2 Hz), 7.9 (m, 1H). 13C NMR (101 MHz, DMSO-d6) d 12.23,
43.59, 66.40, 123.0, 123.64, 125.43, 124.87, 125.96, 126.08, 128.14, 128.55,
128.67, 133.77, 139.96, 140.89, 141.41, 151.42, 151.95, 167.05, MS (ESI+/ESIÀ)
m/z: 597.44 [M+H]+, 619.21 [M+Na]+, 595.37 [MÀH]À.
forms (hCA I–hCA XIV), three bacterial and one fungal b-class en-
zymes from the pathogens M. tuberculosis and C. albicans. All
types of inhibitory activities have been detected, with several com-
pounds showing low nanomolar inhibition against the transmem-
brane isoforms hCA IX, XII (cancer-associated) and XIV. The b-CAs
were inhibited in the micromolar range by these compounds
which may have applications for the imaging of hypoxic tumors
or bacteria due to their fluorescent moieties.
Rhodamine B-3-aminobenzenesulfonamide conjugate 4: Yield 32%, mp: 137–
139 °C, 1H NMR (400 MHz, DMSO-d6) d 1.05 (t, 12H, J = 6.8 Hz), 3.31 (m, 8H),
6.36–6.38 (m, 2H), 6.57 (s, 2H), 6.87–6.89 (m, 2H), 7.07 (d, 1H, J = 6.7 Hz), 7.38
(s, 2H), 7.36 (m, 2H), 7.55–7.65 (m, 4H), 7.91 (d, 1H, J = 6.7 Hz). 13C NMR
(101 MHz, DMSO-d6) d: 12.06, 45.23, 66.40, 115.81, 122.96, 123.19, 123.49,
128.83, 129.07, 129.23, 133.72, 133.92, 136.67, 137.17, 144.48, 151.95, 166.94,
MS (ESI+/ESIÀ) m/z: 597.44 [M+H]+, 595.44 [MÀH]À.
Acknowledgment
Rhodamine B-4-aminomethylbenzenesulfonamide conjugate 5: Yield 16%, mp
>205 °C, 1H NMR (400 MHz, DMSO-d6) d 1.06 (t, 12H, J = 6.8 Hz), 3.29 (q, 8H,
J = 6.9 Hz), 4.15 (s, 2H), 6.18 (m, 4H), 6.23 (s, 2H), 7.0 (d, 2H, J = 8.2 Hz), 7.09 (s,
1H), 7.15 (s, 2H), 7.44 (d, 2H, J = 8.2 Hz), 7.53–7.54 (m, 2H) 7.81–7.84 (m, 1H).
13C NMR (101 MHz, DMSO-d6) d 12.27, 42.85, 43.62, 64.37, 104.49, 96.92,108.
122.34, 123.73, 124.90, 128.06, 128.37, 128.52, 130.55, 132.81, 141.42, 142.11,
148.29, 152.72, 166.76, MS (ESI+/ESIÀ) m/z: 611.44 [M+H]+, 633.46 [M+Na]+,
609.39 [MÀH]À, 645.39 [M+Cl]À.
This research was financed in part by a 7 FP EU project (Metoxia).
References and notes
1. (a) Karstens, T.; Kobs, K. J. Phys. Chem. 1980, 84, 1871; (b) Setiawan, D.;
Kazaryan, A.; Martoprawiro, M. A.; Filatov, M. Phys. Chem. Chem. Phys. 2010, 12,
11238; (c) Kubin, R. J. Luminesc. 1983, 27, 455.
2. (a) Würth, C.; Grabolle, M.; Pauli, J.; Spieles, M.; Resch-Genger, U. Anal. Chem.
2011, 83, 3431; (b) Hossen, M. N.; Kajimoto, K.; Akita, H.; Hyodo, M.; Ishitsuka,
T.; Harashima, H. J. Controlled Release 2010, 147, 261.
3. (a) Stancu, M. M.; Grifoll, M. J. Gen. Appl. Microbiol. 2011, 57, 1; (b) Hoff, D. R.;
Ryan, G. J.; Driver, E. R.; Ssemakulu, C. C.; De Groote, M. A.; Basaraba, R. J.;
Lenaerts, A. J. PLoS One 2011, 6, e17550.
Rhodamine B-4-aminoethylbenzenesulfonamide conjugate 6: Yield 61%, mp: 117–
119 °C, 1H NMR (400 MHz, DMSO-d6) d 1.08 (t, 12H, J = 6.8 Hz), 2.48 (t, 2H,
J = 8.2 Hz), 3.33 (m, 10H), 6.27–6.42 (m, 6H), 7.03 (d, 2H, J = 8.2 Hz) 7.07–7.09
(m, 1H), 7.26 (s, 2H), 7.52–7.55 (m, 2H), 7.62 (d, 2H, J = 8.2 Hz), 7.78–7.81
(m, 1H). 13C NMR (101 MHz, DMSO-d6) d: 12.27, 33.69, 43.61, 45.49, 64.10,
97.10, 108.16,122.22, 104.94, 123.63, 125.64, 128.34, 128.57, 130.84, 132.60,
132.96, 142.01, 142.93, 148.34, 152.81, 166.33, MS (ESI+/ESIÀ) m/z: 625.44
[M+H]+, 647.48 [M+Na]+, 623.39 [MÀH]À.
4. (a) Cecchi, A.; Supuran, C. T. Curr. Pharm. Des. 2008, 14, 699; (b) Švastová, E.;
´
ˇ
Hulıková, A.; Rafajová, M.; Zatovicová, M.; Gibadulinová, A.; Casini, A.; Cecchi,
A.; Scozzafava, A.; Supuran, C.; Pastorek, J. FEBS Lett. 2004, 577, 439; (c) Cecchi,
A.; Hulikova, A.; Pastorek, J.; Pastoreková, S.; Scozzafava, A.; Winum, J.-Y.;
Montero, J.-L.; Supuran, C. T. J. Med. Chem. 2005, 48, 4834; (d) 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.
12. Khalifah, R. G. 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 10–20 mM Hepes (pH
7.5, for the a-CAs) or TRIS (pH 8.3, for the b-CAs) as buffers, and 20 mM Na2SO4
or 20 mM NaClO4 (for maintaining constant the ionic strength), following the
initial rates of the CA-catalyzed CO2 hydration reaction for a period of 10–
100 s. The CO2 concentrations ranged from 1.7 to 17 mM for the determination
of the kinetic parameters and inhibition constants. For each inhibitor at least
six traces of the initial 5–10% of the reaction have been used for determining
the initial velocity. The uncatalyzed rates were determined in the same manner
and subtracted from the total observed rates. Stock solutions of inhibitor
(1 mM) were prepared in distilled–deionized water and dilutions up to 0.1 nM
were done thereafter with distilled-deionized water. Inhibitor and enzyme
solutions were preincubated together for 15 min at room temperature prior to
assay, in order to allow for the formation of the E–I complex. The inhibition
constants were obtained by non-linear least-squares methods using PRISM 3,
and the Cheng-Prosogg equation, as reported earlier,8,10 and represent the
mean from at least three different determinations. All CA isoforms were
recombinant ones obtained in house as reported earlier.13–16
5. a Zhou, Y.; Kim, Y. S.; Yan, X.; Jacobson, O.; Chen, X.; Liu, S. Mol. Pharm. in press.;
(b) Koide, Y.; Urano, Y.; Hanaoka, K.; Terai, T.; Nagano, T. ACS Chem. Biol. 2011,
6, 600.
6. (a) Best, Q. A.; Xu, R.; McCarroll, M. E.; Wang, L.; Dyer, D. J. Org. Lett. 2010, 12,
3219; (b) Gottumukkala, V.; Heinrich, T. K.; Baker, A.; Dunning, P.; Fahey, F. H.;
Treves, S. T.; Packard, A. B. Nucl. Med. Biol. 2010, 37, 365; (c) Dubois, L.; Douma,
K.; Supuran, C. T.; Chiu, R. K.; van Zandvoort, M. A. M. J.; Pastoreková, S.;
Scozzafava, A.; Wouters, B. G.; Lambin, P. Radiother. Oncol. 2007, 83, 367; (d)
Dubois, L.; Lieuwes, N. G.; Maresca, A.; Thiry, A.; Supuran, C. T.; Scozzafava, A.;
Wouters, B. G.; Lambin, P. Radiother. Oncol. 2009, 92, 423.
7. (a) Supuran, C. T. Nat. Rev. Drug Disc. 2008, 7, 168; (b) Supuran, C. T. Bioorg. Med.
Chem. Lett. 2010, 20, 3467.
8. (a) Lou, Y.; McDonald, P. C.; Oloumi, A.; Chia, S. K.; Ostlund, C.; Ahmadi, A.; Kyle,
A.; Auf dem Keller, U.; Leung, S.; Huntsman, D. G.; Clarke, B.; Sutherland, B. W.;
Waterhouse, D.; Bally, M. B.; Roskelley, C. D.; Overall, C. M.; Minchinton, A.;
Pacchiano, F.; Carta, F.; Scozzafava, A.; Touisni, N.; Winum, J. Y.; Supuran, C. T.;
Dedhar, S. Cancer Res. 2011, 71, 3364; (b) Pacchiano, F.; Carta, F.; McDonald, P.
C.; Lou, Y.; Vullo, D.; Scozzafava, A.; Dedhar, S.; Supuran, C. T. J. Med. Chem.
2011, 54, 1896.
9. (a) Hall, R. A.; Mühlschlegel, F. A. Fungal and Nematode Carbonic Anhydrases:
Their Inhibition in Drug Design. In Drug Design of Zinc-Enzyme Inhibitors:
Functional, Structural, and Disease Applications; Supuran, C. T., Winum, J. Y., Eds.;
John Wiley & Sons: Hoboken, 2009; pp 301–322; (b) Ohndorf, U. M.; Schlicker,
C.; Steegborn, C. Crystallographic Studies on Carbonic Anhydrases from Fungal
Pathogens for Structure-Assisted Drug Development. In Drug Design of Zinc-
Enzyme Inhibitors: Functional, Structural, and Disease Applications; Supuran, C. T.,
13. (a) Lehtonen, J. M.; Shen, B.; Vihinen, M.; Casini, A.; Scozzafava, A.; Supuran, C.
T.; Parkkila, A.-K.; Saarnio, J.; Kivelä, A. J.; Waheed, A.; Sly, W. S.; Parkkila, S. J.
Biol. Chem. 2004, 279, 2719; (b) Pastorekova, S.; Parkkila, S.; Pastorek, J.;
Supuran, C. T. J. Enzyme Inhib. Med. Chem. 2004, 19, 199; (c) Abbate, F.; Winum,
J. Y.; Potter, B. V. L.; Casini, A.; Montero, J. L.; Scozzafava, A.; Supuran, C. T.
Bioorg. Med. Chem. Lett. 2004, 14, 231; (d) Clare, B. W.; Supuran, C. T. J. Pharm.
Sci. 1994, 83, 768.
14. (a) Winum, J. Y.; Temperini, C.; El Cheikh, K.; Innocenti, A.; Vullo, D.; Ciattini, S.;
Montero, J. L.; Scozzafava, A.; Supuran, C. T. J. Med. Chem. 2006, 49, 7024; (b)
Casini, A.; Scozzafava, A.; Mincione, F.; Menabuoni, L.; Ilies, M. A.; Supuran, C. T.
J. Med. Chem. 2000, 43, 4884; (c) Casey, J. R.; Morgan, P. E.; Vullo, D.;
Scozzafava, A.; Mastrolorenzo, A.; Supuran, C. T. J. Med. Chem. 2004, 47,
2337.
Winum, J. Y., Eds.; John Wiley
& Sons: Hoboken, 2009; pp 323–334; (c)

M. Rami et al. / Bioorg. Med. Chem. Lett. 21 (2011) 5210–5213
5213
15. (a) Nishimori, I.; Vullo, D.; Innocenti, A.; Scozzafava, A.; Mastrolorenzo, A.;
Supuran, C. T. J. Med. Chem. 2005, 48, 7860; (b) Nishimori, I.; Minakuchi, T.;
Onishi, S.; Vullo, D.; Cecchi, A.; Scozzafava, A.; Supuran, C. T. Bioorg. Med. Chem.
2007, 15, 7229; (c) Supuran, C. T. Curr. Pharm. Des. 2008, 14, 641; (d) Supuran,
C. T.; Vullo, D.; Manole, G.; Casini, A.; Scozzafava, A. Curr. Med. Chem.—
Cardiovasc. Hematol. Agents 2004, 2, 49.
16. (a) Nishimori, I.; Vullo, D.; Innocenti, A.; Scozzafava, A.; Mastrolorenzo, A.;
Supuran, C. T. Bioorg. Med. Chem. Lett. 2005, 15, 3828; (b) Vullo, D.; Voipio, J.;
Innocenti, A.; Rivera, C.; Ranki, H.; Scozzafava, A.; Kaila, K.; Supuran, C. T.
Bioorg. Med. Chem. Lett. 2005, 15, 971; (c) Clare, B. W.; Supuran, C. T. Eur. J. Med.
Chem. 1999, 34, 463; (d) Supuran, C. T.; Popescu, A.; Ilisiu, M.; Costandache, A.;
Banciu, M. D. Eur. J. Med. Chem. 1996, 31, 439.