Novel benzenesulfonamides aryl and arylsulfone conjugates adopting tail/dual tail approaches: Synthesis, carbonic anhydrase inhibitory activity and molecular modeling studies

Article abstract of DOI:10.1016/j.ejmech.2021.113486

New series of benzenesulfonamide and benzoic acid derivatives were designed and synthesized using tail/dual tail approach to improve potency and selectivity as carbonic anhydrase inhibitors. The synthesized compounds evaluated as CAIs against isoforms hCA I, II, IV and IX with acetazolamide (AAZ) as standard inhibitor. The benzenesulfonamide derivatives 7a-d, 8a-h, 12a-c, 13a and 15a-c showed moderate to potent inhibitory activity with selectivity toward isoform hCA II, especially, compound 13a with (K_i = 7.6 nM), while the benzoic acid analogues 12d-f, 13b and 15d-f didn't show any activity except compounds 12d,f and 15e that showed weak activity. Additionally, molecular docking was performed for compounds 7a, 8a, 8e, 12a, 13a and 15a on isoform hCA I, II to illustrate the possible interaction with the active site to justify the inhibitory activity.

Full text of DOI:10.1016/j.ejmech.2021.113486

European Journal of Medicinal Chemistry 221 (2021) 113486
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Novel benzenesulfonamides aryl and arylsulfone conjugates adopting
tail/dual tail approaches: Synthesis, carbonic anhydrase inhibitory
activity and molecular modeling studies
,
, c
Assem H. Eldeeb a, Mahmoud F. Abo-Ashour a *, Andrea Angeli b, Alessandro Bonardi b
,
Deena S. Lasheen d, Eman Z. Elrazaz d, Alessio Nocentini b c, Paola Gratteri c,
,
,
**
Hatem A. Abdel-Aziz e, Claudiu T. Supuran b
a Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Egyptian Russian University, Badr City, Cairo, 11829, Egypt
b Department of NEUROFARBA, Section of Pharmaceutical and Nutraceutical Sciences, University of Florence, Polo Scientifico, Via U. Schiff 6, 50019, Sesto
Fiorentino, Firenze, Italy
c Department of NEUROFARBA, Section of Pharmaceutical and Nutraceutical Sciences, Laboratory of Molecular Modeling Cheminformatics & QSAR,
University of Florence, Polo Scientifico, Via U. Schiff 6, 50019, Sesto Fiorentino, Firenze, Italy
d Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Ain Shams University, P.O. Box 11566, Abbassia, Cairo, Egypt
e Department of Applied Organic Chemistry, National Research Center, Dokki, Cairo, 12622, Egypt
a r t i c l e i n f o
a b s t r a c t
Article history:
New series of benzenesulfonamide and benzoic acid derivatives were designed and synthesized using
tail/dual tail approach to improve potency and selectivity as carbonic anhydrase inhibitors. The syn-
thesized compounds evaluated as CAIs against isoforms hCA I, II, IV and IX with acetazolamide (AAZ) as
standard inhibitor. The benzenesulfonamide derivatives 7a-d, 8a-h, 12a-c, 13a and 15a-c showed
moderate to potent inhibitory activity with selectivity toward isoform hCA II, especially, compound 13a
with (Ki ¼ 7.6 nM), while the benzoic acid analogues 12d-f, 13b and 15d-f didn't show any activity except
compounds 12d,f and 15e that showed weak activity. Additionally, molecular docking was performed for
compounds 7a, 8a, 8e, 12a, 13a and 15a on isoform hCA I, II to illustrate the possible interaction with the
active site to justify the inhibitory activity.
Received 15 March 2021
Received in revised form
13 April 2021
Accepted 15 April 2021
Available online 27 April 2021
Keywords:
Benzenesulfonamides
Tail approaches
Dual tail approaches
Carbonic anhydrase inhibitors
Synthesis
© 2021 Elsevier Masson SAS. All rights reserved.
Molecular docking
1. Introduction
Zn2þ active site is essential for acid-base homeostasisꢀ by enzy-
matically catalyzing the conversion of carbon dioxide to carbonic
Carbonic anhydrases (CAs, EC 4.2.1.1) are pervasive metal-
loenzymes present in prokaryotes and eukaryotes [1]. 16 -CA
acid [5,6]. Human CAs are involved in a vast range of physiopath-
ological processes, including electrolytes secretion, pH and CO2
homeostasis, biosynthetic reactions, bone resorption and onco-
genesis, as a result, carbonic anhydrase suppressors are used for
management of glaucoma, edema, epilepsy, obesity and tumors
[7e14].
Sulphanilamide is one of the most classical classes acting as CAIs
[15,16]. During last few years, many approaches was adopted to
synthetize potent and selective carbonic anhydrase inhibitors as
tail/dual tail approaches[16e18]. In tail approach, sulphanilamide
was conjugated with different moieties to increase the interaction
with hydrophobic or hydrophilic parts of the active site[19,20].
Compounds IeV (Fig. 1) were designed as CAIs depending on the
last approach[21e24]. While dual tail approach depended on
a
isozymes were identified in mammals with various catalytic ac-
tivity and subcellular localization [2]. Up till now, 15 different iso-
enzymes of hCAs have been detected. Among these, 12 isoenzymes
are catalytically active with different cellular localizations (I-III,VII,
and XIII are cytosolic; IV, IX, XII, and XIV are membrane-bounded;
VA and VB are mitochondrial; and VI is secreted in milk and saliva),
while the CARPs VIII, X, and XI are catalytically inactive [3,4]. The
* Corresponding author.
** Corresponding author.
E-mail addresses: Mahmoud.Farid2020@gmail.com (M.F. Abo-Ashour), claudiu.
supuran@unifi.it (C.T. Supuran).
https://doi.org/10.1016/j.ejmech.2021.113486
0223-5234/© 2021 Elsevier Masson SAS. All rights reserved.

A.H. Eldeeb, M.F. Abo-Ashour, A. Angeli et al.
European Journal of Medicinal Chemistry 221 (2021) 113486
Fig. 1. Structures of some benzenesulfonamides as potent and selective CAIs.
interaction of the ligand with both the hydrophilic and hydro-
phobic parts of the active site seeking for selectivity and potency as
compounds VI-VII (Fig. 1)[17,18].
Based on the previous finding, this study presents some sul-
phanilamide derivatives conjugated with aryl and arylsulfone
moieties to synthesize novel potent and selective CAIs.
benzenesulfonamide 6 in AcOH to furnish the final compounds 8a-h.
In Scheme 2, sulfanilamide 9a or PABA 9b were dissolved in HCl
and diatzotized with NaNO2 to get compounds 10a,b which were
reduced by SnCl2 in HCl to furnish 4-hydrazinylbenzenesulfonamide
11a and 4-hydrazinylbenzoic acid 11b, respectively. The key in-
termediates 4a-c were refluxed with the hydrazinyl compounds
11a,b in AcOH to produce the targeted compounds 12a-f.
In Scheme 3, 1-phenyl-2-(phenylsulfonyl)ethan-1-one 4a was
reacted with diazonium salt of sulfanilamide 9a or PABA 9b in
pyridine at 0e5 ꢁC to furnish the final compounds 13a,b. on the
other hand, the key intermediates 4a-c were refluxed with
DMFDMA in dry xylene to get 3-(dimethylamino)-1-phenyl-2-
(phenylsulfonyl)prop-2-en-1-one derivatives 14a-c where, the
dimethylamino group was replaced with sulfanilamide 9a and
PABA 9b via heating in AcOH to furnish the final compounds 15a-f.
The proposed structure for compounds 7a-d, 8a-h, 12a-f, 13a,b
and 15a-f were entirely consistent with the data from their spectral
and elemental analyses. The 1H NMR spectra of compounds 7a-d,
8a-h, 12a-c, 13a and 15a-c were confirmed with appearance of D2O
Firstly, the 4-(hydrazinecarbonyl)benzenesulfonamide 6 was
conjugated with different with aryl and arylsulfone moieties to
verify their effect on selectivity, then, the phenyl group was grafted
with (F, Cl and Br) seeking enhancement the hydrophobic interac-
tion (7a-d and 8a-h). Moreover, SO2NH2 group was replaced with
COOH moiety as a biological isostere because of its different binding
manner to act as CAI to assess its effect (12d-f, 13b and 15d-f).
Additionally, the hydrazino linker replaced the hydrazido to
investigate its effect on the inhibitory activity as seen in compounds
I-IV replacement of hydrazino linker with hydrazido had a great
effect on the activity (12a-f). Furthermore, a positional replacement
carried out for phenyl hydrazine derivatives from the C]O moiety
to the active methylene to explore the ability of carbonyl group to
bind with the active site (13a,b). Finally, the reported enaminone
sulphanilamide conjugates V showed potent activity with moder-
ate selectivity on isoform IX, so, benzenesulfonyl group were added
as a hybrid to increase it (15a-f). (Fig. 2).
exchangeable peak of SO2NH2 at
7a-d, 8a-h, 12a-f, 13a,b and 15a-f revealed with presence of extra
D2O exchangeable peak for NH at 10e12 ppm. Further, compounds
12d-f, 13b and 15d-f were approved with presence peak of OH
acidic at 12.4e12.9 ppm. Also, compounds 7a-d and 12a-f were
confirmed with presence of active methylene peak at
5.2e5.5 ppm, which its absence also, confirm compounds 13a,b
and 15a-f, while compounds 8a-d showed aliphatic peak for CH3 at
2.38e2.40 ppm that confirm its proposed structure. It is note-
d 7.3e7.5 ppm, while compounds
d
d
2. Results and discussion
d
2.1. Chemistry
d
worthy that the spectra of enaminones 15a-f revealed the presence
of E/Z geometric conjugation forms as previously reported[24].
Onthe otherhand,13C NMR spectra ofcompounds8a-dconfirmed
The synthetic approaches implemented for design the last tar-
geted products (7a-d, 8a-h, 12a-f, 13a,b and 15a-f) were elucidated
in Schemes 1e3. In Scheme 1, various acetophenone derivatives 1a-
d were brominated in AcOH using Br2 to produce the phenylethyl
bromide analogues 2a-d, where the bromide moiety replaced with
sodium benzenesulfinate 3 via refluxing in absolute EtOH to furnish
the key intermediates 1-phenyl-2-(phenylsulfonyl)ethan-1-one de-
rivatives 4a-d. The carbonyl moiety of the key intermediates 4a-
d was condensed with 4-(hydrazinecarbonyl)benzenesulfonamide 6
by refluxing in AcOH to produce the final novel compounds 7a-d. On
the other hand, the appropriate analogue of acetophenone or alde-
hydes 1a-h was heated under reflux with 4-(hydrazinecarbonyl)
with presence of aliphatic peaks of eCH3 at
compounds 7a-d and 12a-f reveled with appearance of presence of
active methylene peak at 52.17e53.19 ppm. Further, compounds 7a-
d and 8a-h were approved with presence C]O peak of hydrazide
linker at 162.75-163.87 ppm, also, compounds 13a,b and 15a-f
showedC]Oof1-phenyl-2-(phenylsulfonyl)ethan-1-onederivatives
at 186.84e191.35 ppm, in addition, compounds 12d-f, 13b and 15d-f
approved with presence of C]O of COOH at 167.11e167.80 ppm.
d 14.99e15.22 ppm. While
d
d
d
d
2

A.H. Eldeeb, M.F. Abo-Ashour, A. Angeli et al.
European Journal of Medicinal Chemistry 221 (2021) 113486
Fig. 2. Design and structures of the targeted compounds 7a-d, 8a-h, 12a-f, 13a,b and 15a-f.
following order F>H>Cl>Br for compounds 8a-d,
H>F>Br>Cl for compounds 8e-h, H>F>(Cl]Br) for 7a-
d and H>Cl>Br for 12a-c and 15a-c
2.2. Biological evaluation
2.2.1. Carbonic anhydrase inhibition
ꢂ Replacement of the olefinic proton with (-CH3) or
(-SO2C6H5) moieties increased the activity for Br and Cl
derivatives, while, decreased the activity for unsubstituted
derivative.
All synthesized compounds 7a-d, 8a-h, 12a-f, 13a,b and 15a-f
were screened for their ability to suppress the physiologically
relevant hCA isoforms, hCA I, II, IV and IX were measured using the
standard inhibitor acetazolamide (AAZ) via a stopped flow CO2
hydras assay[25]. The following structure activity relationship
(SAR) can be assembled from the inhibition data mentioned in
Table 1:
ꢂ Change the hydrazido linker with hydrazino diminished the
inhibitory effect [compounds 7a-d (Ki ¼ 283.5e882.8 nM)
and 12a-c (Ki ¼ 951.2e7383 nM)].
ꢂ Positional replacement of benzensulfonamide from the
carbonyl moiety (compounds 12a Ki ¼ 951.2 nM) to the
active methylene enhanced the inhibitory activity (com-
pound 13a Ki ¼ 425.6 nM).
(i) The pervasive isoform CA I was heartily inhibited by the
synthetized compounds 7a-d, 8a-h, 12a-c, 13a and 15a-c
with inhibition constants ranging between 26.9 nM and
ꢂ While substitution of (-NHN ¼ ) linker with (-NHCH ¼ )
improved the activity (compounds 13a Ki ¼ 425.6 nM and
15a Ki ¼ 186.7 nM).
7.4
m
M, especially, compounds 8a,d,e (Ki ¼ 26.9e65.2 nM)
showed potent activity ranging from (4e9) times more active
than AAZ (Ki ¼ 250 nM), also, compounds 7a and 8c,h were
equipotent to AAZ. On the other hand, replacement of
(-SO2NH2) moiety with its isostere (COOH) abolished the
activity as seen in compounds 12e,f, 13b and 15d-f, while,
(ii) The CA II was affected efficiently by the synthetized com-
pounds, as compounds 7a-d, 8a-h, 12a, 13a and 15a showed potent
activity inhibition constants ranging between 7.6 nM to 89.2 nM,
also, compounds 12b-d,f and 15b,c,e showed moderate to weak
compound 12d showed weak activity with Ki ¼ 93.21
mM.
Noteworthy, there are many factors that had great impact on
the activity:
activity (Ki ¼ 0.42-65.44
mM), while compounds 12e, 13b and 15d,f
didn’t show any inhibitory activity up to 100
mM. Regarding to
ꢂ Grafting with diverse moieties at p-position for the phenyl
SAR:-
ring had awesome effect on the inhibitory activity with the
3

A.H. Eldeeb, M.F. Abo-Ashour, A. Angeli et al.
European Journal of Medicinal Chemistry 221 (2021) 113486
Scheme 1. Reagents and conditions: i) Br2/AcOH/stirring 2 h; ii) EtOH/reflux 2 h; iii) AcOH/reflux 4 h; iv) a- MeOH/H2SO4/reflux 10 h, b- NH2NH2$H2O (99%, 3 equ.)/reflux 4 h.
ꢂ Similar to CAI, grafting with diversified moieties at p-position
for the phenyl ring had great effect on the inhibitory activity
with the following order H>F>Cl>Br for 7a-d. While, for com-
pounds 8a-d Br moiety showed better activity than Cl and F
better than H with the following order F>H>Br>Cl. whilst,
compounds 8e-h showed different pattern as p-Cl derivative
was the most active one in this series (Ki ¼ 21.4 nM) and the un-
substituted analogue showed the least inhibitory activity and
the order was Cl>Br>F>H. on the other hand, the order for
compounds 12a-c and 15a-c was identical to CAI as following
H>Cl>Br.
(iii) The membrane bound isoform CA IV showed moderate
inhibitory activity for compounds 7a-d, 8a-h, 12a-c, 13a and
15a-c (Ki ¼ 0.09e8.33
moiety with its isostere (COOH) diminished the activity from
weak (compound 12f with Ki ¼ 18.07 M) to inactive up to
100 M (compounds 12d,e, 13b and 15d-f).
mM). While, replacement of (-SO2NH2)
m
m
According to SAR:-
ꢂ Compounds 7a-d showed the same order of activity as CAII
while, for compounds 12a-c Br moiety showed better activity
than Cl with the following order H>Br>Cl. whilst, compounds
15a-c showed different pattern as p-Br derivative was the most
ꢂ On the contrary to CAI, replacement of the olefinic proton with
(-CH3) or (-SO2C6H5) moieties decreases the inhibitory activity
for Br and Cl derivatives, while, increases activity for unsub-
stituted derivative.
active one in this series (Ki ¼ 6.7
mM) and the un-substituted
analogue showed the least inhibitory activity and the order
was Br>Cl>H.
ꢂ Similar to CAI, change the hydrazido linker with hydrazino
decreased the activity (compounds 7a-d Ki ¼ 26.9e89.2 nM and
12a-c Ki ¼ 80.5e794.2 nM).
ꢂ Interestingly, changing the olefinic proton showed different
pattern as replace the previous proton with (-CH3) decreased
the inhibitory activity. While, its replacement with (-SO2C6H5)
group improved the activity for unsubstituted and p-Br
substituted derivatives, whilst, it decreased the inhibitory ac-
tivity for p-F and p-Cl substituted analogues.
ꢂ Positional replacement of benzensulfonamide from the carbonyl
moiety (12a Ki ¼ 80.5 nM) to the active methylene (13a
Ki ¼ 7.6 nM) enhanced the inhibitory activity.
ꢂ Also, replacement of (-NHN ¼ ) linker with (-NHCH ¼ )
decreased the inhibitory activity (compounds 13a Ki ¼ 7.6 nM
and 15a Ki ¼ 40.1 nM).
ꢂ On the contrary to CAI and CAII, positional replacement of
benzensulfonamide from the carbonyl moiety to the active
4

A.H. Eldeeb, M.F. Abo-Ashour, A. Angeli et al.
European Journal of Medicinal Chemistry 221 (2021) 113486
Scheme 2. Reagents and conditions: i) AcOH/reflux 4 h; ii) NaNO2/HCl/stirring 0.5 h/0e5 ꢁC; iii) SnCl2/HCl/stirring 6 h/0e5 ꢁC.
Scheme 3. Reagents and conditions: i) a- NaNO2/HCl/stirring 0.5 h/0e5 ꢁC, b-pyridine/stirring 12 h/0e5 ꢁC; ii) DMFDMA/Xylene/reflux 15 h; iii) AcOH/reflux 4 h.
5

A.H. Eldeeb, M.F. Abo-Ashour, A. Angeli et al.
European Journal of Medicinal Chemistry 221 (2021) 113486
Table 1
Inhibitory data of human CA isoforms hCA I, hCA II, hCA IV, hCA IX with the synthesized compounds 7a-d, 8a-h,12a-f,13a,b and 15a-f determined by stopped-flow CO2 hydrase
assay, using the standard inhibitor acetazolamide (AAZ).
Comp.
KI (nM)
R
X
hCA I
hCA II
hCA IV
hCA IX
7a
7b
7c
7d
8a
–
–
H
Br
Cl
F
283.5
874.0
882.8
677.6
65.2
26.9
89.2
71.2
53.9
23.1
574.5
2289
1830
765.5
2936
768.8
289.2
818.0
1124
321.7
–
–
CH3
H
8b
8c
8d
8e
CH3
CH3
CH3
H
Br
Cl
F
629.7
291.0
48.9
26.9
42.3
45.8
21.2
30.4
7991
8335
699.3
1974
961.9
318.9
1447
869.0
H
8f
8g
8h
H
H
H
Br
Cl
F
4722
6162
260.9
951.2
7383
7373
93211
>100000
>100000
425.6
>100000
186.7
7447
7443
>100000
>100000
>100000
250.0
23.0
21.4
24.4
80.5
794.2
421.5
32633
>100000
9190
7.6
7184
2205
2092
2369
2487
908.7
2910
>100000
>100000
>100000
1605
>100000
2756
3035
3012
>100000
>100000
>100000
25.8
660.7
158.1
97.0
8167
8302
>100000
>100000
18070
2206
>100000
8381
6730
6950
>100000
>100000
>100000
74.0
12a
12b
12c
12d
12e
12f
13a
13b
15a
15b
15c
15d
15e
15f
AAZ
SO2NH2
SO2NH2
SO2NH2
COOH
COOH
COOH
SO2NH2
COOH
SO2NH2
SO2NH2
SO2NH2
COOH
COOH
COOH
–
H
Br
Cl
H
Br
Cl
–
–
>100000
40.1
H
Br
Cl
H
Br
Cl
–
9672
3016
>100000
65439
>100000
12.1
* Mean from 3 different assays, by a stopped flow technique (errors were in the range of 5e10% of the reported values).
methylene decreased the inhibitory activity as shown in com-
Table 2
pounds 12a (Ki ¼ 97.0 nM) and 13a (Ki ¼ 2.206
mM).
Selectivity ratios for the inhibition of hCA II over hCA I, IV and hCA IX for the syn-
thetized benzensulfonamide compounds 7a-d, 8a-h, 12a-c, 13a, 15a-c and AAZ.
ꢂ While, replacement of (-NHN ¼ ) linker with (-NHCH ¼ ) let to
decrease of the inhibitory activity as compounds 13a
Comp.
I/II
IV/II
IX/II
(Ki ¼ 2.206
mM) and 15a (Ki ¼ 8.381
mM).
7a
7b
7c
7d
8a
8b
8c
8d
8e
10.54
9.80
12.40
12.57
2.82
14.89
6.35
2.31
21.36
25.66
25.70
14.20
127.10
188.91
181.99
32.99
64.93
312.35
30.87
6.48
28.58
3.24
(iv) The isoform CAIX was also, moderately inhibited with the
11.49
20.85
13.93
22.74
6.96
68.25
28.59
95.87
97.76
97.09
30.89
1.14
synthetized compounds 7a-d, 8a-h, 12a-c, 13a and 15a-c
(Ki ¼ 0.28e3.03
mM). In addition, replacement of (-SO2NH2)
moiety with its isostere (COOH) rescinded the inhibitory
activity up to 100 mM (compounds 12d-f, 13b and 15d-f).
0.88
Concerning to SAR:-
8f
8g
8h
205.30
287.94
10.69
11.82
9.30
17.49
56.00
4.66
ꢂ Replacement of the olefinic proton with (-CH3) or (-SO2C6H5)
moieties improved the activity for all derivatives.
12a
12b
12c
13a
15a
15b
15c
AAZ
1.20
10.28
19.70
290.26
209.00
0.70
ꢂ Also, similar to CAI and CAII, change the hydrazido linker with
6.90
hydrazino
decreased
the
activity
(7a-d
with
211.18
68.73
0.31
1.00
2.13
Ki ¼ 289.2e1124 nM and 12a-c with Ki ¼ 908.7e2910 nM).
ꢂ In addition, positional replacement of benzensulfonamide from
the carbonyl moiety to the active methylene led to increase the
0.77
2.47
20.66
2.30
6.12
6

A.H. Eldeeb, M.F. Abo-Ashour, A. Angeli et al.
European Journal of Medicinal Chemistry 221 (2021) 113486
Table 3
index range (3.57e102.74), especially compound 7d that was the
most active and safe compounds (selectivity index ¼ 102.74),
thereby providing a high safety profile as anticancer agents.
Anti-proliferative impact of sulfonamides 7a-d, 8a-h,12a-c,13a,15a-c against breast
cancer (MCF-7) cell line, and human normal breast epithelia (MCF-10A) cell line.
Comp.
IC50
(
m
M)a
Selectivity Index
3. Molecular modeling studies
MCF-7
MCF-10A
7a
7b
7c
7d
8a
8b
8c
8d
8e
10.73 0.6
19.1 1.0
57.0 3.1
0.62 0.02
20.1 1.1
2.29 0.1
3.75 0.2
31.86 1.7
14.87 0.8
1.37 0.1
36.71 2.1
9.17 0.5
3.97 0.2
17.33 0.9
8.01 0.4
1.74 0.1
3.44 0.2
62.71 3.4
15.11 0.8
12.5 0.7
84.76 4.3
34.08 1.7
25.57 1.3
63.7 3.2
31.05 1.6
37.64 1.9
62.74 3.2
21.99 1.1
39.42 2.2
27.73 1.4
81.97 4.2
35.89 1.8
46.08 2.3
12.87 0.7
28.63 1.5
14.05 0.7
17.93 0.9
20.36 1.6
27.96 1.4
17.2 0.9
7.90
1.78
0.45
102.74
1.54
16.44
16.73
0.69
2.65
20.24
2.23
3.91
11.61
0.74
To explore in depth the binding mode of the synthesized com-
pounds within the hCA I and II active sites, docking studies were
performed on derivatives 7a, 8a, 8e, 12a, 13a and 15a.
Outcomes from docking showed that all compounds coordi-
nated the zinc ion with the deprotonated nitrogen atom of the
sulfonamide moiety. Moreover, the binding was strengthened by
two H-bonds, engaged by the sulfonamide group with the back-
bone NH and the side chain OH of T199, and further supported by
hydrophobic contacts between the phenyl ring of benzenesulfo-
namide and the lipophilic residues L198, A121 and F91.
Both the presence of two bulky tails in derivative 7a and the
small dimension of the hCA I active site forced the ligand towards
the lipophilic half of the binding pocket (Fig. 3A), leading the S]O
group of the sulfonic bridge in H-bond distance with the side chain
OH of Y204, a peculiar residue of this isoform. Moreover, the phenyl
rings is well stabilized by the lipophilic residues A135, A132 and
L131, and W5.
8f
8g
8h
12a
12b
12c
13a
15a
15b
15c
5-FU
3.57
8.07
5.21
0.32
1.85
1.38
a
IC50 values are the mean S.D. of three experiments.
The aromatic tail of the most flexible ligands 8a and 8e
accomodates in the pocket lined by hydrophobic residues A135,
V207 and L131. However, the strain determined by the presence in
8a of the methyl group pointing towards L198, is the probable cause
of 8a worse activity than 8e.
Instead, the hCA I/hCA II residue mutations make the hCA II
active site larger (F91/I91 and H67/N67) and more hydrophilic
(V62/N62) compred to hCA I, allowing the derivative 7a to orient
towards the hydrophilic half of the binding pocket. Here the com-
pounds engaged an H-bond between the sulfonic bridge S]O and
the side chain NH2 of N62 and vdW interactions between the ar-
omatic rings and the side chains of I91 and W5 (Fig. 3B). The phenyl
activity like CAI and CAII (compounds 12a Ki ¼ 2.487
Ki ¼ 1.605 M).
ꢂ While replacement of (-NHN ¼ ) linker with (-NHCH ¼ )
decreased of the inhibitory activity (13a Ki ¼ 1.605 M and 15a
Ki ¼ 2.756 M).
mM and 13a
m
m
m
Interestingly, all the synthetized benzensulfonamide com-
pounds 7a-d, 8a-h, 12a-c, 13a and 15a-c showed good to highly
selectivity to isoform hCA II over hCA I, IV and IX (Table 2). Espe-
cially, compound 13a which considered as the most potent deriv-
ative against hCA II with Ki ¼ 7.6 nM, also, it showed highly
selectivity ratio ¼ 56.0 over hCAI, 290.26 over hCA IV and 211.18
over hCA IX.
ring of derivatives 8a and 8e established
p-p staking interactions
with the phenyl ring of F131 side chain, hydrophobic contacts with
the side chain of I91, F131 and Q92, whereas the methyl portion of
8a is inevitably pushed towards V121 (Fig. 3B).
The shortening of the linker from hydrazido (7a, 8a, 8e) to
hydrazino (12a, 13a) and methylamino (15a) led the sulfonic bridge
S]O in H-bond contact with the inner side chain NH2 of Q92 or
N67 (Fig. 3C), instead of N62 (7a, Fig. 3B) and derivative 12a to
position the aromatic ring in a cleft lined by an ensemble of hy-
drophobic residues (I91, V121, F131, Q92 and P202). The elongation
from phenyl (12a) to benzoyl pendant, allow the compounds 13a
and 15a to reach the area of W5, H64 and F131 where the two ar-
2.2.2. Anti-proliferative activity
All herein synthetized sulfonamide derivatives 7a-d, 8a-h, 12a-
c, 13a and 15a-c were evaluated for their ability to induce cytotoxic
effect against breast cancer (MCF-7) cell line, under hypoxic con-
dition, adopting the colorimetric MTT assay (Table 3).
Regarding MCF-7 breast cancer cells, the tested sulfonamide
compounds 7a-d, 8a-h, 12a-c, 13a, 15a-c showed potent to weak
omatic rings form p-p staking and VdW interactions (Fig. 3C) that
activity compared with 5-FU (IC50 ¼ 12.5
8b,c,f,h, 12a,c, 13a and 15a were the most active derivatives with
IC50¼ (0.62e10.73) M. In particular, sulfonamide 7d was the most
active one with IC50 ¼ 0.62 M that is approximately 20-fold more
M that is
m
M). Compounds 7a,d,
overall stabilize the compounds.
m
4. Conclusion
m
than 5-FU, followed with compound 8f with IC50 ¼ 1.37
m
This study reports the design and synthesis of five series of
benzenesulfonamide derivatives 7a-d, 8a-h, 12a-c, 13a and 15a-c
and its isostere benzoic acid analogues 12d-f, 13b and 15d-f
adopting dual tail approach seeking for increasing the potency and
selectivity. All synthetized compounds were evaluated as CAIs
against hCA I, II, IV and IX with AAZ as stander inhibitor. The
benzenesulfonamide derivatives showed moderate to potent
inhibitory activity with selectivity toward isoform hCA II, especially,
compound 13a with Ki ¼ 7.6 nM and selectivity index ¼ 56.0 over
hCAI, 290.26 over hCA IV and 212.18 over hCA IX. On the other hand,
it's found that replacement of SO2NH2 moiety with its isostere
COOH abolished the inhibitory activity except compounds 12d,e
and 15f that showed weak activity. In addition, the anti-
approximately 9-fold more than 5-FU. Moreover, compounds 7b,
8a,e,d,g, 12b and 15c displayed moderate activity with IC50 range:
14.87e36.71
mM. While, compounds 7c and 15b had weak activity
with IC50 ¼ 57 and 62.71
mM, respectively.
On the other hand, all the synthetized sulfonamide compounds
7a-d, 8a-h, 12a-c, 13a, 15a-c were evaluated for their ability to
induce cytotoxic effect towards human normal breast epithelia
(MCF-10A) cell line under normal condition, to investigate their
safety. Thereafter, the selectivity index for each compound was
calculated (Table 3). The tested compounds showed non-significant
cytotoxic action with IC50 range (12.87e84.76
mM). Moreover,
compounds 7a,d, 8b,c,f,h, 12a,c, 13a and 15a had good selectivity
7

A.H. Eldeeb, M.F. Abo-Ashour, A. Angeli et al.
European Journal of Medicinal Chemistry 221 (2021) 113486
Fig. 3. Predicted binding mode of derivatives 7a (cyan), 8a (orange), 8e (green) within A) hCA I and B) hCA II active sites, and of C) 12a (dodger blue), 13a (spring green), 15a (purple)
within hCA II active site.
proliferative study revealed that compound 7d was the most active
and safe compounds, thereby providing a high safety profile as
anticancer agents. Finally, molecular docking was performed for
compounds 7a, 8a, 8e, 12a, 13a and 15a to illustrate the different
binding pattern within the active site of hCA I and II.
5.1.2.2. 4-(2-(1-(4-bromophenyl)-2-(phenylsulfonyl)ethylidene)hy-
drazine-1-carbonyl)benzenesulfonamide7b. White crystals, (yield
87%), m.p. 277e278 ꢁC; 1H NMR (400 MHz)
d ppm: 4.60, 5.48 (2H,
2s, CH2), 7.36 (1.7H, d, J ¼ 8.0 Hz, ArH), 7.48 (2H, s, SO2NH2), 7.58
(1.3H, s, ArH), 7.76 (2H, d, J ¼ 8.0 Hz, ArH), 7.88 (2H, d, J ¼ 8.0 Hz,
ArH), 7.96e8.01 (5H, m, ArH), 8.18 (1H, brs, ArH), 11.32 (1H, s, NH);
IR (KBr, n
cmꢀ1): 3321, 3249 (NH2, NH), 1679 (C]O) and 1337, 1304,
5. Experimental
1158, 1081 (2SO2); (ESI) m/z 534 [M ꢀ H]þ; EA Calcd.: C, 47.02; H,
3.38; N, 7.83; found C, 47.34; H, 3.58; N, 7.56.
5.1. Chemistry
5.1.2.3. 4-(2-(1-(4-chlorophenyl)-2-(phenylsulfonyl)ethylidene)hy-
5.1.1. General
drazine-1-carbonyl)benzenesulfonamide 7c. White crystals, (yield
A Stuart digital apparatus was used for determining melting
points. Shimadzu FT-IR 8400S infrared spectrophotometer was
used for recording infrared spectra. Both 1H NMR and 13C NMR
were measured using deuterated dimethylsulfoxide (DMSO‑d6) as
solvent but with variable frequency (100MHZ for 13C NMR,
400MHZ for 1H NMR) via Bruker spectrophotometer, at the Faculty
of Pharmacy, Ain-shams and Cairo Universities. Mass spectra were
recorded by TLC-MS Advion compact mass spectrometer (CMS).
Elemental analyses were implemented at the Regional Center for
Microbiology and Biotechnology, Al-Azhar University, Cairo, Egypt.
Compounds 4a-d, 6, 11a,b, 12b,c and 14a-c were prepared
depending on their reported procedure[23,26e29].
87%), m.p. 290e292 ꢁC; 1H NMR (400 MHz)
d ppm: 5.44 (2H, s, CH2),
7.40 (2H, d, J ¼ 8.0 Hz, ArH), 7.56e7.61 (4H, m, 2H, SO2NH2, 2H,
ArH), 7.68 (2H, t, J ¼ 8.0 Hz, ArH), 7.81 (1H, brs, ArH), 7.90 (3H, d,
J ¼ 8.0 Hz, ArH), 7.99 (3H, brs, ArH), 11.20 (1H, s, NH); 13C NMR
(100 MHz) d ppm: 52.93 (CH2),126.17,127.45,128.65,128.68,129.05,
129.29, 129.79, 130.93, 132.04, 134.83, 135.79, 139.27, 141.75, 142.34,
145.67, 148.96, 151.19, 154.37, 163.42 (C]O); IR (KBr,
n
cmꢀ1): 3327,
3239 (NH2, NH), 1674 (C]O) and 1341, 1303, 1158, 1091 (2SO2);
(ESI) m/z 490 [M ꢀ H]þ; EA Calcd.: C, 51.27; H, 3.69; N, 7.21; found C,
51.33; H, 3.53; N, 9.07.
5.1.2.4. 4-(2-(1-(4-flourophenyl)-2-(phenylsulfonyl)ethylidene)hy-
drazine-1-carbonyl)benzenesulfonamide 7d. White crystals, (yield
5.1.2. Synthesis of targeted compounds 7a-d
87%), m.p. 284e285 ꢁC; 1H NMR (400 MHz)
d ppm: 5.44 (2H, s, CH2),
In round conical flask, different phenyl sulfone derivative 4a-
d (1.21 mmol) was dissolved in 17 mL of acetic acid, to the previous
solution, equivalent amount of 4-(hydrazinecarbonyl)benzene-
sulfonamide 6 (0.27g, 1.23 mmol) was added. The reaction mixture
was heated under reflux for 4h, the formed precipitate was filtrated
while hot, washed with ethoxyethan and recrystallized from DMF/
MeOH mixture to furnish the targeted final compounds 7a-d.
7.18 (2H, brs, ArH), 7.56e7.60 (4H, m, 2H, SO2NH2, 2H, ArH), 7.67
(2H, t, J ¼ 8.0 Hz, ArH), 7.83 (1H, brs, ArH), 7.90 (3H, d, J ¼ 8.0 Hz,
ArH), 7.99 (3H, brs, ArH), 11.17 (1H, s, NH); 13C NMR (100 MHz)
d
ppm: 53.18 (CH2), 115.45, 115.66, 126.20, 127.72, 128.65, 129.13,
129.78, 129.93, 133.35, 133.37, 134.83, 136.69, 138.07, 139.22, 142.20,
147.33, 152.00, 152.90, 163.42 (C]O); IR (KBr,
cmꢀ1): 3317, 3220
n
(NH2, NH), 1676 (C]O) and 1335, 1305, 1162, 1091 (2SO2); EA
Calcd.: C, 53.04; H, 3.82; N, 8.84; found C, 53.63; H, 3.89; N, 8.71.
5.1.2.1. 4-(2-(1-phenyl)-2-(phenylsulfonyl)ethylidene)hydrazine-1-
carbonyl)benzenesulfonamide 7a. White crystals, (yield 84%), m.p.
284e285 ꢁC; 1H NMR (400 MHz)
d ppm: 5.43 (2H, s, CH2), 7.34e7.36
5.1.3. Synthesis of targeted compounds 8a-h, 12a-f and 15a-f
As previously mentioned for compounds 7a-d.
(2H, d, J ¼ 8.0 Hz, ArH), 7.57e7.60 (4H, m, 2H, SO2NH2, 2H, ArH),
7.67e7.70 (2H, m, ArH), 7.78 (2H, d, J ¼ 8.0 Hz, ArH), 7.91 (3H, d,
J ¼ 8.0 Hz, ArH), 8.0 (3H, brs, ArH), 11.18 (1H, s, NH); 13C NMR
5.1.3.1. 4-(2-(1-phenylethylidene)hydrazine-1-carbonyl)benzene-
(100 MHz)
d
ppm: 53.19 (CH2),126.22,127.54,128.31,128.62,128.64,
sulfonamide 8a. White powder, (yield 89%), m.p. 272e273 ꢁC; 1H
129.13, 129.77, 130.09, 131.54, 134.82, 136.81, 137.51, 139.28, 143.14,
NMR (400 MHz) d ppm: 2.40 (3H, s, CH3), 7.45 (2H, brs, ArH), 7.55
147.32, 150.22, 155.10, 158.22, 163.42 (C]O); IR (KBr,
n
cmꢀ1): 3317,
(2H, s, SO2NH2), 7.89 (2H, brs, ArH), 7.96 (3H, d, J ¼ 8.0Hz, ArH), 8.06
3237 (NH2, NH), 1677 (C]O) and 1344, 1299, 1149, 1079 (2SO2); EA
Calcd.: C, 55.13; H, 4.19; N, 9.18; found C, 55.56; H, 4.28; N, 8.97.
(2H, d, J ¼ 8.0Hz, ArH),10.96 (1H, s, NH); 13C NMR (100 MHz)
d ppm:
15.22 (CH3), 126.07, 126.98, 128.87, 129.08, 130.12, 132.25, 137.48,
8

A.H. Eldeeb, M.F. Abo-Ashour, A. Angeli et al.
European Journal of Medicinal Chemistry 221 (2021) 113486
138.43, 146.94, 149.67, 155.43, 156.76, 163.54 (C]O); IR (KBr,
n
136.63, 138.45, 144.27, 147.15, 147.81, 150.77, 153.03, 162.84 (C]O);
cmꢀ1): 3359, 3262 (NH2, NH), 1634 (C]O) and 1330, 1158 (SO2); EA
IR (KBr, n
cmꢀ1): 3311, 3211 (NH2, NH), 1679 (C]O) and 1341, 1156
Calcd.: C, 56.77; H, 4.76; N, 13.24; found C, 56.61; H, 4.85; N, 13.47.
(SO2); (ESI) m/z 336 [M ꢀ H]þ; EA Calcd.: C, 49.78; H, 3.58; N, 12.44;
found C, 49.76; H, 3.79; N, 12.62.
5.1.3.2. 4-(2-(1-(4-bromophenyl)ethylidene)hydrazine-1-carbonyl)
benzenesulfonamide 8b. White powder, (yield 89%), m.p.
5.1.3.8. 4-(2-(4-fluorobenzylidene)hydrazine-1-carbonyl)benzene-
sulfonamide 8h. White powder, (yield 89%), m.p. over 300 ꢁC; 1H
295e296 ꢁC; 1H NMR (400 MHz)
d ppm: 2.38 (3H, s, CH3), 7.53 (2H,
s, SO2NH2), 7.64 (2H, d, J ¼ 8.0 Hz, ArH), 7.82 (2H, d, J ¼ 8.0 Hz, ArH),
NMR (400 MHz)
d
ppm: 7.29 (2H,t, J ¼ 8.0 Hz, ArH), 7.54 (2H, s,
7.94 (2H, d, J ¼ 8.0 Hz, ArH), 8.04 (2H, d, J ¼ 8.0 Hz, ArH), 10.97 (1H,
SO2NH2), 7.8 (2H, t, J ¼ 8.0 Hz, ArH), 7.97 (2H, d, J ¼ 8.0 Hz, ArH),
8.08 (2H, d, J ¼ 8.0 Hz, ArH), 8.48 (1H, s, ¼CeH), 12.03 (1H, s, NH);
s, NH); 13C NMR (100 MHz)
d ppm: 14.99 (CH3), 123.71, 126.10,
129.06, 131.83, 135.35, 137.50, 140.43, 145.10, 146.86, 151.18, 153.40,
13C NMR (100 MHz)
d ppm: 116.30, 116.52, 126.29, 128.82, 129.93,
155.78, 163.87 (C]O); IR(KBr,
n
cmꢀ1): 3360, 3261 (NH2, NH), 1644
130.02, 131.07, 136.64, 144.65, 147.04, 148.16, 153.51, 155.07, 162.96
(C]O) and 1310, 1158 (SO2); (ESI) m/z 394 [M ꢀ H]þ; EA Calcd.: C,
(C]O); IR (KBr,
n
cmꢀ1): 3306, 3202 (NH2, NH), 1636 (C]O) and
45.47; H, 3.56; N, 10.60; found C, 45.27; H, 3.67; N, 10.84.
1345, 1159 (SO2); EA Calcd.: C, 52.33; H, 3.76; N, 13.08; found C,
52.21; H, 3.56; N, 12.94.
5.1.3.3. 4-(2-(1-(4-chlorophenyl)ethylidene)hydrazine-1-carbonyl)
benzenesulfonamide 8c. White powder, (yield 89%), m.p.
5.1.3.9. 4-(2-(1-phenyl-2-(phenylsulfonyl)ethylidene)hydrazineyl)
275e277 ꢁC; 1H NMR (400 MHz)
d
ppm: 2.38 (3H, s, CH3), 7.51 (2H,
benzenesulfonamide 12a. White powder, (yield 79%), m.p.
brs, ArH), 7.53 (2H, s, SO2NH2), 7.88 (2H, d, J ¼ 8.0 Hz, ArH), 7.94 (2H,
250e252 ꢁC; 1H NMR (400 MHz)
d ppm: 5.29 (2H, s, CH2), 7.15 (2H,
d, J ¼ 8.0 Hz, ArH), 8.04 (2H, d, J ¼ 8.0 Hz, ArH),10.98 (1H, s, NH); 13
C
s, SO2NH2), 7.23 (2H, d, J ¼ 8.0 Hz, ArH), 7.27e7.33 (3H, m, ArH), 7.49
(2H, t, J ¼ 8.0 Hz, ArH), 7.57 (1H, m, ArH), 7.67 (2H, d, J ¼ 8.0 Hz,
ArH), 7.74 (2H, d, J ¼ 8.0 Hz, ArH), 7.89 (2H, d, J ¼ 8.0 Hz, ArH), 10.30
NMR (100 MHz)
d ppm: 15.04 (CH3), 126.09, 128.77, 128.90, 129.10,
132.95, 134.87, 137.18, 137.30, 142.13, 146.91, 148.75, 152.58, 155.50,
163.77 (C]O); IR (KBr,
n
cmꢀ1): 3358, 3261 (NH2, NH), 1645 (C]O)
(1H, s, NH); 13C NMR (100 MHz)
d ppm: 52.41 (CH2), 112.72, 126.55,
and 1310, 1160 (SO2); EA Calcd.: C, 51.21; H, 4.01; N, 11.94; found C,
50.98; H, 4.23; N, 11.81.
127.65, 128.40, 128.52, 128.68, 129.37, 131.67, 132.53, 134.44, 134.99,
137.69, 139.71, 143.10, 145.99, 147.81; IR (KBr,
n
cmꢀ1): 3379, 3278
(NH2, NH) and 1330, 1294, 1146, 1085 (2SO2); EA Calcd.: C, 55.93; H,
4.46; N, 9.78; found C, 55.23; H, 4.69; N, 9.69.
5.1.4. 4-(2-(1-(4-fluorophenyl)ethylidene)hydrazine-1-carbonyl)
benzenesulfonamide 8d
White powder, (yield 89%), m.p. 280e282 ꢁC; 1H NMR
5.1.3.10. 4-(2-(1-phenyl-2-(phenylsulfonyl)ethylidene)hydrazineyl)
(400 MHz)
(2H, s, SO2NH2), 7.91e7.97 (4H, m, ArH), 8.05 (2H, d, J ¼ 8.0 Hz, ArH),
10.96 (1H, s, NH); 13C NMR (100 MHz)
ppm: 15.21 (CH3), 115.64,
115.85, 126.08, 126.15, 128.11, 129.07, 131.66, 134.86, 134.89, 137.40,
146.93, 155.88, 157.41, 163.63 (C]O); IR (KBr,
cmꢀ1): 3328, 3186
d
ppm: 2.39 (3H, s, CH3), 7.26 (2H, t, J ¼ 8.0 Hz, ArH), 7.54
benzoic acid 12d. White powder, (yield 82%), m.p. 269e270 ꢁC; 1H
NMR (400 MHz)
d
ppm: 5.32 (2H, s, CH2), 7.20 (2H, d, J ¼ 8.8 Hz,
d
ArH), 7.26e7.32 (3H, m, ArH), 7.47 (2H, t, J ¼ 8.0 Hz, ArH), 7.54 (1H, t,
J ¼ 7.2 Hz, ArH), 7.74 (2H, d, J ¼ 8.4 Hz, ArH), 7.81 (2H, d, J ¼ 8.4 Hz,
ArH), 7.89 (2H, d, J ¼ 6.8 Hz, ArH), 10.39 (1H, s, NH), 12.41 (1H, s,
n
(NH2, NH), 1659 (C]O) and 1335, 1160 (SO2); (ESI) m/z 334
[M ꢀ H]þ; EA Calcd.: C, 53.72; H, 4.21; N, 12.53; found C, 53.94; H,
4.34; N, 12.49.
OH); 13C NMR (100 MHz)
d ppm: 52.42 (CH2), 112.72, 121.82, 126.52,
128.36, 128.51, 128.70, 129.34, 131.36, 132.50, 134.39, 137.75, 139.75,
142.90, 146.24, 148.90, 151.67, 167.78 (C]O of COOH); IR (KBr,
n
cmꢀ1): 3380-2410 (OH), 3337 (NH), 1679 (C]O) and 1290, 1154
(SO2); (ESI) m/z 393 [M ꢀ H]þ; EA Calcd.: C, 63.95; H, 4.60; N, 7.10;
found C, 63.73; H, 3.68; N, 4.68.
5.1.3.5. 4-(2-benzylidenehydrazine-1-carbonyl)benzenesulfonamide
8e. White powder, (yield 89%), m.p. over 300 ꢁC; 1H NMR
(400 MHz) d ppm: 7.47e7.50 (3H, m, ArH), 7.55 (2H, s, SO2NH2), 7.76
(2H, d, J ¼ 8.0Hz, ArH), 7.99 (2H, d, J ¼ 8.0 Hz, ArH), 8.1 (2H, d,
5.1.3.11. 4-(2-(1-(4-bromophenyl)-2-(phenylsulfonyl)ethylidene)
J ¼ 8.0 Hz, ArH), 8.5 (1H, s, ¼CeH), 12.03 (1H, s, NH); 13C NMR
hydrazineyl)benzoic acid 12e. White powder, (yield 82%), m.p.
(100 MHz)
d
ppm: 126.28, 127.71, 128.84, 129.36, 130.81, 134.57,
273e274 ꢁC; 1H NMR (400 MHz)
d ppm: 5.27 (2H, s, CH2), 7.18 (2H,
136.77, 139.19, 147.14, 149.13, 153.90, 162.75 (C]O); IR (KBr,
n
d, J ¼ 8.0 Hz, ArH), 7.48e7.54 (4H, m, ArH), 7.57 (1H, t, J ¼ 8.0 Hz,
ArH), 7.69 (2H, d, J ¼ 8.0 Hz, ArH), 7.82 (2H, d, J ¼ 8.0 Hz, ArH), 7.87
(2H, d, J ¼ 8.0 Hz, ArH), 10.29 (1H, s, NH), 12.42 (1H, s, OH); 13C NMR
cmꢀ1): 3327, 3207 (NH2, NH), 1630 (C]O) and 1336, 1156 (SO2);
(ESI) m/z 302 [M ꢀ H]þ; EA Calcd.: C, 55.43; H, 4.32; N, 13.85; found
C, 53.64; H, 3.41; N, 4.41.
(100 MHz) d ppm: 52.21 (CH2), 112.80,121.68, 122.14,128.52, 128.73,
129.39, 131.36, 131.40, 134.39, 134.45, 137.06, 139.65, 148.64, 167.76
5.1.3.6. 4-(2-(4-bromobenzylidene)hydrazine-1-carbonyl)benzene-
(C]O of COOH); IR (KBr,
(C]O) and 1288, 1151 (SO2); EA Calcd.: C, 53.29; H, 3.62; N, 5.92;
n
cmꢀ1): 3374-2354 (OH), 3336 (NH), 1674
sulfonamide 8f. White powder, (yield 89%), m.p. over 300 ꢁC; 1H
NMR (400 MHz)
d
ppm: 7.55 (2H, s, SO2NH2), 7.66 (2H, d, J ¼ 8.0 Hz,
found C, 53.23; H, 3.80; N, 5.24.
ArH), 7.70 (2H, d, J ¼ 8.0 Hz, ArH), 7.98 (2H, d, J ¼ 8.0 Hz, ArH), 8.09
(2H, d, J ¼ 8.0 Hz, ArH), 8.45 (1H, s, ¼CeH),12.08 (1H, s, NH); 13C NMR
5.1.3.12. 4-(2-(1-(4-chlorophenyl)-2-(phenylsulfonyl)ethylidene)
(100 MHz)
d
ppm: 124.06, 126.28, 128.86, 129.55, 130.64, 132.36,
hydrazineyl)benzoic acid 12f. White powder, (yield 79%), m.p.
133.86,136.66,147.19,147.85,150.15,152.99,162.78 (C]O); IR (KBr,
n
275e276 ꢁC; 1H NMR (400 MHz)
d ppm: 5.27 (2H, s, CH2), 7.18 (2H,
cmꢀ1): 3313, 3209 (NH2, NH), 1634 (C]O) and 1339, 1156 (SO2); EA
d, J ¼ 8.0 Hz, ArH), 7.34 (2H, d, J ¼ 8.0 Hz, ArH), 7.49 (2H, t, J ¼ 8.0 Hz,
ArH), 7.57 (1H, t, J ¼ 8.0 Hz, ArH), 7.75 (2H, d, J ¼ 8.0 Hz, ArH), 7.83
(2H, d, J ¼ 8.0 Hz, ArH), 7.87 (2H, d, J ¼ 8.0 Hz, ArH), (1H, s, NH),
Calcd.: C, 43.99; H, 3.16; N, 10.99; found C, 44.16; H, 3.45; N, 10.74.
5.1.3.7. 4-(2-(4-chlorobenzylidene)hydrazine-1-carbonyl)benzene-
12.41 (1H, s, OH); 13C NMR (100 MHz)
d ppm: 52.17 (CH2), 112.81,
sulfonamide 8g. White powder, (yield 89%), m.p. over 300 ꢁC; 1H
122.12, 123.03, 128.15, 128.49, 128.64, 129.46, 131.33, 131.44, 133.04,
NMR (400 MHz)
d
ppm: 7.53 (2H, brs, ArH), 7.54 (2H, s, SO2NH2),
134.51, 136.52, 139.48, 143.47, 148.59, 150.93, 167.80 (C]O of
7.77 (2H, d, J ¼ 8.0 Hz, ArH), 7.98 (2H, d, J ¼ 8.0 Hz, ArH), 8.09 (2H, d,
COOH); IR (KBr, n
cmꢀ1): 3387-2360 (OH), 3343 (NH), 1676 (C]O)
J ¼ 8.0 Hz, ArH), 8.47 (1H, s, ¼CeH), 12.08 (1H, s, NH); 13C NMR
and 1289,1150 (SO2); (ESI) m/z 427 [M ꢀ H]þ; EA Calcd.: C, 58.81; H,
(100 MHz)
d
ppm: 126.28, 128.85, 129.34, 129.44, 133.48, 135.28,
4.00; N, 6.53; found C, 58.95; H, 3.86; N, 6.28.
9

A.H. Eldeeb, M.F. Abo-Ashour, A. Angeli et al.
European Journal of Medicinal Chemistry 221 (2021) 113486
5.1.3.13. 4-((3-oxo-3-phenyl-2-(phenylsulfonyl)prop-1-en-1-yl)
5.1.3.17. 4-((3-(4-bromophenyl)-3-oxo-2-(phenylsulfonyl)prop-1-en-
amino)benzenesulfonamide 15a. White crystals, (yield 82%), m.p.
1-yl)amino)benzoic acid 15e. White powder, (yield 79%), m.p. over
235e236 ꢁC; 1H NMR (400 MHz)
d
ppm: 7.34e7.38 (3H, m, 2H,
300 ꢁC; 1H NMR (400 MHz)
d ppm: 7.39e7.47 (3.7H, m, ArH),
SO2NH2, 1H, ArH), 7.44e7.47 (2H, m, ArH), 7.49e7.52 (3H, m, ArH),
7.58e7.65 (3H, m, ArH), 7.68e7.75 (2H, m, ArH), 7.77e7.83 (2H, m,
ArH), 8.05e8.13 (1.3H, m, ArH), 8.53 (0.7H, d, J ¼ 13.2 Hz, ArH),
7.50e7.54 (1.3H, t, J ¼ 7.6 Hz, ArH), 7.57 (2H, d, J ¼ 8.8 Hz, ArH), 7.60
(1H, t, J ¼ 8.0 Hz, ArH), 7.67 (1H, d, J ¼ 8.0Hz, ArH), 7.76 (1.3H, d,
J ¼ 8.0 Hz, ArH), 7.93e7.98 (2H, m, ArH), 8.11 (1H, d, J ¼ 8.0 Hz, ArH),
8.55 (0.7H, d, J ¼ 13.2 Hz, ArH), 10.58 (0.3, d, J ¼ 14.0 Hz, NH), 10.76
(0.7H, d, J ¼ 13.2 Hz, NH), 12.9 (1H, s, OH); 13C NMR (100 MHz)
10.56 (0.3H, d, J ¼ 14.0 Hz, NH), 10.74 (0.7H, d, J ¼ 13.2 Hz, NH); 13
C
NMR (100 MHz)
d ppm: 111.55, 113.72, 115.75, 118.21, 119.11, 127.61,
127.65, 127.78, 127.82, 128.53,128.69, 128.94, 129.06,129.33, 129.36,
129.39, 129.60, 131.60,132.40, 132.48,133.08, 133.44,133.72, 134.73,
137.91, 138.62, 138.78, 140.21, 140.47, 140.76, 142.49, 142.87, 142.95,
143.17, 144.87, 146.59, 147.85, 150.99, 189.74, 191.32 (C]O); IR (KBr,
d ppm: 111.29, 113.16, 113.31, 117.40, 117.98,118.70, 119.09, 126.19,
126.29, 127.17, 127.54, 127.61, 127.75,128.45, 129.08, 129.38,129.75,
130.70, 131.00, 131.36, 131.40, 131.46, 131.52, 131.59, 131.71, 132.07,
132.31, 133.19, 133.79, 134.59, 135.09, 137.70, 137.88, 139.63, 142.77,
143.02, 143.17, 143.70, 148.12, 150.85, 167.15, 167.24 (C]O of COOH),
n
cmꢀ1): 3318, 3229 (NH2, NH), 1631 (C]O) and 1389, 1302, 1137,
1095 (2SO2); (ESI) m/z 441 [M ꢀ H]þ; EA Calcd.: C, 57.00; H, 4.10; N,
188.82, 190.27 (C]O); IR (KBr, n
cmꢀ1): 3387-2360 (OH), 3119 (NH),
6.33; found C, 58.19; H, 4.36; N, 6.31.
1685, 1630 (2C ¼ O) and 1361, 1170 (SO2); EA Calcd.: C, 54.33; H,
3.32; N, 2.88; found C, 54.21; H, 3.17; N, 2.97.
5.1.3.14. 4-((3-(4-bromophenyl)-3-oxo-2-(phenylsulfonyl)prop-1-en-
5.1.3.18. 4-((3-(4-chlorophenyl)-3-oxo-2-(phenylsulfonyl)prop-1-en-
1-yl)amino)benzenesulfonamide 15b. White powder, (yield 79%),
1-yl)amino)benzoic acid 15f. White powder, (yield 79%), m.p. over
m.p. 280e281 ꢁC; 1H NMR (400 MHz)
d ppm: 7.35e7.40 (3H, m, 2H,
300 ꢁC; 1H NMR (400 MHz)
d ppm: 7.42e7.44 (2H, m, ArH),
SO2NH2, 1H, ArH), 7.50e7.54 (3.3H, m, ArH), 7.56e7.58 (2H, m, ArH),
7.60e7.66 (1.7H, m, ArH), 7.67e7.71 (1H, m, ArH), 7.75e7.80 (2H, m,
ArH), 7.82e7.84 (1.3H, m, ArH), 8.07 (1H, t, J ¼ 7.6 Hz, ArH), 8.54
(0.7H, d, J ¼ 13.2 Hz, ArH), 10.59 (0.3H, d, J ¼ 14.0 Hz, NH), 10.74
7.45e7.48 (2H, m, ArH), 7.49e7.54 (2.3H, m, ArH), 7.60e7.64 (1.7H,
m, ArH), 7.64e7.71 (1H, m, ArH), 7.76 (1.3H, d, J ¼ 8.0 Hz, ArH),
7.92e7.98 (2H, m, ArH), 8.11 (1H, d, J ¼ 8.0 Hz, ArH), 8.54 (0.7H, d,
J ¼ 12.4 Hz, ArH), 10.58 (0.3H, d, J ¼ 14.0 Hz, NH), 10.74 (0.7H, d,
(0.7H, d, J ¼ 13.2 Hz, NH); 13C NMR (100 MHz)
d ppm: 111.32, 113.03,
J ¼ 13.2 Hz, NH), 12.90 (1H, s, OH); 13C NMR (100 MHz)
d ppm:
113.34, 118.38, 119.24, 126.21, 126.31, 127.31, 127.55, 127.65, 127.75,
127.82, 128.45, 129.40, 129.76, 130.70, 131.43, 131.46, 131.60, 132.08,
132.32, 132.61, 132.87, 133.22, 133.81, 134.61, 135.09, 135.68, 137.68,
137.84, 140.32, 140.78, 142.34, 142.75, 142.77, 143.01, 148.30, 151.14,
111.34, 113.37, 117.94, 118.72, 127.12, 127.60, 127.62, 127.82, 128.35,
128.69, 129.13, 129.36, 130.22, 130.66, 131.12, 131.32, 131.48, 131.84,
132.25, 133.12, 133.72, 133.98, 134.50, 137.14, 137.31, 137.37, 137.62,
141.03, 142.09, 142.92, 143.18, 143.26, 143.79, 147.95, 150.81, 151.90,
188.79, 190.26 (C]O); IR (KBr, n
cmꢀ1): 3327, 3235 (NH2, NH), 1623
167.11, 167.19 (C]O of COOH), 188.56, 190.01 (C]O); IR (KBr,
n
cmꢀ1): 3313-2456 (OH), 3114 (NH), 1686, 1625 (2C ¼ O) and 1359,
1171 (SO2); (ESI) m/z 440 [M ꢀ H]þ; EA Calcd.: C, 59.80; H, 3.65; N,
3.17; found C, 59.96; H, 3.97; N, 3.38.
(C]O) and 1338, 1298, 1139, 1098 (2SO2); EA Calcd.: C, 48.38; H,
3.29; N, 5.37; found C, 48.95; H, 3.41; N, 5.48.
5.1.3.15. 4-((3-(4-chlorophenyl)-3-oxo-2-(phenylsulfonyl)prop-1-en-
5.1.5. Synthesis of targeted compounds 13a,b
1-yl)amino)benzenesulfonamide 15c. White powder, (yield 79%),
m.p. 270e271 ꢁC; 1H NMR (400 MHz)
d ppm: 7.36 (2H, s, SO2NH2),
A solution of sulphanilamide 9a or PAPA 9b (5.5 mmol) in 15 mL
of HCl was diazotized with equivalent amount of NaNO2. The pre-
vious freshly prepared solution was added gradually to solution of
1-phenyl-2-(phenylsulfonyl)ethan-1-one 4a (1.3g, 5.0 mmol) in
20 mL pyridine at (0e5) ꢁC over 33 min. The reaction mixture was
left for stirring for 12 h at 0 ꢁC then poured onto ice water, the
formed precipitate was collected and recrystallized form EtOH/
dioxane mixture to furnish the targeted final compounds 13a,b.
7.42e7.47 (2H, m, ArH), 7.48e7.51 (2H, m, ArH), 7.51e7.54 (2.3H, m,
ArH), 7.61e7.65 (2H, m, ArH), 7.66e7.70 (0.7H, m, ArH), 7.76e7.79
(1.3H, m, ArH), 7.80e7.84 (2H, m, ArH), 8.07 (1H, t, J ¼ 8.0 Hz, ArH),
8.53 (0.7H, d, J ¼ 12.8 Hz, ArH),10.59 (0.3H, d, J ¼ 14.0 Hz, NH),10.77
(0.7H, d, J ¼ 13.2 Hz, NH); 13C NMR (100 MHz)
d ppm: 111.34, 112.99,
113.36, 118.35, 119.21, 126.92, 127.55, 127.64, 127.75, 127.82, 128.45,
128.68, 129.15, 129.40, 129.76, 130.59, 131.28, 131.41, 133.22, 133.81,
137.20, 137.31, 137.48, 138.92, 140.30, 140.76, 141.38, 142.34, 142.76,
142.78, 143.02, 148.23, 148.92, 151.12, 188.65, 190.10 (C]O); IR (KBr,
5.1.4.1. 4-(2-(2-oxo-2-phenyl-1-(phenylsulfonyl)ethylidene)hydrazi-
neyl)benzenesulfonamide 13a. White powder, (yield 84%), m.p.
n
cmꢀ1): 3316, 3231 (NH2, NH), 1624 (C]O) and 1336, 1295, 1144,
240e241 ꢁC; 1H NMR (400 MHz)
d
ppm: 7.17 (2H, d, J ¼ 8.0 Hz, ArH),
1090 (2SO2); EA Calcd.: C, 52.88; H, 3.59; N, 5.87; found C, 53.13; H,
3.76; N, 5.98.
7.25 (2H, s, SO2NH2), 7.62 (2H, d, J ¼ 8.0 Hz, ArH), 7.70e7.75 (3H, m,
ArH), 7.79e7.82 (3H, m, ArH), 7.83 (2H, d, J ¼ 8.0 Hz, ArH), 7.95 (2H,
d, J ¼ 8.0 Hz, ArH), 11.58 (1H, s, NH); 13C NMR (100 MHz)
d ppm:
5.1.3.16. 4-((3-oxo-3-phenyl-2-(phenylsulfonyl)prop-1-en-1-yl)
114.46, 116.72, 127.65, 127.82, 128.68, 129.80, 129.89, 130.11, 130.29,
134.73,135.25,135.88,136.96,138.43,140.02,140.28,142.03,145.79,
amino)benzoic acid 15d. White powder, (yield 79%), m.p.
287e288 ꢁC; 1H NMR (400 MHz)
d
ppm: 7.34e7.42 (2.7H, m, ArH),
189.53 (C]O); IR (KBr, n
cmꢀ1): 3317, 3220 (NH2, NH), 1642 (C]O)
7.44e7.48 (2.3H, m, ArH), 7.50e7.54 (2H, m, ArH), 7.57e7.69 (3H, m,
ArH), 7.73 (1.3H, d, J ¼ 8.0 Hz, ArH), 7.91e7.97 (2H, m, ArH),
8.07e8.13 (1H, m, ArH), 8.53 (0.7H, d, J ¼ 13.2 Hz, ArH), 10.55 (0.3H,
d, J ¼ 14.0 Hz, NH),10.71 (0.7H, d, J ¼ 13.2 Hz, NH),12.90 (1H, s, OH);
and 1332, 1301, 1155, 1138 (2SO2); (ESI) m/z 442 [M ꢀ H]þ; EA
Calcd.: C, 54.17; H, 3.86; N, 9.48; found C, 53.97; H, 3.59; N, 9.81.
5.1.4.2. 4-(2-(2-oxo-2-phenyl-1-(phenylsulfonyl)ethylidene)hydrazi-
13C NMR (100 MHz)
d
ppm: 111.50, 113.65, 117.82, 118.56, 127.00,
neyl)benzoic acid 13b. White powder, (yield 79%), m.p.
127.53, 127.59, 127.77, 128.43, 128.54, 128.68, 129.08, 129.23, 129.34,
129.36, 129.51, 129.71, 130.99, 131.38, 131.52, 132.40, 132.49, 133.08,
133.73, 138.60, 138.78, 142.89, 143.12, 143.30, 143.82, 147.67, 150.73,
255e256 ꢁC; 1H NMR (400 MHz)
d
ppm: 7.14 (2H, d, J ¼ 8.0 Hz, ArH),
7.61 (2H, t, J ¼ 8.0 Hz, ArH), 7.69 (2H, t, J ¼ 8.0 Hz, ArH), 7.77e7.80
(2H, m, ArH), 7.83e7.88 (3H, m, ArH), 7.93e7.96 (3H, m, ArH), 11.75
167.13, 167.25 (C]O of COOH), 189.80, 191.35 (C]O); IR (KBr,
n
(1H, s, NH), 12.67 (1H, s, OH); 13C NMR (100 MHz)
d ppm: 114.29,
cmꢀ1): 3363-2434 (OH), 3128 (NH), 1684, 1626 (2C ¼ O) and 1357,
1174 (SO2); EA Calcd.: C, 64.85; H, 4.21; N, 3.44; found C, 65.37; H,
4.37; N, 3.51.
116.42, 117.84, 121.71, 125.28, 127.01, 127.28, 127.64, 128.64, 128.68,
128.71, 129.80, 129.88, 130.10, 130.29, 131.33, 131.47, 132.19, 133.30,
133.98, 134.70, 135.11, 135.30, 135.83, 136.97, 138.57, 140.09, 140.19,
10

A.H. Eldeeb, M.F. Abo-Ashour, A. Angeli et al.
European Journal of Medicinal Chemistry 221 (2021) 113486
properties of novel urea and sulfamide derivatives incorporating dopami-
nergic 2-aminotetralin scaffolds, Bioorg. Med. Chem. 24 (2016) 2318e2329.
[15] C.T. Supuran, How many carbonic anhydrase inhibition mechanisms exist?
J. Enzym. Inhib. Med. Chem. 31 (2016) 345e360.
[16] V. Alterio, A. Di Fiore, K. D'Ambrosio, C.T. Supuran, G. De Simone, Multiple
binding modes of inhibitors to carbonic anhydrases: how to design specific
drugs targeting 15 different isoforms? Chem. Rev. 112 (2012) 4421e4468.
[17] R.P. Tanpure, B. Ren, T.S. Peat, L.F. Bornaghi, D. Vullo, C.T. Supuran, S.-
A. Poulsen, Carbonic anhydrase inhibitors with dual-tail moieties to match the
hydrophobic and hydrophilic halves of the carbonic anhydrase active site,
J. Med. Chem. 58 (2015) 1494e1501.
[18] M. Fares, W.M. Eldehna, S. Bua, C. Lanzi, L. Lucarini, E. Masini, T.S. Peat,
H.A. Abdel-Aziz, A. Nocentini, P.A. Keller, C.T. Supuran, Discovery of potent
dual-tailed benzenesulfonamide inhibitors of human carbonic anhydrases
implicated in glaucoma and in vivo profiling of their intraocular pressure-
lowering action, J. Med. Chem. 63 (2020) 3317e3326.
140.38, 141.44, 142.85, 143.88, 144.51, 145.61, 146.80, 148.73, 167.19,
167.37 (C]O of COOH), 186.84, 189.59 (C]O); IR (KBr,
3324-2523 (OH), 3211 (NH), 1684, 1646 (2C ¼ O) and 1312, 1166
(SO2); (ESI) m/z 407 [M ꢀ H]þ; EA Calcd.: C, 61.76; H, 3.95; N, 6.86;
found C, 61.68; H, 3.96; N, 6.95.
n
cmꢀ1):
5.2. Biological evaluation
The used procedure performed as reported for carbonic anhy-
drase inhibitory activity [30e32] and anti-proliferative study
[33e35] mentioned in the Supplementary Materials.
[19] W.M. Eldehna, M.F. Abo-Ashour, A. Nocentini, R.S. El-Haggar, S. Bua,
A. Bonardi, S.T. Al-Rashood, G.S. Hassan, P. Gratteri, H.A. Abdel-Aziz,
C.T. Supuran, Enhancement of the tail hydrophobic interactions within the
carbonic anhydrase IX active site via structural extension: design and syn-
thesis of novel N-substituted isatins-SLC-0111 hybrids as carbonic anhydrase
inhibitors and antitumor agents, Eur. J. Med. Chem. 162 (2019) 147e160.
[20] A. Scozzafava, L. Menabuoni, F. Mincione, F. Briganti, G. Mincione,
C.T. Supuran, Carbonic anhydrase inhibitors. Synthesis of water-soluble, top-
ically effective, intraocular pressure-lowering aromatic/heterocyclic sulfon-
amides containing cationic or anionic Moieties: is the tail more important
than the ring? J. Med. Chem. 42 (1999) 2641e2650.
5.3. Molecular modelling
The procedures applied in the docking experiments for com-
pounds 7a, 8a, 8e, 12a,13a and 15a within CA I (PDB 3K34) and CA II
(PDB 3K34) were provided in the supplementary materials
[36e42].
Declaration of competing interest
[21] M.F. Abo-Ashour, W.M. Eldehna, A. Nocentini, A. Bonardi, S. Bua, H.S. Ibrahim,
The authors declare that they have no known competing
financial interests or personal relationships that could have
appeared to influence the work reported in this paper.
ꢂ
M.M. Elaasser, V. Krystof, R. Jorda, P. Gratteri, S.M. Abou-Seri, C.T. Supuran, 3-
Hydrazinoisatin-based benzenesulfonamides as novel carbonic anhydrase
inhibitors endowed with anticancer activity: synthesis, in vitro biological
evaluation and in silico insights, Eur. J. Med. Chem. 184 (2019) 111768.
[22] H.A. Allam, S.H. Fahim, M.F. Abo-Ashour, A. Nocentini, M.E. Elbakry,
M.A. Abdelrahman, W.M. Eldehna, H.S. Ibrahim, C.T. Supuran, Application of
hydrazino and hydrazido linkers to connect benzenesulfonamides with hy-
drophilic/phobic tails for targeting the middle region of human carbonic
anhydrases active site: selective inhibitors of hCA IX, Eur. J. Med. Chem. 179
(2019) 547e556.
[23] M.F. Abo-Ashour, W.M. Eldehna, A. Nocentini, H.S. Ibrahim, S. Bua, S.M. Abou-
Seri, C.T. Supuran, Novel hydrazido benzenesulfonamides-isatin conjugates:
synthesis, carbonic anhydrase inhibitory activity and molecular modeling
studies, Eur. J. Med. Chem. 157 (2018) 28e36.
Appendix A. Supplementary data
Supplementary data to this article can be found online at
https://doi.org/10.1016/j.ejmech.2021.113486.
References
[24] W.M. Eldehna, M.F. Abo-Ashour, E. Berrino, D. Vullo, H.A. Ghabbour, S.T. Al-
Rashood, G.S. Hassan, H.M. Alkahtani, A.A. Almehizia, A. Alharbi, H.A. Abdel-
Aziz, C.T. Supuran, SLC-0111 enaminone analogs, 3/4-(3-aryl-3-oxopropenyl)
aminobenzenesulfonamides, as novel selective subnanomolar inhibitors of
the tumor-associated carbonic anhydrase isoform IX, Bioorg. Chem. 83 (2019)
549e558.
[1] S. Del Prete, D. Vullo, V. De Luca, Z. AlOthman, S.M. Osman, C.T. Supuran,
C. Capasso, Biochemical characterization of recombinant
b-carbonic anhy-
drase (PgiCAb) identified in the genome of the oral pathogenic bacterium
Porphyromonas gingivalis, J. Enzym. Inhib. Med. Chem. 30 (2015) 366e370.
[2] C.T. Supuran, Carbonic anhydrases-an overview, Curr. Pharmaceut. Des. 14
(2008) 603e614.
[25] R.G. Khalifah, The carbon dioxide hydration activity of carbonic anhydrase I.
Stop-flow kinetic studies on the native human isoenzymes B and C, J. Biol.
Chem. 246 (1971) 2561e2573.
[26] H.S. Ibrahim, H.A. Allam, W.R. Mahmoud, A. Bonardi, A. Nocentini, P. Gratteri,
E.S. Ibrahim, H.A. Abdel-Aziz, C.T. Supuran, Dual-tail arylsulfone-based ben-
zenesulfonamides differently match the hydrophobic and hydrophilic halves
of human carbonic anhydrases active sites: selective inhibitors for the tumor-
associated hCA IX isoform, Eur. J. Med. Chem. 152 (2018) 1e9.
[27] Soliman, Preparation and antidiabetic activity of some sulfonylurea de-
rivatives of 3, 5-disubstituted pyrazoles, J. Med. Chem. 22 (1979) 321e325.
[28] H.A. Abdel-Aziz, K.A. Al-Rashood, K.E.H. ElTahir, G.M. Suddek, Synthesis of N-
benzenesulfonamide-1H-pyrazoles bearing arylsulfonyl moiety: novel cele-
coxib analogs as potent anti-inflammatory agents, Eur. J. Med. Chem. 80
(2014) 416e422.
[3] C.T. Supuran, A. Scozzafava, m. chemistry, Carbonic anhydrases as targets for
medicinal chemistry, Bioorg. Med. Chem. 15 (2007) 4336e4350.
[4] C.T. Supuran, Carbonic anhydrases as drug targets-an overview, Curr. Top.
Med. Chem. 7 (2007) 825e833.
ꢀ
[5] J. Chiche, M.C. Brahimi-Horn, J. Pouyssegur, Tumour hypoxia induces
a
metabolic shift causing acidosis: a common feature in cancer, J. Cell Mol. Med.
14 (2010) 771e794.
[6] H.-P. Schneider, M.D. Alt, M. Klier, A. Spiess, F.T. Andes, A. Waheed, W.S. Sly,
H.M. Becker, J.W. Deitmer, GPI-anchored carbonic anhydrase IV displays both
intra-and extracellular activity in cRNA-injected oocytes and in mouse neu-
rons, Proc. Natl. Acad. Sci. Unit. States Am. 110 (2013) 1494e1499.
[7] C.T. Supuran, Carbonic anhydrases: novel therapeutic applications for in-
hibitors and activators, Nat. Rev. Drug Discov. 7 (2008) 168e181.
[8] F. Carta, C.T. Supuran, Diuretics with carbonic anhydrase inhibitory action: a
patent and literature review (2005e2013), Expert Opin. Ther. Pat. 23 (2013)
681e691.
[29] M. Takahashi, T. Mamiya, H. Hasegawa, T. Nagai, H. Wakita, Preparation of 1,5-
disubstituted 4-sulfonylpyrazoles,
substituted 4-sulfonylisoxazoles
b
-cyano-
from
b
-sulfonylenamines and 5-
b
-keto- -sulfonylenamines,
b
[9] C.T. Supuran, A. Scozzafava, A. Casini, Carbonic anhydrase inhibitors, Med. Res.
Rev. 23 (2003) 146e189.
J. Heterocycl. Chem. 23 (1986) 1363e1366.
[30] M.F. Abo-Ashour, W.M. Eldehna, A. Nocentini, H.S. Ibrahim, S. Bua, H.A. Abdel-
Aziz, S.M. Abou-Seri, C.T. Supuran, Novel synthesized SLC-0111 thiazole and
thiadiazole analogues: determination of their carbonic anhydrase inhibitory
activity and molecular modeling studies, Bioorg. Chem. 87 (2019) 794e802.
[31] W.M. Eldehna, M.F. Abo-Ashour, A. Nocentini, P. Gratteri, I.H. Eissa, M. Fares,
O.E. Ismael, H.A. Ghabbour, M.M. Elaasser, H.A. Abdel-Aziz, Novel 4/3-((4-oxo-
5-(2-oxoindolin-3-ylidene) thiazolidin-2-ylidene) amino) benzenesulfona-
mides: synthesis, carbonic anhydrase inhibitory activity, anticancer activity
and molecular modelling studies, Eur. J. Med. Chem. 139 (2017) 250e262.
[32] A.A.M. Alkhaldi, M.M. Al-Sanea, A. Nocentini, W.M. Eldehna, Z.M. Elsayed,
A. Bonardi, M.F. Abo-Ashour, A.K. El-Damasy, M.S. Abdel-Maksoud, T. Al-
Warhi, P. Gratteri, H.A. Abdel-Aziz, C.T. Supuran, R. El-Haggar, 3-
[10] A. Thiry, J.-M. Dogne, B. Masereel, C.T. Supuran, Targeting tumor-associated
carbonic anhydrase IX in cancer therapy, Trends Pharmacol. Sci. 27 (2006)
566e573.
_
[11] M. Boztas¸ , Y. Çetinkaya, M. Topal, I. Gülçin, A. Menzek, E. S¸ ahin, M. Tanc,
C.T. Supuran, Synthesis and carbonic anhydrase isoenzymes I, II, IX, and XII
inhibitory effects of dimethoxybromophenol derivatives incorporating
cyclopropane moieties, J. Med. Chem. 58 (2015) 640e650.
_
ꢁ
ꢁ
[12] A. Akıncıoglu, E. Kocaman, H. Akıncıoglu, R.E. Salmas, S. Durdagi, I. Gülçin,
€
C.T. Supuran, S. Goksu, The synthesis of novel sulfamides derived from
b-
benzylphenethylamines as acetylcholinesterase, butyrylcholinesterase and
carbonic anhydrase enzymes inhibitors, Bioorg. Chem. 74 (2017) 238e250.
_
[13] I. Gülçin, B. Trofimov, R. Kaya, P. Taslimi, L. Sobenina, E. Schmidt, O. Petrova,
Methylthiazolo[3,2-a]benzimidazole-benzenesulfonamide
conjugates
as
S. Malysheva, N. Gusarova, V. Farzaliyev, A. Sujayev, S. Alwasel, C.T. Supuran,
Synthesis of nitrogen, phosphorus, selenium and sulfur-containing hetero-
cyclic compounds e determination of their carbonic anhydrase, acetylcho-
novel carbonic anhydrase inhibitors endowed with anticancer activity:
design, synthesis, biological and molecular modeling studies, Eur. J. Med.
Chem. 207 (2020) 112745.
linesterase, butyrylcholinesterase and
Bioorg. Chem. 103 (2020) 104171.
[14] B. Ozgeris¸ , S. Goksu, L. Polat Kose, I. Gülçin, R.E. Salmas, S. Durdagi, F. Tümer,
C.T. Supuran, Acetylcholinesterase and carbonic anhydrase inhibitory
a-glycosidase inhibition properties,
[33] M. Shaldam, W.M. Eldehna, A. Nocentini, Z.M. Elsayed, T.M. Ibrahim, R. Salem,
R.A. El-Domany, C. Capasso, H.A. Abdel-Aziz, C.T. Supuran, Development of
novel benzofuran-based SLC-0111 analogs as selective cancer-associated
€
_
€
€
11

A.H. Eldeeb, M.F. Abo-Ashour, A. Angeli et al.
European Journal of Medicinal Chemistry 221 (2021) 113486
sciences, Nucleic Acids Res. 49 (2021) D437eD451.
carbonic anhydrase isoform IX inhibitors, Eur. J. Med. Chem. 216 (2021)
113283.
[38] C.A. Behnke, I. Le Trong, J.W. Godden, et al., Atomic resolution studies of
[34] A. Petreni, A. Bonardi, C. Lomelino, S.M. Osman, Z.A. Alothman, W.M. Eldehna,
carbonic anhydrase II, Acta Crystallogr. D Biol. Crystallogr. 66 (2010) 616e627.
[39] Schrodinger Suite Release 2019-1, vol. 5, Schrodinger, LLC, New York, NY,
2019, p. 5 (a) Prime.(b) Maestro v.11.9;.
€
€
R. El-Haggar, R. McKenna, A. Nocentini, C.T. Supuran, Inclusion of
a
5-
fluorouracil moiety in nitrogenous bases derivatives as human carbonic
anhydrase IX and XII inhibitors produced a targeted action against MDA-MB-
231 and T47D breast cancer cells, Eur. J. Med. Chem. 190 (2020) 112112.
[35] M.M. Elbadawi, W.M. Eldehna, A. Nocentini, M.F. Abo-Ashour, E.B. Elkaeed,
M.A. Abdelgawad, K.S. Alharbi, H.A. Abdel-Aziz, C.T. Supuran, P. Gratteri,
M.M. Al-Sanea, Identification of N-phenyl-2-(phenylsulfonyl)acetamides/
propanamides as new SLC-0111 analogues: synthesis and evaluation of the
carbonic anhydrase inhibitory activities, Eur. J. Med. Chem. 218 (2021)
113360.
[36] D.K. Srivastava, K.M. Jude, A.L. Banerjee, M. Haldar, S. Manokaran, J. Kooren,
S. Mallik, D.W. Christianson, Structural analysis of charge discrimination in the
binding of inhibitors to human carbonic anhydrases I and II, J. Am. Chem. Soc.
129 (2007) 5528e5537.
(c) Epik, v.4.7;.
(d) Impact, v.8.2;.
(e) Macromodel v.12.3..
(f) Glide, v.8.2;.
(g) Desmond, v.5.7;.
(h) Jaguar, v. 10.3..
[40] A. Pustenko, A. Nocentini, P. Gratteri, et al., The antibiotic furagin and its
derivatives are isoform-selective human carbonic anhydrase inhibitors,
J. Enzym. Inhib. Med. Chem. 35 (2020) 1011e1020.
[41] A. Bonardi, A. Nocentini, R. Cadoni, et al., Benzoxaboroles: new potent in-
hibitors of the carbonic anhydrases of the pathogenic bacterium Vibrio
cholerae, ACS Med. Chem. Lett. 11 (2020) 2277e2284.
[42] A. Nocentini, A. Bonardi, P. Gratteri, et al., Steroids interfere with human
carbonic anhydrase activity by using alternative binding mechanisms,
J. Enzym. Inhib. Med. Chem. 33 (2018) 1453e1459.
[37] RCSB Protein Data Bank: powerful new tools for exploring 3D structures of
biological macromolecules for basic and applied research and education in
fundamental biology, biomedicine, biotechnology, bioengineering and energy
12