Synthesis and biological evaluation of aminomethyl and alkoxymethyl derivatives as carbonic anhydrase, acetylcholinesterase and butyrylcholinesterase inhibitors

Article abstract of DOI:10.1080/14756366.2017.1368019

Compounds containing nitrogen and sulfur atoms can be widely used in various fields such as industry, medicine, biotechnology and chemical technology. Therefore, the reactions of aminomethylation and alkoxymethylation of mercaptobenzothiazole, mercaptobenzoxazole and 2-aminothiazole were developed. Additionally, the alkoxymethyl derivatives of mercaptobenzoxazole and 2-aminothiazole were synthesized by a reaction with hemiformals, which are prepared by the reaction of alcohols and formaldehyde. In this study, the inhibitory effects of these molecules were investigated against acetylcholinesterase (AChE), butyrylcholinesterase (BChE) enzymes and carbonic anhydrase I, and II isoenzymes (hCA I and II). Both hCA isoenzymes were significantly inhibited by the recently synthesized molecules, with K_i values in the range of 58–157 nM for hCA I, and 81–215 nM for hCA II. Additionally, the K_i parameters of these molecules for BChE and AChE were calculated in the ranges 23–88 and 18–78 nM, respectively.

Full text of DOI:10.1080/14756366.2017.1368019

Journal of Enzyme Inhibition and Medicinal Chemistry
ISSN: 1475-6366 (Print) 1475-6374 (Online) Journal homepage: http://www.tandfonline.com/loi/ienz20
Synthesis and biological evaluation of
aminomethyl and alkoxymethyl derivatives as
carbonic anhydrase, acetylcholinesterase and
butyrylcholinesterase inhibitors
İlhami Gulçin , Malahat Abbasova, Parham Taslimi, Zübeyir Huyut, Leyla
Safarova, Afsun Sujayev, Vagif Farzaliyev, Şükrü Beydemir, Saleh H. Alwasel
& Claudiu T. Supuran
To cite this article: İlhami Gulçin , Malahat Abbasova, Parham Taslimi, Zübeyir Huyut,
Leyla Safarova, Afsun Sujayev, Vagif Farzaliyev, Şükrü Beydemir, Saleh H. Alwasel
& Claudiu T. Supuran (2017) Synthesis and biological evaluation of aminomethyl and
alkoxymethyl derivatives as carbonic anhydrase, acetylcholinesterase and butyrylcholinesterase
inhibitors, Journal of Enzyme Inhibition and Medicinal Chemistry, 32:1, 1174-1182, DOI:
10.1080/14756366.2017.1368019
To link to this article: http://dx.doi.org/10.1080/14756366.2017.1368019
© 2017 The Author(s). Published by Informa
UK Limited, trading as Taylor & Francis
Group.
Published online: 11 Sep 2017.
Submit your article to this journal
View related articles
View Crossmark data
Full Terms & Conditions of access and use can be found at
http://www.tandfonline.com/action/journalInformation?journalCode=ienz20
Download by: [Australian Catholic University]
Date: 12 September 2017, At: 02:11

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY, 2017
VOL. 32, NO. 1, 1174–1182
https://doi.org/10.1080/14756366.2017.1368019
RESEARCH PAPER
Synthesis and biological evaluation of aminomethyl and alkoxymethyl derivatives
as carbonic anhydrase, acetylcholinesterase and butyrylcholinesterase inhibitors
a,b
c
a
d
c
c
_
€
, Malahat Abbasova , Parham Taslimi , Zubeyir Huyut , Leyla Safarova , Afsun Sujayev ,
€ €
Ilhami Gulc¸in
c
e
b
f,g
Vagif Farzaliyev , S¸ukru Beydemir , Saleh H. Alwasel and Claudiu T. Supuran
b
aDepartment of Chemistry, Faculty of Sciences, Ataturk University, Erzurum, Turkey; Department of Zoology, College of Science, King Saud
c
University, Riyadh, Saudi Arabia; Laboratory of Theoretical Bases of Synthesis and Action Mechanism of Additives, Institute of Chemistry of
d
€ €
€
Additives, Azerbaijan National Academy of Sciences, Baku, Azerbaijan; Department of Biochemistry, Faculty of Medical, Yuzuncu Yıl University,
e
f
Van, Turkey; Department of Biochemistry, Faculty of Pharmacy, Anadolu University, Eskis¸ehir, Turkey; Dipartimento di Chimica Ugo Schiff,
Universita degli Studi di Firenze, Florence, Italy; Neurofarba Department, Section of Pharmaceutical and Nutriceutical Sciences, Universita degli
g
Studi di Firenze, Florence, Italy
ABSTRACT
ARTICLE HISTORY
Received 21 March 2017
Revised 13 August 2017
Accepted 13 August 2017
Compounds containing nitrogen and sulfur atoms can be widely used in various fields such as industry,
medicine, biotechnology and chemical technology. Therefore, the reactions of aminomethylation and
alkoxymethylation of mercaptobenzothiazole, mercaptobenzoxazole and 2-aminothiazole were developed.
Additionally, the alkoxymethyl derivatives of mercaptobenzoxazole and 2-aminothiazole were synthesized
by a reaction with hemiformals, which are prepared by the reaction of alcohols and formaldehyde. In this
study, the inhibitory effects of these molecules were investigated against acetylcholinesterase (AChE),
butyrylcholinesterase (BChE) enzymes and carbonic anhydrase I, and II isoenzymes (hCA I and II). Both hCA
isoenzymes were significantly inhibited by the recently synthesized molecules, with Ki values in the range
of 58–157 nM for hCA I, and 81–215 nM for hCA II. Additionally, the Ki parameters of these molecules for
BChE and AChE were calculated in the ranges 23–88 and 18–78 nM, respectively.
KEYWORDS
Acetylcholinesterase;
butyrylcholinesterase;
carbonic anhydrase;
mercaptobenzothiazole;
mercaptobenzoxazole
Introduction
The production of novel CA inhibitors (CAIs) is a growing prior-
ity for pharmaceutical research and discovery. In addition to the
defined role of CAIs as antiglaucoma drugs and diuretics, their
potential as anti-obesity, anti-convulsant, anti-infective and anti-
cancer has been recently described.17,18 hCA II inhibitors has been
widely studied from structural and design points-of-view and in
dynamics simulations.19–21 In addition, it is the most widespread
physiologically relevant CA isoenzyme.
Alzheimer’s disease (AD) is the most prevalent cause of demen-
tia in elderly people.22–24 Recoveries in cognitive capabilities in AD
patients were obtained by disrupting or blocking the acetylcholin-
esterase (AChE) activity with inhibitor compounds.25–27 Alkaloid
compounds are some of the strongest acetylcholinesterase inhibi-
tors (AChEIs); therefore searches for novel alkaloids with inhibitory
compounds have been conducted.28–30 The AChE enzyme by
prompting hydrolyzes of the neurotransmitter acetylcholine (ACh),
concluding an impulse transmissions at the cholinergic synapses
in neurons.31,32 As can be seen in Figure 1, the active site of
AChE’s consists of two parts: (i) the anionic part that accommo-
dates the positively charged section of acetylcholine and (ii) the
catalytic part where the ester bond is hydrolysed.33,34 AChE is the
target of many drugs and neurotoxins that bind particularly to its
active site.35,36 Inhibition of AChE is used for the treatment of
senile dementia, AD, myasthenia gravis, ataxia and Parkinson’s dis-
ease.37–39 AChE can also serve as a probe for biosensors that are
capable of binding to and potentially discovering new AChE
inhibitor compounds; these compounds have applications as
Chemists are interested in derivatives of mercaptobenzothiazole
and mercaptobenzoxazole because a number of biologically and
physiologically active compounds with bactericidal, fungicidal,
tuberculostatic, anti-inflammatory, parasympatholytics and anes-
thetic properties have been synthesized based on them.1
The carbonic anhydrases (CAs, E.C.4.2.1.1) are a superfamily of
metalloenzymes that catalyze a crucial and simple biochemical
reaction, the reversible hydration of carbon dioxide (CO2) and
water (H2O) to bicarbonate (HCOꢀ3 ) and protons (Hþ).2–5 This reac-
tion, in the absence of CA cannot proceed with a perceptible rate
under physiological positions.6–8
CA
CO2 þ H2O () H2CO3 () HCOꢀ3 þ Hþ
CAs are widely distributed in all kingdoms of life and are cate-
gorized in seven distinct classes: a-, b-, c-, d-, f-, g- and h-CAs.
Each CA family demonstrates proper specific characteristics in the
primary amino acid sequence.9,10 a-CAs are found in mammals.
a-CAs, which have sixteen isoenzymes are expressed predomin-
antly in vertebrates and are the only class observed in humans.
They are catalytically active and differ in their subcellular localiza-
tion, distribution in organs and tissues, kinetic properties, expres-
sion levels, and inhibitor binding affinities.11–13 Additionally, CAs
play important roles in a multitude of physiological activities in
eukaryotes, such as CO2 transport, respiration, photosynthesis and
electrolyte secretion.14–16
CONTACT Ilhami Gulcin
igulcin@atauni.edu.tr
Department of Chemistry, Faculty of Sciences, Ataturk University, Erzurum, Turkey; Claudiu T. Supuran
claudiu.supuran@unifi.it
Dipartimento di Chimica Ugo Schiff, Universita degli Studi di Firenze, Florence, Italy
ß 2017 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distri-
bution, and reproduction in any medium, provided the original work is properly cited.

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
1175
Figure 1. The hydrolyze reaction of acetylcholine in the presence of acetylcholinesterase enzyme (AChE).
possible neurotoxins, such as nerve factors, pesticides and thera- mercaptobenzoxazole (or 2-aminothiazole) dissolved in ethanol
peutic drugs.40,41 X-ray structures have indicated that the although
the butyrylcholinesterase (BChE) and AChE structures are similar,
multiple structural discrepancies in the active-site gorges and the
active sites have been observed.42,43 BChE has of toxicological and
pharmacological importance because it scavenges ChEIs, including
potent organophosphorus nerve factors, before they bind synap-
ses and hydrolyzes ester-containing drugs44. BChE is also import-
ant for drug metabolism such as cocaine.45 Both BChE and AChE,
which have molecular roles beyond normal neurons and differenti-
ated kinetics recorded in the brain, accumulate within tangles and
amyloid plaques.46
was added to hemiformal at the temperature of 10 ꢁC. The result-
ing reaction water was separated by azeotropic distillation with
benzene. The crystalline substances were obtained after distilling
off the solvent (ethanol, benzene) and recrystallization.
Biological studies
Purification of carbonic anhydrase I and II isoforms and inhibition
studies
To observe of inhibition effects of novel aminomethyl and alkoxy-
methyl derivatives (1–17) on CA I, and II isoforms, which purified
from fresh human erythrocyte using an affinity chromatography
procedure.54,55 CA activity was determined using the previously
described spectrophotometric procedure of Verpoorte et al.56 as
explained previously.21,57,58 In this procedure, changes in activity
were obtained during 3 min at 22 ꢁC. p-Nitrophenylacetate (PNA)
compound was used as a substrate, and it was converted by both
isoforms to p-nitrophenolate ions.59,60 The quantity of protein was
measured according to the previously described by Bradford
method.61–64 and bovine serum albumin was used as the stand-
ard.65,66 After the purification method of the CA isoforms, samples
were subjected to SDS polyacrylamide gel electrophoresis (SDS-
PAGE).67–69 The change in activity was spectrophotometrically
obtained at 348 nm.70,71 The IC50 values were calculated from
activity (%) against compounds inhibition.72–74 Three various con-
centrations were used to calculate Ki values.75–77
The goal of this paper is to design and synthesize some novel
aminomethyl and alkoxymethyl derivatives (1–17) and to generate
more potent BChE and AChE enzymes, CA II and I isoforms.
Experimental
Chemistry
Synthesis of aminomethyl derivatives of benzothiazole and ben-
zoxazolthiones (1–10)
Aminomethylation was carried out at the temperature of 10ꢁC by
adding the corresponding aminal to a solution of mercaptobenzo-
thiazole (or mercaptobenzoxazole) in ethanol. The resulting prod-
uct was recrystallized from methanol. The aminomethyl derivatives
of benzothiazole and benzoxazolthiones 2–8 were reported in the
literature.47–53 However, there is no information about the synthe-
sis of compounds 9 and 10 in the literature.
Initial aminals were obtained by condensing of secondary
amines with formaldehyde. The physico-chemical characteristics of
the obtained products are shown in Table 1.
Formaldehyde was used as a form of paraformaldehyde. The
reaction was carried out in an absolute ethanol solution.
Hemiformals reacted immediately after its preparation without iso-
lation. The resulting reaction water was separated by azeotropic
distillation with benzene. The crystals were obtained after distilling
AChE/BChE activity determination and inhibition studies
The inhibitory effects of novel aminomethyl and alkoxymethyl
derivatives (1–17) on AChE and BChE activities were measured
according to Ellman et al.78 Acetylthiocholine iodide (AChI) and
butyrylthiocholine iodide (BChI) were used as substrates for the
reaction. 5,50-Dithio-bis(2-nitro-benzoic)acid (DTNB) was used for
the measurement of the AChE/BChE activities. Briefly, 1.0 ml of
the solvents, including ethanol and benzene, and recrystallizing. Tris/HCl buffer (1.0 M, pH 8.0), and 10 mL of sample solution were
The melting points and yields are given in Table 2.
dissolved in deionized water at different concentrations and 50 mL
AChE/BChE solution were mixed and incubated for 10 min at
25 ꢁC. Next 50 mL of DTNB (0.5 mM) was added. The reaction was
then initiated by the addition of 50 mL of AChI or BChI. The
hydrolysis of these substrates was monitored spectrophotometric-
ally by the formation of the yellow 5-thio-2-nitrobenzoate anion,
Synthesis of the alkoxymethyl derivatives of benzoxazolthione and
2-aminothiazole (11–17)
To do this, hemiformal was obtained from 0.05 mol of a formalde-
hyde (used as paraformaldehyde) and 40 ml of the corresponding as a result of the reaction of DTNB with thiocholine, which
alkanol (taken in excess as a solvent). Hemiformal reacted immedi- released by enzymatic hydrolysis of AChI or BChI, with absorption
maximum at 412 nm.
ately after its preparation without isolation. Then, 0.05 mol of

_
1176
I. GULC¸IN ET AL.
Table 1. Physico-chemical characteristics of aminomethyl derivatives of benzothiazol- and benzoxazolthiones
Found/calculated (%)
The melting Yield
No
Compound
point (ꢁC)
(%)
C
H
N
S
Brutto formula
C12H14N2OS2
NMR spectra d (ppm)
1
120–121
45
54.61 5.76 10.60 24.20
54.13 5.26 10.52 24.06
1.79 (kv. 2H, CH2CH2CH2), 3.29(t., 2H, NCH2), 3.9
(t., 2H, ŽCH2), 4.567 (t., 2H, NCH2N), 5.3
(s., 2H, NCH2O), 6.8–7.6 (m., 4H, C6H4).
N
O
CH2
N
N
S
S
S
2
124–126
40
56.00 5.90 10.30 26.60
57.60 5.30 10.50 24.60
C11H12N2OS2
1.819 (kv. 2H, CH2CH2CH2), 3.119 (t., 2H, NCH2),
3.9 (t., 2H, ŽCH2), 4.567 (t., 2H, NCH2N),
5.3 s. 2H (NCH2O), 7.1–7.3 m. 4H (C6H4)
CH
2N
O
S
3
4
134
40
42
54.9
5.55 11.01 24.56
C12H14N2OS2
C13H16N2S2
1.7 (m., 2H, CH2CH2CH2), 1.97 (m., 4H,
CH2CH2CH2), 3.01 (t., 4H, NCH2N), 7.6–8.9
(d., 4H, C5-C7), 8.1 (s., 2H, ArCH2N).
CH
N
2 N
O
54.13 5.26 10.52 24.06
S
S
152–152.5
57.6
57.0
7.5
5.6
11.3
11.2
28.8
25.6
1.5 (m., 2H, CH2CH2CH2), 1.7 (m., 4H,
CH2CH2CH2), 3.15 (t., 4H, NCH2N), 7.1–7.6
(d., 4H, C5-C7), 8 (s., 2H, ArCH2N).
N
CH2
N
N
N
S
S
O
S
5
6
7
128–130
105
35
65
45
54.4
5.98
8.2
27.0
25.4
C12H16N2S2
C12H16N2OS
C13H16N2OS
2.19 (kv, 2H, CH2CH2CH2), 3.12 (t., 2H, NCH2),
3.6 (t., 2H, ŽCH2), 4.67 (t., 2H, NCH2N), 5.20 s.
2H (NCH2O), 7.1–7.5 m. 4H (C6H4).
CH
2 N(C2H5)2
57.14 6.35 11.11
S
59.87 5.98 10.85 13.03
61.02 6.78 11.86 13.56
1.28 (m., 2H, CH2CH2CH2), 3.04 (t., 2H, CH2N),
3.69 t., (2H, CH2Ž), 4.59 (s., 2H, NCH2N),
5.36 (s., 2H, NCH2O), 6.9–7.9 (m., 4H, C6H4).
CH
2 N(C2H5)2
S
125
63.52 6.04
62.9
9.39
6.45 11.29
11.53
12.9
1.88 (m., 2H, CH2CH2CH2), 3.18 (t., 2H, CH2N),
3.9 (t., 2H, CH2Ž), 4.39 (s., 2H, NCH2N), 5.56
(s. 2H, NCH2O), 7.4–7.7 (m., 4H, C6H4).
CH
N
N
2 N
S
O
O
8
9
145–147
115–118
48
60
57.58
57.6
5.7
5.6
11.34 12.54
11.2 12.8
C12H14N2O2S
C11H12N2O2S
1.21 (d., 2H, CH2CH2CH2), 3.23 (t., 2H, CH2N),
3.7 (t., 2H, CH2Ž), 4.39 (s., 2H, NCH2N), 5.86
(s., 2H, NCH2O), 7.14–7.97 (m., 4H, C6H4).
CH
2 N
O
S
55.84 5.25 11.65 13.87
55.93 5.08 11.86 13.55
1.38 (m., 2H, CH2CH2CH2), 2.08 (t. 2H, CH2N),
3.39 (t., 2H, CH2Ž), 3.59 (s., 2H, NCH2N),
5.16 (s., 2H, NCH2O), 7.1–8.7 (m., 4H, C6H4).
N
O
CH2
N
N
S
O
O
10
145–147
67.6
56.0
57.6
5.51
5.6
11.0
11.2
13.3
12.8
C12H14N2O2S
1.8 (m., 2H, CH2CH2CH2), 3.08 (t., 2H, CH2N),
3.9 (t., 2H, CH2Ž), 4.59 (s., 2H, NCH2N), 5.56
(s.,. 2H, NCH2O), 7.4–7.7 (m., 4H, C6H4).
N
O
CH2
S
Results and discussion
Synthesis
Many physiologically active natural compounds contain > N-CH2-
O- > N-CH2-N < structural fragments. This study sought to build a
structure that combines physiologically active benzothiazole or
benzoxazole groups with alkoxymethyl or aminomethyl fragments.
Therefore, the aminomethylation and alkoxymethylation reactions
of mercaptobenzothiazole, mercaptobenzoxazole and 2-aminothia-
zole are developed.
The structure of the products was established by NMR spectros-
copy and the composition was confirmed by elemental analysis.
Spectra were measured on a Bruker device in acetone. A singlet at
4.6 ppm corresponding to N-CH2-N was observed in the 1H NMR
spectra of all aminomethyl derivatives. A singlet at 5.2–5.8 ppm
characterized the presence of the fragment N-CH2-O in the 1H
NMR spectra of alkoxymethyl derivatives. Methylene-bis-amines,
The alkoxymethyl derivatives of mercaptobenzoxazole and
which have good alkylation (amino-methylation) properties, were 2-aminothiazole were synthesized by reacting them with hemifor-
used as amino-methylation reagents. mals, which were prepared by the reaction of alcohols

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
1177
Table 2. Physico-chemical characteristics of the alkoxymethyl derivatives of benzoxazolthione and 2-aminothiazoles.
Found/calculated (%)
Melting
Brutto
Formula
No
Compounds
point (ꢁC)
Yield (%)
71.72
C
H
N
S
NMR spectra d (ppm)
11
130
55.40
55.38
4.58
4.61
7.13
7.18
16.35
16.41
C9H9NO2S
2.06 (s., 3H, OCH3); 5.781 (s., 2H,
NCH2O); 7.373–7.55 (m., 4H, C6H6).
CH
N
2OCH3
S
O
12
13
132–133
77
24
57.37
57.42
5.15
5.26
6.64
6.70
15.23
15.31
C10H11NO2S
2.06–2.096 (t., 3H, CH3); 3.07 (kv,
2H,CH2CH3); 5.766 (s., 2H, NCH2O);
7.33–7.52 (m., 4H, C6H4).
OC H
CH2
N
2
5
S
O
H
126–127
57.33
57.42
5.63
5.83
6.13
6.28
14.18
14.35
C11H13NO2S
2.01–2.16 (d., 6H, (CH3)2; 2.999
(m., 1H, OCH); 5.77 (s., 2H,
NCH2O); 7.34–7.53 m., 4H, C6H4).
O C(CH3)2
CH2
N
S
O
14
15
120–121
120–121
19
30
55.17
55.23
5.35
5.44
5.75
5.86
13.28
13.39
C11H13NO3S
C7H12N2SO2
1.06-2.01 (t., 3H, CH3), 3.072
(kv, 2H,CH2CH3), 5.62 (s., 2H,
NCH2O), 6.83–7.92 (m., 4H, C6H4).
OCH CH OCH
3
CH2
N
2
2
S
O
N
43.52
44.68
6.28
6.38
14.78
14.89
17.15
17.02
2.86 (s., 1H, NH); 5.11
(t., 4H,-OCH2CH2O-);
5.29 s. (3H, -OCH3); 5.20 (s., 2H,
NCH2O); 6.86 (d., 1H, SCH); 7.61
(d., 1H, NCH).
S
H
N
O CH CH OCH
CH2
2
2
3
16
17
118–120
126–127
–
47.98
48.84
6.35
6.98
15.97
16.28
17.80
18.6
C7H12N2SO
C5H8N2SO
1.11-2.10 (t., 3H, CH3), 3.10 (kv,
2H,CH2CH3), 5.20 (s., 2H, NCH2O),
6.83–7.792 (m., 4H, C6H4).
S
H
H
N
N
C(CH )
O
CH2
3
2
27
40.52
41.67
5.03
5.56
18.37
19.44
23.12
22.22
2.86–2.47 (s., 1H, NH); 5.19 (t., 4H,
-OCH2CH2O-); 5.19 s. (3H, -OCH3);
5.12 (s., 2H, NCH2O); 6.68 (d., 1H,
SCH); 7.71 (d., 1H, NCH).
S
H
N
N
CH2 OCH3
with formaldehyde.
antitumor activity) or hCA II (for antiglaucoma action) have been
developed.79,80 hCA II enhances sodium bicarbonate secretion in
the anterior uvea of the eye causing glaucoma and visual dysfunc-
tion.81 Heterocyclic molecules with primitive sulfonamide com-
pounds are the most extensively evaluated class of CAIs, which
has led to the advancement of diverse classes of clinical drugs like
methazolamide (MZA), acetazolamide (AZA) and others.82 In this
work, both the Ki and IC50 of the aminomethyl and alkoxymethyl
derivatives (1–17) were calculated and they are given in Table 3.
ROH þ CH2O ! ROCH2OH
1. Cytosolic hCA I, and II isoenzymes are widely distributed
throughout the human body and interference with these
enzymes may cause side effects. For the cytosolic hCA I
enzyme, aminomethyl and alkoxymethyl derivatives (1–17)
had Ki values in the range of 58 15 to 157 38 nM (Table 3).
Especially, compound 8 (Ki: 58 15 nM); N-morfolinomethyl-
benzoxazoline-2-thion and compound
5 (Ki: 70 15 nM);
N-diethylaminomethylbenzothiazoline-2-thione) inhibited the
hCA I isoform more potently than the standard compound
AZA (Ki: 333 28 nM), which is used to treat glaucoma, cysti-
nuria, periodic paralysis, epileptic seizure, dural estasia and
central sleep apnea. hCA I is involved in retinal edema and
cerebral and the inhibition of hCA I can be a significant factor
for eliminating of these conditions.83
2. The role of hCA II in diseases such as glaucoma has been well
characterized. Indeed, HCOꢀ3 production serves as a mechan-
ism to transport sodium ions (Naþ) into the eye along with
the influx of water, which leads to an increase in intraocular
pressure.84 Inhibition of CA II decreases HCOꢀ3 production and
Biological results
Sulfamate and sulfonamide CAIs demonstrated fundamental anti-
glaucoma and anti-tumour activities in vivo and in vitro; therefore
new therapeutic approaches targeting either hCA IX/XII (for

_
1178
I. GULC¸IN ET AL.
Table 3. AChE, human carbonic anhydrase I, and II isoforms (hCA I, and II) AChE and BChE enzymes inhibition effects of aminomethyl and alkoxymethyl derivatives
(1–17) and proportion of AChE to BChE enzymes.
IC50 (nM)
Ki (nM)
Compounds
hCA I
r2
hCA II
r2
AChE
r2
BChE
r2
hCA I
hCA II
AChE
BChE
AChE/BChE
1
2
3
4
5
6
7
8
79
79
83
86
82
102
79
94
103
98
112
119
105
112
128
156
142
373
—
0.9639
0.9852
0.9597
0.9588
0.9755
0.9359
0.9597
0.9533
0.9652
0.9550
0.9607
0.9752
0.9664
0.9426
0.9783
0.9757
0.9774
0.9774
—
104
114
89
99
98
0.9527
0.9839
0.9555
0.9774
0.9670
0.9649
0.9457
0.9619
0.9440
0.9452
0.9711
0.9695
0.9483
0.9556
0.9644
0.9659
0.9562
0.9816
—
51
39
54
36
52
44
65
38
62
50
89
63
63
38
43
76
80
—
174
0.9762
0.9885
0.9670
0.9855
0.9699
0.9857
0.9874
0.9848
0.9769
0.9819
0.9865
0.9908
0.9859
0.9860
0.9888
0.9949
0.9912
—
88
89
75
82
83
80
79
48
84
94
133
93
83
68
0.9964
0.9773
0.9619
0.9630
0.9359
0.9750
0.9520
0.9911
0.9904
0.9710
0.9621
0.9947
0.9807
0.9704
0.9752
0.9590
0.9749
—
108 35
77 15
106 39
86 40
70 15
123 48
82 17
58 15
76 20
99 22
118 40
95 25
90 23
119 53
118 27
132 30
157 38
333 28
—
81 19
107 11
96 22
26
33
37 11
18
34
45 11
40
32
78 36
61
45
75
34
23
25
42
5
5
32
51
45
41 13
51
30
57 17
23
80
50 10
77 17
88 11
7
4
9
0.812
0.647
0.822
0.439
0.666
1.500
0.701
1.391
0.975
1.220
0.584
0.852
0.971
0.383
0.555
0.763
1.285
—
105 25
100 24
115 28
132 27
121 40
135 33
135 42
137 35
146 46
94 37
142 56
193 79
162 29
215 40
353 60
—
2
9
9
3
118
116
119
121
137
172
141
122
128
158
167
187
520
—
8
6
3
9
9
10
11
12
13
14
15
16
17
AZAa
TACb
9
5
8
5
4
3
4
35 4
60 13
45 14
55 12
42 20
—
109
127
144
—
280
54 10
—
109
0.9513
0.9879
5
128 16
0.851
Tacrine (TAC) was used as a standard inhibitor for BChE and AChE enzymes.
aAcetazolamide (AZA) was used as a standard inhibitor for both carbonic anhydrase I, and II isoenzymes (hCA I and II).
subsequently aqueous humor secretion, which leads to 2-(methoxy)methylaminothiazole (17) compounds are weaker
decreased pressure in the eye.85 For the ubiquitous cytosolic inhibitors compared to other compounds for this isoenzyme. The
isoform hCA II, novel aminomethyl and alkoxymethyl deriva- molecule 8 was shown to had the excellent inhibitory efficacy on
tives (1–17) had Ki values ranging from 81 19–215 40 nM. hCA I isoenzyme activity while the molecule 1 was shown to had
In addition, AZA compound applied as a standard CA inhibi- the excellent inhibitory efficacy on hCA II isoenzyme activity. For
tor, which obtained Ki value of 353 60 nM. As can be hCA II isoform, the best inhibitors of them were N-oxazinomethyl-
observed in hCA II, the most considerable inhibition result benzothiazoline-2-thione (1) and N-isopropoxymethylbenzoxazo-
was recorded by N-oxazinomethylbenzothiazoline-2-thione (1) line-2-thione (13). The 2-(methoxyethoxy) methylaminothiazole
(81 19) (Table 3).
(15) and 2-methoxymethylaminothiazole (17) molecules are
weaker inhibitors compare with other molecules for this isoform.
As seen in Table 3 and Figure 2(b), IC50 values are in the range of
89–187 nM towards hCA II, while for hCA I is in the range of
79–156 nM. The IC50 values for standard molecule AZA towards
3. BChE and AChE were very significantly inhibited by novel
aminomethyl and alkoxymethyl derivatives (1–17). It was cal-
culated that Ki values were in the range of 23 3–88 11 nM
for BChE and 18 2–78 36 nM for AChE, respectively (Table
3). Additionally, tacrine (TAC) was used as clinically BChE and hCA II and I are 520 and 373 nM, respectively. All molecules have
AChE inhibitor, which had Ki values of 128 16 and lower IC50 value compare with AZA toward hCA II and hCA I
109 5 nM, respectively. The results are shown that entire of isoenzymes.
test molecules have perfect inhibition activity against BChE
and AChE compared to TAC.
As seen in Table 3 and Figure 2(c), IC50 amounts were in the
range of 36–89 nM towards AChE, while they were in the range of
4. In this work, we calculated AChE/BChE selectivity. The most 48–145 nM towards BChE (Figure 2(d)). The IC50 amounts of the
promising compound 14 obtained 2.22-fold of inhibitory standard compound TAC towards AChE and BChE were 174 and
activity against AChE/BChE than that of TAC. It can be as a 280 nM, respectively. Entire compounds have lower IC50 amount
potential factor for the therapy of AD. Also, as is shown in than TAC toward AChE and BChE. ChEIs have shown excellent effi-
Table 3, the compound 14 (N-(methoxyethoxy)methyl-benzox- cacy than placebo in clinical tests and are extensively prescribed
azoline-2-thione) showed the highest selectivity for AChE over as symptomatic therapy to ameliorate behavior and recognition in
BChE (ratio: 0.388) and weakest compound was 6 (N-diethyl- AD patients with moderate dementia27,28. TAC (9-Amino-1,2,3,4-
aminomethylbenzoxazoline-2-thione) (ratio 1:500).
tetrahydroacridine) compound is a reversible inhibitor of BChE and
AChE and the first drug to be agreed by the Drugs and
Foods Administration of America for the placative therapy of AD.35
For AChE and BChE enzymes were good inhibited by entire of
the evaluated compounds, the best inhibitors of AChE were
N-Piperidinomethylbenzothiazoline-2-thione (4), N-(methoxyethox-
y)methyl-benzoxazoline-2-thione (14) and also for BChE were
N-diethylaminomethylbenzoxazoline-2-thione (6) and N-morfolino-
methylbenzoxazoline-2-thion (8), respectively.
Discussion
The synthesized molecules are shown to inhibit hCA II and I isoen-
zymes by the interplay of aminomethyl and alkoxymethyl deriva-
tives (1–17) with cofactor Zn2þ ions in the structure of the
isoforms. For hCA I isoform (generally defined an important iso-
form when CAIs for anticancer activity or antiglaucoma are
encountered) was good inhibited by entire of the evaluated mole-
cules, the best inhibitors of them were N-diethylaminomethylben-
zothiazoline-2-thione (5), N-morfolinomethylbenzoxazoline-2-thion
Conclusions
(8)
and
N-oxazolidinomethylbenzoxazoline-2-thione
(9) In this paper, nanomolar levels of Ki amounts were obtained for
(Figure 2(a)). The 2-isopropoxymethylaminothiazole (16) and entire novel aminomethyl and alkoxymethyl derivatives (1–17) and

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
1179
(a)
(b)
300
600
400
200
0
AChE
BChE
hCA I
hCA II
200
100
0
AZA
2
4
6
8
10 12 14 16
TAC
3
6
9
12
15
Compounds
Compounds
(c)
(d)
400
300
200
100
0
150
100
50
AChE
BChE
hCA I
hCA II
0
AZA
2
4
6
8
10 12 14 16
TAC
2
4
6
8
10 12 14 16
Compounds
Compounds
Figure 2. (a) IC50 values of aminomethyl and alkoxymethyl derivatives for hCA I, and II isoenzymes. (b) IC50 values of aminomethyl and alkoxymethyl derivatives for
AChE and BChE enzymes. (c) Ki values of aminomethyl and alkoxymethyl derivatives for hCA I, and II isoenzymes. (d) Ki values of aminomethyl and alkoxymethyl deriva-
tives for AChE and BChE enzymes.
these molecules can be considerable inhibitor of AChE, BChE
enzymes and both hCA isoforms. The molecules 5 and 8 towards
hCA I and molecules 1 and 13 towards hCA II and molecules 4
and 14 towards AChE and molecules 6 and 8 towards BChE
enzymes recorded which can to be the leader molecules of the
parts for subsequent evaluations.
aryl)acryloyl) phenyl)-3a,4,7,7a-tetrahydro-1H-4,7-methanoi-
soindole-1,3(2H)-dione derivatives. Bioorg Chem 2017;70:
118–25.
3. Gokcen T, Al M, Topal M, et al. Synthesis of some natural
sulphonamide derivatives as carbonic anhydrase inhibitors.
Org Commun 2017;10:15–23.
€
4. Karalı N, Akdemir A, Goktas F, et al. Novel sulfonamide-con-
taining 2-indolinones that selectively inhibit tumor-associ-
ated alpha carbonic anhydrases. Bioorg Med Chem 2017;
25:3714–8.
Acknowledgements
S. Alwasel would like to thank the Distinguished Scientist
Fellowship Program, King Saud University for their support.
5. Aksu K, Ozgeris B, Taslimi P, Naderi A, et al. Antioxidant
activity, acetylcholinesterase, and carbonic anhydrase inhibi-
tory properties of novel ureas derived from phenethyl-
amines. Arch. Pharm. (Weinheim) 2016; 349:944–54.
Disclosure statement
_
6. Topal F, Gulcin I, Dastan A, Guney M. Novel eugenol deriva-
tives: potent acetylcholinesterase and carbonic anhydrase
inhibitors. Int J Biol Macromol 2017;94:845–51.
The authors declare no conflict of interest.
7. Ceylan M, Kocyigit UM, Usta NC, et al. Synthesis, carbonic
anhydrase I and II isoenzymes inhibition properties and anti-
bacterial activities of novel tetralone based 1,4-benzothiaze-
pine derivatives. J Biochem Mol Toxicol 2017;31:e21872.
8. Del Prete S, Vullo D, Osman SM, et al. Sulfonamide inhibition
profiles of the b-carbonic anhydrase from the pathogenic
bacterium Francisella tularensis responsible of the febrile ill-
ness tularemia. Bioorg Med Chem 2017;25:3555–61.
ORCID
_
Ilhami Gulc¸in
http://orcid.org/0000-0001-5993-1668
References
1. Cressier D, Procullac C, Hernande P, et al. Synthesis, antioxi-
dant properties and radioprotective effects of new benzo-
thiazoles and thiadiazoles. Bioorg Med Chem 2009;17:
5275–84.
9. Oktay K, Polat Kose L, Sendil K, et al. Synthesis of 3-chloro-1-
substituted aryl pyrrolidine-2,5-dione derivatives: discovery
of potent human carbonic anhydrase inhibitors. Med Chem
Res 2017;26:1619–27.
10. Bayrak C¸, Taslimi P, Gulc¸in I, Menzek A. The first synthesis
_
€
2. Kocyigit UM, Budak Y, Gurdere MB, et al. Synthesis, charac-
€
terization, anticancer, antimicrobial and carbonic anhydrase
inhibition profiles of novel (3aR,4S,7R,7aS)-2-(4-((E)-3-(3-
of 4-phenylbutenone derivative bromophenols including
natural products and their inhibition profiles for carbonic

_
1180
I. GULC¸IN ET AL.
€
anhydrase, acetylcholinesterase and butyrylcholinesterase 27. Oztas¸kın N, C¸etinkaya Y, Taslimi P, Goksu S, et al.
Antioxidant and acetylcholinesterase inhibition properties of
enzymes. Bioorg Chem 2017;72:359–66.
novel bromophenol derivatives. Bioorg Chem 2015;60:49–57.
11. Supuran CT, Capasso C. The g-class carbonic anhydrases as
drug targets for antimalarial agents. Expert Opin Ther
Targets 2015;19:551–63.
12. Akocak S, Lolak N, Nocentini A, et al. Synthesis and bio-
logical evaluation of novel aromatic and heterocyclic bis-sul-
fonamide Schiff bases as carbonic anhydrase I, II, VII and IX
inhibitors. Bioorg Med Chem 2017;25:3093–7.
_
€
€
28. Polat Kose L, Gu€lc¸in I, Goren AC, et al. LC-MS/MS analysis,
antioxidant and anticholinergic properties of galanga
(Alpinia officinarum Hance) rhizomes. Ind Crops Prod 2015;
74:712–21.
29. Scozzafava A, Kalın P, Supuran CT, et al. The impact of
hydroquinone on acetylcholine esterase and certain human
carbonic anhydrase isoenzymes (hCA I, II, IX, and XII).
J Enzyme Inhib Med Chem 2015;30:941–6.
30. Topal M, Gocer H, Topal F, et al. Antioxidant, antiradical and
anticholinergic properties of cynarin purified from the illyrian
thistle (Onopordum illyricum L.). J Enzyme Inhib Med Chem
2016;31:266–75.
_
ꢀ
13. Koc¸yigit UM, Aslan OM, Gu€lc¸in I, et al. inhibition of novel
2-(4-(aryl)thiazole-2-yl)-3a,4,7,7a-tetrahydro-1h-4,7-methanoi-
soindole-1,3(2h)-dione derivatives. Arch Pharm 2016;349:
955–63.
14. Akbaba Y, Bastem E, Topal F, et al. Synthesis and carbonic
anhydrase inhibitory effects of novel sulfamides derived
from 1-aminoindanes and anilines. Arch. Pharm. (Weinheim)
2014;347:950–7.
15. Innocenti A, Gulcin I, Scozzafava A, Supuran CT. Carbonic
anhydrase inhibitors. Antioxidant polyphenol natural prod-
ucts effectively inhibit mammalian isoforms I-XV. Bioorg Med
Chem Lett 2010;20:5050–3.
31. Gocer H, Topal F, Topal M, et al. Acetylcholinesterase and
carbonic anhydrase isoenzymes I and II inhibition profiles of
taxifolin. J Enzyme Inhib Med Chem 2016;31:441–7.
€
€
€
32. Ozgeris¸ B, Goksu S, Kose Polat L, et al. Acetylcholinesterase
and carbonic anhydrase inhibitory properties of novel urea
and sulfamide derivatives incorporating dopaminergic 2-ami-
notetralin scaffolds. Bioorg Med Chem 2016;24:2318–29.
_
€
€
16. S¸enturk M, Gulc¸in I, Beydemir S¸, et al. In vitro inhibition of
human carbonic anhydrase I and II isozymes with natural
phenolic compounds. Chem Biol Drugs Des 2011;77:494–9.
_
€
33. Gulc¸in I, Scozzafava A, Supuran CT, et al. The effect of caffeic
acid phenethyl ester (CAPE) metabolic enzymes including
acetylcholinesterase,
butyrylcholinesterase,
glutathione
_
€
17. C¸oban TA, Beydemir S, Gulc¸in I, Ekinci D. Morphine inhibits
erythrocyte carbonic anhydrase in vitro and in vivo. Biol
Pharm Bull 2007;30:2257–61.
18. Coban TA, Beydemir S, Gulc¸in I, Ekinci D. The effect of etha-
nol on erythrocyte carbonic anhydrase isoenzymes activity:
an in vitro and in vivo study. J Enzyme Inhib Med Chem
2008;23:266–70.
s-transferase, lactoperoxidase and carbonic anhydrase
ısoenzymes I, II, IX and XII. J Enzyme Inhib Med Chem 2016;
31:1095–101.
_
€
€
34. Yılmaz S, Akbaba Y, Ozgeris¸ B, et al. Synthesis and inhibitory
properties of some carbamates on carbonic anhydrase
and acetylcholine esterase. J Enzyme Inhib Med Chem
2016;31:1484–91.
19. Topal F, Topal M, Gocer H, et al. Antioxidant activity of taxi-
folin: an activity-structure relationship. J Enzyme Inhib Med
Chem 2016;31:674–83.
35. Sujayev A, Garibov E, Taslimi P, et al. Synthesis of some tet-
rahydropyrimidine-5-carboxylates, determination of their
metal chelating effects and inhibition profiles against acetyl-
cholinesterase, butyrylcholinesterase and carbonic anhy-
drase. J Enzyme Inhib Med Chem 2016;31:1531–9.
€
_
€
€
20. Innocenti A, Ozturk Sarıkaya SB, Gulc¸in I, Supuran CT.
Carbonic anhydrase inhibitors. Inhibition of mammalian iso-
forms I-XIV with a series of natural product polyphenols and
phenolic acids. Bioorg Med Chem 2010;18:2159–64.
€
ꢀ
36. Gul HI, Tugrak M, Sakagami H, et al. Synthesis and bioactiv-
ity studies on new 4-(3-(4-substitutedphenyl)-3a,4-dihydro-
3h-indeno[1,2-c]pyrazol-2-yl) benzenesulfonamides. J Enzyme
Inhib Med Chem 2016;31:1619–24.
_
€
€
21. S¸enturk M, Gulc¸in I, Das¸tan A, et al. Carbonic anhydrase
inhibitors. Inhibition of human erythrocyte isozymes I and II
with a series of antioxidant phenols. Bioorg Med Chem
2009;17:3207–11.
_
€
37. Gulc¸in I, Scozzafava A, Supuran CT, et al. Rosmarinic acid
inhibits some metabolic enzymes including glutathione S-
transferase, lactoperoxidase, acetylcholinesterase, butyrylcho-
linesterase, and carbonic anhydrase isoenzymes. J Enzyme
Inhib Med Chem 2016;31:1698–702.
€
€
ꢀ
22. Goc¸er H, Akıncıoglu A, Oztas¸kın N, et al. Synthesis, antioxi-
dant, and antiacetylcholinesterase activities of sulfonamide
derivatives of dopamine-related compounds. Arch. Pharm.
(Weinheim) 2013;346:783–92.
€
38. Genc¸ H, Kalin R, Koksal Z, et al. Discovery of potent carbonic
anhydrase and acetylcholinesterase inhibitors: 2-aminoindan
b-lactam derivatives. Int J Mol Sci 2016;17:1736.
39. Koc¸ak R, Turan A, E, Kalin P, et al. Synthesis of some novel
norbornene-fused pyridazines as potent inhibitors of car-
bonic anhydrase and acetylcholinesterase. J Heterocyc Chem
2016;53:2049–56.
40. Turan B, Sendil K, Sengul E, et al. The synthesis of some
b-lactams and investigation of their metal chelating activity,
carbonic anhydrase and achetylcholinesterase inhibition pro-
files. J Enzyme Inhib Med Chem 2016;31(Suppl 1):79–88.
_
ꢀ
€
€
23. Akıncıoglu A, Topal M, Gulc¸in I, Goksu S. Novel sulfamides
and sulfonamides incorporating tetralin scaffold as carbonic
anhydrase and acetylcholine esterase inhibitors. Arch Pharm
2014;347:68–76.
€
ꢀ
€
24. Goc¸er H, Akıncıoglu A, Goksu S, et al. Carbonic anhydrase
and acetylcholine esterase inhibitory effects of carbamates
and sulfamoylcarbamates. J Enzyme Inhib Med Chem 2015;
30:316–20.
€
25. Aksu K, Topal F, Gulc¸in I, et al. Acetylcholinesterase inhibi-
tory and antioxidant activities of novel symmetric sulfamides
derived from phenethylamines. Arch. Pharm. (Weinheim)
2015;348:446–55.
€
_
41. Ozbey F, Taslimi P, Gulcin I, et al. Synthesis, acetylcholin-
esterase, butyrilcholinesterase, carbonic anhydrase inhibitory
and metal chelating properties of some novel diaryl ether.
J Enzyme Inhib Med Chem 2016;31(Suppl 2):79–85.
ꢀ
ꢀ
€
26. Akıncıoglu A, Akıncıoglu H, Gulc¸in I, et al. Discovery of
potent carbonic anhydrase and acetylcholine esterase inhibi-
tors: Novel sulfamoylcarbamates and sulfamides derived
from acetophenones. Bioorg Med Chem 2015;23:3592–602.
42. Is¸ık M, Beydemir S, Yılmaz A, et al. Oxidative stress and
mRNA expression of acetylcholinesterase in the leukocytes
of ischemic patients. Biomed Pharmacother 2017;87:561–7.

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
1181
_
€
ꢀ €
€
43. Gul HI, Demirtas A, Ucar G, et al. Synthesis of Mannich bases 64. Koksal E, Aggul AG, Bursal E, Gulc¸in I. Purification and char-
by two different methods and evaluation of their acetylcho-
line esterase and carbonic anhydrase inhibitory activities.
Lett Drug Des Discov 2017;14:573–80.
acterization of peroxidase from sweet gourd (Cucurbita
Moschata Lam. Poiret). Int J Food Propert 2012;15:1110–9.
65. S¸ıktar E, Ekinci D, S¸ıktar E, et al. Protective role of L-carnitine
supplementation against exhaustive exercise-induced oxida-
tive stress in rats. Eur J Pharmacol 2011;668:407–13.
66. Aksu K, Nar M, Tanc¸ M, et al. The synthesis of sulfamide ana-
logues of dopamine related compounds and their carbonic
anhydrase inhibitory properties. Bioorg Med Chem 2013;
21:2925–31.
44. Woreka F, Schilha M, Neumaier K, et al. On-site analysis of
acetylcholinesterase and butyrylcholinesterase activity with
the ChE check mobile test kit: determination of reference
values and their relevance for diagnosis of exposure to
organophosphorus compounds. Toxicol Lett 2016;249:22–8.
45. Chen X, Zheng X, Zhou Z, et al. Effects of a cocaine
hydrolase engineered from human butyrylcholinesterase on
metabolic profile of cocaine in rats. Chem Biol Interact
2016;259:104–9.
46. Taslimi P, Sujayev A, Mamedova S, et al. Synthesis and bio-
activity of several new hetaryl sulfonamides. J Enzyme Inhib
Med Chem 2017;32:137–45.
47. Sorokin VG. Izvestiya Vysshikh Uchebnykh Zavedenii.
Khimiya I Khimicheskaya Tekhnologiya 1975;18:74–6.
48. Franklin S, Tamilvendan D, Venkatesa Prabhu G,
Balasubramanian T. Structural and spectral analysis of a
Mannich Base: 3-(Morpholin-4-ylmethyl)-1,3-benzothiazole-2-
thione. J Chem Crystallog 2012;42:29–33.
ꢀ
€
67. Akbaba Y, Akıncıoglu A, Goc¸er H, et al. Carbonic anhydrase
inhibitory properties of novel sulfonamide derivatives of
aminoindanes and aminotetralins. J Enzyme Inhib Med
Chem 2014;29:35–42.
_
€
€
68. C¸etinkaya Y, Goc¸er H, Gulc¸in I, Menzek A. Synthesis and car-
bonic anhydrase isoenzymes inhibitory effects of brominated
diphenylmethanone and its derivatives. Arch Pharm
(Weinheim) 2014;347:354–9.
_
€
69. Topal M, Gulc¸in I. Rosmarinic acid: a potent carbonic
anhydrase isoenzymes inhibitor. Turk
J Chem 2014;38:
894–902.
€
70. Guney M, Cos¸kun A, Topal F, et al. Oxidation of cyanobenzo-
cycloheptatrienes: synthesis, photooxygenation reaction and
carbonic anhydrase isoenzymes inhibition properties of
some new benzotropone derivatives. Bioorg Med Chem
2014;22:3537–43.
49. Hatayama, Kazuya Jpn. Kokai Tokkyo Koho. 1995; JP
07041604 A 19950210.
50. Valiuliene S, Kuodis Z, Rutavicius A, Chemija. 1994;(2):81-5n.
51. Dhal PN, Nayak A. Ind J Pharm 1975;37:92–4.
52. Yamaguchi J, Washisu S, Jpn. Kokai Tokkyo Koho. 1989; JP
01017048 A 19890120.
€
71. Goksu S, Naderi A, Akbaba Y, et al. Carbonic anhydrase
inhibitory properties of novel benzylsulfamides using
molecular modeling and experimental studies. Bioorg Chem
2014;56:75–82.
53. Erdogan B, Seyhan E, Atay O, Isikdag I. Gazi Universitesi
Eczacilik Fakultesi Dergisi 1989;6:163–72.
€
€
€
54. Ozturk Sarıkaya SB, Topal F, S¸enturk M, et al. In vitro inhib-
ition of a-carbonic anhydrase isozymes by some phenolic
compounds. Bioorg Med Chem Lett 2011;21:4259–62.
_
€
72. Arabaci B, Gulc¸in I, Alwasel S. Capsaicin: a potent inhibitor
of carbonic anhydrase isoenzymes. Molecules 2014;19:
10103–14.
_
€
55. Nar M, C¸etinkaya Y, Gulc¸in I, Menzek A. (3,4-
Dihydroxyphenyl)(2,3,4-trihydroxyphenyl)methanone and its
derivatives as carbonic anhydrase isoenzymes inhibitors. J
Enzyme Inhib Med Chem 2013;28:402–6.
73. Ozturk Sarikaya SB, Sisecioglu M, Cankaya M, et al.
Inhibition profile of a series of phenolic acids on bovine lac-
toperoxidase enzyme. J Enzyme Inhib Med Chem 2015;
30:479–83.
74. Boztas¸ M, C¸etinkaya Y, Topal M, et al. Synthesis and carbonic
anhydrase isoenzymes I, II, IX, and XII inhibitory effects of
dimethoxy-bromophenol derivatives incorporating cyclopro-
pane moieties. J Med Chem 2015;58:640–50.
75. Yıldırım A, Atmaca U, Keskin A, et al. N-Acylsulfonamides
strongly inhibit human carbonic anhydrase isoenzymes I and
II. Bioorg Med Chem 2015;23:2598–605.
76. Scozzafava A, Passaponti M, Supuran CT, et al. Carbonic
anhydrase inhibitors: Guaiacol and catechol derivatives
effectively inhibit certain human carbonic anhydrase isoen-
zymes (hCA I, II, IX, and XII). J Enzyme Inhib Med Chem
2015;30:586–91.
56. Verpoorte JA, Metha S, Edsall JT. Esterase activities of human
carbonic anhydrases B and C. J Biol Chem 1967;242:4221–9.
_
€
57. Coban TA, Beydemir S, Gucin I, et al. Sildenafil is a strong
activator of mammalian carbonic anhydrase isoforms I-XIV.
Bioorg Med Chem 2009;17:5791–5.
58. Ozturk Sarıkaya SB, Gulcin I, Supuran CT. Carbonic anhydrase
inhibitors. Inhibition of human erythrocyte isozymes I and II
with a series of phenolic acids. Chem Biol Drug Des
2010;75:515–20.
€
59. Artunc¸ T, C¸etinkaya Y, Goc¸er H, et al. Synthesis of 4-[2-(3,4-
dimethoxybenzyl)cyclopentyl]-1,2-dimethoxybenzene deriva-
tives and evaluations of their carbonic anhydrase isoen-
zymes inhibitory effects. Chem Biol Drug Des 2016;87:
594–607.
€
77. Aydin B, Gulcin I, Alwasel SH. Purification and characteriza-
tion of polyphenol oxidase from Hems¸in apple (Malus com-
munis L.). Int J Food Propert 2015;18:2735–45.
ꢀ
€
60. Akıncıoglu A, Akbaba Y, Goc¸er H, et al. Novel sulfamides as
potential carbonic anhydrase isoenzymes inhibitors. Bioorg
Med Chem 2013;21:1379–85.
78. Ellman GL, Courtney KD, Andres V, Featherston RM. A new
and rapid colorimetric determination of acetylcholinesterase
activity. Biochem Pharmacol 1961;7:88–95.
61. Bradford MM. A rapid and sensitive method for the quantita-
tion of microgram quantities of protein utilizing the prin-
ciple of protein-dye binding. Anal Biochem 1976;72:248–51.
_
€
79. Gocer H, Aslan A, Gulc¸in I, Supuran CT. Spirobisnaphthalenes
effectively inhibit carbonic anhydrase. J Enzyme Inhib Med
Chem 2016;31:503–7.
€
_
€_
€
€
ꢀ
62. Atasaver A, Ozdemir H, Gulc¸in I, Kufrevioglu OI. One-step
purification of lactoperoxidase from bovine milk by affinity
chromatography. Food Chem 2013;136:864–70.
_
€ €
80. Kuc¸uk M, Gulcin I. Purification and characterization of car-
bonic anhydrase enzyme from black sea trout (Salmo trutta
Labrax Coruhensis) kidney and inhibition effects of some
metal ions on the enzyme activity. Environ Toxicol
Pharmacol 2016;44:134–9.
_
€
ꢀ
€
63. S¸is¸ecioglu M, Gulc¸in I, C¸ankaya M, Ozdemir H. The inhibitory
effects of L-Adrenaline on lactoperoxidase enzyme (LPO)
purified from buffalo milk. Int J Food Propert 2012;15:
1182–9.

_
1182
I. GULC¸IN ET AL.
81. Go€ksu H, Topal M, Keskin A, et al. 9,10-Dibromo-N-aryl-9,10-
human carbonic anhydrase
I and II inhibitory effects.
dihydro-9,10-[3,4]epipyrroloanthracene-12,14-diones: synthesis
J Enzyme Inhib Med Chem 2016;31:1192–7.
and investigation of their effects on carbonic anhydrase iso- 84. Supuran CT. Structure and function of carbonic anhydrases.
zymes I, II, IX, and XII. Arch Pharm 2016;349:466–74. Biochem J 2016;473:2023–32.
82. Polat Kose L, Gulc¸in I, Ozdemir H, et al. The effects of 85. Rogez-Florent T, Foulona C, Drucbert AS, et al. Chiral separ-
_
€
€
some avermectins on bovine carbonic anhydrase enzyme.
J Enzyme Inhib Med Chem 2016;31:773–8.
83. Sujayev A, Polat Kose L, Garibov E, et al. Synthesis of N-alkyl
(aril)-tetra pyrimidine thiones and investigation of their
ation of new sulfonamide derivatives and evaluation of their
enantioselective affinity for human carbonic anhydrase II by
microscale thermophoresis and surface plasmon resonance.
J Pharm Biomed Anal 2017;137:113–22.