Synthesis of nitrogen, phosphorus, selenium and sulfur-containing heterocyclic compounds – Determination of their carbonic anhydrase, acetylcholinesterase, butyrylcholinesterase and α-glycosidase inhibition properties

Article abstract of DOI:10.1016/j.bioorg.2020.104171

Sulfur-containing pyrroles (1–3), tris(2-pyridyl)phosphine(selenide) sulfide (4–5) and 4-benzyl-6-(thiophen-2-yl)pyrimidin-2-amine (6) were synthesized and characterized by elemental analysis, IR and NMR spectra. In this study, the synthesized compounds o

Full text of DOI:10.1016/j.bioorg.2020.104171

BioorganicChemistry103(2020)104171
Contents lists available at ScienceDirect
Bioorganic Chemistry
journal homepage: www.elsevier.com/locate/bioorg
Synthesis of nitrogen, phosphorus, selenium and sulfur-containing
heterocyclic compounds – Determination of their carbonic anhydrase,
acetylcholinesterase, butyrylcholinesterase and α-glycosidase inhibition
properties
⁎
İlhami Gülçina, , Boris Trofimovb, Ruya Kayaa,c, Parham Taslimid, Lyubov Sobeninab,
Elena Schmidtb, Olga Petrovab, Svetlana Malyshevab, Nina Gusarovab, Vagif Farzaliyeve,
Afsun Sujayeve, Saleh Alwaself, Claudiu T. Supurang,h
a Atatürk University, Faculty of Sciences, Department of Chemistry, 25240 Erzurum, Turkey
b Irkutsk Institute of Chemistry of the Siberian Branch of the Russian Academy of Sciences, 664033 Irkutsk, Russia
c Central Research and Application Laboratory, Agri Ibrahim Cecen University, 04100 Agri, Turkey
d Department of Biotechnology, Faculty of Science, Bartin University, 74100 Bartin, Turkey
e Institute of Chemistry of Additives, Azerbaijan National Academy of Sciences, 1029 Baku, Azerbaijan
f Department of Zoology, College of Science, King Saud University, Riyadh, Saudi Arabia
g Dipartimento di Chimica Ugo Schiff, Universita degli Studi di Firenze, Sesto Fiorentino, Firenze, Italy
h Neurofarba Department and Laboratorio di Chimica Bioinorganica Universita‘ degli Studi di Firenze, Sesto Fiorentino, Italy
A R T I C L E I N F O
A B S T R A C T
Keywords:
Sulfur-containing pyrroles (1–3), tris(2-pyridyl)phosphine(selenide) sulfide (4–5) and 4-benzyl-6-(thiophen-2-
yl)pyrimidin-2-amine (6) were synthesized and characterized by elemental analysis, IR and NMR spectra. In this
study, the synthesized compounds of nitrogen, phosphorus, selenium and sulfur-containing heterocyclic com-
pounds (1–6) were evaluated against the human erythrocyte carbonic anhydrase I, and II isoenzymes, acet-
ylcholinesterase (AChE), butyrylcholinesterase (BChE), and α-glycosidase enzymes. The synthesized heterocyclic
compounds showed IC50 values in range of 33.32–60.79 nM against hCA I, and 37.05–66.64 nM against hCA II
closely associated with various physiological and pathological processes. On the other hand, IC50 values were
found in range of 13.13–22.21 nM against AChE, 0.54–31.22 nM against BChE, and 13.51–26.55 nM against α-
glycosidase as a hydrolytic enzyme. As a result, nitrogen, phosphorus, selenium and sulfur-containing hetero-
cyclic compounds (1–6) demonstrated potent inhibition profiles against indicated metabolic enzymes. Therefore,
we believe that these results may contribute to the development of new drugs particularly in the treatment of
some global disorders including glaucoma, Alzheimer’s disease and diabetes.
Pyrrole
Carbonic anhydrase
Enzyme inhibition
α-Glycosidase
Acetylcholinesterase, Butyrylcholinesterase
1. Introduction
The electron-rich nitrogen heterocycles, for instance pyrroles, pyr-
idines, pyrimidines, are not only able to readily accept or donate a
Pyrroles are widely used in medicinal chemistry and drug discovery.
Pyrrole derivatives are found in varieties of biological context as part of
co-factors and natural products. Common naturally produced molecules
containing pyrrole nucleus include Vitamin B12, bile pigments like
bilirubin and biliverdin, porphyrins of haeme, chlorophyll, chlorines,
bacteriochlorines and porphyrinogens [1]. The attention to the func-
tionalized pyrroles is now growing because some of them exhibit a large
spectrum of bioactivities including carbonic anhydrase isoenzyme in-
hibition effects [2–4].
proton, but they can also easily establish diverse weak interactions.
Some of these intermolecular processes such as like hydrogen bonding
formation, dipole-dipole interactions, hydrophobic effects, van der
Waals forces and π-stacking interactions of nitrogen compounds have
increased their importance in the field of medicinal chemistry and al-
lows them to bind with a variety of enzymes and receptors in biological
targets with high affinity due to their improved solubility [5].
Pyrrole-containing analogs are considered as a potential source of
biologically active compounds that contains a significant set of
⁎ Corresponding author at: Department of Chemistry, Faculty of Science, Ataturk University, 25240 Erzurum, Turkey.
E-mail address: igulcin@atauni.edu.tr (İ. Gülçin).
https://doi.org/10.1016/j.bioorg.2020.104171
Received 7 April 2020; Received in revised form 13 July 2020; Accepted 25 August 2020
Availableonline26August2020
0045-2068/©2020ElsevierInc.Allrightsreserved.

İ. Gülçin, et al.
BioorganicChemistry103(2020)104171
advantageous properties and can be found in many natural products.
The marketed drugs containing a pyrrole ring system are known to have
many biological properties such as antipsychotic, β-adrenergic an-
tagonist, anxiolytic, anticancer, antibacterial, antifungal, antiprotozoal,
antimalarial and many more [6a]. Pyrrole derivatives, pyrrolizines,
constitute a class of pyrrolic compound, which can serve as promising
scaffolds for anticancer drugs. For instance, mitomycin C has antitumor
activities against a wide variety of cancer types [6b]. Pyrroles deco-
rated with sulfur-containing functionalities are a rarity. Pyrroles
bearing thioester functions (S-alkyl, R1C(O)SR2) are amongst the best
known, courtesy of the critical role that thioesterase-appended pyrroles
play in the biosynthesis of secondary metabolites containing pyrroles
[7a,7b].
delayed or blocked the digestion mechanisms of carbohydrates, but
does not cause malabsorption [57–59].
In the present work, we have studied a series of recently synthe-
sized, earlier unknown nitrogen, sulfur, selenium and phosphorus
polyfunctional compounds, including pyrrolecarbodithioates, amino-
pyrimidine, tris(2-pyridyl)phosphinesulfide and –selenide as potential
functional biological active compounds. Mechanism of their action was
investigated and relationship between structure and efficiency of their
biological activity was estimated. Especially, in the light of this in-
formation, the aim of this study was to investigate the inhibition im-
pacts of nitrogen, phosphorus, selenium and sulfur-containing hetero-
cyclic compounds (1–6) on hCA I, and hCA II isoenzymes, AChE, BChE,
and α-glycosidase enzymes. Also, their inhibition profiles were com-
pared to the standard compounds like acetazolamide, tacrine and
acarbose as a clinical used inhibitors.
Carbonic anhydrases (CAs, E.C.4.2.1.1) are zinc-containing me-
talloenzymes. They catalyzes the reversibly conversion of carbon di-
oxide (CO2) and water to bicarbonate (HCO3−) and proton (H+) ions
[8–10]. In the mammal, when CO2 in the blood plasma passes into red
blood cells through diffusion, it is rapidly converted into carbonic acid
with the CA enzyme. CAs are found in both eukaryotic and prokaryotic
cells and encoded by eight distinct gene families including α-, β-, γ-, δ-,
ζ-, η-, θ-, and t-CAs [11–13]. However, only the α-CA family is found in
mammals [14–16]. The human carbonic anhydrases (hCAs) are be-
longing to the α-subfamily of CAs. This subfamily divided into sixteen
diverse isoforms. They have different catalytic activity, amino acid se-
quence, kinetic properties, cellular and tissue distributions, biochemical
properties, subcellular localization and sensitivity to inhibitors or ac-
tivators [17–19]. α-CAs are grouped as cytoplasmic isoenzymes in-
cluding CAs I, II, III, VII and XIII, membrane bound CAs including IV,
IX, XII, XIV and XV. CA IV is a glycosylphosphatidylinositol (GPI)-an-
chored protein. CA V is mitochondrial isoenzyme. CA VI is secretory CA
isoform. Lastly, CAs VIII, X and XI are CA-related proteins (CARPs).
However, CARPs have not performed CO2 hydration activity and phy-
siological function [20–22]. CA inhibitors (CAIs) show numerous
bioactivities linked to many global diseases such as glaucoma, obesity,
cancer, epilepsy and osteoporosis [23–26]. Due to these important
physiological functions, numerous studies have been performed on CAs.
Both CA I, and CA II are the most studied and examined isoenzymes
[27–29]. CA I is expressed in erythrocytes and the gastrointestinal tract,
on the other hand, CA II is expressed in almost all tissues mainly in-
cluding eye, erythrocytes, gastrointestinal tract, bone osteoclasts,
kidney, brain, testis and lung [30,31]. CA II isoenzyme has a single
chain protein including 295 amino acids with nearly 29 kDa molecular
weight [32–34]. Despite significant and increasing improvements in
CAIs, there are many difficulties such as high levels of isoenzymes in
tissues and CA inhibitors selectivity [35–37].
2. Material and methods
2.1. Chemistry
2.1.1. General information
IR spectra were recorded in KBr pellets or film on a Bruker JFS-25
spectrometer in the 400–4000 cm−1 region. 1H, 13C, 15N, 31P and 77Se
NMR spectra were recorded at room temperature on a Bruker-DPX-400
instrument with an operating frequency of 400.13 (1H), 100.62 (13C),
40.5 (15N), 162.0 (31P), 77.0 (77Se) MHz, the solvent is DMSO‑d6 and
1
CDCl3. H and 13C NMR chemical shifts were recorded relative to re-
sidual solvent signals (CDCl3, δ 7.27 and 77.0 ppm, respectively, or
DMSO, δ 2.50 and 39.5 ppm, respectively). Elemental analysis was
performed on a Flash EA 1112 Series analyzer.
2.1.2. Ethyl 5-phenyl-1H-pyrrole-2-carbodithioate (1)
A mixture of 2-phenylpyrrole (1.43 g, 10 mmol) and KOH·5H2O
(1.30 g, 20 mmol) in DMSO (20 mL) is stirred for 30 min and carbon
disulfide (1.52 g, 20 mmol) is added. The reaction mixture is allowed to
stand at room temperature for 2 h and then ethyl iodide (1.56 g,
10 mmol) is added. The reaction mixture is stirred for 2 h, then diluted
with water (40 mL) and extracted with diethyl ether. After removing
the solvent the residue is passed through column (Al2O3, hexane-diethyl
ether, 1:1) to afford: 0.2 g of diethyl disulfide; 0.7 g (24%) of ethyl 5-
phenyl-1H-pyrrole-1-carbodithioate and 1.46 g (59%) of target ethyl 5-
phenyl-1H-pyrrole-2-carbodithioate as yellow crystals, mp 67–68 °C. 1H
NMR (400 MHz, DMSO‑d6): 11.79 (br s, 1H, NH), 7.93–7.91 (m, 2H, Ho
Ph), 7.44–7.42 (m, 2H, Hm Ph), 7.34–7.33 (m, 1H, Hp Ph), 7.19 (dd,
J = 3.6, 2.4 Hz, H-3 pyrrole), 6.79 (dd, J = 3.6, 2.6 Hz, H-4 pyrrole),
3.33 (q, J = 7.5 Hz, 2H, SCH2), 1.30 (t, J = 7.5 Hz, Me). 13C NMR
(101 MHz, DMSO‑d6): 206.0 (C]S), 141.8 (C-i), 140.5 (C-5), 130.8 (C-
2), 129.0 (C-m), 128.6 (C-p), 126.3 (C-o), 115.0 (C-4), 110.6 (C-3), 28.6
(SCH2), 13.5 (Me).
Dementia is one of the fastest spreading diseases in the world.
Alzheimer’s disease (AD) leads to emotional trouble and escorted by
abnormality in cholinergic neurotransmission of the central nervous
system [38–41]. The pathophysiology of AD is relevant to the depletion
of the neurotransmitter acetylcholine (ACh), associated with AChE ac-
tivity. AChE (E.C.3.1.1.7) hydrolyzes the neurotransmitter ACh
[42–45]. Butyrylcholinesterase (BChE, E.C.3.1.1.8) created inside liver,
consider as a backup for the analogous AChE. Also, BChE had a crucial
role in the cholinergic mechanism, and its inhibition improved the
cognitive functions. Both cholinergic enzymes share some structural
similarities, containing the presence of a catalytic center. The searches
of new cholinesterase inhibitors sound a significant strategy to famil-
iarize new drug candidates against AD and related dementias [46–50].
Diabetes mellitus (DM) was known as a popular metabolic degen-
erative disorder, is characterized by a high level of blood glucose.
Antidiabetic treatments diagnose that determining blood glucose
amounts are effective therapy for diabetes [51–53]. α-Glycosidases are
found on the brushy surface of the small intestine and responsible for
the breakdown of complex carbohydrate polymers [54–56]. Also, they
divide disaccharides and oligosaccharides into monosaccharides, which
easily absorbed from the intestinal walls. α-Glycosidase inhibitor
2.1.3. 2-[(Ethylsulfanyl)(4,5,6,7-tetrahydro-1H-indol-2-yl)methylene]
malononitrile (2)
Malonodinitrile (0.99 g, 15 mmol), KOH·5H2O (0.98 g, 15 mmol)
and DMSO (50 mL) are stirred at room temperature for 0.5 h, then ethyl
4,5,6,7-tetyrahydroindole-carbodithioare (2.25 g, 10 mmol) is added
and the mixture is heated at 108–110 °C for 1.5 h. The reaction mixture
is diluted with water (1:3), the crystals formed are filtered off and dried
to afford 2.26 g (88%) of 2-[(ethylsulfanyl)(4,5,6,7-tetrahydro-1H-
indol-2-yl)methylene]malononitrile, mp 138–139 °C (methanol). 1H
(DMSO‑d6): 11.81 (br s, 1H, NH), 6.93 (d, J = 1.8 Hz, 1H, H-3), 3.05 (q,
J = 7.4 Hz, 2H, SCH2), 2.67–2.66 (m, 2H, CH2-7), 2.50–2.49 (m, 2H,
CH2-4), 1.74–1.73 (m, 2H, CH2-6), 1.69–1.68 (m, 2H, CH2-5), 1.18 (t,
J = 7.4 Hz, 3H, Me). 13C (CDCl3-d3): 159.3 (=C-SEt), 141.6 (C-5),
126.1 (C-2), 124.8 (C-4), 120.8 (C-3), 116.6 (CN), 115.5 (CN), 68.2
[=C(CN)2)], 31.2 (SCH2), 23.5 (CH2-7), 23.0 (CH2-5), 22.7 (CH2-6),
22.4 (CH2-4), 14.6 (SCH2Me).
2

İ. Gülçin, et al.
BioorganicChemistry103(2020)104171
2.1.4. 3-Imino-1-ethylsulfanyl-5,6,7,8-tetrahydro-3H-pyrrolo[1,2–a]
indole-2-carboxamide (3)
(10 mL) was heated (100 °C) and stirred at 100 °C for 30 min. After
cooling (70 °C), H2O (0.090 g, 5.0 mmol) and (NH2)2C = NH⋅HCl
(0.573 g, 6.0 mmol) were added to the reaction mixture and stirred at
70 °C for 0.5 h. Then KOH⋅0·.5H2O (0.325 mg, 5.0 mmol) was added
and the mixture was stirred at 70 °C for 30 min. The reaction mixture,
after cooling at room temperature, was diluted with H2O (15 mL),
neutralized with NH4Cl and was extracted with CHCl3 (10 mL × 4).
The organic extract was washed with H2O (5 mL × 3) and dried
(MgSO4). CHCl3 was evaporated in vacuum and the residue was pur-
ified by column chromatography (Al2O3, eluent C6H6/Et2O with gra-
dient from 1:0 to 10:1). 4-Benzyl-6-(thiophen-2-yl)pyrimidin-2-amine
was isolated as a cream solid (0.401 g, 30% yield); m.p. 180–185 °C;
elemental analysis calcd (%) for C15H13N3S (267.35): C 67.39%; H
4.90%; N 15.72%; S 11.99%; found: C 67.37%; H 4.91%; N 15.62%; S
11.94%. IR (film): νmax 3485, 3289, 3166, 3100, 3083, 3028, 2920,
1622, 1573, 1556, 1548, 1518, 1494, 1459, 1446, 1416, 1366, 1342,
1227, 1194, 1172, 1074, 1044, 1030, 908, 862, 839, 826, 791, 736,
728, 707, 622, 608, 577, 568, 520 cm−1; 1H NMR (400.1 MHz, CDCl3):
δ 3.93 (s, 2H, CH2Ph), 5.05 (br.s, 2H, NH2), 6.72 (s, 1H, H5), 7.06 (dd,
J = 3.8, 5.0 Hz, 1H, H9), 7.20–7.42 (m, 5H, Ho,m,p), 7.41 (d, J = 5.0 Hz,
1H, H10), 7.57 (d, J = 3.8 Hz, 1H, H8) ppm; 13C NMR (100.6 MHz,
CDCl3): δ 44.2 (CH2Ph), 105.4 (C5), 126.8 (Cp), 127.0 (C8), 128.1 (C9),
128.7 (Cm), 129.2 (C10), 129.3 (Co), 138.0 (Ci), 143.0 (C7), 160.3 (C6),
163.2 (C4), 170.9 (C2) ppm; 15N NMR (40.5 MHz, CDCl3): δ –136.6
(N3), –148.3 (N1), –305.6 (NH2) ppm.
Solution of 2-cyano-3-(ethylsulfanyl)-3-(4,5,6,7-tetrahydro-1H-
indol-2-yl)acrylamide (0.55 g, 2 mmol) in methanol (10 mL) is heated
in the presence of triethylamine (2–3 drops) for 10 h and cooled to
room temperature. The crystals formed are filtered off, washed with
diethyl ether and dried to afford 3-imino-1-ethylsulfanyl-5,6,7,8-tetra-
hydro-3H-pyrrolo[1,2–a]indole-2-carboxamide (0.25 g, 46%), mp
1
190–192 °C. H NMR (CDCl3-d3): 8.51 (br s, 1H, CONH2), 7.76 (br s,
1H, =NH), 6.16 (s, 1H, pyrrole), 5.42 (br s, 1H, CONH2), 3.19 (q,
J = 7.2 Hz, SCH2), 2.72–2.70 (m, 2H, CH2-7), 2.47–2.45 (m, 2H, CH2-
4), 1.90–1.88 (m, 2H, CH2-5), 1.78–1.76 (m, 2H, CH2-6), 1.43 (t,
J = 7.2 Hz, Me). 13C NMR (CDCl3-d3): 165.15 (CO), 157.79 (C-1),
155.58 (C-3), 131.21 (C-4), 129.45 (C-7), 125.93 (C-5), 113.91 (C-6),
112.76 (C-2), 26.2 (SCH2), 22.9, 22.7, 22.2, 22.1 (CH2-4–7), 14.3
(SCH2Me).
2.1.5. Tris(2-pyridyl)phosphine sulfide (4)
Tris(2-pyridyl)phosphine (0.521 g, 2 mmol) was dissolved in di-
chloromethane (10 mL) and elemental selenium was added (0.155 g,
2.01 mmol) at room temperature. The reaction mixture was stirred. The
reaction was completed in 2 h with higher yield. The solution was fil-
tered and solvent removed. The white crystalline product was crystal-
lized from ethanol, and dried on air. The purity of the tri(2-pyridyl)
phosphine selenide was checked by IR, 1H, 13C, 31P, 77Se NMR spectra
and melting point. Yield 95%, m.p. 175 °C; lit. 176–178 °C (EtOH).
Found: C, 52.3; H, 3.5; N, 12.5; P, 9.1; Se, 22.9%. Anal. Calcd. for:
2.2. Biochemical studies
C
15H12N3PSe. C, 52.3; H, 3.5; N, 12.2; P, 9.0; Se, 22.9%. 1H NMR
3
(400 MHz, CDCl3), δ ppm: 7.31 (dddd 3H, JHH = 7.8 Hz,
3JHH = 4.7 Hz, 3JHH = 3.1 Hz, 4JHH = 1.2 Hz, H5 Py), 7.79 tdd (3H, H4-
Py, 3JHH = 7.8 Hz, 3JHH = 4.7 Hz, 4JHH = 1.8 Hz), 8.30 ddd (3H, H3-
Py, 3JHH = 7.8 Hz, 3JHP = 6.8 Hz, 4JHH = 1.2 Hz), 8.69 br.d (3H, H6-
Py, 3JHH = 4.8 Hz). 13C NMR (100.62 MHz, CDCl3): δ = 124.88 (C5-Py,
2.2.1. Carbonic anhydrase purification studies
In this work, both hCA I, and II isoenzymes were purified
by Sepharose-4B-L-Tyrosine-sulfanilamide affinity chromatography
[60,61]. Sepharose-4B-L-Tyrosine-sulfanilamide was used as an affinity
matrix for selective retention of both hCA isoenzymes [62–64]. Both
hCA isoenzymes activity was spectrophotometrically determined ac-
cording to previous method described in details [65,66]. p-Ni-
trophenylacetate (PNA) was used as substrate and transformed to p-
nitrophenolate ions (PNP) [67]. One CA isoenzyme unit is accepted as
the amount of CA, which had absorbance change at 348 nm of PNA to
PNP over a period of 3 min at 25 °C [68,69].
4JPC = 3.2 Hz), 129.22 (C3-Py, JPC = 26.0 Hz), 136.17 (C4-Py,
2
3
3JPC = 10.5 Hz), 149.86 (C6-Py, JPC = 19.0 Hz), 154.12 (C2-Py,
1JPC = 106.0 Hz). 31P NMR (161.98 MHz, CDCl3): δ = 30.13 ppm,
(1JPSe = 732.6 Hz). 77Se NMR (77.0 MHz, CDCl3): δ = -307.2 ppm,
(1JPSe = 746.5 Hz). IR (KBr pellet, cm−1): 3063, 1566, 1449, 1420,
1282, 1124, 1050, 985, 772, 734, 560, 505, 450.
After affinity chromatography, the tubes containing the enzyme
were identified at 280 nm [70]. The protein quantity was spectro-
photometrically determined at 595 nm according to the Bradford
method [71]. During the purification steps, bovine serum albumin was
used as the standard protein [72]. Purity control of CA isozymes was
determined the two different acrylamides concentration. It was carried
out in 10 and 3% acrylamides for the running and the stacking gel,
respectively, containing 0.1% sodium dodecyl sulphate (SDS) according
to Laemmli procedure [73] as described in previous studies [74,75].
2.1.6. Tris(2-pyridyl)phosphine selenide (5)
Tris(2-pyridyl)phosphine (0.521 g, 2 mmol) was dissolved in di-
chloromethane (10 mL) and elemental sulfur was added (0.064 g,
2.01 mmol) at room temperature. The reaction mixture was stirred. The
reaction was completed in 2 h with higher yield. The solution was fil-
tered and solvent removed. The white crystalline product was crystal-
lized from isopropanol, and dried on air. The purity of the tri(2-pyridyl)
phosphine sulfide was checked by IR, 1H, 13C, 31P NMR spectra and
melting point. Yield 95%, m.p. 158–160 °C (isoPrOH); Yield 95%, lit.
160 °C (CHCl3) [2]. Found: C, 60.7; H, 4.0; N, 14.3; P, 19.1; S, 10.9%.
Anal. Calcd. for: C15H12N3PS. C, 60.6; H, 4.1; N, 14.1; P, 19.4; S, 10.8%.
2.2.2. Cholinesterases assays
AChE from electric eel (Electrophorus electricus) and BChE from
equine serum inhibitory effects of nitrogen, phosphorus, selenium and
sulfur-containing heterocyclic compounds (1–6) were measured by
slightly modifying the colorimetric Ellman’s method [76] as described
previously [77,78]. Acetylthiocholine iodide (AChI) and butyr-
ylthiocholine iodide (BChI) were used as substrates of the both enzy-
matic reactions. Additionally, 5,5-dithiobis(2-nitro-benzoic acid)
(DTNB) was used as common substrate for the determination of the
AChE and BChE activities. Briefly, 1 mL of Tris/HCl buffer (pH 8.0,
1.0 M) and 10 μL of sample solution at different concentrations were
dissolved in deionized water. Then, an aliquot AChE or BChE enzymes
(50 μL) was mixed and incubated at room temperature for 10 min. After
incubation period, an aliquot of DTNB (50 μL, 0.5 mM) was added.
Then, the reaction was allowed to start by the addition of 50 μL of AChI
or BChI (10 mM). The breakdown of these substrates was monitored
spectrophotometrically by yellow color formation of 5-thio-2-
1H NMR (400 MHz, CDCl3), δ ppm: 7.32 (dddd 3H, JHH = 7.8 Hz,
3
3JHH = 4.8 Hz, 4JHH = 1.2 Hz, 5JHH = 2.9 Hz, H5 Py), 7.77 tdd (3H, H4-
Py, 3JHH = 7.8 Hz, 4JPH = 4.6 Hz, 4JHH = 1.7 Hz), 8.25 ddd (3H, H3–
Py, 3JHH = 7.8 Hz, 3JHP = 6.5 Hz, 4JHH = 1.0 Hz), 8.70 br. d (3H, H6-
Py, 3JHH = 4.8 Hz). 13C NMR (100.62 MHz, CDCl3), δ: 124.90 (C5- Py,
2
4JPC = 3.3 Hz), 128.56 (C3– Py, JPC = 25.0 Hz), 136.04 (C4-Py,
3
3JPC = 10.2 Hz), 149.87 (C6-Py, JPC = 19.3 Hz), 155.05 (C2– Py,
1JPC = 114.6 Hz). 31P NMR (161.98 MHz, CDCl3): δ = 34.8 ppm. IR
(KBr pellet, cm−1): 3037, 2986, 1571, 1449, 1419, 1279, 1235, 1208,
1161, 1051, 1131, 1085, 1042, 987, 774, 742, 731, 661, 614, 549, 519,
473, 438, 394.
2.1.7. 4-Benzyl-6-(thiophen-2-yl)pyrimidin-2-amine (6)
A mixture of 2-acethylthiophene (0.631 g, 5.0 mmol), phenylace-
tylene (0.510 g, 5.0 mmol), and KOBut (0.673 g, 6.0 mmol) in DMSO
3

İ. Gülçin, et al.
BioorganicChemistry103(2020)104171
nitrobenzoate anion as the result of the reaction of DTNB with thio-
choline from hydrolysis of AChI or BChI with absorption at a wave-
length of 412 nm [79,80].
alkylthio groups, on the base of reaction of pyrroles with carbon dis-
ulfide (Scheme 1a) and transformations of pyrrole-2-carboditioates thus
formed in functionalized 2-vinylpyrroles with alkylthio groups
(Scheme 1b) or products of their intramolecular cyclization, 1-al-
kylthio-3-imino-3H-pyrrolizines (Scheme 1c) [89].
2.2.3. α-Glycosidase assay
5-Phenylpyrrole-2-carbodithioate (1) was chosen taking into ac-
count the known sensitivity of pyrroles to oxidizers, on the one hand,
and physiological properties, especially the antioxidant activity
of sulfide and thiocarbonyl fragments, on the other hand. For the same
The inhibitory effect of the nitrogen, phosphorus, selenium and
sulfur-containing heterocyclic compounds (1–6) on α-glycosidase en-
zyme, which obtained from Saccharomyces cerevisiae, was carried out
using the p-nitrophenyl-D-glycopyranoside (p-NPG) substrate according
to the analysis of Tao et al. [81] as described previously [82,83].
Briefly, 200 μL of phosphate buffer was mixed with 40 μL of homo-
genate solution in same buffer solution (0.15 EU/mL, pH 7.4). After
preincubation, 50 μL of p-NPG in phosphate buffer (5 mM, pH 7.4) was
added and incubated again at 30 °C. Absorbances were measured
spectrophotometrically at 405 nm according to previous studies [84].
reasons, pyrrole compounds
2
(2,2-dicyanoethenyl-4,5,6,7-tetra-
hydroindole) and 3 (3-imino-1-ethylsulfanyl-5,6,7,8-tetrahydro-3H-
pyrrolo[1,2–a]indole-2-carboxamide) were selected as containing di-
valent sulfur atoms, functional groups, and exo- and endocyclic double
bonds sensitive to radical attack. Tris(2-pyridyl)phosphine sulfide and
selenide was synthesized tris(2-pyridyl)phosphine and elemental sulfur
and selenium. Tris(2-pyridyl)phosphine was prepared according to the
our literature procedure from red phosphorus and 2-chloropyridine in
superbasic system KOH/DMSO (Scheme 2) [90]. Tris-(2-pyridyl)phos-
phine sulfide (4) and tris-(2-pyridyl)phosphine selenide (5) were sup-
posed to be antioxidants of combined action owing to the high ability of
oxidizing the P]S and P = Se bonds. In addition, the choice of these
compounds is due to the fact that they possess a high physiological
activity, suppressing various pathological processes occurring in living
organisms.
2.2.4. Determination of inhibitor parameter
Metabolic enzymes inhibition by the nitrogen, phosphorus, sele-
nium and sulfur-containing heterocyclic compounds (1–6) was mea-
sured for each substrate. Control activity was taken as 100%. The
sulfur-containing heterocyclic compounds concentrations causing 50%
inhibition (IC50) values were calculated from activity (%)-Nitrogen,
phosphorus, selenium and sulfur-containing heterocyclic compounds
concentration graph as described previously in details [85–87]. The Ki
values were determined from a series of experiments using three dif-
ferent nitrogen, phosphorus, selenium and sulfur-containing hetero-
cyclic compounds (1–6) and substrates at five different concentrations
to construct Lineweaver–Burk curves as described previously [88].
4-Benzyl-6-(thiophen-2-yl)pyrimidin-2-amine was prepared from 2-
acethylthiophene, phenylacetylene and (NH2)2C = NH⋅HCl [13]
(Scheme 2) [91]. At the same time It was expected that the thiophene-
aminopyrimidine ensemble (4-benzyl-6-(thiophen-2-yl)pyrimidin-2-
amine) (6) would be able to exhibit physiological properties due to its
potentially high and multifaceted chemical activity.
3. Results and discussion
CA isozymes are a broad family of enzymes involved the some
physiological and pathological processes. Especially, involvement of
physiologically relevant CA I, and II isoenzymes in many crucial phy-
siological and biological processes such as regulation of the acid-base
homeostasis made them act as worthy drug targets in epilepsy, glau-
coma, and cerebral edema [92]. So, recent advances in the use of CA
inhibitors as new antiobesity, anticancer and anti-infective drugs have
All compounds are synthesized by original straightforward methods
from available starting materials. When selecting the compounds for
the study, we consider that if some of them would show promising
performance characteristics, the synthetic procedures might be easily
scaled up. We have developed the methodology of synthesis of sulfur-
containing pyrroles, including dithioesters and compounds with
S
1.CS2, 2.Etl
KOH/DMSO
N
H
N
SEt
H
1
(a)
1.CS2, 2.Etl
KOH/DMSO
1.CH2(CN)X, 2.Etl
S
SEt
CN
KOH/DMSO
N
H
N
N
H
SEt
H
NC
2
(b)
SEt
(Et)3N
SEt
N
H
N
MeOH
CONH2
NC
CONH2
HN
(c)
3
Scheme 1. The synthesis route of compounds 1–3.
4

İ. Gülçin, et al.
BioorganicChemistry103(2020)104171
N
N
P
S
N
N
N
CH2Cl2
r.t.
KOH/DMSO
Pred
+
N
P
4
N
Cl
N
N
P
Se
N
(~
70%)
5
(
~
95%)
1. KOBut/DMSO
2. (NH2)2C=NH . HCl
3. KOH
CH3
N
+
One-pot
O
N
NH2
S
6
Scheme 2. The synthesis route of compounds 4–6.
Table 1
The half maximal inhibition concentration (IC50 values, nM) of some nitrogen, phosphorus, selenium and sulfur-containing heterocyclic compounds (1–6) against
carbonic anhydrase I, and II isoenzymes (hCA I and II), acetylcholinesterase (AChE) and α-glycosidase (α-Gly) enzymes.
Compounds
hCA I
r2
hCA II
r2
AChE
r2
BChE
r2
α-Gly
r2
1
48.46
45.0
59.74
33.32
56.34
60.79
25.80
–
0.9205
37.05
38.5
51.72
37.66
53.72
66.64
23.70
–
0.9609
0.9833
0.9424
0.9956
0.9352
0.9339
0.9825
–
13.13
17.34
18.09
14.44
19.8
22.21
–
0.9895
0.9752
0.9508
0.9739
0.9435
0.9316
–
27.18
28.40
27.07
24.93
31.22
0.54
–
0.9761
0.9611
0.9936
0.9810
0.9922
0.9383
–
26.55
19.85
17.91
13.51
18.88
24.06
–
0.9400
0.9555
0.9384
0.9375
0.9315
0.9575
–
2
93.03
0.9629
0.9459
0.9810
0.9424
0.9887
–
3
4
5
6
AZA*
TAC*
ACR*
5.57
–
0.9937
–
4.67
–
0.9579
–
–
–
–
–
–
–
22,800
–
*Acetazolamide (AZA) was used as a standard inhibitor for both hCA I, and II isoenzymes. Tacrine (TAC) was used as a standard inhibitor for AChE and BChE
enzymes. Acarbose (ACR) was used as a standard inhibitor for α-glycosidase and taken from reference of [81].
been widely emphasized in recent years, as well as diuretics and anti-
glaucoma medications [93,94]. In this study, we investigated the effects
of nitrogen, phosphorus, selenium and sulfur-containing heterocyclic
compounds (1–6) on hCA I, and II isoenzymes purified from human red
blood cells using by Sepharose-4B-L-Tyrosine affinity column chroma-
tography. Also, for controlling of both isoenzymes purities, SDS-poly-
acrylamide gel electrophoresis (PAGE) was performed [95] and single
band observed for each purified hCA isoenzyme. Purification results of
hCA I, and hCA II isoenzymes were summarized in Table 1. As for bot
isoenzymes, nitrogen, phosphorus, selenium and sulfur-containing
heterocyclic compounds (1–6) showed IC50 values in the low nano-
molar range. IC50 value refers to the binding affinity of the inhibitor to
the enzyme [96]. The CA I is a physiologically relevant isoenzyme,
found at the highest level in erythrocytes and is also expressed in
normal colorectal mucosa and encoded by the CA1 gene in humans
[97]. It is expressed in normal colorectal mucosa and exists at the
highest level in red blood cells [98]. Therefore, nitrogen, phosphorus,
selenium and sulfur-containing heterocyclic compounds (1–6) demon-
strated high affinity and low nanomolar inhibition levels against both
isoenzymes. IC50 values of nitrogen, phosphorus, selenium and sulfur-
containing heterocyclic compounds (1–6) were found between 33.32
and 60.79 nM for hCA I, and 37.05–66.64 nM for hCA II, which includes
the primary Na-carrying mechanism into the eyes and regulate the in-
traocular pressure (Table 1). Lover IC50 value indicates that the in-
hibitor is bound to the enzyme with strong affinity [99,100]. So, a close
look at the inhibitors of CA II is very important for treatment of glau-
coma [101,102]. Also, Among them, tris(2-pyridyl)phosphine selenide
(4) had strong inhibition effect against hCA I isoenzyme (IC50
:
33.32 nM, r2: 0.9459), and the physiologically relevant hCA II isoform
(IC50: 37.05 nM, r2: 0.9956). On the other hand, acetazolamide (AZA),
which used as the reference inhibitor, had IC50 values were found as
25.80 nM (r2: 0.9887) and 23.70 nM (r2: 0.9825) for hCA I and hCA II
isoenzymes, respectively. These results clearly showed that nitrogen,
phosphorus, selenium and sulfur-containing heterocyclic compounds
(1–6) had strong of similar inhibitory effect to AZA against both hCA
isoenzymes (Table 1). AZA, dorzolamide, methazolamide, di-
chlorphenamide, ethoxzolamide, and sulthiame are examples of CA
inhibitors that are clinically used for the treatment of glaucoma, obesity
5

İ. Gülçin, et al.
BioorganicChemistry103(2020)104171
and epilepsy [103]. It is well known CA II inhibitors are directly bound
to the zinc metal in the center of active site of hCA II. Generally, at the
bottom catalytic active site of both hCA is a cone-shaped cavity that is
partitioned into two conserved environments, hydrophilic and hydro-
phobic walls, with a Zn2+ ion. The Zn2+ ion is coordinated as tetra-
hedral with putative three histidine residues (His94, His96, and
His119) and a solvent molecule (H2O). It can bind to hydroxide ion
(eOH) followed by reaction with CO2 to yield HCO3– ions. In this form,
CAIs should include zinc-binding group, which can successfully co-
ordinate with the binding site Zn2+ ion. Zinc-binding group is coupled
with variable chemical moieties that known as tail. This tail facilitates
the interactions with specific amino acid residues within the active site
of CA. Although the CA binding site is highly conserved among the
different isoenzymes, there is variability in the polarity and hydro-
phobicity of its peripheral part [104].
4. Conclusions
In conclusion, the inhibitory effects of some nitrogen, phosphorus,
selenium and sulfur-containing heterocyclic compounds on hCA I, and
II isoenzymes and AChE, BChE and α-glycosidase enzymes were eval-
uated together. The sulfur-containing heterocyclic compounds we used
in our study showed inhibition effects on hCA I, and II isoenzymes and
AChE, BChE and α-glycosidase enzymes activities at low nanomolar
concentrations. We believe that these results may be useful in the
synthesis of new CA isoenzyme inhibitors and in the development of
drugs for the treatment of some diseases.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
AChE and BChE inhibition properties of novel nitrogen, phosphorus,
selenium and sulfur-containing heterocyclic compounds (1–6) were
determined according to the Ellman’s procedure [76] as previously
described [105]. Novel nitrogen, phosphorus, selenium and sulfur-
containing heterocyclic compounds (1–6) are the potent derivatives
with IC50 values in ranging between 13.13 and 22.21 nM for AChE
(Table 1) and 0.54 to 31.22 nM for BChE, whereas, TAC had IC50 value
of 5.57 nM against AChE (r2: 0.9937) and 5.57 nM against BChE (r2:
0.9579). All studied sulfur-containing heterocyclic compounds de-
monstrated powerful inhibition against cholinergic enzyme of AChE,
but ethyl 5-phenyl-1H-pyrrole-2-carbodithioate (1) showed the best
inhibition effect against AChE (IC50: 13.13 nM; r2: 0.9895) (Table 1). It
is well known that pyrrole and fused pyrrole compounds possess a wide
spectrum of pharmacological activities mainly acting as anticancer
agents [106]. On the other hand, when the inhibition results for BChE
were examined, it was clearly observed that 4-benzyl-6-(thiophen-2-yl)
pyrimidin-2-amine (6) was the best inhibitor with IC50 value of
0.54 nM, which 8.65 times greater than tacrine (IC50: 4.67 nM; r2:
0.9579). Compound 6 is the strongest against BChE. Based on these
results, it can be said that BChE is the more prevalent enzyme within
age and in AD, in detriment of AChE.
Acknowledgements
S.A would like to extend his sincere appreciation to the Researchers
Supporting Project (RSP-2019/59), King Saud University, Saudi Arabia.
Appendix A. Supplementary material
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.bioorg.2020.104171.
References
[1] S.S. Gholap, Pyrrole: An emerging scaffold for construction of valuable therapeutic
agents, Eur. J. Med. Chem. 110 (2016) 13–31.
[2] M.M. Ghorab, M. Ceruso, M.S. Alsaid, Y.M. Nissan, C.T. Supuran, Novel sulfona-
mides bearing pyrrole and pyrrolopyrimidine moieties as carbonic anhydrase in-
hibitors: Synthesis, cytotoxic activity and molecular modeling, Eur. J. Med. Chem.
87 (2014) 186–196.
[3] M.M. Ghorab, M.S. Alsaid, M. Ceruso, Y.M. Nissan, C.T. Supuran, Carbonic an-
hydrase inhibitors: Synthesis, molecular docking, cytotoxic and inhibition of the
human carbonic anhydrase isoforms I, II, IX, XII with novel benzenesulfonamides
incorporating pyrrole, pyrrolopyrimidine and fused pyrrolopyrimidine moieties,
Bioorg. Med. Chem. 22 (2014) 3684–3695.
Finally, α-glycosidase was much more inhibited by all tested ni-
trogen, phosphorus, selenium and sulfur-containing heterocyclic com-
pounds (1–6) (IC50: 13.51–26.55 nM) (Table 1). The results showed
that all nitrogen, phosphorus, selenium and sulfur-containing hetero-
cyclic compounds (1–6) had effective α-glycosidase inhibition effects
than that of acarbose (IC50: 22800 nM) as standard α-glycosidase in-
hibitor [81]. Also, when the results were evaluated, tris(2-pyridyl)
phosphine selenide (4) was found to be the best inhibitor for α-glyco-
sidase (IC50:13.51 nM; r2: 0.9375). In fact, nowadays, it is well estab-
lished that there is a connection between diabetes and AD; in fact, a
common pathophysiology between both diseases has been previously
suggested [107a]. In this study, there is epidemiological and basic
science evidence, which suggest a possible shared pathophysiology
between T2DM and AD. Also, it was hypothesized that AD might be
type 3 diabetes (T3DM), but definitive biochemical mechanisms remain
unknown [107a]. Currently, there is a rapid increasing studies pointing
toward insulin deficiency and insulin resistance as mediators of AD-
type neurodegeneration. However, this new fluctuation of information
is full of contradictory and unsolved concepts related to the potential
contributions of type T2DM, obesity, metabolic syndrome, and to AD
pathogenesis [107b].
[4] M.M. Ghorab, M.S. Alsaid, M. Ceruso, Y.M. Nissan, C.T. Supuran, Pyrrolo and
pyrrolopyrimidine sulfonamides act as cytotoxic agents in hypoxia via inhibition
of transmembrane carbonic anhydrases, Eur. J. Med. Chem. 188 (2020) 112021.
[5] N. Kerru, L. Gummidi, S. Maddila, K.K. Gangu, S.B. Jonnalagadda, A review on
recent advances in nitrogen-containing molecules and their biological applica-
tions, Molecules 25 (2020) 1909–1951.
[6] (a) V. Bhardwaj, D. Gumber, V. Abbot, S. Dhimana, P. Sharma, Pyrrole: a re-
sourceful small molecule in key medicinal hetero-aromatics, RSC Adv. 5 (2015)
15233–15266;
(b) A. Belal, B. El-Gendy, M. El-D, Pyrrolizines: Promising scaffolds for anticancer
drugs, Bioorg. Med. Chem. 22 (2014) 46–53.
[7] (a) B.R. Groves, D.A. Smithen, T.S. Cameron, A. Thompson, Thionation reactions
of 2-pyrrole carboxylates, RSC Adv. 6 (2016) 69691–69697;
(b) A.E. Gamal, V. Agarwal, S. Diethelm, I. Rahman, M.A. Schorn, J.M. Sneed,
G.V. Louie, K.E. Whalen, T.J. Mincer, J.P. Noel, V.J. Paul, B.S. Moore, Biosynthesis
of coral settlement cue tetrabromopyrrole in marine bacteria by a uniquely
adapted brominase-thioesterase enzyme pair, Proc. Natl. Acad. Sci. 113 (2016)
3797–3802.
[8] (a) C.T. Supuran, Carbonic anhydrases and metabolism, Metabolites 8 (2018) 25;
(b) Y. Cetinkaya, H. Göçer, I. Gulcin, A. Menzek, Synthesis and carbonic anhy-
drase isoenzymes inhibitory effects of brominated diphenylmethanone and its
derivatives, Arch. Pharm. 347 (5) (2014) 354–359;
(c) C. Bayrak, P. Taslimi, I. Gulcin, A. Menzek, The first synthesis of 4-phe-
nylbutenone derivative bromophenols including natural products and their in-
hibition profiles for carbonic anhydrase, acetylcholinesterase and butyr-
ylcholinesterase enzymes, Bioorg. Chem. 72 (2017) 359–366.
Brain diabetes has been associated with cognitive impairment and
alteration of acetylcholine as neurotransmitter responsible for cognitive
functionality. Amyloid formation is one of the most evident patholo-
gical events in both diseases in pancreas of diabetic patients, and Aβ in
the brain of Alzheimer’s patients [108a]. In fact, many authors claim
that antidiabetic agents (targeting, for instance, α-glucosidase) could be
useful for preventing or even treating AD [108b]. I think that correla-
tion between both diseases within the introduction section can improve
interest towards the family of compounds presented herein.
[9] (a) A. Karimov, A. Orujova, P. Taslimi, N. Sadeghian, B. Mammadov,
H.S. Karaman, V. Farzaliyev, E. Sujayev, R. Taş, S. Alwasel, I. Gulcin, Novel
functionally substituted esters based on sodium diethyldithiocarbamate deriva-
tives: synthesis, characterization, biological activity and molecular docking stu-
dies, Bioorg. Chem. 99 (2020) 103762;
(b) O. Hisar, Ş. Beydemir, I. Gulcin, Ö.İ. Küfrevioğlu, C.T. Supuran, Effect of low
molecular weight plasma inhibitors of rainbow trout (Oncorhyncytes mykiss) on
human erythrocytes carbonic anhydrase-II isozyme activity in vitro and rat ery-
throcytes in vivo, J. Enzyme Inhib. Med. Chem. 20 (2005) 35–39.
6

İ. Gülçin, et al.
BioorganicChemistry103(2020)104171
[10] (a) A. Aktas, D. Barut Celepci, Y. Gök, P. Taslimi, H. Akıncıoğlu, I. Gulcin, A novel
Ag-N-heterocyclic carbene complex bearing the hydroxyethyl ligand: synthesis,
characterization, crystal and spectral structures and bioactivity properties,
Crystals 10 (2020) 171;
[31] C. Caglayan, I. Gulcin, The toxicological effects of some avermectins on goat liver
carbonic anhydrase enzyme, J. Biochem. Mol. Toxicol. 32 (2018) e22010.
[32] I. Gulcin, M. Abbasova, P. Taslimi, Z. Huyut, L. Safarova, A. Sujayev, V. Farzaliyev,
S. Beydemir, S.H. Alwasel, C.T. Supuran, Synthesis and biological evaluation of
aminomethyl and alkoxymethyl derivatives as carbonic anhydrase, acet-
ylcholinesterase and butyrylcholinesterase inhibitors, J. Enzyme. Inhib. Med.
Chem. 32 (2017) 1174–1182.
(b) N. Oztaşkin, P. Taslimi, A. Maraş, S. Göksu, I. Gulcin, Novel antioxidant
bromophenols with acetylcholinesterase, butyrylcholinesterase and carbonic an-
hydrase inhibitory actions, Bioorg. Chem 74 (2017) 104–114.
[11] (a) S. Burmaoğlu, E. Akın Kazancioglu, R. Kaya, M. Kazancioglu, M. Karaman,
O. Algül, I. Gulcin, Synthesis of novel organohalogen chalcone derivatives and
screening of their molecular docking study and some enzymes inhibition effects, J.
Mol. Struct. 1208 (2020) 127868;
[33] A. Akıncıoğlu, E. Kocaman, H. Akıncıoğlu, R.E. Salmas, S. Durdağı, I. Gulcin,
C.T. Supuran, S. Göksu, The synthesis of novel sulfamides derived from β-ben-
zylphenethylamines as acetylcholinesterase, butyrylcholinesterase and carbonic
anhydrase enzymes inhibitors, Bioorg. Chem. 74 (2017) 238–250.
[34] H.I. Gul, C. Yamali, F. Yeşilyurt, H. Sakagami, K. Kucukoglu, I. Gulcin, M. Gül,
C.T. Supuran, Microwave assisted synthesis and bioevaluation of new sulfona-
mides, J. Enzyme Inhib. Med. Chem. 32 (2017) 369–374.
(b) H.I. Gül, M. Tuğrak, H. Sakagami, P. Taslimi, I. Gulcin, C.T. Supuran,
Synthesis and bioactivity studies on new 4-(3-(4-substitutedphenyl)-3a,4-dihydro-
3h-indeno[1,2-c]pyrazol-2-yl) benzenesulfonamides, J. Enzyme Inhib. Med.
Chem. 31 (6) (2016) 1619–1624.
[35] H.I. Gul, E. Mete, P. Taslimi, I. Gulcin, C.T. Supuran, Synthesis, carbonic anhy-
drase I and II inhibition studies of the 1,3,5-trisubstituted-pyrazolines, J. Enzyme
Inhib. Med. Chem. 32 (2017) 189–192.
[12] (a) C. Yamali, H.I. Gul, C. Kazaz, S. Levent, S. Levent, I. Gulcin, Synthesis,
structure elucidation, and in vitro pharmacological evaluation of novel polyfluoro
substituted pyrazoline type sulfonamides as multi-target agents for inhibition of
acetylcholinesterase and carbonic anhydrase I and II enzymes, Bioorg. Chem. 96
(2020) 103627;
[36] K. Kucukoglu, F. Oral, T. Aydin, C. Yamali, O. Algul, H. Sakagami, I. Gulcin,
C.T. Supuran, H.I. Gul, Synthesis, cytotoxicity and carbonic anhydrase inhibitory
activities of new pyrazolines, J. Enzyme Inhib. Med. Chem. 31 (S4) (2016) 20–24.
[37] E. Garibov, P. Taslimi, A. Sujayev, Z. Bingöl, S. Çetinkaya, I. Gulcin, S. Beydemir,
V. Farzaliyev, S.H. Alwasel, C.T. Supuran, Synthesis of 4,5-disubstituted-2-thioxo-
1,2,3,4-tetrahydropyrimidines and investigation of their acetylcholinesterase,
butyrylcholinesterase, carbonic anhydrase I/II inhibitory and antioxidant activ-
ities, J. Enzyme Inhib. Med. Chem. 31 (S3) (2016) 1–9.
(b) F. Ozbey, P. Taslimi, İ. Gulcin, A. Maraş, S. Goksu, C.T. Supuran, Synthesis,
acetylcholinesterase, butyrilcholinesterase, carbonic anhydrase inhibitory and
metal chelating properties of some novel diaryl ether, J. Enzyme Inhib. Med.
Chem. 31 (S2) (2016) 79–85.
[13] M. Yigit, B. Yigit, P. Taslimi, I. Ozdemir, M. Karaman, I. Gulcin, Novel amine-
functionalized benzimidazolium salts: Synthesis, characterization, bioactivity, and
molecular docking studies, J. Mol. Struct. 1207 (2020) 127802.
[14] A. Sujayev, P. Taslimi, R. Kaya, B. Safarov, L. Aliyeva, V. Farzaliyev, I. Gulcin,
Synthesis, Characterization and biological evaluation of N-substituted triazinane-
2-thiones and theoretical-experimental mechanism of condensation reaction,
Appl. Organomet. Chem. 34 (2020) e5329.
[38] P. Taslimi, I. Gulcin, Antioxidant and anticholinergic properties of olivetol, J. Food
Biochem. 42 (2018) e12516.
[39] N. Oztaşkın, Y. Çetinkaya, P. Taslimi, S. Göksu, I. Gulcin, Antioxidant and acet-
ylcholinesterase inhibition properties of novel bromophenol derivatives, Bioorg.
Chem. 60 (2015) 49–57.
[40] L. Polat Köse, I. Gulcin, A.C. Gören, J. Namiesnik, A.L. Martinez-Ayala,
S. Gorinstein, LC-MS/MS analysis, antioxidant and anticholinergic properties of
galanga (Alpinia officinarum Hance) rhizomes, Ind. Crops Prod. 74 (2015)
712–721.
[15] K. Buldurun, N. Turan, E. Bursal, A. Mantarcı, F. Turkan, P. Taslimi, I. Gulcin,
Synthesis, spectroscopic properties, crystal structures, antioxidant activities and
enzyme inhibition determination of Co(II) and Fe(II) complexes of Schiff base, Res.
Chem. Intermed. 46 (2020) 283–297.
[41] A. Scozzafava, P. Kalın, C.T. Supuran, I. Gulcin, S. Alwasel, The impact of hy-
droquinone on acetylcholine esterase and certain human carbonic anhydrase
isoenzymes (hCA I, II, IX, and XII), J. Enzyme Inhib. Med. Chem. 30 (2015)
941–946.
[16] S. Bilginer, H.I. Gul, F.S. Erdal, H. Sakagami, S. Levent, I. Gulcin, C.T. Supuran,
Synthesis, cytotoxicities, and carbonic anhydrase inhibition potential of 6-(3-aryl-
2-propenoyl)-2(3H)-benzoxazolones, J. Enzyme Inhib. Med. Chem. 34 (2019)
1722–1729.
[42] A. Akıncıoğlu, H. Akıncıoğlu, I. Gulcin, S. Durdağı, C.T. Supuran, S. Göksu,
Discovery of potent carbonic anhydrase and acetylcholine esterase inhibitors:
Novel sulfamoylcarbamates and sulfamides derived from acetophenones, Bioorg.
Med. Chem. 23 (2015) 3592–3602.
[17] S. Bal, R. Kaya, P. Taslimi, A. Aktaş, M. Karaman, I. Gulcin, Novel 2-methylimi-
dazolium salts: synthesis, characterization, molecular docking, and carbonic an-
hydrase and acetylcholinesterase inhibitory properties, Bioorg. Chem. 94 (2020)
103468.
[43] M. Topal, H. Gocer, F. Topal, P. Kalin, P. Polat Köse, I. Gulcin, K.C. Çakmak,
M. Küçük, L. Durmaz, A.C. Gören, S.H. Alwasel, Antioxidant, antiradical and an-
ticholinergic properties of cynarin purified from the illyrian thistle (Onopordum
illyricum L.), J. Enzyme Inhib. Med. Chem. 31 (2016) 266–275.
[18] S. Bilginer, B. Gonder, H.I. Gul, R. Kaya, I. Gulcin, B. Anil, C.T. Supuran, Novel
sulfonamides incorporating triazene moieties show powerful carbonic anhydrase I
and II inhibitory properties, J. Enzyme Inhib. Med. Chem. 35 (2020) 325–329.
[19] R.F. Georgea, S. Bua, C.T. Supuran, F.M. Awadallah, Synthesis of some N-aroyl-2-
oxindole benzenesulfonamide conjugates with carbonic anhydrase inhibitory ac-
tivity, Bioorg. Chem. 96 (2020) 103635.
[44] H. Gocer, F. Topal, M. Topal, M. Küçük, D. Teke, I. Gulcin, S.H. Alwasel,
C.T. Supuran, Acetylcholinesterase and carbonic anhydrase isoenzymes I and II
inhibition profiles of taxifolin, J. Enzyme Inhib. Med. Chem. 31 (2016) 441–447.
[45] B. Ozgeriş, S. Göksu, L. Köse Polat, I. Gulcin, R.E. Salmas, S. Durdagi, F. Tümer,
C.T. Supuran, Acetylcholinesterase and carbonic anhydrase inhibitory properties
of novel urea and sulfamide derivatives incorporating dopaminergic 2-aminote-
tralin scaffolds, Bioorg. Chem. 24 (2016) 2318–2329.
[20] M. Tuğrak, H.İ. Gül, K. Bandow, H. Sakagami, I. Gulcin, Y. Ozkay, C.T. Supuran,
Synthesis and biological evaluation of some new mono Mannich bases with pi-
perazines as possible anticancer agents and carbonic anhydrase inhibitors, Bioorg.
Chem. 90 (2019) 103095.
[21] H.I. Gül, M. Tuğrak, M. Gül, S. Mazlumoğlu, H. Sakagami, I. Gulcin, C.T. Supuran,
New phenolic mannich bases with piperazines and their bioactivities, Bioorg.
Chem. 90 (2019) 103057.
[46] I. Gulcin, A. Scozzafava, C.T. Supuran, H. Akıncıoğlu, Z. Koksal, F. Turkan,
S. Alwasel, The effect of caffeic acid phenethyl ester (CAPE) metabolic enzymes
including acetylcholinesterase, butyrylcholinesterase, glutathione S-transferase,
lactoperoxidase and carbonic anhydrase isoenzymes I, II, IX and XII, J. Enzyme
Inhib. Med. Chem. 31 (2016) 1095–1101.
[22] K. Küçükoğlu, H.İ. Gül, P. Taslimi, I. Gulcin, C.T. Supuran, Investigation of in-
hibitory properties of some hydrazone compounds on hCA I, hCA II and AChE
enzymes, Bioorg. Chem. 86 (2019) 316–321.
[47] S. Yılmaz, Y. Akbaba, B. Özgeriş, L. Polat Köse, S. Göksu, I. Gulcin, S.H. Alwasel,
C.T. Supuran, Synthesis and inhibitory properties of some carbamates on carbonic
anhydrase and acetylcholine esterase, J. Enzyme Inhib. Med. Chem. 31 (2016)
1484–1491.
[23] S. Burmaoğlu, A.O. Yılmaz, M.F. Polat, O. Algül, R. Kaya, I. Gulcin, Synthesis and
biological evaluation of novel tris-chalcones as potent carbonic anhydrase, acet-
ylcholinesterase, butyrylcholinesterase, and α-glycosidase inhibitors, Bioorg.
Chem. 85 (2019) 191–197.
[48] D. Ozmen Ozgun, C. Yamali, H.İ. Gül, P. Taslimi, I. Gulcin, T. Yanik, C.T. Supuran,
Inhibitory effects of isatin Mannich bases on carbonic anhydrases, acet-
ylcholinesterase and butyrylcholinesterase, J. Enzyme Inhib. Med. Chem. 31
(2016) 1498–1501.
[24] D. Ozmen Ozgün, H.İ. Gül, C. Yamali, H. Sakagami, I. Gulcin, M. Sukuroglu,
C.T. Supuran, Synthesis and bioactivities of pyrazoline benzensulfonamides as
carbonic anhydrase and acetylcholinesterase inhibitors with low cytotoxicity,
Bioorg. Chem. 84 (2019) 511–517.
[49] A. Sujayev, E. Garibov, P. Taslimi, I. Gulcin, S. Gojayeva, V. Farzaliyev,
S.H. Alwasel, C.T. Supuran, Synthesis of some tetrahydropyrimidine-5-carbox-
ylates, determination of their metal chelating effects and inhibition profiles
against acetylcholinesterase, butyrylcholinesterase and carbonic anhydrase, J.
Enzyme Inhib. Med. Chem. 31 (2016) 1531–1539.
[25] Ç. Bayrak, P. Taslimi, H.S. Kahraman, I. Gulcin, A. Menzek, The first synthesis,
carbonic anhydrase inhibition and anticholinergic activities of some bromophenol
derivatives with S including natural products, Bioorg. Chem. 85 (2019) 128–139.
[26] M. Tuğrak, H.İ. Gül, H. Sakagami, I. Gulcin, C.T. Supuran, New azafluorenones
with cytotoxic and carbonic anhydrase inhibitory properties: 2-Aryl-4-(4-hydro-
xyphenyl)-5H-indeno[1,2-b]pyridin-5-ones, Bioorg Chem. 81 (2019) 433–439.
[27] B. Yiğit, M. Yiğit, P. Taslimi, Y. Gok, I. Gulcin, Schiff bases and their amines:
synthesis, and discovery of carbonic anhydrase and acetylcholinesterase enzymes
inhibitors, Arch. Pharm. 351 (2018) e1800146U.
[50] I. Gulcin, A. Scozzafava, C.T. Supuran, Z. Koksal, F. Turkan, S. Çetinkaya,
Z. Bingöl, Z. Huyut, S.H. Alwasel, Rosmarinic acid inhibits some metabolic en-
zymes including glutathione S-transferase, lactoperoxidase, acetylcholinesterase,
butyrylcholinesterase, and carbonic anhydrase isoenzymes, J. Enzyme Inhib. Med.
Chem. 31 (2016) 1698–1702.
[28] A. Atmaca, S.T. Yıldırım, I. Çelik, C.T. Gulcin, M.Ç. Supuran, Intermolecular
amination of allylic and benzylic alcohols leads to effective inhibitions of acet-
ylcholinesterase enzyme and carbonic anhydrase I and II isoenzymes, J. Biochem.
Mol. Toxicol. 32 (2018) e22173.
[51] P. Taslimi, E. Koksal, A.C. Gören, E. Bursal, A. Aras, O. Kılıç, S. Alwasel, I. Gulcin,
Anti-Alzheimer, antidiabetic and antioxidant potential of Satureja cuneifolia and
analysis of ıts phenolic contents by LC-MS/MS, Arab. J. Chem. 13 (2020)
4528–4537.
[29] I. Gulcin, P. Taslimi, Sulfonamide inhibitors: A patent review 2013-present, Exp.
Opin. Therap. Pat. 28 (2018) 541–549.
[52] P. Taslimi, K. Turhan, F. Turkan, H.S. Karaman, Z. Turgut, I. Gulcin,
Cholinesterases, α-glycosidase, and carbonic anhydrase inhibition properties of
1H-pyrazolo[1,2-b]phthalazine-5,10-dione derivatives: Synthetic analogues for
the treatment of Alzheimer's disease and diabetes mellitus, Bioorg. Chem. 97
(2020) 103647.
[30] S. Burmaoglu, A.O. Yilmaz, P. Taslimi, O. Algul, D. Kılıç, I. Gulcin, Synthesis and
biological evaluation of phloroglucinol derivatives possessing α-glycosidase,
acetylcholinesterase, butyrylcholinesterase, carbonic anhydrase inhibitory ac-
tivity, Arch. Pharm. 351 (2018) e1700314.
[53] F. Turkan, M.N. Atalar, A. Aras, I. Gulcin, E. Bursal, ICP-MS and HPLC analyses,
7

İ. Gülçin, et al.
BioorganicChemistry103(2020)104171
enzyme inhibition and antioxidant potential of Achillea schischkinii Sosn, Bioorg.
Alzheimer's agents, Bioorg. Chem. 93 (2019) 103312.
Chem. 94 (2020) 103333.
[81] Y. Tao, Y.F. Zhang, Y.Y. Cheng, Y. Wang, Rapid screening and identification of
alpha-glucosidase inhibitors from mulberry leaves using enzyme- immobilized
magnetic beads coupled with HPLC/MS and NMR, Biomed. Chromatogr. 27
(2013) 148–155.
[54] A. Bytyqi-Damoni, A. Kestane, P. Taslimi, B. Tuzun, M. Zengin, H. Genç Bilgiçli,
I. Gulcin, Synthesis of novel β-amino carbonyl derivatives and their inhibition
effects on some metabolic enzymes, J. Mol. Struct. 1202 (2020) 127297.
[55] A. Biçer, R. Kaya, G. Yakali, M.S. Gültekin, G. Turgut Cin, I. Gulcin, Synthesis of
novel β-amino carbonyl derivatives and their inhibitioneffects on some metabolic
enzymes, J. Mol. Struct. 1204 (2020) 127453.
[82] Y. Demir, L. Durmaz, P. Taslimi, I. Gulcin, Anti-diabetic properties of dietary
phenolic compounds: ınhibition effects on α-amylase, aldose reductase and α-
glycosidase, Biotechnol. Appl. Biochem. 66 (2019) 781–786.
[56] A. Bytyqi-Damoni, A. Kestane, P. Taslimi, B. Tüzün, M. Zengin, H. Genç Bilgiçli,
I. Gulcin, Novel carvacrol based new oxypropanolamine derivatives: Design,
synthesis, characterization, biological evaluation, and molecular docking studies,
J. Mol. Struct. 1202 (2020) 127297.
[83] A. Aras, E. Bursal, F. Turkan, H. Tohma, O. Kılıç, I. Gulcin, E. Köksal,
Phytochemical content, antidiabetic, anticholinergic, and antioxidant activities of
endemic Lecokia cretica extracts, Chem. Biodiver. 16 (2019) e1900341.
[84] I. Gulcin, R. Kaya, A.C. Gören, H. Akıncıoğlu, M. Topal, Z. Bingöl, K. Çetin
Çakmak, S.B. Ozturk Sarikaya, L. Durmaz, S. Alwasel, Anticholinergic, antidiabetic
and antioxidant activities of Cinnamon (Cinnamomum verum) bark extracts:
Polyphenol contents analysis by LC-MS/MS, Int. J. Food Prop. 22 (2019)
1511–1526.
[57] I. Gulcin, A.C. Gören, P. Taslimi, B. Akyuz, B. Tüzün, Anticholinergic, antidiabetic
and antioxidant activities of Anatolian pennyroyal (Mentha pulegium)-Analysis of
its polyphenol contents by LC-MS/MS. Biocatal, Agric. Biotechnol. 23 (2020)
101441.
[58] A. Altay, H. Tohma, L. Durmaz, P. Taslimi, M. Korkmaz, I. Gulcin, E. Koksal,
Preliminary phytochemical analysis and evaluation of in vitro antioxidant, anti-
proliferative, antidiabetic and anticholinergics effects of endemic Gypsophila taxa
from Turkey, J. Food Biochem. 43 (2019) e12908.
[85] P. Taslimi, F. Turkan, A. Cetin, H. Burhan, M. Karaman, İ. Bildirici, I. Gulcin,
F. Şen, Pyrazole[3,4-d]pyridazine derivatives: Molecular docking and explore of
acetylcholinesterase and carbonic anhydrase enzymes inhibitors as antic-
holinergics potentials, Bioorg. Chem. 92 (2019) 103213.
[59] K. Cetin Cakmak, I. Gulcin, Anticholinergic and antioxidant activities of usnic
acid-An activity-structure insight, Toxicol. Rep. 6 (2019) 1273–1280.
[60] P. Taslimi, I. Gulcin, B. Ozgeris, S. Goksu, F. Tumer, S.H. Alwasel, C.T. Supuran,
The human carbonic anhydrase isoenzymes I and II (hCA I and II) inhibition effects
of trimethoxyindane derivatives, J. Enzyme Inhib. Med. Chem. 31 (2016)
152–157.
[86] C. Caglayan, P. Taslimi, C. Türk, F.M. Kandemir, Y. Demir, I. Gulcin, Purification
and characterization of the carbonic anhydrase enzyme from horse mackerel
(Trachurus trachurus) muscle and the impact of some metal ions and pesticides on
enzyme activity, Comp. Biochem. Physiol. C 226 (2019) 108605.
[87] M. Kucuk, I. Gulcin, Purification and characterization of carbonic anhydrase en-
zyme from black sea trout (Salmo trutta Labrax Coruhensis) kidney and inhibition
effects of some metal ions on the enzyme activity, Environ. Toxicol. Pharmacol. 44
(2016) 134–139.
[61] A. Yıldırım, U. Atmaca, A. Keskin, M. Topal, M. Çelik, I. Gulcin, C.T. Supuran, N-
Acylsulfonamides strongly inhibit human carbonic anhydrase isoenzymes I and II,
Bioorg. Med. Chem. 23 (2015) 2598–2605.
[88] (a) H. Genc Bilgicli, D. Ergon, P. Taslimi, B. Tuzun, I. Akyazı Kuru, M. Zengin,
I. Gulcin, Novel propanolamine derivatives attached to 2-metoxifenol moiety:
Synthesis, characterization, biological properties, and molecular docking studies,
Bioorg. Chem. 101 (2020) 103969;
[62] A. Atasever, H. Özdemir, I. Gulcin, Ö.İ. Küfrevioğlu, One-step purification of
lactoperoxidase from bovine milk by affinity chromatography, Food Chem. 136
(2013) 864–870.
[63] I. Gulcin, S. Beydemir, Phenolic compounds as antioxidants: Carbonic anhydrase
isoenzymes inhibitors, Mini Rev. Med. Chem. 13 (2013) 408–430.
[64] A. Akıncıoğlu, Y. Akbaba, H. Göçer, S. Göksu, I. Gulcin, C.T. Supuran, Novel
sulfamides as potential carbonic anhydrase isoenzymes inhibitors, Bioorg. Chem.
21 (2013) 1379–1385.
(b) N. Lolak, S. Akocak, C. Turkes, P. Taslimi, M. Isik, Ş. Beydemir, I. Gulcin,
M. Durgun, Synthesis, characterization, inhibition effects, and molecular docking
studies as acetylcholinesterase, α-glycosidase, and carbonic anhydrase inhibitors
of novel benzenesulfonamides incorporating 1,3,5-triazine structural motifs,
Bioorg. Chem. 100 (2020) 103897;
[65] K. Aksu, M. Nar, M. Tanç, D. Vullo, I. Gulcin, S. Göksu, F. Tümer, C.T. Supuran,
Synthesis and carbonic anhydrase inhibitory properties of sulfamides structurally
related to dopamine, Bioorg. Med. Chem. 21 (2013) 2925–2931.
[66] Y. Akbaba, A. Akıncıoğlu, H. Göçer, S. Göksu, I. Gulcin, C.T. Supuran, Carbonic
anhydrase inhibitory properties of novel sulfonamide derivatives of aminoindanes
and aminotetralins, J. Enzyme Inhib. Med. Chem. 29 (2014) 35–42.
[67] M. Topal, I. Gulcin, Rosmarinic acid: a potent carbonic anhydrase isoenzymes
inhibitor, Turk. J. Chem. 38 (2014) 894–902.
(c) T. Artunç, A. Menzek, P. Taslimi, I. Gulcin, C. Kazaz, E. Sahin, Synthesis and
antioxidant activities of phenol derivatives from 1,6-bis(dimethoxyphenyl)hexane-
1,6-dione, Bioorg. Chem. 100 (2020) 103884.
[89] B.A. Trofimov, A.I. Mikhaleva, E.Y. Schmidt, L.N. Sobenina, Chemistry of Pyrroles,
CRC Press Inc: Boca-Raton (2014).
[90] S.F. Malysheva, V.A. Kuimov, A.B. Trofimov, N.A. Belogorlova, Y.I. Litvintsev,
A.M. Belogolova, N.K. Gusarova, B.A. Trofimov, 2-Halopyridines in a triple reac-
tion in the Pn/KOH/DMSO system to form tri(2-pyridyl)phosphine: experimental
and quantum-chemical dissimilarities, Mendeleev Commun. 28 (2018) 472–474.
[91] A.N. Kharat, A. Bakhoda, T. Hajiashrafi, A. Abbasi, Synthesis, Characterization,
and crystal structures of tris(2-pyridyl)phosphine sulfide and selenide,
Phosphorus, Sulfur and Silicon 185 (2010) 2341–2347.
[68] B. Arabaci, I. Gulcin, S. Alwasel, Capsaicin: A potent inhibitor of carbonic anhy-
drase isoenzymes, Molecules 19 (2014) 10103–10114.
[69] H. Göçer, A. Akıncıoğlu, S. Göksu, I. Gulcin, C.T. Supuran, Carbonic anhydrase and
acetylcholine esterase inhibitory effects of carbamates and sulfamoylcarbamates,
J. Enzyme Inhib. Med. Chem. 30 (2015) 316–320.
[92] M. Sentürk, I. Gulcin, A. Dastan, Ö.İ. Küfrevioğlu, C.T. Supuran, Carbonic anhy-
drase inhibitors. Inhibition of human erythrocyte isozymes I and II with a series of
antioxidant phenols, Bioorg Med Chem 17 (2009) 3207–3211.
[70] M. Boztaş, Y. Çetinkaya, M. Topal, I. Gulcin, A. Menzek, E. Şahin, M. Tanc,
C.T. Supuran, Synthesis and carbonic anhydrase isoenzymes I, II, IX, and XII in-
hibitory effects of dimethoxy-bromophenol derivatives incorporating cyclopro-
pane moieties, J. Med. Chem. 58 (2015) 640–650.
[93] C.T. Supuran, A. Scozzafava, A. Casini, Carbonic anhydrase inhibitors, Med. Res.
Rev. 23 (2003) 146–189.
[71] M.M. Bradford, A rapid and sensitive method for the quantitation of microgram
quantities of protein utilizing the principle of protein-dye binding, Anal. Biochem.
72 (1976) 248–251.
[94] T.A. Coban, S. Beydemir, I. Gulcin, D. Ekinci, A. Innocenti, D. Vullo, C.T. Supuran,
Sildenafil is a strong activator of mammalian carbonic anhydrase isoforms I-XIV,
Bioorg. Med. Chem. 17 (2009) 5791–5795.
[72] A. Yıldırım, U. Atmaca, A. Keskin, M. Topal, M. Çelik, I. Gulcin, C.T. Supuran, N-
Acylsulfonamides strongly inhibit human carbonic anhydrase isoenzymes I and II,
Bioorg. Med. Chem. 23 (2015) 2598–2605.
[95] S.B. Oztürk Sarikaya, I. Gulcin, C.T. Supuran, Carbonic anhydrase inhibitors.
Inhibition of human erythrocyte isozymes I and II with a series of phenolic acids,
Chem. Biol. Drug Des. 75 (2010) 515–520.
[73] D.K. Laemmli, Cleavage of structural proteins during the assembly of the head of
bacteriophage T4, Nature 227 (1970) 680–683.
[96] M. Şentürk, I. Gulcin, Ş. Beydemir, Ö.İ. Küfrevioğlu, C.T. Supuran, In vitro in-
hibition of human carbonic anhydrase I and II isozymes with natural phenolic
compounds, Chem. Biol. Drug. Des. 77 (2011) 494–499.
[74] A. Scozzafava, M. Passaponti, C.T. Supuran, I. Gulcin, Carbonic anhydrase in-
hibitors: Guaiacol and catechol derivatives effectively inhibit certain human car-
bonic anhydrase isoenzymes (hCA I, II, IX, and XII), J. Enzyme Inhib. Med. Chem.
30 (2015) 586–591.
[97] A. Innocenti, S.B. Öztürk Sarikaya, I. Gulcin, C.T. Supuran, Carbonic anhydrase
inhibitors. Inhibition of mammalian isoforms I-XIV with a series of natural product
polyphenols and phenolic acids, Bioorg. Med. Chem. 18 (2010) 2159–2164.
[98] A. Innocenti, I. Gulcin, A. Scozzafava, C.T. Supuran, Carbonic anhydrase in-
hibitors. Antioxidant polyphenol natural products effectively inhibit mammalian
isoforms I-XV, Bioorg. Med. Chem. Let. 20 (2010) 5050–5053.
[75] H. Gocer, A. Aslan, I. Gulcin, C.T. Supuran, Spirobisnaphthalenes effectively in-
hibit carbonic anhydrase, J. Enzyme Inhib. Med. Chem. 31 (2016) 503–507.
[76] G.L. Ellman, K.D. Courtney, V. Andres, R.M. Featherston, A new and rapid col-
orimetric determination of acetylcholinesterase activity, Biochem. Pharmacol. 7
(1961) 88–95.
[99] S.B. Oztürk Sarikaya, F. Topal, M. Şentürk, İ. Gülçin, C.T. Supuran, In vitro in-
hibition of α-carbonic anhydrase isozymes by some phenolic compounds, Bioorg.
Med. Chem. Let. 21 (2011) 4259–4262.
[77] S. Karakaya, Z. Bingol, M. Koca, S. Dagoglu, N.M. Pınar, B. Demirci, I. Gulcin,
M. Brestic, O. Sytar, Identification of non-alkaloid natural compounds of Angelica
purpurascens (Avé-Lall.) Gilli. (Apiaceae) with acetylcholinesterase and cholines-
terase inhibition potential, Saudi Pharm. J. 28 (2020) 1–14.
[100] M. Nar, Y. Çetinkaya, I. Gulcin, A. Menzek, (3,4-Dihydroxyphenyl) (2,3,4-trihy-
droxyphenyl)methanone and its derivatives as carbonic anhydrase isoenzymes
inhibitors, J. Enzyme Inhib. Med. Chem. 28 (2013) 402–406.
[78] H. Genç Bilgiçli, P. Taslimi, B. Akyuz, B. Tüzün, I. Gulcin, Synthesis, character-
ization, biological evaluation, and molecular docking studies of some piperonyl
based 4-thiazolidinone derivatives, Arch. Pharm. 353 (2020) 1900304.
[79] G. Mamedova, A. Mahmudova, S. Mamedov, Y. Erden, P. Taslimi, B. Tüzün, R. Taş,
V. Farzaliyev, A. Sujayev, S.H. Alwasel, I. Gulcin, Novel tribenzylamino-
benzolsulphonylimine based on their pyrazine and pyridazines: synthesis, char-
acterization, antidiabetic, anticancer, anticholinergics, and molecular docking
studies, Bioorg. Chem. 93 (2019) 103313.
[101] P. Taslimi, I. Gulcin, N. Öztaşkın, Y. Çetinkaya, S. Göksu, S.H. Alwasel,
C.T. Supuran, The effects of some bromophenol derivatives on human carbonic
anhydrase isoenzymes, J. Enzyme Inhib. Med. Chem. 31 (2016) 603–607.
[102] H. Göksu, M. Topal, A. Keskin, M.S. Gültekin, M. Çelik, I. Gulcin, M. Tanc,
C.T. Supuran, 9,10-Dibromo-N-aryl-9,10-dihydro-9,10-[3,4]epipyrroloan-
thracene-12,14-diones; synthesis and investigation of their effects on carbonic
anhydrase isozymes I, II, IX, and XII, Arch. Pharm. 349 (2016) 466–474.
[103] C.T. Supuran, A. Di Fiore, G. De Simone, Carbonic anhydrase inhibitors as emer-
ging drugs for the treatment of obesity, Exp. Opin. Emerg. Drug. 13 (2008)
383–392.
[80] N.A.E. El-Sayed, A.E.S. Farag, M.A.F. Ezzat, H. Akincioglu, I. Gulcin, S.M. Abou-
Seri, Design, synthesis, in vitro and in vivo evaluation of novel pyrrolizine-based
compounds with potential activity as cholinesterase inhibitors and anti-
[104] H.T. Abdel-Mohsen, A.E. El Kerdawy, M.A. Omar, E. Berrino, A.S. Abdelsamie,
8

İ. Gülçin, et al.
H.I. El Diwani, C.T. Supuran, New thiopyrimidine-benzenesulfonamide conjugates
BioorganicChemistry103(2020)104171
mellitus and Alzheimer's disease: shared pathology and treatment? Br. J. Clin.
as selective carbonic anhydrase II inhibitors: synthesis, in vitro biological eva-
luation, and molecular docking studies, Bioorg. Med. Chem. 28 (2020) 115329.
[105] H. Genç, R. Kalin, Z. Köksal, N. Sadeghian, U.M. Kocyigit, M. Zengin, I. Gulcin,
H. Ozdemir, Discovery of potent carbonic anhydrase and acetylcholinesterase in-
hibitors: 2-aminoindan β-lactam derivatives, Int. J. Mol. Sci. 17 (2016) 1736.
[106] O.M. Khalil, A.M. Kamal, S. Bua, H. El Sayed Teba, Y.M. Nissan, C.T. Supuran,
Pyrrolo and pyrrolopyrimidine sulfonamides act as cytotoxic agents in hypoxia via
inhibition of transmembrane carbonic anhydrases, Eur. J. Med. Chem. 188 (2020)
112021.
Pharmacol. 71 (2011) 365–376;
b S.M. de la Monte, J.R. Wands, Alzheimer's disease is type 3 diabetes-Evidence
reviewed, J. Diabet. Sci. Technol. 2 (2008) 1101–1113.
[108] a J.N. Fawver, Y. Ghiwot, C. Koola, W. Carrera, J. Rodriguez-Rivera,
C. Hernandez, K.T. Dineley, Y. Kong, J. Li, J. Jhamandas, G. Perry, I.V.J. Murray,
Islet amyloid polypeptide (IAPP): A second amyloid in Alzheimer's disease, Curr.
Alzheimer Res. 11 (2014) 928–940;
b B.B. Bendlin, Antidiabetic therapies and Alzheimer disease, Dialogues Clin.
Neurosci. 21 (2019) 83–91.
[107] a K. Akter, E.A. Lanza, S.A. Martin, N. Myronyuk, M. Rua, R.B. Raffa, Diabetes
9