Synthesis, characterization and biological evaluation of N-substituted triazinane-2-thiones and theoretical–experimental mechanism of condensation reaction

Article abstract of DOI:10.1002/aoc.5329

The synthesis of triazinthions and their reactions with some nucleophilic reagents have been investigated during this scientific study. Thus, thiourea with a single component has been synthesized as a result of concomitant reactions of aldehyde and amines

Full text of DOI:10.1002/aoc.5329

Received: 5 September 2019
DOI: 10.1002/aoc.5329
Revised: 8 October 2019
Accepted: 12 October 2019
F U L L P A P E R
Synthesis, characterization and biological evaluation of N-
substituted triazinane-2-thiones and theoretical–
experimental mechanism of condensation reaction
Afsun Sujayev1 | Parham Taslimi2
Lala Aliyeva1 | Vagif Farzaliyev1 | Ilhami Gulçin
| Ruya Kaya3,4 | Bahruz Safarov1
|
3
_
1Laboratory of Theoretical Bases of
The synthesis of triazinthions and their reactions with some nucleophilic
reagents have been investigated during this scientific study. Thus, thiourea
with a single component has been synthesized as a result of concomitant reac-
tions of aldehyde and amines trials. The structure of the synthesized com-
Synthesis and Action Mechanism of
Additives, Institute of Chemistry of
Additives, Azerbaijan National Academy
of Sciences, 1029, Baku, Azerbaijan
2Department of Biotechnology, Faculty of
Science, Bartin University, 74100, Bartin,
Turkey
3Department of Chemistry, Faculty of
Sciences, Ataturk University, 25240,
Erzurum, Turkey
4Central Research and Application
Laboratory, Agri Ibrahim Cecen
University, 04100, Agri, Turkey
pounds was confirmed by H, 13C NMR spectroscopy methods. The inhibitory
1
effects of novel N-substituted triazinane-2-thione derivatives on acetylcholines-
terase (AChE) activity were performed according to the spectrophotometric
method of Ellman et al. These novel N-substituted triazinane-2-thiones deriva-
tives were effective inhibitors of the α-glycosidase, cytosolic carbonic
anhydrase I and II isoforms (hCA I and II), and Acetylcholinesterase (AChE)
enzymes with Ki values in the range of 1.01 0.28 to 2.12 0.37 nM for α-gly-
Correspondence
cosidase, 13.44
4.39 to 74.98
6.25 nM for hCA I, 10.41
4.8 to
Parham Taslimi, Faculty of Science,
Department of Biotechnology, Bartin
University, 74100-.Bartin, Turkey.
Email: parham_taslimi_un@yahoo.com;
ptaslimi@bartin.edu.tr
72.6 17.66 nM for hCA II, 36.82 9.95 to 108.48 1.17 nM for AChE, and
624.62 100.34 to 1124.16 205.14 nM for α-glycosidase, respectively.
K E Y W O R D S
activation energy, enzyme inhibition, metabolic enzymes, N-substituted triazines
1 | INTRODUCTION
pharmaceutically important natural and synthetic
materials, showing powerful ectoparasiticidal action,
Among the heterocyclic compounds, triazine deriva-
tives are attracting interest in medicinal and industrial
chemistry and they are also being developed as a pre-
cursor for the synthesis of nitrogen-containing organic
compounds. The triazine and hexahydrotriazine
(triazinane) nuclei constitute well-known compounds
that have been used as an anticonvulsant, anti-
coccidial, antiplasmodial, antimalarial and herbicidal
agents. In addition, some of these compounds can
serve as hydrogen sulfide scavengers, chiral discrimina-
tors and low-toxicity drug deliverers. Furthermore, het-
erocyclic compounds containing a thiourea or urea
potent antidiabetic properties and anti-HIV activity. A
method of synthesis for 4,6-diphenyl-1,3,5-triazinane-
2-thione was developed by three-component condensa-
tion of thiourea, benzaldehyde and ammonia.[1] Taking
into consideration the above, we have continued to
investigate the synthesis of new triazithione derivatives
and their biological activity through triple-component
condensation.
Carbonic anhydrase (CA, E.C.4.2.1.1) enzymes are
very efficient enzymes, which catalyze the hydration of
carbon
dioxide
to
produce
a
proton
and
bicarbonate.[2–4] This mechanism is common for most
organisms, and as an outcome of the evolution of
structural unit have
a
special place among
Appl Organometal Chem. 2019;e5329.
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© 2019 John Wiley & Sons, Ltd.
1 of 16
https://doi.org/10.1002/aoc.5329

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SUJAYEV ET AL.
seven genetically varying families, this enzyme has also
evolved to[5,6] α-CA enzymes. Sixteen different α-CA
isozymes have been characterized and isolated in verte-
brate and mammal cells so far, varying in cellular
localization (mitochondria, cytosol, membrane) and tis-
sue distribution.[7,8] Some isozymes have also been
studied in detail by means of X-ray crystallography,
recording images of the enzyme active site, where a
zinc ion plays an axial role in the catalytic activity.[9]
Indeed, other structural specifications of the enzyme
active site have been highlighted, such as the presence
of two diverse areas, one with hydrophilic amino acids
and the other one lined with hydrophobic residues,
offering the chance to modulate ligand design in dispa-
rate ways.[10,11] CA inhibitor compounds have been
clinically used for almost 60 years as antiglaucoma
and diuretics drugs.[12]
arterial hypertension. Recent scientific research shows
that various substances isolated from plant tissues have a
very important hypoglycemic efficacy.[29]
In this study, we have performed a facile synthesis of
the well-defined theoretical–experimental mechanism of
the condensation reaction of novel N-substituted
triazinane-2-thiones derivatives and also investigated
their inhibition potential of cytosolic carbonic anhydrase
I and II isoforms (hCA I and II), to discover the most
favorable and potent AChE and α-glycosidase inhibition
properties of these compounds.
2 | MATERIALS AND METHODS
2.1 | General chemistry
Acetylcholinesterase (AChE) is a significant enzyme
that catalyzes the breakdown of acetylcholine (ACh)
and some other choline ester molecules that function
as neurotransmitter molecules, which are recorded as a
drugs for Alzheimer's disease (AD).[13–16] The choliner-
gic hypothesis process was first proposed to describe
AD and was based on the finding that the synaptic
depression is hindered owing to the prevention of the
ACh compound hydrolysis in the cholinergic neuron
cells.[17,18] The inhibition action of AChE enzyme
results in the blockage of ACh hydrolysis. Indeed, the
design of inhibitor compounds or/and modulators for
the AChE enzyme has been of major interest since it
is customarily one of the best ways to prevent AD,
resulting in three commercial drug compounds
being approved by the US Food and Drug Administra-
tion, comprising rivastigmine, galantamine and
donepezil.[19,20] Many potential inhibitors of AChE
Solvents were distilled under anhydrous conditions. All
reagents were purchased and used without further purifi-
cation. Glassware was dried prior to use. Compounds
were purified by dry flash chromatography using silica
60 < 0063 mm and water pump vacuum or by flash-
chromatography using silica 60 Å 230–400 mesh as sta-
tionary phases. Thin-layer chromatography (TLC) plates
(silica gel 60 F254) were visualized either at a UV lamp or
in iodine. Melting points are uncorrected. Elemental
analysis was performed on the Carlo Erba 1108 analyzer.
2.2 | NMR experiments
NMR experiments were performed on a Bruker FT NMR
1
spectrometer Avance 300 (300 MHz for H and 75 MHz
for 13C) with BVT 3200 variable-temperature unit in
5 mm sample tubes using Bruker Standard software. The
1H and 13C chemical shifts were referenced to internal
tetramethylsilane. NMR-grade DMSO-d6 (99.7%, con-
taining 0.3% H2O) was used for the synthesized
compounds.
enzyme are under investigation, such as
tacrine
derivatives and Zijuan tea. Some of these derivatives
are under clinical evaluation such as tacrine,
huperzine A and ganstigmine.[21–24]
Recently, diverse hypoglycemic drug compounds have
been utilized to lower the level of blood glucose in dia-
betic patients (like biguanides and sulfonylurea as well as
other novel classes).[25,26] Between these classes are the
inhibitor compounds of α-glycosidase – an enzyme which
is accountable for the hydrolysis of carbohydrate mole-
cules to facilitate the transmission of glucose molecules
towards the blood compartment.[27] Presently, two kinds
of drug compounds are utilized to inhibit the enzymatic
activities of this enzyme in the intestine: the miglitol and
the acarbose, which decrease postprandial hyperglyce-
mia.[28] The inhibitor compounds of this enzyme not only
have a beneficial effect on the postprandial glycaemia but
also have a beneficial action against viral agents and
2.3 | Kinetic approach
To illustrate the efficient catalytic route, the energetic
span model (δG, here in terms of the Gibbs energy) was
applied to demonstrate feasibility of the blue, orange and
grey routes (see Figure 1). According to the energetic
span model, the lowest energetic intermediate and the
highest energetic transition state are determined for each
route. The δG value was found to be the similar for blue
(trifluoroacetic acid, TFA) and grey (acetic acid, AA)
routes: 0.1 kcal/mol difference was observed. The δG
value of the orange route (trichloroacetic acid, TCA) was

SUJAYEV ET AL.
3 of 16
FIGURE 1 Reaction profile
(kcal/mol) for the OTC triazinane-
2-thione formation according to density
functional theory (DFT) calculations.
Blue, orange and grey colors correspond
to the trifluoroacetic acid (TFA),
trichloroacetic acid (TCA) and acetic
acid (AA) catalyzed routes
2 kcal/mol less than the grey route. The TOF (turnover
frequency) values were calculated for each route via
using the Eyring equation (Equation (1)):
catalyst was inserted. The resulting solution was stirred
for 12–24 h at reflux. When the reaction mixture was
cooled to room temperature, a white solid precipitated.
The precipitates were filtered and recrystallized from
Ethyl acetate (EtOAc) after dichloromethane washing
and gave the product (4,6-disubstituted 1,3,5-triazinane-
2-thiones, 1a–g) in 65–85% yield.
kBT −δG
RT
TOF =
e
ð1Þ
h
where kB stands for Boltzmann's constant, h is Plank's
constant, R is the universal gas constant and T is
temperature in Kelvin. The following calculations
[equations (2) and (3)] show us that there is no a
dramatic difference in the catalytic efficiencies of TFA,
TCA, and AA on the cyclocondenstation reaction:
2.4.2 | 5-(1,5-Diaminopentan-3-yl)-4,6-
bis(2-hydroxyphenyl)-1,3,5-triazinane-2-
thione (1a)
1H NMR (300 MHz, DMSO-d6, δ): d (mixture of two
diastereoisomers) = 1.92 (t, J = 12.0 Hz, 1 H, NH), 2.0 m
(2H, NH2), 2.48, 2.65, 2.77, 2.81 (d, J = 7.75 Hz, 2H,
CH2), 3.03 (1H, NH), 5.04 (d, J = 12 Hz, 2H, 2CH–tri-
azin), 6.61–7.04 (m, 20 H, CH–Ar), 7.31 (s, 1H, NH),
9.83 m (1H, PhOH).
TOFacetic acid
= 1:17
ð2Þ
ð3Þ
TOFtrifluoroacetic acid
TOFtrichloroacetic acid
TOFacetic acid
= 24:91
13C NMR (75 MHz, DMSO-d6): d (single diastereo-
isomer, trans) = 36.5 (-CH2), 57.7 (-CH2), 70.3 (-CH–
triazinane), 115.7, 119.9, 121.2, 128.5, 129.3 (-CH–Ar),
155 (-Ph–OH), 177.7 (-C=S).
2.4 | Synthesis of new 4,6-disubstituted
1,3,5-Triazinane-2-thiones (1a–g)
2.4.1 | General procedure
2.4.3 | 5-Methyl-4,6-bis(p-tolyl)-1,3,5-
triazinane-2-thione (1b)
The aldehydes (salicylic aldehyde, 4-methylbenzal-
dehyde, anisylaldehyde and benzaldehyde, 0.02–0.2 mol)
were added to NH4OH (33,5%, 15 ml) or amines
(diethylenetryamine, methylamine and ethylamine,
0.01–0.05 mol) and the solution was stirred for 2–3 h at
reflux. During this time, a white precipitate formed. The
precipitate was removed by filtration and dried. The solid
was dissolved in isopropyl alcohol (20 ml), thiourea
(0.05–0.15 mol) was added to the mixture. Upon being
dissolved in the stirring rod within 30 min the TFA
1H NMR (300 MHz, DMSO-d6, δ): d (mixture of two
diastereoisomers) = 2.0 (t, J = 11.5 Hz, 1H, NH), 2.19 (s,
6H, 2CH3–Ar), 2.27 (s, 3H, CH3–N), 5.04 (d, J = 12 Hz,
2H, 2CH–triazin), 6.94 (m, 8H, CH–Ar), 7.31 (br,
1H, NH).
13C NMR (75 MHz, DMSO-d6): d (single diaste-
reoisomer, trans) = 24.3 (CH3–Ar-), 32.7 (-N–CH3),
79 (-CH–triazinane), 128.7, 135.3, 136.9 (-CH–Ar),
177.4 (C=S).

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SUJAYEV ET AL.
2.4.4 | 5-Ethyl-4,6-bis(2-hydroxyphenyl)-
1,3,5-triazinane-2-thione (1c)
13C NMR (75 MHz, DMSO-d6): d (single diastereo-
isomer, trans) = 32.7 (CH3–N-), 55.7 (CH3O-), 79 (-CH–
triazinane), 114, 129.8, 130.6, 159.1 (-CH–Ar),
177.2 (C=S).
1H NMR (300 MHz, DMSO-d6, δ): d (single diastereo-
isomer, trans) = 1.00 (s, 3H, CH3), 2.02 (t, J = 8.5 Hz, 1H,
NH), 2.40 (d, J = 8.5 Hz, 2H, CH2–N), 5.04 (d, J = 8.5 Hz,
2H, 2CH), 6.61, 6.70, 6.89, 7.04 (m, 1H, CH–Ar), 9.83 m
(1H, OH).
2.4.8 | 5-Ethyl-4,6-bis(4-methoxyphenyl)-
1,3,5-triazinane-2-thione (1g)
13C NMR (75 MHz, DMSO-d6): d (single diastereo-
isomer, trans) = 13.3 (-CH3), 38.8 (-CH2–N-), 70.3 (-CH–
triazinane), 115.6, 119, 121.1, 128.7, 130.2 (-CH–Ar),
155.9 (-Ph–OH), 177.6 (C=S).
1H NMR (300 MHz, DMSO-d6, δ): d (mixture of two
diastereoisomers)
=
1.01 (t, 3H, CH3), 2.02 (t,
J = 11.5 Hz, 1H, NH), 2.40 (s, 2H, CH2–N-), 3.72 (s, 6H,
2OCH3), 5.02 (d, 2H, 2CH), 6.65 (d, J = 11.5 Hz, 4H,
2CH), 7.39 (m, 4H, 2CH–Ar).
2.4.5 | 4,6-Diphenyl-1,3,5-triazinane-2-
thione (1d)
13C NMR (75 MHz, DMSO-d6): d (single diastereo-
isomer, trans) = 13.3 (-CH3), 38.8 (-CH2–N-), 55.8
(CH3O-), 76.5 (-CH–triazinane), 114, 129.8, 130.6, 159.1 (-
CH–Ar), 178 (C=S).
1H NMR (DMSO-d6, 300 MHz): d (mixture of two dia-
stereoisomers) = 2.01 (t, J = 11.75 Hz, 1 H, NH), 2.55 (t,
J = 8.75 Hz, 1 H, NH), 5.21 (d, J = 8.75 Hz, 2 H), 5.54 (d,
J = 12 Hz, 2 H), 6.91 (s, 2 H, NH), 7.21 (s, 2 H, NH),
7.30–7.50 (m, 20 H).
2.4.9 | 1,3-bis(2-Hydroxybutyl)-5-methyl-
4,6-bis(p-tolyl)-1,3,5-triazinane-2-thione (2a)
13C NMR (DMSO-d6, 75 MHz): d (mixture of two
diastereoisomers) = 65.7 (-CH–triazinane), 69.6 (-CH–
triazinane), 126.4, 126.6, 128.9, 129.0, 129.2, 129.7, 137.1,
138.3 (-CH–Ar), 178.4 (C=S).
5-Methyl-4,6-bis(p-tolyl)-1,3,5-triazinane-2-thion
(1b)
(3.11 g, 0.01 mol) was dissolved in a 2:1 ratio of
acetylacetone and ethyl alcohol (12:5 ml) and
1,2-epoxobutane (2.03 ml, 0.02 mol) was added drop by
drop. After being dissolved in the stirrer for 30 min,
0.02 g AlCl3 catalyst was added and mixed by heating at
60–65ꢀC. After determining the full completion of reac-
tion, the solution was evaporated and cleaned in ethyl
alcohol solution.
The compound (1d) was also synthesized according to
1
above procedure and their H- and 13C-NMR data are in
agreement with data given in the literature.[1]
2.4.6 | 5-Ethyl-4,6-bis(p-tolyl)-1,3,5-
triazinane-2-thione (1e)
Analogously,
4,6-bis(p-tolyl)-1,3,5-triazinane-2-thione (2b) was synthe-
sized by the reaction of triazinanes with
1,2-epoxypropane.
1,3-bis(2-hydroxypropyl)-5-methyl-
1H NMR (300 MHz, DMSO-d6, δ): d (mixture of two
diastereoisomers)
=
1.10 (t, 3H, CH3), 1.94 (t,
J = 12.0 Hz, 1H, NH), 2.19 (s 6H, 2CH3–Ar), 2.40 (d,
J = 6.0 Hz, 2H, CH2–N-), 5.04 m (d, J = 12 Hz, 2H, 2CH),
6.9 (m 8H, CH–Ar).
(2a) compound 1H NMR (300 MHz, DMSO-d6, δ): d
(mixture of two diastereoisomers) = 0.96 (t, 3H, CH3), 1.48
(d, J = 8.5 Hz, 2H, CH2), 1.78 (t, J = 12.25 Hz, 1 H, NH),
2.27 (s, 3H, CH3), 2.28 (t, J = 8.5 Hz, 1H, NH), 3.45 (d, 1H,
CH), 3.47, 3.72, (d, J = 12.25 Hz, 2H, CH2), 4.81 (d, 1H,
OH), 5.04 (d, 2H, 2CH–triazine), 6.9 (m, 8H, CH–Ar).
13C NMR (75 MHz, DMSO-d6): d (mixture of two
diastereoisomers) = 7.6 (CH3-Alk), 24.3 (CH3–Ar), 28.6 (-
CH2-), 33 (CH3–N-), 56.5 (-CH2-), 71.2 [-CH (OH)], 79.3
(-CH–triazinane), 81.7 (-CH–triazinane), 128.7, 135.3,
136.9 (-CH–Ar), 177.3 (C=S).
13C NMR (75 MHz, DMSO-d6): d (single diastereo-
isomer, trans) = 15.5 (-CH3), 22.4 (CH3–Ar-), 24.3 (CH3–
Ar-), 39.5 (-CH2–N-), 49 (-CH–triazinane), 74.8 (-CH–
triazinane), 116.1, 127.8, 128.8, 129.3, 130.4, 136.2, 136.6,
138.3 (-CH–Ar), 182 (C=S).
2.4.7 | 5-Methyl-4,6-bis(4-
methoxyphenyl)-1,3,5-triazinane-2-
thione (1f)
1
(2b) compound - H NMR (300 MHz, DMSO-d6,
1H NMR (300 MHz, DMSO-d6, δ): d (mixture of two
diastereoisomers) = 2.27 (s, 3H, CH3–N-), 2.30 (t,
J = 12 Hz, 1 H, NH), 2.72 (t, J = 8.25 Hz, 1 H, NH), 3.72
(s, 6H, 2OCH3), 5.02 (d, J = 8.25 Hz, 2H, 2CH), 6.65 (m,
4H, 2CH–Ar), 7.39 (m, 4H, 2CH–Ar).
δ): d (mixture of two diastereoisomers) = 1.21 (t, 3H,
CH3), 2.19 (s, 3 H, CH3), 2.02 (t, J = 11.5 Hz, 1H, NH),
2.27 s (3H, CH3N), 3.47 3.72, (d, J = 11.5 Hz, 2H, CH2),
3.63 [d, 1H, CH (OH)], 4.81 (d, 1H, OH), 5.01 (s, 1H,
CH), 6.9 (m, 8H, CH–Ar).

SUJAYEV ET AL.
5 of 16
13C NMR (75 MHz, DMSO-d6): d (single diastereo-
CH2Cl), 53.9 (-CH2–N-), 74.1 [-CH (OH)], 79.3 (-CH–
triazinane), 81.7 (-CH–triazinane), 128.7, 135.9, 136.9 (-
CH–Ar), 177.3 (C=S).
isomer, trans) = 23 (-CH3-Alk), 24.3 (CH3–Ar), 33 (CH3–
N-), 59 (-CH2-), 66.2 [-CH (OH)], 79.3 (-CH–triazinane),
81.7 (-CH–triazinane), 128.7, 135.3, 136.9 (-CH–Ar),
177.3 (C=S).
2.5 | Biochemical studies
2.4.10 | 1,3-bis(4-Hydroxybutan-2-yl)-5-
methyl-4,6-bis(p-tolyl)-1,3,5-triazinane-2-
thione (2c)
2.5.1 | hCA Isoenzyme purification and
inhibition studies
For investigating of inhibitory effects of novel N-
substituted triazinane-2-thiones derivatives (1a–g and
2a–d) on hCA isoforms, both hCA isoforms from human
erythrocytes were purified via a simple single-step
method using Sepharose-4B-L-Tyrosine-sulphanilamide
affinity gel chromatography.[30,31] For this purpose, the
human erythrocyte samples were centrifuged at
13,000 rpm for 25 min.[32] Then, the solution was filtered
to remove the precipitate. Both hCA isoenzymes were
isolated from the serum, the pH of which was adjusted to
8.7 by adding solid Tris.[33] Affinity column was equili-
5-Methyl-4,6-di(p-tolyl)-1,3,5-triazinane-2-thion
(1b) (0.01 mol) was dissolved in 5–10 ml acetyl acetone
and ethyl alcohol, which were the reactive chemicals, in
a 2:1 molar ratio, and added to the solution slowly.
4-Chlor-butanol-1 was added and mixed for 10–15 min.
After that, triethylamine catalyst was added and mixed.
The solution was mixed at 70–78ꢀC temperature for
1–3 h. After the completion of the reaction, the mixture
was cooled, purified by means of re-crystallization with
ethanol and dried.
1H NMR (300 MHz, DMSO-d6, δ): d (single diaste-
reoisomer, trans) = 1.10 (t, 3H, CH3), 1.56 (d, 2H, CH2),
2.19 s (6H, 2CH3–Ar), 2.27 (s, 3H, CH3N), 2.29 (t,
J = 12.25 Hz, 1 H, NH), 2.79 (d, J = 12.25 Hz, 2H, CH2),
3.53 (d,1H, CH), 4.78 m (1H, OH), 6.94 m (8H, CH–Ar).
13C NMR (75 MHz, DMSO-d6): d (single diastereo-
isomer, trans) = 19 (-CH3-Alk), 24.3 (CH3–Ar), 33 (CH3–
N-), 38.8 (-CH2-), 50.8 (-CH–CH3), 58.3 (-CH2–OH), 78.9
(-CH–triazinane), 79.3 (-CH–triazinane), 128.7, 135.9,
136.9 (-CH–Ar), 176.9 (C=S).
brated by buffer solution (25 mM Tris–HCl–/0.1
M
Na2SO4) at pH 8.7. The serum was loaded on the affinity
gel and washed with buffer solution (25 mM Tris–
HCl/22 mM Na2SO4 at pH 8.7). The hCA I isoenzyme was
eluted by buffer solution (1.0 M NaCl/0.25 M sodium
phosphate at pH 6.3). On the other hand, hCA II isoen-
zyme was eluted by another buffer solution (0.1 M sodium
acetate/0.5 M NaClO4 at pH 5.6). Both isoenzymes were
taken from the column in fractions of 2 ml.[34] All work
was carried out at 4ꢀC. The hCA isoenzymes activity were
measured by following the change in specific absorbance
(348 nm) of p-nitrophenylacetate to p-nitrophenolate ion
over a period of 3 min at room temperature (25ꢀC) using
a spectrophotometer (Thermo Scientific, UV–vis spectro-
photometer) according to the method of Verpoorte
et al.[35] There was 0.4 ml of 0.05 M Tris–SO4 buffer
(pH 7.4), 3 ml of 3 mM p-nitrophenylacetate, 0.2 ml of
H2O and 0.1 ml of enzyme solution in the test tube con-
tent of this reaction.[36] Esterase activity assays were
identified from a series of experiments at three different
novel pyrazoles derivatives (Py1–8).
2.4.11 | 1,3-bis(3-Chloro-2-
hydroxypropyl)-5-methyl-4,6-bis(p-tolyl)-
1,3,5-triazinane-2-thione (2d)
5-Methyl-4,6-di(p-tolyl)-1,3,5-triazinane-2-thion (1b)
(0.01 mol) was dissolved in a 2:1 ratio of acetylacetone
and ethyl alcohol (10:5 ml). Then, epichlorohydrin
(0.26 ml, 3.3 mmol) was added drop by drop. After dis-
solving in the stirrer for 25 min, 0.03 g AlCl3 catalyst was
added and mixed by heating at 65–70ꢀC. After determin-
ing the full completion of reaction, the solution was evap-
orated and cleansed in ethyl alcohol solution.
2.5.2 | AChE inhibition study
1H NMR (300 MHz, DMSO-d6, δ): d (mixture of two
diastereoisomers) = 2.19 s (3 H, CH3), 2.27 s (3H, CH3N),
2.45 (t, J = 11.75 Hz, 1 H, NH), 3.40, 3.65, (d,
J = 11.75 Hz, 2H, CH2), 3.47, 3.72 (d, J = 8.5 Hz, 2H,
CH2), 3.77 [d 1H, CH (OH)], 5.01 (s, J = 8.5 Hz, 1H, CH),
4.81 (m, 1H, OH), 6.9 (m, 8H, CH–Ar).
The inhibitory effects of novel N-substituted triazinane-
2-thiones derivatives (1a–g and 2a–d) on AChE activity
were performed according to the spectrophotometric
method of Ellman et al.[37] Acetylthiocholine iodide
(AChI) substrate was utilized for the inhibition act and
enzymatic reaction. 5,50-Dithio-bis(2-nitro-benzoic) acid
(DTNB) compound was utilized in this study for the
13C NMR (75 MHz, DMSO-d6): d (mixture of two
diastereoisomers) = 24.3 (CH3–Ph), 33 (CH3–N-), 49.5 (-

6 of 16
SUJAYEV ET AL.
measurement of the AChE activity. Briefly, 750 μL of
sample solution dissolved in deionized water at different
concentrations with 100 μL of Tris/HCl buffer (1.0 M,
pH 8.0) and 50 μL of AChE solution were mixed and
incubated for 64 min at 21ꢀC.[38,39] Then reaction was ini-
tiated by the addition of 50 μL of AChI. Then 50 μL of
DTNB (0.5 mM) was added. The hydrolysis of these sub-
strates was monitored spectrophotometrically by forma-
tion of the yellow 5-thio-2-nitrobenzoate anion as the
result of the reaction of thiocholine with DTNB, released
by enzymatic hydrolysis of AChI, with an absorption
maximum at a wavelength of 412 nm.[40–45] For the com-
putation of Ki values of these compounds, three diverse
novel N-substituted triazinane-2-thiones derivatives (1a–
g and 2a–d) concentrations were utilized. One AChE
enzyme unit is the amount of enzyme that hydrolyzes
1.0 mol of AChI to choline and acetate per minute at
pH 8.0 at 37ꢀC. Finally, the Lineweaver–Burk[46] curves
were drawn.
their various transformations were investigated.
Thus, 5-(1,5-diaminopentan-3-yl)-4,6-dihydrochloride is a
result of the thiourea trifluorous acetic acid catalyst
reaction in combination with benzaldehyde, salicylic
aldehyde, anisilaldehyde and three-component condensa-
tion at
a single stage with various amines, bis
(2-hydroxyphenyl)-1,3,5-triazinane-2-thione (1a), 5-me-
thyl-4,6-di(p-tolyl)-1,3,5-triazinane-2-thione (1b), 5-ethyl-
4,6-bis(2-hydroxyphenyl)-1,3,5-triazinane-2-thione (1c),
4,6-diphenyl-1,3,5-triazinane-2-thione
(1d),
5-ethyl-
4,6-di(p-tolyl)-1,3,5-triazinane-2-thione (1e), 5-methyl-4,-
6-bis(4-methoxyphenyl)-1,3,5-triazinane-2-thione (1f) and
60–70%
yield-effective
synthesis
of
5-ethyl-4,-
6-bis(4-methoxyphenyl)-1,3,5-triazinane-2-thione (1 g)
(Scheme 1).
At the next stage, reactions of triazinanes with nucle-
ophilic reagents were investigated. New compounds
(2a–d) which are not known in the literature were
synthesized from interaction of 5-methyl-4,6-bis(p-tolyl)-
1,3,5-triazinane-2-thione (1b) with 1,2-epoxybutane,
1,2-epoxypropane, 3-chlor-butanol-1 and 1,2-epoxy-
3-chloropropane as nucleophilic reagents (Scheme 2).
The physical–chemical constraints and output of synthe-
sized compounds (1a–g and 2a–d) are shown in Table 1.
2.5.3 | Measurement of α-glycosidase
inhibitory activity
α-Glycosidase inhibitory efficacy of novel N-substituted
triazinane-2-thiones derivatives (1a–g and 2a–d) was
performed using p-nitrophenyl-D-glycopyranoside (p-
NPG) as the substrate, according to the procedure of
Tao et al.[47] Samples were prepared by dissolving
10 mg in 10 ml (EtOH: H2O). First, 100 μL of phos-
phate buffer was mixed with 30 μL of the enzyme
solution in phosphate buffer (0.15 U/ml, pH 7.4) and
30–200 μL of the sample.[48] Multiple solutions in
phosphate buffer were prepared in case of full enzyme
inhibition. Then it was preincubated at 30ꢀC for
10 min prior to adding the p-NPG for the initiation of
the reaction. Also, 50 μL of p-NPG in phosphate buffer
(5 mM, pH 7.4) after preincubation was added and
again incubated at 37ꢀC. The absorbances were spec-
trophotometrically measured at 405 nm.[49,50] The IC50
amount was calculated from activity (%) vs. plant con-
centration plots. Lineweaver–Burk graphs were used to
determine Vmax and other inhibition parameters. The
Ki was calculated from these graphs.
3.2 | Computational investigation
We computed a model reaction of Scheme 1 which yields
selectively 4,6-diphenyl-1,3,5-triazinane-2-thione (1d).
The computations were performed by using Discrete
fourier transform (DFT) with the B3LYP functional and
6-31G*[51] basis sets for H, C, N, F and O. The 6–31+
+G(d,p) basis was used for sulfur and chlorine in the
catalyst (CX3COOH) according to recent recommenda-
tions.[52] Other (e.g. WB97XD, M06 L) along with the
3 | RESULTS AND DISCUSSION
3.1 | Chemistry
At the first stage, new cyclic compounds were synthe-
sized on the basis of thiourea amines and triple-
component condensation with salicylic aldehyde and
SCHEME 1 Derivatives of triazinanethione (1a–1g)

SUJAYEV ET AL.
7 of 16
SCHEME 2 Derivatives of triazinane-thione (2a–2d)
same basis sets was tested for the optimization of inter-
mediate and transition state structures. In particular,
sulfur-containing structures could not be optimized.
Calculations show that B3LYP is better at describing
delocalization in the thiourea structure. The Gaussian
09 package was used for all calculations.[53] The reaction
TABLE 1 Physico-chemical characteristics of derivatives of triazinane-thione
Found/calculated (%)
Number
1a
Compounds
Tm.p. (ꢀC)
175–176
Brutto Formula
C
H
N
S
Yield (%)
58:74
58:76
6:68
6:70
18:04
18:06
8:25
8:26
C19H26O2N5S
81
69:42
69:45
6:73
6:75
10:82
10:84
10:29
10:27
1b
166–168
C18H21N3S
76
(Continues)

8 of 16
SUJAYEV ET AL.
TABLE 1 (Continued)
Found/calculated (%)
Number
1c
Compounds
Tm.p. (ꢀC)
Brutto Formula
C
H
N
S
Yield (%)
61:98
62:01
5:75
5:78
12:76
12:77
9:73
9:75
177
C17H19O2N3S
89
66:89
66:92
5:53
5:58
15:48
15:51
11:75
11:90
1d
112
C15H15N3S
85
70:12
70:15
7:04
7:08
12:92
12:93
9:85
9:86
1e
192–194
C19H23N3S
89
62:94
62:97
6:09
6:12
12:24
12:26
9:33
9:34
1f
178
C18H21O2N3S
77
63:85
63:86
6:41
6:44
11:76
11:78
8:96
8:99
1 g
179–181
C19H23O2N3S
70
(Continues)

SUJAYEV ET AL.
9 of 16
TABLE 1 (Continued)
Found/calculated (%)
Number
2a
Compounds
Tm.p. (ꢀC)
Brutto Formula
C
H
N
S
Yield (%)
68:91
68:93
7:53
7:57
10:97
10:99
8:35
8:37
205
C22H29ON3S
65
68:24
68:29
7:29
7:32
11:38
11:41
8:67
8:69
2b
197
C21H27 ON3S
86
68:88
68:93
7:27
7:32
10:97
10:98
8:35
8:36
2c
200–202
C22H29ON3S
68
62:42
62:45
6:39
6:44
10:01
10:03
7:63
7:65
2d
195–196
C21H26ON3SCl
64
path was calculated both for experimental reaction condi-
tions (1 atm, 313.15 K), and for 1 atm, 373 K to see the
effect of elevated temperature on energy barriers. The sol-
vent effect was calculated with a self-consistent reaction
field continuum solvation model (with a dielectric con-
stant for water). Optimized Cartesian coordinates, total
energies, Gibbs energies and enthalpies of all structures
were calculated.
The over-the-counter (OTC) synthesis mechanism of
4,6-diphenyl-1,3,5-triazinane-2-thione (1d) was initiated
with ammonia benzaldehyde interaction (Scheme 3). We
previously investigated and scrutinized theoretically the
interaction of small molecules as a starting point of a
reaction in multicomponent synthesis.[54] The amine
group of this intermediate reacted further with additional
benzaldehyde molecule. After the dihydroxylation step

10 of 16
SUJAYEV ET AL.
SCHEME 3 Quantum chemical
calculation based mechanism of CX3COOH
catalyzed OTC condensation reaction (1d)
(TS4) the thiourea amine group attacked the olefinic car-
bon of the intermediate (see TS5). Further
dihydroxylation (TS6) and proton abstraction (TS7)
resulted in the product (4,6-diphenyl-1,3,5-triazinane-
2-thione) formation. We calculated the reaction
(Scheme 1) cycle three times, changing the hydrogen
atoms of the methyl group in AA with fluorine and chlo-
rine atoms. The potential energy surface diagram in
Figure 1 includes whole three cycles of the OTC mecha-
nism with AA, TFA, and TCA catalysts. The computation
shows that there is not a major difference in energy bar-
riers (see energy span for the routes) for AA and TFA
catalyzed routes (grey and blue in Figure 1). Surprisingly,
TCA minimizes energy barriers considerably compared
with the AA and TFA.
Structural analysis of the initial three transition
states of Figure 1 clearly shows how halogen atoms
affect energy barriers. As seen from the F-TS1 and
Cl-TS1, the catalyst O–H bonds (1.08 and 1.10 Å) are
elongated compared with the AA (1.04 Å) case
(H-TS1). Replacement of the methyl hydrogen atoms
with halogen atoms in AA facilitates proton transfer via
the elongated bonds to the oxygen of the benzaldehyde
carbonyl and as a result decreases the energy barrier by
FIGURE 2 The optimized
structures of F-TS1, F-TS2, F-TS3, Cl-
TS1, Cl-TS2, Cl-TS3, H-TS1, H-TS2 and
H-TS3 with important bond lengths
(given in Å) and angles (deg). Phenyl
groups are omitted for clarity

SUJAYEV ET AL.
11 of 16
about 6 kcal/mol. The benzaldehyde interaction with
azanediylbis (phenylmethanol; F-INT2, Cl-INT2, and
H-INT2) requires higher activation energy (F-TS2
25.2 kcal/mol and Cl-TS2 27.2 kcal/mol) in the case of
halogenated acetic acids for INT2TS2. AA acts as a
reasonable catalyst, here rendering proton transfer to
the carbonyl oxygen via 23.4 kcal/mol energy barrier.
AA is also a superior catalyst for the dialkylammonium
24 kcal/mol). The smallest activation energy was seen
with the H-TS5 structure. As seen from Figure 3 the AA
carbonyl oxygen binds to proton (-NH-) of the intermedi-
ate via hydrogen bonding (1.88 Å), which serves as a sup-
porter for the acid for easy proton abstraction from the
thiourea amine group. The last hydroxyl abstraction gen-
erates an intermediate for the ring closure. The hydroxyl
abstraction occurs via the smaller energy barriers (almost
barrierless) for TCA (Cl-TS6) and TFA (F-TS6) acid-
catalyzed routes. This route was calculated to be
12 kcal/mol energetic in case of AA, despite structural
orientations (e.g. bond lengths, angles and hydrogen
bonding) being quite similar to those in the halogenated
catalysts case. The nucleophilic attack from the thiourea
amine group to the sp2 hybridized carbon (hydroxyl
group from the carbon at the previous step) yielded the
product formation via ring closure (see Figure 4).
TS7 in the three cases (grey, blue, and orange) were
concerted transition states because of proton abstraction
from the thiourea amine group and the ring closure
occurring at the same time. The activation energies were
22.2, 20.4 and 19.3 kcal/mol, respectively, for F-TS7, H-
TS7 and Cl-TS7. Overall reaction was calculated to be
33.4 kcal/mol endergonic reaching the product as a trans
conformer.
(-NH2+-,
H-INT4)
deprotonation
(H-TS3,
ΔG = 0.1 kcal/mol). It can be rationalized with the AA
binding to the structure (see Figure 2, H-TS3) via
hydrogen bonding (1.68 Å) between the hydroxyl and
the acetic acid carbonyl group. This kind of design of
the structure is not seen in analogous halogen-bearing
transition states (F-TS3 and Cl-TS3). In these transition
states, the carbonyl of the catalyst and the hydroxyl
cannot be prone to hydrogen bonding because of the
low electron density (electron density flows toward hal-
ogen atoms) on the carbonyl oxygen. At the next stage,
one of the hydroxyl groups attached to the benzylic car-
bons was removed via dehydration (F-, Cl- and H-TS4).
Catalytic performance of the AA declines at the next
step, dehydration energy barrier (H-TS4), was calcu-
lated to be 10.4 kcal/mol, which is 7.7 kcal F-TS4, and
7.3 kcal higher than the Cl-TS4 energy barrier. The
thiourea (INT6TS5) and benzaldehyde (INT2TS2) addi-
tions to the cycle were calculated to have the highest
energy barriers, and can be accepted as rate-limiting
steps for all three cycles (grey, orange and blue).
At the reference, solvent-induced conversion of the
cis conformer to trans in a polar solvent and the stability
of the trans conformer were studied by XRD and NMR
analysis. Our calculations are in good agreement with
the experimental observations showing that the
trans conformer (4,6-diphenyl-1,3,5-triazinane-2-thione)
is stable in the water medium. The cis conformer could
The activation energy of TFA-catalyzed thiourea addi-
tion (F-TS5, 26.8 kcal/mol) was greater than that for
TCA (Cl-TS, 25.9 kcal/mol) and AA (H-TS5,
FIGURE 3 The optimized structures of F-
TS4, F-TS5, F-TS6, Cl-TS4, Cl-TS5, Cl-TS6, H-
TS4, H-TS5 and H-TS6 with important bond
lengths (given in Å) and angles (deg). Phenyl
groups are omitted for clarity

12 of 16
SUJAYEV ET AL.
FIGURE 4 The optimized structures of F-
TS7, Cl-TS7, and H-TS7 with important bond
lengths (given in Å) and angles (deg). Phenyl
groups are omitted for clarity
not be allocated for the TS7. Moreover, the trans
arrangement of the product phenyl groups started from
TS3 and further optimized transition states and
intermediates.
and 5-ethyl-4,6-bis(p-tolyl)-1,3,5-triazinane-2-thione (1e)
were obtained as excellent hCA I inhibitors with values
of Ki of 13.44 4.39 and 30.36
3.45 nM, respectively.
The hCA I inhibition effects of all novel N-substituted
triazinane-2-thiones derivatives (1a–g and 2a–d) were
found to be greater than that of acetazolamide, which
was a clinically standard CA inhibitor.
3.3 | Pharmacological results
Against the physiologically dominant isoform hCA II,
novel N-substituted triazinane-2-thione derivatives (1a–g
and 2a–d) demonstrated values of Ki varying from
The cytosolic hCA I, and II isoforms are ubiquitous and
both can be used for treatment of diseases (e.g. as anti-
glaucoma or diuretics factors) and off-targets.[55] Also,
the membrane-associated hCA IV isoenzyme is a drug
used for multiple pathologies, like retinitis pigmentosa,
glaucoma (together with hCA II and XII), rheumatoid
arthritis and stroke.[56] Presently, there are 22 clinically
utilized CA inhibitor compounds, but all of them are
show little or no isoenzyme specificity.[57] Because if this,
there is a need to develop CA inhibitor compounds that
are isoenzymes selective, for instance, the targeted inhibi-
tion action of CA IX isoform can be used for anticancer
chemotherapy.[58] The design of CA isoform-specific
inhibitor compounds is challenging owing to the high
sequence similarity within the active sites and structural
homology of the active CA isoenzymes. Additionally, we
report the inhibition effect of novel N-substituted
triazinane-2-thiones derivatives (1a–g and 2a–d) on two
catalytically active isoforms, hCA I and II, as well as
against AChE and α-glycosidase enzymes. The enzymes
inhibition activity data of novel N-substituted triazinane-
2-thiones derivatives (1a–g and 2a–d) reported here are
shown in Table 2, and the following comments can be
drawn from these data.
10.41
4.8 to 72.6
17.66 nM (Table 2). These com-
pounds had high inhibition effects toward hCA II. On the
other hand, the standard compound acetazolamide
showed Ki of 106.73
bis(2-Hydroxypropyl)-5-methyl-4,6-bis(p-tolyl)-
1,3,5-triazinane-2-thione (2b) and 4,6-diphenyl-
20.73 nM against hCA II. 1,3--
1,3,5-triazinane-2-thione (1d) showed the most inhibition
effect with Ki values of 10.41 4.8 and 13.66 2.36 nM,
respectively.
Recently, many synthetic or natural compounds and
their derivatives were discovered as a novel potential
therapies for the treatment of AD.[59] Followed by these
evaluations, classes of synthetic and natural products
derivatives were synthesized and investigated for AChE
enzyme inhibitory activity in our laboratory.[60] In our
study, novel N-substituted triazinane-2-thione derivatives
(1a–g and 2a–d) were investigated for their ability to
inhibit AChE, which was the primary ChE in the body.
According to our data, inhibitory results of these mole-
cules revealed a significant elevation in the case of AChE.
Considering the results, all molecules expressed
appreciably higher inhibition activity. All of these deriva-
tives had significantly higher AChE inhibitory activity
than standard AChE inhibitors such as tacrine. The Ki
values of these compounds and standard compound
(tacrine) are summarized in Table 2. As can be seen from
the results obtained, these compounds effectively
inhibited AChE, with Ki values in the range of
36.82 9.95 to 108.48 1.17 nM. However, all of these
compounds had similar inhibition profiles. The most
active 5-ethyl-4,6-bis(2-hydroxyphenyl)-1,3,5-triazinane-
2-thione (1c), 5-ethyl-4,6-bis(p-tolyl)-1,3,5-triazinane-
2-thione (1e) and 5-methyl-4,6-bis(4-methoxyphenyl)-
The results presented in Table 2 indicate that novel
N-substituted triazinane-2-thiones derivatives (1a–g and
2a–d) had an effective inhibition profile against slow
cytosolic hCA I isoform. The hCA I isoform was inhibited
by these compounds at low nanomolar levels, the Ki of
which differed between 13.44 4.39 and 74.98 6.25 nM.
On the other hand, acetazolamide, considered a broad-
specificity CA inhibitor owing to its widespread inhibi-
tion of CAs, showed a Ki value of 120.36
against hCA isoenzyme. Among the inhibitors,
5-methyl-4,6-bis(p-tolyl)-1,3,5-triazinane-2-thione (1b)
15.85 nM
I

SUJAYEV ET AL.
13 of 16

14 of 16
SUJAYEV ET AL.
1,3,5-triazinane-2-thione (1d) showed Ki values of
36.82 9.95, 40.28 9.70 and 42.82 9.72 nM, respec-
tively. Three out of the four drug compounds newly
approved for the therapy of AD are based on the inhibi-
tion of AChE enzyme; hence, their usefulness is as yet
limited since the long-term (for instance, >6 months)
efficacy and safety are not entirely clear. Thus, the
development of efficient bioanalytical procedures for the
analysis of AChEIs in biological samples is important in
assessing the pharmacokinetics and bioavailability of
these drugs as well as related compounds, metabolites
and degradation products.
hyperglycemia.[65] For the α-glycosidase enzyme, the
novel N-substituted triazinane-2-thiones derivatives
(1a–g and 2a–d) had IC50 values in the range of
581.44–1083.20 nM and Ki values in the range of
624.62
100.34–1124.16
205.14 nM (Table 2 and
Figure 5). The results clearly recorded that all these
derivatives (1a–g and 2a–d) demonstrated efficient
α-glycosidase inhibitory effects compared with acarbose
(IC50: 22.8 μM)[66,67] as a standard α-glycosidase inhibitor.
Indeed, the most effective Ki values were obtained
by 5-ethyl-4,6-bis(p-tolyl)-1,3,5-triazinane-2-thione (1e)
and 4,6-diphenyl-1,3,5-triazinane-2-thione (1d), with Ki
Postprandial glucose rates can be adjusted via
α-glycosidase inhibitor compounds. Inhibition of this
enzyme delays and in some cases halts carbohydrate
digestion, thus prolonging overall carbohydrate digestion
time, causing a reduction in the rate of glucose absorp-
tion and accordingly decreasing postprandial plasma
glucose rise.[61] Currently, α-glycosidase inhibitor
compounds like miglitol, acarbose and voglibose are oral
blood glucose-lowering drugs that are generally uti-
lized.[62] They are also the only drug class that does not
pose a pathophysiological issue in type 2 diabetes
melitis.[63] They reduce postprandial hyperglycemia
without inducing insulin secretion; these compounds do
not induce hypoglycemia and have a good safety profile,
although gastrointestinal adverse effects may limit
long-term compliance with therapy.[64] The research for
the new group of factors from natural resources,
especially from traditional medicines, has become an
attractive approach for the treatment of postprandial
values of 624.62
respectively.
100.34 and 649.15
88.81 nM,
Animal research has revealed that glucose may
improve memory through a facilitation of acetylcholine
synthesis and release in the brain. This glucose-related
memory improvement has prompted research in elderly
humans. These studies have shown that the memory-
improving action of glucose depends on each individual's
blood glucose regulation. Based on these data,
researchers have evaluated the effect of glucose on
memory in patients with AD. DM contributes to cogni-
tive impairment in the elderly, and constitutes a risk fac-
tor for dementia, possibly including Alzheimer's disease.
The development of effective synthetic α-glycosidase
inhibitors may prove important for antidiabetic chemo-
therapy and treatments for other related disorders. These
compounds are also excellent carbonic anhydrase inhibi-
tors, making them interesting antiglaucoma drug
candidates.
FIGURE 5 Determination of
Lineweaver–Burk graphs for excellent
inhibitors of hCA I (1b), hCA II (1b),
acetylcholinesterase (AChE) (1c) and α-
glycosidase (1d) enzymes

SUJAYEV ET AL.
15 of 16
[14] H. I. Gul, A. Demirtas, G. Ucar, P. Taslimi, I. Gulcin, Lett.
Drug Des. Discov. 2017, 14, 573.
4 | CONCLUSIONS
[15] Ç. Bayrak, P. Taslimi, I. Gulcin, A. Menzek, Bioorg. Chem.
2017, 72, 359.
[16] A. Aktas¸, P. Taslimi, I. Gulcin, Y. Gök, Arch. Pharm. 2017,
350, e1700045.
[17] P. Taslimi, A. Sujayev, E. Garibov, N. Nazarov, Z. Huyut,
S. H. Alwasel, I. Gulcin, J. Biochem. Mol. Toxicol. 2017, 31,
e21897.
[18] N. Öztaskın, P. Taslimi, A. Maras, S. Göksu, I. Gulcin, Bioorg.
Chem. 2017, 74, 104.
[19] P. Taslimi, S. Osmanova, I. Gulcin, S. Sardarova,
V. Farzaliyev, A. Sujayev, R. Kaya, F. Koc, S. Beydemir,
S. H. Alwasel, O. I. Kufrevioglu, J. Biochem. Mol. Toxicol. 2017,
31, e21931.
Novel N-substituted triazinane-2-thiones derivatives (1a–
g and 2a–d) were used in the present paper and also
recorded efficient inhibition profiles against AChE, hCA
I and II, and α-glycosidase enzymes. In this paper,
nanomolar levels of IC50 values were obtained for all
derivatives on these metabolic enzymes. Thus, these com-
pounds can be selective inhibitors of α-glycosidase and
AChE enzymes as anticholinergic and antidiabetic drugs
potentials.
CONFLICT OF INTEREST STATEMENT
The authors declare that there are no conflicts of interest.
ꢀ
[20] U. M. Koçyigit, P. Taslimi, H. Gezegen, I. Gulcin, M. Ceylan,
J. Biochem. Mol. Toxicol. 2017, 31, e21938.
[21] I. Gulcin, M. Abbasova, P. Taslimi, Z. Huyut, L. Safarova,
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ORCID
Parham Taslimi https://orcid.org/0000-0002-3171-0633
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How to cite this article: Sujayev A, Taslimi P,
Kaya R, et al. Synthesis, characterization and
biological evaluation of N-substituted triazinane-2-
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