Conventional and Microwave-assisted Total Synthesis, Antioxidant Capacity, Biological Activity, and Molecular Docking Studies of New Hybrid Compounds

Article abstract of DOI:10.1002/jhet.2760

Thiomorpholine was converted to the corresponding 1,3,4-oxadiazole (4), arylidenehydrazide (5a, 5b, 5c, 5d, 5e), and 1,2,4-triazole (7a and, 7b) derivatives via the formation of hydrazide (3). Compounds 4 and 7 were next converted to the corresponding Man

Full text of DOI:10.1002/jhet.2760

Month 2016
Conventional and Microwave-assisted Total Synthesis, Antioxidant
Capacity, Biological Activity, and Molecular Docking Studies of New
Hybrid Compounds
a
b
c
d
e
b
e
Serpil Demirci, * Arif Mermer, Gokhan Ak, Fatma Aksakal, Nesrin Colak, Ahmet Demirbas, Faik Ahmet Ayaz, and
b
Neslihan Demirbas
a
Department of Crop Production and Technology, Bulancak Kadir Karabas School of Applied Science, Giresun
University, 28000, Giresun, Turkey
Department of Chemistry, Karadeniz Technical University, 61080, Trabzon, Turkey
Department of Biology, Recep Tayyip Erdoğan University, 53100, Rize, Turkey
Department of Chemistry, Gebze Technical University, 41400, Kocaeli, Turkey
b
c
d
e
Department of Biology, Karadeniz Technical University, 61080, Trabzon, Turkey
*
E-mail: demirciserpil17@gmail.com
Additional supporting information may be found in the online version of this article.
Received March 5, 2016
DOI 10.1002/jhet.2760
Published online 00 Month 2016 in Wiley Online Library (wileyonlinelibrary.com).
Thiomorpholine was converted to the corresponding 1,3,4-oxadiazole (4), arylidenehydrazide (5a–e), and
,2,4-triazole (7a and, 7b) derivatives via the formation of hydrazide (3). Compounds 4 and 7 were next
1
converted to the corresponding Mannich bases containing piperidin, β-lactam, fluoroquinolone, piperazine,
or morpholine core. Conventional and microwave-assisted methods were used for all syntheses. The effect
of acid catalyst on Mannich reactions was also investigated. All the newly synthesized compounds were
screened for their antimicrobial, antiglucosidase, antilipase, anti-urease, and antioxidant activities. Most
exhibited good–moderate antibacterial and/or antifungal activity. Docking of some of the synthesized com-
pounds into the active sites of lipase, α-glucosidase, and urease was carried out in order to predict the binding
affinities and noncovalent interactions stabilizing the enzyme–ligand complexes. Docking results
complemented well the experimental results on inhibitory effects of compounds. Higher binding affinities
were observed for active compounds in contrary to inactive ones.
J. Heterocyclic Chem., 00, 00 (2016).
INTRODUCTION
Morpholine and thiomorpholine moieties are important
structural units present in various biologically active
heterocyclic compounds because of their favorable
lipophilicity and hydrophilicity [10]. For instance,
sutezolid (Fig. 1), from the oxazolidinone class
antibiotics in phase II clinical trials, is converted to
sulfone and sulfoxide metabolites.
An antioxidant is a molecule that inhibits the oxidation
of other molecules from free radicals that can initiate
chain reactions. Antioxidants terminate these chain
reactions by removing free radical intermediates and
inhibit other oxidation reactions. Antioxidants are
commonly added to food products such as vegetable oils
and prepared foods in order to prevent or delay their
deterioration because of the action of the air.
Antioxidants, which potentially reduce the risk of cancer,
significantly slow the progression of age-related macular
degeneration. A number of methods for assessing
antioxidant capacity that are adaptable for measuring
compounds with lipophilic and hydrophilic antioxidant
properties have been developed [11].
During the past few years, the rapid proliferation of
drug-resistant bacteria originating from the excessive and
inappropriate use of commonly used antibiotics has
become
a serious problem in hospitals and the
community in general [1–6]. This rapid emergence of
drug-resistant pathogens that reduce the efficacy of
commonly used antibiotics underscores an urgent need
to discover and develop new antimicrobial agents with
different structures to the existing ones that act via novel
mechanisms. Another approach for the development of
new nonresistant antibiotics is to combine two or
more pharmacophores into a single molecule. These
synergistic antimicrobial combinations are reported to
have several major advantages, including the potential to
slow the development of drug resistance, a broader
antimicrobial spectrum, and a potential reduction in the
dose and toxicity of each drug [7,8]. Considerable focus
has also been placed on the development of methods
that reduce the toxicity of parent compounds and on the
development of new, potentially less toxic hybrid
compounds [9].
The design of more economic and eco-friendly one-pot
syntheses without hazardous solvents as well as expensive
©
2016 Wiley Periodicals, Inc.

S. Demirci, A. Mermer, G. Ak, F. Aksakal, N. Colak, A. Demirbas, F. A. Ayaz, and N. Demirbas Vol 000
protonation. This restriction of protonation results in
enhancement of absorption through biomembranes. At
the same time, the basic function of Mannich bases
renders the molecules soluble in aqueous solvents when
Figure 1. The structure of sutezolid. [Color figure can be viewed in the
online issue, which is available at wileyonlinelibrary.com.]
they are transformed into aminium salt [23–25].
In the light of this and our own research into the synthesis
of biologically active compounds, this article presents the
conventional and MW-assisted synthesis and modeling of
new thiomorpholine derivatives incorporating different
pharmacophores as hybrid molecules possessing
antimicrobial, anti-α-glucosidase, antilipase and anti-
urease activities and antioxidant capacity.
and toxic reagents has become one of the most investigated
and studied fields of synthetic organic chemistry. These
methodologies involve a combination of a number of
technologies and economic targets. Multicomponent
reactions, which contain the reaction of at least three
components via one-pot process to give a single
product, represent a unique strategy leading to the
formation of various bioactive molecules, because of their
convergence, low energy consumption, minimum waste
production, facile execution, high selectivity, and
productivity [12–15]. In addition, microwave (MW)-
assisted techniques are reported to be more effective in
terms of the environment, reaction time, high yields, ease
of work-up, and isolation of products. Moreover, solvents,
which are often expensive, toxic, and difficult to remove
in the case of aprotic dipolar solvents with high boiling
points and are environmentally polluting agents, are not
necessary for most MW-assisted synthesis [16–20].
Therefore, the combination of one-pot multicomponent
reactions and MW irradiation techniques has been a very
attractive methodology for the production of new
bioactive compounds.
RESULTS AND DISCUSSION
Chemistry.
The main aim of this article was to
synthesize new thiomorpholine derivatives containing
different heterocyclic moieties by eco-friendly way and to
investigate their antimicrobial, antiglucosidase, antilipase,
anti-urease, and antioxidant activities. Moreover,
molecular docking was carried out in order to predict the
binding mode and affinities of the newly synthesized
compounds to target enzymes. The most important
interactions between compounds and enzymes were
obtained with the docking analysis. Synthesis of the
intermediate and target compounds was performed
according to the reactions outlined in Schemes 1–3. The
synthesis of the targeted compounds was achieved by
both MW (eco-friendly) and conventional (traditional)
methods, some of which also involve the use of an acid
catalyst. In this article, MW-assisted techniques were a
more effective means of performing syntheses in terms of
the environment, reaction time, high yields, ease of work-
up, and isolation of products. Time and yield data for
The classic Mannich reaction, a one-pot three-
component reaction, leads to the formation of
aminoalkylated compounds, which are used to obtain
prodrugs of amine as well as amide-containing drugs
[
21,22]. The group linked to the parent amine by
Mannich reaction is believed to increase the lipophilicity
of molecule at physiological pH values by reducing their
2 2 2 2
Scheme 1. Reaction and conditions for the synthesis of compounds 2–6. i: BrCH CO Et in THF, TEA, 24 h, 60°C (method 1) or BrCH CO Et in THF,
TEA, 110°C, 15 min, 200 W (method 2). ii: H
CS in EtOH–H O, 15 h reflux (method 1) or 8 min 150 W MW irradiation MW (method 2). iv: Benzaldehyde (for 5a), 4-methoxybenzaldehyde (for 5b),
-pyridinecarboxaldehyde (for 5c), 3-hydroxy-4-methoxybenzaldehyde (for 5d), indol-3-carbaldehyde (for 5e), 6 h, reflux. v: Benzylisothiocyanate (for
a), phenylisocyanate (for 6b) in absolute EtOH, reflux, 18 h (method 1) or 8 min 150 W MW irradiation MW (method 2).
2 2 2
NNH .H O in EtOH, 12 h, reflux (method 1) or 7 min 100 W MW irradiation MW (method 2). iii: KOH,
2
2
4
6
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New Hybrid Compounds
Table 1
Scheme 2. E/Z geometrical isomers and cis/trans conformers in
compounds 5a–e.
Time, power, and yield data for compounds 2–4, 6a,b, 7a,b, 9a,b, and
10a,b.
Microwave irradiation method
Conventional method
Time
(min)
Power
(W)
Yield
(%)
Temp.
(°C)
Time
(h)
Yield
(%)
No.
2
3
4
15
7
8
7
7
5
7
30
30
15
10
200
100
150
120
120
200
200
200
200
150
200
88
97
93
98
85
98
88
89
97
90
92
110
80
120
80
24
12
10
18
18
3
70
50
53
62
65
75
82
40
70
75
50
6
6
7
7
9
9
1
1
a
b
a
b
a
b
0a
0b
80
150
150
150
150
150
120
3
18
18
18
12
newly synthesized compounds produced by MW and
conventional methods are given in Tables 1 and 2.
In this article, the ester (2) was obtained from the
reaction of commercially obtained thiomorpholine with
ethyl bromoacetate in dry tetrahydrofuran. The reaction
was investigated in THF under conventional heating
conditions at 60°C as well as under MW irradiation
conditions with a view to maximizing the yield of the
product and minimizing reaction time, and the reaction
was monitored by TLC. MW irradiation decreased the
reaction time from 24h to 15min and increased the
yields from 70% to 88% (Table 1). The optimum reaction
condition was assessed at 200-W maximum power in
closed vessel. The FTIR spectrum of compound 2
The treatment of 2 with hydrazine hydrate in ethanol
under reflux conditions yielded the hydrazide, 3. The
reaction was examined under MW conditions without any
solvent as well. The complete conversion of the starting
ester (2) was observed after MW irradiation at 100W for
12min. It is noteworthy to underline that shorter reaction
time or lower MW energy power caused to lower
conversion rate, while increasing reaction time or MW
power resulted in decomposition of the target product as
revealed by TLC analysis. With the use MW irradiation,
the improved yield was assessed as 97%. Compound 3
showed the disappearance of the characteristic signals
for the ethyl group and the appearance of signals at 4.22
À1
showed an absorption band at 1753 cm , corresponding
to the vibration of the carboxyl (–C=O), while the
spectrum of 3, obtained from the treatment of 2 with
hydrazine hydrate, showed the disappearance of the
(–NHNH
changing with D
proton, respectively.
2
) and 8.91 (–NHNH
) ppm (controlled by
2
O) integrating for two protons and one
2
characteristic bands of the carboxylic acid ester and the
Compound 3 was converted to 1,3,4-oxadiazole
derivative (4) by cyclocondensation with CS in the
2
À1
appearance of strong bands in the 3225 cm
region,
attributed to (–NHNH ) stretching. Proton assignments in
H NMR spectra for compound 2 showed signals at
presence of KOH. The condensation was investigated
under MW and conventional conditions. MW irradiation
was applied at different power values of 70, 100, 150, and
200 W, and the progress of reaction was monitored by
2
1
1
.18ppm (–OCH CH ) and 4.07 (–OCH CH ) ppm
2 3 2 3
integrating for three protons and two protons, respectively.
Scheme 3. Reaction and conditions for the synthesis of compounds 7–10; i: 2 N of NaOH, in EtOH : H
2
O (1:1), 3 h, reflux (method 1) or 8 min
COONa, in EtOH, 18 h, reflux (method 1) or 8 min 150 W MW
COONa, in EtOH, 18 h, reflux (method 1) or 8 min 150 W MW irradiation MW (method 2).
1
50 W MW irradiation MW (method 2); ii: H
2
SO
4
, 2 h, rt, iii: (4)-ClC
6
H
4
CH
2
Br, CH
3
irradiation MW (method 2); iv: BrCH CO Et, CH
2
2
3
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S. Demirci, A. Mermer, G. Ak, F. Aksakal, N. Colak, A. Demirbas, F. A. Ayaz, and N. Demirbas Vol 000
Table 2
Time and yield data of compounds 11–16 using conventional, conventional with catalyst, and microwave irradiation techniques.
Microwave irradiation method
Conventional method
Conventional method with catalyst
Yield (%) Yield (%)
No.
Power (W)
Time (min)
Yield (%)
Time (h)
Yield (%)
Time (h)
(with InCl
3
)
(with p-TsOH)
1
1a
100
150
100
150
100
100
100
125
100
100
125
150
150
200
125
100
125
150
5
5
6
5
5
4
5
10
5
6
5
10
7
15
5
5
45
78
57
49
53
62
65
67
59
40
45
59
80
75
78
55
65
90
12
24
24
22
22
23
24
24
21
21
24
22
23
24
22
22
24
24
36
75
56
82
78
70
75
45
25
73
30
40
70
70
62
40
58
55
3
5
10
4
53
93
68
88
94
77
98
67
69
79
60
61
77
78
85
60
70
66
64
87
85
93
97
88
89
60
55
80
55
65
79
80
80
56
74
60
1
1b
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
2a
2b
2c
2d
2e
3a
3b
3c
4a
4b
4c
5a
5b
6a
6b
6c
4
4.5
4.5
5
6
6
12
4
4.5
4.5
4
4
4.5
4.5
10
4
TLC. The complete conversion of the starting hydrazide (3)
was observed after MW irradiation at 150W for 8 min in
water–ethanol (Table 1). It is noteworthy to underline that
shorter reaction time or lower MW energy power caused
to lower conversion rate, while increasing reaction time or
MW power resulted in decomposition of the target
product as revealed by TLC analysis.
analysis results and mass fragmentation confirming their
structures.
It is well known that arylidenehydrazides may exist as
Z/E geometrical isomers about a –C=N– double bond, and
Z and E isomers may consist of their individual cis–trans
amide conformers (Scheme 2). The literature survey
revealed that compounds containing imine bonds are
present at higher percentages in dimethyl-d6 sulfoxide
solution in the form of a geometric E isomer about a
–C=N double bond, while the Z isomers can be stabilized
in less polar solvents by an intramolecular hydrogen bond
(Scheme 2) [29].
This compound (4) was characterized by the
disappearance of the –NHNH signals in the FTIR and
2
1
H NMR spectra and the presence of singlet at 14.73 ppm
due to (–SH) function confirming the cyclisation. The
À1
(
–SH) stretching band was observed at 2549 cm in the
FTIR spectrum of 4. The C-2 and C-5 carbon atoms of
the 1,3,4-oxadiazole ring resonated at 161.22 (C-5) and
In this article, the stereochemical behavior of
compounds 5a–e was investigated in dimethyl-d6
sulfoxide solution, and two sets of signals each belonging
to the individual N=CH and NH protons of the cis and
trans conformers varying between 7.82 and 8.92ppm
(N=CH) and 10.95 and 11.65 ppm (NH) were observed.
1
3
1
78.46 (C-2), respectively, in the C NMR spectrum of
compound 4. Moreover, mass spectral data and the
elemental analysis results of compound 4 were consistent
with the assigned structure.
Our research group has previously reported the
synthesis of novel imine compounds, and most of these
exhibited several biological activities including
antimicrobial, antitumor, enzyme inhibition, and so on
The change in the peak ratio with the addition of D O
2
indicated that these signals observed as two sets stem
from cis/trans conformers but not E/Z isomers.
Additional support for the formation of the targeted
compounds, 5a–e, was obtained by the appearance of
[M + 1] ion peaks at corresponding m/z values confirming
their molecular masses; these compounds produced
elemental analysis results consistent with the proposed
structures.
[
7,26–28]. As a part of our efforts aiming to obtain
bioactive hybrid molecules, we performed the synthesis
of arylidenehydrazides (5a–e) via the condensation of
compound 3 with suitable aldehydes. In the FTIR and
1
H NMR spectra of these compounds, no signal
pointing the –NH group was observed, while additional
The treatment of hydrazide (3) with alkyliso(thio)
2
signals derived from aldehyde moiety were recorded at
cyanates,
namely
benzylisothiocyanate
and
1
13
the related chemical shift values in the H and
C
phenylisocyanate, in ethanol elicited the corresponding
NMR spectra. These imines gave reasonable elemental
carbothioamides, 6a,b. Compared with conventional
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New Hybrid Compounds
thermal heating, MW irradiation decreased the reaction
time from 18h to 7 min and increased the yields from
1-(4-chlorophenyl)ethanone was achieved under reflux
and also MW irradiation conditions producing the
corresponding 1,3-thiazole derivatives (9a,b). The
optimized condition was assessed under 150W (for 10a)
and 200W (for 9a,b and 10b) of MW irradiation in
ethanol in the closed vessel (Table 1). This idea
originated from the intent to merge two bioactive
moieties, namely thiomorpholine and 1,3-thiazol(idin), in
6
2–65% to 85–99% (Table 1). Thus, MW irradiation
allowed a rapid, green, and efficient synthesis of these
carbothioamides (6a,b) (Scheme 1). In contrast to
1
13
compound 3, H and C NMR spectra of compounds 6a,
b exhibited additional signals because of carbothioamide
moiety at the relevant chemical shift values. The FTIR
spectra of compounds 6a,b displayed additional
one structure. The disappearance of one NH signal in the
À1
1
absorption bands at 1228 (C=S) or 1703 (C=O) cm
,
FTIR and H NMR (exchangeable with D O) supported
2
indicating the presence of –C=S or a second –C=O in the
structure. These carbothioamides (6a,b) displayed mass
fragmentation and elemental analysis results consistent
with their structures.
the condensation leading to the formation of compounds
1
13
10a,b. The H and C NMR spectra of compounds 9a
and 9b exhibited additional signals at the aromatic region
originating from the 4-chlorophenyl nucleus as a result of
condensation. The absorption band observed at
The acidic treatment of compounds 6a,b afforded the
corresponding 1,3,4-thiadiazoles (8a,b) in cold-room
temperature without any solvent. The identities of
À1
À1
1725 cm
(for 9a) and 1715 cm
(for 9b) was
attributed to carbonyl function on the 1,3-oxa(thio)zole
ring. Moreover, the elemental analyses and mass spectral
data of derivatives 9 and 10 were compatible with the
1
13
compounds 8a,b were confirmed by FTIR, H and
C
NMR, and mass spectral and elemental analyses.
1
The synthesis of triazoles (7a and 7b) was performed by
basic treatment of the corresponding carbothioamides in
order to merge fluorophenylenemorpholine nucleus with
suggested structures. In the H NMR spectra of
compounds 10a and 10b, the signal due to one (–NH–)
proton was detected at 9.08 and 10.08 ppm (exchangeable
5
-mercapto-1,2,4-triazol ring and to produce an
with D O).
2
intermediate for further condensations. The reaction was
investigated in ethanol–water (1:1) under reflux conditions
as well as under MW irradiation conditions with a view to
maximizing the yield of the product and minimizing the
reaction time. Reactions were monitored by TLC. Thus,
the yield of the reaction was improved to good level
The key intermediates 4 and 7a,b can be regarded as
cyclic (thio)amides, and the hydrogen atom attached to
the nitrogen atom should be appreciably liable to
participate in the Mannich condensation. The
condensation of these intermediates (4 and 7a,b) with
formaldehyde and various primary or secondary amines
thus resulted in the formation of the corresponding
Mannich base derivatives (11–16). Three methods were
used for this treatment, including conventional and MW-
assisted synthesis and the use of a catalyst (Scheme 4).
Time and yield data for the synthesis of compounds 11–16
by MW and conventional methods are given in Table 2.
In comparison with the long refluxing time, MW
irradiation provided a more efficient and greener path for
Mannich-type condensation with a relatively higher
product yield. Subsequently, Mannich reaction was
(88–98%); however, more significantly, the reaction time
for complete consumption of starting materials was
lowered from 3h with conventional heating to remarkable
5–7min using MW irradiation at 200-W maximum power.
The structural assignments of these compounds were
1
based on their elemental analysis and spectral (IR, H
13
NMR, C NMR, and Liquid Chromatography Mass
Spectrometry (LC MS)) data. The absorption bands
À1
observed at 2831 (compound 7a) or 3280cm
(
compound 7b) were attributed to an (–SH) or (–OH)
1
group, respectively. In the H NMR spectra, these protons
conducted in the presence of InCl as a Lewis acid and p-
3
resonated at 13.80 and 13.51ppm, respectively, as a D O
toluenesulfonic acid as a Brønsted–Lowry acid. The
reactions with a catalyst were faster than the reaction
with no catalyst. The results showed that Brønsted–
Lowry acid (p-TsOH) was rather more effective than
Lewis acid (InCl3), probably because of p-TsOH
facilitating the formation of an electrophilic iminium ion.
The alkylaminomethylation was provided by the
disappearance of signal for the proton on the sulfur or
oxygen atoms of compounds 4 and 7a,b. Instead, the
appearance of a signal originated from methylene linkage
attached to the nitrogen atom of the triazole or oxadiazole
2
1
3
exchangeable singlet. In the C NMR spectra, the signals
derived from triazole C-3 and C-5 carbons were recorded
at approximately 149.56ppm (C-5) and 168.73ppm (C-2)
in accordance with the literature findings [5,7,26].
The condensation of ethyl bromoacetate with
compounds
6a,b
afforded
4-oxo-1,3-thiazolidin
derivatives (10a,b). Compared with conventional thermal
heating, MW irradiation decreased the reaction time from
1
2h to 10–15min and increased the yields from 50–75%
to 90–92% (Table 1). Thus, MW irradiation allowed a
rapid, green, and efficient synthesis of these 4-oxo-1,3-
thiazolidin (10a,b). On the other hand, the
cyclocondensation of compounds 6a,b with 2-bromo-
1
13
nucleus. Moreover, in the H and C NMR spectra,
additional signals corresponding to amine moiety used in
Mannich condensation were recorded at the relevant
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S. Demirci, A. Mermer, G. Ak, F. Aksakal, N. Colak, A. Demirbas, F. A. Ayaz, and N. Demirbas Vol 000
Scheme 4. Synthetic pathway for the preparation of Mannich bases. Suitable amine in THF, formaldehyde (HCHO), 3 h, rt (method 1); suitable amine in
THF, HCHO, 8 min 150 W MW irradiation MW (Table 2) (method 2) or suitable amine in DMF, HCHO, with an acid catalyst (Table 2) (method 3).
chemical shift values. Elemental analyses were consistent
11a–c, hybrid molecules consisting of a thiomorpholine–
azole–piperidin ring, displayed an activity on Escherichia
coli, a Gram-negative, facultatively anaerobic bacterium;
Yersinia pseudotuberculosis, a Gram-negative bacterium;
Staphylococcus aureus; Bacillus cereus, a Gram-positive
spore bacillus; and M. smegmatis, with MIC values
between 62.5 and 3.8 μg/mL. Compounds 12a–e, which
contain a norfloxacine or ciprofloxacin skeleton attached
to a thiomorpholine nucleus via an azole linkage,
exhibited excellent activities against the test
microorganisms, except for the yeast-like fungi Candida
albicans and Saccharomyces cerevisiae. Indeed, the MIC
values of these compounds varied between 0.24 and
15.8 μg/mL and are better than those of the standard drugs
ampicillin and fluconazole. Among the fluoroquinolone
with the assigned structures for these Mannich bases
+
(
[
11–16), and the mass spectra of these revealed [M] ,
+
+
+
+
M + 1] , [M + 2] , [M + Na] , and/or [M + K] ion peaks
at the corresponding m/z values, which match their
molecular formulate.
Biological activity
Antimicrobial activity.
All the newly synthesized
compounds were screened for their antimicrobial activity.
Only positive results are presented in Table 3. This
reveals that hydrazide (3), oxadiazole (4), carbo(thio)
amides (6a and 6b), and 1,3-thiazole derivatives are
superior in inhibiting the growth of Mycobacterium
smegmatis, a nonpigmented rapidly growing atypical
tuberculosis factor, with the minimal inhibition
concentration (MIC) values varying between 6 and
derivatives, compound 12e,
a
thiomorpholine–
31.3μg/mL. Among the Mannich bases, compounds
oxadiazole–ciprofloxacin hybrid, exhibited activity on
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Month 2016
Antioxidant Capacity, Biological Activity, and Molecular Docking Studies of
New Hybrid Compounds
Table 3
Screening for antimicrobial activity of the compounds (only positive results were presented) (μg/μL).
Microorganisms and minimal inhibition concentration (MIC)
No.
Ec
Yp
Pa
Ef
Sa
Bc
Ms
Ca
Sc
3
4
6
6
9
9
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
<0.24
<0.24
<0.23
0.24
<0.24
—
—
—
—
—
500
500
500
—
—
—
—
—
—
<0.24
<0.24
<0.23
0.12
<0.24
—
—
500
—
—
500
—
6
31.3
8
—
62.5
—
—
—
—
—
—
—
—
—
—
—
15.8
137
125
125
29.5
29.5
31.3
137
125
—
—
31.3
—
—
—
—
—
—
—
—
—
—
—
<0.24
68
62.5
137
29.5
29.5
29.5
68
31.3
95.3
128
—
a
b
a
b
—
—
8
500
500
8
8
8
<0.24
<0.24
<0.23
0.12
<0.24
—
—
—
—
—
—
550
—
47.6
256
—
125
125
15
3.8
3.8
<0.24
<0.24
<0.23
0.12
15.8
—
31.3
31.3
>20
>20
31.3
0.48
<0.24
<0.23
0.24
<0.24
31.3
31.3
31.3
0.46
0.46
0.46
17.2
31.25
—
—
10
—
10
1
1a
1b
1c
1
1
30.5
62.5
<0.24
<0.24
<0.23
0.03
<0.24
62.5
62.0
64.1
30.5
59.0
59.0
—
30.5
30.5
<0.24
<0.24
<0.23
0.06
<0.24
62.5
62.5
64.1
30.5
59.0
59.0
—
12a
12b
12c
12d
12e
13a
13b
13c
14a
14b
14c
15a
15b
16a
16b
16c
—
—
—
—
250
236
236
137
62.5
47.6
32.0
62.5
>128
250
236
256
—
62.5
47.6
32.0
—
—
137
125
190.6
—
—
18
62.5
190
64.1
62.5
10
62.5
190
64.1
250
10
—
62.5
—
—
Amp.
Str.
10
15
4
Flu
<8
<8
Ec, Escherichia coli ATCC 25922; Yp, Yersinia pseudotuberculosis ATCC 911; Pa, Pseudomonas aeruginosa ATCC 27853; Sa, Staphylococcus aureus
ATCC 25923; Ef, Enterococcus faecalis ATCC 29212; Bc, Bacillus cereus 702 Roma; Ms, Mycobacterium smegmatis ATCC 607; Ca, Candida albicans
ATCC 60193; Saccharomyces cerevisiae RSKK 251; Amp., ampicillin; Str., streptomycin; Flu., fluconazole; —, no activity.
C. albicans and S. cerevisiae, with MIC values ranging
from 0.24 to 15.8 μg/mL.
Pancreatic lipase inhibition.
All compounds were
evaluated with regard to pancreatic lipase activity. Some
exhibited antilipase activities at various concentrations.
The results obtained are shown in Table 4. Among the
tested compounds, 15a and 13a, which contain a
penicillanic acid (13a) or methoxyphenylpiperazine (15a)
nucleus linked to a thiomorpholinomethyl-4,5-dihydro-
The Mannich bases 13a–c and 14a–c, which contain a
β-lactam nucleus instead of fluoroquinolones in contrast
to 12a–e, exhibited activity toward E. coli, Y.
pseudotuberculosis, M. smegmatis, C. albicans, and S.
cerevisiae. Compounds 14a–c, as cephalosporin
derivatives, exhibited slight activity on Enterococcus
faecalis, a Gram-positive coccus, and B. cereus, with
MIC values between 236 and 250 μg/mL. In fact, the
activities of compounds 14a–c against M. smegmatis are
approximately 10-fold higher than those of the
compared standard drug streptomycin (MIC: 0.46 μg/
mL). Although all the Mannich bases containing a β-
lactam nucleus (13a–c and 14a–c) exhibited inhibition
activity on the same microorganisms, the MIC values of
cephalosporin derivatives (14a–c) were better than those
of penicillin compounds (13a–c). Other Mannich bases
1H-1,2,4-triazol skeleton via
a methylene linkage,
exhibited the best antilipase activity with inhibitory rates
of 95% and 81% at a concentration of 10 μM,
respectively. Orlistat, a known pancreatic lipase inhibitor
used as an anti-obesity drug, exhibited an inhibitory
effect of 99% at
a
concentration of 300nM
(IC = 0.85nM). The IC values of compounds 13a and
50
50
15a were calculated at 1.41 and 10.43 μM, respectively.
Compounds 13a and 15a are potential alternatives to
orlistat.
α-Glucosidase inhibition.
All compounds were
containing piperazine (15a,b and 16a,b) or
a
evaluated with regard to α-glucosidase inhibition.
Compounds 14a and 15a, which may be regarded as
β-lactam derivatives, exhibited inhibition at various
concentrations with inhibitory rates of 74% and 54% of
morpholine nucleus (16c) were found to be active on
some of the test microorganisms used in the present
article (Table 3).
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

S. Demirci, A. Mermer, G. Ak, F. Aksakal, N. Colak, A. Demirbas, F. A. Ayaz, and N. Demirbas Vol 000
Table 4
the compounds’ AC values explained 81.45% of total
variation, where PC1 accounts for 48.18% of the variance
and PC2 for 33.28%. PC1 separated DPPH and
CUPRAC from the other AC assay, FRAP. First, DPPH,
following CUPRAC having a positive loading along the
axis on PC1, was associated with higher AC of
compounds 12c, 12e, 12a, and 12d, which are
fluoroquinolone derivatives, and 15b and 15a, which are
thiomorpholinomethyl-4,5-dihydro-1H-1,2,4-triazole
Pancreatic lipase, α-glucosidase, and urease inhibitory effects of synthe-
sized compounds (at final concentrations of 10 μM for lipase, 300 μM for
α-glucosidase, and 250 μM for urease).
Inhibition %
No.
Lipase
α-Glucosidase
Urease
3
—
—
45
29
—
—
—
—
38
—
—
—
81
—
—
95
—
—
—
99
—
—
1
36
69
13
—
22
—
16
—
38
70
—
23
—
—
—
—
75
—
75
6
7
7
8
8
9
9
1
a
a
b
a
b
a
derivatives carrying a phenylpiperazine nucleus. FRAP
was associated more with AC of compound 8a, a 1,3,4-
thiadiazole compound, and less with compound 9b, a
—
—
—
—
—
—
—
8
—
—
74
9
1
,3-thiazole compound, which all have a positive loading
b
0a
on PC1. In contrast, the remaining 17 compounds did not
lead to a complete separation for AC values depending
on the AC assay types. They were situated on the
negative and positive axes on PC2, contributing more or
less equal numbers of compounds at the left lower and
upper plans of the principal component. From the biplot,
some patterns can be seen in the distribution of
antioxidant capacity values of the synthesized compounds
among the assays.
The analyzed results for comparing the MW irradiation
method (MIM) and conventional method (CM) by PCA are
shown in Figure 2B. The principal component (PC) of
compounds listed in Table 1 (2, 3, 4, 6a, 6b, 7a, 7b, 9a, 9b,
1
1a
12a
12d
13a
14a
14c
15a
15b
16a
16c
54
—
18
2
Orlistat
Acarbose
Thiourea
83
100
Orlistat (at a final concentration of 0.3 μM), acarbose (at a final concen-
tration of 300 μM), and thiourea (at a final concentration of 250 μM) were
used as standard inhibitors.
1
0a, and 10b) explained 65.52% of total variation, where
PC1 accounts for 37.09% of the variance and PC2 for
8.44%. PCA indicated that compounds 10b and 10a were
—
, no data.
2
mostly closely associated and correlated with the power-
MIM and temp-MIM (r= 0.780, P<0.05) situated at the
right-lower plan on PC1. The remaining 11 compounds
were associated sometimes on PC1 (yield-CM with 7a and
7b) and PC2 (yield-MIM with 3 and 4, time-CM with 6a
and 6b) but were not correlated significantly (Fig. 2B).
The PCA (Fig. 2C) compared both methods with some
1
00μM, respectively (Table 4). Acarbose, an α-
glucosidase inhibitor used as an antidiabetic drug,
exhibited an inhibitory effect of 83% at the same
concentration. No significant inhibitory effect was
detected for other compounds.
Urease inhibition.
The synthesized compounds were
assayed for their in vitro inhibitory activity against Jack
Bean urease. Thiourea, with an IC50 value of 29.91 μM,
was used as a standard inhibitor. Initially, all synthesized
catalysts (InCl and p-TsOH), and compounds listed in
3
Table 2 (11a, 11b, 12a, 12b, 12c, 12d, 12e, 13a, 13b,
13c, 14a, 14b, 14c, 15a, 15b, 16a, 16b, and 16c)
explained a total variation of 64.27%. The power-MIM
(r = 0.625, P< 0.05), yield-MIM (r= 0.544, P< 0.05),
and time-MIM at the right-upper plan on PC1 were most
closely significantly associated and correlated with
compound 15a and less significantly with compounds
16c, 16b, 14c, and 11b, following time-CM. In addition,
yield-CM, yield-CM-CIn, and yield-CM-CTs (r =0.843,
0.912, P< 0.05) situated at the right-lower plan on PC1
were strongly correlated and associated with compounds
12b and 12e and less with 15b, 12d, and 12c,
respectively. The remaining five compounds, 13c, 11a,
12a, 16a, and 13b, with negative loadings and the
compounds, 14a, 13a, and 14b, with positive loadings
were situated on PC2 (variance 25.35%). The latter was
associated and correlated with the CM with catalysts
compounds were screened at
a
250-μM final
concentration. Among the synthesized compounds,
compounds 15b and 16c, thiomorpholinomethyl-4,5-
dihydro-1H-1,2,4-triazole
derivatives
carrying
a
phenylpiperazine (15b) or morpholine nucleus (16c),
exhibited the best inhibitory effects against urease (75%
inhibition). The other compounds exhibited no significant
inhibition (Table 4).
Principal component analysis. The antioxidant capacity
(AC) data of the synthesized 24 novel compounds
assayed using 2,2-diphenyl-1-picrylhydrazyl (DPPH),
ferric reducing ability of plasma (FRAP), and cupric ion
reducing antioxidant capacity (CUPRAC) applied to
principal component analysis (PCA) are shown in
Figure 2A. The results are presented in Table 3. PCA of
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2016
Antioxidant Capacity, Biological Activity, and Molecular Docking Studies of
New Hybrid Compounds
Figure 2. Biplot of scores and loadings of antioxidant capacity (AC) data from DPPH, FRAP, and CUPRAC results for 24 synthesized novel compounds
A); comparison of microwave irradiation method (MIM) and conventional method (CM) with compounds listed in Table 1 (B); comparison of MIM, CM,
(
and CM with catalysts (CM-cts) and the compounds listed in Table 2 (C); and comparison of enzyme inhibition by the compounds listed in Table 4 (D).
Abbreviations: y, yield; t, time; cts, catalyst. [Color figure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]
(
t-CM-cts, Fig. 2C). Values of enzyme inhibition using
with α-glucosidase; and compounds 13a, 7a, 7b, and 15a
and orlistat grouping with lipase on PC2 (variance
31.80%) with a negative loading. The PCs showed that
the inhibition of the three enzymes of 19 novel
synthesized compounds were associated but were not
correlated significantly.
Molecular docking results. Molecular docking reveals a
sufficient number of possible conformations and
orientations for an inhibitor at the binding site of the
enzyme. The most important interactions between
enzyme and inhibitor that stabilize the tertiary structure
of the enzyme can be determined from these
conformations. Docking of compounds to the active site
lipase, α-glucosidase, and urease enzymes against the
compounds listed in Table 2 (3, 6a, 7a, 7b, 8a, 8b, 9a,
9
b, 10a, 11a, 11b, 12a, 12d, 13a, 14a, 14c, 15a, 15b,
6a, and 16c) and the standard compounds orlistat,
1
acarbose, and thiourea were applied to PCA (Fig. 2D),
explaining 81.99% of total variation. PCA confirmed that
the synthesized compounds were related with the
inhibition of the three enzymes in three groups
established for compounds 9a, 3, 6a, 11a, 15a, 16c, 12d,
and 8a and thiourea grouping with urease on PC1
(
1
variance 50.19%) with a positive loading; compounds
4a, 14c, 16a, 12a, 9b, and 8b and acarbose grouping
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

S. Demirci, A. Mermer, G. Ak, F. Aksakal, N. Colak, A. Demirbas, F. A. Ayaz, and N. Demirbas Vol 000
of lipase, α-glucosidase, and urease enzymes was
performed. For comparison, two active compounds and
one inactive compound (7a) for all target enzymes were
selected according to the experimental results on their
inhibitory effects.
The most energetically profitable poses of the active
compounds 13a (81% inhibition) and 15a (95% inhibition)
and inactive compound 7a (45% inhibition) in the active
site of pancreatic lipase are presented in Figure 3.
For the inactive compound 7a, weak π–π interactions
were observed with Phe215 (at 3.8Å) and Phe77 (at
formation with all compounds. In addition to hydrogen
bonds with water molecules, two hydrogen bonds are
observed between 7a and both His351 and Tyr158
residues at a distance of 3.04Å. In addition, there is an
aromatic π–π interaction with Phe178 and the phenyl ring
of 7a at 3.6 Å. The binding affinity value is À12.0 kcal/
mol for compound 7a. A very strong hydrogen bond
formation was observed with –NH in the dimethyl amine
group of 14a and the oxygen atom of Gln279 (at 1.5Å).
Another hydrogen bond interaction is obtained with
Glu411 at 3.9 Å. There is also an electrostatic interaction
with the negatively charged oxygen atom of Gln279 and
positively charged sulfur in the six-membered
heterocyclic ring of 14a (at 2.6 Å) and π–H interaction
with Pro312 (at 3.7 Å). Compound 14a gives the highest
binding affinity value equal to À21.0kcal/mol with all
these interactions. A moderate hydrogen bond between
4
.5Å) residues with a binding affinity of À11.5 kcal/mol.
The energy of binding with lipase was considerably
higher for the active compounds 13a and 15a than for the
inactive compound 7a, at À23.2 and À23.5 kcal/mol,
respectively. Both compounds 13a and 15a make a long-
range hydrogen bond with His151 (at 4.3Å) residue.
Compound 13a has π–H interactions with Ile78 (at 3.6 Å)
and Arg256 (at 3.7Å) residues.
the –CH group of Arg315 and the sulfur in the six-
2
membered heterocyclic ring (at 3.07Å) and the π–H
interaction between the His280 and the five-membered
heterocyclic ring (at 4.3 Å) are obtained for compound
15a with the high affinity value of À18.7 kcal/mol.
The binding modes of the active compounds 14a (74%
inhibition) and 15a (54% inhibition) and the inactive
compound 7a (1% inhibition) to S. cerevisiae α-
glucosidase are shown in Figure 4.
As shown in Figure 4, the presence of water molecules
in the active site of the enzyme causes hydrogen bond
In Figure 5, inactive 7a (13% inhibition) and active 15b
(75% inhibition) and 16c (75% inhibition) are given as
examples of interaction with Helicobacter pylori urease.
Figure 3. 3D representation of docking poses 7a (a), 13a (b), and 15a (c) in the active site of pancreatic lipase (PDB code: 1LPB). [Color figure can be
viewed in the online issue, which is available at wileyonlinelibrary.com.]
Figure 4. 3D representation of docking poses 7a (a), 14a (b), and 15a (c) in the active site of Saccharomyces cerevisiae α-glucosidase (PDB code: 3A4A).
[Color figure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2016
Antioxidant Capacity, Biological Activity, and Molecular Docking Studies of
New Hybrid Compounds
Figure 5. 3D representation of docking poses 7a (a), 15b (b), and 16c (c) in the active site of Helicobacter pylori urease (PDB code: 1E9Y). [Color figure
can be viewed in the online issue, which is available at wileyonlinelibrary.com.]
Two hydrogen bonds formation with Cys321 (at 4.3 Å)
and Gly279 (at 3.8Å) residues and π–H interaction with
Ala169 (at 4.3 Å) are characteristics of the inactive
molecule 7a (Fig. 5a) with a low binding affinity value
equal to À8.0 kcal/mol. For compound 15b, two
hydrogen bonds with Gly280 (at 3.9Å) and Ala335 (at
anti-urease activity, 16c, incorporates a morpholine
nucleus linked to thiomorpholinomethyl-4,5-dihydro-1H-
1,2,4-triazol scaffold via a methylene linkage. Docking
some of the synthesized compounds into the active sites of
the lipase, α-glucosidase, and urease was also performed.
The results of the molecular docking revealed high binding
energy values (between À14.1 and À23.5kcal/mol) for
active compounds, while inactive compounds have lower
binding energy values (between À8 and À12kcal/mol).
4
.1Å) and π–H interaction with His322 (at 3.9 Å) are
obtained. Compound 16c is hydrogen bonded to Arg338
residue at a distance of 3.8Å. Binding affinity values for
1
5b and 16c are À15.0 and À14.1kcal/mol, respectively.
Two-dimensional representation of all these interactions
with corresponding enzymes is given as Supporting
Information (Figure S1).
EXPERIMENTAL
General. All the chemicals were purchased from Fluka
Chemie AG, Buchs, Switzerland, and used without further
purification. Melting points of the synthesized compounds
were determined in open capillaries on a Büchi B-540 melt-
ing point apparatus (Buchi, New Castle, DE) and are uncor-
rected. Reactions were monitored by TLC on silica-gel
60F254 aluminum sheets. The mobile phase was
ethanol/ethyl ether 1:1, and detection was performed using
UV light. FTIR spectra were recorded using a Perkin Elmer
1600 series FTIR spectrometer (Varian, Palo Alto, CA).
CONCLUSIONS
This article reports the conventional and eco-friendly
MW-irradiated synthesis of some new hybrid molecules
containing several heterocyclic units. The effect of acid
catalyst on the Mannich reaction was also investigated.
Antimicrobial, anti-urease, antilipase, anti-α-glucosidase,
and antioxidant activity screening studies were also
performed. Most of the newly synthesized compounds
exhibited good to moderate activities on some of the test
microorganisms. Of these, the compounds containing a
fluoroquinolone unit linked to the thiomorpholinomethyl-
1
13
H-NMR and
C-NMR spectra were registered in
DMSO-d on a BRUKER AVENE II 400-MHz NMR
6
1
spectrometer (Danbury, CT) (400.13MHz for H and
100.62MHz for C) or Varian-Mercury 200MHz NMR
spectrometer (Varian, Palo Alto, CA) (200 MHz for H
and 50MHz for C). MW-assisted syntheses were carried
out using a monomode CEM-Discover MW apparatus
(CEM Corporation, Matthews, NC). The chemical shifts
1
3
1
4
,5-dihydro-1H-1,2,4-triazol scaffold via a methylene
13
linkage exhibited excellent activity on the test bacteria.
Moreover, all compounds 12 (except 12b) demonstrated
radical scavenging activity. Compound 15a containing a
methoxyphenylpiperazine nucleus exhibited enzyme
inhibition on lipase and α-glucosidase, while 15b with no
methoxy group on the phenyl ring displayed enzyme
inhibition on urease, in addition to its radical scavenging
activity. Compounds 13a and 14a, which are hybrid
compounds with a β-lactam unit, were found to exhibit
inhibitory activity on the enzymes lipase and α-
glucosidase, respectively. The other compound displaying
are given in parts per million relative to Me Si as an internal
4
reference; j values are given in hertz. The elemental analy-
sis was performed on a Costech Elemental Combustion
System CHNS-O elemental analyzer (Costech Analytical
Technologies, Inc., Italy). All the compounds elicited C,
H, and N analysis within ±0.4% of the theoretical values.
The mass spectra were obtained on a Quattro LCMS
(70 eV) instrument (Waters, Micromass, Manchester, UK).
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

S. Demirci, A. Mermer, G. Ak, F. Aksakal, N. Colak, A. Demirbas, F. A. Ayaz, and N. Demirbas Vol 000
Ethyl 2-thiomorpholinoacetate (2). In method 1, ethyl
bromoacetate (10 mmol) was added to a mixture of
thiomorpholine (10 mmol) and triethylamine (10 mmol)
dropwise in dry tetrahydrofuran at 0–5°C. The reaction
content was allowed to reach 60°C and was stirred for
refluxed for 15h in the presence of CS (1.52 g, 20mmol).
2
It was then cooled to room temperature and acidified to
pH 6 with 37% HCl. The mixture was cooled overnight,
producing a solid. This was recrystallized from ethyl
acetate to give the target compound. Yield: 53%, mp
203–204°C. In method 2, the solution of KOH (1 mmol)
in water was added to the solution of compound 3 in
5 mL of water and 5 mL of ethanol. The mixture was
irradiated in monomode MW reactor in closed vessel
with the pressure control at 150 W for 8 min (hold time).
It was then cooled to room temperature and acidified to
pH 6 with 37% HCl. The mixture was cooled overnight,
producing a solid. This was recrystallized from ethyl
acetate to give the target compound. Yield: 93%, mp
2
4h; the progress of the reaction was monitored by TLC.
The precipitated triethylammonium salt was removed by
filtration. After the solvent was evaporated under reduced
pressure, a yellow solid appeared. This crude product was
recrystallized from ethanol and ethyl acetate (2:1) to
afford the desired product. In method 2, ethyl
bromoacetate (10 mmol) was added to a mixture of
thiomorpholine (10 mmol) and triethylamine (10 mmol)
dropwise in dry tetrahydrofuran at 0–5°C for 5min and
then irradiated in closed vessels with the pressure control
at 110°C for 15min (hold time) at 200-W maximum
power. The precipitate was removed by filtration, and the
resulting solution was evaporated under reduced pressure
to dryness. The yellow solid was recrystallized from
ethanol and ethyl acetate (2:1) to afford the desired
À1
203–204°C. FTIR (υmax, cm ): 3313 (NH), 1540 (C=N),
1
1286 (C=S). H NMR (DMSO-d , δ ppm): 2.50 (s, 4H,
6
13
2CH ), 2.60 (s, 4H, 2CH ), 3.70 (s, 2H, CH ). C NMR
2
2
2
(DMSO-d , δ ppm): 27.49 (2CH ), 52.47 (CH ), 54.17
6
2
2
(2CH ), 161.22 (oxadiazole C-5), 178.46 (oxadiazole C-
2
+
2). LC MS m/z (%):216.21 ([M-1] 100), 238.09 ([M-2
À1
+
product. Yield: 88%, mp 80°C. FTIR (υmax, cm ): 1732
+ Na] 10), 159.07 (10), 116.15 (25). Elemental analysis
1
(
C=O), 1243 (C-O). H NMR (DMSO-d , δ ppm): 1.18
for C H N OS . Calculated (%): C: 38.69; H: 5.10, N:
6
7
11
3
2
(
t, 3H, CH , J= 7.2Hz), 2.56 (t, 4H, 2CH , J= 4.8 Hz),
19.34. Found (%): C: 38.42; H: 5.29; N: 19.61.
3
2
2
2
1
6
.76 (t, 4H, 2CH , J= 4.8Hz), 3.25 (s, 2H, CH ), 4.07 (q,
General method for the synthesis of compounds (5a–e).
A
2
2
1
3
H, OCH , J = 7.2Hz). C NMR (DMSO-d , δ ppm):
solution of compound 3 (10 mmol) in absolute ethanol
was refluxed with benzaldehyde (for 5a), 4-
methoxibenzaldehyde (for 5b), 4-pyridinecarboxaldehyde
(for 5c), 3-hydroxy-4-methoxy benzaldehyde (for 5d), and
indol-3-carbaldehyde (for 5e) (10 mmol) for 6 h. The
reaction content was allowed to reach room temperature,
at which a solid appeared. This crude product was filtered
off and recrystallized from acetone to obtain the desired
compound.
2
6
4.60 (CH ), 27.66 (2CH ), 47.60 (CH ), 53.97 (2CH ),
3
2
2
2
7.48 (OCH ), 170.57 (C=O) [30].
2
2
-Thiomorpholinoacetohydrazide (3).
In method 1,
hydrazine hydrate (0.60 mL, 25 mmol) was added to a
solution of compound 2 (10 mmol) in absolute ethanol,
and the mixture was allowed to reflux for 12h. When the
reaction mixture was cooled to room temperature, a white
solid appeared. The crude product was filtered off and
recrystallized from ethanol to give the desired compound.
Yield: 50%, mp 102–103°C. In method 2, the solution of
compound 2 (1mmol) in hydrazine hydrate (2.5 mmol)
was irradiated in monomode MW reactor in closed vessel
with the pressure control at 100W for 7min (hold time).
After adding ethanol, a white solid appeared. This crude
product was filtered off and recrystallized from ethanol.
N′-Phenylmethylene-2-thiomorpholin-4-ylaceto
hydrazide
À1
1
(5a). Yield: 90%, mp 157–158°C. FTIR (υmax, cm ):
3265 (NH), 2927 (ar-CH), 1770 (C=O), 1449 (C=N). H
NMR (DMSO-d , δ ppm): 2.58–2.86 (m, 8H, 4CH ),
6
2
3.10 (s, 2H, CH ), 7.38–7.41 (m, 3H, ar-H), 7.65–7.67
2
(m, 2H, ar-H), 7.94 and 8.33 (1H, s, N=CH, cis/trans
conformers), 11.11 and 11.29 (1H, s, NH, cis/trans
À1
13
Yield: 97%, mp 102–103°C. FTIR (υmax, cm ): 3316,
3
conformers).
C NMR (DMSO-d , δ ppm): 27.38
6
1
289 (NH ), 3212 (NH), 1644 (C=O). H NMR (DMSO-
(2CH ), 54.98 (2CH ), 61.56 (CH ), arC: [127.14 (2CH),
2 2 2
2
d , δ ppm): 2.51 (bs, 4H, 2CH + DMSO-d ), 2.93 (s, 2H,
127.46 (2CH), 129.23 (CH), 134.71 (C)], 147.72
6
2
6
CH ), 3.40 (bs, 4H, 2CH ), 4.22 (bs, 2H, NH , D O
(N=CH), 171.26 (C=O). LC MS m/z (%): 264.40 ([M
+ 1] 100), 116.09 (80). Elemental analysis for
2
2
2
2
1
3
+
exch.), 8.91 (s, H, NH, D O exch.). C NMR (DMSO-
2
d , δ ppm): 27.48 (2CH ), 55.00 (2CH ), 61.04 (CH ),
C H N OS. Calculated (%): C: 59.29; H: 6.51, N:
13 17 3
6
2
2
2
1
+
68.72 (C=O). LC MS m/z (%): 216.21 ([M+ K
2H] 100), 159.07 (38), 142.05 (65). Elemental analysis
15.96. Found (%): C: 59.22; H: 6.59; N: 15.61.
+
N′-[(4-Methoxyphenyl)methylene]-2-thiomorpholin-4-ylace-
for C H N OS. Calculated (%): C: 41.12; H: 7.48; N:
tohydrazide (5b).
Yield: 93%, mp 131–132°C. FTIR
6
13 3
À1
2
3.98%. Found (%): C: 41.16; H: 7.49; N: 23.91.
-(Thiomorpholinomethyl)-1,3,4-oxadiazole-2-thiol (4). In
(υmax, cm ): 3263 (NH), 3019 (ar-CH), 1766 (C=O),
1
5
1455 (C=N). H NMR (DMSO-d , δ ppm): 2.58–2.85
6
method 1, the solution of KOH (0.56g, 10 mmol) in water
(m, 8H, 4CH ), 3.10 (s, 2H, CH ), 3.80 (s, 3H, OCH ,),
2
2
3
was added to the solution of compound 3 in water (50 mL,
6. 93–6.99 (m, 3H, ar-H), 7.56–7.65 (m, 2H, ar-H), 7.92
and 8.22 (1H, s, N=CH, cis/trans conformers), 10.95 and
1
0mmol) and ethanol (50 mL, 10mmol). The mixture was
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2016
Antioxidant Capacity, Biological Activity, and Molecular Docking Studies of
New Hybrid Compounds
1
3
1
(
(
1
1
1
1.08 (1H, s, NH, cis/trans conformers). C NMR
General method for the synthesis of compounds 6a and
6b. In method 1, a mixture of compound 3 (10 mmol)
and benzylisothiocyanate (for 6a) or phenylisocyanate
(for 6b) in absolute ethanol was refluxed for 18h. When
the reaction content was cooled to room temperature, a
white solid formed. This crude product was filtered off
and recrystallized from ethyl acetate :hexane (1:2) to
afford the desired compound. In method 2, a mixture of
compound 3 (1 mmol) and benzylisothiocyanate (for 6a)
or phenylisocyanate (for 6b) (1mmol) in absolute ethanol
was irradiated in monomode MW reactor in closed vessel
with the pressure control at 120 W for 7 min (hold time).
The solid obtained following the addition of water was
filtered off and recrystallized from ethyl acetate :hexane
(1:2) to afford the desired compound.
DMSO-d , δ ppm): 27.72 (2CH ), 54.99 (2CH ), 55.71
OCH ), 61.59 (CH ), arC: [114.73 (2CH), 127.25 (C),
28.69 (2CH)], 147.60 (N=CH), 160.99 (C-OCH ),
71.05 (C=O). LC MS m/z (%): 294.40 ([M +1] 100),
6
2
2
3 2
3
+
16.29 (94). Elemental analysis for C H N O .
1
4 19 3 2
Calculated (%): C: 57.31; H: 6.53, N: 14.32. Found (%):
C: 57.32; H: 6.59; N: 14.31.
N′-[Pyridin-4-ylmethylene]-2-thiomorpholin-4-ylacetohydra-
zide (5c). Yield: 95%, mp 78–79°C. FTIR (υmax, cm ):
264 (NH), 3070 (ar-CH), 1689 (C=O), 1405 (C=N). H
NMR (DMSO-d , δ ppm): 2.58–2.85 (m, 8H, 4CH ),
.50 (s, 2H, CH +H O), 7.53–7.59 (m, 3H, ar-H), 7.92
and 8.32 (1H, s, N=CH, cis/trans conformers), 8.56–8.59
m, 2H, ar-H), 11.35 and 11.68 (1H, s, NH, cis/trans
C NMR (DMSO-d , δ ppm): 27.62
2CH ), 54.85 (2CH ), 61.51 (CH ), arC: [121.28 (2CH),
41.91 (C), 145.35 (2CH)], 150.49 (N=CH), 171.62
C=O). LC MS m/z (%): 265.12 ([M + 1] 20), 164.12
À1
1
3
6
2
3
2 2
(
13
conformers).
(
1
(
(
N-Benzyl-2-(2-thiomorpholinoacetyl)hydrazinecarbothio-
amide (6a). Yield: 62% (method 1), 98% (method 2); mp
6
2 2 2
À1
161–162°C. FTIR (υmax, cm ): 3229 (3NH), 3088 (ar-
+
1
CH), 1673 (C=O), 1288 (C=S). H NMR (DMSO-d , δ
6
+
[M] 20), 116.96 (100). Elemental analysis for
ppm): 2.62 (bs, 4H, 2CH ), 2.70 (bs, 4H, 2CH ), 3.04 (s,
2
2
C H N OS. Calculated (%): C: 54.52; H: 6.10, N:
2H, CH ), 4.72 (d, 2H, CH , J= 8.0 Hz), 7.20–7.32 (m,
12
16
4
2 2
2
1.19. Found (%): C: 54.32; H: 6.19; N: 21.11.
N′-[(3-Hydroxy-4-methoxyphenyl)methylene]-2-thiomorpho-
5H, arH), 8.36 (bs, 1H, NH, D O exch.), 9.30 (bs, 1H,
2
13
NH, D O exch.), 9.70 (bs, 1H, NH, D O exch.).
C
2
2
lin-4-ylacetohydrazide (5d). Yield: 95%, mp 104–106°C.
NMR (DMSO-d , δ ppm): 27.42 (2CH ), 47.17 (2CH ),
54.98 (CH ), 61.01 (CH ), arC: [127.08 (2CH), 127.48
2 2
6 2 2
À1
FTIR (υmax, cm ): 3564 (OH), 3237 (NH), 2917 (ar-
1
CH), 1651 (C=O), 1435 (C=N). H NMR (DMSO-d , δ
(CH), 128.51 (2CH), 139.69 (C)], 169.01 (C=O), 182.04
6
+
ppm): 2.58–2.70 (m, 8H, 4CH ), 3.06 (s, 2H, CH ), 3.77
(C=S). LC MS m/z: 363.21 ([M + K] 10), 347.30 ([M
2
2
+
+
+
(
s, 3H, OCH ) 6.92–6.94 (m, 2H, ar-H), 7.19 (s, 1H, ar-
+ Na] 15), 326.23 ([M +2] 10), 325.28 ([M + 1] 57).
3
H), 7.82 and 8.12 (1H, s, N=CH, cis/trans conformers),
.35 (bs, 1H, OH, D O exch.), 10.95 and 11.08 (1H, s,
Elemental analysis for C H N OS . Calculated (%): C:
51.82; H: 6.21; N: 17.27%. Found (%): C: 51.86; H:
6.29; N: 17.30.
14
20
4
2
9
2
1
3
NH, cis/trans conformers). C NMR (DMSO-d , δ
6
ppm): 27.39 (2CH ), 55.25 (2CH ), 55.81 (OCH ), 61.34
N-Phenyl-2-(2-thiomorpholinoacetyl)hydrazine carboxamide
(6b). Yield 65% (method 1), 85% (method 2); mp 165°C.
2
2
3
(
CH ), arC: [114.62 (CH), 127.19 (C), 128.69 (2CH),
2
À1
1
1
1
47.80 (N=CH), 161.37 (C-OH), 165.80 (C-OCH )],
71.03 (C=O). LC MS m/z (%): 310.50 ([M + 1] 75),
FTIR (υmax, cm ): 3298, 3195 (3NH), 3042 (ar-CH),
3
+
1
1703 (C=O), 1665 (C=O). H NMR (DMSO-d , δ ppm):
6
16.35 (100). Elemental analysis for C H N O S.
2.65 (bs, 4H, 2CH ), 2.71 (bs, 4H, 2CH ), 3.05 (s, 2H,
1
4
19
3
3
2
2
Calculated (%): C: 54.35; H: 6.19, N: 13.58. Found (%):
C: 54.32; H: 6.19; N: 13.51.
N′-[1H-Indol-2-ylmethylene]-2-thiomorpholin-4-ylacetohy-
CH ), 7.15–7.30 (m, 5H, arH), 8.30 (bs, 1H, NH, D O
2 2
exch.), 9.34 (bs, 1H, NH, D O exch.), 9.72 (bs, 1H, NH,
2
13
D O exch.).
2
C NMR (DMSO-d , δ ppm): 27.38
6
drazide (5e). Yield: 91%, mp 194–195°C. FTIR (υmax
,
(2CH ), 53.83 (2CH ), 54.17 (CH ), arC: [127.79 (2CH),
2 2 2
À1
cm ): 3214 (NH), 2968 (ar-CH), 1766 (C=O), 1435
128.58 (CH), 129.57 (2CH), 134.28 (C)], 152.44 (C=O),
1
+
(
C=N). H NMR (DMSO-d , δ ppm): 2.48–2.75 (m, 8H,
171.24 (C=O). LC MS m/z: 296.23 ([M+ 2] 30), 295.28
6
+
4
CH ), 3.08 (s, 2H, CH ), 7.16 (m, 1H, ar-H, J= 8.0 Hz),
([M + 1] 78), 123.27 (100). Elemental analysis for
2
2
7
.42 (d, 1H, ar-H, J =8.0 Hz), 7.74 (bs, 1H, ar-H), 8.08
C H N O S. Calculated (%): C: 53.04; H: 6.16; N:
1
3 18 4 2
(d, 1H, ar-H, J= 8.0Hz), 8.19 (d, 1H, ar-H, J= 8.0 Hz),
19.03. Found (%): C: 53.06; H: 6.19; N: 19.01.
8
.13 and 8.46 (1H, s, N=CH, cis/trans conformers),
General method for the synthesis of compounds 7a and
7b. In method 1, a solution of compounds 6a and 6b
(10 mmol) in ethanol :water (1:1) was refluxed in the
presence of 2 N of NaOH for 3 h, and then, the resulting
solution was cooled to room temperature and acidified to
pH 7 with 37% HCl. The precipitate that formed was
filtered off, washed with water, and recrystallized from
ethanol :water (1:1) to afford the desired compound. In
method 2, the solution of 2N of NaOH (2.5 mL) was
13
1
0.77 and 10.98 (1H, s, NH, cis/trans conformers).
C
NMR (DMSO-d , δ ppm): 27.45 (2CH ), 55.34 (2CH ),
6
6
2
2
1.70 (CH ), arC: [112.23 (CH), 120.74 (CH), 121.91
2
(
(
CH), 123.12 (CH), 124.51 (C), 130.70 (CH), 138.00
2C), 144.74 (N=CH), 165.27 (C=O). LC MS m/z (%):
+
3
03.33 ([M + 1] 100), 116.10 (90). Elemental analysis
for C H N O S. Calculated (%): C: 59.58; H: 6.00, N:
1
14 19 3 3
8.53 Found (%): C: 59.59; H: 6.00; N: 18.53.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

S. Demirci, A. Mermer, G. Ak, F. Aksakal, N. Colak, A. Demirbas, F. A. Ayaz, and N. Demirbas Vol 000
added to the solution of corresponding compound 6
1 mmol) in ethanol : water (1:1), and the mixture was
(DMSO-d , δ ppm): 27.50 (2CH ), 54.28 (CH ), 54.67
6 2 2
(
(2CH ), 61.39 (CH ), 127.08 (CH), 127.17 (CH), 127.40
2 2
stirred at room temperature for 5 min. Then, the mixture
was irradiated in monomode MW reactor in closed vessel
with the pressure control at 200W for 5–7min (hold
time). Upon acidification of reaction content to pH7 with
(CH), 127.81 (CH), 127.94 (CH), 141.21 (C), 160.56
(thiadizole C-2), 176.40 (thiadiazole C-5). LC MS m/z:
+
+
345.28 ([M + K] 39), 324.28 ([M + H O] 15), 323.40
2
+
+
([M-1 + H O] 100), 305.35 ([M-1] 10). Elemental
2
3
7% HCl, a white solid appeared. This crude product was
analysis for C H N S . Calculated (%): C: 54.87; H:
1
4 18 4 2
filtered off, washed with water, and recrystallized from
5.92; N: 18.28. Found (%): C: 54.89; H: 5.95; N: 185.23.
ethanol : water (1:1) to afford the desired compound.
N-Phenyl-5-(thiomorpholinomethyl)-1,3,4-oxadiazol-2-
À1
4
-Benzyl-5-(thiomorpholinomethyl)-4H-1,2,4-triazole-3-thiol
amine (8b). Yield: 90%, mp 155°C. FTIR (υmax, cm ):
1
(
1
(
7a). Yield: 75% (method 1), 98% (method 2); mp 194–
3519 (NH), 3090 (ar-CH). H NMR (DMSO-d , δ ppm):
6
À1
95°C. FTIR (υmax, cm ): 3087, 3033 (ar-CH), 2831
2.68 (bs, 4H, 2CH ), 2.76 (bs, 4H, 2CH ), 3.37 (bs, 2H,
2
2
1
SH). H NMR (DMSO-d , δ ppm): 2.31 (s, 4H, 2CH ),
CH ), 7.95 (d, 2H, arH, J = 6.1 Hz), 7.26 (t, 1H, arH,
6
2
2
2
.50 (s, 2H, CH + DMSO-d ), 3.41 (s, 4H, 2CH ), 5.29
J = 4.8 Hz), 7.41 (d, 2H, arH, J= 6.3 Hz), 8.71 (bs, 1H,
2
6
2
13
(s, 2H, CH ), 7.20 (d, 2H, arH, J= 7.2Hz), 7.28 ( d, 1H,
NH, D O exch.). C NMR (DMSO-d , δ ppm): 27.50
2
2
6
arH, J = 6.8Hz), 7.34 (t, 2H, arH, J = 7.2Hz), 13.80 (bs,
(2CH ), 54.28 (CH ), 54.69 (2CH ), arC: [127.08 (2CH),
2 2 2
127.10 (CH), 127.21 (2CH), 141.24 (C)], 160.55
1
3
1
H, SH, D O exch.). C NMR (DMSO-d , δ ppm):
2
6
2
7.12 (2 CH ), 46.69 (CH ), 53.20 (CH ), 54.45 (2 CH ),
(thiazole C-2), 176.42 (thiazole C-5). LC MS m/z: 275.35
([M-1] 35), 198.20 (100). Elemental analysis for
2
2
2
2
+
arC: [127.24 (2CH), 127.78 (CH), 128.86 (2CH), 136.61
C)], 149.58 (triazole C-5), 168.73 (triazole C-3). LC MS
(
C H N OS. Calculated (%): C: 56.50; H: 5.84; N:
13 16 4
+
+
m/z: 329.31 ([M + Na] 10), 323.27 ([M-1 +H O] 15),
20.27. Found (%): C: 56.51; H: 5.85; N: 20.23.
2
+
+
3
08.24 ([M+ 2] 25), 307.30 ([M+ 1] 100). Elemental
General method for the synthesis of compounds 9a and
analysis for C H N S . Calculated (%): C: 54.87; H:
9b.
In method 1, 4-chlorophenacylbromide (10 mmol)
1
4 18 4 2
5
.92; N: 18.28. Found (%): C: 54.86; H: 5.99; N: 18.31.
and dried sodium acetate (200mmol) were added to a
solution of compounds 6a and 6b in absolute ethanol,
and the reaction mixture was refluxed for 18h. The
mixture was cooled to room temperature, poured into ice-
cold water while stirring, and left overnight in the cold.
The formed solid was filtered, washed with water three
times, and recrystallized from ethyl acetate : n-hexane
(1:1) to afford the desired compound. In method 2, 4-
chlorophenacylbromide (1 mmol) and dried sodium
acetate (20 mmol) were added to a solution of the
corresponding compound 6 in absolute ethanol, and the
reaction mixture was irradiated in monomode MW
reactor in closed vessel with the pressure control at
200 W for 30min (hold time). The mixture was cooled to
room temperature, poured into ice-cold water while
stirring, and left overnight in the cold. The formed solid
was filtered, washed with water three times, and
recrystallized from ethyl acetate : n-hexane (1:1) to afford
the desired compound.
4
-Phenyl-5-(thiomorpholinomethyl)-4H-1,2,4-triazol-3-ol
(
7b). Yield: 82% (method 1), 88% (method 2); mp 191–
À1
1
1
92°C. FTIR (υmax, cm ): 3280 (OH), 3058 (ar-CH). H
NMR (DMSO-d , δ ppm): 2.36 (s, 4H, 2CH ), 2.48 (s,
6
2
4
H, 2CH ), 3.30 (s, 2H, CH ), 6.93 (s, 1H, NH, D O
2 2 2
exch.), 7.45–7.48 (m, 5H, arH), 7.96 (s, 1H, NH, D O
exch.), 8.79 93 (s, 1H, NH, D O exch.), 13.51 (s, 1H,
OH, D O exch.). C NMR (DMSO-d , δ ppm): 27.38
2
2
1
3
2
6
(2CH ), 53.83 (CH ), 54.16 (2CH ), arC: [127.19 (2CH),
2 2 2
1
28.85 (CH), 129.53 (2CH), 134.21 (C)], 144.52 (triazole
+
C-5), 153.24 (triazole C-3). LC MS m/z: 278.24 ([M + 2]
5), 197.30 (100). Elemental analysis for C H N OS.
5
1
3 16 4
Calculated (%): C: 56.50; H: 5.84; N: 20.27. Found (%):
C: 56.56; H: 5.89; N: 20.23.
General method for the synthesis of compounds 8a and
8
b.
Concentrated sulfuric acid (28 mL, 64 mmol) was
added to compounds 6a and 6b (10 mmol) dropwise
while stirring. The reaction content was stirred in an ice
bath for 15min. The mixture was allowed to reach room
temperature and was then stirred for an additional 2h.
The resulting solution was then poured into ice-cold
water and made alkaline (pH 8) with ammonia. The
precipitated product was filtered, washed with water, and
recrystallized from methanol to afford the desired product.
N-Benzyl-5-(thiomorpholinomethyl)-1,3,4-thiadiazol-2-
N′-(3-Benzyl-5-(4-chlorophenyl)thiazol-2(3H)-ylidene)-2-
thiomorpholinoacetohydrazide (9a).
1), 89% (method 2); mp 119–120°C. FTIR (υ , cm ):
3214 (NH), 3058, 3024 (ar-CH), 1725 (C=O). H NMR
Yield: 40% (method
À1
max
1
(DMSO-d , δ ppm): 2.51 (s, 4H, 2CH ), 2.59 (s, 4H,
6
2
2CH ), 3.60 (s, 2H, CH ), 4.36 (s, 2H, CH ), 7.25–7.27
2
2
2
(m, 2H, 2CH), 7.28–7.31 (m, 6H, arH), 8.12 (t, 2H, 2CH,
13
amine (8a).
Yield: 97%, mp 70–71°C. FTIR (υmax,
J = 4.0 Hz). C NMR (DMSO-d , δ ppm): 27.54 (2CH ),
6 2
À1
1
cm ): 3067 (NH), 3033 (ar-CH), 1287 (C-S). H NMR
46.49 (CH ), 52.53 (CH ), 54.28 (2 CH ), arC: [127.58
2 2 2
(
2
(
DMSO-d , δ ppm): 2.65 (bs, 4H, 2CH ), 2.66 (bs, 4H,
(2CH+ thiazole C-5), 127.81 (3CH), 128.80 (3CH),
129.90 (CH), 139.25 (2C+ thiazole C-4)], 157.16
(thiazole C-2), 164.35 (C=O). LC MS m/z: 478.35 ([M
6
2
CH ), 2.90 (bs, 2H, CH ), 4.23 (s, 2H, CH ), 7.26–7.38
2 2 2
13
m, 5H, arH), 9.68 (bs, 1H, NH, D O exch.). C NMR
2
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2016
Antioxidant Capacity, Biological Activity, and Molecular Docking Studies of
New Hybrid Compounds
+
+
+
1 +H O] 10), 455.39 (18), 443.31 ([M +2-H O] 15),
C H N O S . Calculated (%): C: 52.72; H: 5.53; N:
16 20 4 2 2
2
2
3
65.29 (36), 153.12 (100). Elemental analysis for
15.37. Found (%): C: 52.59; H: 5.45; N: 15.31.
C H ClN OS . Calculated (%): C: 57.56; H: 5.05; N:
N′-(4-Oxo-3-phenyloxazolidin-2-ylidene)-2-thiomorpholino-
22
23
4
2
1
2.21 Found (%): C: 57.55; H: 5.05; N: 12.28.
N′-(5-(4-Chlorophenyl)-3-phenyloxazol-2(3H)-ylidene)-2-
acetohydrazide (10b).
(method 2); mp 174°C. FTIR (υmax, cm ): 3515 (NH),
3093 (ar-CH), 1798 (2C=O). H NMR (DMSO-d , δ
ppm): 2.59 (s, 4H, 2CH ), 2.65 (s, 4H, 2CH ), 3.14 (s,
Yield: 50% (method 1), 92%
À1
1
thiomorpholinoacetohydrazide (9b).
Yield: 70% (method
6
À1
1
3
), 97% (method 2); mp 210–211°C. FTIR (υmax, cm ):
215 (NH), 3023 (ar-CH), 1715 (C=O). H NMR
2
2
1
2H, CH ), 4.84 (s, 2H, CH ), 6.90 (bs, 1H, arH), 7.22
2 2
(
2
DMSO-d , δ ppm): 2.66 (bs, 4H, 2CH ), 2.74 (bs, 4H,
(bs, 1H, arH), 7.49 (bs, 1H, arH), 9.08 (bs, 1H, NH, D O
2
6
2
13
CH ), 3.05 (s, 2H, CH ), 6.92 (s, 1H, arH), 6.96 (d, 2H,
exch.). C NMR (DMSO-d , δ ppm): 27.70 (2CH ),
2
2
6 2
arH, J = 7.1Hz), 7.24 (t, H, arH, J= 5.8Hz), 7.41 (d, 2H,
45.93 (CH ), 53.14 (2CH ), 61.27 (thiazole C-5), arC:
2 2
1
3
arH, J = 7.3Hz), 7.54 (m, 4H, arH). C NMR (DMSO-
d , δ ppm): 27.53 (2CH ), 46.49 (CH ), 54.23 (2 CH ),
[127.94 (CH), 128.28 (2CH), 129.85 (2CH), 136.54 (C)],
158.84 (thiazole C-2), 165.64 (C=O), 171.92 (thiazole C-
6
2
2
2
+
+
arC: [120.09 (oxazole C-4), 122.58 (2CH), 124.81
4). LC MS m/z: 336.35 ([M + 2] 15), 334.29 ([M] 47),
(2CH), 125.80 (2CH), 128.90 (2CH), 129.54 (CH),
126.31 (100). Elemental analysis for C H N O S.
1
5 18 4 3
1
1
37.65 (C), 139.35 (C + oxazole C-5), 145.52 (C)],
Calculated (%): C: 53.88; H: 5.43; N: 16.75. Found (%):
C: 53.89; H: 5.45; N: 16.71.
58.16 (oxazole C-2), 164.35 (C=O). LC MS m/z: 428.35
+
([M] 35), 251.12 (100). Elemental analysis for
General method for the synthesis of compounds 11a, 11b,
12a–e, 13a–c, 14a–c, 15a, 15b, and 16a–c. In method 1,
the appropriate secondary amine (10 mmol) was added
to a solution of compound 4 (10mmol) (for 11c, 12d,
12e, 13c, 14c, and 15b), compound 7a (10 mmol) (for
11a, 12a, 13a, 14a, and 16a–c), and compound 7b (for
11b, 12b, 12c, 13b, 14b, and 15a) (10 mmol) in dry
tetrahydrofuran. The mixture was stirred at room
temperature in the presence of formaldehyde (37%,
30mmol) for 3 h. The solvent was then evaporated
under reduced pressure, and a solid appeared. The crude
product was recrystallized from an appropriate solvent
to give the desired compound [for 11a and 16a; ethyl
acetate, for 11b, 12a–e, 13a–c, 14a–c, 15a, 15b;
DMSO : water (1:1), for 16b and 16c; n-butyl acetate :
diethyl ether (1:2)]. In method 2, the appropriate
secondary amine (1 mmol) was added into a solution of
compound 4 (1 mmol) (for 11c, 12d, 12e, 13c, 14c, and
15b), compound 7a (1mmol) (for 11a, 12a, 13a, 14a,
and 16a-c), or compound 7b (for 11b, 12b,c, 13b, 14b,
and 15a) (1 mmol) in dry tetrahydrofuran, and the
mixture was irradiated in monomode MW reactor in
closed vessel (physical parameters were presented in
Table 1). The resulting solution was then poured into
ice water. The product precipitated was filtered off and
recrystallized from an appropriate solvent to give the
desired compound. In method 3, the solution of suitable
amine (10 mmol) in dimethyl formamide was stirred at
room temperature in the presence of formaldehyde
(37%, 30mmol) for 15min. Compound 4 (10 mmol) (for
11c, 12d, 12e, 13c, 14c, and 15b), compound 7a
(10 mmol) (for 11a, 12a, 13a, 14a, and 16a-c), or
compound 7b (for 11b, 12b, 12c, 13b, 14b, and 15a)
(10 mmol) was added to it. Stirring was continued in the
presence of catalytic amount of a suitable catalyst as
listed in Table 2. The reaction mixture was poured into
ice water, and a solid was obtained. This crude product
C H ClN O S. Calculated (%): C: 58.80; H: 4.93; N:
21
21
4 2
1
3.06. Found (%): C: 58.85; H: 4.95; N: 13.08.
General method for the synthesis of compounds 10a and
1
0b.
In method 1, ethyl bromoacetate (10 mmol) was
added to the solution of the corresponding compound 6 in
absolute ethanol (10 mmol), and the mixture was refluxed
in the presence of dried sodium acetate (16.4 g, 20mmol)
for 12–18h. The mixture was then cooled to room
temperature, poured into ice-cold water while stirring, and
left overnight in the cold. The formed solid was filtered,
washed with water three times, and recrystallized from
ethyl acetate : n-hexane (1:1) to afford the pure desired
compound. In method 2, ethyl bromoacetate (1mmol)
was added to the solution of corresponding compound 6
in absolute ethanol (1 mmol), and the mixture was
irradiated in monomode MW reactor in closed vessel with
the pressure control at 150 W (for 10a) or 200W (for
1
0b) for 10–15 min (hold time) (Table 1). The mixture
was then cooled to room temperature, poured into
ice-cold water while stirring, and left overnight in the
cold. The formed solid was filtered, washed with water
three times, and recrystallized from ethyl acetate :
n-hexane (1:1) to afford the pure desired compound.
N′-(3-Benzyl-4-oxothiazolidin-2-ylidene)-2-thiomorpholino-
acetohydrazide (10a).
Yield: 75% (method 1), 90%
À1
(method 2); mp 174°C. FTIR (υmax, cm ): 3209 (NH),
1
3
032, 2979 (ar-CH), 1726, 1696 (2C=O). H NMR
(
DMSO-d , δ ppm): 2.50 (s, 4H, 2CH ), 2.65 (s, 4H,
6
2
2
CH ), 3.06 (s, 2, CH ), 4.14 (s, 2H, CH ), 4.84 (s, 2H,
2 2 2
CH ), 7.27–7.37 (m, 5H, arH), 10.08 (bs, 1H, NH, D O
2
2
13
exch.). C NMR (DMSO-d , δ ppm): 27.70 (2CH ),
6
2
3
3.14 (thiazole C-5), 45.93 (CH ), 54.94 (2CH ), 61.27
2 2
(
CH ), arC: [127.97 (CH), 128.23 (2CH), 128.85 (2CH),
2
1
36.54 (C)], 158.84 (thiazole C-2), 165.64 (C=O), 171.92
+
(
(
thiazole C-4). LC MS m/z: 366.35 ([M + 2] 10), 365.29
[M + 1] 47), 116.21 (100). Elemental analysis for
+
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

S. Demirci, A. Mermer, G. Ak, F. Aksakal, N. Colak, A. Demirbas, F. A. Ayaz, and N. Demirbas Vol 000
was recrystallized from an appropriate solvent to give the
desired compound.
169.91 (triazole C-3), 176.62 (triazole C-5), 181.10
(C=O). LC MS m/z (%): 607.69 ([M-HOO] 10), 321.37
+
4
-Benzyl-1-(piperidin-1-ylmethyl)-3-(thiomorpholinomethyl)-
H-1,2,4-triazole-5(4H)-thione (11a). Yield: 36% (method
), 54% (method 2), 53% (method 3); mp 51–52°C.
(20), 320.43 (100), 224.34 (45). Elemental analysis for
C H FN O S . Calculated (%), C: 58.38; H: 5.69; N:
15.37. Found (%), C: 58.41; H: 5.69; N: 15.33.
1
1
31
36
7 3 2
À1
1
FTIR (υmax, cm ): 3072, 3028 (ar-CH), 1286 (C=S). H
NMR (DMSO-d , δ ppm): 1.23 (bs, 2H, CH ), 1.46 (t,
H, 2CH , J = 4.0 Hz), 2.31 (t, 4H, 2CH , J= 4.0 Hz),
.65 (t, 4H, 2CH , J= 4.0Hz), 3.88 (s, 2CH + H O), 3.45
s, 2H, CH ), 5.04 (s, 2H, CH ), 5.34 (s, 2H, CH ), 7.19–
1-Ethyl-6-fluoro-4-oxo-7-(4-{[5-oxo-4-phenyl-3-(thiomor-
pholin-4-ylmethyl)-4,5-dihydro-1H-1,2,4-triazol-1-yl]methyl}
piperazin-1-yl)-1,4-dihydroquinoline-3-carboxylicacid (12b).
Yield: 49% (method 1), 82% (method 2), 93% (method 3);
6
2
4
2
2
2
2
2
2
À1
(
mp 250–251°C. FTIR (υmax, cm ): 3045 (ar-CH), 1707
2
2
2
1
3
1
7
.37 (m, 5H, arH). C NMR (DMSO-d , δ ppm): 23.96
(3C=O), 1495 (C=N). H NMR (DMSO-d , δ ppm): 1.44
6
6
(
(
[
CH ), 25.94 (2CH ), 27.10 (2CH ), 47.70 (CH ), 51.74
(s, 3H, CH ), 2.72 (bs, 4H, 2CH ), 2.85 (bs, 4H, 2CH ),
3.10 (m, 4H, 2CH ), 3.20 (s, 4H, 2CH ), 3.36 (s, 2H,
2 2
CH ), 4.57 (s, 2H, CH ), 4.69 (s, 2H, CH ), 6.96 (bs, 2H,
2 2 2
arH), 7.17 (bs, 1H, arH), 7.85–7.95 (bs, 2H, arH), 8.59
2
2
2
2
3
2
2
2CH ), 52.99 (CH ), 54.46 (2CH ), 70.14 (CH ), arC:
2
2
2
2
127.10 (2CH), 127.25 (CH), 127.78 (2CH), 136.55 (C)],
48.28 (triazole C-5), 169.67 (triazole C-3). LC MS m/z
1
+
(%): 404.33 ([M + 1] 60), 360.47 (45), 215.13 (100).
(s, 1H, quinolone CH), 15.18 (bs, 1H, OH, D O exch.).
2
13
Elemental analysis for C H N S . Calculated (%), C:
C NMR (DMSO-d , δ ppm): 14.53 (CH ), 27.68
2
0
29
5
2
6
3
5
9.52; H: 7.24; N: 17.35. Found (%), C: 59.55; H: 7.29;
(2CH ), 49.66 (2CH ), 52.42 (2CH ), 53.26 (2CH ),
2 2 2 2
N: 17.32.
61.64 (3CH ), arC: [106.31 (CH), 107.47 (C), 110.47–
2
3
-(Piperidin-1-ylmethyl)-5-(thiomorpholinomethyl)-1,3,4-
111.97 (d, CH, J= 15.0Hz), 112.54 (C), 114.42 (2CH),
118.56 (CH), 124.20 (2CH), 126.20 (C), 129.31 (C),
132.49 (C), 136.42 (CH), 148.29 (quinolone CH),
149.52–156.65 (d, C, J =713.0 Hz)], 165.31 (C=O),
168.25 (triazole C-3), 175.75 (triazole C-5), 181.91
oxadiazole-2(3H)-thione (11b).
8% (method 2), 93% (method 3); mp 223–224°C. FTIR
υmax, cm ): 3013 (ar-CH), 1295 (C=S). H NMR
DMSO-d , δ ppm): 1.32-1.63 (m, 10H, 5CH ), 2.59 (s,
H, 2CH ), 2.71 (s, 4H, 2CH ), 3.70 (s, 2H, CH ), 5.19
s, 2H, CH2). C NMR (DMSO-d , δ ppm): 23.59
CH ), 27.49 (2CH ), 28.41 (2CH ), 29.98 (2CH ), 52.47
CH ), 54.17 (2CH ), 67.69 (CH ), 161.20 (oxadiazole
Yield: 75% (method 1),
7
(
(
4
(
(
(
À1
1
6
2
+
(C=O). LC MS m/z (%): 607.69 ([M] 10), 310.53 (100),
2
2
2
1
3
224.34 (45). Elemental analysis for C H FN O S.
6
30 34
7 4
Calculated (%), C: 59.29; H: 5.64; N: 16.13. Found (%),
C: 59.31; H: 5.64; N: 16.23.
2
2
2
2
2
2
2
C-5), 179.46 (oxadiazole C-3). LC MS m/z (%): 316.21
7-(4-{[5-Oxo-4-phenyl-3-(thiomorpholin-4-ylmethyl)-4,5-
dihydro-1H-1,2,4-triazol-1-yl] methyl}-ciprofloxacin (12c).
+
([M + 2] 55), 218.09 (100), 159.07 (10), 116.15 (25).
Elemental analysis for C H N OS . Calculated (%): C:
Yield: 78% (method 1), 53% (method 2), 97% (method
1
3
22
4
2
À1
4
9.65; H: 7.05, N: 17.82. Found (%): C: 49.62; H: 7.09;
3); mp 250–251°C. FTIR (υ , cm ): 3089 (ar-CH),
max
1
N: 17.81.
1719 (3C=O), 1464 (C=N). H NMR (DMSO-d , δ
6
7
-(4-{[4-Benzyl-5-oxo-3-(thiomorpholin-4-ylmethyl)-4,5-di-
ppm): 1.13 (s, 2H, CH ), 1.32 (s, 2H, CH ), 2.34 (bs, 4H,
2
2
hydro-1H-1,2,4-triazol-1-yl]methyl}piperazin-1-yl)-1-ethyl-6-
fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid (12a).
2CH ), 2.73 (s, 4H, 2CH ), 2.86 (s, 4H, 2CH ), 3.09 (s,
2 2 2
2H, CH ), 3.81 (s, 4H, 2CH ), 4.34 (bs,1H, CH), 4.69 (s,
2
2
Yield: 56% (method 1), 57% (method 2), 85% (method
2H, CH ), 7.46–7.59 (m, 3H, arH), 7.85–7.90 (m, 3H,
2
À1
3
); mp 244–245°C. FTIR (υmax, cm ): 3051, 2956 (ar-
arH), 7.95 (s, 1H, arH), 8.65 (s, 1H, quinolone CH),
13
CH), 1719 (C=O), 1629 (C=O), 1495 (C=N), 1257
15.70 (bs, 1H, OH, D O exch.). C NMR (DMSO-d , δ
2
6
1
(
C=S). H NMR (DMSO-d , δ ppm): 1.41 (t, 3H, CH ,
ppm): 8.15 (2CH ), 27.60 (2CH ), 36.09 (CH), 49.73
2 2
6
3
J =8.0 Hz), 2.28 (bs, 4H, 2CH ), 2.50 (bs, 4H, 2CH2
(2CH ), 51.08 (2CH ), 53.50 (2CH ), 66.19 (CH ), 80.09
2 2 2 2
2
+
DMSO-d ), 2.70–2.91 (m, 8H, 4CH ), 4.13 (s, 2H
(CH ), arC: [106.82 (CH), 107.20 (C), 111.12–111.61 (d,
6
2
2
CH ), 4.59 (d, 2H, CH , J = 8.0Hz), 5.18 (s, 2H, CH ),
CH, J = 49.0Hz), 118.98 (C), 127.69 (2CH), 128.78
(CH), 129.41 (2CH), 133.92 (C), 139.71 (C), 143.29 (C),
145.78–148.23 (d, C, JC-F = 245.0 Hz)], 152.45
(quinolone CH), 154.02 (C=O), 154.72 (triazole C-3),
166.36 (triazole C-5), 176.77 (C=O). LC MS m/z
2
2
2
5
.35 (s, 2H, CH ), 7.18–7.34 (m, 6H, arH), 7.91 (d, 1H,
2
1
3
arH, J =8.0 Hz), 8.95 (s, 1H, quinolone CH). C NMR
(
(
(
[
DMSO-d , δ ppm): 14.80 (CH ), 27.08 (2CH ), 47.82
6 3 2
2CH ), 48.42 (CH ), 49.51 (CH ), 50.05 (2CH ), 53.04
2
2
2
2
+
CH ), 54.47 (CH ), 69.02 (CH ), 79.95 (CH ), arC:
(%): 619.71 ([M] 10), 320.53 (100). Elemental
2
2
2
2
106.43 (CH), 107.54 (C), 111.51–111.73 (d, CH,
analysis for C H FN O S. Calculated (%), C: 60.08;
31
34
7 4
J =23.0 Hz), 119.71–119.89 (d, C, J= 18.0Hz), 127.12
2CH), 127.80 (CH), 128.87 (2CH), 137.49–137.62 (d,
H: 5.53; N: 15.82. Found (%), C: 60.31; H: 5.54; N:
15.23.
(
C, J =11.3 Hz), 141.10 (C), 148.58–148.96 (d, C,
J =38.0 Hz), 148.99 (CH), 154.93–160.11 (d, C,
J =518.0 Hz)], 148.99 (quinolone CH), 166.56 (C=O),
7-(4-{[5-(Thiomorpholin-4-ylmethyl)-2-thioxo-1,3,4-oxadia-
zol-3(2H)-yl]methyl}norfloxacine (12d).
Yield: 70%
(method 1), 62% (method 2), 88% (method 3); mp 210–
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2016
Antioxidant Capacity, Biological Activity, and Molecular Docking Studies of
New Hybrid Compounds
À1
+
2
1
1
2
3
2
11°C. FTIR (υmax, cm ): 3424 (OH), 3049 (ar-CH),
535.49, ([M +1] 10), 291.34 (39), 279.45 (45), 174.33
1
723 (2C=O), 1250 (C=S). H NMR (DMSO-d , δ ppm):
.41 (s, 2H, CH ), 2.61 (s, 4H, 2CH ), 2.73 (bs, 4H,
(100). Elemental analysis for C H N O S . Calculated
(%), C: 51.66; H: 5.65; N: 15.72. Found (%), C: 51.63;
H: 5.67; N: 15.90.
3,3-Dimethyl-7-oxo-6-({[5-oxo-4-phenyl-3-(thiomorpholin-4-
ylmethyl)-4,5-dihydro-1H-1,2,4-triazol-1-yl]methyl}amino)-
penicillanic acid (13b).
6
23 30 6 3 3
3
2
CH ), 2.92 (s, 4H, 2CH ), 3.38 (s, 4H, 2CH +H O),
2
2
2
2
.74 (s, 2H, CH ), 4.58 (d, 2H, CH , J = 8.1Hz), 5.04 (s,
H, CH ), 7.16 (d, 1H, arH, J =8.1 Hz), 7.88 (d, 1H, arH,
J =12.0 Hz), 8.94 (s, 1H, quinolone CH). C NMR
2
2
2
13
Yield: 25% (method 1), 59%
(
DMSO-d , δ ppm): 14.80 (CH ), 27.67 (2CH ), 48.19
(method 2), 69% (method 3); mp 125–126°C. FTIR
6
3
2
À1
(CH ), 49.77 (2CH ), 50.91 (CH ), 52.68 (2CH ), 54.18
(KBr, υmax, cm ): 3318 (NH), 3063 (ar-CH), 1772
2
2
2
2
1
(
2CH ), 69.89 (CH ), arC: [106.61 (CH), 107.83 (2C),
(3C=O), 1536 (C=N). H NMR (DMSO-d , δ ppm): 1.38
2
2
6
1
11.91 (CH), 119.71 (C), 137.57 (C), 145.93–151.88 (d,
(s, 3H, CH ), 1.51 (s, 3H,CH ), 2.30 (bs, 4H, 2CH ),
3
3
2
C, JC-F = 594.0Hz), 148.99 (quinolone CH), 154.58
2.43 (bs, 4H, 2CH ), 2.86 (s, 2H, CH ), 3.51 (bs, 2H,
2 2
(
C=O), 166.45 (oxadiazole C-5), 176.61 (C=O), 178.57
CH +H O), 5.22 (s, 2H, 2CH), 5.30 (s, 1H, CH), 7.18
2 2
(d, 2H, arH, J = 7.1 Hz), 7.22 (t, 1H, arH, J= 6.8 Hz), 7.30
+
(oxadiazole C-3). LC MS m/z (%): 560.69 ([M] 25),
2
21.37 (100). Elemental analysis for C H FN O S .
(d, 2H, arH, J =7.3 Hz), 7.96 (s, 1H, NH, D O exch.),
2
4
29
6
4
2
2
13
Calculated (%), C: 52.54; H: 5.33; N: 15.32. Found (%),
C: 52.54; H: 5.29; N: 15.33.
13.51 (bs, 1H, OH). C NMR (DMSO-d , δ ppm): 27.12
6
(2CH ), 36.21 (2CH ), 46.66 (2CH ), 54.44 (2CH ),
2
3
2
2
7
-(4-{[5-(Thiomorpholin-4-ylmethyl)-2-thioxo-1,3,4-
69.16 (CH), 70.96 (C), 71.19 (2CH), arC: [127.21 (2CH),
127.71 (CH), 128.43 (2CH), 136.71 (C), 136,98 (triazole
C-5)], 149.55 (2C=O), 168.71 (triazole C-3). LC MS m/z
oxadiazol-3(2H)-yl]methyl}-ciprofloxacin (12e). Yield: 75%
method 1), 65% (method 2), 98% (method 3); mp
08–209°C. FTIR (υmax, cm ): 3029 (ar-CH), 1730
2C=O), 1263 (C=S). H NMR (DMSO-d , δ ppm): 1.21
(
2
(
À1
+
+
(%): 522.49, ([M +H O] 15), 504.34 ([M] 35), 179.45
2
1
(100). Elemental analysis for C H N O S . Calculated
6
22 28 6 4 2
(
2
s, 2H, CH ), 1.61 (s, 2H, CH ), 2.61 (bs, 4H, 2CH ),
.73 (s, 4H, 2CH ), 2.92 (s, 4H, 2CH ), 3.38 (s, 4H,
2 2
(%), C: 52.36; H: 5.59; N: 16.65. Found (%), C: 52.63;
H: 5.57; N: 16.60.
2
2
2
2
CH +H O), 3.75 (s, 2H, CH ), 4.58 (s, 2H, CH ), 5.04
3,3-Dimethyl-7-oxo-6-({[2-thioxo-5-(thiomorpholin-4-ylme-
2
2
2
2
(
bs, 1H, CH), 7.16 (d, 1H, arH, J =8.1Hz), 7.89 (d, 1H,
thyl)-1,3,4-oxadiazol-3(2H)-yl]methyl}amino)-penicillanic acid
1
3
arH, J =8.1 Hz), 8.94 (s, 1H, quinolone CH). C NMR
(13c).
Yield: 73% (method 1), 40% (method 2), 80%
À1
(
(
(
DMSO-d , δ ppm): 11.80 (2CH ), 27.67 (2CH ), 48.21
(method 3); mp 110–111°C. FTIR (υmax, cm ): 3324
(NH), 3049 (ar-CH), 1723 (2C=O), 1251 (C=S).
6
2
2
1
2CH ), 52.18 (2CH ), 54.18 (2CH ), 58.89 (CH ), 69.83
CH ), arC: [106.61 (CH), 107.83 (C), 111.96 (CH),
H
2
2
2
2
NMR (DMSO-d , δ ppm): 1.33 (s, 3H, CH ), 1.41 (s,
2
6
3
1
19.71 (C), 137.57 (C), 145.93 (C), 148.94 (quinolone
CH), 151.88-154.58 (d, C, JC-F = 270.0Hz)], 166.46
C=O), 176.61 (oxadiazole C-3), 178.67 (oxadiazole C-
3H, CH ), 2.34 (bs, 4H, 2CH ), 2.43 (bs, 4H, 2CH ),
3 2 2
2.86 (s, 2H, CH ), 3.69 (bs, 2H, CH ), 5.22 (s, 2H,
2
2
13
(
2CH), 5.31 (s, 1H, CH). C NMR (DMSO-d , δ ppm):
6
5
2
), 182.46 (C=O). LC MS m/z (%): 560.69 ([M] + 25),
27.13 (2CH ), 36.29 (2CH ), 46.56 (2CH ), 54.41
2
3
2
21.37 (100). Elemental analysis for C H FN O S .
(2CH ), 69.16 (CH), 70.96 (C), 71.19 (2CH), 166.63
2
5
29
6
4
2
2
Calculated (%), C: 53.56; H: 5.21; N: 14.99. Found (%),
C: 53.41; H: 5.29; N: 14.93.
(2C=O), 170.45 (oxadiazole C-3), 176.63 (oxadiazole C-
5). LC MS m/z (%): 445.69 ([M] 45), 220.37 (100).
+
6
-({[4-Benzyl-3-(thiomorpholin-4-ylmethyl)-5-thioxo-4,5-di-
Elemental analysis for C H N O S . Calculated (%), C:
1
6 23 5 4 3
hydro-1H-1,2,4-triazole-1-yl]methyl}amino)-penicillanic acid
43.13; H: 5.20; N: 15.72. Found (%), C: 43.54; H: 5.29;
N: 15.73.
(
(
13a). Yield: 45% (method 1), 67% (method 2), 67%
method 3); mp 175–179°C. FTIR (KBr, υmax, cm ):
À1
7-({[4-Benzyl-3-(thiomorpholin-4-ylmethyl)-5-thioxo-4,5-
dihydro-1H-1,2,4-triazol-1-yl]methyl}amino)-cephalosporanic
acid (14a). Yield: 45% (method 1), 30% (method 2), 60%
3
300 (NH), 3063, 3031 (ar-CH), 1772 (C=O), 1639
1
(C=O), 1636 (C=N), 1287 (C=S). H NMR (DMSO-d , δ
6
À1
ppm): 1.38 (s, 3H, CH ), 1.51 (s, 3H,CH ), 2.30 (bs, 4H,
(method 3); mp 109–110°C. FTIR (υmax, cm ): 3301
3
3
2
CH ), 2.47 (bs, 4H, 2CH + DMSO-d ), 2.48 (s, 2H,
(NH), 2926 (ar-CH), 1770 (C=O), 1736 (C=O), 1644
2
2
6
1
CH ), 2.86 (s, 2H, CH ), 3.36 (bs, 2H, CH + H O), 5.27
(C=O), 1579 (C=N), 1225 (C=S). H NMR (DMSO-d , δ
2
2
2
2
6
(s, 2H, 2CH), 5.31 (s, 1H, CH), 7.19 (d, 2H, arH,
ppm: 2.00 (s, 3H, CH ), 2.31 (s, 4H, 2CH ), 2.48 (s, 4H,
3
2
J =7.2 Hz), 7.28 (t, 1H, arH, J= 6.8Hz), 7.31 (d, 2H, arH,
2CH ), 2.71 (s, 4H, 2CH ), 2.86 (s, 4H, 2CH ), 3.36–
2 2 2
J =7.2 Hz), 7.95 (s, 1H, NH, D O exch.), 13.81 (bs, 1H,
3.43 (m, 2H, OCH + H O), 5.28 (s, 1H, CH), 5.32 (s,
2 2
2
13
OH). C NMR (DMSO-d , δ ppm): 27.12 (2CH ), 36.24
1H, CH), 7.21–7.32 (m, 5H, arH), 7.93 (s, 1H, NH, D O
2
6
2
13
(
2CH ), 46.66 (2CH ), 53.19 (CH ), 54.44 (2CH ), 69.16
exch.). C NMR (DMSO-d , δ ppm): 21.48 (CH ),
3
2
2
2
6 3
(CH), 70.96 (C), 71.19 (2CH), arC: [127.24 (2CH),
27.12 (CH ), 31.25 (2CH ), 46.66 (CH ), 53.20 (CH ),
2 2 2 2
1
1
27.75 (CH), 128.83 (2CH), 136.61 (C + triazole C-5)],
49.55 (2C=O), 168.71 (triazole C-3). LC MS m/z (%):
(2CH ), 54.44 ( 2CH ), 62.52 (CH), 64.04 (CH ), 69.82
2 2 2
(OCH ), 71.45 (CH), arC: [127.23 (CH), 127.44 (CH),
2
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

S. Demirci, A. Mermer, G. Ak, F. Aksakal, N. Colak, A. Demirbas, F. A. Ayaz, and N. Demirbas Vol 000
1
+
27.75 (CH), 128.69 (CH), 128.83 (CH), 136.60 (C
triazole C-5), 148.62 (C)], 149.53 (2C=O), 162.74
(DMSO-d , δ ppm): 27.48 (2CH ), 51.23 (2CH ), 52.66
6 2 2
(2CH ), 54.44 (2CH ), 55.67 (CH ), 56.52 (OCH ),
2 2 2 3
(
(
1
C=O), 168.72 (triazole C-3), 175.68 (C=O). LC MS m/z
%): 591.39 ([M + 1] 21), 307.40 (33), 196.34 (45),
19.19 (100). Elemental analysis for C H N O S .
25 30 6 5 3
58.04 (CH ), arC: [126.32 (CH), 127.41 (CH), 128.05
(CH), 128.45 (2CH), 128.99 (CH), 129.61 (2CH), 131.01
(CH), 132.62 (C), 140.34 (C)], 162.74 (triazole C-5),
2
+
Calculated (%), C: 50.83; H: 5.12; N: 14.23. Found (%),
C: 50.85; H: 5.19; N: 14.22.
168.72 (C-OCH ), 169.68 (triazole C-3). LC MS m/z
(%): 481.37 ([M + 1] 25), 480.21 ([M] 55), 296.36 (100).
3
+
+
7
-({[5-Oxo-4-phenyl-3-(thiomorpholin-4-ylmethyl)-4,5-dihy-
Elemental analysis for C H N O S. Calculated (%), C:
2
5 32 6 2
dro-1H-1,2,4-triazol-1-yl] methyl}amino)-cephalosporanic acid
62.47; H: 6.71; N: 17.49. Found (%),C: 62.55; H: 6.79;
N: 17.42.
(
(
(
14b). Yield: 40% (method 1), 59% (method 2), 65%
method 3); mp 214–215°C. FTIR (υmax, cm ): 3302
À1
3-[(4-Phenylpiperazin-1-yl)methyl]-5-(thiomorpholin-4-
ylmethyl)-1,3,4-oxadiazole-2(3H)-thione (15b). Yield: 62%
(method 1), 78% (method 2), 85% (method 3); mp 171–
NH), 3026 (ar-CH), 1772, 1746 (4C=O), 1644 (C=O),
1
1
565 (C=N). H NMR (DMSO-d , δ ppm: 2.01 (s, 3H,
6
À1
CH ), 2.34 (s, 4H, 2CH ), 2.48 (s, 4H, 2CH ), 2.71 (s,
172°C. FTIR (υmax, cm ): 2994 (ar-CH), 1545 (C=N),
3
2
2
1
4
H, 2CH ), 2.86 (s, 2H, CH ), 3.53 (bs, 2H, OCH +
1236 (C=S). H NMR (DMSO-d , δ ppm: 2.60-2.62 (m,
2
2
2
6
H O), 5.28 (s, 1H, CH), 5.31 (s, 1H, CH), 7.25–7.32 (m,
4H, 2CH ), 2.73–2.75 (m, 4H, 2CH ), 2.82–2.85 (m, 4H,
2CH ), 3.11–3.14 (m, 4H, 2CH ), 3.74 (s, 2H, CH ),
2 2 2
2
2
2
1
3
5
H, arH), 7.83 (s, 1H, NH, D O exch.). C NMR
2
(
DMSO-d , δ ppm): 21.48 (CH ), 27.10 (CH ), 31.23
5.02 (s, 2H, CH ), 6.77 (t, 1H, arH, J = 7.2 Hz), 6.92 (d,
6
3
2
2
13
(2CH ), 46.66 (CH ), 54.44 (2CH ), 62.52 (CH), 64.04
2H, arH, J = 8.0 Hz), 7.17–7.21 (m, 2H, arH). C NMR
(DMSO-d , δ ppm): 27.38 (2CH ), 48.50 (2CH ), 49.99
2
2
2
(CH ), 69.82 (OCH ), 71.41 (CH), arC: [126.22 (CH),
2 2
6
2
2
1
1
1
27.44 (CH), 128.15 (CH), 128.65 (CH), 128.89 (CH),
36.61 (C+ triazole C-5), 148.62 (C)], 149.53 (2C=O),
62.74 (C=O), 168.72 (triazole C-3), 175.68 (C=O). LC
(2CH ), 52.45 (2CH ), 54.19 (CH ), 69.96 (CH ), arC:
2 2 2 2
[116.10 (2CH), 119.69 (CH), 129.28 (2CH), 151.30 (C)],
162.87 (oxadiazole C-5), 169.16 (oxadiazole C-3). LC
+
+
+
+
MS m/z (%): 561.37 ([M + 1] 25), 560.21 ([M] 50),
96.36 (100). Elemental analysis for C H N O S .
MS m/z (%): 561.37 ([M+ 1] 25), 560.21 ([M] 50),
1
196.36 (100). Elemental analysis for C H N OS .
2
4
28
6
6
2
18 25
5
2
Calculated (%), C: 51.41; H: 5.03; N: 14.99. Found (%),
C: 51.25; H: 5.09; N: 14.92.
Calculated (%), C: 55.21; H: 6.44; N: 17.89. Found (%),
C: 55.25; H: 6.49; N: 17.92.
7
-({[5-(Thiomorpholin-4-ylmethyl)-2-thioxo-1,3,4-oxadiazol-
4-Benzyl-2-{[(1-phenylpiperidin-4-yl)amino]methyl}-5-(thio-
morpholin-4-ylmethyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione
(16a). Yield: 40% (method 1), 55% (method 2), 60%
3
7
2
(2H)-yl]methyl}amino)-cephalosporanic acid (14c). Yield:
0% (method 1), 80% (method 2), 79% (method 3); mp
14–215°C. FTIR (υmax, cm ): 3402 (NH), 2934 (ar-
CH), 1768, 1732 (3C=O), 1545 (C=N) 1226 (C=S). H
À1
À1
(method 3); mp 89–90°C. FTIR (υmax, cm ): 3301
1
1
(NH), 2926 (ar-CH), 1579 (C=N), 1224 (C=S). H NMR
NMR (DMSO-d , δ ppm: 2.01 (s, 3H, CH ), 2.48 (s, 4H,
(DMSO-d , δ ppm: 1.98 (bs, 4H, 2CH ), 2.48 (bs, 4H,
6
3
6
2
2
2
5
CH ), 2.59 (s, 4H, 2CH ), 2.60 (s, 4H, 2CH ), 2.71 (s,
2CH ), 2.51 (bs, 4H, 2CH +DMSO-d ), 2.86 (bs, 4H,
2CH ), 3.30 (s, 2H, CH ), 3.35 (s, 1H, CH), 3.43 (s, 2H,
2 2
2
2
2
2 2 6
H, CH ), 3.68 (bs, 2H, OCH +H O), 5.22 (s, 1H, CH),
2
2
2
13
.31 (s, 1H, CH), 7.93 (s, 1H, NH, D O exch.).
C
CH ), 5.20 (s, 2H, CH ), 5.29 (s, 2H, CH ), 7.21–7.32
2 2 2
2
13
NMR (DMSO-d , δ ppm): 21.40 (CH ), 27.13 (CH ),
(m, 10H, arH). C NMR (DMSO-d , δ ppm): 27.48
6
3
2
6
3
1.27 (2CH ), 46.62 (CH ), 54.20 (2CH ), 61.52 (CH),
(2CH ), 29.12 (2CH ), 48.25 (2CH ), 50.66 (CH), 50.89
2
2
2
2
2
2
6
3.04 (CH ), 65.82 (OCH ), 70.41 (CH), arC: [132.43
(CH ), 51.45 (CH ), 54.65 (2CH ), 61.44 (CH ), 62.52
2 2 2 2
2
2
(
(
(
(
C), 135.45 (C)], 165.53 (C=O), 166.01 (C=O), 168.34
(CH ), arC: [125.23 (CH), 127.44 (2CH), 127.75 (2CH),
128.69 (2CH), 128.83 (CH), 129.60 (2CH), 138.62 (2C)],
2
oxadiazole C-5), 168.72 (oxadiazole C-3), 175.68
+
C=O). LC MS m/z (%): 561.37 ([M + 1] 25), 560.21
168.74 (triazole C-3), 168.72 (triazole C-5). LC MS m/z
+
+
[M] 50), 196.36 (100). Elemental analysis for
(%): 553.45 (100), 509.46 ([M+ 1] 69). Elemental
C H N O S . Calculated (%), C: 51.41; H: 5.03; N:
analysis for C H N S . Calculated (%), C: 63.74; H:
24
28
6
6
2
27 36 6 2
1
4.99. Found (%), C: 51.25; H: 5.09; N: 14.92.
7.13; N: 16.52. Found (%), C: 63.75; H: 7.19; N: 16.42.
4-Benzyl-2-[(4-methylpiperazin-1-yl)methyl]-5-(thiomorpho-
lin-4-ylmethyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione (16b).
Yield: 58% (method 1), 65% (method 2), 74% (method 3);
2
-{[4-(2-Methoxyphenyl)piperazin-1-yl]methyl}-4-phenyl-5-
(
(
thiomorpholin-4-ylmethyl)-2,4-dihydro-3H-1,2,4-triazol-3-one
15a). Yield: 70% (method 1), 75% (method 2), 80%
À1
1
À1
(
method 3); mp 114–115°C. FTIR (υmax, cm ): 3063
mp 150–151°C. FTIR (υmax, cm ): 3058 (ar-CH), 1288
1
(ar-CH), 1712, 1712 (C=O), 1535 (C=N). H NMR
(C=S). H NMR (DMSO-d , δ ppm): 2.11 (bs, 32H,
6
(
DMSO-d , δ ppm: 2.14 (s, 4H, 2CH ), 2.38 (s, 4H,
CH ), 2.31 (d, 4H, 2CH J = 2.8 Hz), 2.50 (bs, 2CH
6
2
3
2
2
2
2
7
CH ), 2.61 (s, 4H, 2CH ), 2.76 (s, 4H, 2CH ), 3.42 (s,
+ DMSO-d ), 2.68 (s, 2H, CH ), 3.83 (bs, 3CH + H O),
6 2 2 2
2
2
2
H, CH ), 3.80 (s, 3H, OCH ), 4.31 (s, 2H, CH ), 7.12–
3.45 (s, 2H, CH ), 5.07 (s, 2H, CH ), 5.35 (s, 2H, CH ),
2
3
2
2 2 2
13
1
3
.22 (m, 3H, arH), 7.43–7.48 (m, 6H, arH). C NMR
7.18–7.37 (m, 5H, arH). C NMR (DMSO-d , δ ppm):
6
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2016
Antioxidant Capacity, Biological Activity, and Molecular Docking Studies of
New Hybrid Compounds
2
5
7.34 (CH ), 46.48 (CH ), 47.99 (CH ), 50.48 (2CH ),
described by Kurihara et al. [32]. Lipase activity was
measured using 4-methylumbelliferyl oleate (4-MU oleate)
as a substrate. Briefly, compounds were mixed with
porcine pancreatic lipase 1:3 (v/v) and incubated for
30min. The microtiter plates containing 50 μL of 0.1-mM
4-MU oleate, 25 μL of diluted compound–lipase solution,
2
3
2
2
3.28 (CH ), 54.71 (2CH ), 55.25 (2CH ), 69.34 (2CH ),
2
2
2
2
arC: [127.27 (2CH), 128.03 (CH), 129.14 (2CH), 136.81
C)], 148.64 (triazole C-5), 169.99 (triazole C-3). LC MS
(
+
+
m/z: 389.69 (100), 417.60 ([M-1] 31), 419.60 ([M + 1]
1). Elemental analysis for C H N S . Calculated (%):
7
2
0 30 6 2
C: 57.38; H: 7.22; N: 20.08. Found (%): C: 57.39; H:
.31; N: 20.08.
25 μL of dH O, and assay buffer (13 mM of Tris–HCl,
2
7
150 mM of NaCl, and 1.3 mM of CaCl , pH 8.0) were
2
4
-Benzyl-2-(morpholin-4-ylmethyl)-5-(thiomorpholin-4-
incubated at 37°C for 20 min. After incubation, 0.1mL of
0.1-M (pH 4.2) citrate buffer was added to the reaction
mixture in order to stop the reaction. The amount of 4-
methylumbelliferone released by the lipase was measured
using a spectrofluorometer (SpectraMax M5, Molecular
Devices, Sunnyvale, CA) at an excitation wavelength of
355 nm and an emission wavelength of 460nm. The inhib-
itory activity of those compounds and orlistat (Xenical,
Hoffman, La Roche, Segrate, Italy), an inhibitor of pancre-
atic lipase, were measured at various concentrations. Re-
sidual activities were calculated by comparison against a
control without an inhibitor. The assays were performed
in triplicate. The IC50 value was determined as the concen-
tration of compound giving 50% inhibition of maximal
activity.
ylmehyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione (16c).
Yield: 55% (method 1), 90% (method 2), 66% (method
); mp 81–82°C. FTIR (υmax, cm ): 3072, 3028
ar-CH), 1286 (C=S). H NMR (DMSO-d , δ ppm): 1.99
s, 4H, 2CH ), 2.51 (s, 2H, CH + DMSO-d ), 3.46 (s,
H, 2CH ), 3.56 (s, 4H, 2CH ), 5.07 (s, 2H, CH ), 5.35
s, 2H, CH ), 7.19-7.37 (m, 5H, arH). C NMR (DMSO-
d , δ ppm): 27.09 (2CH ), 47.78 (CH ), 49.04 (2CH ),
3.02 (CH ), 54.46 (2CH ), 66.62 (2CH ), 69.23 (CH ),
arC: [127.05 (2CH), 127.79 (CH), 128.89 (2CH), 136.50
C), 148.51 (triazole C-5), 169.89 (triazole C-3). LC MS
m/z: 423.23 ([M + H O] , 10), 407.34 ([M + 2] 20),
06.40 ([M + 1] 66), 116.33 (100). Elemental analysis
À1
3
(
(
4
1
6
2
2
6
2 2 2
1
3
(
2
6
2
2
2
5
2 2 2 2
(
+
+
2
+
4
for C H N OS . Calculated (%): C: 56.27; H: 6.71; N:
19
27
5
2
1
7.27. Found (%): C: 56.25; H: 6.79; N: 17.28.
α-Glucosidase inhibition assay [33].
α-Glucosidase
Biological activity studies
inhibition assay was performed spectrophotometrically. α-
Glucosidase from S. cerevisiae (Sigma-Aldrich) was
dissolved in phosphate buffer (pH6.8, 50mM). Test
compounds were dissolved in DMSO. To 96-well
microtiter plates were added 20 μL of test sample, 20 μL
of enzyme (20 mU/mL), and 135 μL of buffer This was
then incubated for 15min at 37°C. After incubation,
25 μL of p-nitrophenyl-α-D-glucopyranoside (2mM,
Sigma Aldrich) was added, and change in absorbance
was monitored for 20min at 400 nm. The test compound
was replaced by DMSO (10% final) as the control.
Acarbose (Sigma-Aldrich) was used as a standard
inhibitor. The assays were performed in triplicate. The
IC50 value was determined as the concentration of
compound giving 50% inhibition of maximal activity.
Antimicrobial activity.
The test microorganisms were
obtained from the Hifzissihha Institute of Refik Saydam
Ankara, Turkey) and consisted of E. coli ATCC 35218,
Y. pseudotuberculosis ATCC 911, P. aeruginosa ATCC
3288, E. faecalis ATCC 29212, S. aureus ATCC 25923,
(
4
B. cereus 709 Roma, M. smegmatis ATCC 607, C.
albicans ATCC 60193 and S. cerevisiae RSKK 251. Test
organisms are standard strains except for B.cereus and S.
cerevisiae. All the newly synthesized compounds were
weighed and dissolved in hexane to prepare stock
solution of 20,000 μg/mL.
The antimicrobial effects of the substances were tested
quantitatively in the respective broth media using double
microdilution, and MIC values (μg/mL) were determined.
The antibacterial and antifungal assays were performed in
Mueller-Hinton broth (Difco, Detroit, MI) at pH7.3 and
in buffered yeast nitrogen base (Difco, Detroit, MI) at
pH 7.0, respectively. The microdilution test plates were
incubated for 18–24 h at 35°C. Brain heart infusion broth
Urease inhibition assay [34].
Reaction mixtures
comprising 25 μL of Jack Bean urease, 55 μL of buffer
(0.01 M of K HPO , 1 mM of EDTA, and 0.01 M of LiCl,
2
4
pH 8.2), and 10mM urea were incubated with 5 μL of the
test compounds at room temperature for 15min in
microtiter plates. The production of ammonia was
measured following the indophenol method and was used
to determine the urease inhibitory activity. The phenol
reagent (45 μL, 1% w/v phenol and 0.005% w/v sodium
nitroprusside) and alkali reagent (70 μL, 0.5% w/v
sodium hydroxide and 0.1% v/v NaOCl) were added to
each well. Increasing absorbance at 625nm was
measured after 20min, using a microplate reader
(SpectraMax M5, Molecular Devices). The percentage
(
Difco, Detriot, MI) was used for M. smegmatis and
incubated for 48–72 h at 35°C [31]. Ampicillin (10 μg)
and fluconazole (5 μg) were used as standard antibacterial
and antifungal drugs, respectively. Dimethylsulfoxide at a
dilution of 1:10 was used as the solvent control. The
results obtained are presented in Table 3.
Pancreatic lipase inhibition [32]. The inhibitory effects
of these compounds were evaluated against porcine
pancreatic lipase (AppliChem, Darmstadt, Germany)
(15 ng/mL). Lipase activity assay was performed as
inhibition
was
calculated
from
the
formula
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

S. Demirci, A. Mermer, G. Ak, F. Aksakal, N. Colak, A. Demirbas, F. A. Ayaz, and N. Demirbas Vol 000
1
00–(ODtestwell/ODcontrol) ×100. Thiourea was used as the
means of the force field AMBER99. Charges on the
enzyme and inhibitors were assigned using the force field
AMBER99 and force field MMF94X, respectively. The
active sites of enzymes were identified by the site finder
application in MOE. Triangle matcher algorithm and two
rescoring functions, London dG and GBVI/WSA dG, were
used to produce 20 poses of each ligand. All poses gener-
ated with docking were analyzed, and the best-scored pose
for each compound was selected for further investigation
of interactions with the corresponding enzyme.
standard inhibitor. In order to calculate IC50 values,
different concentrations of synthesized compounds and
standard were assayed under the same reaction conditions.
Antioxidant activity studies
2
,2-Diphenyl-1-picrylhydrazyl radical scavenging activity.
The scavenging activity of different chemicals was
determined using the free radical DPPH, as described by
Blois [35]. A 100-μL chemical solution was mixed with
1
mL of freshly prepared methanolic DPPH solution. The
reaction mixture was incubated for 30min at room
temperature in the dark and was then measured at
Statistical analysis. Statistical analyses were carried out
using SPSS (IBM Corp. Released 2013. IBM SPSS
Statistics for Windows, Version 22.0. IBM Corp.,
Armonk, NY) one-way analysis of variance (ANOVA)
software. All treatments were performed using at least
three replicates, and significant differences between all
treatments were compared using Duncan’s Multiple
Range Test (P <0.05). A statistical package (XLSTAT
version 2014.6) using ADDINSOFT (Damrémont, Paris,
France) was used to perform principal component
analysis (PCA).
5
20nm. The activity was expressed as μmol Trolox
equivalent.
The ferric reducing ability of plasma. The FRAP was
measured using the method described by Benzie and
Strain [36] with some modification. To 100 μL of each
sample was added 2900μL of freshly prepared FRAP
reagent containing 300mM of acetate buffer (pH 3.6),
1
0mM of TPTZ (2,4,6-tripyridyle-s-triazine), and 20mM
of FeCl .6H O in proportions of 10:1:1 (v/v). The
3
2
mixture was incubated for 30 min at 37°C and measured
at 593nm. The values were expressed as μmol of Trolox/g.
Cupric ion reducing antioxidant capacity. The CUPRAC
was measured following the procedure described by Apak
et al. [37] with some modification. Briefly, 100 μL of each
chemical solution was mixed with 900 μL of bidistilled
water, 1mL of acetate buffer solution (1 mM, pH: 7.0),
Acknowledgments. The support provided by Scientific and
Technological Research Council of Turkey (TUBITAK, Project
no: 113Z181) is greatly appreciated.
REFERENCES AND NOTES
1
mL of CuCl2 (10 mM), and 1 mL of 7.5-mM
neocuproine to a final volume of 4mL. The reaction
mixture was then incubated in the dark for 30min at
room temperature, and the absorbance of the reaction
mixture was measured at 450nm against a water blank.
Trolox was used as the standard calibration curves, and
the results were expressed as μmol Trolox equivalent/g.
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[
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Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2016
Antioxidant Capacity, Biological Activity, and Molecular Docking Studies of
New Hybrid Compounds
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Journal of Heterocyclic Chemistry
DOI 10.1002/jhet