New pyrazoline derivatives and their antidepressant activity

Article abstract of DOI:10.1016/j.ejmech.2010.06.011

Some triazolo-pyrazoline derivatives were synthesized to investigate their potential antidepressant activities. The chemical structures of the compounds were elucidated by IR, NMR and FAB⁺-MS spectral data and elemental analyses. Antidepressant-like activities of the test compounds (100 mg/kg) were screened using both modified forced swimming and tail suspension tests. Rota-Rod test was performed for the examination of probable neurological deficits due to the test compounds, which may interfere with the test results. The test compounds in the series exhibited different levels of antidepressant activities when compared to reference drug fluoxetine. None of the test compounds changed motor coordination of animals when assessed in the Rota-Rod test. Therefore, experimental results in this study were not interfered with motor abnormalities. The study supports the antidepressant-like activities of various pyrazoline and/or triazole derivatives and suggests a possible serotonin related mechanism of action for the tested compounds.

Full text of DOI:10.1016/j.ejmech.2010.06.011

European Journal of Medicinal Chemistry 45 (2010) 4383e4387
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Short communication
New pyrazoline derivatives and their antidepressant activity
,
a
a
a
Zafer Asım Kaplancıklı a , Ahmet Özdemir , Gülhan Turan-Zitouni , Mehlika Dilek Altıntop ,
*
Özgür Devrim Can b
a Anadolu University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, Yunus Emre campus Tepebasi, 26470 Eskis¸ehir, Turkey
b Anadolu University, Faculty of Pharmacy, Department of Pharmacology, Yunus Emre campus Tepebasi, 26470 Eskis¸ehir, Turkey
a r t i c l e i n f o
a b s t r a c t
Article history:
Some triazolo-pyrazoline derivatives were synthesized to investigate their potential antidepressant
activities. The chemical structures of the compounds were elucidated by IR, NMR and FABþ-MS spectral
data and elemental analyses. Antidepressant-like activities of the test compounds (100 mg/kg) were
screened using both modified forced swimming and tail suspension tests. Rota-Rod test was performed
for the examination of probable neurological deficits due to the test compounds, which may interfere
with the test results. The test compounds in the series exhibited different levels of antidepressant
activities when compared to reference drug fluoxetine. None of the test compounds changed motor
coordination of animals when assessed in the Rota-Rod test. Therefore, experimental results in this study
were not interfered with motor abnormalities.
Received 4 March 2010
Received in revised form
27 May 2010
Accepted 6 June 2010
Available online 12 June 2010
Keywords:
Triazole
Pyrazoline
Antidepressant activity
Modified forced swimming test
Tail suspension tests
Rota-Rod
Ó 2010 Elsevier Masson SAS. All rights reserved.
1. Introduction
antidepressant action with relation to central serotonergic system
[13,14].
Depression is a serious disorder with estimates of lifetime
prevalence as high as 21% of the general population in some
developed countries [1]. Treatment for this disease is possible with
antidepressant medications and psychotherapy for some parts of
the patients [2]. Tricyclics, monoamine oxidase (MAO) inhibitors,
selective serotonin reuptake inhibitors, serotoninenorepinephrine
reuptake inhibitors, norepinephrineedopamine reuptake inhibi-
tors, norepinephrine reuptake inhibitors, serotonin modulators and
norepinephrineeserotonin modulators are the major antidepres-
sant drug classes used for the treatment of depressive disorders
[3,4]. Although antidepressants have been used in the clinic for
several decades, most of them are inadequate in efficiency and have
many serious adverse side effects. Therefore, studies for discovering
and developing new antidepressant drugs with greater effective-
ness and lower adverse effects are still desirable [5,6].
Like triazoles, some pyrazolines were also reported as potential
antidepressant agents. Several researchers reported MAO inhibi-
tory activity for various pyrazoline derivatives [15e17]. On the
other hand, our research group recently suggested the possible
involvement of serotonergic system for the antidepressant action of
some 2-pyrazoline derivative compounds [18].
In this present study, we aimed to obtain new compounds
containing both pyrazoline and triazole rings in the same structure.
For this purpose, we synthesized a system combining two biolabile
components, 2-pyrazolines and 1,2,4-triazole, and investigated
their potential antidepressant effects.
2. Chemistry
In the present work, 4-amino-5-[2-(4-hydroxyphenyl)ethyl]-
2,4-dihydro-3H-1,2,4-triazol-3-thione (1) has been synthesized by
reacting thiocarbohydrazide with 3-(4-hydroxyphenyl)propionic
acid. The 1-(2-thienyl)-3-aryl-2-propen-1-ones were prepared by
reacting 2-acetylthiophene with aromatic aldehyde [19]. The 1-(2-
thienyl)-3-aryl-2-propen-1-ones were treated with hydrazine
hydrate to obtain 5-aryl-3-(2-thienyl)-2-pyrazolines [19]. The
reaction of 3-(2-thienyl)-5-aryl-2-pyrazolines with chloroacetyl
chloride gave 1-(chloroacetyl)-3-(2-thienyl)-5-aryl-2-pyrazolines
(2aef) [20]. Treatment of equimolar quantities of triazole (1) with
There are some papers reporting the antidepressant-like activ-
ities of several 1,2,4-triazole derivative compounds [7e9]. Some
triazole derivative drugs are currently in clinical use to treat
depression such as nefazodone and etoperidone [10e12]. Both
nefazodone and etoperidone have been reported to show their
* Corresponding author. Tel.: þ90 222 335 05 80/37 79; fax: þ90 222 335 07 50.
E-mail address: zakaplan@anadolu.edu.tr (Z.A. Kaplancıklı).
0223-5234/$ e see front matter Ó 2010 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejmech.2010.06.011

4384
Z.A. Kaplancıklı et al. / European Journal of Medicinal Chemistry 45 (2010) 4383e4387
Table 1
1-(chloroacetyl)-3-(2-thienyl)-5-aryl-2-pyrazolines (2aef) resul-
ted in the formation of the title compounds (3aef) (Scheme 1).
Characterization data of compounds 3aef are given in Table 1.
Some characteristics of the compounds.
Comp.
R1
R2
M.p., ꢁ
C
Yield %
Mol. Formula
25H24N6O2S2
C25H23FN6O2S2
C25H23ClN6O2S2
C26H26N6O2S2
C27H29N7O2S2
C26H24N6O4S2
M.W.
3a
3b
3c
3d
3e
3f
H
F
Cl
CH3
N(CH3)2
OꢀCH2ꢀO
e
e
e
e
e
244e245
177e179
230e231
169e171
235e237
161e162
70
69
75
69
80
73
C
504
522
539
518
547
548
3. Results
The structures of compounds 1 and 3aef were confirmed by
elemental analyses, MS-FABþ, IR 1H NMR and 13C NMR spectral
data. All compounds gave satisfactory elemental analysis. Mass
spectra (MS (FAB)) of compounds showed M þ 1 peaks, in agree-
ment with their molecular formula. IR spectra of compounds 3aef
assigned to C4(HA). The multiplet at 3.80e4.05 ppm is readily
assignable to the C4(HB). attributable to the geminal coupling with
C4(HA) and the vicinal coupling with C5(Hx). The C5(Hx) proton of
pyrazoline appeared as multiplet at 5.45e5.70 ppm due to vicinal
coupling with the two magnetically non-equivalent protons of the
methylene group at position 4 of the pyrazoline ring [22]. The CH2
protons of acetyl group are observed at 4.35e4.65 ppm as multi-
plet, it should be noted that this anomaly arises from conforma-
tional differences [23]. The all derivatives showed NH2 and OH
protons as singlets at 5.85e6.00 and 9.15e9.25 ppm regions,
respectively. But only NH2 protons of 3a were observed as multi-
plet. All the other aromatic and aliphatic protons were observed at
expected regions.
showed NH, C]O and C]N, C]C bands at 3385e3108 cmꢀ1
,
1678e1670 cmꢀ1 and 1575e1283 cmꢀ1 regions, respectively.
In the 250 MHz 1H NMR spectrum of triazole 1, NH peak was
observed as singlet at 13.45 ppm. Therefore, it was proved that the
triazole was found in thionic form.
In the 250 MHz 1H NMR spectrum of compounds 3aef, the
CH2eCH2 protons appeared at 2.70e3.05 ppm. Among the three
multiplets of the ABX spin system of the pyrazoline ring, that
appearing at 3.10e3.30 ppm with multiplet attributable to the
vicinal [C4(HA)eC5(Hx)] and geminal [C4(HA)eC4(HB)] couplings, is
In the 13C NMR spectra of the compounds, the signal due to the
carbonyl carbon appears at 164e165 ppm. 13C NMR chemical shift
values of the carbon atoms at 43e44 ppm (C4), 59e60.31 ppm (C5)
and 151e152 ppm (C3) corroborate the 2-pyrazoline character
deduced from the 1H NMR data. The signals due to the triazole
carbons are observed at 150e151 ppm (C3) and 156e157 ppm (C5),
respectively. The compounds have a signal at 34e35.37 ppm due to
SeCH2 carbon. The other aromatic and aliphatic carbons were
observed at expected regions.
As shown in Table 2, reference drug fluoxetine (10 mg/kg) and
the test compounds (100 mg/kg) significantly shortened the
immobility times of mice in tail suspension test. Similarly, in MFST
fluoxetine and all test compounds shortened the immobility and
prolonged the swimming times without any change in the climbing
times of mice (Table 3). Test compounds 3a and 3c were more
effective than fluoxetine and some other derivatives as shown in
both tables. Neither fluoxetine, nor test compounds significantly
altered motor coordination parameters of mice, when compared to
the control values (data not shown).
O
H
N
H
N
+
OH
H2N
NH2
S
HO
H
S
N
N
N
N
N
SH
N
NH2
NH2
HO
HO
1
R2
R1
O
+
NaOH
S
S
R1
O
O
R2
S
4. Discussion
R2
R1
N
H2N-NH2 . H2O
N
H
Experimental data obtained from the tail suspension tests
exhibited that all tested triazolo-pyrazoline derivative compounds
showed statistically significant antidepressant-like activities as we
hypothesized. Besides, MFST results confirmed these findings for all
O
S
Cl
Cl
R2
N
N
N(C2H5)3
Table 2
Cl
Effects of fluoxetine (10 mg/kg) and test compounds (100 mg/kg) on immobility
times of mice in tail suspension test.
O
R1
2a-f
Comp.
Dose
Immobility time (s)
Control
Fluoxetine
e
116.4 ꢂ 4.6
10 mg/kg
100 mg/kg
100 mg/kg
100 mg/kg
100 mg/kg
100 mg/kg
100 mg/kg
90.6 ꢂ 5.0b
69.0 ꢂ 2.7c, #, **, z
S
3a
3b
3c
3d
3e
3f
K2CO3
78.8 ꢂ 3.6c
R2
R1
N
1
+ 2a-f
72.1 ꢂ 3.2c, #,
85.9 ꢂ 3.1c
93.5 ꢂ 4.3b
96.1 ꢂ 5.4a
*
, y
N
N
N
S
N
O
NH2
HO
Significance against control values ap < 0.05, bp < 0.01, cp < 0.001; significance
against fluoxetine #p < 0.05; significance against 3e *p < 0.05, **p < 0.01; signifi-
cance against 3f yp < 0.01, z p < 0.001, n ¼ 7.
3a-f
Scheme 1. Synthetic protocol of the title compounds.

Z.A. Kaplancıklı et al. / European Journal of Medicinal Chemistry 45 (2010) 4383e4387
4385
heated in an oil-bath at 160e170 ꢁC for 2 h. The fused mass thus
obtained was dispersed with hot water to obtain the triazole. The
product was recrystallized from methanol.
Table 3
Effects of fluoxetine (10 mg/kg) and test compounds (100 mg/kg) on climbing,
swimming and immobility times of mice in MFST.
IR: 3319 cmꢀ1, 3269 cmꢀ1, 3207 cmꢀ1 NH, 1612e1435 cmꢀ1
C]N, C]C, 1220 cmꢀ1, 1178 cmꢀ1 CeN, C]S.
Comp.
Dose
Climbing
time(s)
Swimming
time(s)
Immobility
time(s)
1H NMR (250 MHz) (DMSO-d6)
d (ppm): 2.85e2.90 (4H, m,
Control
Fluoxetine 10 mg/kg
e
56.8 ꢂ 2.6
95.0 ꢂ 9.7
102.2 ꢂ 7.5
47.8 ꢂ 3.7 148.5 ꢂ 10.4b
67.6 ꢂ 4.1b
CH2-triazole, phenol-CH2), 5.60 (2H, s, NH2), 6.65e7.00 (4H, m,
aromatic protons), 9.20 (1H, s, OH), 13.45 (1H, s, NH).
3a
3b
3c
3d
3e
3f
100 mg/kg 43.3 ꢂ 3.5 191.8 ꢂ 9.1c, #,**,y
100 mg/kg 48.8 ꢂ 5.8 153.3 ꢂ 8.7c
100 mg/kg 45.0 ꢂ 5.9 193.7 ꢂ 8.9c, #,**,z
100 mg/kg 54.4 ꢂ 3.5 147.8 ꢂ 7.8b
100 mg/kg 53.5 ꢂ 4.2 140.0 ꢂ 7.7a
100 mg/kg 54.1 ꢂ 3.4 144.7 ꢂ 8.2b
33.3 ꢂ 3.9c, ##,**,x
52.5 ꢂ 6.3c
40.5 ꢂ 5.7c, #,
58.4 ꢂ 5.1c
66.2 ꢂ 5.5b
69.9 ꢂ 6.2b
*
,y
For C10H12N4OS calculated: 50.83% C, 5.12% H, 23.71% N; found:
50.87% C, 5.10% H, 23.70% N. MS (FAB) [M þ 1]þ: m/z 237.
6.1.1.2. 1-(2-Thienyl)-3-aryl-2-propen-1-ones (Chalcones). A mixture
of 2-acetylthiophene (0.06 mol), aromatic aldehyde (0.06 mol) and
10% aqueous sodium hydroxide (10 mL) in ethanol (50 mL) was
stirred at room temperature for about 3 h. The resulting solid was
washed, dried and crystallized from ethanol [20].
Significance against control values ap < 0.05, bp < 0.01, cp < 0.001; significance
against fluoxetine #p < 0.05, ##p < 0.01; significance against 3e *p < 0.05, **p < 0.01;
significance against 3f yp < 0.05, zp < 0.01, xp < 0.001, n ¼ 7.
test compounds and provided additional information related to
possible mechanism of this activity. Decrease in immobility and
increase in swimming times of mice without any change in the
climbing durations indicated that the antidepressant-like effects
exhibited in this study may be probably related to serotonergic
mechanism rather than noradrenergic mechanism on CNS [24,25].
However, this consideration must be clarified with further detailed
studies.
Additionally, none of the test compounds was found to cause
significant changes on motor coordination of mice in Rota-Rod tests
when applied at 100 mg/kg dose. Therefore, represented experi-
mental results in this study were not affected by probable motor
abnormalities.
6.1.1.3. 3-(2-Thienyl)-5-aryl-2-pyrazolines. A solution of the appro-
priate chalcone (0.03 mol) and hydrazine hydrate (80%) (0.06 mol)
in ethanol (50 mL) was refluxed for 3 h. The reaction mixture was
cooled and kept at 0 ꢁC overnight. The resulting solid was recrys-
tallized from ethanol [20].
6.1.1.4. 1-(Chloroacetyl)-3-(2-thienyl)-5-aryl-2-pyrazolines (2aef).
The pyrazoline (0.02 mol) and triethylamine (0.02 mol) were dis-
solved in dry toluene (30 mL) with constant stirring. Later, the
mixture was cooled in an ice bath, and chloroacetyl chloride
(0.02 mol) was added drop wise with stirring. The reaction mixture
thus obtained was further agitated for 1 h at room temperature. The
precipitate was filtered; the solvent was evaporated to dryness [21].
In the series, unsubstituted derivative 3a and chlorine
substituted derivative 3c were more effective than the test
compounds 3e and 3f as well as 10 mg/kg dose of reference drug
fluoxetine, with respect to antidepressant action.
6.1.1.5. 1-[[4-amino-3-[2-(4-hydroxyphenyl)ethyl]-4H-1,2,4-triazol-
5-yl]thioacetyl]-3-(2-thienyl)-5-aryl-2-pyrazoline (3aef). A mixture
of the triazole (1) (10 mmol), pyrazolines (2aef) and anhydrous
potassium carbonate in acetone (20 mL) was mixed at room
temperature for 6 h. The mixture was filtered, the filtrate was
evaporated until dryness. The residue was washed with water and
recrystallized from ethanol.
5. Conclusion
In conclusion, this study supports the previous findings
reporting the antidepressant-like activities of various pyrazoline
and/or triazole derivatives [7e9,15e18] and suggests a possible
serotonin related mechanism of action for the tested compounds.
6.1.1.5.1. 1-[[4-Amino-3-[2-(4-hydroxyphenyl)ethyl]-4H-1,2,4-tri-
azol-5-yl]thioacetyl]-3-(2-thienyl)-5-phenyl-2-pyrazoline
3385e3110 cmꢀ1 NH,1670 cmꢀ1 C]O,1572e1288 cmꢀ1 C]N, C]C.
1H NMR (250 MHz) (DMSO-d6)
(ppm): 2.75e3.00 (4H, m,
3a. IR:
d
6. Experimental
CH2-CH2), 3.10e3.20 (1H, m, pyrazoline C4eHA), 3.90e4.05 (1H,
m, pyrazoline C4eHB), 4.45e4.65 (2H, m, COCH2), 5.55e5.70 (1H, m,
pyrazoline C5eHx), 5.90e6.00 (2H, m, NH2), 6.75e7.70 (12H,
m, aromatic protons), 9.15 (1H, s, OH).
6.1. Chemistry
All reagents were used as purchased from commercial suppliers
without further purification. Melting points were determined by
using an Electrothermal 9100 digital melting point apparatus and
were uncorrected (Electrothermal, Essex, UK). The compounds
were checked for purity by TLC on silica gel 60 F254. Spectroscopic
data were recorded on the following instruments: IR, Shimadzu 435
IR spectrophotometer (Shimadzu, Tokyo, Japan); 1H NMR, Bruker
250 MHz NMR spectrometer (Bruker Bioscience, Billerica, MA, USA)
and 13C NMR, Bruker Avance II 500 MHz NMR spectrometer (Bruker
Bioscience, Billerica, MA, USA) in DMSO-d6 using TMS as internal
standard; MS-FAB, VG Quattro mass spectrometer (Fisons Instru-
ments Vertriebs GmbH, Mainz, Germany), Elemental analyses were
performed on a Perkin Elmer EAL 240 elemental analyser (Per-
kineElmer, Norwalk, CT, USA).
13C NMR (125 MHz) (DMSO-d6)
d (ppm): 26.39 (CH2, phenol-
CH2), 31.60 (CH2, CH2-triazole), 35.07 (CH2, SeCH2), 43.24 (CH2,
pyrazoline C4), 60.31 (CH, pyrazoline C5), 115.42 (2CH, aromatic),
125.81 (2CH, aromatic), 127.64 (C, CH aromatic), 128.46 (CH,
aromatic), 128.99 (2CH, aromatic), 129.52 (2CH, aromatic), 130.39
(CH, aromatic), 131.30 (CH, aromatic), 134.04 (C, aromatic), 141.88
(C, aromatic),150.93 (C, triazole C3),151.51 (C, pyrazoline C3),155.89
(C, aromatic), 156.08 (C, triazole C5), 164.79 (C, C]O).
For C25H24N6O2S2 calculated: 59.50% C, 4.79% H, 16.65% N;
found: 59.54% C, 4.82% H, 16.68% N. MS (FAB) [M þ 1]þ: m/z 505.
6.1.1.5.2. 1-[[4-Amino-3-[2-(4-hydroxyphenyl)ethyl]-4H-1,2,4-tri-
azol-5-yl]thioacetyl]-3-(2-thienyl)-5-(4-florophenyl)-2-pyrazoline
3b. IR: 3380e3115 cmꢀ1 NH, 1675 cmꢀ1 C]O, 1570e1285 cmꢀ1
C]N, C]C.
1H NMR (250 MHz) (DMSO-d6)
d (ppm): 2.80e3.00 (4H, m, CH2-
6.1.1. General procedure for the synthesis of compounds
CH2), 3.15e3.25 (1H, m, pyrazoline C4eHA), 3.90e4.00 (1H, m,
pyrazoline C4eHB), 4.40e4.60 (2H, m, COCH2), 5.60e5.70 (1H, m,
pyrazoline C5eHx), 5.90 (2H, s, NH2), 6.70e7.80 (11H, m, aromatic
protons), 9.20 (1H, s, OH).
6.1.1.1. 4-amino-5-[2-(4-hydroxyphenyl)ethyl]-2,4-dihydro-3H-
1,2,4-triazole-3-thione (1). Equimolar mixture of thiocarbohy-
drazide (0.1 mol) and 3-(4-hydroxyphenyl)propionic acid was

4386
Z.A. Kaplancıklı et al. / European Journal of Medicinal Chemistry 45 (2010) 4383e4387
13C NMR (125 MHz) (DMSO-d6)
d
(ppm): 26.41 (CH2, phenol-
triazole C3), 151.65 (C, pyrazoline C3), 155.92 (C, aromatic), 156.07
(C, triazole C5), 164.55 (C, C]O).
CH2), 31.56 (CH2, CH2-triazole), 35.00 (CH2, SeCH2), 43.13 (CH2,
pyrazoline C4), 59.70 (CH, pyrazoline C5), 115.42 (2CH, aromatic),
115.63, 115.82 (2CH, aromatic), 128.07 (C, aromatic), 128.48 (3CH,
aromatic), 129.49 (2CH, aromatic), 130.42 (CH, aromatic), 131.34
(CH, aromatic), 134.04 (C, aromatic), 138.05 (C, aromatic), 150.89 (C,
triazole C3), 151.50 (C, pyrazoline C3), 155.91 (C, aromatic), 156.11
(C, triazole C5), 162.65 (C, aromatic), 164.88 (C, C]O).
For C27H29N7O2S2 calculated: 59.21% C, 5.34% H, 17.90% N;
found: 59.19% C, 5.34% H, 17.92% N. MS (FAB) [M þ 1]þ: m/z 548.
6.1.1.5.6. 1-[[4-Amino-3-[2-(4-hydroxyphenyl)ethyl]-4H-1,2,4-tri-
azol-5-yl]thioacetyl]-3-(2-thienyl)-5-(3,4-methylenedioxyphenyl)-2-
pyrazoline 3f. IR: 3376e3111 cmꢀ1 NH, 1677 cmꢀ1 C]O,
1560e1289 cmꢀ1 C]N, C]C.
For C25H23FN6O2S2 calculated: 57.46% C, 4.44% H, 16.08% N;
found: 57.45% C, 4.40% H, 16.10% N. MS (FAB) [M þ 1]þ: m/z 523.
6.1.1.5.3. 1-[[4-Amino-3-[2-(4-hydroxyphenyl)ethyl]-4H-1,2,4-tri-
azol-5-yl]thioacetyl]-3-(2-thienyl)-5-(4-chlorophenyl)-2-pyrazoline
3c. IR: 3365e3122 cmꢀ1 NH, 1672 cmꢀ1 C]O, 1575e1299 cmꢀ1
C]N, C]C.
1H NMR (250 MHz) (DMSO-d6)
d (ppm): 2.75e3.00 (4H, m,
CH2eCH2), 3.15e3.30 (1H, m, pyrazoline C4eHA), 3.85e3.95 (1H,
m, pyrazoline C4eHB), 4.35e4.45 (2H, m, COCH2), 5.45e5.55 (1H, m,
pyrazoline C5eHx), 5.90 (2H, s, NH2), 6.00e6.15 (2H, m, OeCH2eO),
6.70e7.85 (10H, m, aromatic protons), 9.25 (1H, s, OH).
13C NMR (125 MHz) (DMSO-d6)
d (ppm): 26.44 (CH2, phenol-
1H NMR (250 MHz) (DMSO-d6)
d
(ppm): 2.80e3.05 (4H, m,
CH2), 31.63 (CH2, CH2-triazole), 35.19 (CH2, SeCH2), 43.18 (CH2, pyr-
azoline C4), 60.09 (CH, pyrazoline C5),101.39 (CH2, OeCH2eO),106.41
(CH, aromatic), 108.59 (CH, aromatic), 115.45 (2CH, aromatic), 119.13
(CH, aromatic), 128.50 (C, CH, aromatic), 129.50 (2CH, aromatic),
130.39 (CH, aromatic), 131.23 (CH, aromatic), 134.19 (C, aromatic),
135.86 (C, aromatic),146.79 (C, aromatic),147.79 (C, aromatic),150.92
(C, triazole C3),151.59 (2C, pyrazoline C3, aromatic),156.10 (C, triazole
C5), 164.81 (C, C]O).
CH2eCH2), 3.20e3.30 (1H, m, pyrazoline C4eHA), 3.80e3.95 (1H,
m, pyrazoline C4eHB), 4.45e4.65 (2H, m, COCH2), 5.65e5.70 (1H, m,
pyrazoline C5eHx), 5.85 (2H, s, NH2), 6.75e7.90 (11H, m, aromatic
protons), 9.25 (1H, s, OH).
13C NMR (125 MHz) (DMSO-d6)
d (ppm): 26.42 (CH2, phenol-
CH2), 31.56 (CH2, CH2-triazole), 34.91 (CH2, SeCH2), 43.03 (CH2,
pyrazoline C4), 59.71 (CH, pyrazoline C5), 115.40 (2CH, phenol C3,
C5), 127.91 (C, aromatic), 128.46 (3CH, aromatic), 128.93 (2CH,
aromatic), 129.48 (2CH, aromatic), 130.43 (CH, aromatic), 131.34
(CH, aromatic), 132.19 (C, aromatic), 133.95 (C, aromatic), 140.80 (C,
aromatic), 150.88 (C, triazole C3), 151.48 (C, pyrazoline C3), 155.91
(C, aromatic), 156.10 (C, triazole C5), 164.90 (C, C]O).
For C26H24N6O4S2 calculated: 56.92% C, 4.41% H, 15.32% N;
found: 56.95% C, 4.44% H, 15.28% N. MS (FAB) [M þ 1]þ: m/z 549.
6.2. Pharmacology
For C25H23ClN6O2S2 calculated: 55.70% C, 4.30% H, 15.59% N;
found: 55.73% C, 4.34% H, 15.62% N. MS (FAB) [M þ 1]þ: m/z 539.
6.1.1.5.4. 1-[[4-Amino-3-[2-(4-hydroxyphenyl)ethyl]-4H-1,2,4-tri-
azol-5-yl]thioacetyl]-3-(2-thienyl)-5-(4-methylphenyl)-2-pyrazoline
3d. IR: 3372e3118 cmꢀ1 NH, 1678 cmꢀ1 C]O, 1562e1283 cmꢀ1
C]N, C]C.
6.2.1. Animals
Adult male Swiss albino mice (25e30 g) were used for the
experiments. The animals were housed in a room with controlled
temperature (24 ꢂ1 ꢁC) for 12 h light/12 h dark cycle. All animals
were acclimatized to the laboratory environment for at least 48 h
before the experiments. Food and water were allowed ad libitum.
The experimental protocols have been approved by the Local
Ethical Committee on Animal Experimentation of Eskis¸ ehir
Anadolu University, Turkey.
1H NMR (250 MHz) (DMSO-d6)
d (ppm): 2.70e3.00 (7H, m, CH2-
CH2 and CH3), 3.10e3.25 (1H, m, pyrazoline C4eHA), 3.85e4.00 (1H,
m, pyrazoline C4eHB), 4.45e4.60 (2H, m, COCH2), 5.65e5.70 (1H, m,
pyrazoline C5eHx), 5.95 (2H, s, NH2), 6.85e7.80 (11H, m, aromatic
protons), 9.15 (1H, s, OH).
6.2.2. Chemicals and administration of the compounds
Fluoxetine hydrochloride used in this study was purchased from
SigmaeAldrich Chemical Company, St. Louis, MO.
Fluoxetine (10 mg/kg), test compounds (100 mg/kg) and control
solution (sunflower oil) were applied via intraperitoneal (i.p) route
for three times; 24, 5 and 0.5 h before the applications of the tests
[18,26].
13C NMR (125 MHz) (DMSO-d6)
d (ppm): 21.02 (CH3), 26.44 (CH2,
phenol-CH2), 31.59 (CH2, CH2-triazole), 35.14 (CH2, SeCH2), 43.27
(CH2, pyrazoline C4), 60.10 (CH, pyrazoline C5), 115.40 (2CH,
aromatic),125.76 (2CH, aromatic),128.46 (C, aromatic),129.48 (2CH,
aromatic),129.51 (CH, aromatic),130.34 (CH, aromatic),131.23 (2CH,
aromatic), 131.33 (CH, aromatic), 134.12 (C, aromatic), 136.84
(C, aromatic), 138.96 (C, aromatic), 150.92 (C, triazole C3), 151.56
(C, pyrazoline C3),155.93 (C, aromatic),156.09 (C, triazole C5),164.73
(C, C]O).
6.2.3. Assessment of antidepressant activity
6.2.3.1. Tail suspension test. Antidepressant-like activity of the test
compounds was screened using the tail suspension test similar to
that described by Steru et al. [27]. Mice were dangled from their
tail using adhesive tape placed approximately 1 cm from the tip of
the tail attached to a applicator stick and hung approximately
30 cm above a table. Mice were considered immobile only when
they fail to make any struggling movements and hung passively.
Immobility time for each animal was scored by stopwatch during
the last 4 min of a 6 min test [28].
For C26H26N6O2S2 calculated: 60.21% C, 5.05% H, 16.20% N;
found: 60.24% C, 5.08% H, 16.24% N. MS (FAB) [M þ 1]þ: m/z 519.
6.1.1.5.5. 1-[[4-Amino-3-[2-(4-hydroxyphenyl)ethyl]-4H-1,2,4-tri-
azol-5-yl]thioacetyl]-3-(2-thienyl)-5-(4-dimethylaminophenyl)-2-
pyrazoline 3e. IR: 3370e3108 cmꢀ1 NH, 1674 cmꢀ1 C]O,
1571e1306 cmꢀ1 C]N, C]C.
1H NMR (250 MHz) (DMSO-d6)
d (ppm): 2.80e3.00 (10H, m,
CH2eCH2 and N(CH3)2), 3.10e3.25 (1H, m, pyrazoline C4eHA),
3.80e3.95 (1H, m, pyrazoline C4eHB), 4.35e4.50 (2H, m, COCH2),
5.45e5.50 (1H, m, pyrazoline C5eHx), 5.90 (2H, s, NH2), 6.65e7.80
(11H, m, aromatic protons), 9.20 (1H, s, OH).
6.2.3.2. Modified forced swimming test (MFST). Antidepressant-
like activity of the test compounds was screened using the
modified forced swimming test (MFST) as described before
[18,24,29,30]. The mice were forced to swim individually in
a glass cylinder (diameter, 12 cm; height, 30 cm) containing
20 cm of water at 25 ꢂ1 ꢁC. A 15-min pretest session was fol-
lowed 24 h later by a 5-min test session. Swimming, climbing
and immobility times of animals over 5-s intervals were recor-
ded by a stopwatch [30].
13C NMR (125 MHz) (DMSO-d6)
d (ppm): 26.45 (CH2, phenol-
CH2), 31.66 (CH2, CH2-triazole), 35.37 (CH2, SeCH2), 41.95 (2CH3, N
(CH3)2), 43.13 (CH2, pyrazoline C4), 60.02 (CH, pyrazoline C5), 112.79
(2CH, aromatic), 115.41 (2CH, aromatic), 126.69 (C, aromatic),
128.46 (CH, aromatic), 129.40 (2CH, aromatic), 129.47 (2CH,
aromatic), 130.24 (CH, aromatic), 131.12 (C, aromatic), 131.33 (CH,
aromatic), 134.30 (C, aromatic), 150.16 (C, aromatic), 150.93 (C,

Z.A. Kaplancıklı et al. / European Journal of Medicinal Chemistry 45 (2010) 4383e4387
4387
[10] W.E. Heydorn, Exp. Opin. Invest. Drugs 4 (1995) 131e137.
Following the training and the test sessions, the animals were
dried in a heated enclosure. The water was changed and the
cylinder rinsed with clean water after each animal to avoid the
influence of alarm substances.
[11] B. Cusack, A. Nelson, E. Richelson, Psychopharmacology 114 (1994) 559e565.
[12] A. O’Brien, Receptor Binding in Drug Research. CRC Press, FL-USA, 1986, pp.
151e166.
ꢀ
[13] E. Przegalinski, A. Lewandowska, J. Neural Transm. 46 (1979) 303e312.
[14] W.V. Vieweg, Drugs Today (Barc) 34 (1998) 845e853.
[15] E. Palaska, M. Aytemir, I.T. Uzbay, D. Erol, Eur. J. Med. Chem. 36 (2001)
539e543.
[16] N. Gökhan, A. Yes¸ ilada, G. Uçar, K. Erol, A.A. Bilgin, Arch. Pharm. 336 (2003)
362e371.
[17] F. Chimenti, A. Bolasco, F. Manna, D. Secci, P. Chimenti, A. Granese, O. Befani,
P. Turini, R. Cirilli, F. La Torre, S. Alcaro, F. Ortuso, T. Langer, Curr. Med. Chem.
13 (2006) 1411e1428.
[18] O.D. Can, U.D. Ozkay, Z.A. Kaplancıklı, Y. Oztürk, Arch. Pharm. Res. 32 (2009)
1293e1299.
6.2.3.3. Assessment of motor coordination in Rota-Rod test. With the
aim of investigating test compounds induced any changes in motor
coordination of the animals, Rota-Rod test was performed. Before
the experimental session, three trials were given for three consec-
utive days on the Rota-Rod apparatus (Ugo Basile 7560, Milano, Italy)
set at a rate of 16 revolutions/min. Mice that were able to remain on
the rod longer than 180 s were selected for the test [31,32].
[19] R.A. Kabli, A.A. Khalaf, M.T. Zimaity, A.M. Khalil, A.M. Kaddah, H.A. Al-Rifaie,
J. Indian Chem. Soc. 68 (1991) 47e51.
[20] A.A. Khalaf, R.A. Kabli, M.T. Zimaity, A.M. Khalil, A.M. Kaddah, H.A. Al-Rifaie,
Indian J. Chem. Sect. B 32B (1993) 1125e1129.
[21] G. Turan-Zitouni, A. Özdemir, K. Güven, Arch. Pharm. Chem. Life Sci. 338
(2005) 96e104.
[22] V. Vijayabaskar, S. Perumal, S. Selvaraj, A. Lycka, R. Murugan,
M. Balasubramanian, Magn. Reson. Chem. 37 (1999) 133e139.
[23] I. Ledneczki, P. Forgo, J.T. Kiss, I. ÁMolnárPálinkó, J. Mol. Struct. 834e836
(2007) 349e354.
6.2.4. Statistical analyses
The obtained experimental data were evaluated by the one-way
analysis of variance (ANOVA) followed by Tukey’s test as post hoc.
The results are presented as means ꢂ standard error of means
(SEM). Differences between data sets were considered as significant
when p value was less than 0.05.
[24] M.J. Detke, I. Lucki, Behav. Brain Res. 73 (1996) 43e46.
[25] J.F. Cryan, A. Markou, I. Lucki, Trends Pharmacol. Sci. 23 (2002) 238e245.
[26] S. Mora, R. Millán, H. Lungenstrass, G. Díaz-Véliz, J.A. Morán, M. Herrera-Ruiz,
J. Tortoriello, J. Ethnopharmacol. 106 (2006) 76e81.
[27] L. Steru, R. Chermat, B. Thierry, P. Simon, Psychopharmacology (Berl) 85
(1985) 367e370.
[28] D.M. Nielsen, G.J. Carey, L.H. Gold, Eur. J. Pharmacol. 499 (2004) 135e146.
[29] Y. Nakatomi, C. Yokoyama, S. Kinoshita, D. Masaki, H. Tsuchida, H. Onoe,
K. Yoshimoto, K. Fukui, Neurosci. Lett. 436 (2008) 167e170.
[30] M. Tanaka, G. Telegdy, Brain Res. Bull. 77 (2008) 301e305.
[31] Z. Tabarelli, M.A. Rubin, D.B. Berlese, P.D. Sauzem, T.P. Missio, M.V. Teixeira, A.
P. Sinhorin, M.A.P. Martins, N. Zanatta, H.G. Bonacorso, C.F. Mello, Braz. J. Med.
Biol. Res. 37 (2004) 1531e1540.
References
[1] M.L. Wong, J. Licinio, Nat. Rev. Neurosci. 2 (2001) 343e351.
[2] A. Little, Am. Fam. Physician 80 (2009) 167e172.
[3] C. Richard, M.D. Shelton, J. Clin. Psychiatry 5 (2003) 27e32.
[4] A.M. Koenig, M.E. Thase, Pol. Arch. Med. Wewn. 119 (2009) 478e486.
[5] O. Berton, E.J. Nestler, Nat. Rev. Neurosci. 7 (2006) 137e151.
[6] W. Zhu, S. Ma, R. Qu, D. Kang, Life Sci. 79 (2006) 749e756.
[7] J.M. Kane, M.W. Dudley, S.M. Sorensen, F.P. Miller, J. Med. Chem. 31 (1988)
1253e1258.
[8] M.C. Sarvà, G. Romeo, F. Guerrera, M. Siracusa, L. Salerno, F. Russo, A. Cagnotto,
M. Goegan, T. Mennini, Bioorg. Med. Chem. 10 (2002) 313e323.
[9] G. Turan-Zitouni, Z.A. Kaplancıklı, F.S. Kılıç, K. Erol, Boll. Chim. Farm. 141
(2002) 192e196.
[32] B. Adzu, S. Amos, I. Muazzam, U.S. Inyang, K.S. Gamaniel, J. Ethnopharmacol.
83 (2002) 139e143.