Synthesis, Spectral Characterization, Cytotoxic, and Antimicrobial Activities of Some Novel Heterocycles Utilizing 1,3-Diphenylpyrazole-4-carboxaldehyde Thiosemicarbazone

Article abstract of DOI:10.1002/jhet.3232

A new series of heterocycles was synthesized via the reaction of readily obtainable 1,3-diphenylpyrazole-4-carboxaldehyde thiosemicarbazone with many carbon electrophiles, for example, chloroacetic acid, chloroacetyl chloride, ethyl chloroacetate, dimethyl acetylenedicarboxylate, maleic anhydride, 3′-nitro-ω-bromoacetophenone, malonic acid, acetylacetone, ethyl benzoylacetate, arylidene malononitrile, and ethyl cyanoacetate in attempt to construct imidazolidinone, thiazolidinone, thiazole, and pyrimidine derivatives. The behavior of the titled compound towards hydrazine hydrate was investigated, in addition to the ring closure under different conditions. Also, the reactions with 2-chloroquinoline-3-carboxaldehyde and chromone-3-carboxaldehyde were discussed. The structures of all products obtained were substantiated from their analytical and spectral data. The antitumor and antimicrobial activities of the synthesized compounds were examined.

Full text of DOI:10.1002/jhet.3232

Month 2018
Synthesis, Spectral Characterization, Cytotoxic, and Antimicrobial Activ-
ities of Some Novel Heterocycles Utilizing 1,3-Diphenylpyrazole-4-
carboxaldehyde Thiosemicarbazone
Sayed K. Ramadan*
and Hanan A. Sallam
Chemistry Department, Faculty of Science, Ain Shams University, Abassia, Cairo 11566, Egypt
*
E-mail: sayed.karam2008@sci.asu.edu.eg
Received March 19, 2018
DOI 10.1002/jhet.3232
Published online 00 Month 2018 in Wiley Online Library (wileyonlinelibrary.com).
A new series of heterocycles was synthesized via the reaction of readily obtainable 1,3-diphenylpyrazole-4-
carboxaldehyde thiosemicarbazone with many carbon electrophiles, for example, chloroacetic acid,
0
chloroacetyl chloride, ethyl chloroacetate, dimethyl acetylenedicarboxylate, maleic anhydride, 3 -nitro-ω-
bromoacetophenone, malonic acid, acetylacetone, ethyl benzoylacetate, arylidene malononitrile, and ethyl
cyanoacetate in attempt to construct imidazolidinone, thiazolidinone, thiazole, and pyrimidine derivatives.
The behavior of the titled compound towards hydrazine hydrate was investigated, in addition to the ring closure
under different conditions. Also, the reactions with 2-chloroquinoline-3-carboxaldehyde and chromone-3-
carboxaldehyde were discussed. The structures of all products obtained were substantiated from their analytical
and spectral data. The antitumor and antimicrobial activities of the synthesized compounds were examined.
J. Heterocyclic Chem., 00, 00 (2018).
INTRODUCTION
RESULTS AND DISCUSSION
Pyrazole derivatives occupy a unique place in the
medicinal chemistry due to their extensive applications,
for example, antitumor [1–4], antioxidant [5], anti-
inflammatory [6], antimicrobial [7–10], and antiviral
activities [11–14]. Also, it has been documented that
newly synthesized pyrazole derivatives known as
potential anticancer agent due to their B-Raf inhibitory
activities with IC50 values in the nano levels, for
example, compounds I–IV (Fig. 1) [15,16]. 1,3-
Diphenylpyrazole-4-carboxaldehyde thiosemicarbazone
was utilized to construct several N-heterocycles [17].
Accordingly, it seemed interesting to carry out the
transformation of the titled compound into many N-
heterocycles bearing pyrazole moiety as a platform to
evaluate them as antitumor and antimicrobial agents.
Chemistry.
1,3-Diphenylpyrazole-4-carboxaldehyde
thiosemicarbazone 2 was easily synthesized from the
condensation of the titled aldehyde with
1
thiosemicarbazide [17]. The reaction of thiosemicarbazone
2 with chloroacetic acid was discussed under different
conditions. Therefore, when the reaction was carried out in
refluxing pyridine for 4 h, the thioxoimidazolidinone
derivative 3 was obtained in 74% yield. While the reaction
in refluxing ethanol containing anhydrous sodium acetate
led to the construction of thiazolidinone 4 in 79% yield
(Scheme 1). The structures of these products were inferred
from their analytical and spectral data. The infrared (IR)
spectrum of 3 revealed the characteristic absorption band
ꢀ
1
for the imidazolidinone C═O at ν1718 cm as well as
C═S at ν1248 cm . The H-NMR spectrum displayed a
ꢀ
1
1
©
2018 Wiley Periodicals, Inc.

S. K. Ramadan and H. A. Sallam
Vol 000
Figure 1. Structures of some potent pyrazole derivatives.
Scheme 1. Synthetic pathways for the formation of compounds 3–8.
ꢀ
1
1
singlet signal at δ 11.86 ppm integrated to one proton
C═O at ν1717 cm . The H-NMR provided a singlet at δ
exchangeable with D O, which is attributed to NH of
12.05 ppm integrated to one proton exchangeable with
2
imidazolidine moiety, in addition to a singlet at δ
D O attributable for NH of thiazolidine moiety, in
2
3
.89 ppm integrated to two protons due to CH protons.
addition to a singlet at δ 3.95 ppm due to CH protons.
2
2
The IR spectrum of 4 was devoid from νC═S and showed
the characteristic absorption band for the thiazolidinone
Interestingly, compounds 3 and 4 can also be prepared in
fairly good yields through alternative routes via the
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Synthesis, Spectral Characterization, Cytotoxic, and Antimicrobial Activities
reaction of thiosemicarbazone
2
with chloroacetyl
refluxing toluene for 5 h [18]. On the other hand,
0
chloride in the presence of triethylamine/dioxane and
ethyl chloroacetate in the presence of anhydrous
sodium acetate/ethanol, respectively, which can be proved
by melting point, mixed melting point, thin layer
chromatography (TLC), and IR.
refluxing
2
with 3 -nitro-ω-bromoacetophenone in
ethanolic solution containing anhydrous sodium acetate
brings about the formation of thiazole derivative 8. The
structure of 8 was substantiated from its analytical and
spectral data. The IR spectrum was lacked to νC═O and
ꢀ
1
1
The aldol condensation of the methylene group at
position-5 of thiazolidinone 4 with p-nitrobenzaldehyde
in refluxing AcONa/AcOH afforded α,β-unsaturated
carbonyl compounds 5a,b. The formation of 5b could be
interpreted upon acetylation via the reaction medium
used. The structures of compounds 5a,b were elucidated
from their analytical and spectral data. The IR spectrum
of compound 5a displayed a broad absorption band
showed νNH at 3396 cm . The H-NMR spectrum was
completely compatible with the assigned structure. All
the foregoing reactions are illustrated by Scheme 1.
The behavior of 4-oxothiazolidin-5-ylacetic acid 7
towards bis-nucleophiles was investigated. Thus, its
condensation with o-phenylenediamine and/or o-
aminothiophenol in refluxing methanol containing
catalytic amount of HCl afforded benzimidazole 10a
and/or benzothiazole 10b derivatives, respectively. The
structures of 10a,b were confirmed from the study of the
IR together with chemical evidence by direct comparison
with authentic samples prepared from the reaction of the
ꢀ
1
3
432 cm (νOH) indicating to its existence as a mixture
of lactam and lactim forms. The IR spectrum of 5b was
devoid from νNH and provided the characteristic
absorption bands of C═O groups at ν1715 and
ꢀ
1
1
673 cm due to mechanical coupling between the two
2,4-dioxothiazolidin-5-ylacetic
acid
11
with
1
carbonyl groups of imide linkage. The H-NMR spectrum
of 5b was lacked to any labile hydrogen and showed its
existence as a mixture of E-isomer and Z-isomer in a
ratio of 80:20%, respectively (cf. Experimental). The
presence of methyl group in compound 5b was approved
o-phenylenediamine and/or o-aminothiophenol in
refluxing methanol/HCl [19] (identity: melting point,
mixed melting point, TLC). The formation of the
products 10a,b could be proceed via the elimination of
two water molecules to form the nonisolable intermediate
9a,b followed by hydrolysis as depicted in Scheme 3.
The reaction of acid 7 with thionyl chloride led to the
formation of furothiazole derivative 13 via the formation
of the intermediate acid chloride 12 followed by the
1
by the appearance of singlet signal in its H-NMR
spectrum at δ 1.91 ppm integrated to three protons.
Dimethyl acetylenedicarboxylate condensed with
thiosemicarbazone 2 in refluxing methanol for 1 h to
furnish thiazolidinone 6 in 82% yield (Scheme 1). The IR
spectrum of compound 6 displayed the C═O of
elimination of HCl and H molecules (Scheme 3). The
2
structure of 13 was established from its analytical and
spectral data. The IR spectrum displayed νC═O at
ꢀ
1
α,β-unsaturated ester group at ν1713 cm in addition to
ꢀ
1
ꢀ1
C═O of thiazolidinone moiety at ν1700 cm . The
1734 cm attributable for α,β-unsaturated C═O group of
1
H-NMR spectrum showed singlet for NH proton at δ
furanone moiety, as well as the absence of any labile
1
12.78 ppm (exchangeable with D O) in addition to
hydrogens in its H-NMR spectrum (cf. Experimental).
2
methyl protons singlet at δ 3.81 ppm. The reaction could
be visualized to occur via the nucleophilic attack of the
The heterocyclic ring closure of thiosemicarbazone 2
under two different reaction conditions was studied.
Therefore, refluxing 2 in pyridine for 6 h led to the
construction of triazolethione 14. While refluxing in
terminal NH on the ester carbonyl group to eliminate
2
methanol molecule followed by ring closure (Scheme 2).
A facile synthesis of 4-oxothiazolidin-5-ylacetic acid 7
was provided by treatment of 2 with maleic anhydride in
an equimolar mixture of HCl/CH COOH for 2 h
3
afforded the fused heterocyclic compound, namely,
Scheme 2. A plausible mechanistic pathway for the formation of thiazolidinone 6.
Journal of Heterocyclic Chemistry
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S. K. Ramadan and H. A. Sallam
Vol 000
Scheme 3. Synthetic pathways for compounds 10a,b and 13.
1
1
(
,3-diphenyl-1,6-dihydropyrazolo[3,4-c]pyrazole
Scheme 4). The structures of these compounds were
substantiated from their analytical and spectral data. The
15
H-NMR spectrum of 14 indicated to its existence as a
mixture of thiolactam ⇌ thiolactim isomers in a ratio of
1:1. It displayed two NH protons at δ 9.30 and 8.55 ppm,
Scheme 4. Synthetic pathways for the formation of compounds 14, 15, and 17.
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Synthesis, Spectral Characterization, Cytotoxic, and Antimicrobial Activities
1
as well as SH proton at δ 8.85 ppm. The H-NMR spectrum
of 15 exhibited only one labile hydrogen at δ 13.20 ppm
attributable for NH of pyrazole moiety. The formation of
pyrazolopyrazole 15 could be explained via 1,5-endo-trig
cyclization by nucleophilic attack of the NH on C-5 of
pyrazole moiety followed by elimination of H─N═C═S
molecule.
mixed melting point, TLC). Presumably, the formation of
azine 17 could be achieved via Scheme 5.
This work extended to utilize thiosemicarbazone 2 to
construct pyrimidine heterocycles. Indeed, treatment of 2
with malonic acid in 5 ml of acetyl chloride afforded the
pyrimidinethione 18 (Scheme 6). The structure of 18 was
elucidated from its analytical and spectral data. The IR
spectrum provided the characteristic absorption bands of
Hydrazinolysis of
2
failed to produce the
ꢀ
1
hydrazinotriazole 16 and furnished a mixture of the
diheteryl azine 17 and thiosemicarbazide (cf. Scheme 4).
The structure of 17 was confirmed from the study of the
IR together with chemical evidence by direct comparison
with an authentic sample prepared from reaction of
C═O at ν1721 and 1672 cm attributable for ester and
amide groups, respectively, as well as NH group at
ꢀ
1
ꢀ1
1
ν3257 cm
and C═S group at ν1213 cm . The H-
NMR spectrum showed its existence as a mixture of E-
isomer and Z-isomer in a ratio of 1:1. Thus, it displayed
two singlets at δ 2.03 and 1.97 ppm each integrated to
1
,3-diphenylpyrazole-4-carboxaldehyde 1 with hydrazine
hydrate in refluxing ethanol [8] (identity melting point,
three protons attributable for CH protons, as well as two
3
singlets for NH proton at δ 10.84 and 10.79 ppm
exchangeable with D O (cf. Experimental).
2
Refluxing
2
with either acetylacetone, ethyl
Scheme 5. A suggested mechanistic pathway for the formation of
azine 17.
benzoylacetate or 2-(4-methoxybenzylidene) malononitrile
in refluxing sodium ethoxide solution led to the formation
of pyrimidine derivatives 19–21. The structures of
compounds 19–21 were demonstrated from their
analytical and spectral data. The IR spectra were devoid
from the characteristic absorption bands of νC═O group.
1
The H-NMR spectrum of 19 displayed two singlets at δ
2
.25 and 2.22 ppm each integrated to three protons
1
attributable for two methyl protons. The H-NMR
spectrum of 21 exhibited broad singlet at δ 6.50 ppm
integrated to two protons exchangeable with D O
2
attributable for NH protons. Interestingly, 6-hydroxy-5-
2
Scheme 6. Synthetic pathways for the formation of compounds 18–24.
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S. K. Ramadan and H. A. Sallam
Vol 000
ꢀ
1
cyanopyrimidine 22 was synthesized via a one-pot
cyclocondensation reaction between equivalent molar
quantities of thiosemicarbazone 2, ethyl cyanoacetate,
and p-anisaldehyde in refluxing sodium ethoxide solution
cm attributable for H-bonded NH, as well as C═O at
ꢀ
1
1
ν1662 cm . The H-NMR spectrum exhibited two
singlets at δ 11.30 and 10.50 ppm each integrated to one
proton attributable for two NH protons. The formation of
23 could be proceed via nucleophilic attack of the terminal
NH2 group in thiosemicarbazone 2 on position-2 of
quinoline nucleus to eliminate HCl molecule. Also, the
(Scheme 6). The structure of 22 was deduced from its
analytical and spectral data. The IR spectrum revealed the
absence of the characteristic absorption bands of νC═O
group and the appearance of νOH as well as νC≡N
reaction of chromone-3-carboxaldehyde with
2 in
ꢀ
1
ꢀ1
groups at 3445 cm and 2215 cm , respectively. The
refluxing ethanol underwent Michael addition on C-2 of
the chromonic moiety followed by dehydrogenation to
produce chromone derivative 24 (Scheme 7). The structure
of 24 was substantiated from its analytical and spectral
data. The IR spectrum of 24 showed a broad band at
1
H-NMR spectrum displayed a singlet for ─OH proton at
δ 11.31 ppm exchangeable with D O, as well as a singlet
2
at δ 3.79 ppm integrated to three protons attributable for
─
OCH protons (cf. Experimental).
On the other hand, reaction of 2-chloroquinoline-
3
ꢀ
1
ν3427 cm attributable for hydrogen bonded NH. The
H-NMR spectrum was completely consistent with the
1
3-carboxaldehyde with thiosemicarbazone 2 in refluxing
glacial acetic acid for 6 h afforded quinoline derivative 23.
The IR spectrum exhibited broad absorption band at ν3415
assigned structure (cf. Experimental).
Biological activity. Antimicrobial activity screening.
The reported biological activities of most of the hetero-
ring systems synthesized in the present investigation
promoted our interest to evaluate the antibacterial and
antifungal activities of synthesized compounds using the
diffusion disc method [20].
Scheme 7. Synthetic pathways for the formation of compounds 23
and 24.
The experiments were performed using test bacterial
organisms belonging to the Gram-positive and Gram-
negative, namely, Staphylococcus aureus and Escherichia
coli, respectively, as well as Aspergillus flavus and
Candida albicans as tested fungi; Ampicillin was used as
standard drug for bacteria, and amphotericin B was used as
standard drug for fungi. Preliminary screening of
the synthesized compounds and standard drugs was
performed at fixed concentration 20 mg/mL. Inhibition
was recorded by measuring the diameter of the inhibition
Table 1
Antibacterial and antifungal activity (as inhibition zone in mm diameter).
Antibacterial activity
Antifungal activity
ꢀ
+
Sample
Escherichia coli (G )
Staphylococcus aureus (G )
Aspergillus flavus
Candida albicans
3
4
6
7
8
12
13
12
23
8
14
12
11
20
5
11
5
5
15
7
0
0
0
18
0
1
1
1
1
1
2
2
2
2
2
3
4
5
8
9
0
1
2
3
4
7
5
0
0
12
10
12
0
11
12
14
16
18
25
—
13
10
12
2
12
13
13
15
19
21
—
11
13
11
3
10
14
15
13
15
—
16
10
11
10
0
8
10
12
10
11
—
21
Ampicillin
Amphotericin B
G: Gram reaction 0.0: no activity (inhibition zone less than 7 mm). 7–10: Weak activity 11–15: moderate activity. More than 15: strong activity.
Journal of Heterocyclic Chemistry
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Month 2018
Synthesis, Spectral Characterization, Cytotoxic, and Antimicrobial Activities
Figure 2. The effect of compounds on inhibition zone diameters against the tested organisms. [Color figure can be viewed at wileyonlinelibrary.com]
zoon at the end of 18 h for bacteria. Based on the results of
the inhibition zone, data in Table 1 and Figure 2 displayed
the inhibitory activities of the selected compounds. It has
been observed that compounds 7, 22, 23, and 24
exhibited a pronounced antimicrobial activity against all
the tested microorganisms compared with the reference
drugs, while compounds 3, 4, 6, 14, 18, 20, and 21
showed moderate activity. Compounds 13 and 19 showed
low activity. The carboxylic acid moiety in thiazolidinone
anticancer agent. The chemosensitivity responses of cell
lines to the new compounds are presented in Table 2. The
results obtained indicated that the tested compounds
exhibited good, moderate, or weak antiproliferative
activities against the tested cell lines. In general, the
compounds 7, 23, and 24 were found to be the most potent
derivatives against the two cell lines. The other
compounds were found to possess either moderate or
weak antiproliferative activities against the two cell lines.
7
increased both antibacterial and antifungal activities.
Anticancer activity
Structure activity relationships.
In vitro antiproliferative activity.
screening of the synthesized compounds was measured in
vitro using two different human cancer cell lines, namely,
hepatocellular carcinoma (HepG-2) and mammary gland
(MCF-7), using the standard MTT method [21].
Doxorubicin was selected as a standard reference
Table 2
Cytotoxic effect on human cell lines (HepG-2 and MCF-7).
a
In vitro cytotoxicity IC50 (μg/mL)
Sample
HepG-2
MCF-7
Doxorubicin
5.42 ± 0.31
78.03 ± 3.1
70.55 ± 0.2
81.15 ± 1.8
4.21 ± 2.9
64.23 ± 2.3
50.12 ± 0.9
67.62 ± 2.4
67.75 ± 0.2
45.84 ± 1.1
5.94 ± 0.5
4.17 ± 0.47
80.31 ± 2.2
75.21 ± 1.8
85.61 ± 1.2
5.33 ± 0.3
68.85 ± 4.1
49.80 ± 0.9
41.42 ± 0.3
58.62 ± 0.2
54.76 ± 0.6
4.81 ± 0.7
4.62 ± 0.2
3
4
6
7
8
1
1
1
2
2
2
4
5
8
0
3
4
5.61 ± 0.3
a
IC50 (lethal concentration of the sample that causes the death of 50% of
cells in 48 h): 1–10 (very strong), 11–20 (strong), 21–50 (moderate),
1–100 (weak), above 100 (noncytotoxic).
Figure 3. Structure activity relationships of the more potent compounds.
[Color figure can be viewed at wileyonlinelibrary.com]
5
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S. K. Ramadan and H. A. Sallam
Vol 000
CONCLUSION
(br., OH, enol form, NH), 3052 (aryl-H), 2956 (alkyl-H),
1
1
718 (C═O), 1640 (C═N), 1248 (C═S). H-NMR
Transformation of 1,3-diphenylpyrazole-4-carboxaldehyde
thiosemicarbazone into novel heterocycles, for example,
thiazolidinone, imidazolidine, thiazole, and pyrimidine
derivatives was carried out through a facile strategy and
evaluated for antitumor as well as antimicrobial activities.
The results showed that some of synthesized compounds
have promising activities.
(DMSO-d ): δH (ppm) 11.86 (s, 1H, NH, D O-
6
2
exchangeable), 8.94 (s, 1H, CH═N), 8.43 (s, 1H, C─H
pyrazolyl), 7.99–7.97 (d, 2H, Ph─C═N, J = 7.5 Hz),
7.87–7.85 (d, 2H, Ph─N, J = 8.1 Hz), 7.58–7.36 (m,
6H, Ar─H), 3.89 (s, 2H, CH ). MS (m/z, %): 361
2
+
.
(M , 42). Anal. Calcd for C H N OS (361.42): C,
1
9 15 5
63.14; H, 4.18; N, 19.38; S, 8.87. Found: C, 63.08; H,
4
.09; N, 19.40; S, 8.81.
Formation of thiazolidinone derivative 4. Method I:
EXPERIMENTAL
Reaction of 2 with chloroacetic acid.
A mixture of 2
(2 mmol), chloroacetic acid (2 mmol), and freshly
Chemistry.
All melting points were measured on a
prepared anhydrous sodium acetate (2 mmol) in an
absolute ethanol (20 mL) was heated under reflux for
3 h. The separated solid while hot was collected by
filtration and recrystallized from ethanol/dioxane (1:1)
to afford thiazolidinone 4 as white crystals, mp 306–
308°C, yield 79%.
Method II: Reaction of 2 with ethyl chloroacetate. To a
solution of 2 (2 mmol) and anhydrous sodium acetate
(2 mmol) in an absolute ethanol (20 mL), ethyl
chloroacetate (2 mmol) was added. The reaction mixture
was heated under reflux for 2 h. The precipitated solid
while hot was collected by filtration and recrystallized
from ethanol/dioxane (1:1) to afford 4, mp 306–308°C,
yield 83%.
Gallenkamp electric melting point apparatus and are
uncorrected. The Fourier transform infrared (FTIR)
spectra were recorded using potassium bromide disks on
Fourier Transform Infrared Thermo Electron Nicolet iS10
Spectrometer (Thermo Fisher Scientific Inc., Waltham,
MA, USA) at the Central Laboratory of Faculty of
1
Science, Ain Shams University. The H-NMR spectra
were run at 400 MHz on a GEMINI 400 BB NMR
Spectrometer (GEMINI, Manufacturing & Engineering
Inc., Anaheim, CA, USA) using tetramethyl silane
as internal standard in deuterated dimethylsulfoxide
(
DMSO-d ) at the Main Defense Chemical Laboratory,
6
Cairo. The mass spectra (MS) were recorded on a
Shimadzu GC–MS-QP-1000EX mass spectrometer
2-(((1,3-Diphenyl-1H-pyrazol-4-yl)methylene)hydrazono)
thiazolidin-4-one (4). IR (KBr) (ν, cm ): 3435 (br., OH,
ꢀ
1
(Shimadzu Scientific Instruments, Inc., USA) operating at
7
0 eV at the Micro-analytical Center of Cairo University.
lactim form), 3052 (aryl-H), 2961 (alkyl-H), 1717 (C═O),
1
The reactions were monitored by TLC using Merck
Kiesel gel 60F254 analytical sheets obtained from Fluka.
The biological activities were performed at Micro-
analytical Center of Mansoura University, Egypt.
1640 (C═N). H-NMR (DMSO-d
): δ
6
H
(ppm) 12.05 (s,
1H, NH, D O-exchangeable), 8.96 (s, 1H, CH═N), 8.50
2
(s, 1H, C─H pyrazolyl), 7.98–7.96 (d, 2H, Ph─C═N,
J = 7.5 Hz), 7.86–7.84 (d, 2H, Ph─N, J = 8.1 Hz), 7.57–
7
.36 (m, 6H, Ar─H), 3.95 (s, 2H, CH ). MS (m/z, %):
Formation of imidazolidine derivative 3. Method I:
2
+
.
Reaction of thiosemicarbazone 2 with chloroacetic acid.
A
361 (M , 33). Anal. Calcd for C19H N OS (361.42): C,
15 5
solution of pyrazolylthiosemicarbazone 2 (2 mmol) and
chloroacetic acid (2 mmol) in pyridine (10 mL) was
heated under reflux for 6 h and left overnight at room
temperature. The precipitated solid was collected by
filtration and recrystallized from ethanol/dioxane mixture
63.14; H, 4.18; N, 19.38; S, 8.87. Found: C, 63.03; H,
4.13; N, 19.35; S, 8.84.
Aldol
condensation
of
thiazolidinone
4
with
p-nitrobenzaldehyde. Equimolar mixture of thiazolidinone
derivative 4, p-nitrobenzaldehyde, and anhydrous sodium
acetate (2 mmol) in glacial acetic acid (10 mL) was
heated under reflux for 5 h. The separated solid while hot
was collected by filtration and recrystallized from dioxane
to give 5b, yield 51%. The residual mother liquor, after
the separation of 5b, was concentrated and then left to
cool at room temperature. The obtained precipitate was
filtered off and recrystallized from ethanol/dioxane
mixture (1:1) to afford 5a, yield 43%.
(1:1) to afford imidazolidine derivative 3 as white
crystals, mp 324–326°C, yield 74%.
Method II: Reaction of 2 with chloroacetyl chloride. To a
stirred solution of 2 (2 mmol) in dry dioxane (10 mL)
containing few drops of triethylamine, chloroacetyl
chloride (2 mmol) was added dropwise at room
temperature. Stirring was continued for 4 h. The reaction
mixture was then poured onto ice/cooled water with
stirring for 5 min. The precipitated solid was collected by
filtration and recrystallized from ethanol/dioxane mixture
2
-((-(1,3-Diphenyl-1H-pyrazol-4-yl)methylene)hydrazono)-5-
(-4-nitrobenzylidene)-thiazolidin-4-one (5a). Yellow crystals,
ꢀ
1
(1:1) to give 3, mp 324–326°C, yield 81%.
mp 258–260°C. IR (KBr) (ν, cm ): 3425 (br., OH,
3
-(((1,3-Diphenyl-1H-pyrazol-4-yl)methylene)amino)-2-
lactim form), 3051 (aryl-H), 1717 (C═O), 1640 (C═N).
ꢀ
1
1
thioxoimidazolidin-4-one (3). IR (KBr) (ν, cm ): 3421
H-NMR (DMSO-d ): δH (ppm) 11.89 (s, 1H, NH,
6
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0
D O-exchangeable), 8.94 (s, 1H, CH═N), 8.42 (s, 1H,
C─H pyrazolyl), 8.56–8.54 (d, 2H, p-nitrophenyl,
Reaction of thiosemicarbazone
2
with 3 -nitro-ω-
2
0
bromoacetophenone. A solution of 2 (2 mmol) and 3 -
nitro-ω-bromoacetophenone (2 mmol) in an absolute
ethanol (15 mL) containing anhydrous sodium acetate
2 mmol) was heated under reflux for 1 h. The precipitated
solid while hot was filtered off and recrystallized from
ethanol to afford thiazole derivative 8.
2-(2-((1,3-Diphenyl-1H-pyrazol-4-yl)methylene)hydrazinyl)-
-(3-nitrophenyl) thiazole (8). Yellow crystals, mp 198–
200°C, yield 81%. IR (KBr) (ν, cm ): 3396 (NH), 3058
J
=
8.1 Hz), 8.33–8.31 (d, 2H, p-nitrophenyl,
J = 8.0 Hz), 7.99 (s, 1H, CH═), 7.96–7.36 (m, 10H,
+.
(
Ar─H). MS (m/z, %): 494 (M , 24). Anal. Calcd for
C H N O S (494.52): C, 63.15; H, 3.67; N,
2
6 18 6 3
1
6
6.99; S, 6.48. Found: C, 63.04; H, 3.61; N, 17.01; S,
.41.
4
3
-Acetyl-2-(((1,3-diphenyl-1H-pyrazol-4-yl)methylene)
ꢀ
1
hydrazono)-5-(-4-nitrobenzylidene)-thiazolidin-4-one (5b).
ꢀ
1
1
Yellow crystals, mp 310–312°C. IR (KBr) (ν, cm ):
052 (aryl-H), 2938 (alkyl-H), 1716, 1673 (C═O), 1639
(aryl-H), 1628 (C═N). H-NMR (DMSO-d
6 H
): δ (ppm)
3
12.06 (s, 1H, NH, D O-exchangeable), 8.89 (s, 1H,
2
1
(C═N). H-NMR (DMSO-d ): (E-isomer and Z-isomer,
CH═N), 8.66 (s, 1H, m-nitrophenyl), 8.18 (s, 1H, C─H
pyrazolyl), 8.29–8.27 (d, 1H, m-nitrophenyl, J = 7.8 Hz),
8.15–8.13 (d, 1H, m-nitrophenyl, J = 8.4 Hz), 8.00–7.97
(d, 2H, Ph─C═N, J = 8.1 Hz), 7.81–7.79 (d, 2H,
Ph─N, J = 8.1 Hz), 7.72–7.67 (dd, 1H, m-nitrophenyl,
J = 8.1 & 7.8 Hz), 7.61 (s, 1H, C─H thiazolyl),
6
80:20%) δH (ppm) 10.18 and 9.99 (two singlets, 1H,
CH═N), 9.38 and 9.31 (two singlets, 1H, C─H
pyrazolyl), 8.93 and 8.47 (two singlets, 1H, CH═), 8.45–
8
2
.42 (d, 2H, p-nitrophenyl, J = 8.7 Hz), 8.29–8.27 (d,
H, p-nitrophenyl, J = 8.6 Hz), 8.01–7.38 (m, 10H,
+.
+.
Ar─H), 1.91 (s, 3H, CH ). MS (m/z, %): 536 (M , 27).
7.57–7.35 (m, 6H, Ar─H). MS (m/z, %): 466 (M , 71).
Anal. Calcd for C25H N O S (466.52): C, 64.37; H,
3
Anal. Calcd for C H N O S (536.56): C, 62.68; H,
18 6 2
2
8 20 6 4
3
1
.76; N, 15.66; S, 5.98. Found: C, 62.62; H, 3.73; N,
5.69; S, 5.91.
3.89; N, 18.01; S, 6.87. Found: C, 64.31; H, 3.83; N,
18.10; S, 6.82.
Reaction of thiosemicarbazone
acetylenedicarboxylate.
2
with dimethyl
Reaction of thiazolidinone 7 with o-phenylenediamine and/or
o-aminothiophenol. A solution of thiazolidinone-5-acetic
A solution of 2 (2 mmol) and
dimethyl acetylenedicarboxylate (2 mmol) in methanol
20 mL) was heated under reflux for 1 h. The precipitated
acid
7
(2 mmol) and o-phenylenediamine or
(
o-aminothiophenol (2 mmol) in methanol (15 ml)
containing few drops of conc. HCl was heated under reflux
for 1 h. The separated solid while hot was filtered off and
recrystallized from dioxane to afford benzimidazole 10a
and benzothiazole 10b derivatives, respectively. These
solid while hot was collected by filtration and
recrystallized from methanol/dioxane mixture (2:1) to
afford thiazolidinone derivative 6.
Methyl 2-(-2-((-(1,3-diphenyl-1H-pyrazol-4-yl)methylene)
hydrazono)-4-oxothiazolidin-5-ylidene) acetate (6).
1
compounds were identical in all respects (IR, HNMR, mp,
Yellow crystals, mp 300–302°C, yield 82%. IR (KBr)
mixed mp and TLC) with authentic samples prepared by
reacting of equimolar mixture of 2-(2,4-dioxothiazolidin-
5-yl) acetic acid 11 with o-phenylenediamine or
o-aminothiophenol in methanol/HCl.
ꢀ
1
(
2
ν, cm ): 3446 (br., OH, lactim form), 3056 (aryl-H),
986, 2954 (alkyl-H), 1713 (C═O ester), 1700 (C═O
1
thiazolidinone), 1636 (C═N). H-NMR (DMSO-d ): δ
6
H
(
ppm) 12.78 (s, 1H, NH, D O-exchangeable), 9.03 (s,
2
5-((1H-benzo[d]imidazol-2-yl)methyl)thiazolidine-2,4-
1
2
H, CH═N), 8.52 (s, 1H, C─H pyrazolyl), 8.01–7.98 (d,
H, Ph─C═N, J = 8.4 Hz), 7.89–7.86 (d, 2H, Ph─N,
dione (10a). White crystals, mp 244–246°C, yield 71%.
[Lit. [19] mp 244–246°C].
J = 7.5 Hz), 7.59–7.39 (m, 6H, Ar─H), 6.66 (s, 1H,
5
-(Benzo[d]thiazol-2-ylmethyl)thiazolidin-2,4-dione (10b).
+
.
CH═), 3.81 (s, 3H, CH ). MS (m/z, %): 431 (M , 54).
Yellow crystals, mp 172–174°C, yield 79%. [Lit. [19] mp
172–174°C].
3
Anal. Calcd for C H N O S (431.47): C, 61.24; H,
2
2 17 5 3
3
1
.97; N, 16.23; S, 7.43. Found: C, 61.21; H, 3.92; N,
6.29; S, 7.38.
Reaction of thiazolidinone (7) with thionyl chloride.
A
solution of thiazolidinone-5-acetic acid 7 (2 mmol) in
thionyl chloride (5 mL) was heated at 60°C for 5 h. The
reaction mixture was evaporated under vacuum. The
remained residue was recrystallized from dioxane to
produce furothiazole derivative 13.
Reaction of thiosemicarbazone 2 with maleic anhydride.
A suspension of 2 (2 mmol) and maleic anhydride
2 mmol) in dry toluene (15 mL) was heated under
(
reflux for 4 h. The reaction mixture was concentrated
and cooled to room temperature. The obtained solid
was collected by filtration and recrystallized from
ethanol/dioxane mixture (1:1) to afford thiazolidinone
derivative 7.
2
-((-(1,3-Diphenyl-1H-pyrazol-4-yl)methylene)hydrazono)
furo[2,3-d]thiazol-5(2H)-one (13).
>360°C, yield 43%. IR (KBr) (ν, cm ): 3058 (aryl-H),
Yellow crystals, mp
ꢀ
1
1734 (C═O α,β-unsaturated furanone), 1621 (C═N).
1
H-NMR (DMSO-d ): δH (ppm) 8.90 (s, 1H, CH═N),
2
-(-2-(((1,3-Diphenyl-1H-pyrazol-4-yl)methylene)
6
hydrazono)-4-oxothiazolidin-5-yl) acetic acid (7).
crystals, mp 260–262°C, [Lit. [18] mp 236–238°C].
White
8.41 (s, 1H, C─H pyrazolyl), 7.92–7.38 (m, 10H,
Ar─H), 7.34 (s, 1H, CH═). MS (m/z, %): 399 (M , 18).
+.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

S. K. Ramadan and H. A. Sallam
Vol 000
Anal. Calcd for C H N O S (399.43): C, 63.15; H, 3.28;
1,2-Bis((1,3-diphenyl-1H-pyrazol-4-yl)methylene)
2
1 13 5 2
hydrazine (17).
White crystals, mp 222–224°C, yield
N, 17.53; S, 8.03. Found: C, 63.11; H, 3.22; N, 17.61; S,
.98.
7
1%. [Lit. [8] mp 220–222°C].
7
Reaction of 2 with malonic acid. A mixture of 2 (2 mmol)
Ring closure of thiosemicarbazone 2 using pyridine.
A
and malonic acid (2 mmol) in acetylchloride (5 mL) was
heated on water bath at 55°C for 4 h. The reaction mixture
was then cooled and poured onto ice-cold water with
stirring. The yellow precipitated solid was collected
by filtration, dried, and then recrystallized from
ethanol/dioxane mixture (2:1) to afford pyrimidinone 18.
solution of 2 (2 mmol) in pyridine (10 mL) was heated
under reflux for 6 h and then left to cool at room
temperature. The obtained precipitate was collected by
filtration and recrystallized from dioxane to afford
mercaptotriazole derivative 14.
5-(1,3-Diphenyl-1H-pyrazol-4-yl)-2,4-dihydro-3H-1,2,4-
3
-(((1,3-Diphenyl-1H-pyrazol-4-yl)methylene)amino)-6-
triazole-3-thione (14).
yield 59%. IR (KBr) (ν, cm ): 3390 (NH), 3052
White crystals, mp 246–248°C,
ꢀ
1
oxo-2-thioxo-1,2,3,6-tetrahydro-pyrimidin-4-yl acetate
(
(
18).
Yellow crystals, mp 338–340°C, yield 68%. IR
1
(
aryl-H), 1617 (C═N), 1213 (C═S). H-NMR (DMSO-
d6): (thiolactam and thiolactim isomers, 1:1) δH (ppm)
.30 (s, 1H, NH, D O-exchangeable), 9.12 and 8.67 (two
ꢀ
1
KBr) (ν, cm ): 3421 (OH, lactim form), 3257 (NH,
lactam form), 3061 (aryl-H), 2978 (alkyl-H), 1721 (C═O
9
2
ester), 1672 (C═O pyrimdinone), 1600 (C═N), 1213
singlets, 1H, C─H pyrazolyl), 8.85 (1H, SH, D O-
2
1
(
C═S). H-NMR (DMSO-d ): (E-isomer and Z-isomer,
6
exchangeable), 8.55 (s, 1H, NH, D O-exchangeable),
2
1
:1) δH (ppm) 12.86 (br. s, 1H, OH, lactim form, D O-
2
8
2
.02–7.99 (d, 2H, Ph─C═N, J = 8.1 Hz), 7.87–7.84 (d,
H, Ph─N, J = 8.4 Hz) 7.82–7.36 (m, 6H, Ar─H). MS
exchangeable), 10.84 and 10.79 (two singlets, 1H, NH,
D O-exchangeable), 9.88 and 9.77 (two singlets, 1H,
+
.
2
(
(
m/z, %): 319 (M , 42). Anal. Calcd for C H N S
319.39): C, 63.93; H, 4.10; N, 21.93; S, 10.04. Found:
17 13 5
CH═N), 8.31 and 8.26 (two singlets, 1H, C─H
pyrazolyl), 7.97–7.49 (m, 10H, Ar─H), 7.36 (s, 1H,
C, 63.81; H, 3.98; N, 21.85; S, 9.98.
C─H pyrimidine), 2.03 and 1.97 (two singlets, 3H, CH ).
3
Ring closure of thiosemicarbazone 2 using HCl/AcOH.
Thiosemicarbazone 2 (2 mmol) was added to equimolar
+.
MS (m/z, %): 431 (M , 25). Anal. Calcd for
C H N O S (431.47): C, 61.24; H, 3.97; N, 16.23; S,
22 17 5 3
mixture of HCl/CH COOH (10 mL), and then the
3
7
.43. Found: C, 61.15; H, 3.81; N, 16.18; S, 7.34.
reaction mixture was heated under reflux 4 h. The
reaction mixture was cooled and poured onto ice-cooled
water. The precipitated solid was collected by filtration
and recrystallized from ethanol to afford the fused
heterocycle derivative 15.
General procedure for the reaction of 2 with acetylacetone,
ethyl benzoylacetate, and/or arylidene malononitrile.
A
solution of thiosemicarbazone
acetylacetone, ethyl benzoylacetate,
2
(2 mmol) and
or 2-(4-
methoxybenzylidene) malononitrile (2 mmol) in absolute
ethanol (20 mL) was refluxed in the presence of sodium
ethoxide (0.05 g Na in 20-mL ethanol) for 8–10 h. The
whole mixture was then cooled and poured onto ice/HCl.
The precipitated solid was filtered off, dried, and then
recrystallized from ethanol to furnish pyrimidine
derivatives 19–21, respectively.
1
,3-Diphenyl-1,6-dihydropyrazolo[3,4-c]pyrazole (15).
Greenish yellow crystals, mp 238–240°C, yield 62%.
ꢀ
1
IR (KBr) (ν, cm ): 3343 (br., NH), 3064 (aryl-H),
1
1
1
7
618 (C═N). H-NMR (DMSO-d ): δ (ppm) 13.20 (s,
6 H
H, NH, exchangeable with D O), 8.40 (CH═N),
.90–7.42 (m, 10H, two phenyl). MS (m/z, %): 260
M , 13). Anal. Calcd for C H N (260.30): C,
2
+.
(
16 12 4
1
-(((1,3-Diphenyl-1H-pyrazol-4-yl)methylene)amino)-4,6-
7
2
3.83; H, 4.65; N, 21.52. Found: C, 73.71; H, 4.52; N,
1.61.
dimethylpyrimidine-2(1H)-thione (19).
crystals, mp 241–243°C, yield 52%. IR (KBr) (ν, cm ):
Faint brown
ꢀ
1
Hydrazinolysis of thiosemicarbazone 2.
hydrate (2.2 mmol, 80%) was added to solution of
Hydrazine
3056 (aryl-H), 2972 (alkyl-H), 1619 (C═N), 1220 (C═S).
1
H-NMR (DMSO-d ): δ (ppm) 8.90 (s, 1H, CH═N),
6 H
thiosemicarbazone 2 (2 mmol) in absolute ethanol
8.40 (s, 1H, C─H pyrazolyl), 7.81–7.45 (m, 10H, Ar─H),
7.02 (s, 1H, C─H pyrimidine), 2.25 and 2.22 (two
(20 mL) and heated under reflux for 6 h. The reaction
+.
mixture was concentrated and then left to cool at room
temperature. The obtained colorless crystals were
collected by filtration and found to be thiosemicarbazide
singlets, 6H, two CH ). MS (m/z, %): 385 (M , 33). Anal.
Calcd for C22H N S (385.49): C, 68.55; H, 4.97; N,
19 5
3
18.17; S, 8.32. Found: C, 68.50; H, 4.94; N, 18.20; S, 8.27.
[mp, mixed mp, TLC, IR]. The residual part was
1-(((1,3-Diphenyl-1H-pyrazol-4-yl)methylene)amino)-6-
recrystallized from benzene/ethanol mixture (1:1) and
found to be diheterylhydrazine 17. The latter compound
was identical in all respects (IR, mp, mixed mp, and
TLC) with an authentic sample prepared by the
condensation of 1,3-diphenylpyrazole-4-carboxaldehyde 1
with hydrazine hydrate in ethanolic solution [8].
hydroxy-4-phenylpyrimidine-2(1H)-thione (20).
crystals, mp 236–238°C, yield 59%. IR (KBr) (ν, cm ):
Beige
ꢀ
1
3444 (br. OH), 3062 (aryl-H), 1618 (C═N), 1261 (C═S).
1
H-NMR (DMSO-d
O-exchangeable), 9.18 (s, 1H, CH═N), 8.20 (s, 1H,
C─H pyrazolyl), 7.85–7.37 (m, 15H, Ar─H), 7.01 (s,
): δ (ppm) 11.20 (br. s, 1H, OH,
6
H
D
2
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2018
Synthesis, Spectral Characterization, Cytotoxic, and Antimicrobial Activities
+
.
1
H, C─H pyrimidine). MS (m/z, %): 449 (M , 52). Anal.
(s, 1H, CH═N), 8.34 (s, 1H, C─H pyrazolyl), 8.20 (s,
1H, C─H quinoline), 7.88–7.71 (m, 4H, quinoline),
7.68–7.38 (m, 10H, two phenyl). MS (m/z, %): 476 (M ,
Calcd for C H N OS (449.53): C, 69.47; H, 4.26; N,
5.58; S, 7.13. Found: C, 69.42; H, 4.18; N, 15.61; S, 7.08.
26 19 5
+.
1
1
5). Anal. Calcd for C H N OS (476.56): C, 68.05; H,
6
-Amino-1-(((1,3-diphenyl-1H-pyrazol-4-yl)methylene)
amino)-4-(4-methoxyphenyl)-2-thioxo-1,2-dihydropyrimidine-
-carbonitrile (21). Faint brown crystals, mp 270–272°C,
27 20 6
4.23; N, 17.64; S, 6.73. Found: C, 68.01; H, 4.17; N,
17.70; S, 6.68.
5
ꢀ
1
yield 44%. IR (KBr) (ν, cm ): 3326, 3256 (NH ), 3062
aryl-H), 2971 (alkyl-H), 2216 (C≡N), 1618 (C═N), 1220
C═S). H-NMR (DMSO-d ): δ (ppm) 6.50 (s, 2H, NH ,
2
Reaction of 2 with chromone-3-carboxaldehyde.
A
(
(
solution of thiosemicarbazone 2 (2 mmol), chromone-3-
carboxaldehyde (2 mmol) in absolute ethanol (20 mL)
was heated under reflux for 5 h. The precipitated solid
while heating was collected by filtration and
recrystallized from dioxane to give the chromone
derivative 24.
1
6
H
2
D O-exchangeable), 9.20 (s, 1H, CH═N), 8.31 (s, 1H,
2
C─H pyrazolyl), 7.91–7.88 (d, 2H, p-anisyl, J = 8.8 Hz),
.83–7.81 (d, 2H, Ph─C═N, J = 8.6 Hz), 7.68–7.66 (d,
H, Ph─N, J = 8.5 Hz), 7.03–7.01 (d, 2H, p-anisyl,
7
2
J = 8.8 Hz), 7.55–7.36 (m, 6H, Ar─H), 3.75 (s, 3H,
2
-((1,3-Diphenyl-1H-pyrazol-4-yl)methylene)-N-(3-formyl-
+.
CH ). MS (m/z, %): 503 (M , 23). Anal. Calcd for
3
4-oxo-4H-chromen-2-yl)-hydrazine-1-carbothioamide (24).
C H N OS (503.58): C, 66.78; H, 4.20; N, 19.47; S,
2
8
21
7
Brown crystals, mp 234–236°C, yield 69%. IR (KBr) (ν,
cm ): 3427, 3331, 3258 (2 NH, H-bonding), 3059 (aryl-
ꢀ
1
6.37. Found: C, 66.72; H, 4.15; N, 19.51; S, 6.32.
Condensation of
p-anisaldehyde.
2
with ethyl cyanoacetate and
H), 1680 (C═O aldehyde), 1648 (C═O chromone), 1619
1
Equimolar mixture of 2, ethyl
(C═N), 1219 (C═S). H-NMR (DMSO-d ): δH (ppm)
6
cyanoacetate, and p-anisaldehyde (2 mmol) in absolute
ethanol (20 mL) was treated with sodium ethoxide (0.05 g
Na in 20-mL ethanol). The whole mixture was heated
under reflux for 6 h. The obtained solid during heating
was filtered off and recrystallized from ethanol to afford
hydroxypyrimidine derivative 22.
11.35 (s, 1H, NH, D O-exchangeable), 10.81 (s, 1H, NH,
2
D O-exchangeable), 9.95 (s, 1H, CHO), 8.62 (s, 1H,
2
CH═N), 8.41 (s, 1H, C─H pyrazolyl), 7.95–7.67 (m, 4H,
chromone), 7.63–7.38 (m, 10H, two phenyl). MS (m/z,
+.
%): 493 (M , 33). Anal. Calcd for C H N O S
2
7 19 5 3
(493.54): C, 65.71; H, 3.88; N, 14.19; S, 6.50. Found: C,
65.59; H, 3.73; N, 14.27; S, 6.39.
1
-(((1,3-Diphenyl-1H-pyrazol-4-yl)methylene)amino)-6-
hydroxy-4-(4-methoxyphenyl)-2-thioxo-1,2-
dihydropyrimidine-5-carbonitrile (22).
Biological assay. Antimicrobial activity.
methods.
Materials and
Antimicrobial activity of the tested samples
Beige crystals,
ꢀ
1
mp 254–256°C, yield 67%. IR (KBr) (ν, cm ): 3445
br., OH), 3063 (aryl-H), 2972 (alkyl-H), 2215 (C≡N),
was determined using a modified Kirby–Bauer disc
diffusion method [20]. Briefly, 100 μL of the test
bacteria/fungi were grown in 10 mL of fresh media until
they reached a count of 108 cells/mL for bacteria or 105
cells/mL for fungi approximately [22]. A 100 μL of
microbial suspension was spread onto agar plates
corresponding to the broth in which they were maintained.
Isolated colonies of each organism that might be playing
a pathogenic role should be selected from primary agar
plates and tested for susceptibility by disc diffusion
method [23]. Of the many media available, National
Committee for Clinical Laboratory Standards recommends
Mueller–Hinton agar due to its results in good batch-to-
batch reproducibility. Disc diffusion method for
filamentous fungi tested by using approved standard
method (M38-A) developed [24] for evaluating the
susceptibilities of filamentous fungi to antifungal agents.
Disc diffusion method for yeasts developed by using
approved standard method (M44-P) [25]. Plates inoculated
with filamentous fungi as Aspergillus flavus at 25°C for
48 h; Gram (+) bacteria as Staphylococcus aureus,
Bacillus subtilis; Gram (ꢀ) bacteria as Escherichia coli,
Pseudomonas aeuroginosa they were incubated at
35–37°C for 24–48 h and yeast as Candida albicans
incubated at 30°C for 24–48 h and then the diameters of
the inhibition zones were measured in millimeters [21].
(
1
1
627 (C═N), 1262 (C═S). H-NMR (DMSO-d ): δH
6
(
(
(
ppm) 11.31 (br. s, 1H, OH, D O-exchangeable), 9.17
s, 1H, CH═N), 8.21 (s, 1H, C─H pyrazolyl), 7.90–7.87
d, 2H, p-anisyl, J = 8.8 Hz), 7.84–7.82 (d, 2H,
2
Ph─C═N, J = 8.7 Hz), 7.67–7.65 (d, 2H, Ph─N,
J = 8.5 Hz), 7.03–7.01 (d, 2H, p-anisyl, J = 8.8 Hz),
7
%
.56–7.37 (m, 6H, Ar─H), 3.79 (s, 3H, CH ). MS (m/z,
): 504 (M , 41). Anal. Calcd for C H N O S
3
+
.
28 20 6 2
(504.57): C, 66.65; H, 4.00; N, 16.66; S, 6.35. Found: C,
66.61; H, 3.95; N, 16.69; S, 6.30.
Reaction of 2 with 2-chloroquinoline-3-carboxaldehyde.
-Chloroquinoline-3-carboxaldehyde (2 mmol) was treated
2
with a solution of thiosemicarbazone 2 (2 mmol) in glacial
acetic acid (10 mL). The reaction mixture was heated under
reflux for 8 h. The yellow precipitated solid after cooling
was filtered off and recrystallized from dioxane to give
the quinoline derivative 23.
2
-((1,3-Diphenyl-1H-pyrazol-4-yl)methylene)-N-(3-
formylquinolin-2-yl)hydrazine-1-carbothioamide (23).
Yellow crystals, mp 263–265°C, yield 52%. IR (KBr) (ν,
ꢀ
1
cm ): 3415 (br., NH, H-bonding), 3057 (aryl-H), 1662
1
(
C═O), 1622 (C═N), 1216 (C═S). H-NMR (DMSO-d ):
6
δH (ppm) 11.30 (s, 1H, NH, D O-exchangeable), 10.50
2
(
s, 1H, NH, D O-exchangeable), 9.75 (s, 1H, CHO), 8.55
2
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

S. K. Ramadan and H. A. Sallam
Vol 000
Standard discs of Ampicillin (antibacterial agent),
Amphotericin B (antifungal agent) served as positive
controls for antimicrobial activity but filter discs
impregnated with 10 μL of solvent (distilled water,
chloroform, DMSO) were used as a negative control. The
agar used is Meuller–Hinton agar that is rigorously tested
for composition and pH. Further, the depth of the agar in
the plate is a factor to be considered in the disc diffusion
method. This method is well documented, and standard
zones of inhibition have been determined for susceptible
and resistant values.
After incubation, the cells treated with different
concentrations of compounds and incubated for 24 h.
After 24 h of drug treatment, 20 μl of MTT solution at
5 mg/ml was added and then incubated for 4 h. DMSO in
volume of 100 μL is added into each well to dissolve the
purple formazan formed. The colorimetric assay is
measured and recorded at absorbance of 570 nm using a
plate reader (EXL 800, Bio-Tech, Winoosky, VT, USA).
The relative cell viability in percentage was calculated
as (A570 of treated samples/A570 of untreated
sample) × 100.
Blank paper disks (Schleicher and Schuell, Spain) with
a diameter of 8.0 mm were impregnated 10 μL of tested
concentration of the stock solutions. When a filter paper
disc impregnated with a tested chemical is placed on agar,
the chemical will diffuse from the disc into the agar. This
diffusion will place the chemical in the agar only around
the disc. The solubility of the chemical and its molecular
size will determine the size of the area of chemical
infiltration around the disc. If an organism is placed on
the agar, it will not grow in the area around the disc if
it is susceptible to the chemical. This area of no growth
around the disc is known as “Zone of inhibition” or
REFERENCES AND NOTES
[
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