Thiazolidin-4-one, azetidin-2-one and 1,3,4-oxadiazole derivatives of isonicotinic acid hydrazide: Synthesis and their biological evaluation

Article abstract of DOI:10.2298/JSC101104092G

A series of thiazolidin-4-one (2a-h; 3a-h), azetidin-2-one (4a-h) and 1,3,4-oxadiazole (5a-h) derivatives of isoninicotinic acid hydrazide (INH) were synthesized in order to obtain new compounds with potential anti-inflammatory, analgesic, ulcerogenic and

Full text of DOI:10.2298/JSC101104092G

J. Serb. Chem. Soc. 76 (8) 1057–1067 (2011)
JSCS–4184
UDC 547.298.61+66.094.3–92:541.459+
547.915.5:615.212:615.276
Original scientific paper
Thiazolidin-4-one, azetidin-2-one and 1,3,4-oxadiazole
derivatives of isonicotinic acid hydrazide: synthesis
and their biological evaluation
1
1
2
SADAF J. GILANI1,2 , SUROOR A. KHAN , OZAIR ALAM , VIJENDER SINGH
*
and ALKA ARORA2
1Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Jamia Hamdard,
New Delhi-110062 and 2Department of Pharmaceutical Chemistry,
KIET School of Pharmacy, Ghaziabad, U.P. 201206, India
(Received 4 November 2009, revised 2 April 2011)
Abstract: A series of thiazolidin-4-one (2a–h; 3a–h), azetidin-2-one (4a–h) and
1,3,4-oxadiazole (5a–h) derivatives of isoninicotinic acid hydrazide (INH)
were synthesized in order to obtain new compounds with potential anti-inflam-
matory, analgesic, ulcerogenic and lipid peroxidation activities. The structures
of the new compounds were supported by their IR, 1H-NMR and mass spectral
data. All compounds were evaluated for their anti-inflammatory activity by the
carrageenan-induced rat paw edema test method. Eleven of the new com-
pounds, out of 32, showed very good anti-inflammatory activity in the carra-
geenan-induced rat paw edema test, with significant analgesic activity in the
tail immersion method together with negligible ulcerogenic action. The com-
pounds, which showed less ulcerogenic action, also showed reduced malondial-
dehyde content (MDA), which is one of the by-products of lipid peroxidation.
The study showed that the compounds inhibited the induction of gastric mu-
cosal lesions and it can be suggested from the results that their protective ef-
fects may be related to inhibition of lipid peroxidation in the gastric mucosa.
Keywords: anti-inflammatory activity; analgesic activity; ulcerogenic activity;
lipid peroxidation.
INTRODUCTION
The currently available non-steroidal anti-inflammatory drugs (NSAIDs),
such as ibuprofen, flurbiprofen, fenbufen and naproxen, exhibit gastric toxicity.
Long-term use of these drugs has been associated with gastro-intestinal (GI) ul-
1
ceration, bleeding and nephrotoxicity. The pharmacological activity of NSAIDs
is related to the suppression of prostaglandin biosynthesis from arachidonic acid
*Corresponding author. E-mail: gilanisadaf@gmail.com
doi: 10.2298/JSC101104092G
1057
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GILANI et al.
2,3
by inhibiting the enzyme cyclooxygenases (COXs). Recently, it was disco-
vered that COX exists in two isoforms, COX-1 and COX-2, which are regulated
4–6
differently.
COX-1 provides cytoprotection in the gastrointestinal (GI) tract,
7–9
whereas inducible COX-2 mediates inflammation.
on the market show greater selectivity for COX-1 than COX-2, chronic use of
NSAIDs may elicit appreciable GI irritation, bleeding and ulceration.
Since most of the NSAIDs
10
The GI damage from NSAIDs is generally attributed to two factors, i.e., lo-
cal irritation by the carboxylic acid moiety common to most NSAIDs (topical ef-
fect); and decreased tissue prostaglandin production, which undermines the phy-
siological role of the cytoprotective prostaglandins in maintaining GI health and
11
homeostasis. The incidence of clinically significant GI side effects due to
12
NSAIDs is high (30 %) and causes some patients to abandon NSAID therapy.
Thus, the discovery of COX-2 provided the rationale for the development of drugs
devoid of GI disorders while retaining clinical efficacy as anti-inflammatory
agents. However, recent reports showed that selective COX-2 inhibitors (coxibs)
13
could lead to adverse cardiovascular effects. Therefore, the development of no-
vel compounds having anti-inflammatory and analgesic activity with improved
safety profiles is still a necessity.
Synthetic approaches based on chemical modification of NSAIDs have been
undertaken with the aim of improving their safety profiles. A literature survey
revealed that derivatization of the carboxylate function of NSAIDs resulted in re-
14–17
tained anti-inflammatory activity with reduced ulcerogenic potential.
It has
also been reported in the literature that certain compounds bearing a thiazolidin-
-4-one, azetidin-2-one or 1,3,4-oxadiazole nucleus possess significant anti-in-
18–20
flammatory activity.
In view of the important biological properties of the thiazolidin-4-one, aze-
tidin-2-one and 1,3,4-oxadiazole nuclei, it was planned to suitably incorporate
the above-mentioned ring systems into the isoniazide moiety to explore the pos-
sibilities of some altered biological actions. Hence, the thiazolidin-4-one, azeti-
din-2-one and 1,3,4-oxadiazole derivatives reported herein were designed and
synthesized. They were found to possess an interesting profile of anti-inflam-
matory and analgesic activity with significant reduction in their ulcerogenic risks
to the stomach.
EXPERIMENTAL
Materials, methods and instrumentation
All the solvents were of AR grade and were obtained from Merck, CDH and S.D. Fine
chemicals. Melting points were determined in open capillary tubes and are uncorrected. All
the compounds were subjected to elemental analysis (CHN) and the measured values agreed
within ±0.4 % with the calculated ones. Thin layer chromatography was performed on silica
gel G (Merck). The spots were developed in an iodine chamber and visualized with an ultra-
violet lamp. The solvent systems used were benzene:acetone (8:2, v/v) and toluene:ethyl ace-
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NOVEL ISONICOTINIC ACID HYDRAZIDE DERIVATIVES
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tate:formic acid (5:4:1, v/v). Ashless Whatman No. 1 filter paper was used for vacuum fil-
tration. The IR spectra were recorded in KBr pellets on a (BIO-RAD FTS 135) WIN-IR spec-
trophotometer. The FAB mass spectra of all the compounds were recorded on a JEOL SX102/
/DA-600 mass spectrometer using argon/xenon (6 kV, 10 mA) as the FAB gas. The 1H-NMR
spectra were recorded on a Bruker model DPX 300 FT-NMR spectrometer in CDCl3 using
tetramethylsilane (Me4Si, TMS) as an internal standard. The chemical shifts are reported in
the δ ppm scale.
General procedure for the preparation of the new (E)-N’-(substituted benzylidene)-
isonicotinohydrazides (1a–h)
To an equimolar methanolic solution of isonicotinic acid hydrazide (0.1 mol) and sub-
stituted benzaldehyde (0.1 mol), a few drops of glacial acetic acid were added. The mixture
was then refluxed on water bath for 5–6 h. It was then allowed to cool, poured onto crushed
ice and recrystallised from methanol.
General procedure for the preparation of the new N-(2-(substituted phenyl)-4-oxothiazolidin-
-3-yl)isonicotinamides (2a–h)
A mixture of 1 (0.01 mol) and thioglycolic acid (0.01 mol) was heated on an oil-bath at
120–125 °C for 12 h. The reaction mixture was cooled and treated with 10 % sodium bi-
carbonate solution. The product was isolated and recrystallised from methanol–dioxane (4:1).
General procedure for the preparation of new 2-(2-(2-substituted phenyl)-3-(isonicotin-
amido)-4-oxothiazolidin-5-yl)acetic acids (3a–h)
A mixture of 1 (0.01 mol) and thiomalic acid (0.01 mol) was heated on an oil-bath at
120–125 °C for 12 h. The reaction mixture was cooled and treated with 10 % sodium bi-
carbonate solution. The product was isolated and recrystallised from methanol–dioxane (4:1).
General procedure for the synthesis of N-(3-chloro-2-(2-substituted phenyl)-4-oxoazetidin-1-
-yl)isonicotinamide (4a–h)
A solution of 1 (0.01 mol) in dioxane (20 mL) was added to a well-stirred mixture of
chloroacetyl chloride (0.012 mol) and triethylamine (Et3N) (0.012 mol) in dioxane (10 mL) at
0–5 °C. The reaction mixture was then stirred for 8 h, kept for 2 days at room temperature and
then treated with cold water. The solid thus obtained was filtered, washed with water and re-
crystallised from methanol.
General procedure for the synthesis of 1-(2-(2-substituted phenyl)-5-(pyridin-4-yl)-1,3,4-
-oxadiazol-3(2H)-yl)ethanone (5a–h)
A mixture of 1 (0.003 mol) and acetic anhydride (10 mL) was heated under reflux for 4
h. After the reaction mixture attained room temperature, the excess acetic anhydride was
decomposed by water and the mixture was stirred for further 30 min. The separated product
was filtered, washed with water, dried and recrystallised from appropriate solvent systems.
Biological evaluation
Animals. Adult Wistar strain rats of either sex weighing 150–200 g were used for the
evaluation of the anti-inflammatory, ulcerogenic and lipid peroxidation activities, whereas
Swiss albino mice weighing 25–30 g were used for assessing the analgesic activity. The ani-
mals were allowed food and water ad libitum except during the experiments. They were
housed in a room at 25±2 °C, and 50±5 % relative humidity with a 12 h light/dark cycle. The
animals were randomly allocated into groups at the beginning of all the experiments. All the
experimental protocols were performed with permission from the Institutional Animal Ethics
Committee (IAEC), form No. 520. The animals were obtained from the Central Animal House
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Facility, Hamdard University, New Delhi-110062, India. Registration number and date of re-
gistration of the Animal House Facility (173/CPCSEA, 28, JAN-2000). All the test com-
pounds and the reference drug were administered orally suspended in 0.5 % carboxymethyl
cellulose (CMC) solutions.
Anti-inflammatory activity
The synthesized compounds were evaluated for their anti-inflammatory activity using the
carrageenan-induced hind paw edema method.21 The animals were randomly allocated into
groups of six animals each and were fasted for 24 h before the experiment with free access to
water. The control group received only 0.5 % carboxymethyl cellulose solution. The standard
drug naproxen was administered orally at a dose of 10 mg kg-1. The test compounds were
administered orally at an equimolar oral dose relative to 10 mg kg-1 naproxen. Into the sub-
-plantar region of the right hind paw of each rat, 0.1 ml of 1 % carrageenan solution in saline
was injected subcutaneously, 1 h after the administration of the test compounds or the stan-
dard drug. The right hind paw volume was measured before and after 3 and 4 h of carrageenan
treatment by means of a plethysmometer. The percent edema inhibition was calculated from
the mean effect in the control and treated animals according to the following equation:
Percent edema inhibition = (Vc – Vt /Vc)×100
where Vt represents the mean increase in paw volume in rats treated with test compounds and
Vc represents the mean increase in paw volume in the control group of rats.
Analgesic activity
The analgesic activity was evaluated by the tail immersion method.22 Swiss albino mice
allocated into different groups consisting of six animals of either sex in each, weighing 25–30
g, were used for the experiment. The analgesic activity was evaluated after oral administration
of the test compounds at an equimolar dose relative to 10 mg kg-1 naproxen. The test com-
pounds and the standard drug were administered orally as a suspension in carboxymethyl cel-
lulose solution in water (0.5 %, w/v). The analgesic activity was assessed before and 4 h after
the administration of test compounds and standard drug. The lower 5 cm portion of the tail
was gently immersed into thermostatically controlled water at 55±0.5 °C. The time in seconds
for tail withdrawal from the water was taken as the reaction time with a cut-off time of im-
mersion set at 10 s for both the control as well as the treated group of animals.
Acute ulcerogenicity
Acute ulcerogenicity was determined according to the method of Cioli et al.23 The ani-
mals were allocated into different groups consisting of six animals in each group. The ulcero-
genic activity was evaluated after oral administration of the test compounds at an equimolar
dose relative to 30 mg kg-1 naproxen. The control group received only 0.5 % carboxymethyl
cellulose solution. Food but not water was removed 24 h before administration of the test
compounds. After the drug treatment, the rats were fed with a normal diet for 17 h and then
sacrificed. The stomach was removed and opened along the greater curvature, washed with
distilled water and cleaned gently by dipping in normal saline. The mucosal damage was exa-
mined by means of a magnifying glass. For each stomach the mucosal damage was assessed
according to the following scoring system: 0.5, redness, 1.0, spot ulcers, 1.5, hemorrhagic
streaks, 2.0, ulcers < 3, 3.0, ulcers > 3 but ≤ 5. The mean score of each treated group minus
the mean score of control group was regarded as the severity index of gastric mucosal damage.
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NOVEL ISONICOTINIC ACID HYDRAZIDE DERIVATIVES
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Lipid peroxidation
Lipid peroxidation in the gastric mucosa was determined according to the method of
Ohkawa et al.24 After screening for ulcerogenic activity, the gastric mucosa was scraped with
two glass slides, weighed (100 mg) and homogenized in 1.8 ml of 1.15 % ice-cold KCl solu-
tion. The homogenate was supplemented with 0.2 ml of 8.1 % sodium dodecyl sulphate (SDS),
1.5 ml of acetate buffer (pH 3.5) and 1.5 ml of 0.8 % thiobarbituric acid (TBA). The mixture
was heated at 95 °C for 60 min. After cooling, the reactants were supplemented with 5 ml of
the mixture of n-butanol and pyridine (15:1 v/v), shaken vigorously for 1 min and centrifuged
for 10 min at 4000 rpm. The supernatant organic layer was removed and the absorbance
measured at 532 nm using a UV/Vis spectrophotometer. The results were expressed as nmol
MDA 100 mg-1 tissue, using an extinction coefficient 1.56×105 cm-1 M-1.
Statistical analysis of data
The data are expressed as mean±standard error of mean (SEM). In the anti-inflammatory,
ulcerogenic and lipid peroxidation studies, statistical differences between the treatments and
the standard were tested by one-way ANOVA followed by the Dunnett multiple comparison
test. A value of p < 0.01 was considered significant. In analgesic activity study, the statistical
differences in treatments and standard were tested by the paired Student’s t-test.
RESULTS AND DISCUSSION
Chemistry
The key intermediates used in the synthesis of thiazolidin-4-ones 2a–h and
3a–h, azetidin-2-one 4a–h and 1,3,4-oxadiazole derivatives 5a–h, (E)-N’-(2-sub-
stituted benzylidene)isonicotinohydrazides 1a–h were prepared starting from iso-
nicotinic acid hydrazide. The reaction of isonicotinic acid hydrazide with sub-
stituted benzaldehyde in refluxing methanol with few drops of glacial acetic acid
gave the (E)-N’-(2-substituted benzylidene)isonicotinohydrazides 1a–h. In the
present study, the reaction of the substituted benzylidene isonicotinohydrazides
1a–h with thioglycolic acid, thiomalic acid, chloroacetyl chloride and acetic an-
hydride in presence of various reagents gave the new thiazolidin-4-ones 2a–h and
3a–h, azetidin-2-one 4a–h and 1,3,4-oxadiazole derivatives 5a–h, respectively.
The synthesis routes to the compounds are outlined in Scheme 1. The ana-
lytical and spectral data of the newly prepared compounds are given in the sup-
plementary material to this paper.
The IR spectra of compounds 1a–h showed absorption peaks at 3300, 1680
–1
and 1600 cm due to N–H, C=O and –N=CH stretching vibrations. The appea-
rance of the stretching of the C=O of thiazolidinone, β-lactam ring and the acetyl
–1
of the oxadiazole ring at 1700, 1745 and 1660 cm , respectively, in the spectra
–1
of the derivatives, together with the C=O stretching at 1680 cm confirmed the
formation of the compounds 2a–h, 3a–h, 4a–h and 5a–h.
1
The H-NMR spectra of compounds 1a–h revealed a multiplet at δ 7.72 and
8.63 ppm for pyridine and δ 7.12–7.15 ppm for the aromatic ring and singlets at δ
6.1 and 7.9 ppm for –NH and –N=CH, respectively. The disappearance of the
singlet peak of –N=CH and presence of a singlet peak at δ 3.5, 10.0, 7.7 and 7.19
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GILANI et al.
ppm of –CH of thiazolidinone, COOH, –N–CH and –CH oxadiazole, respecti-
2
vely, proved that these compounds participated in the cyclisation reaction and
formed the desired compounds.
Scheme 1.The synthesis routes to the title compounds.
Anti-inflammatory activity
The anti-inflammatory activities of the synthesized compounds 2a–h, 3a–h,
4a–h and 5a–h were evaluated by the carrageenan-induced paw edema method
15
–1
of Winter et al. The compounds were tested at a 10 mg kg oral dose and were
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NOVEL ISONICOTINIC ACID HYDRAZIDE DERIVATIVES
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compared with the standard drug naproxen at the same oral dose. The tested
compounds showed anti-inflammatory activity ranging from 31.81–84.54 % (Ta-
ble I), whereas the standard drug naproxen showed 81.86 % inhibition after 4 h.
TABLE I. Biological data of isonicotinic aid hydrazide derivatives (anti-inflammatory and
analgesic activities of the test compounds with reference to the control. Ulcerogenic and lipid
peroxidation were compared with reference to the standard drug, i.e., naproxen. Data were
only analyzed by the Student’s t-test for n = 6)
(nmol MDA
Anti-inflam-
matoryactivity
(% inhibition±SEM) hibition±SEM) index±SEM)
Analgesic
activity (% in- tivity (severity
Ulcerogenic ac-
content±
SEM)/100 mg
tissue
Compd.
R
Control
Naproxen
2a
2b
2c
2d
2e
2f
2g
2h
3a
3b
3c
3d
3e
3f
3g
3h
4a
4b
4c
4d
4e
4f
4g
4h
5a
–
–
–
–
0.000±0.00
2.250±0.11
3.25±0.005
9.04±0.24
81.86±3.71c
78.00±2.53c
64.64±3.79c
57.57±3.45c
34.08±3.47c
39.54±2.27c
82.81±2.17a
84.24±0.95a
65.15±1.51c
77.27±1.92c
63.63±1.96c
68.93±3.60c
31.81±3.45c
56.81±1.94c
75.00±2.53c
82.27±1.84a
59.09±3.60c
70.45±3.26c
68.93±3.60c
84.54±1.94a
44.05±3.76c
54.28±3.45c
77.14±2.20c
76.50±1.82c
72.72±1.66c
82.86±1.84a
69.23±3.71c
63.46±2.38c
51.28±3.61c
31.81±3.41c
76.92±1.58b
83.80±1.85a
68.26±2.31b
71.42±1.31a
70.32±1.31a
59.09±3.60c
65.15±1.51c
44.05±3.76c
31.81±3.71c
71.57±1.85a
72.42±1.31a
59.23±3.71c
70.45±3.26a
54.28±3.76c
62.85±1.65c
47.27±2.73c
66.46±2.38c
70.04±1.64a
72.72±1.66a
49.54±2.27c
64.80±1.28c
59.61±1.00c
73.06±2.61a
57.14±2.20c
44.19±2.76c
71.12±1.24a
69.54±3.19c
52.48±3.76c
70.45±3.26a
62.69±1.65c
55.32±1.96c
42.45±3.16c
57.80±2.84b
71.72±1.66a
72.52±1.00a
61.82±1.65b
o-C6H4Cl
p-C6H4Cl
0.583±0.18a 5.715±0.14a
2.750±0.19c 8.808±0.06c
1.000±0.31c 7.904±0.19c
1.667±0.24c 6.715±0.16a
2.140±0.11c 8.155±0.14c
0.666±0.16a 5.608±0.02a
0.510±0.08a 5.876±0.13a
2.427±0.14c 8.128±0.06c
0.833±0.24c 6.110±0.04a
2.466±0.20c 8.742±0.17c
0.917±0.12c 7.788±0.10c
2.835±0.18c 8.110±0.13c
0.814±0.24c 7.337±0.16c
0.541±0.10a 6.854±0.14a
0.516±0.11a 5.788±0.10a
2.413±0.10c 8.371±0.12c
2.683±0.10c 7.874±0.15c
1.751±0.16c 5.192±0.22a
0.517±0.25a 6.651±0.17a
1.836±0.11c 7.327±0.19c
2.432±0.24c 8.418±0.11c
o-C6H4OH
m-C6H4OH
p-C6H4OCH3
p-C6H4F
o-C6H4NO2
p-C6H4N(CH3)2
o-C6H4Cl
p-C6H4Cl
o-C6H4OH
m-C6H4OH
p-C6H4OCH3
p-C6H4F
o-C6H4NO2
p-C6H4N(CH3)2
o-C6H4Cl
p-C6H4Cl
o-C6H4OH
m-C6H4OH
p-C6H4OCH3
p-C6H4F
o-C6H4NO2
p-C6H4N(CH3)2
o-C6H4Cl
0.750±0.18c
5.62±0.29a
0.817±0.12c 8.364±0.24c
1.342±0.23c 7.821±0.14c
0.750±0.14c
7.51±0.68c
5b
5c
5d
5e
5f
p-C6H4Cl
1.967±0.17c 7.441±0.12c
1.834±0.22c 7.652±0.17c
2.667±0.14c 8.345±0.19c
1.684±0.26b 6.791±0.13a
0.667±0.18a 6.338±0.20a
0.412±0.11a 5.742±0.17a
1.327±0.20b 7.155±0.22b
o-C6H4OH
m-C6H4OH
p-C6H4OCH3
p-C6H4F
o-C6H4NO2
p-C6H4N(CH3)2
5g
5h
a
b
c
p < 0.0001; p < 0.001; p < 0.01
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The anti-inflammatory activities of the thiazolidin-4-one derivatives (2a–h
and 3a–h) were in the range 34.08–84.24 %. When the CH of thiazolidin-4-one
2
moiety (2a–h) at the position 5 was substituted by CH COOH (3a–h), no sub-
2
stantial change was observed in the activity. It was observed that thaiazolidin-4-
-one derivatives having o-Cl, p-F and o-NO phenyl group showed better or equi-
2
valent activity (78.00, 82.81, 84.24 (in series 2), 77.27, 75.00 and 82.27 % (in
series 3), respectively) to that of the standard drug. The other thiazolidin-4-one
derivatives showed moderate activity.
The anti-inflammatory activities of the azetidin-2-one derivatives were be-
tween 44.05 and 84.54 %. The highest activity (84.54 %) was found with the aze-
tidin-2-one derivative 4c having an o-hydroxyphenyl group at the position 4. It
was observed that the azetidin-2-one derivative having a p-dimethylaminophenyl
group (4h) also showed good activity, viz. 72.72 % as did those having an
o-chloro, p-fluoro and o-nitrophenyl group (70.45, 77.14 and 76.50 %, respecti-
vely). The other derivatives showed moderate activity.
The 1,3,4-oxadiazole derivatives of isonicotinic acid hydrazide showed anti-
-inflammatory activities ranging from 31.81 to 83.80 %.The highest activity
(83.80 %) of the oxadiazole derivative was found for compound 5g having a
o-nitrophenyl group at the position 2. When this group was replaced by the
o-chloro phenyl group, the activity was found to be decreased but equivalent to
that of the standard drug (82.86 %). Furthermore, the oxadiazole derivative having
a p-fluorophenyl group (5f) at the position 2 also showed good activity (76.92
%). The remaining oxadiazole derivatives showed moderate activity.
Analgesic activity
The thiazolidin-4-one derivatives 2f, 2g and 3g showed analgesic activities
ranging from 71.57 to 72.72 %, i.e., better than that of the standard drug napro-
xen (71.42 %). The presence of a o-nitrophenyl group, at the position 2 of the
thiazolidinone ring (2g and 3g) gave the maximum activity (72.42 and 72.72 %,
respectively). When this group was replaced by o-chlorophenyl (2a and 3a) and
p-fluorophenyl (2f and 3f), the activity was found to be slightly decreased
(70.32–71.57 %). When these groups were replaced by p-OCH and m-OH phe-
3
nyl group (2e and 3d), the activity decreased drastically (31.81 and 47.27 %, res-
pectively). The results show that an electron-withdrawing group increased the
analgesic activity of the compounds.
When the thiazolidin-4-one nucleus was replaced by the azetidin-2-one nuc-
leus, the analgesic activity of most of the compounds decreased, except for com-
pound 4c having an o-hydroxylphenyl group at the position 4, which exhibited
the highest activity (73.06 %). 1,3,4-Oxadiazole derivatives (5a–h) were also
screened and showed 42.45–72.52 % analgesic activity. The compounds having a
p-fluorophenyl (5f) and a o-nitrophenyl group (5g) at the position 2 of the oxa-
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diazole nucleus showed a slight difference in their activity (71.72 and 72.52 %).
There was a decrease in the activity when these groups were replaced by an m-
-hydroxyphenyl group (5d).
Acute ulcerogenesis
The tested compounds showed significant reductions in ulcerogenic activity,
ranging from 0.417±0.08 to 2.835±0.18, whereas the standard drug naproxen ex-
hibited a high severity index (2.250±0.11). The ulcerogenic activity of the thiazo-
lidin-4-one derivatives 2a–h and 3a–h ranged from 0.510±0.08 to 2.835±0.18.
The compounds with a o-nitrophenyl group (2g and 3g) showed minimum ulce-
rogenic activity (0.510±0.08 and 0.516±0.11, respectively). Moreover, their anti-
-inflammatory activity was found to be high viz. 84.24 and 82.27 %, respectively.
The other three thiazolidinone derivatives 2a, 2f and 3f also showed reductions in
ulcerogenic activity (0.583±0.08, 0.666±0.16 and 0.541±0.10, respectively) in
comparison to the standard drug.
The azetidin-2-one derivatives 4c, 4f and 4g also showed reductions in ulce-
rogenic activity (0.517±0.11, 0.750±0.14 and 0.817±0.13). The 1,3,4-oxadiazole
derivatives showed minimal ulcerogenic activity, when compared to the thiazoli-
din-4-one and azetidin-2-one derivatives. Compound 5g having a o-nitrophenyl
group showed the minimum severity index (0.412±0.11).
Lipid peroxidation
It was reported in the literature that compounds showing less ulcerogenic
activity also showed reduced malondialdehyde (MDA) content, a by-product of
25
lipid peroxidation. Therefore, an attempt was made to correlate the decrease in
ulcerogenic activity of the compounds with that of lipid peroxidation. All the
compounds screened for ulcerogenic activity were also analyzed for lipid peroxi-
dation.
The lipid peroxidation was measured as nmol of MDA per 100 mg of tissue.
The naproxen (standard drug) showed the maximum lipid peroxidation (9.04±
±0.24), whereas the control group showed 3.25±0.05. It was found that all the
cyclised derivatives showing lower ulcerogenic activity also exhibited reduced
lipid peroxidation (Table I). Thus, these studies showed that the synthesized
compounds inhibited the induction of gastric mucosal lesions and the results fur-
ther suggested that their protective effect might be related to the inhibition of
lipid peroxidation in the gastric mucosa.
CONCLUSIONS
Various thiazolidin-4-one, azetidin-2-one and 1,3,4-oxadiazole derivatives of
isonicotinic acid hydrazide were prepared with the objective of developing better
anti-inflammatory molecules with minimum ulcerogenic activity. It was interest-
ing to note that six of the cyclised compounds, 2f, 2g, 3g, 4c, 5a and 5g, were
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1066
GILANI et al.
found to have anti-inflammatory activity greater than that of the standard drug
–1
(naproxen, 81.86 %) at 10 mg kg p.o. Furthermore, five compounds, 2a, 3a, 4f,
4g and 5f, exhibited anti-inflammatory activity equivalent to the standard drug in
the carrageenan-induced paw edema test in rats. When these compounds were
subjected to the analgesic activity test, they showed increased activity over the
reference drug.
nd
The presence of p-fluorophenyl and o-nitrophenyl groups at the 2 position
of the thiazolidinone nucleus increased the anti-inflammatory activity whereas
the presence of m-hydroxryphenyl and p-methoxyphenyl groups decreased the
anti-inflammatory activity. It was further noted that the presence of o-hydroxy, o-
-chloro and o-nitrophenyl groups at the and positions 4 and 2 of azetidinone and
oxadiazole nucleus, respectively, increased the anti-inflammatory activity com-
pared to the standard drug, whereas presence of the p-fluorophenyl group showed
anti-inflammatory activity slightly less than that of the standard drug.
These compounds tested for ulcerogenic activity showed a significant de-
crease in activity compared to that of the standard drug. It was noted that the oxa-
diazole derivatives showed maximum reduction in ulcerogenic activity followed
by the thiazolidinone and azetidinone derivatives. It was further concluded that
the presence of p-fluorophenyl and o-nitrophenyl groups at the positions 2, 4 and
2 of thiazolidione, azetidinone and oxadiazole nucleus, respectively, showed maxi-
mum anti-inflammatory and analgesic activity, minimum ulcerogenic activity
together with minimum lipid peroxidation.
SUPPLEMENTARY MATERIAL
Analytical and spectral data of the synthesized compounds are available electronically at
http://www.shd.org.rs/JSCS/, or from the corresponding author on request.
Acknowledgements. The authors are thankful to the Head of the Department of Pharma-
ceutical Chemistry for providing the laboratory facilities and the Central Drug Research
Institute (CDRI) for the spectral analysis of the compounds. The authors are also thankful to
the Animal House, Jamia Hamdard, for providing the Wistar rats and Swiss Albino mice.
И З В О Д
ТИАЗОЛИДИН-4-ОНСКИ, АЗЕТИДИН-2-ОНСКИ И 1,3,4-ОКСАДИАЗОЛСКИ ДЕРИВАТИ
ХИДРАЗИДА ИЗОНИКОТИНСКЕ КИСЕЛИНЕ: СИНТЕЗА И
ЊИХОВА БИОЛОШКА АКТИВНОСТ
SADAF J. GILANI1,2, SUROOR A. KHAN1, OZAIR ALAM1, VIJENDER SINGH2 и ALKA ARORA2
1Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Jamia Hamdard, New Delhi-110062 и
2Department of Pharmaceutical Chemistry, KIET School of Pharmacy, Ghaziabad, U.P. 201206, India
Синтетисана је серија нових тиазолидин-4-онских (2a–h и 3a–h), азетидин-2-oнских
(4a–h) и 1,3,4-оксадиазолских (5a–h) деривата хидразида изоникотинске киселине (INH) са
циљем да се испита њихова анти-инфламаторна, аналгетичка и улцерогена активност и ак-
тивност према пероксидацији липида. Структура синтетисаних једињења одређена је на
основу IC, NMR и масених спектара. Анти-инфламаторна активност једињења одређена је
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NOVEL ISONICOTINIC ACID HYDRAZIDE DERIVATIVES
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тестом карагененом идукованог отока на шапи пацова. Од 32 испитана једињења, 11 пока-
зује врло добру анти-инфламаторну активност и изражену аналгетичку активност, уз
незнатну улцерогену активност. Једињења која показују најмању улцерогену активност до-
воде до снижавања количине малондиалдехида (MDA), који је споредни производ липидне
пероксидације. Резултати показују да једињења инхибирају индукцију оштећења слузокоже
желуца и може се претпоставити да је њихова заштитна улога повезана са инхибицијом
липидне пероксидације у слузокожи желуца.
(Примљено 4. новембра 2009, ревидирано 2. априла 2011)
REFERENCES
1. M. Amir, H. Kumar, S. A. Javed, Eur. J. Med. Chem. 39 (2004) 535
2. C. J. Smith, Y. Zhang, C. M. Koboldt, J. Muhammad, B. S. Zwefel, A. Shaffer, J. J. Tal-
ley, J. L. Masferrer, K. Serbert, P. C. Isakson, Proc. Natl. Acad. Sci. USA 95 (1998) 13313
3. T. D. Warner, F. Giuliano, I. Vaynovie, A. Bukasa, J. A. Mitchell, J. R. Vave, Proc. Natl.
Acad. Sci. USA 96 (1999) 7563
4. L. J. Marnett, A. S. Kalgutkar, Trends Pharmacol. Sci. 20 (1999) 465
5. G. Dannhardt, W. Kiefer, Eur. J. Med. Chem. 36 (2001) 109
6. L. J. Marnett, A. S. Kalgutkar, Curr. Opin. Chem. Biol. 2 (1998) 482
7. P. Parsit, D. Reindeau, Annu. Rep. Med. Chem. 32 (1997) 211
8. A. G. Habeeb, P. N. P. Rao, E. D. Knaus, J. Med. Chem. 44 (2001) 2921
9. C. A. Imansa, J. Alfon, A. F. de Arriba, F. L. Cavalcanti, I. Escamilla, L. A. Gomez, A.
Mirralles, R. Solvia, J. Bartroli, E. Carceller, M. Merlos, J. G. Rafanell, J. Med. Chem. 46
(2003) 3643
10. L. M. Jackson, C. J. Hawkey, Exp. Opin. Invest. Drugs 8 (1999) 963
11. C. Hawkey, L. Laine, T. Simon, A. Beaulieu, J. Maldonado-Cocco, E. Acevedo, A. Sha-
hane, H. Quan, J. Bolognese, E. Mortensen, Arthritis Rheum. 43 (2000) 370
12. V. K. Tammara, M. M. Narurkar, A. M. Crider, M. A. Khan, J. Pharm. Sci. 83 (1994) 644
13. J. M. Dogne, C. T. Supuran, D. Pratico, J. Med. Chem.48 (2005) 2251
14. D. Galanakis, A. P. Kourounakis, K. C. Tsiakitzis, C. Doulgkeris, E. A. Rekka, A. Gava-
las, C. Kravaritou, C. Charitos, P. N. Kourounakis, Bioorg. Med. Chem. Lett. 14 (2004) 3639
15. A. S. Kalgutkar, A. B. Marnett, B. C. Crews, R. P. Remmel, J. Med. Chem. 43 (2000)
2860
16. M. Duflos, M. R. Nourrisson, J. Brelet, J. Courant, G. Le Baut, N. Grimaud, J. Y. Petit,
Eur. J. Med. Chem. 36 (2001) 545
17. A. S. Kalgutkar, B. Crews, S. Rowlinson, C. Garner, K. Seibert, L. Marnett, Science 280
(1998) 1268
18. M. Tuncbilek, N. Altanlar, Farmaco 54 (1999) 475
19. K. Mogilaiah, R. B. Rao, K. N. Reddy, Ind. J. Chem. B 38 (1999) 818
20. M. S. Y. Khan, G. Chawla, M. A. Mueed, Ind. J. Chem. B 43 (2004) 1302
21. C. A. Winter, E. A. Risley, G. W. Nuss, Proc. Soc. Exp. Biol. Med. 111 (1962) 544
22. O. O. Adeyemi, O. S. Okpo, O. J. Okpaka, J. Ethnopharmacol. 90 (2004) 45
23. V. Cioli, S. Putzolu, V. Rossi, S. P. Barcellona, C. Corradino, Toxicol. Appl. Pharmacol.
50 (1979) 283
24. H. Ohkawa, M. Ohishi, K. Yagi, Anal. Biochem. 95 (1979) 351
25. Y. Naito, T. Yoshikawa, N. Yoshinda, M. Kondo, Dig. Dis. Sci. 43 (1998) 30s.
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