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ae

Medicinal Chemistry, 2018, 14, 674-687

RESEARCH ARTICLE
ISSN: 1573-4064
eISSN: 1875-6638

Impact
Factor:

Diclofenac 1,3,4-Oxadiazole Derivatives; Biology-Oriented Drug Synthesis

2.631

(BIODS) in Search of Better Non-Steroidal, Non-Acid Antiinflammatory

Agents

Shazia Shaha,c, Arshiaa, Nida Siraj Kazmia, Almas Jabeena, Aisha Faheemb, Nida Dastagirb, Tariq
Ahmedb, Khalid Mohammed Khana,*, Shakil Ahmedd, Abeer Razaa and Shahnaz Perveene

a
H. E. J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Ka-
rachi, Karachi-75270, Pakistan; bDr. Panjwani Center for Molecular Medicine and Drug Research, International Cen-
ter for Chemical and Biological Sciences, University of Karachi, Karachi-75270, Pakistan; cDepartment of Chemistry,
Federal Urdu University of Art, Science and Technology, Pakistan; dIndustrial Analytical Center at H. E. J. Research
Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi-
75270, Pakistan; ePCSIR Laboratories Complex, Karachi, Shahrah-e-Dr. Salimuzzaman Siddiqui, Karachi-75280,
Pakistan
Abstract: Background: Inflammation is defined as the response of immune system cells to dam-
aged or injured tissues. The major symptoms of inflammation include increased blood flow, cellu-
lar influx, edema, elevated cellular metabolism, reactive oxygen species (ROS) nitric oxide (NO)
and vasodilation. This normally protective mechanism against harmful agents when this normal
mechanism becomes dysregulated that can cause serious illnesses including ulcerative colitis,
Crohn’s disease, rheumatoid arthritis, osteoarthritis, sepsis, and chronic pulmonary inflammation.
Method: In this study synthetic transformations on diclofenac were carried out in search of better
non-steroidal antiinflammatory drugs (NSAIDs), non-acidic, antiinflammatory agents. For this
purpose diclofenac derivatives (2-20) were synthesized from diclofenac (1). All derivatives (2-20)
and parent diclofenac (1) were evaluated for their antiinflammatory effect using different parame-
ARTICLE HISTORY ters including suppression of intracellular reactive oxygen species (ROS), produced by whole
blood phagocytes, produced by neutrophils, and inhibition of nitric oxide (NO) production from
Received: July 31, 2017
Revised: February 14, 2018 J774.2 macrophages. The most active compound also evaluated for cytotoxicity activity.
Accepted: March 08, 2018
Results: Diclofenac (1) inhibited the ROS with an IC 50 of 3.9 ± 2.8, 1.2 ± 0.0 μg/mL respectively
DOI:
10.2174/1573406414666180321141555
and inhibited NO with an IC50 of 30.01 ± 0.01 μg/mL. Among its derivatives 4, 5, 11, 16, and 20,
Medicinal Chemistry

showed better antiinflammatory potential. The compound 5 was found to be the most potent in-
hibitor of intracellular ROS as well as NO with IC50 values of 1.9 ± 0.9, 1.7 ± 0.4 μg/mL respecti-
vely and 7.13 ± 1.0 μg/mL, respectively, and showed good inhibitory activity than parent di-
clofenac. The most active compounds were tested for their toxic effect on NIH-3T3 cells where all
compounds were found to be non-toxic compared to the standard cytotoxic drug cyclohexamide.
Conclusion: Five derivatives were found to be active. Compound 5 was found to be the most po-
tent inhibitor of ROS and NO compared to parent diclofenac 1 and standard drugs ibuprofen and
L-NMMA, respectively. The most active compounds 1, 4, 5, 11 and 20 were found to be non-toxic
on NIH-3T3 cells. Compound 4, 5, and 20 also showed good antiinflammatory potential, com-
pound 11 and 16 showed moderate and low level of inhibition, respectively.
Keywords: Synthesis, diclofenac, 1,3,4-oxadiazole, antiinflammatory parameters, ROS suppression, and NO inhibition.

1. INTRODUCTION tion of leucocytes such as neutrophils and macrophages from

Inflammation is defined as the response of immune sys- blood to damaged cells, irritants and pathogens etc for their
tem cells to damaged or injured tissues that involve migra- repair and removal [1]. During inflammation, inflammatory
mediators including reactive oxygen and nitrogen species,
leukotrienes, prostaglandins, cytokines etc are released that
increases the vascular permeability and leucocytes migration
*Address correspondence to this author at the H. E. J. Research Institute of
Chemistry, International Center for Chemical and Biological Sciences, to the site of inflammation [2]. The major symptoms of in-
University of Karachi, Karachi-75270, Pakistan; Tel: 0092-21-34824910; flammation include increased blood flow, cellular influx,
Fax: 0092-21-34819018-9; E-mails: [email protected];
[email protected] edema, elevated cellular metabolism reactive oxygen species

1875-6638/18 $58.00+.00 © 2018 Bentham Science Publishers

Diclofenac 1,3,4-Oxadiazole Derivatives Medicinal Chemistry, 2018, Vol. 14, No. 7 675

Cl O OH
O
Cl
NH O
N
Cl OH O
O OH

Diclofenac O Aspirin

Indometacin

O OH
F
F H
N OH
F
O

Flufenamic acid Ibuprofen

Fig. (1). Reported antiinflammatory drug having acidic moiety.

(ROS) nitric oxide (NO) and vasodilation [3]. This normally marketed antiinflammatory drug (diclofenac) were carried
protective mechanism against harmful agents when this out and evaluate their antiinflammatory potentials. As 1,3,4-
normal mechanism becomes dysregulated that can cause oxadiazole also possess good antiinflammatory activity [23,
serious illnesses including ulcerative colitis, Crohn’s disease, 24]. To the best of our knowledge only compounds 1-4 were
rheumatoid arthritis, osteoarthritis, sepsis, and chronic pul- previously reported while remaining analogs are new.
monary inflammation [4].
2. MATERIALS AND METHOD
The most favored class of drug in use for controlling
various pathological conditions is the non-steroidal anti- Methanol, hydrazine hydrate, carbon disulfide, phenacyl
inflammatory drugs (NSAIDs) [5]. The mode of action of halide, aryl halides, and potassium hydroxide were pur-
NSAIDs involves the repression of prostaglandin biosynthe- chased from Sigma-Aldrich, USA and used as received
sis from arachidonic acid by inhibiting the enzyme cy- without purification. Diclofenac was donated by a local
clooxygenases (COXs) [6]. Among the most admired pharmaceutical company. TLC plates were pre-coated silica
NSAIDs is diclofenac sodium. This drug is accepted in more gel aluminium plates (Kieselgel 60, 254, E. Merck). Visuali-
than 120 countries across the world, and is ranked 30th zation of TLC chromatograms was carried out under UV
among the top 200 drugs [7]. light at wavelengths of 254 and 365 nm. Melting points of all
Chemical name of diclofenac is 2-(2, 6-dichloranilino) the compounds were determined on Stuart® SMP10 melting
phenylacetic acid. Diclofenac has potential biological activi- point apparatus and are uncorrected. EI-MS has recorded on
ties including antibacterial [8], antitumour [9], anticancer a Finnigan MAT-311A (Germany) mass spectrometer. 1H-
[10, 11], however, the major use of diclofenac is as an NMR experiments were carried out on Avance Bruker AM
antiinflammatory drug [11, 12]. The suppressive effect of 300, 400, and 500 MHz machines. 13C-NMR experiments
diclofenac on ROS produced from rat peritoneal neutrophils were carried on Avance Bruker AM 300, 400 and 500 MHz
was also reported previously [13]. Unfortunately, all instruments at 75, 100, and 125 MHz, respectively. Infrared
NSAIDs drugs such as diclofenac, indomethacin, aspirin, (IR) spectra were recorded on JASCO IR-A-302 spectropho-
flufenamic acid, phenylbutazone, and ibuprofen have gastro- tometer.
intestinal tract side effects which is mainly due to the contact
of open carboxylic moiety (pKa = 3.5-5.5) in these drugs to 3. EXPERIMENTAL
gastrointestinal tract [14, 15] (Fig. 1). Therefore, our re-
search group continuously engaged to combat this evil dis- 3.1. Antiinflammatory Assays
ease in an efficient manner by synthesizing different func- 3.1.1. Oxidative Burst Inhibition by Chemiluminescence
tionally oriented molecules and evaluate their antiinflam- Technique
matory activities in past [16-21].
Oxidative burst studies using luminol enhanced chemi-
BIODS science consists of some modification of previ- luminescence technique were performed as described by
ously known drug to reduce its side effect, increase activity Mesaik et al. with some modifications [25]. The assay was
potential etc. In the current study, we used the BIODS ap- performed on whole blood and neutrophils isolated from
proach [22], syntheses of 1,3,4-oxadiazole derivatives of blood of healthy human volunteers using luminol as a probe
676 Medicinal Chemistry, 2018, Vol. 14, No. 7 Shah et al.

and zymosan as an activator. Briefly 25 L of diluted whole 3.1.4.2. Diclofenac Methyl Ester (2)
blood in HBSS++ or 25 L of isolated neutrophils (1 x 106
Diclofenac acid (5 g) was dissolved in methanol (80 mL)
cells/mL) were incubated with 25 L of different concentra-
with stirring and sulfuric acid (3 mL) was added drop wise
tions of compounds (1, 10, 25 and 100 g/mL) each in tripli-
with continuous stirring and the mixture was heated under
cate in white half area 96 well plates (Costar, NY, USA).
The plates were incubated at 37 °C for 15 min in the thermo- reflux for 15 min. After completion of the reaction (TLC
analysis), it was cooled to room temperature and poured onto
stat chamber of a luminometer (Labsystems, Helsinki, Fin-
crushed ice (250 g) to obtain crystalline solid (2) which was
land). 25 L of (7 x 10-5 M) luminol (Research Organics,
filtered, washed with water followed by 5% sodium hydro-
Cleveland, OH, USA) and, 25 L serum opsonized zymosan
gen carbonate solution, finally with water and dried in air.
(SOZ) 2 mg/mL (Fluka, Buchs, Switzerland) was then added
into the wells. The plates were then read in luminometer for 3.1.4.3. Diclofenac Hydrazide (3)
50 min and results were recorded as total integral readings as
relative light units (RLU) [25]. Diclofenac ester (2 g) was dissolved in 20 mL methanol then
hydrazine hydrate (10 mL) was added while keeping in ice
3.1.2. Nitric Oxide Assay bath and stirred for 48 h. Progress of the reaction was moni-
The mouse macrophage cell line J774.2 (European Col- tored through TLC. A solid was formed when poured on to
lection of Cell Cultures, UK) was cultured in 75 mL flasks ice, filtered and dried in air.
IWAKI (Asahi Techno Glass, Japan) in DMEM Sigma- 3.1.4.4. Diclofenac Oxadiazole-2-thione (4)
Aldrich (Steinheim, Germany) that contained 10% fetal bo-
vine serum GIBCO (NY US) supplemented with streptomy- To a solution of diclofenac hydrazide (1.0 g in 50 mL of
cin/penicillin 1%. Flasks were kept at 37 °C in atmosphere ethanol), potassium hydroxide (0.5 g) and carbon disulfide
of humidified air containing 5% CO2, cells were seeded in (12 mL) were added drop wise at room temperature and the
96-well plate (1 x 106 cells/mL), and were stimulated by 30 reaction was allowed to reflux for 24 h. After completion of
g/mL E. coli lipopolysaccharide (LPS) (DIFCO Laborato- reaction (TLC analysis), a potassium salt of the product was
ries Michigon, USA) and test compounds were added at obtained which was acidified with HCL (conc.) with stirring
three different concentrations (1, 10, 100 g/mL). The plates condition to obtain a solid product which was filtered,
were incubated at 37 °C in 5% CO2 and humidified air for 48 washed with distilled water, and dry at room temperature.
hours. The cell culture supernatants were collected. nitrite 3.1.4.5. Derivatives of Diclofenac Oxadiazole Thione (5-20)
accumulation in J774.2 cell culture supernatant was meas-
ured using the Griess method described previously [26]. Diclofenac oxadiazole thione (0.352 g), ethanol (15 mL),
triethyl amine (0.1 mL) and phenacyl halide and other hal-
3.1.3. MTT Cytotoxicity Assay
ides (1 mmol) were added in a round-bottomed flask and
Cytotoxicity of compounds on NIH-3T3 fibroblast cells refluxed at 100 °C for 5 h. After completion of the reaction,
(ATCC, Manassas, USA) was evaluated by using the stan- mixture was placed in refrigerator to obtain precipitates. The
dard MTT colorimetric assay. Briefly, 100 L of 6  104 solid was filtered, washed with water and dried in vacuum.
cells/mL in DMEM supplemented with 10% FBS were
plated into 96-wells flat bottom plate and incubated over- 4. RESULT AND DISCUSSION
night at 37°C in 5% CO2. Three different concentrations of
test compound (1, 10 and 100 g/mL) were added to the 4.1. Chemistry
plate in triplicates and incubated for 48 h. 50 L of 0.5 Diclofenac derivative was synthesized by different steps
mg/mL MTT was added to each well and plates were then under refluxing conditions. The diclofenac (1) in the pres-
further incubated for 4 h. MTT was aspirated and 100 L of ence of methanol and sulphuric acid was converted into
DMSO was then added to each well. The extent of MTT methyl ester 2 and then this ester converted hydrazide 3 with
reduction to formazan within cells was calculated by measur- hydrazine hydrate in methanol. Resulting hydrazide 3 was
ing the absorbance at 540 nm, using spectrophotometer transformed to oxadiazole derivative 4 in carbon disulfide
(Spectra Max plus, Molecular Devices, CA, USA). The cyto- under basic conditions. Final products 5-20 were obtained by
toxic activity was recorded as concentration causing 50% reaction of oxadiazole derivative with different phenacyl
growth inhibition (IC50) for 3T3 cells [27]. halides and aryl halides in the presence of ethanol and
triethyl amine (Scheme 1). Chemical structures of all of
3.1.4. General Procedure these derivatives were established by 1H-NMR, 13C-NMR,
EI-MS, and IR spectroscopy.
3.1.4.1. Diclofenac (1)
Diclofenac sodium (5 g) was dissolved in water with stir- 4.2. Antiinflammatory Activity of Diclofenac (1) and its
Derivatives
ring followed by the addition of 2 mL of 2 N HCl drop wise
to obtain the precipitate of diclofenac acid then addition of Diclofenac 1 and all derivatives (Fig. 2) were initially
ice water for the termination of reaction. A solid was formed, tested to check their inhibitory potential on intracellular reac-
filtered, washed with cold water and dry at room tempera- tive oxygen species (ROS) produced from human whole
ture. The structure was identified by using spectroscopic blood phagocytes at single concentration of 25 g/mL. The
techniques including, 1H-, 13C-NMR, EI-MS, ESI-MS, compounds showing inhibitory potential were then tested on
HRMS, and IR. two different parameters of inflammation. The active com-
Diclofenac 1,3,4-Oxadiazole Derivatives Medicinal Chemistry, 2018, Vol. 14, No. 7 677

5 Cl
6 5
4 6 Cl 5 Cl
1
CH3OH 4
NH2-NH2.H2O 4
6
CS2, KOH
3 1
2 NH Conc. H2SO4 3
3
1
2 NH NH
Cl 12 13 2
14 OH Cl 12 13 14
7 11 O Me Cl 12 13
14 NHNH2
7 11 7 11
8 10 O
9
8 10 O 8 10 O
9
(1) 9
(2) (3)
5 Cl 5
4
6 6 Cl
4
1 1
3
NH R-X 3
2 N N 2 NH
12 13 15 N N R
Cl Cl 12 13
7 SH S
14 O 7 14 O 15
8 10 8 10
9 9
(4) (5-20)

Scheme 1. Syntheses of oxadiazole derivatives of dicleofenac.

Fig. (2). Structure-activity relationship of diclofenac and oxadiazole 4 and compound 5.

pounds were further evaluated for their inhibitory effect on into ester 2 and hydrazide 3 the activity was found to be di-
the production of ROS from isolated neutrophils and on ni- minished. Conversion of hydrazide 3 into its oxadiazole de-
tric oxide (NO) generated from lipopolysaccharide (LPS) rivative 4 it regains the inhibitory activity on ROS f 3.1.4.1.
stimulated J774.2 macrophages (Table 1). rom whole blood and neutrophils with IC50 values of 5.7 ±
1.5 and 6.7 ± 0.7 g/mL, respectively. However potent inhi-
In this study diclofenac 1 inhibited the ROS with an IC50 bition of NO IC50 9.34 ± 0.8 g/mL was observed as compa-
value 3.9 ± 2.8 and 1.2 ± 0.0 g/mL from human whole blo- red to parent 1 as well as standard L-NMMA. Conversion of
od cells and isolated neutrophils, respectively. It also mode- 4 into 5 gave a most potent inhibitior of ROS as well as NO
rately inhibited NO with an IC50 value of 30.1 ± 0.0 g/mL of this series having an IC50 of value 1.9 ± 0.9 and 1.7 ± 0.4
compared to the standard L-NMMA IC50 24.2 ± 0.8 g/mL. g/mL on whole blood and neutrophils, respectively, and
However, when the acid group of diclofenac was converted 7.13 ± 1.0 g/mL on nitric oxide (NO) (Fig. 3).
678 Medicinal Chemistry, 2018, Vol. 14, No. 7 Shah et al.

Fig. (3). Structure-activity relationship of phenacyl analogs of substituted oxadiazole dicleofenac 5, 11, and 16.

The oxadiazole derivative of diclofenac 4 reacts with dif- found to be non-toxic as compared to the cyclohexamide
ferents phenacyl halide to afford compounds 5-17. Phenacyl which was used as a standard cytotoxic drug (Table 2).
derivative of 1,3,4-oxadiazole diclofenac 5 was found to be
Chloro, nitro, methoxy and phenyl substituted phenacyl
most active compound which showed superior antiinflam-
oxadiazoles 7, 8, 9, 12-15, and 17, respectively were found
matory activity than parent diclofenac 1 as well as standard
to be inactive. 1,3,4-Oxadiazole diclofenac 4 was also reac-
such as ibuprofen which is used for ROS inhibition having
ted with varyingly substituted aryl halide and afforded
an IC50 values of 11.2 ± 1.9 and 2.5 ± 0.6 g/mL for whole
dcompounds 18, 19, and 20. Among them only compound 20
blood and isolated neutrophils, respectively, and from L- having a 2-bromo benzyl substituent showed good activity
NMMA which was used as standard for nitric oxide (NO)
with IC50 values of 4.5 ± 1.7, 3.4 ± 0.5, and 21.9 ± 1.15
inhibition IC50 = 24.2 ± 0.8 g/mL. Compound 11, a substi-
g/mL for whole blood, neutrophil ROS and NO inhibition,
tuted phenacyl derivative of oxadiazole having a bromo resi-
respectively. While chloro substituted derivatives 18 and 19
due at position 3 showed moderate level of inhibition having
were found to be inactive (Figs. 5, 6, 7).
an IC50 value of 21.7 ± 11.0, 5.0 ± 0.9 and 14.8 ± 2.0 g/mL
for whole blood, neutrophils ROS, and nitric oxide inhibiti-
on, respectively. While compound 10 having a bromo group 5. SPECTROSCOPIC DATA OF SYNTHETIC COM-
at postion 4 was found to be inactive. Hydroxy substituted POUNDS
phenacyl derivative 16 showed low level of inhibition on
5.1. 2-(2-(2,6-Dichlorophenylamino)phenyl)acetic Acid
whole blood ROS and neutrophil ROS IC50 = 31.2 ± 5.2, 9.0
(1)
± 1.0 g/mL respectively. Where as moderate inhibition of
nitric oxide IC50 = 26.2 ± 3.5 g/mL was observed (Fig. 4). M.p. 152-154°C (lit. 153-154°C [28]); Rf: 0.31 (ethyl a-
The cytotoxicites of most active derivatives 1, 4, 5, 11 and cetate/hexanes, 2:8); 1H-NMR (400 MHz, DMSO-d6):  12.7
20 were tested on NIH-3T3 cells where all compounds were (s, 1H, OH), 7.52 (d, 2H, J(3,4),(5,4) = 7.8 Hz, H-3, H-5),
Diclofenac 1,3,4-Oxadiazole Derivatives Medicinal Chemistry, 2018, Vol. 14, No. 7 679

Fig. (4). Structure-activity relationship of benzyl analogs of oxadiazole dicleofenac 20.

Fig. (5). 1,3,4-Oxadiazole derivative of dicleofenac.

Table 1. Anti-inflammatory activity of diclofenac (1) and its derivatives (2-20).

ROS Inhibition ROS Inhibition NO Inhibition

Compound Structure Whole Blood Neutrophils IC50 ± SD
IC50 ± SD (g/mL) IC50 ± SD (g/mL) (g/mL)

1 3.9 ± 2.8 1.2 ± 0.0 30.0 ± 0.0

2 NAb NDc NDc

Table 1. contd…
680 Medicinal Chemistry, 2018, Vol. 14, No. 7 Shah et al.

ROS Inhibition ROS Inhibition NO Inhibition

Compound Structure Whole Blood Neutrophils IC50 ± SD
IC50 ± SD (g/mL) IC50 ± SD (g/mL) (g/mL)

3 NAb NDc NDc

4 5.7 ± 1.5 6.7 ± 0.7 9.34 ± 0.8

5 1.9 ± 0.9 1.7 ± 0.4 7.13 ± 1.0

6 NAb NDc NDc

7 NAb NDc NDc

8 NAb NDc NDc

9 NAb NDc NDc

10 NAb NDc NDc

11 21.7 ± 11.0 4.9 ± 0.9 14.8 ± 2.0

12 NAb NDc NDc

Table 1. contd…
Diclofenac 1,3,4-Oxadiazole Derivatives Medicinal Chemistry, 2018, Vol. 14, No. 7 681

ROS Inhibition ROS Inhibition NO Inhibition

Compound Structure Whole Blood Neutrophils IC50 ± SD
IC50 ± SD (g/mL) IC50 ± SD (g/mL) (g/mL)

13 NAb NDc NDc

14 NAb NDc NDc

15 NAb NDc NDc

16 31.2 ± 5.2 9.0 ± 1.0 26.2 ± 3.5

17 NAb NDc NDc

18 NAb NDc NDc

19 NAb NDc NDc

20 4.5 ± 1.7 3.4 ± 0.5 21.9 ± 1.1

SDa means Standard deviation; NAb = Not Active; NDc = Not determined; Standard: Ibuprofen, (whole blood) IC50 = 11.2 ± 1.9 g/mL; (neutrophil) IC50 = 2.5
± 0.6 g/mL; N G monomethyl L-arginine acetate (L-NMMA) nitric oxide IC50= 24.2 ± 0.8.

Table 2. Cytotoxicity of active compounds on NIH-3T3 cells. The IC50 values were calculated using three doses (1, 10, 100 g/mL)
of each compound. Values are expressed as mean ± SD of experiments done in triplicate. Results were compared with
cyclohexamide as a standard cytotoxic drug.

Compounds IC50 ± SD a (g/mL)

1 29.8 ± 0.4

4 32.0 ± 0.6
Table 2. contd…
682 Medicinal Chemistry, 2018, Vol. 14, No. 7 Shah et al.

Compounds IC50 ± SD a (g/mL)

5 76.7 ± 6.0

11 >100

20 >100

Cyclohexamide 0.13 ± 0.02

SDa means Standard deviation.

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Fig. (6). The graph represents the inhibitory effect of compounds on oxidative burst. Compounds were tested at three different concentrations
(1, 10 and 100 g/mL). Results are presented in relative light units (RLU) and oxidative burst activity of whole blood phagocytes using lumi-
nol as a probe. Each vertical bar represents a mean of triplicate. Results were compared to the control. Where Control is zymosan activated
whole blood phagocytes with no drug.

70 50 mg/mL 5 mg/mL 0.5 mg/mL

60

50
mM)
Nitric oxide (m

40

30

20

10

0
1 4 5 11 16 20 Control Blank
Compounds

Fig. (7). The graph represents the inhibitory effect of compounds on Nitric oxide (NO) production from mouse macrophage J774.2 cell line.
Compounds were tested at three different concentrations (0.5, 5 and 50 g/mL). Each vertical bar represents a mean of triplicate. Results were
compared to the control. Where Control = Cells activated by lipopolysaccharide (LPS) with no drug and Blank = Cells without LPS.

7.23 (s, 1H, NH), 7.20 (m, 2H, H-10, H-11), 7.07 (t, 1H, (C-O); Anal. Calcd for C14H11Cl2NO2: C, 56.78; H, 3.74; Cl,
J(4/3,5) = 8.0 Hz, H-4), 6.86 (t, 1H, J(9/10,11) = 8.0 Hz, H-9), 23.94; N, 4.73; O, 10.80; Found: C, 56.77; H, 3.73; N, 4.72.
6.28 (d, 1H, J (8,9) = 8.0 Hz, H-8), 3.68 (s, 2H, CH2); EI-MS:
m/z (rel. abund. %), 298 [M+2]+ (29.4.7), 296 [M] + (18.5), 5.2. Methyl 2-(2-(2,6-dichlorophenylamino)phenyl)ace-
278 (4.7), 242 (40.0), 214 (100.0), 179 (26.3), 151 (12.5), tate (2)
109 (6.9); HREI-MS: m/z Calcd for C14H11Cl2NO2 [M] +
Yield: 96 %; m.p. 98-100°C (lit. 97-99°C [28]); Rf: 0.36
296.1486, Found 296.1485; IR (KBr, cm-1): 3347 (O-H),
(ethyl acetate/hexanes, 2:8); 1H-NMR (400 MHz, DMSO-
3362 (N–H), 1670 (C=O), 1569 (C=C), 1259 (C-N), 1180
Diclofenac 1,3,4-Oxadiazole Derivatives Medicinal Chemistry, 2018, Vol. 14, No. 7 683

d6):
7.53 (d, 2H, J(3,4),(5,4) = 8.0 Hz, H-3, H-5), 7.22 (m, 2H, NH), 7.06 (t, 1H, J(4/3,5) = 7.5 Hz, H-4) 6.83 (t, 1H, J(9/8,10) =
H-9, H-10), 7.06 (d, 1H, J (11,10) = 6.0 Hz, H-11), 7.03 (t, 1H, 7.5 Hz, H-9), 6.19 (d, 1H, J(8,9) = 8.0 Hz, H-8), 5.05 (s, 2H,
J(4/3,5) = 7.6 Hz, H-4), 6.25 (d, 1H, J (8,9) = 8.0 Hz, H-8), 3.77 16-CH2), 4.35 (s, 2H, 13-CH2); ESI-MS m/z: 470.0 (M+);
(s, 2H, CH2), 3.64 (s, 3H, OCH3); EI-MS: m/z (rel. abund. HR-ESI-MS m/z Calcd for C23H17Cl2N3O2S [M]+ 470.3710,
%), 311 [M+2]+ (24.7), 309 [M] + (36.7), 277 (9.6), 242 found 470.3711; IR (KBr, cm-1): 3214 (N-H), 1586 (C=N),
(39.2), 214 (100.0), 179 (10.2), 151 (7.6); HREI-MS: m/z 1509 (C=C), 1489 (C=S), 1170 (C-O); Anal. m/z Calcd
calcd for C15H13Cl2NO2 [M]+ 309.0323, Found 309.0322; IR C23H17Cl2N3O2S: C, 58.73; H, 3.64; Cl, 15.07; N, 8.93; O,
(KBr, cm-1): 3382 (N-H), 1660 (C=O), 1579 (C=N), 1569 6.80; S, 6.82 Found: C, 58.72; H, 3.63; N, 8.92.
(C=C), 1259 (C-N), 1170 (C-O); Anal. Calcd for
C15H13Cl2NO2: C, 58.08; H, 4.22; Cl, 22.86; N, 4.52; O, 5.6. 3-(3,4-Dichlorobenzyl)-5-(2-(2,6-dichlorophenylami-
10.32; Found: C, 58.07; H, 4.21; N, 4.51. no)benzyl)-1,3,4-oxadiazole-2(3H)-thione (6)
Yield: 91%; m.p 120-122°C; Rf: 0.51 (ethyl aceta-
5.3. 2-(2-(2,6-Dichlorophenylamino)phenyl)acetohydra-
te/hexane, 2:8) 1H-NMR (400 MHz, DMSO-d6):
7.68 (d,
zide (3)
1H, J(18, 21) = 1.2 Hz, H-18), 7.52 (s, 1H, NH), 7.50 (d, 2H,
Yield: 97%; m.p. 132-134°C C (lit. 134-136°C [29]); Rf: J(3,4),(5,4) = 8.0 Hz, H-3, H-5), 7.36 (dd, 1H, J(22,18) = 1.2 Hz
0.35 (ethyl acetate/hexanes, 2:8); 1H-NMR (400 MHz, DM- J(22,21) = 8.4 Hz, H-22) 7.23 (t, 1H, J(4/3,5) = 8.0 Hz, H-4), 7.16
SO-d6):
9.46 (s, 1H, NH), 9.51 (s, 1H, NH), 7.51 (d, 2H, (d, 1H, J(11, 10) = 7.6 Hz, H-11), 7.09 (m, 2H, H-8, H-10),
J(3,4),(5,4) = 8.4 Hz, H-3, H-5), 7.16 (m, 2H, H-10, H-11), 7.04 6.85 (t, 1H, J(9/8,10) = 7.6 Hz, H-9), 6.19 (d, 1H, J(21,22) = 8.1
(t, 1H, J(4/3,5) = 7.6 Hz, H-4), 6.85 (t, 1H, J(9/10,11) = 7.6 Hz, H- Hz, H-21), 4.44 (s, 2H, 16-CH2), 4.35 (s, 2H, 13-CH2); 13C-
9), 6.29 (d, 1H, J (8,9) = 8.0 Hz, H-8), 4.30 (s, 2H, CH2), 3.50 NMR: (100 MHz, DMSO-d6):
166.5 (C=S), 162.4 (C-14),
(s, 2H, NH2); 13C-NMR: (100 MHz, DMSO-d6):
C, 170.7 142.9 (C-2, C-6), 138.1 (C-17), 136.8 (C-19), 131.5 (C-20),
(C=O), 142.9 (C-1), 137.1 (C-7), 130.2 (C-3, C-5),), 129.2 130.9 (C-18), 130.8 (C-21), 130.6 (C-22), 130.5 (C-3, C-5),
(C-3, C-5), 127.1 (C-8), 125.3 (C-9), 124.8 (C-10), 120.6 (C- 130.2 (C-7), 129.2 (C-1), 129.1 (C-11), 128.2 (C-9), 126.4
10), 116.0 (C-4), 37.6 (CH2); EI-MS: m/z (rel. abund. %), (C-10), 122.2 (C-8), 120.4 (C-12), 115.5 (C-4), 34.2 (13-
311 [M+2] + (2.4), 309 [M]+ (3.8), 278 (58.2), 242 (17.2), CH2), 27.3 (16-CH2); ESI-MS m/z: 511.0 (M+); HR-ESI-MS
214 (100.0), 179 (13.0), 151 (6.9); HREI-MS: m/z Calcd for m/z Calcd for C22H15Cl4N3OS [M]+ 515.2510, Found
C14H13Cl2N3O [M]+ 309.0436, Found 309.0435; IR (KBr, 511.2511; IR (KBr, cm-1): 3211 (N-H), 1545 (C=N), 1543
cm-1): 3387 (N-H), 1670 (C=O), 1599 (C=N), 1560 (C=C), (C=C), 1430 (C=S), 1187 (C-O); Anal. m/z Calcd
1299 (C-N); Anal. Calcd for C14H13Cl2N3O: C, 54.21; H, C22H15Cl4N3OS: C, 51.68; H, 2.96; Cl, 27.74; N, 8.22; O,
4.22; Cl, 22.86; N, 13.55; O, 5.16; Found: C, 54.20; H, 4.21; 3.13; S, 6.27, Found: C, 51.67; H, 2.95; N, 8.21.
N, 13.54.
5.7. 1-(4-Chlorophenyl)-2-(5-(2-(2,6-dichlorophenylami-
5.4. 5-(2-(2,6-Dichlorophenylamino)benzyl)-1,3,4-oxadia- no)benzyl)-2-thioxo-1,3,4-oxadiazol-3(2H)-yl)ethanone
zole-2-thiol (4) (7)
Yield: 97%; m.p. 160-162°C (lit. 162-164°C [30]); Rf: Yield: 91%; m.p 118-120°C; Rf: 0.50 (ethyl aceta-
0.38 (ethyl acetate/hexanes, 2:8); 1H-NMR (400 MHz, DM- te/hexanes, 2:8); 1H-NMR (300 MHz, DMSO-d6):
8.03 (d,
SO-d6):
14.28 (s, 1H, SH), 7.52 (d, 2H, J(3,4),(5,4) = 8.4 Hz, 2H, J(20,19),(22,23) = 8.4 Hz, H-20, H-22), 7.64 (d, 2H,
H-3, H-5), 7.23 (m, 2H, H-10, H-11), 7.12 (s, 1H, NH), 7.09 J(19,20),(23,22) = 8.4 Hz, H-19, H-23), 7.53 (d, 2H, J(3,4),(5,4) = 8.1
(t, 1H, J(4/3,5) = 6.9 Hz, H-4), 6.86 (t, 1H, J(9/10,11) = 7.2 Hz, H- Hz, H-3, H-5), 7.24 (t, 1H, J(10/9,11) = 7.5 Hz, H-10), 7.16 (d,
9), 6.19 (d, 1H, J (8,9) = 8.0 Hz, H-8), 4.24 (s, 2H, CH2); 13C- 1H, J (11,10) = 7.5 Hz, H-11), 7.12 (s, 1H, NH), 7.08 (t, 1H,
NMR: (100 MHz, DMSO-d6):
177.8 (C-14), 162.5 (C-13), J(4/3,5) = 6.9 Hz, H-4), 6.84 (t, 1H, J(9/8,10) = 6.0 Hz, H-9), 6.21
143.1 (C-7), 136.9 (C-1), 131.8 (C-1), 131.0 (C-2, C-6), (d, 1H, J (8,9) = 8.1 Hz, H-8), 5.03 (s, 2H, 16-CH2), 4.35 (s,
129.1 (C-3, C-5), 128.3 (C-4), 126.4 (C-8), 121.3 (C-12), 2H, 13-CH2); ESI-MS m/z: 504.0 (M+); HR-ESI-MS m/z
120.3 (C-9), 115. (C-10), 27.6 (CH2); EI-MS: m/z (rel. a- Calcd for C23H16Cl3N3O2S [M]+ 504.8160, Found 504.8161;
bund. %), 353 [M+2]+ (24.3), 351 [M] + (32.6), 291 (63.6), IR (KBr, cm-1): 3221 (N–H), 1678 (C=O), 1562 (C=N), 1528
278 (54.6), 256 (19.6), 214 (100.0), 131 (35.4); HREI-MS: (C=C),1423 (C=S), 1026 (C–O); Anal. m/z Calcd for for for
m/z Calcd for C15H11Cl2N3OS [M]+ 352.2383, Found C23H16Cl3N3O2S: C, 54.72; H, 3.19; Cl, 21.07; N, 8.32; O,
352.2382; IR (KBr, cm-1): 3367 (N-H), 2979 (S-H), 1579 6.34; S, 6.35 Found: C, 54.71; H, 3.18; N, 8.33.
(C=N), 1500 (C=C), 1294 (C-N), 1286 (C=S); Anal. Calcd
for C15H11Cl2N3OS: C, 51.15; H, 3.15; Cl, 20.13; N, 11.93; 5.8. 1-(3,4-Dichlorophenyl)-2-(5-(2-(2,6-dichlorophenyl-
O, 4.54; S, 9.10; Found: C, 51.14; H, 3.14; N, 11.92. amino)benzyl)-2-thioxo-1,3,4-oxadiazol-3(2H)-yl)ethano-
ne (8)
5.5. 2-(5-(2-(2,6-Dichlorophenylamino)benzyl)-2-thioxo-
Yield: 94%; m.p 178-180°C; Rf: 0.52 (ethyl aceta-
1,3,4-oxadiazol-3(2H)-yl)-1-phenylethanone (5)
te/hexanes, 2:8)1H-NMR (400 MHz, DMSO-d6):
8.22 (d,
Yield: 95%; m.p 128-130°C; Rf: 0.55 (ethyl aceta- 1H, J(19,22) = 1.6 Hz, H-19), 7.96 (dd, 1H, J(22,19) = 2.0 Hz
te/hexanes, 2:8); 1H-NMR (500 MHz, DMSO-d6):
8.01 (d, J(22,23) = 8.4 Hz, H-22), 7.84 (d, 1H, J(23,22) = 8.4 Hz, H-23),
2H, J(3,4),(5,4) = 7.5 Hz, H-3, H-5), 7.70 (t, 1H, J(10/9,11) = 7.5 7.52 (d, 2H, J(3,4),(5,4) = 8.0 Hz, H-3, H-5), 7.23 (t, 1H, J(10/9,11)
Hz, H-10) 7.57 (t, 2H, J(19/18,20), (21/20,22) = 7.5 Hz, H-19, H- = 8.0 Hz, H-10), 7.16 (d, 1H, J(11,10) = 7.6 Hz, H-11), 7.11 (s,
21), 7.52 (d, 2H, J(18,19),(22,21) = 7.5 Hz, H-18, H-22) 7.23 (t, 1H, NH), 7.07 (t, 1H, J(4/3,5) = 8.0 Hz, H-4), 6.83 (t, 1H,
1H, J(20/19,21) = 8.0 Hz, H-20) 7.15 (m, 1H, H-11), 7.14 (s, 1H, J(9/8,10) = 7.6 Hz, H-9), 6.19 (d, 1H, J (8,9) = 8.0 Hz, H-8), 5.03
684 Medicinal Chemistry, 2018, Vol. 14, No. 7 Shah et al.

(s, 2H, 16-CH2), 4.35 (s, 2H, 13-CH2); ESI-MS m/z: 539.0 1545 (C=N), 1540 (C=C), 1463 (C=S), 1165 (C-O); Anal.
(M+); HR-ESI-MS m/z Calcd for C23H15Cl4N3O2S [M] + m/z Calcd C24H18BrCl2N3O2S: C, 51.17; H, 3.22; Br, 14.19;
539.2611, Found 539.2610; IR (KBr, cm-1): 3265 (N-H), Cl, 12.59; N, 7.46; O, 5.68; S, 5.69 Found: C, 51.16; H, 3.21;
1629 (C=O), 1532 (C=N), 1576 (C=C), 1425 (C=S), 1070 N, 7.45.
(C-O); Anal. m/z Calcd for for for C23H15Cl4N3O2S: C,
51.23; H, 2.80; Cl, 26.30; N, 7.79; O, 5.93; S, 5.95 Found: C, 5.12. 2-(5-(2-(2,6-Dichlorophenylamino)benzyl)-2-thioxo-
51.23; H, 2.82; N, 7.78. 1,3,4-oxadiazol-3(2H)-yl)-1-(3-nitrophenyl)ethanone (12)

5.9. 3-(4-Chlorobenzyl)-5-(2-(2,6-dichlorophenylamino) Yield: 92%; m.p 123-125°C; Rf: 0.50 (ethyl aceta-
benzyl)-1,3,4-oxadiazole-2(3H)-thione (9) te/hexanes, 2:8); 1H-NMR (400 MHz, DMSO-d6):
8.70(s,
1H, H-23), 8.52 (d, 1H, J (21,20) = 8.4 Hz, H-21), 8.44 (d, 1H,
Yield: 90%; m.p 110-112°C; Rf: 0.51 (ethyl aceta- J(19,20) = 8.0 Hz, H-19), 7.88 (t, 1H, J(20/19,21) = 8.0 Hz, H-20),
te/hexanes, 2:8); 1H-NMR (500 MHz, DMSO-d6):
7.23 (d, 7.52 (d, 2H, J(3,4),(5,4) = 7.5 Hz, H-3, H-5), 7.23 t, 1H, J(10/9,11)
2H, J(3,4),(5,4) = 8.0 Hz, H-3, H-5), 7.19 (d, 2H, J(18,19),(21,22) = = 8.0 Hz, H-10), 7.16 (d, 1H, J (11,10) = 7.6 Hz, H-11 ), 7.12
8.0 Hz, H-18, H-21) 7.37 (t, 1H, J(20/21,22) = 7.5 Hz, H-20), (s, 1H, NH), 7.07 (t, 1H, J(4/3,5) = 8.0 Hz, H-4), 6.83 (t, 1H,
7.23 (m, 2H, H-10, H-11) 7.12 (s, 1H, NH), 7.08 (t, 1H, J(9/8,10) = 7.6 Hz, H-9), 6.19 (d, 1H, J (8,9) = 8.0 Hz, H-8) 5.14
J(4/3,5) = 8.0 Hz, H-4) 6.86 (t, 1H, J(9/8,10) = 7.5 Hz, H-9), 6.21 (s, 2H, 16-CH2), 4.35 (s, 2H, 13-CH2); ESI-MS m/z: 515.0
(d, 1H, J(8,9) = 7.5 Hz, H-8), 4.42 (s, 2H, 16-CH2), 4.35 (s,
(M+); HR-ESI-MS m/z Calcd for C23H16Cl2N4O4S [M] +
2H, 13-CH2); 13C-NMR: (75 MHz, DMSO-d6):
C, 166.4
515.3685, Found 515.3684; IR (KBr, cm-1): 3215 (N–H),
(C=S), 142.9 (C-14), 136.8 (C-2, C-6), 135.8 (C-20), 132.2
1692 (C=O), 1572 (C=N), 1579 (C=C), 1423 (C=S), 1041
(C-17), 131.5 (C-7), 130.7 (C-12), 130.5 (C-19, C-21), 129.1
(C-18, C-22), 128.4 (C-3, C-5), 128.1 (C-9), 126.4 (C-8), (C-O); Anal. m/z Calcd for for C23H16Cl2N4O4S: C, 53.60; H,
122.2 (C-9), 120.4 (C-10), 34.8 (13-CH2), 27.2 (16-CH2); 3.13; Cl, 13.76; N, 10.87; O, 12.42; S, 6.22, Found: C,
ESI-MS m/z: 476.0 (M+); HR-ESI-MS m/z Calcd for 53.62; H, 3.12; Cl, N, 10.88.
C22H16Cl3N3OS [M]+ 476.8059, Found 476.8058; IR (KBr,
cm-1): 3210 (N-H), 1546 (C=N), 1548 (C=C), 1436 (C=S), 5.13. 5-(2-(2,6-Dichlorophenylamino)benzyl)-3-(2-(4-ni-
1187 (C–O); Anal. m/z Calcd C22H16BrCl2N3OS: C, 55.42; trophenyl)-2-thioxoethyl)-1,3,4-oxadiazol-2(3H)-one (13)
H, 3.38; Cl, 22.31; N, 8.81; O, 3.36; S, 6.72 Found: C, 55.41; Yield: 90%; m.p 135-137°C; Rf: 0.49 (ethyl aceta-
H, 3.36; N, 8.80. te/hexanes, 2:8); 1H-NMR (300 MHz, DMSO-d6):
8.37 (d,
2H, J(20,19),(22,23) = 9.0 Hz, H-20, H-22), 8.24 (d, 2H,
5.10. 1-(4-Bromophenyl)-2-(5-(2-(2,6-dichlorophenylami- J(19,20),(23,22) = 8.7 Hz, H-19, H-23) 7.52 (d, 2H, J(3,4),(5,4) = 8.0
no)benzyl)-2-thioxo-1,3,4-oxadiazol-3(2H)-yl)ethanone
Hz, H-3, H-5) 7.24 (m, 2H, H-10, H-11), 7.11 (s, 1H, NH),
(10)
7.08 (t, 1H, J(4/3,5) = 6.9 Hz, H-4), 6.84 (t, 1H, J(9/8,10) = 7.5
Yield: 92%; m.p 148-150°C; Rf: 0.53 (ethyl aceta- Hz, H-9), 6.20 (d, 1H, J(8, 9) = 8.1 Hz, H-8), 5.10 (s, 2H, 16-
te/hexanes, 2:8); 1H-NMR (300 MHz, DMSO-d6):
7.95 (d, CH2), 4.35 (s, 2H, 13-CH2); ESI-MS m/z: 515.0 (M+); HR-
2H, J(23,22),(19,20) = 8.4 Hz, H-23, H-19), 7.78 (d, 2H, ESI-MS m/z Calcd for C23H16Cl2N4O4S [M]+ 515.3685,
J(20,19),(22,23) = 8.4 Hz, H-20, H-22), 7.53 (d, 2H, J(3,4),(5,4) = 8.1 Found 515.3684; IR (KBr, cm-1): 3215 (N-H), 1689 (C=O),
Hz, H-3, H-5), 7.24 (t, 1H, J(10/9,11) = 8.2 Hz, H-10), 7.16 (m, 1557 (C=N), 1545 (C=C),1434 (C=S), 1180 (C–O); Anal.
2H, H-11, NH), 7.08 (t, 1H, J(4/3,5) = 7.5 Hz, H-4), 6.84 (t, m/z Calcd for C23H16Cl2N4O4S: C, 53.60; H, 3.13; Cl, 13.76;
1H, J(9/8,10) = 7.2 Hz, H-9), 6.20 (d, 1H, J (8,9) = 8.0 Hz, H-8), N, 10.87; O, 12.42; S, 6.22, Found: C, 53.62; H, 3.11; N,
5.02 (s, 2H, 16-CH2), 4.35 (s, 2H, 13-CH2); ESI-MS m/z: 10.86.
549.0 (M+); HR-ESI-MS m/z Calcd for C23H16BrCl2N3O2S
[M]+ 549.2670, Found 549.2671; IR (KBr, cm-1): 3285 (N- 5.14. 2-(5-(2-(2,6-Dichlorophenylamino)benzyl)-2-thioxo-
H), 1689 (C=O), 1552 (C=N), 1568 (C=C), 1445 (C=S), 1,3,4-oxadiazol-3(2H)-yl)-1-(4-methoxyphenyl)ethanone
1090 (C-O); Anal. m/z Calcd for C23H16BrCl2N3O2S: C, (14)
50.29; H, 2.94; Br, 14.55; Cl, 12.91; N, 7.65; O, 5.83; S,
5.84 Found: C, 50.28; H, 2.92; N, 7.62. Yield: 91%; m.p 130-132°C; Rf: 0.51 (ethyl aceta-
te/hexanes, 2:8); 1H-NMR (300 MHz, DMSO-d6):
8.00 (d,
5.11. 1-(3-Bromophenyl)-3-(5-(2-(2,6-dichlorophenylami- 2H, J(19,20),(23,22) = 8.8 Hz, H-19, H-23), 7.53 (d, 2H,
no)benzyl)-2-thioxo-1,3,4-oxadiazol-3(2H)-yl)propan-1- J(20,21),(22,23) = 8.0 Hz, H-20, H-23), 7.23 (t, 1H, J(9/8,10) = 8.0
one (11) Hz, J(10/9,11) = 8.0 Hz, H-9, H-10), 7.16 (d, 2H, J(3,4),(5,4) = 8.4
Yield: 94%; m.p 134-136°C; Rf: 0.52 (ethyl aceta- Hz, H-3, H-5), 7.07 (d, 1H, J (11,10) = 7.5 Hz, H-11), 7.04 (s,
te/hexanes, 2:8); 1H-NMR (300 MHz, DMSO-d6):
8.15 (s, 1H, NH), 6.84 (t, 1H, J(4/3,5) = 6.9 Hz, H-4), 6.20 (d, 1H, J (8,9)
1H, H-23), 8.01 (d, 1H, J(21,20) = 9.8 Hz, H-21), 7.90 (d, 1H, = 8.0 Hz, H-8), 4.99 (s, 2H, 16-CH2), 4.35 (s, 2H, 13-CH2);
J(19,20) = 9.8 Hz, H-19), 7.55 (m, 3H, H-3, H-5, H-10), 7.24
13
C-NMR: (75 MHz, DMSO-d6):
C, 190.6 (C=O), 166.2
(t, 1H, J(20/19,21) = 8.1 Hz, H-20), 7.16 (m, 1H, H-10), 7.12 (s, (C=S), 163.6 (C-14), 162.9 (C-2, C-6), 142.8 (C-1), 136.9
1H, NH), 7.08 (t, 1H, J(4/3,5) = 7.5 Hz, H-4) 6.84(t, 1H, J(9/8,10) (C-19, C-23), 131.5 (C-20, C-22), 130.7 (C-3, C-5), 130.4
= 7.5 Hz, H-9), 6.21 (d, 1H, J(8,9) = 7.8 Hz, H-8), 5.04 (s, 2H, (C-7), 129.1 (C-18), 128.1 (C-21), 127.8 (C-12), 126.3 (C-
16-CH2), 4.35 (s, 2H, 13-CH2); ESI-MS m/z: 476.0 (M+); 4), 122.2 (C-8), 120.4 (C-9), 115.5 (C-10), 114.0 (C-4), 55.6
HR-ESI-MS m/z Calcd for C22H16Cl3N3OS [M] + 476.8059, (OCH3), 40.9 (13-CH2), 27.1 (C-16); ESI-MS m/z: 499.0
Found 476.8058; IR (KBr, cm-1): 3212 (N-H), 1650 (C=O), (M+); HR-ESI-MS m/z Calcd for C24H19Cl2N3O3S [M] +
Diclofenac 1,3,4-Oxadiazole Derivatives Medicinal Chemistry, 2018, Vol. 14, No. 7 685

499.0524, Found 499.0522; IR (KBr, cm-1): 3221 (N-H), C29H21Cl2N3O2S: C, 63.74; H, 3.87; Cl, 12.98; N, 7.69; O,
1678 (C=O), 1562 (C=N), 1528 (C=C),1423 (C=S), 1026 5.86; S, 5.87 Found: C, 63.73; H, 3.86; N, 7.68.
(C-O); Anal. m/z Calcd for for for C24H19Cl2N3O3S; C,
57.61; H, 3.83; Cl, 14.17; N, 8.40; O, 9.59; S, 6.41 Found: C, 5.18. 3-(2,4-Dichlorobenzyl)-5-(2-(2,6-dichlorophenylami-
57.60; H, 3.82; N, 8.41. no)benzyl)-1,3,4-oxadiazole-2(3H)-thione (18)
Yield: 92%; m.p 128-130°C; Rf: 0.50 (ethyl aceta-
5.15. 2-(5-(2-(2,5-Dichlorophenylamino)benzyl)-2-thioxo- te/hexanes, 2:8); 1H-NMR (500 MHz, DMSO-d6):
7.23 (d,
1,3,4-oxadiazol-3(2H)-yl)-1-(3-methoxyphenyl)ethan-1- 2H, J(3,4),(5,4) = 8.0 Hz, H-3, H-5), 7.19 (d, 2H, J(18,19),(21,22) =
one (15) 8.0 Hz, H-18, H-21) 7.37 (t, 1H, J(20/21,22) = 7.5 Hz, H-20),
Yield: 91 %; m.p 132-134°C; Rf: 0.48 (ethyl aceta- 7.23 (m, 2H, H-10, H-11) 7.12 (s, 1H, NH), 7.08 (t, 1H,
te/hexanes, 2:8); 1H-NMR (500 MHz, DMSO-d6):
7.61 (d, J(4/3,5) = 8.0 Hz, H-4) 6.86 (t, 1H, J(9/8,10) = 7.5 Hz, H-9), 6.21
1H, J(10,11) = 7.5 Hz, H-10), 7.52 (d, 2H, J(3,4),(5,4) = 8.0 Hz, H- (d, 1H, J(8,9) = 7.5 Hz, H-8), 4.42 (s, 2H, 16-CH2), 4.35 (s,
3, H-5), 7.48 (s, 1H, NH), 7.47 (d, 1H, J (19,20) = 8.0 Hz, H- 2H, 13-CH2); 13C-NMR: (75 MHz, DMSO-d6):
166.4
19), 7.27 (m, 2H, H-20, H-23), 7.16 (d, 2H, J (11,10) = J (21,20) = (C=S), 142.9 (C-14), 136.8 (C-2, C-6), 135.8 (C-20), 132.2
10.5 Hz, H-11, H-21) 7.06 (t, 1H, J(4/3,5) = 7.5 Hz, H-4), 6.83 (C-17), 131.5 (C-7), 130.7 (C-12), 130.5 (C-19, C-21), 129.1
(t, 1H, J(9/8,10) = 7.5 Hz, H-9), 6.20 (d, 1H, J (8,9) = 8.0 Hz, H- (C-18, C-22), 128.4 (C-3, C-5), 128.1 (C-9), 126.4 (C-8),
8) 5.03 (s, 2H, 16-CH2), 4.35 (s, 2H, 13-CH2), 3.81 (s, 3H, 122.2(C-9), 120.4 (C-10), 34.8 (13-CH2), 27.2 (16-CH2);
OCH3); ESI-MS m/z: 500.0 (M+); HR-ESI-MS m/z Calcd for ESI-MS m/z: 511.0 (M+); HR-ESI-MS m/z Calcd for
C24H19Cl2N3O3S [M]+ 500.3970, Found 500.3971; IR (KBr, C22H15Cl4N3OS [M]+ 511.251, Found 511.250; IR (KBr, cm-
1
cm-1): 3315 (N-H), 1691 (C=O), 1579 (C=N), 1576 ): 3215 (N-H), 1546 (C=N), 1549 (C=C), 1431 (C=S), 1185
(C=C),1443 (C=S), 1049 (C-O); Anal. m/z Calcd for (C-O); Anal. m/z Calcd C22H15Cl4N3OS: C, 51.68; H, 2.96;
C24H19Cl2N3O3S: C, 57.61; H, 3.83; Cl, 14.17; N, 8.40; O, Cl, 27.74; N, 8.22; O, 3.13; S, 6.27 2 Found: C, 51.66; H,
9.59; S, 6.41, Found: C, 57.62; H, 3.82; Cl, 14.15; N, 8.41. 2.94; N, 8.20.

5.16. 2-(5-(2-(2,6-Dichlorophenylamino)benzyl)-2-thioxo- 5.19. 2,6-Dichloro-N-(2-((5-(2,6-dichlorobenzylthio)-1,3,4-

1,3,4-oxadiazol-3(2H)-yl)-1-(2-hydroxyphenyl)ethanone oxadiazol-2-yl)methyl)phenyl)aniline (19)
(16) Yield: 96%; m.p 132-134°C; Rf: 0.50 (ethyl aceta-
Yield: 94%; m.p 164-166°C; Rf: 0.47 (ethyl aceta- te/hexanes, 2:8) 1H-NMR (500 MHz, DMSO-d6):
7.36 (d,
te/hexanes, 2:8); 1H-NMR (300 MHz, DMSO-d6):
11.13 (s, 2H, J(3,4),(5,4) = 7.5 Hz, H-3, H-5), 7.34 (d, 2H, J(19,20),(21,20) =
1H, OH), 7.81 (dd, 1H, J(23,22) = 2.4 Hz J(23,21) = 6.3 Hz, H- 8.0 Hz, H-19, H-21) 7.37 (t, 1H, J(20/21,22) = 7.5 Hz, H-20),
23), 7.53 (d, 3H, J(3,4),(5,4), (21,20) = 8.1 Hz, H-3, H-5, H-21), 7.23 (m, 2H, H-10, H-11), 7.12 (s, 1H, NH), 7.08 (t, 1H,
7.18 (m, 2H, H-10, H-11), 7.05 (d, 1H, J (22,23) = 7.2 Hz, H- J(4/3,5) = 8.0 Hz, H-4) 6.86 (t, 1H, J(9/8,10) = 7.5 Hz, H-9), 6.21
(d, 1H, J(22, 21) = 8.0 Hz, H-22), 4.62 (s, 2H, 16-CH2), 4.39 (s,
22), 7.01 (s, 1H, NH), 6.97 (t, 1H, J(4/3,5) = 7.8 Hz, H-4), 6.85
2H, 13-CH2); ESI-MS m/z: 511.9 (M+); HR-ESI-MS m/z
(t, 1H, J(9/8,10) = 8.1 Hz, H-9), 6.19 (d, 1H, J (8,9) = 7.5 Hz, H-
Calcd for C22H15Cl4N3OS [M]+ 511.2510, Found 511.2511;
8), 4.97 (s, 2H, 16-CH2), 4.35 (s, 2H, 13-CH2); ESI-MS m/z: IR (KBr, cm-1): 3217 (N-H), 1540 (C=N), 1544 (C=C),1430
486.0 (M+); HR-ESI-MS m/z Calcd for C23H17Cl2N3O3S (C=S), 1183 (C-O); Anal. m/z Calcd C22H16BrCl2N3OS: C,
[M]+ 486.3704, Found 486.3703; IR (KBr, cm-1): 3215 (N- 51.68; H, 2.96; Cl, 27.74; N, 8.22; O, 3.13; S, 6.27, Found:
H), 1692 (C=O), 1572 (C=N), 1579 (C=C),1423 (C=S), 1041 C, 51.67; H, 2.97; N, 8.21.
(C–O); Anal. m/z Calcd for for C23H17Cl2N3O3S : C, 56.80;
H, 3.52; Cl, 14.58; N, 8.64; O, 9.87; S, 6.59 Found: C, 56.82; 5.20. N-(2-((5-(2-Bromobenzylthio)-1,3,4-oxadiazol-2-
H, 3.51; N, 8.63. yl)methyl)phenyl)-2,6-dichloroaniline (20)

5.17. 1-(Biphenyl-4-yl)-2-(5-(2-(phenylamino)benzyl)-2- Yield: 93%; m.p 124-126°C; Rf: 0.52 (ethyl aceta-
thioxo-1,3,4-oxadiazol-3(2H)-yl)ethanone (17) te/hexanes, 2:8) 1H-NMR (400 MHz, DMSO-d6):
7.63 (d,
1H, J(19, 20) = 8.4 Hz, H-19), 7.52 (d, 2H, J(3,4),(5,4) = 8.0 Hz,
Yield: 90%; m.p 160-162°C; Rf: 0.56 (ethyl aceta- H-3, H-5) 7.47 (dd, 1H, J(8,11) = 1.3 Hz, J(8,9) = 6.0 Hz, H-8),
te/hexanes, 2:8): 1H-NMR (400 MHz, DMSO-d6):
8.08 (d, 7.27 (m, 4H, H-10, H-11, H-20, H-21) 7.13 (s, 1H, NH),
2H, J(19,20),(23,22) = 8.4 Hz, H-19, H-23), 7.87 (d, 2H, 7.50), 7.09 (t, 1H, J(4/3,5) = 8.0 Hz, H-4) 6.86 (t, 1H, J(9/8,10) =
J(20,19),(22,23) = 8.0 Hz, H-20, H-22), 7.77 (d, 2H, J(25,26),(29,28) = 8.0 Hz, H-9), 6.21 (d, 1H, J(22, 21) = 8.1 Hz, H-22), 4.51 (s,
7.6 Hz, H-25, H-29), 7.35( d, 4H, J(20,19),(22,23) = 8.0 Hz, H-20, 2H, 16-CH2), 4.37 (s, 2H, 13-CH2); 13C-NMR: (100.0 MHz,
H-22), 7.53 (m, 4H, H-3, H-5, H-23, H-28), 7.45 (t, 1H, DMSO-d6):
166.6 (C=S), 162.2 (C-14), 142.9 (C-2, C-6),
J(10/9,11) = 8.0 Hz, H-10), 7.23 (t, 1H, J(27/26,28) = 8.0 Hz, H- 136.8 (C-17), 135.3 (C-18), 132.8 (C-7), 131.5 (C-12), 131.4
27), 7.17 (m, 2H, H-11, NH), 7.07 (t, 1H, J(4/3,5)= 8.0 Hz, H- (C-21), 130.6 (C-22), 130.0 (C-3, C-5), 129.1 (C-19), 128.2
4), 6.84 (t, 1H, J(9/8,10) = 8.0 Hz, H-9), 6.20 (d, 1H, J (8,9) = 8.0 (C-20), 127.9 (C-11), 126.4 (C-8), 123.9 (C-9), 122.1 (C-
Hz, H-8), 5.08 (s, 2H, 16-CH2), 4.36 (s, 2H, 13-CH2); ESI- 10), 120.4 (C-1), 115.5 (C-4), 36.6 (13-CH2), 27.2 (16-CH2);
MS m/z: 546.0 (M+); HR-ESI-MS m/z Calcd for ESI-MS m/z: 519.0 (M+); HR-ESI-MS m/z Calcd for
C29H21Cl2N3O2S [M]+ 546.4669, Found 546.4668; IR (KBr, C22H16BrCl2N3OS [M]+ 521.2569, Found 521.2568; IR (K-
cm-1): 3281 (N-H), 1688 (C=O), 1562 (C=N), 1598 Br, cm-1): 3219 (N-H), 1542 (C=N), 1540 (C=C),1438
(C=C),1423 (C=S), 1086 (C-O); Anal. m/z Calcd for for for (C=S), 1181 (C-O); Anal. m/z Calcd C22H16BrCl2N3OS: C,
686 Medicinal Chemistry, 2018, Vol. 14, No. 7 Shah et al.

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Bioorg. Med. Chem., 2014, 22, 2855-2866.
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Ismail, Z.; Ahmad, A. Synthesis and immunomodulatory properties
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CONFLICT OF INTEREST [17] Mesaik, M.A.; Khan, K.M.; Rahat, S.; Zia-Ullah.; Choudhary, M.I.;
Murad, S.; Siddiqui, R.A. Immunomodulatory properties of syn-
The authors declare no conflict of interest, financial or thetic imidazolone derivatives. Lett. Drug Des. Discov., 2005, 2,
otherwise. 490-496.
[18] Khan, K.M.; Ziaullah.; Lodhi, M.A.; Jalil, S.; Choudhary, M.I.;
Atta-ur-Rahman. Synthesis and anti-inflammatory activity of some
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21, 139-143.
This work was supported by the Higher Education Com- [19] Choudhary, M.I.; Jalil, S.; Nawaz, S.A.; Khan, K.M.; Tareen, R.B.
mission (HEC) Pakistan (Project No. 20-1910), under the Antiinflammatory and lipoxygenase inhibitory compounds from
National Research Program for Universities. vitex agnus
castus. Phytother. Res., 2009, 23 1336-1339.
[20] Khan, K.M.; Ambreen, N.; Mughal, U.R.; Jalil, S.; Perveen, S.
Choudhary, M.I. 3-Formylchromones: potential antiinflammatory
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