Synthesis of tert-butyl (substituted benzamido)phenylcarbamate derivatives: anti-inflammatory activity and docking studies

Article abstract of DOI:10.1007/s12154-017-0168-x

A series of new tert-butyl 2-(substituted benzamido) phenylcarbamate (4a–4j) were synthesized by the condensation of tert-butyl 2-amino phenylcarbamate (3) with various substituted carboxylic acid in the presence of EDCI and HOBt as coupling reagent, obta

Full text of DOI:10.1007/s12154-017-0168-x

J Chem Biol
DOI 10.1007/s12154-017-0168-x
ORIGINAL ARTICLE
Synthesis of tert-butyl (substituted benzamido)phenylcarbamate
derivatives: anti-inflammatory activity and docking studies
1
1
1
1
Shankar Bhookya & Jalapathi Pochampally & Anil Valeru & Vianala Sunitha
&
2
2
3
Saikrishna Balabadra & Vijjulatha Manga & Karunakar rao Kudle
Received: 8 January 2017 /Accepted: 14 March 2017
#
Springer-Verlag Berlin Heidelberg 2017
Abstract A series of new tert-butyl 2-(substituted
benzamido) phenylcarbamate (4a–4j) were synthesized by
the condensation of tert-butyl 2-amino phenylcarbamate (3)
with various substituted carboxylic acid in the presence of
EDCI and HOBt as coupling reagent, obtain in excellent
yields. The structures of all newly synthesized compounds
were characterized spectroscopically and evaluated for
in vivo anti-inflammatory activity compared to the standard
drug, indomethacin, by using the carrageenan-induced rat paw
edema protocol. Most of the compounds exhibited a promis-
ing anti-inflammatory activity within 9 to 12 h, the percentage
of inhibition values ranging from 54.239 to 39.021%. The
results revealed that the compounds 4i and 4a exhibited better
or equivalent anti-inflammatory activity with the percentage
of inhibition of 54.239 and 54.130%, respectively, which was
comparable to standard drug. In addition to experimental re-
sults, in silico docking studies was used as a tool to verify and
expand the experimental outcomes.
Introduction
Inflammation is a key area of research for various pharmaceu-
tical companies. Arthritis, asthma, allergy, multiple sclerosis,
and additional diseases which cause inflammation and pain
are widely prevalent throughout the world. For the treatment
of pain and inflammation, various drugs such as indometha-
cin, ibuprofen, aspirin, nimisulide, celecoxib, rofecoxib, and
dichlofenac are available in the market [1, 2]. But long-term
use of these drugs causes various side effects such as ulcera-
tion, renal failure, gastrointestinal bleeding, and heart stroke
[3–5]. All these indicate that there is a need for safer anti-
inflammatory drugs. These serious side effects are limiting
the use of non steroidal anti-inflammatory drugs (NSAIDs)
in common inflammation cases. Interestingly, the replacement
of the carboxylic groups by amide groups in NSAID drugs
such as indomethacin, meclofenamic acid, and ketoprofen
conferred the compound’s greater selectivity for
cyclooxygenase-2 over the cyclooxygenase-1 enzyme [6, 7].
This enzyme exists in the form of COX-1, which plays a
crucial cytoprotective role in the gastrointestinal tract, whereas
the second form COX-2 is responsible for the production of
PGs during inflammation [8]. The inhibition of prostanoids
was produced by COX-2 which might be decided to the an-
ti-inflammatory, analgesic, and antipyretic affects of NSAIDs
.
.
Keywords Amide derivatives Anti-inflammatory activity
.
Paw edema Docking studies
Electronic supplementary material The online version of this article
doi:10.1007/s12154-017-0168-x) contains supplementary material,
which is available to authorized users.
(
[9]. Therefore, development of novel compounds with anti
*
Jalapathi Pochampally
pochampalli.ou.chemi@gmail.com
inflammatory agents like an alternative to NSAIDs and im-
proved safety profile is of believable importance.
Amide formation from carboxylic acids and amines is a
fundamental reaction in organic, biological, medicinal, poly-
mer, and material chemistry for which a great amount of re-
search is still pursued [10]. The amide derivatives were asso-
ciated with a broad spectrum of biological activities including
anti-inflammatory [11], antidegenerative [12], antiplatelet
[13], anticancer [14, 15], antimicrobial [16–18], urokinase
1
2
3
Department of Chemistry, University College of Science, Osmania
University, Saifabad, Hyderabad 500004, India
MMMC Group, Department of Chemistry, University College of
Science, Osmania University, Hyderabad, Telangana 500007, India
Department of Biochemistry, University College of Science,
Osmania University, Hyderabad, Telangana 500007, India

J Chem Biol
inhibitor [19], antituberculosis [20, 21], anticonvulsant [22],
insecticide [23], and antitumor [24]. And amide derivatives
possessing anti-inflammatory activities have been reported
in the literature [25–27]. Encouraged by these results, it was
attempted in the present study to synthesize a novel series of
tert-butyl 2-(substituted benzamido) phenylcarbamate ana-
logues (4a–4j) and screening for their anti-inflammatory ac-
tivity. The results revealed that several derivatives showed
promising activities. The in silico docking study of all the
compounds was also performed to provide the binding modes
of COX-2 enzyme.
J = 6.76, 2.14 Hz), 6.30 (s, 2H, NH ), 1.38 (s, 9H, 3CH ) ppm;
2
3
1
3
C NMR (100 MHz, DMSO-d ): δ = 152.5, 141.5, 125.5,
6
125.1, 122.8, 118.9, 114.5, 79.5, 28.4 ppm; IR (KBr): νꢀ
−
1
= 3355.7, 3100.4, 1685.3, 1642.2 cm ; MS (ESI+):
+
m/z = 209.15 ([M + H] ).
Spectroscopic data of compounds 4a–4j
General procedure for the synthesis of compounds 4a–4j to a
stirred solution of 0.150 g benzoic acid (1.07 mmol, 1 eq), in a
3
5 cm N,N-dimethylformamide at cooling at 0 °C then it was
3
added 0.153 cm of DIPEA (N,N-diisopropylethylamine)
(1.605 mmol, 1.5 eq), 0.244 g of compound 3 (1.177 mmol,
Materials and methods
Chemistry
1.1 eq) and 0.306 g EDCI (1-(3-dimethyl aminopropyl)-3-
ethylcarbodiimide.HCl) (1.605 mmol, 1.5 eq) and 0.216 g of
HOBt (hydroxybenzotriazole) (1.605 mmol, 1.5 eq) succes-
sively. The reaction mixture was stirred for 30 min and it was
kept at room temperature for 3 h to complete the reaction. The
completion of the reaction was monitored by TLC. The crude
product was diluted with water, extracted with diethyl ether,
and separated the organic layer, washed with water and brine
solution. The organic layer was dried over anhydrous Na SO
All starting materials, reagents, and solvents were commer-
cially available and used after purification. All the melting
points are uncorrected and were determined in open capillary
tubes using sulfuric acid bath. IR spectra were recorded on
1
13
Perkin-Elmer 1000 instrument using KBr pellet. H and
C
2
4
NMR spectra were obtained in CDCl calibrated solvents on a
and concentrated, to obtain crude solid which was purified by
column chromatography eluting with 20–40% ethyl acetate in
hexane, to obtain corresponding desired compounds 4a–4j as
solids.
3
VARIAN spectrometer at 500 and 125 MHz, respectively.
Chemical shift signals are given in δ (parts per million) rela-
tive to TMS, and coupling constants (J) are expressed in Hertz
(
Hz). Combinations of the following abbreviations are used to
describe NMR spectra: s (singlet), d (doublet), t (triplet) q
quartet), and m (multiplet). Flash column chromatography
Preparation of tert-butyl 2-(4-fluorobenzamido)
phenylcarbamate (4a)
(
was performed using silica gel (Merck, 60–120 Mesh).
Commercially available reagents were used as supplied and
some of them were distilled before use. All reactions were
performed in oven-dried glassware. Electron Spray
Ionization (ESI) and high-resolution mass spectra were re-
corded on a QSTARXL hybrid MS/MS system (Applied Bio
systems, USA) under electrospray ionization.
From the 0.150 g 4-florobenzoic acid (1.07 mmol, 1 eq),
0.153 cm DIPEA (1.605 mmol, 1.5 eq), 0.244 g compound
3
3 (1.177 mmol, 1.1 eq), 0.306 g EDCI (1.605 mmol, 1.5 eq),
and 0.216 g HOBt (1.605 mmol, 1.5 eq), the compound 4a
was obtained as white-colored solid (0.275 g, 74%) after chro-
matography on a silica gel column with petroleum ether/ethyl
1
acetate (80:20, v/v). m.p. 250–252 °C; H NMR (400 MHz,
Preparation of tert-butyl 2-amino phenyl carbamate (3)
CDCl ): δ = 9.48 (s, 1H, NH), 8.65 (s, 1H, NH), 7.94 (d, 2H,
3
J = 8.25 Hz, Ar-H), 7.75 (d, 2H, J = 7.98 Hz, Ar-H), 7.36–
1
3
A mixture of 0.800-g compound 2 (3.361 mmol, 1.0 eq) and
7.26 (m, 4H, Ar-H), 1.48 (s, 9H, 3CH ) ppm; C NMR
3
0
.779 g FeCl (3.361 mmol, 1.0 eq) was dissolved in methanol
(100 MHz, DMSO-d ): δ = 165.92, 164.66, 163.6, 154.65,
130.50, 130.02, 129.70 (2C), 125.93(2C), 124.38 (2C),
3
6
3
3
(
10 cm ), and N H ·H O (5 cm ) were added at room temper-
2
4
2
ature. The reaction mixture was heated under reflux for 3 h.
The completion of the reaction was monitored by TLC. The
contents were cooled to room temperature, concentrated under
reduced pressure to remove the solvent. Then, it was basified
115.5 (2C), 81.20, 28.19 (3C) ppm; IR (KBr): νꢀ = 3266.7,
−
1
3064.2, 1693.5, 1650.4, 752.3 cm ; MS (ESI+):
+
m/z = 331.05 ([M + H] ), 348.07, 353.02; anal calcd for:
C H N O : C, 70.73; H, 9.81; N, 7.17; O, 12.29; found:
2
3 38 2 3
with saturated NaHCO solution and extracted with DCM; it
C, 70.78; H, 9.83; N, 7.20; O, 12.33.
3
was washed with water and brine. The organic layer was dried
over Na SO and concentrated to obtained crude solid which
was purified by column chromatography eluting with 20%
tert-butyl 2-(4-methylbenzamido)phenylcarbamate (4b)
From the 0.150 g 4-methyle benzoic acid (1.1 mmol, 1 eq),
2
4
ethyl acetate in hexane to produce compound 3 as white solid.
1
3
H NMR (400 MHz, CDCl ): δ = 9.25 (s, 1H), 8.00 (d, 1H,
0.157 cm DIPEA (1.65 mmol, 1.5 eq), 0.251 g compound 3
3
J = 1.06 Hz); 7.40 (dd, 1H, J = 6.80, 2.14 Hz); 6.79 (dd, 1H,
(1.21 mmol, 1.1 eq), 0.315 g EDCI (1.65 mmol, 1.5 eq) and

J Chem Biol
+
0
.222 g HOBt (1.65 mmol, 1.5 eq) the compound 4b was
MS (ESI+): m/z = 438.95.([M + H] ), 455.97; anal calcd for:
obtained as off white colored solid (0.210 g, 55.5%) after
chromatography on a silica gel column with petroleum
ether/ethyl acetate (80:20, v/v). m.p. 210–212 °C; H NMR
C H IN O : C, 49.33; H, 4.37; I, 28.96; N, 6.39; O, 10.95;
1
8
19
2 3
found: C, 49.30; H, 4.38; I, 28.94; N, 6.36; O, 10.94.
tert-butyl 2-(3,5-dinitrobenzamido)phenylcarbamate (4e)
From the 0.100 g 3,5-dinitrobenzoic acid (0.47 mmol, 1 eq),
1
(
400 MHz, CDCl ): δ = 9.80 (s, 1H, NH), 9.01 (s, 1H, NH),
3
7
.87 (d, 2H, J = 8.00 Hz, Ar-H), 7.35 (s, 2H, J = 7.93 Hz, Ar-
H), 7.28 (d, 2H, J = 8.05 Hz, Ar-H), 7.21 (d, 2H, J = 7.55 Hz,
1
3
3
Ar-H), 2.51 (s, 3H, CH ), 1.45 (s, 9H, 3CH ) ppm; C NMR
0.089 cm DIPEA (0.705 mmol, 1.5 eq), 0.107 g compound 3
3
3
(
100 MHz, DMSO-d ): δ = 165.73, 154.55, 152.23, 142.33,
31.24, 130.05, 129.16, 127.37, 125.80(2C), 124.46, 81.06,
(0.517 mmol, 1.1 eq), 0.134 g EDCI (0.705 mmol, 1.5 eq) and
0.095 g HOBt (0.705 mmol, 1.5 eq) the compound 4e was
obtained as brick red-colored solid (0.140 g, 67.6%) after
chromatography on a silica gel column with petroleum
6
1
2
2
8.22(3C), 21.45 ppm; IR (KBr): νꢀ = 3332.4, 3249.1, 3069.9,
−
1
908.8, 1799.6, 1690.8 cm ; MS (ESI+): m/z = 327.08
+
1
(
[M + H] ), 349.04; anal calcd for: C H N O : C, 69.92,
ether/ethyl acetate (80:20, v/v). m.p. 248–250 °C; H NMR
1
9 22 2 3
H, 6.79, N, 8.58, O, 14.71; found: C, 69.42, H, 6.38, N, 8.52,
O, 14.75.
(400 MHz, CDCl ): δ = 10.02 (s, 1H, NH), 9.42 (s, 1H, Ar-H),
3
8.19 (s, 1H, NH), 8.98 (d, 2H, J = 7.96 Hz, Ar-H), 7.79 (d, 2H,
J = 8.0 Hz, Ar-H), 7.40 (d, 2H, J = 8.5 Hz, Ar-H), 1.50 (s, 9H,
1
3 6
3
tert-butyl 2-(4-tert-butylbenzamido)phenylcarbamate (4c)
3CH ) ppm; C NMR (100 MHz, DMSO-d ): δ = 160.93,
1
53.58, 147.98, 137.33, 131.55(2C), 127.71(2C), 126.28,
From the 0.150 g 4-tert-butylbenzoic acid (0.84 mmol, 1 eq),
127.38(2C), 123.89, 123.44, 120.53, 80.25, 27.8(3C) ppm;
3
−1
0
.120 cm DIPEA (1.26 mmol, 1.5 eq), 0.192 g compound 3
IR (KBr): νꢀ = 3222.5, 2826.1, 1772.6, 1630.5 cm ; MS
+
(
0
0.924 mmol, 1.1 eq), 0.240 g EDCI (1.26 mmol, 1.5 eq) and
.170 g HOBt (1.26 mmol, 1.5 eq) the compound 4c was
(ESI+): m/z = 403.00 ([M + H] ), 425.00; anal calcd for:
C H N O : C, 53.43; H, 4.51; N, 13.92; O, 27.83; found:
1
8 18 4 7
obtained as off white colored solid (0.285 g, 83.3%) after
chromatography on a silica gel column with petroleum
ether/ethyl acetate (70:30, v/v). m.p. 215–217 °C; H NMR
C, 53.73; H, 4.52; N, 13.90; O, 27.84.
1
tert-butyl 2-dodecanamidophenylcarbamate (4f)
From the 0.150 g dodecanoic acid (0.75 mmol, 1 eq),
(
400 MHz, CDCl ): δ = 9.38 (s,1H, NH), 8.64 (s, 1H, NH),
3
7
.99 (d, 2H, J = 7.85 Hz, Ar-H), 7.30 (d, 2H, J = 8.01 Hz, Ar-
3
H), 7.21 (m, 2H, Ar-H), 7.10 (d, 2H, J = 7.01 Hz, Ar-H), 1.55
0.107 cm DIPEA (1.125 mmol, 1.5 eq), 0.175 g compound
1
3
(
s, 9H, 3CH ), 1.28 (s, 9H, 3CH ) ppm; C NMR (100 MHz,
3 (0.825 mmol, 1.1 eq), 0.214 g EDCI (1.125 mmol, 1.5 eq)
and 0.151 g HOBt (1.125 mmol, 1.5 eq) the compound 4f was
obtained as white-colored solid (0.240 g, 73.8%) after chro-
matography on a silica gel column with petroleum ether/ethyl
3
3
DMSO-d ): δ = 165.54, 157.26, 155.13, 149.70, 130.14,
6
1
8
=
28.54, 127.68(2C), 126.55(2C), 124.70(2C), 124.23(2C),
0.05, 34.03, 30.19(3C). 27.42(3C) ppm; IR (KBr): νꢀ
−1
1
3266.9, 3064.2, 1693.5, 1650.5, 1442.8 cm ; MS (ESI+):
acetate (80:20, v/v). m.p.: 260–262 °C; H NMR (400 MHz,
+
m/z = 369.12 ([M + H] ), 391.16; anal calcd for: C H N O :
CDCl ): δ = 9.20(s, 1H, NH), 8.04(s, 1H, NH), 7.39 (d, 2H,
2
2
28
2
3
3
C, 71.71, H, 7.66, N, 7.60, O, 13.03; found: C, 71.74; H, 7.68;
N, 7.60; O, 13.05.
J = 7.38 Hz, Ar-H), 7.14 (m, 2H, Ar-H), 2.36 (t, 2H, CH2),
1.67 (t, 2H, CH ), 1.52 (s, 9H, 3CH ), 1.28 (s, 16H, CH ),
2
3
2
1
3
0
.88 (t, 3H, CH ) ppm; C NMR (100 MHz, DMSO-d6):
3
tert-butyl 2-(2-iodobenzamido)phenylcarbamate (4d)
δ = 178.82, 172.65, 154.27, 130.68, 129.91, 125.24(2C),
24.44(2C), 80.80, 37.21, 31.83, 29.52(3c), 29.26, 29.02,
1
From the 0.150 g 2-iodobutylbenzoic acid (0.600 mmol, 1 eq),
28.23(3C), 25.72, 24.68, 14.03 ppm; IR (KBr): νꢀ = 3420.3,
3
−1
0
.085 cm DIPEA (0.900 mmol, 1.5 eq), 0.137 g compound 3
3380.6, 1680.6, 1648.2 cm ; MS (ESI+): m/z = 391.05
+
(
0
0.66 mmol, 1.1 eq), 0.171 g EDCI (0.9 mmol, 1.5 eq) and
.121 g HOBt (0.90 mmol, 1.5 eq) the compound 4d was
([M + H] ), 413.2; anal. calcd for: C H N O : C, 70.73,
2
3 38 2 3
H, 9.81, N, 7.17; O, 12.29. Found: C, 70.72, H, 9.84, N,
7.18, O, 12.29.
obtained as yellow colored solid (0.195 g, 67.9%) after chro-
matography on a silica gel column with petroleum ether/ethyl
1
acetate (80:20, v/v). m.p. 225–227 °C; H NMR (400 MHz,
tert-butyl 2-(4-chloro-2,5-difluorobenzamido)
phenylcarbamate(4 g)
CDCl ): δ = 9.16 (s, 1H, NH), 8.26 (s, 1H, NH), 7.91 (d, 1H,
3
J = 7.93 Hz, Ar-H), 7.64 (d, 1H, J = 7.17 Hz, Ar-H), 7.53–
7
3
1
1
.38 (m, 3H, Ar-H), 7.24–7.12 (m, 3H, Ar-H), 1.47(s, 9H,
From the 0.100 g 4-chloro-2,5-difluorobenzoic acid
(0.520 mmol, 1 eq), 0.074 cm DIPEA (0.78 mmol, 1.5 eq),
0.118 g compound 3 (1.572 mmol, 1.1 eq), 0.148 g EDCI
(0.78 mmol, 1.5 eq) and 0.105 g HOBt (0.78 mmol, 1.5 eq)
the compound 4 g was obtained as white colored solid
1
3
3
CH ) ppm; C NMR (100 MHz, DMSO-d ): δ = 168.10,
3
6
53.96, 152.12, 141.63, 139.92, 131.34, 131.26, 128.93,
28.08, 126.70(2C), 125.25(2C), 92.55, 80.88, 28.25 ppm;
−1
IR (KBr): νꢀ = 3341.2, 3246.4, 1723.8, 1648.5, 748.7 cm ;

J Chem Biol
(
0.190 g, 95.0%) after chromatography on a silica gel column
80.85, 36.38, 28.21(3C), 25.83, 24.24 ppm; IR (KBr): νꢀ
−1
with petroleum ether/ethyl acetate (70:30, v/v). m.p.: 238–
2
8
= 3074.2, 2926.9, 2855.1, 1743.39, 1665.0 cm ; MS (ESI+
1
+
40 °C; H NMR (400 MHz, CDCl ): δ = 9.59 (s, 1H, NH),
): m/z = 394.22 ([M + H] ), 416.26; anal calcd for:
3
.98 (s, 1H, NH), 8.0(t, 1H, Ar-H), 7.71 (d, 1H, J = 8.24 Hz,
C H N O : C, 70.21; H, 6.92; N, 10.68; O, 12.20; found:
2
3 27 3 3
1
3
Ar-H), 7.39–7.20 (m, 3H, Ar-H), 1.51 (s, 9H, 3CH ) ppm; C
C, 70.22; H, 6.94; N, 10.70; O, 12.20.
3
NMR (100 MHz, DMSO-d ): δ = 168.10, 158.55, 153.96,
6
1
1
50.96, 141.63, 139.92, 128.93, 128.08, 126.79,
25.27(2C), 119.24, 115.10, 80.88, 28.25(3C) ppm; IR
tert-butyl 2-(2-bromoacetamido)phenylcarbamate (4j)
From the 0.100 g 2-bromoacetic acid (0.72 mmol, 1 eq),
−
1
(
(
KBr): νꢀ = 34O2.4, 3305.0, 1713.3, 1665.0 cm ; MS
+
3
ESI+): m/z = 383.01 ([M + H] ), 400.03; anal. calcd for:
0.140 cm DIPEA (1.08 mmol, 1.5 eq), 0.164 g compound
C H ClF N O C, 56.48; H, 4.48; Cl, 9.26; F, 9.93; N,
3 (0.79 mmol, 1.1 eq), 0.206 g EDCI (1.08 mmol, 1.5 eq) and
0.145 g HOBt (1.08 mmol, 1.5 eq) the compound 4j was
obtained as brick red-colored solid (0.250 g, 75.7%) after
chromatography on a silica gel column with petroleum
1
8
17
2 2 3:
7
.32; O, 12.54; found: C, 56.45; H, 4.49; Cl, 9.22; F, 9.90;
N, 7.25; O, 12.42.
1
tert-butyl 2-(2,4,5-trimethoxybenzamido)phenylcarbamate
ether/ethyl acetate (80:20, v/v). m.p.: 180–182 °C; H NMR
(
4 h)
(400 MHz, CDCl ): δ = 3402, 3305, 2972.6, 1717.8, 1658.2,
3
1
7
68.5. H–NMR- (500 MHz) (CDCl3)-d : δ 9.64 (s, 1H, NH),
6
From the 0.100 g 4-chloro-2,5-difluorobenzoic acid
8.99 (s, 1H, NH), 8.89 (d, 2H, J = 8.0 Hz, Ar-H), 7.20–7.1.52
(m, 2H, Ar-H), 3.62 (s, 2H, −CH -), 1.45 (s, 9H, 3CH ) ppm;
3
(
0.47 mmol, 1 eq), 0.090 cm DIPEA (0.705 mmol, 1.5 eq),
.107 g compound 3 (0.512 mmol, 1.1 eq), 0.134 g EDCI
0.705 mmol, 1.5 eq) and 0.095 g HOBt (0.705 mmol,
.5 eq) the compound 4 h was obtained as white colored solid
0.150 g, 72.4%) after chromatography on a silica gel column
with petroleum ether/ethyl acetate (80:20, v/v). m.p.: 188–
2
3
1
3
0
(
1
(
C NMR (100 MHz, DMSO-d ): δ = 171.45, 154.11, 130.76,
6
129.77, 126.20(2C), 125.29 (2C), 80.05, 28.24(3C),
24.08 ppm; IR (KBr): νꢀ = 3074.2, 2926.9, 2855.1, 1743.39,
−1
+
1665.0 cm ; MS (ESI+): m/z = 643.22 ([M + H] ); anal calcd
for: C H Br O : C, 47.43; H, 5.21; Br, 24.27; N, 8.51; O,
2
5
32
2 6
1
1
8
90 °C; H NMR (400 MHz, CDCl ): δ = 9.02 (s, 1H, NH),
14.58; found: C, 47.61; H, 5.25, Br, 24.32; N, 8.47; O, 14.65.
3
.40 (s, 1H, NH), 7.9 (d, 2H, J = 8.6 Hz, Ar-H), 7.3 (s, 1H, Ar-
H), 7.18 (d, 2H, J = 8.6 Hz), 6.92 (s, 1H, Ar-H), 3.68 (s, 9H,
Carrageenan-induced hind paw edema test
1
3
3
O–CH ), 1.51 (s, 9H, 3CH ) ppm; C NMR (100 MHz,
3 3
DMSO-d ): δ = 163.59, 153.77, 152.88, 152.62, 143.3,
In this study, the in vivo anti-inflammatory activity was eval-
uated for all the newly synthesized compounds 4a–4j using
the carrageenan-induced rat paw edema method. For the de-
termination of anti-inflammatory effect, the carrageenan-
induced paw edema model was employed. Each rat was
6
1
1
28.85, 129.80, 125.08, 124.78(2C), 123.86(2C), 114.08,
12.49, 96.24, 80.38, 56.59, 56.31, 56.06, 28.25(3C) ppm;
IR (KBr): νꢀ = 3045.2, 2980.5, 1760.5, 16.95.5,
−1
+
1150.9 cm ; MS (ESI+): m/z = 403.1 ([M + H] ), 425.1; anal
3
calcd for: C H N O : C, 62.67; H, 6.51; N, 6.96; O, 23.85;
injected with a freshly prepared 0.1 cm of 1% carrageen a
2
1 26 2 6
found: C, 62.65; H, 6.50; N, 6.98; O, 23.89.
suspension in normal saline (0.9% NaCl) into subplantar tis-
sue of the right hind paw. The intraperitoneal administration of
control, test samples (synthesized compounds) and reference
drug, for the control, 10 mg/kg saline solution was adminis-
tered. Paw edema was measured every 60 min for 3 h after
induction of inflammation. The anti-inflammatory activity of
the tested compounds and reference drug (ibuprofen 10 mg/
kg) were determined as the increase in paw edema volume
(control) and the results are summarized in Table 1 and as
percentage inhibition (% inhibition) and summarized in
Table 1. Results were expressed as the mean ± SE difference
between disease control (control) ten compounds and one
standard drug-treated animal using one-way analysis of vari-
ance ANOVA, followed by Dennett’s test for multiple
comparisons.
tert-butyl 2-(4-(1H-indol-2-yl)butanamido)phenylcarbamate
(
4i)
From the 0.150 g 4-(1H-indol-2-yl)butanoic acid (0.74 mmol,
3
1
eq), 0.140 cm DIPEA (1.11 mmol, 1.5 eq), 0.169 g com-
pound 3 (0.812 mmol, 1.1 eq), 0.212 g EDCI (1.11 mmol,
1
.5 eq) and 0.149 g HOBt (1.11 mmol, 1.5 eq) the compound
4
i was obtained as white colored solid (0.250 g, 75.7%) after
chromatography on a silica gel column with petroleum ether/
ethyl acetate (70:30, v/v). m.p.: 262–264 °C; H NMR
1
(
400 MHz, CDCl ): δ = 12.35 (s, 1H, NH), 9.42(s, 1H, NH),
3
8
.67 (s, 1H, NH), 7.98 (m, 4H, Ar-H), 7.63–7.42 (m, 2H, Ar-
H), 7.19 (d, 2H, J = 8.85 Hz, Ar-H), 6.72 (s, 1H, indole-H),
2
3
1
1
.62–2.41 (m, 4H, 2-CH -), 1.68 (q, 2H, −CH -), 1.45 (s, 9H,
2
2
13
CH ) ppm; C NMR (100 MHz, DMSO-d ): δ = 172.44,
Acute toxicity All animals used in the inflammatory experi-
ments were observed for 24 h and mortality of animals record-
ed where present for each group at the end of observation
3
6
54.18, 136.30, 130.82, 129.88, 127.33(2C), 126.19,
25.29(2C), 124.56, 121.74, 119.05, 118.71, 115.03, 111.15,

J Chem Biol
Table 1 The anti-inflammatory activity of the tert-butyl 2-(substituted benzamido) phenyl carbamate (4a–4j) derivatives and reference drug
(ibuprofen) in carrageenan-induced rat paw edema assay, values are expressed as mean ± SEM (mm) and percentage inhibition of inflammation
Compound codes Mean paw volume ± SEM and (% of inhibition)
1
h
3 h
6 h
9 h
12 h
4
4
4
4
4
4
4
4
4
4
a
b
c
d
e
f
0.823 ± 0.000
26.714)
0.783 ± 0.015
30.276)
0.631 ± 0.012
43.811)
0.733 ± 0.015
34.728)
0.782 ± 0.017
30.365)
0.676 ± 0.008
39.804)
0.634 ± 0.012
43.544)
0.702 ± 0.008
37.488)
0.822 ± 0.010
26.803)
0.893 ± 0.008
20.480)
0.851 ± 0.006
24.220)
1.123 ± 0.008
0.752 ± 0.000
(41.433)
0.524 ± 0.017
(54.553)
0.626 ± 0.010
(40.550)
0.422 ± 0.014
(54.130)***
(
0.651 ± 0.010
(49.299)
0.606 ± 0.010
(47.441)
0.652 ± 0.011
(38.081)
0.551 ± 0.012
(40.108)
(
0.593 ± 0.017
(53.816)
0.513 ± 0.008
(55.507)
0.417 ± 0.011
(60.398)
0.492 ± 0.025
(46.521)
(
0.510 ± 0.015
(60.280)
0.632 ± 0.015
(45.186)
0.582 ± 0.014
(44.729)
0.427 ± 0.008
(53.586)**
(
0.403 ± 0.012
(48.364)
0.625 ± 0.011
(45.793)
0.752 ± 0.011
(28.584)
0.561 ± 0.013
(39.021)
(
0.681 ± 0.015
(39.018)
0.651 ± 0.006
(43.538)
0.553 ± 0.020
(47.483)
0.425 ± 0.006
(53.804)**
(
g
h
i
0.653 ± 0.000
(49.143)
0.654 ± 0.020
(43.278)
0.602 ± 0.010
(42.830)
0.492 ± 0.014
(46.521)
(
0.652 ± 0.005
(49.221)
0.513 ± 0.011
(55.507)
0.535 ± 0.012
(49.192)
0.429 ± 0.012
(53.369)*
(
0.725 ± 0.006
(43.535)
0.682 ± 0.014
(40.849)
0.675 ± 0.014
(35.897)
0.421 ± 0.020
(54.239)***
(
j
0.642 ± 0.073
(50.000)
0.625 ± 0.010
(45.793)
0.622 ± 0.017
(40.930)
0.427 ± 0.011
(53.586)**
(
Indomethacin
0.752 ± 0.011
(41.433)
0.681 ± 0.008
(40.936)
0.527 ± 0.011
(49.952)
0.421 ± 0.012
(54.239)***
(
Disease control
Normal control
1.284 ± 0.006
1.153 ± 0.017
1.053 ± 0.088
0.920 ± 0.011
*
*The mean difference is significant at the P < 0.05 level; ***the mean difference is significant at the P < 0.01 level; Dunnetts-tests treat one group as a
control, and compare all other groups against it
period. Thus, edema volume in control (Vc) and in groups
treated with test compounds (Vt) was calculated. The percent-
age of inhibition was calculated using the formula
that the test drug treatment was replaced with appropriate
volumes of the vehicle. Indomethacin (10 mg/kg) for anti-
inflammatory was used as a reference drug.
Vc − Vt
Percentage of inhibition ¼
 100
Experimental animals Male Swiss albino rats (180–200 g)
were used for anti-inflammatory activity. All of the animals
were left for 2 days in the laboratory for acclimatization before
the day of experiment. A minimum of 6 animals were used in
each group. All pharmacological activities were carried out as
per CPCSEA (Committee for the Purpose of Control and
Supervision of Experiments on Animals) norms (Registered
No:1757/PO/RcBiBt/S/14 CPCSEA), after obtaining the ap-
proval from the Animal Ethics Committee from Jeeva life
science, Uppal Industrial area, Hyderabad, Telangana state,
India.
Vc
where Vc is mean paw inflammation of control animal and Vt
mean paw inflammation of test in drug-administered animal.
Statistical analysis The anti-inflammatory activity was deter-
mined as increase in the paw edema volume percentage in the
treated animals (Table 1). Results were expressed as the
mean ± SE, and different groups were compared using one-
way analysis of variance (ANOVA) followed by Dennett’s test
for multiple comparisons. Test where p < 0.05 was accepted to
be a significant difference.
Molecular docking
Preparation of test samples for bioassay All test samples
(
0
50 mg/kg) were suspended in a mixture of distilled water and
.5% carboxyl methylcellulose (CMC) and were given intra-
Crystal structure of cyclooxygenase-2 with PDB ID: 4COX
[9] was retrieved from the protein data bank. GLIDE 5.6 [28]
was used for molecular docking. Protein was prepared using
protein preparation wizard in Maestro 9.0 applying default
peritoneally (i.p.) to the test animals. The animals of the con-
trol group received the same experimental handling except

J Chem Biol
parameters. A grid was generated around the active site by
selecting the co-crystallized ligand. Receptor van der Waals
scaling for nonpolar atoms was kept at 0.9 [29] Molecules
were built using Maestro build panel and prepared by
LigPrep 2.0 application. Low-energy conformation of the li-
gands were selected and docked into the grid generated for the
protein using extra precision (XP) docking mode [30–34].
the compound displayed an intense molecular ion peak at m/z
+
239 [M] corresponding to molecular formula C H N O .
11 14 2 4
The resultant compound tert-butyl(2-nitrophenyl)carbamate
(2) further undergoes reduction with hydrazine hydrate (NH2-
NH .H O) in methanol to obtained the compound tert-butyl(2-
2
2
aminophenyl)carbamate (3) in excellent yield. The infrared
spectrum of compound 3 showed characteristic absorptions cor-
−
1
responding to amine (−NH ) is 3367 cm and it was also con-
2
1
firmed by the H NMR which showed broad singlet at 6.28 ppm
corresponding to amine protons attached to benzene ring and
singlet signal assigned at 9.15 ppm corresponding to NH proton
of adjusted phenyl ring. All the other aromatic protons of com-
pound 3 were observed at the expected regions. MS showed a
Results and discussion
Chemistry
+
molecular ion peak at m/z 209.0 [M] , corresponding to a mo-
The synthesis of the tert-butyl 2-(substituted benzamido)
phenylcarbamate derivatives (4a–4j) were illustrated in
Scheme 1. The compound 2-nitroaniline (1) was allowed to
undergo amine protection via nucleophilic addition reaction
by the tertiary butoxy carbonic anhydride (Boc) instanta-
neously in the presence of triethylamine to produced tert-butyl
lecular formula C H N O .
11 16 2 4
Finally, the compound tert-butyl(2-aminophenyl)carbamate
(3) was underwent amidation coupling with different carboxylic
acids, and combination of 1-(3-dimethyl amino propyl)-3-
ethylcarbodiimide.HCl (EDCI) as the carboxylate activator
and hydroxyl benzotriazole (HOBt) at room temperature in pres-
ence DIPEA as base to obtain the corresponding tert-butyl
2-(substituted benzamido)phenylcarbamate (4a–4j) in moderate
to be good yields [36] indicated in Scheme 1. Moreover, amide
synthesis using coupling reagent EDCI is low recrimination
when used with HOBt reagent by product both easily soluble
in water. The structures of all the newly synthesized compounds
(
[
2-nitrophenyl) carbamate (2) in moderate to be good yield
35]. The structure of the compound was established on the
1
basis of its spectral properties (IR, H NMR, MS, and elemen-
tal analysis). The IR spectrum showed the presence of absorp-
tion bands at 3360 and 1680 cm due to NH and C = Os
−1
1
functional group, respectively. H NMR spectrum showed
1
13
broad singlet signals at 9.19 ppm, assigned to NH groups;
the signal of up-field singlet was assigned to tertiary butoxy-
proton at 1.45 ppm, in addition to the multiple signals of
aromatic protons in the region of 8.53–7.78 ppm. The MS of
4a–4j were confirmed by IR, H NMR, C NMR, and MS and
elemental analysis spectra data are included experimental sec-
tion. In the IR spectra of the compound 4a amide NH- and Boc-
protected NH group absorption peak appeared at 3266 and
Scheme 1 Synthesis of tert-butyl
(substituted
benzamido)phenylcarbamate 4a–
j

J Chem Biol
Fig. 1 Graphical representation
percentage of inhibition of
compounds 4a–4j along with the
standard drug
1
3
1
034 cm− respectively. The absorption peak appeared at
two carbonyl groups appeared at 165.9 and 164.6 ppm. Boc
linkage carbon exhibited resonance peaks at 81.2, 28.19 ppm.
The carbon atoms of aromatic ring appeared at 154.6–
115.4 ppm. MS showed a molecular ion peak at m/z 331.05
693 cm− was attributed to C = O group of phenylcarbamate.
1
In addition, the absorption bands, corresponding to C = O of the
amide moiety, were observed at 1650 cm . The vibration fre-
quency of C–H and C = C group possible to identify substitution
pattern on benzene appeared at 2980 and 1693 cm , respec-
tively. In addition, substituted mono halo on benzene ring ab-
sorption strong peak appeared at 769 cm . Thus, the data of IR
spectroscopy supported the formation of the desired structure.
The H NMR spectrum showed signals of two singlets which
−1
+
[M] , corresponding to a molecular formula C H N O . All
23 38 2 3
−1
the synthesized compounds exhibited satisfactory spectral data
consistent with their molecular structures as shown in Scheme 1.
−1
Anti-inflammatory activity
1
were assigned to amide and phenyl carbamate at 9.48 and
The in vivo anti-inflammatory activity [37–39] evaluation of
title compounds (4a–4j) were carried out using carrageenan-
induced paw edema assay of Winter et al. and the result are
summarized in Table 1. The Male Swiss albino rats at
10 mg/kg body weight and the standard drug, indomethacin
9
1
.05 ppm, respectively. A signal of Boc protons appears at
.48 ppm in addition to the signal of downfield singlet of aro-
matic protons in the region of 8.00–6.80 ppm. The structure was
13
further confirmed by C NMR signals of carbon atoms of the
Table 2 Docking scores and ADME properties of synthesized compounds
a
b
c
d
e
f
Title
Dock score
M.Wt
QPlogPo/w
QPlogS
QPPCaco
QPlogBB
QPPMDCK
% Human oral absorption
4
4
4
4
4
4
4
4
4
4
a
b
c
d
e
f
−8.49
−5.33
−7.93
−5.98
−5.17
−6.60
−8.08
−5.62
−9.56
−6.20
−11.1
330.36
326.39
368.47
438.26
402.36
390.56
382.79
402.45
393.48
329.19
357.79
3.479
3.566
4.457
3.786
2.038
5.636
4.109
3.673
4.588
2.502
4.261
−4.568
−4.626
−5.293
−4.68
−4.125
−6.428
−5.322
−4.513
−5.565
−3.323
−5.119
1470.54
1538.17
1616.79
1709.55
39.209
−0.338
−0.448
−0.486
−0.271
−2.087
−1.135
−0.163
−0.544
−0.891
−0.29
1358.19
787.91
831.53
1676.76
14.92
100
100
100
100
54.44
1552.48
1490.37
1971.05
965.68
795.84
3798.53
1030.11
476.38
1414.68
258.70
100
100
100
100
100
g
h
i
j
1426.09
190.46
IMC
−0.609
92.697
IMC indomethacin
a
Predicted octanol/water partition coefficient log P (acceptable range—2.0–6.5)
Predicted aqueous solubility in mol/L (acceptable range—6.5–0.5)
b
c
d
e
f
Predicted caco cell permeability in nm/s (acceptable range: <25 is poor and >500 is great)
Predicted blood brain barrier permeability (acceptable range—3–1.2)
Predicted apparent MDCK cell permeability in nm/s (acceptable range: <25 is poor and >500 is great)
Percentage of human oral absorption (acceptable range: <25 is poor and >80% is high

J Chem Biol
used for anti-inflammatory activity, all the exhibited deriva-
tives of anti-inflammatory activities that listed for every 3 h
with potency that increased with time. The results were
expressed as the increase in paw volume at various time inter-
vals in comparison to the initial values shower in Table 1. The
increase of volumes in percentage was calculated by
subtracting the initial paw volumes from the paw volumes
obtained after the carrageenan agent was injected.
result in comparison with the standard drug, indomethacin,
may be due to amide moiety enhancing the activity.
Among them the compounds 4a, 4d, 4 h, 4i, and 4j showed
maximum activity and remaining compounds 4b, 4c, and 3g
showed moderate to good activity after 12 h observation. In
the present investigation, the highest activity of compound 4a
and 4i than standard may be attributed to the presence of
highly electronegative florosubstituted on phenyl ring and in-
dole moiety, respectively, have been playing vital role for
activity. It is found that the compounds with viz. electron
The inflammatory activity was tested for ten desired com-
pounds. Most of the tested compounds have shown an better
Fig. 2 Dock poses conformation
of active compounds 4a (a), 4i
(b), and standard drug
indomethacin (c) in COX-2
protein active site

J Chem Biol
withdrawing nitro group 4e (39.021%), showed less activity.
All the synthesized compounds with their anti-inflammatory
activity and percentage of inhibition are presented in Table 1.
It was found that better anti-inflammatory activity for 4a
of indomethacin showed in Fig. 2c, the dock score value of
−11.14 kcal/mol(bond length of 1.989 A ). Bonding interac-
tions and bond lengths of active compounds with COX-2
protein was tabulated in Table 3. Finally compounds 4a and
4i were found to be potent and have high docking score with
the COX-2 binding site.
o
(
54.130%) and 4i (54.239%) compared to standard drug after
2-h intervals was observed in the present experiment (Fig. 1).
1
The degree of anti-inflammatory action in ascending order in
percentageofinhibitionis39.021(4e)<40.108(4b)<46.521(4c)-
ADME (absorption, distribution, metabolism, and excre-
tion) properties of synthesized compounds were evaluated
computationally using QikProp module of Schrodinger
(QikProp3.4, Schrödinger,LLC, New York, NY, 2010) and
were analyzed for drug-likeness by applying Lipinski’s rule
of five (Table 2) are calculated by QikProp. All these pharma-
cokinetic parameters were found to be good and acceptable
range.
In addition a regression analysis between dock score (bind-
ing affinity) and degree of anti-inflammatory activity values of
the synthesized molecules were carried out. It gave a correla-
tion coefficient r of 0.828 representing significant correlation
between binding affinity (dock score) and degree of anti-
inflammatory activity.
=46.521(4g)<53.369(4h)<53.586(4j)=53.586(4d)<53.804(4f)-
<
54.130(4a) < 54.239(4i), and 54.239 standard drug, indo-
methacin, by the percentage of inhibition of carrageenan-
induced paw edema showed in Table 1. In all the experiments
after the 12-h observation, it showed a normal stage in the entire
paw but it was observed (disease control) carrageenan paw with
some inflammatory and also showed normal paw. The feet of
rat paw edema injection of carrageen injection after 3 h, it was
considered 0 h showed in Table 1.
In silico molecular docking
To gain more insight into the interactions of the high active
compounds in the series, molecular docking of 4a–4j was
performed into the active site of COX-2. The molecules were
deeply embedded into the hydrophobic pocket in the active
site similar to indomethacin (standard). The compounds
showed hydrogen bond interaction with Ser 530, Try 355,
and Arg 120, π-π interactions with Trp 387, Tyr 385 amino
acids, and the range of dock score from −9.56 to −5.17 kcal/
mol. The docking score of all the compounds are depicted in
Table 2. The best active compound 4a, showed two hydrogen
Conclusions
In conclusion, a new class of tert-butyl 2-(substituted
benzamido) phenylcarbamate (4a–4j) has been synthesized
from tert-butyl 2-amino phenylcarbamate (3) and character-
ized by spectroscopic techniques. All the synthesized com-
pounds were evaluated for their anti-inflammatory. Among
the entire compound, the compounds 4a, 4d, 4f, 3i and 3j
were showed maximum activity within 9 to 12 h. Moreover,
molecular docking study was performed to provide the bind-
ing patterns of the compound 4a and 4i into the binding sites
of COX-2 (PDB code: 4COX) enzymes. The study showed
that 4a and 4i has favorable orientation within the COX-2
enzyme binding site and has a high docking score. In view
of these studies, these compounds could be a subject of further
investigations for searching potential new anti-inflammatory
molecules.
bond interactions with Try 355 and Arg 120 (bond length of
o
2
.389 and 1.720 A ) with a dock score of −8.49 kcal/mol.
Additionally, it showed two π-π interactions with Trp 387
and Tyr 385 when compared to standard drug indomethacin,
this can be explained in terms of hydrophobic interaction with
the COX-2 protein active site (Fig. 2a). Another compound 4i
showed one hydrogen bond interaction with Ser 530, like
Indomethacin, with the dock score value of −9.56 kcal/mol
o
(
bond length of 2.069 A ) (Fig. 2b). Dock pose conformation
Table 3 Bonding interactions
and bond lengths of active
compounds with COX-2 protein
o)
Title comp.
Bonding interactions
Protein amino acids
Tyr 355 and Arg 120
Bond length (A
4
a
Two hydrogen bonding interactions
2.389 and 1.720
Two hydrophobic π-π interactions
One hydrogen bond interaction
Trp 387 and Tyr 385
Arg 120
4
4
4
4
4
c
f
2.162
2.083
1.952
1.974
2.069
1.989
One hydrogen bond interaction
One hydrogen bond interaction
One hydrogen bond interaction
One hydrogen bond interaction
One hydrogen bond interaction
One hydrophobic π-π interaction
Arg 120
Ser 530
Ser 530
Ser 530
Ser 530
Tyr 385
g
h
i
IMC

J Chem Biol
Acknowledgements The author, Bhookya Shankar thanks to UGC-
BSR, (RFSMS-Award No. 805/chem/2013), NEW Delhi, India for finan-
cial support in the form of senior research fellowship (SRF) and we thank
Indian institute of chemical technology (IICT), Hyderabad, for providing
the facilities of spectral analysis. We also thank to Jeeva life science, IDA
Uppal, Hyderabad, Telangana, India for providing facilities of biological
activity.
15. Bielawska A, Bielawski K, Muszynska A (2004) Synthesis and
biological evaluation of new cyclic amidine analogs of
chlorambucil. Il Farmaco 59:111–117
16. Ozden S, Atabey D, Yildiz S, Goker H (2005) Synthesis and potent
antimicrobial activity of some novel methyl or ethyl 1H-
benzimidazole-5-carboxylates derivatives carrying amide or
amidine groups. Bioorg Med Chem 13:1587–1597
1
7. Zhuravel IO, Kovalenko SM, Ivachtchenko AV, Balakin KV,
Kazmirchuk VV (2005) Synthesis and antimicrobial activity of 5-
hydroxymethyl- 8-methyl-2-(N-arylimino)-pyrano[2,3-c]pyridine-
References
3
-(N-aryl)-carboxamides. Bioorg Med Chem Lett 15:5483–5484
1
8. Rudolph MJ, Illig CR, Subasinghe NL, Wilson KJ, Hoffman JB,
Randle T, Green D, Molloy CT, Maguire D, Grasberger BL,
Desjarlais RL, Zhou Z (2002) Design and synthesis of 4,5-disub-
stituted-thiophene-2-amidines as potent urokinase inhibitors.
Bioorg Med Chem Lett 12:491–495
1
.
.
Sondhi SM, Singhal N, Johar M, Reddy BSN, Lown JW (2002)
Heterocyclic compounds as inflammation inhibitors. Curr Med
Chem 9:1045–1074
Chan CC, Boyce S, Brideatt C, Ford-Hutchinson AW, Gordon R,
Guay D, Hill RG, Li CS, Mancini J, Penneton M (1995)
Pharmacology of a selective cyclooxygenase-2 inhibitor, L-745,
2
1
2
9. Griffin MR, Yared A, Ray WA (2000) Nonsteroidal
antiinflammatory drugs and acute renal failure in elderly persons.
Am J Epidemiol 151:488–496
3
37: a novel nonsteroidal anti-inflammatory agent with an ulcero-
0. Ukrainets IV, Sidorenko LV, Petrushovo LA, Gorokhova OV
genic sparing effect in rat and nonhuman primate stomach. J
Pharmacol Exp Ther 274(3):1531–1537
Bentolini A, Ottani A, Sandrini M (2002) Selective COX-2 inhib-
itors and dual acting anti-inflammatory drugs: critical remarks. Curr
Med Chem 9:1033–1043
(2006) Synthesis of amidine and amide derivatives and their eval-
uation for anti-inflammatory and analgesic activities. Chem
Heterocycl Comput 42:64
3
.
.
2
1. Chan FK (2006) Primer: managing NSAID-induced ulcer compli-
cations balancing gastrointestinal and cardiovascular risks. Nat Clin
Pract Gastroenterol Hepatol 3:563–573
4
Schneider V, Lévesque LE, Zhang B, Hutchinson T, Brophy JM
(
2006) Association of selective and conventional nonsteroidal
2
2
2
2. Dannhardt G, Kiefer W (2001) Cyclooxygenase inhibitors—cur-
rent status and future prospects. Eur J Med Chem 36:109–126
3. Burnier M (2005) The safety of rofecoxib. Expert Opin Drug Saf 4:
antiinflammatory drugs with acute renal failure: a population-based,
nested case-control analysis. Am J Epidemiol 164:881–889
Simon LS (2001) COX-2 inhibition: an advance or only pharma-
ceutical "hype"? Arthritis Care Res 45:209–215
Kalgutkar AS, Crews BC, Rowlinson SW, Marnett AB, Kozak KR,
Remmel KR, Marnett LJ (2000) Biochemically based design of
cyclooxygenase-2 (COX-2) inhibitors: facile conversion of nonste-
roidal antiinflammatory drugs to potent and highly selective COX-2
inhibitors. Proc Natl Acad Sci U S A 97:925–930
5
6
.
.
4
91–499
4. Galanakis D, Kourounakis AP, Tsiakitzis KC, Doulgkeris C, Rekka
EA, Gavalas A, Kravaritou C, Christos C, Kourounakis PN (2004)
Synthesis and pharmacological evaluation of amide conjugates of
NSAIDs with L-cysteine ethyl ester, combining potent
antiinflammatory and antioxidant properties with significantly re-
duced gastrointestinal toxicity. Bioorg Med Chem Lett 14:3639–
7
.
Kalgutkar AS, Marnett AB, Crews BC, Remmel RP, Marnett LJ
3
643
(
2000) Ester and amide derivatives of the nonsteroidal
2
5. Kumar P, Knaus EE (1993) Synthesis and antiinflammatory activity
of 5-(1,2-dihydropyridyl)-tetrazol-2-acetic acids, esters and amides.
Eur J Med Chem 28:881–885
antiinflammatory drug, indomethacin, as selective
cyclooxygenase-2 inhibitors. J Med Chem 43:2860–2870
Haggar RE, Al-Wabli RI (2015) Anti-inflammatory screening and
molecular modeling of some novel coumarin derivatives.
Molecules 20:5374–5391
Kurumbail GR, Stevens MA, Gierse KJ, Mc Donald JJ, Stegeman
AR, Pak YJ, Gildehaus D, Miyashiro MJ, Pennung DT (1996)
Structural basis for selective inhibition of cyclooxygenase-2 by
anti-inflammatory agents. Nature 384:644–648
8
.
.
2
6. Ban M, Taguchi H, Katushima T, Takahashi M, Shinoda K,
Watanabe A, Tominaga T (1998) Novel antiallergic and
antiinflammatory agents. Part I: synthesis and pharmacology of
glycolic amide derivatives. Bioorg Med Chem 6:1069–1076
7. Sondhi SM, Sing J, Kumar A, Jamal H, Gupta PP (2009) Synthesis
of amidine and amide derivatives and their evaluation for anti-
inflammatory and analgesic activities. Eur J Med Chem 44:1010–
9
2
1
0. Scheiman JM (2005) Effects of nonsteroidal anti-inflammatory
drugs, including COX-2 specific inhibitors, on the GI tract.
ASGE Clinical Update 12:1–4
1
015
2
2
8. Glide Version 5.6. Schrödinger, LLC: New York, 2010
9. Friesner RA, Banks JL, Murphy RB (2004) Glide: a new approach
for rapid, accurate docking and scoring. 1. Method and assessment
of docking accuracy. J Med Chem 47:1739–1749
1
1. Glushkov VA, Anikina LV, Vikharev YB, Feshina EV, Shklyaev
YV (2005) Synthesis and antiinflammatory and analgesic activity
of amidines of 3,4-dihydroisoquinoline series. Pharm Chem J 39:
3
0. Bharate SB, Nagaraju M, Sudhakar M, Bharate JB, Ramesh M,
Yadav RM, Vishwakarma RA (2012) Efficient synthesis of 2,3-
dihydroquinazolin-4(1H)-ones using heterogeneous solid acid cat-
alysts: unexpected formation of 2,3-dihydro-2-(4-(tetrahydro-2H-
pyran-2-yloxy)butyl)quinazolin-4(1H)-one. ARKIVOC 8:308–318
5
33–536
2. Panico A, Vinici P, Inert M, Cardile V, Gentile B, Ronsisvalle G
2002) Amidinobenzisothiazole derivatives with antidegenerative
1
1
(
activity on cartilage. Farmaco 57:671–675
3. Sielecki TM, Liu J, Mousa SA, Racanelli AL, Hausner EA, Wexler
RR, Olson RE (2001) Synthesis and pharmacology of modified
amidine isoxazoline glycoprotein IIb/IIIa receptor antagonists.
Bioorg Med Chem Lett 11:2201–2204
4. Sienkiewicz P, Bielawski K, Bielawska A, Palka J (2005) Inhibition
of collagen and DNA biosynthesis by a novel amidine analogue of
chlorambucil is accompanied by deregulation of β(1)-integrin and
IGF-I receptor signaling in MDA-MB 231 cells. Environ Toxicol
pharmacol 20:118–24
31. Omar Al-Deeb A (2008) Synthesis of some new 3H-quinazolin-4-
one derivatives as potential antitubercular agents. World Applied
Sciences Journal 5:94–99
32. Shankar B, Jalapathi P, Ramesh M, Kishore Kumar A, Ragavender
M, Bharath G (2016) Synthesis, antimicrobial evaluation, and
docking studies of some novel benzofuran based analogues of
chalcone and 1,4-benzodiazepine. Russ J Gen Chem 86:1711–1721
33. Kishore Kumar A, Sunitha V, Shankar B, Ramesh M, Murali
Krishna T, Jalapathi P (2016) Synthesis, biological evaluation,
1

J Chem Biol
and molecular docking studies of novel 1,2,3-triazole derivatives as
37. Winter CA, Risley EA, Nuss GW (1962) Carrageenin-induced ede-
ma in hind paw of the rat as an assay for antiinflammatory drugs.
Expe Bio Mad 111:544–547
potent anti-inflammatory agents. Russ J Gen Chem 86:1154–1162
3
4. Shankar B, Jalapathi P, Nagamani M, Bharath G, Karunakar Rao K
(
2016) Monatsh Chem. doi:10.1007/s00706-016-1838-z
38. Shankar B, Jalapathi P, Sunitha V, Karunakar rao K (2016)
Synthesis of 5, 5′-methylenebis ((benzofuran-5,2-
diyl))bis(phenylmethanone) and its phenylhydrazonoe,
benzenesulfonate derivatives and their anti-inflammatory activity.
Der Pharma Chemica 8:192–199
39. Amir M, Akhter MW, Alam O (2016) Synthesis, characterization,
and biological evaluation of furoxan coupled ibuprofen derivatives
as anti-inflammatory agents. Monatsh Chem 147:493–508
3
5. Jalapathi P, Anil V, Ramesh M, Bhavani AKD, Umapathi N,
Parthasarathy T, Bharath G, Gangagnirao A (2014) Synthesis and
molecular modeling studies of novel tert-butyl 2, 4-disubstituted
carboxamido phenylcarbamate derivatives and evaluation of their
antimicrobial activity. Der Pharma Chemica 6:61–76
3
6. Carpino LA (1993) 1-Hydroxy-7-azabenzotriazole. An efficient
peptide coupling additive J Am Chem Soc 115:4397–4398