Synthesis, anti-inflammatory, cytotoxic, and COX-1/2 inhibitory activities of cyclic imides bearing 3-benzenesulfonamide, oxime, and β-phenylalanine scaffolds: a molecular docking study

Article abstract of DOI:10.1080/14756366.2020.1722120

Cyclic imides containing 3-benzenesulfonamide, oxime, and β-phenylalanine derivatives were synthesised and evaluated to elucidate their in?vivo anti-inflammatory and ulcerogenic activity and in?vitro cytotoxic effects. Most active anti-inflammatory agents

Full text of DOI:10.1080/14756366.2020.1722120

Journal of Enzyme Inhibition and Medicinal Chemistry
ISSN: 1475-6366 (Print) 1475-6374 (Online) Journal homepage: https://www.tandfonline.com/loi/ienz20
Synthesis, anti-inflammatory, cytotoxic, and
COX-1/2 inhibitory activities of cyclic imides
bearing 3-benzenesulfonamide, oxime, and β-
phenylalanine scaffolds: a molecular docking
study
Alaa A.-M. Abdel-Aziz, Adel S. El-Azab, Nawaf A. AlSaif, Mohammed M.
Alanazi, Manal A. El-Gendy, Ahmad J. Obaidullah, Hamad M. Alkahtani,
Abdulrahman A. Almehizia & Ibrahim A. Al-Suwaidan
To cite this article: Alaa A.-M. Abdel-Aziz, Adel S. El-Azab, Nawaf A. AlSaif, Mohammed M.
Alanazi, Manal A. El-Gendy, Ahmad J. Obaidullah, Hamad M. Alkahtani, Abdulrahman A. Almehizia
& Ibrahim A. Al-Suwaidan (2020) Synthesis, anti-inflammatory, cytotoxic, and COX-1/2 inhibitory
activities of cyclic imides bearing 3-benzenesulfonamide, oxime, and β-phenylalanine scaffolds:
a molecular docking study, Journal of Enzyme Inhibition and Medicinal Chemistry, 35:1, 610-621,
DOI: 10.1080/14756366.2020.1722120
To link to this article: https://doi.org/10.1080/14756366.2020.1722120
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JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
2020, VOL. 35, NO. 1, 610–621
https://doi.org/10.1080/14756366.2020.1722120
RESEARCH PAPER
Synthesis, anti-inflammatory, cytotoxic, and COX-1/2 inhibitory activities of cyclic
imides bearing 3-benzenesulfonamide, oxime, and b-phenylalanine scaffolds:
a molecular docking study
Alaa A.-M. Abdel-Aziz , Adel S. El-Azab, Nawaf A. AlSaif, Mohammed M. Alanazi, Manal A. El-Gendy,
Ahmad J. Obaidullah, Hamad M. Alkahtani, Abdulrahman A. Almehizia
and Ibrahim A. Al-Suwaidan
Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia
ABSTRACT
ARTICLE HISTORY
Received 21 December 2019
Revised 14 January 2020
Accepted 19 January 2020
Cyclic imides containing 3-benzenesulfonamide, oxime, and b-phenylalanine derivatives were synthesised
and evaluated to elucidate their in vivo anti-inflammatory and ulcerogenic activity and in vitro cytotoxic
effects. Most active anti-inflammatory agents were subjected to in vitro COX-1/2 inhibition assay. 3-
Benzenesulfonamides (2–4, and 9), oximes (11–13), and b-phenylalanine derivative (18) showed potential
anti-inflammatory activities with 71.2–82.9% oedema inhibition relative to celecoxib and diclofenac (85.6
and 83.4%, respectively). Most active cyclic imides 4, 9, 12, 13, and 18 possessed ED50 of
35.4–45.3 mg kgꢀ1 relative to that of celecoxib (34.1 mg kgꢀ1). For the cytotoxic evaluation, the selected
derivatives 2–6 and 8 exhibited weak positive cytotoxic effects (PCE ¼ 2/59–5/59) at 10 lM compared to
the standard drug, imatinib (PCE ¼ 20/59). Cyclic imides bearing 3-benzenesulfonamide (2–5, and 9), ace-
tophenone oxime (11–14, 18, and 19) exhibited high selectivity against COX-2 with SI > 55.6–333.3 rela-
tive to that for celecoxib [SI > 387.6]. b-Phenylalanine derivatives 21–24 and 28 were non-selective
towards COX-1/2 isozymes as indicated by their SI of 0.46–0.68.
KEYWORDS
Cyclic imide; anti-
inflammatory activity;
cytotoxic activity; COX-1/2
inhibition; molecu-
lar docking
1. Introduction
bearing a-amino acids are reported as promising COX-1/COX-2
inhibitors (Figure 1(G,H))15.
Nonsteroidal anti-inflammatory drugs (NSAIDs) are the drugs of
choice for the treatment of inflammation and pain1,2. NSAIDs are
cyclooxygenase inhibitors and they include the COX-1 and COX-2
enzymes1–3. The cyclooxygenase isozymes are responsible for the
Here, we report the design and synthesis of novel cyclic imides
containing 3-benzenesulfonamide, acetophenone oxime, and
b-phenylalanine scaffolds. The designed cyclic imides (Figure 1(I))
were subjected to various analyses to: (i) investigate their in vivo
anti-inflammatory and ulcerogenic activities, and their in vitro
COX-1/2 inhibitory effects; (ii) study their in vitro cytotoxicity activ-
ity; (iii) explore their structure-activity relationships (SAR) based on
their in vivo anti-inflammatory effects and in vitro COX-1/2 inhibi-
tory activity with molecules containing substituted cyclic imides;
(iv) compare the biological effects of 3-benzenesulfonamide to
those of acetophenone oxime and b-phenylalanine based on
in vivo anti-inflammatory and COX-1/2 inhibition; and (v) conduct
a molecular docking study of the target derivatives to investigate
their binding with the COX-2 isozyme.
conversion of arachidonic acid to eicosanoids (prostaglandins)1–3
.
In this process, the inhibition of the COX-2 isozyme elucidates the
anti-inflammatory effects of NSAIDs1–3 while the inhibition of the
COX-1 isozyme is responsible for the major side effects displayed
by NSAIDs, such as gastrointestinal implication1,2. Alternatively,
selective COX-2 inhibitors, including compounds that contain
the pyrazole ring system, such as celecoxib (A) and SC-558 (B)4
(Figure 1), display anti-inflammatory activity with improved gastric
profile protection compared to NSAIDs4. Moreover, celecoxib, a
COX-2 inhibitor (Figure 1(A)) that displays an anti-inflammatory
effect, has been investigated as an antitumor agent5.
Compounds containing the sulphonamide moiety are interest-
ing derivatives that possess versatile and potential biological activ-
ities, such as carbonic anhydrase inhibitors6–9, COX-1/2 inhibition
(Figure 1(C–F))6a,10,11, and anti-inflammatory (Figure 1(C–F))6a,10,11
and antitumor activities6a,12. Recently, we reported the synthesis
2. Results and discussion
2.1. Chemistry
The chemistry for the synthesis of the target molecules 2–10,
11–19, and 21–29 is shown in Schemes 1–3. The rationale used
for the synthesis of these compounds is based on two routes
of phthalimide derivatives with potential anti-inflammatory6a,10,11
,
antitumor6a,13
, hypoglycaemic, and antihyperlipidemic proper-
ties14. In addition, some of these compounds were identified to where a non-carboxylic moiety, such as 3-benzenesulfonamide
display potent COX-2 and carbonic anhydrase inhibitory activ- and hydroxyiminoethyl (oxime), and a carboxylic moiety, such as
ities6–11
.
Recently, phthalimide analogues such isoindolines an amino acid (b-phenylalanine), are inserted into a versatile cyclic
CONTACT Alaa A.-M. Abdel-Aziz
almoenes@ksu.edu.sa
Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, P.O. Box 2457,
Riyadh 11451, Saudi Arabia
ß 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use,
distribution, and reproduction in any medium, provided the original work is properly cited.

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
611
H2NO2S
H2NO2S
O
N
N
N
N
CF3
CF3
SO2NH2
N
O
H3C
Br
SC-558 (B)
(C)
Celecoxib (A)
SO2NH2
O
O
N
O
O
HO
COOH
SO2NH2
N
N
O
O
O
(D)
(E)
(F)
N
N
COOH
COOH
(G)
(H)
Molecular design of small molecules as COX-1/COX-2 inhibitors
Structure modification
O
O
SO2NH2
O
N
O
NOH
N
N
COOH
O
O
O
O
SO2NH2
O
N
O
NOH
NOH
N
N
COOH
O
O
O
SO2NH2
O
N
O
O
N
N
COOH
O
O
Designed target molecules (I)
Figure 1. Some reported selective COX-2 inhibitors (A–H) and the designed compounds (I).
imides to explore and compare the efficacy of the substituted cyc- 3-isoindolinediones and hydroxylamine hydrochloride in glacial
lic imides for inhibiting COX-1/COX-2 and evaluating their anti- acetic acid containing anhydrous sodium acetate6b. The acid
inflammatory activity.
imides 21–29 were synthesised as indicated in Scheme 3. The
3-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoic acid derivatives
21–29 were obtained with 66–93% yield by refluxing the
cyclic imides and b-phenylalanines in glacial acetic acid contain-
Non-carboxylic cyclic imide-based 3-benzenesulfonamides
2–10 and oximes 11–19 were synthesised as indicated in
Schemes 1 and 2. Different carboxylic acid anhydrides were
refluxed with 3-aminobenzenesulfonamide (1) in glacial acetic ing anhydrous sodium acetate (Scheme 3). The molecular struc-
acid in the presence of anhydrous sodium acetate6–11 to obtain tures of the newly synthesised compounds 2–10, 11–19, and
cyclic imides 2–10 of good yields (Scheme 1). Scheme 2 displays 21–29 were verified by spectral analyses including mass, IR, 1H
the process used to obtain the 2-[4-(1-(hydroxyimino)ethyl)- NMR, and 13C NMR spectra as indicated in the experimen-
phenyl] derivatives 11–19 via the stirring of 2-(4-acetylphenyl)-1, tal section.

612
A. A.-M. ABDEL-AZIZ ET AL.
O
O
O
O
O
O
H2N
S
H2N
S
N
N
O
3O
2
O
O
O
H2N
S
N
O
O
O
R
H2N
S
O
O
Acetic acid, sodium acetate
Reflux, 12h
+
NH2
R= 4; H, 5; 5-Me, 6; 5-tert-But, 7; 5,6-Dichloro,
8; 4,5,6,7-Tetrachloro,9; 5-NO2,
O
1
Anhydrides
O
O
O
S
N
H2N
O
10
Scheme 1. Synthesis of 1,3-isoindolinediones incorporating 3-benzenesulfonamide 2–10.
OH
O
OH
O
N
N
N
N
O
12
O
11
OH
O
N
N
O
R
O
Acetic acid, sodium acetate
NH2OH, rt, 12h
N
R= 13; H, 14; 5-Me, 15; 5-tert-But, 16; 5,6-Dichloro,
17; 4,5,6,7-Tetrachloro, 18; 5-NO2
O
O
cyclic imides
O
N
OH
N
O
19
Scheme 2. Synthesis of 1,3-isoindolinediones incorporating oxime 11–19.
drugs, diclofenac and celecoxib, caused 83.4 and 85.6% oedema
inhibition, respectively. The non-carboxylic derivatives 2–10 (sul-
phonamide) and 11–19 (oxime) were generally more potent than
the corresponding carboxylic acid derivatives 21–29 (b-phenyl-
alanine), as observed for the non-carboxylic cyclic imides 2–4, 9,
11–13, and 18. These derivatives displayed the highest anti-
inflammatory activities with 71.2–82.9% paw oedema reduction
compared to the acidic derivatives 21–23 and 28, with
42.5–64.4% paw oedema reduction. For sulphonamide derivatives
2–10, the succinimide derivative 2 displayed a greater anti-inflam-
matory activity than the naphthalimide derivative 10, with per-
2.2. Biological activity
2.2.1. In vivo anti-inflammatory activity and SAR study
Twenty-seven compounds, 2–10, 11–19, and 21–29 and the refer-
ence drugs, diclofenac and celecoxib, were administered to the
rat carrageenan-induced foot paw oedema model to explore
the in vivo anti-inflammatory activities of such compounds. The
percentage inhibition of paw oedema was calculated after 2 h of
carrageenan treatment when maximal inhibition of carrageenan-
induced paw oedema was observed (Table 1)3,16,17
Data in Table 1 show that most of the designed compounds
.
decreased paw oedema by 21.2–82.9% (Table 1). The reference centage paw oedema reduction of 71.2 and 49.2%, respectively.

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
613
Scheme 3. Synthesis of 1,3-isoindolinediones incorporating b-phenylalanine 21–29.
Table 1. Results of anti-inflammatory activity of the tested compounds against carrageenan induced rat paw oedema in ratsa.
% Inhibition of paw
oedema from
control group
% Inhibition of paw
oedema from
control group
Mean %a increase in
paw weight SEMb
Mean %a increase in
paw weight SEMb
Compound No.
Compound No.
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
53.2 0.4
42.4 0.2
41.0 0.2
78.1 0.7
106.1 0.9
129.7 0.9
134.0 0.9
35.6 0.1
93.7 0.4
45.0 0.3
37.5 0.5
36.2 0.2
70.8 0.6
95.7 0.9
115.3 1.2
71.2 0.5
77.0 0.6
77.8 0.6
57.7 0.5
40.9 0.6
29.7 0.2
26.9 0.5
80.9 0.9
49.2 0.6
75.6 0.6
79.7 0.4
80.4 0.7
61.6 0.5
48.1 0.2
39.6 0.4
17
18
19
21
22
23
24
25
26
27
28
29
Celecoxib
diclofenac
135.6 0.8
31.6 0.4
57.6 0.5
98.9 0.6
69.1 0.5
65.6 0.3
123.4 0.9
129.0 1.1
136.3 0.8
141.6 1.0
100.9 0.8
130.8 0.7
27.3 0.4
30.1 0.3
25.1 0.6
82.9 0.8
68.8 0.4
46.4 0.3
62.9 0.4
64.4 0.7
33.1 0.4
29.6 0.7
24.4 0.3
21.2 0.9
42.5 0.2
27.0 0.1
85.6 0.4
83.4 0.5
aResult of control group (%): 165.6 1.6. bSignificant difference from control and celecoxib-treated group using unpaired Student’s t-test p < 0.05.
The conversion of succinimide 2 to the corresponding tetrahy- that the derivatives containing succinimide 11, tetrahydrophthali-
drophthalimide 3 or phthalimide 4 led to an increase in the anti- mide 12, phthalimide 13, and 5-nitro phthalimide 18 had anti-
inflammatory activity, with percentage paw oedema reduction of inflammatory activity levels (75.6, 79.7, 80.4, and 82.9% oedema
77.0 and 77.8%, respectively. Electronic substitution on the phthali- inhibition, respectively) higher than the corresponding analogues
mide core has a direct effect on the anti-inflammatory effects. For 14–17 and 19 (61.6, 48.1, 39.6, 25.1, and 68.8% oedema inhibition,
example, the presence of the methyl or tert-butyl group at the respectively). Replacing 3-benzenesulfonamide with compounds
5-position led to a decrease in activity, as observed for compounds that had a terminal carboxylic moiety at the cyclic imide core
5 and 6, which had a percentage paw oedema reduction of 57.7 21–29 caused a decrease in anti-inflammatory levels, with percent-
and 40.9%, respectively. Replacing the alkyl group with halogen age oedema inhibition ranging from 21.2–64.4%. Similarly, the
atoms resulted in derivatives with weak anti-inflammatory activity phthalimide derivative 23 had a higher activity than succinimide 21
as observed for compounds 7 and 8 (29.7 and 26.9% paw oedema or naphthalimide 29, with percentage paw oedema reduction of
reduction, respectively). However, modify 5-methyl group with 64.4, 46.4, and 27.0%, respectively. In contrast to derivatives 3, 9,
5-nitro derivative resulted in
a sharp increase in the anti- 13, and 18, introducing a nitro group, as observed for compound
inflammatory effects, as observed for compounds 5 and 9, with 23, on the phthalimide core resulted in derivative 28, with a 42.5%
57.7 and 80.9% paw oedema reduction, respectively. Interestingly, decrease in oedema inhibition. Finally, cyclic imides with 3-benzene-
converting the 3-benzenesulfonamides 2–10 (26.9–80.9% oedema sulfonamide 2–10 or oxime 11–19 fragments had the greatest lev-
inhibition) into the corresponding oxime derivatives 11–19 main- els of anti-inflammatory activity, which were almost similar to those
tained the levels of the anti-inflammatory activity (25.1–82.9% of diclofenac and celecoxib, indicating that both derivatives exerted
oedema inhibition). In the latter compounds 11–19, it was evident similar interactions at the COX receptor binding site.

614
A. A.-M. ABDEL-AZIZ ET AL.
Table 2. Results of anti-inflammatory activity of compounds 4, 9, 12, 13, 18, Table 4. Antitumor activity of trimellitimides derivatives 2–6 and 8 presented as
and celecoxib against carrageenan induced rat paw oedema in rats at three growth inhibition percentages (GI %) over 59 subpanel tumour cell lines.
graded doses.
59 cancer cell lines assay in one dose
Percentage
inhibition of paw
oedema from
control group
10.0 lM concentration: GI%
Compound
no.
Ã
Ã
PCE
Most sensitive cell lines
Compound
no.
Dose
(mg/kg)
ED50
(mg/kg)
2
3
4
5
3/59
3/59
3/59
5/59
NSC Lung Cancer (NCI-H522: 12%), Renal Cancer
(CAKI-1: 12%, UO-31: 13%).
NSC Lung Cancer (NCI-H522: 12%), CNS Cancer (SNB-
4
45
65
80
45
65
80
45
65
80
45
65
80
45
65
80
45
65
80
65.0
77.8
86.2
69.8
80.9
90.5
65.3
79.7
87.4
67.9
80.4
91.1
70.1
82.9
91.9
69.8
85.6
93.5
45.3
37.0
39.2
37.5
35.4
34.1
75: 12%), Renal Cancer (UO-31: 16%).
Leukaemia (HL-60(TB): 18%), NSC Lung Cancer (NCI-
H522: 16.0%), Renal Cancer (TK-10: 11%).
NSC Lung Cancer (HOP-92: 11%, NCI-H322M: 11%,
NCI-H522: 11%), Renal Cancer (UO-31: 17%),
Prostate Cancer (PC-3: 11%).
NSC Lung Cancer (HOP-92: 19%), CNS Cancer (SNB-
75: 12%).
9
12
6
2/59
3/59
13
8
NSC Lung Cancer (HOP-92: 11%,), CNS Cancer (SNB-
75: 13%), Renal Cancer (UO-31: 17%).
Imatinib
20/59
Leukaemia (MOLT-4: 18.0, PRMI-8226: 12.6, SR: 14.6),
NSC Lung Cancer (EKVX: 15.7, NCI-H226: 10.6,
NCI-H23: 17.1), Colon Cancer (HCT-116: 18.6, HCT-
15: 11.5, HT29: 47.1), CNS Cancer (SF-295: 15.1,
SF-539: 24.5, U251: 10.6), Melanoma (LOX IMVI:
11.6, SK-MEL-5: 22.3), Renal Cancer (A498: 13.7),
Prostate Cancer (PC-3: 10.6, DU-145: 14.4), Breast
Cancer (MDA-MB-231/ATCC: 11.2, T-47D: 18.6,
MDA-MB-468: 29.1).
18
Celecoxib
Table 3. Ulcerogenic potential of the tested compounds 4, 9,
12, 13, 18, diclofenac and celecoxib in mice .
Ã
PCE: positive cytotoxic effect which is ratio between number of cell lines with
percentage growth inhibition >10% and total number of cell lines.
Ã
Mean sum
Average
number
of ulcers
of lengths
of elongated
ulcer (mm SEM)
including leukaemia, non-small cell lung, colon, CNS, melanoma,
ovarian, renal, prostate, and breast19. The results are displayed in
Table 4 and expressed as the percentage growth inhibition (GI %)
caused by the test compounds. Based on the results, the positive
cytotoxic effects (PCE) of the tested cyclic imides 2–6 and 8 at
10 lM were 2/59–5/59, while the reference drug, imatinib, had a
PCE of 20/59 (Table 4).
Compound no.
Control
Celecoxib
Diclofenac
4
0.0
5
10
6
4
5
5
2
0.0
5.8 0.77
7.6 0.89
4.7 0.54
1.7 0.21
2.4 0.13
2.1 0.23
0.9 0.08
9
12
13
18
Ã
Significantly less than the celecoxib and diclofenac sodium
treated group using unpaired. Student’s t-test p < 0.05.
2.2.4. COX-1/2 inhibition and SAR study
The derivatives of cyclic imides 2–6, 9–15, 18, 19, 21–24, and 28,
which showed anti-inflammatory activity higher than the 30%
oedema inhibition, were subjected to COX-1 and COX-2 inhibition
assay, with the reference drug, celecoxib, using an ovine COX-1/
COX-2 assay kit (Cayman Chemicals Inc., Ann Arbour, MI). The IC50
(lM) and the selectivity indices (SI; IC50 (COX-1)/IC50 (COX-2)) are
listed in Table 53,10,11,20. Data in Table 5 show the value >387.6 as
a selectivity index (SI; IC50 (COX-1)/IC50 (COX-2)) of the reference
drug, celecoxib, with IC50 values >50/0.129 lM for COX-1/COX-2.
The COX-1/COX-2 assay indicated that cyclic imides bearing a 3-
benzenesulfonamide or 2-[4-(1-(hydroxyimino)ethyl)phenyl] frag-
ment, as observed for derivatives 2–5, 9, 11–14, 18, and 19, were
considered to be potent COX-2 inhibitors with IC50 ffi 0.15–0.90 lM
and SI ffi >333.3 to >55.6. The results of cyclic imides containing
the non-carboxylic tails 2–4, 9, 11–13, 18, and 19, were compar-
able to that of the reference drug, celecoxib (IC50 ¼ 0.129 lM;
COX-2 (SI) > 387.6). In contrast, cyclic imides containing a carbox-
ylic tail, as observed for derivatives 21–24 and 28, were non-
selective COX-1/2 inhibitors with IC50 (COX-1) ffi 10.9 ꢀ 24.8 lM and
IC50 (COX-2) ffi 22.3 ꢀ 36.3 lM and SI ffi 0.46 ꢀ 0.68.
Table 2 shows the three graded doses of the anti-inflammatory
activities of the most active cyclic imides 4, 9, 12, 13, 18,
and celecoxib.
2.2.2. Ulcerogenicity
The levels of ulcerogenic activity for the most active cyclic imides
4, 9, 12, 13, 18, along with the reference drugs, diclofenac and
celecoxib, were determined according to the reported technique
(Table 3)6a,17,18. Evidently, the phthalimide derivatives 9 and 18
had very low levels of ulcerogenic activity relative to diclofenac
and celecoxib, whereas phthalimides 4, 12, and 13 had an ulcero-
genic level similar to celecoxib and less than that of diclofenac
(Table 3).
2.2.3. Cytotoxicity
Six compounds were selected by the National Cancer Institute
(Bethesda, MD) based on structural variations to evaluate the
in vitro cytotoxicity of compounds 2–6 and 8. Thereafter, the
results were compared to those of the reference drug, imatinib, as
shown in Table 4. These compounds, which are derivatives of cyc-
lic imides bearing 3-benzenesulfonamides 2–6 and 8, were admin-
The inhibitors, cyclic imides 2–4, 9, 11–13, 18, and 19, were
the most potent and active derivatives with IC50 values of 0.26 lM
(SI > 192.3), 0.20 lM (SI > 250.0), 0.18 lM (SI > 277.8), 0.15 lM (SI
> 333.3), 0.22 lM (SI > 227.3), 0.16 lM (SI > 312.5), 0.16 lM (SI >
312.5), 0.15 lM (SI > 333.3), and 0.28 lM (SI > 178.6), respectively,
istered in single doses of 10 mM in a full NCI 59 cell line panel relative to celecoxib which had an IC50 value of 0.129 lM and SI
assay. These cell lines were obtained from nine different organs value of >387.6. The cyclic imides incorporating oximes, as

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
615
Table 5. In vitro cyclooxygenase (COX-1/COX-2) enzyme inhibition assay and cal-
(IC50 ¼ 30.0 lM) and increased COX-1 inhibitory activity (IC50
¼
culated selectivity indices.
18.2 lM and SI ¼ 0.61); (v) the tetrahydrophthalimide and phthali-
IC50 (mM)a
mide core structures, as observed for compounds 3 (IC50
¼
SIb
0.20 lM and SI > 250), 4 (IC50 ¼ 0.18 lM and SI > 277.8), 12 (IC50
¼ 0.16 lM and SI > 312.5), and 13 (IC50 ¼ 0.16 lM and SI >
312.5), had higher inhibitory activity than the succinimide deriva-
tives, as observed for compounds 2 (IC50 ¼ 0.26 lM and SI >
192.3) and 11 (IC50 ¼ 0.22 lM and SI > 227.3). These aforemen-
tioned results indicate the importance of the double bonds for
COX-2 inhibition; (vi) replacing the phthalimide fragments in com-
Compound no.
COX-1
COX-2
2
3
4
5
6
9
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
16.60
11.60
10.90
24.80
18.20
>50
0.26
0.20
0.18
0.90
4.00
0.15
3.50
0.22
0.16
0.16
0.80
3.50
0.15
0.28
30.30
25.10
22.30
36.30
30.00
0.129
>192.3
>250.0
>277.8
>55.6
>12.5
>333.3
>14.3
10
11
12
13
14
15
18
19
21
22
23
24
28
Celecoxib
>227.3 pounds 4 and 13 (IC50 ¼ 0.18 lM, and 0.16 lM, respectively), with
>312.5
>312.5
>62.5
>14.3
the naphthalimide structure in compounds 10 and 19 decreasing
the COX-2 inhibitory activity with IC50 of 3.5 lM and 0.28 lM,
respectively; (vii) substituting the phthalimide core in compounds
4 (IC50 ¼ 0.18 lM), 13 (IC50 ¼ 0.16 lM) and 23 (IC50 ¼ 19.3 lM)
>333.3
>178.6
0.55
0.46
0.49
0.68
0.61
>387.6
with the methyl moiety (CH3) at the 5-position in compounds 5,
14, and 24 resulted in a decrease in COX-2 inhibition (IC50
¼
0.9 lM, 0.8 lM, and 36.30 lM, respectively) and an increase in
COX-1 inhibition of compound 24 (IC50 ¼ 24.8 lM); (viii) replacing
the methyl fragment (CH3) in compounds 5 and 14 (IC50
¼
0.9 lM, and 0.8 lM, respectively) with the bulky tert-butyl group
(–C(CH3)3) in compounds 6 and 15 led to a decrease in COX-2
inhibition (IC50 ¼ 4.0 lM, and 3.5 lM, respectively); (ix) converting
the terminal sulphonamide or oxime fragments in compounds 3
(IC50 ¼ 0.2 lM), 4 (IC50 ¼ 0.18 lM), 12 (IC50 ¼ 0.16 lM), and 13
(IC50 ¼ 0.16 lM) into a carboxylic acid (–COOH) moiety containing
the same cyclic imide scaffold found in compounds 22 and 23
resulted in a decrease in COX-2 inhibition (IC50 ¼ 25.1 lM and
aIC50 value is the compound concentration required to produce 50% inhibition
of COX-1or COX-2 for means of two determinations using an ovine COX-1/COX-2
assay kit (catalog no. 560101, Cayman Chemicals Inc., Ann Arbour, MI) and devi-
ation from the mean is <10% of the mean value. bSelectivity index (COX-1 IC50
/
COX-2 IC50).
observed for compounds 11–14 and 19 (COX-2 (SI) >62.5ꢀ 312.5),
were more active than the corresponding cyclic imides bearing a 3-
benzenesulfonamide tail, as observed for compounds 2–5 and 10
(COX-2 (SI) ꢁ14.3ꢀ 277.8). Interestingly, compounds 9 and 18 con-
taining the 3-benzenesulfonamide and oxime tails, respectively, had
the same COX-2 (SI) value >333.3, which may be attributed to the
same binding interaction with the COX-2 putative pocket. The pres-
ence of the carboxylic moieties on the cyclic imides, as observed
for series 21–24 and 28, afforded non-selective COX-1/COX-2 inhib-
ition with a relatively high COX-1 inhibition of IC50 ffi 10.9–24.8lM)
and lower COX-2 inhibition of IC50 ffi 22.3–36.3lM). Compounds 22
and 23 had low COX-2 inhibition, with IC50 values of 25.1 and
22.3lM, respectively, and high COX-1 inhibition, with IC50 values of
22.3 lM, respectively) and increase in COX-1 inhibition (IC50
¼
11.6 lM and 10.9 lM, respectively). Briefly, the non-carboxylic cyc-
lic imide scaffolds containing the 3-benzenesulfonamide or 2-[4-
(1-(hydroxyimino)ethyl)phenyl] fragments, as observed for deriva-
tives 2–5, 9, 11–14, 18, and 19, were the most active inhibitors of
the COX-2 isozyme and some displayed higher levels of selective
COX-2 inhibition (SI ꢂ 250 ꢀ 333.3), which were comparable to
levels for celecoxib (SI ꢂ 387.6).
2.3. Molecular docking studies
11.6 and 10.9lM, respectively, compared to their 3-benzenesulfona- The molecular modelling technique was used to establish and
mide and oxime analogues 3, 4, 12, and 13, with SI values of understand the binding mode of the most bioactive com-
>250.0, 277.8, 312.5, and >312.5, respectively. Hence, the sulphona- pounds21,22. The selectivity of the cyclic imide analogues towards
mide (–SO2NH2) and oxime (–C¼NOH) fragments are important COX-2 was studied using a molecular docking protocol on the
pharmacophores for the interaction with the COX-2 binding pocket. MOE 2008.10 programme obtained from Chemical Computing
The structure-activity relationships (SAR) of the test compounds Group Inc. (Montreal, Canada)23.
2–6, 9–15, 18, 19, 21–24, and 28 in the COX-1/2 inhibition assay
Molecular docking was performed to examine the best inter-
revealed the following: (i) generally, the COX-2 isozyme was action between the most active compounds, 9 and 18, and the
potentially affected by cyclic imides 2–5, 9, 11–14, 18, and 19, COX-2 pocket binding site (Figure 2, lower panels). The crystallo-
with IC50 values ranging from 0.15 ꢀ 0.90 lM and SI ffi >333.3 to graphic binding site on the COX-2 isozyme in complex with the
>55.6 compared to celecoxib (IC50 ¼ 0.129 lM; COX-2 (SI) > SC-558 ligand, an analogue of celecoxib, was derived from Protein
387.6); (ii) the non-carboxylic cyclic imides, including the 3-benze- Data Bank (PDB code: 1CX2) (Figure 2, upper panels)4.
nesulfonamide and oxime tails 2–5, 9, 11–14, 18, and 19, were
The scoring function and the hydrogen bond formed among
potent inhibitors of the COX-2 isozyme, while the carboxylic cyclic these compounds and the surrounding amino acids were used to
imide derivatives 21–24 and 28 were very weak or non-COX-2 predict the interaction mode of compounds 9 and 18 in the puta-
inhibitors. Consequently, the sulphonamide (SO2NH2) and oxime tive active pocket of the COX-2 isozyme. Figure 2 shows the
(–C¼NOH) fragments are an essential part of COX-2 inhibition and results of the docking studies for both compounds 9 and 18,
recognition; (iii) the cyclic imides with a nitro (NO2) group at the which possess the common cyclic imide pharmacophore and a
5-position in compounds containing sulphonamide and oxime similar binding interaction, including H-bonding and hydrophobic
fragments, as observed for compounds 9 and 18, possessed the interactions within the putative binding pocket. These compounds
strongest COX-2 inhibitory activity among the tested compounds were inserted deep into the hydrophilic site of the COX-2 isozyme
(IC50 ¼ 0.15 lM and SI > 333.3); (iv) in contrast, the carboxylic cyc- where the sulphonamide and oxime groups can interact with the
lic imide substituted with a nitro (NO2) group at the 5-position, as hydrophilic pocket (His90, Gln192, and Arg513) through a network
observed for compound 28, had reduced COX-2 inhibitory activity of hydrogen bonds; such binding interactions were similar to that

616
A. A.-M. ABDEL-AZIZ ET AL.
Figure 2. 3D interactions of compounds SC-558 ligand (upper panel), 9 (lower left panel), and 18 (lower right panel) with the active site of COX-2. Hydrogen bonds
are depicted as red lines.
of SC-558 co-crystallized in the COX-2 active site4. Additionally, (3.82 Å) similar to the sulphonamide moiety in compound 9
[Figure 2, lower right panel]. Additionally, the methyl moiety of
the oxime structure interacted with amino acid Leu352 through a
non-classical H-bond (2.94 Å), while the N-phenyl moiety formed
another non-classical H-bond with Tyr355 (2.77 Å) [Figure 2, lower
right panel]. Moreover, the two carbonyl groups of cyclic imide
were held by one classical and two non-classical H-bonds with
amino acids Arg120 (2.65 Å), Ser353 (2.94 Å), and Val349 (2.81 Å)
[Figure 2, lower right panel]. Finally, the 5-nitro group of cyclic
imide formed a non-classical H-bond with Leu531 (3.08 Å). The
binding interactions described herein are typical of the pyrazolic
prototype (SC-558), which is a selective inhibitor of COX-2.
Altogether, such findings confirm the molecular design of
the reported class of anti-inflammatory 1,3-isoindoledione
the cyclic imide cores of compounds 9 and 18 were oriented at
the top of the channel, which were involved in the hydrophobic
interaction with amino acids, Ala527, Val349, Trp387, Val523, and
Leu352. The terminal sulphonamide group (–SO2NH2) was respon-
sible for the stability of the docked compound 9 through its con-
served role in the binding pocket through the formation of
suitable H-bonds with the key amino acids, Arg513 (2.87 Å), His90
(3.12 Å), Gln192 (2.78 Å), and Phe518 (3.76 Å) [Figure 2, lower left
panel]. Meanwhile, the phenyl ring of the 3-benzenesulfonamide
formed two non-classical H-bonds by binding with amino acids,
His90 (3.27 Å) and Tyr355 (2.96 Å), and an additional hydrophobic
interaction with Ser353 (3.95 Å) through CH-pi interactions. The 5-
nitro group of cyclic imide was also identified to form three H-
bonds with the amino acids Tyr385 (2.67 Å) and Ser530 (2.76 Å
and 2.72 Å), whereas one of the carbonyl group of cyclic imide
interacted with the amino acid Val523 through a non-classical
H-bond (3.20 Å) [Figure 2, lower left panel].
scaffolds10,11
.
3. Conclusions
In a similar manner, the complex generated by the docking of
compound 18 [Figure 2, lower right panel] showed that the ter-
minal oxime moiety (C¼N–OH) formed H-bonds with amino acids
The cyclic imide scaffolds bearing the non-carboxylic 3-benzene-
sulfonamide or 2-[4-(1-(hydroxyimino)ethyl)phenyl] fragments
2–10, and 11–19 and the carboxylic derivatives 21–29 were syn-
Arg513 (2.89 Å), His90 (2.95 Å), Gln192 (2.88 Å), and Phe518 thesised and evaluated for their in vivo anti-inflammatory and

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
617
ulcerogenic activities and in vitro cytotoxicity. The derivatives J ¼ 12.6 Hz), 2.48–2.51 (d, 2H, J ¼ 16.1 Hz), 3.35 (s, 2H), 5.98 (s, 2H),
revealed as the most active anti-inflammatory agents were also
subjected to an inhibition assay with the COX-1/COX-2 enzymes.
7.46–7.47 (d, 1H, J ¼ 7.0 Hz), 7.52 (s, 2H), 7.70 (s, 2H), 7.88–7.89 (d,
1H, J ¼ 6.3 Hz); 13C NMR (176 MHz, DMSO-d6): d 23.71, 40.35,
Among the tested compounds, cyclic imide derivatives bearing 3-
benzenesulfonamides (2–4, 9), acetophenone oximes (11, 12, 13),
and b-phenylalanine (18) exhibited remarkable anti-inflammatory
activities (71.2–82.9% oedema inhibition) compared to the refer-
ence drugs, celecoxib and diclofenac (85.6 and 83.4% oedema
inhibition, respectively). Cyclic imides attached to 3-benzenesulfo-
namide or oxime were stronger anti-inflammatory agents and
selective COX-2 inhibitors than the carboxylic acid derivatives con-
taining the same scaffolds. Compounds 2–4, 9, 11–13, 18, and 19
were the most potent COX-2 inhibitors (IC50 ¼ 0.26, 0.20, 0.18,
0.15, 0.22, 0.16, 0.16, 0.15, and 0.28 lM, respectively) and demon-
strated values comparable to that of celecoxib (IC50 ¼ 0.129 lM).
Some cyclic imide derivatives 2–6 and 8 were subjected to cyto-
toxic evaluation which showed weak positive cytotoxic effects
(PCE ¼ 2/59–5/59) compared to the standard drug, imatinib (PCE
¼ 20/59). From the COX-2 inhibition assay and docking results,
compounds 9 and 18 were recognised as the most active ana-
logues, with the highest recognition at the COX-2 binding site
and a correlation identified with the COX-2 selectivity indices.
124.52, 126.03, 128.23, 130.26, 130.73, 133.22, 145.35, 179.55;
C14H14N2O4S: m/z (306.3).
4.1.1.3. 3-(5-Methyl-1,3-dioxoisoindolin-2-yl)benzenesulfonamide
(5). M.P. 225–226ꢃC, 86% yield (Methanol); IR (KBr, cmꢀ1) ꢀ: 3308,
3228 (NH2), 1779, 1717 (C¼O), 1339, 1164 (O¼S¼O); 1H NMR
(700 MHz, DMSO-d6): d 2.54 (s, 3H), 7.55 (s, 2H), 7.69–7.71 (d, 1H,
J ¼ 7.8 Hz), 7.73–7.76 (q, 2H, J ¼ 7.9 Hz), 7.82 (s, 1H), 8.87–7.91 (q,
2H, J ¼ 7.8 Hz), 7.95 (s, 1H); 13C NMR (176 MHz, DMSO-d6): d 21.89,
123.95, 124.40, 124.76, 125.57, 129.34, 130.12, 130.98, 132.31,
132.82, 135.69, 145.29, 146.35, 167.19, 167.29; C15H12N2O4S: m/
z (316.4).
4.1.1.4.
3-(5-(tert-Butyl)-1,3-dioxoisoindolin-2-yl)benzenesulfona-
mide (6). M.P. 151–153ꢃC, 79% yield (Methanol); IR (KBr, cmꢀ1) ꢀ:
3354, 3264 (NH2), 1777, 1718 (C¼O), 1375, 1164 (O¼S¼O); 1H
NMR (700 MHz, DMSO-d6): d 1.39 (s, 9H), 7.57 (s, 2H), 7.69–7.70 (d,
1H, J ¼ 7.8 Hz), 7.74–7.77 (t, 1H, J ¼ 7.8 Hz), 7.91–7.95 (q, 3H,
J ¼ 7.8 Hz), 7.96–7.97 (d, 2H, J ¼ 6.4 Hz); 13C NMR (125 MHz, DMSO-
d6): d 31.26, 36.03, 120.81, 123.98, 124.83, 125.64, 129.46, 130.17,
131.09, 132.19, 132.27, 132.84, 145.28, 159.10, 167.09, 167.42;
C18H18N2O4S: m/z (358.3).
4. Experimental
4.1. Chemistry
Melting points (uncorrected) were recorded on a Barnstead 9100
Electrothermal melting apparatus (APS Water Services Corporation,
Van Nuys, CA) while the IR spectra were recorded on a FT-IR
Perkin-Elmer spectrometer (PerkinElmer Inc., Waltham, MA). The
1H NMR and 13C NMR were measured in DMSO-d6 or CDCl3, on
Bruker 700 or 500 and 176 or 125 MHz instruments, respectively
(Bruker, Billerica, MA). Chemical shifts are reported in d ppm. Mass
spectra were recorded on an Agilent 6320 Ion Trap mass spec-
trometer (Agilent Technologies, Santa Clara, CA). C, H, and N were
analysed at the Research Centre, College of Pharmacy, King Saud
University, Saudi Arabia. The results were within 0.4% of the the-
oretical values. Compounds 4, 8, 9, 12–17, and 22–24 were pre-
4.1.1.5.
3-(5,6-Dichloro-1,3-dioxoisoindolin-2-yl)benzenesulfona-
mide (7). M.P. 256–258ꢃC, 71% yield (Methanol); IR (KBr, cmꢀ1) ꢀ:
3338, 3240 (NH2), 1779, 1718 (C¼O), 1330, 1154 (O ¼ S¼O); 1H
NMR (500 MHz, DMSO-d6): d 7.50 (s, 2H), 7.65–7.66 (d, 1H,
J ¼ 8.3 Hz), 7.70–7.73 (t, 1H, J ¼ 7.9 Hz), 7.92–7.93 (d, 1H, J ¼ 7.9 Hz),
7.95–7.96 (t, 1H, J ¼ 3.5 Hz), 8.21 (s, 2H); 13C NMR (125 MHz, DMSO-
d6): d 124.69, 125.93, 125.98, 130.01, 130.58, 131.76, 132.27,
138.37, 145.44, 165.27; C14H8Cl2N2O4S: m/z (371.2).
4.1.1.6. 3-(1,3-Dioxo-1H-benzo[de]isoquinolin-2(3H)-yl)benzenesul-
fonamide (10). M.P. 241–243ꢃC, 76% yield (Methanol); IR (KBr,
cmꢀ1
)
ꢀ: 3367, 3221 (NH2), 1776, 1711 (C¼O), 1336, 1148
pared according to a previously reported procedure6b,24
.
(O¼S¼O); 1H NMR (500 MHz, DMSO-d6): d 7.39 (s, 2H), 7.52–7.53
(d, 1H, J ¼ 7.8 Hz), 7.82–7.85 (m, 2H), 8.38–8.40 (d, 2H, J ¼ 8.3 Hz),
8.43–7.45 (d, 2H, J ¼ 8.3 Hz), 8.50–8.54 (m, 3H); 13C NMR (125 MHz,
DMSO-d6): d 118.94, 122.73, 125.96, 126.99, 127.32, 127.73, 129.76,
131.33, 133.15, 134.92, 135.85, 160.77, 163.97; C18H12N2O4S: m/
z (352.3).
4.1.1. General procedure for the synthesis of 1,3-isoindole-
diones 2–10
A mixture of an equimolar amount of 3-benzenesulfonamide and
acid anhydrides (5.0 mmol) was heated under reflux for 12 h in
glacial acetic acid (15 ml) containing anhydrous sodium acetate
(0.69 g, 5.0 mmol) (Scheme 1). The reaction mixture was cooled, fil-
tered, and the obtained solid was washed with water, dried, and
re-crystallized.
4.1.2. General procedure for the synthesis of 1,3-isoindole-
dione 11–19
A mixture of 1,3-isoindolinediones (5.0 mmol) and hydroxylamine
hydrochloride (0.56 g, 8 mmol) was stirred at room temperature
for 24 h in glacial acetic acid (20 ml) (Scheme 2). The reaction mix-
ture was then filtered and the obtained solid was washed with
water, dried, and re-crystallized.
4.1.1.1. 3-(2,5-Dioxopyrrolidin-1-yl)benzenesulfonamide (2). M.P.
195–197ꢃC, 90% yield (Ethanol); IR (KBr, cmꢀ1) ꢀ: 3337, 3246 (NH2),
1696 (C¼O), 1339, 1157 (O ¼ S¼O); 1H NMR (500 MHz, DMSO-d6):
d 2.82 (s, 4H), 7.37 (s, 2H), 7.45 ꢀ 7.47 (t, 1H, J ¼ 7.9 Hz), 7.60 ꢀ 7.63
(t, 1H, J ¼ 7.9 Hz), 7.82 (s, 1H), 7.88 ꢀ 7.90 (d, 1H, J ¼ 8.0 Hz); 13C
NMR (125 MHz, DMSO-d6): d 28.82, 124.78, 125.87, 129.69, 130.46,
133.19, 145.19, 176.68; C10H10N2O4S: m/z (254.3).
4.1.2.1.
1-(4-(1-(Hydroxyimino)ethyl)phenyl)pyrrolidine-2,5-dione
(11). M.P. 184–186ꢃC, 89% yield (Methanol); IR (KBr, cmꢀ1) ꢀ: 3358
1
(OH), 1774, 1690 (C¼O), 1634 (C¼N); H NMR (700 MHz, DMSO-d6):
4.1.1.2. 3-(1,3-Dioxo-1,3,3a,4,7,7a-hexahydro-2H-isoindol-2-yl)ben-
d 2.12 (s, 3H), 2.52–2.54 (t, 2H, J ¼ 6.7 Hz), 2.57–2.59 (t, 2H,
J ¼ 6.4 Hz), 7.58–7.61 (q, 4H, J ¼ 9.5 Hz), 10.08 (s, 1H); 13C NMR
(176 MHz, DMSO-d6): d 11.83, 29.20, 31.50, 119.03, 131.94, 140.10,
zenesulfonamide (3). M.P. 159–161ꢃC, 91% yield (Ethanol); IR (KBr,
cmꢀ1
)
ꢀ: 3293, 3125 (NH2), 1773, 1699 (C¼O), 1341, 1160
(O¼S¼O); 1H NMR (700 MHz, DMSO-d6): d 2.30–2.32 (d, 2H, 152.95, 170.70, 174.32; C12H12N2O3: m/z (232.1).

618
A. A.-M. ABDEL-AZIZ ET AL.
4.1.2.2. 2-(4-(1-(Hydroxyimino)ethyl)phenyl)-5-nitroisoindoline-1,3- 4.1.3.4. 3-Phenyl-3-(4,5,6,7-tetrachloro-1,3-dioxoisoindolin-2-yl)pro-
dione (18). M.P. 269–271ꢃC, 92% yield (Ethanol); IR (KBr, cmꢀ1) ꢀ:
3428 (OH), 1737, 1699 (C¼O), 1645 (C¼N); 1H NMR (700 MHz,
DMSO-d6): d 2.21 (s, 3H), 7.47–7.48 (d, 2H, J ¼ 8.4 Hz), 7.81–7.82 (d,
2H, J ¼ 8.3 Hz), 8.12–8.14 (t, 1H, J ¼ 7.8 Hz), 8.26–8.27 (d, 1H,
J ¼ 7.4 Hz), 8.34–8.35 (d, 1H, J ¼ 7.1 Hz), 11.39 (s, 1H); 13C NMR
(176 MHz, DMSO-d6): d 12.04, 123.37, 126.53, 127.55, 127.79,
128.88, 132.06, 134.01, 136.90, 137.45, 144.98, 152.95, 163.02,
165.60; C16H11N3O5: m/z (325.1).
panoic acid (27). M.P. 280–282ꢃC, 93% yield (Methanol); IR (KBr,
cmꢀ1) ꢀ: 2870 (OH), 1741, 1715 (C¼O); 1H NMR (500 MHz, DMSO-
d6): d 3.28–3.33 (t, 1H, J ¼ 12.0 Hz), 3.49–3.53 (dd, 1H, J ¼ 4.9,
14.3 Hz), 5.15–5.18 (dd, 1H, J ¼ 4.7, 11.2 Hz), 7.13–7.21 (m, 5H),
13.47 (s, 1H); 13C NMR (125 MHz, CDCl3): d 34.25, 54.18, 127.09,
127.48, 128.87, 129.11, 129.15, 137.53, 139.62, 163.03, 169.90;
C17H9Cl4NO4: m/z (433.1).
4.1.3.5. 3-(5-Nitro-1,3-dioxoisoindolin-2-yl)-3-phenylpropanoic acid
(28). M.P.(0).191–193ꢃC, 83% yield (Ethanol); IR (KBr, cmꢀ1) ꢀ: 2910
4.1.2.3. 2-(4-(1-(Hydroxyimino)ethyl)phenyl)-1H-benzo[de]isoquino-
line-1,3(2H)-dione (19). M.P. >300ꢃC, 92% yield (Ethanol); Yield,
87%; IR (KBr, cmꢀ1) ꢀ: 3090 (OH), 1775, 1716 (C¼O), 1649 (C¼N);
1H NMR (500 MHz, DMSO-d6): d 2.22 (s, 3H), 7.29–7.31 (t, 2H,
J ¼ 6.5 Hz), 7.76–7.77 (t, 2H, J ¼ 6.5 Hz), 7.81–7.84 (t, 2H, J ¼ 7.5 Hz),
8.38–8.40 (d, 2H, J ¼ 8.5 Hz), 8.51–8.52 (d, 2H, J ¼ 6.5 Hz), 11.14 (s,
1H); 13C NMR (125 MHz, DMSO-d6): d 11.93, 122.82, 126.43, 127.29,
129.13, 129.53, 130.46, 131.27, 134.75, 136.01, 137.72, 146.22,
152.80, 162.82, 164.02; C20H14N2O3: m/z (330.1).
1
(OH), 1745, 1715 (C¼O); H NMR (700 MHz, DMSO-d6): d 3.29–3.34
(dd, 1H, J ¼ 4.0, 16.0 Hz), 3.49–3.52 (dd, 1H, J ¼ 6.9, 13.0 Hz),
5.15–5.18 (dd, 1H, J ¼ 6.9, 16.0 Hz), 7.13–7.21 (m, 5H), 8.07–8.10 (t,
1H, J ¼ 11.0 Hz), 8.16–8.18 (d, 1H, J ¼ 10.2 Hz), 8.31–8.33 (d, 1H,
J ¼ 11.3 Hz); 13C NMR (176 MHz, DMSO-d6): d 34.30, 54.03, 122.39,
127.12, 127.88, 128.63, 128.85, 129.20, 129.50, 129.66, 132.76,
137.50, 137.62, 144.88, 162.86, 165.51, 169.69, 170.07; C17H12N2O6:
m/z (340.1).
4.1.3.6.
3-(1,3-Dioxo-1H-benzo[de]isoquinolin-2(3H)-yl)-3-phenyl-
propanoic acid (29). M.P. 151–153ꢃC, 66% yield (Methanol); IR
4.1.3. General procedure for the synthesis of 1,3-isoindole-
dione 21–29
(KBr, cmꢀ1) ꢀ: 2930 (OH), 1708, 1667 (C¼O); 1H NMR (700 MHz,
DMSO-d6):
d
2.92–2.95 (t, 1H, J ¼ 7.7 Hz), 4.24–4.26 (t, 1H,
A mixture of an equimolar amount of b-phenylalanine (0.83 gm,
5.0 mmol) and acid anhydrides (5.0 mmol) was heated under reflux
for 24 h in glacial acetic acid (20 ml) containing anhydrous sodium
acetate (0.69 g, 5.0 mmol) (Scheme 3). The reaction mixture was
cooled and filtered, and the obtained solid was washed with
water, dried, and re-crystallized.
J ¼ 7.6 Hz), 4.95–4.97 (dd, 1H, J ¼ 7.6, 14.5 Hz), 7.28–7.31 (q, 3H,
J ¼ 7.4 Hz), 7.85–7.87 (t, 1H, J ¼ 7.5 Hz), 7.90–7.93 (q, 2H, J ¼ 7.6 Hz),
8.44 (s, 2H), 8.47-8.48 (s, 1H, J ¼ 6.9 Hz), 8.51–8.54 (t, 2H,
J ¼ 7.5 Hz); 13C NMR (176 MHz, DMSO-d6): d 33.96, 34.82, 54.73,
119.44, 122.42, 127.69, 128.02, 128.54, 128.95, 129.10, 129.40,
131.20, 132.95, 134.83, 135.19, 135.87, 139.21, 161.18, 163.51,
163.74; C21H15NO4: m/z (345.1).
4.1.3.1. 3-(2,5-Dioxopyrrolidin-1-yl)-3-phenylpropanoic acid (21).
M.P. 259–261ꢃC, 93% yield (Ethanol); IR (KBr, cmꢀ1) ꢀ: 3111 (OH),
1743, 1702 (C¼O); 1H NMR (700 MHz, DMSO-d6): d 2.44–2.49 (m,
2H), 2.52–2.57 (m, 2H), 3.24–3.27 (dd, 1H, J ¼ 11.9, 14.0 Hz),
3.37–3.39 (dd, 1H, J ¼ 4.5, 14.1 Hz), 4.59–4.61 (dd, 1H, J ¼ 4.5,
11.7 Hz), 7.09–7.10 (d, 2H, J ¼ 7.3 Hz), 7.15–7.17 (t, 1H, J ¼ 7.3 Hz),
7.22–7.24 (t, 2H, J ¼ 7.5 Hz); 13C NMR (176 MHz, DMSO-d6): d 28.02,
34.06, 55.53, 126.60, 128.68, 129.05, 139.32, 170.38, 177.55;
C13H13NO4: m/z (247.2).
4.2. Biological evaluation
4.2.1. Anti-inflammatory screening
Anti-inflammatory assessment of the newly synthesised com-
pounds was carried out using an in vivo rat carrageenan-induced
foot paw oedema model, as reported previously16,17. Compounds
6, 7, 8, 10, 11, 18, diclofenac, and celecoxib were tested at three
different doses and their ED50 was determined.
4.1.3.2.
3-(5-(tert-Butyl)-1,3-dioxoisoindolin-2-yl)-3-phenylpropa-
noic acid (25). M.P.(0).119–121ꢃC, 76% yield (Methanol); IR (KBr,
cmꢀ1) ꢀ: 3219 (OH), 1739, 1703 (C¼O); 1H NMR (700 MHz, DMSO-
d6): d 1.32 (s, 9H), 3.43–3.47 (t, 1H, J ¼ 12.8 Hz), 3.52–3.55 (dd, 1H,
J ¼ 4.0, 14.4 Hz), 4.57–4.60 (dd, 1H, J ¼ 4.0, 12.5 Hz), 7.06–7.08 (t,
1H, J ¼ 7.3 Hz), 7.10–7.11 (d, 2H, J ¼ 7.3 Hz), 7.15–7.18 (t, 2H,
J ¼ 7.6 Hz), 7.67–7.69 (d, 1H, J ¼ 7.8 Hz), 7.72 (s, 1H), 7.79–7.81 (dd,
1H, J ¼ 1.5, 7.9 Hz); 13C NMR (176 MHz, DMSO-d6): d 31.22, 35.35,
35.83, 57.41, 119.93, 123.04, 126.28, 128.65, 128.80, 129.60, 131.50,
132.38, 140.75, 158.31, 168.60, 168.89, 174.40; C21H21NO4: m/
z (351.2).
4.2.2. Ulcerogenicity measurement
Ulcerogenicity was evaluated according to a previously reported
method6a,17,18. The number and total length of the ulcers for each
animal were measured and their averages were calculated and
used as the ulcer indices.
4.2.3. In vitro cyclooxygenase (COX) inhibition assay
To determine the relative ability of the test compounds and refer-
ence drugs to inhibit COX-1/COX-2 isozymes, we used the colori-
metric COX (ovine) inhibitor screening assay kit (Cayman
Chemicals Inc., Ann Arbour, MI), according to the manufacturer’s
instructions3,10,11,20
.
4.1.3.3. 3-(5,6-Dichloro-1,3-dioxoisoindolin-2-yl)-3-phenylpropanoic
acid (26). M.P. 205–207ꢃC, 89% yield (Ethanol); IR (KBr, cmꢀ1) ꢀ:
2890 (OH), 1742, 1717 (C¼O); 1H NMR (500 MHz, DMSO-d6): d
3.28–3.33 (dd, 1H, J ¼ 12.0, 14.0 Hz), 3.47–3.51 (dd, 1H, J ¼ 4.7,
14.1 Hz), 5.12–5.15 (dd, 1H, J ¼ 4.8, 11.7 Hz), 7.11–7.19 (m, 5H), 8.16
(s, 2H), 13.44 (s, 1H); 13C NMR (125 MHz, DMSO-d6): d 34.33, 53.86,
126.19, 127.09, 128.82, 129.17, 130.81, 137.58, 138.56, 165.67,
170.17; C17H11Cl2NO4: m/z (364.2).
4.3. Docking methodology
Molecular modelling studies were performed using the 2007.09
software from Chemical Computing Group Inc. (Montreal, Canada).
The docking protocol was similar to that mentioned in our previ-
ous report19.

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
619
inhibition activities and molecular docking study of pyrazo-
line derivatives. Bioorg Med Chem 2016;24:2032–42. (d) El-
Sayed MA-A, Abdel-Aziz NI, Abdel-Aziz AA-M, et al.
Synthesis, biological evaluation and molecular modeling
study of pyrazole and pyrazoline derivatives as selective
COX-2 inhibitors and anti-inflammatory agents. Part 2.
Bioorg Med Chem 2012;20:3306–16.
Acknowledgements
The authors thank Ahmad Abbas at Department of
Pharmacognosy, Faculty of Pharmacy, Mansoura University, Egypt
for performing in vivo biological activity.
Disclosure statement
4. (a) Penning TD, Talley JJ, Bertenshaw SR, et al. Synthesis and
biological evaluation of the 1,5-diarylpyrazole class of cyclo-
oxygenase-2 inhibitors: identification of 4-[5-(4-methyl-
phenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfona-
mide (SC-58635, Celecoxib). J Med Chem 1997;40:1347–65.
(b) Kurumbail RG, Stevens AM, Gierse JK, et al. Structural
basis for selective inhibition of cyclooxygenase-2 by anti-
inflammatory agents. Nature 1996;384:644–8. (c) Michaux C,
Charlier C. Structural approach for COX-2 inhibition. Mini
Rev Med Chem 2004;4:603–15.
No potential conflict of interest was reported by the author(s).
Funding
The authors acknowledge financial support from the Researchers
Supporting Project number (RSP-2019/40), King Saud University,
Riyadh, Saudi Arabia.
5. (a) Ghosh N, Chaki R, Mandal V, Mandal SC. COX-2 as a tar-
get for cancer chemotherapy. Pharmacol Rep 2010;62:
233–44. (b) Dai ZJ, Ma XB, Kang HF, et al. Antitumor activity
of the selective cyclooxygenase-2 inhibitor, celecoxib, on
breast cancer in Vitro and in Vivo. Cancer Cell Int. 2012;12:
53.
ORCID
Alaa A.-M. Abdel-Aziz
Abdulrahman A. Almehizia
8711-3873
http://orcid.org/0000-0002-3362-9337
http://orcid.org/0000-0001-
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260–8. (b) Abdel-Aziz AA-M, El-Azab AS, Abu El-Enin MA,
et al. Synthesis of novel isoindoline-1,3-dione-based oximes
and benzenesulfonamide hydrazones as selective inhibitors
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2018;80:706–13. (c) Abdel-Aziz AA-M, El-Azab AS, Ekinci D,
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Chem 2015;30:81–4. (d) El-Azab AS, Abdel-Aziz AA-M, Ayyad
RR, et al. Inhibition of carbonic anhydrase isoforms I, II, IV,
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