Synthesis, biological evaluation and molecular docking of novel series of spiro [(2H,3H) quinazoline-2,1′- cyclohexan]-4(1H)- one derivatives as anti-inflammatory and analgesic agents

Article abstract of DOI:10.1016/j.ejmech.2009.12.078

Three series of Spiro [(2H,3H) quinazoline-2,1′-cyclohexan]-4(1H)-one derivatives have been synthesized. Some of the novel quinazolinone derivatives IIe, VIIIc, XIc, XIIb, XIIc, XVIb showed considerable potent anti-inflammatory and analgesic activity of s

Full text of DOI:10.1016/j.ejmech.2009.12.078

European Journal of Medicinal Chemistry 45 (2010) 2117–2131
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Synthesis, biological evaluation and molecular docking of novel series
of spiro [(2H,3H) quinazoline-2,10- cyclohexan]-4(1H)- one derivatives
as anti-inflammatory and analgesic agents
,
c
b
K.M. Amin a, M.M. Kamel b, M.M. Anwar b , M. Khedr , Y.M. Syam
*
a Depatment of Therapeutical Chemistry, Faculty of Pharmacy, Cairo University, Egypt
b Department of Therapeutical Chemistry, National Research Centre, Dokki, Cairo, Egypt
c Welsh School of Pharmacy, Cardiff university, UK
a r t i c l e i n f o
a b s t r a c t
Three series of Spiro [(2H,3H) quinazoline-2,10-cyclohexan]-4(1H)-one derivatives have been synthe-
sized. Some of the novel quinazolinone derivatives IIe, VIIIc, XIc, XIIb, XIIc, XVIb showed considerable
potent anti-inflammatory and analgesic activity of superior G.I.T. safety profile in experimental rats in
comparing to indomethacin and tramadol as reference drugs. Docking study into COX-2 has been made
for derivatives of highest anti-inflammatory activity. The compound XVIb showed the nearest RMSD
value to that of indomethacin.
Article history:
Received 29 September 2009
Received in revised form
24 November 2009
Accepted 23 December 2009
Available online 21 January 2010
Ó 2010 Elsevier Masson SAS. All rights reserved.
Keywords:
Spiro [(2H,3H) quinazoline-2,10-
cyclohexan]-4(1H)-one
Antiinflammatory
Analgesic
Ulcerogenic effect
Molecular docking
Cyclooxygenase-2
1. Introduction
bearing quinazolinone nucleus. Also, it has been reported that
substitution pattern by different aryl or heteroaryl moieties at 2/3
It is well known that non steroidal anti-inflammatory drugs
(NSAIDs) are associated with several side effects such as gastro-
intestinal mucosal damage, bleeding, intolerance and renal
toxicity [1,2]. Production of safer and more active NSAIDs and
analgesic drugs is still needed. Quinazoline and quinazolinone
nuclei have drawn a great attention due to their wide range of
chemotherapeutic activities including antiviral [3], antibacterial
[4,5], antifungal [6,7], antimalarial [8], anticancer [9–11], antihy-
pertensive [12], diuretic [13,14], inhibitors of derived growth
factor receptor phosphorylation [15], anticonvulsant [16], ghrelin
receptor antagonists [17], anti-inflammatory, analgesic and COX-II
inhibitors [18–20]. Additionally, different known anti-inflamma-
tory drugs such as: Proquazone, 1-isopropyl-7-methyl-4-
phenylquinazoline-2(1H)-one(1) [21], Fluoroquazone, 4-(4-fluo-
rophenyl)-7-methyl-1-propan-2-yl-quinazoline-2-one (2) [22,23],
(Tryptanthrin) indolo [2,1-b] quinazoline alkaloid (3) [24] are
position of quinazoline nucleus markedly influences the anti-
inflammatory activity [25]. On the other hand, benzene sulfon-
amides [26], 2-oxo(imino)pyridines [27,28], pyrazoles [29–31],
pyrimidines [32], imidazoles [33] and thiazolidinones [34–36] are
other important pharmacodynamic heterocyclic nuclei which
when incorporated into different heterocyclic templates, have
been reported to possess potent anti-inflammatory activity. The
enhanced overall lipophilic characteristics of the target
compounds could favor their selectivity towards COX-2 enzyme
over COX-1 leading to increase of GIT safety margin [26]. Based on
the above observations and in continuation of our anti-inflam-
matory and analgesic drug research program, it was of interest to
synthesize
a novel series of quinazolinone derivatives with
structure modifications involving incorporation of the above
mentioned heterocyclic moieties at 3rd position and a spiro
cyclohexane moiety at 2nd position of quinazolinone moiety as
a trial to obtain safer and potent anti-inflammatory and analgesic
agents. The ulcerogenic activity of the active compounds was
determined.
* Corresponding author. Tel.: þ20 123956970.
E-mail address: manal.hasan52@yahoo.com (M.M. Anwar).
0223-5234/$ – see front matter Ó 2010 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejmech.2009.12.078

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K.M. Amin et al. / European Journal of Medicinal Chemistry 45 (2010) 2117–2131
respectively, while their reaction with thiourea in boiling ethanol in
O
the presence of potassium hydroxide, gave the corresponding
pyrimidine-2-thione derivatives XIa,b, according to reported
methods [42].
N
N
N
On the other hand, it is well known that several pyridone and
imino pyridine derivatives are of chemotherapeutic importance
[43]. In this study, 2(1H) pyridones XIa–c were obtained following
the reported procedure [43,44] by heating a mixture of equimolar
amounts of the acetyl derivative V with the appropriate aromatic
aldehyde namely: benzaldehyde, p-flourobenzaldehyde and/or
2-hydroxy benzaldehyde and ethyl cyanoacetate in the presence of
excess ammonium acetate in n-butanol. Upon applying the same
procedure, using malononitrile instead of ethyl cyanoacetate, the
target 2(1H) iminopyridines XIIa–c were obtained (Scheme 2).
Additionally, as a structure variation of the para substitution of
the phenyl ring at the 3-position of the quinazolinone moiety, the
ester derivative XIII was prepared by condensation of benzoxazine
derivative I with ethyl-p-aminobenzoate in refluxing glacial acetic
acid in the presence of anhydrous sodium acetate. The preparation
of acid hydrazide XIV was achieved through hydrinolysis of the
ester precursor XIII with excess hydrazine hydrate. Treatment of
the derivative XIV with p-methoxybenzaldehyde and/or 2-thio-
phenaldehyde in absolute ethanol containing few drops of acetic
acid afforded the desired Schiff’s base derivatives XVa,b in
moderate yields. Furthermore, the intermediate Schiff bases XVa,b
were cyclized into the corresponding thiazolidine-4-one deriva-
tives XVIa,b by their cyclocondensation reaction by thioglycolic
acid in refluxing dry benzene [45]. Synthesis of unsymmetrical
ureas and/or thioureas XVIIa–c was achieved via the reaction of
the hydrazide derivative XIV with cyclohexyl isocyanate and
cyclohexyl and/or phenyl isothiocyanate in dry benzene containing
few drops of triethylamine. Finally, preparation of the imidazoli-
dinone derivatives XVIIIa–c were achieved by the reaction of the
derivatives XVIIa–c with chloroacetic acid in the presence of few
drops of pyridine, while their reaction with the bielectrophilic
species, malonic acid, it gave the corresponding, 4,6-dioxo-2-
thio(oxo)pyrimidin e.g. derivatives XIXa–c in moderate yields.
(Scheme 3).
F
N
O
2
1
N
N
3
2. Chemistry
The desired spiro [(2H, 3H)-3(4-substituted aminosulfonyl
phenyl) quinazoline-2,10-cyclohexan]-4(1H) ones (IIa–e) were
obtained by aromatic aminolysis [37] of the known benzoxazine
derivative, namely: spiro [2H-3,1-benzoxazine-2,10-cyclohexan]-
4(1H)-one (I) [38,39], upon reaction with the appropriate sulfa
drugs, namely: sulfanilamide, sulfaguanidine, sulfamethoxazole,
sulfapyridine and sulfadiazine in refluxing glacial acetic acid in
the presence of anhydrous sodium acetate. The sulfonyl urea or
(thiourea) derivatives IIIa–c were prepared in good yields by the
reaction of aminosulfonylphenyl derivative IIa with ethyl iso-
thiocyanate, cyclohexyl and phenyl isocyanates in refluxing dry
acetone in the presence of anhydrous potassium carbonate.
N-alkylation of the derivative IIa was carried out by its reaction
with ethyl iodide in refluxing ethanol in the presence of anhydrous
potassium hydroxide to give the corresponding N-ethyl-
lquinazolinone derivative, namely, spiro [(2H,3H)-3(4-amino-
sulfonylphenyl)-1-ethylquinazoline-2,10-cyclohexan]-4(1H)one
(IV) (Scheme 1).
3. Results and discussion
Also, the starting benzoxazine derivative I was allowed to
condense with p-amino acetophenone in glacial acetic acid in the
presence of anhydrous sodium acetate to give the corresponding
key intermediate, 3-(p-acetyl phenyl)quinazolin-4(1H)one deriva-
tive V. Claisen–Schmidt condensation of the acetyl derivative V
with different aldehydes namely, benzaldehyde, N,N dimethyl-
aminobenzaldehyde and\or furan-2-carboxaldehyde in 5% ethanolic
3.1. Biological evaluation
3.1.1. Antiinflammatory effect
The anti-inflammatory activity of nineteen of the newly
synthesized compounds: IIa, IIe, IIIa, V, VIc,VIId, VIIIc, VIIId, XIb,
XIc, XIIb, XIIc ,XIII, XIV, XVa, XVIb, XVIIb, XVIIIb, XIXb were
evaluated by applying carrageenan-induced paw oedema bioassay
in rats [46] using indomethacin as a reference standard. Results
were expressed as mean ꢀ S.E. Difference between vehicle control
and treatment groups were tested using one way ANOVA followed
by the least significant difference (L.S.D.). Methods of statistical
analysis were done according to Armitage et al. [47].
According to Table 1, administration of many of tested
compounds 60 min prior to carrageenan injection at dose of 9 mg/kgb
wt caused significant inhibition of paw oedema response.
Compounds IIe, VIIIc, XIIc and XVIb caused significant decrease in
paw oedeama after 2, 3, 4 h after drug administration, while XIc,
XIII and XVIIIb gave their response after 2 h of administration and
continued to the third hour. Compounds XIb and XIIb showed the
effect only after 2 h but compounds VIId, XIV significantly
decreased the paw oedeama after 4 h post administration. On the
other hand compounds IIa, IIIa, V, VIc, VIIId, XVa, XVIIb and XIXb
were inactive towards carrageenan-induced oedema in comparison
to the standard reference indomethacin which markedly and
sodium hydroxide solution afforded the corresponding a,b-unsat-
urated ketones (chalcones) VIa–c respectively. These chalcones are
considered to be useful intermediates in several cyclization
reactions to produce different types of heterocyclic compounds of
diverse biological importance, according to the reactants used and
the reactions conditions [40,41].
Cyclocondensation of the unsaturated ketones VIb,c by hydra-
zine hydrate in absolute ethanol afforded the corresponding pyr-
azoline derivatives VIIa,b, but when the reaction was carried out in
glacial acetic acid, the N-acetyl pyrazoline derivatives VIIc,d were
obtained.
Also, cyclocondensation of the key ketones VIb,c by methyl
hydrazine and/or phenyl hydrazine in absolute ethanol furnished
the target N-methyl and/or N-phenyl pyrazoline derivatives VIIIa–
d. Further, when a,b-unsaturated ketones VIb,c were allowed to
react with hydroxylamine hydrochloride in ethanolic sodium
hydroxide solution, afforded the corresponding isoxazolines IXa,b,

K.M. Amin et al. / European Journal of Medicinal Chemistry 45 (2010) 2117–2131
2119
O
COOH
+
NH2
Re
flux2
hour
s
H
N
O
O
I
Different
Sulfa drugs
IIa, R= -H
IIb,R
=
-C-NH2
NH
N
IIc,R
=
H
N
CH3
O
N
IId,R
=
=
SO2NHR
N
N
N
IIe,R
IIa-e
O
R-N=C=X
Dry acetone
CH3CH2
IIa
I
IIa
CH2CH3
N
H
N
SO2NHR
N
N
SO2NHCR
X
O
O
III
IV
IIIa X=S
IIIb X=O
R= C2H5
R=
IIIc X=O
R=
Scheme 1.
significantly inhibited the paw oedema after 2, 3, 4 h of carrageenan
injection. Thus, compounds IIe, VIId, VIIIc, XIb, XIc, XIIb, XIIc, XIII,
XIV, XVIb and XVIIIb have good anti-inflammatory activity and
compound XVIb was the most potent derivative. Results are illus-
trated by Fig. 1.
exhibited the analgesic effect after 1 h of administration only.
Compounds VIIId, XIb, XIII, XVIIIb have no analgesic activity in
comparison to the base line of the same group 1 and 2 h post
administration.
Thus, it can be concluded that, compounds IIa, IIe, IIIa, V, VIc,
VIId, VIIIc, XIc, XIIb, XIIc, XIV, XVa, XVIb, XVIIb and XIXb have
significant analgesic activity and compound XIc is the most potent
one. Results are illustrated by Fig. 2.
3.1.2. Analgesic activity
The analgesic activity of the above mentioned nineteen deriv-
atives was also evaluated by applying Hot plate test [48] using
tramadol as a standard reference. Results were expressed as
mean ꢀ S.E. Difference between vehicle control and treatment
groups were tested using one way ANOVA followed by the least
significant difference (L.S.D.). Methods of statistical analysis were
done according to Armitage et al. [47].
According to Table 2, compounds IIe, IIIa, XIc, XIIb, XVIb,
showed significant analgesic activity higher than that obtained by
Tramadol 1 h and 2 h post administration.
While compounds IIa, V, VIc, XVIIb, XIXb exhibited equipotent
analgesic effect or slightly less than that of Tramadol after 1 and 2 h
of their administration. Compounds VIIIc, XIIc and XVa exhibited
significant analgesic activity higher than or slightly equipotent to
Tramadol only after 2 h of administration. Compounds VIId and XIV
3.1.3. Ulcerogenic effect
The ulcerogenic effect of the most active anti-inflammatory and
analgesic derivatives: IIe, VIIIc, XIc, XIIb, XIIc, XVIb was evaluated
[49]. According to Table 3, it has been found that compounds IIe,
XIIb , XIIc, XVIb have very little ulcerogenic effect with better
safety margin in comparison to indomethacin. Interestingly,
compound XIc exhibited no ulcerogenic effect in all of the experi-
mental animals. On the other hand compound VIIIc resulted in
ulcer lesions in many of the experimental rats.
Therefore, the potential medicinal value of these compounds as
anti-inflammatory and analgesic agents, that they have better
safety margin than indomethacin on gastric mucosa. Results are
illustrated by Fig. 3.

2120
K.M. Amin et al. / European Journal of Medicinal Chemistry 45 (2010) 2117–2131
NH2
H
N
H
N
reflux in acetic
O
+
N
COCH3
O
COCH3
I
O
V
ArCHO
ArCHO
Diff., aldhydes
10%NaOH
NCCH2CN
NC-COOCH2CH3
CH3COONH4
CH3COONH4
Ar
Ar
O
XIa, Ar=
XIIa, Ar=
CN
CN
NH
Q
Q
VIa
Q
Ar
XIb, Ar
CNCH2COOET
CH3COONH4
F
=
O
F
XIIb, Ar
=
N
H
N
H
VIa-c
XIIa-c
XIa-c
XIc, Ar=
HO
XIIc, Ar=
VIa, Ar= Ph
HO
N
VIb, Ar=
VIc, Ar=
O
R
N
N
N
O
Q
Q
Ar
N
Ar
VIIIa-d
IXa-b
Ar=
IXa,
N
N
VIIIa, R= CH3, Ar=
IXb,
Ar=
VIIIb, R=CH3, Ar=
VIIIc, R= Ph , Ar=
O
O
O
S
N
N
R
VIIId, R= Ph,
N
Ar=
Q
NH
Ar
Q
Ar
VIIa-d
Xa-b
Ar=
VIIa, R= H,
VIIb, R=H,
N
N
Ar=
Ar=
Xa,
N
Xb,
Ar=
O
O
VIIc, R=COCH3, Ar=
VIId, R=COCH
3 ,Ar=
O
H
N
Q=
N
O
Scheme 2.
3.2. Molecular docking study
3.2.1. Aim of work
fitting and closure to the main residues. As we can see in the
following Fig. 6, the most active compound XVIb which has the
best anti-inflammatory action was found to be the best in fitting,
and has the best docking score as well and we can see the possible
interactions that formed with Arg 120 and Tyr 355 and Ser 530
(Figs. 7–9).
The aim of this work was to study the crystal structure of COX-2
and to rationalize the obtained biological data and explain the
possible interactions that might take place between the tested
derivatives and COX-2 enzyme in comparing to IndomethacinÒ in
order to obtain the anti-inflammatory effect. First of all the main
interactions of IndomethacinÒ with COX-2 were determined and
we found that the main residue is Tyr 355 (–OH group) forming
interactions with the carboxylate group of Indomethacin in
a distance equal to 1.03 Ao. From the following Fig. 4, we can also
determine the other residues present in the pocket and can be
involved in the interactions such as Arg 120, Leu 531, Val 349, and
Ser 353 (Fig. 5).
3.2.3. Measurement of the affinities
The affinity of any small molecule can be considered as a unique
tool in the field of drug design. As there is a relationship between
the affinity of organic molecules and the free binding energy and
this can contribute in prediction and interpretation of the activity of
organic compounds toward a specific target protein. Here we used
Autodock Vina program [51,52] for further docking and to obtain
the affinities and RMSD of compounds that have shown anti-
inflammatory effect and to compare these data with that was
obtained from Indomethacin docking. The following Table 4 illus-
trates these results.
3.2.2. AutoDock binding affinities of the synthesized compounds
into COX-2
The crystal structure was downloaded (pdb code: 4COX) and
our compounds were docked into the active site using plants
software [50]. Compounds (IIe, VIId, VIIIc, XIb, XIc, XIIb, XIIc ,XIII,
XIV, XVIb, XVIIIb) were ranked after docking according to their
docking scores and were visualised inside the pocket to view their
All compounds were saved as pdbqt format and docking with
Autodock Vina was performed according to the specified condition
in which the grid box was adjusted to have center_x ¼ 24.15,
center_y ¼ 22.8, and center_z ¼ 12.9 by these centres the pocket
with the main residues were involved inside the box.

K.M. Amin et al. / European Journal of Medicinal Chemistry 45 (2010) 2117–2131
2121
NH2
H
N
H
N
refiux in ACOH
O
N
O
I
COOC2H5
O
XIII
COOC2H5
NH2NH2
Q
CONHNH2
XIV
ArCHO
R-N=C=X
X
O
O
Q
N
H
R
N
H
N
H
Q
NHN CHAr
XVIIa-c
XVa,b
HSCH2COOH
CH2(COOH)2
ClCH2COOH
R
Ar
N
X
O
X
CONH
S
Q
N
N
R
N
CONH
N
CONH
Q
Q
O
O
O
XIXa-c
XVIa,b
XVIIIa-c
OCH3
a, Ar=
a, R=
b, R=
c, R=
X=
X=
X=
O
S
S
b, Ar=
S
H
N
Q =
N
O
Scheme 3.
3.3. Results and discussion
formation. Ser 530 interacts with its –OH group first, because its
–COOH is out of the specified grid box and its –NH2 group just at the
margins of the grid box so, the chance for hydroxyl group was higher
for interaction. Second, the –NH2 group of Ser 530 is of a different
planefromthatof–OHwhichisdirectedtowardthesite ofactionthat
was selected for docking. Thus compound XVIb has the best docking
score with plants and good results with Autodock vina comparing to
Indomethacin docking results with both programs.
Compound IIe was able to interact by its sulfonyl group forming
hydrogen bond with –OH group of Tyr 385 (3.49 Ao)(Fig. 10). Other
compounds containing –C]O group in their side chains such as
XIV(Fig. 11) and XIII (ester –C]O) had possible hydrogen bond
formation with –OH of Tyr 385 with measured distances 2.52 Ao,
and 2.76 Ao respectively.
The cyano group in XIb was observed to have an interaction
with Tyr 385 with 2.46 Ao (Fig. 12). While other compounds such as
VIId, VIIIc, XIc, XIIb and XIIc did not show any observed bonding or
interactions. Finally, the –C]O group of thioxo-imidazolidinone
enabled compound XVIIIb to form hydrogen bond with –OH of Tyr
355 at distance equal 1.91 Ao.
Autodock vina allows the flexible docking of ligands into its site of
action. It has the ability to use all the rotatable bonds of the ligands to
give a number of conformations from which the best mode could be
achieved. In the analysis of docking results we tried to find a corre-
lationbetween thebiological results anddockingstudies. Compound
XVIb which has the highest anti-inflammatory effect has the nearest
RMSD value to that of Indomethacin. While all compounds have
nearly the same affinity toward the target, we can use RSMD as a tool
to compare between them, and this can give us a reasonable
description for its good anti-inflammatory activity. It was found that
compound XVIb has two best conformations in which it can interact
with Arg 120 –NH2 group and Tyr 355 –OH group (Fig. 6A) and with
Ser 355 –OH group (Fig. 6B). The distance between –CO (benzamide
moiety) in XVIb and –NH2 group of Arg 120 was found to be 2.3 Ao
while the distance between the –CO (4-oxo-1,3-thiazolidine) in XVIb
and –OH group of Tyr 355 was found to be 1.9 Ao while that between
–CO of quinazoline ring and –OH group of Ser 530 was found to be
1.92 Ao which means good distances for interactions and H-bond

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K.M. Amin et al. / European Journal of Medicinal Chemistry 45 (2010) 2117–2131
Table 1
Table 2
Antiinflammatory effect.
Analgesic effect.
Groups
Oedeama volume (ml)
Group
Reaction time (sec.)
1 h
2 h
3 h
4 h
Basal
1 h
2 h
Control
IIa
IIe
IIIa
V
VIc
VIId
VIIIc
VIIId
XIb
XIc
XIIb
XIIc
XIII
XIV
57.3 ꢀ 6.8
57.7 ꢀ 5.0
60.4 ꢀ 7.1
67.5 ꢀ 7.7
54.6 ꢀ 5.8
60.4 ꢀ 6.8
49.9 ꢀ 2.5
57.9 ꢀ 7.2
52.2 ꢀ 3.8
85.4 ꢀ 3.1
49.4 ꢀ 7.1
66.4 ꢀ 7.5
61.0 ꢀ 6.6
44.2 ꢀ 5.1
57.5 ꢀ 6.3
72.4 ꢀ 4.9
56.2 ꢀ 9.9
65.1 ꢀ 7.2
54.9 ꢀ 6.2
61.7 ꢀ 5.0
49.8 ꢀ 5.3
94.4 ꢀ 8.5
78.9 ꢀ 3.6
74.1 ꢀ 5.4a
83.4 ꢀ 5.2
80.9 ꢀ 7.0
93.8 ꢀ 5.2
88.7 ꢀ 6.1
65.4 ꢀ 8.8a
79.1 ꢀ 6.9
74.6 ꢀ 7.5a
71.8 ꢀ 6.7a
78.2 ꢀ 3.5a
66.6 ꢀ 5.9a
59.6 ꢀ 4.7a
85.7 ꢀ 2.8
88.0 ꢀ 9.5
65.1 ꢀ 7.5a
82.1 ꢀ 6.9
66.7 ꢀ 6.9a
79.3 ꢀ 3.0
42.9 ꢀ 5.1a
100.3 ꢀ 3.3
87.7 ꢀ 0.8
80.1 ꢀ 6.0a
88.6 ꢀ 5.3
81.5 ꢀ 2.3
94.8 ꢀ 0.2
83.5 ꢀ 3.7
66.4 ꢀ 6.9a
82.5 ꢀ 8.0
88.0 ꢀ 7.2
76.4 ꢀ 4.8a
81.3 ꢀ 3.3
68.6 ꢀ 7.0a
67.8 ꢀ 3.3a
82.1 ꢀ 1.3
88.7 ꢀ 6.5
55.9 ꢀ 10.6a
83.9 ꢀ 2.3
77.2 ꢀ 6.6a
85.1 ꢀ 3.6
45.9 ꢀ 4.6a
92.3 ꢀ 3.3
85.7 ꢀ 4.4
77.8 ꢀ 5.5a
94.6 ꢀ 7.4
79.9 ꢀ 8.3
78.2 ꢀ 5.5
73.4 ꢀ 2.1a
68.6 ꢀ 6.3a
91.6 ꢀ 5.8
89.9 ꢀ 2.1
82.2 ꢀ 5.2
87.1 ꢀ 2.1
72.4 ꢀ 7.4*
81.9 ꢀ 3.2
75.6 ꢀ 5.1a
90.7 ꢀ 5.4
54.7 ꢀ 7.2a
79.4 ꢀ 7.3
79.6 ꢀ 4.9
78.6 ꢀ 8.3
46.9 ꢀ 5.8a
Control
IIa
IIe
IIIa
V
12.2 ꢀ 0.63
9.5 ꢀ 0.68
10.9 ꢀ 1.03
11.0 ꢀ 0.91
9.1 ꢀ 1.00
9.6 ꢀ 1.11
9.5 ꢀ 0.49
12.9 ꢀ 0.95
9.2 ꢀ 0.83
11.8 ꢀ 0.82
15.1 ꢀ 1.34
13.0 ꢀ 0.85
10.3 ꢀ 1.03
12.0 ꢀ 0.92
9.4 ꢀ 0.51
13.3 ꢀ 1.33
12.6 ꢀ 1.13
9.3 ꢀ 0.94
8.9 ꢀ 0.73
9.4 ꢀ 0.94
13.1 ꢀ 0.78
12.1 ꢀ 0.83
17.5 ꢀ 1.33a
19.8 ꢀ 1.59a
19.1 ꢀ 1.46a
17.0 ꢀ 1.11a
17.1 ꢀ 1.52a
17.0 ꢀ 1.89a
15.0 ꢀ 1.02
10.9 ꢀ 0.56
13.5 ꢀ 0.75
24.9 ꢀ 1.38a
24.1 ꢀ 1.65a
14.6 ꢀ 0.93
14.6 ꢀ 1.24
17.0 ꢀ 0.43a
16.9 ꢀ 1.18
21.0 ꢀ 2.47a
17.2 ꢀ 1.66a
13.0 ꢀ 1.03
17.1 ꢀ 1.66a
17.6 ꢀ 0.32a
12.2 ꢀ 1.18
17.2 ꢀ 1.11a
21.6 ꢀ 1.54a
19.2 ꢀ 1.00a
17.3 ꢀ 1.13a
17.0 ꢀ 1.25a
9.32 ꢀ 0.89
20.5 ꢀ 1.19a
15.3 ꢀ 1.17
11.5 ꢀ 0.85
19.4 ꢀ 0.61a
17.8 ꢀ 2.64a
18.4 ꢀ 1.47a
15.6 ꢀ 1.02
13.6 ꢀ 1.14
18.1 ꢀ 1.36a
18.4 ꢀ 0.69a
18.0 ꢀ 1.23a
16.1 ꢀ 1.20
17.6 ꢀ 1.45a
18.2 ꢀ 0.28a
VIc
VIId
VIIIc
VIIId
XIb
XIc
XIIb
XIIc
XIII
XIV
XVa
XVa
XVIb
XVIIb
XVIIIb
XIXb
Tramadol
XVIb
XVIIb
XVIIIb
XIXb
Indomethacin
a
P < 0.05: Statistically significant from the control using one way ANOVA (Two
Values represent the mean ꢀ S.E. of six animals for each groups.
a
sided Dunnett as Post Hoc test).
P < 0.05: Statistically significant from Control. (Dunnett’s test).
4. Conclusion
XVIb, XVIIIb) that were docked into the same pocket of Indo-
methacin in COX-2 and have shown good docking results and good
fitting into the active site. It has been found that these compounds
have good antiiflammatory effect comparing to Indomethacin. By
the use of molecular modeling we realized the mechanism of their
effects that could be their interactions with the same residues that
interact with Indomethacin.
This study includes the synthesis of three series of novel
derivatives of spiro quinazoline-2,10-cyclohexan-4(1H)-ones
attached to various aromatic and/or heterocyclic ring systems such
as: benzenesulfonamide, pyrazoline, oxazoline, pyrimidin-2-thi-
one, 2-oxo(imino)pyridine, imidazoline and thiazolidinone.
Different nineteen derivatives were evaluated as anti-inflam-
matory and analgesic agents in experimental animals. It has been
found that the derivatives IIe, VIIIc, XIc, XIIb, XIIc, XVIb exhibited
the dual pharmacological activities with superior gastrointestinal
safety profile when compared to indomethacin except VIIIc which
resulted in ulcer lesions in many of the experimental rats.
Surprisingly, compound XIc exhibited no ulcerogenic effect in all of
the experimental animals. Thus, it can be concluded that benze-
nesulfonamide moiety, pyridine and thiazolidinone ring systems
are important for both anti-inflammatory and analgesic activity of
potent safety margin profiles towards G.I.T.
5. Experimental
5.1. Chemistry
All melting points are uncorrected, elemental analyses were
carried out at the Micro-analytical Laboratory, Central Services
Laboratory, National Research Centre, Dokki, Cairo. Infrared spectra
were recorded on FT/IR-330E, Fourier transform, Infrared spec-
trometer at cmꢁ1 scale using KBr discs. 1H NMR spectra were
determined using JEOL EX-270 or JEOL ACA500 NMR spectrometers
We have also designed the top eleven anti-inflammatory qui-
nazoline compounds (IIe, VIId, VIIIc, XIb, XIc, XIIb, XIIc ,XIII, XIV,
and measured in
spectra were measured using mass spectrometer Finnigan MAT
d scale using TMS as an internal standard. Mass
The Anti-inflammatory Effect of the Tested compounds
350
300
250
200
150
100
50
after 4h.
after 3h.
after 2h.
0
Tested Compmounds
Fig. 1.

K.M. Amin et al. / European Journal of Medicinal Chemistry 45 (2010) 2117–2131
2123
The Analgesic Effect of the Tested Compounds
40
35
30
25
20
15
10
5
after 2h.
after 1 .
h
0
I
l
c
I
I
d
I
c
a
e
d
b
I
b
I
I
XV
c
b
c
b
I
XV
Ia
Ib
o
I
V
a
XV
V
I
I
I
r
II
II
II
II
Xb
XI
XI
t
VI
XI
XI
XI
XI
VI
XI
VI
V
n
VI
adol
am
X
Co
r
T
Tested Compounds
Fig. 2.
SSQ-7000 and GCMS-QP 1000EX Shimadzu Gas Chromatography
MS Spectrometer. All reactions were followed up by TLC (aluminum
sheets, Merck plates) using CHCl3/CH3OH (9:1 v/v) eluent and
detected by UV lamp.
ring), 4.31, 6.81, 9.50 10.20 (4s, 2H, 3H, NH2, 3NH, exchangeable
with D2O) and 7.10–8.00 (m, 8H, aromatic-H). MS (m/z): Mþ 413
(5%) and214 (100%).
5.1.2.3. Spiro [(2H,3H)-3(4-(20-methyloxazole) sulfonylphenyl) qui-
nazoline-2,10-cyclohexan]-4(1H) one (IIc). Crystallized from iso-
propanol/pet.ether to give yellowish crystals, mp 230–232 ꢂC, yield
(65%). Analysis for C23H24N4O4S, M.wt.(452.54), Calced.: %C, 61.04;
H, 5.34; N, 12.38; S, 7.08. Found: %C, 61.29; H, 5.22; N, 12.63; S, 7.26.
IR (KBr, cmꢁ1): 3471, 3343 (2NH), 3031(CH-aromatic), 1680 (CO,
5.1.1. Spiro [2H-3,1-benzoxazine-2,10-cyclohexan]-4(1H)-one (I)
This compound was prepared according to a reported method
[39,40]. mp 143 ꢂC.
5.1.2. Spiro [(2H, 3H)-3(4-substituted aminosulfonylphenyl)-
quinazoline-2,10-cyclohexan]-4(1H) ones (IIa–e)
quinazolinone) and 1316 (SO2NH). 1H NMR (DMSO-d6 ,
d ppm): 2.02
General method: a mixture solution of I (2.17 g, 10 mmol) and
(10 mmol) of the appropriate sulfa drug namely, sulfanilamide,
sulfaguanidine, sulfamethoxazole, sulfapyridine and sulfadiazine in
20 mL glacial acetic acid containing (1.64 g, 20 mmol) sodium
acetate anhydrous, was refluxed for 15 h. Upon pouring on crushed
ice/water, white crystals were obtained, filtered, washed with
water and recrystallized from proper solvent.
(s,10H, cyclohexane ring), 2.30 (s, 3H, CH3), 6.82,10.20 (2s, 2H, 2NH,
exchangeable with D2O) and 7.10–8.00(m, 8H, aromatic-H and 1H,
isoxazole ring). MS (m/z): Mþ 452 (4%) and 62 (100%).
5.1.2.4. Spiro [(2H,3H)-3(4-(40pyridinyl)-aminosulfonylphenyl) qui-
nazoline-2,10-cyclohexan]-4(1H) one (IId). Crystallized from iso-
propanol/pet.ether to give white crystals, mp 212–214 ꢂC, yield
(60%). Analysis for C24H24N4O3S, M.wt.(448.55). Calced.: %C, 64.26;
H, 5.39; N, 12.49; S, 7.14. Found: %C, 64.37; H, 5.51; N, 12.62; S, 7.46;
IR (KBr, cmꢁ1): 3355, 3349 (2NH), 3034 (CH-aromatic), 2936 (CH-
alicyclic), 1695 (CO, quinazolinone) and 1327 (SO2NH). MS (m/z):
5.1.2.1. Spiro[(2H,3H)-3(4-aminosulfonylphenyl)quinazoline-2,10-cyc-
lohexan]4(1H)one (IIa). Crystallized from isopropanol to give white
crystals, mp 203–205 ꢂC, yield 60%. Analysis for C19H21N3O3S,
M.wt.(371.47). Calced.: %C, 61.43; H, 5.69; N, 11.31; S, 8.63. Found: %
C, 61.28; H, 5.29; N, 11.93; S, 8.38. IR (KBr, cmꢁ1): 3477, 3470 (NH,
NH2), 3066 (CH-aromatic), 2890 (CH-aliphatic), 1680 (CO,quinazo-
linone) and 1340 (SO2NH). MS (m/z): Mþ 371 (30%) and 119 (100%).
M
þ 448 (10%) and 93 (100%).
5.1.2.5. Spiro [(2H,3H)-3(4-(20-diazine)aminosulfonylphenyl) quina-
zoline-2,10-cyclohexan]-4(1H) one (IIe). Crystallized from iso-
propanol to give brown crystals, mp 203–205 ꢂC, yield (60%).
Analysis for C23H23N5O3S, M.wt.(449.53), Calced.: %,C, 61.45; H,
5.15; N, 15.57; S, 7.13. Found: %C, 61.63; H, 5.48; N, 15.72; S, 7.46. IR
(KBr, cmꢁ1): 3414, 3320 (2NH), 3028 (CH-aromatic), 1680 (CO,
quinazolinone), 1620 (C]N) and 1325 (SO2NH). MS (m/z): Mþ 449
(24%) and 255(100%).
5.1.2.2. Spiro[(2H,3H)-3(4-guanidosulfonylphenyl)quinazoline-2,10-
cyclohexan]-4(1H)one (IIb). Crystallized from ethanol to give
yellowish crystals, mp 256–258 ꢂC, yield; (65%). Analysis for
C20H23N5O3S, M.wt.(413.50). Calced.: %C, 58.09; H, 5.60; N, 16.93; S,
7.75. Found: %C, 57.84; H, 5.81; N, 16.61; S, 7.82. IR (KBr, cmꢁ1):
3467, 3432, 3338, 3211(3NH,NH2), 1681 (CO, quinazolinone) and
1315 (SO2NH). 1H NMR (DMSO-d6 ,
d
ppm): 2.02 (s,10H, cyclohexane
5.1.3. Spiro[(2H,3H)-3(4-substituted sulfonylurea/(thiourea)phenyl)
quinazoline-2,10-cyclohexan]-4(1H) ones (IIIa–c)
General method: a mixture of IIa (1.85 g, 5 mmol) and anhydrous
potassium carbonate (1.38 g, 10 mmol,) in dry acetone (50 mL) was
refluxed with continuous stirring for 1.5 h. While hot, a solution of
the appropriate iso/isothiocynate derivatives namely: ethyl-
isothiocyanate, cyclohexylisocyanate and phenyl isocyanate
(7.5 mmol) in dry acetone was added dropwisely and the reflux was
continued for further 18 h. The excess acetone was removed under
reduced pressure and the obtained solid residue was washed with
water, filtered and recrystallized from the proper solvent.
Table 3
Ulcerogenic effect.
Group
Ulcer index
No. of ulcer
No. Of rats with
ulcer/5
Severity of ulcer
Control (ethanol)
IIe
VIIIc
XIc
XIIb
XIIc
XVIb
Indomethacin
8.2 ꢀ 0.86
0.4 ꢀ 0.40a
7.0 ꢀ 1.64
0.0 ꢀ 0.00a
2.6 ꢀ 1.66a
1.6 ꢀ 1.36a
0.4 ꢀ 0.40a
5.8 ꢀ 1.77
19.4 ꢀ 2.20
0.6 ꢀ 0.60a
10.4 ꢀ 2.86
0.0 ꢀ 0.00a
3.2 ꢀ 1.96a
2.0 ꢀ 1.76a
0.4 ꢀ 0.40a
9.8 ꢀ 3.23a
5
1
5
0
2
2
1
5
5.1.3.1. Spiro
[(2H,3H)-3(4-ethyl-sulfonylthioureaphenyl)quinazo-
line-2,10-cyclohexan]-4(1H) one (IIIa). Crystallized from ethanol to
give yellowish brown crystals, mp170–172 ꢂC, yield (70%). Analysis
for C22H26N4O3S2, M.wt.(458.61). Calced.: %C, 57.61; H, 5.71; N,
12.21; S,13.98. Found: %C, 57.82; H, 5.82; N, 12.52; S,14.05. IR (KBr,
Values represent the mean ꢀ S.E. of five animals for each group.
a
P < 0.05: Statistically significant from ethanol treated rats. (Kruskal Wallis,
followed by Mann Whitney test).

2124
K.M. Amin et al. / European Journal of Medicinal Chemistry 45 (2010) 2117–2131
The Ulcerogenic effect of Most Active Compounds
25
20
15
10
5
No.of Ulcers
Severity of Ulcer
0
Most Active Compounds
Fig. 3.
cmꢁ1): 3340, 3333, 3247 (3NH), 1710 (CO), 1320 (SO2NH) and 1160
(C]S). 1H NMR (CDCL3,
ppm): 1.20 (t, 3H, CH3, ethyl group), 1.81
(s, 10H, cyclohexane ring), 3.62 (q, 2H, CH2, ethyl group), 7.20–8.00
(m, 8H, aromatic-H) and 6.70, 9.25 and 10.20 (3s, 3H, 3NH,
exchangeable with D2O). MS (m/z): Mþ 458 (5%) and 172 (100%).
refluxed for 6 h. The reaction was poured onto ice/water and
neutralized with diluted solution of NaOH. The obtained product
was filtered off, washed with water and recrystallized from ethanol
to give yellowish brown crystals, mp 165–167 ꢂC, yield (65%).
Analysis for C21H22N2O2, M.wt. (334.41). Calced.: %C, 75.42; H, 6.62;
N,8.37. Found,: %C, 75.20; H, 6.84; N, 8.02. IR (KBr, cmꢁ1): 3423
(NH), 3053 (CH, aromatic), 2930 (CH, alicyclic) and 1673,1670
(2CO). 1H NMR (DMSO-d6,d ppm): 2.12 (s,10H, cyclohexane ring),
2.60 (s, 3H, COCH3), 7.20–8.00 (m, 8H, aromatic-H) and 10.20 (s, 1H,
d
5.1.3.2. Spiro [(2H,3H)-3(4-cyclohexylsulfonylureaphenyl) quinazo-
line-2,10-cyclohexan]-4(1H) one. (IIIb). Crystallized from ethanol to
give white crystals, mp 226–228 ꢂC, yield (75%). Analysis for
C26H32N4O4S, M.wt.(496.64). Calced.: %C, 62.88; H, 6.49; N, 11.28; S,
6.45. Found: %C, 63.04; H, 6.40; N, 11.53; S, 6.37. IR (KBr, cmꢁ1):
3320, 3310, 3250 (3NH), 2930 (CH-alicyclic), 1710, 1650 (2CO) and
NH,exchangeable with D2O). 13C NMR (DMSO-d6,
d ppm): 26.32
(CH3), 38.78–40.32 (spiro cyclohexane carbons), 71.8 (spiro head
carbon), 118.04–143.61 (aromatic carbons), 168 (CO,quinazoline
ring), 196.39 (CO, acetyl). MS (m/z): Mþ 334 (14%), and 119 (100%).
1311 (SO2NH). 1H NMR (CDCL3,
d ppm): 1.20–1.11 (m, 6H, cyclo-
hexyl), 1.50–1.50 (m, 4H, cyclohexyl), 2.00(s, 10H, cyclohexane
ring), 4.20 (m, 1H, CH–N), 7.02–7.92 (m, 8H, aromatic-H), 6.80, 9.10,
and10.10 (3s, 3H, 3NH, exchangeable with D2O). MS (m/z): Mþ 496
(2%), [M þ ꢁ 2] 494 (20%) and 56 (100%).
5.1.6. Spiro [(2H, 3H)-3-(substituted acryloylphenyl)quinazoline-
2,10-cyclohexan]-4(1H)-ones (VIa–c)
General method: a mixture of compound V (3.34 g, 10 mmol) and
the appropriate aldehyde namely: benzaldehyde, N,N-dimethyl
aminobenzaldehyde and furan-2-carboxaldehyde (10 mmol) in 5%
ethanolic sodium hydroxide solution (40 mL) was refluxed while
stirring for 15 h. The reaction mixture was poured onto ice/cold
water and neutralized with dil. HCl. The formed precipitate was
filtered off and recrystallized from the proper solvent to give
compounds VIa,b, respectively.
5.1.3.3. Spiro [(2H,3H)-3(4-phenylsulfonylureaphenyl)quinazoline-
2,10-cyclohexan]-4(1H) one (IIIc). Crystallized from ethanol to give
yellowish white crystals, mp 163–165 ꢂC, yield (78%). Analysis for
C26H26N4O4S, M.wt.(490.60). Calced.: %C, 63.65; H, 5.34; N, 11.42; S,
6.53. Found: %C, 63.83; H, 5.28; N, 11.63; S, 6.71. IR (KBr, cmꢁ1):
3340, 3299, 3109 (3NH), 2907 (CH-alicyclic), 1689, 1655 (2CO) and
1312 (SO2NH). MS (m/z):Mþ 490 (4%) and 172 (100%).
5.1.4. Spiro [(2H,3H)-3(4-aminosulfonylphenyl)-1-ethyl-
quinazoline-2,10-cyclohexan]-4(1H) one (IV)
A mixture of compound IIa (1.85 g, 5 mmol,) and iodoethane
(0.78 g, 5 mmol) in 100 mL ethanol containing KOH (0.28 g,
5 mmol) was refluxed for 6 h. The reaction mixture was concen-
trated and acidified with diluted HCl. The separated solid was
filtered off and recrystallized from ethanol to give white crystals,
mp120–122 ꢂC, yield (60%). Analysis for C21H25N3O3S,
M.wt.(399.52). Calced.: %C, 63.13; H, 6.30; N, 10.51; S, 8.02. Found:
C, 63.41; H, 6.51; N, 10.31; S, 8.15. 1H NMR (DMSO-d6,d ppm): 0.90–
1.00 (t, 3H, CH3, ethyl group), 2.11 (s, 10H, cyclohexane ring), 3.00–
3.21 (q, 2H, CH2, ethyl group), 7.00–8.00 (m, 8H, aromatic-H) and
4.55 (s, 2H, NH2 exchangeable with D2O). 13C NMR (DMSO-d6,
d
ppm): 15.2 (CH3), 39.64 (CH2), 39.8–40.00 (spiro cyclohexyl
carbons), 71.8 (spiro head carbon), 119.17–144.23 (aromatic
carbons) and 155 (CO, quinazolinone ring). MS (m/z): Mþ 399 (5%)
and 254 (100%).
5.1.5. Spiro [(2H, 3H)-3-(4-acetylphenyl) quinazoline-2,10-
cyclohexan]ꢁ4(1H)-one (V)
A mixture of the benzoxazine derivatives I (2.17g, 10 mmol,) and
p-aminoacetophenone (1.35 g, 10 mmol) in glacial acetic acid
(20 mL) in the presence of sodium acetate (1.64 g, 20 mmol,) was
Fig. 4. Showing Indomethacin interaction with Tyr 355 –OH group with the –CO group
of Indomethacin carboxylic group with distance 1.03 Ao. Image was taken using
Ligplot.

K.M. Amin et al. / European Journal of Medicinal Chemistry 45 (2010) 2117–2131
2125
Fig. 5. (A) Shows the 3D crystal structure of COX-2 complexed with Indomethacin (pdb: 4COX). (B) Indomethacin (In green color) fitting into the pocket. The structure was viewed
by PDB simpleViewer. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article).
5.1.6.1. Spiro [(2H,3H)-3-(phenylacryloylphenyl) quinazoline2,10-
cyclohexan]-4(1H)-one(VIa). Crystallized from ethanol to give
brown crystals, mp 135–137 ꢂC, yield(60%).. Analysis for
C28H26N2O2, M.wt.(422.52). Calced.: %C, 79.59; H, 6.20; N, 6.63.
Found: %C, 79.38; H, 6.48; N, 6.41. IR (KBr, cmꢁ1): 3330 (NH),
3044(CH, aromatic), 1670, 1650 (2CO). 1H NMR (DMSO-d6,d ppm):
2.04 (s, 10H, cyclohexane ring), 6.68–6.75 (d,d, 2H, CH]CH,
J ¼ 5.6 Hz), 7.55–8.10 (m,13H, aromatic-H) and 10.12 (s, 1H, NH,
exchangeable with D2O). MS (m/z): Mþ 422 (5%) and 120 (100%).
5.1.6.2. Spiro[(2H,3H)-3-(N,N-dimethylaminophenyl–acryloylphenyl)-
quinazoline2,10-cyclo hexan]-4(1H)-one (VIb). Crystallized from
ethanol to give yellow crystals, mp 145 ꢂC, yield (70%). Analysis for
C30H31N3O2, M.wt.(465.59).Calced.: %C, 77.39; H, 6.70; N, 9.02.
Found: %C, 77.34; H, 6.58; N, 9.33. IR (KBr, cmꢁ1): 3325 (NH) and
1685, 1646 (2CO). 1H NMR (DMSO-d6,d ppm): 2.04 (s, 10H, cyclo-
hexane ring), 3.00(s, 6H, N(CH3)2), 6.68–6.75 (d,d, 2H, CH]CH,
J ¼ 5.4 Hz), 7.55–8.10 (m,12H, aromatic-H) and 10.12 (s, 1H, NH,
exchangeable with D2O).MS (m/z): Mþ 465 (5%) and 266 (100%).
A
COOH
S
O
S
5.1.6.3. Spiro [(2H,3H)-3-(20furanoylacryloylphenyl)quinazoline2,10-
cyclohexan]-4(1H)-one (VIc). Crystallized from ethanol to give
yellowish brown crystals, mp 125–127 ꢂC, yield (85%). Analysis for
C26H24N2O3, M.wt.(412.49). Calced.: %C, 75.70; H, 5.86; N, 6.79.
Found: %C, 75.86; H, 5.94; N, 6.55. IR (KBr, cmꢁ1): 3326 (NH),
3044(CH, aromatic), 2995(CH-alicyclic) and 1699, 1641 (2CO). MS
(m/z): Mþ 412 (5%) and 120 (100%).
HO
Tyr 355
N
O
NH2
NH
HOOC
H2N
O
NH
N
H
NH2
Arg 120
N
5.1.7. Spiro [(2H, 3H)-3-(4-(5-aryl)-4,5-dihydro-1H-pyrazol-3-yl-
phenyl) quinazoline2,10 cyclohexan]-4(1H)-ones (VIIa–d)
N
General method: a mixture of the chalcone VIa–b (30 mmol) and
hydrazine hydrate 98% (3 mL, 90 mmol) in absolute ethanol was
B
N
HO
O
N
NH2
O
OH
Ser 530
O
HN
N
S
O
S
NH2
H2N
HN
OH
H
N
NH2
COOH
HOOC
Tyr 355
Arg 120
Fig. 6. A) Possible interactions of compound XVIb with Arg 120 and Tyr 355. B)
Fig. 7. illustrates compound XVIb (in Stick) and how it fits into the pocket. The protein
Possible hydrogen bond formation with –OH of Ser 530.
is presented as cartoon and colored as spectrum.

2126
K.M. Amin et al. / European Journal of Medicinal Chemistry 45 (2010) 2117–2131
Table 4
illustrates the calculated affinities and RMSD for the top 11 anti-inflammatory
compounds and Indomethacin.
Compound
Ranking for biological anti- Affinity Kcal/
RMSD Deviation
from best mode
inflammatory effect
mol
XVIb
XIIc
VIIIc
IIe
XIII
XIc
XVIIIb
XIb
XIIb
VIId
XIV
1
2
3
4
5
6
7
8
ꢁ5.4
ꢁ5.8
ꢁ5.5
ꢁ5.2
ꢁ5.6
ꢁ5.7
ꢁ5.4
ꢁ5.9
ꢁ5.8
ꢁ5.2
ꢁ5.1
ꢁ5.0
4.07
9.5
11.6
15.02
10.3
9.2
8.3
9.0
9.3
9.5
9
10
11
7.2
4.6
Indomethacin –
5.1.7.2. Spiro [(2H, 3H)-3-(4-(5(20furanoyl))-4,5-dihydro-1H-pyr-
azol-3-yl-phenyl)quinazoline 2, 10-cyclohexan]-4(1H)-one (VIIb). -
Crystallized from methanol to give yellowish brown crystals, mp
150–152 ꢂC, yield (60%). Analysis for C26H26N4O2, M.wt.(426.53).
Calced.: %C, 73.21; H, 6.14; N, 13.13; Found: %C, 72.99; H, 6.05; N,
13.42. IR (KBr, cmꢁ1): 3325 (2NH), 2995 (CH, alicyclic), 1700 (CO),
Fig. 8. Distance between –C]O of quinazoline ring and –OH group of XVIb.
and 1600(C]N). 1H NMR (DMSO-d6,
d ppm): 2.00 (s, 10H, cyclo-
refluxed for 3 h. Upon cooling, white crystals were obtained,
filtered off and recrystallized from the proper solvent to obtain the
compounds VIIa,b, respectively. When the reaction was carried out
in glacial acetic acid (20 mL) and refluxed for 4 h, white crystals
were obtained upon pouring onto ice/water. The products were
filtered off and recrystallized from the proper solvents to obtain the
desired derivatives VIIc,d, respectively.
hexane ring), 3.40, 3.80 (d,d, 2H, CH2, pyrazoline ring, J ¼ 5.6 Hz),
6.50 (t,1H, CH, pyrazoline ring), 7.50–8.28 (m,11H, aromatic-H) and
10.21,11.45 (2s, 2NH, exchangeable with D2O). MS (m/z): Mþ 426
(5%) and 279 (100%).
5.1.7.3. Spiro [(2H, 3H)-3-(4-(5-p-N,N-dimethyl amino phenyl)-4,5-
dihydro-1acetyl-pyrazol-3-yl-phenyl)quinazoline 2, 10-cyclohexan]-
4(1H)-one (VIIc). Crystallized from isopropanol to give yellowish
brown crystals, mp 243–245 ꢂC,yield (60%). Analysis for
C32H35N5O2, M.wt.(521.66). Calced.: %C, 73.67; H, 6.76; N, 13.42;
Found: %C, 73.48; H, 6.69; N, 13.64; IR (KBr, cmꢁ1): 3352 (NH), 3046
(CH, aromatic), 2929 (CH, alicyclic), 1700, 1677 (2C0) and
1608(C]N). MS (m/z) :Mþ 521 (10%) and 74 (100%).
5.1.7.1. Spiro [(2H, 3H)-3-(4-(5-p-N,N-dimethylaminophenyl)-4,5-
dihydro-1H-pyrazol-3-yl-phenyl)quinazoline
2,
10-cyclohexan]-
4(1H)-one (VIIa). Crystallized from methanol to give orange crys-
tals, mp 115–117 ꢂC, yield (85%). Analysis for C30H33N5O,
M.wt.(479.62). Calced.: %C, 75.12; H, 6.93; N, 14.60. Found: %C,
75.48; H, 6.85; N, 14.76. IR (KBr, cmꢁ1): 3337, 3211 (2NH), 1690 (CO)
and 1595(C]N). 1H NMR (DMSO-d6,
d ppm): 2.00 (s, 10H, cyclo-
5.1.7.4. Spiro [(2H, 3H)-3-(4-(5(20furanoyl))-4,5-dihydro-1acetyl-
pyrazol-3-yl-phenyl) quinazoline 2, 10-cyclohexan]-4(1H)-one
(VIId). Crystallized from isopropanol to give brown crystals, mp
245–247 ꢂC, yield (65%). Analysis for C28H28N4O3, M.wt.(468.56).
Calced.: %C, 71.77; H, 6.02; N, 11.95. Found: %C, 72.03; H, 6.21; N,
11.74. IR (KBr, cmꢁ1): 3309 (NH), 3042 (CH, aromatic), 2894 (CH,
alicyclic), 1698, 1652 (2CO) and 1560 (C]N). 1H NMR (DMSO-d6,d
ppm): 2.00 (s,10H, cyclohexane ring), 2.20 (s, 3H, COCH3), 2.90, 3.22
hexane ring), 3.00 (s, 6H, N(CH3)2), 3.40, 3.80 (d,d, 2H, CH2, pyr-
azoline ring, J ¼ 5.6 Hz), 6.51 (t, 1H, CH, pyrazoline ring), 7.50–8.28
(m, 12H, aromatic-H) and 10.21,11.45 (2s, 2NH, exchangeable with
D2O). MS (m/z): Mþ 479(4%), and 148 (100%).
Fig. 9. The possible H-bond formation between both Arg 120 NH2 group and Tyr 355
–OH group and two carbonyl groups in XVIb. The protein is shown in catroon view and
colored as salmon while both the compound, Tyr, and Arg are shown as sticks.
Fig. 10. compound IIe measured distance between sulfonyl group and –OH group of
Tyr 385.

K.M. Amin et al. / European Journal of Medicinal Chemistry 45 (2010) 2117–2131
2127
Fig. 11. Hydrogen bond formation between XIV –C]O and –OH group of Tyr 355.
(d,d, 2H, CH2, pyrazoline ring, J ¼ 5.6 Hz), 6.11 (t, 1H, CH of pyr-
azoline ring), 7.12–8.00 (m, 11H, aromatic-H) and 10.22 (s, 1H, NH).
MS (m/z): Mþ 468 (3%) and 78 (100%).
5.1.8. Spiro [(2H, 3H)-3-(4-(5-substituted)-4,5-dihydro-1-methyl/
phenyl-1H-pyrazol-3-yl-phenyl) quinazoline-2,10-cyclohexan]-
4(1H)-ones (VIIIa–d)
General method: a mixture of the chalcone derivatives VIa,b
(3 mmol) and methyl hydrazine(0.14 g, 3 mmol) or phenyl hydra-
zine (0.32 g, 3 mmol) in absolute ethanol (30 mL) was refluxed for
6 h. Upon cooling, the obtained product was filtered off and
recrystallized from isopropanol to obtain the titled derivative
VIIIa–d
Fig. 12. Possible interactions between cyano group of compound XIb and –OH of Tyr 385.
crystals, mp 85–87 ꢂC, yield (60%). Analysis for C32H30N4O2,
M.wt.(502.62). Calced.: %C, 76.46; H, 6.01; N, 11.14. Found: %C,
76.63; H, 6.38; N, 10.92. IR (KBr, cmꢁ1): 3363 (NH), 3037 (CH,
aromatic), 2957 (CH, alicyclic), 1677 (CO) and 1628 (C]N). MS (m/
z): Mþ 502 (10%) and 239 (100%).
5.1.8.1. Spiro [(2H, 3H)-3-(4-(5-N,N-dimethylaminophenyl)-4,5-
dihydro-1-methyl-1H-pyrazol-3-yl-phenyl)quinazoline-2,10-cyclo-
hexan]-4(1H)-one (VIIIa). Crystallized from isopropanol to give
yellowish brown crystals, mp 244–246 ꢂC yield (65%). Analysis for
C31H35N5O, M.wt. (493.65) Calced.: %C, 75.42; H, 7.14; N, 14.18;
Found: %C, 75.33; H, 7.29; N, 14.38. IR (KBr, cmꢁ1): 3385 (NH), 3030
(CH, aromatic), 2930 (CH, alicyclic),1670 (C]O) and 1630(C]N). 1H
5.1.9. Spiro(2H,3H)-3-(4-(5-substituted)-4,5-dihydroisoxazole-3-
yl-phenyl)quinazoline2,10-cyclohexan]-4(1H)-ones (IXa,b)
General method: a mixture of compounds VIa,b (3 mmol) and
hydroxylamine hydrochloride (5 mmol) in sodium hydroxide
solution (0.5 g NaOH in 0.5 mL water) in ethanol (60 mL) was
refluxed for 3 h. The product obtained upon cooling was filtered off,
washed with water and recrystallized from the proper solvents to
obtain the desired compounds IXa,b respectively.
NMR (DMSO-d6, d ppm): 2.00 (s, 10H, cyclohexane ring), 2.31 (s, 3H,
CH3), 3.00 (s, 6H, N(CH3)2), 3.40, 3.80 (d,d, 2H, CH2, pyrazoline ring,
J ¼ 5.6 Hz), 6.50 (t, 1H, CH, pyrazoline ring), 7.50–8.28 (m, 12H,
aromatic-H) and 10.21 (s, NH, exchangeable with D2O). MS
(m/z):Mþ 493 (10%), and 73 (100%).
5.1.8.2. Spiro
[(2H,
3H)-3-(4-(5-[20-furanoyl])-4,5-dihydro-1-
5.1.9.1. Spiro [(2H, 3H)-3-(4-(5-p-N,N-dimethylamino-phenyl)-4,5-
dihydroisoxazole-3-yl-phenyl)quinazoline2,10-cyclohexan]-4(1H)-one
(IXa). Crystallized from methanol to obtain yellowish white crys-
tals, mp 250–252 ꢂC, yield (50%). Analysis for C30H32N4O2, M.wt.
(480.61). Calced.: %C, 74.97; H, 6.71; N, 11.65; Found: %C, 74.63; H,
6.59; N, 11.83. IR (KBr, cmꢁ1): 3351(NH), 3042(CH, aromatic),
2920(CH, alicyclic), 1661(CO) and 1620 (C]N). 1H NMR (DMSO-d6,
methyl)-1H-(pyrazol-3-yl-phenyl)quinazoline-2,10-cyclohexan]-4(1H)-
one (VIIIb). Crystallized from isopropanol to give brown crystals,
mp 240–242 ꢂC, yield (60%). Analysis for C27H28N4O2 , M.wt.
(440.55). Calced.: %C, 73.61; H, 6.40; N, 12.71. Found: %C, 73.72; H,
6.89; N, 12.68. IR (KBr, cmꢁ1): 3367(NH), 2928 (CH, alicyclic), 1677
(CO) and 1604 (C]N). MS (m/z): [Mþ þ 2] 442 (10%) and 73 (100%).
d
ppm) :1.80 (s, 10H, cyclohexane ring), 2.58–2.62 (d, d, 2H, CH2,
5.1.8.3. Spiro
[(2H,
3H)-3-(4-(5-p-N,N-dimethylaminophenyl)-
isoxazoline ring, J ¼ 5.4 Hz), 3.00 (s, 6H, N(CH3)2), 6.44 (t, 1H, CH,
isoxazoline ring), 7.00–8.00 (m,11H, aromatic-H) and 10.2 (s, 1H,
NH, exchangeable with D2O).
4,5-dihydro-1-phenyl-1H-pyrazol-3-yl-phenyl)quinazoline-2,10-cyclo-
hexan]-4(1H)-one (VIIIc). Crystallized from isopropanol to give
brown crystals, mp 160–162 ꢂC, yield (85%). Analysis for C36H37N5O,
M.wt.(555.72). Calced.: %C, 77.80; H, 6.71; N, 12.60. Found: %C,
77.73; H, 6.84; N, 12.52. IR (KBr, cmꢁ1): 3312 (NH), 3017 (CH,
aromatic), 2930 (CH, alicyclic), 1672 (CO) and 1600 (C]N). MS (m/
z): Mþ 555 (4%) and 53 (100%).
5.1.9.2. Spiro [(2H, 3H)-3-(4-(5-[20-furanoyl])-4,5-dihydroisoxazole-
3-yl-phenyl)quinazoline 2,10-cyclohexan]-4(1H)-one (IXb). Crystal-
lized from methanol to give yellow crystals, mp 110–112 ꢂC, yield
(55%). Analysis for C26H25N3O3, M.wt. (427.51). Calced.: %C,
73.04; H, 5.89; N, 9.82. Found: %C, 73.29; H, 5.71; N, 9.69. IR
(KBr, cmꢁ1): 3350 (NH), 3040 (CH, aromatic), 2934 (CH, alicy-
clic), 1670 (CO), 1610 (C]N). MS (m/z): Mþ 427 (15%) and 99
(100%).
5.1.8.4. Spiro
[(2H,
3H)-3-(4-5-[20-furanoyl])-4,5-dihydro-1-
phenyl-1H-pyrazol-3-yl-phenylquinazoline-2,10-cyclohexan]-4(1H)-
one (VIIId). Crystallized from isopropanol to give dark brown

2128
K.M. Amin et al. / European Journal of Medicinal Chemistry 45 (2010) 2117–2131
5.1.10. Spiro [(2H, 3H)-3-(4-(6-substituted)-1,2-dihydro-2-
thiopyrimidin-4-yl-phenyl)-quinazoline2,10-cyclohexan]-4(1H)-
ones(Xa,b)
(C^N), 1666, 1650 (2CO) and 1557(C]N). MS (m/z): [Mþþ2] 504
(24%), and 62 (100%).
General method: a solution of the chalcone derivatives VIa,b
(3 mmol), thiourea (0.23 g, 3 mmol) and sodium hydroxide (0.1 g)
in absolute ethanol (30 mL) was refluxed for 6 h. The reaction
mixture was concentrated under vacuum, cooled and neutralized
with dilute HCl. The formed product was filtered off and washed
with water to yield the Xa,b, respectively.
5.1.12. Spiro (2H, 3H)-3-(4-(4-aryl)-1,2-dihydro-3-cyano-2-imino-
pyridin-6-yl-phenyl)- quinazoline-2,10-cyclohexan]-4(1H)-ones
(XIIa–c)
General method: a mixture of the acetyl derivative V (3.34 g;
10 mmol), malononitrile (0.66 g; 10 mmol) and the appropriate
aldehyde namely: benzaldehyde, p-florobenzaldehyde and 2-
hydroxybenzaldehyde (10 mmol) and ammonium acetate (6.16 g;
80 mmol) in n-butanol (50 mL) was heated under reflux for 6 h. The
solid formed upon concentrating the solvent was filtered off,
washed with water and recrystallized from the proper solvent to
obtain the desired product XIIa–c, respectively.
5.1.10.1. Spiro
[(2H,3H)–3-(4-(6-N,N–dimethylaminophenyl)-1,2-
dihydro-2-thiopyrimidin-4-yl-phenyl)quinazoline2,10-cyclohexan]-
4(1H)-one (Xa). Crystallized from isopropanol to give reddish
brown crystals, mp 175–177 ꢂC, yield (71%). Analysis for
C31H31N5OS, M.wt.(521.68). Calced.: %C, 71.37; H, 5.98; N, 13.42; S,
6.14. Found: %C, 71.58; H, 6.77; N, 13.23; S, 6.35. IR (KBr, cmꢁ1):
3384, 3351(2NH), 3043 (CH, aromatic), 2924 (CH, alicyclic), 1660
(CO), 1596 (C]N) and 1227 (C]S). MS (m/z): Mþ 521(10%) and
147(100%).
5.1.12.1. Spiro[(2H,3H)-3-(4-(4-phenyl)-1,2-dihydro-3-cyano-2-imino-
pyridin-6-yl-phenyl) quinazoline-2,10-cyclohexan]-4(1H)-one (XIIa).
Crystallized from glacial acetic acid to give yellow crystals, mp 139–
141 ꢂC, yield (50%). Analysis for C31H27N5O, M.wt. (485.58). Calced.:
%C, 76.67; H, 5.60; N, 14.42. Found: %C, 76.49; H, 5.48; N, 14.64. IR
(KBr, cmꢁ1): 3445, 3337, 3218 (3NH), 3061 (CH, aromatic), 2936
(CH, alicyclic), 2196 (C^N), 1670 (CO) and 1624 (C]N). MS (m/z):
5.1.10.2. Spiro
[(2H,
3H)-3-(4-(6-[20-furanoyl])-1,2-dihydro-2-
thiopyrimidin-4-yl-phenyl)-quinazoline2,10-cyclohexan]-4(1H)-one
(Xb). Crystallized from methanol to give orange crystals, mp 130–
132 ꢂC, yield (50%). Analysis for C27H24N4O2S, M.wt. (468.59).
Calced.: %C, 69.20; H, 5.16; N, 11.95; S, 6.84. Found: %C, 69.53; H,
5.32; N, 12.11; S, 6.77. IR (KBr, cmꢁ1): 3338, 3214 (2NH), 3067 (CH,
aromatic), 2918 (CH, alicyclic), 1660 (CO), 1590 (C]N) and 1273
(C]S). MS (m/z): Mþ 468 (15%) and 112 (100%).
M
þ 486 (18%) and 190 (100%).
5.1.12.2. Spiro[(2H,3H)-3-(4-(4-p-florophenyl)-1,2-dihydro-3-cyano-
2-imino-pyridin-6-yl-phenyl)
quinazoline-2,10-cyclohexan]-4(1H)-
one (XIIb). Crystallized from methanol to give orange crystals, mp
132–134 ꢂC, yield (53%). Analysis for C31H26FN5O, M.wt.(503.58).
Calced.: %C, 73.93; H, 5.20; N, 13.90. Found: %C, 74.15; H, 5.43; N,
13.73. IR (KBr, cmꢁ1): 3438, 3346, 3237 (3NH), 3087 (CH, aromatic),
2928 (CH, alicyclic), 2204 (C^N), 1670 (CO) and 1612 (C]N). MS
(m/z): Mþ 503 (14%) and 62 (100%).
5.1.11. Spiro (2H, 3H)-3-(4-(4-aryl)-1, 2-dihydro-3-cyano-2-oxo-
pyridin-6-yl-phenyl)-quinazoline-2,10-cyclohexan]-4(1H)-ones
(XIa–c)
General method: a mixture of the acetyl derivative V (3.34 g,
10 mmol), ethyl cyanoacetate (1.13 g,10 mmol), and the appropriate
aldehyde namely: benzaldehyde, p-florobenzaldehyde and 2-
hydroxybenzaldehyde (10 mmol) and ammonium acetate (6.16 g,
80 mmol) in n-butanol (50 mL) was heated under reflux for 6 h. The
solid separated upon cooling was filtered off, washed with water
and recrystallized from the proper solvent to obtain the desired
product XIa–c, respectively.
5.1.12.3. Spiro[(2H,3H)-3-(4-(4-hydroxyphenyl)-1,2-dihydro-3-cyano-
2-imino-pyridin-6-yl-phenyl)quinazoline-2,10-cyclohexan]-4(1H)-
one (XIIc). Crystallized from methanol to give light brown crystals,
mp 218–220 ꢂC, yield (57%). Analysis for C31H27N5O2, M.wt.
(501.58). Calced.: %C, 74.23; H, 5.42; N, 13.96. Found: %C, 74.49; H,
5.33; N, 13.68. IR (KBr, cmꢁ1): 3438, 3346, 3237 (3NH), 3087 (CH,
aromatic), 2928 (CH, alicyclic), 2204 (C^N), 1670 (CO) and 1612
(C]N). MS (m/z) :Mþ 502 (8%) and 69 (100%).
5.1.11.1. Spiro[(2H,3H)–3-(4-(4-phenyl)-1,2–dihydro-3-cyano-2-oxo-
pyridin-6-yl-phenyl)-quinazoline-2,10-cyclohexan]-4(1H)-one (XIa).
Crystallized from glacial acetic acid to give bright yellow crystals,
mp over 300 ꢂC, yield (57%). Analysis for C31H26N4O2,
M.wt.(486.58). Calced.: %C, 76.52; H, 5.38; N, 11.51. Found: %C,
76.69; H, 5.56; N, 11.49. IR (KBr, cmꢁ1): 3331, 3237 (2NH), 3079
(CH,aromatic), 2965 (CH,alicyclic), 2128 (C^N), 1660, 1656 (2CO)
and 1601(C]N). MS (m/z): Mþ 486 (16%) and 59 (100%).
5.1.13. Ethyl-4-[spiro[(2H,3H)quinazoline-2,10-cyclohexan]-
4-(1H)-one-3-yl]-benzoate (XIII)
A mixture of the benzoxazine derivative I (2.17 g; 10 mmol) and
ethyl-p-aminobenzoate (1.65 g; 10 mmol) in glacial acetic acid
(30 mL) containing sodium acetate anhydrous (1.64 g; 20 mmol)
was refluxed for 8 h. Upon pouring on ice/water and neutralization
with dilute NaOH solution, white precipitate was obtained which
was filtered off, washed with H2O and recrystallized with iso-
propanol to obtain the desired ester derivative XIII. mp 70–72 ꢂC,
yield (75%). Analysis for C22H24N2O3, M.wt. (364.44). Calced.: %C,
72.50; H, 6.63; N, 7.68. Found: %C, 72.40; H, 6.70; N, 7.60. IR (KBr,
cmꢁ1): 3291 (NH), 3066(CH, aromatic), 2990 (CH, alicyclic) and
5.1.11.2. Spiro[(2H,3H)-3-(4-(4-p-florophenyl)-1,2-dihydro-3-cyano-
2-oxo-pyridin-6-yl-phenyl)quinazoline-2,10-cyclohexan]-4(1H)-one
(XIb). Crystallized from methanol to give yellowish white crystals,
mp over 300 ꢂC, yield (50%). Analysis for C31H25FN4O2, M.wt.
(504.57). Calced.: %C, 73.79; H, 4.99; N, 11.10. Found: %C, 73.82; H,
4.72; N, 10.92. IR (KBr, cmꢁ1): 3339, 3263 (2NH), 3065 (CH,ar-
omatic), 2927 (CH,alicyclic), 2213(C^N), 1665,1656 (2CO) and 1599
(C]N). MS (m/z) :Mþ 504 (24%) and 307 (100%).
1690, 1685 (2CO). 1H NMR (DMSO-d6,
d ppm): 1.21(t, 3H, CH3, ethyl
group), 2.09 (s, 10H, cyclohexane ring), 4.20 (q, 2H, CH2, ethyl
group) and 7.11–8.02 (m, 8H, aromatic-H) and 10.21 (s, 1H, NH,
exchangeable with D2O). MS (m/z): Mþ 364 (10%) and 120 (100%).
5.1.11.3. Spiro[(2H,3H)-3-(4-(4-O-hydroxyphenyl)-1,2-dihydro-3-
cyano-2-oxo-pyridin-6-yl-phenyl)quinazoline-2,10-cyclohexan]-
4(1H)-one (XIc). Cystallized from methanol to give orange crystals,
mp over 300 ꢂC, yield (60%). Analysis for C31H26N4O3,
M.wt.(502.58). Calced.: %C, 74.08; H, 5.21; N, 11.14. Found: %C,
73.99; H, 5.32; N, 11.29. IR (KBr, cmꢁ1): 3331, 3213 (2NH), 2248
5.1.14. 4-[Spiro[(2H,3H)quinazoline-2,10-cyclohexan]-4-(1H)-one-
3-yl]-benzoic acid hydrazide (XIV)
A solution of the ester derivative XIII (3.64 g; 10 mmol) and
hydrazine hydrate 98% (1.6 g; 50 mmol) in absolute ethanol (20 mL)
was refluxed for 6 h upon cooling, the formed precipitate was
filtered off and recrystallized from ethanol to give the hydrazide

K.M. Amin et al. / European Journal of Medicinal Chemistry 45 (2010) 2117–2131
2129
derivative XIV. mp 300–302 ꢂC, yield (67%). Analysis for
C20H22N4O2, M.wt. (350.42). Calced.: %C, 68.55; H, 6.32; N, 15.98.
Found: %C, 68.70; H, 6.13; N, 15.90. IR (KBr, cmꢁ1): 3311, 3219 (NH,
NH2), 3046 (CH, aromatic), 2931(CH, alicyclic), 1672 (CO, quinazo-
line ring) and 1640 (CO, amide). 1H NMR (DMSO-d6,d ppm): 2.00 (s,
10H, cyclohexane ring), 4.32 (s, 2H, NH2, exchangeable with D2O),
7.40–8.02 (m, 8H, aromatic-H) and 9.50, 10.00 (2s, 2H, 2NH,ex-
changeable with D2O).
Crystallized from ethanol to give brownish red crystals, mp 285–
287 ꢂC, yield (65%). Analysis for C27H26N4O3S2, M.wt. (518.67).
Calced.: %C, 62.52; H, 5.05; N, 10.80; S, 12.36. Found: %C, 62.31; H,
5.26; N, 11.00; S, 12.20. IR (KBr, cmꢁ1): 3354, 3209 (2NH), 3052 (CH,
aromatic), 2923(CH, alicyclic), 1700, 1684 (2CO, cyclic amides), 1657
(CONH) and 1603 (C]N). 1H NMR (DMSO-d6,
d ppm): 2.1 (s, 10H,
cyclohexane ring), 3.71 (s, 2H, CH2, thiazolidinone ring,), 6.11 (s, 1H,
CH, thiazolidinone ring), 7.20–8.50 (m, 11H, aromatic-H) and 10.2,
11.5 (2s, 2H, 2NH, exchangeable with D2O). MS (m/z): [M þ ꢁ 2] 516
(8%) and 55 (100%).
5.1.15. 4-[Spiro [(2H, 3H) quinazoline-2,10-cyclohexan]-4-(1H)-
one-3-yl]-benzoic acid [1-substituted aryl meth-(e)-ylidene]-
hydrazides (XVa,b)
Generalmethod: a mixture of compound XIV (3.50 g; 10 mmol)
and the appropriate aldehyde, namely: p-methoxybenzaldehyde
and 2-thiophenecarboxaldehyde (10 mmol) in absolute ethanol
(30 mL) in the presence of few drops of acetic acid, was refluxed for
12 h. The reaction mixture was cooled and the formed precipitate
was filtered off and recrystallized from the proper solvent to obtain
the desired Schiff,s bases XVa,b.
5.1.17. 4-[Spiro[(2H,3H)quinazoline-2,10-cyclohexan]-4-(1H)-one-
3-yl]-N-substituted amido (thioamido) benzoic acid hydrazides
(XVIIa–c)
General method: a mixture of compound XIV (3.50 g; 10 mmol),
the appropriate iso/isothiocyanate, namely: cyclohexyl isocyanate,
cyclohexyl isothiocyanate and/or phenyl isothiocyanate (10 mmol)
and few drops of triethylamine in dry benzene (20 ml) was refluxed
for 12 h. The solvent was evaporated under reduced pressure and
the obtained solid was recrystallized from the proper solvent to
give the compounds XVIIa–c, respectively.
5.1.15.1. 4-[Spiro[(2H,3H)quinazoline-2,10-cyclohexan]-4-(1H)-one-
3-yl]–benzoic acid-[1-(4-methoxy phenyl) meth-(e)-ylidene]-hydra-
zide (XVa). Crystallized from ethanol to give light brown crystals,
mp 241–243 ꢂC, yield (73%). Analysis for C28H28N4O3, M.wt.
(468.66). Calced.: %C, 71.77; H, 6.02; N, 11.95. Found: 71.90; H, 6.30;
N, 11.66. IR (KBr, cmꢁ1): 3315, 3296 (2NH), 1670, 1645 (2CO) and
1604 (C]N). 1H NMR (DMSO-d6,d ppm): 1.89 (s, 10H, cyclohexane
ring), 3.80 (s, 3H, OCH3), 6.00(s, 1H, CH]N), 6.8–7.6(m,12H,
aromatic-H) and 8.10, 10.20 (2s, 2H, 2NH, exchangeable with D2O).
5.1.17.1. 4-[Spiro[(2H,3H)quinazoline-2,10-cyclohexan]-4-(1H)-one-3-
yl]-N-(cyclohexyl) amido benzoic acid hydrazide (XVIIa). Crystal-
lized from isopropanol to give white crystals, mp 146–148 ꢂC, yield
(80%). Analysis for C27H33N5O3, M.wt. (475.59). Calced.: %C, 68.18;
H, 6.99; N, 14.72. Found: %C, 68.22; H, 6.70; N, 14.40. IR (KBr, cmꢁ1):
3392, 3328, 3294,3107(4NH), 3067 (CH-aromatic), 2931 (CH,
alicyclic), 1679, 1660, 1657 (3CO). 1H NMR (DMSO-d6,
d ppm):
1.00–1.20 (m, 6H, cyclohexyl),1.50–1.71 (m, 4H, cyclohexyl), 2.00 (s,
10H, cyclohexane ring), 3.84 (m, 1H, CH–N), 7.61–8.10 (m, 8H,
aromatic-H), 4.21, 7.00, 9.95 and 10.12 (4s, 4H, 4NH, exchangeable
with D2O).
5.1.15.2. 4-[Spiro[(2H,3H)quinazoline-2,10-cyclohexan]-4-(1H)-one-3-
yl]benzoicacid-[1-(2-thienyl)meth-(e)-ylidene]-hydrazide(XVb). Crys-
tallized from ethanol to give reddish brown crystals, mp 240–
242 ꢂC, yield (79%). Analysis for C25H24N4O2S, M.wt. (444.57).
Calced.: %C, 67.54; H, 5.44; N, 12.60; S, 7.21. Found, %C, 67.35; H,
5.60; N, 12.32; S, 6.93. IR (KBr, cmꢁ1): 3346, 3246 (2NH), 1669, 1660
(2CO) and 1592 (C]N). MS (m/z): [Mþ þ1] 445 (13%) and 120
(100%).
5.1.17.2. 4-[Spiro[(2H,3H)quinazoline-2,10-cyclohexan]-4-(1H)-one-3-
yl]-N-(cyclohexyl) thioamido benzoic acid hydrazide (XVIIb). Crys-
tallized from isopropanol to give white crystals, mp 211–213 ꢂC,
yield (73%). Analysis for C27H33N5O2S, M.wt. (491.66). Calced.: %C,
65.96; H, 6.76; N, 14.24; S, 6.52. Found: %C, 65.73; H, 6.98; N, 13.98;
S, 6.76. IR (KBr, cmꢁ1): 3390, 3380, 3370, 3281 (4NH), 3012 (CH-
aromatic), 2929 (CH, alicyclic), 1687, 1647 (2CO) and 1180 (C]S). 1H
5.1.16. 4-[Spiro[(2H,3H)quinazoline-2,10-cyclohexan]-4-(1H)-one-
3-yl]-N-[4-oxo-2-(substituted aryl) thiazolidin-3-yl] benzamides
(XVIa,b)
NMR (DMSO-d6,
d ppm): 1.00–1.11 (m, 6H, cyclohexyl), 1.50–1.71
General method: a mixture of compounds (XVIa,b) (5 mmol) and
thioglycolic acid (0.35 ml; 5 mmol) in dry benzene (20 mL) was
refluxed for 16 h. The excess solvent was evaporated under reduced
pressure and the obtained residue was treated with petroleum
ether (60–80). The solid product was filtered off and washed with
petroleum ether (60–80) to obtain the desired products XVIa,b
respectively.
(m, 4H, cyclohexyl), 2.00(s, 10H, cyclohexane ring), 4.00 (m, 1H,
CH–N), 7.02–7.85 (m, 8H, aromatic-H), 4.25, 9.10, 10.11 and 10.20
(4s, 4H, 4NH, exchangeable with D2O). 13C NMR (DMSO-d6,
d ppm):
32–40 (10 CH2, cyclohexyl and cyclohexane rings), 53.9 (CH–NH),
71.8 (spiro head C), 118–143 (aromatic carbons), 165.8 (CO–NH),
170.0 (CO, quinazoline ring) and 185.0 (C]S).
5.1.17.3. 4-[Spiro[(2H,3H)quinazoline-2,10-cyclohexan]-4-(1H)-one-
3-yl]-N-phenylthioamido benzoic acid hydrazide (XVIIc). Crystal-
lized from ethanol to give light brown crystals, mp192–201 ꢂC, yield
(80%). Analysis for C27H27N5O2S, M.wt.(485.61). Calced.: %C, 66.79;
H, 5.60; N, 14.42; S, 6.60. Found: %C, 66.99; H, 5.78; N, 14.38; S, 6.75.
IR (KBr, cmꢁ1): 3388, 3370, 3339, 3787 (4NH), 3041 (CH, aromatic),
1668 (CO, quinazoline ring), 1632 (CONH) and 1188 (C]S). 1H NMR
(DMSO-d6,d ppm): 2.00 (s, 10H, cyclohexane ring), 7.00–8.00 (m,
13H, aromatic-H) and 9.6, 9.7, 10.2, 10.4 (4s, 4H, 4NH, exchangeable
with D2O). MS (m/z): 485 (10%) and 162 (100%).
5.1.16.1. 4-[Spiro [(2H,3H)- quinazoline-2,10- cyclohexan]-4-(1H)-
one-3-yl]-N-[4-oxo-2-(4-methoxyphenyl) thiazolidin-3-yl] benza-
mide (XVIa). Crystallized from isopropanol to give brown crystals,
mp 95–101 ꢂC, yield (70%). Analysis for C30H30N4O4S, M.wt.
(542.67). Calced.: %C, 66.39; H, 5.57; N, 10.32; S, 5.90. Found: %C,
66.56; H, 5.32; N, 10.10; S, 6.10. IR (KBr, cmꢁ1): 3297, 3272 (2NH),
3041(CH, aromatic), 2968 (CH, alicyclic), 1684, 1680 (2CO, cyclic
amide), 1655 (CONH) and 1601(C]N). 1H NMR (DMSO-d6,
d ppm):
2.1 (s, 10H, cyclohexane ring), 3.70 (s, 2H, CH2, thiazolidinone ring),
3.85 (s, 3H, OCH3), 5.90 (s, 1H, CH of thiazolidinone ring), 7.00–8.50
(m, 12H, aromatic-H) and 10.21,10.50 (2s, 2H, 2NH, exchangeable
with D2O).
5.1.18. 4-[Spiro[(2H,3H)quinazoline-2,10-cyclohexan]-4-(1H)-one-
3-yl]-N-[(2,4-dioxo/2-thio-4-oxo)-1-substituted-imidazolidin-3-
yl]benzamides (XVIIIa–c)
General method: a mixture of compound XVIIa–c (2 mmol),
monochloroacetic acid (0.3 g; 3 mmol) and few drops of pyridine
5.1.16.2. 4-[Spiro[(2H,3H)-quinazoline-2,10-cyclohexan]-4-(1H)-one-3-
yl]-N-[4-oxo-2-(thienyl)
thiazolidin-3-yl]
benzamide
(XVIb).

2130
K.M. Amin et al. / European Journal of Medicinal Chemistry 45 (2010) 2117–2131
were fused for 6 h. The obtained materiel was triturated with
crushed ice, filtered off and crystallized from the proper solvent to
obtain the imidazoline derivatives XVIIIa–c, respectively.
5.1.19.2. 4-[Spiro[(2H,3H)quinazoline-2,10-cyclohexan]-4-(1H)-one-3-
yl]-N-(2,4,5,6-tetrahydro- 2-thioxo-4,6-dioxo-3cyclohexyl pyrimidin-
1-yl) benzamide (XIXb). Crystallized from isopropanol to give white
crystals, mp 128–130 ꢂC, yield (60%). Analysis for C30H33N5O4S,
M.wt. (559.69). Calced.: %C, 64.38; H, 5.94; N, 12.51; S, 5.72. Found,
%C, 64.50; H, 5.73; N,12.85; S, 5.45. IR (KBr, cmꢁ1): 3268, 3110 (2NH),
3055 (CH, aromatic), 2930 (CH, alicyclic), 1695–1670 (3CO lactamic),
5.1.18.1. 4-[Spiro[(2H,3H)quinazoline-2,10-cyclohexan]-4-(1H)-one-3yl]-
N-(2,4-dioxo-1-cyclohexylimidazolidin-3-yl)benzamide (XVIIIa). Cry-
stallized from DMF/H2O to give yellow crystals, mp129–131 ꢂC,
yield (65%). Analysis for C29H33N5O4 , M.wt. (515.61). Calced.: %C,
67.55; H, 6.45; N, 13.58. Found: %C, 67.32; H, 6.44; N, 13.80. IR
(KBr, cmꢁ1): 3349, 3340 (2NH), 2934 (CH, alicyclic), 1675, 1660
(2CO, lactamic). and 1640 (CO, amide). 1H NMR (DMSO-d6,d ppm):
1.10–1.21 (m, 6H, cyclohexyl), 1.50–1.61 (m, 4H, cyclohexyl), 2.00
(s, 10H, cyclohexane ring), 3.91 (m, 1H, CH-N), 4.22 (s, 2H, CH2,
imidazolidine ring), 7.6–8.6 (m, 8H, aromatic-H) and 8.7–9.0 (2S,
2H, 2NH, exchangeable with D2O). MS (m/z):Mþ 516 (5%) and
53(100%).
1640 (CO, amide) and 1182 (C]S). 1H NMR (DMSO-d6,
d ppm): 1.00–
1.21 (m, 6H, cyclohexyl), 1.51–1.81 (m, 4H, cyclohexyl), 2.00 (s, 10H,
cyclohexane ring), 3.60 (s, 2H, CH2), 4.11 (m, 1H, CH–N), 7.20–8.20
(m, 8H, aromatic-H) and 9.11, 10.20 (2s, 2H, 2NH, exchangeable
with D2O).
5.1.19.3. 4-[Spiro[(2H,3H)quinazoline-2,10-cyclohexan]-4-(1H)-one-
3-yl]-N-(2,4,5,6-tetrahydro-2-thioxo-4,6-dioxo-3-phenyl pyrimidin-
1-yl) benzamide (XIXc). Crystallized from ethanol to give brownish
yellow crystals, mp 112–114 ꢂC, yield (73%). Analysis for
C30H27N5O4S, M.wt. (553.64). Calced.: %C, 65.08; H, 4.91; N,12.64; S,
5.79. Found: %C, 65.38; H, 4.77; N, 12.87; S, 5.88. IR (KBr, cmꢁ1):
3214, 3115 (2NH), 3046 (CH, aromatic), 2982 (CH, alicyclic), 1695-
1670 (3CO, lactamic), 1640 (CO, amide) and 1181 (C]S). MS (m/z):
[Mþ þ1] 554 (15%), 73(100%).
5.1.18.2. 4-[Spiro[(2H,3H)quinazoline-2,10-cyclohexan]-4-(1H)-one-
3-yl]-N-(2-thio-4-oxo-1-cyclohexylimidazolidin-3-yl)benzamide
(XVIIIb). Crystallized from DMF/H2O to give white crystals, mp
165–167 ꢂC, yield (65%). Analysis for C29H33N5O3S, M.wt. (531.68).
Calced.: %C, 65.51; H, 6.25; N, 13.17; S, 6.03. Found: %C, 65.26; H,
5.98; N, 13.00; S, 6.25.IR (KBr, cmꢁ1): 3259, 3247 (2NH), 3098(CH,
aromatic), 2929(CH, alicyclic), 1675, 1672 (2CO, lactamic), 1641 (CO,
5.2. Biological screening
amide) and 1181(C]S). 1H NMR (DMSO-d6,
d ppm): 1.10–1.21 (m,
6H, cyclohexyl), 1.50–1.62 (m, 4H, cyclohexyl), 2.00 (s, 10H, cyclo-
hexane ring), 4.11 (m, 1H, CH–N), 4.17 (s, 2H, CH2, imidazolidine
ring), 7.69–8.54 (m, 8H, aromatic-H) and 8.90–9.00 (2S, 2H, 2NH,
exchangeable with D2O). MS (m/z):[Mþ þ 1] 532 (5%) and 53
(100%).
5.2.1. Materials and methods
Animals-Adult rats of both sexes weighing 150–200 g and adult
mice weighing 20–25 g were used in the experiments. Animals
were housed under standardized conditions for light and temper-
ature and received standard rat chow and tap water and libitum.
Animals were randomly assigned to different experimental groups,
each kept in a separate cage. All animal procedures were performed
after approval from the Ethics committee of the National Research
Center and in accordance with the recommendations for the proper
care and use of laboratory animals (NIH publication No.85-23,
revised 1985).
5.1.18.3. 4-[Spiro[(2H,3H)quinazoline-2,10-cyclohexan]-4-(1H)-one-3-
yl]-N-(2-thio-4-oxo-1-phenylimidazolidin-3-yl) benzamide (XVIIIc).
Crystallized from DMF/H2O to give brown crystals, mp 195–
197 ꢂC, yield (65%). Analysis for C29H27N5O3S, M.wt. (525.63).
Calced.: %C, 66.26; H, 5.17; N, 13.32; S, 6.10. Found: %C, 66.54; H,
5.36; N, 13.00; S, 6.40. IR (KBr, cmꢁ1): 3360, 3281 (2NH), 3049
(CH, aromatic), 2987 (CH, alicyclic), 1670, 1665 (2CO, lactamic),
5.2.2. Drugs and chemicals
1645 (CO, amide) and 1183 (C]S). 1H NMR (DMSO-d6,
d
ppm):
Carrageenan Iambda Sigma–Aldrich chemical company (USA),
indomethacin Khahira Pharmaceutical and Chemical Company
(Cairo, Egypt) and tramadol October Pharma (Cairo, Egypt).
2.00 (s, 10H, cyclohexane ring), 4.21 (s, 2H, CH2 of imidazolidine
ring), 7.51–8.20 (m, 13H, aromatic-H-) and 10.11, 11.13 (2S, 2H,
2NH, exchangeable with D2O).
5.2.3. Antiinflammatory testing
5.1.19. 4-[Spiro[(2H,3H)quinazoline-2,10-cyclohexan]-4-(1H)-one-
3-yl]-N-[2,4,5,6-tetrahydro-(2,4,6-trioxo/2-thio-4,6-dioxo)-3-
substituted pyrimidin-1-yl]benzamide (XIXa–c)
General method: a mixture of compound XVIIa–c (2 mmol) and
malonic acid (0.2 g; 2 mmol) in absolute ethanol (20 mL) was
refluxed for 16h. The precipitated solid formed upon cooling, was
filtered and recrystallized from the proper solvent to obtain the
desired derivatives XIXa–c, respectively.
The carrageenan rat paw oedema model of inflammation was
used to evaluate the anti-inflammatory properties of the tested
compounds. Rats were randomly assigned to treatment groups and
sterile carrageenan lambda (100 ul of a 1% solution in saline) was
injected sub-planter into right hind paw of the rat. Carrageenan
caused visible redness and pronounced swelling that was well
developed by 4 h and persisted for more than 48 h. Right hind paw was
measured with a planimeter [53,54] before, and at 1,2,3 and 4 h after
carrageenan injection. Due to water insolubility of the tested
compounds, they were dissolved in DMSO then injected i.p (9 mg/
kg b wt) [37].The control animals were injected (i.p) with
appropriate volume of DMSO. The standard drug was indo-
methacin (10 mg/kg b wt). Different compounds or indometh-
acin were given 1hr before carrageenan injection.
5.1.19.1. 4-[Spiro [(2H, 3H)quinazoline-2,10-cyclohexan]-4-(1H)-one-
3-yl]-N-(2,4,5,6-tetrahydro-2,4,6-trioxo- 3-cyclohexyl pyrimidin-1-
yl) benzamide (XIXa). Crystallized from isopropanol to give white
crystals, mp 235–237 ꢂC, yield (70%). Analysis for C30H33N5O5,
M.wt. (543.62). Calced.: %C, 66.28; H, 6.11; N, 12.88.Found: %C,
65.95; H, 6.38; N, 13.03. IR (KBr, cmꢁ1): 3319, 3251 (2NH), 3016
(CH, aromatic), 2930 (CH, alicyclic), 1694–1670 (4CO, lactamic)
and 1641 (CO, amide). 1H NMR (DMSO-d6,dppm):1.00–1.21
(m, 6H, cyclohexyl), 1.50–1.71 (m, 4H, cyclohexyl), 2.00
(s,10H, cyclohexane ring), 3.62 (s, 2H, CH2), 3.82 (m, 1H, CH–N),
7.61–8.11 (m, 8H, aromatic-H) and 9.99,10.11 (2s, 2H, 2NH,
exchangeable with D2O). MS (m/z): [M þ ꢁ 1] 542 (5%) and 120
(100%).
5.2.4. Analgesia testing
The hot-plate test was performed on mice by using an elec-
tronically controlled hot-plate (ugo Basile, Italy) heated to 52 ꢂC
(ꢀ0.1 ꢂC), for possible centrally mediated analgesic effect of the
drugs.
Nineteen groups of rats each were given vehicle and/or the
different compounds and the last group received tramadol (20 mg/

K.M. Amin et al. / European Journal of Medicinal Chemistry 45 (2010) 2117–2131
2131
[15] M. Kenji, U. Junko, S. Takashi, I. Michio, A.G. Neil, Y. Jin-Chen, T. Shusuke,
O. Shoji, N. Yuji, J. Med. Chem. 46 (2003) 4910–4925.
[16] Archana, V.K. Srivastava, C. Ramesh, K. Ashok, Indian J. Chem. 41B (2002)
2371–2375.
Kg b wt) 60 min prior to testing. Latency to lick a hind paw or
jumping [55] was recorded sequentially before and at 1, 2 h post
treatment.
[17] R. Joachim, P.E. William, O. Stephen, D.G.C. Philip, L.W. Philip, B. Michael,
E.B. Donald, T.B. Brian, B. Georgiy, C. Libing, C. Chih-Yuan, H.C. Thomas,
F. Zahra, F. Wenlang, R.K. Uday, A.K. James, L. Xiao-Gao, B.L. Derek,
C.M. Andrea, M. Martin, A.O. Astrid, D.R. Philip, W.S. Robert, E.S. Tatiana,
V. Alexandros, T. Weifeng, W. Lei, Y. Lin, J.G. Stephen, N.L. James, J.S. Laurel,
H.B. William, J. Med. Chem. 50 (2007) 5202–5216.
[18] B. Atsuo, K. Noriaki, M. Haruhiko, O. Yoshikazu, T. Shigehisa, S. Takashi, J. Med.
Chem. 39 (1996) 5176–5182.
[19] J.C. Medina, M.G. Johanson, A. Li, J. Lu, A.X. Huang, L. Zhu, A.P. Marcus, PCT Int
Appl. WO. 02 83, 143; Chem. Abstr. 137 (2002) 337909e.
[20] B.Y. Daniel, W.G. Jason, L.N. Stephanie, V.P. Arely, T.C. Matthew, A.B. David, J.
Dermatol. Sci. 42 (2006) 13–21.
[21] R.J. VanRyzin, J.H. Trpold, Drug Chem. Toxicol. 3 (4) (1980) 361–379.
[22] D. Wheathly, Rheumatol. Rehabil. 21 (2) (1982) 98–100.
[23] S. Fankhauser, W. Laube, H.R. Marti, H.R. Schultheiss, J. Vo¨gtlin, B. von Graf-
fenried, Arzneimittelforschung 31 (5a) (1981) 934–935.
[24] O. Cheristine, H. Matthias, Planta Med. 70 (2004) 642–645.
[25] A. Kumar, R.S. Verma, B.P. Jaju, J.N. Sinha, J. Indian Chem. Soc. 67 (1990) 920–921.
[26] A.A. Bekhit, H.M.A. Ashour, Y.S. Abdel Ghany, A.A. Bekhit, A. Baraka, Eur. J. Med.
Chem. 43 (2008) 456–463.
[27] D. Villemin, L. Liang, Tetrahydron Lett. 37 (48) (1996) 8733–8734.
[28] K. Kawasaki, I. Yamada, T. Oe, M. Tsuruda, M. Terasawa, T. Imayoshi,
Y. Yasunaga, K. Goto, Yakugaku Zasshi 109 (11) (1989) 827–834.
[29] A.K. Gadad, B.S. Kittur, S.G. Kapsi, C.S. Mahajanshetti, S.B. Rajur, Arznei-
mittelforschung 46 (1996) 1082–1085.
5.2.5. Ulcerogenic effects
Groups of 5 male Wistar rats with a weight between 150 and
175 g are used. They are starved 48 h prior to drug administration.
The test compounds are administered orally in 10 mL/kg as aqueous
suspension. Doses are chosen which are highly active in the activity
(9 mg/kg) and used. The animals are sacrificed after 7 h. Stomachs
are removed and placed on saline soaked filter paper until inspec-
tion. A longitudinal incision along the greater curvature is made
with fine scissor. The stomach is inverted over the index finger and
the presence or the absence of gastric irritation is determined. The
presence of a single or multiple lesions (erosion, ulcer or perfora-
tion) is considered to be positive [49]. The number of ulcers and the
occurrence of hyperemia is noted (determine ulcer index).
5.3. Molecular modeling
All computational data were performed on Intel (R) Core (TM) 2
DUO CPU, 2.6 GHZ, 2.27 GHZ. With memory (RAM) ¼ 4.00 GB and
has Ubunto 8.04 release software. Docking was done by Plants
software [50]and autodock vina [51,52].
[30] K. Tsuji, K. Nakamura, T. Ogino, N. Konishi, T. Tojo, T. Ochi, N. Seki, M. Matsuo,
Chem. Pharm. Bull. 46 (1998) 279–286.
[31] S.A. Beers, E.A. Malloy, W. Wu, M. Wachter, J. Ansell, M. Singer, M. Steber,
A. Barbone, T. Kirchner, D. Ritchie, D. Argentieri, Bioorg. Med. Chem. Lett. 5
(1997) 779–786.
Acknowledgement
[32] D. Venu, S.
A Khnum, F. Aiysha, B.K. Manuprasad, S. Sheena, M. Riyaz,
S.V. Bannikuppe, Bioorg. Med. Chem. Lett. 18 (2008) 4409–4412.
[33] M. Salimi, M.H. Ghahremani, N. Naderi, M. Amini, M. Salimi, M. Amanlou,
K. Abdi, R. Salehi, A. Shafiee, Acta Pharmacol. Sin 28 (8) (2007) 1254–1260.
[34] B. Goel, T. Ram, R. Tyagi, E. Banasal, A. Kumar, D. Mukerjee, J.N. Shina, Eur. J.
Med. Chem. 34 (1999) 265–269.
[35] K. Srivastava, R. Yadav, S.D. Srivastava, Indian J. Chem. 42B (2004) 399–405.
[36] M. Suma, S.K. Srivastava, S.D. Srivastava, Indian J. Chem. 36B (1997) 826–830.
[37] O.M. Fathalla, E.M.M. Kassem, N.M. Ibrahem, M.M. Kamel, Acta Pol. Pharm.
Drug Res. 65 (1) (2008) 11–20.
[38] G.M. Badger, R.W. Hinde, R.W. Matthews, T.M. Spots-wood, Aust. J.Chem. 16
(1963) 732–733.
[39] P.R. Carlier, Y.F. Han, E.S. Chow, T.X. Lieu, H.S. Wong, Y.P. Pang, Bioorg. Med.
Chem. 7 (1999) 351–357.
The authors thank the Micro-analytical Unit, Cairo University,
Egypt for micro-analytical data, IR, 1H NMR and Mass spectra.
References
[1] C. Bombardier, L. laine, A. Reicin, D. Shapiro, R. Burgos-Vargas, B. Davis, R. Day,
M. Bosi Ferraz, C.J. Hawkey, M.C. Hochberg, T.K. Kvien, T.J.N. Schnitzer, Eng. J.
Med. 343 (2000) 1520–1528.
[2] F.E. Silverstien, G. Faich, J.L. Goldstien, L.S. Simon, T. Pincus, A. Whelton,
R. Makuch, G. Eisen, N.M. Agrawal, W.F. Stenson, A.M. Burr, W.W. Zhao,
J.D. Kent, J.B. Lefkowith, K.M. Verburg, G.S. Gies, J. Am. Med. Assoc. 284 (2000)
1247–1255.
[3] S. Mark, E. Jan, A. Sabrina, L. Martina, A. Silke, R. Yeon, A. Jodi, S. Gillian,
R. William, M. Detlef, E. Stephen, K. Bert, S. Thomas, M. Manfred, Antivir. Res.
79 (2008) 49–61.
[40] S. Kim, C. Ahn, S. Keum, K. Koh, Dyes Pigm. 65 (2005) 179–182.
[41] P.P. Yadav, P. Gupta, A.K. Chaturvedi, P.K. Skukla, R. Maurya, Bioorg. Med. Chem.
13 (2005) 1497–1505.
[42] N.A. Hamdy, Egypt. J. Chem. 48 (6) (2005) 749–758.
[43] E.M. Kassem, A.H. Mandour, Egypt. J. Chem. 42 (4) (1999) 387–401.
[44] M.Y. Ebied, M.M. Kamel, Z.M. Nofal, F. Ragab, W.A. Zaghary, M. Elkady, Egypt. J.
Chem. 49 (2) (2006) 277–285.
[4] L. Kubicova, P. Kudelova, H. Dostal, K. Waisser, Folia Pharm. Univ. Carol 25
(2000) 81–87.
[5] J. Hanusek, Chem. Listy 95 (2001) 811–813.
[45] F.D. Popp, J. Med. Pharm. Chem. 5 (1962) 627–629.
[6] H.J. Chan, J.S. Hong, L.F. Kuyper, D.P. Baccanari, S.S. Joyner, R.L. Tansik,
C.M. Boytos, S.K. Rudolph, J. Med. Chem. 38 (1995) 3608–3616.
[7] R. Castaldo, D. Gump, Mc. Cormack, J. Antimicrob. Agents Chemother. 15
(1979) 81–86.
[46] G.A. Winter, E.A. Rislfy, G.W. Nuss, Proc. Soc. Exp. Biol. Med.111 (1962) 544–547.
[47] P. Armitage, Statistical Methods in Medical Research, first ed. Blackwell
Scientific Publ., Oxford, London, 1971, 147.
[8] K. Harushia, Y. Kiesuke, H. Seiko, H. Shingo, K. Ryota, H. Norimitsu, M. Makoto,
O. Yoshiteru, J. Med. Chem. 49 (15) (2006) 4698–4706.
[9] T. Yasutaka, S. Takao, W. Nobuhisa, A. Hideyuki, S. Shigeru, S. Isao, J. Med.
Chem. 37 (1994) 2106–2111.
[48] G. Woolfe, A.D. MacDonald, J. Pharmacol. Exp. Ther. 80 (1944) 300–307.
[49] A.E. Amr, M.M. Abdulla, Arch. Pharm. Chem. Life Sci. 339 (2006) 88–95.
[50] O. Korb, T.E. Stutzle, An ant colony optimization approach to flexible protein-
ligand docking. Swarm Intelligence 1 (2007) 115–134.
[51] O. Trott, A.J. Olson, Autodock vina: improving the speed of docking with a new
scoring function, efficient optimization and multithreading. Journal of
Computational Chemistry, in press.
[52] F. Michel. Sanner, Python: a programming language for software integration
and development. J. Mol. Graph. Model 17 (1999) 57–61.
[53] M.G. Obukowics, D.J. Walseh, W.J. Salsgiver, C.L. Martin-Berger, K.S. Chinn,
K.L. Duffin, A. Ras, P. Needleman, J. Pharmacol. Exp. Ther. 287 (1998) 157–166.
[54] L. Meng, R. Mohan, B.H.B. Kwok, M. Elofssof, N. Sin, C.M. Crews, Proc. Natl.
Acad. Sci. USA 96 (1999) 10403–10408.
[10] D. Marianne, P. Fredric, C. Olivier, T. Jean-Claude, C. Jean-Pierre, B. Yves, Chem.
Pharm. Bull. 49 (9) (2001) 1061–1065.
[11] P. Mani Chandrica, T. Yakaiah, A. Raghu Ram Rao, B. Narsaiah, N. ChakraReddy,
V. Sridhar, J. Venkateshwara Rao, Eur. J. Med. Chem. 43 (2008) 846–852.
[12] M.H. Yen, J.R. Sheu, I.H. Peng, Y.M. Lee, J.W. Chern, J. Pharm. Pharmacol. 48 (1)
(1996) 90–95.
[13] S. Hyao, M.J. Mvera, W. Strycker, T. Leipzi, R. Klup, H. Hartzler, J. Med. Chem. 8
(1965) 807–811.
[14] E. Cohen, E. Klarberg, R. JamesVaughan Jr., J. Am. Chem. Soc. 82 (11) (1960)
2731–2735.
[55] M. Eaton, J. Rehabil. Res. Dev. 40 (2003) 41–54.