New quinazolinone-pyrimidine hybrids: Synthesis, anti-inflammatory, and ulcerogenicity studies

Article abstract of DOI:10.1016/j.ejmech.2012.03.050

Two groups of hybrid compounds: the quinazolinone-dihydropyrimidines and quinazolinone-pyrimidines, were synthesized. The starting derivative 3 was reacted with chloroacetyl chloride to give intermediate 5 which was condensed with the 2-mercaptopyrimidines 4a-c affording compounds 6a-c. These latter compounds underwent hydrolysis and N-alkylation reactions to give the dihydropyrimidine derivatives 7a-c and 8a-f, respectively. The chloro derivatives 9a-c subsequently reacted with various anilines furnishing compounds 10a-i. The anti-inflammatory activity of the synthesized compounds were evaluated using the carrageenan-induced rat paw oedema model and ulcer indices for the most active compounds were calculated. Five compounds were found more active and less ulcerogenic than diclofenac particularly compound 10g (IC ₅₀ = 116.73 μmol/kg; ulcer index = 11.38). Compound 10g was also 2-fold more selective inhibitor of COX-2 than COX-1.

Full text of DOI:10.1016/j.ejmech.2012.03.050

European Journal of Medicinal Chemistry 53 (2012) 141e149
Contents lists available at SciVerse ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
New quinazolinoneepyrimidine hybrids: Synthesis, anti-inflammatory,
and ulcerogenicity studies
,
b
b
c
Safinaz E. Abbas a, Fadi M. Awadallah a , Nashwa A. Ibrahin , Eman G. Said , Gihan M. Kamel
*
a Pharmaceutical Chemistry Department, Faculty of Pharmacy, Cairo University, 11562 Kasr El-Eini, Cairo, Egypt
b Pharmaceutical Chemistry Department, Faculty of Pharmacy, Beni-Suef University, 62111 Beni-Suef, Egypt
c Pharmacology Department, Faculty of Veterinary Medicine, Cairo University, Egypt
a r t i c l e i n f o
a b s t r a c t
Article history:
Two groups of hybrid compounds: the quinazolinoneedihydropyrimidines and quinazolinoneepyrimidines,
were synthesized. The starting derivative 3 was reacted with chloroacetyl chloride to give intermediate 5
which was condensed with the 2-mercaptopyrimidines 4aec affording compounds 6aec. These latter
compoundsunderwent hydrolysisand N-alkylationreactions to give the dihydropyrimidine derivatives 7aec
and 8aef, respectively. The chloro derivatives 9aec subsequently reacted with various anilines furnishing
compounds 10aei. The anti-inflammatory activity of the synthesized compounds were evaluated using the
carrageenan-induced rat paw oedema model and ulcer indices for the most active compounds were calcu-
lated. Five compounds were found more active and less ulcerogenic than diclofenac particularly compound
Received 28 January 2012
Received in revised form
22 March 2012
Accepted 26 March 2012
Available online 9 April 2012
Keywords:
Quinazolinone
Pyrimidine
Dihydropyrimidine
Anti-inflammatory activity
COX-1/COX-2
10g (IC50 ¼ 116.73
m
mol/kg; ulcer index ¼ 11.38). Compound 10g was also 2-fold more selective inhibitor of
COX-2 than COX-1.
Ó 2012 Elsevier Masson SAS. All rights reserved.
Ulcerogenicity
1. Introduction
term prophylactic use in certain chronic diseases [4,10,11]. This has
led intense efforts in search for potent and selective COX-2 inhibi-
Inflammation is a multifactorial process. It reflects the response
of the organism to various stimuli and is related to many disorders
such as arthritis, asthma and psoriasis which require prolonged or
repeated treatment [1]. It is well known that non steroidal anti-
inflammatory drugs (NSAIDs) are associated with several side
effects such as gastrointestinal mucosal damage, bleeding, intoler-
ance and renal toxicity [2,3]. The pharmacological effects of NSAIDs
are due to inhibition of a membrane enzyme called cyclo-oxygenase
(COX) which is involved in the prostaglandin biosynthesis [4]. There
are two isoforms, COX-1 and COX-2. The constitutive COX-1 plays
a physiological role in the kidneys and the stomach, whereas, the
mainly inducible COX-2 is involved in the production of prosta-
glandins mediating pain and supporting the inflammatory process
[5e7]. Gastric injury associated with classical NSAIDs is attributed
to the nonselective inhibition of the two isoenzymes [8,9]. In order
to prevent or decrease this side effect, a current strategy consists of
designing selective COX-2 inhibitors with an improved gastric
safety profile which may allow the use of these new agents for long-
tors which could provide anti-inflammatory drugs with fewer risks.
The quinazoline nucleus is considered an important chemical
synthon of various physiological significance and pharmaceutical
utility. Quinazolines have drawn a great attention due to their wide
range of therapeutic activities including antiviral [12], antibacterial
[13,14], antifungal [15,16], antimalarial [17], anticancer [18e20],
antihypertensive [21], diuretic [22,23], inhibition of derived growth
factor receptor phosphorylation [24], anticonvulsant [25], antago-
nism of ghrelin receptor [26], anti-inflammatory, analgesic and
COX-2 inhibitory activities [27e29]. The well known anti-
inflammatory drug, proquazone I [30], and the recently devel-
oped derivatives II [31] and III [32], are good examples of quina-
zolinone derivatives with potent anti-inflammatory activity (Fig. 1).
In addition, literature survey revealed that many effective anti-
inflammatory agents were derived from the pyrimidine, dihy-
dropyrimidine and tetrahydropyrimidine nuclei as shown in
compounds IV [33], V [34] and VI [35], respectively (Fig. 2).
In the view of the facts mentioned above, and in an attempt to
design and develop new potential anti-inflammatory agents,
a hybrid pharmacophoric approach was adopted in which the
quinazolinone and substituted pyrimidine/dihydropyrimidine
moieties were hybridized in one structure hoping to synergize the
* Corresponding author. Tel.: þ20 2 26070436; þ20 12 23483941.
E-mail address: fadi_mae@hotmail.com (F.M. Awadallah).
0223-5234/$ e see front matter Ó 2012 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejmech.2012.03.050

142
S.E. Abbas et al. / European Journal of Medicinal Chemistry 53 (2012) 141e149
S
O
S
SO2CH3
N
O
N
O
N
H
O
N
N
N
N
O
N
H
OCH3
Proquazon I
II
III
Fig. 1. Examples of some quinazolinone-derived anti-inflammatory agents.
anti-inflammatory potential of both groups. The validity of this
design was assessed through preliminary in vivo anti-inflammatory
screening and ulcerogenicity study of the target compounds.
in two magnetically unequivalent environments since they
appeared as one upfield proton as doublet at 5.01 and 4.78 ppm and
one downfield proton as doublet at 5.31 and 4.92 ppm, respectively.
The carboxylic acid derivatives 7aec were obtained upon hydrolysis
of their carbonitrile precursors 6aec using 80% sulphuric acid. IR
spectra of compounds 7aec revealed the disappearance of CN band
and alternatively broad carboxylic OH band appeared in the range of
3400e3141 cmꢀ1. N-alkylation of 6aec with the appropriate alkyl
halide in absolute ethanol in the presence of potassium carbonate
afforded the derivatives 8aef. The target compounds were essen-
tially characterized by their 1H NMR spectra that showed the signals
assigned to the NeCH3 protons of compounds 8a, 8c and 8e as
singlets at 3.24, 3.60 and 3.34, respectively. Likewise, 1H NMR
spectra of compounds 8b, 8d and 8f revealed the benzylic CH2
protons as singlets at 5.68, 5.08 and 5.28, respectively together with
the extra aromatic protons of the benzyl group.
The second group of compounds, the pyrimidinyl-quinazoline
derivatives 9aec and 10aei, was prepared through chlorination
of 6aec with phosphorous oxychloride to give the chloro inter-
mediates 9aec which were reacted with the appropriate anilines in
absolute ethanol affording the target anilino derivatives 10aei
(Scheme 3). Compounds 10aei were characterized by their 1H
NMR spectra that declared an increase in the integration of
aromatic protons attributed to the anilino aromatic protons
together with the exchangeable anilino NHs.
2. Results and discussion
2.1. Chemistry
Two groups of target compoundswere prepared;thefirst included
the dihydropyrimidinyl-quinazolinone derivatives 6aec, 7aec and
8aef and the second consisted of the pyrimidinyl-quinazolinone
derivatives 9aec and 10aei. The starting compound 3-amino-2-
(2,4-dichlorophenoxy)methyl-3,4-dihydroquinazolin-4-one
3 was
prepared from methyl anthranilate 1 which was reacted with 2,4-
dichlorophenoxyacetyl chloride to give the intermediate 2 followed
by reflux with hydrazine hydrate in n-butanol (Scheme 1) [36].
The first group of compounds was prepared as shown in Scheme
3. Reaction of 3 with chloroacetyl chloride gave the chloroacetyl
derivative 5 [37]. The dihydropyrimidinyl compounds 6aec were
obtained through S-alkylation of the appropriate 2-mercapto-6-
oxo-4-(un)substitutedphenyl-1,6-dihydropyrimidine-5-carbonit
riles 4aec with the chloro intermediate 5 in dry acetone in the
presence of potassium carbonate. Compounds 4aec were prepared
as depicted in Scheme 2 following the reported procedure [38].
Primary evidence for the formation of the target compounds 6aec
was drawn from IR spectra that revealed a CN stretching band at
about 2200 cmꢀ1. 1H NMR spectrum 6c revealed the CH2S protons
as singlet at 4.37 ppm, the CH2O protons as singlet at 5.09 ppm, in
addition to the other aromatic and exchangeable protons appearing
as multiplet at 7.29e7.93 ppm. The proton at C-5 of quinazoline ring
appeared separately as doublet at 8.15 ppm. It was also noted that
the two protons of CH2O group in compounds 6a and 6b were lying
2.2. Anti-inflammatory screening
Evaluation of the anti-inflammatory activity of the synthesized
compounds was performed using the carrageenan-induced rat paw
oedema model using diclofenac sodium as the reference drug
[39e42]. Mean changes in paw oedema thickness of animals
OCH3
OCH3
O
HN
N
O
N
NC
CH3
HOOC
NH
C2H5O
HN
N
N
N
N
H
S
N
Cl
H2N
COOC2H5
V
IV
VI
Fig. 2. Examples of some pyrimidine-, dihydropyrimidine- and tetrahydropyrimidine-based anti-inflammatory agents.

S.E. Abbas et al. / European Journal of Medicinal Chemistry 53 (2012) 141e149
143
O
N
O
O
NH2
O
CH3
O
CH3
N
O
b
a
Cl
NH
Cl
NH2
O
O
3
1
2
Cl
Cl
Scheme 1. Synthesis of compound 3. Reagents and conditions. a: 2,4-dichlorophenoxyacetyl chloride, dry ether, TEA, rt. 24 h; b: hydrazine hydrate (85%), n-butanol, reflux 10 h.
pretreated with the tested compounds after 0.5, 1, 2 and 3 h from
inductionofinflammationwasmeasured,togetherwiththeinhibition
percent of oedema by the tested compounds (Table 1).
rings, compound 10a. The IC50 (mmol/kg) of these five compounds
were calculated and presented in Table 2. In terms of IC50, all
compounds except 6a were more active than diclofenac sodium.
Also, it could be observed that the 4-anilino derivatives (10f,10g and
10i) were more active than both 8e and 6a. The most active
The preliminary anti-inflammatory activity of the first group of
compounds comprising the dihydropyrimidinyl-quinazolinone
derivatives showed that unsubstituted phenyl group at position 4
of the dihydropyrimidine ring, compound 6a, was befitting to high
activity. This held true in the corresponding carboxyl derivatives
7aec in which compound 7a exhibited higher activity than 7b and
7c. Regarding the introduction of a substituent on N1 of the dihy-
dropyrimidine ring, activity of 6a decreased when either a methyl
group, compound 8a, or a benzyl group, compound 8b, were
introduced. The opposite held true for compound 6b whose activity
increased by the presence of an N-substitution as in compounds 8c
and 8d. However, with compound 6c there was no consistent
relation since the activity increased in the N-methyl derivative 8e
and decreased in the N-benzyl derivative 8f.
compound was 10f having an IC50 of 114.03 mmol/kg.
2.3. Ulcerogenicity study
Compounds 6a, 8b, 8d, 8e, 10a, 10fei with best overall profile in
animal efficacy model were evaluated for gastric ulcerogenic
potential in rats (Table 3) [43,44]. Ulcerogenic effect was compared
to a classical NSAID, diclofenac sodium and to a non-ulcerogenic
drug, celecoxib. Results revealed that all of the tested compounds
were less ulcerogenic than diclofenac sodium and that compounds
8b, 8d, 8e, 10g and 10h were less ulcerogenic than celecoxib.
In the pyrimidinyl-quinazolinone derivatives, the presence of the
4-chlorogroupon the pyrimidineringofcompounds9b and 9c, unlike
9a, was associated with higher activity compared to their precursors
6b and 6c. As per the 4-(un)substitutedanilino-pyrimidine derivatives
10aei, the presence of an unsubstituted anilino group, compound 10a,
positively affected the activity compared to its precursor 9a; while the
reverse was true with the substituted anilino moieties. The activity of
9b was greatly enhanced with the p-hydroxyanilino group as shown
by derivative 10f, which exhibited an activity similar to diclofenac.
Regarding compound 9c, introduction of an (un)substituted anilino
group at position 4 of the pyrimidine ring contributed to the increase
in activity in all derivatives 10gei. It is noteworthy that compound 10g
was equipotent to diclofenac.
2.4. COX-2/COX-1 selectivity assay
Compounds 6a, 8e, 10f, 10g, 10i were evaluated for their
selectivity to inhibition of COX-2 and COX-1 isoenzymes using COX
(ovine) inhibitor screening assay kit according to the manufac-
turer’s instructions. The ratio of IC50 of COX-2 to IC50 of COX-1
(COX-2/COX-1) would suggest the selectivity of the compound
and hence its gastric liability. The COX-2/COX-1 ratio of the above
compounds showed that compounds 8e, 10f and 10g are selective
COX-2 inhibitor with a ratio of 0.62, 0.40 and 0.44, respectively
(Table 4).
3. Conclusion
In conclusion, five, out of all synthesized compounds, showed
potency above 90% of that of diclofenac: 6a, 8e, 10f, 10g and 10i. In
particular, the 4-anilinopyrimidine derivatives 10f and 10g were
equipotent to diclofenac. The highest overall activity was observed
in the 4-anilinopyrimidinyl-quinazolinone series 10aei, especially
in all compounds possessing 4-hydroxyphenyl group (electron
donating, hydrophilic), compounds 10gei, or having a 4-chloro
anilino group (electron withdrawing, lipophilic) compromised by
the 6-(4-hydroxyphenyl) ring (electron donating, hydrophilic),
compound 10f, or unsubstituted on both the 4-anilino and 6-phenyl
Two sets of quinazolinone hybrids were prepared: the
dihydropyrimidinyl-quinazolinone
and
the
pyrimidinyl-
quinazolinone hybrids which were tested for their anti-
inflammatory activity and ulcerogenic potential. Preliminary activity
revealed that the pyrimidinyl-quinazolinones possessing an anilino
group in position 4 of the pyrimidine ring were the most active
especially compounds 10fei. The dihydropyrimidinyl derivatives were
generally less active except for compounds 6a and 8e. These
compounds possessed IC50 values and ulcer indices lower than
R1
O
CN
S
HN
HS
ethanol
K2CO3
H3C
O
+
+
H2N
NH2
CN
N
O
CHO
R
4a R= H
4b R=Cl
4c R=OH
Scheme 2. Synthesis of compounds 4aec.

144
S.E. Abbas et al. / European Journal of Medicinal Chemistry 53 (2012) 141e149
O
N
CN
O
O
N
O
N
HN
H
N
H
N
NH2
O
N
Cl
a
N
b
N
S
O
O
N
Cl
Cl
Cl
R
O
O
Cl
Cl
Cl
3
5
e
6a R= H
6b R= Cl
6c R= OH
c
Cl
O
N
CN
d
O
N
N
COOH
H
N
O
N
HN
H
N
N
S
N
N
S
O
Cl
R
O
Cl
R
O
O
O
N
Cl
R'
CN
Cl
O
N
N
9a R= H
9b R=Cl
H
N
N
S
7a R= H
7b R= Cl
9c R= OH
O
f
7c R= OH
Cl
R
O
Cl
R'
8a R= H; R'= CH3
8b R= H; R'= CH2-C6H5
8c R= Cl; R'= CH3
NH
N
CN
8d R= Cl; R'= CH2-C6H5
8e R= OH; R'= CH3
O
N
H
N
N
S
8f R= OH; R=' CH2-C6H5
O
N
Cl
R
O
Cl
10a R= H; R'= H
10b R= H; R'= Cl
10c R= H; R'= OH
10d R= Cl; R'= H
10e R= Cl; R'= Cl
10f R= Cl; R'= OH
10g R= OH; R'= H
10h R= OH; R'= Cl
10i R= OH; R'= OH
Scheme 3. Synthesis of compounds 6aec, 7aec, 8aef, 9aec and 10aei. Reagents and conditions. a: chloroacetyl chloride, dry benzene, TEA, reflux, 4 h; b: compounds 4aec, dry
acetone, anhydrous K2CO3, reflux, 6 h; c: sulphuric (80%), stir, 24 h; d: alkyl halide, absolute ethanol, anhydrous K2CO3, reflux, 8 h; e: phosphorous oxychloride, stir, rt, 1/2 h, reflux,
6 h; f: appropriate aniline, absolute ethanol, stir, rt, 24 h then reflux, 5 h.
diclofenac and compounds 8e and 10g were even less ulcerogenic than
4. Experimental
celecoxib at the same dose level. The most active and safest compound
is 10g (IC50 ¼ 116.73
m
mol/kg; ulcer index ¼ 11.38). Also, COX-2/COX-1
4.1. Chemistry
selectivity assay suggested that compounds 8e, 10f and 10g were 2-
fold selective inhibitors to COX-2 than COX-1. According to these
results, it could be claimed that our aim was achieved through the
development of compounds more active and of less ulcerogenic side
effects than standard drugs.
Melting points were uncorrected and were carried out by open
capillary tube method using IA 9100 MK-Digital Melting Point
Apparatus. Elemental microanalyses were carried out on Heraew
and Vario El III (elementar), CHNS analyzer (Germany) at the

S.E. Abbas et al. / European Journal of Medicinal Chemistry 53 (2012) 141e149
145
Table 1
Oedema thickness and inhibition percent in oedema thickness of control, diclofenac sodium and tested compounds.
Compound
Nꢁ
Oedema thickness (mm) ꢂ SEM (oedema inhibition %)
0.5 h
1 h
2 h
3 h
Control
Diclofenac sodium
6a
6b
6c
7a
7b
7c
8a
8b
8c
8d
8e
8f
9a
9b
0.520 ꢂ 0.023
0.550 ꢂ 0.022
0.570 ꢂ 0.024
0.570 ꢂ 0.024
0.120 ꢂ 0.008c (76.92)
0.100 ꢂ 0.006c (80.77)
0.200 ꢂ 0.009c*** (61.54)
0.200 ꢂ 0.013c*** (61.54)
0.200 ꢂ 0.014c*** (61.54)
0.250 ꢂ 0.012c*** (51.92)
0.350 ꢂ 0.018c*** (32.69)
0.250 ꢂ 0.020c*** (51.92)
0.125 ꢂ 0.008c (75.96)
0.100 ꢂ 0.007c (80.77)
0.200 ꢂ 0.01c*** (61.54)
0.120 ꢂ 0.01c (76.92)
0.090 ꢂ 0.004c (83.64)
0.100 ꢂ 0.007c (81.82)
0.200 ꢂ 0.009c*** (63.64)
0.200 ꢂ 0.013a*** (63.64)
0.170 ꢂ 0.012c*** (69.09)
0.230 ꢂ 0.014a*** (58.18)
0.350 ꢂ 0.018a*** (36.36)
0.250 ꢂ 0.020c*** (54.55)
0.125 ꢂ 0.008c** (77.27)
0.130 ꢂ 0.009a** (76.36)
0.150 ꢂ 0.006c*** (72.73)
0.110 ꢂ 0.010 (80.00)
0.070 ꢂ 0.005c (87.72)
0.080 ꢂ 0.004c*** (85.96)
0.200 ꢂ 0.008c*** (64.91)
0.180 ꢂ 0.012a (68.42)
0.180 ꢂ 0.013c*** (68.42)
0.220 ꢂ 0.01a (61.40)
0.050 ꢂ 0.003c (91.23)
0.080 ꢂ 0.004c*** (85.96)
0.200 ꢂ 0.008c** (64.91)*
0.180 ꢂ 0.012** (68.42)*
0.200 ꢂ 0.015c*** (64.91)
0.220 ꢂ 0.01*** (61.40)
0.350 ꢂ 0.017*** (38.59)
0.230 ꢂ 0.018c*** (59.65)
0.120 ꢂ 0.009*** (78.95)
0.150 ꢂ 0.01*** (73.68)
0.130 ꢂ 0.008a (77.19)
0.080 ꢂ 0.006** (85.96)
0.220 ꢂ 0.013*** (61.40)
0.180 ꢂ 0.012c*** (68.41)
0.180 ꢂ 0.007*** (68.42)
0.150 ꢂ 0.07c*** (73.68)
0.125 ꢂ 0.007*** (78.07)
0.230 ꢂ 0.016*** (59.65)
0.230 ꢂ 0.015*** (59.65)
0.250 ꢂ 0.02c*** (56.14)
0.200 ꢂ 0.016c*** (64.91)
0.050 ꢂ 0.003c (91.23)
0.050 ꢂ 0.003c (91.23)
0.120 ꢂ 0.01c*** (78.95)
0.075 ꢂ 0.004a*** (86.84)
0.350 ꢂ 0.017a (38.59)
0.230 ꢂ 0.020c*** (59.65
0.120 ꢂ 0.009a*** (78.95
0.150 ꢂ 0.009a*** (73.68)
0.130 ꢂ 0.008a*** (77.19)
0.080 ꢂ 0.006c (85.96)
0.250 ꢂ 0.012a*** (56.14)
0.190 ꢂ 0.012c*** (66.67)
0.180 ꢂ 0.008a (68.42)
0.150 ꢂ 0.07c*** (73.68
0.125 ꢂ 0.007a*** (78.07)
0.240 ꢂ 0.015a (57.89)
0.230 ꢂ 0.016a (59.65)
0.280 ꢂ 0.022c*** (50.88)
0.220 ꢂ 0.021c*** (61.40)
0.050 ꢂ 0.003a (91.23)
0.050 ꢂ 0.003a** (91.23)
0.130 ꢂ 0.012c** (77.19)
0.075 ꢂ 0.004c (86.84)
0.250 ꢂ 0.012c*** (51.92)
0.240 ꢂ 0.016c*** (53.85)
0.200 ꢂ 0.013c*** (61.54)
0.170 ꢂ 0.011c** (67.31)
0.150 ꢂ 0.009c* (71.15)
0.250 ꢂ 0.013c*** (51.92)
0.200 ꢂ 0.012c*** (61.54)
0.330 ꢂ 0.023c*** (36.54)
0.280 ꢂ 0.023c*** (46.15)
0.150 ꢂ 0.008c (71.15)
0.050 ꢂ 0.003c*** (90.38)
0.180 ꢂ 0.02c* (65.38)
0.100 ꢂ 0.007c (80.77)
0.250 ꢂ 0.012a*** (54.55)
0.220 ꢂ 0.014c*** (60.00)
0.200 ꢂ 0.013a*** (63.64)
0.150 ꢂ 0.08c*** (72.73)
0.125 ꢂ 0.007a** (77.27)
0.250 ꢂ 0.013a*** (54.55)
0.220 ꢂ 0.013a*** (72.73)
0.310 ꢂ 0.024c*** (43.64)
0.250 ꢂ 0.022c*** (54.55)
0.050 ꢂ 0.003a*** (90.91)
0.050 ꢂ 0.003c*** (90.91)
0.150 ꢂ 0.016c** (72.73)
0.075 ꢂ 0.005c* (86.36)
9c
10a
10b
10c
10d
10e
10f
10g
10h
10i
Values with superscript letter a significantly different with control value at P ꢃ 0.05, c are significantly different with control value at P ꢃ 0.001, values with stars are
significantly different with diclofenac-sodium values * at P ꢃ 0.05, ** at P ꢃ 0.01 and *** at P ꢃ 0.001. Results are mean ꢂ S.E. of five animals.
Microanalytical Center, Faculty of Science, Cairo University. Infrared
spectra were made on Bruker Vector 22 (Japan), infrared spectro-
photometers and were expressed in wave number (cmꢀ1) using
potassium bromide disc. 1H NMR spectra were recorded on a Varian
Mercury VX-300 NMR spectrometer at 300 MHz and 13C NMR were
recorded at 75 MHz in the specified solvent. Chemical shifts were
reported on the d scale and were related to that of the solvent and J
values are given in Hz. Mass spectra were recorded on Shimadzu
Qp-2010 plus, at 70 eV (EI). IUPAC chemical nomenclature were
assigned using CS Chemdraw ultra version 5.0. Thin layer chro-
matography was performed using MachereyeNagel Alugram Sil G/
UV254 silica gel plates and petroleum ether: ethyl acetate (9:1) as
the eluting system.
4.1.1. General procedure for the preparation of compounds (6aec)
A mixture of the chloro derivative 5 (4.1 g, 10 mmol), the
appropriate 2-mercapto-dihydropyrimidine derivative 4aec
(10 mmol) and anhydrous potassium carbonate (1.37 g, 10 mmol)
was refluxed in dry acetone (20 ml) for 6 h. The reaction mixture
was filtered while hot and the residue was washed with hot
acetone. The organic solution was concentrated and the separated
crude product was filtered off and recrystallized from
chloroformeethanol.
Table 2
IC50 values of the anti-inflammatory activity for compounds 6a, 8e, 10f, 10g, 10i and
diclofenac sodium.
Compound Nꢁ
Dose(mg/kg)
Oedema inhibition
% after 3 h
IC50 mg/kg
(IC50 mmol/kg)
4.1.1.1. 2-(5-Cyano-6-oxo-4-phenyl-1,6-dihydropyrimidin-2-ylsulfanyl)
-N-[2-(2,4-dichlorophenoxymethyl)-4-oxo-4H-quinazolin-3-yl]acet-
Diclofenac-sodium
20
50
100
42.25
87.26
91.23
45.01 (141.03)
86.74 (143.26)
82.89 (130.44)
83.36 (114.03)
81.31 (116.73)
88.27 (123.88)
amide (6a). Yellow powder; m.p. 170e172 ꢁC, yield, 60%. IR nmax
/
6a
50
100
200
68.42
85.96
91.23
Table 3
Ulcer index of tested compounds of the most active compounds, diclofenac sodium
and celecoxib.
8e
50
100
200
70.42
85.96
97.75
Compound
Nꢁ
No. of animals
with ulcer
% Incidence
/10
Average no
of ulcer
Average
severity
Ulcer
index
Control
Diclofenac-
sodium
Celecoxib
6a
8b
8d
8e
10a
10f
10g
10h
10i
0/5
5/5
0
10
0
5.60
0
1.42
0
17.02
10f
10g
10i
50
100
200
61.40
91.23
96.47
4/5
4/5
4/5
4/5
4/5
4/5
5/5
4/5
4/5
5/5
8
8
8
8
8
8
10
8
8
10
3.20
5.20
2.20
2.80
2.60
4.20
3.60
3.00
3.40
3.40
1.31
1.20
0.70
0.50
0.62
1.06
0.83
0.38
0.68
0.82
12.51
14.40
10.90
11.30
11.22
13.26
14.43
11.38
12.08
14.22
50
100
200
77.19
91.23
96.49
50
100
200
56.14
86.84
91.23

146
S.E. Abbas et al. / European Journal of Medicinal Chemistry 53 (2012) 141e149
Table 4
4.1.2.2. 4-(4-Chlorophenyl)-2-{[2-(2,4-dichlorophenoxymethyl)-4-oxo-
4H-quinazolin-3-ylcarbamoyl]-methylsulfanyl}-6-oxo-1,6-dihydropyri-
midine-5-carboxylic acid (7b). Buff powder; m.p. 265e267 ꢁC, yield,
60%. IR nmax/cmꢀ1: 3400 (br., OH, NHs); 3095 (CH aromatic); 2922 (CH
COX-2/COX-1 ratio of compounds 6a, 8e, 10f, 10g, 10i and
celecoxib.
Compound
COX-2/COX-1
aliphatic); 1607 (COs); 772 (CeCl). 1H NMR (DMSO-d6):
d 4.28 (s, 2H,
6a
8e
10f
10g
10i
Celecoxib
20.60
0.62
0.40
0.44
7.30
0.02
CH2S); 4.72 (s, 2H, CH2O); 6.74e7.80 (m, 10H, aromatic H); 8.13 (d, 1H,
C5-H of quinazoline, J ¼ 7.9 Hz); 11.40 (s, 2H, 2 NHs exchangeable with
D2O); 11.45 (s, 1H, COOH, exchangeable). Anal. Calcd. for
C28H18Cl3N5O6S (658.90): C, 51.04; H, 2.75; N, 10.63. Found: C, 51.26;
H, 2.90; N, 10.01.
4.1.2.3. 2-{[2-(2,4-Dichlorophenoxymethyl)-4-oxo-4H-quinazolin-3-
ylcarbamoyl]-methylsulfanyl}-4-(4-hydroxyphenyl)-6-oxo-1,6-dihy-
dropyrimidine-5-carboxylic acid (7c). Buff powder; m.p. 245e247 ꢁC,
yield, 62%. IR nmax/cmꢀ1: 3141 (br., OHs, NHs); 3085 (CH aromatic);
cmꢀ1: 3308 (NHs); 3198 (CH aromatic); 2923 (CH aliphatic); 2216
(C^N); 1681 (COs); 1608, 1477 (C]N, C]C); 769 (CeCl). 1H NMR
(DMSO-d6):
d 3.69 (s, 2H, CH2S); 5.01(d, 1H, upfield proton of CH2O,
J ¼ 16.2 Hz); 5.31(d, 1H, downfield proton of CH2O, J ¼ 16.2 Hz);
7.05e7.76 (m, 13H, 11 aromatic H and 2 NHs exchangeable); 8.06 (d,
1H, C5-H of quinazoline, J ¼ 7.8 Hz). MS, m/z (%): 606 (M þ 1, 3.03);
161 (100). Anal. Calcd. for C28H18Cl2N6O4S (605.45): C, 55.55; H, 3.00;
N, 13.88. Found: C, 55.50; H, 3.39; N, 14.01.
2929 (CHaliphatic); 1613 (COs); 769 (CeCl).1H NMR (DMSO-d6):
d 4.14
(s, 2H, CH2S); 5.14 (s, 2H, CH2O); 6.82e7.82 (m, 14H, 10 aromatic H, 2
NHs and 2 OHs exchangeable); 8.26 (d, 1H, C5-H of quinazoline,
J ¼ 7.8 Hz). Anal. Calcd. for C28H19Cl2N5O7S (640.45): C, 52.51; H, 2.99;
N, 10.94. Found: C, 52.59; H, 3.29; N, 11.28.
4.1.1.2. 2-[4-(4-Chlorophenyl)-5-cyano-6-oxo-1,6-dihydropyrimidin-
2-ylsulfanyl]-N-[2-(2,4-dichlorophenoxymethyl)-4-oxo-4H-quinazo-
lin-3-yl]acetamide (6b). Yellow powder; m.p. 190e192 ꢁC, yield,
70%. IR nmax/cmꢀ1: 3414 (NHs); 3076 (CH aromatic); 2927 (CH
aliphatic); 2215 (C^N); 1689 (COs); 1611, 1551 (C]N, C]C); 772
4.1.3. General procedure for the preparation of compounds (8aef)
Methyl iodide/benzyl chloride (10 mmol) was added to
a mixture of the appropriate compound 6aec (10 mmol) and
anhydrous potassium carbonate (2.1 g, 15 mmol) in absolute
ethanol (20 ml). The reaction mixture was refluxed for 8 h. The
solvent was evaporated under reduced pressure and the residue
was treated with water, filtered and the crude product was crys-
tallized from chloroformeethanol.
(CeCl). 1H NMR (DMSO-d6):
d 4.25 (s, 2H, CH2S); 4.78(d, 1H upfield
proton of CH2O, J ¼ 15.9 Hz); 4.92(d, 1H, downfield proton of CH2O,
J ¼ 15.6 Hz); 6.84e7.95 (m, 10H, aromatic H); 8.13 (d, 1H, C5-H of
quinazoline, J ¼ 7.95 Hz); 11.40 (s, 2H, 2NHs exchangeable). 13C
NMR (DMSO-d6):
d
32.5 (CH2S); 65.8 (CH2O); 115.0e130.2
4.1.3.1. 2-(5-Cyano-1-methyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-
2-ylsulfanyl)-N-[2-(2,4-dichlorophenoxymethyl)-4-oxo-4H-quinazo-
lin-3-yl]acetamide (8a). Brown powder; m.p. 260e262 ꢁC, yield,
70%. IR nmax/cmꢀ1: 3368 (NH); 3194 (CH aromatic); 2918 (CH
aliphatic); 2207 (C^N); 1633 (COs); 768 (CeCl).1H NMR (DMSO-d6):
(aromatic C); 135.2 (C-1 chlorophenyl); 145.8 (C-1 dichlor-
ophenyl); 152.2 (C-2 pyrimidine); 158.4 (C]O quinazoline); 164.0
(C-2 quinazoline); 165.6 (C]O pyrimidine); 168.1 (C]O carbox-
amide). Anal. Calcd. for C28H17Cl3N6O4S (639.90): C, 52.56; H, 2.68;
N, 13.13. Found: C, 52.60; H, 2.71; N, 13.16.
d
3.24 (s, 3H, CH3); 4.63 (s, 2H, CH2S); 5.37 (s, 2H, CH2O); 7.10e7.94
(m, 11H, aromatic H); 8.30 (d, 1H, C5-H of quinazoline, J ¼ 8.1 Hz);
11.2 (s, 1H, NH exchangeable). Anal. Calcd. for C29H20Cl2N6O4S
(619.48): C, 56.23; H, 3.25; N,13.57. Found: C, 56.32; H, 3.70; N,14.01.
4.1.1.3. 2-[5-Cyano-4-(4-hydroxyphenyl)-6-oxo-1,6-dihydropyrimid-
in-2-ylsulfanyl]-N-[2-(2,4-dichlorophenoxymethyl)-4-oxo-4H-quina-
zolin-3-yl]acetamide (6c). Off-White crystals; m.p. 200e202 ꢁC,
yield, 55%. IR nmax/cmꢀ1: 3404 (br., OH, NHs); 3108 (CH aromatic);
2918 (CH aliphatic); 2197 (C^N); 1607 (COs); 1555 (C]N, C]C);
4.1.3.2. 2-(1-Benzyl-5-cyano-6-oxo-4-phenyl-1,6-dihydropyrimidin-
2-ylsulfanyl)-N-[2-(2,4-dichlorophenoxymethyl)-4-oxo-4H-quinazo-
lin-3-yl]acetamide (8b). Light-brown powder; m.p. 190e192 ꢁC,
yield, 69%. IR nmax/cmꢀ1: 3419 (NH); 3040 (CH aromatic); 2916 (CH
aliphatic); 2213 (C^N); 1669, 1605 (COs); 762 (CeCl). 1H NMR
771 (CeCl). 1H NMR (DMSO-d6):
CH2O); 7.29e7.93 (m, 13H, 10 aromatic H,
d
4.37 (s, 2H, CH2S); 5.09 (s, 2H,
NHs and OH
2
exchangeable); 8.15 (d, 1H, C5-H of quinazoline, J ¼ 7.8 Hz). Anal.
Calcd. for C28H18Cl2N6O5S (621.45): C, 54.12; H, 2.92; N, 13.52.
Found: C, 53.82; H, 3.05; N, 13.20.
(DMSO-d6): d 4.76 (s, 2H, CH2S); 5.50 (s, 2H, CH2O); 5.68 (s, 2H, CH2-
C6H5); 7.00e7.83 (m, 17H, 16 aromatic H and NH exchangeable); 8.15
(d,1H, C5-H of quinazoline, J ¼ 8.1 Hz). Anal. Calcd. for C35H24Cl2N6O4S
(695.58): C, 60.44; H, 3.48; N,12.08. Found: C, 60.98; H, 3.91; N,12.48.
4.1.2. General procedure for the preparation of compounds (7aec)
A mixture of the cyano derivatives 6aec (6.02 mmol) in 80%
sulphuric (5 ml) was stirred at room temperature for 24 h. The
reaction mixture was diluted with cold water, neutralized with
diluted ammonia solution and filtered. The separated solid was
dried and crystallized from ethanol.
4.1.3.3. 2-[4-(4-Chlorophenyl)-5-cyano-1-methyl-6-oxo-1,6-dihydro
pyrimidin-2-ylsulfanyl]-N-[2-(2,4-dichlorophenoxymethyl)-4-oxo-4H-
quinazolin-3-yl]acetamide (8c). Brown powder; m.p. 200e202 ꢁC,
yield, 73%. IR nmax/cmꢀ1: 3435 (NH); 3091 (CH aromatic); 2924 (CH
aliphatic); 2219 (C^N); 1687 (COs); 777 (CeCl). 1H NMR (DMSO-d6):
4.1.2.1. 2-{[2-(2,4-Dichlorophenoxymethyl)-4-oxo-4H-quinazolin-3-
ylcarbamoyl]-methylsulfanyl}-6-oxo-4-phenyl-1,6-dihydropyrimidine-
5-carboxylic acid (7a). Orange powder; m.p. 250e252 ꢁC, yield, 55%.
IR nmax/cmꢀ1: 3374 (br., OH, NHs); 3041 (CH aromatic); 2923 (CH
d 3.60 (s, 3H, CH3); 5.05 (s, 2H, CH2S); 5.42 (s, 2H, CH2O); 7.19e7.93
(m, 10H, aromatic H); 8.13 (d, 1H, C5-H of quinazoline, J ¼ 6.9 Hz);
11.44 (s, 1H, NH exchangeable). Anal. Calcd. for C29H19Cl3N6O4S
(653.92): C, 53.26; H, 2.93; N, 12.85. Found: C, 53.23; H, 3.22; N,
12.55.
aliphatic); 1662 (COs); 770 (CeCl).1H NMR (DMSO-d6):
d 5.10 (s, 2H,
CH2S); 5.80 (s, 2H, CH2O); 7.30e7.76 (m, 11H, aromatic H); 8.15 (d, 1H,
C5-H of quinazoline, J ¼ 7.8 Hz); 10.05 (s, 3H, 2 NHs and COOH
exchangeable). MS, m/z (%): 625 (M þ 1, 3.53); 627 ((M þ 3), 1.71); 77
(100). Anal. Calcd. for C28H19Cl2N5O6S (624.45): C, 53.86; H, 3.07; N,
11.22. Found: C, 54.04; H, 3.37; N, 10.88.
4.1.3.4. 2-[1-Benzyl-4-(4-chlorophenyl)-5-cyano-6-oxo-1,6-dihydro-
pyrimidin-2-ylsulfanyl]-N-[2-(2,4-dichlorophenoxymethyl)-4-oxo-4H-
quinazolin-3-yl]acetamide (8d). Orange powder; m.p. 220e222 ꢁC,
yield, 70%. IR nmax/cmꢀ1: 3436 (NH); 3105 (CH aromatic); 2924 (CH

S.E. Abbas et al. / European Journal of Medicinal Chemistry 53 (2012) 141e149
147
aliphatic); 2209 (C^N); 1686 (COs); 1548, 1467 (C]N, C]C); 774
(CeCl). 1H NMR (DMSO-d6):
3.91 (d, 1H, upfield proton of CH2S,
downfield proton of CH2O, J ¼ 11.4 Hz); 6.90e8.08 (m, 10H,
aromatic H); 8.23 (d, 1H, C5-H of quinazoline, J ¼ 8.1 Hz); 9.09 (s,
1H, NH exchangeable). Anal. Calcd. for C28H16Cl4N6O3S (658.34): C,
51.08; H, 2.45; N, 12.77. Found: C, 51.50; H, 2.11; N, 12.31.
d
J ¼ 15 Hz); 4.04 (d, 1H, downfield proton of CH2S, J ¼ 15.9 Hz); 4.94 (d,
1H, upfield proton of CH2O, J ¼ 15.6 Hz); 5.08 (s, 2H, CH2-C6H5); 5.14
(d, 1H, downfield proton of CH2O, J ¼ 15.6 Hz); 6.94e7.86 (m, 15H,
aromatic H); 8.12 (d, 1H, C5-H of quinazoline, J ¼ 7.8 Hz); 11.38 (s, 1H,
4.1.4.3. 2-(4-Chloro-5-cyano-6-(4-hydroxyphenyl)pyrimidin-2-ylsul
fanyl)-N-[2-(2,4-dichlorophenoxymethyl)-4-oxo-4H-quinazolin-3-yl]
NH exchangeable). 13C NMR (DMSO-d6):
d 32.5 (CH2S); 41.2 (CH2
benzyl); 65.9 (CH2O); 115.2e129.8 (aromatic C); 135.8 (C-1 chlor-
ophenyl); 145.9 (C-1 dichlorophenyl); 152.3 (C-2 pyrimidine); 158.4
(C]O quinazoline); 164.1 (C-2 quinazoline); 165.6 (C]O pyrimi-
dine); 169.0 (C]O carboxamide). Anal. Calcd. for C35H23Cl3N6O4S
(730.02): C, 57.58; H, 3.18; N, 11.51. Found: C, 57.25; H, 3.33; N, 11.28.
acetamide (9c). Buff powder; m.p. 225e227 ꢁC, yield, 55%. IR nmax
/
cmꢀ1: 3369 (br., OH, NH); 3084 (CH aromatic); 2922 (CH aliphatic);
2208 (C^N); 1633 (COs); 769 (CeCl). 1H NMR (CDCl3):
4.72 (d, 1H,
d
upfield proton of CH2S, J ¼ 16.2 Hz); 4.92 (d, 1H, downfield proton
of CH2S, J ¼ 16.5 Hz); 5.34 (s, 2H, CH2O); 7.16e7.87 (m, 10H,
aromatic H); 8.23 (d, 1H, C5-H of quinazoline, J ¼ 7.8 Hz); 9.09 (s,
1H, NH exchangeable with D2O); 11.64 (s, 1H, OH exchangeable).
Anal. Calcd. for C28H17Cl3N6O4S (639.90): C, 52.56; H, 2.68; N, 13.13.
Found: C, 52.67; H, 2.88; N, 12.85.
4.1.3.5. 2-[5-Cyano-4-(4-hydroxyphenyl)-1-methyl-6-oxo-1,6-dihyd
ropyrimidin-2-ylsulfanyl]-N-[2-(2,4-dichlorophenoxymethyl)-4-oxo-
4H-quinazolin-3-yl]acetamide
(8e). Brown
powder;
m.p.
235e237 ꢁC, yield, 65%. IR nmax/cmꢀ1: 3282 (br., NH, OH); 3085 (CH
aromatic); 2927 (CH aliphatic); 2230 (C^N); 1702 (COs); 1589,1512
4.1.5. General procedure for the preparation of compounds (10aei)
To a stirred solution of the appropriate aniline (10 mmol) and
triethylamine (0.5 ml) in absolute ethanol (10 ml), a solution of
compound 9aec (10 mmol) in absolute ethanol (10 ml) was added
portion wise. The reaction mixture was stirred for 24 h at room
temperature, and then refluxed for 5 h. The solvent was removed by
distillation under vacuum, the residue was triturated with cold
water and the solid was filtered off and crystallized from methanol.
(C]N); 1473 (C]C); 770 (C-Cl). 1H NMR (DMSO-d6):
d 3.34 (s, 3H,
CH3); 4.29 (s, 2H, CH2S); 5.09 (s, 2H, CH2O); 6.92e8.23 (m, 12H, 10
aromatic H, NH and OH exchangeable); 8.10 (d, 1H, C5-H of qui-
nazoline, J ¼ 8.0 Hz). Anal. Calcd. for C29H20Cl2N6O5S (635.48): C,
54.81; H, 3.17; N, 13.22. Found: C, 55.07; H, 3.03; N, 12.85.
4.1.3.6. 2-[1-Benzyl-5-cyano-4-(4-hydroxyphenyl)-6-oxo-1,6-dihydr
opyrimidin-2-ylsulfanyl]-N-[2-(2,4-dichlorophenoxymethyl)-4-oxo-
4H-quinazolin-3-yl]acetamide
(8f). Brown
powder;
m.p.
4.1.5.1. 2-(5-Cyano-6-phenyl-4-phenylaminopyrimidin-2-ylsulfanyl)
-N-[2-(2,4-dichlorophenoxymethyl)-4-oxo-4H-quinazolin-3-yl]acet-
240e242 ꢁC, yield, 60%. IR nmax/cmꢀ1: 3377 (OH); 3280 (NH); 3051
(CH aromatic); 2926 (CH aliphatic); 2214 (C^N); 1683, 1603 (COs);
amide (10a). Brown powder; m.p. 225e227 ꢁC, yield, 60%. IR nmax
/
1506 (C]N); 1476 (C]C); 770 (CeCl). 1H NMR (DMSO-d6):
d
4.24 (s,
cmꢀ1: 3393 (NHs); 3058 (CH aromatic); 2925 (CH aliphatic); 2210
2H, CH2S); 5.07 (s, 2H, CH2O, J ¼ 3.9 Hz); 5.28 (s, 2H, CH2-C6H5);
(C^N); 1697 (COs); 763 (CeCl). 1H NMR (CDCl3):
d
3.03 (s, 2H, CH2S),
6.57e7.82 (m, 17H, 15 aromatic H, NH and OH exchangeable); 8.20
4.68 (s, 2H, CH2O), 6.91e7.98 (m, 18H, 16 aromatic H and 2 NHs
exchangeable); 8.79 (d, 1H, C5-H of quinazoline, J ¼ 7.9 Hz). 13C NMR
(d, 1H, C5-H of quinazoline, J ¼ 8.2 Hz). 13C NMR (DMSO-d6):
d 32.6
(CH2S); 40.0 (CH2 benzyl); 65.8 (CH2O); 115.8e131.3 (aromatic C);
135.1 (C-1 chlorophenyl); 144.8 (C-1 dichlorophenyl); 153.8 (C-2
pyrimidine); 155.7 (C-4 hydroxyphenyl); 158.3 (C]O quinazoline);
163.8 (C-2 quinazoline); 165.5 (C]O pyrimidine); 168.1 (C]O
carboxamide). Anal. Calcd. for C35H24Cl2N6O5S (635.48): C, 59.08;
H, 3.40; N, 11.81. Found: C, 59.25; H, 3.70; N, 12.03.
(DMSO-d6): d 32.1 (CH2S); 65.9 (CH2O); 115.3e130.7 (aromatic C);
142.2 (C-1 anilino); 149.5 (C-1 dichlorophenyl); 160.3 (C]O quina-
zoline); 163.7 (C-2 quinazoline); 170.3 (C]O carboxamide); 172.5 (C-
4 pyrimidine); 174.1 (C-2 pyrimidine). MS, m/z (%): 681 (Mþ, 0.57); 63
(100). Anal. Calcd. for C34H23Cl2N7O3S (680.56): C, 60.00; H, 3.41; N,
14.41. Found: C, 60.29; H, 3.89; N, 14.11.
4.1.4. General procedure for the preparation of compounds (9aec)
Compound 6aec (11.1 mmol) was added portion wise with stir-
ring to ice-cooled phosphorous oxychloride (10 ml). The reaction
mixture was stirred at room temperature for 30 min. The mixture
was then heated to reflux for 6 h. The cooled reaction mixture was
poured on crushed ice and the separated solid was filtered off,
washed with water, dried and crystallized from aqueous ethanol.
4.1.5.2. 2-[4-(4-Chlorophenylamino)-5-cyano-6-phenylpyrimidin-2-
ylsulfanyl]-N-[2-(2,4-dichlorophenoxymethyl)-4-oxo-4H-quinazolin-
3-yl]acetamide (10b). Buff powder; m.p. 230e232 ꢁC, yield, 65%. IR
nmax/cmꢀ1: 3398 (NHs); 3063 (CH aromatic); 2924 (CH aliphatic);
2213 (C^N); 1697 (COs); 764 (CeCl). 1H NMR (CDCl3):
d 3.72 (s, 2H,
CH2S), 4.98 (s, 2H, CH2O), 6.86e7.80 (m, 15H, aromatic H); 8.01 (d,
1H, C5-H of quinazoline, J ¼ 7.8 Hz); 11.90 (s, 2H, NHs exchange-
able). Anal. Calcd. for C34H22Cl3N7O3S (715.01): C, 57.11; H, 3.10; N,
13.71. Found: C, 56.89; H, 3.45; N, 14.04.
4.1.4.1. 2-(4-Chloro-5-cyano-6-phenylpyrimidin-2-ylsulfanyl)-N-[2-
(2,4-dichlorophenoxymethyl)-4-oxo-4H-quinazolin-3-yl]acetamide
(9a). Yellow crystals; m.p. 230e232 ꢁC, yield, 57%. IR nmax/cmꢀ1
:
4.1.5.3. 2-[5-Cyano-4-(4-hydroxyphenylamino)-6-phenylpyrimidin-
2-ylsulfanyl]-N-[2-(2,4-dichlorophenoxymethyl)-4-oxo-4H-quinazo-
lin-3-yl]acetamide (10c). Buff crystals; m.p. 235e237 ꢁC, yield, 55%.
IR nmax/cmꢀ1: 3385 (br., OH, NHs); 3065 (CH aromatic); 2929 (CH
aliphatic); 2211 (C^N); 1688 (COs); 764 (CeCl).1H NMR (DMSO-d6):
3240 (NH); 3067 (CH aromatic); 2916 (CH aliphatic); 2230 (C^N);
1697 (COs); 760 (CeCl). 1H NMR (CDCl3):
d
4.12 (s, 2H, CH2S); 4.73
(s, 2H, CH2O); 6.92e8.13 (m, 11H, aromatic H); 8.81 (d, 1H, C5-H of
quinazoline, J ¼ 8.4 Hz); 11.62 (s, 1H, NH exchangeable). MS, m/z
(%): 624((M)þ, 2.27); 625 ((M þ 1), 2.07); 162(100). Anal. Calcd. for
C28H17Cl3N6O3S (623.90): C, 53.90; H, 2.75; N, 13.47. Found: C,
53.63; H, 2.32; N, 13.95.
d
4.20 (s, 2H, CH2S); 5.58 (s, 2H, CH2O); 6.42e7.85 (m, 15H, aromatic
H); 8.16 (d, 1H, C5-H of quinazoline, J ¼ 8.1 Hz); 9.62 (s, 3H, 2 NHs
and OH exchangeable). Anal. Calcd. for C34H23Cl2N7O4S (696.56): C,
58.63; H, 3.33; N, 14.08. Found: C, 58.24; H, 3.65; N, 14.28.
4.1.4.2. 2-(4-Chloro-6-(4-chlorophenyl)-5-cyanopyrimidin-2-ylsul
fanyl)-N-[2-(2,4-dichlorophenoxymethyl)-4-oxo-4H-quinazolin-3-yl]
acetamide (9b). Yellow crystals; m.p. 240e242 ꢁC, yield, 60%. IR
nmax/cmꢀ1: 3309 (NH); 3077 (CH aromatic); 2921 (CH aliphatic);
4.1.5.4. 2-[6-(4-Chlorophenyl)-5-cyano-4-phenylaminopyrimidin-2-
ylsulfanyl]-N-[2-(2,4-dichlorophenoxymethyl)-4-oxo-4H-quinazolin-
3-yl]acetamide (10d). Brown powder; m.p. 235e237 ꢁC, yield, 65%.
IR nmax/cmꢀ1: 3394, 3298 (NHs); 3054 (CH aromatic); 2924 (CH
aliphatic); 2212 (C^N); 1695 (COs); 756 (CeCl). 1H NMR (DMSO-
2200 (C^N); 1678 (COs); 771 (CeCl). 1H NMR (CDCl3):
d
4.11 (s, 2H,
CH2S); 4.85 (d, 1H, upfield proton of CH2O, J ¼ 12 Hz); 4.95 (d, 1H,

148
S.E. Abbas et al. / European Journal of Medicinal Chemistry 53 (2012) 141e149
d6):
d
4.19 (s, 2H, CH2S); 4.68 (d, 1H upfield proton of CH2O,
(DMSO-d6): d 4.94 (s, 2H, CH2S); 5.22 (s, 2H, CH2O); 7.15e7.96 (m,
J ¼ 16.5 Hz); 4.81 (d, 1H, downfield proton of CH2O, J ¼ 16.4 Hz);
6.81e8.13 (m, 15H, aromatic H); 8.57 (d, 1H, C5-H of quinazoline,
J ¼ 7.95 Hz); 11.20 (s, 1H, NH, exchangeable with D2O); 11.30 (s, 1H,
NH, exchangeable). Anal. Calcd. for C34H22Cl3N7O3S (715.01): C,
57.11; H, 3.10; N, 13.71. Found: C, 57.09; H, 3.22; N, 13.44.
14H, aromatic H); 8.16 (d, 1H, C5-H of quinazoline, J ¼ 8.3 Hz); 11.24
(s, 2H, 2 NHs exchangeable); 13.93 (s, 2H, 2 OHs exchangeable). Anal.
Calcd. for C34H23Cl2N7O5S (712.56): C, 57.31; H, 3.25; N,13.76. Found:
C, 57.35; H, 3.72; N, 13.80.
4.2. Pharmacological screening
4.1.5.5. 2-[6-(4-Chlorophenyl)-4-(4-chlorophenylamino)-5-cyanop-
yrimidin-2-ylsulfanyl]-N-[2-(2,4-dichlorophenoxymethyl)-4-oxo-4H-
quinazolin-3-yl]acetamide (10e). Orange crystals; m.p. 240e242 ꢁC,
yield, 57%. IR nmax/cmꢀ1: 3410, 3307 (NHs); 3075 (CH aromatic); 2924
(CH aliphatic); 2213 (C^N); 1689, 1606 (COs); 772 (CeCl). 1H NMR
4.2.1. Anti-inflammatory activity
Adult albino rats of both sexes weighing between 120 and 150 g
were used. Rats were uniformly hydrated by giving 3 ml water/rat
through gastric inoculation to reduce variability to oedema
response. Animals were divided into 26 groups each of five animals.
The control group was given saline solution containing few drops of
Tween 80. Diclofenac sodium (50 mg/kg) was taken as standard
drug for comparison and compounds under examination (100 mg/
kg) were suspended in distilled water by the aid of few drops of
Tween 80 and were given orally 1 h before induction of inflam-
mation. Induction of inflammation was performed by S.C. injection
(CDCl3):
d
3.95 (d, 1H upfield proton of CH2S, J ¼ 11.1 Hz); 4.03 (d, 1H,
downfield proton of CH2S, J ¼ 12.4 Hz); 4.72 (d, 1H upfield proton of
CH2O, J ¼ 12.5 Hz); 4.80 (d, 1H, downfield proton of CH2O,
J ¼ 13.4 Hz); 6.76e8.02 (m, 14H, aromatic H); 8.26 (d, 1H, C5-H of
quinazoline, J ¼ 8.2 Hz); 9.24 (s, 2H, NHs exchangeable). Anal. Calcd.
for C34H21Cl4N7O3S (749.45): C, 54.49; H, 2.82; N, 13.08. Found: C,
54.40; H, 2.52; N, 12.89.
of 50 ml of 1% carrageenan-sodium gel (SigmaeAldrich, USA), into
4.1.5.6. 2-[6-(4-Chlorophenyl)-5-cyano-4-(4-hydroxyphenylamino)
pyrimidin-2-ylsulfanyl]-N-[2-(2,4-dichlorophenoxymethyl)-4-oxo-
the sub-plantar region of the right hind paw. The dorso-ventral
diameter (thickness) of the right and left hind paw of each rat
was measured using a pair of dial thickness gauge callipers accurate
to 0.001 cm 0.5, 1, 2 and 3 h after induction of inflammation. The
left hind paw diameter served as a control for the degree of
inflammation in the right hind paw. The percentage of anti-
inflammatory activity (% inhibition of inflammation) was calcu-
lated according to the following equation:
4H-quinazolin-3-yl]acetamide
(10f). Brown
powder;
m.p.
242e244 ꢁC, yield, 55%. IR nmax/cmꢀ1: 3391 (OH); 3297 (NHs); 3055
(CH aromatic); 2924 (CH aliphatic); 2211 (C^N); 1691 (COs); 763
(CeCl). 1H NMR (DMSO-d6):
d 3.22 (s, 2H, CH2S); 4.91 (s, 2H, CH2O);
6.79e7.93 (m, 15H, aromatic H); 10.5 (s, 1H, NH exchangeable with
D2O); 11.32 (s, 1H, NH exchangeable); 11.50 (s, 1H, OH exchange-
able). Anal. Calcd. for C34H22Cl3N7O4S (731.01): C, 55.86; H, 3.03; N,
13.41. Found: C, 56.02; H, 3.27; N, 13.46.
% inhibition ¼ ðWc ꢀ Wt=WcÞ ꢄ 100
4.1.5.7. 2-[5-Cyano-6-(4-hydroxyphenyl)-4-phenylaminopyrimidin-
2-ylsulfanyl]-N-[2-(2,4-dichlorophenoxymethyl)-4-oxo-4H-quinazo-
lin-3-yl]acetamide (10g). Light-brown powder; m.p. 245e247 ꢁC,
yield, 55%. IR nmax/cmꢀ1: 3370, 3200 (OH, NHs); 3076 (CH
aromatic); 2956 (CH aliphatic); 2221 (C^N); 1693 (COs); 768 (C-
Wt: is the mean increase in paw thickness in rats treated with
the tested compounds.
Wc: is the mean increase in paw thickness in control group.
Cl). 1H NMR (DMSO-d6):
d 4.33 (s, 2H, CH2S); 5.11 (s, 2H, CH2O);
6.96e7.79 (m, 15H, aromatic H); 8.21 (d, 1H, C5-H of quinazoline,
J ¼ 8.1 Hz); 8.59 (s, 1H, NH exchangeable); 8.60 (s, 1H, NH
exchangeable with D2O); 9.90 (s,1H, OH exchangeable). Anal. Calcd.
for C34H23Cl2N7O4S (696.56): C, 58.63; H, 3.33; N, 14.08. Found: C,
59.01; H, 3.24; N, 13.99.
Data were analyzed by SPSS statistical package version 10.
Results are presented in Table 1.
4.2.2. Acute ulcerogenicity study
Adult albino rats of both sexes weighing between 120 and 150 g
were used. Animals were divided into groups each of five animals.
Rats were fasted 20 h before drug administration. The tested
compounds, diclofenac sodium and celecoxib were given orally in
a dose of 100, 50 and 50 mg/kg, respectively, suspended in 1%
Tween while one group received vehicle (1% Tween). Rats were
fasted for 2 h, allowed to feed for 2 h then fasted for another 20 h.
Rats were given another two doses in the second and third days. In
the fourth day, rats were sacrificed, the stomach removed, opened
along with the greater curvature and rinsed with 0.9% saline. The
number of mucosal damage (red spots) was counted and their
severity (ulcerogenic severity) was graded from 0 to 4 according to
the following score assignment: The following figures were
calculated:
4.1.5.8. 2-[4-(4-chlorophenylamino)-5-cyano-6-(4-hydroxyphenyl)
pyrimidin-2-ylsulfanyl]-N-[2-(2,4-dichlorophenoxymethyl)-4-oxo-
4H-quinazolin-3-yl]acetamide
(10h). Brown
powder;
m.p.
250e252 ꢁC, yield, 53%. IR nmax/cmꢀ1: 3373 (OH, NHs); 3075 (CH
aromatic); 2923 (CH aliphatic); 2215 (C^N); 1694, 1609 (COs); 773
(CeCl). 1H NMR (CDCl3):
d 4.72 (d, 1H, upfield proton of CH2S,
J ¼ 12.6 Hz); 5.08 (d, 1H, downfield proton of CH2S, J ¼ 12.3 Hz);
5.33 (s, 2H, CH2O); 6.60e7.80 (m, 17H, 14 aromatic H, 2 NHs and OH
exchangeable); 8.25(d, 1H, C5-H of quinazoline, J ¼ 8.2 Hz). 13C
NMR (DMSO-d6):
d 32.3 (CH2S); 65.7 (CH2O); 114.3e132.8
(aromatic C); 140.1 (C-1 chloroanilino); 146.8 (C-1 dichlor-
ophenyl); 156.7 (C-4 hydroxyphenyl); 158.3 (C]O quinazoline);
163.8 (C-2 quinazoline); 170.1 (C]O carboxamide); 172.6 (C-4
pyrimidine); 174.6 (C-2 pyrimidine). Anal. Calcd. for
C34H22Cl3N7O4S (731.01): C, 55.86; H, 3.03; N, 13.41. Found: C,
55.55; H, 2.92; N, 13.05.
- % Incidence/10 ¼ [number of rats showing ulcer of any grade
divided by total number of rats in the group ꢄ 100]/10.
- Average number of ulcers: number of ulcers in the group/total
number of rats in the group.
P
4.1.5.9. 2-[5-Cyano-6-(4-hydroxyphenyl)-4-(4-hydroxyphenylamino)
pyrimidin-2-ylsulfany]-N-[2-(2,4-dichlorophenoxymethyl)-4-oxo-4H-
quinazolin-3-yl]acetamide (10i). Buff powder; m.p. 245e247 ꢁC,
yield, 50%. IR nmax/cmꢀ1: 3396 (OHs, NHs); 3095 (CH aromatic); 2941
(CH aliphatic); 2212 (C^N); 1639 (COs); 789 (CeCl). 1H NMR
- Average severity:
severity]/number of ulcers in the group.
[each ulcer multiplied by its score of
Ulcer index ¼ the sum of the 3 figures
Results are tabulated in Table 3.

S.E. Abbas et al. / European Journal of Medicinal Chemistry 53 (2012) 141e149
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