Synthesis and molecular modeling studies of anti-inflammatory active 1H-pyrrolizine-5-carboxamides

Article abstract of DOI:10.1002/ardp.201000166

A variety of N-aryl-7-cyano-2,3-dihydro-1H-pyrrolizine-5-carboxamides 5, 6, 8, and 9 were synthesized via reaction of the 2-amino derivatives 4 with acid chlorides and aromatic aldehydes. Meanwhile, 4a,b were obtained through the reaction of 2-pyrrolidiny

Full text of DOI:10.1002/ardp.201000166

56
Arch. Pharm. Chem. Life Sci. 2011, 1, 56–65
Full Paper
Synthesis and Molecular Modeling Studies of Anti-
inflammatory Active 1H-Pyrrolizine-5-carboxamides
Flora F. Barsoum
Pharmaceutical Chemistry Department, Faculty of Pharmacy, Cairo University, Cairo, Egypt
A variety of N-aryl-7-cyano-2,3-dihydro-1H-pyrrolizine-5-carboxamides 5, 6, 8, and 9 were synthesized
via reaction of the 2-amino derivatives 4 with acid chlorides and aromatic aldehydes. Meanwhile, 4a,b
were obtained through the reaction of 2-pyrrolidinylidenepropanedinitrile 1 with chloroacetanilides
2a,b. In addition, the tricyclic pyrimido[5,4-a]pyrrolizines were formed through conducting the
reaction of 4a,b with 90% formic acid. Anti-inflammatory activity screening of some synthesized
compounds utilizing in vivo acute carrageenan-induced paw edema standard method in rats exhibited
that the prepared heterocycles possess considerable pharmacological properties especially, 4a, 4b,
10a, and 10b which reveal remarkable activities relative to diclofenac sodium (reference standard).
Ulcerogenic liability of the highly promising synthesized anti-inflammatory active agents were
evaluated and 4a and 4b showed ulcerogenic liability lower than that of the standard used drug.
Molecular modeling studies were initiated herein in order to validate the attained pharmacological
data and provide understandable evidence for the observed anti-inflammatory behavior.
Keywords: Anti-inflammatory / Chloroacetanilides / Molecular modeling/ 1H-Pyrrolizine-5-carboxamides / Ulcerogenic
liability
Received: June 5, 2010; Revised: July 1, 2010; Accepted: July 12, 2010
DOI 10.1002/ardp.201000166
Introduction
anti-inflammatory derivatives (ImSAIDs) that are class of pep-
tides altering the activation and migration of inflammatory
cells which are responsible for amplifying the inflammatory
response [8, 9]; in addition to medical drugs, some herbs have
anti-inflammatory qualities and can reduce the inflam-
mation [10–12].
Inflammation is a complex biological response of vascular
tissues against harmful stimuli, such as pathogens, damaged
cells or irritants, mediated by different physiological and
immunological mediators and characterized by the accumu-
lation of fluids and leukocytes leading to edema and pain [1].
Acute inflammation occurs as the initial response to tissue
injury, being mediated by the release of autacoids as hista-
mine, bradykinin, prostaglandins and leukotrienes. On the
other hand, the chronic inflammatory process involves the
release of diverse mediators, as interleukins, interferon and
tumor necrosis factor a (TNF-a) [2]. Drugs currently used for
reducing inflammation include; steroids, specifically gluco-
corticoids by binding to cortisol receptors [3–5]; non-steroidal
anti-inflammatory drugs (NSAIDs), alleviate pain by counter-
acting the cyclooxygenase enzyme [6, 7]; immune selective
Non-steroidal anti-inflammatory drugs are commonly
used for the treatment of pain and inflammation, however,
long-term therapy may cause gastrointestinal complications
ranging from stomach irritation to life-threatening gastro-
intestinal ulceration and bleeding [13, 14]. Several pharma-
cological drugs have been synthesized but none of them
is free of side effects, so it is important to find new anti-
inflammatory drug with a potential for clinical use and not
associated with adverse effects. Several bi- and tricyclic com-
pounds have been synthesized using the starting material
2-pyrrolidinylidenepropanedinitrile and they possess anti-
inflammatory and analgesic activities [15–20], therefore with
the aim of obtaining new anti-inflammatory drugs with
greater activity and fewer side effects, in the work described
herein, we intended to investigate the synthesis of new
Correspondence: Flora F. Barsoum, Pharmaceutical Chemistry
Department, Faculty of Pharmacy, Cairo University, 11562, Cairo, Egypt
E-mail: ffbarsoum2@yahoo.com
ß 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Arch. Pharm. Chem. Life Sci. 2011, 1, 56–65
Anti-inflammatory 1H-Pyrrolizine-5-carboxamides
57
bi- and tricyclic compounds via a facile synthetic approach
using the same starting material and evaluating their anti-
inflammatory activity. Ulcerogenic properties of the most
anti-inflammatory active synthesized compounds will be also
considered and a structure-activity relationship analysis was
also done in order to understand the structural requirements
for optimum activity.
substitution reaction yielded the corresponding pyrrolizines
1
9a,b,d in good yields. H-NMR spectra of 9a,b,d exhibit the
acetamido methylene protons upfield shifted (d ¼ 3.29, 3.31,
3.55) compared with the parent chloroacetamido methylene
function of 8a,c which appeared at d ¼ 4.26, 4.31. Similarly,
the reaction of 4a,b with 3-chloropropionyl chloride 7b gave
the chloropropionylamino-1H-pyrrolizine derivatives 8b,d
which underwent nucleophilic substitution reaction with
secondary amines, giving the corresponding pyrrolizines
9c,e in good yields.
Results and discussion
Chemistry
Moreover, refluxing 4a,b with 90% formic acid afforded the
tricyclic pyrimido[5,4-a]pyrrolizine-9-carboxamides 10a,b
through intramolecular cyclization onto the cyano group.
The structure of 10 was established through spectroscopic (IR,
1H-NMR) as well as elemental analyses data. The IR spectra
reveal two bands attributed to the two carbonyl groups at
n ¼ 1699–1658 cmꢀ1 region and the absence of a band
assignable for the nitrile group at 2222 cmꢀ1 region confirm-
ing the cyclized form structure (Scheme 1).
Reaction of 2-pyrrolidinylidenepropanedinitrile 1 with chloro-
acetanilides 2a,b in refluxing acetone in the presence of
anhydrous potassium carbonate, afforded directly 6-amino-
7-cyano-2,3-dihydro-N-(2-substituted phenyl)-1H-pyrrolizine-5-
carboxamides 4a,b. The structure of 4 was established
through spectroscopic (IR, 1H-NMR) as well as elemental
analyses data. The IR spectra of the products reveal one nitrile
stretching vibration band at n ¼ 2221–2220 cmꢀ1 region
and the absence of a second band assignable for the second
nitrile group which confirmed the cyclized structure, in
addition to the appearance of NH2 bands at 3425 and
3439 cmꢀ1. 1H-NMR spectra exhibit the amino group protons
as a singlet signal at d ¼ 3.49–3.86 region (disappeared upon
deuteration). The reaction was assumed to take place via
dehydrohalogenation under the effect of basic potassium
carbonate used in the reaction with subsequent cyclization
due to nucleophilic attack of the active acetanilide methyl-
ene function at one of the nitrile groups giving eventually 4.
The absence of the methylene acetanilide protons in 1H-NMR
spectra, excluded the presence of the open-chain intermedi-
ate 3.
Anti-inflammtory activity screening
The anti-inflammatory activity of the 14 representative com-
pounds 4a–5b, 8a–9a, 9c–10b, was determined in vivo by the
acute carrageenan-induced paw edema standard method in
rats [21–23]. From the obtained results (Table 1), it has been
observed that several newly prepared compounds (4a, 4b,
10a, and 10b) reveal better anti-inflammatory properties
(28.8–36.1% inhibition of edema) comparable to that of diclo-
fenac which was used as a reference standard (28.4% inhi-
bition of edema).
Structure—activity relationships based on the observed
results indicated that, the type of aryl group substitution
attached to the N-position of carboxamide residue plays a
controlling role for developing the exhibited pharmacologi-
cal properties. It has been noticed that, substitution of the
Acylation of 4a,b with acetyl chloride in dry benzene
afforded the mono-acetylated derivatives 5a,b which struc-
tures were deduced by 1H-NMR spectra exhibiting the acetyl
protons as singlet signals at d ¼ 2.10–2.29 region.
Meanwhile, the reaction of 4a,b with aromatic aldehydes
(benzaldehyde and p-anisaldehyde) in refluxing ethanol in
the presence of a catalytic amount of glacial acetic acid gave
the corresponding Schiff bases 6a–c and their structures were
confirmed on the basis of their elemental and spectral data.
The IR spectra of the products reveal the disappearance of the
phenyl group with an electron-withdrawing group,
a
chlorine atom, seems more favorable for constructing an
anti-inflammatory active agent than the case of substitution
with an electron-donating group, a methyl residue, as exhib-
ited in pairs 4a,b (36.1, 31.6% inhibition of edema), 5a,b (23.9,
23.2% inhibition of edema), 8a,c (22.2, 20.3% inhibition of
edema), 8b,d (18.0, 17.5% inhibition of edema), 9a,d (7.1, 3.9%
inhibition of edema), 9c,e (5.7, 3.2% inhibition of edema) and
10a,b (30.3, 28.8% inhibition of edema, respectively).
NH2 bands at 3425 and 3439 cmꢀ1 and H-NMR spectra dis-
1
play the azomethine proton as a sharp singlet signal at
d ¼ 9.08–9.19 region.
However, comparing the activity of 4a and 4b with their
acetylated products 5a, 5b, 8a–d, it was observed that by
blocking the amino group by acetylation, the anti-inflamma-
tory activity decreased and decreased more by nucleophilic
substitution of the acetylated compounds 8a–d with an ali-
cyclic-amino residue (morpholinyl residue) as exhibited in
compounds 9a, 9c–9e. On the other hand, comparing the
activity of the acetylated compounds 5a, 5b, 8a–d, it was
On the other hand, reaction of 4a,b with chloroacetyl
chloride 7a yielded the chloroacetylamino-1H-pyrrolizine
derivatives 8a,c and their structures were confirmed on
the basis of their elemental and spectral data. The latter upon
reaction with secondary amines ‘‘namely morpholine and 1-
methylpiperazine’’ in refluxing ethanol in the presence of
sodium bicarbonate underwent aromatic nucleophilic
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58
F. F. Barsoum
Arch. Pharm. Chem. Life Sci. 2011, 1, 56–65
R
CN
CN
CH2CONH
CN
CN
ClCH2CONH
2
(CH3)2CO, anhy. K2CO3
N
NH
1
R
3
CN
CN
NHCOCH3
CH3COCl
C6H6
N
NH2
OHC
R`
CONH
N
C2H5OH, CH3COOH
5
CONH
R
4
CN
R
90% HCOOH
R`
N=CH
CONH
N
Cl(CH2)nCOCl 7
C6H6
O
6
H
N
R
CN
N
N
NHCO(CH2)nCl
HN
X
N
CONH
10
C2H5OH, NaHCO3
CONH
R
CN
8
R
N
NHCO(CH2)n
CONH
X
N
9
R
2a, 4a; 5a; R = Cl
2b, 4b; 5b; R = CH3
6a; R = Cl, R` = H
7b; n = 2
9b; R = Cl, n = 1, X = NCH3
9c; R = Cl, n = 2, X = O
9d; R = CH3, n = 1, X = O
9e; R = CH3, n = 2, X = O
10a; R = Cl
8a; R = Cl, n = 1
8b; R = Cl, n = 2
6b; R = Cl, R` = OCH3
6c; R = CH3, R` = H
7a; n = 1
8c; R = CH3, n = 1
8d; R = CH3, n = 2
9a; R = Cl, n = 1, X = O
10b; R = CH3
Scheme 1. Proposed synthesis routes.
observed that replacing the hydrogen atom in compounds 5a
and 5b by a chlorine atom in compounds 8a and 8c reduces
the anti-inflammatory activity as exhibited in compounds 5a,
8a (23.9, 22.2% inhibition of edema) and 5b, 8c (23.2, 20.3%
inhibition of edema). In addition, comparing the activity of
the acetylated compounds 8a–d, it was observed that increas-
ing the number of carbon atoms from 1 to 2 of the side chain
at position 6, decreased the anti-inflammatory activity as
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Arch. Pharm. Chem. Life Sci. 2011, 1, 56–65
Anti-inflammatory 1H-Pyrrolizine-5-carboxamides
59
Table 1. Anti-inflammatory activity of the tested compounds using
acute carrageenan-induced paw edema in rats at concentration
20 mg/kg body weight.
compounds to the tricyclic pyrimido[5,4-a]pyrrolizine-9-car-
boxamides derivatives 10a and 10b. However, a decrease in
the anti-inflammatory activity was recognized by the substi-
tution of the primary amino group of the bicyclic pyrrolizine-
5-carboxamides and this activity decreased more by increas-
ing the number of carbon atoms substituted to the amino
group 8b, 8d, 9c, and 9e.
Compound
Mean swelling
volume (mL)
% Inhibition
of edema
Control
Diclofenac sodium
2.583 ꢁ 0.095b
1.850 ꢁ 0.239a
1.650 ꢁ 0.067a
1.767 ꢁ 0.152a
1.967 ꢁ 0.156a
1.983 ꢁ 0.079a
2.010 ꢁ 0.144a
2.117 ꢁ 0.114a
2.060 ꢁ 0.206a
2.130 ꢁ 0.092a
2.401 ꢁ 0.151b
2.435 ꢁ 0.115b
2.483 ꢁ 0.060b
2.500 ꢁ 0.068b
1.800 ꢁ 0.113a
1.840 ꢁ 0.062a
00.0
28.4
36.1
31.6
23.9
23.2
22.2
18.0
20.3
17.5
7.1
5.7
3.9
3.2
30.3
28.8
4a
4b
5a
5b
8a
8b
8c
8d
9a
9c
Ulcerogenic liability
Ulcerogenic liability of the most promising prepared anti-
inflammatory active agents (4a, 4b, 10a, and 10b) was deter-
mined following the previously reported standard method
[24–26] using diclofenac sodium as a reference standard.
From the obtained data (Table 2) it has been noticed that,
compounds 4a and 4b reveal the lowest ulcer indexes (9.2,
11.3, respectively) and they are considered more safer than
diclofenac sodium (reference standard) itself which reveals
ulcer index 13.3.
9d
9e
10a
10b
a
Molecular modeling studies
Statistically significant from the control at p < 0.05.
Statistically significant from diclofenac sodium at p < 0.05.
b
Molecular modeling studies were initiated herein in order to
validate the attained pharmacological data and provide
understandable evidence for the observed anti-inflammatory
behavior. Docking studies were performed by Molecular
Operating Environment (MOE, Version 2005.06, Chemical
Computing Group Inc., Montreal, Quebec, Canada) using
IPXX file downloaded from Protein Data Bank, exhibiting
COX-2 enzyme co-crystallized with diclofenac (the used refer-
ence standard in the anti-inflammatory activity screening
study), which used as a template in the present study. 100
Docking interactions for each ligand were performed and the
top score docking energy value was recorded (Table 3).
Diclofenac was docked in the active site of COX-2 enzyme
using force-field energy MMFX9 with S ¼ ꢀ7.5072 kcal/mol,
exhibiting interaction with two amino acids of the COX-2
active site, which are Ser-530 and Tyr-385. Ser-530 exhibits
two hydrogen bonding interactions of its amino function
exhibited in compounds 8a, 8b (22.2, 18.0% inhibition of
edema) and 8c, 8d (20.3, 17.5% inhibition of edema) and
the same observation was also recognized by nucleophilic
substitution of these compounds with a morpholinyl residue
as exhibited in compounds 9a, 9c (7.1, 5.7% inhibition of
edema) and 9d, 9e (3.9, 3.2% inhibition of edema). However,
upon cyclization of the most potent compounds 4a and 4b
using formic acid, the obtained tricyclic compounds 10a and
10b showed slight decrease in the anti-inflammatory activity
(30.3, 28.8% inhibition of edema) compared to 4a and 4b and
a slight increase in the activity when compared to diclofenac,
reference standard (28.4% inhibition of edema).
˚
As a conclusion, it was found that the bicyclic pyrrolizine-5-
carboxamides 4a and 4b with a primary amino group at
position 6 show the highest anti-inflammatory activity and
this activity was also retained by cyclization of these
with the carboxylic oxygens of diclofenac (d ¼ 2.7, 3.0 A).
However, Tyr-385 exhibits only one hydrogen bond due its
amino acid interaction with the carbonyl carboxylate of
˚
diclofenac (d ¼ 2.7 A).
Table 2. Ulcerogenic liability of the most promising prepared anti-inflammatory active agents.
Compound
Number of
animals with ulcer
% incidence
divided by 10
Average of
ulcer number
Average
severity
Ulcer
index
Control
Diclofenac sodium
4a
4b
10a
10b
0/6
5/6
4/6
5/6
5/6
6/6
0.0
8.3
6.7
8.3
8.3
10.0
0.0
3.7
1.7
1.8
4.8
5.2
0.0
1.3
0.8
1.2
2.0
2.2
0.0
13.3
9.2
11.3
15.1
17.4
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F. F. Barsoum
Arch. Pharm. Chem. Life Sci. 2011, 1, 56–65
˚
Table 3. Docking results for the highly effective anti-inflammatory
agents.
(d ¼ 2.7 A). Meanwhile, docking 4b in the active site of COX-2,
reveals docking energy score S ¼ ꢀ8.856 kcal/mol providing
only one hydrogen bonding due to interaction of amino
Compound
Docking energy
score (kcal/mol)
Amino
acid
Hydrogen
˚
bond (A)
˚
group of 4b and carboxylic residue of Ser-530 (d ¼ 2.7 A)
(Fig. 1). Meanwhile, docking of either compound 10a or
10b (Fig. 2) in the active site of COX-2 reveals only one hydro-
gen bonding due to interaction of N-1 of the pyrimido[5,4-
a]pyrrolizines with the carboxylic residue of Ser-530 (d ¼ 2.8,
Diclofenac sodium
S7.5072
Ser-530
Tyr-385
Tyr-355
Arg-120
Ser-530
Ser-530
Ser-530
2.7, 3.0
2.7
3.1
2.7
2.7
4a
ꢀ10.1032
˚
4b
10a
10b
ꢀ8.856
2.7 A, docking score values S ¼ ꢀ5.4431, ꢀ3,0544,
ꢀ5.4431
ꢀ3.0544
2.8
2.7
respectively).
Alternatively, none of the prepared compounds exhibit any
interaction with COX-1 enzyme. Based on the above obser-
vations, it could be concluded that the highly observed anti-
inflammatory active results (4a,b, 10a,b) are attributed due to
inhibitory properties of these compounds to COX-2 enzyme
participated in prostaglandins generations in the living body.
As a conclusion, the molecular modeling studies revealed
the importance of the primary amino group in the bicyclic
pyrrolizine-5-carboxamides. This primary amino group inter-
Docking of the highly observed anti-inflammatory active
agents 4a,b; 10a,b were studied using the same mentioned
procedure. Docking compound 4a in the active site of COX-2
revealed docking energy score S ¼ ꢀ10.1032 kcal/mol, exhib-
iting two distinguished interactions with two different
amino acids. The first observed one is due to interaction of
amino function of 4a with Tyr-355 carboxylic oxygen
˚
acted with the amino acid Tyr-355 by a hydrogen bond 3.1 A
˚
˚
in compound 4a and with Ser-530 by a hydrogen bond 2.7 A
(d ¼ 3.1 A). The other observed one is due to Arg-120 amino
function interaction with carbonitrile nitrogen of 4a
in compound 4b. However, upon substitution of this primary
Figure 1. Docking of 4b in the active site of COX-2 (S ¼ ꢀ8.1856 kcal/mol) exhibiting interaction with one amino acid, Ser-530 with one
˚
hydrogen bond 2.7 A.
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Arch. Pharm. Chem. Life Sci. 2011, 1, 56–65
Anti-inflammatory 1H-Pyrrolizine-5-carboxamides
61
Figure 2. Docking of 10b in the active site of COX-2 (S ¼ ꢀ3.0544 kcal/mol) exhibiting interaction with one amino acid, Ser-530 with one
˚
hydrogen bond 2.7 A.
amino group to obtain compounds with secondary amino
group, the biological screening showed a marked decrease in
the activity to the extent that they were not detected by the
molecular modeling. On the other hand, converting this
primary amino group to a tertiary one by cyclization of 4a
and 4b, a slight decrease in the activity was recognized in the
biological screening and confirmed in the molecular model-
ing studies by interacting with the amino acid Ser-530 by a
Synthesis of 6-amino-7-cyano-2,3-dihydro-N-(2-
substituted phenyl)-1H-pyrrolizine-5-carboxamide
4a,b
A mixture of equimolar amounts of 2-pyrrolidinylidenepropane-
dinitrile
1 and the corresponding chloroacetanilides 2a,b
(10 mmol) in dry acetone (20 mL) containing anhydrous potas-
sium carbonate (20 mmol), was boiled under reflux for 24 h. The
reaction mixture was filtered while hot and the clear solution
was evaporated till dryness under reduced pressure. The remain-
ing residue was crystallized from a suitable solvent affording the
corresponding 4a,b.
˚
hydrogen bond 2.8 A for compound 10a and 2.7 A for 10b.
˚
Experimental
6-Amino-N-(2-chlorophenyl)-7-cyano-2,3-dihydro-1H-
pyrrolizine-5-carboxamide 4a
Melting points are uncorrected and recorded on an
Electrothermal 9100 digital melting point apparatus. IR spectra
(KBr) were recorded on a Bruker Vector 22 and Jasco FT/IR plus
460 spectrophotometers. 1H-NMR spectra were recorded on a
Varian MERCURY 300 (300 MHz) spectrometer. The starting com-
pounds 1 [27, 28] and 2a,b [29] were prepared according to the
previously reported procedures.
Colorless crystals from ethanol, mp 176–1788C, yield 85%.
IR: nmax cmꢀ1 3425, 3318 (NH, NH ), 2221 (C N), 1636 (C O),
ꢀ
–
–
ꢀ
ꢀ
2
1600, 1586 (C C). 1H-NMR (CDCl3): d 2.54 (pentat, 2H, pyrrol.
–
–
H-2, J ¼ 7.5 Hz), 2.96 (t, 2H, pyrrol. H-1, J ¼ 7.5 Hz), 3.86 (br.s,
2H, D2O exchangeable NH2), 4.37 (t, 2H, pyrrol. H-3, J ¼ 7.2 Hz),
7.00–8.45 (m, 4H, arom. H), 9.27 (br.s, 1H, D2O exchangeable
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62
F. F. Barsoum
Arch. Pharm. Chem. Life Sci. 2011, 1, 56–65
NH). Anal. calcd. for C15H13ClN4O (300.73): C, 59.90; H, 4.36; N,
18.63. Found: C, 60.19; H, 4.29; N, 18.66.
(t, 2H, pyrrol. H-1, J ¼ 7.5 Hz), 4.55 (t, 2H, pyrrol. H-3, J ¼ 7.2 Hz),
–
7.07–8.30 (m, 9H, arom. H), 9.18 (s, 1H, CH N), 10.38 (br.s,
for
–
1H,
D2O
exchangeable
NH).
Anal.
calcd.
C22H17ClN4O (388.84): C, 67.95; H, 4.41; N, 14.41. Found: C,
68.29; H, 4.33; N, 14.63.
6-Amino-7-cyano-2,3-dihydro-N-(2-methylphenyl)-1H-
pyrrolizine-5-carboxamide 4b
Colorless crystals from methanol, mp 165–1678C, yield 87%. IR:
n
max cmꢀ1 3439, 3391, 3323 (NH, NH ), 2220 (C N), 1634 (C O),
N-(2-Chlorophenyl)-7-cyano-2,3-dihydro-6-[[(4-
methoxyphenyl)methylene]amino]-1H-pyrrolizine-5-
carboxamide 6b
ꢀ
–
ꢀ
ꢀ
–
2
1605, 1586 (C C). 1H-NMR (CDCl3): d 2.32 (s, 3H, CH3), 2.51
–
–
(pentat, 2H, pyrrol. H-2, J ¼ 7.2 Hz), 2.96 (t, 2H, pyrrol. H-1,
J ¼ 7.2 Hz), 3.49 (br.s, 2H, D2O exchangeable NH2), 4.36 (t, 2H,
pyrrol. H-3, J ¼ 7.2 Hz), 7.01–8.05 (m, 4H, arom. H), 9.28 (br.s,
1H, D2O exchangeable NH). Anal. calcd. for C16H16N4O (280.32): C,
68.55; H, 5.75; N, 19.99. Found: C, 68.86; H, 5.82; N, 19.83.
Yellow crystals from ethanol, mp 208–2108C, yield 95%. IR:
nmax cmꢀ1 3226 (NH), 2215 (C N), 1665 (C O), 1594, 1539
ꢀ
–
ꢀ
ꢀ
–
(C N, C C). 1H-NMR (CDCl3): d 2.57 (pentat, 2H, pyrrol. H-2,
–
–
–
–
J ¼ 7.5 Hz), 3.07 (t, 2H, pyrrol. H-1, J ¼ 7.5 Hz), 3.90 (s, 3H,
OCH3), 4.54 (t, 2H, pyrrol. H-3, J ¼ 7.2 Hz), 6.98–8.30 (m, 8H,
–
arom. H), 9.08 (s, 1H, CH N), 10.43 (br.s, 1H, D O exchangeable
–
NH). Anal. calcd. for C23H19ClN4O2 (418.86): C, 65.95; H, 4.57; N,
Reaction of 4a,b with acetyl chloride
2
A solution of the appropriate 4 (2.5 mmol) in dry benzene
(10 mL) containing acetyl chloride (5 mmol) was stirred at room
temperature (25–308C) for 48 h. The solid separated upon evap-
orating the reaction mixture till dryness under reduced pressure
was collected, washed with water and crystallized from a suitable
solvent affording the corresponding 5a,b.
13.38. Found: C, 66.28; H, 4.41; N, 13.30.
7-Cyano-2,3-dihydro- N-(2-methylphenyl)-6-
[(phenylmethylene)amino]-1H-pyrrolizine-5-carboxamide
6c
Yellow crystals from ethanol, mp 169–1718C, yield 89%. IR: nmax
cmꢀ1 3290 (NH), 2209 (C N), 1661 (C O), 1592, 1536 (C N, C C).
6-Acetylamino-N-(2-chlorophenyl)-7-cyano-2,3-dihydro-
1H-pyrrolizine-5-carboxamide 5a
ꢀ
–
–
–
ꢀ
ꢀ
–
–
–
1H-NMR (CDCl3): d 2.25 (s, 3H, CH3), 2.57 (pentat, 2H, pyrrol. H-2,
J ¼ 7.8 Hz), 3.08 (t, 2H, pyrrol. H-1, J ¼ 7.5 Hz), 4.56 (t, 2H, pyrrol.
Colorless crystals from ethanol/chloroform as 2:1 v/v, mp 222–
2248C, yield 81%. IR: nmax cmꢀ1 3378, 3208 (NH), 2226 (C N),
ꢀ
–
–
ꢀ
H-3, J ¼ 7.2 Hz), 7.10–7.89 (m, 9H, arom. H), 9.19 (s, 1H, CH N),
ꢀ
1
–
–
1671 (C O), 1591, 1561 (C C). H-NMR (CDCl ): d 2.29 (s, 3H, CH ),
9.99 (br.s, 1H, D2O exchangeable NH). Anal. calcd.
for C23H20N4O (368.42): C, 74.98; H, 5.47; N, 15.21. Found: C,
75.23; H, 5.45; N, 15.13.
–
–
3
3
2.56 (pentat, 2H, pyrrol. H-2, J ¼ 6.6 Hz), 3.00 (t, 2H, pyrrol. H-1,
J ¼ 6.9 Hz), 4.42 (t, 2H, pyrrol. H-3, J ¼ 6.3 Hz), 7.04–8.35 (m, 5H,
arom. H þ NH), 8.78 (br.s, 1H, D2O exchangeable NH). Anal. calcd.
for C17H15ClN4O2 (342.77): C, 59.56; H, 4.41; N, 16.35. Found: C,
59.90; H, 4.41; N, 16.51.
Reaction of 4a,b with chloroacetyl chloride and 3-
chloropropionyl chloride 7a,b
A mixture of the appropriate 4 (2 mmol) and the corresponding
7a,b (4 mmol) in dry benzene (10 mL) was stirred at room
temperature (25–308C) for 48 h. The separated solid was col-
lected, washed with water and crystallized from a suitable sol-
vent affording the corresponding 8a–d.
6-Acetylamino-7-cyano-2,3-dihydro-N-(2-methylphenyl)-
1H-pyrrolizine-5-carboxamide 5b
Colorless crystals from methanol, mp 208–2108C, yield 79%. IR:
n
max cmꢀ1 3416, 3267 (NH), 2224 (C N), 1667 (C O), 1588, 1540
–
–
ꢀ
–
ꢀ
ꢀ
–
(C C). 1H-NMR (DMSO-d6): d 2.10 (s, 3H, CH3), 2.22 (s, 3H, CH3),
2.43–2.52 (m, 2H, pyrrol. H-2), 2.99 (t, 2H, pyrrol. H-1, J ¼ 7.2 Hz),
4.92 (t, 2H, pyrrol. H-3, J ¼ 7.5 Hz), 7.07–7.69 (m, 4H, arom. H),
8.84 (br.s, 1H, D2O exchangeable NH), 9.96 (br.s,
1H, D2O exchangeable NH). Anal. calcd. for C18H18N4O2
(322.35): C, 67.06; H, 5.63; N, 17.38. Found: C, 66.98; H, 5.85;
N, 17.58.
6-[(Chloroacetyl)amino]-N-(2-chlorophenyl)-7-cyano-2,3-
dihydro-1H-pyrrolizine-5-carboxamide 8a
Colorless crystals from ethanol/chloroform as 3:1 v/v, mp 217–
2198C, yield 76%. IR: nmax cmꢀ1 3385, 3212 (NH), 2222 (C N),
ꢀ
ꢀ
ꢀ
1
–
–
1667 (C O), 1592, 1568 (C C). H-NMR (CDCl ): d 2.59 (pentat, 2H,
–
–
3
pyrrol. H-2, J ¼ 7.5 Hz), 3.06 (t, 2H, pyrrol. H-1, J ¼ 7.5 Hz), 4.31 (s,
2H, CH2Cl), 4.47 (t, 2H, pyrrol. H-3, J ¼ 7.2 Hz), 7.06–8.39 (m, 4H,
arom. H), 8.50 (br.s, 1H, D2O exchangeable NH), 8.66 (br.s,
1H, D2O exchangeable NH). Anal. calcd. for C17H14Cl2N4O2
(377.22): C, 54.12; H, 3.74; N, 14.85. Found: C, 54.29; H, 3.67;
N, 14.81.
Reaction of 4a,b with aromatic aldehydes
A mixture of equimolar amounts of the appropriate 4 and the
corresponding aromatic aldehyde (2 mmol) in absolute ethanol
(10 mL) containing glacial acetic acid (3–4 drops) was boiled
under reflux for 4 h. The separated solid was collected and
crystallized from a suitable solvent affording the corresponding
6a–c.
6-[(Chloropropionyl)amino]-N-(2-chlorophenyl)-7-cyano-
2,3-dihydro-1H-pyrrolizine-5-carboxamide 8b
Colorless crystals from methanol, mp 218–2208C, yield 73%. IR:
N-(2-Chlorophenyl)-7-cyano-2,3-dihydro-6-
[(phenylmethylene)amino]-1H-pyrrolizine-5-carboxamide
n
max cmꢀ1 3376, 3225 (NH), 2223 (C N), 1669 (C O), 1592, 1563
ꢀ
–
ꢀ
ꢀ
–
1
–
–
(C C). H-NMR (CDCl ): d 2.57 (pentat, 2H, pyrrol. H-2, J ¼ 7.5 Hz),
3
6a
Yellow crystals from benzene, mp 206–2088C, yield 91%. IR: nmax
2.94–3.06 (m, 4H, pyrrol. H-1 þ CH2Cl), 3.88 (t, 2H, COCH2,
J ¼ 6.6 Hz), 4.43 (t, 2H, pyrrol. H-3, J ¼ 7.2 Hz), 7.05–8.65 (m,
6H, arom. H þ 2 NH). Anal. calcd. for C18H16Cl2N4O2 (391.25):
C, 55.25; H, 4.12; N, 14.32. Found: C, 55.61; H, 3.98; N, 14.30.
cmꢀ1 3227 (NH), 2212 (C N), 1670 (C O), 1611, 1590 (C N, C C).
ꢀ
–
–
–
ꢀ
ꢀ
–
–
–
1H-NMR (CDCl3): d 2.58 (pentat, 2H, pyrrol. H-2, J ¼ 7.5 Hz), 3.08
ß 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
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Arch. Pharm. Chem. Life Sci. 2011, 1, 56–65
Anti-inflammatory 1H-Pyrrolizine-5-carboxamides
63
6-[(Chloroacetyl)amino]-7-cyano-2,3-dihydro-N-(2-
methylphenyl)-1H-pyrrolizine-5-carboxamide 8c
N-(2-Chlorophenyl)-7-cyano-2,3-dihydro-6-[[3-(4-
morpholinyl)propionyl]amino]-1H-pyrrolizine-5-
carboxamide 9c
Colorless crystals from methanol, mp 184–1868C, yield 74%. IR:
n
max cmꢀ1 3429, 3259 (NH), 2224 (C N), 1671, 1614 (C O), 1587,
ꢀ
–
ꢀ
ꢀ
–
Colorless crystals from ethanol, mp 188–1908C, yield 65%. IR:
1
n
max cmꢀ1 3378, 3231 (NH), 2226 (C N), 1666 (C O), 1591, 1562
–
1527 (C C). H-NMR (CDCl ): d 2.29 (s, 3H, CH ), 2.56 (pentat, 2H,
ꢀ
–
–
3
3
ꢀ
ꢀ
–
1
pyrrol. H-2, J ¼ 7.5 Hz), 3.04 (t, 2H, pyrrol. H-1, J ¼ 7.5 Hz), 4.26 (s,
2H, CH2Cl), 4.40 (t, 2H, pyrrol. H-3, J ¼ 7.5 Hz), 7.11–7.27 (m, 4H,
arom. H), 8.37 (br.s, 1H, D2O exchangeable NH), 8.50 (br.s,
1H, D2O exchangeable NH). Anal. calcd. for C18H17ClN4O2
(356.80): C, 60.59; H, 4.80; N, 15.70. Found: C, 60.79; H, 4.69;
N, 15.83.
–
(C C). H-NMR (CDCl ): d 2.55 (pentat, 2H, pyrrol. H-2, J ¼ 7.5 Hz),
–
3
2.67–2.78 (m, 6H, morpholinyl 2 NCH2 þ COCH2CH2), 2.91 (t, 2H,
COCH2CH2, J ¼ 5.1 Hz), 3.03 (t, 2H, pyrrol. H-1, J ¼ 7.5 Hz), 3.80
(t, 4H, morpholinyl 2 OCH2, J ¼ 3.9 Hz), 4.45 (t, 2H, pyrrol. H-3,
J ¼ 7.5 Hz), 7.05–8.29 (m, 4H, arom. H), 9.03 (s, 1H, D2O exchange-
able NH), 10.98 (s, 1H, D2O exchangeable NH). Anal. calcd.
for C22H24ClN5O3 (441.90): C, 59.79; H, 5.47; N, 15.85. Found:
C, 60.07; H, 5.80; N, 15.99.
6-[(Chloropropionyl)amino]-7-cyano-2,3-dihydro-N-(2-
methylphenyl)-1H-pyrrolizine-5-carboxamide 8d
7-Cyano-2,3-dihydro-N-(2-methylphenyl)-6-[(4-
morpholinylacetyl)amino]-1H-pyrrolizine-5-carboxamide
Colorless crystals from methanol, mp 198–1998C, yield 68%. IR:
n
max cmꢀ1 3323, 3289 (NH), 2221 (C N), 1704, 1648 (C O), 1528,
ꢀ
–
ꢀ
ꢀ
–
1
–
–
1490 (C C). H-NMR (CDCl ): d 2.29 (s, 3H, CH ), 2.54 (pentat, 2H,
3
3
9d
Colorless crystals from methanol, mp 219–2218C, yield 75%. IR:
pyrrol. H-2, J ¼ 7.5 Hz), 2.90–3.03 (m, 4H, pyrrol. H-1 þ CH2Cl),
3.82 (t, 2H, COCH2, J ¼ 6.3 Hz), 4.37 (t, 2H, pyrrol. H-3, J ¼ 7.2 Hz),
7.09–7.62 (m, 4H, arom. H), 8.04 (br.s, 1H, D2O exchangeable NH),
8.64 (br.s, 1H, D2O exchangeable NH). Anal. calcd.
for C19H19ClN4O2 (370.82): C, 61.54; H, 5.16; N, 15.11. Found:
C, 61.39; H, 5.13; N, 14.97.
n
max cmꢀ1 3253, 3215 (NH), 2221 (C N), 1651 (C O), 1598, 1566
ꢀ
–
ꢀ
ꢀ
–
1
–
(C C). H-NMR (CDCl ): d 2.14 (s, 3H, CH ), 2.65 (pentat, 2H, pyrrol.
–
3
3
H-2, J ¼ 7.5 Hz), 2.70 (t, 4H, morpholinyl 2 NCH2, J ¼ 4.8 Hz),
3.15 (t, 2H, pyrrol. H-1, J ¼ 7.5 Hz), 3.55 (s, 2H, COCH2), 3.79
(t, 4H, morpholinyl 2OCH2, J ¼ 4.5 Hz), 4.42 (t, 2H, pyrrol. H-3,
J ¼ 7.5 Hz), 7.18–7.38 (m, 4H, arom. H), 9.10 (br.s, 1H, D2O
exchangeable NH), 9.38 (br.s, 1H, D2O exchangeable NH). Anal.
calcd. for C22H25N5O3 (407.46): C, 64.85; H, 6.18; N, 17.19. Found:
C, 64.60; H, 5.99; N, 17.16.
Reaction of 8a-d with secondary amines
A mixture of the appropriate 8a–d (2 mmol) and the correspond-
ing secondary amine (4 mmol) in absolute ethanol (10 mL) con-
taining sodium bicarbonate (4 mmol), was boiled under reflux
for 8 h. The reaction mixture was filtered while hot. The solid
separated upon storing the clear reaction mixture at room
temperature overnight, was collected and crystallized from a
suitable solvent affording the corresponding 9a–e.
7-Cyano-2,3-dihydro-N-(2-methylphenyl)-6-[[3-(4-
morpholinyl)propionyl]amino]-1H-pyrrolizine-5-
carboxamide 9e
Colorless crystals from methanol, mp 197–1988C, yield 71%.
IR: nmax cmꢀ1 3237 (NH), 2216 (C N), 1649 (C O), 1575, 1536
–
ꢀ
–
ꢀ
ꢀ
–
(C C). 1H-NMR (CDCl3): d 2.31 (s, 3H, CH3), 2.56 (pentat, 2H,
–
N-(2-Chlorophenyl)-7-cyano-2,3-dihydro-6-[(4-
morpholinylacetyl)amino]-1H-pyrrolizine-5-carboxamide
pyrrol. H-2, J ¼ 7.5 Hz), 2.62–2.69 (m, 6H, morpholinyl
2
NCH2 þ COCH2CH2), 2.83 (t, 2H, COCH2CH2, J ¼ 5.4 Hz), 3.02
(t, 2H, pyrrol. H-1, J ¼ 7.5 Hz), 3.79 (t, 4H, morpholinyl 2
OCH2, J ¼ 4.5 Hz), 4.39 (t, 2H, pyrrol. H-3, J ¼ 7.2 Hz), 7.09–
7.72 (m, 4H, arom. H), 9.20 (s, 1H, D2O exchangeable NH),
11.18 (s, 1H, D2O exchangeable NH). Anal. calcd.
for C23H27N5O3 (421.49): C, 65.54; H, 6.46; N, 16.62. Found: C,
65.80; H, 6.37; N, 16.70.
9a
Colorless crystals from benzene/ethanol as 1:1 v/v, mp 229–
2318C, yield 70%. IR: nmax cmꢀ1 3354, 3228 (NH), 2221 (C N),
ꢀ
ꢀ
ꢀ
1712, 1669 (C O), 1595, 1535 (C C). 1H-NMR (CDCl3): d 2.56
–
–
–
–
(pentat, 2H, pyrrol. H-2, J ¼ 7.5 Hz), 2.76 (br. s, 4H, morpholinyl
2 NCH2), 3.04 (t, 2H, pyrrol. H-1, J ¼ 7.5 Hz), 3.31 (s, 2H, COCH2),
3.83 (t, 4H, morpholinyl 2 OCH2, J ¼ 4.5 Hz), 4.46 (t, 2H, pyrrol. H-
3, J ¼ 7.2 Hz), 7.05–8.29 (m, 4H, arom. H), 8.89 (br.s,
1H, D2O exchangeable NH), 9.30 (br.s, 1H, D2O exchangeable
NH). Anal. calcd. for C21H22ClN5O3 (427.88): C, 58.94; H, 5.18;
N, 16.37. Found: C, 59.18; H, 5.08; N, 16.24.
Reaction of 4a,b with formic acid
A mixture of 4a,b (2 mmol) and 90% formic acid (4 mmol) was
heated in a water bath for 3 h, cooled and neutralized with 10%
sodium hydroxide. The separated solid was filtered, washed with
water and crystallized from a suitable solvent affording the
corresponding 10a,b.
N-(2-Chlorophenyl)-7-cyano-2,3-dihydro-6-[(1-methyl-4-
piperazinylacetyl)amino]-1H-pyrrolizine-5-carboxamide 9b
Colorless crystals from methanol, mp 190–1928C, yield 77%. IR:
n
max cmꢀ1 3347, 3251 (NH), 2227 (C N), 1710, 1670 (C O), 1591,
ꢀ
N-(2-Chlorophenyl)-4-oxo-4,5,6,7-tetrahydro-3H-
pyrimido[5,4-a]pyrrolizine-9-carboxamide 10a
–
ꢀ
ꢀ
–
1537 (C C). 1H-NMR (CDCl3): d 2.37 (s, 3H, NCH3), 2.43–2.61 (m,
–
–
6H, pyrrol. H-2 þ piperazinyl 2 NCH2), 2.80 (br.s, 4H, piperazinyl
2 NCH2), 3.04 (t, 2H, pyrrol. H-1, J ¼ 7.2 Hz), 3.29 (s, 2H, COCH2),
4.45 (t, 2H, pyrrol. H-3, J ¼ 7.2 Hz), 7.05–8.28 (m, 4H, arom. H),
8.94 (br.s, 1H, D2O exchangeable NH), 9.25 (br.s,
1H, D2O exchangeable NH). Anal. calcd. for C22H25ClN6O2
(440.92): C, 59.92; H, 5.72; N, 19.06. Found: C, 59.89; H, 5.62;
N, 19.01.
Colorless crystals from benzene, mp 293–2958C, yield 83%. IR:
max cmꢀ1 3378, 3277 (NH), 1699, 1663 (C O), 1594, 1536 (C N,
–
–
–
n
–
C C). 1H-NMR (DMSO-d6): d 2.52 (pentat, 2H, CH2-6, J ¼ 7.8 Hz),
–
–
3.14 (t, 2H, CH2-5, J ¼ 7.5 Hz), 4.25 (t, 2H, CH2-7, J ¼ 7.2 Hz), 7.31–
8.20 (m, 7H, arom. H þ 2 NH þ N ¼ CH). Anal. calcd.
for C16H13ClN4O2 (328.74): C, 58.45; H, 3.98; N, 17.04. Found:
C, 58.51; H, 3.93; N, 17.13.
ß 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

64
F. F. Barsoum
Arch. Pharm. Chem. Life Sci. 2011, 1, 56–65
(Table 2) and the degree of ulcerogenic effect was expressed in
terms of:
N-(2-Methylphenyl)-4-oxo-4,5,6,7-tetrahydro-3H-
pyrimido[5,4-a]pyrrolizine-9-carboxamide 10b
Colorless crystals from benzene/methanol as 2:1 v/v, mp 278–
2808C, yield 81%. IR: nmax cmꢀ1 3421 (NH), 1689, 1658 (C O),
1. Percentage incidence of ulcer divided by 10.
2. Average number of ulcers per stomach.
3. Average severity of ulcers.
–
–
1601, 1534 (C N, C C). 1H-NMR (DMSO-d6): d 2.24 (s, 3H, CH3),
–
–
–
–
2.36–2.54 (m, 2H, CH2-6), 3.14 (t, 2H, CH2-5, J ¼ 7.8 Hz), 4.19 (t,
–
2H, CH -7, J ¼ 7.8 Hz), 7.20–7.49 (m, 5H, arom. H þ N CH), 8.33
–
2
The ulcer index is the value that resulted from the sum of the
above three values.
(s, 1H, D2O exchangeable NH), 9.75 (s, 1H, D2O exchangeable NH).
Anal. calcd. for C17H16N4O2 (308.33): C, 66.22; H, 5.23; N, 18.17.
Found: C, 66.50; H, 5.02; N, 18.09.
I am grateful to Department of Pharmacology, National Research Centre,
Dokki, Cairo, Egypt, for allowing the performance of pharmacological
screening and for their kind interest and valuable discussions.
Anti-inflammatory activity screening
Anti-inflammatory activity screening for the prepared com-
pounds 4a–5b, 8a–9, 9c–10b was determined in vivo by the acute
carrageenan-induced paw edema standard method in rats
[21–23]. Wister albino rats of either sex (pregnant female animals
were excluded) weighing 160–180 g were divided into 16 groups
of 6 animals each. Administration of diclofenac sodium (refer-
ence standard) and the tested compounds dissolved in saline
solution and 0.2 mL DMSO, at a dose of 20 mg/kg (body weight)
was given intraperitoneally 1 h before induction of inflam-
mation. The control group was given saline solution containing
0.2 mL DMSO. Carrageenan paw edema was induced by subcu-
taneous injection of 1% solution of carrageenan in saline
(0.1 mL/rat) into the right hind paw of rats. Paw volumes were
measured volumetrically after 4 h of inflammation induction
with plethysmometer 7140 (UGO BASILE, Italy) and compared
with the initial hind paw volume of each rat for determining the
oedema volume. Data were collected, checked, revised and ana-
lyzed. Quantitative variables from normal distribution were
expressed as means ꢁ SE ‘‘standard error’’. The significant differ-
ence between groups was tested by using one-way ANOVA fol-
lowed by post hoc test and the chosen level of significance was
p < 0.05.
The author has declared no conflict of interest.
References
[1] L. Ferrero-Miliani, O. H. Nielsen, P. S. Andersen, S. E.
Girardin, Clin. Exp. Immunol. 2007, 147, 227–235.
[2] J. G. Black, Microbiology Principles and Exploration, 6th Ed., John
Wiley and Sons, Inc., Hoboken 2005, pp. 446–467.
[3] K. D. Bosscher, W. V. Berghe, G. Haegeman, J. Neuroimmunol.
2000, 109, 16–22.
[4] M. Stoeck, R. Riedel, G. Hochhaus, D. Ha¨fner, J. M. Masso,
B. Schmidt, A. Hatzelmann, D. Marx, D. S. Bundschuh,
J. Pharmacol. Exp. Ther. 2004, 309, 249–258.
[5] J. Ehrchen, L. Steinmu¨ller, K. Barczyk, K. Tenbrock, W.
Nacken, M. Eisenacher, U. Nordhues, C. Sorg, C.
Sunderko¨tter, J. Roth, Blood 2007, 109, 1265–1274.
[6] A. Z. A. Leite, A. M. Sipahi, A. O. M. C. Damia˜o, A. T. Garcez,
C. A. Buchpiguel, F. P. Lopasso, M. L. L. Lordello, C. L. O.
Agostinho, A. A. Laudanna, Braz. J. Med. Biol. Res. 2004, 37,
333–336.
The anti-inflammatory activity was expressed as percentage
inhibition of edema volume in treated animals in comparison
with the control group (Table 1).
[7] O. Abdul-Hadi, J. Parvizi, M. S. Austin, E. Viscusi, T. Einhorn,
J. Bone Joint Surgery 2009, 91, 2020–2027.
VcꢀVt
[8] F. Bao, S. M. John, Y. Chen, R. D. Mathison, L. C. Weaver,
Neuroscience 2006, 140, 1011–1022.
% Inhibition of edema ¼
ꢂ 100
Vc
[9] R. E. Dery, M. Ulanova, L. Puttagunta, G. R. Stenton, D.
James, S. Merani, R. Mathison, J. Davison, A. D. Befus, Eur.
J. Immunol. 2004, 34, 3315–3325.
Where, Vc and Vt are the volumes of edema for the control and
drug-treated animal groups, respectively.
[10] A. Jain, E. Basal, Phytomedicine 2003, 10, 34–38.
Ulcerogenic liability
[11] R. Grzanna, L. Lindmark, C. G. Frondoza, J. Med. Food 2005, 8,
125–132.
The ulcerogenic liability of the most promising prepared anti-
inflammatory active agents (4a, 4b, 10a, and 10b) was deter-
mined in albino rats following the previously reported standard
method [24–26]. Rats of either sex (pregnant female rats were
excluded) weighing 130–150 g were divided into 6 groups of 6
animals each. The animals were fasted 18 h before drug admin-
istration. Diclofenac sodium (reference standard) and the tested
compounds (at a dose of 20 mg/kg body weight), were suspended
in saline solution by the aid of few drops of Tween 80 and were
administered orally for three successive days to fasted rats. The
control group animals were given saline with few drops of Tween
80. One hour following the last dose, the animals were sacrificed
by cervical dislocation and the stomach was removed, opened
along the greater curvature and rinsed with saline. The gastric
mucosa was examined with a magnifying lens (10ꢂ) for the
presence of lesions and erosions. The ulcer index was calculated
[12] B. J. Chen, Leukemia Lymphoma 2001, 42, 253–265.
[13] M. C. Allison, A. G. Howatson, C. J. Torrance, F. D. Lee, R. I. G.
Russell, N. Engl. J. Med. 1992, 327, 749–754.
[14] M. M. Wolfe, D. R. Lichtenstein, G. Singh, N. Engl. J. Med. 1999,
340, 1888–1899.
[15] M. Y. Ebeid, H. H. Hassanein, N. N. Obidan, A. B. Hassan,
Egypt. J. Pharm. Sci. 1988, 29, 533–543.
[16] M. M. Hanna, M. El-Sayed, Bull. Fac. Pharm. Cairo Univ. 1993, 31,
39–43.
[17] H. H. Hassanein, M. Y. Ebeid, Bull. Fac. Pharm. Cairo Univ. 1993,
31, 33–38.
[18] M. M. Hanna, A. S. Farag, N. M. El-Sayed, M. Y. Ebeid, A. S.
Attia, Az. J. Pharm. Sci. 1994, 14, 133–144.
ß 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

Arch. Pharm. Chem. Life Sci. 2011, 1, 56–65
Anti-inflammatory 1H-Pyrrolizine-5-carboxamides
65
[19] F. A. Ragab, M. M. Hanna, F. F. Barsoum, S. H. Fahim,
M. Y. Ebeid, H. A. Salem, Egypt. J. Chem. 2007, 50, 105–
116.
[24] Y. E. Hamza, O. A. Sammour, H. A. Abdel-Latif, Pharm. Ind.
1994, 56, 286–291.
[25] F. F. Barsoum, H. M. Hosni, A. S. Girgis, Bioorg. Med. Chem.
2006, 14, 3929–3937.
[20] K. M. Amin, M. M. Hanna, H. E. Abo-Youssef, R. F. George, Eur.
J. Med. Chem. 2009, 44, 4572–4584.
[26] F. Barsoum, H. Georgey, N. Abdel-Gawad, Molecules 2009, 14,
667–681.
[21] C. A. Winter, E. A. Risley, G. W. Nuss, Proc. Soc. Exp. Biol. Med.
1962, 111, 544–547.
[27] A. Etienne, Y. Correia, Bull. Soc. Chim. 1969, 10, 3704–3712,
Chem. Abstr., 1970, 72, 55133.
[22] B. Tozkoparan, E. Ku¨peli, E. Yes¸ilada, M. Ertan, Bioorg. Med.
Chem. 2007, 15, 1808–1814.
[28] V. G. Granik, A. M. Zhidkova, R. A. Dubinskii, Khim.
Geterotsikl. Soedin. 1982, 518–522, Chem. Abstr. 1982, 97, 55765.
[23] A. M. Abdel-Megeed, H. M. Abdel-Rahman, G. E. S.
Alkaramany, M. A. El-Gendy, Eur. J. Med. Chem. 2009, 44,
117–123.
[29] M. K. El-Said, S. M. E. Aly, F. A. Romeih, F. F. Barsoum, A. B.
Hassan, Egypt. J. Pharm. Sci. 1991, 32, 251–261.
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