Antitumour activity of 5-[(2E)-1-(1H-benzimidazol-2-yl)-3-substituted phenylprop-2-en-1-ylidene] pyrimidine-2,4,6(1H,3H,5H)-triones against Dalton's ascitic lymphoma in mice

Article abstract of DOI:10.1007/s00044-012-0407-1

A new series of novel benzimidazole derivatives containing barbitone moiety (5a-f) was synthesized by a Knoevenagel condensation of (2E)-1-(1H- benzimidazol-2-yl)-3-phenylprop-2-en-1-ones (4a-f) and barbituric acid in the presence of catalytic amount of acetic acid medium. All the final structures were assigned on the basis of IR, ¹H NMR and mass spectra analysis. Acute toxicity studies were performed initially to determine the safety of titled derivatives and the ED ₅₀ value was calculated 50 mg/kg. All the final derivatives were screened for antitumour activity against Dalton's ascitic lymphoma in mice. All the new candidates at a dose of 50 mg/kg showed a good antitumour activity against DLA-bearing mice when compared to the standard 5-fluro uracil. Among the final derivatives (5e), 5-[(2E)-1-(1H-benzimidazol-2- yl)-3-(3-nitrophenyl) prop-2-en-1-ylidene] pyrimidine-2,4,6(1H,3H,5H)-trione was found to be most potent antitumour in nature.

Full text of DOI:10.1007/s00044-012-0407-1

MEDICINAL
CHEMISTRY
RESEARCH
Med Chem Res
DOI 10.1007/s00044-012-0407-1
ORIGINAL RESEARCH
Antitumour activity of 5-[(2E)-1-(1H-benzimidazol-2-yl)-3-
substituted phenylprop-2-en-1-ylidene] pyrimidine-
2,4,6(1H,3H,5H)-triones against Dalton’s ascitic
lymphoma in mice
•
Bijo Mathew Jerad Suresh Devaraji Vinod
•
Received: 13 June 2012 / Accepted: 6 December 2012
Ó Springer Science+Business Media New York 2012
Abstract A new series of novel benzimidazole deriva-
tives containing barbitone moiety (5a–f) was synthesized by
a Knoevenagel condensation of (2E)-1-(1H-benzimidazol-
2-yl)-3-phenylprop-2-en-1-ones (4a–f) and barbituric acid
in the presence of catalytic amount of acetic acid medium.
All the final structures were assigned on the basis of IR, 1H
NMR and mass spectra analysis. Acute toxicity studies were
performed initially to determine the safety of titled deriv-
atives and the ED50 value was calculated 50 mg/kg. All the
final derivatives were screened for antitumour activity
against Dalton’s ascitic lymphoma in mice. All the new
candidates at a dose of 50 mg/kg showed a good antitumour
activity against DLA-bearing mice when compared to the
standard 5-fluro uracil. Among the final derivatives (5e),
5-[(2E)-1-(1H-benzimidazol-2-yl)-3-(3-nitrophenyl) prop-
2-en-1-ylidene] pyrimidine-2,4,6(1H,3H,5H)-trione was
found to be most potent antitumour in nature.
Introduction
Benzimidazole chalcones are considered as the most
important key intermediate for the development of various
biologically active compounds with some interesting
pharmacological profile such as anticancer, anti-HIV and
anti microbial properties (Shaharyar et al., 2010; Nofal
et al., 2011; Aboul-Fadal et al., 2000; Reddy et al., 2010;
Srivastava et al., 2010). Barbituric acid is pyrimidine-
2,4,6(1H,3H,5H)-trione and its pharmacological action
mainly constituted with the introduction of dialkyl lipo-
philic groups in the 5th position of the activated methylene
group (Shulman et al., 1967). The active methylene group
of barbituric acid condensation with different carbonyl
compounds possess different pharmacological profile such
as antimicrobial, selective cell adhesion inhibitors, anti-
oxidant and DNA cleavage activities have been already
reported (Sangani et al., 2006; Harriman et al., 2008; Bi-
radar et al., 2010).
Keywords Alanine amino transferase ꢀ
Alkaline phosphatase ꢀ Aspartate amino transferase ꢀ
Barbituric acid ꢀ Knoevenagel condensation ꢀ
Total cholesterol
Tumour is a mass of tissues which proliferate rapidly,
spread throughout the body and may eventually cause
death of the host. Chemotherapy is an effective treatment
against various types of cancer either singly or in combi-
nation with surgery or radiotherapy (Mohan, 2006). How-
ever, chemotherapeutic effects of most of the drugs showed
limited efficacies due to the development of various side
effects. This fostered our attempts to evaluate the various
synthetic drugs against cancer as they are less likely to
cause serious side effects. Previously, benzimidazole
derivatives aroused a considerable attention in the antitu-
mour activities (Georges et al., 1994; Badawey et al., 1995;
Cheng et al., 2009). Barbituric acid is an important mem-
ber of the pyrimidine family and is also known to possess a
wide range of antitumour properties (Sing et al., 2009;
Naguib et al., 1993; Jursic et al., 2002). It was envisaged
B. Mathew (&)
Department of Pharmaceutical Chemistry, Grace College
of Pharmacy, Palakkad 678004, Kerala, India
e-mail: bijovilaventgu@gmail.com
J. Suresh ꢀ D. Vinod
Department of Pharmaceutical Chemistry, Madras Medical
College, Chennai 600003, India
123

Med Chem Res
that the above two pharmacophores if linked together by
means of a carbon–carbon double bond would exhibit
novel molecular templates with interesting biological
activities in animal models. So, our research proposal was
the reaction between activated methylene group present in
the barbituric acid with the a, b unsaturated ketone of
benzimidazole derivatives in the presence of a catalytic
amount of acetic acid produced an exocyclic olefinic
linkage.
points were determined using melting point apparatus MP-
DS TID 2,000 V and the values were uncorrected. Reac-
tions were monitored by thin layer chromatography
(methanol:acetone, 6:4) on a pre-coated silica gel G plates
using iodine vapour as visualizing agent. IR spectra were
recorded on JASCO FT/IR-140 spectrophotometer by KBr
pellets technique. PMR spectra were recorded using
BRUCKER FT-NMR-500 MHz spectrophotometer using
DMSO as solvent and TMS as internal standard. The
chemical shift was expressed in d ppm. Mass spectra were
recorded on a JEOL GC mate mass spectrometer.
The synthesis of 5-[(2E)-1-(1H-benzimidazol-2-yl)-3-
phenylprop-2-en-1-ylidene]
substituted
pyrimidine-
2,4,6(1H,3H,5H)-triones (5a–f) described in this study was
outlined in Fig. 1. The chalcones of (4a–f) were prepared
by reacting 2-acetyl benzimidazole with appropriate
substituted aromatic aldehydes in the presence of a base by
Claisen–Schmidt condensation(Reddy et al., 2010; Tale-
sara et al., 2010).The condensation of (2E)-1-(1H-ben-
zimidazol-2-yl)-3-substituted phenylprop-2-en-1-ones with
barbituric acid in presence of acetic acid medium gave the
titled derivatives. The acidic hydrogen present in the
pyrimidine nucleus of barbituric acid undergoes intermo-
lecular acid-catalysed dehydration resulting an activated
C=C bond in the final derivatives (Jursic et al., 2002).
All biochemical investigations were done using COBAS
MIRA PLUS-S Auto analyser from Roche Switzerland.
Haematological tests were carried out in COBAS MICROS
OT 18 from Roche. Newly added Hi-Tech instruments
MAX MAT used for an auto analyser for all biochemistry
investigations in blood sample.
Chemistry
Synthesis of (2E)-1-(1H-benzimidazol-2-yl)-3-substituted
phenylprop-2-en-1-ones (4a–f)
2-Acetyl benzimidazole (0.01 mol) and appropriately
substituted aromatic aldehydes (0.012 mol) were mixed in
ethanol (20 ml) containing 10 % aq. KOH (8 ml) and
magnetically stirred the solution constantly at room tem-
perature for 10 h. The whole mixture was transferred into
100 ml ice cold water and acidified with dil. HCl. The solid
formed was washed, filtered, dried and recrystallized from
absolute ethanol.
Materials and methods
General
All the solvents and chemicals were purchased from
MERCK, Nice chemicals and SD Fine Chemicals. Melting
Fig. 1 Synthetic route of the
titled compounds
O
N
O
N
a
N
H
N
H
CH3
4a-f
R
3a
b
R
H
N
O
O
5a: phenyl
5b: 4- chloro phenyl
5c: 4- methoxy phenyl
5d: 4-N,Ndimethyl aminophenyl
5e: 3- nitro phenyl
NH
N
O
5f: 2-hydroxy phenyl
N
H
R
Scheme 1 Synthesis of (5a-f).Reagents and conditions
a:phenylsubstituted aldehyde,EtOH,10%KOH.10h stirr
b:barbituric acid,acetic acid,7h reflux
123

Med Chem Res
groups’ d is the constant factor between two successive
doses, n is the number of animals in each groups,
the sum of (ZXd).
Synthesis of 5-[(2E)-1-(1H-benzimidazol-2-yl)-3-
substituted phenylprop-2-en-1-ylidene] pyrimidine-
2,4,6(1H,3H,5H)-triones (5a–f)
P
is
Induction of cancer using DLA cells
To a solution of (4a–f) (0.01 mol) in 7 ml acetic acid,
barbituric acid (0.01 mol) was added with constant stirring.
The reaction mixture was then refluxed for 7 h with
occasional stirring. The reaction progress was monitored
by TLC (methanol:acetone, 6:4). The resultant contents
were poured into crushed ice. The crude product was fil-
tered, washed with water, dried and recrystallized from
methanol. The physical characterizations of all the syn-
thesized derivatives are summarized in Table 1.
Male Swiss albino mice (20–25 g) were produced from ani-
mal experimental laboratory and used throughout the study.
Dalton’s lymphoma ascites (DLA) cells were supplied by
Amala cancerresearchcenter, Trissur, Kerala, India. Thecells
maintained in vivo in Swiss albino mice by intraperitoneal
transplantation. While transforming the tumour cells to the
grouped animal, the DLA cells were aspirated from peritoneal
cavity of the mice using saline. The cell counts were done and
further dilutions were made so that total number of cells
should be 1 9 106, this dilution was given intraperitoneally.
Let the tumour grow in the mice for minimum 7 days before
starting treatments (Unnikrishnan et al., 1990).
LD50 determination
The toxicological profile of the most promising synthesized
antitumour derivatives (5e, 5c, and 5d) were determined by
the standard LD50 method in mice (Girgis et al., 2007).
Albino mice weighing 20–25 g were divided in 12 groups
of 6 mice each. Administration of the tested compounds (5e,
5c, and 5d) dissolved in 1 % aqueous solution of Tween 80
at doses of 500, 750 and 1,000 mg/kg (body weight) was
done intraperitoneally. The control groups were given only
vehicle. The toxic symptoms, mortality rates and the post-
mortem findings in each group were recorded 24-h post-
administration. LD50 values of the tested compounds were
calculated according to the following formula.
Treatment protocol
Swiss Albino mice were divided into eight group of six
each. All the animals in seven groups were injected with
DLA cells (1 9 106 cells per mouse) intraperitoneally and
the remaining one group is a normal control group.
Group 1
Group 2
served as the normal control.
served as the tumour control. Group 1 and 2
receives normal diet and water.
P
ðZXdÞ
Group 3
served as the positive control was treated
with injection fluorouracil at 20 mg/kg body
weight intraperitoneally.
LD50 ¼ Dm ꢁ
n
Where Dm is the largest dose which killed all animals, z is
the mean of dead animals between two successive
Group 4–9
served as a treatment control group and was
administered synthetic derivatives (5a–f) in a
dose of 50 mg/kg dissolved with 0.5 ml
DMSO administered through intraperitoneally.
Table 1 Physical characterization of all the synthesized derivatives
Compound
code
Colour
Molecular
formula
M.p.(°C) %Yield
In this study, drug treatment was given after 24 h of
inoculation once daily for 14 days. On day 14, after the last
dose, all mice from each group blood were withdrawn from
each mouse by retro orbital puncture or bleeding and the
parameters such as haematological, serum enzyme, serum
lipid, body weight, life span (%) and cancer cell count were
determined
2a
3a
4a
4b
4c
4d
4e
4f
Pale rose
C9H10N2O
179–180 78
129–131 70
244–246 83
226–227 92
230–231 90
140–142 81
261–262 79
271–272 69
255–258 77
265–267 77
223–225 67
210–213 76
239–240 66
200–203 61
Pale brown C9H8N2O
Pale yellow C16H12N2O
Yellow
C16H11ClN2O
Pale yellow C17H14N2O2
Brick red
Brown
C
C
C
C
18H17N3O
16H11N3O3
16H12N2O2
20H14N4O3
Evaluation of clinical parameters
Pale brown
Brown
Cancer cell count
5a
5b
5c
5d
5e
5f
Pale yellow C20H13ClN4O3
The fluid (0.1 ml) from the peritoneal cavity of each mouse
was withdrawn by sterile syringe and diluted with 0.8 ml of
ice cold normal saline or sterile phosphate buffer solution
and 0.1 ml of trypan blue (0.1 mg/ml) and total numbers of
the living cells were counted using haemocytometer. The
Pale green
Brown
C
C
C
21H14N4O3
20H14N4O3
20H14N4O3
Yellow
Pale yellow C20H14N4O3
123

Med Chem Res
Fig. 2 Proposed mechanism for
the formation of titled
derivatives (5a–f)
..
O
:
N
CH3COOH
NH
H
N
H
O+
..
N
O
HN
NH
CH3COO-
+
O
O
H
H
O
HN
NH
O
H
N
O
O
O
NH
O
HO
-H2O
N
HN
N
N
H
parameters such as haematological profile, serum enzyme
level, lipid profile, change in the body weight and per-
centage increase of life span were evaluated (Santhosh
et al., 2007). In Hodgkin lymphoma, high cholesterol level
and low triglyceride level have been reported. Hence,
we investigated this parameter for our current study
(Jacqueline et al., 1998).
carbanion. The electron-rich carbanion unit can make a
possible attack on the carbonyl carbon of the a, b unsatu-
rated ketone of benzimidazole unit and forms a adduct.
Finally, on dehydration of the adduct can form a carbon–
carbon double bond between the barbituric acid and a, b
unsaturated ketone of benzimidazole unit. The proposed
mechanism is shown in the Fig. 2. The structures of the all
final derivatives were established on the basis of spectral
analysis. IR spectrum of the 5-[(2E)-1-(1H-benzimidazol-2-
yl)-3-phenylprop-2-en-1-ylidene] pyrimidine-2,4,6(1H,3H,
5H)-trione 5b showed a strong absorption band at
3,232 cm-1 corresponding to benzimidazole NH,absorption
at 2,910 and 2,878 cm-1 corresponds to pyrimidine NH/NH.
A sharp absorption at 1,678 cm-1 corresponds to carbonyl
stretching and 771 cm-1 due to aryl chloride. The 1H NMR
spectra showed the singlet peak at d 8.4 and 8.6 were
assigned to pyrimidine NH/NH. The singlet peak at d 8.9
corresponding to benzimidazole NH. The doublet for vicinal
protons has seen along with the aromatic multiplet between
d 6.4 and 8.3 ppm. Mass spectrum of compound 5b revealed
the molecular ion peak [M?2] at m/z 394 corresponding to
the molecular mass of the compound (Table 2). Mass frag-
mentation pattern of 5b gave a base peak of m/z 58 which
corresponds to the molecular formula of CH2N2O. This
Result and discussion
Chemistry
A typical synthetic strategy was employed to obtain the
titled compounds (5a–f). The present study described a
knoevenagel condensation between a benzimidazole chal-
cone and barbituric acid by means of the experimental
protocol as shown in Fig. 1. The chemistry involves the
formation of final derivatives: a, b unsaturated benzimid-
azole ketone initially undergoes protonation by acetic acid.
The protonated form of the ketone facilities the addition
reaction towards a nucleophile. The carboxylate anion
which is formed in the former step can accept a proton from
the methylene unit of barbituric acid and generate a
123

Med Chem Res
Table 2 Spectral characterization of all the final barbitone derivatives (5a–f)
Compound
FT-IR (KBr, Vmax cm-1
)
1HNMR (DMSO-d6),d ppm
m/z
5a
3,230 (NHstr), 1,693 (C=O), 1,587
(C=N)
9.1 (1H,s,NHbenzimidazole), 8.0–8.4 (2H,s,pyrimidineNH),
6.4–8.1 (11H,m,9ArH,2CH=CH)
359.21 (M??1)
5b
5c
5d
5e
5f
3,232 (NHstr), 1,678 (C=O), 1,581
(C=N), 771 (Ar–Cl)
8.9 (1H,s,NHbenzimidazole), 8.4–8.6 (2H,s,pyrimidineNH),
6.4–8.3 (10H,m,8ArH,2CH=CH)
394.1 (M??2).
390.06 (M??1)
402.56 (M??1)
405.56 (M??2)
375.65 (M??1)
3,239 (NHstr), 1,681 (C=O), 1,576
(C=N)
8.9 (1H,s,NHbenzimidazole), 8.4–8.6 (2H,s,pyrimidineNH),
6.4–8.3 (10H,m,8ArH,2CH=CH), 3.3 (3H,s,OCH3)
3,227 (N–H str), 1,678 (C=O),
1,569 (C=N)
8.9 (1H,s,NHbenzimidazole), 8.5–8.7 (2H,s,pyrimidineN),
6.3–8.4 (10H,m,8ArH,2CH=CH), 3.3 (6H,s,N(CH3)2
3,227 (NHstr), 1,678 (C=O), 1,569
(C=N), 1,310 (Ar–NO2)
8.5 (1H,s,NHbenzimidazole), 8.2–8.4 (2H,s,pyrimidine NH),
6.4–8.4 (10H,m,8ArH,2CH=CH)
3,379 (ArOH), 3,231 (NHstr),
1,680 (C=O), 1,562 (C=N)
9.1 (1H,s,NHbenzimidazole), 8.4–8.8 (2H,s,pyrimidineNH),
6.4–8.5 (10H,m,8ArH,2CH=CH), 5.62 (1H,s,ArO–H)
Effect on biochemical parameters
spliting of the urea fragment from the 5b also support with
the formation of final barbitone moiety.
The inoculation of DLA cells caused significantly increase
in the level of total cholesterol, aspartate amino transferase,
alanine amino transferase, alkaline phosphatase in the
tumour control animals (G2), when compared to the normal
group. The treatment with various synthetic compounds
(5a–f) at dose of 50 mg/kg body weight reversed these
changes towards the normal level which showed in
Table 4. All the values were found to be significant. The
treatment with standard 5-FU also gave similar results.
Antitumour activity
Effect on haematological parameters
The haematological parameters such as RBC, haemoglobin
and platelets were decreased and WBC count was signifi-
cantly increased in the DLA control group when compared
to the normal control group (Table 3). Treatment with
various synthetic compounds such as 5a–f at dose of
50 mg/kg body weight significantly increases the haemo-
globin content, RBC, platelets and significantly decreased
the WBC count to about normal level. All these results
suggest the antitumour nature of the synthetic derivatives.
However, the standard 5-FU at the dose of 20 mg/kg body
weight produced a better result in all these parameters.
Effect on tumour growth
In the DLA tumour control group, the average life span of
animal was found to be 50 % whereas synthetic drug such
as 5a–e at dose of 50 mg/kg body weight increase the life
span to 66, 70, 72, 74, 76 and 69 %, respectively. These
values were significant. However, the average life span of
Table 3 Effect of final titled derivatives (5a–f) on haematological parameters
Treatment
Total WBC cells (ml 9 103)
RBC count (Mill/mm3)
Hb (Gm/dl)
PCV (%)
Platelets (Lakhs/mm3)
G1
G2
G3
G4
G5
G6
G7
G8
G9
10.45 1.30
4.30 0.90
12.40 1.12
14.65 2.32
3.15 0.72
14.40 2.50a**
11.30 1.72b**
13.80 2.22b*
12.70 1.92b**
12.55 1.82b**
12.50 1.60b**
12.45 1.48b**
13.45 1.28b**
2.46 0.45a**
4.10 0.80b**
2.90 0.45b*
3.10 0.65b**
3.15 0.60b**
3.25 0.70b**
3.30 0.80b**
2.80 0.70b**
7.12 0.95a**
11.5 1.45b**
9.30 0.99b*
10.30 1.45b**
10.28 1.45b**
10.42 1.45b**
10.50 1.60b**
10.10 1.68b**
30.55 3.52a**
17.45 1.35b**
24.10 2.65b*
23.10 1.32b**
22.90 1.35b**
22.42 1.32b**
20.48 1.22b**
23.68 1.32b**
1.60 0.52a**
2.65 0.72b**
1.99 0.42b*
2.10 0.69b**
2.12 0.55b**
2.22 0.60b**
2.30 0.66b**
2.05 0.76b**
All values are expressed as mean SEM for 6 animals in each group
G1 normal control, G2 cancer control, G3 positive control, G4 treatment control (5a), G5 treatment control (5b), G6 treatment control (5c), G7
treatment control (5d), G8 treatment control (5e), G9 treatment control (5f)
a**
Values are significantly different from control (G1) at P \ 0.01
b*
Values are significantly different from cancer control (G2) at P \ 0.05
b**
Values are significantly different from cancer control (G2) at P \ 0.01
123

Med Chem Res
ALP (U/L)
Table 4 Effect of final titled derivatives (5a–f) on serum enzymes and lipid proteins
Treatment
Cholesterol (mg/dl)
TGL (mg/dl)
AST (U/L)
ALT (U/L)
G1
G2
G3
G4
G5
G6
G7
G8
G9
98.10 3.48
121.82 2.45
38.48 1.22
32.30 1.26
124.12 2.22
140.86 4.60a**
112.52 3.88b**
136.45 3.62b*
129.32 3.55b**
126.35 2.90b**
124.30 2.50b**
122.30 2.52b**
131.30 2.57b**
210.12 4.65a**
156.42 2.78b**
186.85 2.32b*
182.50 2.40b**
170.60 2.50b**
172.40 2.38b**
168.70 2.43b**
178.70 2.83b**
85.6 2.72a**
56.30 1.60b**
77.60 2.25b*
76.52 2.60b**
75.62 2.70b**
73.40 2.52b**
72.60 2.60b**
75.40 2.68b**
61.20 2.60a**
42.45 1.60b**
57.30 1.88b*
55.60 1.72b**
53.52 1.62b**
51.45 1.45b**
53.42 1.65b**
56.42 1.34b**
242.22 4.30a**
162.30 2.33b**
232.32 2.36b*
223.36 2.60b**
218.26 2.42b**
210.21 2.64b**
202.85 2.60b**
229.15 2.68b**
All values are expressed as mean SEM for 6 animals in each group
TGL triglycerides, AST aspartate amino transferase, ALT alanine amino transferase, ALP alkaline phosphatase
a**
Values are significantly different from control (G1) at P \ 0.01
b*
Values are significantly different from cancer control (G2) at P \ 0.05
b**
Values are significantly different from cancer control (G2) at P \ 0.01
Table 5 Effect final titled derivatives (5a–f) on the life span, body weight and cancer cell count of tumour induced mice
Treatment
No. of animals
% ILS life span
Increase in body weight (g)
Cancer cell count (ml) 9106
G1
G2
G3
G4
G5
G6
G7
G8
G9
6
6
6
6
6
6
6
6
6
ꢂ30 days
50 %
2.20 0.55
–
7.60 0.94a**
3.70 0.50b**
6.45 0.78b*
6.60 0.54b**
6.30 0.44b**
5.90 0.40b**
5.88 0.38b**
5.43 0.28b**
2.76 0.33
1.30 0.32b**
2.26 0.42b*
2.12 0.45b**
1.96 0.42b**
1.90 0.32b**
1.88 0.28b**
2.03 0.48b**
94 %
66 %
70 %
72 %
74 %
76 %
69 %
All values are expressed as mean SEM for 6 animals in each group
a**
Values are significantly different from control (G1) at P \ 0.01
b*
Values are significantly different from cancer control (G2) at P \ 0.05
b**
Values are significantly different from cancer control (G2) at P \ 0.01
5-FU treatment was found to be 94 %, indicating its potent
antitumour nature. It was also supported by the significant
reduction in packed cell volume and viable. Tumour cell
count in various synthetic compounds at dose of 50 mg/kg
treatment when compared to the DLA tumour control
(Table 5).
Treatment with various synthetic compounds such as 5a–
f at dose of 50 mg/kg body weight inhibited the tumour
volume, viable tumour cell count and increased the life
span of the tumour-bearing mice. The reliable criteria for
judging the value of any anticancer drug are the prolon-
gation of the lifespan of animals. It may be concluded that
various synthetic compounds, by decreasing the nutritional
fluid volume and arresting the tumour growth, increase the
life span of DLA-bearing mice. Thus, various synthetic
compounds such as (5a–f) at dose of 50 mg/kg have anti-
tumour activity against DLA-bearing mice. Usually, in
cancer chemotherapy, the major problems which are being
encountered are of myelo suppression and anaemia. The
anaemia encountered in tumour bearing mice is mainly due
to the reduction in RBC or haemoglobin percentage and
Conclusion
In DLA tumour-bearing mice, a regular rapid increase in
Ascitic tumour volume was observed. Ascitic fluid is the
direct nutritional source for tumour cells and a rapid
increase in Ascitic fluid with tumour growth would be a
means to meet the nutritional requirement of tumour cells.
123

Med Chem Res
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this may occur either due to iron deficiency or due to
haemolytic or myelopathic conditions. Treatment with all
synthetic compounds such as (5a–f) brought back the
haemoglobin (Hb) content, RBC and WBC count more or
less to normal levels significantly. This clearly indicates
that 5-[(2E)-1-(1H-benzimidazol-2-yl)-3-substituted phe-
nylprop-2-en-1-ylidene] pyrimidine-2,4,6(1H,3H,5H)-tri-
ones possess protective action on the haemopoietic system.
It was reported that the presence of tumour in the human
body or in the experimental animals is known to affect may
function of the liver. The significantly elevated level of total
cholesterol and enzyme level in serum of tumour inoculated
animal indicated liver damage and loss of functional integ-
rity of cell membrane. The significant reversal of these
changes towards the normal by various synthetic drugs
treatments. In the present study, the biochemical examina-
tion of DLA-inoculated animals showed marked changes
indicating the toxic effect of the tumour. The normalization
of these effects observed in the serum treated with various
synthetic compounds such as (5a–f) supported the potent
antitumour and hepatoprotective effect of the derivatives.
Based on the close examination on substitution of titled
molecular templates, it may be concluded that the electron-
withdrawing group like nitro in the meta position of the
phenyl system (5e) exhibited a significant antitumour char-
acter with a percentage increased life span of 76 %. We had
also noticed that electron-donating group such as methoxyl
and dimethyl amine system in the para position of the phenyl
system favours the activity ratio. Thus, our research con-
cluded that 5-[(2E)-1-(1H-benzimidazol-2-yl)-3-substituted
phenylprop-2-en-1-ylidene] pyrimidine-2,4,6(1H,3H,5H)-
triones system can play a crucial role against the tumour
growth and further suitable derivatization of such com-
pounds on both aromatic and heteroaromatic nucleus based
up on our report hope to get more selective anticancer agents.
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Acknowledgments The authors are highly thankful to Dr.
N. Chidhambarnathan, K.M.College of Pharmacy, Madurai, India,
Mrs. Githa Elizabeth Mathew, Assistant Professor, Department of
pharmacology, Grace College of Pharmacy, Kerala, India, for carry-
ing out the pharmacological work for the present study. Authors also
acknowledge the help of Dr. N. Murugesan, SAIF IIT, Chennai, for
carrying out the spectral analysis
Sing P, Kaur M, Verma P (2009) Design, synthesis and anticancer
activity of hybrids of indole and barbituric acid-Identification of
highly promising leads. Bioorg Med Chem Lett 19:3054–3058
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