Analgesic,
antipyretic, CNS and pass prediction
Activities of 4-(4-hydroxy benzylidene
amino) phenol Schiff base
Valli.G.1*, MareeswariP.1, RamuK.1
and Thanga Thirupathi A.2
1Department
of Chemistry, S.F.R College for Women, Sivakasi
2Department
of Pharmacology, SB College of Pharmacy,
Anaikuttam, Sivakasi.
ABSTRACT:
The Schiff base 4-(4-hydroxy benzylidene
amino)phenol was prepared from
p-hydroxybenzaldehyde and 4-aminophenol by condensation method using
standard procedure. Analgesic,
antipyretic and CNS activities of 4-(4-hydroxy benzylidene
amino) phenol Schiff base were studied
using albino rats of both the sexes. Animals were divided into three
groups, each consisting of four animals. Group 1 served as control and group 2
received standard drug. Group 3 received 250 mg/kg of 4-(4-hydroxy benzylidene amino) phenol.
For the determination of antipyretic activity, pyrexia was induced by
20% yeast suspension. The analgesic activity was determined by tail immersion
method. CNS depressant activity of the compound was measured by placing the rat
individually in the actophotometer for 10 min. The
results obtained showed that the Schiff base was found to exhibit analgesic,
antipyretic and CNS activities. The
analgesic activity of the Schiff base compared to the standard drug pentazocine was found to be higher in the first hour and then
the activity decreases slowly. The reduction in the rectal temperature for this
compound was observed to be less than the standard drug paracetamol.
4-(4-hydroxy benzylidene amino)phenol Schiff base of
250mg/kg possessed CNS stimulant
activity with a probability <0.001. 4-(4-hydroxy benzylidene
amino)phenol Schiff base was found to
possess highest glutathione thiolesterase
inhibitor activity as 86% and other activities greater than 80% as per the prediction using PASS (online
bioactivity prediction software) .
Key words p-hydroxybenzaldehyde, 4-aminophenol, analgesic, antipyretic
, CNS and PASS.
INTRODUCTION:
Schiff bases are the
important compound owing to their wide range of biological activities and
industrial application. They have been found to possess the pharmacological
activities such as antimalarial[1], anticancer[2],
antibacterial[3], antifungal[4], antitubercular[5], antiinflammatary, antimicrobial[6] and antiviral[7] etc.
They also serve as a back bone for the synthesis of various heterocyclic
compounds. In view of these above biological importance of Schiff bases, we
plan to synthesis 4-(4-hydroxy benzylidene
amino)phenol Schiff base by standard procedure. The present work focus on the
synthesis of the Schiff base from p-hydroxybenzaldehyde
by condensing with 4-aminophenol in ethanol at 50-600C for two
hours. The synthesized compound have been characterized on the basis of their
melting point and IR spectral data. The
pharmacological activities like analgesic, antipyretic and CNS were studied. In
addition to these activities we plan to predict the bioactivity by using online
bioactivity predicting software PASS
PASS is a software product designed as a tool for evaluating
the general biological potential of an organic drug-like molecule. PASS provides simultaneous prediction of many
types of biological activity based on the structure of organic compounds.
MATERIALS AND METHODS:
Materials used:
The chemicals such as p-hydroxybenzaldehyde,
4-aminophenol of E. Merck grade and distilled ethanol were used. The melting
point was determined using melting point apparatus and IR spectra were recorded
in FT IR Affinity-1 Shimadzu, Japan.
Drugs:
Chlorpromazine (standard for CNS), Pentazocine
(standard for analgesic) and paracetamol (standard for antipyretic) were chosen for our
work.
Animals used:
For the analgesics, antipyretic and CNS depressant activity studies
twenty eight albino rats of both sexes of weight 100-165g were used for each
studies.
Software used:
Chem ultra 11.0 and PASS.
Methods used:
Preparation of Schiff bases:
The p-hydroxybenzaldehyde and 4-aminophenol
were taken in a equimolar ratio of 0.01mol and
refluxed with ethanol for 2hours. After refluxing, the product obtained was
filtered, dried and recrystallized using ethanol. The
synthesized compound was used to study the
analgesic, antipyretic and CNS activities. The structure of the compound
was proved by melting point determination and IR spectral studies.
The melting point was recorded using melting point apparatus. The
melting point of 4-(4-hydroxy benzylidene
amino)phenol Schiff base observed as 285oC and the IR spectral
studies reveals the presence of C-N, C=N & C-O functionalities and showed
the following stretching frequencies. C-N Stretching at u1250cm-1, C=O stretching at u1670cm-1, C=N stretching at u1570cm-1 and C-O stretching was u1300 cm-1.
DETERMINATION OF ANALGESIC ACTIVITY [8]
The tail immersion test was carried out as described by standard
procedure. The albino rats were selected and last
3.5 cm of their tail was immersed in hot water thermo-statistically maintained
at 55° C, a procedure that caused them to rapidly withdraw their tail. Three
groups of animals were held in position in a suitable restrainer with the tail
extending out. The latency to withdraw the tail was recorded with a stopwatch,
and a cut-off maximum latency of 15 sec was established in order to prevent
tissue damage. Group I served as control, which received only vehicle (5 mg/kg,
i.p). Other groups of animals received one of the
following in a similar manner: Pentazocine (4 mg/kg,i.p) and 4-(4-hydroxy benzylidene
amino)phenol Schiff base (250mg/kg, p.o). The initial
reading was taken immediately before administration of test samples and then at
1, 2, 3 and 4 hours after the administration and the recorded data were listed
in Table-1.
ANTIPYRETIC ACTIVITY DETERMINATION [9, 10]:
Three groups of four animals of
albino rats of both sexes of weight 100-165g were used for the study. The animals were kept in polypropylene cages
in a room maintained under controlled atmospheric conditions. The animals were
fed with standard diet (Hindustan liver, Mumbai, India) and had free access to
clean drinking water. Antipyretic activity was measured by Brewer’s induced
pyrexia model in rats. Rats were fasted overnight with water ad lib before the
experiments. Pyrexia was induced by subcutaneously injecting 20% w/v brewer's
yeast suspension (10 ml/kg) into the animals' dorsum region. Eighteen hours
after the injection, the rectal temperature of each rat was measured using a
digital thermometer .Only rats that showed an increase in temperature of at
least 0.7°C were used for the experiments. Animals were divided in to 3 groups,
each containing four animals. Group I served as control (received distilled
water), Group II received the standard drug (received paracetamol
33mg/kg, p.o) .Group III received 4-(4-hydroxy benzylidene amino)phenol schiff
base (250mg/kg, p.o). The temperature was measured at
1, 2, 3 and 4hr after drug administration. The recorded values were listed in Table-2.
CNS DEPRESSANT ACTIVITY-DETERMINATION [8]:
CNS depressant activity-determination:
Locomotor methods were employed
to determine the CNS depressant activity.
Preparation of the drug for the experimental study:
Schiff base and the standard
drugs were administered in the form of suspension in water with 1% Sodium Carboxy Methyl Cellulose (SCMC) as suspending agent.
Locomotor activity:
Locomotor activity was recorded
with using Actophotometer (digital activity cage).
The animals were divided into three groups (n = 4). Each rat was individually
placed in the actophotometer for 10 min. Animals of
group 1 were intraperitoneally treated with Caffeine
(30 mg/kg) (i.p). Group 2 was treated orally with Chlorpramazine (3 mg/kg, i.p.).
Group 3 was treated orally with 250 mg/kg dose levels of drugs. Basal reaction
time was noted before and 30 min after the administration of treatment. Account
is recorded when the beam of light falling on the photocell of actophotometer is cut off
by rat Sample 1 received reference standard Chlorpramazine
at a dose of 3 mg/kg (i.p.) 30 min before the test.
Mean change in the locomotor activity was recorded
for each group and were listed in Table- 3.
Chemdraw
ultra11.0 software:
The structure of 4-(4-hydroxy benzylidene
amino)phenol schiff base was drawn in chemultra11.0
appear as given in Fig.1. and their structure was saved as molfiles (*.mol).
Fig 1: structure
of 4-(4-hydroxy benzylidene amino)phenol Schiff base
Docking:
Their possible bioactivities with PASS software (V. Poroikov et al,
version 1.917) was predicted as given in Fig.2 and the result was given
as Table-4
Fig.2. PASS Prediction
window
RESULT AND DISCUSSION:
Analgesic activity:
The increase in the basal reaction time from 2.25 to 8
seconds for 4 mg/kg of pentazocine and 2.5 to 6.25
seconds for 4-(4-hydroxy benzylidene amino)phenol Schiff
base were observed. The 4-(4-hydroxy benzylidene
amino)phenol Schiff base was found to possessed higher activity at the first hour and lesser activity at the second
hour than the standard with a
probability <0.01.
Table -1-Effect of
4-(4-hydroxy benzylidene amino)phenol Schiff base on
reaction time (in sec) in albino rats
|
Drug
treatment |
Dose (mg/ kg) |
Basal reaction time after drug administration (in sec) |
|||
|
1hr |
2hr |
3hr |
4hr |
||
|
Control |
5mg/ kg |
1.25± 0.2886 |
1.25± 0.2886 |
1.25± 0.2886 |
1.5± 0.3331 |
|
Standard Pentazocine |
4mg/ kg |
2.25± 0.5527 (44.44%) |
6.75± 0.7264 (81.48%) |
7± 1.9436 (82.14%) |
8± 0.4711 (81.25%) |
|
4-(4-hydroxy
benzylidene amino)phenol |
250mg/kg |
2.5± 0.3331 (50.00%) |
3.5± 0.3331 (64.28%) |
5.5± 0.3333 (77.27%) |
6.25± 0.5527 (76.00%) |
One way ANOVA
|
F |
3.5 |
46.03846 |
8.95804 |
70.8260 |
|
df |
(2,9) |
(2,9) |
(2,9) |
(2,9) |
|
P |
- |
<0.01 |
<0.5 |
<0.05 |
Data are expressed as Mean± SEM, n=3 in each group, statistical
analysis done by one way ANOVA followed by Dunnett’s
test. P<0.01. The value in the parenthesis indicates the percentage of
analgesic activity.
|
Parameter |
4-(4-hydroxy benzylidene amino)phenol schiff base |
|||
|
1 hour |
2 hour |
3 hour |
4 hour |
|
|
t values |
0.4330 |
4.6843 |
0.9217 |
2.6819 |
|
P-values |
- |
<0.01 |
<0.5 |
<0.05 |
Analgesic
activity by tail immersion method
Percentage
of activity
Drugs:
Std- standard pentazocine drug, PA 250mg/kg
of 4-(4-hydroxy benzylidene amino) phenol Schiff base.
Antipyretic activity:
The reduction in
temperature from 37.75 to 36.5 for 33mg/kg for Paracetamol and 37.27 to 36.57
for 250mg/kg of 4-(4-hydroxy benzylidene amino)phenol Schiff base were observed. The
reduction in temperature of 4-(4-hydroxy benzylidene
amino)phenol Schiff base is (0.7%) was found to possessed slightly lower
antipyretic activity than that of the standard (1.2%).
Table -2
Effect of 4-(4-hydroxy benzylidene
amino)phenol Schiff base on rectal temperature(0C) in albino rats
|
Drug
treatment |
Dose
(mg/kg) |
Rectal
temperature after yeast administration (oC) |
Rectal
temperature after administration of drug (oC) |
Reduction
in temperature (oC) |
||||
|
Normal |
18 hr |
1 hr |
2 hr |
3 hr |
4 hr |
|||
|
Control
saline |
1ml/kg |
36.6± 0.2828 |
37.575± 0.2995 |
37.575± 0.2995 |
37.575± 0.2995 |
37.475± 0.2995 |
37.375± 0.2995 |
- |
|
Standard paracetamol |
33mg/kg |
36.5± 0.2000 |
37.75± 0.2185 |
37.45± 0.1971 |
37.05± 0.1971 |
36.75± 0.2380 |
36.5± 0.2000 |
1.2 |
|
4-(4-hydroxy
benzylidene amino)phenol |
250mg/kg |
36.25± 0.1452 |
37.275± 0.0550 |
37.125± 0.0550 |
36.9± 0.0816 |
36.75± 0.1104 |
36.575± 0.1363 |
0.7 |
One way ANOVA
|
F |
1.6392 |
3.7145 |
4.4185 |
6.3426 |
|
df |
(2,9) |
(2,9) |
(2,9) |
(2,9) |
|
P |
<0.5 |
- |
- |
- |
Data are expressed as Mean±SEM, n=3 in each group, statistical analysis done by
paired t test. P<0.5, compared to Paracetamol.
|
Parameter |
4-(4-hydroxy benzylidene amino)phenol schiff base |
|||
|
1 hour |
2 hour |
3 hour |
4 hour |
|
|
t values |
1.1144 |
0.4996 |
0 |
0.2226 |
|
P-values |
<0.5 |
- |
- |
- |
CNS depressant activity:
The dose dependent
depression in the locomotor activity was measured for
caffeine, chlorpromazine and 4-(4-hydroxy benzylidene
amino)phenol Schiff base. Higher stimulant activity was observed for 250mg/kg
of 4-(4-hydroxy benzylidene amino)phenol Schiff base
than that of caffeine and lower depressant activity than that of chlorpromazine
with a probability <0.001.
Table - 3
Effect of 4-(4-hydroxy benzylidene
amino)phenol Schiff base on locomotor activity (in
min) in albino rats
|
Drug
treatment |
Dose
(mg/kg) |
Before
treatment |
After
treatment |
% change
in activity |
|
caffeine |
3 mg/kg (i.p) |
109± 4.3462 |
129.75±4.5796 |
19.05 |
|
chlorpromazine |
30 mg/kg (p.o.) |
79.25± 1.9650 |
14.25± 1.5899 |
82.05 |
|
4-(4-hydroxy
benzylidene amino)phenol |
250 mg/kg (p.o) |
79.75± 2.2299 |
61± 7.4089 |
23.71 |
One way ANOVA
|
F |
41.8647 |
172.238 |
|
df |
(2,9) |
(2,9) |
|
P |
<0.001 |
<0.001 |
Data are expressed as Mean±SEM, n=3 in each
group, statistical analysis done by paired t test. P<0.05, compared to
caffeine standard.
|
Parameter |
4-(4-hydroxy benzylidene amino)phenol schiff base |
|
|
Before |
After |
|
|
t values |
6.8040 |
9.5103 |
|
P-values |
<0.001 |
<0.001 |
Table-4
PASS Prediction
of 4-(4-hydroxy benzylidene amino)phenol Schiff base.
|
S.No |
Pa |
Pi |
Activity |
|
1 |
0.860 |
0.007 |
Glutathione thiolesterase inhibitor |
|
2 |
0.847 |
0.030 |
Antiseborrheic |
|
3 |
0.838 |
0.011 |
Laccase inhibitor |
|
4 |
0.827 |
0.026 |
Arylacetonitrilase inhibitor |
|
5 |
0.824 |
0.013 |
Glucan
endo-1,6-beta-glucosidase inhibitor |
|
6 |
0.824 |
0.026 |
Glucose oxidase inhibitor |
|
7 |
0.822 |
0.007 |
Corticosteroid side-chain-isomerase
inhibitor |
|
8 |
0.818 |
0.012 |
Alkane
1-monooxygenase inhibitor |
|
9 |
0.818 |
0.023 |
Taurine dehydrogenase inhibitor |
|
10 |
0.796 |
0.012 |
Apoptosis agonist |
The PASS prediction of 4-(4-hydroxy benzylidene
amino)phenol Schiff base showed the glutathione thiolesterase
inhibitor activity as (Pa=0.860), antiseborrheic
activity as (Pa=0.847), laccase inhibitor activity
was found to possessed (Pa=0.838). Pa=0.827 for arylacetonitrilase
inhibitor activity, glucanendo-1,6-beta-glucosidase inhibitor and glucose oxidase inhibitor activities were found to have same Pa
values as 0.824. corticosteroid side-chain-isomerase
inhibitor activity shows (Pa=0.822), Pa=0.818 for both the alkane
1-monooxygenase and taurine dehydrogenase
inhibitor activities and apoptosis agonist activity was observed for this
Schiff base at (Pa=0.796).
CONCLUSION:
The Schiff base 4-(4-hydroxy benzylidene
amino)phenol was prepared from p-hydroxybenzaldehyde
and 4-aminophenol by condensation method using standard procedure. The
analgesic activity of the Schiff base compared to the standard drug pentazocine was found to be higher in the first hour and
then the activity decreases slowly. The reduction in the rectal temperature for
this compound was observed to be less than the standard drug paracetamol. 4-(4-hydroxy benzylidene
amino)phenol Schiff base of 250mg/kg possessed
CNS stimulant activity with a probability <0.001. The PASS prediction
of bioactivity have shown that 4-(4-hydroxy benzylidene
amino)phenol Schiff base was found to act as glutathione thiolesterase
inhibitor, antiseborrheic, laccase
inhibitor, arylacetonitrilase inhibitor, glucan endo-1,6-beta-glucosidase inhibitor, glucose oxidase inhibitor, corticosteroid side-chain-isomerase inhibitor, alkane
1-monooxygenase inhibitor, taurine dehydrogenase inhibitor >80% of activities and apoptosis
agonist activity was observed > 70% for this Schiff base.
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Received on 18.09.2012
Modified on 05.10.2012
Accepted on 09.10.2012
© A&V Publication all right reserved
Research J.
Science and Tech. 4(5): September –October,
2012: 192-196