Synthesis and
Evaluation of Phenothiazine derivative for Anti-depressant activity
Nachiket S. Dighe1*,
Renuka N.Barhate1, Ravindra B. Lawre2, Sunil A. Nirmal3
1Department of
Pharmaceutical Chemistry, Pravara Rural College of Pharmacy, Loni, MS,
India-413736
2Department of Pharmaceutics, Pravara Rural College of Pharmacy, Loni, MS,
India-413736
3Department of Pharmacology, Pravara Rural College of
Pharmacy, Loni, MS, India-413736
*Corresponding Author E-mail: nachiket1111@rediffmail.com
ABSTRACT:
This
study was designed to synthesize, characterize and to evaluate the
pharmacological activity of substituted phenothiazine derivatives. Totally
thirteen compounds, were synthesized by conventional method. Purity of the
synthesized compounds was ascertained by TLC and melting point determination by
open capillary tube method and they were characterized by IR and NMR
spectroscopic methods. Antidepressant activity of all the synthesized compounds
was evaluated by despair swim test by using Sprague Dawley Rats. Standard drug
Imipramine was used as the control. In the results of the spectral study. all
the compounds showed characteristic peak in IR and NMR spectroscopy. In the
despair swim test, all the synthesized derivatives showed antidepressant
activity. Among them three Compounds (A1,
A3 A6 and A8) showed significant antidepressant
activity comparing with control drug imipramine. These results are useful for
the further investigation in the future.
KEYWORDS: Polyamide 6, Degradation
mechanism, Flame retardants, Coats-Redfern method.
Current
treatments for depression either fail to produce recovery or induce unwanted
side effects. So there is still a large unmet clinical need(1-2).
The heterocyclic compounds which contain nitrogen and sulphur possess an
enormous significance in the field of medicinal chemistry.(10) The
main aims in the development of new antidepressants were greater efficacy,
absence of side effects, lack of toxicity in over dose and earlier onset of
action(3).
Elaborate
research work has been carried out in the past and continuing in the present to
synthesize new compounds to meet this depression. The forced swim test
(behavioral despair test) and tail suspension test in the rat are widely used
for the initial screening of antidepressants. These tests have good predictive
validity and allow rapid and economical detection of substances with potential
antidepressant like activity. The tests are base on the same principle:
measurement of the duration of immobility when rodents are exposed to an
inescapable situation. The majority of clinically used antidepressants decrease
the duration of immobility(4). Depression is a serious medical issue
characterized by a variety of debilitating symptoms, such as persistent sadness
and anxiety, chronic fatigue, feelings of worthlessness, disturbances in
cognitive functioning and thoughts and attempts of suicide(5).
Depression has been determined to be the leading cause of disability and the
4th leading contributor to the global burden of disease and is characterized by
relapse, recurrence and chronicity(6). Antidepressants are the drugs
used to treat depression thereby elevates mood and modifies the behavior. Half
a century ago, antidepressants were discovered by serendipity(7).
As estimated by WHO, depression shall become the second
largest illness in terms of morbidity by another decade in the world; already
one out of every five women, and every twelve men have depression. Not only
adults, but two percent of school children, and five percent of teenagers also
suffer from depression, and these mostly go unidentified.[11]
To the present knowledge, antidepressant drugs used in
the treatment of major depressive disorders are believed to act on the central
monoaminergic systems mainly serotonin (5-hydroxytryptamine, 5-HT) and
noradrenaline (NA) synaptic neurotransmissions. Selective serotonin reuptake
inhibitors (SSRIs: paroxetine, fluoxetine, citalopram, escitalopram,
fluvoxamine, sertraline) and noradrenaline reuptake inhibitors (NRIs:
reboxetine, desipramine) are the most common prescribed antidepressant drugs.[12]
2. Materials and methods
2.1 Chemistry
Melting
points were determined in open capillary method and are uncorrected. The 1H-NMR
spectra were recorded on sophisticated multinuclear FT-NMR Spec-trometer model
Advance-II (Bruker) using dimethylsulfoxide-d6 as solvent and
tetramethylsilane as internal standard. IR spectra were recorded on Jasco
FT-IR-spectrophotometer using KBr disc method. Antidepressant activity of all
the synthesized compounds was evaluated by despair swim test using Sprague Dawley Rats.
2.2.1General procedure for the preparation of 7, 8 or
9 substituted aniline Benzoic acid derivatives (I)
Equimolar amount of
substituted aniline was added to a chloro benzoic acid in 20 mL of DMF and 0.1 percent of
potassium hydroxide solution and the
reaction mixture was heated under refluxed
at about 80°C temperature, for 2 h. TLC indicated the end of reaction.
The mixture was cooled by addition of a water/ice mixture. The solid was
filtered in excellent yield .
2.2.2General procedure for the preparation of 7, 8 or
9 substituted 10H-phenothiazine 1 carboxylic acid derivatives (II)
Equimolar amount of 7, 8 or
9 substituted Anilino Benzoic acid was added to a solution of sulfur powder and
iodine in 5 mL of ethanol. Reaction mixture was heated under reflux with
stirring for about 2 h and poured into ice/water mixture. The precipitate was
filtered and washed with cold water (A1-A13).
2.3 Scheme:
2.3 Pharmacology:
Rat-Sprague
Dawley (220-255 gm), 8-12 weeks old, was obtained from National Institute of
Bioscience, Pune. They were housed in autoclaved polypropylene cages in groups
of 2-3 rats per cage and kept in a room maintained at 19 to 25 0 C
and humidity 45 to 65 % with a 12-h light/dark cycle. They were allowed to
acclimatize for four days before the experiments and were given free access to
Standard sterilized extruded rodent diet was provided ad libitum, Reverse Osmosis water treated with UV light was
provided ad libitum in autoclaved
polypropylene bottles and Autoclaved corn cob was used as bedding material.
All
procedures of the present study was in accordance with the standard operating
procedures of the Prado Pvt. Ltd. guidelines provided by the Committee for the
Purpose of Control and Supervision of Experiments on Animals (CPCSEA) as
published in The Gazette of India, December 15, 1998. Prior approval of the
Institutional Animal Ethics Committee (IAEC) was obtained before initiation of
the study (IAEC-13-002).
2.3.1 Antidepressant activity (forced swim test in
rat):
Behavioral
despair or forced swim test (FST) was proposed as a model to test
antidepressant activity by Porsolt et al. (1977) (8-9). It was
suggested that mice or rats when forced to swim in restricted space from where
they cannot escape are induced to a characteristic behavior of immobility. This
behavior reflects a state of despair which can be reduced by several agents
which are therapeutically effective in human depression. The behavioral despair
test is employed to assess the antidepressant activity of synthesized
derivatives. Sprague-Dawley rats of 200-270 gm in a group of two each were used and on
the first day of the experiment (pretest session), rats were individually
placed in a cylindrical recipient (Plexiglass cylinder) of dimensions
(diameter, 10 cm; height, 25 cm) containing 10 cm of water 25°C. The animals
were left to swim for 6 min before being removed, dried and returned to their
cages. The procedure was repeated 24 h later, in 5 min swim session (test
session). The synthesized compounds (25 mg kg-1), and imipramine, as a
reference antidepressant drug (25 mg kg-1) were suspended in a 0.5 % aqueous
solution of Na CMC (Corboxy Methyl Cellulose). The drugs were given by gavage
in a standard volume of 10ml/kg body weight, 1 h prior to the test. Control
animals received 0.5 % aqueous solution of Na CMC (Corboxy Methyl Cellulose). This test was performed after 1 hr, 5 hrs and 24 hrs of dose
administration. For individual animal video recording was made.Then, the rats were dropped individually into the
Plexiglass cylinder and left in the water for 6 min. After the first 2 min of
the initial vigorous struggling, the animals were immobile. An immobility time
is the time spent by rat floating in water without struggling, making only
those moment necessary to keep the head above the water. The total duration of
immobility was recorded during the last 5 min of the 6 min test session.
3. RESULT AND
DISCUSSION:
3.1 Chemistry
The
structures, yields and melting points of the compounds have been given in the
(Table 1).Melting points of the synthesized compounds were sharp indicating
that the compounds were pure; the yield value of the compounds also suggested
that the chemical methods were reliable for the synthesis of the compound. All
spectral data were in accordance with assumed structures.
Table
no. 01: Analytical and physicochemical data of the synthesized compounds (A1-A13)
|
Comp. |
Mol. Formula |
Mol. Wt. |
M.P. ° C |
Yield % |
Elemental analyses Calculated
|
||
|
C |
H |
N |
|||||
|
A1 |
C13H8N2O4S |
288.29 |
184 |
65 |
54.16 |
2.80 |
9.72 |
|
A2 |
C13H8N2O4S |
288.29 |
180 |
62 |
54.16 |
2.80 |
9.72 |
|
A3 |
C13H8N2O4S |
288.29 |
180 |
65 |
54.16 |
2.80 |
9.72 |
|
A4 |
C13H8ClNO2S |
277.72 |
174 |
52 |
56.22 |
2.90 |
5.04 |
|
A5 |
C13H7N3O6S |
333.72 |
194 |
60 |
46.85 |
2.12 |
12.61 |
|
A6 |
C13H8ClNO2S |
277.72 |
170 |
52 |
56.22 |
2.90 |
5.04 |
|
A7 |
C13H7Cl2NO2S |
312.17 |
172 |
70 |
50.02 |
2.26 |
4.49 |
|
A8 |
C13H7CIN2O4S |
322.72 |
193 |
60 |
48.38 |
2.19 |
8.68 |
|
A9 |
C13H7Cl2NO2S |
312.17 |
170 |
65 |
50.02 |
2.26 |
4.49 |
|
A10 |
C13H6BrN3O6S |
412.17 |
192 |
62 |
37.88 |
1.47 |
16.19 |
|
A11 |
C13H8BrNO2S |
322.17 |
180 |
65 |
48.46 |
2.50 |
4.35 |
|
A12 |
C13H7Cl2NO2S |
312.17 |
170 |
52 |
50.02 |
2.26 |
4.49 |
|
A13 |
C13H8ClNO2S |
277.72 |
175 |
60 |
56.22 |
2.90 |
5.04 |
Spectral
Data:
A1: IR (KBr) cm-1: 3010.23 (Ar-CH str.),1715.11
(-C=O str.), ,3300.23(OH str),3100.11 (CH str),1360.35) (nitro symetry), . 1H
A2: IR (KBr) cm-1: 3050.23 (Ar-CH str.),1750.12
(-C=O str.), ,2510.23(OH str),3020.11 (CH str),1295.30 (nitro symetry), 1H NMR: 1.74(carboxylic
acid),7.16(1H-phenothiazine),7.58-8.01(benzene) ,0.25 1-N=O=O from benzene
A3: IR (KBr) cm-11700.11 (-C=O str.), 1640.32 (-C=C str), 1H NMR1.74 (carboxylic
acid),7.16(1H-phenothiazine),7.58-8.01(benzene) ,0.25 1-N=O=O from benzene
A4: IR (KBr) cm-13010.23 (Ar-CH str.), 1710.11 (-C=O str.),
1550.32 (-C=N str), 850.22 (-C-Cl str),:
1H NMR:
1.74(carboxylic acid),7.16(1H-phenothiazine),7.58-8.01(benzene),0.52 1-cl from
1 benzene
A5: IR (KBr) cm-1: 3010.23 (Ar-CH str.), 1682.11 (-C=O
str.), 1355.40(nitro sym) 1H NMR: 1.74(carboxylic
acid),7.16(1H-phenothiazine),7.58-8.01(benzene),0.39 (2H-N=O=O from benzene)
A6: IR (KBr) cm-1: 3012.25 (Ar-CH str.), 1702.21
(-C=O str)850.22(C-Clstr) 1H NMR: 1.74(carboxylic
acid),7.16(1H-phenothiazine),7.58-8.01(benzene) ,0.52 1-cl from 1 benzene
A7: IR (KBr) cm-1:
945.20 (-C-Cl str), 3110.23
(Ar-CH str.), 1682.11 (-C=O
str.), 930.50 (C-Cl str) 1H NMR1.74
(carboxylic acid), 7.16(1H-phenothiazine), 7.58-8.01(benzene)0.01-0.06(2H 1 cl
from benzene)
A8: IR (KBr) cm-1: 3010.23 (Ar-CH str.), 1682.11 (-C=O
str.) 1550(nitro) 3616.11 (-OH str.). 1H NMR: 1.74(carboxylic
acid),7.16(1H-phenothiazine),7.58-8.01(benzene)0.01-0.21 (2H- Cl from
benzene),0.93 1-c(=o)o from benzene
A9: IR
(KBr) cm-1: 3010.23 (Ar-CH str.),
1682.11 (-C=O str.), 3616.11 (-OH str.), 855.56(C-Cl str).1H NMR:
1.74(carboxylic acid), 7.16(1H-phenothiazine), 7.58-8.01(benzene) 0.01-0.06(2H
1 cl from benzene
A10: IR (KBr) cm-1: 3010.23 (Ar-CH str.), 1689.78 (-C=O
str), 1460(nitro sym) 940.21. 1H NMR: 1.74(carboxylic acid),
7.16(1H-phenothiazine),7.58-8.01(benzene)
A11: IR (KBr) cm-1:3010.23 (Ar-CH str.), 1689.78 (-C=O
str), 3600.12 (-OH str.), 940.21(C-Cl
str) 1H NMR: 1.74(carboxylic acid),7.16(1H-phenothiazine), 7.58-8.01(benzene),0.17-
1 Br from benzene
A12: IR (KBr) cm-1:3210.23
(-CH=CH str.), 3020.23 (Ar-CH str.), 1750.78
(-C=O str),935.11 (C-Cl str). 1H NMR: 1.74(carboxylic
acid), 7.16(1H-phenothiazine), 7.58-8.01(benzene) 0.01-0.06(2H 1 cl from
benzene
A13: IR (KBr) cm-1: 3110.23 (Ar-CH str.), 1689.78 (-C=O str), 934.12(C-Cl str). 1H NMR: 1.74(carboxylic acid), 7.16(1H-phenothiazine),
7.58-8.01(benzene) 0.52 1-cl from 1 benzene.
3.2 Antidepressant Activity
All
the synthesized compounds were subjected to antidepressant activity study on Sprague-Dawley rats by despair
swim test. Imipramine was used as standard control. The animals show more
stable levels of immobility during the last four minutes of the session. The
results showed that all the compounds showed antidepressant activity. Among
them three Compounds (A1, A3, A6 and A8)
showed significant antidepressant activity comparing with standard control
imipramine (Table 02).
Table
03: Antidepressant activities of the compounds
|
Compound code. |
Immobility time |
% Immobility |
||||
|
1 Hr |
5 Hr |
24 Hr |
1 Hr |
5 Hr |
24 Hr |
|
|
A1 |
162.5 |
162.5 |
172 |
97.15 |
89.56 |
88.82 |
|
A2 |
156.5 |
160 |
167.5 |
92.56 |
87.21 |
85.41 |
|
A3 |
176 |
175.5 |
184.5 |
102.84 |
96.58 |
95.01 |
|
A4 |
142.5 |
156 |
163.5 |
85.84 |
85.47 |
83.84 |
|
A5 |
144 |
158.5 |
163.5 |
87.72 |
86.84 |
83.84 |
|
A6 |
164 |
164 |
173 |
96.34 |
89.86 |
88.71 |
|
A7 |
145.5 |
158 |
167.5 |
83.59 |
86.57 |
85.89 |
|
A8 |
160.5 |
163.5 |
173 |
95.34 |
89.90 |
88.74 |
|
A9 |
169.5 |
178.5 |
184 |
97.54 |
97.80 |
94.35 |
|
A10 |
151.5 |
156 |
165 |
80.08 |
85.47 |
84.61 |
|
A11 |
169 |
170.5 |
178 |
99.68 |
97.09 |
91.88 |
|
A12 |
165 |
168.5 |
176 |
97.51 |
94.78 |
90.23 |
|
A13 |
170 |
179.5 |
182 |
98.25 |
95.06 |
92.82 |
|
Control |
172 |
182.5 |
195 |
100 |
100 |
100 |
|
Imipramine (std.) |
136.5 |
150.5 |
154.5 |
79.41 |
82.49 |
79.26 |
4. DISCUSSION:
Statistical
evaluation of the equations is in accepted range. The correlation coefficient
is high with less standard error. The residual value and residual variance for
each series also is less indicating good predictive power of models. From
equation it is observed that two
electronic parameters Dipole Moment Z Component (Whole Molecule) and VAMP HOMO
(Whole Molecule) as well as one steric parameter Inertia Moment 2 Length (Whole
Molecule) contribute (-0.227, –1.469 and – 0.414 respectively) negatively for
the activity so electron withdrawing and less bulky groups may enhance the
activity (% Immobility).
5. CONCLUSION:
We
investigated the importance of functional group substitutions, in the
structural framework of the compounds for their antidepressant activity. All
compounds showed significant antidepressant activity at dose (25 mg/kg). The
compounds A4, A5 and A7) showed better
activity. Finally, the encouraging result of the antidepressant activity
displayed by these compounds may be of interest for further structural
modifications to the lead compound and next level studies in the hope of
finding a new potent antidepressant prescription.
6. REFERENCES:
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Received
on 12.12.2015 Modified on 22.12.2015
Accepted
on 29.12.2015 © AJRC All right
reserved
Asian J. Research Chem. 8(12): December 2015; Page 745-750
DOI: 10.5958/0974-4150.2015.00120.0