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RAJIV GANDHI UNIVERSITY OF HEALTH
SCIENCES KARNATAKA, BANGALORE
M. PHARM SYNOPSIS
YEAR OF ADMISSION-JULY 2011
TITLE OF THE SYNOPSIS
“AN INVESTIGATION OF ANTI-EPILEPTIC AND ANTI-PSYCHOTIC ACTIVITIES
OF IPOMOEA RENIFORMIS IN EXPERIMENTAL ANIMALS"
BY
Mrs. CHITRA KK
M. PHARM., PART-I
DEPARTMENT OF PHARMACOLOGY
UNDER THE GUIDANCE OF
Mrs. BABITHA S M.Pharm., (Ph.D)
Lecturer
DEPARTMENT OF PHARMACOLOGY
INSTITUTION
SREE SIDDAGANGA COLLEGE OF PHARMACY
B. H. ROAD, TUMKUR-572 102.
KARNATAKA
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
KARNATAKA, BANGALORE
ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1.
NAME OF THE
CANDIDATE
AND ADDRESS
Mrs. CHITRA KK
M.Pharma Part-I
Department of Pharmacology,
Sree Siddaaganga College of Pharmacy,
B.H.Road, Tumkur – 572 102
2.
NAME OF THE GUIDE
3.
NAME OF THE
INSTITUTION
SREE SIDDAGANGA COLLEGE OF PHARMACY
B. H. ROAD, TUMKUR- 572 102
4.
COURSE OF STUDY AND
SUBJECT
MASTER OF PHARMACY (PHARMACOLOGY)
5.
DATE OF ADMISSION OF
COURSE
29th JULY 2011
6.
TITLE OF THE TOPIC
Mrs. BABITHA S M.Pharn.,(Ph.D)
Lecturer
DEPARTMENT OF PHARMACOLOGY
“AN INVESTIGATION OF ANTI-EPILEPTIC AND ANTI-PSYCHOTIC ACTIVITIES OF
IPOMOEA RENIFORMIS IN EXPERIMENTAL ANIMALS”
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BRIEF RESUME OF THE INTENDED WORK
6.1 NEED FOR THE STUDY
The term Epilepsy, based on the Greek word “epilambaein” meaning “to seize” has been
first mentioned by Hippocrates.1 Recognised from the drawn of history as ‘disease of
lightening’, it was correctly described by JH Jackson over a century ago. 2 Epilepsy is
one of the most common chronic neurological disorders with no age, racial, social,
sexual or geographical boundaries and affects about 50 million people worldwide. 3 The
prevalence is high in tropical countries, particularly in Africa, where it varies between
10 and 55 per 1000, with an estimated mean prevalence of 15. 4
In india, it affects about 7 million people. Approximately 40 % of them are women. The
prevalence of epilepsy is 0.7 % in India, which is comparable to the U.S. and other
developing nations. The estimated incidence rate range from 40 to 60 per 1000,000
population/year. The WHO estimated that approximately 80 % people with epilepsy live
in developing countries and most of them do not get adequate medical treatment. 5
Modern drug therapy of epilepsy is complicated by the inability of drugs to control
seizures in some patients and side effects that range in severity from minimal
impairment of central nervous system (CNS) to death from a plastic anemia or hepatic
failure.
6
A psychosis is the major CNS disorder which causes the serious distortion of
capacity to recognize reality, affecting approximately 10 % of worlds population. All
clinically effective antipsychotics (expect clozapine like) have potent post synaptic
dopaminergic D2 receptor blocking action. Blockade of dopamine action in corpus
striatum is responsible for the extrapyramidal symptoms (EPS) so often associated with
antipsychotic drugs . Some of the antipsychotic drugs such as chlorpromazine have a
number of adverse drug reactions such as parkinsonism (tremor, muscular rigidity and
akinesia) and many of the endocrinal and metabolic disturbances such as gynaecomastia,
galactorrhea and aggravation of diabetes. 7
Ipomoea reniformis has been used to treat diverse clinical conditions ranging from
anemia to neuronal disorders. Hence, the present project has been undertaken for
screening of the herbal drug Ipomoea reniformis,8 for anti-epileptic and anti-psychotic
3
activity in experimental animals, as Ipomoea reniformis has not been studied for its antiepileptic and anti-psychotic activity despite of its use in treatment of nervous disorders
in traditional medicines.
6.2 REVIEW OF LITERATURE
Ipomoea reniformis (Family: Convolvulaceae) is a procumbent herb spreading up to 1m
and has yellow flowers. The leaves are simple, thin, and reniform with entire margin
having auriculate base and retuse apex.
Kingdom
: Plantae
Division
: Magnoliophyta
Class
: Magnoliopsida
Order
: Solanales
Family
: Convolvulaceae
Genus
: Ipomoea
Species
: reniformis
Synonym
: Merremia emarginata 8
Synonyms
Sanskrit : Mooshakarni
Hindi
: Musakani, Chuhakani
English
: Mouse Ear Wort
Tamil
: Perettaikkirai
Telugu
: Elike jimudu, Toinnuatali
Kannada : Tigade, Valliharuhi
Distribution:
It is widely distributed all over the India, specially in damp places in upper gangetic
plain, Gujarat, Bihar, West Bengal, Western-Ghats, ascending up to 900 m in the hills,
Goa, Karnataka in India, Ceylon and Tropical Africa. It mainly grows in rainy and
winter season.9
4
Chemical Constituents
The presence of caffeic, p-coumaric, ferulic and sinapic acid esters has been identified in
seeds. Petroleum ether extract contains fats and fixed oil while aqueous extract is
reported to amino acids, tannins(condensed tannins, pseudo tannins) and starch.10
Preliminary phytochemical screening of methanolic extract of ipomoea reniformis
revealed the presence of carbohydrates, glycosides, saponins, tannins, phenolic
compounds, flavonoids.
Medicinal uses
Ipomoea reniformis has been claimed to be useful for cough, headache, neuralgia,
rheumatism, diuretic, inflammation, troubles of nose, fever due to enlargement of liver
and also in kidney diseases. Powder of leaves is used as a snuff during epileptic seizures.
Juice acts as purgative and the root is having diuretic, laxative, and applied in the disease
of the eyes and gums.11
Reported activities
The methanolic extract of this plant has proven to have anti-inflammatory12,
Antidiabetic13, antioxidant and antiobesity activities while ethanolic extract of this plant
has proven to have nephroprotective activity.
6.3 OBJECTIVES OF THE STUDY
1. Collection of Ipomoea reniformis from market.
2. To investigate the anti-epileptic activity of Ipomoea reniformis by maximal electro
shock induced seizures, isoniazid induced seizures and pentylenetetrazol induced
seizures.
3. To investigate the anti-psychotic activity of Ipomoea reniformis by apomorphine
induced stereotype behavior, pilocarpine induced purposeless chewing and
apomorphine induced climbing behavior.
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7
MATERIALS AND METHODS
7.1 Experimental animals
Adult albino wistar rats of either sex weighing approximately 180-200 g will be used.
They will be housed in standard laboratory conditions and fed with standard animal feed
and will be given water ad libitum.
7.2 Dose of Ipomoea reniformis
Suitable doses of Ipomoea reniformis will be detected as per the previously published
data
7.3 Grouping of animals
The albino wistar rat will be randomly distributed into four groups containing six
animals in each group.
Group I – control
Group II – suitable dose of standard
Group III – Ipomoea reniformis (Lower)
Group IV – Ipomoea reniformis (Higher)
7.4 Experimental procedure
Anti-epileptic study using maximal electro shock induced seizures
The albino wistar rats will be randomly distributed into four groups containing six
animals in each group. The test will be started 60 minutes after oral treatment with
suitable doses of Ipomoea reniformis or the standard. An electro convulsometer with
corneal or ear electrodes will be used to deliver the stimuli. The incidence and duration
of extensor tonus will be noted. A complete abolition of hind limb tonic extension will
be considered as 100 % protection. 14
6
Anti-epileptic study using isoniazid induced seizures
The albino wistar rats will be randomly distributed into four groups containing six
animals in each group. The groups will be treated with suitable doses of Ipomoea
reniformis or the standard. 60 minutes after the treatment with Ipomoea reniformis,
animals will be injected with subcutaneous doses of isoniazid. During the next 120
minutes the occurrence of clonic seizures and tonic seizures will be recorded. The
percentage of seizures or deaths occurring in the control group is taken as 100 %. The
suppression of these effects in the treated group is calculated as percentage of controls. 15
Anti-epileptic study using pentylenetetrazol (PTZ) induced seizures
The albino wistar rats will be randomly distributed into four groups containing six
animals in each group. The groups will be treated with suitable doses of Ipomoea
reniformis or the standard. 60 minutes after the treatment with Ipomoea reniformis,
animals will be injected with subcutaneous doses of PTZ. During the next 120 minutes
the occurrence of clonic seizures, tonic seizures and death is required. The percentage of
seizures or deaths occurring in the control group is taken as 100 %. The suppression of
these effects in the treated group is calculated as percentage of controls. 16
Anti-psychotic study using apomorphine induced stereotypy
The albino wistar rat will be randomly distributed into four groups containing six
animals in each group. The suitable dose of Ipomoea reniformis or the standard will be
administered 60 minutes prior apomorphine dosage. Apomorphine HCl (1.5 mg/kg) will
be injected subcutaneous. The animals will be placed in individual plastic cages. A 10
second observation period is used to measure the presence of stereotypic activity such
sniffing, licking and chewing 10 minutes after apomorphine administration. An animal is
considered protected if this behavior is reduced or abolished. 17
7
Anti-psychotic study using pilocarpine induced purposeless chewing
The albino wistar rat will be randomly distributed into four groups containing six
animals in each group. The groups will be treated with suitable doses of Ipomoea
reniformis or the standard. 60 minutes after the treatment with Ipomoea reniformis,
animals will be injected with subcutaneous doses of pilocarpine. Then the animals will
be placed individually in a large glass cylinder (height 30 cm, diameter 20 cm) at 21 ±
1oC and allowed to habituate for 15 minutes before injection of drugs. Numbers of
chews are counted by direct observation immediately after drug administration. The
results will be presented as numbers of chews in a 30 min period. 18
Anti-psychotic study using apomorphine induced climbing behavior
The albino wistar rat will be randomly distributed into four groups containing six
animals in each group. The groups will be treated with suitable doses of Ipomoea
reniformis or the standard. 60 minutes after the treatment with Ipomoea reniformis,
animals will be injected with haloperidol i.p. Then the animals are placed individually in
wire- mesh stick cages; 30 minutes after the treatment is given. 10, 20 and 30 minutes
after apomorphine administration, they are observed for climbing behavior and scored as
follows:
0 = four paws on the floor
1 = forefeet holding the vertical bars
2 = forefeet holding the bars
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7.5 Experimental design
Anti-epileptic activity (single and multiple dose study)
a. Maximal electro shock (MES) induced seizures
GROUP
TREATMENT AND
N=6
DOSE
I
Vehicle (1 ml/kg, p.o.) +
PARAMETERS
MES
II
Phenytoin (90 mg/kg, i.p.)
Hind limb tonic extension
+ MES
III
Ipomoea reniformis
(Lower) + MES
IV
Ipomoea reniformis
(Heigher) + MES
Multiple dose study is conducted for duration of 15 days.
b. Isoniazid (INH) induced seizures
GROUP
TREATMENT AND
N=6
DOSE
I
Vehicle (1 ml/kg, p.o.) +
PARAMETERS
INH
II
III
Diazepam (4 mg/kg, i.p.) +
Occurrence of tonic –
INH
clonic seizure
Ipomoea reniformis
(Lower) + INH
IV
Ipomoea reniformis
(Heigher) + INH
Multiple dose study is conducted for duration of 15 days. INH is administered on 15th
day.
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c. Pentylenetetrazol (PTZ) induced convulsions
GROUP
TREATMENT AND
N=6
DOSE
I
Vehicle (1 ml/kg, p.o.) +
PARAMETERS
PTZ
II
III
Diazepam (4 mg/kg, i.p.) +
Occurrence of tonic –
PTZ
clonic seizure
Ipomoea reniformis
(Lower) + PTZ
IV
Ipomoea reniformis
(Heigher) + PTZ
Multiple dose study is conducted for duration of 15 days. PTZ is administered on 15 th
day.
Antipsychotic activity (single and multiple dose study)
a. Apomorphine induced stereotype behavior
GROUP
TREATMENT AND
N=6
DOSE
I
Vehicle (1 ml/kg, p.o.) +
PARAMETERS
Apomorphine
II
Standard drug (1 mg/kg,
i.p.) + Apomorphine
III
Stereotyped behavior
Ipomoea reniformis
(Lower) + Apomorphine
IV
Ipomoea reniformis
(Heigher) + Apomorphine
Multiple dose study is conducted for duration of 15 days. Apomorphine is administered
on 15th day.
10
b. Pilocarpine induced purposeless chewing
GROUP
TREATMENT AND
N=6
DOSE
I
Vehicle (1 ml/kg, p.o.) +
PARAMETERS
Pilocarpine
II
III
Standard drug (1 mg/kg,
i.p.) + Pilocarpine
Numbers of chewing in 30
Ipomoea reniformis
minutes period
(Lower) + Pilocarpine
IV
Ipomoea reniformis
(Heigher) + Pilocarpine
Multiple dose study is conducted for duration of 15 days. Pilocarpine is administered on
15th day.
c. Apomorphine induced climbing behavior
GROUP
TREATMENT AND
N=6
DOSE
I
Vehicle (1 ml/kg, p.o.) +
PARAMETERS
Apomorphine
II
Standard drug (1 mg/kg,
i.p.) + Apomorphine
III
Climbing behavior
Ipomoea reniformis
(Lower) + Apomorphine
IV
Ipomoea reniformis
(Heigher) + Apomorphine
Multiple dose study is conducted for duration of 15 days. Apomorphine is administered
on 15th day.
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7.6 Does the study require any investigation or interventions to be conducted on
patients or other humans/animals? If so please describe briefly.
Yes, the above study requires investigation to be done on the adult albino wistar rats of
either sex for the anti-epileptic and anti-psychotic study.
7.7 HAS ANIMAL ETHICAL COMMITTEE CLEARNACE BEEN OBTAINED
FROM YOUR INSTITUTION IN CASE?
The study has been referred to the animal ethical committee of the institution for the
clearance. (Approval no:SSCPT/IAEC.clear/114/11-12.Dated:30-11-11)
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8
REFERENCES
1. Pandeya SN. A textbook of Medicinal Chemistry.3rd ed.SG Publisher Varanasi
158.
2.
Anonymous. Neurological disorders: Public health challenges. Switzerland:
World Health Organization; 2006.
3. Goodwin FK. Rationale for using lithium in combination with other mood
stabilizers in the management of bipolar disorder. J Clin Psychiatry. 2003;64
Suppl 5:18-24. Review.
4. Seizures
and
Epilepsy:
Hope
through
Research.
Available
from:
http://www.ninds.nih.gov/disorders/epilepsy/epilepsy.html. date 25-11-11.
5. Reddy DS, Pharmacotherapy of catameninal epilepsy. Indian J pharmacol
2005;37:288-93.
6. Wang NN, Anuka Ja, Kwanashie HO, Gang SS, Auta A. Anti-seizure activity of
the aqueous leaf extract of solanam nigram Linn (solanaceae) in experimental
animals. Afr Health Sci 2008;8(2):74-9.
7. Leunson D.F. 1991 pharmacologic treatment of schizophrenia. Clini, Ther
16,326
8. Mental health as chronic disease.
Available from:
http://www.tsha.nhs.uk/strategic-framework/pirority/3/transforming-ling-termConditions-management/mental-illness/as-a-chronic-diseases. date 25-11-11.
9. Mehul KB, Kishore KD, Ajay KS. Ipomoea reniformis: A Scientific Review.
International J Pharmacy Pharm Sciences 2010;(2):(4).
10. Agarwal VS, Drug Plants of India, 1st ed., Vol. 1, Kalyani Publishers, New
Delhi, 1947; 440.
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11. Sudhavani V, Chinnikrishnaiah V, Raghu Moorthy V, Raghavendra HG,
Ranganayakulu D. Nephroprotective activity of Merremia emarginata burm
against cisplatin induced nephrotoxic rats. J Adv Drug Res 2010;(1):(1).
12. Sanja SD, Sheth NR, Joshi DM, Golwala DK, Patel Dhaval, Raval MK. Antiinflammatory Activity of Ipomoea reniformis Methanolic Extract. International J
pharmaceutical Sciences and Drug Res 2009;1(3):176-9.
13. Rajiv Gandhi G, Sasikumar P. Antidiabetic effect of Merremia emarginata
Burm.F. in streptozotocin induced diabetic rats. Asian Pacific J Tropical
Biomedicine(2012)1-6.
14. Achliya GS, Wadodkar SG, Dorle AK. Evaluation of CNS activity of brahmi
ghrita. Indian J Pharmacol. 2005; 37(1): 33-6.
15. Pinelli A, Trivulzio S, Zefinetti GC, Tofanetti O. Anti-convulsant effects by
reduced glutathione and related amino acids in rats treated with isoniazid. J
Toxicol 1988; 48:103-7.
16. Maheshwar HG, Despande SV, Pramaod HJ. Anti-convulsant activity of fruits of
terminalia chebulia retz. Against MES and PTZ induced seizures in rats. J Herb
Med Toxicol 2010; 4(2): 123-6.
17. Vogel HG, editor. Drug discovery and evaluation. 2nd ed. New York: Springer
Berlin Heidelberg. 2002. 488, 531-7.
18. Finn M, Jassen A, Baskin P, Salamone JD. Tremulous characteristics of the
vacuous jaw movements induced by pilocarpine and ventrolateral striatal
dopamine depletions. Pharmacol Biochem Behav 1995; 57(1): 243-9.
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9.
Signature of the candidate
10
Remarks of the Guide
The above information and literature has been extensively investigated and verified. The
present study will be carried out under my supervision and guidance.
11
11.1 Name and Designation of
Guide
Mrs. BABITHA S
M.Pharm., (Ph.D)
Lecturer
DEPARTMENT OF PHARMACOLOGY
11.2 Signature
11.3 Co-Guide ( If any)
N.A
11.4 Signature
N.A
11.5 Head of the Department
Dr. THIPPESWAMY B.S.,
M. Pharm., Ph. D.
Professor & Head
DEPARTMENT OF PHARMACOLOGY
11.6 Signature
12
12.1 Remarks of the Chairman
and Principal
The above mentioned information is correct and I
recommend the same for approval.
12.2 Signature
Dr. S. Badami M.Pharm., Ph.D.
Principal
Sree Siddaganga College of Pharmacy
B. H. Road, Tumkur-572 102.
15