M. pharm dissertation protocol submitted to the rajiv gandhi university of health

M. PHARM DISSERTATION PROTOCOL

SUBMITTED TO THE

RAJIV GANDHI UNIVERSITY OF HEALTH

SCIENCES, KARNATAKA, BANGALURU.

BY

B. Pharm.

UNDER THE GUIDANCE OF

M. Pharm.,

P. G. DEPARTMENT OF PHARMACOLOGY

HARAPANAHALLI-583131

Rajiv Gandhi University of Health Sciences, Karnataka, Bangaluru

Annexure – II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

ENCLOSURE-I

Diabetes is a group of metabolic diseases characterized by hyperglycaemia resulting from defects in insulin secretion, insulin action, or both. The total number of people with diabetes is projected to explode from 171 million in 2000 to 366 million in 2030. Diabetes is usually associated with hypertension, poor metabolic control, smoking and obesity. Modern medicines like biguanides, sulphonylureas and thiozolidinediones are available for the treatment of diabetes. But they also have undesired effects associated with their uses. Alternative medicines particularly herbal medicines are available for the treatment of diabetes. Common advantages of herbal medicines are effectiveness, safety, affordability and acceptability. Medicinal plants and their products have been used in the Indian traditional system of medicine and have shown experimental or clinical anti-diabetic activity. Medicinal plants are a rich source of natural products. Medicinal plants and their products have been widely used for treatment of diabetic populace all around the world with less known scientific basis of their functioning. Hence, natural products from medicinal plants need to be investigated by scientific methods for their anti-diabetic activity¹.

Wound healing is the body’s natural process of regenerating dermal and epidermal tissue. The healing cascade is activated when platelets come into contact with exposed collagen leading to platelet aggregation and the release of clotting factors resulting in the deposition of a fibrin clot at the site of injury. The fibrin clot serves as a provisional matrix and sets the stage for the subsequent events of healing. Inflammatory cells also arrive along with the platelets at the injury site providing key signals known as growth factors. The fibroblast is the connective tissue cell responsible for collagen deposition required to repair the tissue injury. Collagen accounts for 30% of the total protein in the human body. In normal tissues, collagen provides strength, integrity and structure. When tissues are disrupted following injury, collagen is required to repair and restore normal structure and function².

The plant Leonotis nepetaefolia R.Br has medicinal application like hypoglycemic property and in management of painful, arthritic and other inflammatory conditions³. The plant is also used in the treatment of rheumatism, rickets, headaches and for the treatment of wounds⁴. However, antidiabetic and wound healing activities of title herb have not been documented in the scientific literature. In view of this, the present study is undertaken.

ENCLOSURE-II

The Leonotis nepetaefolia R. Br. is a erect annual herb belonging to the family; Lamiaceae (Mint family) commonly called lion’s ear or klip dagga. This plant commonly grown in Tropical Asia, Africa, Southern India and naturalized.

The plant Leonotis nepetaefolia R. Br. rich in alkaloids (leonurine and stachydrene), iridoid glycosides (leonuride, leonurin and leonuridine), diterpenoids (leocardin), flavonoids (rutin, quercetin, hyperoside, apigenin), volatile oil, tannins, vitamin A.

The root contains n-octacosanol, n-octacosanoic acid, quercetin, crumarin like 4,6,7-trimethoxy-5-methylchromene-2-one, campesterol and beta-sitosterol-beta-D-glucopyranoside. The seed oil contains oleic, linoleic, palmitic and stearic acids. The fatty oil, extracted from the seeds, is similar to olive oil. The leaves contain neptaefolin, neptaefuran, neptaefuranol, neptaefolinol, leonitin, neptaefolinin and (−)-55, 6-octadecadienoic acid.

Therapeutic uses:^3-10

The leaves of Leonotis nepetaefolia R.Br. are used traditionally in the management of type-2 diabetes mellitus, pain, arthritis, coughs, indigestion, warm infestation, nasal-disorders, skin infections, malaria and flowers are used in burns, scalds, itches and skin diseases . In the Carribean region plant was found to be used in treatment of asthma. The plant is also medicinally useful to treat rheumatism, rickets, headaches, wounds, ringworm, laxative, bitter, narcotic, fever and convulsion.

1. 
   This study is planned to investigate and validate the anti-diabetic activity against alloxan indused diabetic rat model and also wound healing activity will be assessed.
   
2. 
   In view of this plant Leonotis nepetaefolia R. Br have been selected for the present investigation.
   

ENCLOSURE – IV

Whole work is aimed to generate data from the laboratory that is experiments on animals. Albino rats and mice will be used for this purpose.

The experiments which involves.

• 
  Preparation of 70% hydroalcoholic extract of dried leaves of Leonotis nepetaefolia R. Br.
  
• 
  The preliminary phytochemical constituents present in the leaf extract will be carried out by following literature reported chemical tests.
  
• 
  The acute toxicity of extracts will be studied on mice following fixed dose method of CPCSEA
  
• 
  Crude extract of Leonotis nepetaefolia R. Br. leaves will be screened for anti-diabetic activity against alloxan induced diabetic rat model.^11-13
  
• 
  Wound healing activity of test extract will be screened using the following experimental animal model.¹⁴
  

For the present investigation, plant leaves of Leonotis nepetaefolia R. Br. will be collected from the surroundings of Harapanahalli. The plant will be cleaned and dried at room temperature, coarsely powdered and extracted. The crude extract thus obtained after removal of solvent will be subjected to preliminary phyto-chemical screening.

For this purpose female albino mice (20-25g) will be used. Fixed dose method (OECD guideline no. 420) of CPCSEA will be adopted.

(A)Estimation of the following biochemical parameters:

a) Glucose

b) Urea

d) Cholesterol

e) Protein

(B) Morphological studies:

-Recording body weight of animals.

(C) Histopathological examination of pancreas.

3. 
   Wound healing activity of test extract will be screened using the following experimental animal model.
   

Excision wound model.

1. 
   Percentage of wound contraction.
   
2. 
   Period of epithelization.
   

The results obtained from the above investigation will be subjected to statistical analysis using one way ANOVA followed by Tukey- Kramer Multiple Comparision test.

Total duration for the completion of proposed research work may be ten months

1. Collection of plant material. - One month.
2. Duration of experimentation on animals including - Five months.

preparation of crude extracts.

4. Dissertation writing and communication of research - Two months.

papers.


ENCLOSURE – VI

7.3 Does the study require any investigation or interventions to be conducted on patients or other humans or animals? If so, please describe briefly.

Albino rats of Wistar strain will be used for the evaluation of anti-diabetic and wound healing properties. Whereas albino mice will be used for acute toxicity study.

4. 
   Has ethical clearance been obtained from your institution in case of 7.3?
   

ENCLOSURE – VII

8. 
   List of references:
   

1. 
   Dineshkumar B, Mitra A, Manjunatha M. Studies on the antidiabetic and hyupolipidemic potentials of mangiferin (xanthone glucoside) in streptozotocin-induced type 1 and type 2 diabetic model rats. International Journal of Advances in Pharmaceutical Sciences 2010; 1: 75-85.
   

2. 
   Nayak BS, Raju SS, Ramsubhag A. Investigation of wound healing activity of Lantana camara L. in Sprague dawley rats using a burn wound model. International Journal of Applied Research in Natural Products 2008; 1(1): 15-19.
   

3. 
   http://www.tropilab.com/lionsear.html
   

4. 
   Ayanwuyi LO.Yaro AH, Adamu HYS. Studies on anticonvulsant activity of methanol capitulum extract of Leonotis nepetifolia Linn. Nigerian Journal of Pharmaceutical Sciences 2009; 8(1): 73-79.
   

5. 
   Kirtikar and Basu, Indian Medicinal Plants 2^nd edition, vol 3;p-2023-25.
   

6. 
   Chetty KM, Sivaji K, Rao KT. Flowering plants of chitoor district Andrapradesh India, student offset printers Tirupathi. P- 276 .
   

7. 
   http://www.floridata.com/ref/l/leon_nep.cfm
   

8. 
   Khare CP. Indian Medicinal Plants An Illustrated Dictionary. Springer 2007: p-368.
   

9. 
   Rastogi RP, Mehrotra BN. Compendium of Indian Medicinal Plants 1970-1979. Central Drug Research Institute, Lucknow and National Institute of Science Communication And Information Resource, New Dehli 2006; 2: p-411.
   

10. 
    Chopra RN, Nayr LS, Chopra CI, Glossary of Indian Medicinal Plants; p-152.
    

11. 
    Srinivasan S, Karundevi B, Comparative evaluation of hypoglycaemic activity of two medicinal plants in alloxan diabetic rats. International Journal of Pharmacology 2005, 1(3); p-267.
    

12. 
    Trivedi NA, Mazumdar B, Bhatt ID, Hemavathi KG. Effect of shilajith on blood glucose and lipid profile in alloxan-induced diabetic rats. Indian J Pharmacology. Dec2004; 36; p-373.
    

13. 
    Senthil Kumar GP, Arulselvan P, Sathish Kumar D and Subramanian SP. Antidiabetic activity of fruits of Terminalia chebula on streptozotocin indused diabetis in rats J of Health Science 2006;56(3); p-283.
    

14. 
    Shanbhag TV, Sharma chandrakala, Sachidananda A, Laxminarayana kurady B, Shenoy Smita and Shenoy Ganesh. Wound healing activity of alcoholic extract of Kaempferia galanga in wistar rats. Indian J Physiol Pharmacol 2006; 50(4); p-384.
    

15. 
    Prema Veeraraghavan. Expert consultant, CPCSEA, OECD Guideline no.2000; p- 420.