FORMULATION AND EVALUATION OF TOPICAL LIPOSOMES WITH AN ANTIFUNGAL DRUG
M. Pharm. Dissertation Protocol Submitted to
Rajiv Gandhi University of Health Sciences, Karnataka
Bangalore– 560041
By
MISS. ASHWINI. ALUR B.Pharm
Under the Guidance of
Mr. SHALIN .P. THAKKER M.Pharm
Professor
Department of Pharmaceutics
S.E.T’s COLLEGE OF PHARMACY
S. R. Nagar, Dharwad–580002
2012-2013
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,
BANGALORE, KARNATAKA
ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1. / NAME OF THE CANDIDATE AND ADDRESS / MISS. ASHWINI ALURSET’s COLLEGE OF PHARMACY,
S. R. NAGAR,
DHARWAD-580002
2. / NAME OF THE INSTITUTION / SET’s COLLEGE OF PHARMACY,
S. R. NAGAR,
DHARWAD-580002
3. / COURSE OF STUDY AND SUBJECT / MASTER OF PHARMACY IN PHARMACEUTICS
4. / DATE OF ADMISSION TO COURSE / JUNE- 2011
5. / TITLE OF THE TOPIC
“FORMULATION AND EVALUATION OF TOPICAL LIPOSOMES WITH AN ANTIFUNGAL DRUG”
BRIEF RESUME OF THE INTENDED WORK
6.1 NEED FOR THE STUDY
Fungal infections are caused by the various species of fungi. These fungi are also known as Dermatophytes. Dermatophytes are capable of causing superficial skin infections known as Dermatophytosis. The causative fungi belongs to three genera namely Microsporum, Trichophyton and Epidermophyton. Clinical forms of cutaneous infection includes Tinea Corporis ( involvement of body), Tinea Capitis (scalp involvement), Tinea Pedis (feet involvement) and Onychomycosis (nail involvement).1
Antifungal drugs are used for superficial and deep (systemic) fungal infections. Clotrimazole and Econozole belongs to Imidazole derivative where as Fluconozole and Itraconozole belongs to Triazoles derivative. Imidazoles are used for topical purpose. Triazoles are used for both topical and systemic purpose. The mechanism of action of Imidazoles and Triazoles is same (Imidazoles and Triazoles acts by inhibiting fungal cytochrome P450 enzyme “lanosterol 14-demethylase” which impairs ergosterol synthesis leading to cascade of membrane abnormalities in the fungus.)2
Econozole or Itraconozole acts by inhibiting fungal cytochrome P450 enzyme “lanosterol 14-demethylase” which impairs ergosterol synthesis leading to cascade of membrane abnormalities in the fungus.2
Depending upon the infection rate these drugs are administered in two different forms i.e topical form and systemic (injectables and oral) form. Econozole or Itraconozol can be administered as:
1. Topical Ointment.
2. Topical Gels.
3. Topical Liposomes.
4. Topical Emulgel.
Liposomes are microscopic bi-layered vescicles in which an aqueous phase is entirely enclosed by a membranous lipid layer mainly composed of natural and synthetic phospholipids. Liposomes are formed when phospholipids are hydrated. Phospholipids are amphipathic in nature having both hydrophobic tail and hydrophilic or polar head.3 In aqueous medium the molecules are oriented in such a way that the polar portion of the molecule remains in contact with the polar environment and at the same time shields the non-polar part.3
Topical liposome form is the most suitable form of administration of drug. The entrapment of drug in liposomes can facilitate localized delivery of the drug. This improves availability by means of a sustained- release pattern of drug, thus it advances the treatment of deep fungal infections.4 As there is encapsulation of drug in liposomes there will be increase in drug efficacy, this inturn increases the stability of the dosage form and minimizes side effects5,9 because of this reason Liposomal drug delivery system
is better than any other conventional dosage form. Liposomes acts as vescicles that facilitate the transport of active drug through skin.14
6.2 REVIEW OF LITERATURE:
R. Agarwal and O.P.Katare prepared Miconozole nitrate-loaded topical liposomes thin film hydration method using Phosphatidyl choline, Cholesterol and Miconozole nitrate. Thin film is hydrated using 6.4 pH phosphate buffer for 2hrs. The prepared liposomes were characterized for specific properties such as microscopic appearance, size and degree of drug entrapment. Significant retention of Miconozole in skin was observed with the use of liposomes.4
Patel et al. formulated and evaluated ketoconozole liposomes. Antifungal drug, ketoconozole was encapsulated in liposomes for topical application. Ketoconozole liposomes were prepared by thin film hydration technique using soya lecithin, cholesterol and drug in different weight ratios.The prepared liposomes were characterized for size, shape, entrapment efficiency, and invitro drug release [by franz diffusion cell] using membrane. The studies were demonstrated successful preparation of Liposomes containing Ketoconozole. The effect of soya lecithin:cholesterol ration achieved maximum entrapment efficiency.5
Rakesh P.Patel et al. formulated and evaluated topical Carbopol gel formulation containing Ketoconozole encapsulated liposomes. Ketoconozole loaded liposomes were prepared by thin film hydration technique.The prepared liposomes were incorporated into 1% Carbopol gel and were evaluated for invitro drug release,drug retention into skin and antifungal activity. The release of Ketoconozole from liposomal gel was much slower in a controlled release pattern. Thus gel containing liposomal Ketoconozole showed maximum antifungal activity.6
M.A.Repka et al prepared a film containing antifungal drug, Ketoconozole for Onychomycosis using Hot-melt extrusion[HME] method to prepare films containing 20%w/w Ketoconozole. Ketoconozole 0.125%gel was prepared using Carbopol 974P. Films were processed at a temperature range of 115-120oC utilizing a killion extruder and were evaluated for post-extrusion drug content ,content uniformity ,bioadhesion, thermal behavior and nail drug permeation. The extruded films demonstrated good content uniformity and post processing drug content. Tensile and peel tests were recorded to determine the bioadhesive profiles.7
Ankur Jain, Piyusha Deveda, Naveen Vyas et al developed antifungal emulsion based gel for topical fungal infections. Stability of Emulsion increases when it is incorporated in gel. The objective of the project was to develop an emulsion based gel for control delivery for Miconozole to treat local as well as systemic fungal infections. In this they have prepared emulsion and incorporated in carbopol gel. The prepared formulation was evaluated on basis of pH, spreadability, viscosity, drug content, invitro release and stability. The results reveled that the batches shows 94.80% release in 24hrs and were stable for around three months.8
Mayank R.Joshi and A.N.Mishra prepared and optimized liposomes of Terbutaline sulfate [TER]. Liposomes were prepared using drug, lipid[CHOL] and phosphoytidyl choline[PC] by Lipid film hydration technique in a rotary flask evaporator.The prepared liposomes were characterized for size, shape and lamellarity. The stability of liposomes in terms of retention of TER was measured at refrigeration temperature [2-8oC], room temperature and oven temperature [37oC] for a period of 3months. The results suggested that the entrapment of TER in liposomes increased by increasing the proportion of PC and maximum encapsulation and retention of TER was achieved only with a specific PC:CHOL molar ratio.9
J N Khandare, Rashmi Dalmla and Rajesh Manchanda, evaluated Ketoprofen Liposomes for anti-arthritic activity. Here Ketoprofen was encapsulated in liposomes using different molar ratios of Lecithin and Cholesterol. Anti-arthritic activity of topically applied liposome encapsulated drug in FAPG[fatty alcohol propylene glycol] base was studied by formaldehyde induced arthritis method. Results indicated a higher anti-arthritic activity of liposomes of ketoprofen compared to plain drug.10
S.Agrawal et al prepared Liposomes of Daunorubicin for reduced Cardiotoxicity in the face of unaltered Antitumour activity in Swiss Mice bearing Fibrosarcoma. Here Liposomes were prepared by the
Versatile pH gradient method with an aim to modify drug disposition and increase the therapeutic efficacy of the drug, Daunorubicin. The preparation were then characterized with respect to size and its distribution, entrapment efficiency, invitro drug release profile and its stability under specified conditions of storage. Antitumour efficicacy bearing solid tumour namely Fibrosarcoma and organ toxicity studies in swiss albino mice were conducted with liposomes administered and the result was that drug entrapment efficiency was found to be 82.5±1.28 % with a drug being incorporated into the aqueous layer of the vescicles. The results revealed that encapsulation of drug in liposomes are a potential tool for the delivery of drug.11
Gita Rao and R S R Murthy evaluated Liposomal Clobetasol Propionate topical formulation for intra dermal delivery. Here they have prepared three formulations like Hydroxy propyl methyl cellulose[HPMC] K4M gels with only drug, liposome encapsulated clobetasol propionate and physical mixture of drug and lipids. These formulations were subjected to invitro drug diffusion studies using rat skin to determine diffusion parameters. Data were analysed to calculate the amount of drug in the skin. Invivo skin blanching assay of these formulations in human volunteers. Out of all formulations tested, liposomal gel prepared with drug, phosphotidyl choline and cholesterol in the ratio of 2:4:1 showed minimum blanching score.12
6.3 OBJECTIVE OF STUDY:
The objective of the proposed study are:-
§ To develop the topical liposome of an antifungal drug.
§ To evaluate formulations for various quality control parameters.
§ To compare the formulations made of various lipid bases and surfactants.
§ Evaluation of formulations for various characterization.
7. Materials and methods :
7.1 SOURCE OF DATA :
· Reference books.
· Web resources.
· Indian Journal of Pharmaceutical Sciences.
· International Journal of Pharma. Research and development.
· International Journal of Pharmaceutics.
· European Journal of Pharmaceutics and Biopharmaceutics.
· International Journal of Drug Delivery technology.
· Indian Drugs.
7.2 METHOD OF COLLECTION OF DATA :
A. materials :
Drug : Any of the Antifungal drug will be used.
Lipid bases: Cholesterol and stearic acid.
Surfactants: Soya lecithin and phosphatidyl choline.
Solvents : Chloroform.
Chemical and other reagents required for preparation of topical liposomes will be procured from standard company sources.
B. METHOD OF PREPRATION :
Thin film hydration technique using Rotary Evaporator. OR
Thin film hydration technique using Hand shaking method.5
C. EVALUATION PARAMETERS:
1. CHARACTERIZATION OF TOPICAL LIPOSOME OF ANTIFUNGAL DRUG:
Following parameters will be evaluated,
§ Particle size and size distribution.
§ Drug entrapment efficiency.
§ Differential scanning calorimetry (DSC) study.
§ Scanning electron microscopy.
§ Measurement of pH.8
§ Invitro drug release :
In-vitro drug release will be carried out using Franz diffusion cell as diffusion test apparatus. The diffusion medium consists of buffer with pH 5.5.9 A 50ml of the dissolution fluid is used with stirring speed of 50rpm at 37± 0.5oC.12 Samples of specified volume are withdrawn at suitable time intervals by replacing the same dissolution medium and samples are analysed by measuring the absorbance using UVspectrophotometer.5§ Microbiological assay
7.3 DOES THE STUDY REQUIRE ANY INVESTIGATION OR INVENTION TO BECONDUCTED ON PATIENTS OR OTHER HUMANS OR ANIMALS? IF SO
PLEASE MENTION BRIEFLY.
No.
7.4 HAS ETHICAL CLEARANCE BEEN OBTAINED FROM YOUR
INSTITUTION IN CASE OF 7.3?
Not applicable.
REFERENCES:
1. Colledge R N, Walker R B,Ralston H S. Davidson’s Principles and Practices of Medicine. 2010. 21st edition.p.1273.`
2. Tripathi K D. Essentials of Medical Pharmacology. 6thedition. New Delhi: Jaypee Brothers Medical Publishers: 2008.p.757-65.
3. Vyas S P, Khar R K, Targetted and controlled drug delivery, Novel Carrier System CBS Publishers and distributors: 2010.p.173-248
4. Agarwal R, Katare O P, Preparation and Invitro evaluation of Miconozole nitrate- Loaded Topical Liposomes. Pharm Tec 2002:48-60.
5. Patel R P, Patel H H, Baria A H. Formulation and Evaluation of Liposomes of Ketoconozole. Int. J Drug Delivery Tech 2009;1(1):16-23.
6. Patel R P, Patel H H, Baria A H. Formulation and Evaluation of Carbopol gel containing Liposomes of Ketoconozole. Int. J Drug delv tech 2009;1(2):42-45.
7. Repka M A, Mididoddi K P, Stodghill P S. Influence of nail etching for assessment of topical
Onychomycsis therapies. Int. J Pharm 2004;282:95-106.
8. Jain A, Deveda P, Vyas N, Chauhan J, Khambete H, Jain S. Development of antifungal emulsion based gel for topical fungal infections. Int J Pharm Res Development. 2011;2(12):18-25.
9. Joshi R M, Mishra N A. Liposomes of Terbutaline Sulfate. Preparation, Optimisation and Stability studies. Indian drugs 1999 Apr; 36(4): 245-52.
10. J N Khandare, Dalmla R, Manchada R. Evaluation of Ketoprofen Liposomes for the Anti-Arthritic activity. Indian Drugs 1995Sep; 33(2):63-66.
11. Agarwal S, Tiwari S, Udupa N et al. Liposomal Daunorubicin: Reduced Cardiotoxicity in the face of unaltered Anti tumour activity in swiss mice bearing Fibrosarcoma. Ind J Pharm Sci 2002; 64(1):18-23.
12. Rao G, Murthy R S R. Evaluation of Liposomal Clobetasol Propionate Topical formulation for intra dermal delivery. Ind J Pharm Sci 2000:459-62.
13. Sant V P, Paradkar A R, Nagarshekar M S. Optimisation of Pentoxifylline Liposomes using 24 Factorial design. Ind J Pharm Sci 2002; 64(5):459-64.
14. Sood A,Venugopalan P, Venkateshan N, Vyas S P. Liposomes in cosmetics and skin care; Indian Drugs 1995 Oct 19; 33(2).
15. Pattan S R, Chavan S S, Jadhav R S, Nirmal S A, Tambe V D. Liposomes in targeted drug delivery. Indian Drugs; 2011 Aug; 48(08):5-13.
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9. / SIGNATURE OF THE STUDENT10. / REMARK OF THE GUIDE:
The above mentioned information and literature has been extensively investigated, verified and was found to be correct. The present study will be carried out under my supervision and guidance.
11. / 11.1 NAME AND DESIGNATION
OF THE GUIDE
11.2 SIGNATURE / Prof. S.P. THAKKER M. Pharm.
PROFESSOR AND HEAD,
DEPT. OF PHARMACEUTICS
SET’s COLLEGE OF PHARMACY, S.R.NAGAR, DHARWAD- 580002.
11.3 NAME AND DESIGNATION
OF CO-GUIDE
11.4 SIGNATURE / ------
11.5 HEAD OF THE
DEPARTMENT
11.6 SIGNATURE / Prof. S.P. THAKKER M. Pharm.
PROFESSOR AND HEAD,
DEPT. OF PHARMACEUTICS
SET’s COLLEGE OF PHARMACY, S.R.NAGAR, DHARWAD- 580002.
12. / 12.1 REMARK OF THE
PRINCIPAL
12.2 SIGNATURE / The above mentioned information is correct and I recommend the same for approval.
Dr. V. H. KULKARNI M. Pharm, Ph.D.
PROFESSOR & PRINCIPAL,
SET’s College of Pharmacy,
S.R.NAGAR, DHARWAD- 580002.
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