Formulation and Evaluationof Gel Containing Proliposomes for Transdermal Delivery of Metformin

FORMULATION AND EVALUATIONOF GEL CONTAINING PROLIPOSOMES FOR TRANSDERMAL DELIVERY OF METFORMIN HYDROCHLORIDE

Synopsis for M.Pharm Dissertation submitted to the

RajivGandhiUniversity of Health Sciences Karnataka, Bangalore.

By

Ms.SHRUTHI M V

M.Pharm. Part-I

Under the guidance of

Mr.S.PARTHIBAN., M Pharm.,

Department of Pharmaceutics,

Bharathi College of Pharmacy,

Bharathinagara.

2013-2014

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

KARNATAKA, BANGALORE

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. / NAME AND ADDRESS OF THE CANDIDATE / SHRUTHI M V
M.PHARM., PART-I,
DEPARTMENT OF PHARMACEUTICS,
BHARATHI COLLEGE OF PHARMACY,
BHARATHI NAGARA, MANDYA,
KARNATAKA-571422.
2. /

NAME OF THE INSTITUTION

/ BHARATHI COLLEGE OF PHARMACY,
BHARATHI NAGARA.
3. /

COURSE OF STUDY AND SUBJECT

/ MASTER OF PHARMACY IN PHARMACEUTICS.
4. / DATE OF ADMISSION OF COURSE / 15.01.2013
5. / TITLE OF TOPIC / FORMULATION AND EVALUATIONOF GEL CONTAINING PROLIPOSOMES FOR TRANSDERMAL DELIVERY OF METFORMIN HYDROCHLORIDE.
6. / BRIEF RESUME OF THEINTENDED WORK
6.1 Need for the study
6.2 Review of the literature
6.3 Objectives of the study / ENCLOSURE - I
ENCLOSURE - II
ENCLOSURE – III
7. /

MATERIALS AND METHODS

7.1 Source of data

7.2 Method of collection of data

7.3Does study requires any investigation or interventions to be conducted on patients or other human or animal? If so, please describe briefly.

7.4 Has ethical clearance been obtained from your institution in case of 7.3

/ ENCLOSURE - IV
ENCLOSURE - V
ENCLOSURE - VI
ENCLOSURE-VI
8. / LIST OF REFERENCES / ENCLOSURE – VII
6.0
7.0 / Brief resume of the intended work:
ENCLOSURE – I
6.1 – Need for the study
Liposomes are the most promising and broadly applicable of all the novel delivery systems. For liposomes to enter the market, they must be stable during the storage period, and remain intact before reaching their targeted tissues to produce action. Various approaches have been used to overcome these problems, some of which include, control of particle size and lamellarity, altering the lipid composition,lyophilisation,electrosteric stabilization etc. One of such approach which helped to overcome the stability issue associated with liposome and led to the development of a new drug delivery system is the Proliposome (PL) discovered by Payne et al., in 1986. Proliposomes (PLs) are dry, free-flowing granular products composed of drug(s) and phospholipid(s) which, upon addition of water, disperse to form a multi-lamellar liposomal suspension. It is one of the most cost-effective and widely used methods for producing commercial liposome products. It is based upon the intrinsic property of hydrated membrane lipids to form vesicles on contact with water. Being available in dry powder form, they are easy to distribute, transfer, measure and making it a versatile system. Liposomes can either be formed in vivo under the influence of physiological fluids or can be formed in vitro prior to administration using a suitable hydrating fluid. The liposomes formed on reconstitution are similar to conventional liposomes and more uniform in size.1Morever, by controlling the size of the porous power in proliposomes, relatively narrow range of reconstituted liposome size can be obtained. Because of these properties, proliposomes appear to be an elegant alternative to liposome in design and fabrication of liposomal dosage forms.
When proliposomes are applied to mucosal membranes, they are expected to form liposome upon hydration by mucosal fluids. The resulting liposome may act as a sustainedrelease dosage form of loaded drugs. As a matter of fact, blood concentration of propranolol could be sustained substantially by intranasal administration of a proliposomal powder. Stimulated by these findings, we wished to extend the application of proliposomes to systemic delivery of the drugs across the skin. This would be possible if proliposomes form liposomes upon hydration by the sweat on the skin following topical application under occlusive condition.2
Diabetes mellitus isgroup of metabolic disorders characterized by chronic hyperglycemia.3 In recent years, developing nations have witnessed an explosive increase in the prevalence of diabetes mellitus predominantly related to life science changes and the resulting surge in obesity. The metabolic consequences of prolonged hyperglycemia and dyslipidemia including accelerated atherosclerosis, chronic kidney disease pose enormous burden of patients with diabetes mellitus and on the public health system. An estimated 20.8 million people currently has diabetes, of these, 6.2 million or about 1/3rd.were undiagnosed. In 2005 alone, over 1.5 million new cases in adults were diagnosed. Globally the prevalence of diabetes for all ages is estimated to be 2.8% in 2000 and projected to 4.4% by 2030. The centers for disease control and prevention predicts the national incidence of diabetes will rise by 37.5% by the year 2025.4
Metformin hydrochloride, an oral anti-diabetic drug frequently used as first line drug of choice in treatment of type 2 diabetes, particularly in overweight and obese people and those with normal kidney function. Metformin hydrochloride is anti-hyperglycemic drug and it does not increase insulin release in the pancreas. Metformin hydrochloride reduces glucose levels primarily by decreasing hepatic glucose production and by increasing insulin action in muscle and fat. Metformin hydrochloride is absorbed mainly from the small intestine and does not bind to plasma proteins. Metformin hydrochloride is safe and not teratogenic in many of species studied. Metformin hydrochloride however has many gastro intestinal side effects including nausea, vomiting, anorexia and diarrhoea and rapid first pass metabolism.5 The transdermal Metformin hydrochloride allows delivery of drug through the skin and bypassing the digestive system.
This route has many advantages from orally dosed metformin;
They are,
1. Avoids gastrointestinal side effects by avoiding the digestive system.
2. Potentially reduces cost through fever side effect.
3. Smaller dose is required when metformin in penetrated through skin.
4. Improve the bioavailability as well improve the patient compliance.
Hence, in the present investigation, an attempt is made to formulate Metformin hydrochloride proliposomal gel in order to increase bioavailability and reduce side effects by achieving transdermal drug delivery.
ENCLOSURE - II
6.2REVIEW OF LITERATURE

Jessy Shaji et al., reviewed the method of preparation and evaluation of proliposome and highlights its potential to be exploited for different routes of administration.They reported that the poor stability associated with liposomeon long term storage can be overcome by the formulation of proliposomes with ability to deliver the drug in controlled release.1
Bo-Young Hwang et al., investigated proliposomes containing Nicotine by a standard method using sorbitol and lecithin. They have reported that sustained delivery of nicotine across the skin appears to be achievable through topical application of Nicotine-loaded proliposomes under occlusive conditions.2
Prakash V Diwan et al., studied on proliposomal gel bearing a steroidal anti-inflammatory agent Prednisolone intended for topical application. They prepared proliposomes by thin film hydration technique by using varying the lipid phase composition (lecithin/cholesterol) andreported that Prednisolone proliposomal gel showed sustain release with enhanced anti-inflammatory activity implicating its potential in effective topical pharmacotherapy for the treatment of rheumatoid arthritis.6
Gupta V et al.,developed and characterized a vesicular drug carrier system (proliposomes) for topical delivery of Aceclofenac to overcome the problems related with oral route. Aceclofenac proliposomes were prepared by film deposition on carrier method. They have reported in vitro release of drug was significantly retarded indicating sustained release of Aceclofenac from proliposomes with improved stability.7
Kurakula et al.,studied on proliposomal gel bearing a non-steroidal anti-inflammatory agent, Piroxicam intended for topical application. Proliposome formulations were prepared by thin film hydration technique using varying the lipid phase composition (lecithin/ cholesterol).They reported that the Piroxicam proliposomal gel showed sustain release with enhanced anti-inflammatory activity implicating its potential in effective topical pharmacotherapy for the treatment of rheumatoid arthritis.8

Byung-Nak Ahnet al.,preparedproliposomes by the penetration of a methanol-chloroform solution of Propranolol hydrochloride (PH)and lecithin into microporous sorbitol, with subsequent vacuum drying. They concluded that granular proliposomes of PH with good flowability and sustained release characteristics could be prepared by controlling the drug/lecithin/sorbitol ratio and sorbitol particle size and PH proliposomes can be potential candidates for the sustained drug delivery of propranolol when applied directly onto the mucosal membranes.9
Ning M Y et al., prepared Clotrimazole (CT)-containing proliposomes by penetrating an ethanol solution of CT and Egg phosphatidylcholine (PC) into micro porous sorbitol particles, followed by vacuum evaporation of the solvent. Their study shows that CT-containing vaginal proliposomes prolonged drug release and may increase amount of drug retention into the mucosa to result in more antifungal efficacy.10
Wipaporn Rojanaratet al., developed Levofloxacinproliposomes in a dry powder aerosol form for pulmonary delivery by spray drying technique usingsoybean phosphatidylcholine, cholesterol and porous mannitol. They have reported Levofloxacin and Levofloxacin proliposomes were shown to be nontoxic to respiratory-associated cells, and also did not activate AMs toproduce inflammatory cytokines and nitric oxide at a level that would cascade to show secondary inflammation.11
Sandeep loona et al., investigated a proniosomal carrier system of Metformin hydrochloride for the treatment of type - 2diabetes mellitus that is capable of delivering entrapped drug over an extended period of time.They have reported that the Metformin proniosomal gel is promising prolonged drug delivery system and has reasonably good stability characteristic.12
Brocks et al.,developed a proliposomal formulation to increase the oral bioavailability of Halofantrine [HLF] enantiomers, a drug with low and erratic oral bioavailability.13
Madhvi M et al.,reportedMetforminnoisome drug delivery system and evaluate its in‐vitro performance. The formulations were prepared with different types of surfactant.The study is based on formulations of Metformin noisome which provide most promising oralbioavailability of Metformin.14
Chuandi Sun et al., investigated the possibility of liquid proliposomes being carriers for oral delivery. They prepared Nimodipine liquid proliposomes-based soft capsules (NPSC) and reported that proliposomes shows potential way to improve oral delivery of Nimodipine.15
Rajesh Kumar et al., formulatedproliposomes in the form of enteric-coated beads using Glyburide as a model drug. The beads were enteric coated with Eudragit L-100 by a fluidized bed coating process using triethyl citrate as plasticizer.The dissolution study of enteric-coated beads exhibited enhanced dissolution compared with pure drug and a marketed product.16
Suman Ramteke et al., developed and characterized microemulsion for topical delivery of Glipizide. They are prepared by water titration method using oleic acid as oil phase, tween-80 as surfactant and propylene glycol as co-surfactant. They have reportedthat the Glipizide based microemulsion delivered the drug in sustained or controlled manner and prolonged delivery as compared to conventional dosage form.17
Sharma Dinesh Kumar et al.,prepared solid dispersed Glibenclamide using pregelatinesed starch by physical and kneading method. The prepared formulation shows enhanced dissolution and hypoglycemic activity.18
Deepthi Anna Kulaet al.,reviewed the drug delivery systems using colloidal particulate carriers such as liposomes and noisome have distinct advantages over conventional dosage forms and also having significant problems like instability. To overcome this problem provesicular concept has evolved to resolve the stability issues pertaining to the conventional vesicular systems. The new emerging concept has demonstrated the potential ofproliposomes/proniosomes in improving the oral bioavailability and permeation of drugs acrossthe stratum corneum.19
Jin-Ming Li et al.,developed a stealthy Etoposide proliposomes and studied the pharmacokinetics inrabbits. Blankstealthy liposomes were prepared by film dispersion method. Encapsulation efficiency of stealthy etoposideproliposomes was determined by Sephadex chromatography. The prepared stealthy Etoposide proliposomes could significantly extend the duration of etoposide in blood circulation.20

ENCLOSURE – III
6.3 OBJECTIVE OF STUDY
The specific objective of the study is toformulate and evaluate the proliposomal gelof Metformin hydrochloride for transdermal drug delivery.

To formulateproliposomes of Metformin HCl using various drug-excipient ratio.
To characterize the above formulations using various parameters which includes vesicular shape and surface morphology, vesicular size and size distribution, entrapment efficiency etc.,
To prepare proliposomal gel using carbopol gel base.
To evaluate the prepared proliposomlgelfor different parameter.
To perform the hypoglycemic activity using rats as an animal model.
To carry out stability studies on the selected formulation as per ICH guidelines.

MATERIALS AND METHODS
MODEL DRUG: Metformin hydrochloride
POLYMERS:Carbopol (or) any other suitable polymer will be selected.
Excipients:Glycerin, Lecithin, Cholesterol, Acetonitrile, Triethylamine, Mannitol,
Ethanol etc.,
METHOD:

Thin film deposition on carrier methodby using vacuum rotary evaporator.

Evaluation:
Compatibility study by Fourier Transform Infra-Red (FTIR) studies.
Surface morphology by scanning electron microscope (SEM)/ (TEM).
Vesicular size and count.
Zeta potential.
Drug entrapment efficiency.
Content uniformity of gel.
In-vitro diffusion studies by Franz diffusion cell.
Skin irritation test.
Perform the hypoglycemic activity using rats as an animal model.
Stability study.
ENCLOSURE - IV
7.1 Source of Data:

From Science Direct & other internet facilities.
Presentations like pharmaceutical poster presentation.
Research publications.
International and Indian journals.
Textbooks and reference books.
RGUHS Library.

JOURNALS:
Journals & articles :
International Journal of Pharma and Bio Sciences
Indian Journal of Pharmaceutical Education and Research
International Journal of Pharma Research and Review
International Journal of Pharmacy and Pharmaceutical Sciences
International Journal of Pharmaceutical Sciences and Drug Research
International Journal of Research in Pharmaceutical and Biomedical science
Indian Journal of Pharmaceutical Science
Journal of Controlled Release
Archive of Applied Science Research
Journal of Microencapsulation
International Research Journal of Pharmacy
RELATED LINKS:

ENCLOSURE –V
7.2Method of collection of data:
PART-I:
Extensive literature survey.
Procurement of raw materials and drug.
Standardization of raw materials and drugs.
PART-II:
Drug-excipient compatibility can be confirmed by carrying out FTIR studies.
To formulate proliposomes of Metformin hydrochloride using vacuum rotary evaporator.
To prepareproliposomalgel of Metformin hydrochloride using carbopol gel base.
PART-III: Evaluation of proliposomal gel:
Evaluation of the formulated proliposomal gel for different parameter such as

Physical examination
pH optimization
Drug content
Rheology
Stability studies
In vitro diffusion studies
In vivo studies

ENCLOSURE VI
7.3- Does the study require any investigations or interventions to be conducted on
patients or other humans or animals? If so, Please describe briefly.
-YES-
A.Skin irritation studies:
Three young rats of white strain will be taken for skin irritation studies. The test will be carried out using drug free polymeric films as control and proliposomal gel containing drug for the observation of erythema.
B.Hypoglycemicactivity:
Wisteralbino rats of either sex weighing 100-150g will be selected for in vivo studies. The reduction in blood glucose level will be observed in diabetes induced rats using alloxan monohydrate by comparing oral Metformin hydrochloride as standard with optimized formulation. . The blood is to be withdrawn by pricking the rat’s tail and blood glucose level will be measured by using digital glucometer (Accque check).
7.4 - Has ethical clearance been obtained from your Institution in case of 7.3? The study is cleared from ethical committee of the institution.
(Reg.No.01/CEU/IAEC/2013-2014 Dated 02/09/2013)
8.0 / ENCLOSURE - VII
LIST OF REFERENCE:

Jessy Shaji and Vinay Bhatia. Proliposomes: A brief overview of novel delivery system. Int Pharm Bio Sci,2013;4(1):150-160.
Bo-Young Hwang, Byung-Hwa Jung, Suk-Jae Chung, Min-Hwa Lee et al.,In vitro skin permeation of Nicotine from proliposomes. J Control Release, 1997;49:177-184.
Sujatha S,Swaroopa Rani V and Ravi Kumar B. Antidiabetic effect of flower extract of antigonon leptopus hook and are in alloxan-induced diabetic rats. Indian J Pharm Educ Res,2012;46(1):9-16.
Ravi Teja Allena, Hemant K S Yadav, Swetha Sandina, Sarat Chandra Prasad M. Preparation and evaluation of transdermal patches of Metformin hydrochloride using natural polymer for sustained release. Int J Pharm pharm Sci,2012;4(l3):297-302.
Bhavesh D, Chetan G, Bhat K M, Shivprakash.Estimation and pharmacokinetic of Metformin in human volunteers. Indian J Pharm Educ Res,2007;41(2):135-139.
Mallesh Kurakula, Srinivas C, Nagasree Kasturi, Prakash V Diwan. Formulation and Evaluation of Prednisolone Proliposomal Gel for Effective Topical Pharmacotherapy. IJPSDR,2012;4(1):35-43.
Gupta V, Barupal A K, Ramteke S. Formulation development and in vitro characterization of proliposomes for topical delivery of Aceclofenac. Indian J Pharm Sci,2008;70:768-78.
Kurakula, Mallesh, pasula, Nikitha et al., Piroxicam proliposomal gel -A novel approach for topical delivery. J Pharm Res,2012;5( 3):1755.
Byung-Nak Ahn, Shin-Keun Kim and Chang-Koo Shim. Preparation and evaluation containing Propranolol hydrochloride. J Microencapsul.,1995;12(4):363-375.
Ning MY, Guo YZ, Pan HZ, Yu HM et al., Preparation and evaluvation of proliposomes containing Clotrimazole. Chem Pharm bull (Tokyo), 2005;53(6):620-624.

Wipaporn Rojanarat, Titpawan Nakpheng, Ekawat Thawithong, Niracha Yanyium et al.,Levofloxacin-Proliposomes: Opportunities for use in lung Tuberculosis.Pharmaceutics,2012;4:385-412.
Sandeep Loona, Nitan Bharti Gupta, Khan M V. Preparation and characterization of Metformin proniosomal gel for treatment of diabetes mellitus. Int J Pharm Sci Rev Res,2012;15(2):108-114.
Brock DR, Betageri GV. Enhanced oral absorption of Halofantrine enantiomers after encapsulation in a proliposomal formulation. J Pharm Pharmacol,2002;54(8):1049-1053.
Madhavi M, Maher C P, Pochaiah B, Rao A M .Formulation and evaluation of Metformin based noisome. IJPRR,2013;2(1):1-7.
Chuandi Sun, Ji Wang, Jianping Liu, Wenli zhang. Liquid proliposomes of Nimodipine drug delivery system: Preparation, characterization, and pharmacokinetic.AAPS Pharm Sci Tech,2013;14(1):332-338.
Rajesh Kumar, Ram B, Gupta, Guru V et al., Formulation, characterization and invitro release of glyburide from proliposomal beads. Drug deliv,2001;8(1):25-27.
Manish K Singh, Vikas Chandel, Vandana Gupta and Suman Ramteke.Formulation development and characterization of microemulsion for topical delivery of Glipizide.Der Pharmacia Lettre,2010;2(3):33-42.
Sharma Dinesh Kumar, Gupa vipin Bihari and Purohit Suresh.Invivo evaluation of hyperglycemic activity of solid dispersion of Glibenclamide using pregelatinesed starch as a solubility enhancer. IRJP,Jan 2011;2(1):256-262.
Deepthi Annakula, Madhukar Rao Errabelli, Raju Jukanti, Suresh Bandari et al., Provesicular drug delivery system.Arch Appl Sci Res,2010;2(4):135-146.
Jin-Ming Li, Yan-Zhuo Zhang, Jan-Gang Ren and Yun-Zhi qu. Preparation of stealty Etoposide proliposomes and the pharmakemetics in rabbits. Journal of Chines Pharmaceutical Sciences,2008;17:303-308.

9. / SIGNATURE OF CANDIDATE / (SHRUTHI M V)
10. / REMARKS OF GUIDE / RECOMMENDED AND FORWORDED
11.
/ NAME AND DESIGNATION OF
11.1 Guide
11.2 Signature
11.3 Co guide (if any)
11.4 Signature
11.5 Head of Department
11.6 Signature / Mr.S. PARTHIBAN., M Pharm.,
ASSITANT PROFESSOR,
DEPARTMENT OF PHARMACEUTICS,
BHARATHI COLLEGE OF PHARMACY,
BHARATHINAGARA, MANDYA,
KARNATAKA- 571422.
Not applicable.
Not applicable.

Mr.S.PARTHIBAN.,M.Pharm.,
DEPARTMENT OF PHARMACEUTICS,
BHARATHI COLLEGE OF PHARMACY,
BHARATHI NAGARA, MANDYA,
KARNATAKA-571422.
12. / 12.1 Remarks of the
Chairman and Principal / RECOMMEDED & FORWARDED
12.2 Name and designation of principal
12.3 Signature / Dr.T.TAMIZH MANI.,M.Pharm, Ph.D.
PRINCIPAL,
BHARATHI COLLEGE OF PHARMACY,
BHARATHINAGARA, MANDYA,
KARNATAKA-571422.