Formulation and Evaluation of Gliclazide Loaded Ethosome As Transdermal Drug Delivery Carriers

FORMULATION AND EVALUATION OF GLICLAZIDE LOADED ETHOSOME AS TRANSDERMAL DRUG DELIVERY CARRIERS

Synopsis for M.Pharm Dissertation submitted to the

Rajiv Gandhi University of Health Sciences Karnataka, Bangalore.

By

Mr. VIJAYA KUMAR K S

M.Pharm., Part-I

Under the guidance of

Mr. S. PARTHIBAN., M. Pharm.,

Department of Pharmaceutics,

BharathiCollege of Pharmacy,

Bharathinagara.

2013-2014

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA,BANGALORE.

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. / NAME OF THE CANDIDATE
AND ADDRESS / VIJAYAKUMAR K S 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 EVALUATION OF GLICLAZIDE LOADED ETHOSOMES AS TRANSDERMAL DRUG DELIVERY CARRIERS
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 / Brief resume of the intended work:
ENCLOSURE – I
6.1 – NEED FOR THE STUDY
Ethosomes are noninvasive delivery carriers that enable drugs to reach the deep skin layers and/or the systemic circulation. These are soft, malleable vesicles tailored for enhanced delivery of active agents. They are composed mainly of phospholipids, (phosphatidylcholine, phosphotidylserine, phosphatitidic acid), high concentration of ethanol and water. The high concentration of ethanol makes the ethosomes unique, as ethanol is known for its disturbance of skin lipid bilayer organization; therefore, when integrated into a vesicle membrane, it gives that vesicle the ability to penetrate the stratum corneum. Also, because of their high ethanol concentration, the lipid membrane is packed less tightly than conventional vesicles but has equivalent stability, allowing a more malleable structure and improves drug distribution ability in stratum corneum lipids.1,2
Transdermal drug delivery system is more pertinent in case of chronic disorders such as hypertension and diabetes, which require long term dosing to maintain therapeutic concentrations. Several studies have been reported for transdermal drug delivery of anti diabetic drug to enhance the bioavailability as well as to improve the patient compliance. Transdermal drug delivery (TDD) is gaining prominence over other forms of drug delivery due to its potential advantages, including reduced systemic side effects, noninvasiveness, increased patient compliance, large area of interface, potential for continuous and controlled delivery. Consequently, in recent years, numerous transdermal products have been introduced into the market. Annual sales worldwide are estimated to be $31.5 billion by 2015. Still, only few drugs are presently available with transdermal patches fundamental reason why so few drugs are used is that the barrier property of the skin limits the use of patches to therapeutics, where the molecule size is small enough to diffuse through the skin at therapeutic rates and the other important issues is that the use of penetration enhancers to increase the flux of the drug.3
Hence there is a need of alternative novel drug delivery system which improve the drug penetration of drug through skin barrier is essential. Different reports shows a promising future of ethosomes in making transdermal delivery of various agents more effective to penetrate the skin efficiently. Ethosomes also offer a good opportunity for the non-invasive delivery of small, medium, and large sized drug molecules. Preparation of ethosomes is easy with no complicated equipment involved and therefore can be scaled up to the industrial level. 4,5,6
The prevalence of type 2 diabetes is about 12% in urban India, and it is estimated that the country has the largest number of diabetic patients in the world. Currently available antidiabetic agents have relatively short half-life, low bioavailability and poor retention, and undesirable side effects. It is a challenge for people with diabetes to keep track of their blood sugar levels and to maintain right amount of insulin throughout the day. However, the basal rate of insulin needed by diabetics is 0.5–1 mg/day. These drawbacks give researchers tremendous opportunities to design and develop novel drug delivery systems to overcome the transport barriers, inherent elimination and metabolism problems associated with these antidiabetic drugs. Therefore, the search for more effective and safer hypoglycemic agents has continued to be an important area of active research7,8,9
Transdermal systems are ideally suited for diseases that demand chronic treatment. Hence, anti-diabetic agents of both therapeutic and prophylactic usage have been subjected to transdermal investigation. Gliclazide is a second-generation sulfonyl urea oral hypoglycemic agent used in the treatment of non-insulin-dependent diabetes mellitus. But the problem with this potentially useful hypoglycemic agent is that it is practically insoluble in water. This limits its oral bioavailability with large individual variation. After absorption it gets extensively metabolized by hydroxylation, N-oxidation, and oxidized to several inactive metabolites. It is slightly soluble in water having half-life 6-8 hrs. The drug is neutral in nature, molecular weight 323.4, melting point about 181ºC, and partition coefficient 2.1. As the drug contains ideal properties required for transdermal preparation, it can be used for present work.10,11.
The aim of the study is to develop novel drug delivery ethosomes of Gliclazide and evaluate ethosomal drug delivery system of Gliclazide for o vercome the problem associated with oral delivery of Gliclazide with enhanced antidiabetic activity.
ENCLOSURE - II
REVIEW OF LITERATURE
Rathore A R et al., investigated Repaglinide entrapped ethosome and dispersions were produced by cold and hot method. They reported that the morphology and dimensional distribution of the disperse phase have been characterized by scanning electron microscopy and photon correlation spectroscopy, respectively and an in-vitro diffusion study was conducted by Franz cell associated to stratum corneum epidermis membrane on ethosome dispersions viscosized by Carbopol 934. They studied Effect of different concentration of lipid and ethanol on different properties of ethosomal formulations and concluded that ethosomes were a promising candidate for transdermal delivery of Repaglinide being a safe and very efficient drug carrier for transdermal delivery of drug.12
Navneet bhulli et al., investigated entrapped Glimepiride in novel vesicular carrier system (Ethosomes) to improve therapeutic efficacy of Glimepiride via transdermal route. They characterized and evaluated the transdermal potential of ethosomes by Vesicular shape, surface morphology and entrapment efficiency were determined by optical microscopy, transmission electron microscopy (TEM), and minicolumn centrifugation technique respectively. Their results shown that The ethosomal formulation were found to be more efficient delivery carriers with high entrapment and optimal nanometric size range and low polydispersity index in comparison with plain drug solution and liposomal formulation. 13
Bodade SS et al., prepared ethosomes as mode for transdermal delivery of Repaglinide (RPG) by cold method using dipalmitoyl phosphatidylcholine and ethanol. They have reported that the ethosomal system can successfully deliver RPG transdermally, sustain its effect and thus reduce its dosing frequency and also enhancing the efficacy of RPG in the treatment of diabetes.14
Suman Ramteke et al,.Glipizide based micro emulsion was developed and its usefulness as topical drug carrier system for the non-insulin dependent diabetes mellitus (NIDDM).Inclusion complex showed significant blood Glucose reduction as compared to drug alone. Micro emulsion system provides viscous consistency for the topical application, which delivered the drug in sustained or controlled manner and prolonged delivery as compared to conventional dosage form.15
Suresh c. Joshi et al., discussed valuable information regarding strategies used in transdermal drug delivery systems and antidiabetic approach of this novel drug delivery system and provide an overview on various techniques and new anti–diabetic approach.16
 Atul Kumar Garg et al., studied the effect of different concentrations of Lecithin and ethanol on different properties of Aceclofenac incorporated ethosomes .They reported that the size of the vesicles was found to have increased with increasing lecithin concentration and the size of the vesicles decreased significantly with increasing ethanol concentration. They reported ethosomes with 3% lecithin and 20% ethanol were shown highest Aceclofenac release (92.72 ± 1.04%).17
 Raj Kumar Tiwari et al., has reviewed ethosomes are soft, malleable vesicles tailored for enhanced delivery of active agents. Ethosomes provides a number of important benefits including improving the drug's efficacy, enhancing patientcompliance and comfort and reducing the total cost of treatment.18
 Kumar R et al., have reviewed several methods to increase the permeation rate of drug temporarily and suggested one simple and convenient approach is application of drugs in formulation with elastic vesicles or skin enhancers.19
 Donatella Paolino et al.,studied the evaluation of various ethosomal suspensions made up of water, phospholipids and ethanol at various concentrations for their potential application in dermal administration of Ammonium Glycyrrhizinate, a useful drug for the treatment of various inflammatory-based skin diseases.20
 Barupal A Ket al.,developed ethosomes of Aceclofenac by varying the quantity of ethanol 10-50% (v/v),lecithin 1-4% (w/v), propylene glycol 5-20% (v/v).They reported ethosomes of average size of 1.112 μm with a spherical shape bearing smooth surface which were observed by transmission electron microscopy and surface electron microscopy.21
 Jin-Guan Chen et al., studied the filming-rehydration and ultrasonic method combined to prepare ethosomes as a carrier of Triptolide.22
 Upadhyay N et al., reviewed that the ethosomes have higher penetration rate through the skin as compared to liposomes and it can be used widely in place of liposomes. Ethanol increases the cell membrane lipid fluidity which results in increased skin penetrability of the ethosomes. Ethosomes are able to encapsulate and deliver through the skin highly lipophilic molecules such as cannabinoids, Testosterone and Minoxidil, as well as Propanolol, Trihexyphenidil, Cyclosporine A, Insulin, Salbutamol, etc.23
Vijay Kumar M.R., et al., successfully formulated and evaluated Diclofenac potassium ethosomes, and reported the three dimensional nature of ethosomes, which has the smaller vesicular size than the liposomes prepared without alcohol.24
 Ashoniya Sheer et al., prepared Ketoconazole entrapped ethosomal carriers. They have studied the effect of different concentration of lecithin and ethanol on different proportions.25
 Rahul G.S. Maheshwari et al., formulated and evaluated the transdermal potential of novel vesicular nanocarriers. Ethosomes and ultra-deformable liposomes, containing Clotrimazole, an anti-fungal bioactive. They reported ethosomal formulation and ultradeformable liposomal formulation showed highest entrapment, optimal nanometric size range and smallest polydispersity index.26
 Sui Yung Chan et al., prepared, optimized and characterized the potential of ethosomes for delivery of Ketoprofen via skin. Their results demonstrated that the ethosomal carrier significantly improved the delivery due to 5-aminolevulinic acid and the formation of Protoprsophyrin IX in both normal and hyper proliferative murine skin samples, and the expression level of tumor necrosis factor (TNF)-α was reduced after the containing 5-aminolevulinic acid –ethosomes were applied to treat hyper proliferative murine skin.27
 Touitou Tet al., investigated that, the ethosomal system, which is composed of phospholipids, ethanol and water. They reported ethosomal systems were much more efficient at delivering a fluorescent probe to the skin in terms of quantity and depth, then their liposomes or hydro alcoholic solution .and dramatically enhanced the skin permeation of mice.28
 Michael H. Quist.et al., have studied effect of chemical permeation enhancers from drug-in-adhesive transdermal patches. In this study, the release of eight commonly known enhancers from eight types of polymer adhesives was evaluated using Franz diffusion cells. It was reported that all the enhancers released completely from the adhesives and followed a square root of time kinetic (Higuchi law).29
 Peeush Singh et al,.have reviewed on the novel technique to enhance therapeutic efficacy and safety of drugs in transdermal drug delivery system. They suggested that the conventional oral dosage forms has significant drawbacks of low bioavailability due to hepatic first pass metabolism and tendency to produce rapid blood level spikes (Both high and low), leading to a need for frequent dosing, which can be both cost ineffective and inconvenient and to improve such characters transdermal drug delivery system (TDDS) had been emerged.30
Anilkumar J. Shinde et al,. Studied the effect of matrix type patches prepared by film casting technique of Gliclazide. Their release study revealed that the patch containing inclusion complex of the drug with hydroxypropyl β-cyclodextrin, eudragit-RL 100, hydroxypropyl methyl cellulose (HPMC), and chitoson reported higher permeation compared with hydroxypropyl β-cyclodextrin, eudragit-RL 100, HPMC and chitosan. 31
 Deepak Shrivastava et al,.reviewed that ransdermal drug delivery system (TDDS) provides a means to sustain drug release as well as reduce the intensity of action and thus reduce the side effects associated with its oral therapy. Transdermal drugs are self-contained, discrete dosage form. Diabetes mellitus is a chronic metabolic disorder characterised by high blood glucose concentration hyperglycaemia caused by insulin deficiency.To optimize this drug delivery system, greater understanding of the different mechanisms of biological interactions, and polymer are required. TDDS a realistic practical application as the next generation of drug delivery system.32
 Sujatha S et al., investigated antidiabetic effect of methanolic extract of flowers of antigen leptopus (MFAL) and evaluated at three doses i.e.100, 200 and 400 mg/kg , p.o. in normal, glucose fed and alloxan-induced diabetic rats. They compared hypoglycemic and anti-diabetic effect of the MFAL with Glibenclamide, a reference drug used in type 2diabetes therapy. 33
Sharma Dinesh Kumar et al., prepared solid dispersed Glibenclamide using pregelatinesed starch by physical and kneading method are reported that shows enhanced dissolution and hypoglycemic activity.34
Ravi Teja Allena et al., developed a sustained release transdermal patch of Metformin hydrochloride using a natural polymer like chitosan and a hydrophilic polymer like HPMC and reported that this system can be efficient for the drugs which have the similar properties as that of Metformin HCl i.e. which causes gastric irritation and having high first pass metabolism, and freely water soluble in nature .From this study they suggested that applying transdermal patches containing Metformin HCl achieved sustained drug delivery without gastric irritation.35
ENCOSURE III
6.3 OBJECTIVE OF STUDY:
The specific objective of the study is to formulate and evaluate the ethosomal gel of Gliclazide for transdermal drug delivery.

• To formulate ethosome entrapped with Gliclazide 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 ethosomal gel using Carbopol gel base.
• To evaluate the prepared ethosomalgelfor 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.

7.0 / MATERIALS AND METHODS:
DRUG: Gliclazide.
POLYMER: Carbopol (or) any other suitable polymer will be selected.
CHEMICALS: Soya phosphatidyl choline, cholesterol, ethanol, chloroform, disodium hydrogen phosphate, potassium dihydrogen phosphate, triton -100, polyvinyl alcohol, polyvinyl pyrolidon and tri ethanolamine, etc
METHOD :

1. Preparation of ethosomes by cold method/ hot method, any other suitable methods

and incorporation of optimized ethosomal vesicles in gel
Evaluation:
Compatibility study by Fourier Transform Infra Red (FTIR)/ Differential Scanning Calorimetry (DSC).
Surface morphology by scanning electron microscope (SEM)/ (TEM).
Vesicular size and size distribution.
Drug entrapment efficiency.
Zeta potential determination.
Content uniformity of gel.
In-vitro diffusion studies using Franz diffusion cell.
In-vitro release kinetics modeling.
Stability study.
Perform skin irritation test
ENCLOSURE - IV
7.1 Source of Data:

1. Research publications.
2. International and Indian Journals.
3. Textbooks and reference books.
4. RGUHS Library.
5. Library: Bharathi College of Pharmacy.

a. Journals
Asian Journal for Control Release.
Biophysical Journal.
Journal for Control Release.
Journal of Herbal Medicinal and Toxicology.
Asian Journal of Pharmaceutical Science and Technology.
International Journal of Pharmaceutical Science and Research.
International Journal of Pharmaceutical and Biological Archives.
International Journal of Therapeutic Application.
Der Pharmacia Lettre.
International Journal of Drug Delivery and Research.
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ENCLOSURE - V
7.2 Method of collection of data:
1.The data for the study is planned to collect from the laboratory-based experiments
2.Preformulation studies like solubility, melting point and characterization of the drug and phospholipids will be done by employing suitable methods.Compatibility of drug with phospholipids will be carried out by using Infra-Red Spectroscopy and Differential Scanning Calorimetry instruments adopting reported methods.

1. Preparation of Gliclazide loaded ethosomes by cold/hot method or any other suitable Methods.
2. Methods for the characterization of ethosomal formulation :

(a) Vesicle shape (morphology)
Transmission electron microscopy or scanning electron microscopy or any other suitable methods.
(b) Entrapment efficiency
Mini column centrifugation method or fluorescence spectrophotometer or any other suitable methods.
(c) Vesicle size and size distribution
Optical microscopy method or any other suitable methods
(d) Zeta potential
Zeta meter or any other suitable methods.
(e) In Vitro drug release study
Franz diffusion cell with artificial or biological membrane, or dialysis bag diffusion method or any other suitable methods.