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Int. J. LifeSc. Bt & Pharm. Res. 2012 K L Senthilkumar et al., 2012

PREPARATION AND CHARACTERIZATIONOF NABUMETONE LIPOSOMES

K L Senthilkumar1*, R P Ezhilmuthu2, Praveen P2

Research Paper

Nabumetone is the effective drug for the treatment of reumatoid arthritis and osteoarthritis. Thedrug has a half life of 23 hrs and its oral bioavailability is only 30%. The aim of the study is toprepare 6 formulations of liposomal carrier for Nabumetone for the treatment of arthritis that iscapable of delivering the drug to the specific target site by topical route by using different ratiosof phospho lipid and cholesterol with a desired amount of drug by thin film hydration techniqueand to find out the drug release from the liposome's of different ratios, mechanism kinetics ofdrug release pattern and also to find out the size distribution of liposome's of different ratios andalso to increase the bioavailability and efficacy of the drug.

Keywords: Liposome's, Thin Film Hydration Method, Nabumetone.

*Corresponding Author: K L Senthilkumar,[email protected]

INTRODUCTIONLiposomes are colloidal particles formed as

concentric bimolecular layers that are capable of

encapsulating drugs. They are lipid vesicles that

fully enclose an aqueous volume. These lipid

molecules are usually phospholipids with or

without some additives. Cholesterol may be

included to improve bilayer characteristics of

Liposome’s; increasing micro viscosity of the

bilayer, reducing permeability of the membrane

to water soluble molecules, stabilizing the

membrane and increasing rigidity of the vesicle.

Liposomes are micro vesicles, whose

membranes are made of phospholipids, which

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Int. J. LifeSc. Bt & Pharm. Res. 2012

1 Department of Pharmaceutics, Padmavathi college of pharmacy and Research institute, Dharmapuri.2 Department of Pharmaceutics, Padmavathi college of pharmacy and Research Institute, Dharmapuri.

are bilayer having head and tail. The head group

is hydrophilic and the tail group which is made of

long hydro carbon chain is hydrophobic. The drug

molecules can be encapsulated in aqueous

space or into lipid bilayer. The exact location of

the drug will depend upon its physicochemical

properties and composition of lipids. The water

soluble substances can be trapped inside the

water sphere while fat soluble substances can

be trapped inside the fat soluble opposite end of

the molecule. Liposome’s give a unique delivery

system for nutrients, vaccines, enzymes, or

drugs. Liposomes are effective in treating

diseases that affect the phagocytes of the immune

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system because they tend to accumulate in the

phagocytes, which recognize them as foreign

invaders. They have also been used to carry

normal genes into a cell in order to replace

defective, disease-causing genes. Because of

their moisturizing qualities Liposome’s are also

used in cosmetics.

Liposome’s, which are vesicles consisting of

one or more concentrically ordered assemblies

of phospholipids bilayer, range in size from a

nanometer to several micrometers. Phospholipids

such as egg phosphatidylcholine, phosphatidyl-

serine, synthetic dipalmitoyl-DL-alpha-

phosphatidylcholine or phosphatidylinositol, have

been used in conjunction with cholesterol and

positively or negatively charged amphiphiles such

as stearylamine or phosphatidic acid. Because

of the multifold characteristics as drug carriers,

Liposome’s have been investigated extensively

as carriers of anticancer agents for the past

several years. Liposomal entrapments include a

variety of pharmacologically active compounds

such as antimalarial, antiviral, anti-inflammatory

and anti-fungal agents as well as antibiotics,

prostaglandins, steroids and bronchodilators.

MATERIALS AND METHODSNabumetone drug, Soya lecithin 30% Cholesterol,

Chloroform, Methanol Potassium dihydrogen

phosphate, Disodium hydrogen phosphate,

Sodium chloride, Acetone were used for this study.

Methods

Formulation of Liposomes

Drug: Soya lecithin: Cholesterol in different ratios

was dissolved in 5 ml chloroform. This solution

was taken in a 250 ml round bottom flask. The

flask was rotated in rotary flash evaporator at 80

rpm for 15 minutes in thermostatically controlled

water bath at 40°C under vacuum 900 mmHg.

The organic solvent was slowly removed by this

process such that a very thin film of dry lipids

was formed on the inner surface of the flask. The

dry lipid film was slowly hydrated with 5 ml of

phosphate buffer ph 7.4.the fluid is allow to

hydrate with phosphate buffer pH 7.4 for 2 hr

Swirling the contents to yield milky white

suspension. The formulation is subjected to

centrifugation. The unentrapped drug is removed

by centrifugation at 3000 rpm for a period of 30

minutes. The pellets are dispersed in phosphate

buffer.

CHARACTERIZATION OFLIPOSOMESDrug Entrapment Efficiency

Drug entrapment efficiency was calculated by

using centrifugation method. The liposome

suspension of 1ml was taken and centrifuged at

3500rpm for 15 min. The sediment obtained from

the centrifugation was suspended in 100 ml of

phosphate buffer pH 7.4. Then the absorbance

was taken at 261 nm. From that the amount

present in 1 ml of suspension was obtained. The

drug entrapment efficiency was calculated from

the following formula.

Amount of Drug in the sediment% Drug Entrapped (PDE) = ×100

Total amount of Drug

Particle Size Analysis

The particle size of Liposome’s was

determined by optical microscopy by using Perkin

Elmer. All the prepared batches of Liposome’s

were viewed under microscope to study their size.

Size of liposomal vesicles was measured at

different location on slide by taking a small drop

of liposomal dispersion on it and average size of

liposomal vesicles were determined

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In Vitro Drug Release Study

The release studies were carried out in diffusioncell having 10 ml capacity. 10 ml Phosphate bufferpH 7.4 was placed in diffusion cell. The diffusioncell contained a magnetic bed and the mediumwas equilibrated at 37±50C. Dialysis membranewas taken and placed on the diffusion cell. Afterseparation of non-entrapped Nabumetone,liposome dispersion was filled in the dialysismembrane. The dialysis membrane containingthe sample was suspended in the medium.Aliquots were withdrawn (1 ml) at specificintervals, f iltered, diluted with phosphatebuffer and the absorbance was taken at 261 nm.Then the apparatus was immediately replenishedwith same quantity of fresh phosphate buffer pH

7.4 medium.

RESULTS AND DISCUSSIONFTIR of pure drug and physical mixture.

The drug release kinetics of prepared

Nabumetone Liposomes were conducted using

Higuch's model and Poppa's model. Higuch's plot

was done with square root of time in X axis and

%cumulative drug in Y axis (Figure 1). The

Poppa's plot was done with log time in X axis and

log %cumulative drug release in Y axis. The study

of drug release kinetics showed that majority of

the formulations governed by peppas model.

Results showed that F1 have a particle size

of 7.994 micron and F6 have a highest value com-

pared to other formulations. From the results of

particle size analysis F1 shows satisfactory

particle size (Figure 2).

According to the drug entrapment study

conducted the maximum drug entrapment was

shown by F1 (52±0.5).

Figure 1: FTIR of Pure Drug and Physical Mixture

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Figure 2: In Vitro Drug Release Study of F1 to F6

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Figure 3: SEM Photographs of Liposome’s

CONCLUSIONNabumetone Liposome's were prepared using

soya lecithin, cholesterol and chloroform as

solvent by thin film hydration method using rotary

evaporator. The prepared Liposomes were

evaluated by drug entrapment study, particle size

analysis. In vitro drug release study and

mechanism of release kinetics using Higuchi's

plot and Korsemeyer Peppas plot.

The present study demonstrated the

successful preparation of Nabumetone Liposomes

and its evaluation. Formulation F1 showed high

encapsulation efficiency with minimum particle

size and drug release over a 4 hr, hence suppose

to give greater bioavailability and considered as

good liposomal formulation. The difference in

drug entrapment and drug release was due to

the different ratios of lipid to cholesterol. The drug

entrapment shows decreasing when the

concentration of lipid and cholesterol changes.F1

have high entrapment and satisfactory drug

release within 4 hr. F1 shows least particle size

from the results F1 selected as a good formulation

of Nabumetone Liposomes.

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