FORMULATION AND EVALUATION OF SUSTAINED RELEASE MICROSPHERES

OF FENOFIBRATE

Done by

Mini Mol.P.V

Reshma Fathima.K

Under the Guidance of

Ms.Sreethu K Sreedharan

CONTENTS

Introduction

Literature Review

Objective And Plan of Study

Materials And Equipments

Methodology

Results And Discussion

Summary And Conclusion

References

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INTRODUCTION

A well designed controlled drug delivery system can overcome some of the problems of conventional therapy and enhance the therapeutic efficacy of a given drug. To obtain maximum therapeutic efficacy, it becomes necessary to deliver the agent to the target tissue in the optimal amount in the right period of time there by causing little toxicity and minimal side effects.

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There are various approaches in delivering a therapeutic substance to the target site in a sustained controlled release fashion.

One such approach is using microspheres as carriers for drugs.

Microspheres are characteristically free flowing powders consisting of proteins or synthetic polymers which are biodegradable in nature and ideally having a particle size less than 200 μm.

This is the important approach in delivering therapeutic substance to the target site in sustained and controlled release fashion.

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Microspheres are sometimes referred to as microparticles. Microspheres can be manufactured from various natural and synthetic materials.

Polyethylene, polystyrene and expandable microspheres are the most common types of polymer microspheres.

Polystyrene microspheres are typically used in biomedical applications due to their ability to facilitate procedures such as cell soeting and immunio precipitation

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Mechanism of Drug Release

Most of the drug delivery through micro particles inhibits a matrix type internal solid dispersion morphology structure.

The drug may be insoluble in the polymeric matrix and the drugs are released by erosion. Initially water diffuses into the matrix dissolving the resulting adjacent to the surface of the device.

The resulting osmotic pressure is relieved by forming a channel to the surface releasing a defined amount of drug in the initial drug burst.

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Materials Used

Microspheres used usually are polymers. They are classified into two types

1. Synthetic polymers

2. Natural polymers

Synthetic Polymers:

a)Non biodegradable polymers:

Polymethyl methacrylate (PMMA), Acrolein, Glycidyl methacrylate, Epoxypolymers

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b) Biodegradable polymers:

Lactides, their glycolides and their copolymers, PolyalkylCyano Acrylate.

Natural polymers

These are obtained from different sources like proteins, carbohydrates and chemically modified carbohydrates.

Proteins: Albumin, Gelatin, And Collagen, Carbohydrates: Agarose, Carrageenan, Chitosan, Starch.

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Methods

Single emulsion technique

Double emulsion technique

Polymerization

Phase separation/ Coacervation

Spray drying

Solvent extraction

Emulsion Solvent Evaporation

Wet emulsion technique

Hot Melt Microencapsulation

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Pharmaceutical Applications

Ophthalmic Drug Delivery Polymer Gene delivery Gene delivery systems Intratumoral and local drug delivery Oral drug delivery Nasal drug delivery Buccal drug delivery Gastrointestinal drug delivery: Peroral drug delivery Vaginal drug delivery Transdermal drug delivery Colonic drug delivery Multiparticulate delivery system

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LITERATURE REVIEW

Naeem. M, Kiran. B et al., 2013 developed the fenofibrate loaded liposphere system. An attempt was made to improve aqueous solubility of FNO by aid of stearic acid and Paraffin oil. Significant improvement in the aqueous solubility of the drug in the FNO lipospheres supports the applicability of lipospheres as a tool for improving aqueous solubility of the BCS class-II drugs10.

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Modi, Tayade et al 2006., developed the Solid dispersion (kneading) technique for the enhancement of the dissolution profile of valdecoxib using solid dispersion with PVP and the preparation of fast-dissolving tab- lets of valdecoxib by using a high amount of superdisin- tegrants. A phase solubility method was used to evaluate the effect of various water-soluble polymers on aqueous solu-bility of valdecoxib. Polyvinyl pyrrolidone (PVP K-30) was selected and solid dispersions were prepared by the method of kneading14.

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Drug Profile

DRUG : Fenofibrate

CATEGORY : Hypolipidaemic drug

MOLECULAR STRUCTURE :

MOLECULAR FORMULA : C20 H21ClO4

DESCRIPTION: white crystalline solid, odourless.

MELTING POINT: 79-82°C

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SOLUBILITY: Soluble in organic solvents such as ethanol, methanol, DMSO, and DMF. Sparingly soluble in aqueous buffers.

MECHANISM OF ACTION: it is a fibrate class hypolipidaemic drug. It enhances activity of the enzyme lipoprotein lipase, which degrades VLDL level resulting in lowering of triglycerides. They also increase HDL levels. Fibrates also inhibits coagulation and promote thrombolysis, which also accounts for their beneficial effects.

ADVERSE EFFECTS: GI upsets, skin rashes, headache, muscle cramps blurred vision.Adverse events occurs in about 1% of the patients treated with fenofibrate. Skin rashes were the most frequent event causing discontinuation of therapy.

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PHARMACOKINETICS: pharmacokinetics significantly affected by food intake. The high fat breakfast will significantly affecting the rate of absorption of fenofibrate more than the standard breakfast and fasted conditions specifically AUC from zero to infinity and peak plasma concentration (Cmax) is increased.

DOSE: Orally available as 140mg, 160 mg and 200mg tablets and capsules.

STORAGE: Store below 30ºC.

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OBJECTIVE AND PLAN OF STUDY

The sustained release dosage forms are designed to achieve a prolonged therapeutic effect by continuously releasing medication over extended periods of time after administration of a single dose.

The ideal drug delivery system will possess two main properties

»It will be a single dose for the whole duration of treatment.

»It will deliver the active drug delivery at the site of action.

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MATERIALS AND EQUIPMENTS USED

SL.N

OMATERIALS SUPPLIED BY

1 FenofibrateAlkem pharmaceuticals Pvt

Ltd,mumbai

2 Gelatin Nice chemicals,Banglore

3 Methanol Nice chemicals,Banglore

4 Tween 20 Prowess lab chemicals

5 Liquid paraffin Prowess lab chemicals

6 Formaldehyde Nice chemicals,Banglore

7 Hydrochloric acid Nice chemicals,Banglore

8 Monosodium phosphate Nice chemicals,Banglore

9 Disodium phosphate Nice chemicals,Banglore

SL.N

OINSTRUMENTS MANUFACTURER

1 Electronic weighing balance Shimadzu BL220H,Japan

2 Magnetic stirrer Remiequipments Pvt Ltd

3 Optical microscope Dolar US 4

4 UV-Spectroscopy Shimadzu 1600,Japan

5 FTIR Shimadzu 8400,Japan

6 Dissolution test apparatus Electro Lab,TDT 08L (USP)

7Differential Scanning

Calorimetry

Shimadzu D.S.C TA60 WS

Thermal Analyzer

8 Scanning Electron Microscopy DSC-823e, Mettler Toledo

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Preformulation Studies

Identification of drug

FTIR (fourier transforms infrared spectra)

DSC Analysis

Physical Appearance

Melting point

Drug excepient compatability studies

Spectrophotometric Scanning of fenofibrate and polymer

Preparation of standard stock solution

Construction of calibration curve

METHODOLOGY18

• Formulation

Preparation of Drug Polymer phase

Preparation of Oily Phase

Preparation of Gelatin microspheres of fenofibrate

Formulation Drug(mg) Polymer(mg)Tween20(

ml)

F1 200 1000 1

F2 200 1500 1

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• Evaluation Studies

Bulk characterization

angle of repose

θ = tan−1 h r

bulk density

Bulk Density = weight of sample in gm

volume occupied by the sample

tapped density

Tapped Density = weight of sample in gm

volume occupied by the sample after tapping

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Carr´s Index (CI)

Carr’s Index = tapped density−bulk density

tapped density

Percentage yield (%Y)

% Yield = practical yield

theoretical yield×100

Percentage Drug Entrapment(%DE)

% Drug Entrapment = practical drug content

theoretical drug content× 100

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Morphology

By using SEM analysis

Invitro Cumulative Percentage Drug Release Study

The FNO release from the microspheres was evaluated by using the US Pharmacopoeia Dissolution Apparatus-II Paddle (XVIII) in 900ml mixture of PBS pH 6.8 with 1% SLS in 9:1 at 37°C ± 0.5 temperature. The rotational speed of dissolution apparatus was maintained at 100rpm. Each run was carried out in triplicates.

Accurately weighed 243mg of microspheres were filled in a “0” size capsule to get the final weight of 475mg each of the capsules was transformed into dissolution media. The 5ml samples were withdrawn at predetermined time intervals with dissolution media replacement and were filtered through 0.45µm whatman filter paper. The drug content was determined spectrophotometrically at 290nm on shimadzu UV/ Vis- spectrophotometer.

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RESULTS AND DISCUSSION

IR spectrum of Fenofibrate

IR spectrum of Gelatin

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IR spectrum of Fenofibrate + Gelatin

Frequency (cm-1)

FNO

Frequency (cm-1)

Physical mixtureInterpretation

3036 3039 Alkanes

3220 3211Monosubsituted benzene

ring

3437 3439 Phenols

3526 3514 1,4-Disustituted phenol

1416 1417 -CH3 group

1622 1625 Carboxylic group

1796,1224 1796,1225 Aromatic ester

1220 1225 -CH2Cl

IR Spectral peaks of Fenofibrate and physical mixture

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SEM Analysis of fenofibrate and microspheres

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DSC analysis of prepared microspheres

DSC Curve of Fenofibrate

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Standard curve

CONCENTRATION ABSORBANCE

0 0.00

10 0.553

20 0.996

30 1.545

40 2.015

50 2.497

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Bulk characterization

Sl.no Parameter

Fenofibrate microspheres

F1 F2

1 Angle of repose 24.11 25.22

2 Bulk density 0.26 0.31

3 Tapped density 0.33 0.39

4 Carr’s index 20 22

5 Hausner’s ratio 1.25 1.30

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Percentage yieldThe percentage yield was calculated as per the equation and it was found to be

74 % and 76% in formulation F1 and F2 respectively.

Percentage Drug Entrapment% Drug entrapment of drug entrapped within the polymer matrices were in the range of 70-97 %. An entrapment efficiency depends on the drug solubility in the solvent system used for processing.

In-vitro drug release study: The drug release from microspheres in phosphate buffer pH 6.8 has been shown in Figure . Cumulative % drug release from F1and F2, were in the range of 60 and 68% within 12hours respectively. No formulation is showing burst release which indicates the absence of free particles on the surface of microspheres which further confirmed by SEM study. The trail revealed that low level of gelatin (25%) failed to produce microsphere with acceptable physical characteristic where as high level of stearic acid (100%) resulted in liposphere that exhibited high percentage of drug release. As the level of paraffin oil increases the particle size of liposphere increases due to tackiness which in turn decrease the % drug release from the liposphere

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Dissolution values of F1 Dissolution Values of F2

Sl.n

o

Time

(hr)

Absorbanc

e

Concentrati

on (µg/ml)

Amount of

drug

released

% drug

release

Cumulativ

e % drug

release

1 0 0 0 0 0 0

2 0.5 0.009 0.0611 0.0611 4.07 4.07

3 1 0.020 0.1919 0.1919 12.79 12.87

4 2 0.038 0.4058 0.4058 27.05 27.31

5 3 0.045 0.4890 0.4890 32.60 33.14

6 4 0.052 0.5722 0.5722 38.15 38.79

7 5 0.060 0.6673 0.6673 44.49 45.25

8 6 0.067 0.7506 0.7506 50.04 50.93

9 7 0.072 0.8099 0.8099 53.99 54.99

10 8 0.073 0.8219 0.8219 54.79 55.87

11 10 0.075 0.8456 0.8456 56.37 57.47

12 12 0.079 0.8932 0.8932 59.15 60.17

Sl.n

o

Time

(hr)

Absorban

ce

Concentrati

on (µg/ml)

Amount of

drug

released

%

drug

releas

e

Cumulati

ve % drug

release

1 0 0 0 0 0 0

2 0.5 0.009 0.0574 0.0574 5.45 5.45

3 1 0.013 0.1068 0.1068 7.25 7.29

4 2 0.019 0.1962 0.1962 14.41 14.49

5 3 0.028 0.2869 0.2869 19.13 19.34

6 4 0.035 0.3702 0.3702 24.68 25.06

7 5 0.050 0.5485 0.5485 36.57 37.06

8 6 0.062 0.6911 0.6911 46.07 46.80

9 7 0.074 0.8338 0.8338 55.59 56.51

10 8 0.081 0.9169 0.9169 61.13 62.24

11 10 0.086 0.9764 0.9764 64.09 65.26

12 12 0.090 1.0239 1.0239 66.26 67.39

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SUMMARY AND CONCLUSION

Following concclusions have been drawn from the present study:

In the preformulation studies, it was found that Fenofibrate is having poor flow property. Hence in the present study, the resulting microsphere considerably improved flow properties.

The analytical mehod used in the present study was found to be suitable for the estimation of Fenofirate, which was indicated by the high regression values obtained in the standard plot.

The yellowish white microspheres were obtained by using emulsion – coacervationmethod and the percentage yield was found to be 74% for F1 and 76 % for F2.

Scanning electron microscopy of Fenofibrate microsphere was also performed and the surface morphology were studied.

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Sustained release property of microsphere were also enhanced by incorporating the microsphere in enteric coated capsules.

The percentage drug entrapment were in the range of 70-97%.

The invitro drug release study showed that the microspheres were released 60 and 67% of drug at 12 hrsin formulation F1 and F2 respectively.

Gelatin can be effectively used for the preparation of sustained release Fenofibrate microsphere. In conclusion, the present study demonstrated the successful preparation of once daily sustained release microsphere of Fenofibrate.

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REFERENCES

Ramteke. K. H, Jadhav.V. B, Dhole. S. N, [2012], Microspheres as carriers used for novel drug delivery system, IOSRPHR, 2(4), 44-48.

Sahil, Akanksha, Premjeet, Bilandi, Kapoor, [2011], Microsphere a review, IJRPC, 1(4).

Prabus. L, Shirwaikar A, Kumar, [2009], Formulation and evaluation of sustained release microspheres of rosin containing aceclofenac, ARS Pharm, vol 50, 51-62.

Gangadhar.C. B, Sunder. R, Varma. M, Raju. M, Saikiran. M, [2010], Formulation and evaluation of indomethacin microspheres using natural and synthetic polymers as controlled release dosage forms, International journal of drug discovery, 2(1), 8-16.

Barhate. D, Rupnar. S, Sonvane. M, Pawar. R, Rahane. D, [2009], Formulation and evaluation of floating microspheres of ketorolac trometSSSamol, International journal of pharmaceutical research and development online,1 (9), ISSN: 0974-9446.

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THANKS

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