EFFERVESCENT PROLIPOSOMES FOR AEROSOL DELIVERY TO PARANASAL SINUSES BY OSHADIE KORALE A thesis submitted in partial fulfilment for the requirements for the degree of PhD at the University of Central Lancashire June, 2016 Dedication I dedicate this achievement to my parents, fiancé, brother, my grandmother and friends; without their encouragement and love this would not have been possible. 1 University of Central Lancashire STUDENT DECLARATION FORM Concurrent registration for two or more academic awards I declare that while registered as a candidate for the research degree, I have not been a registered candidate or enrolled student for another award of the University or other academic or professional institution ____________________________________________________________________________ Material submitted for another award I declare that no material contained in the thesis has been used in any other submission for an academic award and is solely my own work ____________________________________________________________________________ Signature of Candidate Type of Award Doctor of Philosophy (PhD) School Pharmacy and Biomedical Science, University of Central Lancashire 2 Abstract This study aims to design and develop effervescent proliposomes that could disintegrate in water and liberate liposomes, and to investigate the potential suitability of liposomes generated for aerosolization to target paranasal sinuses. Novel effervescent proliposomes prepared with Soya phosphatidylcholine (SPC) and Dipalmitoylphosphatidylcholine (DPPC) successfully generated stable liposomes with an improved disintegration time of less than 5 min. Differences in lipid composition were found to influence liposome size and drug entrapment of the hydrophobic drug Beclometasone dipropionate (BDP). Mannitol-based formulations developed with DPPC:Chol (1:1) produced liposomes of 7.54±0.15 µm with a drug entrapment efficiency of 82.15±8.29%. Addition of the mucoadhesives alginic acid or chitosan to effervescent proliposomes made with SPC was found to hamper BDP entrapment in liposomes. Effervescent proliposomes produced SPC:Chol liposomes that also proved beneficial for entrapment of the hydrophilic drug Xylometazoline hydrochloride (XH). The Pari Sinus (pulsating aerosol technology) and Pari Sprint (non-pulsating technology) nebulizers were used for liposome delivery to a nasal cast. Choice of carrier did not affect the liposome’s ability to withstand shearing. A novel system of a Sar-Gel® (water indicating paste) coated clear nasal cast fixed to a two-stage impinger system was set up to analyze drug deposition within the nasal cast cavity. Sinus drug deposition with effervescent mannitol, DPPC:Chol formulation was observed to be highest at 48.45±2.75 cm2 with pulsation compared to deposition of 35.52±11.11 cm2 without pulsation. Drug distribution studies indicated that the Pari Sinus deposited 10.47±2.9% drug, while the Pari Sprint deposited only 4.6±1.4%. The degree of drug loss was higher with conventional liposomes in the Pari Sinus nebulizer, indicating that the degree of bilayers disruption depended on formulation. 3 Table of Contents DEDICATION ............................................................................................................. 1 ABSTRACT ............................................................................................................. 3 TABLE OF CONTENTS ................................................................................................ 4 LIST OF TABLES ........................................................................................................... 8 LIST OF FIGURES ........................................................................................................ 9 ACKNOWLEDGEMENTS ......................................................................................... 13 LIST OF ABBREVIATIONS ....................................................................................... 15 CHAPTER 1 INTRODUCTION ........................................................................... 16 1.1. Nasal Drug Delivery .................................................................................................... 17 1.2. Anatomy of Nasal Cavity............................................................................................. 18 1.3. Para Nasal Sinuses (Sinuses) ....................................................................................... 21 Sinuses and sinusitis .............................................................................................21 Prevalence .............................................................................................................22 Anatomy of sinus ..................................................................................................22 Pathophysiology ...................................................................................................24 Complications of sinusitis.....................................................................................25 Treatment for sinusitis ..........................................................................................27 1.4. Nasal Drug Delivery Devices ...................................................................................... 31 Aerosol drug delivery to sinus ..............................................................................32 SinuNeb™ device .................................................................................................33 RinoFlow™ nasal aerosol delivery device .............................................................34 PARI Vibrent™ device .........................................................................................34 Pari Sinustar ™ device .........................................................................................35 Pari Sinus ™ device pulsating aerosol system .....................................................36 1.5. Liposomes .................................................................................................................... 40 An overview of liposomes and proliposomes .......................................................40 Historical background of liposomes .....................................................................42 Characterisation of liposomes ...............................................................................42 1.6. Advantages and Disadvantages of Liposomes ............................................................. 44 General advantages of liposomes .........................................................................44 Liposomes as drug carriers ...................................................................................44 Drawbacks of liposomes .......................................................................................46 Proliposomes ........................................................................................................46 Nasal delivery of liposomes ..................................................................................48 4 Nasal delivery of drugs through aerosolised proliposomes ..................................49 1.7. Effervescent Formulations for Nasal Drug Delivery ................................................... 50 Introduction to effervescent formulations .............................................................50 Effervescent liposomes .........................................................................................52 1.8. Model Drugs for the Study Couples with Effervescent Liposomes ............................. 54 Beclometasone dipropionate (hydrophobic) .........................................................54 Xylometazoline Hydrochloride (XH) ...................................................................55 1.9. Aim of this thesis ......................................................................................................... 57 1.10. Element of Originality: ............................................................................................ 59 CHAPTER 2 METHOD AND MATERIALS ....................................................... 60 2.1. Materials ...................................................................................................................... 61 2.2. Preparation of proliposomes ........................................................................................ 62 Preparation of particulate-based proliposomes .....................................................62 Sucrose based conventional liposomes (SPC based) with hydrophobic drug ......62 Mannitol based conventional liposome (SPC based) with hydrophobic drug ......63 Mannitol based conventional liposomes (DPPC based) with hydrophobic drug..64 Mannitol based conventional liposomes (SPC based) with hydrophilic drug ......64 2.3. Characterisation of liposomes ...................................................................................... 65 Particle size analysis: ............................................................................................65 Zeta potential (surface charge) analysis: ..............................................................65 Surface morphology using scanning electron microscopy (SEM) .......................65 2.4. Entrapment studies ....................................................................................................... 67 High –performance liquid chromatography (HPLC studies) for drug beclometasone dipropionate (BDP) ......................................................................67