“PREPARATION AND EVALUATION OF

MICROSPHERES CONTAINING

AN ANTI-VIRAL DRUG”

SYNOPSIS FOR

M.PHARM DISSERTATION

SUBMITTED TO

RAJIV GANDHIUNIVERSITY OF HEALTH SCIENCES

KARNATAKA

BY

RUCHI KUMARI

I M.PHARM

DEPARTMENT OF PHARMACEUTICS

PESCOLLEGE OF PHARMACY

BANGALORE-560 050

(2012-2014)

1

RAJIVGANDHIUNIVERSITY OF HEALTH SCIENCES

KARNATAKA, BENGALURU

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. / Name of the candidate and address / RUCHI KUMARI
1st M. PHARM (PHARMACEUTICS)
PESCOLLEGE OF PHARMACY
HANUMANTHANAGAR
BENGALURU-560 050

PERMANENT ADDRESS:
C/O G.N.MAHARAJ
AT AMDANGA,GOURANDI ROAD
PO-ACHRA DISTRICT BURDWAN
WESTBENGAL-713335
2. / Name of the institution / PESCOLLEGE OF PHARMACY
HANUMANTHANAGAR
B.S.K.1st STAGE
BENGALURU:-560 050
3. / Course of the study / MASTER OF PHARMACY
(PHARMACEUTICS)
4. / Date of Admission / 11th July 2012
5. / Title of the topic:
“PREPARATION AND EVALUATION OF MICROSPHERES CONTAINING AN ANTI-VIRAL DRUG”
6. / Brief resume of the intended work:
6.1 Need for the study:
Viruses have the property to replicate very fast in host cell. It can attack any part of the host cell. Therefore, the clinical efficacy of antiviral drugs and its bioavailability is more important concern taken into account to treat viral infections. The oral and parenteral routes of drug administration have several shortcomings, which leads to formulating better delivery system.
Now, a day’s novel drug delivery systems (NDDS) proved to be a better approach to enhance the effectiveness of the antivirals and improve the patient compliance and decrease the adverse effect. The NDDS have reduced the dosing frequency and shorten the duration of treatment, thus, results in the treatment which is more cost-effective.
The development of NDDS for antiviral and antiretroviral therapy aims to deliver the drug devoid of toxicity, with high compatibility and biodegradability, targeting the drug to specific sites for viral infection and in some instances it also avoid the first pass metabolism effect. The usefulness of novel delivery approaches of antiviral agents such as niosomes, microspheres, micro emulsions, nanoparticles that are used in the treatment of various Herpes viruses and in human immunodeficiency virus (HIV) infections is very wide.
There arevarious 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. It is the reliable means to deliver the drug to the target site with specificity, if modified, and to maintain the desired concentration at the site of interest without untoward effects.
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. 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.
The present work therefore is planned to develop a controlled drug delivery system for an antiviral drug with microspheres as the carrier system.
6.2 Review of the literature
  • Hui Zhangsolidified IFN_-2b as microparticles firstly by co-lyophilizing it with gelatin and ZnSO4. Microspheres were then prepared.The preparing procedure and formulation were optimized with encapsulation efficiency and in vitro release as main parameters. Finally, the microspheres were prepared by S/O/W method with microparticle size about 5_mand PEGT/PBT-PLGA (9:1, w/w) as matrix material.In conclusion,the procedure and formulation used in this study were supposed to be successful to keep IFN_-2b active and released constantly and completely.1
  • S. Santoyo prepared poly(lactide-co-glycolide) (PLGA) microparticles bysolvent evaporation and spray-drying methods. Microparticles prepared by spray-drying showed a encapsulationefficiency of 80%. Conversely, for all the microspheres prepared by the W/O/W solvent evaporation method the
encapsulation efficiency was low. The results obtained showed that the amount ofcidofovir penetrated, over a 24 h time period, was higher with the drug solution than with microparticles.The profiles obtained for the two formulations showed that the quantity of cidofovirretained in the skin decreased with the depth. Besides the amount of cidofovir found in the basal epidermis (120–150m) was much higher with microparticles than with the control solution.2
  • Sandra Kockisch develop mucoadhesive microspheres that can be utilised for the controlled release of triclosan in oral-care formulations, specifically dental pastes. Triclosan was rapidly released into a sodium lauryl sulphate containingbuffer from all but the chitosan microspheres. The release of triclosan from microspheres suspended in a non-aqueous paste, was found to be sustained over considerable time-periods, which were influenced strongly by the nature of the polymeric carrier.3
  • S.Govender experimental design was employed to statistically optimise the formulation parameters of a tetracycline microsphere preparation for maximum bioadhesivity and controlled drug release. Thermal analyses showed a possible interaction between the drug and polymer. Scanning electron microscopy confirmed the integrity of the microspheres and identified the morphological changes following drug release. Surface pH of the microspheres was similar to salivary pH and did not show extremes in changes over the test period.4
  • Elisabetta Esposito prepared and characterized Ca-alginate microspheres obtained by a new procedure from alginic acid,treated with an excess of NaOH and subsequently with a very concentrated solution of CaCl2. The produced particles have a spherical shape with an average diameter of 350 mm. The particles show a relative dense and homogeneous internal structure in comparison with previously reported alginate microcapsules (the bulk densitybeing 0.47 g/cm3), consequently the particle matrix is characterized by a high density of charged carboxylic groups.Particles produced with a contact time of 1 h showed a degree of crosslinking of 54%.5
  • M. Elorza developed micro-particles (MPs) and nano-particles (NPs) containing ACV have been developed using cross-linked chitosan with tripolyphosphate (TPP) due to the biocompatibility, bio-adhesion ability and the potential power as penetration enhancer of this polymer. From the diffusion profiles, it was found that the amounts of ACV effectively diffused in 24 h were 30, 430 and 80 g for the ACV solution,MP and NP respectively.6
  • M.D. Abd El-Hameed prepared microspheres of hydrophilic polymers were by the w/o emulsification solvent evaporation technique with a potential application as drug carriers for nasal administration by insufflation..The FITC-dextran was released from the microspheres initially at a constant rate. However the release rate subsequently decreased over the 24 h test period. No differences were observed for release from Carbopol 934P, PVA and HPMC: all exhibited faster release than that achieved from the Chitosan microspheres which exhibited a size-dependent release effect.7
  • Tze-Wen Chung prepared with O:W emulsion–solvent evaporation process. Prepared in the fast rate of solvent evaporation (FRSE) process by reducing ambient pressure, smoothly morphological surface of drug loaded PLLA and PDLLA microspheres was observed. While in the normal rate of solvent evaporation (NRSE) process, roughness or pinhole surface was only found at drug loaded PLLA microsphere.8
  • V. Carelli prepared NAM-medicated PCP in the 4:1 PCP-NAM wt ratio and dispersed, at the 20% or 40% concentration, in silicone in the form of osmotically active particles of around 15 mm mean volume diameter, and encapsulated in microspheres in the 105–710 mm size range by a modified emulsion vulcanization technique, with 100% entrapment efficiency. The external and internal morphology of microspheres, and the size distribution of PCP-NAM particles dispersed therein were evaluated by scanning electron micrography.9
  • WeienYuan developed anovel protein drug sustained-release system, which
is based on the combination of protein-loaded dextran and PLGA
microspheres.The dextran glassy particles can stabilize proteins in the
PLGA matrix, which is the major advantage of this novel protein sustained
release system over preparation for the PLGA microspheres using
W/O/W double-emulsion method.10
  • Xinjian Cheng produced a core–shell-structured laminar liquid jet, which breaks to form monodisperse compound liquid droplets. Stabilized in a dilute aqueous polyvinyl alcohol (PVA) solution, the droplets are converted into solid protein-loaded polymer microspheres through evaporation of theorganic solvent. Results showed that preparation parameters like polymer concentration,total flow rate,flow rate ratio of the aqueous to organic have significant effects on the mean particle size, particle morphology and protein encapsulation efficiency (EE).11
  • Xinming Liu prepared a novel microsphere drug delivery system of ivermectin using hydrophobic protein zein by the phase separation method and characterized by a scanning electron microscope and laser light scattering particle size analyzer. Release of model drug IVM from zein microspheres, tabletted microspheres and pepsin degradation of tabletted microspheres were also performed in vitro to investigate the mechanism of model drug release. The results showed that the zein microspheres and tabletted microspheres are suitable for use as a sustained-release form of IVM.12
  • Anil K.anal produced pentasodium tripolyphosphate cross-linked chitosan smicrospheres with higher acid resistance for controlled release of ampicillin. The microspheres were prepared by two different microencapsulation procedures (by emulsification and by spray-drying) and characterized by their particle size, surface morphology, stability, drug entrapment efficiency and drug release The cross-linked microspheres were more stable in simulated gastric fluid and showed slower but sustained release of ampicillin. The antimicrobial activity of the released ampicillin was confirmed by Staphylococcus aureus bioassay.13
  • Edith J.A.M. Schlicher developed a controlled release system for the water- soluble drug DFO based on poly (D,L-lactic-co-glycolic acid) (PLGA). PLGA microspheres containing DFO were prepared using the water-in-oil-in-water (w:o:w) solvent evaporation technique .DFO was released from the microspheres in an initial burst (controlled by formulation parameters) followed by marginalrelease and a pulse release at the time the microspheres start to dissolve (20–30 days). The release of DFO was incomplete after complete dissolution of the microspheres. This is probably caused by the instability of DFO at lowpH, generated upon degradation of the PLGA matrix.14
  • E.Gde Jalo developed acyclovir loaded microparticles, prepared by an O/W solvent evaporation method. Their antiviral activity against herpes simplex virus type 1 (HSV-1) and toxicity were evaluated on Vero cells and then compared with those presented by a drug solutionacyclovir-loaded microparticles have shown to be promising carriers for the effectivedelivery of acyclovir in the treatment of HSV-1 infections in cells so they can have a potential use in vivo.15
  • Ganza-Gonza prepared microspheres were by spray drying of aqueous polymer dispersions containing the drug and different amounts of formaldehyde as cross-linker. Drug release kinetics were investigated in vitro in media of different pH. Chondroitin sulphate microspheres scarcely retarded drug release, regardless of cross-linker concentration and medium pH, and were thus not further characterized Electron microscopy showed the chitosan microspheres to be almost-spherical, though with shallow invaginations.16

6.3 Main objectives of the study:
The objectives of the present study is as follows:
a. To carry out the preformulation studies for the drug and the excipients.
b. To formulate microspheres consisting of an antiviral drug using the selected polymers.
c.To evaluate the microspheres for entrapment efficacy and by dissolution studies.
d. To optimize the prepared microspheres based on the in-vitro release profiles.
e.To carry out the stability studies of the selected formulation.
7 / 7.1Source of data
1.The data will be obtained from literature survey and internet source.
2.The data will be obtained on experiment work
a)Formulation of the microspheres
b)In vitro evaluation of the formulations
c)Stability studies of microspheres.
7.2 Method of collection of data (including sampling procedures if any)
The data will be collected from the formulation, in-vitro evaluation, and stability studies.
7.3Does the study require any investigation or interventions to be
Conducted on patients or other humans or animals?
NO
7.4 Has ethical clearance been obtained from your institution in case of
7.3?
NOT APPLICABLE
8 /

List of References:

1.Zhiping L, Yan L, Zhanga H, Xueru L, Luoa F, X M. Development of interferon alpha-2b microspheres with constant release. Int J Pharm. 2011;410:48-53.
2.Santoyo S, Blanco MJ, Ygartua P, Renedo MJ. Optimization of topical cidofovir penetration using microparticles. Int J Pharm. 2002;242:107-13.
3.Kockischa S, Gareth D, Tsibouklisc J, D. J. Mucoadhesive, triclosan-loaded polymer microspheres for application to the oral cavity: preparation and controlled release characteristics Euro J Pharm and Biopharm. 2005;59:207-16.
4.Govender S, Pillay V, Chetty DJ, Essack SY, Dangora GM, T G. Optimisation and characterisation of bioadhesive controlled release tetracycline microspheres Int J Pharm. 2005;306:24-40.
5.Fundueanu G, Esposito E, Mihai D, Carpov A, M R, NC. Preparation and characterization of Ca-alginate microspheres by a new emulsification method. Int J Pharm. 1998;170:11-21.
6.Calderon L, Harris R, Cordoba M, Elorza M, Elorza B, D C. Nano and msicroparticulate chitosan-based systems for antiviral topical delivery. Euro J PharmaSci. 2013;48:216-22.
7.Abd El-Hameed MD, IW K. Preparation and in vitro characterisation of mucoadhesive polymeric microspheres as intra-nasal delivery systems
Euro J PharmaSci
1997;44:53-60.
8.Wen Chung T, YouHuang Y, Y L. Effects of the rate of solvent evaporation on the characteristics of drug loaded PLLA and PDLLA
Microspheres Int J Pharm. 2001;212:161-69.
9.Carelli V, DiColo G, Gesi M, Martini F, E N. Mechanism of drug release from silicone microspheres containing Polycarbophil Int J Pharm. 1997;153:105-1.
10.Weien Yuan, Wu F, Guo M, T J. Development of protein delivery microsphere system by a novel S/O/O/W multi-emulsion. Euro J Pharm sci. 2009;36:212-18.
11.Cheng X, Liu R, H Y. A simple method for the preparation of monodisperse protein-loaded microspheres with high encapsulation efficiencies
Euro J Pharm and Biopharm. 2010;76:336-41.
12.Liua X, Sunb Q, Wangb H, J Y. Microspheres of corn protein, zein, for an ivermectin drug delivery system. Journal of Biomaterials. 2005;26:109-15.
13.Anil K, Willem F, C R. Ionotropic cross-linked chitosan microspheres for controlled release of ampicillin. Int J Pharm. 2006;312:166–73.
14.Edith JS, Nancy S, Zuidema J, Talsma H, E H. Preparation and characterisation of Poly (D,L-lactic-co-glycolic acid) microspheres containing desferrioxamine. Int J Pharm. 1997;153:235-45.
15.Gade E, Blanco MJ, Ygartuaa P. Increased efficacy of acyclovir-loaded microparticles against herpes simplex virus type 1 in cell culture. Euro J of Pharm and Biopharm. 2003;56:183-87.
16.Ganza-Gonza.A, Anguiano-Igea.S, Otero-Espinar.FJ, MeÂndez.J B. Chitosan.1999;48:149-55.
9.

10.

11.

12. /
Signature of the candidate:
(Ruchi Kumari)
Remarks of the guide: RECOMMENDED
11.1Name And Designation of Guide: Dr.S.VASANTI
AssociateProfessor,
Department of Pharmaceutics,
P.E.SCollege of Pharmacy,
Bengaluru-50.
11.2 Signature
11.3 Co-Guide NOT APPLICABLE
11.4 Signature
11.5 Head of the department Recommended and Forwarded by
C.S.Satish
Professor Head,
Department of Pharmaceutics,
P.E.SCollege of Pharmacy,
Bengaluru-50.
11.6 Signature
12.1 Remarks of the Chairman and Principal:
Recommended and Forwarded by
Prof. Dr. S. Mohan
Principal and Director,
P.E.S College of Pharmacy,
Bangalore-50.
12.2 Signature.

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