Development and Characterization of Polymeric Nanoparticulate Delivery System for Hydrophillic Drug: Gemcitabine.

dc.contributor.advisorDash, Alekha K.en_US
dc.contributor.authorKhurana, Jatinen_US
dc.contributor.cuauthorKhurana, Jatinen_US
dc.date.accessioned2010-03-09T23:28:08Z
dc.date.available2010-03-09T23:28:08Z
dc.date.issued2009-12en_US
dc.degree.committeeShara, Michaelen_US
dc.degree.committeeSingh, Somnathen_US
dc.degree.disciplinePharmaceutical Science (graduate program)en_US
dc.degree.grantorGraduate Schoolen_US
dc.degree.levelMS (Master of Science)en_US
dc.degree.nameM.S. in Pharmaceutical Sciencesen_US
dc.description.abstractGemcitabine is a nucleoside analogue, used in various carcinomas such as non small cell lung cancer, pancreatic cancer, ovarian cancer and breast cancer. The major setbacks to the conventional therapy with gemcitabine include its short half-life and highly hydrophilic nature. The objectives of this investigation were to develop and evaluate the physiochemical properties, drug loading and entrapment efficiency, in vitro release, cytotoxicity, and cellular uptake of polymeric nano-particulate formulations containing gemcitabine hydrochloride. The study also entailed development and validation of a high performance liquid chromatography (HPLC) method for the analysis of gemcitabine hydrochloride.|A reverse phase HPLC method using a C18 Luna column was developed and validated. Alginate and Poly lactide co glycolide/Poly-ɛ-caprolactone (PLGA:PCL 80:20) nanoparticles were prepared by multiple emulsion-solvent evaporation methodology. An aqueous solution of low viscosity alginate containing gemcitabine was emulsified into 10% solution of dioctyl-sulfosuccinate in dichloro methane (DCM) by sonication. The primary emulsion was then emulsified in 0.5% (w/v) aqueous solution of polyvinyl alcohol (PVA). Calcium chloride solution (60% w/v) was used to cause cross linking of the polymer. For PLGA:PCL system, the polymer mix was dissolved in dichloromethane (DCM) and an aqueous gemcitabine (with and without sodium chloride) was emulsified under ultrasonic conditions (12-watts; 1-min). This primary emulsion was further emulsified in 2% (w/v) PVA under ultrasonic conditions (24-watts; 3-min) to prepare a multiple-emulsion (w/o/w). In both cases DCM, the organic solvent was evaporated (20- hours, magnetic-stirrer) prior to ultracentrifugation (10000-rpm for PLGA:PCL; 25000-rpm for alginate). The pellet obtained was washed thrice with de-ionized water to remove PVA and any free drug and re-centrifuged. The particles were re-suspended in de-ionized water and then lyophilized to obtain the dried powdered delivery formulation. Particle size and surface charge of the nano-particles were measured using zeta-sizer. The surface morphology and microstructure were evaluated by scanning electron microscopy The drug loading and entrapment efficiencies were evaluated by a HPLC method (Luna C18 column (4.6 X 250 mm), 95/5 (v/v) 0.04M ammonium acetate/acetonitrile mobile phase (pH 5.5), 1.0 ml/min flow rate and 268 nm UV detection). Differential scanning calorimetry (DSC) was used to determine the physical state of gemcitabine in the nanoparticles. The cytotoxicity in pancreatic cancer cells (BxPC-3) was evaluated by MTT assay. The cellular uptake of gemcitabine solution and gemcitabine loaded alginate nano-particle suspension in BxPC-3 cells was determined for 15, 30 and 60 minutes.|The particle-size and surface-charge was 564.7±56.5nm and -25.65±1.94mV for PLGA:PCL and 210.6±6.90nm and -33.21±1.63mV for alginate. Both the nano-particles were distinctly spherical and non-porous. The drug load was 5.14% for PLGA:PCL and 6.87% for alginate-particles, and the practical entrapment efficiency was found to be 54.1 % and 22.4% respectively. However, in case of PLGA:PCL particles, a two-fold increase in the entrapment efficiency was observed with the addition of sodium-chloride. The absence of endothermic melting peak of the drug in the DSC thermogram was an indication of the non-crystalline state of gemcitabine in the nanoparticles. In addition, there was no cytotoxicity associated with nanoparticle concentrations at-or-below 5mg/mL. The uptake of nano-particles was around 4 times higher than the solution with treatment for 15 minutes and increased to almost 7 times following treatment for 60 minutes.|Gemcitabine hydrochloride could be successfully formulated into a sustained release nano-particulate formulation using calcium cross-linked alginate and dioctyl sulfo succinate system. The nano-particulate delivery system exhibited better cytotoxic activity and also significantly enhanced the accumulation of the drug in BxPC-3 cell monolayers.en_US
dc.description.pagesxxi, 100 pagesen_US
dc.identifier.otherKhurana-thesis.pdf
dc.identifier.urihttp://hdl.handle.net/10504/5531
dc.language.isoen_USen_US
dc.publisherCreighton Universityen_US
dc.publisher.locationOmaha, Nebraskaen_US
dc.rightscopyright 2009, Jatin Khuranaen_US
dc.rights.holderJatin Khuranaen_US
dc.subject.meshDeoxycytidine--analogs & derivativesen_US
dc.subject.meshDrug Delivery Systemsen_US
dc.subject.meshChromatography, High Pressure Liquid--methodsen_US
dc.titleDevelopment and Characterization of Polymeric Nanoparticulate Delivery System for Hydrophillic Drug: Gemcitabine.en_US
dc.typeThesis
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