Synthesis, Characterization and binding studies of Polymeric Nanoparticles using Gemcitabine Hydrochloride

Abstract

Gemcitabine is a clinically valuable drug delivered intravenously. In order to explore other routes of administration for more efficacious drug delivery, its redevelopment for application through oral route with the help of nanotechnology is an ongoing thrust area. Nanotechnology helps the drug enter into tissues at the molecular level, with increased drug localisation and cellular uptake, larger surface area with modifiable biologic properties, mediate molecular interactions and identify molecular changes. The objective of the study was to use Eudragit RS100 to prepare polymeric nanoparticles of gemcitabine (GEM) in order to improve its half-life, reduce dosage and increase the stability of the drug. GEM polymeric nanoparticles were prepared by nanoprecipitation technique. They were characterized for particle size, zeta potential (ZP), drug content, entrapment efficiency (EE) and in-vitro drug release. Further, they were also evaluated using TEM, DSC and FTIR spectroscopy. Mechanistic insights of the synthesized nanoparticles were explored using a protein binding study, electrophoretic mobility shift assay (EMSA) and plasma protein binding study. Docking study was carried out to check the binding of the drug and polymer with DNA and protein. The synthesized GEM polymeric nanoparticles showed particle size in the range of 200-450 nm. Due to physical stability issues, optimized polymeric nanoparticles of GEM were lyophilized and exhibited a zeta potential of +11.9 mV, drug content 96.74% w/v and EE of 68-75% w/v. In-vitro drug release study demonstrated sustained release. Protein binding study with bovine serum albumin (BSA) revealed protein binding of GEM-loaded polymeric nanoparticles comparable with the marketed formulation (Oncogem 200, Cipla Ltd.). In addition to this, human plasma protein binding studies showed negligible interaction of GEM with plasma proteins with both formulations. EMSA displayed binding with CT-DNA. Lyophilized GEM nanoparticles were found to be stable and the mechanistic studies found them comparable to that of marketed formulation.