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INTRODUCTIONOndansetron HCl (OND) is an antiemetic drug used for preventing and treatment of nausea and vomiting induced by chemotherapy, radiotherapy and anesthesia. It has and intravenous dosage forms are commercially available. However following the oral administration oral bioavailability is nearly 60% due to first pass metabolism. Also it has relatively short half life therefore drug needs to administrate several times during treatment[1]. The aim of this study was to prepare OND loaded polymeric nanoparticles(NPs) to extend the residence time of the drug in circulation to reduce dosing frequency and to increase patient compliance.MATERIALS AND METHODSMaterialsPLGA (50:50) (Resomer®RG 502H), PVA (MW 31–50 kDa, 87–89% hydrolyzed), dialysis membrane (MW: 14kDa), the organic solvents, dichloromethane (DCM) and acetonitrile (ACN) were purchased from Sigma-Aldrich (St. Louis, MO). OND was a gift from Adeka Pharmaceutical (Istanbul, Turkey).MethodPreparation and characterization of NPsOND loaded polymeric NPs were prepared by double emulsion solvent evaporation method[2]. The optimum formulation was selected among the formulations in our previous study and pegylated[3]. Formulation codes and composition are given Table1.The formulations were characterized in terms of PS, PDI, ZP, EE% and in vitro drug release.The lyophilized nanoparticles were dissolved in acetonitrile to determine of encapsulation efficiency (EE%). The encapsulated drug after dilution with the mobile phase was determined using the HPLC method.Compatibility studies of drug and excipientsThe possible interactions between drug and the components of NPs by Differential scanning calorimetry (DSC) and Fourier-Transform Infrared Spectroscopy (FTIR).Surface morphologiesThe surface characteristics of the OND loaded nanoparticles were investigated by Transmission electron microscopy (TEM). Cytotoxicity studies (MTT assay)The cytotoxicity effect of the nanoparticles was tested by MTT viability assay using the L929 cell line.RESULTS AND DISCUSSIONParticle size, particle size distribution zeta potential and encapsulation efficiencyThe average particle size of NP4 and PEG NP4were found 342 ± 6,66 nm and 341 ± 7,25nm, respectively. The EE% values of NP4 and PEGNP4 were obtained 46,8 ± 2,78 % and 47,8 ± 1,69 %, respectively.It was observed that there was no significant difference in particle size, PDI and EE% values between NP4 and PEG NP4 formulations (p>0.05). It has been observed that the PDI of nanoparticles is less than 0.3, which indicated that homogenous populations of the nanoparticles. The zeta potential value of PEG NP4 was slightly more negative compared to the uncoated nanoparticles (NP4). The overall negative value of zeta could attributed to negative charge of PLGA.Compatibility studies of drug and excipientsAs a result of DSC and FTIR analysis, no interaction was observed between the drug and excipients. In the nanoparticle formulation, the drug peak disappeared, which was confirmed that the drug was molecularly dispersed inside of the NPs.Transmission electron microscopy (TEM)The TEM image of NP4 and PEGNP4 revealed that the NPs were in the nano-range. They were fairly spherical in shape. The PEG layer was clearly sighted in the TEM images of the PEGNP4 formulation.In vitro drug release studyThe percent cumulative release of OND loaded nanoparticle formulations in phosphate buffer solution pH 7.4 is shown in Figure 4. At the end of the 72 h, 91,6 and 92,7% OND was released from NP4 and PEGNP4 respectively.Cytotoxicity studies (MTT assay)The biocompatibility and viability were assessed using the L929 cell line. These cells were incubated with free OND, Zofer, NPs without drug, OND loaded NPs and analyzed for their viability at 24 and 48h. PEGylated NPs has been found within acceptable range and were seen to be nontoxic in the L929 cell. These results revealed that NPs should be considered as a safe drug delivery system.CONCLUSIONOND loaded polymeric NPs were developed successfully. The nanoparticles had a good sphericity, a uniform distribution of particle size and non-cytotoxic nature. NP4 andPEGNP4 formulations exhibited a sustained-release characteristic in vitro. Also it wasachieved high encapsulation efficiency value although OND is hydrophilic nature. These findings are quite encauraging for future in vivo studies.ACKNOWLEDGEMENTThis work has been supported by Gazi University Scientific Research Projects Coordination Unit under grant number TCD2021-6957 |
