Background and Aims: Invasive pulmonary aspergillosis (IPA) is an imperative concern in the present era due to its high occurrence and mortality rate in severely immunocompromised patients. The present study was designed to develop, optimize and characterize encapsulated nystatin (NYS) cubosome nanoparticles as an inhalable system, a viable alternative for effective management of IPA. Methods: A dry lipidic film comprising glycerol monooleate (GMO), Span 83, Poloxamer (P-407) and dispersed NYS was subjected to ultrasound sonication to produce colloidal dispersion of cubosomes. The process and formulation variables were screened using Plackett Burman design and further optimized by Box Behnken design by evaluating its effect on particle size, polydispersity index (PDI), zeta potential and entrapment efficiency. Results: The optimized NYS cubosomes were nearly spherical with some irregular polyangular symmetry as visualized by transmission electron microscopy (TEM). Further, small angle X-ray scattering (SAXS) affirmed Pn3m cubic mesophasic structure. The optimized nanoparticles had particle size 263.5 nm, zeta potential -14.4 mV, PDI 0.283 and entrapment efficiency 82%. The in-vitro cytotoxicity assay indicated that NYS cubosomes reduced cell cytotoxicity in contrast to pure drug post 48h. In-vitro haemolytic assay denoted lower toxicity of formulation as compared to free drug. In-vitro drug release studies highlighted, slow but continuous release from NYS cubosomes until 48h and showcased Higuchi release kinetics. Likewise, NYS cubosome demonstrated higher antifungal activity compared to drug suspended in phosphate buffer. Conclusion: Thus, non-invasive feature and contemplated target specificity of nystatin loaded cubosome nanoparticles pave a mode for its prospect as pulmonary delivery to combat IPA.
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