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AIM To explore the potential of paclitaxel (PTX)-loaded anacardic acid conjugated hydrophobized gelatin nanoparticles. MATERIALS & METHODS Nanoparticles prepared by nanoprecipitation technique were evaluated for various quality attributes (particle size, % entrapment efficiency) in vitro drug release, MCF-7 cell uptake, cell cytotoxicity, in vivo pharmacokinetics, antitumor efficacy and toxicity. RESULTS The nanoparticles (250-300 nm, 74% entrapment efficiency) showed approximately 2.26-fold higher apoptosis index and approximately 5.86-fold reduction in IC50 value compared with PTX in MCF-7 cells. Also, approximately 3.51- and 1.36-fold increase in area under the curve compared with Intaxel® and Nanoxel™ (both from Fresenius Kabi, Gurugram, India) was achieved. Significant tumor burden reduction (∼60%) and reduced toxicity was observed compared with marketed formulations. CONCLUSION The hydrophobized gelatin nanoparticles displayed promising therapeutic potential, paving a new path for efficient PTX delivery.