This study aimed to develop gemcitabine-loaded nanocapsules for pancreatic cancer treatment, optimizing their formulation before evaluating them through in vivo studies. The researchers selected gemcitabine as the model drug because it has established clinical relevance and known pharmacokinetic shortfalls (short half-life and poor bioavailability) with toxic dose limits which suit the evaluation of nanoparticle drug delivery systems. The researchers conducted a 3 factorial design to optimize the formulation through adjustments of PLGA concentration and Tween 80 concentration. The optimal characteristics of F5 among nine formulations included particles of 160 ± 3 nm with 87 ± 2% encapsulation efficiency and - 27.8 ± 1.2 mV zeta potential. The in vitro drug release testing alongside pharmacokinetic results established that nanoparticle gemcitabine caused extended drug delivery and dramatically better bioavailability in addition to achieving longer systemic circulation than free gemcitabine alone. Evaluation of biodistribution revealed that the product displayed tumor-targeting property and had improved antitumor effect and less side effect compared with the other groups. The findings demonstrate that gemcitabine works as an ideal model chemotherapy drug to measure nanocarrier delivery platforms for treating pancreatic cancer solid tumors.