AIM: To investigate F96, a N-acylethanolamine acid amidase (NAAA) inhibitor, as a novel drug for treating dry eye disease (DED) and to enhance its corneal retention time by utilizing nanometer micelles to improve therapeutic efficacy. METHODS: The study compared nanomicelles encapsulating doxorubicin with an aqueous solution of doxorubicin to assess the ability of the nanomicelles to prolong drug retention on the ocular surface. Dry eye was induced in mice through subcutaneous injections of scopolamine hydrobromide. The efficacy of F96 was evaluated using various clinical assessments, including the phenol red cotton test, Oregon green dextran staining, periodic acid-Schiff (PAS) staining, and Terminal dUTP Nick-End Labeling (TUNEL) assay. RESULTS: Doxorubicin micelles exhibited significantly prolonged retention compared to the aqueous solution. By 15min, the corneal fluorescence intensity of the micelle group was markedly higher than that of theaqueous solution group (P<0.05), and this enhanced effect persisted for at least 4h. Furthermore, mice treated with F96 demonstrated superior outcomes in tear production, corneal staining, and goblet cell density compared to the control groups. Specifically, F96-mPPP significantly increased tear secretion (3.35±0.45 vs 1.85±0.51 mm in the vehicle group, P<0.001), restored conjunctival goblet cell density (54.5±4.5 vs 31.3±3.0, P<0.01), and reduced corneal fluorescein staining scores (3.4±0.32 vs 6.5±0.72, P<0.001). Additionally, F96-mPPP treatment markedly decreased TUNEL-positive cells in the corneal epithelium, indicating suppression of apoptosis. CONCLUSION: F96 nanometer micelles have the potential to serve as a promising novel approach for effectively alleviating ocular surface damage in the treatment of dry eye disease.