Glaucoma is a chronic, gradual and very serious neurodegenerative disease, and its natural course is blindness.In fact, it is the leading cause of irreversible blindness in the world. Current treatments have several limitations,such as limited bioavailability and low protection of the optic nerve. Therefore, in this work the development of a formulation containing second-generation lipid nanoparticles, known as nanostructured lipid carriers (NLCs),loaded with a natural flavonoid poorly soluble in water has been undertaken. For the synthesis of these NLCs, the hot high-pressure homogenization method has been used, where a high amount of energy is applied. Using the optimized formulation, the HET-CAM test was performed, an in vitro ocular tolerance test on embryonic eggs,classifying the formulation as practically non-irritating.