Naegleria fowleri is a free-living amoeba that causes primary amoebic meningoencephalitis. Despite combination drug therapies, N. fowleri is not sensitive to current drug therapies, contributing to the pathogen’s mortality rate of 98%. To enable rational drug designing, this study has proposed an integrated track of nanotechnology coupling with the enrichment concept. In the current study, zinc oxide nanoparticles (ZNP) were screened against ERK protein, which is responsible for the production of pro-inflammatory cytokines that cause brain disturbance in N. fowleri infection. Furthermore, an enrichment analysis has been executed to increase the efficiency of the ZNP through the addition of two amines and one chlorine group. The computational prediction of zeta potential, cytotoxicity, organ toxicity, calculations of binding free energy, and ADMET analysis shows that it is stable and possesses no toxic effect. Amine + chlorine enriched ZNP resulted in a binding energy of −7.8 kcal/mol, a zeta potential reliability of −40 mV, a cytotoxicity of −0.0002, inactive against all the targeted organ models, ADMET profiling shows a molecular weight of 320.54 g/mol, a lipophilicity of −0.99, high water solubility, and good gastrointestinal tract absorption. This proposed invention represents the future work for in vitro in combating this devastating disease toward a reliable therapeutic target with drugs that specifically aimed to inhibit the infection.