Dolutegravir sodium is an effective antiretroviral medication used for the treatment of HIV. The drug is classified as BCS class II, indicating its low solubility at the typical gastric pH. The purpose of this research was to identify and optimize the key operational and formulation factors affecting the drug’s properties, using a 32-factorial design. Nanosuspensions were prepared using PVP K30, polyvinyl alcohol, polxammer 407, and polxammer 188. The study focused on polymer concentration as the formulation variable and homogenization speed as the manufacturing variable. Particle size and saturation solubility were the main parameters evaluated. The analysis of variance showed that both the polymer concentration and homogenization speed significantly affected the nanosuspension’s particle size and saturation solubility. The results, including contour plots and polynomial equations, provide useful insights into the optimization of the independent variables for the formulation of nanosuspensions with the desired properties.