Efforts to improve the oral bioavailability of cyclosporine A (CyA) remains a challenge in the field of drug delivery. In this study, glyceryl monooleate (GMO)/poloxamer 407 cubic nanoparticles were evaluated as potential vehicles to improve the oral bioavailability of CyA. Cubic nanoparticles were prepared via the fragmentation of a bulk GMO/poloxamer 407 cubic phase gel by sonication and homogenization. The cubic inner structure formed was verified using Cryo-TEM. The mean diameters of the nanoparticles were about 180 nm, and the entrapment efficiency of these particles for CyA was over 85%. The in vitro release of CyA from these nanoparticles was less than 5% at 12 h. The results of a pharmacokinetic study in beagle dogs showed improved absorption of CyA from cubic nanoparticles as compared to microemulsion-based Neoral^®; higher C_max (1371.18 ± 37.34 vs 969.68 ± 176.3 ng mL⁻¹), higher AUC_0–t (7757.21 ± 1093.64 vs 4739.52 ± 806.30 ng h mL⁻¹) and AUC_0–∞ (9004.77 ± 1090.38 vs 5462.31 ± 930.76 ng h mL⁻¹). The relative oral bioavailability of CyA cubic nanoparticles calculated on the basis of AUC_0–∞ was about 178% as compared to Neoral^®. The enhanced bioavailability of CyA is likely due to facilitated absorption by cubic nanoparticles rather than improved release.