Microemulsions (ME) have gained attention as an alternative pharmaceutical formulation for transdermal delivery systems. However, the complicated relationships between various ME compositions (causal factors) and their characteristics (response variable) have not been fully comprehended. To overcome this problem, the design and development of ME for transdermal delivery was performed in our study using Design Expert® Software. The model formulations of ME were prepared according to the ME region obtained from pseudo-ternary phase diagrams using the simplex lattice design as an optimization technique. In this study, ketoprofen-loaded ME composed of oleic acid, Cremophor® RH40, ethanol and water were prepared, and their characteristics ( e.g. , size, charge, conductivity, pH, viscosity, drug content, loading capacity and skin permeation flux) were evaluated. The ME having an appropriate skin permeation flux was used as the basis for optimization. The skin permeation flux of the experimental ME was very close to the flux predicted by Design Expert® Software and was significantly greater than that for the commercial product. Possible mechanisms for the enhancement of the skin permeation of the ME were also investigated using Fourier transform infrared (FT-IR) spectroscopy and X-ray diffraction (XRD). This finding provided an understanding of the relationship between the causal factors and response variables, as shown in the response surfaces. Moreover, these results indicated that the simple lattice design was beneficial for the pharmaceutical development of ME for transdermal delivery.