The objective of this study was the development and validation of an integrated measure-assessment approach (driving performance and psychophysiological indicators) for the assessment of driving demands of rural road segments as a starting point for design. With 31 student test drivers and a selection of six study roads, the reactivity of psychophysiological parameters was evaluated based on a general model of cognitive-energetic effort regulation according to Hockey (1993). The road curvature change rate was introduced as the independent variable, which served as a criterion of objective road difficulty. Based on a reliability analysis, the longitudinal and cross-sectional stability, especially for heart rate (HR) and blink rate (BR) measures, is sufficiently high. Both these psychophysiological variables and speed vary as a function of the curvature change rate of the rural road segments. The speed parameter differentiates very strongly the different curvature change rates. Among the psychophysiological indicators, changes in the blink rate almost exactly mirror the level of the curvature change rate owing to the fact that the BR decreases almost steadily with increasing curvature change rate. Cardiovascular parameters, such as heart rate (HR) and heart rate variability (HRV), and skin conductance response (SCR) also vary in relation to driving difficulty of road segments, but at levels of intermediate curvature other additional road characteristics may also influence these measures.