Road alignment, traffic flow, and sign information interact and create complex traffic situations. To evaluate the effects of freeway horizontal radius, slope grade, traffic flow, and sign information on subjective driving workload and performance, a simulated driving experiment based on an orthogonal test design was conducted. The National Aeronautics and Space Administration Task Load Index was used to measure self-reported workload. Data regarding speed and lane deviation were collected through the driving simulator. A multivariate analysis of variance results indicated that the radius of the horizontal curve significantly influenced workload score, average speed, and lane deviation. It was observed that in a high workload environment, participants exerted more effort on the driving task; however, driving performance still decreased. Although there was no significant correlation between slope grade and subjective driving workload or performance, the primary effect of slope grade on workload and average speed was statistically significant. The amount of sign information significantly influenced the driver's perceived workload; however, it did not significantly impact driving performance. In addition, the low correlation coefficient between subjective workload and performance was obtained. These research findings can provide insights for the design of freeway alignments and traffic signs to maintain optimal workload and minimize safety risks.