Efficient logistics and transport infrastructure are critical in contemporary urban and interurban scenarios due to their impact on economic development, environmental sustainability, and quality of life. This study explores the use of the Ant Colony Optimization (ACO) metaheuristic applied to the Vehicle Routing Problem (VRP) and the strategic positioning of service stations along major highways. Through a systematic mapping of the literature and practical application to a real-world scenario—specifically, a case study on the Bandeirantes Highway (SP348), connecting Limeira to São Paulo, Brazil—the effectiveness of ACO is demonstrated in addressing complex logistical challenges, including capacity constraints, route optimization, and resource allocation. The proposed method integrates graph theory principles, entropy concepts from information theory, and economic analyses into a unified computational model implemented using Python (version 3.12), showcasing its accessibility for educational and practical business contexts. The results highlight significant improvements in operational efficiency, cost reductions, and optimized service station placement, emphasizing the algorithm’s robustness and versatility. Ultimately, this research provides valuable insights for policymakers, engineers, and logistics managers seeking sustainable and cost-effective solutions in transport infrastructure planning and management.