Low-temperature cracking is a prominent disease form of asphalt pavement, and the formation of early cracks mainly occurs at the interface of the mixture. In order to gain a deeper understanding of the microscopic interfacial behavior of asphalt mixtures cracking at low temperatures, an asphalt–aggregate interface model and an asphalt–asphalt interface model were developed through molecular dynamics simulations. Tensile simulations were carried out at six different temperatures of 0 °C, − 10 °C, − 20 °C, − 30 °C, −40 °C and − 50 °C. Using the interfacial stress as an evaluation index, the change in interfacial strength under different low-temperature conditions was investigated and compared with the macroscopic test results. The results show that the molecular dynamics (MD) simulations are in high agreement with the macroscopic tests. The interaction and damage behavior of the asphalt–asphalt interface are more complex than that of the asphalt–aggregate interface. The interfacial strength of the two models decreases with the decrease of temperature. The temperature sensitivity of the asphalt–asphalt model is higher. In the range of 0 °C to − 50 °C, adhesion failure between asphalt and aggregate predominates, while after − 50 °C, cohesive failure between asphalt is more likely to occur. The macroscopic test results show that the gradation, void ratio and asphalt content have significant effects on the low-temperature cracking resistance of the mixture.