The invention discloses a multi-user real-time path planning method and system based on ant colony algorithm.The multi-user real-time path planning method comprises the steps that S1, parameters for performing path optimization are initialized; S2, the path optimization is carried out by the improved ant colony algorithm to obtain the optimal path; the improved ant colony algorithm comprises the steps that a flow factor is introduced for pheromone concentration update calculation when the path optimization is carried out. According to the multi-user real-time path planning method and system based on the ant colony algorithm, the path planning problem of vehicles under the complex road network of urban traffic can be better coped with by adding the flow factor based on the existing algorithm, so that each vehicle can arrive at the destination in shorter time without affecting the driving of other vehicles.