Abstract Due to the differences of physical and fluid properties in vertical oil layers, the interlayer contradictions are prominent and affect the reservoir utilization degree seriously. Therefore, the coordination between reasonable bottom-hole flow pressure and optimal production, as well as the reduction of interlayer interference are the primary concerns during the co-production of multi-layer oil reservoirs. Based on the reservoir engineering and fluid mechanics in porous media theory, this work adopt the comprehensive pressure system and consider the interlayer interference to establish a multi-branch horizontal well productivity model and interlayer interference mathematical model. By analyzing the main controlling factors and interference mechanisms, this article establishes a pattern of interlayer interference, and improves the quantitative characterization interlayer interference theory in multi-layer combined mining. The study has shown that: (1) The interlayer interference is beneficial for balancing the production of different layers and improving development efficiency, it is greatly affected by interlayer heterogeneity; (2) When the number of layers exceeds 2 layers, the interference coefficient increases; With the increase of the layer thickness, the thicker oil layers have higher productivity, and the thickness of layer has a significant effect on the production of low-pressure layer; When the production pressure difference increases, the interlayer interference can be reduced; (3) The interlayer interference mathematical model constructed in this paper has high prediction accuracy and strong practicality, which has theoretical guidance significance for the division of strata in comprehensive adjustment of reservoirs.