To meet the strict requirement on whole chain of vaccine production, storage, transportation, and distribution, most researches have been done on cascade refrigeration systems to achieve high operation performance, while a research gap on the performance exploration of eco‐friendly refrigeration systems still exists. In this paper, a comprehensive thermodynamic model was established to analyze the operation performance of a pre‐cooled cascade refrigeration system with eco‐friendly refrigerants propane (R290) and ethane (R170). Based on the present thermodynamic model, the performance optimization on the R290‐R170 cascade refrigerator was made with considerations of degree of subcooling, degree of superheat, evaporation temperature, condensation temperature, cascade condensation temperature, and cascade temperature difference. Variations of the coefficient of performance, exergy destruction, and total exergy efficiency of the refrigeration cycle were analyzed. Two mathematical correlations yielding the optimal cascade condensation temperature and maximized coefficient of performance were developed by multilinear regression analysis. When the evaporation temperature is − 60°C, the maximized coefficient of performance and total exergy efficiency are 1.276 and 49.51%. This paper demonstrates the potential for improving the R290‐R170 cascade refrigeration system and furnishes the basis for further exploration on ultra‐low‐temperature refrigerators.