Drought affects crop developmentnand growth. To explore the physiological effects of drought stress on soybean, HeiNong44 (HN44) and HeiNong65 (HN65) varieties were used as experimental materials and PEG-6000 was used as the osmotic medium. The antioxidant enzyme activity, osmotic adjustment substance content, antioxidant capacity, and endogenous hormone content of the two soybean varieties were studied under different drought degrees and different treatment durations. Drought stress caused significant physiological changes in soybean. The antioxidant enzyme activities, osmoregulation substance content, and total antioxidant capacity (T-AOC) of HN65 and HN44 showed an increasing trend under mild and moderate drought, however, they first increased and then decreased under severe drought conditions. Following the extension of treatment time, malondialdehyde (MDA) showed an increasing trend. As drought increased, gibberellin (GA) content showed a decreasing trend, while abscisic acid (ABA), salicylic acid (SA), and zeatin nucleoside (ZA) content showed an increasing trend. The auxin (IAA) content of the two varieties showed opposite change trends. In short, drought had a significant impact on the physiology of these two soybean varieties; however, overall, the drought resistance of HN65 was lower than that of HN44. This study provides a research theoretical basis for addressing the drought resistance mechanism and the breeding of drought resistant soybean varieties.