This study aimed to investigate the effect of aqueous ethanol extract of Cuminum cyminum (AEECC) on oxidative stress, inflammation and overactivity of the urinary bladder induced by cyclophosphamide (CYP). The enhanced nociception behavior, bladder weight, vascular permeability, edema, hemorrhage, nitric oxide, IL‐6 and TNF‐α levels by CYP administration were significantly reduced by AEECC (250 and 500 mg/kg). A significant increase in serum antioxidant systems such as CAT and GPx was also observed in AEECC‐treated rats. The AEECC (3 mg/ml) significantly reduced urinary bladder tone in the strips pre‐contracted with carbachol in both control and CYP‐treated rats. This relaxation was demolished by atropine, nifedipine, glibenclamide, and indomethacin but not with propranolol. The plant extract showed the presence of antioxidant and anti‐inflammatory phytochemicals. These results suggest that Cuminum cyminum offers uroprotective activity and can ameliorate CYP‐induced bladder toxicity by modulating antioxidant parameters, pro‐inflammatory cytokine levels and bladder smooth muscle overactivity. The in silico binding interactions of anti‐oxidant 2I3Y and anti‐inflammatory protein 1TNF with various ligands from Cuminum cyminum seeds reveal potential bioactive compounds with promising anti‐oxidant and anti‐inflammatory properties, providing valuable insights for drug development and nutraceutical research