Diabetic patients have a higher risk of health problems, including heart attack, stroke, kidney failure, and permanent vision loss. Ehretia anacua is a plant used in folk medicine for the management of diabetes. The current study investigated the phytochemical profile of Ehretia anacua and evaluated its antioxidant activity, acute toxicity, and therapeutic potential against streptozotocin-induced diabetes in a rat model. Additionally, we evaluated the phenolic and flavonoid content either colorimetrically or by HPLC analysis. Eleven compounds were isolated namely, phytyl palmitate (1), β-sitosterol tetracosanoate (2), palmitic acid (3), isophytol (4), β-sitosterol (5), β-sitosterol-3-O-β-D-glucopyranoside (6), methyl rosmarinate (7), caffeic acid (8), (6R,7E,9R)-9-hydroxy-megastigma-4,7-dien-3-one 9-O-D-glucopyranoside (9), roseoside (10) and rosmarinic acid (11). These results revealed that E. anacua is a rich source of phenolic compounds. The ethyl acetate and n-butanol fractions have potent antioxidant activity with value of IC50 is 25.16 ± 0.09 and 87.84 ± 0.39 µg/mL, respectively. Moreover, the ethyl acetate fraction exhibited superior antidiabetic efficacy by significantly reducing blood sugar levels and lipid profiles and modulating related biochemical parameters. These results suggest that E. anacua leaves could serve as a potential source of antidiabetic and antioxidant medications, possibly offering new avenues for combination therapy for diabetes treatment.