This study aims to investigate the chemical constituents and anti-melanogenic activity of Elaeagnus angustifolia L. leaves. The compounds were separated and purified from alcohol extract of E. angustifolia leaves by a series of chromatographic techniques, including silica gel column chromatography, Sephadex LH-20 column chromatography, semi-preparative high performance liquid chromatography and thin-layer chromatography. The structures of the compounds were identified through spectral data and literature reports, their in vitro mushroom tyrosinase (TYR) inhibitory activity and intracellular melanin synthesis inhibitory activity were detected. The regulation effects of the compounds on the mRNA and protein expression of TYR, TYR-related protein 1 (TRP-l), TRP-2 and microphthalmia-associated transcription factor (MITF) in mouse melanoma cells B16F10 were detected by RT-qPCR and Western blot. The concentration of cyclic adenosine monophosphate (cAMP) in B16F10 cells treated with the compound was determined by ELISA. RT-qPCR and Western blot were applied to evaluate the mRNA and protein expression levels of melanin synthesis signaling pathways proteins including protein kinase B (AKT), phosphorylated AKT (p-AKT), β-catenin and phosphorylated β-catenin (p-β-catenin) in compound-treated B16F10 cells. Three compounds were isolated and identified from the leaves of E. angustifolia, namely naphthisoxazol A (1), isorhamnetin-3-O-sophoroside (2), and isorhamnetin 3-O-β-D-sophoroside-7-O-α-L-rhamnoside (3) . The results showed that all three compounds could inhibit B16F10 intracellular TYR activity and decrease the transcription and protein expression of TYR, TRP-1, TRP-2, and MITF genes in B16F10 cells. Compared with the control group, the three compounds significantly inhibited intracellular cAMP production and subsequently reduced the protein expression of p-AKT and nuclear β-catenin, thereby downregulated the expression of MITF and TYR family proteins (TYR, TRP-1 and TRP-2) leading to reduced intracellular melanin synthesis.