In the current study, we investigated the anticancer potential against human colon cells (Caco-2) of colloidal nanosilver (CN–Ag) produced in Syria using bioactive compounds in the aqueous extract of Eucalyptus camaldulensis leaves (AEECL). The formation of AgNPs was confirmed by UV-visible spectroscopy analysis with surface plasmon peak at 449 nm and their average size was found to be 12, 10, 23 nm by SEM, DLS and NTA respectively. This small size has confirmed the effective role of AEECL as capping agent. Further morphological characterization was done by EDS showed the presence of metallic silver. Zeta potential value (-23 mV) indicated the repulsion among the particles and stability of the formulation nanosilver. The anticancer effect of synthesized CN–Ag against Caco-2 has been tested. The cytotoxicity assay showed a dose-dependent and a time-dependent effect of CN–Ag. The high cytotoxicity of CN–Ag at low concentration (5μ/mL) open new prospects for the development of novel therapeutic approaches against human colon cancer Caco-2.