BACKGROUND: Coumarins are naturally occurring biologically active heterocyclic molecules endowed with a wide range of biological properties, including antibacterial, antifungal and antitumor activities. OBJECTIVE: The present work aimed to synthesize new coumarin-containing compounds and to investigate their cytotoxic activity. METHODS: Coumarin peptide and coumarin amino alcohols were prepared by treating epoxide-containing coumarin derivatives with suitable aromatic amines and peptides in trifluoroethanol as a solvent at 50 °C. These derivatives were evaluated for their cytotoxic activity on three different cell lines: HeLa, MDA-MB-231 and L-132. Cell viability was determined by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) based assay. RESULTS: A new protocol has been developed for the synthesis of thirteen novel coumarin peptide and coumarin amino alcohol derivatives. Among the tested compounds, three derivatives showed significant activity against all the tested cell lines. Docking studies indicated favorable interactions of the di-substituted peptide coumarin derivatives with the Asp 351 and Thr 347 amino acids at the active site of human estrogen receptor. CONCLUSIONS: The results suggest that the newly synthesized compounds may be promising candidates in the research of new antitumor compounds.