Background: Ferula vesceritensis is an indigenous plant of Algerian Sahara riche in sesquiterpene coumarins. Objective: In this study, we investigated the biological activity of sesquiterpene coumarins; coladin, ferulenol, and lapiferin (10_-acetoxy-6_-angeloyloxy-8_,9_-epoxy-trans-caxotan-4_-ol) isolated for the first time from the crude extract CH2Cl2—MeOH (1:1) of the roots of F. vesceritensis Coss. et Dur. Materials and Methods: Structures of coladin, ferulenol, and lapiferin were determined by extensive nuclear magnetic resonance (NMR) analyses, including 1D-(1H and13C) and 2D-NMR experiments (correlation spectroscopy, heteronuclear single-quantum coherence, heteronuclear multiple bond correlation (HMBC), and nuclear overhauser effect spectroscopy) as well as high-resolution electron ionization mass spectra and mass spectroscopy analyses. Results: Tested on mouse B16F1 melanoma cells, ferulenol, coladin, and lapiferin exhibited a significant decrease in cell proliferation and a decrease of mitochondrial dehydrogenase activity evaluated on living FAO cells and B16F1 melanoma cells with the WST-1 throughout an apoptotic pathway. They displayed pro-apoptotic effects observed by a decrease in mitochondrial membrane potential and the mitochondrial respiratory rate on isolated liver mitochondria in a dose-dependent manner. Conclusion: Our results highlight the importance of the sesquiterpene coumarins extracted from F. vesceritensis as new biologically active natural products against cancer cells.