Melittin is a 26-residue, amphipathic, cell-penetrating, α-helical anti-hepatoma peptide isolated from bee venom. However, the application of melittin as a drug is limited owing to its original conformational flexibility and low stability. In this study, we designed, synthesized, and tested a series of hydrocarbon-stapled analogs of melittin, of which, some analogs showed remarkable enhancement not only in anti-hepatoma activity, but also in α-helicity and protease resistance when compared to the parent melittin. These results disclosed the important impact of all-hydrocarbon crosslinking on the biological activity, stability, and hemolytic activity of melittin.