Membranes that destroy anticancer peptides can bind to negatively charged cancer cell membranes through electrostatic interactions, destroying their functions and leading to cancer cell necrosis. Temporin-1CEa, obtained from the skin secretions of the Chinese frog Rana chensinensis, is an anticancer peptide with 17 amino acid residues that exhibits concentration-dependent cytotoxicity against a variety of cancer cell lines, although it has no obvious cytotoxicity to normal HUVECs. In this work, we designed and synthesized 12 derivative peptides through double-cysteine scanning of temporin-1CEa-truncated peptides. Most of these peptides had greater anticancer activity than the lead peptide temporin-1CEa. Among these derivative peptides, Nu-7 had the strongest anticancer activity. Nu-7 has a greater α-helicity than does temporin-1CEa. We connected Nu-7 to podophyllotoxin through a reduction-responsive linker to obtain Nu-7-1, which showed better anticancer activity than free podophyllotoxin. Nu-7-1 was less toxic to HUVECs and had low hemolytic activity at therapeutic concentrations (although Nu-7-1 showed hemolytic activity at 100 μM). Nu-7-1 functions through two mechanisms: damage to cell membranes and promotion of cell apoptosis. Nu-7-1 is less toxic to normal HUVECs than is podophyllotoxin and shows better safety. In summary, we carried out a series of modifications on temporin-1CEa, among which the anticancer activity of Nu-7-1 was significantly improved compared with that of the lead peptide temporin-1CEa, providing a useful reference for the structural modification of anticancer peptides.