The titratable, double-chain amphiphiles 1,2-dipalmitoyl-sn-3-succinylglycerol (1,2-DPSG), 1,2-dioleoyl-sn-3-succinylglycerol (1,2-DOSG) and 1,3-dipalmitoylsuccinylglycerol (1,3-DPSG) have been used in combination with phosphatidylethanolamine (PE) to form pH-sensitive liposomes. The effect of the compounds on dielaidoyl PE bilayer stabilization was examined by differential scanning calorimetry. Only 1,2-DPSG showed bilayer stabilization activity; whereas the other two are destabilizers at pH 7.4. All three amphiphiles became strong destabilizers at pH 5.0. The ability of the amphiphiles to stabilize DOPE liposomes was examined by light scattering and calcein entrapment. In general, 1,2-DPSG is the most potent stabilizer of PE bilayers while 1,3-DPSG is the weakest liposome stabilizer. All three compounds can be combined with DOPE to generate liposomes which are stable at neutral and basic pH. At weakly acidic pH, the liposomes are leaky and exhibit extensive lipid mixing, with protons and calcium showing synergistic effects on lipid mixing. DOPE/1,2-DPSG liposomes are stable in human plasma and remain acid-sensitive even after prolonged plasma incubation. Immunoliposomes prepared from either DOPE/1,2-DPSG or DOPE/1,2-DOSG can deliver diphtheria toxin A fragment to the cytoplasm of cultured cells in a process which involves endocytosis of the liposomes. Immunoliposomes prepared with 1,2-DPSG are more effective drug carriers than those prepared with 1,2-DOSG. These results indicate that the bilayer- and, hence the liposome-stabilization activity of the diacylsuccinylglycerol depends on the structure of the compounds. The potential drug delivery activity of the pH-sensitive liposomes composed of these lipids is discussed.