In this study, we developed redox‐sensitive vesicles using synthesised lipoyl cholesterol derivatives, a non‐ionic surfactant and an optimum level of free cholesterol. Interestingly, concentration‐dependent self‐assembly was observed by scanning electron microscopy, wherein vesicles manifested as hollow spherical (at 0.15 mm) and triangular (0.50 mm). The redoxresponsive characteristics of the vesicles was probed in the presence of dithiothreitol; they underwent a clear increase in size as observed by dynamic light scattering measurements. These vesicles could easily encapsulate an anticancer drug, doxorubicin, and were observed to be stable in the presence of serum. They showed substantial release of the drug in response to biologically relevant stimulus, that is, glutathione. A toxicity assessment on HeLa and HepG2 cancer cells demonstrated activities of the drug‐loaded vesicles comparable to that of free drug, whereas significantly enhanced toxicity and apoptotic induction were observed against drug‐resistant HeLa cells, which was determined by studying the cellular internalisation of doxorubicin.