Spontaneous Formation of Ultrasmall Unilamellar Vesicles in Mixtures of an Amphiphilic Drug and a Phospholipid

[Image: see text] We have observed ultrasmall unilamellar vesicles, with diameters of less than 20 nm, in mixtures of the tricyclic antidepressant drug amitriptyline hydrochloride (AMT) and the unsaturated zwitterionic phospholipid 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) in physiological saline solution. The size and shape of spontaneously formed self-assembled aggregates have been characterized using complementary techniques, i.e., small-angle neutron and X-ray scattering (SANS and SAXS) and cryo-transmission electron microscopy (cryo-TEM). We observe rodlike mixed micelles in more concentrated samples that grow considerably in length upon dilution, and a transition from micelles to vesicles is observed as the concentration approaches the critical micelle concentration of AMT. Unlike the micelles, the spontaneously formed vesicles decrease in size with each step of dilution, and ultrasmall unilamellar vesicles, with diameters as small as about 15 nm, were observed at the lowest concentrations. The spontaneously formed ultrasmall unilamellar vesicles maintain their size for as long we have investigated them (i.e., several months). To the best of our knowledge, such small vesicles have never before been reported to form spontaneously in a biocompatible phospholipid-based system. Most interestingly, the size of the vesicles was observed to be strongly dependent on the chemical structure of the phospholipid, and in mixtures of AMT and the phospholipid 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), the vesicles were observed to be considerably larger in size. The self-assembly behavior in the phospholipid–drug surfactant system in many ways resembles the formation of equilibrium micelles and vesicles in mixed anionic/cationic surfactant systems.