The review focuses on biomimetic approach to the designing multifunctional nanosystems and their application in modern nanotechnology, particularly in the fields of nanomedicine, enzymology, catalysis, and drug delivery. The throughline integrating the chapters of the review is the family of amphiphilic compounds, including surfactants, macrocycles, and mixed compositions. These are very attractive building blocks due to their ability to cooperative or stoichiometric noncovalent interactions with the formation of smart supramolecular assemblies responsive to external stimulus, demonstrating tunable structural and functional behavior. Special attention is devoted to mixed micellar systems, metallosurfactants and macrocycles of different classes, including cyclodextrins, calixarenes, cucurbiturils, pillararenes. These amphiphilic assemblies are widely used as micellar nanocontainers for drug solubilization, biomimetic nanocatalysts for hydrolysis and oxidation-reduction reactions, and artificial ion channels. Amphiphilic molecules, due to their structural similarity to lipid molecules, can integrate with lipid membranes, modifying their properties. This makes amphiphiles important for developing drug delivery systems that have improved bioavailability, stability, prolonged circulation time, and the ability to overcome biological barriers, ultimately improving the effectiveness of pharmacotherapy.