Enzymatically Active Liposomal Nanobots for the Penetration of Intercellular and Intracellular Barriers

ABSTRACT

Delivery systems for nanomedicine are critical, however, they are restricted by intercellular and intracellular barriers including lysosomal sequestration after cellular uptake, that impede penetration from the tissue to the subcellular level. Here, we report a liposomal nanobot with enzymatic propulsion and drug‐payload capabilities that exhibits tumor microenvironment responsiveness and active penetration, enabling intracellular transport and active lysosomal escape. This nanobot arises from phase‐separated vesicles of liposome and bacterial outer membrane vesicles that possess doxorubicin‐encapsulation capability and are asymmetrically functionalized with glucose oxidase to form a Janus catalytic interface for self‐diffusiophoresis in the presence of glucose. Under mildly acidic conditions, these nanobots display enhanced motility and chemotactic migration toward acidic regions, leveraging pH gradients in the tumor microenvironment to infiltrate tumor spheroids. Once internalized, the nanobots are rapidly captured by lysosomes and subsequently escape, as a plausible chemo–mechanical mechanism involving catalytic membrane perturbation and active motion. Then nanobot‐associated signals redistribute toward ER‐ and Golgi‐associated compartments and persist across successive recipient cells, supporting intercellular relay transport. The encapsulated drug is released within tumor cells, enabling enhanced antitumor efficacy. This nanobot thus demonstrates an active drug delivery system capable of traversing cellular barriers, providing a strategy for precision tumor therapy.