Triple Enzyme‐Regulated Molecular Hydrogels for Carrier‐Free Delivery of Lonidamine

Abstract Cancerous cells exhibit overexpression of multiple enzymes in various cellular compartments. These enzymes are often undruggable, yet display unique advantages in regulating intracellular self‐assembly and dis‐assembly of small molecules for cancer targeting. Herein, a self‐assembling molecule (LND‐1p‐ES) for carrier‐free delivery of lonidamine specifically to cancerous cells is designed, where LND‐1p‐ES executes as both a drug and a carrier with combined benefits thereof. Under the precise regulation of phosphatase (ALP) and esterase (CES), LND‐1p‐ES is capable of self‐assembling intracellularly in a spatiotemporal manner, to confer lonidamine‐borne nanofibers with enhanced cellular uptake. These nanofibers also facilitate controlled drug release with the aid of cellular proteases. Taking advantages of overexpressing ALP and CES as well as proteases in cancerous cells, the LND‐1p‐ES formulation demonstrates enhanced potency and selectivity against melanoma cells A375 in vitro and in vivo. In comparison, none/single enzyme responsive compounds fail to show a similar potency or selectivity, further confirming the indispensable roles of these enzymes in the delivery system. Collectively, the research provides a viable strategy to utilize multiple enzymes in cancerous cells for regulation of intracellular self‐assembly, which can be expanded to design smart soft materials responsive to multiple biologically relevant biomolecules for enhanced therapeutic efficacy.