An enzyme-activated and structural transformable supramolecular oncolytic peptide for efficient cancer immunotherapy through systemic administration

Oncolytic peptides have emerged as a promising oncolytic therapeutic. However, oncolytic peptides face numerous challenges when administrated systemically, including untargeted toxicity, poor bioactivity, and inadequate stability. We here present an innovative strategy to develop self-assembled supramolecular oncolytic peptides suitable for systemic intravenous administration. D2RP (Nap-G^DF^DF^DY-RR-^DKRFYVVMWK^DK) comprises and significantly endows PKHB1 (^DKRFYVVMWK^DK; an endogenous ligand of CD47) with powerful oncolytic activities by self-assembling into positive-charged and α-helix-enriched membrane-lytic nanofibrils. Subsequently, two phosphorylated precursors of D2RP at ^DY4 and Y10, namely pD2RP (Nap-G^DF^DFp^DY-RR-^DKRFYVVMWK^DK) and D2RpP (Nap-G^DF^DF^DY-RR-^DKRFpYVVMWK^DK), respectively, are tailored to generate membrane-lytic nanofibrils in alkaline phosphatase (ALP)-responsive manner. Both pD2RP and D2RpP are biocompatible, while they preorganize into different states and proceed with varied dephosphorylation processes to generate membrane-lytic nanofibrils differing in supramolecular structures and oncolytic activities. Mechanistically, pD2RP and D2RpP remain inactive until dephosphorylated; at this point, their membrane-lytic activities are activated through structural reconfiguration upon reaching the ALP-enriched tumor lesions. When administrated intravenously, pD2RP outperforms D2RpP in tumor rejection by inducing much more potent immune cell death and evoking robust systemic anti-tumor immune responses. This study provides a new paradigm for tailoring supramolecular oncolytic peptides for systemic intravenous administration and a valuable strategy for optimizing their anti-tumor efficacy.