Charge-guided masking of a membrane-destabilizing peptide enables efficient endosomal escape for targeted intracellular delivery of proteins

• DOI: 10.1016/j.apsb.2024.06.022
• Publication Date: 2024-06-26T16:14:56Z
• Authors: Yan Zhao; Haolin Jiang; Hang Chen; Jiazhen Yu; Luyao Wang; Wen Zhou; Juanjuan Du
• Keywords: pH responsiveness; LMWP; Cell-penetrating peptides; Original Article; Therapeutics. Pharmacology; RM1-950; Endosomal escape; Cytosolic delivery; Antibody

Intracellular delivery of biologicals such as peptides, proteins, and nucleic acids presents a great opportunity for innovative therapeutics. However, the endosome entrapment remains major bottleneck in intracellular biomacromolecules, largely limiting their therapeutic potential. Here, we converted cell-penetrating peptide (CPP), low molecular weight protamine (LMWP), to endosomal escape peptides (EEPs) by masking LMWP with pH-responsive counter-ionic peptide. The resulting masked CPPs (mLMWP mLMWP2) effectively promoted peptide/protein cargoes from endosomes into cytoplasm. Consequential lysosome repair lysophagy were initiated upon endolysosomal leakage. Minimal reactive oxygen species (ROS) elevation or cell death was observed. Based on mLMWP2, constructed an protein system containing antibody targeting module, mLMWP2 desired cargo. With HER2-targeting system, efficiently translocated cyclization recombination enzyme (Cre) BH3-interacting domain agonist (BID) cytosol HER2