Dual-modified penetratin peptides: Enhancing nucleic acid delivery through stapling and endosomal escape domain

• Published in: Bioorganic & medicinal chemistry 2024-09, Vol.111, p.117871, Article 117871
• Main Authors: Horikoshi, Kanako, Miyamoto, Maho, Tsuchiya, Keisuke, Yokoo, Hidetomo, Demizu, Yosuke
• Format: Article
• Language: English
• Subjects: Cell-penetrating peptides; Endosomal escape domain; Nucleic acid delivery; pDNA; Penetratin; Stapling structures
• ISSN: 0968-0896; 1464-3391; 1464-3391
• DOI: 10.1016/j.bmc.2024.117871

[Display omitted] •This study aims to enhance pDNA delivery using dual-modified penetratin peptides with stapling structures and endosomal escape domains.•Pep-1 to Pep-6 were synthesized with stapling structures and EEDs at the N- or C-terminus.•Peptides with EEDs (Pep-2 and Pep-3) showed significantly higher intracellular uptake than unmodified penetratin (Pep-1).•Stapled peptides (Pep-4 to Pep-6) achieved higher transfection efficiency compared to non-stapled peptides, with Pep-6 being the most effective. Cell-penetrating peptides (CPPs) are crucial for delivering macromolecules such as nucleic acids into cells. This study investigates the effectiveness of dual-modified penetratin peptides, focusing on the impact of stapling structures and an endosomal escape domain (EED) on enhancing intracellular uptake. Some CPPs were synthesized with an EED at either the N- or C-terminus and stapling structures, and then complexed with plasmid DNA (pDNA) to evaluate their cellular uptake. Results revealed that the combination of stapling and an EED significantly improved delivery efficiency, primarily via macropinocytosis and clathrin-mediated endocytosis. These findings underscore the importance of optimizing CPP sequences for effective nucleic acid delivery systems.