Stimuli-Responsive Polypeptide Nanoparticles for Enhanced DNA Delivery

• Published in: Molecules (Basel, Switzerland) Switzerland), 2022-12, Vol.27 (23), p.8495
• Main Authors: Korovkina, Olga, Polyakov, Dmitry, Korzhikov-Vlakh, Viktor, Korzhikova-Vlakh, Evgenia
• Format: Article
• Language: English
• Subjects: Amino Acids; Copolymerization; Copolymers; cross-linked nanoparticles; Cysteine; Cytoplasm; Deoxyribonucleic acid; Disulfide bonds; Disulfides; DNA; DNA - chemistry; Drug Delivery Systems; Enzymes; gene delivery; Gene therapy; Genes; Glutamate; Glutamic acid; HEK293 Cells; Histidine; Humans; Hydrogen-Ion Concentration; Lysine; Nanoparticles; Nanoparticles - chemistry; Nucleic acids; Peptides - chemistry; pH and redox-responsive delivery systems; Phenylalanine; Physiology; Polymerization; Polymers; Polypeptides; Self-assembly; Surface charge; System effectiveness; Transfection
• ISSN: 1420-3049; 1420-3049
• DOI: 10.3390/molecules27238495

The development of non-viral delivery systems for effective gene therapy is one of the current challenges in modern biomedicinal chemistry. In this paper, the synthesis of pH- and redox-responsive amphiphilic polypeptides for intracellular DNA delivery is reported and discussed. Two series of polypeptides consisting of L-lysine, L-phenylalanine, L-histidine, and L-cysteine as well as the same amino acids with L-glutamic acid were synthesized by a combination of copolymerization of N-carboxyanhydrides of α-amino acids and post-polymerization modification of the resulting copolymers. The presence of histidine provided pH-sensitive properties under weakly acidic conditions specific to endosomal pH. In turn, the presence of cysteine allowed for the formation of redox-responsive disulfide bonds, which stabilized the self-assembled nanoparticles in the extracellular environment but could degrade inside the cell. The formation of intraparticle disulfide bonds resulted in their compactization from 200-250 to 55-100 nm. Empty and pDNA-loaded cross-linked nanoparticles showed enhanced stability in various media compared to non-crosslinked nanoparticles. At the same time, the addition of glutathione promoted particle degradation and nucleic acid release. The delivery systems were able to retain their size and surface charge at polypeptide/pDNA ratios of 10 or higher. GFP expression in HEK 293 was induced by the delivery of pEGFP-N3 with the developed polypeptide nanoparticles. The maximal transfection efficacy (70%) was observed when the polypeptide/pDNA ratio was 100.