Direct cytosolic delivery of cargoes in vivo by a chimera consisting of D- and L-arginine residues

• Published in: Journal of controlled release Vol. 162; no. 2; pp. 286 - 294
• Main Authors: Ma, Yan, Gong, Cheng, Ma, Yilong, Fan, Fengkai, Luo, Meijie, Yang, Fei, Zhang, Yu-Hui
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier B.V 10.09.2012; Elsevier
• Subjects: Animals; Arginine - chemistry; bioavailability; Biological and medical sciences; Biological Transport; Cell Membrane - metabolism; Cell Survival - drug effects; Cell-penetrating peptide; Cell-Penetrating Peptides - administration & dosage; Cell-Penetrating Peptides - chemistry; Cell-Penetrating Peptides - pharmacokinetics; chimerism; cultured cells; cytosol; Cytosol - metabolism; Cytosolic delivery; Drug Carriers - administration & dosage; Drug Carriers - chemistry; Drug Carriers - pharmacokinetics; drugs; endocytosis; Fluorescent Dyes - administration & dosage; Fluorescent Dyes - pharmacokinetics; General pharmacology; guidelines; HEK293 Cells; HeLa Cells; Heterogeneous; Humans; hybrids; intravenous injection; Medical sciences; membrane potential; Mice; Oligoarginine; Oligopeptides - administration & dosage; Oligopeptides - chemistry; Oligopeptides - pharmacokinetics; peptides; Pharmaceutical technology. Pharmaceutical industry; Pharmacology. Drug treatments; Rhodamines - administration & dosage; Rhodamines - pharmacokinetics; Systemic toxicity; therapeutics; toxicity; Heterogeneous; Oligoarginine; Systemic toxicity; Cytosolic delivery; Cell-penetrating peptide; Pharmaceutical technology; Cell penetrating peptide; Toxicity; Cytosol; In vivo; Arginine; Residue; Aminoacid; Chimera
• ISSN: 0168-3659; 1873-4995; 1873-4995
• DOI: 10.1016/j.jconrel.2012.07.022

The ability of cell-penetrating peptides (CPPs) to deliver a range of membrane-impermeable molecules into living cells makes them attractive potential vehicles for therapeutics. However, in vivo, the efficiency of CPP delivery to the cytosol remains unsatisfactory owing to endosomal entrapment and/or systemic toxicity, which severely restrict their bioavailability and efficacy in in vivo applications. In this study, we developed a series of novel chimeras consisting of various numbers of d- and l-arginine residues and investigated their cellular uptake behaviors and systemic toxicities. We demonstrated that the intracellular distribution, uptake efficiency, and systemic toxicity of these oligoarginines were all significantly affected by the number of d-arginine residues in the peptide sequence. We also found that a hybrid peptide, (rR)3R2, possessed low systemic toxicity, high uptake efficiency, and, remarkably, achieved efficient cytosolic delivery not only in cultured cells but also in living tissue cells in mice after intravenous injection, implying that this heterogeneous motif might have promising applications in the delivery of cargoes of small sizes directed to cytosolic targets in vivo. Our studies into the uptake mechanism of (rR)3R2 indicate that its cellular uptake was not affected by pharmacological or physical inhibitors of endocytosis but by the elimination of the membrane potential, suggesting that (rR)3R2 does not enter the cells via endocytosis but rather through direct membrane translocation driven by the membrane potential. The results here might provide useful guidelines for the design and application of CPPs in drug delivery. [Display omitted]