Enantiomeric and Diastereoisomeric (Mixed) L/ D-Octaarginine Derivatives - A Simple Way of Modulating the Properties of Cell-Penetrating Peptides

• Published in: Chemistry & biodiversity 2013-07, Vol.10 (7), p.1165-1184
• Main Authors: Purkayastha, Nirupam, Eyer, Klaus, Robinson, Tom, Dittrich, Petra S., Beck, Albert K., Seebach, Dieter, Kolesinska, Beata, Cadalbert, Riccardo
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Amino acids; Cell Membrane Permeability; Cell penetration; Cell-Penetrating Peptides - blood; Cell-Penetrating Peptides - chemistry; Cell-Penetrating Peptides - metabolism; Fluorescein - chemistry; Half-Life; HEK Cells; HEK293 Cells; Humans; Lipid Bilayers - chemistry; Microfluidic devices; Microscopy, Confocal; Oligoarginines; Oligopeptides - blood; Oligopeptides - chemistry; Oligopeptides - metabolism; Peptides; Phospholipid vesicles; Proteins; Stereoisomerism
• ISSN: 1612-1872; 1612-1880
• DOI: 10.1002/cbdv.201300180

Cell‐penetrating peptides (CPPs) are promising vehicles for delivery of drugs, antibiotics, proteins, nucleic acid derivatives, etc. into eukaryotic and prokaryotic target cells. To prevent premature degradation, CPPs consisting of D‐ or β‐amino acid residues have been used. We present simple models for the various modes of delivery of physiologically active cargoes by CPPs, depending on the nature of their conjugation (Fig. 1), and we describe the plasma stability of oligoarginines (OAs) 1–4, the most common unnatural CPPs. Fluorescein‐labeled L‐octaarginine 1 was found to have a half‐life (t1/2) of 7 d (Fig. 2). For possible medicinal applications, the former type of derivative would be too unstable, and the latter one undesirably persistent. Thus, seven of the 256 possible ‘mixed’ Flua‐L/D‐octaarginine amides, 4a–4g, were synthesized and shown to have half‐lives in heparine‐stabilized human plasma between 8 min and 5.5 h (Figs. 3 and 4). The cell penetration of the new OAs was investigated with ‘healthy’ and with apoptotic HEK cells (Figs. 5–8), and their interactions with phospholipid bilayers were studied, using anionic lipid vesicles (Figs. 9 and 10). There are surprisingly large differences in the rates of cell penetration and binding to vesicle walls between the various stereoisomeric octaarginine derivatives 1, 2, and 4a–4g (Figs. 5 and 7). – The role of D‐amino acids and D‐peptides in nature and in drug design is briefly discussed and referenced.