Document Type

Article

Publication Date

11-29-2018

Abstract

Cyclic peptides containing tryptophan (W) and arginine (R) residues, [WR]5, [WR]6, [WR]7, [WR]8, and [WR]9, were synthesized through Fmoc solid-phase chemistry to compare their molecular transporter efficiency. The in vitro cytotoxicity of the peptides was evaluated using human leukemia carcinoma cell line (CCRF-CEM) and normal kidney cell line (LLC-PK1). [WR]6, [WR]7, [WR]8, and [WR]9 were not significantly cytotoxic to LLC-PK1cells at a concentration of 10 μM after 3 h incubation. Among all the peptides, [WR]9 was found to be a more efficient transporter than [WR]5, [WR]6, [WR]7, and [WR]8 in CCRF-CEM cells for delivery of a cell-impermeable fluorescence-labeled negatively charged phosphopeptide (F′-GpYEEI). [WR]9 (10 μM) improved the cellular uptake of F′-GpYEEI (2 μM) by 20-fold. The cellular uptake of a fluorescent conjugate of [WR]9, F′-[W9R8K], was increased in a concentration- and time-dependent pattern in CCRF-CEM cells. The uptake of F′-[W9R8K] was slightly reduced in CCRF-CEM cells in the presence of different endocytic inhibitors, such as nystatin, 5-(N-ethyl-N-isopropyl)amiloride, chlorpromazine, chloroquine, and methyl β-cyclodextrin. Furthermore, the uptake of F′-[W9R8K] was shown to be temperature-dependent and slightly adenosine 5′-triphosphate-dependent. The intracellular/cellular localization (in the nucleus and cytoplasm) of F′-[W9R8K] was confirmed by fluorescent microscopy in CCRF-CEM cells. These studies suggest that large cyclic peptides containing arginine and tryptophan can be used as a molecular transporter of specific compounds.

Comments

This article was originally published in ACS Omega, volume 3, in 2018. DOI: 10.1021/acsomega.8b02589

This is an open access article published under an ACS AuthorChoice License, which permits copying and redistribution of the article or any adaptations for non-commercial purposes.

Copyright

American Chemical Society

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