Applications and Challenges for Use of Cell-Penetrating Peptides as Delivery Vectors for Peptide and Protein Cargos

Abstract

The hydrophilic nature of peptides and proteins renders them impermeable to cell membranes. Thus, in order to successfully deliver peptide and protein-based therapeutics across the plasma membrane or epithelial and endothelial barriers, a permeation enhancing strategy must be employed. Cell-penetrating peptides (CPPs) constitute a promising tool and have shown applications for peptide and protein delivery into cells as well as across various epithelia and the blood-brain barrier (BBB). CPP-mediated delivery of peptides and proteins may be pursued via covalent conjugation of the CPP to the cargo peptide or protein or via physical complexation obtained by simple bulk-mixing of the CPP with its cargo. Both approaches have their pros and cons, and which is the better choice likely relates to the physicochemical properties of the CPP and its cargo as well as the route of administration, the specific barrier and the target cell. Besides the physical barrier, a metabolic barrier must be taken into consideration when applying peptide-based delivery vectors, such as the CPPs, and stability-enhancing strategies are commonly employed to prolong the CPP half-life. The mechanisms by which CPPs translocate cell membranes are believed to involve both endocytosis and direct translocation, but are still widely investigated and discussed. The fact that multiple factors influence the mechanisms responsible for cellular CPP internalization and the lack of sensitive methods for detection of the CPP, and in some cases the cargo, further complicates the design and conduction of conclusive mechanistic studies.

Keywords: biological barriers; cell-penetrating peptides; delivery vectors; drug delivery; peptide and protein drugs.

Figure 1

Applications for cell-penetrating peptides for the delivery of peptides and proteins into cells and across epithelia and the blood-brain barrier. Dashed arrows specifies target delivery site being in the cytosol or reached by systemic circulation.

Figure 2

Simple illustration of mechanisms by which cell-penetrating peptides have been suggested to traverse the plasma membrane by endocytosis or direct translocation. Dimensions are obtained from [77]. Modified from [96] with permission from Elsevier. Blue helixes represent the CPPs, the arrow the uptake process subsequent to membrane interaction.