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On The Biomedical Promise of Cell Penetrating Peptides: Limits Versus Prospects

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ABSTRACT

The cell membrane poses a substantial hurdle to the use of pharmacologically active biomacromolecules that are not per se actively translocated into cells. An appealing approach to deliver such molecules involves tethering or complexing them with so‐called cell penetrating peptides (CPPs) that are able to cross the plasma membrane of mammalian cells. The CPP approach is currently a major avenue in engineering delivery systems that are hoped to mediate the non‐invasive import of problematic cargos into cells. The large number of different cargo molecules that have been efficiently delivered by CPPs ranges from small molecules to proteins and even liposomes and particles. With respect to the involved mechanism(s) there is increasing evidence for endocytosis as a major route of entry. Moreover, in terms of intracellular trafficking, current data argues for the transport to acidic early endosomal compartments with cytosolic release mediated via retrograde delivery through the Golgi apparatus and the endoplasmic reticulum. The focus of this review is to revisit the performance of cell penetrating peptides for drug delivery. To this aim we cover both accomplishments and failures and report on new prospects of the CPP approach. Besides a selection of successful case histories of CPPs we also review the limitations of CPP mediated translocation. In particular, we comment on the impact of (i) metabolic degradation, (ii) the cell line and cellular differentiation state dependent uptake of CPPs, as well as (iii) the regulation of their endocytic traffic by Rho‐family GTPases. Further on, we aim at the identification of promising niches for CPP application in drug delivery. In this context, as inspired by current literature, we focus on three principal areas: (i) the delivery of antineoplastic agents, (ii) the delivery of CPPs as antimicrobials, and (iii) the potential of CPPs to target inflammatory tissues. © 2007 Wiley‐Liss, Inc. and the American Pharmacists Association J Pharm Sci 97:144–162, 2008

Section snippets

INTRODUCTION

The full therapeutic potential of peptide‐, protein‐, and nucleic acid‐based drugs is frequently compromised by their limited ability to cross the plasma membrane of mammalian cells, resulting in poor cellular access and inadequate therapeutic efficacy.1, 2 Today this hurdle represents a major challenge for the biomedical development and commercial success of many biopharmaceuticals. Over the past decade, however, attractive prospects for a substantial improvement in the cellular delivery of

Successful Delivery by the CPP Approach

The following section covers selected examples of successful drug (or model drug) delivery by the CPP approach. In particular, we focus on distinct in vivo studies, whereas a broader outline is given in the review of Dietz and Bdeltahr.8 A pioneering piece of work on CPP mediated cellular delivery of heterologous proteins was authored by Fawell et al.53 The authors covalently linked Tat peptides to β‐galactosidase, horseradish peroxidase, RNase A, and domain III of Pseudomonas exotoxin A.

Metabolic Degradation

A major obstacle to CPP mediated drug delivery is thought to consist in the often rapid metabolic clearance of the peptides when in contact or passing the enzymatic barriers of epithelia and endothelia. Koppelhus et al.,103 for example, reported that the observed poor intracellular uptake of CPPs might results from quick degradation of the fluorescence labeled CPPs in the cells. Until today, however, despite its general relevance, information on the momentous subject of enzymatic stability and

NEW PERSPECTIVES FOR APPLICATION OF CPPs IN DRUG DELIVERY

As a consequence of the described limitations of CPP mediated translocation, for example, the dependence on the state of cellular differentiation, we aim in this final section at the identification of promising niches for CPP application in drug delivery. Ideally, these approaches should either overcome or bypass the given limitations of CPP mediated drug delivery described above and, thus, offer a realistic avenue toward the usage of CPPs in a therapeutic context. As inspired by current

CONCLUSION

Over more than a decade, a broad variety of CPPs has been evaluated for their capacity to support the cellular delivery of therapeutics that normally do not cross the plasma membrane. Although a number of landmark studies in the field claimed a practically unrestricted cellular access of CPPs and CPP associated cargos, crucial limitations to these shuttles have been pointed out more recently. In this review, we attempted to review distinct aspects of CPP mediated cellular delivery. Besides the

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    The authors are pleased to dedicate this review to Professor William I. Higuchi PhD, Salt Lake City, in recognition of his pioneering contributions to fundamental pharmaceutical research.

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