藥學(xué)專業(yè)畢業(yè)論文外文翻譯--傳病媒介設(shè)計(jì)與施藥綜述細(xì)胞滲透性肽二十年從分子機(jī)制到治療（英文）

1、THEMED SECTION: VECTOR DESIGN AND DRUG DELIVERYREVIEWTwenty years of cell-penetrating peptides: from molecular mechanisms to therapeuticsFrederic Heitz, May Catherine Morris and Gilles DivitaCentre de Recherches de Bioch

2、imie Macromoléculaire, UMR 5237, CNRS, UM-1, UM-2, CRBM-Department of Molecular Biophysics and Therapeutics, 1919 Route de Mende, Montpellier, FranceThe recent discovery of new potent therapeutic molecules that do n

3、ot reach the clinic due to poor delivery and low bioavailability have made of delivery a key stone in therapeutic development. Several technologies have been designed to improve cellular uptake of therapeutic molecules,

4、including cell-penetrating peptides (CPPs). CPPs were first discovered based on the potency of several proteins to enter cells. Numerous CPPs have been described so far, which can be grouped into two major classes, the f

5、irst requiring chemical linkage with the drug for cellular internalization and the second involving formation of stable, non-covalent complexes with drugs. Nowadays, CPPs constitute very promising tools for non-invasive

6、cellular import of cargo and have been successfully applied for in vitro and in vivo delivery of therapeutic molecules varying from small chemical molecule, nucleic acids, proteins, peptides, liposomes and particles. Thi

7、s review will focus on the structure/function and cellular uptake mechanism of CPPs in the general context of drug delivery. We will also highlight the application of peptide carriers for the delivery of therapeutic mole

8、cules and provide an update of their clinical evaluation. British Journal of Pharmacology (2009) 157, 195–206; doi:10.1111/j.1476-5381.2008.00057.x; published online 20 March 2009This article is part of a themed section

9、on Vector Design and Drug Delivery. For a list of all articles in this section see the end of this paper, or visit: http://www3.interscience.wiley.com/journal/121548564/issueyear?year=2009Keywords: cell-penetrating pepti

10、de; non-covalent delivery system; siRNA; nanoparticle; drug delivery; molecular mechanisms; therapeuticsAbbreviations: CPP, cell-penetrating peptide; GAG, GlucosAminoGlycan; NLS, nuclear localization sequence; PMO, phos-

11、 phorodiamidate morpholino-oligomer; PNA, peptide-nucleic acid; PTD, protein transduction domainsIntroduction: challenges in drug deliveryOver the past 10 years, in order to circumvent limitations ofsmall molecule- and g

12、ene-based therapies, we have witnesseda dramatic acceleration in the production of new large thera-peutic molecules, which do not follow Lipinski’s rules, such asproteins, peptides and nucleic acid therapeutics. However,

13、their development is restricted by very specific issues includ-ing poor stability in vivo, lack of cellular uptake and insuffi-cient capability to reach targets. This is associated with thecomplete loss of pharmaceutical

14、 potency or at least with therequirement for high doses and risk of major side effects.Therefore, delivery constitutes a major piece of the therapeu-tic puzzle, and there is a real demand for new and moreefficient drug d

15、elivery systems. Major rules have to be satis-fied, in particular: (i) delivery efficiency in different and chal-lenging cell lines; (ii) rapid endosomal release; (iii) ability toreach the target; (iv) activity at low do

16、ses; (v) lack of toxicity;and (vi) facility of therapeutic application.Substantial progress has been made in the design of newtechnologies to improve cellular uptake of therapeutic com-pounds (Opalinska and Gewirtz, 2002

17、; Järver and Langel, 2004;Glover et al., 2005; Torchilin, 2005; De Fougerolles et al., 2007;Kong and Mooney, 2007). A number of non-viral strategieshave been proposed including lipid, polycationic, nanopar-ticle and

18、 peptide-based formulations as reported in this specialissue (Morris et al., 2000; Ogris and Wagner, 2002; Järver andLangel, 2004; Torchilin, 2005), but only a subset of thesetechnologies are efficiently applied in

19、vivo at either preclinicalor clinical levels. Protein transduction domains (PTDs) orcell-penetrating peptides (CPPs) correspond to short 30residue synthetic peptides and are part of the most promisingstrategy to overcome

20、 both extracellular and intracellular limi-tations of various biomolecules of including plasmid DNA,Correspondence: Gilles Divita, Centre de Recherches de Biochimie Macro- moléculaire, UMR 5237, CNRS, UM-1, UM-2, CR

21、BM-Department of Molecular Biophysics and Therapeutics, 1919 Route de Mende, 34293 Montpellier, France. E-mail: gilles.divita@crbm.cnrs.fr Received 14 July 2008; revised 7 October 2008; accepted 20 October 2008British Jo

22、urnal of Pharmacology (2009), 157, 195–206 © 2009 The Authors Journal compilation © 2009 The British Pharmacological Society All rights reserved 0007-1188/09www.brjpharmacol.orgCovalent strategyCell-penetrating

23、 peptide-based technologies described so farmainly involve the formation of a covalent conjugatebetween the cargo and the carrier peptide, which is achievedby chemical cross-linking or by cloning followed by expres-sion

24、of a CPP fusion protein (Nagahara et al., 1998; Gait,2003; Moulton and Moulton, 2004; Zatsepin et al., 2005).Most of the work has been reported for peptides derived fromTat (Fawell et al., 1994; Vives et al., 1997; Frank

25、el and Pabo,1988), penetratin (Derossi et al., 1994), polyarginine peptideArg8 sequence (Wender et al., 2000; Futaki et al., 2001) andTransportan, (Pooga et al., 1998). Other protein-derived pep-tides such as VP22 protei

26、n from Herpes Simplex Virus (Elliottand O’Hare, 1997), pVec (Elmquist et al., 2001), calcitonin-derived peptides (Schmidt et al., 1998; Krauss et al., 2004),antimicrobial peptides Buforin I and SynB (Park et al., 1998;Pa

27、rk et al., 2000), as well as polyproline sweet arrow peptide(Pujals et al., 2006) have also been successfully used toimprove the delivery of covalently linked cargos (Joliot andProchiantz, 2004; El-Andaloussi et al., 200

28、5; Murriel andDowdy, 2006). More recently, new generations of CPPs,combining different transduction motifs (Abes et al., 2007)or transduction domains in tandem with protein oroligonucleotide-binding domains (Meade and Do

29、wdy, 2007)have been proposed. Different chemistries have been pro-posed for stable or cleavable conjugation involving mainlydisulfide or thio-esters linkages. According to the stability andefficiency of the cargo, severa

30、l parameters need to be consid-ered including the type of linkage chemistry, the nature of thespacer (Gait, 2003; Zatsepin et al., 2005). Covalent strategieshave been mainly reported for the delivery of DNA mimicmolecule

31、s or steric block oligonucleotides, including PNA(Koppelhus et al., 2002; Fabani et al., 2008), phosphorodiami-date morpholino-oligomer (PMO) (Abes et al., 2006; Lebleuet al., 2008; Moulton and Moulton, 2008), peptide an

32、dprotein (Snyder and Dowdy, 2005). Conjugation methodsoffer several advantages for in vivo applications includingrationalization, reproducibility of the procedure, togetherwith the control of the stoechiometry of the CPP

33、-cargo.However, the covalent CPP technology is limited from achemical point of view and risks altering the biological activ-ity of the cargo. This is particularly true, in the case of chargedoligonucleotide or siRNA, for

34、 which CPP coupling has led torestricted biological activities (Juliano et al., 2008), non-covalent strategies thereby appearing more appropriate.Non-covalent strategyThis strategy is mainly based on short amphipathic pe

35、ptidecarriers consisting of two domains: a hydrophilic (polar)domain and a hydrophobic (non-polar) domain (Table 1). Theamphipathic character may arise from either the primarystructure or the secondary structure. Primary

36、 amphipathicpeptides can be defined as the sequential assembly of adomain of hydrophobic residues with a domain of hydro-philic residues. Secondary amphipathic peptides are gener-ated by the conformational state that all

37、ows positioning ofhydrophobic and hydrophilic residues on opposite sides ofthe molecule (Deshayes et al., 2005). Several CPPs have beenreported to form non-covalent complexes with biomoleculesand to improve their deliver

38、y into mammalian cells (Morriset al., 2008).Non-covalent approach was originally developed for genedelivery; several peptides able to condense DNA associatedwith peptides that favour endosomal escape including fusionpept

39、ide of HA2 subunit of influenza hemaglutinin have beendescribed (Lear and Degrado, 1987; Parente et al., 1990). Syn-thetic peptides analogs GALA, KALA, JTS1 (Gottschalk et al.,1996; Wyman et al., 1997), PPTG1 (Rittner et

40、 al., 2002), MPG(Morris et al., 1999) and histidine-rich peptides (Midoux et al.,1998; Kichler et al., 2003) were also reported as potent genedelivery systems. In 2001, we demonstrated that the amphi-pathic peptide Pep-1

41、 could be successfully applied to thedelivery of small peptides and proteins in a non-covalentapproach (Morris et al., 2001). In 2003, a non-covalent strat-egy based on MPG was shown to efficiently deliver siRNA intocult

42、ured cell lines (Simeoni et al., 2003). Pep-1 and MPG areprimary amphipathic peptides containing a hydrophiliclysine-rich domain derived from the nuclear localizationsequence (NLS) of SV40 large T antigen (KKKRKV), and a

43、variable N-terminal hydrophobic moiety derived form thefusion sequence of the HIV protein gp41 (GALFLGFLGAAG-STMGA) for MPG, and from a tryptophan-rich cluster (KETW-WETWWTEW) for Pep-1, separated by a linker domain, whi

44、chimproves the flexibility and the integrity of both the hydro-phobic and hydrophilic domains (Morris et al., 1997; Morriset al., 1999; Simeoni et al., 2003). MPG and Pep-1 form stablecomplexes with their respective carg

45、o (oligonucleotide orprotein/peptide) through non-covalent electrostatic andhydrophobic interactions (Morris et al., 1997; 1999; 2001;Simeoni et al., 2003; Gros et al., 2006; Munoz-Morris et al.,2007). Non-covalent strat

46、egies for protein and oligonucle-otide delivery have been recently been extended to otherCPPs, including Tat (Meade and Dowdy, 2007), polyarginine(Kim et al., 2006; Kumar et al., 2007) and Transportan-derivedpeptides (Po

47、oga et al., 2001; Lundberg et al., 2007).Cellular uptake mechanism of cell-penetrating peptidesThe cellular uptake mechanism of CPPs is an essential piece ofthe puzzle for the development and optimization of appropri-ate

48、 strategies for in vivo therapeutic applications. Althoughcellular internalization of CPPs was reported in a wide varietyof cell types, their mechanism of internalization remained‘mysterious’ for a long time, as being in

49、dependent of endocy-tosis, of energy and of specific receptor. In the last 5 years, theCPP field has suffered and learnt from technical artifacts. Assuch, in 2003, Lebleu and colleagues, proposed a revised cel-lular upta

50、ke mechanism for CPPs, essentially associated withthe endosomal pathway (Richard et al., 2003). Ever since, themechanism of many CPPs has re-examined and reported to bemediated by endocytosis (Lundberg and Johansson, 200

51、1;Nakase et al., 2004; Wadia et al., 2004; Fischer et al., 2005;Richard et al., 2005; Murriel and Dowdy, 2006). However, formost CPPs, the cellular uptake mechanism still needs to beconfirmed and remains controversial, p