Abstract
Naturally occurring cationic antimicrobial peptides exhibit not only antimicrobial activity, but also anticancer activity and are expected to be new weapons in cancer treatment. The selectivity for cancer cells over normal cells is at least partly due to the more negative surface of cancer cells. A lower pH in tumor tissue (pH 6.2–6.9) than that in normal tissues (pH 7.3–7.4) has also been utilized to develop anticancer agents. However, cytotoxicity against normal cells at physiological pH is often an issue. Furthermore, acidic regions can be found in some normal tissues such as the kidneys. Therefore, existing approaches to cancer targeting are not fully satisfactory. In this study, we designed a peptide, HE (GIHHWLHSAHEFGEHFVHHIMNS-amide), with a charge that reverses from −1.5 at pH 7.4 to +6 at pH 5.5 for cancer targeting at low pH based on the antimicrobial peptide magainin 2 by introducing 6 His, an additional Glu, and an amidated terminal. HE interacted with cancer-mimicking negatively charged liposomes in a pH-dependent fashion with a midpoint with a pH of 6.5 just above the membrane surface. The peptide killed human renal adenocarcinoma ACHN cells at pH 6.0, but not at pH 7.4, and was nontoxic against human normal glomerular mesangial cells even at this low pH. Thus, the novel peptide may be a promising lead peptide for cancer therapy, although this derivatization resulted in weakened cytotoxicity.
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References
Al-Benna S, Shai Y, Jacobsen F, Steinstraesser L (2011) Oncolytic activities of host defense peptides. Int J Mol Sci 12:8027–8051
Andreev OA, Dupuy AD, Segala M, Sandugu S, Serra DA, Chichester CO, Engelman DM, Reshetnyak YK (2007) Mechanism and uses of a membrane peptide that targets tumors and other acidic tissues in vivo. Proc Natl Acad Sci USA 104:7893–7898
Apponyi MA, Pukala TL, Brinkworth CS, Maselli VM, Bowie JH, Tyler MJ, Booker GW, Wallace JC, Carver JA, Separovic F, Doyle J, Llewellyn LE (2004) Host-defence peptides of Australian anurans: structure, mechanism of action and evolutionary significance. Peptides 25:1035–1054
Baker MA, Maloy WL, Zasloff M, Jacob LS (1993) Anticancer efficacy of magainin2 and analogue peptides. Cancer Res 53:3052–3057
Binder H, Gawrisch K (2001) Dehydration induces lateral expansion of polyunsaturated 18:0–22:6 phosphatidylcholine in a new lamellar phase. Biophys J 81:969–982
Böhme D, Beck-Sickinger AG (2015) Drug delivery and release systems for targeted tumor therapy. J Pept Sci 21:186–200
Cardone RA, Casavola V, Reshkin SJ (2005) The role of disturbed pH dynamics and the Na+/H+ exchanger in metastasis. Nat Rev Cancer 5:786–795
Cruciani RA, Barker JL, Zasloff M, Chen H-C, Colamonici O (1991) Antibiotic magainins exert cytolytic activity against transformed cell lines through channel formation. Proc Natl Acad Sci USA 88:3792–3796
Dathe M, Wieprecht T, Nikolenko H, Handel L, Maloy WL, MacDonald DL, Beyermann M, Bienert M (1997) Hydrophobicity, hydrophobic moment and angle subtended by charged residues modulate antibacterial and haemolytic activity of amphipathic helical peptides. FEBS Lett 403:208–212
Fjell CD, Hiss JA, Hancock RE, Schneider G (2012) Designing antimicrobial peptides: form follows function. Nat Rev Drug Discov 11:37–51
Haney EF, Hancock RE (2013) Peptide design for antimicrobial and immunomodulatory applications. Biopolymers 100:572–583
Hoskin DW, Ramamoorthy A (2008) Studies on anticancer activities of antimicrobial peptides. Biochim Biophys Acta 1778:357–375
Imura Y, Choda N, Matsuzaki K (2008) Magainin 2 in action: distinct modes of membrane permeabilization in living bacterial and mammalian cells. Biophys J 95:5757–5765
Imura Y, Nishida M, Matsuzaki K (2007) Action mechanism of PEGylated magainin 2 analogue peptide. Biochim Biophys Acta 1768:2578–2585
Iwasaki T, Ishibashi J, Tanaka H, Sato M, Asaoka A, Taylor D, Yamakawa M (2009) Selective cancer cell cytotoxicity of enantiomeric 9-mer peptides derived from beetle defensins depends on negatively charged phosphatidylserine on the cell surface. Peptides 30:660–668
Jin E, Zhang B, Sun X, Zhou Z, Ma X, Sun Q, Tang J, Shen Y, Van Kirk E, Murdoch WJ, Radosz M (2013) Acid-active cell-penetrating peptides for in vivo tumor-targeted drug delivery. J Am Chem Soc 135:933–940
Matsuzaki K (2009) Control of cell selectivity of antimicrobial peptides. Biochim Biophys Acta 1788:1687–1692
Matsuzaki K, Harada M, Funakoshi S, Fujii N, Miyajima K (1991) Physichochemical determinants for the interactions of magainins 1 and 2 with acidic lipid bilayers. Biochim Biophys Acta 1063:162–170
Matsuzaki K, Murase O, Tokuda H, Funakoshi S, Fujii N, Miyajima K (1994) Orientational and aggregational states of magainin 2 in phospholipid bilayers. Biochemistry 33:3342–3349
Matsuzaki K, Murase O, Fujii N, Miyajima K (1996) An antimicrobial peptide, magainin 2, induced rapid flip-flop of phospholipids coupled with pore formation and peptide translocation. Biochemistry 35:11361–11368
Matsuzaki K, Sugishita K, Fujii N, Miyajima K (1995) Molecular basis for membrane selectivity of an antimicrobial peptide, magainin 2. Biochemistry 34:3423–3429
Mizuhara T, Saha K, Moyano DF, Kim CS, Yan B, Kim YK, Rotello VM (2015) Acylsulfonamide-functionalized zwitterionic gold nanoparticles for enhanced cellular uptake at tumor pH. Angew Chem Int Ed Engl 54:6567–6570
Nakatsuji T, Gallo RL (2012) Antimicrobial peptides: old molecules with new ideas. J Invest Dermatol 132:887–895
Nguyen VP, Alves DS, Scott HL, Davis FL, Barrera FN (2015) A novel soluble peptide with pH-responsive membrane insertion. Biochemistry 54:6567–6575
Onyango JO, Chung MS, Eng CH, Klees LM, Langenbacher R, Yao L, An M (2015) Noncanonical amino acids to improve the pH response of pHLIP insertion at tumor acidity. Angew Chem Int Ed Engl 54:3658–3663
Papo N, Shai Y (2003) New lytic peptides based on the d, l-amphipathic helix motif preferentially kill tumor cells compared to normal cells. Biochemistry 42:9346–9354
Papo N, Shai Y (2005) Host defense peptides as new weapons in cancer treatment. Cell Mol Life Sci 62:784–790
Riedl S, Zweytick D, Lohner K (2011) Membrane-active host defense peptides-challenges and perspectives for the development of novel anticancer drugs. Chem Phys Lipids 164:766–781
Schweizer F (2009) Cationic amphiphilic peptides with cancer-selective toxicity. Eur J Pharmacol 625:190–194
Song J, Zhang W, Kai M, Chen J, Liang R, Zheng X, Li G, Zhang B, Wang K, Zhang Y, Yang Z, Ni J, Wang R (2013) Design of an acid-activated antimicrobial peptide for tumor therapy. Mol Pharm 10:2934–2941
Sun CY, Shen S, Xu CF, Li HJ, Liu Y, Cao ZT, Yang XZ, Xia JX, Wang J (2015) Tumor acidity-sensitive polymeric vector for active targeted siRNA delivery. J Am Chem Soc 137:15217–15224
Tachi T, Epand RF, Epand RM, Matsuzaki K (2002) Position-dependent hydrophobicity of the antimicrobial magainin peptide affects the mode of peptide–lipid interactions and selective toxicity. Biochemistry 41:10723–10731
Takeshima K, Chikushi A, Lee K-K, Yonehara S, Matsuzaki K (2003) Translocation of analogues of the antimicrobial peptides magainin and buforin across human cell membranes. J Biol Chem 278:1310–1315
Uematsu N, Matsuzaki K (2000) Polar Angle as a Determinant of amphipathic α-helix-lipid interactions: a model peptide study. Biophys J 79:2075–2083
Vallabhapurapu SD, Blanco VM, Sulaiman MK, Vallabhapurapu SL, Chu Z, Franco RS, Qi X (2015) Variation in human cancer cell external phosphatidylserine is regulated by flippase activity and intracellular calcium. Oncotarget 6:34375–34388
Wenk MR, Seelig J (1998) Magainin 2 amide interaction with lipid membranes: calorimetric detection of peptide binding and pore formation. Biochemistry 37:3909–3916
Winiski AP, McLaughlin AC, McDaniel RV, Eisenberg M, McLaughlin S (1986) An experimental test of the discreteness-of-charge effect in positive and negative lipid bilayers. Biochemistry 25:8206–8214
Zhao Z, Meng H, Wang N, Donovan MJ, Fu T, You M, Chen Z, Zhang X, Tan W (2013) A controlled-release nanocarrier with extracellular pH value driven tumor targeting and translocation for drug delivery. Angew Chem Int Ed Engl 52:7487–7491
Zhou NE, Kay CM, Hodges RS (1992) Synthetic model proteins. Positional effects of interchain hydrophobic interactions on stability of two-stranded α-helical coiled-coils. J Biol Chem 267:2664–2670
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Wakabayashi, N., Yano, Y., Kawano, K. et al. A pH-dependent charge reversal peptide for cancer targeting. Eur Biophys J 46, 121–127 (2017). https://doi.org/10.1007/s00249-016-1145-y
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DOI: https://doi.org/10.1007/s00249-016-1145-y