Abstract
Aim:
To investigate whether the conjugation of magainin II (MG2), an antimicrobial peptides (AMPs), to the tumor-homing peptide bombesin could enhance its cytotoxicity in tumor cells.
Methods:
A magainin II-bombesin conjugate (MG2B) was constructed by attaching magainin II (MG2) to bombesin at its N-terminus. The peptides were synthesized using Fmoc-chemistry. The in vitro cytotoxicity of the peptide in cancer cells was quantitatively determined using the CCK-8 cell counting kit. Moreover, the in vivo antitumor effect of the peptide was determined in tumor xenograft models.
Results:
The IC50 of MG2B for cancer cells (10–15 μmol/L) was at least 10 times lower than the IC50 of unconjugated MG2 (125 μmol/L). Moreover, the binding affinity of MG2B for cancer cells was higher than that of unconjugated MG2. In contrast, conjugation to a bombesin analog lacking the receptor-binding domain failed to increase the cytotoxicity of MG2, suggesting that bombesin conjugation enhances the cytotoxicity of MG2 in cancer cells through improved binding. Indeed, MG2B selectively induced cell death in cancer cells in vitro with the IC50 ranging from 10 to 15 μmol/L, which was about 6–10 times lower than the IC50 for normal cells. MG2B (20 mg/kg per day, intratumorally injected for 5 d) also exhibited antitumor effects in mice bearing MCF-7 tumor grafts. The mean weights of tumor grafts in MG2B- and PBS-treated mice were 0.21±0.05 g and 0.59±0.12 g, respectively.
Conclusion:
The results suggest that conjugation of AMPs to bombesin might be an alternative approach for targeted cancer therapy.
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References
Mader JS, Hoskin DW . Cationic antimicrobial peptides as novel cytotoxic agents for cancer treatment. Expert Opin Investig Drugs 2006; 15: 933–46.
Lehmann J, Retz M, Sidhu SS, Suttmann H, Sell M, Paulsen F, et al. Antitumor activity of the antimicrobial peptide magainin II against bladder cancer cell lines. Eur Urol 2006; 50: 141–7.
Bhutia SK, Maiti TK . Targeting tumors with peptides from natural sources. Trends Biotechnol 2008; 26: 210–7.
Hoskin DW, Ramamoorthy A . Studies on anticancer activities of antimicrobial peptides. Biochim Biophys Acta 2008; 1778: 357–75.
Papo N, Seger D, Makovitzki A, Kalchenko V, Eshhar Z, Degani H, et al. Inhibition of tumor growth and elimination of multiple metastases in human prostate and breast xenografts by systemic inoculation of a host defense-like lytic peptide. Cancer Res 2006; 66: 5371–8.
Conlon JM, Kolodziejek J, Nowotny N . Antimicrobial peptides from the skins of North American frogs. Biochim Biophys Acta 2009; 1788: 1556–63.
Fernandez DI, Gehman JD, Separovic F . Membrane interactions of antimicrobial peptides from Australian frogs. Biochim Biophys Acta 2009; 1788: 1630–8.
Zasloff M . Magainins, a class of antimicrobial peptides from Xenopus skin: isolation, characterization of two active forms, and partial cDNA sequence of a precursor. Proc Natl Acad Sci USA 1987; 84: 5449–53.
Cruciani RA, Barker JL, Zasloff M, Chen HC, Colamonici O . Antibiotic magainins exert cytolytic activity against transformed cell lines through channel formation. Proc Natl Acad Sci USA 1991; 88: 3792–6.
Jacob L, Zasloff M . Potential therapeutic applications of magainins and other antimicrobial agents of animal origin. Ciba Found Symp 1994; 186: 197–216; discussion 216–23.
Baker MA, Maloy WL, Zasloff M, Jacob LS . Anticancer efficacy of Magainin2 and analogue peptides. Cancer Res 1993; 53: 3052–7.
Matsuzaki K, Nakamura A, Murase O, Sugishita K, Fujii N, Miyajima K . Modulation of magainin 2-lipid bilayer interactions by peptide charge. Biochemistry 1997; 36: 2104–11.
Liu Y, Steiniger SC, Kim Y, Kaufmann GF, Felding-Habermann B, Janda KD . Mechanistic studies of a peptidic GRP78 ligand for cancer cell-specific drug delivery. Mol Pharm 2007; 4: 435–47.
Nishimura S, Takahashi S, Kamikatahira H, Kuroki Y, Jaalouk DE, O'Brien S, et al. Combinatorial targeting of the macropinocytotic pathway in leukemia and lymphoma cells. J Biol Chem 2008; 283: 11752–62.
Laakkonen P, Zhang L, Ruoslahti E . Peptide targeting of tumor lymph vessels. Ann N Y Acad Sci 2008; 1131: 37–43.
Corti A, Curnis F, Arap W, Pasqualini R . The neovasculature homing motif NGR: more than meets the eye. Blood 2008; 112: 2628–35.
Zwanziger D, Beck-Sickinger AG . Radiometal targeted tumor diagnosis and therapy with peptide hormones. Curr Pharm Des 2008; 14: 2385–400.
Khan IU, Beck-Sickinger AG . Targeted tumor diagnosis and therapy with peptide hormones as radiopharmaceuticals. Anticancer Agents Med Chem 2008; 8: 186–99.
Okarvi SM . Peptide-based radiopharmaceuticals and cytotoxic conjugates: potential tools against cancer. Cancer Treat Rev 2008; 34: 13–26.
Anastasi A, Erspamer V, Bucci M . Isolation and structure of bombesin and alytesin, two analogous active peptides from the skin of the European amphibians Bombesina and Alytes. Experientia 1971; 27: 166–9.
Reubi JC, Wenger S, Schmuckli-Maurer J, Schaer JC, Gugger M . Bombesin receptor subtypes in human cancers: detection with the universal radioligand (125)I-[D-TYR(6), beta-ALA(11), PHE(13), NLE(14)] bombesin(6–14). Clin Cancer Res 2002; 8: 1139–46.
Cornelio DB, Roesler R, Schwartsmann G . Gastrin-releasing peptide receptor as a molecular target in experimental anticancer therapy. Ann Oncol 2007; 18: 1457–66.
de Visser M, Verwijnen SM, de Jong M . Update: improvement strategies for peptide receptor scintigraphy and radionuclide therapy. Cancer Biother Radiopharm 2008; 23: 137–57.
Engel JB, Keller G, Schally AV, Halmos G, Hammann B, Nagy A . Effective inhibition of experimental human ovarian cancers with a targeted cytotoxic bombesin analogue AN-215. Clin Cancer Res 2005; 11: 2408–15.
Smith CJ, Volkert WA, Hoffman TJ . Radiolabeled peptide conjugates for targeting of the bombesin receptor superfamily subtypes. Nucl Med Biol 2005; 32: 733–40.
Laakkonen P, Akerman ME, Biliran H, Yang M, Ferrer F, Karpanen T, et al. Antitumor activity of a homing peptide that targets tumor lymphatics and tumor cells. Proc Natl Acad Sci USA 2004; 101: 9381–6.
Rege K, Patel SJ, Megeed Z, Yarmush ML . Amphipathic peptide-based fusion peptides and immunoconjugates for the targeted ablation of prostate cancer cells. Cancer Res 2007; 67: 6368–75.
Skerlavaj B, Gennaro R, Bagella L, Merluzzi L, Risso A, Zanetti M . Biological characterization of two novel cathelicidin-derived peptides and identification of structural requirements for their antimicrobial and cell lytic activities. J Biol Chem 1996; 271: 28375–81.
Risso A, Braidot E, Sordano MC, Vianello A, Macrì F, Skerlavaj B, et al. BMAP-28, an antibiotic peptide of innate immunity, induces cell death through opening of the mitochondrial permeability transition pore. Mol Cell Biol 2002; 22: 1926–35.
Imura Y, Choda N, Matsuzaki K . Magainin 2 in action: distinct modes of membrane permeabilization in living bacterial and mammalian cells. Biophys J 2008; 95: 5757–65.
Westerhoff HV, Hendler RW, Zasloff M, Juretić D . Interactions between a new class of eukaryotic antimicrobial agents and isolated rat liver mitochondria. Biochim Biophys Acta 1989; 975: 361–9.
Wang G, Li X, Wang Z . APD2: the updated antimicrobial peptide database and its application in peptide design. Nucleic Acids Res 2009; 37: D933–7.
Suttmann H, Retz M, Paulsen F, Harder J, Zwergel U, Kamradt J, et al. Antimicrobial peptides of the Cecropin-family show potent antitumor activity against bladder cancer cells. BMC Urol 2008; 8: 5.
Pastan I, Hassan R, FitzGerald DJ, Kreitman RJ . Immunotoxin treatment of cancer. Annu Rev Med 2007; 58: 221–37.
Acknowledgements
This work was supported by the National Basic Research Program of China (2009CB522401), and the National Natural Science Foundation of China (81072566).
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Liu, S., Yang, H., Wan, L. et al. Enhancement of cytotoxicity of antimicrobial peptide magainin II in tumor cells by bombesin-targeted delivery. Acta Pharmacol Sin 32, 79–88 (2011). https://doi.org/10.1038/aps.2010.162
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DOI: https://doi.org/10.1038/aps.2010.162
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