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Transformation of the naturally occurring frog skin peptide, alyteserin-2a into a potent, non-toxic anti-cancer agent

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Abstract

Alyteserin-2a (ILGKLLSTAAGLLSNL.NH2) is a cationic, amphipathic α-helical cell-penetrating peptide, first isolated from skin secretions of the midwife toad Alytes obstetricans. Structure–activity relationships were investigated by synthesizing analogs of alyteserin-2a in which amino acids on the hydrophobic face of the helix were replaced by l-tryptophan and amino acids on the hydrophilic face were replaced by one or more l-lysine or d-lysine residues. The Trp-containing peptides display increased cytotoxic activity against non-small cell lung adenocarcinoma A549 cells (up to 11-fold), but hemolytic activity against human erythrocytes increases in parallel. The potency of the N15K analog against A549 cells (LC50 = 13 μM) increases sixfold relative to alyteserin-2a and the therapeutic index (ratio of LC50 for erythrocytes and tumor cells) increases twofold. Incorporation of a d-Lys11 residue into the N15K analog generates a peptide that retains potency against A549 cells (LC50 = 15 μM) but whose therapeutic index is 13-fold elevated relative to the native peptide. [G11k, N15K] alyteserin-2a is also active against human hepatocarcinoma HepG2 cells (LC50 = 26 μM), breast adenocarcinoma MDA-MB-231 cells (LC50 = 20 μM), and colorectal adenocarcinoma HT-29 cells (LC50 = 28 μM). [G11k, N15K] alyteserin-2a, in concentrations as low as 1 μg/mL, significantly (P < 0.05) inhibits the release of the immune-suppressive cytokines IL-10 and TGF-β from unstimulated and concanavalin A-stimulated peripheral blood mononuclear cells. The data suggest a strategy of increasing the cationicity while reducing the helicity of naturally occurring amphipathic α-helical peptides to generate analogs with improved cytotoxicity against tumor cells but decreased activity against non-neoplastic cells.

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Acknowledgments

This work was supported by a University Grant (G00000900) and a Faculty Support Grant (NP-10-12/103) from the United Arab Emirates University and by a grant from the Terry Fox for Cancer Research.

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Medicine and Health Sciences, United Arab Emirates University, 17666, Al Ain, United Arab Emirates

    J. Michael Conlon, Milena Mechkarska & Manju Prajeep

  2. Department of Pharmacology, Faculty of Medicine and Health Sciences, United Arab Emirates University, 17666, Al Ain, United Arab Emirates

    Kholoud Arafat & Samir Attoub

  3. Department of Biochemistry, Faculty of Medical Sciences, University of Kragujevac, Kragujevac, Serbia

    Milan Zaric

  4. Center for Molecular Medicine, Faculty of Medicine, University of Kragujevac, Kragujevac, Serbia

    Miodrag L. Lukic

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  1. J. Michael Conlon
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Correspondence to J. Michael Conlon.

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Conlon, J.M., Mechkarska, M., Prajeep, M. et al. Transformation of the naturally occurring frog skin peptide, alyteserin-2a into a potent, non-toxic anti-cancer agent. Amino Acids 44, 715–723 (2013). https://doi.org/10.1007/s00726-012-1395-7

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  • DOI: https://doi.org/10.1007/s00726-012-1395-7

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