Flow-Based Fmoc-SPPS Preparation and SAR Study of Cathelicidin-PY Reveals Selective Antimicrobial Activity.
Shama Dissanayake, Junming He, Sung H Yang, Margaret A Brimble, Paul W R Harris, Alan J Cameron
Author Information
Shama Dissanayake: School of Chemical Sciences and School of Biological Sciences, The University of Auckland, 23 Symonds St., Auckland 1142, New Zealand.
Junming He: School of Chemical Sciences and School of Biological Sciences, The University of Auckland, 23 Symonds St., Auckland 1142, New Zealand.
Sung H Yang: School of Chemical Sciences and School of Biological Sciences, The University of Auckland, 23 Symonds St., Auckland 1142, New Zealand.
Margaret A Brimble: School of Chemical Sciences and School of Biological Sciences, The University of Auckland, 23 Symonds St., Auckland 1142, New Zealand. ORCID
Paul W R Harris: School of Chemical Sciences and School of Biological Sciences, The University of Auckland, 23 Symonds St., Auckland 1142, New Zealand.
Alan J Cameron: School of Chemical Sciences and School of Biological Sciences, The University of Auckland, 23 Symonds St., Auckland 1142, New Zealand. ORCID
Antimicrobial peptides (AMPs) hold promise as novel therapeutics in the fight against multi-drug-resistant pathogens. Cathelicidin-PY (NH-RKCNFLCKLKEKLRTVITSHIDKVLRPQG-COOH) is a 29-residue disulfide-cyclised antimicrobial peptide secreted as an innate host defence mechanism by the frog (PY) and reported to possess broad-spectrum antibacterial and antifungal properties, exhibiting low cytotoxic and low hemolytic activity. Herein, we detail the total synthesis of cathelicidin-PY using an entirely on-resin synthesis, including assembly of the linear sequence by rapid flow Fmoc-SPPS and iodine-mediated disulfide bridge formation. By optimising a synthetic strategy to prepare cathelicidin-PY, this strategy was subsequently adapted to prepare a bicyclic head-to-tail cyclised derivative of cathelicidin-PY. The structure-activity relationship (SAR) of cathelicidin-PY with respect to the -terminally positioned disulfide was further probed by preparing an alanine-substituted linear analogue and a series of lactam-bridged peptidomimetics implementing side chain to side chain cyclisation. The analogues were investigated for antimicrobial activity, secondary structure by circular dichroism (CD), and stability in human serum. Surprisingly, the disulfide bridge emerged as non-essential to antimicrobial activity and secondary structure but was amenable to synthetic modification. Furthermore, the synthetic AMP and multiple analogues demonstrated selective activity towards Gram-negative pathogen in physiologically relevant concentrations of divalent cations.
