In Vitro & In Vivo Studies on Identifying and Designing Temporin-1CEh from the Skin Secretion of Rana chensinensis as the Optimised Antibacterial Prototype Drug.

Zhuming Ye,Xiaowei Zhou,Xiaoling Chen,Xinping Xi,Lei Wang,Chengbang Ma,Mei Zhou,Tianbao Chen,Yu Zai,Hang Fai Kwok

doi:10.3390/pharmaceutics14030604

Abstract

Amphibian skin secretion is an ideal source of antimicrobial peptides that are difficult to induce drug resistance to due to their membrane-targeting mechanism as a new treatment scheme. In this study, a natural antimicrobial peptide Temporin-1CEh was identified by molecular cloning and mass spectrometry from the skin secretions of the Chinese forest frog (Rana chensinensis). Through the study of the structure and biological activity, it was found that Temporin-1CEh was a helical peptide from the Temporin family, and possessed good anti-Gram-positive bacteria activity through the mechanism of membrane destruction. Seven analogues were further designed to obtain broad-spectrum antimicrobial activity and higher stability in different physiological conditions. The results showed that T1CEh-KKPWW showed potent antibacterial activity with significantly increasing the activity against Gram-negative bacteria in vitro and in vivo with low haemolysis. In addition, T1CEh-KKPWW2 showed high sensitivity to the pH, serum or salts conditions, which applied a branched structure to allow the active units of the peptide to accumulate. Even though the haemolytic activity was increased, the stable antibacterial activity made this novel analogue meet the conditions to become a potential candidate in future antimicrobial and antibiofilm applications.

Highlights

IntroductionThe occurrence of multiple drug resistance of pathogens is increasing, and even some pathogens can evade the treatment of all clinically used antibiotics [1]
Introduction published maps and institutional affilNowadays, the occurrence of multiple drug resistance of pathogens is increasing, and even some pathogens can evade the treatment of all clinically used antibiotics [1]
Through the bioinformatic alignment using the NCBI-BLAST programme, the precursor peptide shows a high degree of similarity to preproTemporin-1CEa and other six Temporin family prepropeptides from Rana frogs (Figure 2)

Summary

Introduction

The occurrence of multiple drug resistance of pathogens is increasing, and even some pathogens can evade the treatment of all clinically used antibiotics [1]. The severe situation of multidrug drug resistance makes it urgent to develop new antibiotics [2,3]. After decades of exploration and research on the natural peptide, more and more attention has been paid to their antimicrobial potential. They have a broad spectrum of antimicrobial activity, including gram-positive and gram-negative bacteria, fungi and viruses [4]. The bacteria and fungi develop resistance to antibiotics through the reduction of drug permeability to their biomembranes, an efflux of antibiotics molecules from the cell, modification of antimicrobial targets, or enzymatic hydrolysis and degradation [7–12]

Methods

Results

Conclusion