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PCL-1, a Trypsin-Resistant Peptide, Exerts Potent Activity Against Drug-Resistant Bacteria

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Abstract

Antimicrobial peptides (AMPs), which hold tremendous promise in overcoming the emergence of drug resistance, are limited in wide clinical applications due to their instability, especially against trypsin. Herein, we designed six peptide mutants based on the cathelicidin CATHPb2, followed by screening. Pb2-1, which showed the best activity against drug-resistant bacteria among these mutants, was selected to be combined with the trypsin inhibitory loop ORB-C to obtain two hybrid peptides: PCL-1 and Pb2-1TI. Notably, both of the hybrid peptides exhibited a remarkable enhancement in trypsin resistance compared with Pb2-1. The tests showed that PCL-1 displayed broad-spectrum antimicrobial activity that was superior to that of Pb2-1TI. In addition, PCL-1 had relatively lower cytotoxicity than Pb2-1TI towards the L02 and HaCaT cell lines and negligible hemolysis, as well as tolerance to high concentrations of salt, extreme pH, and temperature variations. In vivo, PCL-1 effectively improved the survival rate of mice that were systemically infected with drug-resistant Escherichia coli through efficient bacterial clearance from the blood and organs. With regard to mode of action, PCL-1 damaged the integrity of the bacterial cell membrane and attached to the membrane surface while bound to bacterial genomic DNA to eventually kill the bacteria. Altogether, the trypsin-resistant peptide PCL-1 is expected to be a candidate for the clinical treatment of bacterial infections.

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Funding

This work was funded by the National Key Research and Development Program of China (2018YFA0902000), the National Natural Science Foundation of China (No. 81803591), the National Science and Technology Major Project Foundation of China (2019ZX09721001-004–005), the fellowship of China Postdoctoral Science Foundation (2020T130723), the Natural Science Foundation of Jiangsu Province of China (No. BK20180563; No. BK20201327), and the Basic Scientific Research Business Expense Project of China Pharmaceutical University (No. 2632021ZD07).

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  1. Haomin Wu and Pengfei Xu contributed equally to this work.

Authors and Affiliations

  1. School of Life Science and Technology, China Pharmaceutical University, 639 Longmian Avenue, Nanjing, 211198, Jiangsu, China

    Haomin Wu, Pengfei Xu, Ya Huang, Liping Wang, Xinyue Ye, Xiaowei Huang, Lingman Ma & ChangLin Zhou

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  1. Haomin Wu
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Correspondence to Lingman Ma or ChangLin Zhou.

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All experimental procedures were approved by the animal care and management committee of China Pharmaceutical University (2020–03-001).

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Wu, H., Xu, P., Huang, Y. et al. PCL-1, a Trypsin-Resistant Peptide, Exerts Potent Activity Against Drug-Resistant Bacteria. Probiotics & Antimicro. Prot. 13, 1467–1480 (2021). https://doi.org/10.1007/s12602-021-09801-8

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