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Design and synthesis of oligo-lipidated arginyl peptide (OLAP) dimers with enhanced physicochemical activity, peptide stability and their antimicrobial actions against MRSA infections

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Abstract

Multi-drug resistant pathogens have been of increasing concern today. There is an urgent need for the discovery of more potent antibiotics. Cationic antimicrobial peptides (CAMPs) are known to be effective antimicrobial agents against resistant pathogens. However, poor activity under physiological conditions is one of the major limitations of CAMPS in clinical applications. In this study, a series of oligo-lipidated arginyl peptide OLAP dimers comprised of a saturated fatty acid chain (with m number of carbon units) and p repeating units of arginyl fatty acid chains (with n number of carbon units) were designed and studied for their antimicrobial activities as well as their physico-chemical property in various physiological conditions, such as in human serum albumin and high salt conditions. Our results showed that OLAP-11 exhibits potent antimicrobial activity against Gram-positive bacteria with improved physico-chemical activity in various physiological conditions. OLAP-11 is also less susceptible to human serum and trypsin degradation. The HPLC–MS analysis showed that the lipid-arginine bond is very stable. SYTOX Green assay and scanning electron microscopy both show that the OLAP-11 killed bacteria via inner membrane disruption. In addition, OLAP-11 is inner membrane targeting, making it difficult for bacteria to develop resistance. Overall, the design of the OLAP dimers provides an alternative approach to improve the physicochemical activity, peptide stability of CAMPs with potent inner membrane disruption and low in vitro toxicity to increase their potential for clinical applications in the future.

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Abbreviations

OLAP:

Oligo-lipidated arginyl peptide

CAMP:

Cationic antimicrobial peptide

HSA:

Human serum albumin

MIC:

Minimum inhibitory concentration

MRSA:

Methicillin-resistant Staphylococcus aureus

DOPE:

1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine

DOPG:

1,2-Dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol)

CL:

Cardiolipin

OAK:

Oligo-acyl-lysine OAK

CAMHB:

Cationic-adjusted Mueller–Hinton broth

CFU:

Colony forming units

RBC:

Red blood cells

ANSA:

8-(Phenylamino)-1-naphtha-lenesulfonic acid

MDR:

Multi-drug resistant

AMP:

Antimicrobial peptide

HPLC–MS:

High-performance liquid chromatography-mass spectrometry

OAK:

Oligo-acyl-lysine

CLSI:

Clinical and Laboratory Standards Institute

TSA:

Trypticase soy agar

MHA:

Mueller–Hinton agar

HEPES:

(4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid)

LUV:

Large unilamellar vehicle

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Acknowledgements

This work was supported by the National Medical Research Council NMRC/CBRG/0080/2015 and NMRC/TCR/R1012. This work is also supported by the Singhealth Foundation SHF/FG691S/2016.

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Authors and Affiliations

  1. Singapore Eye Research Institute, The Academia, 20 College Road, Discovery Tower Level 6, Singapore, 169856, Singapore

    Jun-Jie Koh, Shuimu Lin, Wendy Wan Ling Sin, Zhi Hao Ng, Do Yun Jung, Roger W. Beuerman & Shouping Liu

  2. School of Materials Science and Engineering, South China University of Technology, Guangzhou, 510641, China

    Shuimu Lin

  3. SRP Neuroscience and Behavioural Disorders, Duke-NUS Medical School, Singapore, 169857, Singapore

    Roger W. Beuerman & Shouping Liu

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  1. Jun-Jie Koh
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  2. Shuimu Lin
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The manuscript was written with contributions from all the authors. All the authors approved the final version of the manuscript.

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Correspondence to Roger W. Beuerman or Shouping Liu.

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This article does not contain any studies with human participants performed by any of the authors.

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All procedures performed in studies involving animals were in accordance with the ethical standards of the SingHealth IACUC in accordance with the guidelines of the Association for Research in Vision and Ophthalmology (ARVO) at which the studies were conducted.

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Handling Editor: J. D. Wade.

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Koh, JJ., Lin, S., Sin, W.W.L. et al. Design and synthesis of oligo-lipidated arginyl peptide (OLAP) dimers with enhanced physicochemical activity, peptide stability and their antimicrobial actions against MRSA infections. Amino Acids 50, 1329–1345 (2018). https://doi.org/10.1007/s00726-018-2607-6

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