Methicillin-resistant Staphylococcus aureus (MRSA) poses a major threat to human health and food safety, especially when bacteria form biofilms or invade host cells, which may cause recurring infections. A new solution is therefore urgently needed. The antimicrobial peptide innate defense regulator (IDR)-1018 and its derived peptide 1018M showed promising antimicrobial and antibiofilm activities. Nevertheless, their antibacterial efficacy against intracellular MRSA and protease tolerance remains to be promoted. Therefore, we synthesized D-amino acid substitution peptides D1018 and D1018M. The antimicrobial activity against MRSA of these novel peptides was increased by 1-fold (D1018) or remained constant (D1018M) compared with L-amino acids peptides. Their bactericidal mechanisms involve cell wall destruction, membrane penetration, and genomic DNA disruption. As expected, the stability of D1018 and D1018M was increased by 2–32 times against pepsin, trypsin, and cathepsin K. In addition, by D-amino acids substitution, the antibiofilm ability of D1018 was increased by 1.6 times, and the anti-intracellular bacterial activity of D1018M was improved 3.2–5.7 orders of magnitude. These data indicated that D1018M is a potential antimicrobial candidate for recurring MRSA infections.
Ben HurD, KapachG, WaniNA, et al.Antimicrobial peptides against multidrug-resistant Pseudomonas aeruginosa biofilm from cystic fibrosis patients. J Med Chem, 2022; 65(13):9050–9062; doi: 10.1021/acs.jmedchem.2c00270
2.
BöttgerR, HoffmannR, KnappeD. Differential stability of therapeutic peptides with different proteolytic cleavage sites in blood, plasma and serum. PLoS One, 2017; 12(6):e0178943; doi: 10.1371/journal.pone.0178943
3.
BucciniDF, CardosoMH, FrancoOL. Antimicrobial peptides and cell-penetrating peptides for treating intracellular bacterial infections. Front Cell Infect Microbiol, 2020; 10:612931; doi: 10.3389/fcimb.2020.612931
4.
CasciaroB, LinQ, AfoninS, et al.Inhibition of Pseudomonas aeruginosa biofilm formation and expression of virulence genes by selective epimerization in the Peptide Esculentin-1a(1-21)NH2. FEBS J, 2019; 286(19):3874–3891; doi: 10.1111/febs.14940
5.
de BreijA, RioolM, CordfunkeRA, et al.The antimicrobial peptide SAAP-148 combats drug-resistant bacteria and biofilms. Sci Transl Med, 2018; 10(423):eaan4044; doi: 10.1126/scitranslmed.aan4044
6.
de BreijA, RioolM, KwakmanPHS, et al.Prevention of Staphylococcus aureus biomaterial-associated infections using a polymer-lipid coating containing the antimicrobial peptide OP-145. J Control Release, 2016; 222:1–8; doi: 10.1016/j.jconrel.2015.12.003
7.
de la Fuente-NúñezC, ReffuveilleF, HaneyEF, et al.Broad-spectrum anti-biofilm peptide that targets a cellular stress response. PLoS Pathog, 2014; 10(5):e1004152; doi: 10.1371/journal.ppat.1004152
8.
FjellCD, HissJA, HancockRE, et al.Designing antimicrobial peptides: Form follows function. Nat Rev Drug Discov, 2011; 11(1):37–51; doi: 10.1038/nrd3591
9.
GomarascaM, MartinsTFC, GreuneL, et al.Bacterium-derived cell-penetrating peptides deliver gentamicin to kill intracellular pathogens. Antimicrob Agents Chemother, 2017; 61(4):e02545-16; doi: 10.1128/aac.02545-16
HøibyN, BjarnsholtT, GivskovM, et al.Antibiotic resistance of bacterial biofilms. Int J Antimicrob Agents, 2010; 35(4):322–332; doi: 10.1016/j.ijantimicag.2009.12.011
12.
HongX, QinJ, LiT, et al.Staphylococcal Protein A promotes colonization and immune evasion of the epidemic healthcare-associated MRSA ST239. Front Microbiol, 2016; 7:951; doi: 10.3389/fmicb.2016.00951
13.
JialeZ, JianJ, XinyiT, et al.Design of a novel antimicrobial peptide 1018M Targeted ppGpp to inhibit MRSA biofilm formation. AMB Express, 2021; 11(1):49; doi: 10.1186/s13568-021-01208-6
14.
KamaruzzamanNF, FirdessaR, GoodL. Bactericidal effects of Polyhexamethylene Biguanide against intracellular Staphylococcus aureus EMRSA-15 and USA 300. J Antimicrob Chemother, 2016; 71(5):1252–1259; doi: 10.1093/jac/dkv474
15.
KapilS, SharmaV. D-Amino acids in antimicrobial peptides: A potential approach to treat and combat antimicrobial resistance. Can J Microbiol, 2021; 67(2):119–137; doi: 10.1139/cjm-2020-0142
16.
KharaJS, PriestmanM, UhíaI, et al.Unnatural Amino Acid analogues of membrane-active helical peptides with anti-mycobacterial activity and improved stability. J Antimicrob Chemother, 2016; 71(8):2181–2191; doi: 10.1093/jac/dkw107
17.
KościuczukEM, LisowskiP, JarczakJ, et al.Cathelicidins: Family of antimicrobial peptides. A review. Mol Biol Rep, 2012; 39(12):10957–10970; doi: 10.1007/s11033-012-1997-x
18.
KumarP, KizhakkedathuJN, StrausSK. Antimicrobial peptides: Diversity, mechanism of action and strategies to improve the activity and biocompatibility in vivo. Biomolecules, 2018; 8(1):4; doi: 10.3390/biom8010004
19.
LevineDP. Vancomycin: A history. Clin Infect Dis, 2006; 42 (Suppl 1):S5–S12; doi: 10.1086/491709
20.
LiH, AnuwongcharoenN, MalikAA, et al.Roles of D-Amino acids on the bioactivity of host defense peptides. Int J Mol Sci, 2016; 17(7):1023; doi: 10.3390/ijms17071023
21.
LiW, SeparovicF, O’Brien-SimpsonNM, et al.Chemically modified and conjugated antimicrobial peptides against superbugs. Chem Soc Rev, 2021; 50(8):4932–4973; doi: 10.1039/d0cs01026j
22.
LinB, HungA, SingletonW, et al.The effect of tailing lipidation on the bioactivity of antimicrobial peptides and their aggregation tendency. Aggregate, 2023; 4(4):e329; doi: 10.1002/agt2.329
23.
LlarrullLI, FisherJF, MobasheryS. Molecular basis and phenotype of methicillin resistance in Staphylococcus aureus and insights into new beta-lactams that meet the challenge. Antimicrob Agents Chemother, 2009; 53(10):4051–4063; doi: 10.1128/AAC.00084-09
24.
LundinP, JohanssonH, GuterstamP, et al.Distinct uptake routes of cell-penetrating peptide conjugates. Bioconjug Chem, 2008; 19(12):2535–2542; doi: 10.1021/bc800212j
25.
MonackDM, MuellerA, FalkowS. Persistent bacterial infections: The interface of the pathogen and the host immune system. Nat Rev Microbiol, 2004; 2(9):747–765; doi: 10.1038/nrmicro955
26.
NelsonR. Antibiotic development pipeline runs dry. New drugs to fight resistant organisms are not being developed, experts say. Lancet, 2003; 362(9397):1726–1727; doi: 10.1016/s0140-6736(03)14885-4
27.
OngZY, WiradharmaN, YangYY. Strategies employed in the design and optimization of synthetic antimicrobial peptide amphiphiles with enhanced therapeutic potentials. Adv Drug Deliv Rev, 2014; 78:28–45; doi: 10.1016/j.addr.2014.10.013
28.
QiuS, ZhuR, ZhaoY, et al.Antimicrobial activity and stability of protonectin with D-Amino acid substitutions. J Pept Sci, 2017; 23(5):392–402; doi: 10.1002/psc.2989
29.
SuzukiK, MurakamiT, HuZ, et al.Human host defense Cathelicidin Peptide LL-37 enhances the lipopolysaccharide uptake by Liver Sinusoidal Endothelial Cells without Cell Activation. J Immunol, 2016; 196(3):1338–1347; doi: 10.4049/jimmunol.1403203
30.
TanP, FuH, MaX. Design, optimization, and nanotechnology of Antimicrobial Peptides: From exploration to applications. Nano Today, 2021; 39:101229; doi: 10.1016/j.nantod.2021.101229
31.
UhligT, KyprianouT, MartinelliFG, et al.The emergence of peptides in the pharmaceutical business: From exploration to exploitation. EuPA Open Proteomics, 2014; 4:58–69; doi: 10.1016/j.euprot.2014.05.003
WangD, HaapasaloM, GaoY, et al.Antibiofilm peptides against biofilms on titanium and hydroxyapatite surfaces. Bioact Mater, 2018a;3(4):418–425; doi: 10.1016/j.bioactmat.2018.06.002
34.
WangX, TengD, MaoR, et al.Combined systems approaches reveal a multistage mode of action of a marine antimicrobial peptide against pathogenic Escherichia coli and its protective effect against bacterial peritonitis and endotoxemia. Antimicrob Agents Chemother, 2016; 61(1):e01056-16; doi: 10.1128/aac.01056-16
35.
WangX, WangX, TengD, et al.Increased intracellular activity of MP1102 and NZ2114 against Staphylococcus aureus in vitro and in vivo. Sci Rep, 2018b;8(1):4204; doi: 10.1038/s41598-018-22245-5
36.
WątłyJ, MillerA, KozłowskiH, et al.Peptidomimetics – an infinite reservoir of metal binding motifs in metabolically stable and biologically active molecules. J Inorg Biochem, 2021; 217:111386; doi: 10.1016/j.jinorgbio.2021.111386
37.
WieczorekM, JenssenH, KindrachukJ, et al.Structural studies of a peptide with immune modulating and direct antimicrobial activity. Chem Biol, 2010; 17(9):970–980; doi: 10.1016/j.chembiol.2010.07.007
38.
WiegandI, HilpertK, HancockREW. Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances. Nat Protoc, 2008; 3(2):163–175; doi: 10.1038/nprot.2007.521
39.
WuR, DongX, WangQ, et al.D1018 with higher stability and excellent lipopolysaccharide binding affinity has potent anti-bacterial and anti-inflammatory activity. Front Microbiol, 2022; 13:1010017; doi: 10.3389/fmicb.2022.1010017
40.
YangN, LiuX, TengD, et al.Antibacterial and detoxifying Activity of NZ17074 Analogues with Multi-Layers of Selective Antimicrobial Actions against Escherichia coli and Salmonella enteritidis. Sci Rep, 2017; 7(1):3392; doi: 10.1038/s41598-017-03664-2
YeZ, ZhuX, AcostaS, et al.Self-assembly dynamics and antimicrobial activity of all L- and D-Amino acid enantiomers of a designer peptide. Nanoscale, 2018; 11(1):266–275; doi: 10.1039/C8NR07334A
43.
YinLM, LeeS, MakJSW, et al.Differential binding of L- vs. D-isomers of cationic antimicrobial peptides to the biofilm exopolysaccharide alginate. Protein Pept Lett, 2013; 20(8):843–847; doi: 10.2174/0929866511320080001
44.
ZhongC, GouS, LiuT, et al.Study on the effects of different dimerization positions on biological activity of partial D-Amino acid substitution analogues of Anoplin. Microb Pathog, 2020; 139:103871; doi: 10.1016/j.micpath.2019.103871
