A Membrane Curvature Modulated Lipopeptide to Broadly Combat Multidrug-Resistant Bacterial Pneumonia with Low Resistance Risk

Abstract

The extensive spread of multidrug resistance to Gram-negative bacteria has become a huge threat to human health, where peptide-based antibacterial agents have emerged as a powerful star weapon. Here we report a lipopeptide (LP-20) constructed nanomicelle with a different antibacterial mechanism of membrane curvature modulation, which induced dynamic membrane fission resulting in acceleration and enhancement of antibacterial activity to clinically isolated ESKAPE strains, including multidrug-resistant (MDR) pathogens. The minimum inhibitory concentration was reduced to 2−10 μM, and the minimum duration for killing was shortened to less than an hour by LP-20. This is an improvement over antimicrobial peptides and traditional antibiotics, such as ciprofloxacin and tetracycline, significantly enhancing antibacterial activity for MDR, and we observed no acquisition of resistance for one month. This accelerated germicidal mechanism was attributed to multitargeting with lipopolysaccharides, phosphoethanolamine, phosphatidylglycerol, and cardiolipin, and the synergetic interactions induced a high curvature of the bacterial membrane, which facilitated simultaneously efficient damage to both inner and outer membrane. The LP-20 effectively prolonged the lifetime of myositis mice with Escherichia coli MDR and pneumonia mice with Klebsiella pneumoniae through a hepatic metabolism with ignorable toxicity. This study provides critical information for the fabrication of lipopeptide-based nano-antibiotics for the efficient control of intractable MDR caused by Gram-negative pathogens.

Membrane-Active Peptides Attack Cell Membranes in a Lipid-Regulated Curvature-Generating Mode

“…22 These experimental findings, combined with our simulations and energybased theoretical analyses, aid in comprehending the mechanism behind recently observed MAP-induced morphological changes in live cells. 17,21,22 Our work clearly demonstrates that the simplistic membrane models utilized in previous studies are inadequate for simulating MAP activity, as certain behaviors such as changes in membrane curvature may be easily overlooked. These findings have significant implications for enhancing the biological efficacy of MAPs while minimizing their cytotoxicity through adjustments to their membrane-dependent attack mode.…”

“…76 The recent experiments conducted by Zou et al have demonstrated that peptide-induced membrane fission can also contribute to membrane damage. 22 These findings suggest that the leakage of contents and cellular death in bacteria under the influence of MAPs may not be solely attributed to pore formation. Classical models of pore formation primarily focus on the poration effect of peptides, while complex interactions between peptides and lipids are often overlooked.…”

“…21 Recently, experiments have shown that lipopeptide assembly (LP-20) can induce high curvature and dynamic fission of bacterial membranes, ultimately enhancing antibacterial activity. 22 Membrane curvature change is a crucial characteristic of many cellular processes (Figure S1), 23−29 lying MAPs' membrane activity and, particularly, explore their potential cell membrane-specific effects. Molecular dynamics (MD) simulations are a potent tool for providing atomic-level information on peptide (or protein)− membrane interactions.…”

Peptide Supramolecular Hydrogels with Sustained Release Ability for Combating Multidrug-Resistant Bacteria

“…Extended-spectrum β-lactamases (ESBL) and methicillin-resistant Staphylococcus aureus (MRSA) are multidrug-resistant bacterial pathogens with strong infectivity and high lethality. 23 The minimal inhibitory concentration (MIC) of Pep 6 against MRSA DN65 and ESBL T1T was 25 and 20.88 ± 7.21 μg/mL, respectively. In contrast, TPA showed no notable antimicrobial activity, with MIC values above 400 μg/mL (Table S1).…”

“…The bioactivity of Hydrogel-RL was measured against the increasing emergence and dissemination of resistant bacterial pathogens. Extended-spectrum β-lactamases (ESBL) and methicillin-resistant Staphylococcus aureus (MRSA) are multidrug-resistant bacterial pathogens with strong infectivity and high lethality . The minimal inhibitory concentration (MIC) of Pep 6 against MRSA DN65 and ESBL T1T was 25 and 20.88 ± 7.21 μg/mL, respectively.…”