Wang Yingyu, Liao Mingrui, Ge Tianhao, Ding Ke, Hughes Arwel, Li Peixun, Lu Jian R
Biological Physics Laboratory, Department of Physics and Astronomy, School of Natural Science, The University of Manchester, Oxford Road, Manchester M13 9PL, U.K.
ISIS Pulsed Neutron & Muon Source, STFC Rutherford Appleton Laboratory, Harwell Campus, Didcot OX11 0QX, U.K.
ACS Appl Mater Interfaces. 2025 Aug 27;17(34):47894-47905. doi: 10.1021/acsami.5c07879. Epub 2025 Aug 6.
Antimicrobial peptides (AMP) offer an attractive alternative to antibiotics in the global fight against antibiotic resistance. AMPs can impose fast structural disruptions to microbial membranes and kill pathogens by causing the leakage of their internal contents, making it less likely for pathogens to develop resistance. However, current AMPs still suffer from various drawbacks including weak efficacy, unacceptable toxicity, and side effects. This work seeks to design a group of amphiphilic AMPs based on the α2 sequence of a Dengue viral capsid protein to address the challenge of ineffective membrane disruptions of AMPs. The design was also inspired by well-studied G(IIKK)I-NH (G) for broad-spectrum antimicrobial actions. All designed Dengue viral-inspired peptides displayed lower minimum inhibition concentrations and faster time-dependent killing than G, with the fastest DVP-3 (RIFRAIRRIARFIR) achieving complete killing within 10 min. Fluorescence assays of AMP binding to bacterial membranes revealed varying degrees of membrane permeability change, depolarization, and leakage. Model inner membrane (IM) and outer membrane (OM) of Gram-negative bacteria facilitated leakage assay and neutron reflection, linking membrane binding and disruptions with antimicrobial behaviors. The findings reveal that DVP-3 could cause more effective disruptions to the bacterial OM than to the IM, consistent with its potent antimicrobial efficacy and rapid dynamic killing. This work offers new insights into how the newly designed AMPs destabilize bacterial membranes to improve antimicrobial performance and the combined approach allows effective AMP evaluation to overcome bacterial resistance.
抗菌肽（AMP）在全球对抗抗生素耐药性的斗争中为抗生素提供了一种有吸引力的替代方案。抗菌肽可对微生物膜造成快速的结构破坏，并通过导致其内部内容物泄漏来杀死病原体，从而降低病原体产生耐药性的可能性。然而，目前的抗菌肽仍存在各种缺点，包括疗效不佳、毒性不可接受以及副作用等。这项工作旨在基于登革热病毒衣壳蛋白的α2序列设计一组两亲性抗菌肽，以应对抗菌肽膜破坏效果不佳的挑战。该设计还受到了对具有广谱抗菌作用的G（IIKK）I-NH（G）深入研究的启发。所有设计的受登革热病毒启发的肽都比G表现出更低的最低抑菌浓度和更快的时间依赖性杀伤作用，其中最快的DVP-3（RIFRAIRRIARFIR）在10分钟内实现完全杀伤。抗菌肽与细菌膜结合的荧光测定揭示了不同程度的膜通透性变化、去极化和泄漏。革兰氏阴性菌的模型内膜（IM）和外膜（OM）促进了泄漏测定和中子反射，将膜结合和破坏与抗菌行为联系起来。研究结果表明，DVP-3对细菌外膜的破坏作用比对内膜更有效，这与其强大的抗菌功效和快速的动态杀伤作用一致。这项工作为新设计的抗菌肽如何破坏细菌膜以提高抗菌性能提供了新的见解，并且这种综合方法允许进行有效的抗菌肽评估以克服细菌耐药性。
