Zhu Yongjie, Li Bowen, Xu Wanying, Wang Yuanmengxue, Li Guoyu, Bi Chongpeng, Shan Anshan, Shao Changxuan
College of Animal Science and Technology, Northeast Agricultural University, Harbin 150030, China.
College of Animal Science and Technology, Northeast Agricultural University, Harbin 150030, China.
Drug Resist Updat. 2025 Mar;79:101183. doi: 10.1016/j.drup.2024.101183. Epub 2024 Dec 9.
The unstable antimicrobial activity of antimicrobial peptides (AMPs) under physiological conditions (especially the degradation instigated proteases) seems to be a persistent impediment for their successful implementation in clinical trials. Consequently, our objective was to devise AMP engineering frameworks that could sustain robust antibacterial efficacy within physiological environments.
In this work, we harvested AMPs with stable antimicrobial activity under the physiological barriers through the combination of idealized amphiphiles and trypsin inhibitors.
We screened and identified the lead peptides IK3-A and IK3-S, which showed potent activity against Gram-negative bacteria, including multidrug-resistant (MDR) bacteria, and exhibited promising biocompatibility with mammalian cells. Remarkably, IK3-A and IK3-S maintained sustained antibacterial potency under physiological salts, serum, and protease conditions. Furthermore, both IK3-A and IK3-S kill Gram-negative bacteria by attacking the bacterial cell membrane and inducing oxidative damage (at high concentrations). Crucially, IK3-A and IK3-S have optimal safety and efficacy in mice.
This is the first work to compare the effects of different trypsin inhibitors on the resistance of AMPs to protease hydrolysis on the same sequence platform. In conclusion, these findings provide guidance for the molecular design of AMPs with stable antibacterial activity under physiological conditions and facilitates the process of clinical translation of AMPs as antimicrobial biomaterials against MDR bacteria. Moreover, this may stimulate a more general interest in protease inhibitors as molecular scaffolds in the creation of highly stable peptide-based biomaterials.
抗菌肽（AMPs）在生理条件下不稳定的抗菌活性（尤其是蛋白酶引发的降解）似乎一直是其在临床试验中成功应用的障碍。因此，我们的目标是设计出能在生理环境中保持强大抗菌效力的AMPs工程框架。
在这项工作中，我们通过将理想化两亲物和胰蛋白酶抑制剂相结合，筛选出在生理屏障下具有稳定抗菌活性的AMPs。
我们筛选并鉴定出先导肽IK3 - A和IK3 - S，它们对革兰氏阴性菌（包括多重耐药菌）显示出强效活性，并且与哺乳动物细胞具有良好的生物相容性。值得注意的是，IK3 - A和IK3 - S在生理盐、血清和蛋白酶条件下保持持续的抗菌效力。此外，IK3 - A和IK3 - S均通过攻击细菌细胞膜并诱导氧化损伤（高浓度时）来杀死革兰氏阴性菌。至关重要的是，IK3 - A和IK3 - S在小鼠体内具有最佳的安全性和疗效。
这是第一项在同一序列平台上比较不同胰蛋白酶抑制剂对AMPs抗蛋白酶水解作用影响的研究。总之，这些发现为在生理条件下具有稳定抗菌活性的AMPs分子设计提供了指导，并促进了AMPs作为抗多重耐药菌抗菌生物材料的临床转化进程。此外，这可能会激发人们对蛋白酶抑制剂作为分子支架用于创建高度稳定的肽基生物材料产生更广泛的兴趣。
