Ayerbe-Algaba Rafael, Bayó Nuria, Verdú Ester, Parra-Millán Raquel, Seco Jesús, Teixidó Meritxell, Pachón Jerónimo, Giralt Ernest, Smani Younes
Clinical Unit of Infectious Diseases, Microbiology and Preventive Medicine, University Hospital Virgen del Rocío, Seville, Spain.
Institute of Biomedicine of Seville (IBiS), University Hospital Virgen del Rocío/Spanish National Research Council (CSIC)/University of Seville, Seville, Spain.
Front Microbiol. 2021 Feb 12;12:634323. doi: 10.3389/fmicb.2021.634323. eCollection 2021.
Previously, we identified that a cyclic hexapeptide AOA-2 inhibited the interaction of Gram-negative bacilli (GNB) like , , and to host cells thereby preventing the development of infection and in a murine sepsis peritoneal model. In this work, we aimed to evaluate a library of AOA-2 derivatives in order to improve the effect of AOA-2 against GNB infections. Ten AOA-2 derivatives were synthetized for the assays. Their toxicities to human lung epithelial cells (A549 cells) for 24 h were evaluated by determining the A549 cells viability using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The effect of these peptide derivatives and AOA-2 at 250, 125, 62.5, and 31.25 μg/mL on the attachment of ATCC 17978, PAO1 and ATCC 25922 strains to A549 cells was characterized by adherence and viability assays. None of the 10 derivatives showed toxicity to A549 cells. RW01 and RW06 have reduced more the adherence of ATCC 17978, PAO1 and ATCC 2599 strains to A549 cells when compared with the original compound AOA-2. Moreover, both peptides have increased slightly the viability of infected A549 cells by PAO1 and ATCC 25922 than those observed with AOA-2. Finally, RW01 and RW06 have potentiated the activity of colistin against ATCC 17978 strain in the same level with AOA-2. The optimization program of AOA-2 has generated two derivatives (RW01 and RW06) with best effect against interaction of GNB with host cells, specifically against and .
此前，我们发现一种环状六肽AOA - 2可抑制诸如大肠杆菌、铜绿假单胞菌和肺炎克雷伯菌等革兰氏阴性杆菌（GNB）与宿主细胞的相互作用，从而在小鼠脓毒症腹膜模型中预防感染的发展。在这项研究中，我们旨在评估AOA - 2衍生物文库，以提高AOA - 2对GNB感染的疗效。合成了10种AOA - 2衍生物用于实验。通过使用3 -（4,5 - 二甲基噻唑 - 2 - 基）- 2,5 - 二苯基四氮唑溴盐法测定A549细胞活力，评估它们对人肺上皮细胞（A549细胞）24小时的毒性。通过黏附及活力测定，表征了这些肽衍生物和AOA - 2在250、125、62.5和31.25μg/mL浓度下对大肠杆菌ATCC 17978、铜绿假单胞菌PAO1和肺炎克雷伯菌ATCC 25922菌株黏附到A549细胞上的影响。这10种衍生物均未显示对A549细胞有毒性。与原始化合物AOA - 2相比，RW01和RW06更多地降低了大肠杆菌ATCC 17978、铜绿假单胞菌PAO1和肺炎克雷伯菌ATCC 2599菌株对A549细胞的黏附。此外，与AOA - 2相比，这两种肽均使被铜绿假单胞菌PAO1和肺炎克雷伯菌ATCC 25922感染的A549细胞的活力略有增加。最后，RW01和RW06增强了黏菌素对大肠杆菌ATCC 17978菌株的活性，与AOA - 2处于同一水平。AOA - 2的优化程序产生了两种对GNB与宿主细胞相互作用，特别是对大肠杆菌和铜绿假单胞菌具有最佳效果的衍生物（RW01和RW06）。
