Caputo Alessia, Sartini Sara, Levati Elisabetta, Minato Ilaria, Elisi Gian Marco, Di Stasi Adriana, Guillou Catherine, Goekjian Peter G, Garcia Pierre, Gueyrard David, Bach Stéphane, Comte Arnaud, Ottonello Simone, Rivara Silvia, Montanini Barbara
Laboratory of Biochemistry and Molecular Biology, Department of Chemistry, Life Sciences and Environmental Sustainability, University of Parma, 43124 Parma, Italy.
Interdepartmental Research Centre Biopharmanet-Tec, University of Parma, 43124 Parma, Italy.
Antibiotics (Basel). 2022 Oct 21;11(10):1449. doi: 10.3390/antibiotics11101449.
Bacterial resistance represents a major health problem worldwide and there is an urgent need to develop first-in-class compounds directed against new therapeutic targets. We previously developed a drug-discovery platform to identify new antimicrobials able to disrupt the protein-protein interaction between the β' subunit and the σ initiation factor of bacterial RNA polymerase, which is essential for transcription. As a follow-up to such work, we have improved the discovery strategy to make it less time-consuming and more cost-effective. This involves three sequential assays, easily scalable to a high-throughput format, and a subsequent in-depth characterization only limited to hits that passed the three tests. This optimized workflow, applied to the screening of 5360 small molecules from three synthetic and natural compound libraries, led to the identification of six compounds interfering with the β'-σ interaction, and thus was capable of inhibiting promoter-specific RNA transcription and bacterial growth. Upon supplementation with a permeability adjuvant, the two most potent transcription-inhibiting compounds displayed a strong antibacterial activity against with minimum inhibitory concentration (MIC) values among the lowest (0.87-1.56 μM) thus far reported for β'-σ PPI inhibitors. The newly identified hit compounds share structural feature similarities with those of a pharmacophore model previously developed from known inhibitors.
细菌耐药性是全球范围内的一个主要健康问题，迫切需要开发针对新治疗靶点的一流化合物。我们之前开发了一个药物发现平台，以识别能够破坏细菌RNA聚合酶β'亚基与σ起始因子之间蛋白质-蛋白质相互作用的新型抗菌药物，这种相互作用对于转录至关重要。作为此项工作的后续，我们改进了发现策略，使其耗时更短、成本效益更高。这包括三个连续的检测方法，可轻松扩展至高通量形式，随后仅对通过这三项测试的命中物进行深入表征。这种优化的工作流程应用于从三个合成和天然化合物库中筛选5360个小分子，从而鉴定出六种干扰β'-σ相互作用的化合物，因此能够抑制启动子特异性RNA转录和细菌生长。在添加通透性佐剂后，两种最有效的转录抑制化合物对[具体细菌名称未给出]显示出强大的抗菌活性，其最低抑菌浓度（MIC）值处于迄今为止报道的β'-σ蛋白-蛋白相互作用抑制剂中最低水平（0.87-1.56μM）。新鉴定出的命中化合物与先前从已知抑制剂开发的药效团模型具有结构特征相似性。
