Duda Addison M, Ma Helena R, Villalobos César A, Kuhn Sophia A, He Katherine, Seay Sarah R, Jackson Abigail C, Suh Christine M, Puccio Elena A, Anderson Deverick J, Fowler Vance G, You Lingchong, Franz Katherine J
Department of Chemistry, Duke University, Durham, NC 27710, USA.
Department of Biomedical Engineering, Duke University, Durham, NC 27710, USA.
bioRxiv. 2024 Aug 3:2024.08.02.606422. doi: 10.1101/2024.08.02.606422.
The rise of β-lactam resistance necessitates new strategies to combat bacterial infections. We purposefully engineered the β-lactam prodrug AcephPT to exploit β-lactamase activity to selectively suppress resistant bacteria producing extended-spectrum-β-lactamases (ESBLs). Selective targeting of resistant bacteria requires avoiding interaction with penicillin-binding proteins, the conventional targets of β-lactam antibiotics, while maintaining recognition by ESBLs to activate AcephPT only in resistant cells. Computational approaches provide a rationale for structural modifications to the prodrug to achieve this biased activity. We show AcephPT selectively suppresses gram-negative ESBL-producing bacteria in clonal populations and in mixed microbial cultures, with effective selectivity for both lab strains and clinical isolates expressing ESBLs. Time-course NMR experiments confirm hydrolytic activation of AcephPT exclusively by ESBL-producing bacteria. In mixed microbial cultures, AcephPT suppresses proliferation of ESBL-producing strains while sustaining growth of β-lactamase-non-producing bacteria, highlighting its potential to combat β-lactam resistance while promoting antimicrobial stewardship.
β-内酰胺耐药性的上升使得对抗细菌感染需要新的策略。我们特意设计了β-内酰胺前药AcephPT，以利用β-内酰胺酶活性来选择性地抑制产生超广谱β-内酰胺酶（ESBLs）的耐药细菌。选择性靶向耐药细菌需要避免与β-内酰胺抗生素的传统靶点青霉素结合蛋白相互作用，同时保持被ESBLs识别，以便仅在耐药细胞中激活AcephPT。计算方法为前药的结构修饰提供了理论依据，以实现这种偏向性活性。我们表明，AcephPT在克隆群体和混合微生物培养物中选择性地抑制产生革兰氏阴性ESBL的细菌，对表达ESBL的实验室菌株和临床分离株均具有有效的选择性。时间进程核磁共振实验证实，AcephPT仅由产生ESBL的细菌进行水解激活。在混合微生物培养物中，AcephPT抑制产生ESBL菌株的增殖，同时维持不产生β-内酰胺酶细菌的生长，突出了其在对抗β-内酰胺耐药性同时促进抗菌药物管理的潜力。
