Discovery of Inhibitors of 4′-Phosphopantetheine Adenylyltransferase (PPAT) To Validate PPAT as a Target for Antibacterial Therapy
Discovery of Inhibitors of 4′-Phosphopantetheine Adenylyltransferase (PPAT) To Validate PPAT as a Target for Antibacterial Therapy
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DOI：
10.1128/aac.01661-13
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发表时间：
2013-12-01
影响因子：
4.9
通讯作者：
Thresher, Jason
中科院分区：
文献类型：
--
作者：
de Jonge, Boudewijn L. M.;Walkup, Grant K.;Thresher, Jason
通过对阿斯利康化合物文库的高通量筛选，鉴定了4‘-磷酸丙氨酸腺苷转移酶(PPAT)的抑制剂。其中一个系列，环烷基嘧啶，对几个物种的PPAT同工酶有抑制作用，其中对革兰氏阳性物种的酶的抑制作用最强。对肺炎链球菌和金黄色葡萄球菌PPAT的抑制模式研究表明，这一系列的代表是与磷酸铁氨酸竞争而与ATP不竞争的可逆抑制剂，与酶-ATP复合体结合。采用基于结构的设计对该系列的效价进行了优化，实现了对革兰氏阳性菌细胞生长的抑制。作用模式研究，使用具有靶向测序的抗性突变体的产生以及过度表达PPAT的构建物，证明生长抑制是由于抑制PPAT所致。在小鼠肺部和大腿感染模型中证实了对细菌负荷的影响，但在考虑将这些化合物投入临床开发之前，需要进一步优化给药要求和化合物的性质。这些研究证实了PPAT是一种新的抗菌治疗靶点。
Inhibitors of 4'-phosphopantetheine adenylyltransferase (PPAT) were identified through high-throughput screening of the AstraZeneca compound library. One series, cycloalkyl pyrimidines, showed inhibition of PPAT isozymes from several species, with the most potent inhibition of enzymes from Gram-positive species. Mode-of-inhibition studies with Streptococcus pneumoniae and Staphylococcus aureus PPAT demonstrated representatives of this series to be reversible inhibitors competitive with phosphopantetheine and uncompetitive with ATP, binding to the enzyme-ATP complex. The potency of this series was optimized using structure-based design, and inhibition of cell growth of Gram-positive species was achieved. Mode-of-action studies, using generation of resistant mutants with targeted sequencing as well as constructs that overexpress PPAT, demonstrated that growth suppression was due to inhibition of PPAT. An effect on bacterial burden was demonstrated in mouse lung and thigh infection models, but further optimization of dosing requirements and compound properties is needed before these compounds can be considered for progress into clinical development. These studies validated PPAT as a novel target for antibacterial therapy.