Design and Synthesis of Fluorescent Acyclic Nucleoside Phosphonates as Potent Inhibitors of Bacterial Adenylate Cyclases.
Design and Synthesis of Fluorescent Acyclic Nucleoside Phosphonates as Potent Inhibitors of Bacterial Adenylate Cyclases.
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DOI：
10.1002/cmdc.201600439
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发表时间：
2016-11-21
期刊：
影响因子：
3.4
通讯作者：
Janeba, Zlatko
中科院分区：
文献类型：
--
作者：
Brehova, Petra;Smidkova, Marketa;Skacel, Jan;Dracinsky, Martin;Mertlikova-Kaiserova, Helena;Velasquez, Monica P. Soto;Watts, Val J.;Janeba, Zlatko
百日咳杆菌腺苷酸环化酶毒素（ACT）和炭疽杆菌水肿因子（EF）是具有腺苷酸环化酶（AC）活性的关键毒力因子，分别在百日咳和炭疽的发病机制中起重要作用。目前迫切需要开发具有潜在抗菌治疗前景的细菌AC的有效和选择性抑制剂，并研究其与靶酶的分子相互作用。设计并合成了新型荧光5-氯邻氨基苯甲酰取代的无环核苷膦酸酯（Cl-ANT-ANP），其二磷酸酯（Cl-ANT-ANPpp）作为竞争性ACT和EF抑制剂，具有亚微摩尔效力（IC 50值11 - 622 nM）。荧光实验表明，Cl-ANT-ANPpp类似物结合ACT活性位点和对接研究表明，Cl-ANT基团与Phe 306和Leu 60相互作用。有趣的是，具有酯接头的Cl-ANT-ANPpp的直接荧光的增加是严格依赖于CaM的，而具有酰胺接头的Cl-ANT-ANPpp类似物在与ACT结合后，即使在不存在CaM的情况下也增加荧光。这种结合对结构修饰的依赖性可以在未来设计细菌AC的有效抑制剂中利用。此外，一种双酰胺酯前药形式的Cl-ANT-ANP能够抑制B。巨噬细胞中的百日咳ACT活性，IC 50 = 12 µM。
Bordetella pertussis adenylate cyclase toxin (ACT) and Bacillus anthracis edema factor (EF) are key virulence factors with adenylate cyclase (AC) activity that substantially contribute to the pathogenesis of whooping cough and anthrax, respectively. There is an urgent need to develop potent and selective inhibitors of bacterial ACs with prospects for development of potential antibacterial therapeutics and to study their molecular interactions with the target enzymes. Novel fluorescent 5-chloroanthraniloyl-substituted acyclic nucleoside phosphonates (Cl-ANT-ANPs) were designed and synthesized in the form of their diphosphates (Cl-ANT-ANPpp) as competitive ACT and EF inhibitors with submicromolar potency (IC50 values 11 – 622 nM). Fluorescence experiments indicated that Cl-ANT-ANPpp analogues bind to the ACT active site and docking studies suggested that the Cl-ANT group interacts with Phe306 and Leu60. Interestingly, the increase of direct fluorescence with Cl-ANT-ANPpp having an ester linker was strictly CaM-dependent, whereas Cl-ANT-ANPpp analogues with an amide linker, upon binding to ACT, increased the fluorescence even in the absence of CaM. Such a dependence of binding on structural modification could be exploited in the future design of potent inhibitors of bacterial ACs. Furthermore, one Cl-ANT-ANP in the form of a bisamidate prodrug was able to inhibit B. pertussis ACT activity in macrophage cells with IC50 = 12 µM.
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影响因子：
4.8
作者：
Gille, A;Lushington, GH;Seifert, R
通讯作者：
Seifert, R
DOI：
10.1124/jpet.113.207449
发表时间：
2013-11-01
影响因子：
3.5
作者：
Conley, Jason M.;Brand, Cameron S.;Watts, Val J.
通讯作者：
Watts, Val J.
影响因子：
4.9
作者：
Birkus, Gabriel;Wang, Ruth;McDermott, Martin
通讯作者：
McDermott, Martin
影响因子：
3.1
作者：
Hradilova, Ludmila;Polakova, Monika;Petrus, Ladislav
通讯作者：
Petrus, Ladislav
影响因子：
11.4
作者：
Guo, Q;Shen, YQ;Tang, WJ
通讯作者：
Tang, WJ