Design and Synthesis of Fluorescent Acyclic Nucleoside Phosphonates as Potent Inhibitors of Bacterial Adenylate Cyclases

0467409 - ÚOCHB 2017 RIV DE eng J - Journal Article
Břehová, Petra - Šmídková, Markéta - Skácel, Jan - Dračínský, Martin - Mertlíková-Kaiserová, Helena - Velasquez, M. P. S. - Watts, V. J. - Janeba, Zlatko
Design and Synthesis of Fluorescent Acyclic Nucleoside Phosphonates as Potent Inhibitors of Bacterial Adenylate Cyclases.
ChemMedChem. Roč. 11, č. 22 (2016), s. 2534-2546. ISSN 1860-7179. E-ISSN 1860-7187
R&D Projects: GA MV VG20102015046; GA MŠMT LO1302
Institutional support: RVO:61388963
Keywords : adenylate cyclase toxin * acyclic nucleoside phosphonates * anthranilic acid
Subject RIV: CC - Organic Chemistry
Impact factor: 3.225, year: 2016

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 the 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 sub-micromolar potency (IC50 values: 11-622nm). 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 in direct fluorescence with Cl-ANT-ANPpp having an ester linker was strictly calmodulin (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=12m.
Permanent Link: http://hdl.handle.net/11104/0265511