title: Amidate prodrugs of 9-[2-(phosphonomethoxy)ethyl]adenine as inhibitors of adenylate cyclase toxin from Bordetella pertussis.
abstract: Adenylate cyclase toxin (ACT) is the key virulence factor of Bordetella pertussis that facilitates its invasion into the mammalian body. 9-[2-(Phosphonomethoxy)ethyl]adenine diphosphate (PMEApp), the active metabolite of the antiviral drug bis(POM)PMEA (adefovir dipivoxil), has been shown to inhibit ACT. The objective of this study was to evaluate six novel amidate prodrugs of PMEA, both phenyloxy phosphonamidates and phosphonodiamidates, for their ability to inhibit ACT activity in the J774A.1 macrophage cell line. The two phenyloxy phosphonamidate prodrugs exhibited greater inhibitory activity (50% inhibitory concentration [IC50] = 22 and 46 nM) than the phosphonodiamidates (IC50 = 84 to 3,960 nM). The inhibitory activity of the prodrugs correlated with their lipophilicity and the degree of their hydrolysis into free PMEA in J774A.1 cells. Although the prodrugs did not inhibit ACT as effectively as bis(POM)PMEA (IC50 = 6 nM), they were significantly less cytotoxic. Moreover, they all reduced apoptotic effects of ACT and prevented an ACT-induced elevation of intracellular [Ca(2+)]i. The amidate prodrugs were less susceptible to degradation in Caco-2 cells compared to bis(POM)PMEA, while they exerted good transepithelial permeability in this assay. As a consequence, a large amount of intact amidate prodrug is expected to be available to target macrophages in vivo. This feature makes nontoxic amidate prodrugs attractive candidates for further investigation as novel antimicrobial agents.
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
authors.forename: Markéta
authors.initials: M
authors.lastname: Šmídková
authors.forename: Alexandra
authors.initials: A
authors.lastname: Dvoráková
authors.forename: Eva
authors.initials: E
authors.lastname: Tloust'ová
authors.forename: Michal
authors.initials: M
authors.lastname: Česnek
authors.forename: Zlatko
authors.initials: Z
authors.lastname: Janeba
authors.forename: Helena
authors.initials: H
authors.lastname: Mertlíková-Kaiserová
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D037361:Adenylate Cyclase Toxin / Q000037:antagonists & inhibitors* / Q000378:metabolism; D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism / Q000494:pharmacology*; D001886:Bordetella pertussis / Q000187:drug effects* / Q000254:growth & development / Q000472:pathogenicity; D018938:Caco-2 Cells; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D006801:Humans; D020128:Inhibitory Concentration 50; D008264:Macrophages / Q000187:drug effects / Q000382:microbiology; D051379:Mice; D008826:Microbial Sensitivity Tests; D063065:Organophosphonates / Q000494:pharmacology; D011355:Prodrugs / Q000378:metabolism / Q000494:pharmacology*
chemical_list: C120290:9-(2-(phosphonomethoxy)ethyl)adenine diphosphate; D037361:Adenylate Cyclase Toxin; D000900:Anti-Bacterial Agents; D063065:Organophosphonates; D011355:Prodrugs; D000225:Adenine; C106812:adefovir dipivoxil