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: 11-622 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 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 =12 μm.

Department of Medicinal Chemistry and Molecular Pharmacology College of Pharmacy Purdue University 575 Stadium Mall Drive West Lafayette IN 47907 USA

The Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences Flemingovo nám 2 166 10 Prague 6 Czech Republic

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Diavatopoulos DA, Cummings CA, Schouls LM, Brinig MM, Relman DA, Mooi FR. Plos Pathog. 2005;1:e45. PubMed PMC

Pertussis vaccines: WHO position paper: Weekly Epidemiological Record. 2010;85:385–400. PubMed

Kilgore PE, Salim AM, Zervos MJ, Schitt HJ. Clin. Microbiol. Rev. 2016;29:449–486. PubMed PMC

Tiwari T, Murphy TV, Moran J. [acccessed October 2015];Morbidity and Mortality Weekly Report. 2005 Dec 9; http://www.cdc.gov/mmwr/preview/mmwrhtml/rr5414a1.htm. PubMed

Wood N, McIntyre P. Pediatr. Resp. Rev. 2008;9:201–212. PubMed

Bartkus JM, Juni BA, Ehresmann K, Miller CA, Sanden GN, Cassiday PK, Saubolle M, Lee B, Long J, Harrison AR, Jr, Besser JM. J. Clin. Microbiol. 2003;41:1167–1172. PubMed PMC

Wendelboe AM, Rie AV, Salmaso S, Englund JA. Pediatr. Infect. Dis. J. 2005;24:S58–S61. PubMed

Ladant D, Ullmann A. Trends Microbiol. 1999;7:172–176. PubMed

Confer DL, Eaton JW. Science. 1982;217:948–950. PubMed

Glaser P, Sakamoto H, Bellalou J, Ullmann A, Danchin A. EMBO J. 1988;7:3997–4004. PubMed PMC

Mock M, Ullmann A. Trends Microbiol. 1993;1:187–192. PubMed

Ahuja N, Kumar P, Bhatnagar R. Crit. Rev. Microbiol. 2004;30:187–196. PubMed

Shen Y, Zhukovskaya NL, Zimmer MI, Soelaiman S, Bergson P, Wang CR, Gibbs CS, Tang WJ. Proc. Natl. Acad. Sci. USA. 2004;101:3242–3247. PubMed PMC

Martín C, Gómez-Bilbao G, Ostolaza H. J.Biol. Chem. 2010;285:357–364. PubMed PMC

Fisher R, Masin J, Bumba L, Pospisilova E, Fayolle C, Basler M, Sadilkova L, Adkins I, Kamanova J, Cerny J, Konopasek I, Osicka R, Laclerc C, Sebo P. PLoS Pathog. 2012;8(4):e1002580. PubMed PMC

De Clercq A, Holý E, Rosenberg I, Sakuma T, Balzarini J, Maudgal PC. Nature. 1986;323:464–467. PubMed

De Clercq E, Holý A. Nat. Rev. Drug Discovery. 2005;4:928–940. PubMed

De Clercq E. Med. Res. Rev. 2013;33:1278–1303. PubMed

Reiser H, Wang J, Chong L, Watkins WJ, Ray AS, Shibata R, Birkus G, Cihlar T, Wu S, Li B, Liu X, Henne IN, Wolfgang GHI, Desai M, Rhodes GR, Fridland A, Lee WA, Plunkett W, Vail D, Thamm DH, Jeraj R, Tumas DB. Clin. Cancer Res. 2008;14:2824–2832. PubMed

Zídek Z, Potměšil P, Holý A. Toxicol. Appl. Pharmacol. 2003;192:246–253. PubMed

Keough DT, Hocková D, Rejman D, Špaček P, Vrbková S, Krečmerová M, Eng WS, Jans H, West NP, Naesens LM, de Jersey J, Guddat LW. J. Med. Chem. 2013;56:6967–6984. PubMed

Eng WS, Hocková D, Špaček P, Janeba Z, West NP, Woods K, Naesens LMJ, Keough DT, Guddat LW. J. Med. Chem. 2015;58:4822–4838. PubMed

Keough DT, Hocková D, Holý A, Naesens LM, Skinner-Adams TS, de Jersey J, Guddat LW. J. Med. Chem. 2009;52:4391–4399. PubMed

Hocková D, Keough DT, Janeba Z, Wang TH, de Jersey J, Guddat LW. J. Med. Chem. 2012;55:6209–6223. PubMed

Keough DT, Špaček P, Hocková D, Tichý T, Vrbková S, Slavětínská L, Janeba Z, Naesens L, Edstein MD, Chavchich M, Wang TH, de Jersey J, Guddat LW. J. Med. Chem. 2013;56:2513–2526. PubMed

Keough DT, Hocková D, Janeba Z, Wang T, Naesens L, Edstein MD, Chavchich M, Guddat LW. J. Med. Chem. 2015;58:827–846. PubMed

Kaiser MM, Hocková D, Wang T-H, Draèínský M, Poštová-Slavìtínská L, Procházková E, Edstein MD, Chavchich M, Keough DT, Guddat LW, Janeba Z. ChemMedChem. 2015;10:1707–1723. PubMed

Zídek Z, Franková D, Holý A. Antimicrob. Agents Chemother. 2001;45:3381–3386. PubMed PMC

Zídek Z, Franková D, Holý A. Int. J. Immunopharmacol. 2000;22:1121–1129. PubMed

Potměšil P, Krečmerová M, Kmoníčková E, Holý A, Zídek Z. Eur. J. Pharmacol. 2006;540:191–199. PubMed

Guo Q, Shen Y, Lee Y-S, Gibbs CS, Mrksich M, Tang W. EMBO J. 2005;24:3190–3201. PubMed PMC

Shen YQ, Zhukovskaya NL, Zimmer MI, Soelaiman S, Bergson P, Wang CR, Gibbs CS, Tang WJ. Proc. Natl. Acad. Sci. USA. 2004;101:3242–3247. PubMed PMC

Shoshani I, Laux WHG, Périgaud Ch, Gosselin G, Johnson RA. J. Biol. Chem. 1999;274:34742–34744. PubMed

Česnek M, Jansa P, Šmídková M, Mertlíková-Kaiserová H, Dračínský M, Brust TF, Pávek P, Trejtnar F, Watts VJ, Janeba Z. ChemMedChem. 2015;10:1351–1364. PubMed

Šmídková M, Dvořáková A, Tloušťová E, Česnek M, Janeba Z, Mertlíková-Kaiserová H. Antimicrob. Agents Chemother. 2014;58:664–671. PubMed PMC

Pradere U, Garnier-Amblard EC, Coats SJ, Amblard F, Schinazi RF. Chem. Rev. 2014;114:9154–9218. PubMed PMC

Gille A, Seifert R. J. Biol. Chem. 2003;278:12672–12679. PubMed

Gille A, Lushington GH, Mou TC, Doughty MB, Johnson RA, Seifert R. J. Biol. Chem. 2004;279:19955–19969. PubMed

Seifert R, Dove S. Trends Microbiol. 2012;20:343–351. PubMed

Hiratsuka T. J. Biol. Chem. 1985;260:4784–4790. PubMed

Göttle M, Dove S, Steindel P, Shen Y, Tang W, Geduhn J, König B, Seifert R. Mol. Pharmacol. 2007;72:526–535. PubMed

Holý A. Collect. Czech. Chem. Commun. 1978;43:2054–2031.

Kuhn K, Owen DJ, Bader B, Wittinghofer A, Kuhlmann J, Waldmann H. J. Am. Chem. Soc. 2001;123:1023–1035. PubMed

Hradilová L, Poláková M, Dvořáková B, Hajdúch M, Petruš L. Carboh. Res. 2012;361:1–6. PubMed

Chen JJ, Cai X, Szostak JW. J. Am. Chem. Soc. 2009;131:2119–2121. PubMed PMC

Allison BD, Phuong VK, McAtee LC, et al. J. Med. Chem. 2006;49:6371–6390. PubMed

Twin H, Batey RA. Org. Lett. 2011;6:4913–4916. PubMed

Liu J, Deng X, Fitzgerald AE, Sales ZS, Vankatesan H, Mani S. Org. Biomol. Chem. 2011;9:2654–2660. PubMed

Krečmerová M, Masojídková M, Holý A. Collect. Czech. Chem. Commun. 2004;69:1889–1913.

Krečmerová M, Dračínský M, Hocková D, Holý A, Keough DT, Guddat LW. Bioorg. Med. Chem. 2012;20:1222–1230. PubMed

Vrbovská S, Holý A, Pohl R, Masojídková M. Collect. Czech. Chem. Commun. 2006;71:543–566.

Jansa P, Baszczynski O, Dračínský M, Votruba I, Zídek Z, Bahador G, Stepan G, Cihlar T, Mackman R, Holý A, Janeba Z. Eur. J. Med. Chem. 2011;46:3748–3754. PubMed

Holý A, Rosenberg I. Collect. Czech. Chem. Commun. 1987;52:2801–2809.

Moffatt JG, Khorana HG. J. Am. Chem. Soc. 1961;83:649–658.

Xu B, Stephens A, Kirschenheuter G, Greslin AF, Cheng X, Sennelo J, Cattaneo M, Zighetti ML, Chen A, Kim S, Kim HS, Bischofberger N, Cook G, Jacobson KA. J. Med. Chem. 2002;45:5694–5709. PubMed PMC

Kim HS, Barak D, Harden TK, Boyer JL, Jacobson KA. J. Med. Chem. 2001;44:3092–3108. PubMed PMC

Hackett M, Walker CB, Guo L, Gray MC, Van Cuyk S, Ullmann A, Shabonowith J, Hunt DF, Hewlett EL, Sebo P. J. Biol. Chem. 1995;270:20250–20253. PubMed

Iwaki M, Kamachi K, Konda T. Infect. Immun. 2000;68:3727–3730. PubMed PMC

Birkus G, Wang R, Liu X, Kutty N, MacArthur H, Cihlar T, Gibbs C, Swaminathan S, Lee W, McDemott M. Antimicrob. Agents Chemother. 2007;51:543–550. PubMed PMC

Luedtke CC, Andonian S, Igdoura S, Hermo L. J. Histochem. Cytochem. 2000;48:1131–1146. PubMed

Molecular Operating Environment (MOE) 2014:0901.

Chemical Computing Group Inc. 1010 Sherbooke St. West, Suite #910. Montreal, QC, Canada, H3A 2R7: 2016.

Šolínová V, Kaiser MM, Lukáč M, Janeba Z, Kašička V. J. Sep. Sci. 2014;37:295–303. PubMed

Conley JM, Brand CS, Bogard AS, Pratt EPS, Xu RQ, Hockerman GH, Ostrom RS, Dessauer CW, Watts VJ. J. Pharm. and Exp. Ther. 2013;347:276–287. PubMed PMC

Conley JM, Brust TF, Xu R, Burris KD, Watts VJ. J. Vis. Exp. 2014;83:e51218. PubMed PMC

Phosphonates and Phosphonate Prodrugs in Medicinal Chemistry: Past Successes and Future Prospects

Halogen-Dance-Based Synthesis of Phosphonomethoxyethyl (PME) Substituted 2-Aminothiazoles as Potent Inhibitors of Bacterial Adenylate Cyclases

Acyclic nucleoside phosphonates with 2-aminothiazole base as inhibitors of bacterial and mammalian adenylate cyclases