Structure-Activity Approach in the Reactivation of Tabun-Phosphorylated Human Acetylcholinesterase with Bispyridinium para-Aldoximes

Kovarik, Zrinka; Čalić, Maja; Šinko, Goran; Bosak, Anita

doi:10.2478/v10004-007-0013-7

Abstract

We investigated interactions of bispyridinium para-aldoximes N,N'-(propano)bis(4-hydroxyiminomethyl) pyridinium bromide (TMB-4), N,N'-(ethano)bis(4-hydroxyiminomethyl)pyridinium methanosulphonate (DMB-4), and N,N'-(methano)bis(4-hydroxyiminomethyl)pyridinium chloride (MMB-4) with human erythrocyte acetylcholinesterase phosphorylated by tabun. We analysed aldoxime conformations to determine the flexibility of aldoxime as an important feature for binding to the acetylcholinesterase active site. Tabun-inhibited human erythrocyte acetylcholinesterase was completely reactivated only by the most flexible bispyridinium aldoxime - TMB-4 with a propylene chain between two rings. Shorter linkers than propylene (methylene or ethylene) as in MMB-4 and DMB-4 did not allow appropriate orientation in the active site, and MMB-4 and DMB-4 were not efficient reactivators of tabun-phosphorylated acetylcholinesterase. Since aldoximes are also reversible inhibitors of native acetylcholinesterase, we determined dissociation constants and their protective index against acetylcholinesterase inactivation by tabun.

References

Gray AP. Design and structure-activity relationships of antidotes to organophosphorus anticholinesterase agents. Drug Metab Rev 1984;15, 557-89.10.3109/03602538409029973
Search in Google Scholar
Stojiljković MP, Jokanović M. Pyridinium oximes: rationale for their selection as causal antidotes against organophosphate poisonings and current solutions for auto-injectors. Arh Hig Rada Toksikol 2006;57:435-43.
Search in Google Scholar
Čalić M, Lucić Vrdoljak A, Radić B, Jelić D, Jun D, Kuča K, Kovarik Z. In vitro and in vivo evaluation of pyridinium aldoximes: mode of interaction with acetylcholinesterase, effect on tabun- and soman-poisoned mice and their cytotoxicity. Toxicology 2006;219:85-96.10.1016/j.tox.2005.11.003
Search in Google Scholar
Lucić Vrdoljak A, Čalić M, Radić B, Berend S, Jun D, Kuča K, Kovarik Z. Pretreatment with pyridinium aldoximes improves antidotal therapy against tabun poisoning. Toxicology 2006;228:41-50.10.1016/j.tox.2006.08.012
Search in Google Scholar
Simeon-Rudolf V, Reiner E, Škrinjarić-Špoljar M, Radić B, Lucić A, Primožić I, Tomić S. Quinuclidinium-imidazolium compounds: synthesis, mode of interaction with acetylcholinesterase and effect upon soman intoxicated mice. Arch Toxicol 1998;72:289-95.10.1007/s002040050504
Search in Google Scholar
Škrinjarić-Špoljar M, Burger N, Lovrić J. Inhibition of acetylcholinesterase by three new pyridinium compounds and their effect on phosphonylation of the enzyme. J Enzym Inhib 1999;14:331-41.10.3109/14756369909030327
Search in Google Scholar
Kovarik Z, Bosak A, Šinko G, Latas T. Exploring active sites of cholinesterases by inhibition with bambuterol and haloxon. Croat Chem Acta 2003;76:63-7.
Search in Google Scholar
Šinko G, Bosak A, Kovarik Z, Simeon-Rudolf V. Structure-inhibition relationships in the interaction of butyrylcholinesterase with bambuterol, haloxon and their leaving groups. Chem-Biol Interact 2005;157-158:421-3.10.1016/j.cbi.2005.10.087
Search in Google Scholar
Ellman GL, Courtney KD, Andres VJr, Featherstone RM. New and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 1961;7:88-95.10.1016/0006-2952(61)90145-9
Search in Google Scholar
Eyer P, Worek F, Kiderlen D, Šinko G, Štuglin A, Simeon-Rudolf V, Reiner E. Molar absorption coefficients for the reduced Ellman reagent: reassessment. Anal Biochem 2003;312:224-7.10.1016/S0003-2697(02)00506-7
Search in Google Scholar
Šinko G, Čalić M, Kovarik Z. para- and ortho-Pyridinium aldoximes in reaction with acetylthiocholine. FEBS Lett 2006;580:3167-72.10.1016/j.febslet.2006.04.07016684539
Search in Google Scholar
Odžak R, Čalić M, Hrenar T, Primožić I, Kovarik Z. Evaluation of monoquaternary pyridinium aldoximes potency to reactivate tabun-inhibited human acetylcholinesterase. Toxicology 2006;233:85-96.10.1016/j.tox.2006.08.00316962227
Search in Google Scholar
Kovarik Z, Radić Z, Berman HA, Simeon-Rudolf V, Reiner E, Taylor P. Mutant cholinesterases possessing enhanced capacity for reactivation of their phosphonylated conjugates. Biochemistry 2004;43:3222-9.10.1021/bi036191a
Search in Google Scholar
Heilbronn E. In vitro reactivation and "aging" of tabun-inhibited blood cholinesterases studied with N-methyl-pyridinium-2-aldoxime methane sulphonate and N,N'-trimethylene bis (pyridinium-4-aldoxime) dibromide. Biochem Pharmacol 1963;12:25-36.10.1016/0006-2952(63)90006-6
Search in Google Scholar
Ashani Y, Bhattacharjee AK, Leader H, Saxena A, Doctor BP. Inhibition of cholinesterase with cationic phosphonyl oximes highlights distinctive properties of the charged pyridine groups of quaternary oxime reactivators. Biochem Pharmacol 2003;33:191-202.10.1016/S0006-2952(03)00204-1
Search in Google Scholar
Francisković L, Škrinjarić-Špoljar M, Reiner E. Interaction of imidazolium and pyridinium dioximes with human erythrocyte acetylcholinesterase. Chem-Biol Interact 1993;87:323-8.10.1016/0009-2797(93)90060-C
Search in Google Scholar
Kuča K, Patočka J, Cabal J. Reactivation of organophosphate inhibited acetylcholinesterase activity by α, ω-bis-(hydroxyiminomethylpyridinium)alkanes in vitro.J Appl Biomed 2003;1:207-11.10.32725/jab.2003.037
Search in Google Scholar
Worek F, Thiermann H, Szinicz L, Eyer P. Kinetic analysis of interactions between human acetylcholineserase, structurally different organophosphorus compounds and aldoximes. Biochem Pharmacol 2004;68:2237-48.10.1016/j.bcp.2004.07.038
Search in Google Scholar
Pang Y-P, Kollmeyer TM, Hong F, Lee J, Hammond PI, Haugabouk SP, Brimijoin S. Rational design of alkylene-linked bis-pyridiniumaldoximes as improved acetylcholinesterase reactivators. Chem Biol 2003;10:491-502.10.1016/S1074-5521(03)00126-1
Search in Google Scholar
Ekström F, Pang Y-P, Boman M, Artursson E, Akfur C, Börjegren S. Crystal structures of acetylcholinesterase in complex with HI-6, Ortho-7 and obidoxime: Structural basis for differences in the ability to reactivate tabun conjugates. Biochem Pharmacol 2006;72:597-607.10.1016/j.bcp.2006.05.02716876764
Search in Google Scholar
Bosak A, Primožić I, Oršulić M, Tomić S, Simeon-Rudolf V. Enantiomers of quinuclidin-3-ol derivates: resolution and interactions with human cholinesterases. Croat Chem Acta 2005;78:121-8.
Search in Google Scholar
Simeon V, Radić Z, Reiner E. Inhibition of cholinesterase by the aldoximes P2AM and toxogonin. Croat Chem Acta 1981;54:473-80.
Search in Google Scholar
Škrinjarić-Špoljar M, Simeon V, Reiner E, Krauthacker B. Bispyridinium compounds: inhibition of human erythrocyte acetylcholinesterase and protection of the enzyme against phosphylation. Acta Pharm Jugosl 1988;38:101-9.
Search in Google Scholar
Kovarik Z, Ciban N, Radić Z, Simeon-Rudolf V, Taylor P. Active site mutant acetylcholinesterase interactions with 2-PAM, HI-6, and DDVP. Biochem Biophys Res Commun 2006;342:973-8.10.1016/j.bbrc.2006.02.05616598855
Search in Google Scholar
Reiner E. Inhibition of acetylcholinesterase by 4,4'-bipyridine and its effect upon phosphylation of the enzyme. Croat Chem Acta 1986;59:925-31.
Search in Google Scholar

Structure-Activity Approach in the Reactivation of Tabun-Phosphorylated Human Acetylcholinesterase with Bispyridinium para-Aldoximes

Abstract

Paradigm

My account