The efficacy of 2-formyl benzoic acid in reactivating diazinon inhibited murine cholinesterase

Abstract

Oximes, as classical acetylcholinesterase (AChE) reactivators, have some pharmacokinetics/pharmacodynamics disadvantages. During the synthesis of non-oxime compounds, we encountered the compound 2-formylbenzoic acid (2-FBA) with promising in vitro and in vivo cholinesterase (ChE) reactivating properties in the acute exposure to diazinon (DZN). For in vitro experiments, the healthy mice serum and brain homogenate were freshly prepared and exposed to DZN (160 µg/mL). After 10 minutes, 2-FBA was added to the poisoned samples, and ChE activity was measured afterward. For the in vivo assay, the mice were poisoned with DZN subcutaneous (SC) injection (50 mg/kg), and after 1 hour, either 2-FBA or Pralidoxime (2-PAM) was injected intravenously (IV). After 3 h, ChE activity was measured in the serum and brain homogenate samples. The LD50 (IV) for 2-FBA in mice was measured as well. 2-FBA effectively reactivated the inhibited ChE in serum and brain homogenate samples in vitro. In the in vivo experiments, while 2-FBA could significantly reactivate the brain ChE even better than 2-PAM, they failed to reactivate the serum ChE by single IV injection. LD50 of 2-FBA was calculated to be 963 mg/kg. There were no general toxicity signs in any treatment groups. The in silico results support the potential ability of 2-FBA efficacy via possibly Witting reaction mechanism. Our findings indicate that 2-FBA seems to be a suitable non-oxime candidate for AChE reactivation with minimal side effects. Further toxicokinetic studies on this compound are strongly recommended to be performed before conducting the clinical trial in humans.

Keywords

Non-oxime - Diazinon - Cholinesterase activity - Pralidoxime - Organophosphate - LD50

Publication History

Received: 11 June 2022

Accepted: 29 August 2022

Article published online:
10 January 2023

• References

• 1 Marrs TC.. Organophosphate poisoning 1993; 58: 51-66
• 2 Dunn MA, Sidell FR.. Progress in Medical Defense Against Nerve Agents. 1989
• 3 Watson A, Opresko D, Young RA, Hauschild V, King J, Bakshi K.. Organophosphate Nerve Agents 2015; 87-109
• 4 Pita R, Domingo J.. The Use of Chemical Weapons in the Syrian Conflict 2014; 2: 391-402
• 5 Achilles J. Pappano. Cholinoceptor-activating and cholinesteraseinhibiting drugs. Basic Clin. Pharmacol. 14th ed.McGraw-Hill Education; 2017: pp 121–137
• 6 Stefanidou M, Athanaselis S, Spiliopoulou H.. Butyrylcholinesterase: biomarker for exposure to organophosphorus insecticides 2009; 39: 57-60
• 7 Eddleston M, Buckley NA, Eyer P, Dawson AH.. Management of acute organophosphorus pesticide poisoning. 2008
• 8 Millard CB, Kryger G, Ordentlich A, Greenblatt HM, Harel M, Raves ML. et al. Crystal structures of aged phosphonylated acetylcholinesterase: Nerve agent reaction products at the atomic level 1999; 38: 7032-7039
• 9 John H, van der Schans MJ, Koller M, Spruit HET, Worek F, Thiermann H. et al. Fatal sarin poisoning in Syria 2013: forensic verification within an international laboratory network. 2018
• 10 Szegletes T, Mallender WD, Thomas PJ, Rosenberry TL.. Substrate binding to the peripheral site of acetylcholinesterase initiates enzymatic catalysis. Substrate inhibition arises as a secondary effect 1999; 38: 122-133
• 11 Yuantari MGC, Cornelis AM, Van Gestel NM, Van S, Widianarko B, Sunoko HR. et al. Knowledge, attitude, and practice of Indonesian farmers regarding the use of personal protective equipment against pesticide exposure 2015; 187: 142
• 12 Gorecki L, Korabecny J, Musilek K, Nepovimova E, Malinak D, Kucera T. et al. Progress in acetylcholinesterase reactivators and in the treatment of organophosphorus intoxication: a patent review (2006–2016) 2017; 27: 971-985
• 13 Worek F, Thiermann H.. The value of novel oximes for treatment of poisoning by organophosphorus compounds 2013; 139: 249-259
• 14 de Koning MC, Horn G, Worek F, van Grol M.. Discovery of a potent non-oxime reactivator of nerve agent inhibited human acetylcholinesterase. 2018
• 15 Masson P.. Novel approaches in prophylaxis/pretreatment and treatment of organophosphorus poisoning. 2016
• 16 Katz FS, Pecic S, Tran TH, Trakht I, Schneider L, Zhu Z. et al. Discovery of New Classes of Compounds that Reactivate Acetylcholinesterase Inhibited by Organophosphates. 2015
• 17 Ellman GL, Courtney KD, Andres V, Featherstone RM.. A new and rapid colorimetric determination of acetylcholinesterase activity. 1961
• 18 He F.. Bradford Protein Assay 2011; 1: e45
• 19 Bajda M, Wiȩckowska A, Hebda M, Guzior N, Sotriffer CA, Malawska B.. Structure-Based Search for New Inhibitors of Cholinesterases 2013; 14: 5608-5632
• 20 Herkert NM, Freude G, Kunz U, Thiermann H, Worek F.. Comparative kinetics of organophosphates and oximes with erythrocyte, muscle and brain acetylcholinesterase 2012; 209: 173-178
• 21 Čolović MB, Krstić DZ, Lazarević-Pašti TD, Bondžić AM, Vasić VM.. Acetylcholinesterase Inhibitors: Pharmacology and Toxicology 2013; 11: 315
• 22 Worek F, Thiermann Horst, Wille T.. Organophosphorus compounds and oximes: a critical review 2020; 94: 2275-2292
• 23 De Blaquiere GE, Waters L, Blain PG, Williams FM.. Electrophysiological and biochemical effects of single and multiple doses of the organophosphate diazinon in the mouse. 2000
• 24 Montz WE, Kirkpatrick RL.. Temporal patterns of brain cholinesterase activities of white-footed mice (Peromyscus leucopus) following dosing with diazinon or parathion 1985; 14: 19-24
• 25 Aygun D, Doganay Z, Altintop L, Guven H, Onar M, Deniz T. et al. Serum acetylcholinesterase and prognosis of acute organophosphate poisoning 2002; 40: 903-910
• 26 Nouira S, Abroug F, Elatrous S, Boujdaria R, Bouchoucha S.. Prognostic value of serum cholinesterase in organophosphate poisoning 1994; 106: 1811-1814
• 27 Shih T-M, Skovira JW, O’Donnell JC, McDonough JH.. Evaluation of nine oximes on in vivo reactivation of blood, brain, and tissue cholinesterase activity inhibited by organophosphorus nerve agents at lethal dose 2009; 19: 386-400
• 28 dos Santos AA, dos Santos DB, Ribeiro RP, Colle D, Peres KC, Hermes J. et al. Effects of K074 and pralidoxime on antioxidant and acetylcholinesterase response in malathion-poisoned mice 2011; 32: 888-895