Straightforward, Economical Procedures for Microscale Ellman’s Test for Cholinesterase Inhibition and Reactivation

Cavalcante, Samir F. de A.; Kitagawa, Daniel A. S.; Rodrigues, Rafael B.; Cardozo, Monique; Paula, Reuel L. de; Correa, Ana Beatriz de A.; Simas, Alessandro B. C.

doi:10.21577/0100-4042.20170278

Cited by 5 publications

(11 citation statements)

References 27 publications

(57 reference statements)

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“…The results of Ellman’s tests [ 40 , 41 ] show that isatin-O can reactivate Ee AChE/POX at 10 and 100 μmol/L. At 10 μmol/L, it reached around 8% reactivation in the initial 10 min.…”

Section: Resultsmentioning

confidence: 99%

“…The Ellman method was employed on the biochemical evaluation of AChE activity, being a classic method of enzymatic evaluation [ 40 ]. In this study, the method was adapted to microscale to work with small doses of OPs as well as to reduce the risks of accidental intoxication [ 41 ]. In addition, scale reduction also provided significant savings in materials and reagents.…”

Section: Methodsmentioning

confidence: 99%

“…For the in vitro studies, we used an Ellman’s [ 40 ] modified method [ 41 ]. Since its first publication, this is the most common and relevant assay for investigating AChE activity [ 42 , 43 ].…”

Section: Introductionmentioning

confidence: 99%

“…Since its first publication, this is the most common and relevant assay for investigating AChE activity [ 42 , 43 ]. The method was developed in the early 1960s [ 40 ] and it is still in use today, with some further improving modifications [ 41 , 44 ]. Despite some limitations eventually found, such as the lack of low detection limit and frequent background perturbations, this method has been extensively used [ 42 ] due to its simplicity, accuracy, low cost and the large number of validated and published assays.…”

Section: Introductionmentioning

confidence: 99%

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Molecular Modeling and In Vitro Studies of a Neutral Oxime as a Potential Reactivator for Acetylcholinesterase Inhibited by Paraoxon

et al. 2018

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The present work aimed to compare the small, neutral and monoaromatic oxime, isatin-3-oxime (isatin-O), to the commercial ones, pralidoxime (2-PAM) and obidoxime, in a search for a new potential reactivator for acetylcholinesterase (AChE) inhibited by the pesticide paraoxon (AChE/POX) as well as a novel potential scaffold for further synthetic modifications. The multicriteria decision methods (MCDM) allowed the identification of the best docking poses of those molecules inside AChE/POX for further molecular dynamic (MD) studies, while Ellman’s modified method enabled in vitro inhibition and reactivation assays. In corroboration with the theoretical studies, our experimental results showed that isatin-O have a reactivation potential capable of overcoming 2-PAM at the initial moments of the assay. Despite not achieving better results than obidoxime, this molecule is promising for being an active neutral oxime with capacity of crossing the blood–brain barrier (BBB), to reactivate AChE/POX inside the central and peripheral nervous systems. Moreover, the fact that isatin-O can also act as anticonvulsant makes this molecule a possible multipotent reactivator. Besides, the MCDM method showed to be an accurate method for the selection of the best docking poses generated in the docking studies.

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“…The results of Ellman’s tests [ 40 , 41 ] show that isatin-O can reactivate Ee AChE/POX at 10 and 100 μmol/L. At 10 μmol/L, it reached around 8% reactivation in the initial 10 min.…”

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Molecular Modeling and In Vitro Studies of a Neutral Oxime as a Potential Reactivator for Acetylcholinesterase Inhibited by Paraoxon

et al. 2018

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“…During experiments, all solutions were kept at 0 °C. DMSO did not affect measurements in our conditions [60].…”

Section: Methodsmentioning

confidence: 99%

In Vitro Evaluation of Neutral Aryloximes as Reactivators for Electrophorus eel Acetylcholinesterase Inhibited by Paraoxon

et al. 2019

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Casualties caused by organophosphorus pesticides are a burden for health systems in developing and poor countries. Such compounds are potent acetylcholinesterase irreversible inhibitors, and share the toxic profile with nerve agents. Pyridinium oximes are the only clinically available antidotes against poisoning by these substances, but their poor penetration into the blood-brain barrier hampers the efficient enzyme reactivation at the central nervous system. In searching for structural factors that may be explored in future SAR studies, we evaluated neutral aryloximes as reactivators for paraoxon-inhibited Electrophorus eel acetylcholinesterase. Our findings may result into lead compounds, useful for development of more active compounds for emergencies and supportive care.

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Are the current commercially available oximes capable of reactivating acetylcholinesterase inhibited by the nerve agents of the A-series?

et al. 2022

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Straightforward, Economical Procedures for Microscale Ellman’s Test for Cholinesterase Inhibition and Reactivation

Cited by 5 publications

References 27 publications

Molecular Modeling and In Vitro Studies of a Neutral Oxime as a Potential Reactivator for Acetylcholinesterase Inhibited by Paraoxon

Molecular Modeling and In Vitro Studies of a Neutral Oxime as a Potential Reactivator for Acetylcholinesterase Inhibited by Paraoxon

In Vitro Evaluation of Neutral Aryloximes as Reactivators for Electrophorus eel Acetylcholinesterase Inhibited by Paraoxon

Are the current commercially available oximes capable of reactivating acetylcholinesterase inhibited by the nerve agents of the A-series?

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