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Neostigmine is a competitive reversible inhibitor of acetylcholinesterase. When the drug interacts with the serine of the enzyme's active site, a nucleophilic substitution reaction takes place where the drug loses a phenol group leading to the formation of a stable intermediate. However, serine then gets capped by a stable urethane group. Hydrolysis then occurs preventing "aging" from happening. The serine will then return to its normal form so this inhibition is reversible.

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