Organophosphorus compounds (OPs) are used as pesticides, e.g. parathion, which is converted in the body to paraoxon, and chemical warfare nerve agents, such as sarin, soman, cyclosarin, VX, and tabun. Even small amounts of OP exposure can be fatal, depending on the toxicity of the compound. Great stocks of highly toxic chemical warfare nerve agents exit around the world and are considered a serious threat to national security and international stability. OPs exert their toxicity by covalent irreversible inhibition of acetylcholinesterase (AChE) that prevents the enzyme from hydrolyzing acetylcholine (ACh), a neurotransmitter in the central and peripheral nervous systems (CNS and PNS). Therefore, ACh accumulates in the cholinergic synapses throughout the body, which results in overstimulation of the ACh receptors. Removal of the phosphyl moiety from the OP-bound AChE active site has been a promising method to restore AChE’s catalytic activity. However, a secondary process called aging also occurs in the OP-AChE complex. Once aging occurs, currently available oximes are ineffective in removing the phosphyl moiety from the enzyme’s active site, and hence are ineffective as antidotes against the aged enzyme. Several families of alkylating and acylating agents including several classes of agents that combine alkylating moieties with known active site or peripheral cite (PAS) binding motifs were synthesized and evaluated. The general aim of the research was that successful alkylation or acylation of the phosphonate monoanion of aged AChE would produce neutral phosphyl complexes that would either spontaneously reactivate or would be reactivatable in the presence of oxime antidotes. Methoxylamine analogs of the oxime antidote 2-PAM were synthesized with the aim that methyl transfer to the aged AChE adduct would produce a neutral phosphyl AChE adduct simultaneously with 2-PAM in situ, and subsequent 2-PAM nucleophilic attack would reactivate the newly formed neutral phosphyl-AChE adduct. However, none of these 2-PAM analogs resurrected the activity of aged AChE. Another strategy for resurrecting the activity of aged AChE utilizes N-methylpyridiniums that are substituted at the 2-position with a beta-lactam moiety. For these compounds, opening of the electrophilic beta-lactam unmasks a nucleophilic amidine function which could putatively attack at phosphorus to expel the free enzyme. For this class of agents, only the active site directed compound that possessed the 5-CF₃ substituent showed possible resurrection of the activity of aged AChE, though activities in both the control and treated samples were low. Methyl transfers are common in Nature, and the natural transfer agent is S-adenosylmethionine, a sulfonium methyl donor. Consequently, the array of sulfonium compounds were evaluated on the expectation that they would bind to the AChE active site and transfer a methyl group to the phosphonate monoanion of the aged enzyme. Though high-affinity binding was noted for these compounds, none of these resurrected the activity of the aged AChE complex. Finally, several selected agents were evaluated on reactivating the initial OP-AChE complex before aging has occurred. It was observed that degraded samples of selected inhibitors are capable of reactivating initial complexes of sarin and soman inhibited AChE at low concentration that is an important character of efficient reactivators. However, the structure of reactivator is still unknown. Two major challenges still face researchers in the quest to design effective medicinal agents for counteracting poisoning by AChE-inhibiting nerve agents. The first is that there is no universal oxime antidote. Oximes that are effective against certain nerve agents are ineffective against others. The second is that, despite extensive efforts that span two generations, aged phosphyl-AChE adducts have never been reactivated. However, given the powerful tools of modern structural biology, medicinal chemistry and molecular biology, there is still hope that these considerable challenges can be met.