Discovery of Potent Cholinesterase Inhibition-Based Multi-Target-Directed Lead Compounds for Synaptoprotection in Alzheimer’s Disease

Abstract

Drug development efforts that focused on single targets failed to provide effective treatment for Alzheimer’s disease (AD). Therefore, we designed cholinesterase inhibition (ChEI)-based multi-target-directed ligands (MTDLs) to simultaneously target AD-related receptors. We built a library of 70 compounds, sequentially screened for ChEI, and determined σ1R, σ2R, NMDAR-GluN2B binding affinities, and P2X7R antagonistic activities. Nine fulfilled in silico drug-likeness criteria and did not display toxicity in three cell lines. Seven displayed cytoprotective activity in two stress-induced cellular models. Compared to donepezil, six showed equal/better synaptic protection in a zebrafish model of acute amyloidosis-induced synaptic degeneration. Two P2X7R antagonists alleviated the activation state of microglia in vivo. Permeability studies were performed, and four did not inhibit CYP450 3A4, 2D6, and 2C9. Therefore, four ChEI-based lead MTDLs are promising drug candidates for synaptic integrity protection and could serve as disease-modifying AD treatment. Our study also proposes zebrafish as a useful preclinical tool for drug discovery and development.

Potential bioactive compounds and mechanisms of Fibraurea recisa Pierre for the treatment of Alzheimer’s disease analyzed by network pharmacology and molecular docking prediction

“…Analysis results of their molecular structures indicated that the modification of these two alkaloids could be exerted at multiple sites, which may probably improve their bioavailability and biological activity ( Shan et al, 2011 ; Yan B. et al, 2017 ; Liang et al, 2021 ; Castellano et al, 2022 ). Notably, the rational structural modifications of AChE inhibitors were still the primary route to the discovery of novel One-molecule-multi-target anti-AD drugs at this stage ( Rossi et al, 2021 ; Sang et al, 2022 ; Turgutalp et al, 2022 ). Since these two alkaloids’ high potential in vitro acetylcholinesterase inhibitory activity had been verified by other researchers ( Jia et al, 2021 ; Tang et al, 2021 ; Miao et al, 2022 ).…”

Zebrafish as an Experimental and Preclinical Model for Alzheimer’s Disease

“…Given the multifactorial nature of AD, a drug treatment approach that targets multiple causative agents (existing or yet-to-be-identified) simultaneously will be a rewarding route. We recently used our acute amyloidosis model in zebrafish to successfully evaluate potent cholinesterase (ChE) inhibition-based multitarget-directed ligands for their efficacy and mechanism of action on synaptic integrity . We designed a structural scaffold targeting AD-related receptors with ChE background inhibition that not only yields a symptomatic effect but also could provide DMT through neuroprotection.…”

“…We performed extensive cytotoxicity screening and obtained few promising compounds that were tested for their in vitro cytoprotective efficacies. These studies displayed equal/better synaptic protection compared to benchmark drug donepezil in our zebrafish model of acute amyloidosis-induced synaptic degeneration . This in vivo study allowed us to increase the number of candidates for further drug discovery studies and provided a pipeline that could be successfully used to develop first-in-class drug candidates by performing a phenotypic screening of a drug library that targets key AD-related proteins (Figure ).…”

“…We recently used our acute amyloidosis model in zebrafish to successfully evaluate potent cholinesterase (ChE) inhibition-based multitarget-directed ligands for their efficacy and mechanism of action on synaptic integrity. 6 We designed a structural scaffold targeting AD-related receptors with ChE background inhibition that not only yields a symptomatic effect but also could provide DMT through neuroprotection. We performed extensive cytotoxicity screening and obtained few promising compounds that were tested for their in vitro cytoprotective efficacies.…”

Ngfr suppresses Lcn2/Slc22a17 signaling, induces neurogenesis and reduces amyloid pathology in the hippocampus of APP/PS1dE9 mouse

“…We previously generated an adult AD zebrafish model and found that a complex set of cellular crosstalk leads to production of more neurons despite the disease ongoing pathology 9–12 . This model has strong parallelism to human AD brains in terms of molecular programs affected by AD pathology 13 , served as a pre-clinical tool for drug screening 14–16 and helped identify genes associated with AD in humans 17,18 . The extraordinary regenerative ability of astroglia in zebrafish in AD-like scenarios rely on set of molecular mechanisms, one of which is the pro-neurogenic activity through the expression of nerve growth factor receptor ( ngfra ) 9,19 .…”