Research Paper Volume 17, Issue 3 pp 822—850
Differential senolytic inhibition of normal versus Aβ-associated cholinesterases: implications in aging and Alzheimer’s disease
- 1 Department of Medical Neuroscience, Dalhousie University, Halifax, Nova Scotia B3H 4R2, Canada
- 2 Department of Medicine (Geriatric Medicine and Neurology), Dalhousie University, Halifax, Nova Scotia B3H 2E1 Canada
Received: December 4, 2024 Accepted: March 13, 2025 Published: March 29, 2025
https://doi.org/10.18632/aging.206227How to Cite
Copyright: © 2025 Darvesh et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
Cellular senescence is a hallmark of aging and the age-related condition, Alzheimer’s disease (AD). How senescence contributes to cholinergic and neuropathologic changes in AD remains uncertain. Furthermore, little is known about the relationship between senescence and cholinesterases (ChEs). Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) are important in neurotransmission, cell cycle regulation, and AD amyloid-β (Aβ) pathology. Senolytic agents have shown therapeutic promise in AD models. Therefore, we evaluated in vitro and in silico activity of senolytics, dasatinib (1), nintedanib (2), fisetin (3), quercetin (4), GW2580 (5), and nootropic, meclofenoxate hydrochloride (6), toward AChE and BChE. As ChEs associated with AD pathology have altered biochemical properties, we also evaluated agents 1-6 in AD brain tissues. Enzyme kinetics showed agents 1, 3, 4, and 6 inhibited both ChEs, while 2 and 5 inhibited only AChE. Histochemistry showed inhibition of Aβ plaque-associated ChEs (1 and 2: both ChEs; 5: BChE; 6: AChE), but not normal neural-associated ChEs. Modeling studies showed 1-6 interacted with the same five binding locations of both ChEs, some of which may be allosteric sites. These agents may exert their beneficial effects, in part, by inhibiting ChEs associated with AD pathology and provide new avenues for development of next-generation inhibitors targeting pathology-associated ChEs.
Abbreviations
Aβ: amyloid-β; ABP: acyl binding pocket; AChE: acetylcholinesterase; AD: Alzheimer’s disease; ATChI: acetylthiocholine iodide; BChE: butyrylcholinesterase; Bibfl120: nintedanib; BTChI: butyrylthiocholine iodide; CAIP: cholinergic anti-inflammatory pathway; CAS: catalytic active site; ChAT: choline acetyltransferase; ChE: cholinesterase; ChEI: cholinesterase inhibitor; DAB: 3,3’-diaminobenzidine tetrahydrochloride; dH2O: distilled water; DTNB: 5,5-dithio-bis-(2-nitrobenzoic acid); Ki: inhibition constant; H2O2: hydrogen peroxide; IL-1β: interleukin 1β; IL-6: interleukin 6; KR: Karnovsky-Roots; mAChR: muscarinic acetylcholine receptor; MOE: Molecular Operating Environment; nAChR: nicotinic acetylcholine receptor; OAH: oxyanion hole; PAS: peripheral anionic site; PB: phosphate buffer; PCS: π-cationic site; PDB: Protein Databank; SAMP8: senescence-accelerated prone 8 mouse; SASP: senescence-associated secretory phenotype; TNB: 5-thio-2-nitrobenzoic acid; TNFα: tumor necrosis factor α.