Novel Hybrid Acetylcholinesterase Inhibitors Induce Differentiation and Neuritogenesis in Neuronal Cells in vitro Through Activation of the AKT Pathway

Abstract

Background: Alzheimer’s disease (AD) is characterized by a progressive loss of episodic memory associated with amyloid-β peptide aggregation and the abnormal phosphorylation of the tau protein, leading to the loss of cholinergic function. Acetylcholinesterase (AChE) inhibitors are the main class of drugs used in AD therapy. Objective: The aim of the current study was to evaluate the potential of two tacrine-donepezil hybrid molecules (TA8Amino and TAHB3), which are AChE inhibitors, to induce neurodifferentiation and neuritogenesis in SH-SY5Y cells. Methods: The experiments were carried out to characterize neurodifferentiation, cellular changes related to responses to oxidative stress and pathways of cell survival in response to drug treatments. Results: The results indicated that the compounds did not present cytotoxic effects in SH-SY5Y or HepG2 cells. TA8Amino and TAHB3 induced neurodifferentiation and neuritogenesis in SH-SY5Y cells. These cells showed increased levels of intracellular and mitochondrial reactive oxygen species; the induction of oxidative stress was also demonstrated by an increase in SOD1 expression in TA8Amino and TAHB3-treated cells. Cells treated with the compounds showed an increase in PTEN(Ser380/Thr382/383) and AKT(Ser473) expression, suggesting the involvement of the AKT pathway. Conclusion: Our results demonstrated that TA8Amino and TAHB3 present advantages as potential drugs for AD therapy and that they are capable of inducing neurodifferentiation and neuritogenesis.

Novel Dual AChE and ROCK2 Inhibitor Induces Neurogenesis via PTEN/AKT Pathway in Alzheimer’s Disease Model

“…In previous studies, it was demonstrated that the donepezil–tacrine hybrid compound (TA8Amino) showed a potent allosteric modulator of AChE and induced promising effects in terms of neuroprotection and neurodifferentiation [ 32 , 38 ]. These studies highlight that TA8Amino demonstrated low toxicity and a potent inhibitory effect on AChE at lower concentrations than donepezil and tacrine.…”

“…For AChEIs-induced neuronal differentiation assays, the same protocol was performed (except for the addition of RA). The analysis of the percentage of neuronal differentiation and neurite outgrowth was performed according to a previous protocol [ 38 ]. Briefly, a total of 20 photographs (20× magnification) were taken of each group treatment using an inverted phase microscope (Zeiss, Jena, Germany) to quantify the percentage of differentiated cells and neurite size; the ImageJ software (Fiji, image processing package) and the Simple Neurite Tracer plugin were used for these measurements.…”

Neuroprotective Effects of Cholinesterase Inhibitors: Current Scenario in Therapies for Alzheimer’s Disease and Future Perspectives

“…Inhibition of PTEN in AD mouse models is associated with several consequences, such as reduced expression of Bax protein, decreased apoptosis, lower levels of DNA fragmentation, reduced endoplasmic reticulum stress [ 130 ], and rescue of normal synaptic function and cognition [ 131 ]. Noteworthy, reduced activation of PTEN and increased AKT phosphorylation might be involved in neuritogenesis and neurodifferentiation [ 84 ]. Accordingly, targeting PTEN to induce AKT activation might be a potential approach to be investigated in AD therapy ( Fig.…”

Caracterização dos efeitos de neuroproteção e neurodiferenciação de novos compostos inibidores de acetilcolinesterase em células PC12 diferenciadas em neurônios

“…Foram desenvolvidos então dois compostos (TA8Amino e TAHB3) que demonstraram eficiência superior aos compostos originais com relação à inibição da AChE (Chierrito et al, 2017;Moreira et al, 2020) Figura 2: Representação das estruturas moleculares dos compostos AChEIs com seus respectivos valores de 50% da ação inibitória máxima (IC50). Compostos originais donepezila e tacrina e os novos compostos híbridos TAHB3 e TA8Amino (dados extraídos de Chierrito, et al, 2017).…”

“…Os resultados já obtidos demonstraram que, por meio da ativação da via PI3K/Akt (do inglês, phosphoinositide 3-kinase e Protein Kinase B), os compostos estimularam a neurodiferenciação e neuritogênese nas células SH-SY5Y; os compostos também demonstraram baixos níveis de citotoxicidade e não induziram alterações na viabilidade celular nas concentrações próximas ao IC50 de inibição da AChE. Também foi observado um efeito neuroprotetor em resposta a danos induzidos pelo peptídeo Aβ (Moreira, 2019;Moreira et al, 2020). Esses dados preliminares demonstram o potencial terapêutico dos compostos TA8Amino e TAHB3 para DA, e ressaltam a importância da continuação de estudos com essas moléculas em diferentes modelos e linhagens celulares para comprovar a capacidade indutora de diferenciação e de efeitos neuroprotetores 1.5.…”