Novel arylcarbamate-N-acylhydrazones derivatives as promising BuChE inhibitors: Design, synthesis, molecular modeling and biological evaluation

https://doi.org/10.1016/j.bmc.2020.115991Get rights and content

Highlights

  • Design arylcarbamate-N-acylhydrazones derivatives were BuChE selective inhibitors.

  • The compounds were synthesized using low-cost and simple methodologies.

  • Compounds 10a-d are potent BuChE inhibitors, with IC50 values of 0.07–0.56 µM.

  • Best inhibitor 10c is non-competitive type of BuChE.

  • Docking studies confirmed important interactions of 10c with hBuChE active site.

Abstract

A novel series of arylcarbamate-N-acylhydrazones derivatives have been designed and synthesized as potential anti-cholinesterase agents. In vitro studies revealed that these compounds demonstrated selective for butyrylcholinesterase (BuChE) with potent inhibitory activity. The compounds 10a-d, 12b and 12d were the most potent BuChE inhibitors with IC50 values of 0.07–2.07 µM, highlighting the compound 10c (IC50 = 0.07 µM) which showed inhibitory activity 50 times greater than the reference drug donepezil (IC50 = 3.54 µM). The activity data indicates that the position of the carbamate group in the aromatic ring has a greater influence on the inhibitory activity of the derivatives. The enzyme kinetics studies indicate that the compound 10c has a non-competitive inhibition against BuChE with Ki value of 0.097 mM. Molecular modeling studies corroborated the in vitro inhibitory mode of interaction and show that compound 10c is stabilized into hBuChE by strong hydrogen bond interaction with Tyr128, π-π stacking interaction with Trp82 and CH⋯O interactions with His438, Gly121 and Glu197. Based on these data, compound 10c was identified as low-cost promising candidate for a drug prototype for AD treatment.

Introduction

Cholinesterase inhibitors (ChEI) are a group of drugs that block the normal hydrolysis (cholinesterase-induced) of the neurotransmitter acetylcholine (ACh) in acetate and choline, and subsequent increasing the levels and duration of the acetylcholine in the central nervous system.1, 2, 3, 4 These inhibitor act in the synaptic cleft, inhibiting the cholinesterase enzymes.3, 4 ChEI are the most promising agents developed until today, for the symptomatic treatment of diseases related to cholinergic systems, such as: Alzheimer’s disease (AD),3, 4 glaucoma5 and myasthenia gravis6. The use of these therapeutic agents has improved the cognitive functions of patients affected by AD.3, 4

AD is a multifactorial neurodegenerative disorder characterized by cognitive impairment, mainly affects elderly.7, 8 Due to the aging of the world's populations is a public health problem, with a great human, social and economic burden.9 According World Alzheimer Report 2019, 50 million people are living with dementia worldwide, and the projection for 2050 is estimated to increase to more than 152 million.10, 11 Thus, enormous material and financial resources are devoted into treatment of AD, with the current annual cost of dementia is estimated at US $1trillion, and the projection for 2030 a figure set to double.10

Depending on the AD stage, there is a decline in acetylcholinesterase (AChE) levels in the brain and a progressive increase of butyrylcholinesterase (BuChE) which becomes responsible for the hydrolysis of ACh.12, 13, 14 According to amyloid hypothesis, fibrillar β-amyloid disorder is a neuropathology of AD that may be related to neural decrease.15, 16 The increase BuChE levels may be associated with the formation of these toxic fibrillar β-amyloid characteristic of AD, however, a mechanism for this process has not been elucidated.17, 18

AChE knockout mice studies have shown the importance role of BuChE in the nervous system as coregulator of ACh. BuChE is able to compensate for the lack of AChE, allowing the continued regulation of cholinergic neurotransmission.19 Moreover, BuChE knockout mice studies demonstrated the fibrillar β-amyloid plaques deposition in subcortical regions of the brain was reduced with BuChE deficiency. In this way, the decrease BuChE levels contributed to increase learning capacity and lowered the vulnerability β-amyloid toxicity.20, 21, 22, 23 Therefore, innumerous studies have been performed to development of selective BuChE inhibitors.24, 25, 26, 27, 28 Since the reduction of BuChE levels can be an important cholinergic therapeutic approach aimed at controlling the development of AD pathology.29

The anticholinesterasic property of carbamates has been known for decades. The first carbamate used clinically as ChEI in the treatment of AD was physostigmine. However, its use has been discontinued due to its high doses and side effects.16 Other carbamates such as neostigmine and pyridostigmine are used to treat myasthenia gravis, and neostigmine is also used in the glaucoma treatment. In addition, rivastigmine is one of the four drugs approved by the Food and Drug Administration (FDA) for the treatment of AD.30, 31 In the discovery of new bioactive compounds, the N-acylhydrazone moiety is considered a privileged structure, capable of acting as a pharmacophore or auxophore subunit in different pharmaceutical classes.32 In the last years, compounds containing the N-acylhydrazone portion have been studied as ChEI.33, 34, 35, 36

Based on these considerations, a series of arylcarbamate-N-acylhydrazone derivatives have been designed, synthesized and evaluated in vitro which were expected to inhibit cholinesterases. These compounds were designed by combining arylcarbamate and 3,4-dimethoxybenzyl scaffolds linked with N-acylhydrazone moiety (see Fig 1).

Section snippets

Synthesis

The target compounds (10a-12d) were synthesized via the route outlined in Scheme 1. Carboxylic acids 1a-c were used as starting materials to prepare carbohydrazide intermediates 3a-d. In the route A, carboxylic acids 1a-1c were reacted with SOCl2 at reflux of ethanol for 12 h37 to provide ester derivatives 2a-c (72 – 89% yield) by one pot esterification reaction. To obtain methyl 3,4-dimethoxybenzoperoxoate 2d, the carboxylic acid 1c was reacted with Me2SO4 in the presence of K2CO3 at reflux of

Conclusions

A series of new arylcarbamate-N-acylhidrazones derivatives (10a-12d) was designed and synthesized using low-cost starting materials and simple methodologies. Enzymatic inhibition studies show that the compounds 10a-d, 12b and 12d were selective and potent BuChE inhibitors, with IC50 values of 0.07–2.07 µM. The greater inhibitory potency towards BuChE is showed by compound 10c (Ki 0.097 mM), with orto-carbamate-N-acylhydrazone groups in the ring A and and 3-methoxy-4-hydroxybenzyl scaffold

Chemicals

Starting materials and reagents were purchased from Sigma-Aldrich and Acros. For column chromatography, silica gel 60, 230–400 mesh (Merck) was used. Nuclear magnetic resonance (NMR) spectra were acquired with Varian Mercury Plus BB 300 MHz and Bruker Avance III HD 300 and 500 MHz. The spectra were recorded in 20 mg cm-3 solutions of DMSO‑d6, with a probe temperature of ca. 300 K and tetramethylsilane (TMS) as reference. High-resolution mass spectrometry (HRMS) analysis was performed in a

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgements

We would like to thank Fundação Araucária (002/17) and CNPQ (404455/2016-6) for the financial support of this research, and CAPES for a scholarship (Yamazaki, D. A. S.; Baréa, P.; Reis, E. C. and Rozada, A. M. F.) and to Complexo de Centrais de Apoio à Pesquisa of Universidade Estadual de Maringá (COMCAP-UEM) for the facilities.

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