Perspective

Drug Candidates for the Treatment of Alzheimer’s Disease:

New Findings from 2021 and 2022
Sujatha L. Motebennur 1 , Belakatte P. Nandeshwarappa 1 and Manjunatha S. Katagi 2, *

1 Department of Studies in Chemistry, Davangere University, Davangere 577 007, India

2 Department of Pharmaceutical Chemistry, Bapuji Pharmacy College, Davangere 577 004, India
* Correspondence: [email protected]; Tel.: +91-98-8649-9160

Abstract: Alzheimer’s disease (AD), an ongoing neurodegenerative disorder among the elderly, is
signalized by amnesia, progressive deficiency in cognitive roles, and behavioral deformity. Over the
last ten years, its pathogenesis still remains unclear despite several efforts from various researchers
across the globe. There are certain factors that seem to be involved in the progression of the disease
such as the accumulation of β-amyloid, oxidative stress, the hyperphosphorylation of tau protein, and
a deficit of acetylcholine (ACh). Ongoing therapeutics are mainly based on the cholinergic hypothesis,
which suggests that the decrease in the ACh levels leads to the loss of memory. Therefore, increasing
the cholinergic function seems to be beneficial. Acetylcholinesterase inhibitors (AChEIs) inhibit the
enzyme by avoiding the cleavage of acetylcholine (ACh) and increasing the neurotransmitter acetyl-
choline (ACh) levels in the brain areas. Thus, the cholinergic deficit is the root cause of Alzheimer’s
disease (AD). Currently, drugs such as tacrine, donepezil, rivastigmine, and galantamine have been
launched on the market for a cholinergic approach to AD to increase neurotransmission at cholinergic
synapses in the brain and to improve cognition. These commercialized medicines only provide
supportive care, and there is a loss of medicinal strength over time. Therefore, there is a demand
for investigating a novel molecule that overcomes the drawbacks of commercially available drugs.
Therefore, butyrylcholinesterase (BChE), amyloid-β (Aβ), β-secretase-1 (BACE), metals Cu(II), Zn(II),
or Fe(II), antioxidant properties, and the free radical scavenging capacity have been primarily tar-
geted in the preceding five years along with targeting the AChE enzyme. A desired, well-established
Citation: Motebennur, S.L.;
pharmacological profile with a number of hybrid molecules incorporating substructures within a
Nandeshwarappa, B.P.; Katagi, M.S.
single scaffold has been investigated. From distinct chemical categories such as acridine, quinoline,
Drug Candidates for the Treatment of
carbamate, huperzine, and other heterocyclic analogs, the main substructures used in developing
Alzheimer’s Disease: New Findings
from 2021 and 2022. Drugs Drug
these molecules are derived. The optimization of activity through structural modifications of the
Candidates 2023, 2, 571–590. https:// prototype molecules has been followed to develop the Structure Activity Relationship (SAR), which in
doi.org/10.3390/ddc2030030 turn facilitates the development of novel molecules with expected AChE inhibitory activity together
with many more pharmacological properties. The present review outlines the current drug candidates
Academic Editor: Jean Jacques
in the advancement of these AChEIs in the last two years.
Vanden Eynde

Received: 22 March 2023 Keywords: acetylcholinesterase; Alzheimer’s disease; acetylcholine; acetylcholinesterase inhibitors;
Revised: 27 June 2023 IC50 ; molecular docking
Accepted: 7 July 2023
Published: 17 July 2023

1. Introduction
Copyright: © 2023 by the authors.
Dementia is a set of manifestations that often occur together or a condition charac-
Licensee MDPI, Basel, Switzerland. terized by a group of interrelated symptoms such as deterioration in memory, learning
This article is an open access article capacity, thinking, judgment, orientation, comprehension, calculation, language, and the
distributed under the terms and inability to carry out daily activities [1–3]. Worldwide, an average of 50 million people are
conditions of the Creative Commons living with dementia, and this estimated figure is expected to increase to 75 million in 2030
Attribution (CC BY) license (https:// and 131.5 million in 2050 due to the rise in the elderly population [4,5]. The monetary stress
creativecommons.org/licenses/by/ associated with dementia management indicates that approximately 600 billion dollars are
4.0/). spent annually to care for over 45 million individuals with dementia, which is roughly 1%

Drugs Drug Candidates 2023, 2, 571–590. https://doi.org/10.3390/ddc2030030 https://www.mdpi.com/journal/ddc

Drugs Drug Candidates2023, 2, FOR PEER REVIEW 2

stress associated with dementia management indicates that approximately 600 billion dol-
Drugs Drug Candidates 2023, 2
lars are spent annually to care for over 45 million individuals with dementia, which572 is
roughly 1% of the global gross domestic product [6,7]. With the health care system, this
expenditure is likely to increase as the population grows and more elderly individuals
live into their declining years [8,9]. Alzheimer’s disease (AD), being a form of dementia,
of the globaltogross
is expected domestic
contribute product [6,7].
significantly With
to future the health
expenses. care system,
Studies have shownthis expenditure
that provid-
is likely to increase as the population grows and more elderly
ing care for all people living with AD by 2080 could lead to an increased total individuals live into their
budget due
declining years [8,9]. Alzheimer’s disease (AD), being a form of dementia, is expected to
to prolonging the period of a patient’s stay in mild, moderate, or severe AD stages [10,11].
contribute significantly to future expenses. Studies have shown that providing care for all
AD is an incurable neurodegenerative brain disorder that leads to mental health dis-
people living with AD by 2080 could lead to an increased total budget due to prolonging
orders primarily affecting cognitive abilities, including learning, perception, and memory
the period of a patient’s stay in mild, moderate, or severe AD stages [10,11].
loss. It poses a significant burden on patients, their families, and the healthcare system
AD is an incurable neurodegenerative brain disorder that leads to mental health disor-
due to the profound disability and helplessness it entails. AD is prevalent worldwide
ders primarily affecting cognitive abilities, including learning, perception, and memory
across various demographic groups, although it is more common in older individuals.
loss. It poses a significant burden on patients, their families, and the healthcare system
Consequently, AD represents one of the greatest global health challenges for society
due to the profound disability and helplessness it entails. AD is prevalent worldwide
[12,13].
across various demographic groups, although it is more common in older individuals.
Alzheimer’s disease is characterized by three main categories of manifestations. The
Consequently, AD represents one of the greatest global health challenges for society [12,13].
first category, cognitive dysfunction, involves loss of memory, difficulties in language,
Alzheimer’s disease is characterized by three main categories of manifestations. The
and executive dysfunction.
first category, cognitive The secondary
dysfunction, involvescategory comprises
loss of memory, behavioral
difficultiesdisturbances,
in language,
and executive dysfunction. The secondary category comprises behavioral referred
such as depression, illusions, misapprehension, and anxiety, collectively to as
disturbances,
non-cognitive
such symptoms
as depression, and psychiatric
illusions, symptoms
misapprehension, and[14]. The third
anxiety, categoryreferred
collectively involvestodif-as
non-cognitive symptoms and psychiatric symptoms [14]. The third categoryshopping,
ficulties in performing daily activities such as orientation, judgment, driving, involves
and basic tasks
difficulties like dressing
in performing dailyand eatingsuch
activities independently.
as orientation,The symptoms
judgment, of Alzheimer’s
driving, shopping,
disease progress from mild to very severe, starting with memory
and basic tasks like dressing and eating independently. The symptoms of Alzheimer’s loss and eventually lead-
ing to dementia
disease progress[15].from mild to very severe, starting with memory loss and eventually
Alzheimer’s
leading to dementia disease
[15]. is associated with numerous obsessive characteristics including
inadequacy of the neurotransmitter
Alzheimer’s disease is associated ACh,withthenumerous
accumulation of beta-amyloid
obsessive (Aβ)including
characteristics plaques,
increased MAO-B enzyme activity, over-stimulus of the N-methyl-D-aspartate
inadequacy of the neurotransmitter ACh, the accumulation of beta-amyloid (Aβ) plaques, receptor,
changes inMAO-B
increased the homeostasis of biometals,
enzyme activity, oxidativeofstress,
over-stimulus and hyperphosphorylation
the N-methyl-D-aspartate of
receptor,
tau protein
changes [16].
in the homeostasis of biometals, oxidative stress, and hyperphosphorylation of tau
The[16].
protein current management of AD focuses on inhibiting cholinesterase, thereby improv-
ing the
Theconcentration
current management of ACh in ofthe
ADsynaptic
focuses on cleft. One of the
inhibiting oldest and best
cholinesterase, therebyunderstood
improv-
neurotransmitters
ing the concentration in the mammalian
of ACh central nervous
in the synaptic cleft. Onesystem
of the (CNS)
oldest is ACh.
and bestItunderstood
is localized
in various parts of in
neurotransmitters thetheCNS and is synthesized
mammalian in andsystem
central nervous released fromischolinergic
(CNS) neurons.
ACh. It is localized
Cholinergic
in neurons
various parts of the projecting
CNS andfrom basal forebrain
is synthesized in andnuclei playfrom
released a crucial role in cognitive
cholinergic neurons.
processes such
Cholinergic as learning
neurons and from
projecting memory.
basalCholinergic neurons
forebrain nuclei playina the corpus-striatum
crucial role in cognitive are
involved in
processes motor
such functions.
as learning andCholine
memory. O-acetyltransferase
Cholinergic neurons (ChAT) facilitates
in the the synthesis
corpus-striatum are
of ACh from
involved choline
in motor and acetyl
functions. CoA within
Choline presynaptic cholinergic
O-acetyltransferase neurons
(ChAT) facilitates the[17] (Scheme
synthesis of
ACh
1). from choline and acetyl CoA within presynaptic cholinergic neurons [17] (Scheme 1).

O CH3
CH3 H3 C O CH3
H3 C OH N HS CoA
N CoA
H3C S Coenyme A
CH3 O
CH3 Acetyl CoA Acetylcholine
Choline

Scheme 1. Synthesis of Acetylcholine.

According
According totothethe
International Union
International of Biochemistry
Union and Molecular
of Biochemistry Biology (IUBMB),
and Molecular Biology
AChE
(IUBMB),(3.1.1.7)
AChEbelongs
(3.1.1.7)tobelongs
the class
toof
theserine
class hydrolases that act on
of serine hydrolases ester
that actbonds of bonds
on ester esters
of
of carboxylic acids. Theacids.
esters of carboxylic significant characteristic
The significant of the AChE
characteristic structure
of the AChE is its widening
structure is its
at the base, forming a wide gorge that extends about half a molecule deep [18,19]. The
enzyme structure of AChE has been elucidated through X-ray crystallographic studies
using Torpedo californica (PDB ID: 1ACJ and 1ACL). These studies revealed that AChE
possesses two main binding sites: the catalytic active site (CAS) and the peripheral anionic
site (PAS) connected to the gorge within the enzyme’s active site [20–23]. Amino acids
 widening at the base, forming a wide gorge that extends about half a molecule deep
[18,19]. The enzyme structure of AChE has been elucidated through X-ray crystallo-
graphic studies using Torpedo californica (PDB ID: 1ACJ and 1ACL). These studies re-
vealed that AChE possesses two main binding sites: the catalytic active site (CAS) and the
Drugs Drug Candidates 2023, 2 573
peripheral anionic site (PAS) connected to the gorge within the enzyme’s active site [20–
23]. Amino acids (Trp279, Tyr70, Tyr121, Asp72, and Phe290) form the PAS whereas amino
acids of the esteratic subsite gorge (Ser200, His440, and Glu327), anionic substrate (Trp84,
(Trp279, Tyr70,
Glu199, and Tyr121,
Phe330), andAsp72, and Phe290)
acyl binding pocket form theand
(Phe288 PASPhe299)
whereas aminoform
together acidstheofCAS
the
esteratic
[24–26]. subsite gorge (Ser200, His440, and Glu327), anionic substrate (Trp84, Glu199, and
Phe330),
Theand acyl binding
enzyme pocket (Phe288
acetylcholinesterase is aand Phe299)
serine together
hydrolase form
that the CAS
belongs [24–26].
to the esterase
The enzyme acetylcholinesterase is a serine hydrolase that belongs to the
family within the higher eukaryotes as shown in Scheme 2. This enzyme acts on different esterase
family within the higher eukaryotes as shown in Scheme 2. This enzyme acts on different
types of carboxylic esters. In the cholinergic synapses of the central nervous system (CNS),
types of carboxylic esters. In the cholinergic synapses of the central nervous system (CNS),
AChE hydrolytically cleaves the acetylcholine molecule into choline and acetic acid,
AChE hydrolytically cleaves the acetylcholine molecule into choline and acetic acid, thereby
thereby terminating the impulse transmissions. The interaction between AChE and β-am-
terminating the impulse transmissions. The interaction between AChE and β-amyloid
yloid accelerates the formation of β-amyloid aggregates through the PAS (peripheral ani-
accelerates the formation of β-amyloid aggregates through the PAS (peripheral anionic
onic site). This finding reveals that an ideal AChE inhibitor should be able to interact with
site). This finding reveals that an ideal AChE inhibitor should be able to interact with
both the CAS (catalytic active site) and PAS of the AChE enzyme [27]. Although the exact
both the CAS (catalytic active site) and PAS of the AChE enzyme [27]. Although the
reason for AD is clearly not known, the cholinergic hypothesis proposes a direct correla-
exact reason for AD is clearly not known, the cholinergic hypothesis proposes a direct
tion between the acetylcholine level in the brain, which is hydrolyzed by the enzymes
correlation between the acetylcholine level in the brain, which is hydrolyzed by the enzymes
butyrylcholinesterase and AChE, and the disorder. Acetylcholinesterase, compared to bu-
butyrylcholinesterase and AChE, and the disorder. Acetylcholinesterase, compared to
tyrylcholinesterase, plays an important role in the breakdown of acetylcholine into choline
butyrylcholinesterase, plays an important role in the breakdown of acetylcholine into
and acetic
choline andacid, leading
acetic to the termination
acid, leading of neurotransmission
to the termination signals [28,29]
of neurotransmission signals(Scheme
[28,29]
2).
(Scheme 2).
H3 C
H3 C CH3
H3C OH
N CH3 N
O N H2O CH3
CH3 O
CH3 CH3 OH
O CH3 O Choline O H
H3 C O O NH
NH H3 C NH N
N

..
OH N
..

Estractic site Estractic site

Estractic site
1st transition state Acetyl-AChE
AChE

H3C OH
Acetic acid
O O
H3 C OH
H3C
O N CH H3C OH
3
O CH3 OH NH O NH
H3C N HN
..

Acetylcholine

Estractic site Estractic site

2nd transition state
AChE

Scheme 2.
Scheme 2. Mechanism
Mechanism of
of ACh
ACh hydrolysis
hydrolysis by
by AChE.
AChE.

The
The current approach
approachto toAD
ADmedication
medication focuses
focuses onon designing
designing AChEAChE
andand BuChE
BuChE (bu-
(butylcholinesterase) inhibitors. Cholinesterases (ChEs) have become a
tylcholinesterase) inhibitors. Cholinesterases (ChEs) have become a prominent targetprominent targetin
in
thethe medical
medical field,
field, as most
as most of the
of the U.S.U.S.
Food Food
and and
DrugDrug Administration
Administration (FDA)-certified
(FDA)-certified med-
medications
ications for for
ADAD areare AChE
AChE inhibitors,
inhibitors, includingdonepezil,
including donepezil,galantamine,
galantamine, physostigmine,
physostigmine,
huperzine-A,
huperzine-A, tacrine,
tacrine, and
and rivastigmine
rivastigmine [30,31].
[30,31]. Existing
Existing acetylcholinesterase
acetylcholinesterase inhibitors
inhibitors
typically possess a quaternary or positively ionizable site located within an
typically possess a quaternary or positively ionizable site located within an appropriateappropriate
distance
distancefrom
fromthethe oxygen-containing
oxygen-containingsite site[32,33].
[32,33]. AChE
AChE inhibitors
inhibitors prevent
prevent ACh
ACh hydrolysis
hydrolysis
in
in the neuronal synaptic clefts within the brain, thereby increasing the levelof
the neuronal synaptic clefts within the brain, thereby increasing the level ofACh.
ACh. The
The
prolonged
prolonged use of synthetic cholinesterase inhibitors results in an overall increase in
use of synthetic cholinesterase inhibitors results in an overall increase in the
the
available
available acetylcholine.
acetylcholine. However,
However, excessive
excessive stimulation
stimulation ofof the
the parasympathetic
parasympathetic nervous
nervous
system
systemcancanlead to symptoms
lead to symptoms such assuch
increased hypermotility,
as increased hypersecretion,
hypermotility, bradycardia,
hypersecretion,
miosis, nausea, vomiting, diarrhea, and hypotension, with the potential development
of a cholinergic crisis, which can be life-threatening. Considering these factors, inhibit-
ing acetylcholinesterase is a targeted approach for treating AD and boosting cholinergic
neurotransmission [34–36].
Currently, the drugs available on the market that employ a cholinergic approach for
AD, aiming to increase neurotransmission at cholinergic synapses in the brain and improve
cognition, include tacrine, donepezil, rivastigmine, and galantamine (Scheme 3). However,
 bradycardia, miosis, nausea, vomiting, diarrhea, and hypotension, with the potential de-
velopment of a cholinergic crisis, which can be life-threatening. Considering these factors,
inhibiting acetylcholinesterase is a targeted approach for treating AD and boosting cho-
linergic neurotransmission [34–36].
Drugs Drug Candidates 2023, 2 Currently, the drugs available on the market that employ a cholinergic approach for 574
AD, aiming to increase neurotransmission at cholinergic synapses in the brain and im-
prove cognition, include tacrine, donepezil, rivastigmine, and galantamine (Scheme 3).
However,
these drugsthese
only drugs
provideonly provide symptomatic
symptomatic benefits
benefits and tend andtherapeutic
to lose tend to lose therapeutic
potential over
potential
time [37]. over time [37].

NH2
O

N
N O
N

Tacrine Rivastigmine

O
O
O

O
O
NH
H

Galantamine Donepezil
HO

FDA-approved drugs.
Scheme 3. Structures of FDA-approved drugs.

Therefore,
Therefore, there
there is
is an
an urgent
urgent need
need to to develop
develop aa new
new molecule
molecule that
that can
can address
address the
the
broad
broad spectrum
spectrum of of therapeutic
therapeutic need
need for
for the
the treatment
treatment of
of AD.
AD. The
The focus
focus has
has primarily
primarily been
been
on
on developing
developing inhibitors
inhibitors for
for the
the AChE
AChE enzyme.
enzyme. Researchers
Researchers have
have been
been exploring
exploring other
other
potential target(s) like Butyrylcholinesterase (BChE), amyloid-β (Aβ), β-secretase-1
potential target(s) like Butyrylcholinesterase (BChE), amyloid-β (Aβ), β-secretase-1 (BACE),
and metal
(BACE), ions
and such
metal as such
ions Cu(II),
as Zn(II), or Fe(II),
Cu(II), Zn(II), as wellasaswell
or Fe(II), investigating the antioxidant
as investigating the antiox-
properties and free radical scavenging capacity of compounds in the
idant properties and free radical scavenging capacity of compounds in the last last five years.
five years.
Researchers
Researchers have
have been
been working
working onon establishing
establishing aa well-defined
well-defined pharmacological
pharmacological pro-pro-
file for these compounds by incorporating various substructures within a single
file for these compounds by incorporating various substructures within a single scaffold. scaffold.
Substructures derived from different chemical categories, such as acridine, quinoline, car-
Substructures derived from different chemical categories, such as acridine, quinoline, car-
bamate, huperzine, and other heterocyclic analogs, have been utilized in the development
bamate, huperzine, and other heterocyclic analogs, have been utilized in the development
of these molecules. Structural modifications of prototype molecules have been employed
of these molecules. Structural modifications of prototype molecules have been employed
to optimize their activity through structure–activity relationship (SAR) studies, ultimately
to optimize their activity through structure–activity relationship (SAR) studies, ultimately
leading to the development of novel molecules with expected AChE inhibitory activity
leading to the development of novel molecules with expected AChE inhibitory activity
and other desirable pharmacological properties. The present review outlines the current
and other desirable pharmacological properties. The present review outlines the current
therapeutic strategies in the advancement of these AChE inhibitors in the past two years,
therapeutic strategies in the advancement of these AChE inhibitors in the past two years,
highlighting the progress made in the field.
highlighting the progress made in the field.
2. Design and Synthesis of New AChE Inhibitors during 2021
2. Design and Synthesis of New AChE Inhibitors during 2021
Samaneh Zarei et al., 2021 [38] designed and synthesized the novel series of 1-benzyl-
Samaneh Zarei et al., 2021 methyl)-1-pyridinium
4-((4-oxoquinazolin-3(4H)-yl) [38] designed and synthesized the novel
derivatives seriesin
as shown ofScheme
1-benzyl-4
4-((4-oxoquinazolin-3(4H)-yl)
by methyl)-1-pyridinium
treatment with 3-hydroxy-4-methoxy derivatives
benzoic acid as shown in
and iodomethane in DMF,
Scheme 4 by
which
treatment
yielded with 3,4-dimethoxy
methyl 3-hydroxy-4-methoxy benzoic
benzoate. acid subsequent
Nitration, and iodomethane in DMF,
reduction, which
and cyclisa-
yielded methyl 3,4-dimethoxy benzoate. Nitration, subsequent reduction, and
tion of the above compound yielded 6,7-dimethoxyquina-zolin-4(3H)-one. In the pres-cyclisation
of theofabove
ence K2 COcompound yieldedappropriate
3 and by adding 6,7-dimethoxyquina-zolin-4(3H)-one.
benzyl halide derivatives, In thecompounds
final presence of
were gained by treating the above compound with 3- or 4-(chloromethyl) pyridine in
DMF. Ellman’s method was employed for the evaluation of AChE and BuChE inhibitory
activities. Using Autodock 4, a docking study was carried out to explore the binding
mode of the most active complexes 1 and 2 in the active position of BuChE (PDB: 6QAA)
and AChE (PDB: 1EVE). The compound 1-(3-bromobenzyl)-3-((4-oxoquinazolin-3(4H)-
yl)methyl)pyridin-1-ium bromide (1) was found to be the most active among all synthesized
compounds exhibiting dual inhibition activity for AChE (IC50 = 5.90 ± 0.07 µM) and BuChE
 gained by treating the above compound with 3- or 4-(chloromethyl) pyridine in DMF.
Ellman’s method was employed for the evaluation of AChE and BuChE inhibitory activi-
ties. Using Autodock 4, a docking study was carried out to explore the binding mode of
the most active complexes 1 and 2 in the active position of BuChE (PDB: 6QAA) and AChE
Drugs Drug Candidates 2023, 2 (PDB: 1EVE). The compound 1-(3-bromobenzyl)-3-((4-oxoquinazolin-3(4H)-yl)me- 575
thyl)pyridin-1-ium bromide (1) was found to be the most active among all synthesized
compounds exhibiting dual inhibition activity for AChE (IC50 =5.90 ± 0.07 µM) and BuChE
(IC 50 ==6.76
(IC50 6.76±±0.04
0.04µM).
µM).Further,
Further,the
thecompound
compound1-(4-chlorobenzyl)-3-((6,7-dimethoxy-4-oxo
1-(4-chlorobenzyl)-3-((6,7-dimethoxy-4-oxo
quinazolin-3(4H)-yl)methyl)pyridin-1-ium chloride
quinazolin-3(4H)-yl)methyl)pyridin-1-ium chloride also
also showed
showed potent
potent AChE inhibitory
AChE inhibitory
activity (IC
activity (IC50 = 1.11 ± 0.09 µM) (2). The pharmacokinetic properties of 1 and 2 were studied
50 = 1.11 ± 0.09 µM) (2). The pharmacokinetic properties of 1 and 2 were studied
along with
along with it
it by
by employing
employing the
the Lipinski
Lipinski rules
rules of
of five.
five.

O
O O O
HO
O O O
OH MeI O
HNO3 O O
DMF SnCl2
O
O O NO2 HCl O NH2
3-hydroxy-4-methoxybenzoic acid
methyl 3,4-dimethoxybenzoate
methyl 4,5-dimethoxy-2-nitrobenzoate methyl 2-amino-4,5-
dimethoxybenzoate

N Formadidine acetate
O
Br
O O
N NH
O 3-Chloromethyl
N pyridine
N N+ O N
3-Bromo
6,7-dimethoxyquinazolin-4(3H)-one
1 Br- Benzyl halide O N
O 6,7-dimethoxy-3-((pyridin-3-yl)
methyl)quinazolin-4(3H)-one

4-Chloro
Benzyl halide
O

O
N+ N

Cl
N O
Cl-
2

Scheme 4.
Scheme 4. Synthesis
Synthesis of
of 1-benzyl-4-((4-oxoquinazolin-3(4H)-yl) methyl)-1-pyridinium derivatives.
1-benzyl-4-((4-oxoquinazolin-3(4H)-yl) methyl)-1-pyridinium derivatives.

Mohammad ShahidulShahidul Islam

Islam et al., 2021
2021 [39] designed
designed aa spirooxindole
spirooxindole scaffold
scaffold en-
en-
grafted with two other biologically active molecules such as as indole
indole and
and pyrazole
pyrazole moieties.
moieties.
The authors
authors successfully
successfullysynthesized
synthesized2525derivatives
derivatives and
and screened
screened forfor efficacy
efficacy against
against in-
inhibitory potencytowards
hibitory potency towardsAChE.
AChE.UnderUnderthe thebasic
basiccondition,
condition,new newchalcones
chalconeswere were synthe-
synthe-
sized byby cross
crossaldol
aldolcondensation
condensation ofof
acetyl pyrazole
acetyl pyrazole andand
substituted
substitutedindole-3-carbaldehyde
indole-3-carbalde-
under
hyde under reflux for 72 h. The titled compounds were constructed avia
reflux for 72 h. The titled compounds were constructed via one-pot
a one-potmulticom-
multi-
ponent
component reaction approach
reaction approachin methanol
in methanol under
underreflux conditions
reflux conditions 2−
forfor 24 h.h.A
2−24 A molecular
docking
docking study
studywaswasadopted
adopted to to
explain the the
explain molecular mechanism
molecular mechanism of several lead compounds
of several lead com-
in drug discovery.
pounds The amino
in drug discovery. Theacids
aminoin the enzyme
acids in thesuch as Trp86
enzyme suchand Tyr337and
as Trp86 in the anionic
Tyr337 in
site, Ser203 and His447 in the catalytic site, and Phe295 and Phe297 in the
the anionic site, Ser203 and His447 in the catalytic site, and Phe295 and Phe297 in the acyl acyl binding site
interacted
binding site with compound
interacted and formed
with3compound 3 two
and hydrogen
formed two bonds with Ser125
hydrogen bonds and withGlu202.
Ser125
This
and Glu202. This interaction of the enzyme with compound 3 is important for inhibitory
interaction of the enzyme with compound 3 is important for obtaining good obtaining
activity. Compound
good inhibitory 4 interacted
activity. Compound with4Tyr337 and Phe295
interacted from and
with Tyr337 the anionic
Phe295 and fromacyl
the binding
anionic
sites only but with less hydrophobic interaction than 3. The AChE
and acyl binding sites only but with less hydrophobic interaction than 3. The AChE inhib-inhibitory potency
of compounds
itory and 4 structures
potency of3compounds 3 and 4asstructures
shown inasFigure
shown1 inwas screened
Figure 1 wasby using Ellman’s
screened by using
method,
Ellman’s method, and they exhibited the strongest AChEI with IC50 values ofand
and they exhibited the strongest AChEI with IC 50 values of 24.1 24.1 27.8 µM,
and 27.8
respectively.
µM, respectively.
In continuation of previous research work, Mohammad Shahidul Islam et al., 2021 [40],
via [3+2] the cycloaddition (32CA) reaction starting from the new chalcone, named(E)-3-(5-
chloro-3-methyl-1-phenyl-1H-pyrazol-4-yl)-1-(5-methyl-1-phenyl-1H-pyrazol-4-yl)prop-2-
en-1-one, a novel sequence of spirooxindole analogs tethered to the pyrazole scaffold that
was constructed and synthesized. Acetylcholinesterase inhibitory activity (AChEI) with
IC50 values of 5.7, 7.8, and 8.3 µM was exhibited by compounds 5, 6, and 7, respectively.
The structures of spirooxindole tethered with pyrazole analogs have represented in Figure 2.
Evidently, the addition of the NO2 group into the 5th position and N-methyl glycine (sarco-
sine) took part in the activity, which led to compound 5, the most potent inhibitor with an
 NH NH
H H H

S N N
H H
N N N N
O2 N
Drugs Drug Candidates 2023, 2 576
O O
O O
N
H 3 Cl N
H 4
IC50 value of 5.7 µM. Molecular docking was carried out to determine their interaction with
Figure 1. The structure of compounds 3 & 4: spirooxindole analogues engrafted with Indole and
the active site of hAChE. Via a one-step reaction mechanism with a high polar character,
Drugs Drug Candidates2023,
these 2,Pyrazole
FOR
32CA PEER scaffolds.
REVIEW occurred as a consequence of the super nucleophilic character of
reactions
azomethine yields and the strong electrophilic character of ethylene.
In continuation of previous research work, Mohammad Shahidul Islam et al., 2021
[40], via [3+2] the cycloaddition (32CA) reaction Br
starting from the new chalcone,
Br
named(E)-3-(5-chloro-3-methyl-1-phenyl-1H-pyrazol-4-yl)-1-(5-methyl-1-phenyl-1H-py-
razol-4-yl)prop-2-en-1-one, a novel sequence of spirooxindole analogs tethered to the py-
razole scaffold that was constructed and synthesized. AcetylcholinesteraseNH
inhibitory ac-
NH
H H H
tivity (AChEI) with IC50 values of 5.7, 7.8, and 8.3 µM was exhibited by compounds 5, 6,
and 7, respectively. TheN structures of spirooxindole tethered with N
pyrazole analogs have
S
H H 2 group into the 5th position and
represented
N
in Figure 2. Evidently, the addition of the NO
N N N
O2N N-methyl glycine (sarcosine) took part in the activity, which led to compound 5, the most

potent inhibitorO Owith an IC50 value of 5.7 µM. Molecular Odocking O was carried out to deter-
mine theirN interaction with the active site of hAChE. Via a one-step reaction mechanism
H 3 Cl N
with a high polar character, these 32CA H
reactions occurred 4 as a consequence of the super
Figure 1. The structure of compounds 3 & 4: spirooxindole analogues electrophilic
nucleophilic
Figure character
1. The of azomethine
structure of compoundsyields
3 &and
4: the strong
spirooxindole analogues
engrafted character
engrafted
with of eth-
with
Indole and Indole a
ylene. Pyrazole
Pyrazole scaffolds. scaffolds.

In continuation of previous
N
research work, Mohammad Shahidul Islam et al., 20
[40], via [3+2] the cycloadditionN (32CA) reaction starting from the new chalco
named(E)-3-(5-chloro-3-methyl-1-phenyl-1H-pyrazol-4-yl)-1-(5-methyl-1-phenyl-1H-py
razol-4-yl)prop-2-en-1-one, a novel sequence of spirooxindole analogs tethered to the p
N Cl
O
razole scaffold that was constructed and synthesized. Acetylcholinesterase inhibitory
O 2N
tivity (AChEI) with IC50 values of 5.7, 7.8, and 8.3 µM was exhibited by compounds 5
O Cl
and 7, respectively. The structures of spirooxindole tethered with pyrazole analogs ha
represented in FigureN 2. Evidently, the addition of the NO2 group into the 5th position a
H
N
N-methyl glycine (sarcosine)
5
N took part in the activity, which led to compound 5, the m

potent inhibitor with an IC50 value of 5.7 µM. Molecular docking was carried out to det
mine their interaction with the active site of hAChE. Via a one-step reaction mechani
with a highN polar character, these 32CA reactions
N
occurred as a consequence of the sup
nucleophilic character
N of azomethine yields and
N the strong electrophilic character of e
S ylene.
N Cl N Cl
O O
Br N

O Cl N O Cl

N N
H
N N
N Cl N
6
N O
O 2N 7
Br
O Cl

Figure 2. The structures of compounds N5, 6, & 7: spirooxindole analogs tethered to the pyrazole
Figure 2. The structures of compounds
H 5, 6, & 7: spirooxindole analogs tethered to the pyrazole
scaffold. N
scaffold. N
5

Further, Mohammad Shahidul Islam et al., 2022 [41] designed and synthesized
19 spirooxindole analogs engrafted with pyrazole scaffolds via the [3+2] cycloaddition reac-
tion (32CA) approach throughN the endo/ortho direction. The synthesized
N compounds were
N N
further tested for their antioxidant potentials against DPPH and anti-cholinesterase activity
S
against AChE and BChE. The strucures of compounds 8 and 9 as represented in Figure 3
exhibited IC50 values Cl
of 30 µg/mL for AChE and BChE inhibitoryCl
N O N O activity. With the use of
Br
the Molecular Operating Environment (MOE) software package, the results of invitro and
in silico studies were confirmed,
O Cl the binding mode of synthesized
and O Cl compounds against

the targeted enzymesNH AChE and BChE was determinedNthrough a molecular docking study.
N N
N N
6
7
Br

Figure 2. The structures of compounds 5, 6, & 7: spirooxindole analogs tethered to the pyraz
scaffold.
 were further tested for their antioxidant potentials against DPPH and anti-cholinesterase
activity against AChE and BChE. The strucures of compounds 8 and 9 as represented in
Figure 3 exhibited IC50 values of 30 µg/mL for AChE and BChE inhibitory activity. With
the use of the Molecular Operating Environment (MOE) software package, the results of
invitro and in silico studies were confirmed, and the binding mode of synthesized com-
Drugs Drug Candidates 2023, 2 577
pounds against the targeted enzymes AChE and BChE was determined through a molec-
ular docking study.

Cl F

Cl

N O N O

O O

N N
H H
N N
N N
8 9

Figure 3.
3. The
Thestructure
structureofofcompounds
compounds 8&
8& 9: spirooxindole
9: spirooxindole analogs
analogs engrafted
engrafted withwith pyrazole
pyrazole scaf-
scaffolds.
folds.
Paptawan Suwanhom et al., 2021 [42] designed and synthesized 12quinoxaline deriva-
from o-phenylenediamine
tives Paptawan Suwanhom et al.,derivatives and glyoxal
2021 [42] designed andderivatives
synthesizedin12quinoxaline
ethanol as a solvent
deriv-
by
atives from o-phenylenediamine derivatives and glyoxal derivatives in ethanol as awere
adopting the liquid-assisted grinding (LAG) method. The expected compounds sol-
procured in good the
vent by adopting yields (70–92%) viagrinding
liquid-assisted stannous(LAG)
(II) chloride
method. reduction, and the
The expected 6-amino
compounds
quinoxaline
were procured derivatives were synthesized
in good yields (70–92%) viafrom the corresponding
stannous 2-nitro quinoxalines.
(II) chloride reduction, and the 6-
The structural confirmation waswere
performed 1
by IR, from
H NMR, 13 C NMR, and2-nitro
HRMSquinox-
studies
amino quinoxaline derivatives synthesized the corresponding
and by The
alines. Ellman’s approach
structural using human
confirmation recombinant
was performed byacetylcholinesterase
IR, 1H NMR, 13C NMR, (HuAChE)
and HRMSand
butyrylcholinesterase from equine serum (EqBChE); acetylcholinesterase
studies and by Ellman’s approach using human recombinant acetylcholinesterase inhibitory activity
was evaluated
(HuAChE) andfor the synthesized compounds.
butyrylcholinesterase from equineAll ofserum
the synthesized
(EqBChE);compounds exhibited
acetylcholinesterase
potent inhibitory activity against acetylcholinesterase enzyme
inhibitory activity was evaluated for the synthesized compounds. All of with IC values of 0.077
50 the synthesized
to 50.080 µM. Compared to tacrine
compounds exhibited potent inhibitory 50 (IC = 0.11 µM) and galantamine
activity against acetylcholinesterase50 (IC = 0.59
enzyme µM),
with
compound (12) 2,3-dimethylquinoxalin-6-amine
IC50 values of 0.077 to 50.080 µM. Compared to tacrine (IC = 0.077 µM) had the highest
50 (IC50 = 0.11 µM) and galantamine AChE
inhibitor
(IC50 = 0.59activity in this series.
µM), compound However, some compounds exhibited
(12) 2,3-dimethylquinoxalin-6-amine potent
(IC50 = 0.077 µM)butyryl-
had the
cholinesterase inhibitory activity with IC values ranging from
highest AChE inhibitor activity in this series. However, some compounds exhibitedµM.
50 14.91 to 60.95 A
potent
molecular modeling study was performed using the Discovery Studio
butyrylcholinesterase inhibitory activity with IC50 values ranging from 14.91 to 60.95 µM. 2021 Client and
Visual Molecular Dynamic (VMD) package (AutoDock Tools-1.5.6). The structure of com-
A molecular modeling study was performed using the Discovery Studio 2021 Client and
pounds 10, 11, and 12 as shown in Figure 4, which showed inhibitory activity against AChE,
Visual Molecular Dynamic (VMD) package (AutoDock Tools-1.5.6). The structure of com-
were selected for investigation in detail through molecular docking studies. The dock-
pounds 10, 11, and 12 as shown in Figure 4, which showed inhibitory activity against
ing studies predicted that these compounds bind to the PAS of HuAChE. This molecular
AChE, were selected for investigation in detail through molecular docking studies. The
docking study along with the enzyme kinetic study suggested that compound 12 was a
docking studies predicted that these compounds bind to the PAS of HuAChE. This molec-
mixed-type inhibitor that exhibited uncompetitive AChE inhibition.
ular docking study along with the enzyme kinetic study suggested that compound 12 was
Siju Ellickal Narayanan et al., 2021 [43] aimed at designing and synthesizing various
a mixed-type inhibitor that exhibited uncompetitive AChE inhibition.
brominated derivatives of 7-hydroxy-4-methyl coumarin as a new scaffold (Scheme 5),
which were evaluated for their anti-Alzheimer’s property by in vivo and in vitro models.
The 7-hydroxy 4-methyl coumarin derivatives were synthesized and brominated with a
group of three novel pyrazoles. Based on the docking score of the designed compounds, a
single entity (13) was selected for the treatment of Alzheimer’s disease. Three new pyrazoles
furnished with brominated 7-hydroxy-4-methyl-coumarin derivatives were designed by
Argus lab 4.0.1 version, and the molecular docking studies revealed that compound 13 was
able to bind simultaneously to the amino acid and to the acetylcholine esterase enzyme
active sites. The structure of compound 13 was confirmed by spectral analysis, and by
adopting in vivo and in vitro methods, the anti-Alzheimer’s activity was evaluated; the
obtained results were compared statistically by one-way ANOVA by using GraphPad
Prism. The compound resulted in a significant increase in the acetylcholine esterase level in
acetylcholine esterase inhibition assay. The compound exhibited a promising antioxidant
property based on the DPPH (2,2-diphenyl-1-picryl-hydrazylhydrate) method. Compared
Drugs Drug Candidates 2023, 2 578

to the standard donepezil, the group pretreated with compound 13 exhibited a marked
increase in memory and learning in and elevated plus maze model. Based on its promising
Drugs Drug Candidates2023, 2, FOR PEER REVIEW property and acetylcholine esterase inhibitory activity and MAO inhibitory
antioxidant 8
activity, compound 13 can be marked as a new series among the designed pyrazoles
furnished with brominated 7-hydroxyl-4-methyl coumarin derivatives.

H2N N
H2 N N

N
N
10 11

H2N N CH3

N CH3
Drugs Drug Candidates2023, 2, FOR PEER REVIEW 12 9

Figure 4. The structure of

Figure 4. of compounds
compounds 10,
10,11 &12:
11& Quinoxalinederivatives.
12:Quinoxaline derivatives.

O
Siju Ellickal Narayanan et al., 2021 [43] aimed at designing and synthesizing various
O NH2 O
brominated derivatives of 7-hydroxy-4-methyl
N
coumarin asO a new scaffold (Scheme 5),
H
which were evaluated for their anti-Alzheimer’s property by in vivo and in vitro models.
H C
The 37-hydroxy 4-methyl coumarin derivatives were O synthesized
N NH and brominated with a
Br
group of threeOnovel pyrazoles. Based on the docking score of the designed compounds,
a singleBr entity (13) was selected for the treatment of Alzheimer’s disease. Three new pyra-
zoles furnished O with brominated 7-hydroxy-4-methyl-coumarin derivatives were de-
signed by Argus lab 4.0.1 version, and the molecular
2-(3-bromo-4-methyl-2-oxo-2H-chromen-7-yloxy)acetohydrazide (E)-ethyl 2-(2-(2-bromophenyl)hydrazono)-3-oxobutanoate
docking studies revealed that com-
pound 13 was able to bind simultaneously to the amino acid and to the acetylcholine es-
terase enzyme active sites. The structure of compound 13 was confirmed by spectral anal-
ysis, and by adopting in vivo and in vitro Omethods, the anti-Alzheimer’s activity was eval-
uated; the obtained results were compared statistically by one-way ANOVA by using
O N
GraphPad Prism. The compound resulted inNa significant increase in the acetylcholine es-
CH3
terase level in acetylcholine esterase inhibition assay. The compound exhibited a promis-
H3C
ing antioxidant property based on the DPPH O (2,2-diphenyl-1-picryl-hydrazylhydrate)
method. Compared to the standard donepezil, theN groupNH pretreated with compound 13
O 13 Br
Br
exhibited a marked increase in memory and learning in and elevated plus maze model.
Based on its promising antioxidant O property and acetylcholine esterase inhibitory activity
and MAO inhibitory activity, compound 13 can be marked as a new series among the
designed pyrazoles furnished with brominated 7-hydroxyl-4-methyl coumarin coumarin deriva-
with pyrazole.
pyrazole.
Scheme 5. Synthesis of brominated derivatives of 7-hydroxy-4-methyl coumarin with
tives.
Lukas Gorecki et al., 2021 [44] reported a series of 30 novel tacrine derivatives derivatives out- out-
lined for the evaluation of structure–activity relationships and
lined for the evaluation of structure–activity relationships and studied the receptors using studied the receptors using
electrophysiology
electrophysiology with with HEK293
HEK293 cells cells expressing
expressing the the defined
defined typestypes of of NMDARs.
NMDARs. Com- Com-
pounds
pounds 14 14 and
and 15, 15, which
whichwere wereselected,
selected,effectively
effectivelyinhibitedinhibitedboth bothGluN1/GluN2A
GluN1/GluN2A and and
GluN1/GluN2B
GluN1/GluN2B receptors. receptors. The The GluN1/GluN2B
GluN1/GluN2Breceptors receptors were were more
more effectively
effectively inhibited
inhibited
by
by compounds
compounds 16 16 and
and 17. The structure
17. The structure of of potent
potent AChE AChE inhibitors
inhibitors are are shown
shown in in Figure
Figure 5.
5.
At a concentration of 100 µM, GluN1/Glu2B were inhibited
At a concentration of 100 µM, GluN1/Glu2B were inhibited at −60 mV, expressed as the at − 60 mV, expressed as the
IC
IC50 value, and there was relative inhibition of GluN1/Glu2A at +40 mV based on a QSAR
50 value, and there was relative inhibition of GluN1/Glu2A at +40 mV based on a QSAR
study.
study. The The statistically
statistically significant
significant model model revealed
revealed by by thethe QSAR
QSAR study study can can bebe employed
employed
for
for a ligand-based virtual screening and to determine the potent molecule for inhibition
a ligand-based virtual screening and to determine the potent molecule for inhibition
of
of GluN1/Glu2A and/or GluN1/Glu2B subtypes. The tested compounds did not not
GluN1/Glu2A and/or GluN1/Glu2B subtypes. The tested compounds did cause
cause hy-
hyperlocomotion in an open field and also did not impair
perlocomotion in an open field and also did not impair the prepulse inhibition of the star- the prepulse inhibition of the
tle response compared to MK-801 when in vivo experiments were carried out in rats but
did show minimal induction of psychotomimetic side effects. Overall, it is confirmed that
tacrine derivatives are encouraging as they are centrally available subtype-specific inhib-
itors of NMDARs without any complications.
 GluN1/GluN2B receptors. The GluN1/GluN2B receptors were more effectively inhibited
by compounds 16 and 17. The structure of potent AChE inhibitors are shown in Figure 5.
At a concentration of 100 µM, GluN1/Glu2B were inhibited at −60 mV, expressed as the
IC50 value, and there was relative inhibition of GluN1/Glu2A at +40 mV based on a QSAR
study. The statistically significant model revealed by the QSAR study can be employed
Drugs Drug Candidates 2023, 2 for a ligand-based virtual screening and to determine the potent molecule for inhibition 579

of GluN1/Glu2A and/or GluN1/Glu2B subtypes. The tested compounds did not cause hy-
perlocomotion in an open field and also did not impair the prepulse inhibition of the star-
startle response
tle response compared
compared to MK-801
to MK-801 whenwhen in vivo
in vivo experiments
experiments werewere carried
carried out out in rats
in rats but
but
did did
showshow minimal
minimal induction
induction of psychotomimetic
of psychotomimetic sideside effects.
effects. Overall,
Overall, it is confirmed
it is confirmed that
that tacrine
tacrine derivatives
derivatives are encouraging
are encouraging as are
as they they are centrally
centrally available
available subtype-specific
subtype-specific inhib-
inhibitors of NMDARs
itors of NMDARs without
without any complications.
any complications.

NH2 NH2

Br Br

N N
14 15

NH2 NH2

Cl

Drugs Drug Candidates2023, 2, FOR PEER REVIEW N N 10

16 17

Figure 5. The
Figure 5. The structure
structure of
of compounds 16 &
compounds 16 17: Tacrine
& 17: Tacrinederivatives.
derivatives.
Muhammad Mansha et al., 2021 [45] synthesized a series of 13 new fluoroquinolone
Muhammad
derivatives Mansha their
and evaluated et al.,AChE
2021 [45]
andsynthesized a series
BChE inhibitory of 13 new
activities. fluoroquinolone
Through FT-IR, 1H,
derivatives and evaluated their AChE and BChE inhibitory activities. Through
and 13C NMR, as well as elemental analysis, structural characterization was performed.
13 FT-IR, 1 H,
and C NMR,
Compound 18,as7-chloro-1-cyclopropyl-6-fluoro-N-(2-(2-
well as elemental analysis, structural characterization was performed.
fluorobenzamido)ethyl)-4-oxo-
Compound 18, 7-chloro-1-cyclopropyl-6-fluoro-N-(2-(2- fluorobenzamido)ethyl)-4-oxo-1,4-
1,4-dihydroquinoline-3- carboxamide shown in Figure 6 showed the highest inhibitory
dihydroquinoline-3-carboxamide
activity shown in
(2.20 ± 0.10 mM) as it contained theFigure
fluoro6 substituent
showed the athighest inhibitory
the ortho activity
position. The
(2.20 ± 0.10 mM) as it contained the fluoro substituent at the ortho position. The inhibitory
inhibitory activity against BChE was decreased slightly due to the positioning of the fluoro
activity against BChE was decreased slightly due to the positioning of the fluoro substituent
substituent to the para position. The activity was further decreased when the fluoro sub-
to the para position. The activity was further decreased when the fluoro substituent was
stituent was placed in the meta position. The SAR study was used to explore those com-
placed in the meta position. The SAR study was used to explore those compounds that
pounds that contained electronegative functional groups such as F, Cl, OMe, N, and O at
contained electronegative functional groups such as F, Cl, OMe, N, and O at the ortho
the ortho position of the phenyl group, which exhibited greater inhibitory activities as
position of the phenyl group, which exhibited greater inhibitory activities as compared
compared to their meta and/or para substituents. Molecular docking studies of the syn-
to their meta and/or para substituents. Molecular docking studies of the synthesized
thesized compounds revealed that some compounds docked into the active site of AChE,
compounds revealed that some compounds docked into the active site of AChE, and
and few compounds with BChE showed p–p interactions along with conventional H
few compounds with BChE showed p–p interactions along with conventional H bonding
bonding with the active residues of AChE through their electronegative functions and
with the active residues of AChE through their electronegative functions and phenyl ring,
phenyl ring, respectively.
respectively.

O O
H
O N F
N
H
F
18 N Cl

Figure 6.6.The
Thestructure
structure of compound
of compound 18: 7-chloro-1-cyclopropyl-6-fluoro-N-(2-(2-
18: 7-chloro-1-cyclopropyl-6-fluoro-N-(2-(2- fluoroben-
fluorobenzamido)
zamido)ethyl)- 4-oxo-1,4-dihydroquinoline-3- carboxamide.
ethyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide.

Aljohanie etetal.,
Ghadah Aljohanie al.,2021
2021[46],
[46],who
who used
used thethe Claisen–Schmidt
Claisen–Schmidt condensation
condensation re-
reaction,
action, synthesized
synthesized 12 novel
12 novel chalcones
chalcones by treating
by treating 2-alkyloxy-naphthaldehydes
2-alkyloxy-naphthaldehydes and
and man-
nich bases of 4-hydroxyacetophenone with a catalytic amount of SOCl2 in ethanol to ob-
tain the chalcones with good yield. Structural confirmation was performed using spectro-
scopic analysis using FTIR, 1D and 2D NMR, and HRMS spectroscopy. Molecular docking
was carried out to study the ligand interactions with the target to highlight its affinity. In
Drugs Drug Candidates 2023, 2 580

mannich bases of 4-hydroxyacetophenone with a catalytic amount of SOCl2 in ethanol

to obtain the chalcones with good yield. Structural confirmation was performed using
spectroscopic analysis using FTIR, 1D and 2D NMR, and HRMS spectroscopy. Molecular
docking was carried out to study the ligand interactions with the target to highlight its
affinity. In addition to the docking functions, the computational biology study highlighted
the structure–activity relationship (SAR) and pharmacokinetic properties (absorption,
distribution, metabolism, excretion, and toxicity or ADMET) of the potential ligands. To
screen their affinity towards the AChE enzyme (PDB 1EVE), comparative docking analysis
was carried out. The synthesized chalcones exhibited lower binding energy (−13.06 to
−10.43 kcal/mol) against AChE compared to donepezil (−10.52 kcal/mol). The study on
SAR explored the introduction of diethyl amine in ring A of the structure of the chalcone
skeleton,
Drugs Drug Candidates2023, 2, FOR PEER REVIEW and the propargyl moiety of ring B is known to be a potential drug against AD.11
The complex nature of chalcone 19 shown in Figure 7 was proven to be advantageous,
indicating an effective and promising drug against AChE.

HO O

N O
19
Figure7.7.The
Figure Thestructure
structureof
ofcompound 19:Chalcone
compound19: Chalconederivative.
derivative.

3. Design and Synthesis of New AChE Inhibitors during 2022

3. Design and Synthesis of New AChE Inhibitors during 2022
In a study conducted by Zaizafoon Nabeel et al., 2022 [47], novel benzo[f]coumarin
In a study conducted by Zaizafoon Nabeel et al., 2022 [47], novel benzo[f]coumarin
derivatives with a pyrimidine unit were synthesized successfully as represented in Scheme 6.
derivatives with a pyrimidine unit were synthesized successfully as represented in
In the alkaline medium, the benzo[f]coumarin chalcone was prepared by condensing 3-
Scheme 6. In the alkaline medium, the benzo[f]coumarin chalcone was prepared by con-
acetyl-5,6-benzocoumarin and 4-hydroxybenzaldehyde Via Claisen–Schmidt condensation.
densing 3-acetyl-5,6-benzocoumarin and 4-hydroxybenzaldehyde Via Claisen–Schmidt
By the cycloaddition of chalcone with urea, thiourea, and guanidine HCl and in the pres-
condensation. By the cycloaddition of chalcone with urea, thiourea, and guanidine HCl
ence of glacial acetic acid, various pyrimidines were synthesized. Through spectral and
and in the presence of glacial acetic acid, various pyrimidines were synthesized. Through
elemental analyses, the structures of the newly synthesized compounds were character-
spectral
ized. To and elemental
assess analyses,
the potential the structures
of the synthesized of the newly synthesized
compounds as AChE compounds
inhibitors, an were
in
characterized. To assess the potential of the synthesized compounds
silico study was carried out using molecular docking simulation. Discovery Studio Vi- as AChE inhibitors,
an in silico
sualizer wasstudy was carried
employed out using
to determine themolecular
foremost docking
binding simulation.
mode with the Discovery
receptor Studio
and
Visualizer was employed to determine the foremost binding mode with
obtain 3D interaction poses. The docking simulations of the compounds exhibited greater the receptor and
obtain 3D interaction poses. The docking simulations of the compounds
electrostatic interactions (van der Waals, π-π stacking, and H bonds) with lower docking exhibited greater
electrostatic
energies interactions
(−17.4, 13.6, and (van
−11.3 der Waals, π-π
kcal/mol) stacking,
compared to and H bonds)score
the docking withoflower docking
the standard
energies (−17.4, 13.6, and −11.3 kcal/mol) compared to the docking
donepezil (−10.6 kcal/mol). Compound 3 demonstrated four H-bond interactions with score of the standard
donepezil
key (−10.6
amino acid kcal/mol).
residues, Compound
including 3 demonstrated
Ser235, Asn230, Pro232, four H-bond
and His398.interactions with key
Other electrostatic
amino acid were
interactions residues, including
observed Ser235,
between Asn230, 20
derivatives Pro232,
and 21and andHis398.
residuesOther electrostatic
Cys402, Trp524,
and Ala234, indicating similar binding interactions with the active pockets of theTrp524,
interactions were observed between derivatives 20 and 21 and residues Cys402, AChE
and Ala234,
enzyme. indicatingthe
Furthermore, similar binding
compounds interactions
were screened with the AChE
for their active inhibitory
pockets ofactivity
the AChEus-
enzyme.
ing Furthermore,
Ellman’s method, with thedonepezil
compounds were
as the screened
standard. The forEllman’s
their AChE
assayinhibitory activity
results disclosed
using
that Ellman’s method,
derivatives 21 and 22 with
haddonepezil as the standard.
AChE inhibitory activity and The revealed
Ellman’s higher
assay results dis-
inhibition
closed that derivatives 21 and 22 had AChE inhibitory activity and revealed higher inhi-
bition percentages at the concentrations of 10–4 and 10–10 M while a lower inhibition per-
centage was obtained at 10–12.
Drugs Drug Candidates 2023, 2 581

Drugs Drug Candidates2023, 2, FOR PEER REVIEW 12

Drugs Drug Candidates2023, 2, FOR PEER REVIEW 12
percentages at the concentrations of 10–4 and 10–10 M while a lower inhibition percentage
was obtained at 10–12 .
CHO CH
CHO CH33
O CHO
O CHO
OH
OH O
O
CH
CH33
C H O O
C22H55O O
2-hydroxynaphthalene-1- ethyl 3-oxobutanoate O O
2-hydroxynaphthalene-1-
carbaldehyde ethyl 3-oxobutanoate O O
carbaldehyde 2-acetyl-3H-benzo[f]chromen-3-one
2-acetyl-3H-benzo[f]chromen-3-one
OH
OH
X 4-hydroxybenzaldehyde
X 4-hydroxybenzaldehyde

O
HN N O
HN N
1. Urea
1.
2. Urea
Thourea
2.
3. Thourea
Guanidine
3. Guanidine

O O OH
HO O O O O OH
HO O O 2-((E)-3-(4-hydroxyphenyl)acryloyl)-3H-benzo[f]chromen-3-one
2-((E)-3-(4-hydroxyphenyl)acryloyl)-3H-benzo[f]chromen-3-one
X = O (20), S (21), NH (22)
X = O (20), S (21), NH22(22)

Scheme 6. Synthesis of
Scheme of novel benzo[f]coumarin
benzo[f]coumarin derivatives bearing substituted pyrimidine.
Scheme 6. Synthesis
Synthesis of novel
novel benzo[f]coumarin derivatives bearing substituted pyrimidine.

AA series
A series of of novel
novel sunifiram–carbamate
sunifiram–carbamate and and sunifiram–anthranilamide
sunifiram–anthranilamide hybrids
sunifiram–anthranilamide hybrids were
hybrids were
designed and
designed
designed and synthesized
synthesized by by Khalid
Khalid A. A.Agha
Agha et
Agha et al.,
et al., 2022
al., 2022 [48]
2022 [48]
[48] andand evaluated
and evaluated
evaluated for for cholinergic
forcholinergic
cholinergic
activity. The authors concluded that introducing
activity. The authors concluded that introducing carbamate to the skeleton
activity. carbamate to the skeleton
skeleton of of synthesized
of synthesized
synthesized
targets resulted
targets
targets resultedin ingood
in goodAChE
good AChEinhibitory
AChE inhibitoryactivity.
inhibitory activity.Novel
activity. Novelsunifiram
Novel sunifiram
sunifiram analogs
analogs
analogs thatthat
that are
areare
ableable
able to
to
to modulate
modulate
modulate N-methyl-D-aspartate
N-methyl-D-aspartate
N-methyl-D-aspartate ReceptorsReceptors
(NMDARs)
Receptors (NMDARs)
(NMDARs) and areand and are
equipped equipped with
with the structural
are equipped with the the
structuralthat
features
structural features that
enablethat
features them enable
to inhibit
enable themacetylcholinesterase
them to inhibit
to inhibit acetylcholinesterase
acetylcholinesterase
enzyme were enzyme were con-
constructed.
enzyme were con-A
structed.
carbamate A carbamate
moiety as moiety
well as as
2- or well as 2-
4-aminophenylor 4-aminophenyl
and 3-pyridyl
structed. A carbamate moiety as well as 2- or 4-aminophenyl and 3-pyridyl moieties aided and 3-pyridyl
moieties moieties
aided the aided
AChEI
the AChEI
the AChEI
activity activity
of activity
the of the
targets.
of the
The targets.
most The
targets. The most
potent mostAChEpotent AChE inhibitory
inhibitory
potent AChE inhibitory
activity with activity
an IC
activity with an IC
50 value
with an ICof50
50
value
18 ± of
0.2 18
nM ± 0.2
was nM was
exhibited exhibited
by the by the
target target
compound: compound:
the
value of 18 ± 0.2 nM was exhibited by the target compound: the sunifiram–carbamate hy- the sunifiram–carbamate
sunifiram–carbamate hybrid hy-
23.
brid 23.
Such
brid 23. Suchof
ability
Such ability of compound
compound
ability of compound23 (Figure 23 (Figure
23 (Figure
8) along8) along
8) along
with withwith
its its favorable
favorable
favorable
its log
log plog pp value
value value (2.36),
(2.36),
(2.36),its
its ability
ability to to
induceinduce
ACh ACh
release release
from from
A549 A549
cells, cells,
and its and
in its
vivo
its ability to induce ACh release from A549 cells, and its in vivo ability to lower AChE in vivo
ability ability
to lower to lower
AChE AChE
activity
activity
in the rat
activity inbrain
in the rat
the rat brainitmakes
makes
brain makes
worthy itit worthy
worthy
of further of further
of further investigation
investigation as aa promising
as a promising
investigation as promising
nootropic nootropic
agent.
nootropic
agent.
Compared Compared
to to
sunifiram sunifiram
( − 4.5 kcal/mol)(−4.5 kcal/mol)
and the and the
anthranilamide
agent. Compared to sunifiram (−4.5 kcal/mol) and the anthranilamide hybrid (−4.5 anthranilamide
hybrid ( − 4.5 hybrid
kcal/mol), (−4.5 it
kcal/mol),
exhibited a it exhibited
molecular a molecular
docking score −
docking
1.7 score
kcal/mol −1.7
whenkcal/mol
kcal/mol), it exhibited a molecular docking score −1.7 kcal/mol when docked to the gly- docked whento docked
the to the
glycine-binding gly-
cine-binding
pocket pocket
of the pocket
cine-binding NMDAof of the NMDA
receptor.
the NMDA receptor.receptor.

O
O

N
N

N
N O
O
(23)
(23) O
O

SS NN
H
O
O
2-methylthioethyl Side
Side chain
chain H
2-methylthioethyl
is optimum
is optimum for
for inhibiting
inhibiting AChE
AChE

Introduction of
Introduction of Carbmate
Carbmate Succed
Succed in
in
inhibition of AChE
inhibition of AChE
Figure 8. The structure of compound 23: sunifiram carbamate hybrid.
hybrid.
Figure 8. The structure of compound 23: sunifiram carbamate hybrid.

Makar Makarian
Makar Makarian et
et al.,
al., 2022
2022 [49]
[49] planned
planned and
and synthesized
synthesized aa series
series of
of 12
12 donepezil-
donepezil-
based analogs with moderate yield by adopting an environmentally friendly route,
based analogs with moderate yield by adopting an environmentally friendly route, i.e.,i.e., by
by
employing microwave irradiation, and the structural purity of the
employing microwave irradiation, and the structural purity of the synthesized synthesized
Drugs Drug Candidates 2023, 2 582

Drugs Drug Candidates2023, 2, FOR PEER REVIEW 13

Makar Makarian et al., 2022 [49] planned and synthesized a series of 12 donepezil-
based analogs with moderate yield by adopting an environmentally friendly route, i.e., by
employing microwave irradiation, and the structural purity of the synthesized compounds
compounds
was confirmed wasbyconfirmed by spectral
spectral analysis. analysis.
With donepezilWithasdonepezil
a standard, asthe
a standard, the com-
synthesized syn-
thesizedwere
pounds compounds were tested
tested against electricagainst electric
eel AChE eel AChE
enzyme using enzyme
Ellman’susing Ellman’s spec-
spectrophotometric
trophotometric
method for their method for their
inhibitory inhibitory
activity. The SARactivity.
studyThe SAR study 24
of compound of compound
as mentioned 24 asin
mentioned in Figure 9 showed that through aromatic π- π stacking
Figure 9 showed that through aromatic π- π stacking interactions, an indanone moiety interactions, an inda-
none1)
(Site moiety
bound (Site 1) bound
to the to theanionic
peripheral peripheral anionic
site (PAS) ofsite (PAS)AChE,
enzyme of enzyme and AChE, and the
the piperidine
ring (Site 2)ring
piperidine exhibited
(Site 2)an interaction
exhibited with the anionic
an interaction with thepart of the
anionic catalytic
part active site,
of the catalytic i.e.,
active
with the with
site, i.e., amino the acid tyrosine
amino (Y337). The
acid tyrosine benzyl
(Y337). Themoiety
benzylof donepezil
moiety (Site 3) was
of donepezil (Sitein3)close
was
proximity to tryptophan
in close proximity (W86), and
to tryptophan two amino
(W86), and two acids, H447
amino and H447
acids, S203, were both part
and S203, wereofboth
the
catalytic triad. It was clear from the SAR study that alteration at site
part of the catalytic triad. It was clear from the SAR study that alteration at site 3 could3 could lead to the
development of potent AChE
lead to the development inhibition,
of potent that is, thethat
AChE inhibition, replacement of the benzyl
is, the replacement moiety
of the benzyl at
site 3 with
moiety the 3pyridine
at site with thering couldring
pyridine be acould
potentbeinhibitor
a potentofinhibitor
electric eel AChE. eel AChE.
of electric

N
Site 1 O Site 3
O

Site 2

O
(24)

Figure 9.
Figure 9. The
The structure of compound
structure of compound 24.
24. Donepezil-based
Donepezil-based analog.
analog.

Matosevicetetal.,al.,
Ana Matosevic 20222022 [50][50] designed
designed and and synthesized
synthesized 18 biscarbamate
18 biscarbamate compoundscom-
pounds
with with substituents
various various substituents in the carbamoyl
in the carbamoyl and hydroxyaminoethyl
and hydroxyaminoethyl chain, and chain,
theirand
in-
hibitory potential and inhibition selectivity were determined toward
their inhibitory potential and inhibition selectivity were determined toward both cholin-both cholinesterases.
Confirmation of the structure
esterases. Confirmation of theofstructure
the compounds was performed
of the compounds was through
performed NMR and HRMS
through NMR
spectra.
and HRMS Forspectra.
all of the
Forsynthesized compounds,
all of the synthesized the potential
compounds, theofpotential
biscarbamates to reduce
of biscarbamates
the activitythe
to reduce of activity
human of BChE
human and BChE
AChEand wasAChE
examined. The synthesized
was examined. compounds
The synthesized in-
com-
hibited
pounds BChE, which
inhibited proves
BChE, which these compounds
proves are fast orare
these compounds very
fastfast
or BChE inhibitors
very fast with
BChE inhib-
ki constants 6 M−1 min6 −1 .−1The molecule
itors with ki in the range
constants in of
the(0.0144–38.0)10
range of (0.0144–38.0)10 M min−1. The molecule with piperidine
with piper- in
the carbamoyl and hydroxyaminoethyl chain was confirmed to
idine in the carbamoyl and hydroxyaminoethyl chain was confirmed to be compound 25, be compound 25, which
exhibited the fastest
which exhibited inhibition,
the fastest being an
inhibition, almost
being 10 times
an almost 10faster
timesinhibitor than bambuterol.
faster inhibitor than bam-
But the compounds
buterol. 25 and 2625asand
But the compounds depicted in Figurein
26 as depicted 10Figure
that inhibited BChE 1288
10 that inhibited and 1087
BChE 1288
times faster
and 1087 than
times AChE
faster were
than AChE the were
most the
selective. Ultimately,
most selective. the study
Ultimately, therevealed that on
study revealed
the
thatbenzene ring, biscarbamates
on the benzene with a meta
ring, biscarbamates withdisposition of carbamate
a meta disposition groups could
of carbamate be
groups
acould
favorable core base for the design of new AD drugs. Compared
be a favorable core base for the design of new AD drugs. Compared to the standardto the standard drug
rivastigmine, compound
drug rivastigmine, compound26 actively inhibited
26 actively BChEBChE
inhibited and is supposed
and to beto
is supposed non-toxic and
be non-toxic
be able to pass the BBB and have the capacity to chelate biometals. Finally,
and be able to pass the BBB and have the capacity to chelate biometals. Finally, a study a study singled
out compounds
singled 25 and 26
out compounds 25asand the26most promising
as the for the treatment
most promising of AD. of AD.
for the treatment
Drugs Drug
Drugs Drug Candidates
Candidates2023, 2, FOR PEER
2 FOR
2023, 2,
Candidates2023, PEER REVIEW
REVIEW 14
583
14

OH
OH OH
OH
O O N H
H
O O N O
O O
O N
N

HCl
HCl HCl
N
N HCl
N
N

(25) O
O O N
(25) (26)
(26) O N

O
O O
O
Figure 10.
Figure
Figure 10. The
10. The structure
The structure of
structure of compounds
of compounds 25
compounds 25 &
25 & 26.
& 26. Biscarbamates
26. Biscarbamates derivatives.
Biscarbamates derivatives.
derivatives.

Radhika Kachhadiya
Radhika Kachhadiya et et al.,
al., 2022
2022 [51]
[51] designed
designed and and synthesized
synthesized novel
novel quinazolinone
quinazolinone
derivatives, and the synthesized compounds were screened
derivatives, and the synthesized compounds were screened for their purity andfor their purity and structural
structural
characterization by
characterization by spectral
byspectral studies
spectralstudies
studies toto
to actact
act as potent
as potent
as potentanti-Alzheimer’s
anti-Alzheimer’s
anti-Alzheimer’s agents. In silico
agents.
agents. In silico dock-
In dock-
silico
ing studies
ing studies
docking werewere
studies
were performed employing
performed
performed Autodock
employing
employing 4.2 for
Autodock
Autodock 4.2 for the
4.2the synthesized
forsynthesized compounds
the synthesized compounds
compounds 27–
27–
29 to
to observe
27–29
29 observe
to observethethe
the binding interactions
binding
binding interactions
interactions compared
comparedto
compared totothe
the standard
thestandard donepezil.
standarddonepezil.
donepezil. AsAs compared
As compared
compared
to the
to the standard
standarddonepezil,
standard donepezil,the
donepezil, thebinding
the bindingpocket
binding pocket
pocket of
ofof the
thethe
AChEAChE
AChE compounds
compounds
compounds 27–29
27–29 exhibited
exhibited
27–29 exhibitedno
no
no less
lessless
thanthan one
oneone
than hydrogen
hydrogen
hydrogen bond
bond
bond interaction
interaction
interaction with
with Phe
withPhe A:295.
PheA:295. Docking
A:295.Docking studies
Dockingstudies showed
studies showed
showed thatthat
compounds 30–32
compounds 30–32 exhibited
exhibited two
two hydrogen
hydrogen bonding
bonding interactions
interactions with Tyr A:124
with Tyr A:124 andand Phe
Phe
A:295. The
A:295. The compounds
compounds were were subjected
subjected to to ADMET
ADMET studies,
studies, andand by
by using
using Swiss
Swiss ADME
ADME and and
pk
pk CSM
pk CSM software,
CSM software, physicochemical
software, properties
physicochemical properties
physicochemical were
properties were also
also predicted.
were also Compounds 27–32
predicted. Compounds
predicted. as
27–32 as
as
shown in Figure
Figure 11
11 exhibited
exhibited better
better pharmacokinetic
pharmacokinetic profiles and
shown in Figure 11 exhibited better pharmacokinetic profiles and are known to exhibit are known
known to
to exhibit
exhibit
cholinesterase inhibitory
cholinesterase inhibitory
inhibitory activity
activity against
activity against Alzheimer’s
against Alzheimer’s disease.
Alzheimer’s The synthesized
disease. The
disease. synthesized
synthesized novel
novel
novel
quinazolinone derivatives are known to exhibit good BBB
quinazolinone derivatives are known to exhibit good BBB and CNS permeation,
quinazolinone and CNS permeation, a
permeation, a low low
volume
volume of
volume of distribution,
of distribution, and
distribution, and no
and no negative
no negative effect
negative effect on
effect onrenal
on renalclearance.
renal clearance.
clearance.

R22
R
O
O
Compo. R11
R R22
R
Compo.
H
H
N
N
N
N 27
27 2,4-dichloro
2,4-dichloro 4-Methyl
4-Methyl
28
28 4-Chloro
4-Chloro 4-Methoxy
4-Methoxy
N NH 29
29 4-Methyl
4-Methyl 4-Methyl
4-Methyl
N NH
30
30 2-Methyl
2-Methyl 4-Methoxy
4-Methoxy
31
31 4-Chloro
4-Chloro 2-Chloro
2-Chloro
R11
R 32
32 4-Chloro
4-Chloro 4-Methyl
4-Methyl

Figure 11.
Figure 11. The
The structure
structure of
of compounds
compounds 27
27 to
to 32:
32: Quinazolinone
Quinazolinone derivatives.
derivatives.

Xinnan
XinnanLi
Xinnan Lietet
Li etal.,
al.,2022
al., [52]
2022
2022 designed
[52]
[52] designed
designedandandsynthesized
and by fusing
synthesized
synthesized natural
by fusing
by fusing (±)-7,8-dihydroxy-
natural
natural (±)-7,8-dihy-
(±)-7,8-dihy-
3-methyl-isochroman-4-one
droxy-3-methyl-isochroman-4-one pharmacophore
pharmacophorewith donepazil
with to
donepazil
droxy-3-methyl-isochroman-4-one pharmacophore with donepazil to achievea series achievea
to series
achievea of novel
series of
of
isochroman-4-one
novel isochroman-4-onederivatives as
derivativesgiven
as in Scheme
given in 7,
Schemeand their
7, and anticholinesterase
their
novel isochroman-4-one derivatives as given in Scheme 7, and their anticholinesterase po- potential
anticholinesterase po-
against
tential Alzheimer’s
tential against
against diseasedisease
Alzheimer’s
Alzheimer’s was evaluated.
disease Compound
was evaluated.
was evaluated. 33 [(Z)-3-acetyl-1-benzyl-4-((6,7-
Compound
Compound 33 [(Z)-3-acetyl-1-benzyl-
33 [(Z)-3-acetyl-1-benzyl-
dimethoxy-4-oxoisochroman-3-ylidene)methyl)
4-((6,7-dimethoxy-4-oxoisochroman-3-ylidene)methyl) pyridin-1-ium
4-((6,7-dimethoxy-4-oxoisochroman-3-ylidene)methyl) pyridin-1-ium bromide]
pyridin-1-ium had average
bromide]
bromide] an-
had aver-
had aver-
tioxidant
age activityactivity
age antioxidant
antioxidant and showed
activity potent acetylcholinesterase
and showed
and showed (AChE) inhibitory
potent acetylcholinesterase
potent acetylcholinesterase activity asactiv-
(AChE) inhibitory
(AChE) inhibitory well.
activ-
Molecular
ity as well.modeling
Molecular and kinetic
modeling investigations
and kinetic demonstrated
investigations that compound
demonstrated
ity as well. Molecular modeling and kinetic investigations demonstrated that compound that33 exhibited
compound
a33dual
33 binding
exhibited
exhibited inhibitory
aa dual
dual binding
bindingproperty and property
inhibitory
inhibitory bound
property to and
both CAS and
and bound
bound PAS
both of
to both
to CAS
CAStheand
enzyme
and PAS AChE.
PAS of the
of the
In silico screening
enzyme AChE.
enzyme AChE. In revealed
In silico that
silico screening compound
screening revealed 33
revealed that could across
that compound
compound 33 the blood–brain
33 could
could across barrier
across the with
the blood–
blood–
high
brainpenetration.
brain barrier withInhigh
barrier with addition,
high low cytotoxicity
penetration.
penetration. In addition,
In andlow
addition, moderate
low anti-Aβ
cytotoxicity
cytotoxicity and
andaggregation efficacy
moderate anti-Aβ
moderate anti-Aβ
were exhibited
aggregation
aggregation by thewere
efficacy
efficacy compound.
were exhibited
exhibited Finally,thethe
by the
by study revealed
compound.
compound. Finally,
Finally, that
thecompound
the 33 from
study revealed
study revealed a
that
that
 Drugs Drug Candidates2023, 2, FOR PEER REVIEW 15
Drugs Drug
Drugs Drug Candidates
Candidates2023,
2023, 2,
2 FOR PEER REVIEW 15
584

compound 33 from a natural product exhibited a promising lead toward the development
compound
natural 33 from
product a natural
exhibited product exhibited
a promising a promising
lead toward lead towardofthe
the development development
a potent AChE
of a potent AChE inhibitory molecule in the treatment of AD.
of a potentmolecule
inhibitory AChE inhibitory moleculeofinAD.
in the treatment the treatment of AD.
Natural product analog
Natural product analog
O
O
HO
HO

O
O O O
HO O O
HO
6,7-dihydroxy-3-methyl-1H-isochromen-4(3H)-
6,7-dihydroxy-3-methyl-1H-isochromen-4(3H)- O
one O
one
Br-
Fusion Br -
Fusion O N+

O O N+
O Low Toxicity
Low Toxicity Anti-AChE
AChE Inhibitor N (33) Anti-AChE
Antioxidant
AChE Inhibitor N (33) Antioxidant
O
O
O
O

O
O Donepezil
Donepezil

Scheme 7. Synthesis of [(Z)-3-acetyl-1-benzyl-4-((6,7-dimethoxy-4-oxoisochroman-3-ylidene)me-

Scheme 7.7.Synthesis
Synthesis of [(Z)-3-acetyl-1-benzyl-4-((6,7-dimethoxy-4-oxoisochroman-3-ylidene)me-
thyl) pyridin-1-iumofbromide].
Scheme [(Z)-3-acetyl-1-benzyl-4-((6,7-dimethoxy-4-oxoisochroman-3-ylidene)methyl)
thyl) pyridin-1-ium bromide].
pyridin-1-ium bromide].
Namy George et al., 2022 [53] designed and synthesized coumarin-linked 1,3,4-oxadi-
Namy George
Namy George etetal.,al.,
20222022[53][53]
designed
designedand synthesized
and coumarin-linked
synthesized coumarin-linked1,3,4-oxadi-
1,3,4-
azole hybrid derivatives(as multi-target directed ligands (MTDLs)). The synthesized com-
azole hybridhybrid
oxadiazole derivatives(as
derivatives(asmulti-target directeddirected
multi-target ligands (MTDLs)).
ligands The synthesized
(MTDLs)). The com-
synthe-
pounds were investigated for their in vitro AChE, BuChE inhibitory activity, antioxidant
pounds
sized were investigated
compounds for their in for
were investigated vitro AChE,
their BuChE
in vitro inhibitory
AChE, BuChEactivity,
inhibitoryantioxidant
activity,
activity, and cyclooxygenase (COX) activity using standard spectrophotometric methods.
activity, and cyclooxygenase (COX) activity using standard spectrophotometric
antioxidant activity, and cyclooxygenase (COX) activity using standard spectrophotomet- methods.
Their chemical structures were characterized using analytical data. With the help of online
Their
ric chemicalTheir
methods. structures
chemical werestructures
characterizedwereusing analytical using
characterized data. With the help
analytical of online
data. With
cheminformatics software, the binding mode of the synthesized compounds with AChE
cheminformatics
the help of onlinesoftware,
cheminformaticsthe binding mode of
software, thethe synthesized
binding mode of compounds with AChE
the synthesized com-
and BuChE in addition to the pharmacokinetic profile was predicted by molecular dock-
and BuChE
pounds withinAChE
additionandto the pharmacokinetic
BuChE in addition to the profile was predicted
pharmacokinetic by molecular
profile dock-
was predicted
ing studies. Compounds 34 and 35 as mentioned in Figure 12 (were found to be the most
ingmolecular
by studies. Compounds
docking studies. 34 andCompounds
35 as mentioned 34 andin Figure 12 (were found
35 as mentioned to be 12
in Figure the(were
most
potent AChE inhibitors (IC50 values = 29.56 and 28.68 µM) among the tested compounds
potentto
found AChE
be theinhibitors
most potent (IC50AChE
values = 29.56 and
inhibitors (IC5028.68
valuesµM) among
= 29.56 andthe tested
28.68 µM)compounds
among the
for anticholinesterase activity. Compared to the standard galantamine (SI = 1.132), com-
for anticholinesterase
tested compounds for activity. Comparedactivity.
anticholinesterase to the standard
Compared galantamine
to the standard(SI = galantamine
1.132), com-
pounds 34 and 35 also showed a higher selectivity index (SI) of 1.652 and 1.552. From
pounds
(SI 34 and
= 1.132), 35 also showed
compounds 34 and a35higher selectivity
also showed index selectivity
a higher (SI) of 1.652 and (SI)
index 1.552.of From
1.652
molecular docking studies, it is clear that compounds 34 and 35 bound well to AChE
and 1.552. docking
molecular From molecular
studies, docking
it is clear studies, it is clear that
that compounds 34 compounds
and 35 bound 34 and
well 35to bound
AChE
(binding
well to AChE energy scores of 9.7scores
(binding and 10.19.7 Kcal/Mol) and were identified toidentified
be the two most
(binding energy scores energy
of 9.7 and 10.1ofKcal/Mol) and 10.1 andKcal/Mol) and were
were identified to be the two to be
most
potent
the twoAChE AChE inhibitors
most potent AChE through
inhibitors in vitro
through assay. The in vitro antioxidant and anti-inflam-
potent inhibitors through in vitro assay.inThe vitroinassay. The in vitro and
vitro antioxidant antioxidant and
anti-inflam-
matory activity studies
anti-inflammatory activity revealed
studies that that thethat
revealed synthesized hybridhybrid
the synthesized molecules also exhibited
molecules also ex-
matory activity studies revealed the synthesized hybrid molecules also exhibited
good-to-excellent
hibited results. Based on the in vitro study and molecular docking study, cou-
good-to-excellent results. Based on the in vitro study and molecular docking study,study,
good-to-excellent results. Based on the in vitro study and molecular docking cou-
marin oxadiazole
coumarin oxadiazole hybrids
hybrids act as MTDLs and are aa promising
promisingsourcesourceofofanti-AD
anti-ADdrugs;
drugs;
marin oxadiazole hybrids actact
as as MTDLs
MTDLs andand arearea promising source of anti-AD drugs;
coumarin
coumarin oxadiazole hybrids can lead to the development of highly potent therapeutics
coumarin oxadiazole
oxadiazolehybrids
hybridscan canlead
leadtotothethedevelopment
development of of
highly
highlypotent
potent therapeutics
therapeutics for
for
the the treatment
treatment of AD. of AD.
for the treatment of AD.
O OH O
O OH O
HO HO
HO O O
O HO
O

HO O O HO O O
HO O O HO O O

N N
N N 34 N N 35
N 34 N 35
Figure 12. The structure of compounds&34 &Coumarin-linked
35: Coumarin-linked 1,3,4-oxadiazole hybrid deriva-
Figure
Figure 12.12. The
Thestructure
structureofofcompounds
compounds3434 35:
& 35: 1,3,4-oxadiazole
Coumarin-linked hybrid
1,3,4-oxadiazole derivatives.
hybrid deriva-
tives.
tives.
Drugs Drug
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FORREVIEW
PEER REVIEW 16 16
Drugs Drug Candidates 2023, 2 585

Evangelia-Eirini N. Vlachou
Evangelia-Eirini et al., et
N. Vlachou 2022
al.,[54]
2022synthesized bis-fused
[54] synthesized pyridopyranocou-
bis-fused pyridopyranocou-
marinsEvangelia-Eirini
by using goldN. Vlachou et
nanoparticles al., 2022
supported[54] synthesized
on TiO bis-fused
employing
marins by using gold nanoparticles supported on TiO2 employing microwave
2 pyridopyranocoumarins
microwave irradiation
irradiation
by using
through gold
the nanoparticles
triple bond supported
activation of on TiO employing
propargylaminopropargyloxy
2 microwave
through the triple bond activation of propargylaminopropargyloxy coumarins irradiation
coumarins or through
propar-
or propar-
the triple bond activation
gylamino-fused
gylamino-fused of propargylaminopropargyloxy
pyranocoumarins.
pyranocoumarins.Via the Viapropargylation coumarins
the propargylation or propargylamino-
of 6-amino-7-hydroxycouma-
of 6-amino-7-hydroxycouma-
fused
rins pyranocoumarins.
orrins Via the propargylation
6-amino-4-hydroxycoumarin,
or 6-amino-4-hydroxycoumarin, the new new of
thepropargyl 6-amino-7-hydroxycoumarins
aminopropargyloxycoumarin,
propargyl aminopropargyloxycoumarin, oran6- an
amino-4-hydroxycoumarin,
intermediate, was synthesized.
intermediate, the new
was synthesized. propargyl
High Highanti-lipid aminopropargyloxycoumarin,
peroxidation
anti-lipid activities
peroxidation an intermedi-
were presented
activities were presentedby by
ate, was synthesized.
compound 36 whereasHigh anti-lipid
compound 37 peroxidation
is a new and activities
promising were presented
class with by compound
antioxidant and
compound 36 whereas compound 37 is a new and promising class with antioxidant and
36 whereasactivities.
anti-AChE compound 37 is a new
Compound 36andcan36promising
becan
used class with antioxidant and anti-AChE
anti-AChE activities. Compound be as
useda lead
as amultifunctional molecule
lead multifunctional as it as it
molecule
activities. antilipid
combines Compound 36 can be with
peroxidation used interesting
as a lead multifunctional
anti-AChE molecule
activity. as it combines
Compound 37 is a37 is a
combines antilipid peroxidation with interesting anti-AChE activity. Compound
antilipid peroxidation
new anti-AChE with
lead. Compoundsinteresting anti-AChE
36 and3637and fully activity. Compound 37 is a new anti-
new anti-AChE lead. Compounds 37obeyed Lipinski’s
fully obeyed rule ofrule
Lipinski’s fiveofand
fivestruc-
and struc-
AChE
ture lead.
ofture
these Compoundsdepicted
compounds 36 and 37 in fully
Figure obeyed
13. Lipinski’s rule of five and structure of
of these compounds depicted in Figure 13.
these compounds depicted in Figure 13.
O O

O O

O O O OO O

H2 N O O
H2 N
36 36 N 37 37
N

Figure 13.
13. The
FigureFigure structure
13.
The of
of compounds
The structure
structure 36
36 &
& 37:
of compounds
compounds 36Bis-fused
37: pyridopyranocoumarins
& 37: Bis-fused
Bis-fused derivatives.
pyridopyranocoumarins
pyridopyranocoumarins derivatives.
derivatives.

Ibadulla
Ibadulla Mahmudov
Mahmudov
Ibadulla Mahmudov et
et al., 2022
al.,et al., [55],
2022 2022by
[55], by adapting
adapting
[55], new
new methods,
by adapting methods,
new methods,synthesized
synthesized six
synthesizedsixN-
N-six N-
allyl and N-benzyl
allyl allyl and N-benzyl aniline derivatives.
anilineaniline
derivatives. For the
For theFor
derivatives. alkylation
alkylation reaction,
reaction,
the alkylation allyl bromide
allyl bromide
reaction, and ben-
and benzyl
allyl bromide and ben-
zyl chloride
chloridezyl were were
taken
chloride taken
were as starting
as starting
taken material,
asmaterial,
starting andand halide
halide
material, was
and was the
the ion
halide was ion scavenger.
scavenger.
the Triethyla-
Triethylamine
ion scavenger. Triethyla-
mine
was madewas
minemade useunder
use made
was of ofuse
under the
theunder
of reflux
refluxthe condition
condition
reflux ofofN,N-dimethyl
condition N,N-dimethyl acetamide
acetamide
of N,N-dimethyl (DMA).
(DMA).
acetamide The
The The
(DMA).
synthesized
synthesized compound
compound N-benzyl
N-benzyl and
and N-allyl
N-allyl aniline
aniline derivatives
derivatives were
were
synthesized compound N-benzyl and N-allyl aniline derivatives were screened for their screened
screened for
for their
their
inhibitory
inhibitory activity
activity
inhibitory against
against
activity AChE
AChE
against and
and
AChE carbonic
carbonic
and anhydrases
carbonic anhydrases (hCAs).
anhydrases (hCAs).The The
(hCAs). compound
Thecompound
compound N-ben-N-
N-ben-
benzyl
zyl and and N-Allyl
N-Allyl anilineaniline derivatives
derivatives exhibited
exhibited good good inhibitory
inhibitory activity
activity
zyl and N-Allyl aniline derivatives exhibited good inhibitory activity against AChE withagainst against
AChE AChE
with
with
IC IC
50 values
IC50 values
of 489.98
50 values
of 489.98
of 489.98
nM and nM
nMKiand Ki
Ki values
values
and of
of 424.69
of 424.69
values ± 98.56
424.69 ± 98.56Among
± nM.
98.56 nM. Among
nM. Among the synthesized
synthesized
the synthesized
the com- com-
compounds,
pounds, N-benzyl-4-chloroaniline
N-benzyl-4-chloroaniline (38, (38, Figure
Figure 14) 14) showed
showed a a good
good
pounds, N-benzyl-4-chloroaniline (38, Figure 14) showed a good inhibitory effect inhibitory
inhibitory effect
effect on on
on
AChE.
AChE. In
In order
order to
to support
support the
the experimental
experimental in
in vitro studies for
AChE. In order to support the experimental in vitro studies for both hCAs and AChE both hCAs and AChE
AChE
inhibitors,
inhibitors, in silico
in
inhibitors,silico molecular
molecular
in silico docking
docking
molecular computations
computations
docking computationswere performed
were performedwith thewith Autodock
Autodock
the Autodock
Vina program, and there was a good correlation with the
Vina program, and there was a good correlation with the experimental experimental results.
results.
results.

HN HN Cl Cl

38 38
Figure 14. The
FigureFigure
14. The structure
structure
14. of compound
of compound
The structure 38:
of compoundN-benzyl-4-chloroaniline.
38: N-benzyl-4-chloroaniline.
38: N-benzyl-4-chloroaniline.

Abduljelil Ajala
Abduljelil Ajala et
et al.,
al., 2022
2022 [56] made use
useofofstructure-based drug
drugdesign techniques to
Abduljelil Ajala et al.,[56]
2022made[56] made structure-based
use of structure-based design techniques
drug design techniques
create 15
to create hydrazone
15 hydrazone derivatives used
derivatives to evaluate
used to their
evaluate acetylcholinesterase inhibitor
their acetylcholinesterase potency
inhibitor
to create 15 hydrazone derivatives used to evaluate their acetylcholinesterase inhibitor
to treat Alzheimer’s
potency to treat disease. The
Alzheimer’s protein
disease. The target (code
protein ID 4EY7)
target (code was
ID selected
4EY7) onselected
was the basisonof
potency to treat Alzheimer’s disease. The protein target (code ID 4EY7) was selected on
an extensive
the basis literature survey. Based on the obtained protein code after analysis, a protein
the of an extensive
basis literature
of an extensive survey.
literature BasedBased
survey. on theonobtained protein
the obtained code after
protein code anal-
after anal-
target was drawn to interact with the 15 hydrazone derivatives containing piperazine
ysis, aysis,
protein target was drawn to interact with the 15 hydrazone derivatives
a protein target was drawn to interact with the 15 hydrazone derivatives containing containing
39 as shown in Figure 15 (compounds with a higher binding energy). Critical guidance
piperazine 39 as shown
piperazine 39 of in Figure
astheir
shown 15 (compounds
in Figure 15 (compounds with awith
higher binding energy).
a higher Critical
offered on the basis stronger interactions, higher binding scores, binding
enhanced energy).
drug-likeCritical
guidance offered
guidance on theonbasis
offered the of their
basis of stronger
their interactions,
stronger higherhigher
interactions, binding scores,scores,
binding en- en-
properties, and drug kinetic parameters allowed the synthesis of novel potent hydrazone
hanced drug-like
hanced properties,
drug-like and drug kinetic
properties, parameters allowed the synthesis of novel
derivatives, and a protein target wasand drug kinetic
designed parameters
to interact allowed
with these the synthesis
compounds of novel
of interest.
 Drugs
Drugs Drug Drug Candidates2023,
Candidates2023, 2, FOR
2, FOR PEER PEER REVIEW
REVIEW 17 17

Drugs Drug Candidates 2023, 2 586

potent potent hydrazone

hydrazone derivatives,
derivatives, and a protein
and a protein target
target was was designed
designed to interact
to interact with these
with these
The compounds
compounds
molecular of
of interest.interest.
docking The The molecular
molecular
studies docking
revealed docking studies
studies revealed
the important revealed the
theresidues
active site important
important active
active site
involved in site
the residues
binding
residues involved
interactions
involved in the
in theofbinding binding
the most interactions
potent compound
interactions of the
39 with
of the most most potent
the compound
potent compound
4EY7 receptor, 39
scoring
39 with thewith the
27.23 4EY7
4EY7 kcal/mol.receptor, scoring
The studies
receptor, scoring 27.23 kcal/mol.
revealedThe
27.23 kcal/mol. The
thatstudies studies
the potent revealed
compound
revealed that
that the can the potent
be compound
potent compound
used to create
can can
be usedbe
promisingtoused topromising
create promising
pharmacotherapeutic
create pharmacotherapeutic
compounds in treating
pharmacotherapeutic compounds
AD.
compounds in treating
in treating AD. AD.

O Attachement
Attachement of para position
of para position on the on the
O Phenyl ring plays arole
crucial
Phenyl ring plays a crucial in role in
increasing
increasing activity activity
N+ N+
-
-
O O
39 39
N N
N N HN HN

N N
O O

Figure Figure
Figure 15.
15. The 15. The structure
The structure
structure of of compound
of compound
compound 39: Hydrazone
39: Hydrazone
39: Hydrazone derivative
derivative
derivative containing
containing
containing piperazine.
piperazine.
piperazine.

Bhushanarao
Bhushanarao
Bhushanarao Dogga et Dogga
et al., et al.,
al., 2022
2022 2022
[57]
[57] [57] designed,
designed,
designed, synthesized,
synthesized,
synthesized, and evaluated
andevaluated
and evaluated a series
a series aofseries of
of
11 11 11
analogs analogs
analogsof of of tacrine-2-amide
tacrine-2-amide
tacrine-2-amide forfor for anti-AD.
anti-AD.
anti-AD. Out Out Out
of of
11 11 of 11 analogs,
analogs,
analogs, compounds
compounds
compounds 40,40, 40,
41,41,
andand41,
42 and 42
42 (Figure
(Figure
(Figure 16)16) 16)
showed
showed showed
good
good good activity at concentrations
activityatatconcentrations
activity concentrationsofof23.66 of
23.66nM,23.66
nM,20 nM,
20 nM, 20
and 24.33 nM, nM,
nM, and nM, and 24.33
respectively,
respectively,
respectively, whereas,
whereas, whereas,
the
the other
othertheeight
other
eight eight derivatives
derivatives
derivatives showedshowed
showed moderate
moderate moderate
activityactivity
activity at
at less at less than
less than
than
100 nM. In silico study revealed that these 11 analogs can strongly bind to both the CASthe CAS
100 nM. 100
In nM.
silico In silico
study study
revealed revealed
that that
these 11 these 11
analogs analogs
can can
strongly strongly
bind to bind
both to
the both
CAS
and
and PAS
PASand
of PAS
of an
an of an enzyme.
enzyme.
enzyme. Docking
Docking Docking
studies studies
studies clearly clearly revealed
clearly revealed
revealed that that compound
that compound
compound 40 showed
40 40 showed
showed good good
good
affinity affinity toward AChE with a binding energy of 24.03
affinity toward AChE with a binding energy of 24.03 kcal/mol and had a glide score of
toward AChE with a binding energy of 24.03 kcal/mol kcal/mol
and had and
a had
glide a glide
score ofscore of
10.38 kcal/mol.
10.38 Also,
kcal/mol. compound
Also, compound 40 exhibited
40 exhibited
π-π stacking
π-π with
stacking Trp86
10.38 kcal/mol. Also, compound 40 exhibited π-π stacking with Trp86 and showed an in- an in-
with and
Trp86 showed
and showedan
interaction
teraction withTyr341.
teraction
with Tyr341. Similarly,
with Tyr341.
Similarly, compound
Similarly,
compound compound
41 hada41
41had aglide
glide score
had ascore
glideof −11.22
ofscore
−11.22 kcal/mol
of kcal/mol
−11.22 anda and a
kcal/mol
and
abinding
binding energy
binding of −
energy 30.88
of kcal/mol,
−30.88 which
kcal/mol, was
which contributed
was by
contributed pi–pi
energy of −30.88 kcal/mol, which was contributed by pi–pi stacking interaction bystacking
pi–pi interaction
stacking interaction
with Trp86
with and a
Trp86 hydrogen
and a bond
hydrogen with
bondthe amino
with the group
amino
with Trp86 and a hydrogen bond with the amino group of tacrine with His447. of tacrine
group ofwith His447.
tacrine with His447.

NH2 NH2
40 40
H H
N N N N

N N O O

O O
N N

O O

NH2 NH2
N N
41 41
N N
N 42 42
N
NH2 NH2 N N
N N

O O O O

N N

Figure Figure
Figure 16.
16. The 16. The structure
The structure
structure of of compound
of compound
compound 40,
40, 41 & 40,
41 & 42: 41
42: & 42: tacrine-2-amide
tacrine-2-amide
tacrine-2-amide derivatives.
derivatives.
derivatives.

4. Conclusions
In conclusion, the most effective strategy for the enhancement of ACh would be an
effective method to compensate for the deficiency of acetylcholine, focusing on enhanc-
Drugs Drug Candidates 2023, 2 587

ing cholinergic neurotransmission. For the effective treatment of Alzheimer’s disease,

cholinesterase inhibitors are the only pharmacological agents that proved to be applica-
ble. The first and foremost enzyme responsible for the breakdown of ACh in the nervous
system is AChE; the activity of AChEIs is characterized by the inhibition of AChE. This
inhibition of AChE prolongs the action of the insufficient neurotransmitter in the brain.
For the symptomatic treatment of AD, AChEIs were the first drugs licensed, and three
compounds associated with this therapeutic class, i.e., donepezil (Aricept), rivastigmine
(Exelon), and galantamine (Razadyne), are currently in clinical use in many countries
worldwide. However, vomiting, nausea, diarrhea, bradycardia, abnormal dreams, and
fatigue are a few of the severe side effects of these FDA-approved drugs that are associ-
ated with AD. As a consequence of AD’s complexes, there is an urgent need to develop
effective therapeutic agents with minimal side effects to effectively combat AD. Due to its
hepatotoxicity, tacrine (the fourth AChE inhibitor) is no longer in use. For the therapy of
adult-onset dementia disorders, the combination with other classes of drugs may be an
opportunity for a renewed interest in AChEIs. Other approaches along with the AChEIs in
combination with other classes of drugs are cholinergic precursors, N-methyl-d-aspartate
(NMDA) receptor antagonists, and antioxidant agents.
In the past few decades, many AChEIs have been tested in clinical trials for the
treatment of AD. In limiting the progression of cognitive impairment, a few of them
have been successful in terms of showing a beneficial effect on daily living activities in
AD patients. However, none of the experimental AChEI compounds reported superior
efficacy and lesser complexity than those currently available for the treatment of AD, thus
discouraging advanced research in the field.
In this review, we discussed the design, synthesis, biological activity, and molecular
docking studies on acetylcholinesterase inhibitors. To act as a potent anti-Alzheimer’s agent,
the criteria for a novel molecule should be clearly understood from an extensive literature
survey, and a molecule should be designed followed by its synthesis. The synthesized
compounds should be confirmed by subjecting them to spectral studies, such as IR, MASS,
and NMR spectroscopic methods, and then characterized. Compared to standard drugs, the
potency of the synthesized compound in inhibiting the enzyme acetylcholinesterase should
be cleared. SAR studies and in silico docking studies are to be carried out to determine the
binding mode and its interactions with the synthesized compounds.
In this study, 30 novel tacrine derivatives, 12 novel chalcones, 12quinoxaline deriva-
tives, a new series of 13fluoroquinolone derivatives, 6N-allyl and N-benzyl aniline deriva-
tives, a series of tacrine-2-amide derivatives, and novel benzo[f]coumarin derivatives
bearing the pyrimidine unit were successfully outlined and synthesized. Along with these
various brominated derivatives of 7-hydroxy coumarin as a new scaffold, 25synthesized
hits based on a spirooxindole scaffold engrafted with two other pharmacophores, including
indole and pyrazole moieties, were designed, synthesized, and evaluated for their anti-AD
activity. Through the triple bond activation of propargylaminopropargyloxycoumarins
or propargylamino-fused pyranocoumarins, bis-fused pyridopyranocoumarins were syn-
thesized in excellent yield by gold nanoparticles supported on TiO2 under microwave
irradiation. Coumarin-connected 1,3,4-oxadiazole hybrid derivatives were also designed
and synthesized (as multi-target directed ligands (MTDLs). All of the synthesized molecules
exhibited good results as novel molecules. These discoveries led to the development of
potential molecules as anti-Alzheimer agents with the involvement of available drugs and
active compounds such as acetylcholinesterase inhibitors. It can be concluded that many
more novel potential molecules with good inhibitory potency and lesser side effects are yet
to be discovered in the upcoming years. A computational perspective can be valuable in the
design of compounds with better pharmacological profiles combined with the structural
information available for inhibitors with diverse chemical complexity.

Author Contributions: S.L.M.: conceptualization, methodology. writing—review draft. B.P.N.:

Validation, conceptualization, and supervision. M.S.K.: visualization, investigation, writing—review,
editing, and supervision. All authors have read and agreed to the published version of the manuscript.
Drugs Drug Candidates 2023, 2 588

Funding: This research received no external funding.

Conflicts of Interest: The authors declare no conflict of interest.

Abbreviations

AD Alzheimer’s disease
Ach Acetylcholine
AChE Acetylcholinesterase
AChEIs Acetylcholinesterase Inhibitors
Ab b-amyloid peptide
BChE Butylcholinesterase
BACE β-secretase-1
CNS Central Nervous System
ChAT Choline O-acetyltransferase
CAS Catalytic active site
COX cyclooxygenase
PAS peripheral anionic site
SAR Structure–activity relationships
TcAChE Torpedo californica AChE
MAO Monoamine Oxidase
BBB Blood–Brain Barrier
NMDARs N-methyl-D-aspartate Receptor
DMF Dimethyl formamide
DPPH α-diphenyl-β-picrylhydrazyl

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