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1662 Current Medicinal Chemistry, 2013, 20, 1662-1672

Rationally Designed Multi-Targeted Agents Against Neurodegenerative

Diseases

W. J. Geldenhuys and C. J. Van der Schyf*

Department of Pharmaceutical Sciences, College of Pharmacy, Northeast Ohio Medical University, Rootstown, OH
44272, USA

Abstract: Neurodegenerative diseases are complex disorders with several pathoetiological pathways leading to cell death.
Rationally designed multi-targeted agents, or “multi-targeted designed drugs” (MTDD) show significant promise in pre-
clinical studies as neuroprotective and disease-modifying agents. In this review, we highlight the use of chemical scaffolds
that lend themselves exquisitely to the development of MTDDs in neurodegeneration. Notably, synthetic polycyclic cage
compounds have served as scaffolds for novel voltage-gated calcium channel blockers, NMDA receptor antagonists, and
sigma-receptor ligands – attractive targets in neurodegeneration. In an entirely different approach, compounds containing
the thiazolidinedione moiety (referred to as glitazones) alter mitochondrial function through the mitochondrial protein mi-
toNEET, an attractive new drug target for the treatment of neurodegenerative diseases. The design strategy for yet another
agent, ladostigil, employed the amalgamation of active chemical moieties of the AChE inhibitor rivastigmine, and the
monoamine oxidase-B (MAO-B) inhibitor rasagiline, leading to a single compound that targets both enzymes simultane-
ously. Natural products have also served as design templates for several MTDD design studies. In particular, the stilbene
scaffold has become popular in particular due to the neuroprotective effects of the non-flavonoid natural product resvera-
trol. Recently, stilbene scaffold-based compounds were developed to reduce – through chelation with metal ions that in-
teract with beta-amyloid – both metal-induced beta-amyloid protein aggregation, and ROS generated from this aggregate.
Other subtle modifications of the stilbene motif led to the creation of reversible, non-competitive MAO inhibitors. Finally,
compounds derived from the xanthine scaffold afford neuroprotection in Parkinson’s disease through mechanisms that in-
clude dual adenosine A2A receptor antagonism and MAO-B inhibition.
Keywords: Multifunctional agents, multi-targeted designed drugs, neurodegenerative diseases, drug discovery.

1. INTRODUCTION would be akin to the discovery of antibiotics in the treatment

As our society ages, we are confronted with an increase
in age-related disorders including neurodegenerative diseases
such as Alzheimer’s and Parkinson’s disease, Lewy body
disease, post-stroke degeneration, and frontotemporal de-
mentias among others. Patients who suffer from these neu-
rodegenerative diseases, have increased morbidity and mor-
tality, that in turn also affect society through significantly
increased health care costs [1], negatively impacting the
economies of entire countries. Currently, treatment options
for patients diagnosed with neurodegenerative diseases are
limited. Disease modifying therapeutics have yet to emerge
from current research endeavors. Essentially, those drugs
currently approved only afford symptomatic relief from the
disease. For example, Parkinson’s disease treatment consists
of approaches aimed at correcting the loss of fine motor con-
trol, but affords little or no mitigation of the progression of
the disease. These medications do not alter the rate or extent
of neuronal cell loss [2, 3]. As far as drug discovery in neu-
rodegeneration goes, the development of disease-modifying
agents for the treatment of neurodegenerative disorders
*Address correspondence to this author at the Department of Pharmaceuti-
cal Sciences, College of Pharmacy, Northeast Ohio Medical University,
Rootstown, OH 44272, USA; Tel: +1-330-325-6467; Fax: +1-330-325-
5936; E-mail: cvanders@neomed.edu
of bacterial infections.
What constitutes the major reasons for the fact that no
disease-modifying agents have been brought to the clinic to
date? This simple question may relate to the methods by
which drug discovery endeavors have been practiced in the
last century. Pharmaceutical companies were traditionally
engaged in developing compounds that display a single ac-
tivity profile at very high potency – a single “magic bullet”
which would interact with a singular drug target [4, 5]. In
contrast - with neurodegeneration - this concept has not met
with expected success such as accomplished, for example, by
treating hypertension with calcium channel antagonists [5].
A contrasting but alternative strategy is proposed that pur-
ports to target multiple drug intervention points in complex
diseases such as neurodegeneration to fundamentally alter
disease progression – a “shotgun” effect [5, 6].
Over the past few years, several reviews have indicated
that this approach is leading to a paradigm shift in drug de-
velopment for neurodegenerative diseases [1, 4, 5, 7-9]. Ad-
ditionally, several studies have now been published in which
active drug design techniques were used to construct multi-
ple-ligand or multifunctional drugs. Morphy and Rankovic
tallied the number of reports describing this active discovery
of designed multiple-ligand drugs (See Fig. 1). Between
1994 and 2004 over 300 reports were published in the

1875-533X/13 $58.00+.00 © 2013 Bentham Science Publishers

Rationally Designed Multi-Targeted Agents
Traditional Drug Design
Methods
Drug 1
Drug Target 1 Drug Target 1
Treatment
Fig. (1). The development of therapeutic agents has traditionally been focused on one drug target, but the complexity of neurodegenerative
disease has necessitated a new approach whereby one chemical agent is designed to act via several disease pathways.
medicinal chemistry literature regarding the design of com-
pounds targeted to more than one rational drug target. The
diseases covered encompassed many conditions, most nota-
bly cancer and neurodegeneration. Both cancer and neurode-
generation have several common themes, of which multiple
compensatory mechanisms underlie the basis of multifunc-
tional drug design [10].
In neurodegenerative disease, much of the complexity be-
tween linked signaling pathways is still poorly understood.
In the case of neurodegenerative disorders, the targeting of a
single disease pathway may therefore not be sufficient to
modulate the cell death process due to compensatory mecha-
nisms that reside outside the influenced of the drug. Several
studies have indicated that the complexity of neurodegenera-
tion requires a multifaceted approach [6, 10]. Additionally,
by using a single chemical moiety, the probability of serious
side effects is lessened, as compared to the classical poly-
pharmaceutical approach that requires the use of multiple
drug molecules in the treatment of patients [1].
In this review, we will consider select examples of de-
signed multiple ligand drugs that have been evaluated
against several drug targets currently thought to play a role
in the cell death cascades of neurons. Several challenges
exist in the development of designed multiple ligand drugs.
These include: protein targets that are key players in the cell
death cascade; finding a balance in the affinity for each re-
spective target (i.e. the magnitude of affinity or activity
needs to be pharmacokinetically relevant); for CNS drugs,
the ability to traverse the blood-brain barrier (BBB). The
latter issue is of particular importance, because for CNS
drugs, a large proportion of novel compounds fail to reach
the clinic due to poor CNS penetration [11].
2. CALCIUM CHANNEL MODULATORS
Calcium plays an important role in normal neuronal
physiology [12-14]. However, under pathological conditions,
an over-accumulation of calcium inside neurons may trigger
cell death signaling cascades. Two major conduits for cal-
cium entry into neuronal cells include voltage gated calcium
channels (VGCC) and the N-methyl-D-aspartate receptor/ion
channel (NMDAR). Both these ion channels have been asso-
ciated with neuronal cell death and have been considered
Current Medicinal Chemistry, 2013, Vol. 20, No. 13 1663
MTDD
Drug 2
Drug Target 2 Drug Target 3
Treatment
drug targets to prevent or attenuate neuronal cell death [13,
15]. Additionally, several studies have suggested that a com-
bination of VGCC and NMDAR blockers may be beneficial
as a disease-modifying strategy as compared to blocking
either of these ion channels in isolation [16, 17].
Memantine is an adamantanamine with a polycyclic cage
structure (Fig. 2), approved for the treatment of moderate to
severe Alzheimer’s disease (Trade name: Namenda®) [3,
18]. Memantine is a derivative of the anti-parkinsonian and
antiviral agent amantadine [19], and has been extensively
studied over the past few decades in the treatment of neu-
rodegeneration, both for Alzheimer’s and Parkinson’s dis-
ease [20, 21]. From electrophysiological studies, it was
shown that memantine has favorable NMDA modulation
activity due to its fast on-off kinetic profile at the receptor.
There, in the presence of memantine, the ion channel of the
NMDAR is normalized in ion conductance as opposed to
MK-801 (dizocilpine) or phencyclidine (PCP), both of which
block the channel completely – an effect that leads to hallu-
cinations and cell death [20]. From in vivo studies such as
from stroke animal models, it has been suggested that the
combination of memantine (an NMDAR antagonist) and
nimodipine (a VGCC blocker) is more beneficial than the
respective individual therapies [22]. The putative positive
effects of this combined therapy led to the development of
NGP1-01, a polycyclic cage amine, as a dual NMDAR and
VGCC blocker [23-25].
Prior to the introduction of amantadine in 1966 as a pro-
phylactic agent against Asian influenza, polycyclic cage
structures have not been explored for pharmacological prop-
erties [19]. The pentacycloundecane (PCU) cage structure
became prominent in the 1980s after the group of Van der
Schyf et al., demonstrated that their PCU-derived lead com-
pound, NGP1-01, blocks calcium influx into cells, specifi-
cally the L-type VGCC [25] (Fig. 2). Initially, the structural
data of NGP1-01 was interpreted to contain an aza-
bridgehead in the cage (Fig. 2), but X-ray crystallography
soon suggested that the cage was indeed oxa-bridged.
The PCU is a versatile scaffold that can be used effec-
tively for the modification of a compound’s lipophilicity
profile, for example to modify BBB permeability. In 2000
and 2003, the VGCC channel activities of a series of PCU
1664 Current Medicinal Chemistry, 2013, Vol. 20, No. 13
derivatives of NGP1-01 were evaluated to determine struc-
ture-activity trends [26, 27]. These studies indicated that
both size and electrostatic properties were important deter-
mining factors for activity at the VGCC. Substitution on the
aromatic ring of the NGP1-01 structure could be used to in-
crease VGCC blocking activity, with meta-substituted com-
pounds being more active than ortho, and ortho substitution
more active than para substitution. Also, the introduction of a
methoxy instead of a nitro-group would generally lead to
increased activity. Notably, increasing the cage size (and
therefore the molecular volume) from a pentacyclic system
to a hexacyclic system, also led to an increase in activity.
These findings were also apparent in the QSAR studies per-
formed on the data set.
NH2
H Recently, PCU compounds have also been identified as
N
O been implicated in neurodegeneration [33]. Several of these
amantadine NGP1-01:
NMDA: 2.4 μM
NH2 sigma-1: 1.5 μM
VGCC: 86 μM
DAT: 55 μM
OH
memantine
N a non-selective adenosine receptor antagonist, and that sev-
OH aza-NGP1-01
N
TC-1:
F further be modified to create compounds that can act as both
Sigma-1: 10 nM
Sigma-2: 370 nM
Fig. (2). Structures of the polycyclic cage amines described in the
text. Detailed descriptions of the synthetic routes to NGP1-01 and
aza-NGP1-01 have been provided earlier [24, 26, 27].
The NMDAR activity was evaluated for NGP1-01 in two
studies. In a microdialysis study by Kiewert et al., NGP1-01
was shown to reach the CNS and block both the NMDAR
and VGCCs [23]. As noted for VGCC blocking activity (see
above) SAR studies of the PCU compounds at the NMDA
receptor also indicated that the size/volume of the PCU is
important for NMDA receptor blocking. NGP1-01 was able
to block the NMDAR in murine synaptoneurosomes with an
IC50 of 2.98 μM. This activity profile was similar to that of
memantine under similar experimental conditions (meman-
tine IC50 = 3.05 μM). Again, by increasing the ring size from
a 5- to a 6-membered ring, the activity was dramatically re-
duced (IC50 = 36 μM for the 6 membered NGP1-01 deriva-
tive). This finding that size is an important determining fac-
Geldenhuys and Schyf
tor for NMDAR blocking has been corroborated by another
study [28].
An early (1996) study has suggested that the combination
of NMDAR and VGCC blockers may have improved neuro-
protective potential in stroke models [22]. Based on this re-
port, NGP1-01 was evaluated in both a permanent and a
transient cerebral artery occlusion model of ischemic stroke.
In both these models, NGP1-01 (20 mg/kg) led to a signifi-
cant reduction in both the swelling (>80%) and infarct area
(>40%) of animals treated with NGP1-01 as compared to the
vehicle controls. Behaviorally, animals treated with NGP1-
01 performed significantly better than the vehicle control
groups [29, 30]. This neuroprotective activity of NGP1-01
may also in part be associated with its ability to also control
the entry of iron into cells through the VGCC 31, 32. By block-
ing the VGCC, NGP1-01 may also mitigate iron accumula-
tion following ischemic stroke [31, 32].
highly potent sigma receptor ligands, a receptor type that has
compounds have been shown to have nanomolar affinity for
the sigma-1 and sigma-2 receptor [34-40].
3. MAO-INHIBITION AND ADENOSINE A2 (A2A) RE-
CEPTOR ANTAGONISM
Converging lines of investigation have shown a benefi-
cial correlation between coffee drinkers and the decrease in
the incidence of Parkinson’s disease [41], an effect that has
since been attributed to the xanthine compound caffeine (Fig.
3). Pharmacological studies have determined that caffeine is
eral caffeine derivatives exhibit mild monoamine oxidase
(MAO) B inhibitory activity. Many compounds of dietary
origin, such as those containing the xanthine scaffold of caf-
feine, can easily be modified to design both A2A and MAO-B
inhibitors [42]. The MPTP model of parkinsonism was util-
ized to show that the protective activity of caffeine could be
reversed by the addition of an A2A agonist, suggesting that
caffeine derived its neuroprotective activity in part through
inhibition of the A2A receptor [43]. The xanthine scaffold can
A2A antagonists and MAO-B inhibitors [44]. MAO-B activ-
ity is important in the metabolism of dopamine, and inhibi-
tion of the activity of this enzyme leads to symptomatic re-
lief in Parkinson’s patients. The styrylxanthine or caffeinyl
scaffold of compounds such as CSC (Fig. 3) and istradefyl-
line (KW-6002; Fig. 3) can be modified to yield A2A antago-
nists [45] that also exhibit MAO-B inhibitory activity. A few
notable SAR trends can be seen for MAO-B inhibition by the
styrylxanthines [46-49]. Substitution on the aromatic styryl
ring plays in important role in MAO-B inhibition. Electron
withdrawing groups generally increase activity, with chlorine
substitution leading this trend. Also, the steric configuration
of the stilbene is important, since the trans form is favored to
the cis form, cis istradefylline being inactive. Selectivity and
activity for MAO-B and A2A can also be optimized by in-
creasing the linker size of the stilbene, as evidenced by a
series of (E,E)-phenylbutadiene caffeinyl derivatives. By
increasing the linker length, these caffeinyl derivatives were
seen to be very potent (MAO-B and A2A low nM potencies),
and the selectivity of MAO-B over MAO-A could be in-
Rationally Designed Multi-Targeted Agents Current Medicinal Chemistry, 2013, Vol. 20, No. 13 1665

creased [46-49]. The substrate pocket of MAO-A is smaller generative diseases such as cerebral stroke [52, 53] and Park-
than that of MAO-B, and the (E,E)-phenylbutadiene caf- inson’s disease [54-60] (Fig. 4). Thiazolidinedione (TZD;
feinyl derivatives appear to be too large for a facile fit into Fig. 4) compounds are PPAR-gamma agonists used in the
the substrate pocket of MAO-A. But these compounds fit treatment of type II diabetes [61]. Both pioglitazone and
comfortably into the substrate cavity of MAO-B after span- rosiglitazone have been shown to have protective effect in
ning the entrance cavity [50, 51]. pre-clinical models of Parkinson’s disease, including the
gold standard MPTP and 6-OH-dopamine models [55-59,
O CH3 62]. The neuroprotective activities of TZDs have been sug-
H3C N gested to be unrelated to their PPAR-gamma activity [63].
caffeine N
Of interest was the finding that pioglitazone was beneficial
O N against striatal toxicity of MPTP in a rodent model. We have
N
recently shown that this activity is related to the MAO in-
CH3
hibitory activity of TZDs, pioglitazone was found to be the
CH3 O most potent inhibitor of MAO-B, followed by rosiglitazone,
CH3
and then ciglitazone [64]. Crystallographic studies of piogli-
N
N O CH3
tazone and rosiglitazone in the MAO-B enzyme indicates
istradyfilline (KW-6002)
that the TZD head group binds in the substrate cavity facing
MAO-B: 28 μM O N N O the FAD catalytic site, while the remainder of the molecule
A2A: 2.2 nM CH3 resides in the entrance cavity. The TZD moiety is important
H3C for PPAR-gamma binding, with the tail end of pioglitazone
O
and rosiglitazone binding into a Y-shaped binding pocket
CH3
[65-67]. Taken together, selective compounds can be de-
H3C
N
N Cl signed to bind to MAO-B and PPAR-gamma, either indi-
CSC vidually or simultaneously as multi-targeted ligands.
O N N
MAO-B: 70 nM 5. MAO-B AND METAL CHELATION
CH3
A2A: 54 nM Metals have been suggested to play in important role in
neurodegeneration [32, 68-71]. These elements have the abil-
Fig. (3). Structures of caffeinyl compounds described in the text. ity to generate ROS species which induce cell death, and
may also play a role in the deleterious effects of misfolded
4. MAO-INHIBITION AND PPAR-GAMMA AGO- proteins such as amyloid-
in neurodegeneration [69]. For
NISM instance, significant accumulation of iron has been measured
in amyloid plaques as well as in neurofibrillary tangles in
Recently it was shown that peroxisome proliferator- Alzheimer’s disease patients [72]. Several groups have used
activated receptor gamma (PPAR-gamma) agonists such as metal chelation activity as starting point in designing multi-
pioglitazone and rosiglitazone are neuroprotective in several ple ligands. A series of compounds, also based on the stil-
neurodegenerative disease models, suggesting a promising bene scaffold were developed to interact with
-amyloid and
basis for the design of multiple ligands targeted at neurode- simultaneously exhibit metal chelating activity [73-75]. By

HO
O
N
CH3
CH3
NH
N N
S O
CH3
O
O O O
TZD

O O O

NH NH NH
S S S

O O O

troglitazone pioglitazone rosiglitazone

MAO-B: 2.07 μM 298 nM 832 nM

PPR-gamma: 780 nM 550 nM 76 nM

Fig. (4). Thiazolidinedione (TZD) anti-diabetic drugs.

1666 Current Medicinal Chemistry, 2013, Vol. 20, No. 13
chelating with metals in
-amyloid, the resulting ROS levels
should be reduced, with subsequent mitigation of protein
misfolding. A group of compounds was derived from
thioflavin T (ThT) and 6-iodo-2-(4'-dimethylamino)phenyl-
imidazo[1,2]pyridine (IMPY) [73-75] (Fig. 5). These com-
pounds served as lead structures for other compounds devel-
oped subsequently, and have an added bonus in that they are
brain permeable, which is of key importance for CNS drug
delivery [11]. Metal chelating moieties were introduced
based on the clioquinol template which utilizes the place-
ment of hydroxyl and nitrogen groups in close proximity as a
design for the chelation of metals. One of the compounds,
L2-b (Fig. 5), was found to chelate with metal ions with rank
potency of Cu2+/Zn2+>Fe2+>>Ca2+. L2-b as well as several of
the other derivatives tested, were able to prevent metal-
induced
-amyloid aggregation in a dose-dependent manner,
and also reduced the resultant ROS to afford protective ef-
fects in cell culture [76].
stilbene
N CH3
N Recently, a novel series of memoquin derivatives was cre-
I
N
CH3
IMPY
N activities to that of memoquin. These findings suggested that
NH
CH3
N
CH3
L2-b:
MAO-B: Ki = 28 nM
Chelation: 40 μM
Fig. (5). “Dual” metal chelating agents and MAO inhibitors.
H3C CH3
N
CH3
O O
N CH3
H3C
rivastigmine
Fig. (6). Combination of rivastigmine (an AChE inhibitor) and rasagiline (an MAO inhibitor) led to the development of ladostigil.
Geldenhuys and Schyf
Based on earlier findings that resveratrol, a stilbene, in-
hibits MAO, several stilbene-derived compounds identified
from the metal chelating studies were screened in an MAO-B
assay [77-79]. L2-b was found to be a facile MAO-B inhibi-
tor (IC50 = 52 μM), and the compounds tested showed pre-
ferred selectivity towards MAO-B compared to MAO-A
[80]. Substitution on the aromatic ring of the stilbene was
important for activity, with an increase in potency seen in a
series H<NH2<NMe2. L2-b was also shown to be a reversi-
ble MAO-B inhibitor.
6. AMYLOID-
AND CHOLINESTERASES
The polyamine-quinone memoquin has served as a tem-
plate structure for the development of molecules that prevent
both amyloid-
aggregation and act as acetylcholinesterase
(AChE) inhibitors [81-91]. Several molecular properties that
are relevant to the treatment and prevention of neuronal cell
death in Alzheimer’s disease are present in these compounds,
including the ability to interfere with the processing and ag-
gregation of amyloid-
peptides, the formation of reactive
oxygen species (ROS), and the activity of AChE. Memoquin
and several of its derivatives caused a significant decrease in
amyloid-
peptide- and ROS formation in animal models
[92-94]. Notably, in these studies, these compounds also
were reported to facilitate the reversal of behavioral deficits.
ated by linking the 2,5-diamino-benzoquinone core of these
compounds with motifs seen in known amyloid-
binding
agents, including curcumin, a natural product of dietary ori-
gin [92-94]. In these compounds, weaker AChE inhibitory
potencies were measured, while retaining equipotent anti-A

a balanced biological profile against the combined targets of
amyloid-
and AChE could be achieved.
7. MAO-B AND CHOLINESTERASE INHIBITION
One of the success stories in the development of multiple
targeted ligands is the development history of the multifunc-
tional ligand ladostigil (TV3326; Fig. 6). This compound is
currently in Phase II clinical trials (www.avphar.com) for the
treatment of mild cognitive impairment and comorbid de-
pression seen in Alzheimer’s and Parkinson’s disease. La-
CH
NH
rasagiline
O
H3C
N O CH
NH
CH3
ladostigil
Rationally Designed Multi-Targeted Agents
dostigil was designed based on the merger of two active mo-
lecular scaffolds that inhibit both AChE and MAO-B. Rivas-
tigmine (Fig. 6) is an AChE inhibitor which has been shown
to be beneficial in the treatment of both Alzheimer’s disease
and Parkinson’s disease [95, 96]. Rasagiline (Fig. 6) is an
irreversible MAO-B inhibitor that has been approved by the
FDA in 2006 for the treatment of Parkinson’s disease [97-
101]. Notably, rasagiline is one of few compounds that have
been suggested to possess disease-modifying activity in
Parkinson’s disease [102-104].
Ladostigil as a multiple targeted ligand has been created
through a combination of the rasagiline scaffold containing a
propargylamine moiety, and the carbamate moiety from the
structure of rivastigmine. This combination of activities af-
forded ladostigil the ability to be used for symptomatic re-
lief, as well as the potential to act as a disease-modifying
agent [104]. Specifically, the purported disease-modifying
ability of this molecule has been attributed to the propargy-
lamine moiety [105-109]. In addition, some studies have
suggested that the carbamate part of ladostigil is neuropro-
tective via heat shock protein (HSP) activity [110]. Notably,
ladostigil has been shown to have beneficial effects on cog-
nition [109]. In addition, upregulation of ERK 1/2 and PKC,
reduction of hydrogen peroxide, and prevention of the reduc-
tion in mitochondrial membrane potential all contribute to
the drug’s effects. It has also been shown to potentially act in
mitigating the progression of Alzheimer’s disease by increas-
ing the soluble fraction of amyloid precursor protein (APP)
[106, 111-114].
The coumarin- and chromone-derived structures (Fig. 7;
1 and 2) have been shown to be useful scaffolds that can
accommodate both MAO and AChE inhibitory activities. In
a recent study, several related compounds were shown to be
potent noncompetitive inhibitors of both enzymes [115].
SAR studies in the coumarin scaffold indicated a loss of ac-
tivity for AChE to be associated with meta and/or para sub-
stitution with a halogen moiety, while similar substitution
led to an increase in MAO-B inhibitory activity, similar to
that seen in the styrylxanthines [45].
CH3
CH3
H3C O O
1
O
O indanone scaffold was successfully employed to develop
O CH3
2:
MAO-B: pIC50 = 6.90
AChE: pKi = 4.53
Fig. (7). Coumarin derivatives that have been shown to serve as
templates for both MAO and AChE inhibitors.
The ability to inhibit both MAO and ChE enzymes was
also investigated in the development of another MTDD Alz-
Current Medicinal Chemistry, 2013, Vol. 20, No. 13 1667
heimer’s disease drug [116]. Here, the AChE inhibitor done-
pezil (Fig. 8) was used to connect the 1-benzylpiperidine
containing group with a moiety associated with MAO inhibi-
tion, N-[(5-benzyloxy-1-methyl-1H-indol-2-yl)methyl]-N-
methylprop-2-yn-1-amine (3). The moiety associated with
AChE inhibition binds into the catalytic cavity of the AChE
enzyme, whereas the moiety associated with MAO inhibition
binds into the substrate pocket of MAO. The most potent
compound (4) of the series was able to inhibit AChE (IC50 =
350 nM), BuChE (IC50 = 460 nM), MAO-A (IC50 = 5.2 nM)
and MAO-B (IC50 = 43 nM).
O
H3C
O
H3C
N
O
CH
N H3C
O N
N
CH3 4
CH
H3C
O N
N
CH3
3
Fig. (8). Structures of novel multi-targeted AChE- and MAO-B
inhibitors described in the text. Structure of donepezil shown at the
top of the figure.
The donepezil scaffold (Fig. 8) has also led to other multi
targeted drug design programs developing novel compounds
for Alzheimer’s disease treatment [117]. The benzylidene-
MAO inhibitors that also prevented amyloid-
aggregation,
metal chelation, and also possess antioxidant activity. These
compounds, in particular compound 5 (Fig. 9), inhibited
MAO-B at an IC50 = 7.5 μM, inhibited amyloid-
aggrega-
tion (at >80%), and was able to inhibit Cu-induced amyloid-

fibril formation [117].
8. HISTONE DEACETYLASE INHIBITORS AND PKC
ACTIVATORS
A novel emerging strategy for the treatment of Alz-
heimer’s disease is the combination of activators of protein
kinase C (PKC), and histone deacetylase (HDAC) inhibitors,
1668 Current Medicinal Chemistry, 2013, Vol. 20, No. 13 Geldenhuys and Schyf

as was recently described [118]. The basis for this strategy of we show that there is great promise in developing disease-
designed multiple ligand drugs was the expectation that PKC modifying agents based on the premise of targeting more
kinase activation would likely increase the alpha-secretase than one disease pathway known to be important in neurode-
pathway thereby increasing the soluble fraction of APP [119, generation. In support of the validity of this approach are the
120]. Additionally HDAC inhibition is thought to be poten- promising results emanating from early clinical studies of
tially neuroprotective via the blocking of oxidative stress ladostigil, which is now in clinical trials, and the emergence
[121]. A series of benzolactams was evaluated for both PKC – in preclinical stages of development – of several new com-
activation via soluble APP determination and HDAC inhibi- pounds described in this review.
tory activity via a homocysteine neuronal model of oxidative
stress. The findings indicated that the benzolactams tested CONFLICT OF INTEREST
were nanomolar inhibitors of PKC and were also able to The author(s) confirm that this article content has no con-
show neuroprotection via HDAC inhibition (10 – 20 μM). flict of interest.
Two compounds (one being compound 6; Fig 10) stood out
in that they were equipotent against HDAC and PKC (~10 ACKNOWLEDGEMENT
μM for both drug targets). This finding is significant consid-
ering that one of the major hurdles in multiple ligand design Declared none.
is the ability to align pharmacological potencies at the indi-
vidual drug targets within a similar range [122]. These ben- LIST OF ABBREVIATIONS
zolactams represent a novel group of compounds that could AChE = Acetylcholine esterase
effectively be developed into disease-modifying agents for
BBB = Blood-brain barrier
Alzheimer’s disease.
CNS = Central nervous system
HO
FDA = Federal Drug Administration of the United
States
HO
IMPY = 6-Iodo-2-(4'-dimethylamino)phenyl-
O
imidazo[1,2]pyridine

N
HSP = Heat shock protein
H3C MAO = Monoamine oxidase
CH3 MTDD = Multi-targeted designed drugs
5: NMDAR = N-methyl-D-aspartate receptor
MAO-B: IC50 = 7.50 μM PCU = Pentacycloundecyl
Anti-amyloid-beta: 80.1% at 20 μM PKC = Protein kinase C
ThT = Thioflavin T
Fig. (9). A multifunctional drug that inhibits MAO-B as well as
prevents amyloid-
aggregation. TZD = Thiazolidinedione

O VGCC = Voltage gated calcium channel

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Received: January 10, 2012 Revised: November 02, 2012 Accepted: November 02, 2012
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