Professional Documents
Culture Documents
Review
Journal of Pharmacy
And Pharmacology
REQUIMTE/Laboratrio de Farmacognosia, Departamento de Qumica, Faculdade de Farmcia, Universidade do Porto, Porto, Portugal and
Research Group on Quality, Safety and Bioactivity of Plant Foods, Department of Food Science and Technology, CEBAS (CSIC), Murcia, Spain
Keywords
biomedicinal chemistry; clinical efficacy of
natural products; drugs from natural sources;
structure/activity relationships
Correspondence
Paula B. Andrade, REQUIMTE/Laboratrio de
Farmacognosia, Departamento de Qumica,
Faculdade de Farmcia, Universidade do
Porto, Rua de Jorge Viterbo Ferreira, n 228,
4050-313 Porto, Portugal.
E-mail: pandrade@ff.up.pt
Received January 25, 2013
Accepted April 11, 2013
doi: 10.1111/jphp.12081
Abstract
Objectives Alzheimers disease (AD) is the most common cause of dementia,
being responsible for high healthcare costs and familial hardships. Despite the
efforts of researchers, no treatment able to delay or stop AD progress exists. Currently, the available treatments are only symptomatic, cholinesterase inhibitors
being the most widely used drugs. Here we describe several natural compounds
with anticholinesterase (acetylcholinesterase and butyrylcholinesterase) activity
and also some synthetic compounds whose structures are based on those of
natural compounds.
Key findings Galantamine and rivastigmine are two cholinesterase inhibitors
used in therapeutics: galantamine is a natural alkaloid that was extracted for the
first time from Galanthus nivalis L., while rivastigmine is a synthetic alkaloid, the
structure of which is modelled on that of natural physostigmine. Alkaloids include
a high number of compounds with anticholinesterases activity at the submicromolar range. Quinones and stilbenes are less well studied regarding cholinesterase
inhibition, although some of them, such as sargaquinoic acid or (+)-a-viniferin,
show promising activity. Among flavonoids, flavones and isoflavones are the most
potent compounds. Xanthones and monoterpenes are generally weak cholinesterase inhibitors.
Summary Nature is an almost endless source of bioactive compounds. Several
natural compounds have anticholinesterase activity and others can be used as
leader compounds for the synthesis of new drugs.
Introduction
Neurodegenerative diseases are increasing in developed
countries and they have in common the progressive and
specific loss of certain brain cell populations, leading to
functional or sensory dysfunction. Neurodegenerative disorders have some neuropathological hallmarks, such as
abnormal protein dynamics, bioenergetics and mitochondrial function impairment, oxidative stress and inflammation. In general neurodegeneration has a multifactorial
nature, being associated with environmental factors
and with genetic predisposition.[1] Neurodegeneration has
been the object of study of several teams with the intention
of finding a way to prevent, reduce or even treat
2013 Royal Pharmaceutical Society, Journal of Pharmacy and Pharmacology, 65, pp. 16811700
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Alzheimers disease
As mentioned above, the incidence of AD will increase as
the population ages, since age is the main risk factor for the
development of the disease.[4,6] However, epidemiological
findings associate several other factors with AD prevalence,
such as low education level, history of head trauma, highcaloric diet, folate-poor diet, sedentary lifestyle, low mental
ability in early life, and reduced mental and physical activity
during late life, as well as vascular disease risks (hypercholesterolaemia, hypertension, atherosclerosis, coronary heart
disease, smoking, obesity and diabetes).[7] Only a small percentage of AD cases is related to family history, whereby
specific genes are associated with increased susceptibility to
AD (amyloid precursor protein,[8] presenilin 1,[8] clusterin,[9]
sortilin-related receptor 1 gene[10] or apolipoprotein
allele E4).[8]
Symptomatically, AD is characterized by memory loss,
multiple cognitive impairment, disturbance of functions
such as language and executive function, and by several
neurobehavioural symptoms, including apathy, agitation
and anxiety.[11] Currently, AD cannot be diagnosed in the
absence of cognitive impairment/dementia[12] and a definitive diagnosis can only be established by post-mortem
examination of the AD patients brain.[4] Nevertheless,
research suggests that neurodegeneration starts 2030 years
before clinical symptoms appear.[13] It has recently been
reported that AD pathology can be detected pre-clinically
by using neuroimaging techniques and biomarkers.[12,14]
AD is characterized by the presence of neuritic plaques
and neurofibrillary tangles, mainly in the brain regions
responsible for learning, memory and emotional behaviour.[4] Neuritic plaques are microscopic extracellular
deposits of fibrils and amorphous aggregates of amyloid-b
peptide (Ab),[15] which result from an imbalance between
production and clearance of Ab in the brain.[16] Neurofibrillary tangles are intracellular fibrillar aggregates
of microtubule-associated hyperphosphorylated tau protein.[15] The reason for the appearance of neuritic plaques
and neurofibrillary tangles remains uncertain;[4] however,
the presence of these structures has several consequences in
affected brains. Ab accumulation might increase microglio1682
Cholinesterases
ACh is present throughout the nervous system, being essential for cerebral cortical development and activity, cerebral
blood flow control, sleepwake cycle and, mainly, for learning and memory processes.[18] Several studies in animal
models have shown the occurrence of learning or memory
deficits after anticholinergic treatment, highlighting the
importance of ACh for cognitive performance.[19,20] ACh is
stored in vesicles in the terminal nerves, being released
2013 Royal Pharmaceutical Society, Journal of Pharmacy and Pharmacology, 65, pp. 16811700
Acetylcholine
HO
OH
NH2
N
H
O
H
Glutamate
OH
H
N
O
O
OH
NH2
O
O
NH3
HO
NH2
NH3
O
NH2
N
+
N
+
Serine
NH2
O
HO
HO
Histidine
N
+
NH2
N
+
NH3
OH
O
HO
OH
NH2
N
H
O
O
O
O
NH3
OH
H
OH
NH2
HO
O
NH2
HO
N
+
NH2
NH3
O
O
Figure 1 Acetylcholine hydrolysis by acetylcholinesterase. Serine, histidine and glutamate constitute the catalytic triad in the esteratic site. Hydroxyl
group of serine induces a nucleophilic attack to acetylcholine, being stabilized by histidine and glutamate.
the carbonyl group of the ester substrate; (ii) histidine stabilizes the serine intermediate by strong hydrogen bonds;
(iii) the negative charge of glutamate stabilizes the histidinium cation.[25] The oxyanion hole contains hydrogen donors
which stabilize the tetrahedral intermediate of the substrate
that is formed during the catalytic process. The anionic
subsite (choline-binding subsite or hydrophobic subsite)
contains several aromatic residues, which are important for
the binding of quaternary ammonium ligands by p-cation
interactions.[25] However, the number of aromatic amino
acids differs according to the enzyme: some aromatic amino
acid residues present in the acyl pocket and in the peripheral site of AChE are replaced by aliphatic amino acids in
BuChE. As aliphatic amino acids are smaller than aromatic
amino acids, these alterations allow larger substrates to
enter the active site of BuChE. Thus, the active site of
BuChE can accommodate larger acyl groups, such as those
with four carbons (e.g. butyrylcholine) or aromatic rings
(e.g. benzoylcholine).[25]
Less information is available regarding BuChE, compared
with AChE, in the nervous system. However, it is known
that BuChE is expressed in a distinct population of neurons
and that it is important to cholinergic neurotransmission
regulation and to nervous system development.[22] In addition, the activity of BuChE is altered in patients with AD:
BuChE activity is reported to be significantly (4180%)
enhanced in brains of patients with advanced AD,[26] mainly
in regions affected by Ab plaques and neurofibrillary tangles.[22] In contrast, there is a deficit of 1060% of AChE in
affected brain regions of AD patients.[27] Furthermore, there
are several factors indicating that BuChE can be a potential
target in AD treatment: (i) BuChE has a higher half-life
2013 Royal Pharmaceutical Society, Journal of Pharmacy and Pharmacology, 65, pp. 16811700
1683
Cholinesterase inhibitors
Several new treatment options to halt the course of AD
have been studied, such as those that avoid Ab production
(preventing assembly of Ab monomers into potentially
cytotoxic oligomers; b- or g-secretase inhibition)[15] and
parental immunization with a synthetic human Ab peptide,
the latter of which has produced promising results.[29]
However, the treatments currently available are only symptomatic: memantine, which is an N-methyl-d-aspartate
(NMDA) receptor antagonist, and ChE inhibitors are the
only drugs available.[3,17] ChE inhibitors do not induce a
change in the natural course of AD, but temporarily mitigate some symptoms, since these drugs enhance the activation of synapses, thus improving cognition.[4,13] Other
approaches have been tried, with the aim of improving
cholinergic transmission, such as the increase of presynaptic ACh release or the stimulation of postsynaptic muscarinic or nicotinic receptors; these approaches have been
unsuccessful due to their lack of efficacy and unacceptable
side-effects.[17]
ChE inhibitors can be classified according to their selectivity for AChE or BuChE, or according to their mechanism
of action. Reversible inhibitors (e.g. tacrine or donepezil)
are competitively displaced from the active centre of
the enzyme by physiological ligands or other choline
esters. Pseudo-irreversible inhibitors (e.g. physostigmine
or rivastigmine) bind more firmly to the enzyme than a
physiological ligand. Irreversible inhibitors (e.g. organo1684
Alkaloids
Alkaloids constitute a wide family of compounds, which
generally have in common the presence of nitrogen atom(s)
in a cyclic ring. This is probably the largest group of
metabolites with ChE inhibitory activity at lower concentrations (Table 1). As described by Houghton and colleagues, the majority of alkaloids bind at the bottom of the
gorge of the active site, mainly at the oxyanion hole area, via
the positively charged nitrogen.[25] The first known AChE
inhibitor was physostigmine (Figure 2), an alkaloid isolated
for the first time in 1864 from Physostigmina venenosum
Balf., which was used in therapy before the discovery of
ACh as neurotransmitter.[76] However, physostigmine is
quite polar, being distributed throughout the body, and
only a small amount reaches the central nervous system.
Physostigmine inhibits both AChE and BuChE in a similar
submicromolar range (concentration required to inhibit
50% of the enzyme ( IC50) of 0.015 and 0.016 mm, respectively) (Table 1).[43,44] The carbamate position is essential for
the activity of physostigmine, because when the ester bond
of physostigmine is hydrolysed, forming eseroline, the
inhibitory activity is not observed. Furthermore, it is known
2013 Royal Pharmaceutical Society, Journal of Pharmacy and Pharmacology, 65, pp. 16811700
Table 1
Compounds
Alkaloids
Axillaridine A
Berberine
a-Chaconine
Cymserine
Galantamine
Groenlandicine
Huperzine A
Infractopicrin
Juliflorine
Neostigmine
Petrosamine
Physostigmine
Pyridostigmine
Rivastigmine
Sarsalignenone
Sarsalignone
a-Solanine
Tubocurarine
Coumarins
Decursin
Decursinol
Heraclenol-2-O-angelate
Imperatorin
Isopimpinellin
Marmesin
Methoxsalen
Scopoletin
Flavonoids
Apigenin
Kaempferol
Leufolin A
Leufolin B
Luteolin
Mimulone
Naringenin
Pomiferin
Quercetin
Rutin
Tamarixetin
Quinones
Mansonone E
Sargaquinoic acid
Stilbenes
(+)-a-Viniferin
Gnetol
Kobophenol A
Resveratrol
Terpenes
1,8-Cineole
2-Carene
3-Carene
Cryptotanshinone
Dihydrotanshinone
Leoheteronin A
IC50 (mM)
5.21
0.44
17
0.10
4.00
0.54
0.082
9.72
0.42
0.036
0.091
0.015
0.091
501
5.83
7.02
14
63
3900
28
>1000
165
0.32
67
3.07
52
21.5
30.4
Butyrylcholinesterase
Source
Reference
IC50 (mM)
Source
Reference
Electric eel
Electric eel
Human recombinant
Human erythrocytes
Electric eel
Electric eel
Rat cortex
Bovine erythrocytes
Electric eel
Guinea-pig ileum
Pacific electric ray
Human brain
Mouse brain
Electric eel
Electric eel
Electric eel
Human recombinant
Human erythrocytes
[33]
2.49
3.44
0.066
0.001
7.96
3.32
74.4
>100
0.12
0.19
N.F.
0.016
0.30
19.95
4.29
2.18
0.17
350
Horse serum
Horse serum
Human serum
Human erythrocytes
Equine serum
Horse serum
Rat serum
Equine serum
Horse serum
Guinea-pig ileum
[33]
Human plasma
Human plasma
Equine serum
Horse serum
Horse serum
Human serum
Human plasma
[44]
N.M.
N.M.
Electric eel
Electric eel
Fly lysate
N.M.
Rat cell line
N.M.
[48]
Equine serum
Equine serum
[49]
Rat brain
N.M.
[53]
N.M.
N.M.
N.M.
[54]
Equine serum
Equine serum
Horse serum
Horse serum
Horse serum
N.M.
[56]
Horse serum
N.M.
[60]
Horse serum
[63]
Horse serum
[63]
Human serum
Human
Human
[66]
[34]
[35]
[36]
[37]
[34]
[38]
[39]
[40]
[41]
[42]
[43]
[45]
[37]
[33]
[33]
[35]
[47]
[48]
[49]
[49]
[50]
[48]
[51]
[52]
[54]
N.F.
N.F.
7.5
14.4
N.F.
N.F.
N.F.
N.F.
N.F.
62.5
1.6
3.6
N.F.
20.6
1494
N.F.
420.8
44.6
160.6
[53]
Electric eel
N.M.
[60]
2.0
N.M.
[62]
N.F.
N.F.
115.8
>500
N.M.
N.M.
[62]
1.3
N.F.
15.9
Bovine erythrocytes
Bovine erythrocytes
Bovine erythrocytes
Recombinant human
Recombinant human
Mouse brain
[64]
23.5
23.3
670
900
200
4.67
0.89
11.6
[56]
[56]
[57]
[58]
[59]
[54]
[61]
[62]
[65]
[65]
[67]
[67]
[68]
2013 Royal Pharmaceutical Society, Journal of Pharmacy and Pharmacology, 65, pp. 16811700
[35]
[36]
[37]
[34]
[38]
[39]
[40]
[41]
[46]
[37]
[33]
[33]
[35]
[47]
N.F.
N.F.
Rat brain
Human erythrocytes
Human erythrocytes
Electric eel
Pacific electric ray
Electric eel
N.M.
15.6
91.5
2045
96
353.9
28.2
22.3
[34]
62.4
0.026
N.F.
N.F.
2000
6.66
5.51
N.F.
[49]
[55]
[55]
[56]
[57]
[58]
[59]
[54]
[61]
[67]
[67]
1685
Table 1
(Continued)
Acetylcholinesterase
Compounds
IC50 (mM)
Leopersin G
Limbatolide A
Limbatolide B
Limbatolide C
Myrtenal
Taraxerol
Ursolic acid
a-Pinene
Xanthones
Allanxanthone A
Triptexanthoside C
Others
Arisugacin A
Territrem B
12.9
38.5
47.2
103.7
170
98.4
93.8
630
95
13.8
0.001
0.076
Butyrylcholinesterase
Source
Reference
IC50 (mM)
Mouse brain
Electric eel
Electric eel
Electric eel
N.M.
N.M.
Electric eel
Bovine erythrocytes
[68]
N.F.
22.3
17.5
14.2
N.F.
N.F.
41.1
N.F.
Electric eel
N.M.
[72]
N.M.
N.M.
[74,75]
[69]
[69]
[69]
[70]
[71]
[59]
[64]
[73]
[75]
19.1
N.F.
Source
Reference
Horse serum
Horse serum
Horse serum
[69]
Horse serum
[59]
Horse serum
[72]
[69]
[69]
N.F.
N.F.
H
O
N
O
N
+
N+
Physostigmine
Pyridostigmine
Neostigmine
Rivastigmine
N
O
H
O
Cymserine
N
2013 Royal Pharmaceutical Society, Journal of Pharmacy and Pharmacology, 65, pp. 16811700
(a) Isoquinoline
(b) Indole
OH
O
O
+
N
N
H
N+
O
+
N
OH
O
Tubocurarine
(c) Terpenic
OH
Berberine
N+
Galantamine
Geissospermine
(d) Acridine
Infractopicrin
Angustine
(e) Piperidinium
Br
HO
N
O
HO
NH2
+
N
B
N
Huperzine A
Petrosamine
Juliflorine
(f) Steroidal
(g) Lycorine
H
H
N
H
H
-Solanine
-Chaconine
O
OH
O
O
O
HO
H
OH
HO
OH
R=
OH
HO
OH
O
OH OH
HO
OH
OH
OH
H
O
OH
HO
RO
R=
N
H
N
H
Axillaridine A
H
N
Sarsalignone
O
HO
OH
O
Sarsalignenone
OH
Figure 3 Chemical structure of alkaloids from different subgroups. (a) Isoquinoline: berberine, tubocurarine and galantamine. (b) Indole: geissospermine, infractopicrin and angustine. (c) Terpenic: huperzine A. (d) Acridine: petrosamine. (e) Piperidinium: juliflorine. (f) Steroidal: a-solanine,
a-chaconine, axillaridine A, sarsalignone and sarsalignenone. (g) Lycorine.
Galantamine (Figure 3a) is a natural alkaloid with an isoquinoline skeleton that was isolated for the first time from
Galanthus nivalis L.[76] Galantamine selectively and reversibly inhibits AChE, increases presynaptic ACh release and
postsynaptic neurotransmission, by acting as an allosteric
ligand at nicotinic receptors.[17] Galantamine is well tolerated and is being used in AD therapeutics. A recent work
concluded that long-term galantamine treatment allows
cognitive and global stabilization of AD.[84] Galantamine
displays fairly strong in-vitro AChE inhibitory activity
(IC50 = 4.0 mm) (Table 1).[37] The increase in the size of the
alkyl group linked to the nitrogen of galantamine seems to
improve the AChE inhibitory activity: N-(14-methylallyl)
norgalantamine (IC50 = 0.16 mm) > N-allylnorgalantamine
(IC50 = 0.18 mm) > galantamine. N-(14-Methylallyl)norgalantamine and N-allylnorgalantamine are derived from the
waste generated by the industrial production of galantamine from Leucojum aestivum L. leaves. The application
of these compounds in therapeutics would allow efficient
use of those wastes.[85] In contrast, the loss of the N-methyl
group of galantamine, such as in epinorgalantamine, is
2013 Royal Pharmaceutical Society, Journal of Pharmacy and Pharmacology, 65, pp. 16811700
1687
2013 Royal Pharmaceutical Society, Journal of Pharmacy and Pharmacology, 65, pp. 16811700
Furanocoumarins
O
OH
O
O
OH
Scopoletin
Marmesin
R2
O
Pyranocoumarins
O
R1
Compound
R1
R2
OH
O
Compound
Decursinol
OH
Heraclenol-2-O-angelate
OH
Imperatorin
Isopimpinellin
OCH3
OCH3
Methoxsalen
OCH3
Decursin
Figure 4 Chemical structure of coumarins: scopoletin, furanocoumarins (heraclenol-2-O-angelate; imperatorin, isopimpinellin, marmesin and
methoxsalen) and pyranocoumarins (decursinol and decursin).
Coumarins
Coumarins are benzo-a-pyrones (a benzene ring joined
to a pyrone ring) with interesting pharmacological proper-
ties.[100] The natural compound scopoletin (Figure 4) inhibits AChE with an IC50 of 52 mm (Table 1).[52] This has been
confirmed by in-vivo assays, in which scopoletin and its
glucoside scopolin increased rat brain extracellular ACh
concentration.[101]
Methoxsalen (Figure 4), a furanocoumarin also known as
xanthotoxin, was extracted from Poncirus trifoliata (L.) Raf.
and has strong AChE inhibitory activity (IC50 = 3.07 mm)
(Table 1). The anti-AChE activity of methoxsalen was confirmed by the inhibition of mouse brain enzyme and amelioration of drug-induced behavioural impairment in an
AD-like mouse model.[51,102] Two furanocoumarins with
good anti-AChE activity were extracted from Angelica acutiloba (Siebold & Zucc.) Kitag: methoxsalen and isopimpinellin (IC50 = 0.32 mm) (Figure 4, Table 1).[50] Imperatorin (a
prenylated furanocoumarin) and heraclenol-2-O-angelate
(Figure 4) were isolated from Angelica archangelica L. and
both compounds showed a good BuChE inhibitory activity
(IC50 of 14.4 and 7.5 mm, respectively) (Table 1). The
2013 Royal Pharmaceutical Society, Journal of Pharmacy and Pharmacology, 65, pp. 16811700
1689
authors concluded that C8 and C5 substituted furanocoumarins are BuChE inhibitors.[49] Other authors argue that
coumarins (e.g. methoxsalen, imperatorin and bergapten)
are more selective against BuChE than against AChE and
that the difference in enzymatic inhibition can reach
50%.[49,103] In contrast to the negative effect of glycosylation
on anti-ChE activity, marmesin (Figure 4) (a furanocoumarin) and its corresponding glycoside have similar activity
(IC50 of 67 and 68 mm, respectively).[48]
Kang et al. showed that the compound isolated from
Angelica gigas Nakai with high activity against AChE was a
pyranocoumarin, decursinol (Figure 4), and that the
inclusion of a prenyl group instead of hydroxyl group at
C3, such as in decursin (Figure 4), induces a decrease of
activity (Table 1).[48] Furthermore, decursin ameliorated
scopolamine-induced amnesia in mice, inhibiting the hippocampus AChE activity by 34%.[104]
Besides the study of the anti-ChE effects of natural coumarins, researchers have been using the coumarin skeleton
to produce compounds with stronger activity. Catto and
collaborators designed a series of coumarin alkylamides
with the structural determinants of donepezil, a known
synthetic non-competitive and reversible AChE inhibitor.
Thus, 6,7-dimethoxycoumarin, carrying a protonable benzylamino group linked to C3 by specific linkers, showed a
good activity towards both AChE and BuChE. Apparently,
these compounds are able to bind to both catalytic and
peripheral binding sites of ChE, the activity being highly
dependent on the length of the spacer and on the methoxyl
substituents.[105] Coumarin derivatives with a N-benzyl
pyridinium moiety also have two AChE binding sites,
and are active at the nanomolar range.[106] In addition,
3-thiadiazolyl and thioxo-1,2,4-triazolylcoumarin derivatives have very good activity also at nanomolar concentrations against both AChE and BuChE.[107]
Flavonoids
Flavonoids constitute a class of polyphenols characterized
by a diphenylpropane (C6-C3-C6) skeleton, which consists of
two aromatic rings, each bearing at least one aromatic
hydroxyl, connected by a carbon bridge, forming (or not) a
third ring. Flavonoids are divided into subclasses based on
the connection of the two aromatic rings, degree of oxidation and also the functional groups of the third ring
(flavanols, flavanones, anthocyanidins, flavones, flavonols,
isoflavones, flavan-3-ols, flavanonols, aurones or chalcones). The majority of flavonoids naturally occur as glycosides or other conjugates, which explains the great variety of
compounds, and are considered to be the most relevant
class of phenolic compounds.[108]
Uriarte-Pueyo and collaborators reviewed the ChE
inhibitory capacity of 128 flavonoids, concluding that the
1690
compounds with higher AChE activity were flavones or isoflavones. Thus, the carbonyl group at C4 seems to be important to anti-AChE activity. This team verified that among
the studied flavonoids, the most potent one was a synthetic
compound, 6,7-dimethoxy-3-[4-(pyrrolidin-1-ylmethyl)phenyl]-4H-chromen-4-one (IC50 = 0.004 mm), followed by
6,7-dimethoxy-2-[4-(piperidin-1-ylmethyl)-phenyl]-4Hchromen-4-one (IC50 = 0.034 mm) and 6,7-dimethoxy-2-[4(pyrrolidin-1-ylmethyl)-phenyl]-4H-chromen-4-one
(IC50 = 0.093 mm). The substituents pyrrolidin-1-ylmethyl
or piperidin-1-ylmethyl at C4, as well as methoxyl groups
at C6 or C7, appear to play an important role in the AChE
inhibitory activity of flavonoids.[109] The compounds containing a flavonoid moiety and a second moiety, which
could be amino alkyl, pyrrolidine or piperidine through an
appropriate spacer oxygen atom or alkoxyl group (O-CH2),
are able to interact with both the peripheral and the catalytic site of AChE: the flavonoid moiety is able to interact
with the peripheral site, while the second moiety interacts
with the catalytic site.[110] Compounds with pyrrolidine
or piperidine groups exhibited higher activity than those
with amino alkyl groups, indicating the key role of a
conformation-constrained hydrophobic moiety for AChE
inhibition.[110]
Among natural flavonoids, pomiferin (Figure 5), a
prenylated isoflavone extracted from Maclura pominifera
(Raf.) C.K. Schneid., was found to possess high anti-AChE
activity (IC50 = 96 mm) (Table 1). In contrast, flavanones
were less active against AChE than flavones, which suggested that the double bound at C2-C3 is important for the
anti-AChE activity.[109] Additionally, 5-geranyl-5,7,2,4tetrahydroxyflavone was the most potent AChE and BuChE
inhibitor isolated from Morus lhou Koidz., highlighting the
importance of the flavone skeleton. This compound, as well
as others not substituted at C3, are mixed inhibitors, while
the C3-prenyl substituted flavones are non-competitive
inhibitors, indicating that the presence of a prenyl group at
C3 affects the interactions with the enzyme.[111]
Considering natural and common flavonoids, fustin
(Figure 5) inhibited AChE (no IC50 value available) and
decreased the expression of the gene encoding AChE by
Ab.[112] Naringenin (Figure 5), a flavanone from Citrus junos
Siebold ex Tanaka, ameliorated scopolamine-induced
amnesia in mice, also having AChE inhibitory activity in
vitro, albeit at high concentrations (Table 1).[113] For prenylated flavonoids, the presence of a geranyl group at C6 is
important for anti-ChE activity, as demonstrated when
mimulone (IC50 = 91.5 mm) was compared with naringenin
(IC50 = 2045 mm) (Figure 5, Table 1).[56]
Several works have reported the strong anti-ChE activity
of galangin (a flavonol) compared with other flavonoids
tested under the same conditions.[53,114] Galangin was
reported to be the most potent BuChE inhibitor among
2013 Royal Pharmaceutical Society, Journal of Pharmacy and Pharmacology, 65, pp. 16811700
R5
R4
R6
R5
R1
R1
R6
R2
R4
R3
R3
R2
Compound
R1
R2
R3
R4
R5
R6
Compound
R1
R2
R3
R4
R5
R6
Fustin
OH
OH
OH
OH
Fisetin
OH
OH
OH
OH
Mimulone
OH
OH
OH
Galangin
OH
OH
OH
Naringenin
OH
OH
OH
Kaempferol OH
OH
OH
OH
OH
HO
O
OH
O
HO
OH
Leofolin A
Myricetin
OH
OH
OH
OH
OH
OH
Quercetin
OH
OH
OH
OH
OH
Rutin
OH
OH
OH
Tamarixetin OH
OH
OH
OH
OCH3
Apigenin
OH
OH
OH
Luteolin
OH
OH
OH
OH
Rutinoside OH
O
HO
OH
HO
O
OH
OH
O
O
O
Leofolin B
HO
OH
O
OH
Pomiferin
HO
OH
Figure 5 Chemical structure of flavonoids from different subgroups: flavanonols (fustin), flavanones (mimulone, naringenin and leufolin A); flavones (apigenin, leofolin B and luteolin); flavonols (fisetin, galangin, kaempferol, myricetin, quercetin, rutin and tamarixetin) and prenylated isoflavones (pomiferin).
2013 Royal Pharmaceutical Society, Journal of Pharmacy and Pharmacology, 65, pp. 16811700
1691
Benzoquinones
OH
Sargaquinoic acid
O
Naphthoquinones
O
O
O
Compound
Compound
Mansonone C
Mansonone E
Mansonone G
OH
Mansonone H
OH
R
O
Figure 6
Chemical structure of quinones: benzoquinone (sargaquinoic acid) and naphthoquinones (mansonones C, G, E and H).
Quinones
Quinones are widespread in nature, being essential for
many biochemical processes, such as mitochondrial respiration or photosynthesis. Quinones also have an important
role in an organisms defence, being able to inhibit bacterial, fungal and parasitic growth. All these properties are
related to the quinonoid structure, which has a pro-oxidant
character. Quinones may undergo redox cycling catalysed
by flavoenzymes, generating semiquinones or hydroquinones, which can create reactive oxygen species or react
with nucleophiles.[117] For all these reasons, it is not surprising that quinones are able to inhibit several proteins, such
as topoisomerase[118] or RNA polymerase.[119] However, there
are not many studies concerning the inhibition of ChE by
quinones.
The quinonoid group seems to be important for AChE
inhibition, since dopamine autoxidation can inactivate
AChE, mainly by direct interaction of quinone or semiquinone oxidation products with the enzyme.[120] Sargaquinoic
acid (Figure 6) is a natural benzoquinone extracted from
1692
Stilbenes
Stilbenes are a small family of secondary metabolites
derived from the phenylpropanoid pathway. Resveratrol
(Figure 7) is probably the most extensively studied stilbene
and has potent anti-cancer, anti-inflammatory and antioxi-
2013 Royal Pharmaceutical Society, Journal of Pharmacy and Pharmacology, 65, pp. 16811700
OH
HO
HO
HO
OH
HO
HO
Resveratrol
Gnetol
OH
HO
HO
HO
O
OH
OH
HO
OH
OH
OH
HO
O
O
OH
OH
HO
HO
Figure 7
(+)--Viniferin
HO
Kobophenol A
Chemical structure of stilbenes: monomers (resveratrol and gnetol) and oligomers ((+)-a-viniferin and kobophenol A).
dant properties.[122] In spite of the few studies about antiChE activity of stilbenes, some compounds of this class
with anti-ChE activity are currently known. Gnetol
(Figure 7), a stilbene isolated from Ficus foveolata Pittier,
has good activity towards BuChE (Table 1), via a reversible
and competitive mechanism.[63] (+)-a-Viniferin (Figure 7)
inhibits AChE in a dose-dependent manner (IC50 =
2.0 mm) (Table 1), by a non-competitive mechanism. (+)-aViniferin is a trimer of resveratrol, which itself has only a
weak effect on AChE, since it does not provide 50% inhibition at a concentration of 500 mm (Table 1). Thus, stilbeneoligomerization favours AChE inhibition. Nevertheless,
kobophenol A (Figure 7) (IC50 = 115.8 mm), a tetramer of
resveratrol, has lower AChE inhibitory potency than (+)-aviniferin (Table 1), probably due to steric hindrance.[62]
Other stilbenes with reported AChE inhibitory activity
are trans-3,5-dimethoxystilbene (IC50 = 9 mm), trans-3,5dimethoxy-2-prenylstilbene (IC50 = 19 mm) and furanokurzin (IC50 = 42 mm).[123]
Terpenic compounds
Terpenes, which are built up from isoprene subunits, constitute the most numerous and structurally diverse group of
2013 Royal Pharmaceutical Society, Journal of Pharmacy and Pharmacology, 65, pp. 16811700
1693
(a)-Monoterpenes
(b)-Diterpenes
O
O
O
O
O
1,8-Cineole 3-Carene
2-Carene
Myrtenal
-Pinene
Cryptotanshinone
Dihydrotanshinone
OH
O
O
OH
Leoheteronin A
Leopersin G
O
O
HO
HO
Leucisterol
Taraxerol
O
O
Limbatolide A
OH
Compound
OH
OH
OH
Limbatolide B OCH3
Limbatolide C
R
H
O
OH
HO
HO
Ursolic acid
Oleanolic acid
Sclareol
Figure 8 Chemical structure of terpenic compounds: (a) monoterpenes: 1,8-cineole, 3-carene, 2-carene, myrtenal and a-pinene; (b) diterpenes:
cryptotanshinone, dihydrotanshinone, leoheteronin A, leopersin G, limbatolide A, B and C and sclareol; (c) triterpenes and steroids: leucisterol,
taraxerol, ursolic acid and oleanolic acid.
2013 Royal Pharmaceutical Society, Journal of Pharmacy and Pharmacology, 65, pp. 16811700
OH
OH
OH
OH
OH
OH
O
O
O
OH
HO
OH
OH
O
OH
Allanxanthone A
Figure 9
Bellidifolin
OH
OH
Triptexanthoside C
cluded that the methoxyl group at C15 favours AChE inhibition. With respect to BuChE, the three mentioned
compounds have similar potency (IC50 = 14.222.3 mm)
(Table 1).[69] Sclareol (Figure 8b) is a diterpenoid extracted
from Salvia chrysophylla Stapf., and is reported to have good
activity against both AChE and BuChE.[128] Labdane-type
diterpenoids, like leoheteronin A (with a 8,13 diene and
15,16-epoxy group) and leopersin G (with a 13-diene and a
15,16 epoxy group) (Figure 8b), are also promising AChE
inhibitors (Table 1).[68]
Ursolic acid, taraxerol, leucisterol and oleanolic acid
(Figure 8c) are among the triterpenes and steroids with
anti-ChE activity described in literature.[129] ulhaoglu and
collaborators reported that ursolic and oleanolic acids are
selective against AChE.[128] Other researchers showed that
the IC50 towards BuChE of ursolic acid is lower than that
against AChE (Table 1).[59] Ursolic acid inhibits AChE in a
competitive/non-competitive way,[130] although Fatima et al.
did not verify any activity against AChE or BuChE.[131] Leucisterol is more potent against BuChE (IC50 = 3.2 mm) than
against AChE (IC50 = 83.6 mm).[131] Taraxerol inhibits AChE
in a dose-dependent manner and its activity is similar to
that of ursolic acid (Table 1).[52] Withanolides are C28steroidal lactone triterpenoids that can be isolated from
several species, such as Withania somnifera (L.) Dunal or
Ajuga bracteosa Wall. ex Benth. They inhibit both AChE
(IC50 = 2030 mm) and BuChE (IC50 = 5085 mm).[132]
Xanthones
Xanthones are secondary metabolites produced by plants,
fungi and lichens, important for providing protection
against insects, plant viruses, bacterial infections and animal
predation.[133] In spite of having only one additional
benzene ring than coumarins, xanthones are, in general,
weak ChE inhibitors, whereby the xanthone ring (dibenzog-pyrone) by itself does not appear to automatically confer
activity.[25,73,134] However, the presence of a lipophilic side
Others
Here we consider some compounds that due to their features were not included in any of the previous sections, but
deserve to be mentioned.
Territrem B (Figure 10) is a fungal metabolite isolated
from Aspergillus terreus Thom and has high AChE inhibitory activity (IC50 = 0.076 mm), acting by a mechanism different from those of other AChE inhibitors: territrem B
blocks the entrance of ACh into AChE by hydrophobic
interaction with the lipophilic amino acid in the entry of
the binding site channel. Several authors report that this
binding is kinetically irreversible, at least within the duration of study. In contrast to other inhibitors, territrem B
shows a one-to-one stoichiometry and appears not to bind
to subsites but to all the active gorge.[75,136138] The substantial and innovatory activity of territrem B encouraged the
search for new territrem B derivatives. Preliminary
structureactivity investigations suggested that the 2-en-1one moiety and the planar conformation were essential for
the AChE inhibitory activity.[138,139] No BuChE inhibition
has been observed with territrem B.[137]
Arisugacins are meroterpenoids from Penicillium species
with good anti-ChE activity described. Arisugacins A and
B (Figure 10) are selective inhibitors of AChE in vitro
2013 Royal Pharmaceutical Society, Journal of Pharmacy and Pharmacology, 65, pp. 16811700
1695
OH
O
OH
O
R1
O
O
R2
Figure 10
Compound
R1
R2
Arisugacin A
OCH3
Arisugacin B
Territrem B
OCH3
OCH3
Declarations
(Table 1).[140] As arisugacin A does not contain nitrogen, it is
suggested that the binding to AChE is by an electrondonatingelectron-withdrawing interaction. Arisugacin A
binds to peripheral anionic site by its dimethoxyaryl group,
whereby the dimethoxyaryl group stays positioned in the
opening of the catalytic gorge.[74] Similarly to territrem B,
arisugacin is highly selective towards AChE rather than
BuChE.[74]
Conclusions
AD is a neurodegenerative disease currently without effective treatment. ChE inhibitors can mitigate symptoms,
improving cognitive function due to the increase of ACh
half-life. Inhibitors of both AChE and BuChE could have
some advantages relative to compounds selective towards
AChE, as BuChE activity is normally enhanced in AD
patients brains. Nature is a source of new compounds and a
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Conflict of interest
The Author(s) declare(s) that they have no conflicts of
interest to disclose.
Funding
The authors are grateful to Fundao para a Cincia e a Tecnologia (FCT) for grant no. PEst-C/EQB/LA0006/2011, to
Consolider Ingenio 2010 Project CSD2007-00063 FUN-CFOOD, and to Grupo de Excelencia de la Rgion de Murcia
04486/GERM/06. B. Pinho thanks FCT for the grant
(SFRH/BD/63852/2009). Part of this work was carried out
under international cooperation within the CYTED Programme, Thematic Network Action 112RT0460 Characterization and evaluation of functionality and safety of
bioactive compounds from Iberian-American fruits for
food ingredients (CORNUCOPIA).
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