molecules

Review
Antiviral Activities of Oleanolic Acid and
Its Analogues
Vuyolwethu Khwaza, Opeoluwa O. Oyedeji and Blessing A. Aderibigbe *
Department of Chemistry, University of Fort Hare, Alice Campus, Alice 5700, Eastern Cape, South Africa;
vuyolwethukhwaza@gmail.com (V.K.); ooyedeji@ufh.ac.za (O.O.O)
* Correspondence: blessingaderibigbe@gmail.com; Tel.: +27-40-602-2266; Fax: +08-6730-1846
Academic Editors: Patrizia Ciminiello, Alfonso Mangoni, Marialuisa Menna and



Orazio Taglialatela-Scafati

Received: 27 July 2018; Accepted: 5 September 2018; Published: 9 September 2018

Abstract: Viral diseases, such as human immune deficiency virus (HIV), influenza, hepatitis, and
herpes, are the leading causes of human death in the world. The shortage of effective vaccines or
therapeutics for the prevention and treatment of the numerous viral infections, and the great increase
in the number of new drug-resistant viruses, indicate that there is a great need for the development
of novel and potent antiviral drugs. Natural products are one of the most valuable sources for drug
discovery. Most natural triterpenoids, such as oleanolic acid (OA), possess notable antiviral activity.
Therefore, it is important to validate how plant isolates, such as OA and its analogues, can improve
and produce potent drugs for the treatment of viral disease. This article reports a review of the
analogues of oleanolic acid and their selected pathogenic antiviral activities, which include HIV, the
influenza virus, hepatitis B and C viruses, and herpes viruses.

Keywords: HIV; influenza virus; HBV/HCV; natural product; triterpenoids; medicinal plant

1. Introduction
Viral diseases remain a major problem for humankind. It has been reported in some reviews that
there is an increase in the number of viral diseases responsible for death and morbidity around the
world [1,2]. According to the recent release from the National Health Laboratory Service (NHLS),
influenza kills approximately 6000–11,000 South Africans every year [3]. Although other treatments
eradicate some pathogens, such as polio, mumps and smallpox, other viral diseases, such as the
hepatitis C virus (HCV) and the human immunodeficiency virus (HIV), have proven difficult to
combat using the conventional treatment approach. In addition, the increase in viral resistance to
drugs, as well as the serious adverse effects of antiviral drugs, results in serious medical problems,
particularly when drugs are administered in combination over a prolonged treatment period [4].
Thus, there is a great need to develop novel potential antiviral agents from different sources, such as
medicinal plants or natural products, which have been used in many regions as antiviral agents for
many years [5–7].
Natural products have been proven as the main source of biologically active compounds, and they
are potentially useful for drug development [8]. Pentacyclic triterpenes (PTs) are the most significant
group of phytochemicals synthesized from plants through cyclization of squalene, and are known
as a large class of secondary plant metabolites that are constructed by isoprene (2-methylbutadiene)
(C5H8) units [9]. Structurally, they contain 5- and 6-membered rings (A, B, C, D, and E). The carbon
skeleton of PTs is divided into six different subgroups: Oleanane (1), ursane (2), friedelane (3),
hopane (4), lupine (5), and gammacerane (6) (Figure 1) [10]. It has been reported that approximately
20,000 triterpenoids exist in nature [11,12].

Molecules 2018, 23, 2300; doi:10.3390/molecules23092300 www.mdpi.com/journal/molecules

Molecules 2018, 23, 2300 2 of 14
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E E E
E E E
C D C D C D
C D C D C D
A B A B A B
A B A B A B
1 2
1 3
2 3

E E
E
E E
C C E C
D D D
C C D C
A B D D
A B A B
A B B
A B A
4 5 6
4 5 6
Figure1.1.Types
Figure Typesof
ofpentacyclic triterpenesstructures.
pentacyclic triterpenes structures.
Figure 1. Types of pentacyclic triterpenes structures.
Antivirals
Antivirals arearecompounds
compoundswhich whichprevent
prevent viral
viral development.
development. Some Someofofthese these antivirals
antivirals cancanbe be
isolated Antivirals
from
isolated from are
sources, compounds
sources,such which
suchasasplants, prevent
plants,animals, viral
animals, bacteria development.
bacteria or or fungi, Some
fungi,while of
whileothers these
otherscan antivirals
canbebe can
obtained
obtained be
by by
isolated
chemical
chemical from
synthesis sources,
synthesis [13,14]. such
[13,14]. as plants,
Antiviral
Antiviral animals, bacteria
mechanisms
mechanisms identified
identifiedor from
fungi, while products
fromnatural
natural others
productscan be shed
have
have obtained
shed by
light
light on on
howchemical
they
how they synthesis
interact
interact with [13,14].
thethe
with Antiviral
viral lifelife
viral mechanisms
cycle,
cycle,such
such identified
as as
viral entry,
viral from natural products
replication,
entry, replication, assembly,
assembly,have
and shed
and lightason
release,
release, well
as
how
well they
as interact
targeting with
of the viral life
virus–host‑specific cycle, such as viral
interactions entry,
[5,6]. replication,
The antiviral
as targeting of virus–host-specific interactions [5,6]. The antiviral properties of PTs have attracted the assembly,
properties andof release,
PTs as
have
well as targeting of virus–host‑specific
attracted
attention of the
many attention of manyPompei
researchers. andinteractions
researchers. Pompei and[5,6].
colleagues The antiviral
colleagues
extracted extractedproperties
glycyrrhizic glycyrrhizic ofacid
acid (Figure PTs(figure
have
2) (7) from
attracted the attention of many researchers. Pompei and colleagues extracted glycyrrhizic acid (figure
the2) (7) from
crude the crude
extract extract of
of Glycyrrha Glycyrrha
glabra roots,glabra
and roots,
it wasand it was
found to found to beagainst
be active active against the herpes
the herpes simplex
2) (7) fromvirus
simplex the crude extract of Glycyrrha glabra roots, and it was found to be active against the herpes
virus [15]. Chen et[15]. Chenisolated
al., also et al., analso isolated
oleanane-type an oleanane-type triterpene
triterpene derivative known derivative known
as salaspermic as
acid
simplex
salaspermicvirus acid[15]. Chen
(Figure 2) et
(8) al.,
from also isolated
Tripterygium an oleanane-type
wilfordii, which triterpene
blocked the derivative
replication of known
HIV in as
H9
(Figure 2) (8) from Tripterygium wilfordii, which blocked the replication of HIV in H9 lymphocytes (IC50 :
salaspermic
lymphocytesacid (IC(Figure 2) (8) from Tripterygium wilfordii, which blocked the replication of HIV in H9
50: 10 μM) [16]. Toshihiro et al. also isolated betulinic acid (BA) (Figure 2) (9) from
10 µM) [16]. Toshihiro
lymphocytes et al. also isolated betulinic acid (BA) (Figure 2) (9) from the bark and leaves of
the bark and (IC 50: 10 μM) [16]. Toshihiro et al. also isolated betulinic acid (BA) (Figure 2) (9) from
leaves of Syzigium claviflorum, which showed a significant effect against HIV (EC50: 1.4
Syzigium
the claviflorum, which showed a significant effect against HIV (EC 50 : 1.4 µM) HIV[17].(EC
Since then,
μM)bark
[17].andSinceleaves
then,ofmanySyzigium claviflorum,
natural PTs havewhich showed
been reported atosignificant effect
have antiviral against
activities. 50: 1.4
many natural PTs have been reported to have antiviral activities.
μM) [17]. Since then, many natural PTs have been reported to have antiviral activities.
X
X

O
O H
HOOC H
HOHOOC O
HO
HO O O
HOOC
HO OO O
HO
HOOC O
HO
HO
O 7
HO OH 7
OH

X
X
H
H H H H
H X
H H H
O H X
H
HO O
HO H
HO 8 H
8 HO 9
H
9

X-COOH
X-COOH
Figure 2. Isolated antiviral compounds from plants.
Figure2.2.Isolated
Figure Isolated antiviral compoundsfrom
antiviral compounds fromplants.
plants.
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One
One natural
naturalPTPTis Oleanolic acidacid
is Oleanolic (OA)(OA)
(Figure 3), which
(Figure 3), possesses many interesting
which possesses biological
many interesting
activities, such
biological as antiviral
activities, such as[18], anti-inflammatory,
antiviral analgesic [18,19],
[18], anti-inflammatory, antibacterial
analgesic [20,21], anti-cancer
[18,19], antibacterial [20,21],
anti-cancer [22–26], anti-oxidation [27,28], antimicrobial [29–31], and cardioprotective activitieset[32].
[22–26], anti-oxidation [27,28], antimicrobial [29–31], and cardioprotective activities [32]. Chen al.
reported
Chen thatreported
et al. OA provides extraordinary
that OA protection against
provides extraordinary acute and
protection chronic
against liver
acute and injury, andliver
chronic can
be usedand
injury, as ancanoral medication
be used for the
as an oral treatment
medication forofthe
human liver disorders
treatment of human[33].
liverOA is isolated
disorders [33].from
OA
more than 1600 different plant species [33–37], and is non-toxic and moderately
is isolated from more than 1600 different plant species [33–37], and is non-toxic and moderately water-soluble [19].
Table 1 below[19].
water-soluble shows
Tablesome medicinal
1 below plants
shows some containing
medicinal OAcontaining
plants as an active
OA as constituent, and their
an active constituent,
biological activities. In this study we review modifications of the basic oleanolic acid structure
and their biological activities. In this study we review modifications of the basic oleanolic acid structure that
have been made.
that have been made.
30 29

19 20 21

12 18 22
17
11 OH
25 26 13
1 14 16 28
9
2 O
10 8 15

3
7
5 27
HO 4 10
6

24 23

Figure 3. Structure of oleanolic acid (OA).

Table 1. Several plants where OA was reported, the plant parts used, and their biological activities.
Table 1. Several plants where OA was reported, the plant parts used, and their biological activities.
Plant Species (Family) Biological Activity Plant Parts Used References
Plant Species (Family) Biological Activity Plant Parts Used References
Oleaeuropaea L. (Oleaceae) Anticancer, antimicrobial, anti-diabetic Fruits and leaves [22,28,37,38]
Oleaeuropaea L. (Oleaceae) Anticancer, antimicrobial, anti-diabetic Fruits and leaves [22,28,37,38]
Fabiana
Fabiana imbricataR.
imbricata R.etet P.
P.
Antiviral, antitumor, and antihyperlipidemic
(Solanaceae) Antiviral, antitumor, and antihyperlipidemic Leaves and flowers
Leaves and flowers [39]
[39]
(Solanaceae)
Syzygium
Syzygium aromaticum
aromaticum Antinociceptive,
Antinociceptive,Anti-inflammatory,
Anti-inflammatory, Flower buds and
Flower buds and leaves [18,39,40]
(Myrtaceae) antihypertensive, and antioxidant [18,39,40]
(Myrtaceae) antihypertensive, and antioxidant leaves
Ligustrum lucidum Ait Anti-inflammatory, antioxidative,
Ligustrum lucidum Ait Anti-inflammatory, antioxidative, antiprotozoal, Fruits and leaves [41]
(Oleaceae) antiprotozoal, antimutagenic, and anticancer Fruits and leaves [41]
(Oleaceae) antimutagenic, and anticancer
Viscum album (Santalaceae) Anti-tumor, analgesic, and anti-inflammatory Leaves and stems [19,41,42]
Viscum album (Santalaceae) Anti-tumor, analgesic, and anti-inflammatory Leaves and stems [19,41,42]
Phyllanthusamarus
Phyllanthus amarus
Anti-diabetes Leaves or aerial [43]
(Phyllanthaceae) Anti-diabetes Leaves or aerial [43]
(Phyllanthaceae)
Punica granatum
Punica L. (Punicaceae)
granatum L. Antioxidant activity Fruit [27] [27]
Antioxidant activity Fruit
(Punicaceae)
Rosmarinus officinalis L. Anti-inflammatory, hepatoprotective, Leaves, flowers,
[28]
Rosmarinus (Lamiaceae)
officinalis L. gastroprotective, antiulcer
Anti-inflammatory, hepatoprotective, stems, branches.
Leaves, flowers,
[28]
Gentiana lutea (Gentianaceae)
(Lamiaceae) Antimicrobial antiulcer
gastroprotective, Dried root and rhizome
stems, branches. [30]
Gentiana lutea Anti-inflammatory, antioxidative, Dried root and
L. camara (Verbenaceae) Antimicrobial
antiprotozoal
Leaves and flowers [41] [30]
(Gentianaceae) rhizome
L. camara chingii
Viburnum (Verbenaceae)
(Asteraceae) Anti-inflammatory, antioxidative, antiprotozoal
Antimicrobial Leaves and flowers
Leaves [44] [41]
Viburnum chingii
Siphonodon celastrineus Antimicrobial
Anti-inflammatory Leaves
Root bark, stem [45,46][44]
(Asteraceae)
(Celastraceae)
Siphonodon
Rosa laevigata celastrineus
(Rosaceae) Anti-inflammatory Leaves [47]
Anti-inflammatory Root bark, stem [45,46]
(Celastraceae)
Fructus Ligustri Lucidi (FLL) Anti-hepatitis Leaves [18]
Rosa laevigata (Rosaceae) Anti-inflammatory Leaves [47]
Fructus Ligustri Lucidi
Anti-hepatitis Leaves [18]
Analogues (FLL) of Oleanolic Acid
OA (10) has three active sites (i.e., the hydroxyl C-3 in ring A, the alkene C12-C13 in ring C and
1.1. Analogues of Oleanolic Acid
carboxylic acid C-28), which can be modified in order to change its physical structure and improve
OA (10) effects
its biological has three activeMany
[47–49]. sites (i.e., the hydroxyl
analogues C-3 inbeen
of OA have ringsynthesized
A, the alkeneandC12-C13 in ring
tested for C and
numerous
biological
carboxylic activities [50].
acid C-28), It has
which been
can reported in literature
be modified that OA
order to change itsisphysical
a good precursor molecule
structure and for
improve
its biological effects [47–49]. Many analogues of OA have been synthesized and tested for numerous
semi-synthetic modifications due to its multiple biological properties, availability, and low production
biological
cost activities [50]. Itethas
[51]. Nkeh-Chungag al.been reported
reported in literature and
the methylation that acetylation
OA is a good of precursor molecule
OA originally for
isolated
semi-synthetic modifications due to its multiple biological properties, availability, and low
Molecules 2018, 23, x 4 of 14
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production cost [51]. Nkeh-Chungag et al. reported the methylation and acetylation of OA originally
isolated from
production Syzygium
cost aromaticum (clove),
[51]. Nkeh-Chungag which resulted
et al. reported in two compounds
the methylation (Figureof4)OA
and acetylation (11)originally
and (12).
Both Syzygium
from compounds
isolated exhibited
aromaticum
from Syzygium better(clove),
(clove),
aromaticum in vivo/in
which whichvitro
resulted anti-inflammatory
in two
resulted compounds
in and(Figure
(Figure
two compounds membrane-stabilizing
4) (11)4)and
(11)(12).
andBoth
(12).
properties,
compounds respectively,
exhibited when
better in compared
vivo/in to
vitro OA [52].
anti-inflammatory and membrane-stabilizing
Both compounds exhibited better in vivo/in vitro anti-inflammatory and membrane-stabilizing properties,
respectively, when compared
properties, respectively, whentocompared
OA [52]. to OA [52].

OR
H
OR
O H O
H O
O
O H
H
O
H 11 R = -CH3
11 R = -CH3
12 R = -H
12 R = -H
Figure 4. 3-acetoxy, 28-methyloleanolic acid (11); 3-acetoxyoleanolic acid (12).
Figure 4.
Figure 3-acetoxy, 28-methyloleanolic
4. 3-acetoxy, 28-methyloleanolic acid
acid (11);
(11); 3-acetoxyoleanolic
3-acetoxyoleanolic acid
acid (12).
(12).
Modification of OA has resulted in compounds with biological activity, such as antidiabetic
Modification
activity. Yolanda et
Modification of OA
ofal.
OA has resulted
synthesized
has in compounds
analogues
resulted of OA
in compounds with biological
(three
with ethers activity,
and four
biological such assuch
esters
activity, antidiabetic
on hydroxyl activity.
C3 in
as antidiabetic
Yolanda
ring A, et
threeal. synthesized
esters from analogues
the carboxyl of OA
group (three
C-28,ethers
and and four
corresponding esters on hydroxyl
primary
activity. Yolanda et al. synthesized analogues of OA (three ethers and four esters on hydroxyl C3 C3
alcohol), in ring A,
derived three
from
in
esters
ring from
the reduction the carboxyl
of carboxylic
A, three esters group C-28, and
acid withgroup
from the carboxyl corresponding
LiAlHC-28,
4. Cinnamoyl primary alcohol),
ester (13) primary
and corresponding derived from
and ethylalcohol), the
ether (14) reduction
(Figure
derived 5)
from
of carboxylic
were found toacid
be with
the LiAlH
most .
PTP-1B
4 Cinnamoyl ester
inhibitors. (13)
The in and ethyl
vitro ether
inhibitory (14) (Figure
effect
the reduction of carboxylic acid with LiAlH4. Cinnamoyl ester (13) and ethyl ether (14) (Figure 5) of 5) were
compound found
13 to
was
be thefound
were mostand
significant, PTP-1B
to be inhibitors.
it substantially
the most PTP-1B The ininhibitors.
lowered vitro
bloodinhibitory
The ineffect
glucose vitro of
levels in compound 13 was
vivo experiments
inhibitory effect significant,
when
of compound 13and
compared it
to
was
substantially
OA. Compound
significant, lowered blood glucose
14 exhibitedlowered
and it substantially levels
better bloodin vivo
inhibitory experiments
activity
glucose levels in when
andvivo compared
selectivity
experiments to OA.
overwhen Compound
protein-tyrosine
compared to 14
exhibited
phosphatasebetter 1B inhibitory
(PTP-1B), activity
with and
advanced selectivity over
interaction protein-tyrosine
with site B, in
OA. Compound 14 exhibited better inhibitory activity and selectivity over protein-tyrosine phosphatase
accordance with 1B (PTP-1B),
docking with
studies
advanced
[53].
phosphatase interaction with site B, in accordance with docking studies [53].
1B (PTP-1B), with advanced interaction with site B, in accordance with docking studies
[53].

H
H OH
OH
O
H
O 13 R= O
RO H
H 13 R= O
RO 14 R= C2H5
H
14 R= C2H5
Figure 5. (3b)-3-{[(2E)-3-phenylprop-2-enoyl]oxy}olean-12-en-28-oic
Figure 5. (3b)-3-{[(2E)-3-phenylprop-2-enoyl]oxy}olean-12-en-28-oic acid (13),
acid (13), (3b)-3-ethoxyolean-12-
(3b)-3-ethoxyolean-12-en-28-oic
en-28-oic acid (14). acid (14).
Figure 5. (3b)-3-{[(2E)-3-phenylprop-2-enoyl]oxy}olean-12-en-28-oic acid (13), (3b)-3-ethoxyolean-12-
en-28-oic acid (14).
The
The modification
modification of of oleanolic
oleanolic acid
acid also
also resulted
resulted in in potent
potent antibacterial
antibacterial agents. Hichri et
agents. Hichri et al.
al.
explored
explored the effect
effect of
of introducing an acyl
acyl substituent at
at the hydroxyl C-3
C-3 in
in ring
ring AA of
of OA. AA sequence
The themodification introducing
of oleanolican acid substituent
also resulted the hydroxyl
in potent antibacterial agents.OA.Hichri
sequence
et al.
of
of diverse
diverse triterpenic
triterpenic acid
acid esters
esters were
were prepared
prepared from
from oleanolic
oleanolic acid
acid using
using suitable
suitable cyclic
cyclic anhydrides,
anhydrides,
explored the effect of introducing an acyl substituent at the hydroxyl C-3 in ring A of OA. A sequence
acid
acid chlorides,
chlorides, and N,N-dimethyl-4-aminopyridine (DMAP)
and N,N-dimethyl-4-aminopyridine (DMAP) as as aa catalyst
catalyst (Figure 66 and
and Table 2) 2) [29].
[29].
of diverse triterpenic acid esters were prepared from oleanolic acid using (Figure
suitable cyclicTable
anhydrides,
OA and
OA and its acylated
its acylated analogues were screened for
analogues were screened for their their antimicrobial
antimicrobial activity
activity against
against five fungal
fiveTable plant
fungal2)plant
acid chlorides, and N,N-dimethyl-4-aminopyridine (DMAP) as a catalyst (Figure 6 and [29].
pathogens,
pathogens, and two Gram-positive
and two analogues
Gram-positive and two Gram-negative bacteria. Compound 15 with sulfur and
OA and its acylated were and two Gram-negative
screened bacteria.activity
for their antimicrobial Compound 15five
against with sulfurplant
fungal and
chlorine
chlorine atom(s),
atom(s), ((3b)-3-((thiophene-2-carbonyl)oxy)-olean-12-en-28-oic
((3b)-3-((thiophene-2-carbonyl)oxy)-olean-12-en-28-oic acid, was
acid, was found
found toto be
be an
an
pathogens, and two Gram-positive and two Gram-negative bacteria. Compound 15 with sulfur and
effective
effective antibacterial
antibacterial agent,
agent, and
and the
the most
most active
active antifungal
antifungal compound.
compound. It exhibited
It exhibited good
good activity
activity
chlorine atom(s), ((3b)-3-((thiophene-2-carbonyl)oxy)-olean-12-en-28-oic acid, was found to be an
against A.
against
effective
niger,
niger, P.
P.italicum,
A.antibacterial italicum, P. digitatum,
agent,P.and
digitatum, A.
the mostA.flavus,
flavus,and T.T.harzianum.
active and harzianum.
antifungal compound. It exhibited good activity
against A. niger, P. italicum, P. digitatum, A. flavus, and T. harzianum.
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H
H H
H H
H H H H
H
OH OH H H
OH
OH OH
OH OH OH
OH OH
OH
OH OH
OH
O
O
O O
O O
H H O O
O X /DMAP O
O O
O
H
H H
H H
X
X
X X
XX /DMAP
/DMAP
X/DMAP
/DMAP/DMAP
/DMAP H
H H
H O O
O
HO HO H
H H
HO HO
HO HO H
HO H RO RO
H
H H
H H RO
RO RO
RO RO H H H
H
HH H
Figure
Figure 6.
Figure
6.
FigureModification
6.
Modification
6. of
of OA.
Modification of
OA. OA.
Figure
Figure
Figure 6. 6. Modification
Modification
6. Modification
Modification of of
of OA.
of OA.
OA. OA.
Table 2. Synthesis of oleanolic acid.
Table
Table 2.
Table
2.
TableSynthesis
2.
Synthesis of
of oleanolic
2. Synthesis of
oleanolic
Synthesis acid.
of oleanolic
acid.
oleanolic acid.
acid.
Table
Table
Table 2. 2. Synthesis
2. Synthesis
Synthesis of of oleanolic
of oleanolic
oleanolic acid.acid.
acid.
Compound R X 1
Compound R X Yield (%) 111 Yield
1 (%)
Compound
Compound
Compound R RR X XX Yield
Yield (%)
Yield 1(%)
(%) 111
Compound
Compound
Compound R R R XX X Yield
Yield (%) 1111(%)
Yield
(%)
O
O
O O
O
OO O
15 15 X X RCl RCl 98 98
15 15
15
15 15
15
X
X
X X
X RCl RCl
RCl
RCl RCl
RCl 98 98
98
98 98
98
X-S
X-S X-S
X-S
X-S X-S
X-S
Cl
Cl Cl
Cl OO
O O
O
Cl Cl
Cl O O
16 16
16 16
16 RCl
RCl RCl
RCl
RCl 94
94 94
94 94
16 16
16 RCl RCl
RCl 94 94
94
Ph
Ph
Ph Ph
Ph
Ph
Ph Ph
Ph
Ph
Ph
Cl
Cl Cl O
O
Cl O O
O
Cl Cl
Cl O O
17 17
17 17
17 17 RCl RCl
RCl RCl
RCl 91 91
91 91 91
17 17
17 RCl
RCl RCl
RCl 91
91 91
91
O
O O
O
Cl O
Cl Cl
Cl O O
Cl
Cl Cl
Cl
18 18
18 18
18
18
RCl
RCl RCl
RCl
RCl 95
95 95
95 95
18 18
18 RCl RCl
RCl 95 95
95
Cl Cl
Cl
Cl Cl
Cl Cl
Cl O
O
O O
O
Cl
Cl Cl O
O O
19 19 Cl
Cl Cl
Cl Cl RCl RCl 94 94
19
19 19
19 19 RCl
RCl RCl 94
94 94 94
19 19 RCl RCl
RCl 94 94
94
Cl Cl
Cl
Cl Cl
Cl Cl
Cl O
O
O O
O O O
O
O O O
O OO
O OO
OO O
20 20 O
O
O OO
OO O
O
O O
O 92 92
20
20 20 20
20 20 92 92
92 92
92 92
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
O
O O
O
O
O O O
O O
O
O
O O
21
21
21 21 21
21 82
82 82
82 82
21 21 O
O O
O 82 82
82
O
O O
COOH
COOH
COOH
COOH
COOH
COOH O
O O
O
O O
O
O
O O
O O
O O
O
O
O
O O O
O O
22
22 22
22 O
O O
O 91
91 91
91
22 22
22 22 O
O O 91 91
91 91
COOH
COOH
COOH
COOH O O
COOH
COOH O
O O
O O
O
O O
O O O
O
O O O OO O
OO O
23 23 O
O OO
O OO
O O O 85 85
23 23
23 23
23 23 85 85
85 85
85 85
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
O O O OO
O O
OO O
O O
O O
O OO
OO OO
O O
O
O
O O
O O
24
24 24
24 81
81 81
81
24 24
24 24 HOOC
HOOC 81 81
81 81
HOOC
HOOC
HOOC
HOOC Cl Cl
Cl Cl
Cl Cl
Cl
111Obtained percentage
11 Obtained yield of product.
1Obtained
1111 Obtained
11 Obtained
percentage
percentage yield
percentage yield
of of product.
product.
yield ofproduct.
product.
Obtained
1 Obtained
Obtained percentage
percentage
percentage yieldyield
of ofof product.
of product.
yieldyield
percentage product.
Molecules 2018, 23, 2300 6 of 14
Molecules
Molecules 2018,
2018, 23,
23, x
x 66 of
of 14
14

2. Phytochemical
2. Phytochemical Studies
Studies and Anti-Viral Activities
and Anti-Viral Activities of
of OA
of OA
OA

2.1.
2.1. Anti-HIV
Anti-HIV Activity
Activity
Due
Due to
to the increase in
the increase
increase the occurrence
in the
the occurrence of
occurrence of drug-resistant
of drug-resistant virus
drug-resistant virus strains,
strains, the improvement of
the improvement
improvement of effective
effective
treatments
treatments for the HIV infection is dependent on the identification of novel biomedical agents with
for the HIV infection is dependent on the identification of novel biomedical agents with
low
low toxicity.
toxicity. Synthesis
toxicity. Synthesis of
of oleanolic
oleanolic acid,
acid, as
as well
well as
as other
other closely-related
closely-related triterpenes,
triterpenes, such
such asas betulinic
betulinic
acid
acid and
and dihydrobetulinic
dihydrobetulinic acid,
acid, has
has led
led to
to anti-HIV
anti-HIV agents
agents [53–55].
[53–55]. Zhu
[53–55]. Zhu et et al. synthesized derivatives
al. synthesized derivatives
of
of OA. These authors modified the C12-C13 double bond of OA yielding compound 25, which
OA. These authors modified the C12-C13 double bond of OA yielding compound 25, which was
was
3-fold
3-fold more
more active than OA.
active than Esterification of
OA. Esterification of 25 with anhydrides
25 with anhydrides resulted
resulted inin compounds
compounds 26–28,
26–28, which
which
were
were 5-fold
5-fold more
more active
more active than
active than OA
than OA with
OA with 28 showing
28
with 28 showing remarkable
showing remarkable activity
remarkable activity [56]
activity [56](Figure
[56] (Figure7).
(Figure 7).
7).

X:
X:
25
25 H
H
H
H
H 26: HOOC
26: HOOC
H COOH
COOH O
O
27
27 HOOC
HOOC
X
X O
O O
O

25-28
25-28 28
28 HOOC
HOOC
O
O
Figure
Figure 7.
7. Oleanolic
Oleanolic derivatives
derivatives with
with anti-HIV
anti-HIV activity.
activity.

Compound
Compound 25 25 was
25 was further
further modified
modified by
by converting
converting the
the C28-carboxyl
C28-carboxyl group
group to
to an
an aminomethyl
aminomethyl
group, resulting in compounds 29 and 30, which were greater than 10-fold more active
group, resulting in compounds 29 and 30, which were greater than 10-fold more active
resulting in compounds 29 and 30, which were greater than 10-fold when
more compared
active when
when
to OA (Figure
compared
compared to 8).
to OA
OA (Figure
(Figure 8).
8).

Z
Z Y
Y X
X
H
H

X
X Y
Y O
O
29
29

Z
Z Y
Y X
X
H
H

X Y O X-
X- COOH
COOH
X Y O
30
30 Y- C=O
Y- C=O
Z-
Z- NH
NH
Figure 8.
Figure
Figure 8. Previously derived
8. Previously
Previously derived OA
derived OA analogues.
OA analogues.
analogues.

Yu
Yu et al. in their structure-activity relationship study of effective anti-HIV agents, synthesized and
Yu etet al.
al. in
in their
their structure-activity
structure-activity relationship
relationship study
study ofof effective
effective anti-HIV
anti-HIV agents,
agents, synthesized
synthesized
evaluated
and evaluatednew new
evaluated triterpene derivatives
triterpene in vitro
derivatives for antiviral
in vitro
vitro activity.
for antiviral
antiviral OA analogue
activity. OA compound 31 was
and new triterpene derivatives in for activity. OA analogue
analogue compound
compound 31 31
inactive,
was whilewhile
OA derivative 32 exhibited an EC50an value ofvalue
0.32 µM, indicating that OA isthat
a promising
was inactive, while OA derivative 32 exhibited an EC50 value of 0.32 μM, indicating that OA is a
inactive, OA derivative 32 exhibited EC 50 of 0.32 μM, indicating OA is a
anti-HIV
promising inhibitor [56,57]. These compounds are potential therapeutics that would benefit from further
promising anti-HIV
anti-HIV inhibitor
inhibitor [56,57].
[56,57]. These
These compounds
compounds are are potential
potential therapeutics
therapeutics that
that would
would benefit
benefit
studies in vivo
from further
further (Figure
studies in 9).
vivo (Figure
(Figure 9).
9).
from studies in vivo
Molecules 2018, 23, 2300 7 of 14
Molecules 2018, 23, x 7 of 14

X
O
H O
RO O 31 R= OH
H
O
O O
32 R= OH
X- COOH
Figure 9.
Figure Previously modified
9. Previously modified anti-HIV
anti-HIV triterpene
triterpene derivatives.
derivatives.

Kashiwada
Kashiwada et et al.
al. prepared
prepared several
several 3-O-acyl-ursolic
3-O-acyl-ursolic acids
acids and
and evaluated
evaluated their
their anti-HIV
anti-HIV activity.
activity.
The most potent compound indicated
The most potent compound indicated an EC50 an EC value of 0.31 µM and a TI of 155.5 [58]. In another
50 value of 0.31 µ M and a TI of 155.5 [58]. In another
report by Kashiwada et al., OA derivatives inhibited
report by Kashiwada et al., OA derivatives inhibited HIV-1 HIV-1 replication
replication in
in acutely
acutely infected
infected H9
H9 cells.
cells.
OA-triterpenes isolated from the leaves of S. claviflorum exhibited potent anti-HIV activity,
OA-triterpenes isolated from the leaves of S. claviflorum exhibited potent anti-HIV activity, which which
further
further revealed
revealed the
the potential
potential of of OA
OA derivatives
derivatives for
for the
the treatment
treatment of
of HIV
HIV [59,60].
[59,60].
2.2. Anti-Influenza
2.2. Anti-Influenza
The influenza virus is a lethal respiratory virus capable of triggering significant damage to the
The influenza virus is a lethal respiratory virus capable of triggering significant damage to the
population. Vaccines and antiviral agents are important for controlling the outbreak of influenza. The
population. Vaccines and antiviral agents are important for controlling the outbreak of influenza. The
research into plant-based drugs against the influenza virus is promising, as some plants have been
research into plant-based drugs against the influenza virus is promising, as some plants have been
proven to have anti-influenza properties, some of these include: Aster spathulifolius, Pinus thunbergia,
proven to have anti-influenza properties, some of these include: Aster spathulifolius, Pinus thunbergia,
Thuja orientalis [61], Allium fistulosum [62], Sambucus nigra [63], and Psidium guajava [64]. OA and other
Thuja orientalis [61], Allium fistulosum [62], Sambucus nigra [63], and Psidium guajava [64]. OA and other
molecules, such as chlorogenic acid baicalein and quercetin, are regarded as active constituents in
molecules, such as chlorogenic acid baicalein and quercetin, are regarded as active constituents in
traditional Chinese folk medicine, which have been proven to be effective antiviral agents according to
traditional Chinese folk medicine, which have been proven to be effective antiviral agents according
clinical data [65]. These molecules have been found to be potential neuraminidase inhibitors, which
to clinical data [65]. These molecules have been found to be potential neuraminidase inhibitors, which
could be helpful for anti-influenza medicine development [66]. Yang et al. also reported that OA works
could be helpful for anti-influenza medicine development [66]. Yang et al. also reported that OA
as a broad-spectrum entry inhibitor of influenza viruses [67]. Han et al. conjugated sialic acid with
works as a broad-spectrum entry inhibitor of influenza viruses [67]. Han et al. conjugated sialic acid
OA and its analogues (Figure 9). In vitro evaluation of the compounds on the influenza A/WSN/33
with OA and its analogues (Figure 9). In vitro evaluation of the compounds on the influenza A/WSN/33
(H1N1) virus in MDCK cell culture revealed compound 33 as the most potent compound, with an IC50
(H1N1) virus in MDCK cell culture revealed compound 33 as the most potent compound, with an IC50 of
of 41.2 µM. It acted as an influenza virus entry inhibitor by inhibiting the binding of the influenza
41.2 μM. It acted as an influenza virus entry inhibitor by inhibiting the binding of the influenza virus
virus hemagglutinin protein to host cells [68]. In a similar report by Han et al., compounds 34 and
hemagglutinin protein to host cells [68]. In a similar report by Han et al., compounds 34 and 35 displayed
35 displayed anti-influenza activity against the A/WSN/33 (H1N1) virus. C-5 acetylamide and C-9
anti-influenza activity against the A/WSN/33 (H1N1) virus. C-5 acetylamide and C-9 hydroxy of
hydroxy of sialic acid were useful for their binding with hemagglutinin during viral entry into host
sialic acid were useful for their binding with hemagglutinin during viral entry into host cells [69]
cells [69] (Figure 10).
(FigureMolecules
10). 2018, 23, x 8 of 14

X
X
X
H
N
O
O OH O
O
O X-OH
HO 33

Figure 10. Cont.

X

H X OMe
N O X COOMe
N O
O N N NH
X
 O OH O
O
O X-OH
HO
Molecules 2018, 23, 2300 33 8 of 14

H X OMe
N O X COOMe
N O
O N N NH
X
HO
X-OAc
34

H X OMe
N O X COOMe
N O
O N N NH
X
HO
X-OAc
35
Figure
Figure 10.10.
OA OA derivatives
derivatives with
with anti-influenza
anti-influenza activity.
activity.

2.3. Anti-Hepatitis
2.3. Anti-Hepatitis
Previous reports estimated that approximately 170 million people across the world are infected
Previous reports estimated that approximately 170 million people across the world are infected
with the hepatitis C virus (HCV), which causes deaths globally [70]. Since the formation of Ribavirin
with the hepatitis C virus (HCV), which causes deaths globally [70]. Since the formation of Ribavirin
(RBV) and PEGylated interferon alpha (IFN-α) as the standard drugs for the medical treatment of
(RBV) and PEGylated interferon alpha (IFN-α) as the standard drugs for the medical treatment of
HCV in 1990, large efforts have been made to find new and more effective drugs. New effective
HCV in 1990, large efforts have been made to find new and more effective drugs. New effective HCV
HCV inhibitors belonging to direct-acting antivirals (DAAs), which include Boceprevir, Ledipasvir,
inhibitors belonging to direct-acting antivirals (DAAs), which include Boceprevir, Ledipasvir,
Sofosbuvir, Telaprevir, Simeprevir, and Daclatasvir, have been developed in recent years [71]. Although
Sofosbuvir, Telaprevir, Simeprevir, and Daclatasvir, have been developed in recent years [71].
great progress in anti-HCV remedies has been surely attained, many barriers nevertheless exist.
Although great progress in anti-HCV remedies has been surely attained, many barriers nevertheless
Monotherapy with DAAs is linked with the rapid occurrence of drug-resistant viral mutations, thus
exist. Monotherapy with DAAs is linked with the rapid occurrence of drug-resistant viral mutations,
the medical utility of DAAs is usually restricted to mixture regimens, which is associated with extra
thus the medical utility of DAAs is usually restricted to mixture regimens, which is associated with
side effects, drug-drug interactions, high costs, and availability. Therefore, new therapeutic techniques
extra side effects, drug-drug interactions, high costs, and availability. Therefore, new therapeutic
consisting of various HCV inhibitors focused on particular stages of the HCV life cycle, with more
techniques consisting of various HCV inhibitors focused on particular stages of the HCV life cycle,
effectiveness and wider availability, are still in demand to overcome those obstacles. Historically, many
with more effectiveness and wider availability, are still in demand to overcome those obstacles.
active modern drugs have been established from compounds initially isolated from plants, and now
Historically, many active modern drugs have been established from compounds initially isolated
there is still more interest in finding new drugs from herbal sources for the treatment of various human
from plants, and now there is still more interest in finding new drugs from herbal sources for the
diseases. Many natural compounds have been identified with antiviral effects globally, which includes
treatment of various human diseases. Many natural compounds have been identified with antiviral
anti-HCV activity. OA was an important compound in many traditional Chinese hepatic protective
effects globally, which includes anti-HCV activity. OA was an important compound in many
drugs, and was considered to play a therapeutic role in many diseases. Indeed, Plantago major L.,
traditional Chinese hepatic protective drugs, and was considered to play a therapeutic role in many
a well-known traditional Chinese remedy, has been used for the treatment of many diseases including
diseases. Indeed, Plantago major L., a well-known traditional Chinese remedy, has been used for the
the cold and viral hepatitis [72]. OA and its isomer, ursolic acid, were identified as active compounds of
treatment of many diseases including the cold and viral hepatitis [72]. OA and its isomer, ursolic acid,
Fructus Ligustri Lucidi (FLL), and were indicated as two antiviral compounds that seriously suppressed
were identified as active compounds of Fructus Ligustri Lucidi (FLL), and were indicated as two
the duplication of the Hepatitis C Virus (HCV) genotype 1b replicon, and the HCV genotype 2a JFH1
virus. Furthermore, these compounds exhibited anti-HCV properties, partly by suppressing HCV
NS5B RdRp properties, as noncompetitive inhibitors. Therefore, their results confirmed that natural
products of OA and ursolic acid are potential HCV antivirals [18].
Yan et al. (2006) reported interesting work on the synthesis of OA structures (Figure 11). These
researchers proposed a series of OA analogues which were synthesized by various reactions. All tested
analogues (38–41) inhibited the secretion of HBsAg, and also decreased the secretion of HBeAg [73].
Compound 39 showed major inhibition of HBV DNA duplication, which was significant when
compared to the reference drug. Due to its great performance in vivo and in vitro, compound 39
noncompetitive inhibitors. Therefore, their results confirmed that natural products of OA and ursolic
acid are potential HCV antivirals [18].
Yan et al. (2006) reported interesting work on the synthesis of OA structures (Figure 11). These
researchers proposed a series of OA analogues which were synthesized by various reactions. All
tested analogues
Molecules (38–41) inhibited the secretion of HBsAg, and also decreased the secretion of HBeAg
2018, 23, 2300 9 of 14
[73]. Compound 39 showed major inhibition of HBV DNA duplication, which was significant when
compared to the reference drug. Due to its great performance in vivo and in vitro, compound 39 is a
is a potential
potential novelnovel anti-Hepatitis
anti-Hepatitis B virus
B virus drug
drug candidate,
candidate, witha adifferent
with differentmechanism
mechanismof
of action,
action, which
which
needs further investigation.
needs further investigation.

O
39
O
H

(iv)

R R O
R
O (i) O (iii)
H O
R
R R 36 R 37
H OA H H

(iii)
(ii) (v)

R O
R R
O
O O
H O
R
O R
H 41 H 38 O
H 40

R= -OH

Figure 11. Synthesis route to derivatives.

Figure 11. Reagents and conditions: (i) N-Bromosuccinimide
derivatives. Reagents N-Bromosuccinimide (NBS), (NBS),
CCl , light, reflux, 4 h; (ii) chromic acid solution, acetone, 0 ◦ C, 1 h; (iii) Eosin Y, dichloromethane, light,
CCl4, light, reflux, 4 h; (ii) chromic acid solution, acetone, 0 °C, 1 h; (iii) Eosin Y, dichloromethane,
10 h; (iv)
light, chromic
10 h; acid solution,
(iv) chromic acetone,
acid solution, 0 ◦ C, 10h;
acetone, °C,(v)1 chromic acid solution,
h; (v) chromic acetone,
acid solution, 0 ◦ C, 10h.
acetone, °C, 1 h.

2.4. Anti-Herpes
2.4. Anti-Herpes
Herpes simplex virus (HSV) has two serotypes (HSV-1 and HSV-2), which target the oral and
Herpes simplex virus (HSV) has two serotypes (HSV-1 and HSV-2), which target the oral and
genital mucous membranes of humans, create latent infections in the sensory neurons, and may
genital mucous membranes of humans, create latent infections in the sensory neurons, and may
reactivate to cause recurring infections at the primary site [74]. The effects of genital herpes as a human
reactivate to cause recurring infections at the primary site [74]. The effects of genital herpes as a
health threat are increasing because of its interrelationship with HIV. HSV has been well treated with
human health threat are increasing because of its interrelationship with HIV. HSV has been well
acyclovir since 1970 [75]. Some licensed anti-herpes virus drugs, such as ganciclovir, cidofovir, and
treated with acyclovir since 1970 [75]. Some licensed anti-herpes virus drugs, such as ganciclovir,
foscarnet, that destroy herpes virus DNA polymerases, also have toxicity in long-term usage [76].
cidofovir, and foscarnet, that destroy herpes virus DNA polymerases, also have toxicity in long-term
Therefore, novel antiviral drugs from natural sources that have different mechanisms of action are in
great demand. Mukherjee et al. isolated OA from methanol (MeOH) extract taken from Achyranthes
aspera roots. The MeOH extract exhibited a weak anti-herpes virus effect (EC50 64.4 g/mL for Herpes
simplex virus 1 (HSV-1) and 72.8 g/mL for Herpes simplex virus 2 (HSV-2)), whereas OA showed
potent anti-herpes virus activity against both HSV-1 (EC50 6.8 g/mL) and HSV-2 (EC50 7.8 g/mL) [74].
Keda et al., also examined 15 oleanane-type triterpenoids, and their derivatives, for anti-herpes simplex
virus type 1 (HSV-1) activities. OA and its derivative Hederagenin (Figure 12) (42) were found to
exhibit moderate anti-HSV-1 activity [77].
Achyranthes aspera roots. The MeOH extract exhibited a weak anti-herpes virus effect (EC50 64.4 g/mL
for Herpes simplex virus 1 (HSV-1) and 72.8 g/mL for Herpes simplex virus 2 (HSV-2)), whereas OA
showed potent anti-herpes virus activity against both HSV-1 (EC50 6.8 g/mL) and HSV-2 (EC50 7.8
g/mL) [74]. Keda et al., also examined 15 oleanane-type triterpenoids, and their derivatives, for anti-
herpes simplex
Molecules virus type 1 (HSV-1) activities. OA and its derivative Hederagenin (Figure 12)
2018, 23, 2300 (42)
10 of 14
were found to exhibit moderate anti-HSV-1 activity [77].

OH
H
O
H
HO
H
42
OH

Figure 12. Hederagenin.

3. Conclusions
3. Conclusions
Triterpenoids
Triterpenoidsare areananimportant
importantgroup
groupofofcompounds
compounds widely
widely found
found in in
nature. Previous
nature. Previous reports on
reports
triterpenoids
on triterpenoids showed
showed that OA
that OAandanditsits
analogues
analogueshavehavecountless
countlessbeneficial
beneficialeffects,
effects, such
such as antiviral,
as antiviral,
anti-inflammatory,
anti-inflammatory, antitumor promotion, anticancer, and so forth. OA is relatively non-toxic and
antitumor promotion, anticancer, and so forth. OA is relatively non-toxic and has
has
been sold in
been sold in China
Chinaas asaatherapeutic
therapeuticfor forthethetreatment
treatment ofof human
human hepatitis.
hepatitis. There
There areare
fewfew reports
reports on
on the
the antiviral activities of OA and its analogues. The treatment of viral diseases
antiviral activities of OA and its analogues. The treatment of viral diseases continues to present continues to present
problems
problems in incurrent
currentmedicine,
medicine,with
withvarious
various studies showing
studies showing a great increase
a great in the
increase inoccurrence of drug
the occurrence of
resistant viruses [78]. The modification of the hydroxyl and carboxylic acid functional
drug resistant viruses [78]. The modification of the hydroxyl and carboxylic acid functional groups groups on OA
has
on OAresulted in potentin
has resulted compounds with antiviral
potent compounds withactivity.
antiviralThe anti-HIV
activity. Theactivity of the
anti-HIV synthesized
activity of the
compounds
synthesized compounds is via the inhibition of HIV-1 replication. The compounds synthesizedvirus
is via the inhibition of HIV-1 replication. The compounds synthesized also hindered also
entry by inhibiting the binding of influenza virus hemagglutinin protein to the host
hindered virus entry by inhibiting the binding of influenza virus hemagglutinin protein to the host cells. cells. However,
aHowever,
thorough antiviral antiviral
a thorough mode ofmodeactionofofaction
these of
compounds needs further
these compounds investigation.
needs further The initial
investigation. The
information obtained
initial information from infrom
obtained vivo and
in vivo in vitro
and studies
in vitroofstudies
OA derivatives is promising.
of OA derivatives Therefore,
is promising.
further studies
Therefore, are needed
further studiesinto
are OA, and its
needed intoanalogues
OA, andfrom natural products,
its analogues as an antiviral
from natural products,agent
as for
an
various human diseases.
antiviral agent for various human diseases.
Contributions: V.K.,
Author Contributions: V, Khwaza,
B.A.A. Blessing Aderibigbe
and O.O.O. and O.
contributed Oyedeji contributed
substantially substantially
to the work to the
reported. The work
authors
approved the final
reported. The version
authors of the the
approved manuscript.
final version of the manuscript.

Funding: This
Funding: research
This waswas
research funded by South
funded African
by South Medical
African Research
Medical CouncilCouncil
Research (Self-Initiated Research),
(Self-Initiated National
Research),
Research Foundation South African and Govan Mbeki Research and Development Centre (GMRDC), University
National Research Foundation South African and Govan Mbeki Research and Development Centre (GMRDC),
of Fort Hare.
University of Fort Hare.
Conflicts of Interest: The authors declare no conflict of interest regarding the publication of this paper.
Acknowledgments: The authors acknowledge Govan Mbeki Research and Development Centre (GMRDC),
University of Fort Hare, and NRF–Sasol Inzalo for financial support. Medical Research Council (Self-Initiated
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