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Recent advances in anti-HIV natural products

Inder Pal Singh* and Hardik S. Bodiwala
Received 12th August 2010
DOI: 10.1039/c0np00025f

Covering: 2004 to April 2010

This review covers the recent advances in anti-HIV natural products of various origins and chemical
classes. The literature of the last seven years has been thoroughly reviewed, with around 160
Published on 25 October 2010 on http://pubs.rsc.org | doi:10.1039/C0NP00025F

compounds discussed and 141 references cited. Some molecules in an advanced stage of development,
such as calanolide A and bevirimat, are also mentioned.

1 Introduction due to AIDS and preventing new infections. The number of new
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2 Natural products from plants HIV infections declined from 3.0 million in 2001 to 2.7 million in
2.1 Alkaloids 2008,1 and the efforts of the World Bank, WHO and G8 decla-
2.1.1 Newly discovered anti-HIV alkaloids ration for universal access to treatment can be regarded as the
2.1.2 Known alkaloids discovered to be anti-HIV agents key factors for this positive result.2 However, overall, 2.0 million
2.2 Phenolics people died in 2008 due to HIV infection compared with an
2.2.1 Newly discovered anti-HIV phenolics estimated 1.7 million in 2001,1 a fact that can be attributed to the
2.2.2 Known phenolics discovered to be anti-HIV agents higher number of infections in the early part of this decade
2.3 Terpenes coupled with lesser availability of newer and efficacious anti-
2.3.1 Newly discovered anti-HIV terpenes retroviral drugs during those years in the sub-Saharan region.
2.3.2 Known terpenes discovered to be anti-HIV agents Currently, the treatment available for the disease is known as
2.4 Proteins and polysaccharides HAART (Highly Active Anti-Retroviral Therapy). However,
2.5 Miscellaneous compounds due to complexity and social stigma associated with HIV in
3 Natural products of microbial origin developing countries, the long-term management of HIV
3.1 Newly discovered anti-HIV agents of microbial patients is difficult. The potent drugs may fail at a later stage
origin because of chronic adverse effects and the emergence and
3.2 Known microbial metabolites discovered to be anti- transmission of drug-resistant variants. This underlines the need
HIV agents for continuous efforts in the discovery of new antiretroviral/anti-
4 Natural products of marine origin HIV agents.3
5 Advanced-stage molecules – where are they now? Many of the reviews published earlier on this topic are mainly
6 Conclusions and future perspectives on a specific class of compounds, or natural products active
7 Acknowledgement against specific targets, such as integrase. For example, there are
8 References reviews with limited coverage such as those on coumarins,4 tri-
terpenes,5 flavanoids,6 integrase inhibitors at Merck,7 and
natural products with anti-HIV activity from marine sponges.8
The most recent review by Cos et al. covers natural anti-HIV
1 Introduction compounds reported during 1997–2007,9 although many of the
references are either prior to 2005 or synthetic modifications or
Human Immunodeficiency Virus (HIV) / AIDS remains a major
further mechanistic studies on previously isolated anti-HIV
health problem worldwide that has already caused an estimated
compounds. The present review provides extensive coverage of
25 million deaths and has generated profound demographic
all natural products obtained over the last seven years (2004–
changes in the most heavily affected countries in the last 27 years.
2010) from diverse sources that exhibit anti-HIV activity towards
Globally, there were an estimated 33 million people living with
different biological targets.
HIV in 2008. Young people between 15 to 24 years of age
The HIV replication cycle is summarized in Fig. 1. The first
account for an estimated 45% of the new HIV infections
step in the life-cycle involves virus fusion to the host cell through
worldwide. A 6-fold increase in financing for HIV programmes
viral glycoprotein gp120 and CD4 receptors. Then, gp120
in low- and middle-income countries is beginning to give results,
interacts with the chemokine co-receptor (either CCR5 or
with many countries making major progress in lowering deaths
CXCR4). This is followed by release of the viral RNA into the
host cell cytoplasm. By action of the reverse transcriptase
Department of Natural Products, National Institute of Pharmaceutical enzyme, viral RNA is transcribed into double-stranded viral
Education and Research (NIPER), Sector-67, S.A.S. Nagar, Punjab,
160062, India. E-mail: ipsingh@niper.ac.in; ipsingh67@yahoo.com; Fax: DNA that migrates into the host cell nucleus. This viral DNA is
+91 172-2214692; Tel: +91 172-2292144 then incorporated into the host cell chromosome by the action of

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Fig. 2 Newly discovered anti-HIV alkaloids.

Fig. 1 The HIV life-cycle showing the different targets.
integrase. Following this, various viral proteins and viral
genomic material are produced after transcription and trans-
lation by the host cell. Finally, two copies of viral RNA and
other proteins are incorporated into the immature virus, which is
then released and converted into a mature and functionally active
virus by the Gag polyprotein.
2 Natural products from plants
2.1 Alkaloids
Alkaloids of various structural subclasses have been reported to
possess anti-HIV activity. During the coverage period, few new
alkaloids were reported as anti-HIV agents, but many known
alkaloids were evaluated and discovered to be anti-HIV agents.
The structures of anti-HIV alkaloids are shown in Fig. 2 and 3.
2.1.1 Newly discovered anti-HIV alkaloids. A diprenylated
indole alkaloid, (E)-3-(3-hydroxymethyl-2-butenyl)-7-(3-methyl-
2-butenyl)-1H-indole (1), was isolated from the twigs and leaves
of Glycosmis montana native to China. The alkaloid 1 exhibited
potent anti-HIV activity with an IC50 value of 1.17 mg/mL and an
SI of 11.68. It was suggested that these diprenylated indole
alkaloids might be candidates for further study as potential anti-
HIV agents.10 A sesquiterpene-pyridine alkaloid, tripfordine A
(2), was isolated from an ethanolic extract of the roots of
poisonous liana Tripterygium wilfordii. The highly toxic roots are
known as ‘lei gong teng’ in Chinese folklore and used as an
insecticide. Compound 2 possesses an unusual structure con-
taining a sesquiterpene skeleton joined to a pyridine ring by

Inder Pal Singh obtained his Hardik S. Bodiwala was born

Ph.D. from Punjab Agricultural
University, Ludhiana, India. In
1994, he moved to Shizuoka
University, Japan, on a Mon-
busho Fellowship and obtained
his second Ph.D. under the
guidance of Prof. Hideo Etoh.
He worked as a post-doctoral
associate (1998–2000) in Prof.
Gerwick’s laboratory at Oregon
State University, USA. Later,
he joined Institute of Chemical
Inder Pal Singh Research, Kyoto University,
Japan, on a JSPS fellowship for
a two-year period. He is
currently an Associate Professor at NIPER. His research interests
include isolation of bioactive molecules from natural sources,
biomimetic synthesis of bioactive natural products, and standard-
ization of herbal/Ayurvedic formulations.
in 1982 in Ahmedabad,
Gujarat, India. After
completing a B.Pharm. in
2003, he obtained his
M.S.(Pharm.) from NIPER in
2005. During this period, he
worked on the isolation and
characterization of antipara-
sitic compounds. In 2005 and
2006 he worked in the R&D
unit of Cadila Pharmaceuticals
Ltd, Ahmedabad, on method
Hardik S: Bodiwala development and analysis of
biological samples. He is
currently pursuing a Ph.D.
under the supervision of Inder Pal Singh on the discovery of
natural product based anti-HIV agents.

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Fig. 3 Known alkaloids discovered to be anti-HIV agents.
a macrocycle. Compound 2, along with the other sesquiterpene-
pyridine alkaloids, were screened against HIV-1. Tripfordine A
showed moderate anti-HIV activity, with an EC50 of 13.4 mg/mL
and TI of 1.9, while the other alkaloids exhibited more potent
activity, with EC50 values of 2.54 to <0.1 mg/mL. The SAR
suggested that the C-5 acetyl group may be essential for the
activity, as compounds with a 5-acetyl group were more active
than those with 5-hydroxy, 5-benzyloxy and 5-furanoyl groups.
The position of the carboxypropyl or carboxybutyl side chain
(C-20 or C-40 ) on the nicotinic moiety did not affect the activity
(EC50 <0.10 mg/mL). Further, the compounds with a tertiary
hydroxy group in the propionic (C-80 ) or butyric (C-90 ) acid side
chain showed anti-HIV activity, while those without an OH were
inactive or less active. However, one alkaloid hyponine B, which
lacks an OH on C-80 , was also highly active; hence, it was sug-
gested that other structural parameters may also play an
important role in the activity.11
Anibamine (3), a novel pyridine quaternary alkaloid, was
isolated from the CH3OH extract of the stems of an Aniba species
as a TFA salt. Anibamine competed for the binding of 125I-gp120
to human CCR5 with an IC50 of 1 mM.12 Molecular docking
studies of anibamine in two different CCR5 homology models
(bovine rhodopsin and human b2-adrenergic receptors)
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suggested that the binding site of anibamine had some features in
common with the other CCR5 antagonists, therefore suggesting
that 3 may bind in similar binding sites and/or modes.13
A new canthin-4-one type alkaloid, drymaritin (4), isolated from
Drymaria diandra (also known as Drymaria cordata subsp. diandra)
exhibited anti-HIV activity in H9 lymphocytes with an EC50 value
of 0.699 mg/mL and a TI of 20.6. The canthin-6-one type of alka-
loids have been already reported to have anti-HIV activity.14
2.1.2 Known alkaloids discovered to be anti-HIV agents. The
iboga-type indole alkaloid 18-methoxycoronaridine (18-MC) (5)
was semisynthesized from coronaridine, and inhibits retroviral
activity with EC50 of 22.5 and 23 mM for R5 and X4 isolates in
PBMC. Similarly, 5 inhibited HIV-1 replication in macrophages
with EC50 values of 12.8 and 9.5 mM and TI of 25.6 and 34.5 after
14 and 21 days of infection, respectively. It moderately inhibited
HIV-1 RT with an IC50 of 69.4 mM.15 The major metabolite of 5
in human liver microsomes was 18-hydroxycoronaridine
(18-HC). The metabolism of 18-MC to 18-HC was catalyzed by
the polymorphic enzyme CYP2C19, and studies of this suggested
that 18-MC may be a selective probe substrate of CYP2C19, one
that is perhaps more selective and specific than currently used
probes (omeprazole and mephenytoin).16 Two carbazole
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Fig. 4 Newly discovered anti-HIV phenolics.
alkaloids, glybomine B (6) and glycoborinine (7), were isolated
from the twigs and leaves of Glycosmis montana. Earlier these
alkaloids were reported from G. arborea.17,18 Both the alkaloids
exhibited weak to moderate in vitro inhibitory activity against
HIV replication in C8166 cells. Compounds 6 and 7 demon-
strated anti-HIV activity with an IC50 of 9.73 mg/mL and
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4.47 mg/mL, respectively.10 Other carbazole alkaloids, O-meth-
ylmukonal (8), 3-formyl-2,7-dimethoxycarbazole (9) and claus-
zoline J (10), were isolated from the rhizomes and roots of
Clausena excavata. Compounds 8–10 showed anti-HIV-1 activity
in a syncytial assay, with EC50 values of 12.0, 29.1 and 34.2 mM
and TI of 56.7, 8.0 and 1.6, respectively.19
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Fig. 5 Known phenolics discovered to be anti-HIV agents.

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Fig. 6 Known phenolics discovered to be anti-HIV agents (continued).

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The alkaloids decarine (11), g-fagarine (12) and (+)-temba-
mide (13) were the most potent anti-HIV compounds of 67
compounds isolated from the root bark of Zanthoxylum ailan-
thoides. Compounds 11–13 showed EC50 values of <0.1 mg/mL
and TI values of >226, >231, and >215, respectively. Fourteen
other compounds from the same plant showed significant activity
against HIV replication in H9 lymphocyte cells.20
Two benzylisoquinoline alkaloids, (+)-1-(R)-coclaurine (14)
and ()-1-(S)-norcoclaurine (15), isolated from the leaves of
Nelumbo nucifera, a perennial aquatic crop grown throughout
Asia, showed anti-HIV activity with EC50 values of 0.8 and
<0.8 mg/mL and TI values of >125 and >25, respectively.
Compound 15 is a precursor in the biosynthesis of well-known
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anti-HIV alkaloids such as berberine and aporphine, as well as
other biologically active benzylisoquinoline and benzophenan-
thridine alkaloids.21 Norcoclaurine synthase is the first enzyme in
the biosynthetic pathway to benzylisoquinoline alkaloids in
several plants.22 Some other bisbenzylisoquinoline alkaloids,
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liensinine (16), negferine (17) and isoliensinine (18), were evalu-
ated against HIV to establish the SAR. Compounds 16–18
showed anti-HIV activity with EC50 values of <0.8 mg/mL and TI
values of >9.9, >8.6 and >6.5, respectively, while the aporphine
alkaloid nuciferine (19) showed an EC50 of 0.8 mg/mL and TI of
36.23 Norruffscine (20) and ()-8-oxotetrahydropalmatine (21)
were isolated from Pericampylus glaucus, a climbing shrub widely
distributed in the southwest of China. Compounds 20 and 21
showed EC50 values of 10.9 and 14.1 mM and SI values of 45.7
and 18.8 respectively against HIV-1 in C8166 cells.24
Cycleanine (22) a bisbenzylisoquinoline alkaloid isolated from
the root bark of Epinetrum villosum, was active against HIV-2
(EC50 1.83 mg/mL); however, it was 10 times less active against
HIV-1.25 Cycleanine is also reported to show potent antibacte-
rial, antifungal, antiplasmodial, and cytotoxic activities. Tablets
containing 2 mg cycleanine caused inhibition of TPA-promoted
tumours in rat ear, as well as superoxide generation by poly-
morphonuclear leukocytes. Cycleanine-containing hair tonic was
shown to control dandruff and prevent alopecia.26 Amary-
llidaceae alkaloids lycorine (23) and homolycorine (24) isolated
from the bulbs of Leucojum vernum possess anti-HIV-1 activity in
MT4 cells with ID50 values of 0.4 and 7.3 mg/mL, respectively. In
addition, 23 was active in a mechanism-based anti-HIV-RT
assay, with an ID50 of 0.4 mg/mL.27 Lycorine is also reported to
show antimalarial activity towards Plasmodium falciparum.28 A
small molecule, indole-3-carboxylic acid (25), was among 44
known compounds isolated from rhizomes of Begonia nantoen-
sis, and was also active against HIV replication in H9 lympho-
cyte cells, with an EC50 of 2.41 mg/mL and a TI of 6.79.29
2.2 Phenolics
Various phenolics, such as calanolide A, DCK and DCQA, are
currently in clinical trials.30 The structures of anti-HIV phenolics
are shown in Figs. 4–6.
2.2.1 Newly discovered anti-HIV phenolics. An inseparable
2 : 1 mixture of new dibenzocyclooctadiene lignans rubrisandrins
A (26 + 27) was isolated from the fruits of Schisandra rubriflora,
indigenous to southern China. The two regioisomers were
distinguished by 1H, 13C and 2D-NMR data. The mixture
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showed in vitro inhibitory effects against HIV replication in H9
lymphocytes, with an EC50 of 11.3 mM.31 Longipedunin A (28)
was isolated from the roots and stems of Kadsura long-
ipedunculata, and showed an IC50 of 50 mg/mL against HIV-1
PR. The higher activity of 28 compared to other dibenzocy-
clooctadiene lignans indicated that a trans-cinnamic acid ester
group might be an important functional group for the anti-HIV
PR activity.32 Tiegusanin G (29) was one of 14 new lignans iso-
lated from a 70% aqueous acetone extract of the aerial parts of
Schisandra propinqua var. sinensis, popularly known as ‘tie-gu-
san’ in China. Compound 29 showed anti-HIV-1 activity in the
C8166 cell line, with an EC50 value of 7.9 mM and TI of > 25.33
New flavonoid glycosides 6-(g,g-dimethylallyl)dihydro-
kaempferol-7-O-b-D-glucoside (30), 6-(g,g-dimethylallyl)dihydro-
quercetin-7-O-b-D-glucoside (31), 6-(3-hydroxy-3-methylbutyl)
taxifolin-7-O-b-D-glucoside (32) and 6-(3-hydroxy-3-methylbutyl)-
quercetin-7-O-b-D-glucoside (33) were isolated from the anti-
HIV-active methanolic extract of Ochna integerrima. These
compounds showed anti-HIV-1 activity in the syncytium assay
with an EC50 of 29.1, 14.0, 102.4 and 45.8 mg/mL, respectively. In
the same study, vitexin (containing a sugar unit on ring A) was
found to be inactive. Earlier, Meragelman et al.34 had reported
that 6-isoprenylated flavonoids with two hydroxyls at C-5 and
C-7 were inactive while McKee et al.35 reported that compounds
with 30 -OH were active against HIV. This suggested that the
isoprenyl and sugar unit on ring A and 30 -OH (which are present
in the above compounds) are required for anti-HIV activity.36
Three phenolic compounds, 2,3-dihydroxy-4-methoxy-6,6,9-tri-
methyl-6H-dibenzo[b,d]pyran (34), 8-methoxy-2-methyl-2-(4-
methyl-3-pentenyl)-2H-1-benzopyran-6-ol (35) and 4-methoxy-
3-(3-ethyl-2-butenyl)benzoic acid (36) isolated from Wigandia
urens showed activity in the CCR5 assay with IC50 values of 33,
46 and 26 mM respectively. Compound 36 was previously
synthesized as an intermediate.37 Arzanol (37) is a phloroglucinol
a-pyrone derivative isolated form the acetone extract of Heli-
chrysum italicum ssp. microphyllum, one of the best-known
medicinal plants in the Mediterranean area. The structural
elucidation of arzanol led to a revision of the structure of
homoarenol. Arzanol inhibited TNFa-induced HIV-LTR
transactivation in a T-cell line with an IC50 of 5 mM, but failed to
inhibit luciferase activity in HeLa Tet-On-Luc assay.38
The dimeric lactone ardimerin digallate (38) isolated from the
whole plants of Ardisia japonica, showed inhibitory activity
against both HIV-1 and HIV-2 RNase H in vitro with IC50 values
of 1.5 and 1.1 mM, respectively. It did not inhibit human or
E. coli RNase H at 200 mM, demonstrating selectivity of 38
towards inhibition of the retroviral enzyme. A similar compound
lacking a galloyl moiety, ardimerin, did not show any inhibition
of RNase H, suggesting that the galloyl group is important for
anti-HIV activity.39
Three quinic acid derivatives, methyl-5-O-caffeoyl-3-O-sina-
poylquinate (39), methyl-5-O-caffeoyl-4-O-sinapoylquinate (41)
and methyl-3,5-di-O-caffeoyl-4-O-(3-hydroxy-3-methyl)glutar-
oylquinate (43), were isolated by bioactivity-guided fraction-
ation of the ethyl acetate extract of Gardenia jasminoides
(Gardeniae Fructus). Compounds 39, 41 and 43 showed HIV-1
IN inhibitory activity with an IC50 of 20 mg/mL. Ethyl quinate
derivatives 40 and 42 showed an IC50 of 45 mg/mL in the same
assay.40
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2.2.2 Known phenolics discovered to be anti-HIV agents.
Sixteen known compounds, including dibenzocyclooctadiene-
type lignans, were isolated from the stems of Kadsura heteroclita,
a plant native to southern China. Lignans kadsurin (44), heter-
oclitin F (45) and acetoxy oxokadsurane (46) were active as anti-
HIV agents. Compounds interiorin (47) and interiorin B (48)
exhibited anti-HIV activity with EC50 values of 1.6 and
1.4 mg/mL, and TI values of 52.9 and 65.9, respectively.
Comparison of 47 and 48 with other dibenzocyclooctadiene
lignans having similar skeletons indicated that the aa0 bb0 -dien-
one might be an important functional group for anti-HIV
activity.41
Dibenzocyclooctadiene lignans gomisin J (49), ()-gomisin
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M1 (50), (+)-gomisin M2 (51) and schisanhenol (52) were isolated
from the fruits of Schisandra rubriflora. ()-Gomisin M1 (50)
exhibited the most potent anti-HIV activity, with EC50 and TI
values of <0.65 mM and >68, respectively. It was found that
compounds with aromatic hydroxyl groups were more active
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than those lacking aromatic hydroxyls, suggesting the impor-
tance of the aromatic hydroxyl groups for the anti-HIV activity
of dibenzocyclooctadiene lignans. The position of the hydroxyl
was also important for enhancing the anti-HIV activity, as 50
(with a 30 -OH), was more potent than 51 (with a 3-OH).31 Four
known lignans, gomisin J (49), schizandrin (53), schisantherin A
(54) and schisantherin D (55), were isolated from the leaves and
stems of Schisandra sphenanthera. The anti-HIV-1 activity of 49
and 53–55 was evaluated by the inhibition assay for the cyto-
pathic effects of HIV-1IIIB. Compounds 49 and 53–55 showed
anti-HIV-1 activity, with EC50 values of 17.2, 15.5, 19.0 and
20.5 mg/mL, respectively.42 Gomisin J is also known to inhibit
L-type calcium channels and stimulate glucose uptake into
peripheral tissue.43,44 Anti-inflammatory activity in murine
macrophages has also been reported.45 Binakadsurin A (56) was
isolated from 70% aqueous acetone extract of stems of Kadsura
angustifolia, and was isolated along with 11 new lignans.
Compound 56 showed anti HIV-1 activity in the C8166 cell line,
with an EC50 of 3.86 mM, a CC50 of 227.16 mM and an SI of
58.92.46
Flavonoids quercetin (57), quercitrin (58) and myricetin 3-O-
b-D-galactopyranoside (59) were isolated from the EtOAc extract
of leaves of Alnus firma. The compounds showed inhibition
against HIV-1 RT with an IC50 of 60 mM, whereas they were
inactive against HIV-1 PR.47 5,7-Dihydroxychromone (60), and
()-catechin (61), isolated from Begonia nantoensis, endemic to
the mountains of central Taiwan, demonstrated activity against
HIV replication in H9 lymphocyte cells with EC50 values of 18.7
and 14.3 mg/mL, and TI values of 1.3 and 1.8, respectively.29
Eight prenylisoflavonoids were isolated from the DCM extract
of Erythrina senegalensis, a Cameroonian medicinal plant.
8-Prenylluteone (62), auriculatin (63), erysenegalensein O (64),
erysenegalensein D (65), erysenegalensein N (66), derrone (67),
alpinumisoflavone (68) and 6,8-diprenylgenistein (69) showed
HIV-1 PR inhibitory activity with IC50 values of 4.0, 3.5, 5.0, 2.5,
4.5, 18.2, 30.1 and 0.5 mM, respectively. The two prenyl groups at
the 6- and 8-positions, along with a hydroxy at the 40 -position,
were found to be important for the activity.48 Calycosin-7-O-b-D-
glucopyranoside (70) has been isolated from the roots of
Astragalus membranaceus var. mongholicus. This plant has been
used as traditional Chinese medicine to cure various diseases like
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diarrhoea, abnormal uterine bleeding, edema, anemia, albu-
minuria in chronic nephritis, and diabetes. Compound 70
inhibited cytopathic effects with an EC50 of 15.7 mM and TI of
28.5.49
Biflavonoids ochnaflavone 700 -O-methyl ether and 200 ,300 -
dihydroochnaflavone 700 -O-methyl ether, isolated from Ochna
integerrima, were found to be active in the syncytium assay with
EC50 values of 2.0 and 0.9 mg/ml, respectively. In a mechanism-
based assay, these compounds inhibited HIV-1 RT with IC50
values of 2.0 and 2.4 mg/mL, respectively.36
Orobol (71) and wedelolactone (72) isolated from Eclipta
prostrata showed activity against HIV-1 IN with IC50 values of
8.1 and 4.0 mM, respectively; but they did not inhibit HIV-1 PR.50
Wedelolactone possesses a wide range of biological activities,
including its use as an antidote for snake venom and hep-
atoprotective effects. It is also known to inhibit the IKK
complex.51 The cyclohexenyl chalcone hydroxypanduratin A (73)
has been obtained from the methanolic extract of rhizomes of
Boesenbergia pandurata, a perennial herb of the Zingiberaceae
family. This plant is locally known in Thailand as ‘kra-chai’, and
has been used as self-medication by AIDS patients in that
country. Compound 73 possesses anti-HIV-1 protease activity,
with an IC50 value of 5.6 mM. SAR of cyclohexenyl chalcones for
HIV-1 PR suggested that compounds with a hydroxyl group at
C-4 were more active than those with a methoxy group, preny-
lated compounds were more active than non-prenylated
compounds, hydroxylation at the 4000 -position reduced activity,
while introduction of a double bond at C10 and C60 increased the
activity against HIV-PR.52
Punicalin (74) and 2-O-galloylpunicalin (75), isolated from the
aqueous extract of Terminalia triflora leaves showed inhibitory
activity on HIV-1 RT, with an IC50 of 0.14 mM and 0.11 mM.
Both the tannins were previously isolated from the genus
Terminalia.53 Punicalins and punicalagins, as well as other
tannins, are well-known antioxidants and anticancer agents,54,55
and have also shown antimicrobial and antimalarial activities.56
Clausenidin (76), a pyranocoumarin, was isolated from the
rhizomes and roots of Clausena excavata and it displayed anti-
HIV-1 activity in a syncytial assay with an EC50 of 5.3 mM and
a TI of 7.0. Earlier, 76 was reported to be inactive in an HIV-RT
assay, so it might be acting on a different target in the HIV life-
cycle.19
4-Phenylcoumarins were isolated from lipophilic dichloro-
methane extract of leaves of Marila pluricostata collected from
Panama.57 Mesuol (77) affected neither the reverse transcription
nor integration steps of the viral cycle, however the antiviral
activity was additive when combined with AZT. A mechanistic
study revealed that mesuol inhibited TNFa-induced HIV-1–
LTR transcriptional activity by targeting the NF-kB pathway.
This hinted at the potential of the NF-kB transcription factor as
one of the targets for anti-HIV compounds.58 In addition,
compounds 78, 79 and 80 were found to be active in the NF-kB
and HeLa-Tat-Luc tests at 25 mM concentration. These
compounds were active against recombinant HIV with IC50
values of 6.9, 31.9 and 0.5 mM, respectively.59
The caged xanthones morellic acid (81), gambogic acid (82),
desoxygambogenin (83), forbesione (84) and dihydroisomorellin
(85) were isolated from the resin and fruits of Garcinia hanburyi.
Compounds 83 and 85 showed anti-HIV-1 activity in
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a syncytium assay with EC50 values of 3.0 and 1.2 mg/mL and TI
values of 1.7 and 4.7, respectively. Compounds 81, 82, 84 and 85
exhibited anti-HIV-1 RT activity, with IC50 values of 11.0, 15.0,
62.1 and 42.3 mg/mL.60 Two xanthones, 1,3,7-trihydroxy-6-
methoxy-4,5-di-(3-methylbut-2-enyl)xanthone (86) and euxan-
thone (87), were isolated form the MeOH extract of the aerial
part of Cratoxylum arborescens. Compounds 86 and 87 displayed
anti-HIV-1 RT activity with an IC50 of 84.9 and 50.1 mg/mL,
respectively.61
Polyisoprenylated benzophenones were isolated from the
fruits of Clusia torresii, a tree endemic to Costa Rica. Clusianone
(88), 7-epi-clusianone (89), 18,19-dihydroxyclusianone (90),
propolone A (91) and nemorosone (92) have shown anti-HIV
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activity with EC50 values of 0.02, 2.0, 7.1, 0.32 and 0.8 mM, and
SI values of 5.0, 10.0, 2.2, 15.6 and 6.2, respectively. Compounds
88 and 89 remain in constant equilibrium between their keto and
enol tautomers. The SAR suggested that presence of a keto–enol
equilibrium was not essential for anti-HIV activity, and that the
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C-7 configuration might play an important role in the activity. A
study of the mechanism of action revealed that benzophenone
derivatives of type B (88–90) inhibited the gp120-sCD4 interac-
tion, indicating that they might be interfering with the viral
attachment to the cellular receptor CD4, and thus preventing
infection. Compounds 91 and 92 had no effect on the gp120-
sCD4 interaction.62 Clusianone, first discovered in 1976, is also
known to have cancer-chemopreventive properties.63 Other
Fig. 7 Newly discovered anti-HIV terpenes.
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polyprenylated benzophenones structurally similar to clusianone
are reported to possess antioxidant activities.64
Long-chain phenols have been isolated from Ginkgo biloba
sarcotestas. Bilobal C15:1 (93) and C13:0 (94) were active against
HIV-1 PR, with IC50 values of 2.6 and 5.8 mM, respectively.
Alkylsalicylic acid C15:1 (95), C17:1 (96) and C15:0 (97) were found
to be active against HIV-1 PR, with IC50 values of 24.9, 17.8 and
10.2 mM. Compounds 95 and 96 were also found to be active
against HIV-RT, with IC50 values of 33.7 and 58.5 mM, respec-
tively. Bilobals possessing two hydroxy groups at the meta
position were found to be more active against HIV-1 PR than
alkylsalisylic acids (one hydroxyl and one carboxylic acid) or
carbanols (only one hydroxyl group). Alkylsalicylic acids with
one carboxylic acid group exhibited moderate activity against
RNase from HIV-1 RT.65
2.3 Terpenes
Lee et al. have recently reviewed plant-derived anti-HIV tri-
terpenes.66 We have included here those terpenes that were not
reported in this earlier review. The structures of anti-HIV
terpenes are shown in Figs. 7 and 8.
Fig. 8 Known terpenes discovered to be anti-HIV agents.
Nat. Prod. Rep., 2010, 27, 1781–1800 | 1789

2.3.1 Newly discovered anti-HIV terpenes. Three new tri-
terpenoids, (20R)-20,25-epoxydammaran-2-en-6a,12b-diol (98),
(20R)-20,25-epoxy-3-methyl-28-nordammaran-2-en-6a,12b-diol
(99) and isodehydroprotopanaxatriol (100) were isolated from an
acidic hydrolysate of Panax ginseng. These compounds showed
inhibitory activity against HIV-1 PR, with IC50 values of 10.5,
10.3 and 12.3 mM, respectively.67 Two new lupanes, 2a-acetoxy-
3b-hydroxy-19b-H-lup-20(29)-en-28-oic acid (2-acetoxy-
alphitolic acid) (101) and 2a-hydroxy-3b-acetoxy-19b-H-lup-
20(29)-en-28-oic acid (3-acetoxyalphitolic acid) (102) were iso-
lated from the EtOAc extract of resins of Garcinia hanburyi
collected from Thailand. Lupanes 101 and 102 showed anti-HIV-
1 activity in the syncytium assay with EC50 values of 5.6 and 6.0
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mg/mL and SI values of 2.8 and 3.3, respectively. They also dis-
played anti-HIV-1 RT inhibitory activity, with IC50 values of
116.9 and 16.3 mg/mL, respectively.68
Schilancidilactone A (103), a novel tetranortriterpenoid
possessing an unprecedented C26 skeleton (and thus representing
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a new class of Schisandra nortriterpenoids), was isolated from the
70% aqueous acetone extract of stems of Schisandra lancifolia.
Compound 103 showed anti-HIV-1 activity, with an EC50 of 8.51
mg/mL and an SI >23.5.69
Highly functionalized ingol diterpene 8-methoxyingol-7,12-
diacetate-3-phenylacetate (104) was isolated from Euphorbia
officinarum latex collected from plants on the North Atlantic
coast of Morocco at Agadir. Compound 104 caused cell-cycle
arrest and HIV-1-LTR promoter activation through a common
signalling pathway in a concentration-dependent manner, and
was proposed as a new lead for the development of therapies
against HIV-1 latency.70
The sesquiterpene isolitseane B (105) was isolated from the
chloroform extract of the leaves and twigs of Litesea verticillata
collected from Vietnam. Compound 105 was found to inhibit
HIV-1 replication, with an IC50 of 38.1 mM and an SI of 3; the 6a
isomer was found to be inactive.71
2.3.2 Known terpenes discovered to be anti-HIV agents. Two
cucurbitacins, 25-O-acetyl-23, 24-dihydrocucurbitacin F and
dihydrocucurbitacin F, named hemslecins A (106) and B (107)
respectively, were isolated from tubers of a Chinese medicinal
plant, Hemsleya jinfushanensis. These compounds were earlier
isolated from many plants of this genus, which are used to cure
inflammatory diseases such as enteritis, diarrhoea and bronchitis.
Both 106 and 107 inhibited syncytia formation with EC50 values
of 3.09 and 2.53 mg/mL, and p24 antigen production with EC50
values of 3.97 and 18.90 mg/mL, respectively. They also inhibited
fusion between HIV-1-infected H9 cells and C8166 cells with
EC50 values of 1.8 and 11.9 mg/mL. Although these compounds
showed slight interaction with HIV-1 IN, they did not inhibit
HIV-1 RT, PR, or NCp7 zinc ejection. It was concluded that 106
and 107 block or compete for the binding site of HIV-1, or
modulate the molecular expression which is necessary for HIV-1
binding to reduce entry of the virus. The acetyl group was found
to enhance the anti-HIV-1 activities of this type of compounds.72
3,4-Seco-(24Z)-cycloart-4(28),24-diene-3,26-dioic acid 26-methyl
ester (108) was isolated from Illicium verum, and inhibited HIV-
1IIIB with an EC50 of 5.1 mM.73
Labdane diterpenes forskolin (109) and 1-deoxyforskolin (110)
were isolated from the chloroform extract of Coleus forskohlii
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and showed activity against HIVNL4-3 at 5 mg/mL. Forskolin, the
major constituent of this plant, is reported to show various
biological activities, including cardiotonic, antihypertensive, and
adenylcyclase activator.74 Sesquiterpenes oxyphyllendiol B (111)
and 1,2,3,4-tetrahydro-2,5-dimethyl-8-(1-methylethyl)-1,2-
naphthalenediol (112) were isolated from Litesea verticillata.
Compunds 111 and 112 were found to inhibit HIV-1 replication
in the GFP-based cell line HOG.R5, with IC50 values of 54.6 and
91.0 mM, respectively.71
2.4 Proteins and polysaccharides
Aqueous and 80% ethanolic extracts of 20 Thai medicinal plants
used to treat AIDS were tested for their HIV-1 RT inhibitory
activity. The aqueous extracts of Ipomoea carnea subsp. fistulosa
(aerial parts), Vitex glabrata (branches), Vitex trifolia (aerial
parts), Vitex negundo (aerial parts), Canna indica (rhizomes), and
Justicia gendarussa (aerial parts) showed HIV-1 RT inhibition in
a radiometric assay at 200 mg/mL. Bioactivity-guided fraction-
ation of an aqueous extract of Canna indica rhizomes (IC50
22.6 mg/mL) yielded two proteins, Cip31 (31 kDa) and Cip14
(14 kDa). Both the proteins were found to be active against HIV-
1 RT, with IC50 values of 17.4 and 19.2 mg/mL, respectively.75
Juncin, an 18.9 kDa protein, was isolated from seeds of Brassica
juncea var. integrifolia, and showed activity against HIV-1 RT
with an IC50 of 4.5 mM.76 A 38 kDa protein was isolated from
fresh Capparis spinosa melon seeds. It exhibited partial similarity
in an N-terminal amino acid sequence with imidazole glycerol
phosphate synthase, and inhibited HIV-1 RT with an IC50 of
0.23 mM. The mechanism of inhibition may be protein–protein
interaction; HIV-1 PR is reported to inhibit the homologous
retroviral RT in a similar fashion.77 Griffithsin, a novel anti-HIV
protein with 121 amino acids and a molecular weight of
13 kDa, was isolated from an aqueous extract of the red alga
Griffithsia sp. collected from the waters of New Zealand. Grif-
fithsin displayed antiviral activity against laboratory strains and
primary isolates of T- and M-tropic HIV-1, with EC50 values
ranging from 0.043 to 0.63 nM. Griffithsin also stopped cell
fusion and transmission of HIV-1 infection at similar concen-
trations.78
Several cyclotides, macrocyclic plant peptides containing three
disulfide bonds, have been reported as anti-HIV agents. Various
cyclotides have been isolated from Viola yedoensis, a Chinese
herb. The isolated cyclotides kalata B1, varv E, and cycloviolacin
Y1, Y4 and Y5 were active in an XTT-based anti-HIV assay in
the range 0.04–1.2 mM. The most active, cycloviolacin Y5, was
also the most hydrophobic peptide of all the isolated compounds.
Therefore, a positive correlation between the hydrophobicity of
cyclotides and anti-HIV activity was inferred.79
Polysaccharide fractions TK-V2, 3 and 4, containing spirulan-
like substances, were isolated from Arthrospira platensis. TK-V2
contained a high quantity of carbohydrates while TK-V3 was
composed of 41% carbohydrates and 57% proteins. TK-V4 was
the soluble part of precipitated TK-V3, containing 45% carbo-
hydrates. All fractions showed an inhibitory effect against HIV-
1NL4-3 at 90 mg/mL. TK-V3 and 4 showed IC50 values of 3.2 and
2.1 mg/mL, respectively.80
A dimeric 64 kDa meliobiose-binding lectin was isolated from
the seeds of Bauhinia variegata. This lectin inhibited HIV-1
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reverse transcriptase with an IC50 of 1.02 mM.81 The mannose-

specific lectins from Galanthus nivalis (GNA) and a Hippeastrum
hybrid (HHA) occur as tetramers of four 12.5 kDa peptides.
Further, they were not glycosylated, and were present as
a mixture of isomers. Both lectins exhibited antiviral activity
against HIV-1 and HIV-2 in CEM cell cultures, with EC50 values
between 0.12 and 1.2 mg/mL for GNA and between 0.18 and
0.70 mg/mL for HHA, depending on the nature of the virus
strain.82 A 60 kDa lectin from seeds of the autumn purple bean
(Phaseolus vulgaris) was isolated and the sequence of its first 20
N-terminal amino acids was identified as ANEIYFSFQRF-
NETNLILQR. The lectin inhibited HIV-1 RT with an IC50 of
1.8 mM.83 BanLec, a jacalin-related lectin isolated from the fruit
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of the banana Musa acuminata, exists as a dimer with a molecular

mass of 30 kDa. BanLec showed anti-HIV activity, with IC50
values in the low nanomolar to picomolar range in different
assays with various isolates. BanLec inhibited HIV-1 infection by
binding to the glycosylated viral envelope and blocking cellular
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entry.84

2.5 Miscellaneous compounds

A new benzofuranone lactone, 5-hydroxy-3-(propan-2-ylidene)-
7-(3,7,11,15-tetramethylhexa-deca-2,6,10,11-tetraenyl)-2(3H)-
benzofuran, named rhuscholide A (113), was isolated from the
ethanolic extract of stems of Rhus chinensis. It demonstrated
anti-HIV-1 activity, with an EC50 of 1.62 mM and a TI of 42.4. A
similar compound lacking the 3-(propan-2-ylidene) group was
found to be less active and more toxic (EC50 3.7 and TI 3.2) in
a cell-based assay.85 These compounds did not inhibit HIV-RT
and PR in an enzyme-based assay. Furthermore, in a co-culti-
vation assay, these compounds did not inhibit cell–cell fusion.
Thus, it was proposed that this type of compound might target
the late steps of the HIV life-cycle.86 Butanolides 3-epi-litsenolide
D2 (114) and litseabutenolide (115) were isolated from the leaves
and twigs of Litesea verticillata, and were found to inhibit HIV-1
replication in the GFP-based cell line HOG.R5 with IC50 values
of 9.9 and 40.3 mM.71 ()-Illicinone A (116) was isolated from
Illicium verum, and inhibited HIV-1IIIB with an EC50 of
16.0 mM.73
5-Hydroxymethyl-(2,20 :50 ,200 )terthienyl tiglate (117),
5-hydroxymethyl-(2,20 :50 ,200 )terthienyl agelate (118), 5-hydroxy-
methyl-(2,20 :50 ,200 )terthienyl acetate (119) and ecliptal (120) were
isolated from the whole plant extract of Eclipta prostrata,
a perennial herb which grows widely throughout tropical areas of
Asia. Compounds 117, 119 and 120 were active against HIV-1
PR with IC50 values of 58.3, 93.7 and 83.3 mM. Compound 117
which is in the cis form, exhibited activity, whereas 118, which is
in the trans form, was inactive (IC50 >100 mM) against HIV-1
PR. This suggests that the geometry of double bond plays an
important role in activity.50 The structures are shown in Fig. 9.
3 Natural products of microbial origin
Singh et al. have reviewed HIV integrase inhibitors of mircobial
origin.7 The structures of microorganism-derived anti-HIV
natural products are shown in Figs. 10 and 11.
Fig. 9 Miscellaneous anti-HIV compounds.
3.1 Newly discovered anti-HIV agents of microbial origin
Cytochalasins are a group of fungal secondary metabolites with
various biological activities. Alachalasin A (121) was isolated
from the cultures of an isolate of Stachybotrys charatum collected
from a Tibetan glacier. Compound 121 showed inhibitory effect
on HIV-1LAI replication in C8166 cells, with an EC50 of
8.01 mM.87 Cytochalasins show antimicrobial activity, inhibition
of glucose transport, regulate plant growth and are known to
inhibit angiogenesis.88 Pestalofones A (122), B (123) and E (124)
are new cyclohexanone derivatives isolated from cultures of the
plant endophytic fungus Pestalotiopsis fici and displayed inhib-
itory effects on HIV-1 replication in C8166 cells, with EC50
values of 90.4, 64.0 and 93.7 mM, respectively. All three
compounds showed CC50 values of >200 mM.89
Three compounds, 10-methoxydihydrofuscin (125), fuscinarin
(126) and fuscin (127), were isolated from the soil fungus
Oidiodendron griseum. Compound 127 was isolated previously
from various species of genus Oidiodendron. Compounds 125–
127 showed competitive inhibition of the binding of macrophage
inflammatory protein (MIP)-1a to human CCR5 with IC50
values of 154, 80 and 21 mM, respectively, in the scintillation
proximity binding assay. The higher activity of 127 compared to
125 and 126 was ascribed to the presence of multiple reactive sites
in the isochromene-5,9-dione unit.90
Azaphilones phomoeuphorbin A (128) and phomoeuphorbin
C (129), isolated from the cultures of Phomopsis euphorbiae, an
endophytic fungus isolated from Trewia nudiflora, were tested
for in vitro inhibitory effects against HIV replication in C8166
cells. Compounds 128 and 129 showed anti-HIV activities with

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Fig. 10 Newly discovered anti-HIV agents of microbial origin.
EC50 values of 79 mg/mL and 71 mg/mL and cytotoxicity
against C8166 cells at CC50 >200 mg/mL. However, the ther-
apeutic index of >2.5 was not favourable for their future
development.91 Helotialins A–C (130–132) were isolated as new
azaphilones from the cultures of an ascomycete of the order
Helotiales. Compounds 130 and 131 displayed inhibitory
effects on HIV-1 replication in C8166 cells with EC50 values of
8.01 and 27.9 mM, respectively. Compound 132 was inactive. It
was observed that the C-13–C-14 olefin might be important
for anti-HIV activity.92
A culture of the fungus Mollisia sp., which was isolated from
the dead bark of Tsuga canadensis collected on Hamburg
Mountain, NJ, USA, by Merck Research Laboratories yielded
ophiobolin C (133), while another fungal culture of Xylaria sp.
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isolated from a fruiting body collected in Puerto Rico yielded
19,20-epoxycytochalasin Q (134). Both compounds exhibited
binding of 125I-gp120 to human CCR5 with an IC50 of 40 and
60 mM, respectively.12 Two CCR-5 inhibitors, Sch 210971 (135)
and Sch 210972 (136) were isolated from the fungus Chaetomium
globosum. Compounds 135 and 136 had IC50 values of 1.2 mM
and 79 nM respectively in the CCR-5 receptor binding assay.93
Variecolin (137) and variecolol (138), isolated from a 1 : 1
CH2Cl2–CH3OH extract of the fungus Emericella aurantio-
brunneai, had IC50 values of 9 and 32 mM respectively for binding
to human CCR5 in a scintillation proximity assay. Differences in
IC50 values suggested that the C-4 ketone and the C-20 aldehyde
might be important for activity against the human CCR5
receptor.94
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Fig. 11 Known microorganism-derived natural products discovered to be anti-HIV agents.

3.2 Known microbial metabolites discovered to be anti-HIV fuscoatra. SCH 642305 (142) was produced from fungus Septo-
agents fusidium sp. collected from Puerto Rico, having earlier been
reported from Penicillium verrucosum. Compounds 140–142
Hinnuliquinone (139) a C2-symmetric bis-indolyl quinone, was
inhibited HIV-1 Tat transactivation with IC50 values of 0.027, 5.0
isolated from the fungus on the leaves of Quercus coccifera
and 1.0 mM. Compound 142 was more toxic to the Jurkat cell line
collected in Spain. Hinnuliquinone, originally isolated from
than 140 and 141 (IC50 4.10 mM vs. $100 mM). Similar molecules
Nodulisporium hinnuleum, inhibited both the wild-type and
lacking a keto group in the macrocyclic ring were found to be
a clinically resistant (A44) strain of HIV-1 PR, with IC50 values
inactive. The sesquiterpenoids (+)-sporogen AO1 (143) and
of 2.5 and 1.8 mM, respectively. Simultaneously, some related
(+)-petasol (144) were isolated from EtCOMe extract of Pen-
compounds having an isomeric quinone moiety or benzoquinone
icillum sp. from a soil sample. Compound 143 was first isolated
were screened for anti-HIV PR activity, and were found to be
from Hansfordia pulvinata and 144 was isolated from a whole-
inactive. It was therefore assumed that the activity was not
plant extract of Petasites fragrans. These compounds inhibited
simply because of the quinone moiety, but some other structural
HIV-1 Tat transactivation, with IC50 values of 15.8 and 46.2 mM,
features of the molecule might be responsible for the activity.95
respectively.
Merck laboratories have identified several natural products
Echinomycin (145; quinomycin A) is a quinoxiline bicyclic
active against HIV-1 by using a HIV-1 Tat protein dependent in
octadepsipeptide produced by a Streptomyces sp. collected from
vitro cell-based assay. The identified compounds include the
Japan. It was reported to be a HIV-1 RT inhibitor with an IC50
macrocyclic lactones monorden (140), monocillin IV (141) and
of 700 mM.96 Sulfonium salts of echinomycin showed potent
SCH 642305 (142), the sesquiterpenoids sporogen AO1 (143) and
antitumour activity, while similar octadepsipeptides show anti-
petasol (144), the bicyclic octadepsipeptides echinomycin (145)
bacterial activity.97 UK-63598 (146) is a quinoline bicyclic octa-
and UK-63598 (146), and a cyclic heptapeptide, ternatin (147).96
depsipeptide isolated from the EtCOMe extract of
Monorden (140), also known as radicicol, and monocillin IV
Dactylosporangium sp., earlier reported from Streptomyces
(141) were produced by Penicillium sp. Compound 140 was
braegenesis. Compound 146 inhibited Tat transactivation with
originally isolated from Monosporium bonorden and subse-
an IC50 of 0.2 nM and was non-cytotoxic to Jurkat cells up to
quently it was reported from several fungal species. Monocillin
0.05 mM, whereas 145 was slightly less active, with an IC50 of
IV (141) was reported from Monocillum nordinii and Humicola
1.5 nM and a CC50 of 0.025 mM. Ternatin (147) was isolated

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from EtCOMe extract of the fermented broth of a fungus
belonging to the Xylariaceae family found on the leaves of
Quercus ilex, and inhibited Tat transactivation with an IC50 of
0.3 mM.96
4 Natural products of marine origin
A new water-soluble flavonoid glycoside, luteolin 7-b-D-gluco-
pyranosyl-200 -sulfate, named thalassiolin A (148), was isolated
from the Caribbean sea grass Thalassia testudinum. It inhibited
HIV-1 integrase terminal cleavage and strand transfer activities
with IC50 values of 2.1 and 0.4 mM, respectively, and also showed
antiviral activity in an MT-2 cell based assay, with an IC50 of
Published on 25 October 2010 on http://pubs.rsc.org | doi:10.1039/C0NP00025F
27 mM. Molecular docking studies in HIV-1 IN suggested that
148 can be accommodated near the catalytic site of the enzyme,
the binding mode overlapping with the location of the ligand
(5CITEP). The tetrazole ring of 5CITEP occupies the same
binding position as the sulfated glucose of 148, and the orien-
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tations of the keto–enol and chloroindole moieties of 5CITEP
show similarities with the benzopyranone binding position.98
Diphlorethohydroxycarmalol (149), a carmalol derivative
isolated from the marine brown alga Ishige okamurae collected
from the coasts of Korea, exhibited inhibitory effects on HIV-1
reverse transcriptase and integrase, with IC50 values of 9.1 mM
and 25.2 mM, respectively. However, it did not show any inhib-
itory activity against HIV-1 PR.99 Phloroglucinol derivative 6,60 -
bieckol (150) was isolated from Korean brown alga Ecklonia
cava, and is naturally abundant in the genus. Kim et al. showed
that 150 had inhibitory activity against HIV-1-induced syncytia
formation (EC50 1.72 mM), lytic effects (EC50 1.23 mM), and viral
p24 antigen production (EC50 1.26 mM). It also selectively
inhibited the activity of HIV-1 RT, with an EC50 of 1.07 mM.100
Two diterpenes, (6R)-6-hydroxydichotoma-3,14-diene-1,17-
dial (151) and (6R)-6-acetoxydichotoma-3,14-diene-1,17-dial
(152) were isolated from the Brazilian brown alga Dictyota
menstrualis. Compounds 151 and 152 inhibited virus replication
with EC50 of 40 and 70 mM, respectively. Moreover, 151 and 152
reduced HIV-1 RT activity, with IC50 values of 10 and 35 mM,101
and specifically inhibited RNA-dependent DNA polymerase,
with Ki values of 10 and 35 mM. Neither compound inhibited
DNA-dependent DNA-polymerase102 or affected cell viability or
proliferation, and therefore the synthesis of proviral DNA was
considered as a specific target for the anti-HIV-1 activity of these
diterpenes.101
Dolabellane diterpene 8,10,18-trihydroxy-2,6-dolabelladiene
(153) isolated from the alga Dictyota pfaffii, inhibited HIV-1
infection in PBMCs, with an EC50 of 8.4 mM, and in macro-
phages, with an EC50 of 1.7 mM. It also showed inhibition of
HIV-1 RT, with an IC50 of 16.5 mM.103,104
A new furanoterpene, designated dehydrofurodendin, was
isolated from Madagascan sponges of the genus Lendenfeldia.
Dehydrofurodendin (154) inhibited HIV-1 RT-associated DNA
polymerase activities, with IC50 values of 3.2 and 5.6 mM for
RNA- and DNA-directed DNA polymerase functions, respec-
tively. It also inhibited the RNase H activity of HIV-1 RT, with
an IC50 value of 29.5 mM.105
Neamphamide A (155), a cyclic depsipeptide containing 11
amino acid residues, was isolated from a Papua New Guinea
collection of the marine sponge Neamphius huxleyi, and was
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found to inhibit the cytopathic effect of HIV-1, with an EC50 of
28 nM in a cell-based XTT assay. Direct cytotoxicity of 155
against the host cells was observed, with a TC50 of 260 nM.106
Some other cyclic depsipeptides, mirabamides A–D, were iso-
lated from marine sponge material collected from Nama Island,
Micronesia. Mirabamides A, C and D (156–158) inhibited HIV-1
in a neutralization assay in two different viral strains (HXB2 and
SF162) with IC50 values of 140 and 40 nM, 144 nM and 1 mM,
and 189 nM and 1.3 mM, respectively. They also inhibited HIV-1-
mediated cell fusion against LAV, with IC50 values of 41 nM, 1.3
and 3.9 mM, respectively. These results suggested that these
compounds might act at the early stages of HIV-1 entry. Mir-
abamide A (156), which contains the rhamnosylated b-OMe Tyr
residue, was observed to be the most potent of all the isolated
compounds. Earlier, it had been suggested that the b-OMe Tyr
residue is critical for the anti-HIV activity of a structurally
similar compound, theopapuamide.107 However, it was found
that in the case of mirabamides, the b-OMe Tyr was not the only
essential residue for activity, because b-OMe Tyr with substitu-
tion at the 40 -position can be tolerated with no adverse effects on
antiviral activity, and the presence of a free 40 -hydroxyl on the
Tyr unit was not essential. Yet another important finding was
that compound 156 was equipotent to papuamide A (which
contains a b-OMe Tyr), while theopapauamide (which lacks
a b-OMe Tyr residue) was inactive. The b-OMe Tyr residue
provides a specific conformation required for binding to the
target protein involved in HIV-1 entry.108 Homophymine A
(159), a cyclodepsipeptide isolated from the marine sponge
Homophymia sp. collected in New Caledonia, effectively inhibi-
ted HIV-1IIIb infection with an IC50 of 75 nM. Direct cytotoxicity
of 159 against the host cells was observed, with a TC50 of
1.19 mM.109 The structures are shown in Fig. 12.
5 Advanced-stage molecules – where are they now?
In the past, several anti-HIV natural products have proceeded to
various stages of drug development. The structures of these
molecules is given in Fig. 13, and their present status is provided
in Table 1.
Calanolide was the most active anti-HIV-1 compound isolated
in the NCI-led discovery program from the rainforest tree
Calophyllum lanigerum.110 (+)-Calanolide A (160) inhibited
various laboratory strains of HIV-1 with EC50 0.10–0.17 mM. It
selectively inhibited recombinant HIV-1 RT, but not cellular
DNA polymerases or HIV-2 RT.111 Mechanism-of-action studies
revealed that it bound near the active site of the enzyme HIV-1
RT and interfered with dNTP binding. It inhibited HIV-1 RT in
a synergistic manner with nevirapine, suggesting that it was
different than the general class of nucleoside RT inhibitors.112 A
single amino acid change in HIV-1 RT (V106A or Y181C) lead to
significant resistance to nevirapine, but sensitivity to calanolide
A was unaffected. In contrast, the Y188H mutant had 30-fold
increased resistance to calanolide A, while nevirapine did not
show any resistance.113
The efficacy of 160 was demonstrated in vivo in a hollow-fiber
mouse model following oral or parenteral administration.114 The
pharmacokinetic data after i.v. administration of calanolide A
and dihydrocalanolide A were almost the same (AUC (area
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Fig. 12 Anti-HIV compounds of marine origin.
under the curve), t1/2, clearance). Dihydrocalanolide A showed
better oral bioavailability.115
Phase I studies in 47 healthy volunteers using 160 encapsulated
in a sesame oil-based soft gelatin capsule demonstrated its safety
and favorable pharmacokinetic profile. No serious adverse side-
effects were reported; the majority of all adverse events were
ranked as grade 1 (mild), and included symptoms of dizziness,
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taste perversion, headache, eructation, and nausea.116,117 The
latest information available states that (+)-calanolide A is in
Phase IB trials for evaluation of the safety, pharmacokinetics,
and effects in HIV-positive patients.118
Bevirimat, 3-O-(30 ,30 -dimethylsuccinyl)betulinic acid (161),
showed a potent activity in acutely infected H9 lymphocytes with
an EC50 value of <0.35 nM.119 The design of 161 was based on
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Fig. 13 Advanced-stage molecules.
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Table 1 Advanced-stage anti-HIV natural products
Molecule Stage
Calanolide A Phase I
Bevirimat Phase IIb
PA-1050040 Phase I
DCK Preclinical
DCQA Preclinical
Dextran sulfate Phase I
Curcumin Preclinical
Cyanovirin A Preclinical
HIV-1 entry inhibitors with the same skeleton (betulinic acid).
Bevirimat did not affect viral attachment or fusion, RT, PR or
IN function, but instead acted at a late stage in HIV replica-
tion.120 Studies to understand its mechanism of action revealed
that 161 dose-dependently inhibited the processing of the viral
Gag polyprotein at a specific step, namely the conversion of the
capsid precursor p25 to mature capsid protein p24.121 This
inhibition led to the production of non-infectious and immature
viral particles, resulting in a new class of anti-HIV agents known
as maturation inhibitors.66 Based on the potent in vitro and in
vivo antiviral activity along with supportive preclinical data, 161
was advanced into clinical testing and was granted ‘Fast-Track’
New Drug Development status by the US FDA in December
2004.120 As of 2008, it was in Phase IIb clinical trials.66 Bevirimat
offers a favorable pharmacokinetic profile in healthy humans
and HIV-infected patients after a single oral administration.122,123
Bevirimat was well absorbed after oral administration, and peak
plasma concentrations were achieved in 1–3 h. A long half-life of
60–80 h may facilitate once-daily dosing. It is not oxidatively
metabolized, does not inhibit cytochrome P450,122 and is elimi-
nated by hepatic glucuronidation.124
PA1050040, an analog of bevirimat, acts as a maturation
inhibitor and is a potent inhibitor of virus replication in primary
and prototypic HIV-1 isolates, with an EC50 of 15 nM. It was
also active against the bevirimat-resistant wild-type isolate
L363M, and isolates resistant to approved ARVs. It is orally
bioavailable in rats. The latest information available indicates
that it has entered Phase I clinical trials for single-dose testing.125
1796 | Nat. Prod. Rep., 2010, 27, 1781–1800
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DCK, 30 R,40 R-di-O-()-camphanoyl-(+)-cis-khellactone
(162), has been synthesized from suksdorfin by replacing the 30 -
acetyl and 40 -isovaleryl groups with camphanoyl esters. It
showed potent activity against HIV-1IIIB compared to suksdor-
fin, with an EC50 of 0.049 mM.126 SAR studies suggested that
a methyl group at C-3, C-4, or C-5 increased anti-HIV activity.
4-Methyl-DCK was 10 times more active than DCK (EC50
0.0059 mM).127 Preliminary studies on the mechanism of action
showed that it selectively inhibited virus replication in HIV-1-
infected cells, but did not inhibit the activity of three major HIV
enzymes, HIV-RT, PR, and IN. The studies indicated that 162
inhibited HIV after viral entry but prior to viral DNA integra-
tion.4 DCK inhibits HIV-RT at a later step than that affected by
AZT and other clinically approved RT inhibitors. Studies on the
analog hydroxymethyl DCK suggested that it inhibited the
production of double-stranded viral DNA from the single-
stranded DNA intermediate.120 The novel mechanism of action
of DCK is known as strand-transfer inhibition.128 Several new
analogs of DCK have been found to be more active.4,120
Several dicaffeoylquinic acids (DCQAs) were found to inhibit
HIV-1 replication at 1–6 mM in T-cell lines, whereas they were
non-cytotoxic up to 120 mM. A mechanism-based study sug-
gested that the compounds inhibited HIV-1 IN. Molecular
modelling of DCQAs with the core catalytic domain of HIV-1
IN indicated that the inhibitors occupy a groove within the
catalytic site.129 1,5-DCQA (163), another member of the
DCQA group, was found to have low bioavailability and
remained intact in rat plasma following oral administration.
Twenty-two metabolites were found in the matrices of rats,
suggesting that the extensive metabolism might be one factor
leading to the low bioavailability of 163. Three major metabo-
lites of 163 also inhibited HIV replication in MT-4 cells, with
IC50 values of 25, 25 and 46 mM respectively, making an
important contribution to the anti-HIV activity.130 In another
study, following a single intravenous administration of 163, it
disappeared rapidly from plasma with a mean terminal half-life
(t1/2) of 1.40 h. A total of 15 metabolites along with the parent
molecule were observed in rat urine. The quick urinary excre-
tion and extensive metabolism of 163 result in its rapid elimi-
nation from the system.131
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The sulfated polysaccharide dextran sulfate inhibited the repli-
cation of HIV-1 with an IC50 of 9.1 mg/mL in MT-4 cells. Dextran
sulfates with molecular weights of 5000, 8000 and 500 000 showed
a similar inhibitory effect on HIV-1 replication. They also showed
an HIV-1-RT inhibitory effect, with IC50 values of 41.9, 66.2 and
17.9 mg/mL, respectively, and interfere with virus adsorption.132
These sulfated polysaccharides are also potent inhibitors of other
enveloped viruses such as HSV, cytomegalovirus and vesicular
stomatitis virus,133 and also show strong anti-coagulant proper-
ties.132 Unfortunately, clinical trials of oral dextran sulfate in HIV-
infected patients were terminated because of poor absorption of the
drug, poor central nervous system penetration and short half-life.134
Furthermore, parenteral dextran sulfate did not lower circulating
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HIV antigen levels, but instead increased the circulating antigen.
The dextran sulfate infusion was also associated with significant
toxicity, especially thrombocytopenia. As a result, further clinical
development of dextran sulfate as a therapy for symptomatic HIV
infection was not warranted, and the authors advised caution in the
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clinical evaluation of other polysulfated polyanions.135
Curcumin (164) is the yellow pigment of turmeric (Curcuma
longa), a widely used spice and food coloring agent. Curcumin is
reported to show antitumor, anti-inflammatory and many other
biological activities. It also plays a role in the prevention of
neurological, cardiovascular and metabolic disorders.136 Curcu-
min showed HIV-1 IN inhibition, with an IC50 of 40 mM for strand
transfer reaction. HIV-1 IN inhibition by curcumin may result
from coordination of its hydroxyl groups to the acidic residues in
the active site, but it was suggested that the inhibition may involve
multiple mechanisms.137 It was also reported that it inhibits p24
antigen production and Tat-mediated transcription at higher
concentration.138 There have been almost 2500 in vitro and in vivo
studies on curcumin for diverse activities, and this has generated
enough interest for clinical trials in cancer and inflammation. One
of the most important limitations of curcumin is its low bioavail-
ability, which is due to its hydrophobic nature, and numerous
approaches have been undertaken to enhance its bioavailability.136
The safety profile and pharmacokinetic data are well established,
and it is currently in clinical trials for AIDS patients.137
Cyanovirin-N, an 11 kDa protein containing 101 amino acids,
was isolated from extracts of the cultured cyanobacterium
Nostoc ellipsosporum. It irreversibly inactivates laboratory
strains and primary isolates of HIV-1 and HIV-2 in low nano-
molar concentrations. It also inhibits cell-to-cell fusion and
transmission of HIV-1 infection.139 Cyanovirin-N binds to viral
envelope glycoproteins, and interferes with viral interactions
with target cell receptors that are essential for viral entry and cell-
to-cell fusion, targeting N-linked high-mannose oligosaccharides
on the viral envelope.140 Intravenous administration of cyano-
virin-N resulted in rapid removal by renal filtration. It has a high
potential for use as a systemic compound to control viral load or
in the development of microbiocides to prevent primary viral
infection. It was suggested that PEGylated cyanovirin-N would
be helpful to overcome problems associated with the use of
proteins. These formulations are presently in preclinical phase.141
6 Conclusions and future perspectives
The basic biological processes in the HIV life-cycle are now well-
established, and it is possible to find natural compounds
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targeting specific steps in the viral life-cycle. In this review, we
have provided an overview of various natural products isolated
from diverse sources and acting at different targets/steps of the
life-cycle, dissecting the work done over the last seven years into
two broad categories, i.e. new natural products showing anti-
HIV activity and previously known natural products that were
evaluated for this activity. As can be observed, during the period
covered in this review, fewer new natural products were discov-
ered as having anti-HIV potential compared to the much larger
number of previously known natural products found to be active
in various assays to assess anti-HIV potential. Therefore,
a multidirectional approach needs to be continued for discovery
of anti-HIV drugs from natural sources. This approach should
include finding new natural products, evaluating known natural
products having structural similarity with known anti-HIV
compounds, semi-synthetic modifications of anti-HIV natural
products in order to enhance their activity, and preparation of
composite extracts active in anti-HIV assays. Here, it becomes
important to emphasize the development of alternative therapies
using medicinal plants showing anti-HIV activities. Extracts
from plants showing anti-HIV activity should be profiled using
techniques such as HPLC–PDA and LC–MS for complete che-
moprofiling to give standardized extracts that can be used as
herbal products. Natural products have served as important
leads for the discovery of more potent molecules – for example,
bevirimat and DCK were designed on the basis of anti-HIV
activity of naturally occurring betulinic acid and suksdorfin,
respectively. The possibility of finding new molecules based on
natural products acting in the nanomolar/picomolar range need
to be explored to find effective drugs against HIV. Although very
few natural products (such as clusianone, propolone A, the lectin
BanLec, and mirabamide A) have shown anti-HIV activity in the
nanomolar/picomolar range, further efforts are warranted to find
new lead molecules for HIV/AIDS from natural sources.
7 Acknowledgement
We thank the Director of NIPER for support.
8 References
1 AIDS Epidemic Update 2009, UNAIDS, Geneva, Switzerland, 2009.
2 2008 Report on the global AIDS epidemic, UNAIDS, Geneva,
Switzerland, 2008.
3 W. C. Greene, Z. Debyser, Y. Ikeda, E. O. Freed, E. Stephens,
W. Yonemoto, R. W. Buckheit, J. A. Esteg and T. Cihlar,
Antiviral Res., 2008, 80, 251–265.
4 D. Yu, M. Suzuki, L. Xie, S. L. Morris-Natschke and K. H. Lee,
Med. Res. Rev., 2003, 23, 322–345.
5 R. S. B. Goncalves, M. L. Ferreira, M. T. Ventura, M. A. Peralta
and S. M. V. N. De, Rev. Fitos, 2006, 2, 74–81.
6 J. Nian and J. Dong, Zhongguo Yaoxue Zazhi, 2005, 40, 571–573.
7 S. B. Singh, F. Pelaez, D. J. Hazuda and R. B. Lingham, Drugs
Future, 2005, 30, 277–299.
8 G. Dong, Y. Li, Y. Liu and G. Su, Zhongguo Haiyang Yaowu, 2009,
28, 44–53.
9 P. Cos, L. Maes, A. Vlietinck and L. Pieters, Planta Med., 2008, 74,
1323–1337.
10 J. Wang, Y. Zheng, T. Efferth, R. Wang, Y. Shen and X. Hao,
Phytochemistry, 2005, 66, 697–701.
11 M. Horiuch, C. Murakami, N. Fukamiya, D. Yu, T. H. Chen,
K. F. Bastow, D. C. Zhang, Y. Takaishi, Y. Imakura and
K. H. Lee, J. Nat. Prod., 2006, 69, 1271–1274.
Nat. Prod. Rep., 2010, 27, 1781–1800 | 1797

12 H. Jayasuriya, K. B. Herath, J. G. Ondeyka, J. D. Polishook,
G. F. Bills, A. W. Dombrowski, M. S. Springer, S. Siciliano,
L. Malkowitz, M. Sanchez, Z. Guan, S. Tiwari, D. W. Stevenson,
R. P. Borris and S. B. Singh, J. Nat. Prod., 2004, 67, 1036–1038.
13 G. Li, K. M. Haney, G. E. Kellogg and Y. Zhang, J. Chem. Inf.
Model., 2009, 49, 120–132.
14 P. W. Hsieh, F. R. Chang, K. H. Lee, T. L. Hwang, S. M. Chang and
Y. C. Wu, J. Nat. Prod., 2004, 67, 1175–1177.
15 E. M. Silva, C. C. Cirne-Santos, I. C. Frugulhetti, B. Galvao-Castro,
E. M. Saraiva, M. E. Kuehne and D. C. Bou-Habib, Planta Med.,
2004, 70, 808–812.
16 W. Zhang, Y. Ramamoorthy, R. F. Tyndale, S. D. Glick,
I. M. Maisonneuve, M. E. Kuehne and E. M. Sellers, Drug Metab.
Dispos., 2002, 30, 663–669.
17 A. K. Chakravarty, T. Sarkar, K. Masuda and K. Shiojima,
Phytochemistry, 1999, 50, 1263–1266.
Published on 25 October 2010 on http://pubs.rsc.org | doi:10.1039/C0NP00025F
18 C. Ito, M. Itoigawa, A. Sato, C. M. Hasan, M. A. Rashid,
H. Tokuda, T. Mukainaka, H. Nishino and H. Furukawa, J. Nat.
Prod., 2004, 67, 1488–1491.
19 B. Kongkathip, N. Kongkathip, A. Sunthitikawinsakul,
C. Napaswat and C. Yoosook, Phytother. Res., 2005, 19, 728–731.
20 M. J. Cheng, K. H. Lee, I. L. Tsai and I. S. Chen, Bioorg. Med.
Chem., 2005, 13, 5915–5920.
Downloaded by University of Sussex on 06 August 2012
21 R. Stadler, T. M. Kutchan and M. H. Zenk, Phytochemistry, 1989,
28, 1083–1086.
22 N. Samanani, D. K. Liscombe and P. J. Facchini, Plant J., 2004, 40,
302–313.
23 Y. Kashiwada, A. Aoshima, Y. Ikeshiro, Y. P. Chen, H. Furukawa,
M. Itoigawa, T. Fujioka, K. Mihashi, L. M. Cosentino, S. L. Morris-
Natschke and K. H. Lee, Bioorg. Med. Chem., 2005, 13, 443–448.
24 M. H. Yan, P. Cheng, Z. Y. Jiang, Y. B. Ma, X. M. Zhang,
F. X. Zhang, L. M. Yang, Y. T. Zheng and J. J. Chen, J. Nat.
Prod., 2008, 71, 760–763.
25 A. L. Otshudi, S. Apers, L. Pieters, M. Claeys, C. Pannecouque,
E. De Clercq, A. Van Zeebroeck, S. Lauwers, M. Frederich and
A. Foriers, J. Ethnopharmacol., 2005, 102, 89–94.
26 M. L. Lohombo-Ekomba, P. N. Okusa, O. Penge, C. Kabongo,
M. I. Choudhary and O. E. Kasende, J. Ethnopharmacol., 2004,
93, 331–335.
27 L. Szlavik, A. Gyuris, J. Minarovits, P. Forgo, J. Molnar and
J. Hohmann, Planta Med., 2004, 70, 871–873.
28 B. Sener, I. Orhan and J. Satayavivad, Phytother. Res., 2003, 17,
1220–1223.
29 P. L. Wu, F. W. Lin, T. S. Wu, C. S. Kuoh, K. H. Lee and S. J. Lee,
Chem. Pharm. Bull., 2004, 52, 345–349.
30 A. Saklani and S. K. Kutty, Drug Discovery Today, 2008, 13, 161–
171.
31 M. Chen, N. Kilgore, K. H. Lee and D. F. Chen, J. Nat. Prod., 2006,
69, 1697–1701.
32 Q. Z. Sun, D. F. Chen, P. L. Ding, C. M. Ma, H. Kakuda,
N. Nakamura and M. Hattori, Chem. Pharm. Bull., 2006, 54, 129–132.
33 X. N. Li, J. X. Pu, X. Du, L. M. Yang, H. M. An, C. Lei, F. He,
X. Luo, Y. T. Zheng, Y. Lu, W. L. Xiao and H. D. Sun, J. Nat.
Prod., 2009, 72, 1133–1141.
34 K. M. Meragelman, T. C. McKee and M. R. Boyd, J. Nat. Prod.,
2001, 64, 546–548.
35 T. C. McKee, H. R. Bokesch, J. L. McCormick, M. A. Rashid,
D. Spielvogel, K. R. Gustafson, M. M. Alavanja, J. H. Cardellina
and M. R. Boyd, J. Nat. Prod., 1997, 60, 431–438.
36 V. Reutrakul, N. Ningnuek, M. Pohmakotr, C. Yoosook,
C. Napaswad, J. Kasisit, T. Santisuk and P. Tuchinda, Planta
Med., 2007, 73, 683–688.
37 S. Cao, C. Rossant, S. Ng, A. D. Buss and M. S. Butler,
Phytochemistry, 2003, 64, 987–990.
38 G. Appendino, M. Ottino, N. Marquez, F. Bianchi, A. Giana,
M. Ballero, O. Sterner, B. L. Fiebich and E. Munoz, J. Nat. Prod.,
2007, 70, 608–612.
39 N. T. Dat, K. Bae, A. Wamiru, J. B. McMahon, S. F. Le Grice,
M. Bona, J. A. Beutler and Y. H. Kim, J. Nat. Prod., 2007, 70,
839–841.
40 H. J. Kim, E. J. Kim, S. H. Seo, C. G. Shin, C. Jin and Y. S. Lee, J.
Nat. Prod., 2006, 69, 600–603.
41 J. X. Pu, L. M. Yang, W. L. Xiao, R. T. Li, C. Lei, X. M. Gao,
S. X. Huang, S. H. Li, Y. T. Zheng, H. Huang and H. D. Sun,
Phytochemistry, 2008, 69, 1266–1272.
1798 | Nat. Prod. Rep., 2010, 27, 1781–1800
View Online
42 W. L. Xiao, S. X. Huang, R. R. Wang, J. L. Zhong, X. M. Gao,
F. He, J. X. Pu, Y. Lu, Y. T. Zheng, Q. T. Zheng and H. D. Sun,
Phytochemistry, 2008, 69, 2862–2866.
43 X. M. Zhang, D. F. Chen, X. J. He, S. Yang, P. Zheng and
M. H. Jiang, Acta Pharmacol. Sinica, 2000, 21, 373–376.
44 J. Zhang, L. L. Shi and Y. N. Zheng, Nat. Prod. Commun., 2010, 5,
231–234.
45 S. Y. Oh, Y. H. Kim, D. S. Bae, B. H. Um, C. H. Pan, C. Y. Kim,
H. J. Lee and J. K. Lee, Biosci., Biotechnol., Biochem., 2010, 74,
285–291.
46 X. M. Gao, J. X. Pu, S. X. Huang, L. M. Yang, H. Huang,
W. L. Xiao, Y. T. Zheng and H. D. Sun, J. Nat. Prod., 2008, 71,
558–563.
47 Y. B. Yu, H. Miyashiro, N. Nakamura, M. Hattori and J. C. Park,
Arch. Pharmacal Res., 2007, 30, 820–826.
48 J. Lee, W. K. Oh, J. S. Ahn, Y. H. Kim, J. T. Mbafor, J. Wandji and
Z. T. Fomum, Planta Med., 2009, 75, 268–270.
49 C. H. Ma, R. R. Wang, R. R. Tian, G. Ye, M. S. Fan, Y. T. Zheng
and C. G. Huang, Chem. Nat. Compd., 2009, 45, 282–285.
50 S. Tewtrakul, S. Subhadhirasakul, S. Cheenpracha and C. Karalai,
Phytother. Res., 2007, 21, 1092–1095.
51 C. C. Li, Z. X. Xie, Y. D. Zhang, J. H. Chen and Z. Yang, J. Org.
Chem., 2003, 68, 8500–8504.
52 S. Cheenpracha, C. Karalai, C. Ponglimanont, S. Subhadhirasakul
and S. Tewtrakul, Bioorg. Med. Chem., 2006, 14, 1710–1714.
53 V. Martino, J. Morales, J. J. Martinez-Irujo, M. Font, A. Monge
and J. Coussio, Phytother. Res., 2004, 18, 667–669.
54 C. C. Lin, Y. F. Hsu, T. C. Lin and H. Y. Hsu, Phytother. Res., 2001,
15, 206–212.
55 S. Jain, P. P. Yadav, V. Gill, N. Vasudeva and N. Singla, Phytochem.
Rev., 2009, 8, 491–502.
56 M. K. Reddy, S. K. Gupta, M. R. Jacob, S. I. Khan and D. Ferreira,
Planta Med., 2007, 73, 461–467.
57 J. L. Lopez-Perez, D. A. Olmedo, E. del Olmo, Y. Vasquez,
P. N. Solis, M. P. Gupta and A. San Feliciano, J. Nat. Prod.,
2005, 68, 369–373.
58 N. Marquez, R. Sancho, L. M. Bedoya, J. Alcami, J. L. Lopez-Perez,
A. S. Feliciano, B. L. Fiebich and E. Munoz, Antiviral Res., 2005, 66,
137–145.
59 L. M. Bedoya, M. Beltran, R. Sancho, D. A. Olmedo, S. Sanchez-
Palomino, E. del Olmo, J. L. L opez-Perez, E. Mu~ noz,
A. S. Feliciano and J. Alcamı, Bioorg. Med. Chem. Lett., 2005, 15,
4447–4450.
60 V. Reutrakul, N. Anantachoke, M. Pohmakotr, T. Jaipetch,
S. Sophasan, C. Yoosook, J. Kasisit, C. Napaswat, T. Santisuk
and P. Tuchinda, Planta Med., 2007, 73, 33–40.
61 V. Reutrakul, W. Chanakul, M. Pohmakotr, T. Jaipetch,
C. Yoosook, J. Kasisit, C. Napaswat, T. Santisuk, S. Prabpai,
P. Kongsaeree and P. Tuchinda, Planta Med., 2006, 72, 1433–1435.
62 A. L. Piccinelli, O. Cuesta-Rubio, M. B. Chica, N. Mahmood,
B. Pagano, M. Pavone, V. Barone and L. Rastrelli, Tetrahedron,
2005, 61, 8206–8211.
63 N. M. Ahmad, V. Rodeschini, N. S. Simpkins, S. E. Ward and
A. J. Blake, J. Org. Chem., 2007, 72, 4803–4815.
64 L. Verotta, Phytochem. Rev., 2002, 1, 389–407.
65 J. S. Lee, M. Hattori and J. Kim, Planta Med., 2008, 74, 532–534.
66 R. Y. Kuo, K. Qian, S. L. Morris-Natschke and K. H. Lee, Nat.
Prod. Rep., 2009, 26, 1321–1344.
67 Y. Wei, C.-M. Ma and M. Hattori, Phytochem. Lett., 2009, 2, 63–66.
68 V. Reutrakul, N. Anantachoke, M. Pohmakotr, T. Jaipetch,
C. Yoosook, J. Kasisit, C. Napaswa, A. Panthong, T. Santisuk,
S. Prabpai, P. Kongsaeree and P. Tuchinda, Planta Med., 2010,
76, 368–371.
69 X. Luo, Y. Chang, X. J. Zhang, J. X. Pu, X. M. Gao, Y. L. Wu,
R. R. Wang, W. L. Xiao, Y. T. Zheng and Y. Lu, Tetrahedron
Lett., 2009, 50, 5962–5964.
70 M. Daoubi, N. Marquez, N. Mazoir, A. Benharref, R. Hernandez-
Galan, E. Munoz and I. G. Collado, Bioorg. Med. Chem., 2007,
15, 4577–4584.
71 H. J. Zhang, V. H. Nguyen, M. C. Nguyen, D. D. Soejarto,
J. M. Pezzuto, H. H. Fong and G. T. Tan, Planta Med., 2005, 71,
452–457.
72 R. R. Tian, J. C. Chen, G. H. Zhang, M. H. Qiu, Y. H. Wang, L. Du,
X. Shen, N. F. Liu and Y. T. Zheng, Chin. J. Nat. Med., 2008, 6,
214–218.
This journal is ª The Royal Society of Chemistry 2010

73 W. Song, Y. Ma, X. Bai, X. Zhang, Q. Gu, Y. Zheng, J. Zhou and
J. Chen, Planta Med., 2007, 73, 372–375.
74 H. S. Bodiwala, S. Sabde, D. Mitra, K. K. Bhutani and I. P. Singh,
Nat. Prod. Commun., 2009, 4, 1173–1175.
75 W. Woradulayapinij, N. Soonthornchareonnon and C. Wiwat,
J. Ethnopharmacol., 2005, 101, 84–89.
76 X. Ye and T. B. Ng, J. Agric. Food Chem., 2009, 57, 4366–4371.
77 S. K. Lam and T. B. Ng, Phytomedicine, 2009, 16, 444–450.
78 T. Mori, B. R. O’Keefe, R. C. Sowder, 2nd, S. Bringans, R. Gardella,
S. Berg, P. Cochran, J. A. Turpin, R. W. Buckheit, Jr., J. B. McMahon
and M. R. Boyd, J. Biol. Chem., 2005, 280, 9345–9353.
79 C. K. Wang, M. L. Colgrave, K. R. Gustafson, D. C. Ireland,
U. Goransson and D. J. Craik, J. Nat. Prod., 2008, 71, 47–52.
80 S. Rechter, T. K€ onig, S. Auerochs, S. Thulke, H. Walter,
H. D€ ornenburg, C. Walter and M. Marschall, Antiviral Res., 2006,
72, 197–206.
Published on 25 October 2010 on http://pubs.rsc.org | doi:10.1039/C0NP00025F
81 P. Lin and T. B. Ng, J. Agric. Food Chem., 2008, 56, 10481–10486.
82 J. Balzarini, S. Hatse, K. Vermeire, K. Princen, S. Aquaro,
C. F. Perno, E. De Clercq, H. Egberink, G. Vanden Mooter,
W. Peumans, E. Van Damme and D. Schols, Antimicrob. Agents
Chemother., 2004, 48, 3858–3870.
83 E. F. Fang, P. Lin, J. H. Wong, S. W. Tsao and T. B. Ng, J. Agric.
Food Chem., 2010, 58, 2221–2229.
Downloaded by University of Sussex on 06 August 2012
84 M. D. Swanson, H. C. Winter, I. J. Goldstein and D. M. Markovitz,
J. Biol. Chem., 2010, 285, 8646–8655.
85 Q. Gu, R. R. Wang, X. M. Zhang, Y. H. Wang, Y. T. Zheng, J. Zhou
and J. J. Chen, Planta Med., 2007, 73, 279–282.
86 R. R. Wang, Q. Gu, Y. H. Wang, X. M. Zhang, L. M. Yang,
J. Zhou, J. J. Chen and Y. T. Zheng, J. Ethnopharmacol., 2008,
117, 249–256.
87 Y. Zhang, R. Tian, S. Liu, X. Chen, X. Liu and Y. Che, Bioorg. Med.
Chem., 2008, 16, 2627–2634.
88 A. M. Haidle and A. G. Myers, Proc. Natl. Acad. Sci. U. S. A., 2004,
101, 12048–12053.
89 L. Liu, S. Liu, X. Chen, L. Guo and Y. Che, Bioorg. Med. Chem.,
2009, 17, 606–613.
90 K. Yoganathan, C. Rossant, S. Ng, Y. Huang, M. S. Butler and
A. D. Buss, J. Nat. Prod., 2003, 66, 1116–1117.
91 B. Z. Yu, G. H. Zhang, Z. Z. Du, Y. T. Zheng, J. C. Xu and
X. D. Luo, Phytochemistry, 2008, 69, 2523–2526.
92 X.-W. Zou, B.-D. Sun, X.-L. Chen, X.-Z. Liu and Y.-S. Che, Chin. J.
Nat. Med., 2009, 7, 140–144.
93 S. W. Yang, R. Mierzwa, J. Terracciano, M. Patel, V. Gullo,
N. Wagner, B. Baroudy, M. Puar, T. M. Chan, A. T. McPhail and
M. Chu, J. Nat. Prod., 2006, 69, 1025–1028.
94 K. Yoganathan, C. Rossant, R. P. Glover, S. Cao, J. J. Vittal, S. Ng,
Y. Huang, A. D. Buss and M. S. Butler, J. Nat. Prod., 2004, 67,
1681–1684.
95 S. B. Singh, J. G. Ondeyka, N. Tsipouras, C. Ruby, V. Sardana,
M. Schulman, M. Sanchez, F. Pelaez, M. W. Stahlhut, S. Munshi,
D. B. Olsen and R. B. Lingham, Biochem. Biophys. Res. Commun.,
2004, 324, 108–113.
96 H. Jayasuriya, D. L. Zink, J. D. Polishook, G. F. Bills,
A. W. Dombrowski, O. Genilloud, F. F. Pelaez, L. Herranz,
D. Quamina, R. B. Lingham, R. Danzeizen, P. L. Graham,
J. E. Tomassini and S. B. Singh, Chem. Biodiversity, 2005, 2, 112–
122.
97 Y. B. Kim, Y. H. Kim, J. Y. Park and S. K. Kim, Bioorg. Med.
Chem. Lett., 2004, 14, 541–544.
98 D. C. Rowley, M. S. T. Hansen, D. Rhodes, C. A. Sotriffer, H. Ni,
J. A. McCammon, F. D. Bushman and W. Fenical, Bioorg. Med.
Chem., 2002, 10, 3619–3625.
99 M. J. Ahn, K. D. Yoon, C. Y. Kim, J. H. Kim, C. G. Shin and
J. Kim, Phytother. Res., 2006, 20, 711–713.
100 M. Artan, Y. Li, F. Karadeniz, S. H. Lee, M. M. Kim and S. K. Kim,
Bioorg. Med. Chem., 2008, 16, 7921–7926.
101 H. S. Pereira, L. R. Leao-Ferreira, N. Moussatche, V. L. Teixeira,
D. N. Cavalcanti, L. J. Costa, R. Diaz and I. C. Frugulhetti,
Antiviral Res., 2004, 64, 69–76.
102 H. de Souza Pereira, L. R. Leao-Ferreira, N. Moussatche,
V. L. Teixeira, D. N. Cavalcanti, L. J. da Costa, R. Diaz and
I. C. Frugulhetti, Planta Med., 2005, 71, 1019–1024.
103 J. P. Barbosa, R. C. Pereira, J. L. Abrantes, C. C. Cirne dos Santos,
M. A. Rebello, I. C. Frugulhetti and V. L. Texeira, Planta Med.,
2004, 70, 856–860.
This journal is ª The Royal Society of Chemistry 2010
View Online
104 C. C. Cirne-Santos, V. L. Teixeira, L. R. Castello-Branco,
I. C. Frugulhetti and D. C. Bou-Habib, Planta Med., 2006, 72,
295–299.
105 L. Chill, A. Rudi, M. Aknin, S. Loya, A. Hizi and Y. Kashman,
Tetrahedron, 2004, 60, 10619–10626.
106 N. Oku, K. R. Gustafson, L. K. Cartner, J. A. Wilson,
N. Shigematsu, S. Hess, L. K. Pannell, M. R. Boyd and
J. B. McMahon, J. Nat. Prod., 2004, 67, 1407–1411.
107 A. S. Ratnayake, T. S. Bugni, X. Feng, M. K. Harper, J. J. Skalicky,
K. A. Mohammed, C. D. Andjelic, L. R. Barrows and C. M. Ireland,
J. Nat. Prod., 2006, 69, 1582–1586.
108 A. Plaza, E. Gustchina, H. L. Baker, M. Kelly and C. A. Bewley,
J. Nat. Prod., 2007, 70, 1753–1760.
109 A. Zampella, V. Sepe, P. Luciano, F. Bellotta, M. C. Monti,
M. V. D’Auria, T. Jepsen, S. Petek, M. T. Adeline, O. Laprevote,
A. M. Aubertin, C. Debitus, C. Poupat and A. Ahond, J. Org.
Chem., 2008, 73, 5319–5327.
110 Y. Kashman, K. R. Gustafson, R. W. Fuller, J. H. Cardellina,
J. B. McMahon, M. J. Currens, R. W. Buckheit, S. H. Hughes,
G. M. Cragg and M. R. Boyd, J. Med. Chem., 1992, 35, 2735–2743.
111 M. J. Currens, R. J. Gulakowski, J. M. Mariner, R. A. Moran,
R. W. Buckheit, K. R. Gustafson, J. B. McMahon and
M. R. Boyd, J. Pharmacol. Exp. Ther., 1996, 279, 645–651.
112 M. J. Currens, J. M. Mariner, J. B. McMahon and M. R. Boyd,
J. Pharmacol. Exp. Ther., 1996, 279, 652–661.
113 Y. Quan, D. Motakis, R. Buckheit, Z. Q. Xu, M. T. Flavin,
M. A. Parniak and M. A. Wainberg, Antiviral Ther., 1999, 4, 203–
209.
114 Z. Q. Xu, M. G. Hollingshead, S. Borgel, C. Elder, A. Khilevich and
M. T. Flavin, Bioorg. Med. Chem. Lett., 1999, 9, 133–138.
115 R. A. Newman, W. Chen and T. L. Madden, J. Pharm. Sci., 1998,
87, 1077–1080.
116 T. Creagh, J. L. Ruckle, D. T. Tolbert, J. Giltner, D. A. Eiznhamer,
B. Dutta, M. T. Flavin and Z. Q. Xu, Antimicrob. Agents
Chemother., 2001, 45, 1379–1386.
117 D. A. Eiznhamer, T. Creagh, J. L. Ruckle, D. T. Tolbert, J. Giltner,
B. Dutta, M. T. Flavin, T. Jenta and Z. Q. Xu, HIV Clin. Trials,
2002, 3, 435–450.
118 Anonymous, The safety and effectiveness of (+)-calanolide A in HIV-
infected patients who have never taken anti-HIV drugs, http://
clinicaltrial.gov/ct2/show/NCT00005120?term¼calanolide&rank ¼
1, Accessed 1 August, 2010.
119 Y. Kashiwada, F. Hashimoto, L. M. Cosentino, C. H. Chen,
P. E. Garrett and K. H. Lee, J. Med. Chem., 1996, 39, 1016–1017.
120 D. Yu, S. L. Morris-Natschke and K. H. Lee, Med. Res. Rev., 2007,
27, 108–132.
121 F. Li, R. Goila-Gaur, K. Salzwedel, N. R. Kilgore, M. Reddick,
C. Matallana, A. Castillo, D. Zoumplis, D. E. Martin,
J. M. Orenstein, G. P. Allaway, E. O. Freed and C. T. Wild, Proc.
Natl. Acad. Sci. U. S. A., 2003, 100, 13555–13560.
122 D. E. Martin, R. Blum, J. Wilton, J. Doto, H. Galbraith,
G. L. Burgess, P. C. Smith and C. Ballow, Antimicrob. Agents
Chemother., 2007, 51, 3063–3066.
123 P. F. Smith, A. Ogundele, A. Forrest, J. Wilton, K. Salzwedel,
J. Doto, G. P. Allaway and D. E. Martin, Antimicrob. Agents
Chemother., 2007, 51, 3574–3581.
124 D. E. Martin, K. Salzwedel and G. P. Allaway, Antiviral Chem.
Chemother., 2008, 19, 107–113.
125 N. Kilgore, M. Reddick, M. Zuiderhof, D. Stanley, T. Nitz,
P. Bullock, G. Allaway and D. Martin, Characterization of
PA1050040, a second-generation HIV-1 maturation inhibitor, http://
www.panacos.com/product_4.htm, Accessed 1 August, 2010.
126 L. Xie, Y. Takeuchi, L. M. Cosentino and K. H. Lee, J. Med. Chem.,
1999, 42, 2662–2672.
127 L. Xie, Y. Takeuchi, L. M. Cosentino and K. H. Lee, Bioorg. Med.
Chem. Lett., 1998, 8, 2151–2156.
128 H. Itokawa, S. L. Morris-Natschke, T. Akiyama and K. H. Lee,
J. Nat. Med., 2008, 62, 263–280.
129 W. E. Robinson, Jr., M. Cordeiro, S. Abdel-Malek, Q. Jia,
S. A. Chow, M. G. Reinecke and W. M. Mitchell, Mol.
Pharmacol., 1996, 50, 846–855.
130 B. Yang, Z. Meng, J. Dong, L. Yan, L. Zou, Z. Tang and G. Dou,
Drug Metab. Dispos., 2005, 33, 930–936.
131 B. Yang, Z. Y. Meng, L. P. Yan, J. X. Dong, L. B. Zou, Z. M. Tang
and G. F. Dou, J. Pharm. Biomed. Anal., 2006, 40, 417–422.
Nat. Prod. Rep., 2010, 27, 1781–1800 | 1799

132 M. Baba, R. Pauwels, J. Balzarini, J. Arnout, J. Desmyter and E. De
Clercq, Proc. Natl. Acad. Sci. U. S. A., 1988, 85, 6132–6136.
133 M. Baba, R. Snoeck, R. Pauwels and E. de Clercq, Antimicrob.
Agents Chemother., 1988, 32, 1742–1745.
134 K. J. Lorentsen, C. W. Hendrix, J. M. Collins, D. M. Kornhauser,
B. G. Petty, R. W. Klecker, C. Flexner, R. H. Eckel and
P. S. Lietman, Ann. Intern. Med., 1989, 111, 561–566.
135 C. Flexner et al., Antimicrob. Agents Chemother., 1991, 35, 2544–2550.
136 B. B. Aggarwal and B. Sung, Trends Pharmacol. Sci., 2009, 30, 85–
94.
137 A. Mazumder, K. Raghavan, J. Weinstein, K. W. Kohn and
Y. Pommier, Biochem. Pharmacol., 1995, 49, 1165–1170.
Published on 25 October 2010 on http://pubs.rsc.org | doi:10.1039/C0NP00025F
Downloaded by University of Sussex on 06 August 2012
1800 | Nat. Prod. Rep., 2010, 27, 1781–1800
View Online
138 C. J. Li, L. J. Zhang, B. J. Dezube, C. S. Crumpacker and
A. B. Pardee, Proc. Natl. Acad. Sci. U. S. A., 1993, 90, 1839–1842.
139 M. R. Boyd, K. R. Gustafson, J. B. McMahon, R. H. Shoemaker,
B. R. O’Keefe, T. Mori, R. J. Gulakowski, L. Wu, M. I. Rivera,
C. M. Laurencot, M. J. Currens, J. H. Cardellina, 2nd,
R. W. Buckheit, Jr., P. L. Nara, L. K. Pannell, R. C. Sowder, 2nd
and L. E. Henderson, Antimicrob. Agents Chemother., 1997, 41,
1521–1530.
140 A. J. Bolmstedt, B. R. O’Keefe, S. R. Shenoy, J. B. McMahon and
M. R. Boyd, Mol. Pharmacol., 2001, 59, 949–954.
141 H. Zappe, M. E. Snell and M. J. Bossard, Adv. Drug Delivery Rev.,
2008, 60, 79–87.
This journal is ª The Royal Society of Chemistry 2010