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A new highly oxygenated triterpene named ganoderic acid α has been isolated from a methanol extract of the fruiting bodies of Ganoderma lucidum together with twelve known compounds. The structures of the isolated compounds were determined by spectroscopic means including 2D-NMR. Ganoderiol F and ganodermanontriol were found to be active as anti-HIV-1 agents with an inhibitory concentration of 7.8 μg ml−1 for both, and ganoderic acid B, ganoderiol B, ganoderic acid C1, 3β-5α-dihydroxy-6β-methoxyergosta-7,22-diene, ganoderic acid α, ganoderic acid H and ganoderiol A were moderately active inhibitors against HIV-1 PR with a 50% inhibitory concentration of 0.17–0.23 mM.
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ANTI-HIV-1 AND ANTI-HIV-1-PROTEASE SUBSTANCES
FROM GANODERMA LUCIDUM
SAHAR EL-MEKKAWY, MESELHY R. MESELHY, NORIO NAKAMURA, YASUHIRO TEZUKA,
MASAO HATTORI,*NOBUKO KAKIUCHI,$KUNITADA SHIMOTOHNO,$
TAKUYA KAWAHATA%and TORU OTAKE%
Research Institute for Traditional Sino-Japanese Medicines, Toyama Medical and Pharmaceutical
University, 2630 Sugitani, Toyama, 930-0194, Japan, $Institute for Virus Research, Kyoto University,
53 Kawara-machi, Showgoin, Sakyo-ku, Kyoto, 606-8397, Japan and %Osaka Prefectural Institute of
Public Health, 3-69 Nakamichi 1-chome, Higashinari-ku, Osaka, 537-0025, Japan
(Received 20 October 1997)
Key Word IndexÐGanoderma lucidum; Polyporaceae; bioactive plant products; anti-HIV-1;
HIV-1-protease; triterpene; ganoderiol F; ganodermanontriol.
AbstractÐA new highly oxygenated triterpene named ganoderic acid ahas been isolated from a methanol
extract of the fruiting bodies of Ganoderma lucidum together with twelve known compounds. The structures
of the isolated compounds were determined by spectroscopic means including 2D-NMR. Ganoderiol F and
ganodermanontriol were found to be active as anti-HIV-1 agents with an inhibitory concentration of
7.8 mgml
ÿ1
for both, and ganoderic acid B, ganoderiol B, ganoderic acid C1, 3b-5a-dihydroxy-6b-methoxy-
ergosta-7,22-diene, ganoderic acid a, ganoderic acid H and ganoderiol A were moderately active inhibitors
against HIV-1 PR with a 50% inhibitory concentration of 0.17±0.23 mM. #1998 Elsevier Science Ltd. All
rights reserved
INTRODUCTION
Over the past decade, substantial progress has been
made in de®ning strategies for the treatment of
human immunode®ciency virus (HIV)-infected dis-
ease, acquired immunode®ciency syndrome (AIDS),
where natural products can serve as a source of
structurally novel chemicals that are worth investi-
gating as speci®c inhibitors of HIV as well as its
essential enzymes, protease (PR) and reverse tran-
scriptase (RT).
The fruiting body of Ganoderma lucidum
(Japanese name: Reishi) is one of the valuable
crude drugs, which has long been used in China
and Japan as a traditional Chinese medicine or a
folk medicine for the treatment of various kinds of
diseases [1]. Several biologically active triterpenes
and sterols have been isolated from this mushroom
and proved eective as cytotoxic [2, 3], antiviral [4]
and antiin¯ammatory agents [5, 6]. Polysaccharides
and glycoproteins possessing hypoglycemic [7, 8]
and immunostimulant [9±13] activities have also
been isolated from a water extract. In the course of
our continuing search for natural products as anti-
HIV agents, a methanol extract of the fruiting
bodies was found to be moderately active for inhi-
bition of HIV-1-induced cytopathogenicity (com-
plete inhibition at 31.3 mgml
ÿ1
in MT-4 cells) and
of its essential enzyme, protease (PR) (ca. 50% inhi-
bition at 100 mgml
ÿ1
). Therefore, this extract was
selected for further fractionation. When subjected
to bioassay-guided fractionation, the extract yielded
several active compounds. This paper describes the
isolation of thirteen compounds, and their inhibi-
tory eects against HIV-1 and its enzyme PR.
RESULTS AND DISCUSSION
Isolation and structure determination of compounds
isolated from the fruiting bodies of Ganoderma luci-
dum
Bioactivity-guided fractionation of the methanol
extract enriched the anti-HIV-1 and HIV-1-PR in-
hibitory eects in two fractions B and C. Final
puri®cation of the active compounds was achieved
by repeated column chromatography and HPLC to
give thirteen compounds, 4,5,8and 9from frac-
Phytochemistry Vol. 49, No. 6, pp. 1651±1657, 1998
#1998 Elsevier Science Ltd. All rights reserved
Printed in Great Britain
0031-9422/98/$ - see front matter
PII: S0031-9422(98)00254-4
*Author to whom correspondence should be addressed.
Fax +81-764-34-5060, E-mail: saibo421@ms.toyama-
mpu.ac.jp.
1651
tion B, and 1±3,6,7and 12 from fraction C. Three
compounds 10,11 and 13 were also obtained from
fraction A. The structures of the known compounds
were identi®ed on the basis of their spectroscopic
properties when compared with those reported for
ganoderic acids A (2) [14±16], B (3) [14±16], C1
(4) [16, 17] and H (5) [18, 19], ganoderiols A (6) [20],
B(7) [20] and F (8) [21], and ganodermanontriol
(9) [20], (all were previously isolated from the same
mushroom). In addition, ergosterol (10), ergosterol
peroxide (11, previously isolated from the sponge
Ascidia nigra) [22], cerevisterol (12) [23, 24] and
3b-5a-dihydroxy-6b-methoxyergosta-7,22-diene (13)
(both were previously isolated from the mushroom
Agaricus blazei) [24] were identi®ed (Fig. 1).
Compound 1was obtained as an amorphous
powder, [a]
D
+55.58(CHCl
3
). A molecular formula
of C
32
H
46
O
9
was estimated from a molecular ion at
m/z 574 [M]
+
in its EI-MS. The UV absorption at
254 nm and the IR band at 1650 cm
ÿ1
suggested the
presence of a conjugated ketone group. The
1
H NMR spectrum of 1analyzed by the aid of
1
H
Fig. 1. Structures of compounds isolated from a methanol extract of the fruiting bodies of Ganoderma
lucidum.
S. EL-MEKKAWY et al.1652
1
H COSY and HMQC experiments showed signals
for seven methyls (including two as doublet at d
0.97 and 1.17), and three methine protons at d3.27
(dd,J= 11.0 and 4.8 Hz), 4.80 (t,J= 8.5 Hz) and
5.62 (s) (Table 1). In addition, a singlet at d2.23
for an ester methyl was also seen. The
13
C NMR
and DEPT spectra demonstrated signals character-
istic for eight methyls, seven methines (including
three oxymethines at d66.2, 77.3 and 79.1), and ele-
ven quaternary carbons (including ®ve carbonyls at
d170.2, 181.0, 193.9, 199.0 and 206.1) (Table 1).
These data suggested a highly oxygenated lanos-
tane-type triterpene close to the structures of 3,5
and ganoderic acids G (14) [18] and K (15) [25].
However, the dierence in chemical shift between
C-8 and C-9 (6.1 ppm) in 1suggested that its substi-
tution pattern in rings B and C was similar to that
of 5, when compared to that reported for 5(about
6.0 ppm) and 3/14 (about 16.5 ppm). The EI-MS
(Fig. 2a) showed prominent fragment ions at m/z
514 [M-HOAc]
+
corresponding to the loss of an
acetoxyl group (as acetic acid) from the molecule,
and successive losses of 18 mass units (m/z 496 and
478) indicating the presence of two hydroxyl
groups. The fragment ions m/z 417 [a]
+
and 115
[e]
+
(resulted from the cleavage between C-22 and
C-23) suggested the same side chain as in related
ganoderic acids. Acetylation of 1with Ac
2
O-pyri-
dine yielded a triacetate (1a) (EI-MS m/z 658 [M]
+
;
two additional ester methyl proton signals at d2.05
and 2.10), thereby providing evidence for the pre-
sence of two hydroxyl groups in 1.
The precise connectivities of 1were established
by interpretation of the HMBC data summarized in
Table 1. Long-range correlations between H-5 and
C-7/C-9; H-30 and C-8; H-19 and C-9; and H-12
and C-11 con®rmed the diketone substitution at C-
7 and C-11. Correlations between H-18 and C-12,
and between H-12 and a carbonyl carbon signal at
d170.2 (IR, 1730 cm
ÿ1
) revealed the connectivity of
the acetoxyl group at C-12. Since H-28 and H-29
were shift-correlated with C-3, a hydroxyl group
was concluded to be located at C-3. On the other
hand, the
1
H-
1
H correlations between H-15 and H-
16a and H-16b led to the presence of the other hy-
droxyl group at C-15.
The relative stereochemistry of 1was con®rmed
by measuring the NOESY and NOE dierence
spectra of 1a as shown in Fig. 2b. The NOE
observed between H-3 (d4.48, dd,J= 11.3,
4.9 Hz), H-29 (d0.91) and H-5 (d1.65), con®rmed
the b-con®guration of the acetoxyl group at C-3,
which was equatorially oriented. Similarly, the b-
con®guration of the acetoxyl group at C-12 was
inferred from the correlations observed between H-
12 (d5.67) and the proton signal at d1.42 (H-30).
Appreciable enhancement of H-15 (d5.89) upon ir-
radiation of H-30, and vice versa, together with no
evidence of spatial correlations between H-15 and
H-18 (d0.89), con®rmed the b-con®guration of an
acetoxyl group at C-15 (d68.9). The NOE between
H-17 (d2.54) and both of H-12 and H-30 (in the
NOESY spectrum of 1) con®rmed the con®guration
at C-17. As regards the stereochemistry at C-25, the
absolute con®guration was suggested to be R, when
compared with that of ganoderic acid H (3, given
the name ganoderic acid C by Hirotani et al. [26])
having the same side chain which was con®rmed by
X-ray analysis. On the basis of the above ®ndings,
compound 1was determined as 12b-acetoxy-
3b,15b-dihydroxy-7,11,23-trioxo-5a-lanosta-8-en-26-
oic acid and named ganoderic acid a.
Inhibitory eects of isolated compounds on HIV-1-
induced cytopathogenicity, HIV-1-protease and HIV-
1-reverse transcriptase
Investigation of anti-HIV-1 and PR-inhibitory ac-
tivities of compounds 1-13 showed that some com-
pounds had moderate inhibitory activity (Table 2).
In the primary screening test for anti-HIV-1 ac-
tivity, compounds 8and 9were found to inhibit
HIV-1 induced cytopathic eect (CPE) in MT-4
cells with a 100% inhibitory concentration (IC)
value of 7.8 mgml
ÿ1
for both compounds, and the
IC value for both was a half of the respective cyto-
toxic concentration (CC) value.
As for HIV-1 PR inhibitory eects, the activity
was determined by analyzing the hydrolysates of a
Table 1. NMR Spectral Data of Compound 1(in CDCl
3
)
Atom
13
C
1
H HMBC
1 33.1 t2.70, 1.18
2 27.2 t1.68 (2H)
3 77.3 d3.27 dd (11.0, 4.8) C-28, C-29
4 39.0 s
5 51.2 d1.56 d(14.5) C-4, C-6, C-7, C-9, C-10,
C-19, C-28, C-29
6 36.6 t2.65, 2.54 C-5, C-7, C-8, C-10
7 199.0 s
8 145.6 s
9 151.7 s
10 40.3 s
11 193.9 s
12 79.1 d5.62 sC-11, C-13, C-14, C-18,
CH
3
CO
13 47.9 s
14 58.5 s
15 66.2 d4.80 t(8.5)
16 38.0 t2.75, 1.92
17 44.6 d2.54
18 12.1 q0.81 sC-12, C-13, C-14, C-17
19 17.9 q1.33 sC-1, C-5, C-9, C-10
20 29.4 d2.24
21 21.5 q0.97 d(6.5) C-17, C-20, C-22
22 48.5 t2.46, 2.30
23 206.1 s
24 46.6 t2.40, 2.80
25 35.1 d2.91
26 181.0 s
27 17.1 q1.17 d(6.5) C-24, C-25, C-26
28 27.8 q1.02 sC-3, C-4, C-5, C-29
29 15.5 q0.88 sC-3, C-4, C-5, C-28
30 21.2 q1.72 sC-8, C-13, C-14
CH
3
CO 170.2 s
CH
3
CO 20.9 q2.23 sCH
3
CO
Anti-HIV agents from Ganoderma lucidum 1653
synthetic substrate in the presence or absence of the
isolated compounds using a HPLC method. Of the
tested compounds, 3and 7were found to be
the most active against HIV-1 PR with an IC
50
of
0.17 mM for both compounds. Other compounds
such as ganoderiol F (8), ganoderic acid C1 (4), 3b,
5a-dihydroxy-6b-methoxyergosta-7,22-diene (13),
ganoderic acid a(1), ganoderic acid H (5) and
ganoderiol A (6) inhibited the enzyme activity to a
similar extent (IC
50
=0.18±0.32 mM).
However, all compounds examined did not show
any inhibitory activity against another essential
enzyme, HIV-1-RT, at concentrations below
0.25 mM.
In the present experiment, we found that D
7(8)
,
D
9(11)
-lanostadiene-type triterpenes had relatively
strong anti-HIV-1 activity. On the other hand,
D
8(9)
-lanostene-type triterpenes and ergostane-type
compounds 10±12 had no inhibition of HIV-1-
induced cytopathic eects.
As to HIV-1-protease, we could not obtain any
conclusive ®ndings on the structure-activity re-
lationship. Lanostane-type triterpenes showed IC
50
of 0.17±0.32 mM, while ergosterol derivatives had
no inhibitory activity. However, it was reported
that synthetic cosalane and its derivatives had an
anti-HIV-1 eect as well as inhibitory eects on RT
and PR [27]. Several triterpenes have been described
Fig. 2. (a) Proposed mass fragmentation pattern of 1. (b) Stereostructure for 1and 1a as indicated by
dierence NOE and NOESY spectra.
S. EL-MEKKAWY et al.1654
as antiviral compounds. Glycyrrhizin shows some
limited activity against a whole range of viruses
including HIV-1 [28]. Salaspermic acid [29] and sub-
erol (a lanostane-type) [30] inhibit HIV-1 in H9
cells in the upper micromolar range. Bile acid de-
rivatives are slightly active at 10
ÿ4
M against HIV-1
in MT-4 cells [31]. Betulinic acid derivatives
(lupane-type) have been described as potent inhibi-
tors of the cytopathogenicity of HIV-1 in CEM 4
and MT-4 cells without aecting HIV-1 RT or PR
activity [32]. When compared with other triterpenes
reported, compounds 8and 9can be used as leads
to develop other related compounds with potential
anti-HIV-1 activity.
EXPERIMENTAL
General
Mps: uncorr. Optical rotations: in CHCl
3
soln at
20±268C. UV: in MeOH. IR: in KBr. EI-MS: ioniz-
ation voltage 70 eV.
1
H and
13
C NMR: 500 MHz
and 125 MHz, respectively.
Material
Fruiting bodies of Ganoderma lucidum (Leyss. ex
Fr.) Karst. (Polyporaceae) were obtained from Alps
Chemical Industries Co. (Takayama, Japan) and
Linzhi General Institute of Co. Ltd. (Tokyo,
Japan), and their specimens were deposited at the
Museum of Materia Medica, Toyama Medical and
Pharmaceutical University (Toyama, Japan).
Enzymes and chemicals
Recombinant HIV-1 PR (purity 96% by SDS-
PAGE) was purchased from Bachem
Feinchemikalien AG (Bubendort, Switzerland).
Recombinant HIV-1-RT was obtained from Eiken
Chemicals Co. Ltd. (Osaka, Japan). A template-pri-
mer, (rA)
n
Ç(dT)
12±18
for the RT assay was obtained
from Pharmacia (Uppsala, Sweden). [Methyl-H
3
]-
thymidine 5'-triphosphate (dTTP) (speci®c activity,
1.70 TBq mmol
ÿ1
) was obtained from Amersham-
Japan (Tokyo, Japan).
Isolation procedure
The chipped fruiting bodies of G. lucidum (5 kg)
was extracted with MeOH (3 20 l) at room temp.
to give 182 g of a solid extract. The MeOH extract
was suspended in 50% aq. MeOH (2 l), the organic
solvent was removed and ®ltered through a column
of Diaion HP-20 (2 l), washed with H
2
O and then
with MeOH. The MeOH eluate was evaporated in
vacuo to give 163 g of a solid extract. The MeOH
eluate (100 g) was chromatographed on silica gel,
and elution was started with hexane, CHCl
3
and
30% MeOH in CHCl
3
to give three fractions, fr. A
(18 g), fr. B (40 g) and fr. C (26 g). CC/silica gel
(C
6
H
6
±EtOAc, 6:4) of fr. A (3 g) followed by
MPLC/silica gel (C
6
H
6
±Me
2
CO, 7:3) aorded 10
(365 mg), 11 (62 mg) and 13 (10 mg). Repeated CC
of fr. B (5 g) using silica gel (C
6
H
6
±Me
2
CO, 7:3),
RP-2 (40% aq. MeOH), MPLC/Si gel (C
6
H
6
±
Me
2
CO, 7:3), and ®nally MPLC/silica gel (5%
MeOH in CHCl
3
) gave 4(8 mg), 5(9 mg), 8
(36 mg) and 9(16 mg). Similarly, repeated CC of fr.
C (4 g) yielded 1(40 mg), 2(43 mg), 3(47 mg), 6
(6 mg), 7(15 mg) and 12 (14 mg).
12b-Acetoxy-3b,15b-dihydroxy-7,11,23-trioxo-5a-
lanosta-8-en-26-oic acid (ganoderic acid a;1)
Amorphous powder, [a]
D
+558(CHCl
3
,c1.0). IR
KBr
max cm
ÿ1
: 3450 (OH), 1750 (carbonic acid C.O),
1730 (ester C.O), 1700 (C.O) and 1650sh (conju-
gated C.O). UV l
max
(log E) nm: 254 (3.8). EI-MS
m/z 574 [M]
+
, 532 [M ÿCH
2
CO]
+
, 514
[M ÿHOAc]
+
, 496 [M ÿHOAc ÿH
2
O]
+
, 478
Table 2. Inhibitory Activities of Compounds from Ganoderma lucidum against
Protease and Proliferation of HIV-1
HIV-1
HIV-PR
Compound IC (mg/ml) CC (mg/ml) IC
50
(mM)
Ganoderic acid a(1) NE >1000 0.19
Ganoderic acid A (2) (1000) >1000 >1.0
Ganoderic acid B (3) NE >1000 0.17
Ganoderic acid C1 (4) NE >1000 0.18
Ganoderic acid H (5) NE >1000 0.20
Ganoderiol A (6) NE >1000 0.23
Ganoderiol B (7) (7.8) 500 0.17
Ganoderiol F (8) 7.8 15.6 0.32
Ganodermanontriol (9) 7.8 15.6 >1.0
Ergosterol (10) NE 1000 >1.0
Ergosterol peroxide (11) NE 15.6 >1.0
Cerevisterol (12) NE 31.3 >1.0
3b-5a-Dihydroxy-6b-methoxy ergosta-7,22-diene (13). NE 15.6 0.18
IC, the minimum concentration for complete inhibition of HIV-1 induced CEP in MT-4 cells by
microscopic observation. CC, the minimum concentration for appearance of MT-4 cell toxicity by
microscopic observation. NE, not eective. (), concentration at which weak anti-HIV-1 activity
was observed.
Anti-HIV agents from Ganoderma lucidum 1655
[M ÿHOAc ÿ2H
2
O]
+
, 417 [a]
+
, 306 [g + H]
+
and
115 [e]
+
.
3b,15b-Diacetylganoderic acid a(1a)
Amorphous powder,
1
H NMR (CDCl
3
): d0.89
(3H, s,H
3
-18), 0.91 (3H, s,H
3
-29), 0.96 (3H, d,
J= 6.5 Hz, H
3
-21), 1.00 (3H, s,H
3
-28), 1.24 (3H,
s,H
3
-19), 1.24 (3H, d,J= 7.0 Hz, H
3
-27), 1.42
(3H, s,H
3
-30), 1.65 (1H, H-5), 2.05 (3H, s,CH
3
-
CO), 2.10 (3H, s,C
H
3
-CO), 2.25 (3H, s, CH
3
-CO),
4.48 (1H, dd,J= 11.3, 4.9 Hz, H-3), 5.67 (1H, s,
H-12), 5.89 (1H, t,J= 8.4 Hz, H-15).
13
C NMR: d
68.9 (C-15), 79.9 (C-3), 80.0 (C-12), 146.1 (C-8),
154.4 (C-9), 170.0, 170.5 and 170.9 (3 acetoxyl
C1O), 179.6 (C-26), 191.9 (C-11), 207.5 (C-7) and
210.3 (C-23). EI-MS m/z 658 [M]
+
, 616
[M ÿCH
2
CO]
+
, 598 [M ÿHOAc]
+
, 538
[M ÿ2HOAc]
+
, 478 [M ÿ3HOAc]
+
, 390.
Reverse transcriptase assay
The assay was performed as previously
reported [33].
Protease assay
25 ml of HIV-1-PR assay buer (Bachem HIV-1
protease assay Kit S-100) containing 2.5 mgofa
substrate, His-Lys-Ala-Arg-Val-Leu-(pNO
2
-Phe)-
Glu-Ala-NLe-Ser-NH
2
, were mixed with 2.5 mlofa
DMSO soln of test compound, then 6.25 mlof
recHIV-1-PR (0.25 mg protein) was added to the
mixture. The reaction mixture was incubated for
15 min at 378C and then stopped by addition of
2.5 ml of 10% TFA. The hydolysate and remained
substrate were quantitatively analyzed by HPLC
under the following conditions: injection volume,
5ml; column, RP-18 (4.6 150 mm, Merck), elution,
a linear gradient of acetonitrile (20±40%) in 0.1%
TFA; ¯ow rate, 1.0 ml min
ÿ1
; detection, 280 nm.
The hydrolysate and substrate were eluted at 9.44
and 4.35 min, respectively. The inhibitory activity
of the compound in the HIV-1-PR reaction
was calculated as follows: %inhibition = 100
(A
control
ÿA
sample
)/(A
control
); where Ais a relative
peak area of the hydrolysate. Under the conditions,
acetylpepstatin was used as a positive control, its
IC
50
being 0.30 mM.
Cells
The HTLV-I-carrying cell line MT-4 cells were
used. They were maintained at 378C under 5% CO
2
in RPMI-1640 medium (Flow Laboratories, Irvine,
Scotland), supplemented with 10% fetal calf serum
(FCS, Flow laboratories, North Ryde, Australia),
100 mgml
ÿ1
of streptomycin (Meiji Seika, Tokyo,
Japan) and 100 U ml
ÿ1
of penicillin G (Banyu
Pharmaceutical, Tokyo, Japan).
Virus
The LAV-1 strain of HIV-1 was obtained from
culture supernatant of MOLT-4 cells that had been
persistently infected with LAV-1.
Primary screening for anti-HIV-1activity
MT-4 cells were infected for 1 hr with HIV-1 at
TCID
50
of 0.001 per cell. Then, the cells were
washed and resuspended at 1 10
5
cells ml
ÿ1
in
RPMI-1640 medium. A 200 ml per well of the cell
suspension was cultured for 5 days in a 96-well cul-
ture plate containing various concentrations (12
doses, maximum 1000 mgml
ÿ1
and minimum
0.49 mgml
ÿ1
) of the isolated compounds. Control
assays were performed, without these compounds,
with HIV-1-infected and uninfected cultures. On
day 5, the IC of the test compound required to pre-
vent HIV-1-induced CPE completely was deter-
mined through an optical microscope and the cell
growth was examined to give the CC that reduces
the viability of MT-4 cells.
AcknowledgementsÐA part of this study was ®nan-
cially supported by Japan Health Science
Foundation (Tokyo, Japan) and Lingzhi General
Institute Co. (Tokyo, Japan).
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Anti-HIV agents from Ganoderma lucidum 1657
... Ganoderic acid α Anti-HIV protease (0.19 mM) G. lucidum [168] 15. 3-O-acetylganoderic acid B -G. lucidum [176] ...
... G. lucidum (fruit bodies) [168,214] 121. ...
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... Unlike plants, the use of mushrooms in traditional medicine is still unknown, yet they are very effective in the treatment of certain diseases in many regions of the world. Among the species used are species of the genus Trametes and species of the complex Ganoderma lucidum which are used in the treatment of cancer, diabetes, and AIDS (El-Mekkawy et al. 1998, Akbar & Yam 2011. Species of the G. lucidum complex are regarded as the most valuable medicinal mushrooms and used to treat various diseases (Paterson 2006, Grienke et al. 2014, Zhu et al. 2016. ...
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... Triterpenes isolated from Ganoderma species such as G. lucidum have been identified as potent anticancer and immunomodulatory agents Nonaka et al.; Cheng et al. and Watanabe et al. [58][59][60]. Some triterpenes, namely, ganoderic acid C and derivatives from G. lucidum are able to inhibit biosynthesis of cholesterol Wu et al. [56]; contribute to atherosclerosis protection Komoda et al. [61]; and also show antiviral, antibacterial activity Morigiwa and el-Mekkawy [62,63]. Different sterols of fruit bodies of Inonotus obliquus were recorded with anti-inflammatory properties Niedermeyer et al. and Park et al. [64,65]. ...
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Objective: Mushrooms have been valued as traditional sources of natural bioactive compounds for many centuries and targeted as promising therapeutic agents. The bioactive mycomolecules of mushrooms are reported to have antioxidant, antitumor, antidiabetic, anti-inflammatory, and antimicrobial activity, which are the important medicinal targets in terms of drug discovery today. Hence, an attempt was made in the present study, to evaluate the immunomodulatory and oxidative process of secondary metabolites from the milky mushroom Calocybe indica (P&C) var.APK2 using radical scavenging assays. Methods: The fruiting bodies of milky mushrooms were found to produce an array of mycomolecules such as phenols, flavonoids, alkaloids, tannins, terpenoids, steroids, and saponins in their methanolic extract which was confirmed using Fourier-transform infrared spectrophotometer (FT-IR) analysis and standard phytochemical studies; hence, chromatography fractions of these mushroom seem greatly promising biological activities including antioxidants. Results: The functional analysis of the secondary metabolites of these macrofungi was evaluated by the separation of potential fractions using preparative thin-layer chromatography (TLC) that revealed seven distinct bands with Rf values of 0.14, 0.26, 0.31, 0.42, 0.52, 0.70, and 0.82; the antioxidant activity was determined through TLC in situ bio autography. The quenching property of metabolite compound which was ranging from 19% to 77.9% and the half effective concentration values of 2,2-diphenyl-1-picrylhydrazyl and hydroxyl radical scavenging activity was recorded as 64.26 μg/ml and 54.5 μg/ml sample concentration, respectively. The active mycomolecules of C. indica from the TLC was, further, confirmed using Gas Chromatography–Mass Spectrometry studies. Conclusion: The present investigation of the study revealed that the antioxidant efficacy of edible milky mushroom may be further proceeded for in vivo studies for novel drug discovery.
... Solo en las últimas décadas este organismo se ha venido investigando de manera sistemática en la medicina occidental y oriental, lo que ha permitido encontrar gran cantidad de compuestos y metabolitos activos con efectos biológicos, los cuales son obtenidos a través del cultivo de G. lucidum. Estos compuestos son principalmente los polisacáridos β-(1-3)-glucanos y β-(1-6)-glucanos (Daniel 2004, Huie y Di 2004, Yang et al. 2010, los triterpenos como los ácidos ganodéricos A, B, T y Me (Gao et al. 2002, Hirotani et al. 1987, Hirotani et al. 1985, Tang et al. 2006, entre otros componentes, los cuales presentan alta actividad fisiológica-metabólica reflejada en una creciente evidencia científica, destacándose las siguientes propiedades: antihepatitis (Campos et al. 2006, Lin 2005, Lin et al. 2003, antiinflamatoria (Akihisa et al. 2007, Ko et al. 2008, Patocka 1999, anti-HIV (El-Mekkawy et al. 1998, Min et al. 1998, hipoglucemiante (Hikino et al. 1989), hipocolesterolémica (Berger et al. 2004, Hajjaj et al. 2005, antiandrogénica (Liu et al. 2007), hepatoprotectora (Zhang et al. 2002), actividad antitumoral y anti-angiogénesis (Cao y Lin 2004, Chen et al. 2004, Song et al. 2004, Stanley et al. 2005) e inmunomoduladora (Lin 2005, Liu et al. 2003). ...
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... Suwannarach, et al. [13] reviewed literatures on fungi as producers of protease inhibitors and bioactive compounds that can offer immunomodulatory activities as potential therapeutic agents of coronaviruses. El-Mekkawy, et al. [67] and Min et al [68] isolated and characterized some triterpenes called ganoderic acids from G. lucidum that were inhibitory to HIV-1. Protease inhibitors play an important role in viral replication by selectively binding to viral proteases and blocking proteolytic cleavage of the protein precursors that are necessary for the production of infectious viral particles [13]. ...
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Chapter
Medicinal plants are always acceptable for their activity towards different disease targets as they contain various extractable chemical constituents and bioactive metabolites. Plant products have been showing their unexpected activities towards different point of our life since the very first day of civilization. During metabolism process, plants produce different chemical compounds including different secondary metabolites that are not involved in the direct growth or development of the plant but in protection of the plant from different outside as well as inside enemies. Those secondary metabolites such as polyphenols, alkaloids, flavonoids, tannins, sapogenins are having different biological activities towards different diseases conditions in human body. Hundreds of secondary metabolites from different plants have been tested preclinically as well as clinically and gave excellent action against different disease such as inflammation, diabetic, carcinogenesis, depression, cytotoxicity, hepatotoxicity, microbial infection including viral infections as HIV. This chapter is mainly focused on different activities of different secondary metabolites and their mechanism of actions in different stages of the life cycle of HIV and its infection in human body including some reported compounds with potent activity. Papaverine, Apocynin, Taxifolin, Mangiferin, Genomycin I including Calanolide A, Suksdorfin, Bevirimat, etc. are well-known compounds from different natural sources that showed very potent activities in different conditions of HIV infection.
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Triterpenoids, important secondary plant metabolites made up of six isoprene units, are found widely in higher plants and are studied for their structural variety and wide range of bioactivities, including antiviral, antioxidant, anticancer, and anti-inflammatory properties. Numerous studies have demonstrated that different triterpenoids have the potential to behave as potential antiviral agents. The antiviral activities of triterpenoids and their derivatives are summarized in this review, with examples of oleanane, ursane, lupane, dammarane, lanostane, and cycloartane triterpenoids. We concentrated on the tetracyclic and pentacyclic triterpenoids in particular. Furthermore, the particular viral types and possible methods, such as anti-human immunodeficiency virus (HIV), anti-influenza virus, and anti-hepatitis virus, are presented in this article. This review gives an overview and a discussion of triterpenoids as potential antiviral agents.
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The structures of eight novel triterpenoids, ganoderiol C (1), D (2), E (3), F (4), G (5), H (6), I (7), and ganolucidic acid E (8), isolated from the fruiting body of Ganoderma lucidum were determined by spectroscopic methods. In addition, the absolute configuration at C-23 of ganolucidic acid D (9) was determined from the CD spectrum of its p-imethylaminobenzoate derivative. © 1988, Japan Society for Bioscience, Biotechnology, and Agrochemistry. All rights reserved.
Article
Seven new triterpenes, ganoderic acids F, G, and H, lucidenic acids D2 and E2, compound C5’, and compound C6, were isolated from the surface part of gills of Ganoderma lucidum. The structures 3a, la, 4a, 6a, 7a, 2b, and 5b were proposed for these compounds, respectively. Detailed analyses of the proton and carbon-13 nuclear magnetic resonance (¹H- and ¹³C-NMR) spectra were performed by using two-dimensional (2-D) ¹H—¹H and ¹H-¹³C shift correlation techniques. © 1986, The Pharmaceutical Society of Japan. All rights reserved.
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Sixteen new triterpene acids isolated as the methyl esters from the gills of Ganoderma lucidum (Polyporaceae) along with five known triterpenes. The structures of seven new compounds among them, ganoderic acids C2, E, I, and K, compounds B8 and B9 and lucidenic acid F, were elucidated. Detailed analyses of their proton and carbon-13 nuclear magnetic resonance (1H- and 13C-NMR) spectra were also done by application of two-dimensional NMR techniques.
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Two new lanostane-type triterpenoids, ganoderiol A (1) and ganoderiol B (2) were isolated from the fruiting bodies of Ganoderma lucidum, together with known ganodermanontriol (3) and ganodermatriol (4). The compounds were identified as 5∝-lanosta-7, 9(ll)-dien-3β, 24, 25, 26-tetraol (1), 15a, 26, 27-trihydroxy-5α-lanosta-7, 9(11), 24-trien-3-one (2), 24, 25, 26-trihydroxy-5α-lanosta7, 9(1l)-dien-3-one (3) and 5α-lanosta-7, 9(11), 24-trien-3β, 26, 27-triol (4), respectively.
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New highly oxidized lanostane-type triterpenoids, ganoderic acid D, E, F, and H and lucidenic acid D, E, and F, were isolated from the gills of Ganoderma lucidum and their structures were elucidated on the basis of spectral evidence.
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Four new highly oxidized lanostane-type triterpenoids, ganoderic acid G and I and ganolucidic acid A and B, were isolated from the fungus ganoderma lucidum and their structures were elucidated on the basis of spectral evidence.
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Ganoderic acid C, a new lanostane-type triterpenoid was isolated from the fruit body of Ganoderma lucidum. The structure of ganoderic acid C was elucidated by spectroscopic data and X-ray analysis of methyl ganoderate C acetate.