Absolute stereostructure of Andirolides A-G from the flower of Carapa guianensis (Meliaceae)

Article abstract of DOI:10.1016/j.tet.2010.11.028

A new gedunin, three new mexicanolides and three new phragmalin-type limonoids named Andirolides A (1), B (2), C (3), D (4), E (5), F (6), and G (7) were isolated from oil of the flower of Carapa guianensis Aublet (Meliaceae). Their absolute stereostructu

Full text of DOI:10.1016/j.tet.2010.11.028

Tetrahedron 67 (2011) 782e792
Contents lists available at ScienceDirect
Tetrahedron
journal homepage: www.elsevier.com/locate/tet
Absolute stereostructure of Andirolides AeG from the flower of Carapa guianensis
(Meliaceae)
,
Yuji Tanaka a, Takeshi Yamada a, Yasuko In a, Osamu Muraoka b, Tetsuya Kajimoto a, Reiko Tanaka a
*
a Osaka University of Pharmaceutical Sciences, 4-20-1 Nasahara, Takatsuki, Osaka 569-1094, Japan
b Faculty of Pharmaceutical Sciences, Kinki University, 3-4-1 Kowakae, Higashiosaka, Osaka 577-8502, Japan
a r t i c l e i n f o
a b s t r a c t
Article history:
Received 30 August 2010
Received in revised form 5 November 2010
Accepted 5 November 2010
Available online 13 November 2010
A new gedunin, three new mexicanolides and three new phragmalin-type limonoids named Andirolides
A (1), B (2), C (3), D (4), E (5), F (6), and G (7) were isolated from oil of the flower of Carapa guianensis
Aublet (Meliaceae). Their absolute stereostructures were determined by 2D NMR and CD spectra, and
single-crystal X-ray analysis, and all compounds were confirmed to have the C-17bH configuration.
Considering the similarity in CD spectra between Andirolide G (7) and the xyloccensins reported by Wu,
we concluded that the structures of xyloccensins should be revised so as to have the absolute config-
uration of 17R.
Keywords:
Carapa guianensis
Limonoid
Ó 2010 Elsevier Ltd. All rights reserved.
Andirolides AeG
CD spectrum
X-ray analysis
1. Introduction
on bioactive limonoids from Maliaceae plants. As expected, three
new mexicanolide and three new phragmalin-type of limonoids
Carapa guianensis AUBLET (Meliaceae), referred to andiroba or
Brazilian mahogany in Brazil and Colombia, is a towering tree,
which grows up to 40 m high in the tropical rainforests of South
America. The tree has been used not only as timber but also as
material for herbal medicines by indigenous people in the Amazon.
In particular, the oil collected from the seeds is used as an insect
repellant and as a standing medicine for the treatment of wounds
and bruises. Interestingly, gedunin-type limonoids bearing the
4,4,8-trimethyl-17-furanylsteroid (tetra-nor-triterpene) skeleton,
named Andiloride B (2), C (3), D (4), E (5), F (6), and G (7), re-
spectively, were obtained as well as a novel gedunin-type Andi-
loride A (1). In the present paper, we report the details of their
isolation, structure determination, and cytotoxic activities against
P388, KB, L1210, and HL-60 cell lines. Moreover, we discuss the
absolute structures of these compounds and propose the necessity
of reexaming those of xyloccensins QeV in the literature.7,8
2. Results and discussion
such as andirobin,1 7-deacetoxy-7-oxogedunin,2 11
ygedunin and 6 ,11 -acetoxy-epoxyazadir-
-diacetoxygedunin,3
adione, -acetoxygedunin and
-hydroxygedunin,4
,11 -diacetoxygedunin and 6
-acetoxygedunin,5 and 1,2-dihy-
dro-3
-hydroxy-7-deacetoxy-7-oxogedunin,6 were isolated from
b-acetox-
a
b
6a
Flower oil of C. guianensis was dissolved in CHCl3, the extract of
which was separated by silica gel column chromatography, me-
dium-pressure liquid chromatography (MPLC), and reverse phased
HPLC to obtain seven new limonoids (1e7) along with the known 7-
6
a
6a
6b
b
a
b
the seeds or wood; however, the presence of mexicanolide- or
phragmalin-type limonoids, which are biosynthesized via gedunin-
type limonoids, have not been reported to date. As our initial hy-
pothesis, the flower moiety, the most highly differentiated organ in
the plant body, could contain novel metabolites produced down-
stream of the biosynthetic pathways. Thus, we investigated the
components of the flower oil of C. guianensis as a part of our study
deacetoxy-7-oxogedunin (8)9 and 6 -acetoxygedunin (9).4,10
a
Andirolide A (1) was isolated as colorless needles and demon-
strated to have the molecular formula C30H36O9 ([M]þþH; m/z
541.2442, calcd for 541.2437) by HRFABMS. The 1H and 13C NMR
spectra, which were characteristic of a gedunin limonoid such as
6a
-acetoxygedunin (9),4,10 indicated the presence of five tertiary
methyls [dH 1.16, 1.20, 1.24, 1.27, and 1.47], two acetyl methyls [dH
2.02, 2.07], two methylenes, four sp3 methines including two
oxymethines [dH 5.47 (dd), 5.50 (d)], an
5.94 (d), 7.06 (d), dC 204.0 (s)], a -substituted furan ring [dH 6.47
(dd), 7.43 (t), 7.57 (dd)], five sp3 quaternary carbons including an
a,b-unsaturated ketone [dH
b
* Corresponding author. E-mail address: tanakar@gly.oups.ac.jp (R. Tanaka).
0040-4020/$ e see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.tet.2010.11.028

Y. Tanaka et al. / Tetrahedron 67 (2011) 782e792
783
acetal or hemiacetal carbon [dC 104.0 (s)], and an
a
,
b
-unsaturated
determined by HRFABMS. The 1H and 13C NMR spectra, which were
characteristic of mexicanolide,11e13 indicated the presence of four
tertiary methyls [dH 0.74, 0.90, 1.18, 1.32], two acetyl methyls (dH
2.18, 2.22), a methyl ester group (dH 3.67), four methylenes, four sp3
methines including two oxymethines [dH 5.24 (s), 5.32 (s)], five sp3
quaternary carbons including two oxycarbons [dC 72.1 (s), 83.5 (s)],
four sp2 methines including a furan ring [dH 6.50 (dd), 7.44 (t), 7.49
(br s)], and seven sp2 quaternary carbons including two acetyl C]O
lactone [dC 111.3 (d), 163.2 (s), 169.3 (s)] (Table 1). The IR spectrum
showed the presence of a hydroxyl group at nmax 3448 cmꢀ1. In
addition, a combination of the IR absorption at nmax 1672 cmꢀ1 and
the UV absorption lmax at 236 nm (log
3
3.49) supported the
Table 1
1H NMR and 13C NMR spectroscopic data of compound 1
[dC 169.8 (s), 171.2 (s)], a COOCH3 [dC 173.3 (s)], and an a,b-un-
Position
1
saturated d-lactone [dC 165.3 (s)] (Table 2). The IR absorption at nmax
1Ha (J, Hz)
13Cb
3448 cmꢀ1 showed the presence of a hydroxyl group. The combi-
1
2
3
4
5
6
7
8
7.06
5.94
d 10.2 (2)
d 10.2 (1)
155.8
126.7
204.0
44.8
nation of IR absorption at nmax 1672 cmꢀ1 and UV absorption lmax at
236 nm (log
3 3.49) supported the presence of a,b-unsaturated
carbonyl groups. After assignments of HMQC, 1He1H COSY and
HMBC spectra (Fig. 4), it became clear that 2 was a common
mexicanolide.11e13 In the NOESY spectrum, significant NOEs were
2.51
5.47
5.50
d 12.2 (6)
dd 12.2 (5) 2.4 (7)
d 2.4 (6)
47.5
69.6
73.1
45.0
36.2
41.2
15.3
observed between H-5
H-12 ; between H-11
between H-15 and H-30
from the NOESY spectrum (Fig. 5).
b
a
and H-11
and Me-19; between H-12
. The relative structure was confirmed
b
and H-30
b
; between H-9
a and
9
10
11
2.24
dd 11.7 (11b), 8.6 (11a)
a
b
and H-17b;
a
a
b
a
b
1.99
1.83
1.60
2.28
m
m
m
m
The molecular formula of Andirolide C (3) and Andirolide D (4)
was assigned as C33H42O11 ([M]þþH; m/z 615.2800, calcd for
615.2805) and C34H42O11 ([M]þþH; m/z 627.2798, calcd for
627.2805), respectively, based on HRFABMS. The UV and IR spectra
of 3 and 4 showed the presence of a hydroxyl (nmax 3445 cmꢀ1 in 3;
12
23.4
13
14
15
16
17
18
19
20
21
22
23
28
29
30
60
42.1
169.3
111.3
163.2
104.0
23.7
5.67
s
nmax 3439 cmꢀ1 in 4), an
1680 cmꢀ1
a
,
b
-unsaturated
d-lactone (nmax
1.16
1.24
s
s
;
lmax 224.6 nm in 3; lmax 227.5 nm in 4) and ester
groups (nmax 1259 cmꢀ1). According to the structures of 3 and 4,
their UV, IR, and 1H and 13C NMR spectra were very similar to those
of 2 except for the absence of an acetyl group at C-3 and the
presence of a 2-methylpropanoyl group [dH 1.25 and 1.26 (each 3H,
d, J¼6.9 Hz); 2.73 (1H, sept. J¼6.9 Hz); 5.21 (1H, s)] in 3, and the
presence of a tigloyl group [dH 1.89 (3H, d, J¼7.0 Hz), 1.94 (3H, s),
6.97 (1H, qd, J¼7.0, 1.4 Hz)] in 4 (Table 2).14 In the HMBC spectrum
of 3, cross-peaks were observed from H-3/C-30 and isopropyl
methyl/C-30, and cross-peaks were observed from H-3/C-30, tigloyl
group/C-30 in compound 4; therefore, the relative structures of 3
and 4 were established as shown in Fig. 1.
20.7
125.2
141.8
109.9
143.2
31.6
20.5
23.7
170.3
21.3
170.0
20.7
7.57
6.47
7.43
1.27
1.20
1.47
dd 2.2 (23), 1.2 (22)
dd 2.2 (23), 1.2 (21)
t 2.2 (22, 21)
s
s
s
6"
70
7"
2.07
2.02
s
s
a
Measured at 500 MHz in CDCl3.
Measured at 125 MHz in CDCl3.
b
Andirolide E (5), isolated as colorless prisms, had the molecular
formula C35H40O14 ([M]þþH; m/z 685.2493, calcd for 685.2496) as
determined by HRFABMS. The 1H and 13C NMR spectra indicated
the presence of two angular methyls [dH 0.98, 1.13 (each 3H, s)], two
acetyl methyls [dH 2.15 and 2.24 (each 3H, s)], a primary methyl
group [dH 1.05 (3H, t)], another methyl group [dH 1.69 (3H, s), dC 21.6
(q)], seven methylenes including an oxymethylene [dH 4.38 and
4.77 (each d, J¼13.8 Hz)], five sp3 methines including three oxy-
methines [dH 5.16 (s), 5.35 (s), 6.10 (s)], eight sp3 quaternary car-
bons, a furan ring [dH 6.41 (d), 7.44 (t), 7.49 (s)], and two lactones [dC
169.8 (s), 171.0 (s)] (Table 3), which were characteristic of phrag-
malin limonoids.7,8,15 Alkaline hydrolysis of 5 with KOH/MeOH
yielded a triol (5a), in which the two carbinolic methine proton
signals were considerably up-field shifted at [dH 3.61 and 4.49 (each
s)]. In the HMBC spectrum, cross-peaks were observed from Me-18/
C-12, C-13, C-14, and C-17; H-3/C-1, C-4, and C- 30; H-14/C-8, and C-
16; H-17/C-13, C-20, C-21, and C-22; H-30/C-9, C-30, and C-300
(Fig. 6). Two methyl groups at dH 2.15 and 2.24 (each 3H, s) showed
HMBC correlations with carbonyl groups dC 170.2 (s) and 169.5 (s),
respectively, indicating the presence of the two-acetyl groups. In
addition, the COSY correlation between a primary methyl [dH 1.05
(3H, t)] and methylene signals [dH 2.44 (1H, dq), 2.46 (1H, dq)],
correlated with the carbonyl carbon [dC 172.1 (s)], indicated the
presence of a propanoyl group.
presence of a,b-unsaturated carbonyl groups. Next, the results of
the 1He1H COSY analysis of 1 revealed the partial structures shown
by bold-faced lines in Fig. 2. Herein, it should be particularly noted
that H-6 (dH 5.47) was correlated with H-5 (dH 2.51) and H-7 (dH
5.50). In the HMBC spectrum, cross-peaks were observed from Me-
18/C-12, C-13, C-14, and C-17 [dC 104.0 (s)]; Me-19/C-1, C-5, C-9, and
C-10; H-5/C-4, C-6 [dC 69.6 (d)], and C-7 [dC 73.1 (d)]; H-6/C-5, C-60
[
dC 170.3 (s)], C-7 [dC 73.1 (d)], and C-8; H-15/C-13, and C-16 [dC
163.2 (s)] (Fig. 2). Furthermore, two methyl groups at dH 2.02 (3H, s)
and 2.07 (3H, s) showed HMBC correlations with carbonyl groups dC
170.0 (s) and 170.3 (s), respectively, indicating the presence of the
two-acetyl groups. Finally, the hydroxyl group was attached at C-17
to satisfy its molecular formula (C30H36O9) and the presence of
a hemiacetal carbon at dC 104.0 (s). Regarding the relative config-
uration of chiral centers, the hydroxyl group at C-17 was assigned as
b
, because NOEs were observed between Me-18 and H-9
and H-21 (Fig. 3). The relative configuration at C-6 was determined
to be because of the significant NOEs between H-6 and Me-19,
Me-29 and Me-30, and the coupling constants of H-6 [dH 5.47 (dd,
due
a, H-12a
a
J6 ¼12.2 Hz, J 6 ¼2.4 Hz)]. The configuration of C-7 was
a
b
,5
a
b,7b
to the significant NOEs between H-7 and H-15 and Me-30, and the
coupling constants of H-7 [dH 5.50 (d, J7 ¼2.4 Hz)], the same as
b,6b
9.4, 10 HMBC and NOESY spectra showing the relative structure of 1
are shown in Figs. 2 and 3.
A quaternary carbon at dC 119.4 (C-31) showing an HMBC cor-
relation with H3-32 [dH 1.69 (s)], suggested the presence of an
orthoacetyl group. A pair of geminal doublets at dH 1.73 (d,
J¼11.8 Hz) and 2.33 (d, J¼11.8 Hz) was assigned to H2-29 in A-ring.
Andirolide B (2), isolated as colorless needles, had the molecular
formula C31H38O11 ([M]þþH; m/z 587.2493, calcd for 587.2492) as

784
Y. Tanaka et al. / Tetrahedron 67 (2011) 782e792
Table 2
1H NMR and 13C NMR spectroscopic data of compounds 2e4
Position
2
3
4
1Ha (J, Hz)
13Cb
1Ha (J, Hz)
13Cb
1Ha (J, Hz)
13Cb
1
2
3
4
5
6
7
8
9
208.3
83.5
81.8 5.21
40.3
41.4 3.10 t 5.5 (6)
33.3 2.34 d 5.5 (5)
173.3
72.1
64.6 2.16 dd 13.0 (11
50.2
21.0 1.70 dtd 13.0 (11
1.42 qd 13.0 (9, 11
30.4 1.54 td 13.0 (11 , 12
1.83 ddd 13.0 (12
208.2
83.6
81.6 5.31
40.3
41.5 3.14 t 5.9 (6)
33.3 2.34 d 5.9 (5)
208.4
83.7
81.8
40.6
41.4
33.3
173.3
72.0
64.6
50.2
21.0
5.24
s
s
s
3.08 t 5.7 (6)
2.34 d 5.7 (5)
173.3
72.0
2.15 dd 13.0 (11
b
), 4.6 (11
a
)
b
), 4.6 (11
a
)
64.6 2.16 dd 13.1(11
50.3
30.3 1.70 dtd 13.1 (11
1.43 dq 13.1(9, 11
21.0 1.53 td 13.1 (11 , 12
1.83 ddd 13.1 (12
b), 4.1(11a)
10
11
12
a
1.70 dtd 13.0 (11
1.42 qd 13.0 (9, 11
1.53 td 13.0 (11 , 12
1.83 ddd 13.0 (12
b
), 4.6 (9, 12
, 12 ), 2.0 (12
), 2.0 (12
b
), 2.0 (12
a)
b
), 4.6 (9, 12
, 12 ), 2.0 (12
), 2.0 (11
b
), 2.0 (12
a
)
b
a
), 4.1 (9, 12
, 12 ), 2.2 (11
), 2.2 (11
b), 2.2 (12a)
b
a
b
a
a
b
)
a
a
b
)
a
a)
b
b
b
)
b
b
a
)
b
b
a)
30.5
a
), 4.6 (11
a
), 2.0 (11
b
)
a
), 4.6 (11
a
), 2.0 (11
b
)
a
), 4.1 (11a), 2.2 (11b)
13
14
15
16
17
18
19
20
21
22
23
28
29
30
39.5
166.8
39.6
166.8
39.6
167.0
115.6
165.1
79.1
6.26
s
115.7 6.22
165.3
79.2 5.33
21.6 1.30
17.2 1.17
120.1
141.8 7.49 br s
110.6 6.50 dd 1.8 (23), 0.7 (21)
143.0 7.44 t 1.8 (21, 22)
22.3 0.72
23.3 0.91
44.6 2.38 d 17.2 (30
4.19 d 17.2 (30
171.2
s
115.6 6.19
165.2
79.1 5.33
21.5 1.31
17.1 1.18
120.1
141.8 7.49 br s
110.6 6.50 dd 1.6 (23), 0.7 (21)
143.0 7.44 t 1.6 (21, 22)
22.4 0.73
23.3 0.93
44.7 2.42 d 17.2 (30
4.22 d 17.2 (30
171.2
21.6 2.17
175.8
34.3
s
5.32
1.32
1.18
s
s
s
s
s
s
s
s
s
21.6
17.2
120.1
141.8
110.6
143.1
23.1
7.49 br s
6.50 dd 1.8 (23), 0.9 (21)
7.44 t 1.8 (21, 22)
0.74
0.90
2.39 d 17.1 (30
4.14 d 17.1 (30
s
s
s
s
s
s
22.5
44.6
a
b
b
a
)
)
b
a
)
)
b
a
)
)
20
171.3
21.7
166.6
127.8
12.4
139.3
14.7
52.2
200
30
2.18
2.22
s
s
21.7 2.16
169.8
s
s
300
20.9 2.73 sept. 6.9 (3000 , 30000
1.26 d 6.9 (3")
)
3000
18.9 1.94
19.4 6.97 qd 7.0 (300 ) 1.4
s
000
30000
000
1.25 d 6.9 (3")
300
1.89 d 7.0 (30000
)
70
3.67
5.03
s
s
52.2 3.68
5.08
s
s
52.2 3.67
5.03
s
s
8-OH
a
Measured at 500 MHz in CDCl3.
Measured at 125 MHz in CDCl3.
b
The IR spectrum showed broad ester bands at nmax 1751, 1723, and
1243 cmꢀ1, which could be assigned to two acetyl and a propanoyl
esters. Thus, the planar structure of 5 was established as phrag-
malin-1,8,9-orthoacetate,16 and the positions of a propanoyl group,
methyls [dH 1.32, 1.48 (each 3H, s)], an acetyl methyl [dH 2.09 (3H,
s)], a propanoyl group [dH 1.16 (3H, t), 2.44 (1H, dq), 2.46 (1H, dq)],
a methyl ester group [dH 3.71 (3H, s), dC 173.8 (s)], an
a,b-un-
saturated lactone group [dH 6.62 (1H, s), dC163.5 (s)], an orthoace-
tate [dH 1.70 (3H, s), dC 16.5 (q), 119.7 (s)], a secondary hydroxyl
group [dH 3.87 (ddd)], a tertiary hydroxyl group [dC 84.1 (s)] and
a furan ring [dH 6.61 (dd), 7.53 (t), 7.64 (br s)] (Table 3). In the HMBC
spectrum, cross-peaks were observed from H-3 [dH 5.22 (s)]/C-4, C-
28, C-30 [dC 74.3 (d)], and C-30 [dC 169.1 (s)]; H-5/C-3 [dC 85.0 (s)], C-
4, C-6, C-7 [dC 173.8 (s)], C-9 [dC 86.1 (s)] and C-29; 1-OH [dH 3.40
(s)]/C-1 and C-29; H-30 [dH 5.35 (s)]/C-1, C-2, C-3 [dC 85.0 (s)], C-8
two acetyl groups, and two
d-lactones were determined from
HMBC and 1He1H COSY correlations (Fig. 6). The location of an
orthoacetate and the relative structure of 5 were determined from
the NOESY spectrum (Fig. 7) and by single-crystal X-ray diffraction
(Fig. 8). In the NOESY spectrum, significant NOEs were observed
between H-5
b
and H-12
; between H-17 and H-12
and H-12 , H-15 , and H-17
b
, H-30
b
b
and Me-28; between H-15
, H-22 and H-30 ; and between
; therefore, the C-ring adopted
b and
H-30
b
b
b
H-30
b
b
b
b
[dC 83.6 (s)], C-9, C-14 [dC 153.8 (s)], and C-31 [dC 119.7 (s)]. The
a boat-like conformation similar to switenialide D.17
positions of an acetoxyl, a propanoyl, a hydroxyl, a carbomethoxyl,
and an orthoacetyl were located by detailed 1He1H COSY and
HMBC correlations (Fig. 9). In particular, the location of the
orthoacetate was confirmed by the HMBC correlation between H-
30 and C-31, in addition to the correlation between the hydroxyl
proton at C-1 and C-29. In the NOESY spectrum, significant NOEs
Andirolide F (6), obtained from colorless needles, had the mo-
lecular formula C35H38O14 ([M]þþH; m/z 683.2335, calcd for
683.2340) as determined by HRFABMS. The UV and IR spectra
showed a,b-unsaturated d
-lactone and ester groups. The IR, 1H, and
13C NMR spectra were very similar to those of 5 except for a double
bond at C-14:15 [dH 6.05 (s), dC 121.0 (d), 159.6 (s)] (Table 3). NOESY
experiments revealed the relative stereochemistry of 6 to have the
same conformation as 5.
(Fig. 10) were observed between H-11
and Me-19; between H-12 and H-5
the C-12 hydroxyl group [dH 3.87 (1H, dd, J¼13.0 (11
was attached at the equatorial. The relative structure was estab-
a
b
and Me-18; between H-11
, H-17 and H-21, therefore,
), 4.5 (11 ))]
b
b
b
a
b
Andirolide G (7), isolated as colorless needles, had the molecular
formula C34H40O14 ([M]þþH; m/z 673.2505, calcd for 673.2495) as
determined by HRFABMS. The IR spectrum showed the presence of
a
lished as a phragmalin 8,9,30-orthoacetate analog, which was once
isolated from Xylocarpus granatum.7,8,15
a hydroxyl at nmax 3568 cmꢀ1, and ester groups at nmax 1729 cmꢀ1
.
Next, Andirolide G (7) showed two positive Cotton effects at
213 nm (D3¼þ8.5) and 264 nm (D3¼þ2.8), as did xyloccensin Q.
The 1H and 13C NMR spectra indicated the presence of two angular

Y. Tanaka et al. / Tetrahedron 67 (2011) 782e792
785
Fig. 1. Structures of compounds 1e9 isolated from C. guianensis, xyloccensins Q, and Khayanolide C.
Although both compounds should have the same absolute structure
having the configuration of C-17R, as claimed by Wu, limonoids are
known to be biosynthesized via euphane-type triterpenoid in-
termediates with 17bH; thus, it was necessary to reconfirm the
absolute configuration of xyloccensins. Herein, CD spectra of com-
pounds 1e4 and 6e7 were measured, which afforded the following
supported by the finding that compounds 6e7, which have no func-
tional groups to affect the CD spectra, showed no Cotton effects in the
region of wavelengths longer than 300 nm (Fig. 12). Moreover, X-ray
diffraction analysis of the 16-p-bromobenzoate 8d derived from the
mostabundantlimonoid,7-deacetoxyl-7-oxogedunin(8)(Scheme1),
revealed the structure to have the absolute configuration as shown in
Fig. 13. As a result, the absolute stereochemistry of Andirolides AeG
(1e7) and 8 was determined as shown in Fig. 1.
Wu may have mistaken the stereochemistry of xyloccensins be-
cause the presence of the C-2 ketonewas not considered, which could
affect the CD spectrum of Khayanolide C. In conclusion, we estimated
thattheCottoneffectscouldbepotentiallyaffectedbytheinteractions
of a chromophore and a carbonyl group. Namely, the carbonyl groups
results; compound
1
[235
(
D3¼þ9.4), 264
(
D3¼þ1.4), 299
(
(
D3¼ꢀ1.4), 340 (D3¼ꢀ2.0)]; 2 [220 (D3¼þ13.5), 261 (D3¼þ1.8), 297
D3¼ꢀ4.4)]; 3 [219 (D3¼þ11.3), 260 (D3¼þ1.6), 297 (D3¼ꢀ3.5)]; 4
[223 (D3¼þ13.7), 260 (D3¼þ1.2), 297 (De¼ꢀ3.8)]; 6 [224 (D3¼þ7.6),
270 (D3¼þ6.1)]; 7 [213 (D3¼þ8.5), 268 (D3¼þ2.8)] (Fig. 11).
Since compound 1 had an
a,b-unsaturated ketone on a six-
membered ring and compounds 2e4 had a ketone on a six-mem-
bered ring in each molecule, some interactions between the ketones
and the a,b-unsaturated lactone on C-17 would cause negative Cotton
of the
in the same direction in Khayanolide C, while the carbonyl groups of
the -unsaturated lactone and ketone at the C-1 or C-3 position in
compounds 1e4 faced in different directions (Fig. 12). Thus, the
a,b-unsaturated d-lactone and the C-2 ketone were orientated
effectsat299nm(D3¼ꢀ1.4)and340nm(D3¼ꢀ2.0)in1andat297nm
a,b
in 2e4 (D3¼ꢀ4.4,ꢀ3.5, and ꢀ3.8, respectively). This assumption was

786
Y. Tanaka et al. / Tetrahedron 67 (2011) 782e792
Fig. 2. Selected 1He1H COSY and HMBC correlations in Andirolide A (1).
Fig. 4. Selected 1He1H COSY and HMBC correlations in Andirolide B (2).
Fig. 3. Key NOESY correlations for Andirolide A (1).
negative Cotton effect at 245 nm (D3¼ꢀ 4.3) in the CD spectrum of
Khayanolide C would be caused by the interaction between parallel
orientated carbonyl groups. Moreover, the first Cotton effect for the
furan ring chromophore at 225 nm should have had a negative Cotton
effect, if xyloccensins were enantiomers.
Fig. 5. Key NOESY correlations for Andirolide B (2).
3. Experimental
Wu took the reversed sign of Dd values in applying the Mosher
method using (R)-(ꢀ)- and (S)-(þ)-MTPACl to determine the ab-
solute structure of xyloccensin Q, i.e., (R)-MTPACl should afford (S)-
esters; however, it was stated that the (R)-ester was generated.18,19
The isolated compounds 1e7 were subjected to assays of growth
inhibition using various cancer cell lines. As a primary screening for
antitumor activities, the cell growth inhibitory properties of com-
pounds 1e7 were examined using the murine P388 leukemia cell
line, the human HL-60 leukemia cell line, the murine L1210 leu-
kemia cell line and the human KB epidermoid carcinoma cell line.
Compound 1 exhibited significant cytotoxic activity against all cell
lines (Table 4).20 Compound 6 also showed moderate cytotoxic
activity.
3.1. General procedures
Melting points were determined on a Yanagimoto micro-melting
point apparatus and are uncorrected. Optical rotations were mea-
sured with a JASCO DIP-1000 digital polarimeter. IR spectra were
recorded on a PerkineElmer 1720X FTIR spectrophotometer. 1H and
13C NMR spectrawere recorded on a Varian INOVA 500 spectrometer
with standard pulse sequences, operating at 500 and 125 MHz, re-
spectively. CDCl3 was used as the solvent and TMS as the internal
standard. FABMS was recorded on a JEOL JMS-700 mass spectrom-
eter. Column chromatography (silica gel, 70e230 mesh; Merck)
and medium-pressure liquid chromatography (MPLC; silica gel,

Y. Tanaka et al. / Tetrahedron 67 (2011) 782e792
787
Table 3
1H NMR and 13C NMR data for compounds 5e7
Position
5
6
7
1Ha (J, Hz)
13Cb
85.9
1Ha (J, Hz)
13Cb
83.9
1Ha (J, Hz)
13Cb
84.1
1
2
3
4
5
6
85.2
81.3
46.2
33.0
30.8
84.6
81.3
46.4
33.7
31.4
83.4
85.0
44.6
39.9
33.7
5.16
s
5.27
s
5.22
s
2.62
2.63
2.45
dd 6.2 (6
dd 18.5 (6
dd 18.5 (6
b
), 4.0 (6
a
)
2.72
2.58
2.62
m
m
m
2.12
2.35
brd 10.5 (6)
m
a
b
b), 4.0 (5)
), 6.2 (5)
a
7
8
9
10
11
171.0
85.7
85.1
45.1
25.7
170.9
84.2
82.6
46.9
25.6
173.8
83.6
86.1
48.1
34.6
a
b
a
b
1.88
2.26
1.49
1.42
m
m
m
m
2.16
2.34
1.49
1.64
m
m
m
m
1.98
2.22
3.87
dd 17.0 (11
dd 17.0 (11
ddd 13.0 (11
b
a
), 13.0 (12)
), 4.5 (12)
12
29.1
26.5
a
), 4.5 (11
b
), 2.0 (12-OH)
66.6
13
14
15
34.4
42.8
26.5
37.6
159.6
121.0
44.8
153.8
123.7
2.04
2.72
3.22
dd 10.5 (15
dd 18.9 (15
br s 18.9 (15a)
a
), 1.6 (15
b)
a
b
b), 10.5 (14)
6.05
s
6.62
s
16
17
18
19
169.8
78.6
20.7
68.8
163.0
80.4
18.6
68.6
163.5
78.8
13.0
15.4
5.35
1.13
4.77
4.38
s
s
5.10
1.14
4.87
4.36
s
s
5.90
1.48
1.32
s
s
s
a
b
d 13.8 (19
d 13.8 (19
b
a
)
)
d 13.7 (19
d 13.7 (19
a
b
)
)
20
21
22
23
28
29
120.9
140.9
109.6
143.4
13.7
119.4
141.4
109.7
143.3
14.1
121.4
142.4
109.6
144.8
14.4
7.49
6.41
7.44
0.98
1.73
2.33
6.10
s
7.52
6.44
7.44
1.02
1.78
2.38
5.78
br s
7.64
6.61
7.53
0.74
1.72
1.96
5.35
br s
d 1.6 (23)
t 1.6 (21, 22)
s
d 11.8 (29
d 11.8 (29
s
dd 1.9 (23) 0.7 (21)
t 1.9 (21, 22)
s
d 11.4 (29
d 11.4 (29
s
dd 1.6 (23) 0.7 (21)
t 1.6 (21, 22)
pro-R
pro-S
b
a
)
)
39.0
b
a
)
)
39.2
d 10.7 (29
d 10.7 (29
s
b
a
)
)
39.8
30
31
32
20
68.5
119.4
21.6
170.2
20.9
68.1
121.0
20.9
170.0
21.8
74.3
119.7
16.5
173.9
28.1
1.69
2.15
s
s
1.68
2.17
s
s
1.70
s
200
A
B
2.44
2.46
1.16
dq 13.8 (2"B), 7.6 (2000 )
dq 13.8 (2"A), 7.6 (2000 )
t 7.6 (2")
2000
30
8.9
169.1
21.7
169.5
21.2
169.1
20.8
300
2.24
s
2.03
s
2.09
3.71
s
s
70
52.3
300
3000
172.1
27.7
173.3
27.4
A
B
2.19
2.21
1.05
dq 11.2 (3000B), 7.3 (30000)
dq 11.2 (3000A), 7.3 (30000 )
t 7.3 (3000)
2.24
2.34
1.07
m
m
30000
8.3
t 7.4 (3000)
8.5
1-OH
12-OH
3.40
1.23
s
d 2.0 (12)
a
Measured at 500 MHz in CDCl3.
Measured at 125 MHz in CDCl3.
b
230e400 mesh; Merck) were conducted. HPLC was run on a JASCO
PU-1586 instrument equipped with a differential refractometer (RI
1531). Fractions obtained from column chromatography were
monitored by TLC (silica gel 60 F254; Merck). Preparative TLC was
carried out on Merck silica gel F254 plates (20ꢁ20 cm, 0.5 mm thick).
repeatedly recrystallized from MeOH/CHCl3 to give 7-deacetoxy-7-
oxogedunin (8) (10.1 g). The yellow oil was rechromatographed over
silica gel (2 kg) giving an amorphous solid (Fr. No. 19e31, 6.0 g),
which was subjected to CC using n-hexane/AcOEt 5:1 to afford the
residue F1 (Fr. No. 43e51, 900 mg), subjected to CC with n-hexane/
AcOEt 3:1 giving the residue F2 (Fr. No. 65e76, 433 mg), subjected to
CC with n-hexane/AcOEt 2:1 affording the residues F3 (Fr. No.
80e86, 1.0 g) and F4 (Fr. No. 87e100, 900 mg), and subjected to CC
with n-hexane/AcOEt 1:1 to give F5 (Fr. No. 101e112, 380 mg). Res-
idue F1 was subjected to CC (silica gel (230e400 mesh, 100 g); n-
hexane/AcOEt 5:1) giving a crystalline solid (40 mg, Fr. No. 51e60),
which was separated by HPLC (ODS, 60% CH3CN) to afford com-
pounds 3 (12 mg) and 4 (13 mg). Residue F2 was subjected to CC
(silica gel (230e400 mesh, 100 g); n-hexane/AcOEt, 5:1e3:1) giving
an amorphous solid (93 mg, Fr. No. 19e51), which was purified by
HPLC (ODS, 60% MeOH) to afford compound 1 (15 mg). Residue F3
was subjected to CC (silica gel (230e400 mesh, 100 g); n-hexane/
AcOEt, 5:1) to give a crystalline solid (1.0 g, Fr. No. 80e86), which was
twice subjected to CC (silica gel (230e400 mesh, 100 g); n-hexane/
3.2. Materials
The oil of the flower of C. guianensis Aublet (Meliaceae), was
collected in Amazon, Brazil, in March, 2006. A voucher specimen
(CG-01-1) was deposited in the Herbarium of the Laboratory of
Medicinal Chemistry, Osaka University of Pharmaceutical Sciences.
3.3. Isolation of compounds 1e8
The flower oil of C. guianensis Aublet (Meliaceae) (500 g) was
dissolved in CHCl3, and the CHCl3 solutionwas subjected to CC (silica
gel (7 kg); CHCl3) affording a yellow oil (Fr. No. 17e21, 164.9 g), and
a crystalline solid (Fr. No. 29e30, 31.3 g). The crystalline solid was

788
Y. Tanaka et al. / Tetrahedron 67 (2011) 782e792
Fig. 6. Selected 1He1H COSY and HMBC correlations in Andirolide E (5).
Fig. 8. ORTEP drawing of Andirolide E (5).
Fig. 9. Selected 1He1H COSY and HMBC correlations in Andirolide G (7).
Fig. 7. Key NOESY correlations for Andirolide E (5).
1370, 1246, 1031, 973, 875; CD
l
nm (D3)[c 7.39ꢁ10ꢀ4 M, CH3CN]:
235 (9.4), 251 (0.7), 264 (1.4), 281 (0), 299 (ꢀ1.4), 340 (ꢀ2.0), 393
(0). 1H and 13C NMR data, see Table 1.
AcOEt, 5:1) to give a colorless solid (240 mg, Fr. No. 97e109), which
in turn was separated by HPLC (ODS, 50% CH3CN) to afford com-
pounds 2 (10 mg) and 6 (12 mg). Residue F4 was subjected to CC
(silica gel (230e400 mesh, 100 g); n-hexane/AcOEt 2:1) to give an
amorphous solid (250 mg, Fr. No. 65e72), which was purified by
HPLC (ODS, 60% MeOH) to afford compound 5 (100 mg). Residue F5
was subjected to CC (silica gel (230e400 mesh, 100 g); n-hexane/
AcOEt, 2:1) to give an amorphous solid (78 mg, Fr. No. 86e120),
which was purified by HPLC (silica gel, CHCl3/MeOH, 50:1) to afford
compound 7 (15 mg).
3.3.2. Andirolide B (2). Colorless needles; mp 138e140ꢂC (MeOH/
23
CHCl3); [
a
]
D
ꢀ16.5 (c 0.13, CHCl3); HRFABMS m/z: 587.2493
[MþH]þ (C31H39O11, calcd for 587.2492); FABMS m/z (rel int.): 587
([MþH]þ, 100), 569 (12), 527 (9), 509 (8), 467 (56), 449 (11); UV
lmax nm (log 3
): 223 (3.73); IR nmax cmꢀ1: 3448 (OH), 2926, 1734,
1680, 1458, 1375, 1240, 1030, 875; CD
l
nm (De)[c 4.37ꢁ10ꢀ4 M,
CH3CN]: 220 (13.5), 238 (0), 243 (ꢀ0.7), 249 (0), 261 (1.8), 273 (0),
297 (ꢀ4.4), 319 (0). 1H and 13C NMR data, see Table 2.
3.3.3. Andirolide C (3). Colorless needles; mp 138e140ꢂC (MeOH/
3.3.1. Andirolide A (1). Colorless needles; mp 151e153ꢂC (MeOH/
23
CHCl3); [a]
23 þ3.1 (c 0.20, CHCl3); HRFABMS m/z: 541.2442 [MþH]þ
CHCl3); [
a
]
ꢀ37.3 (c 0.08, CHCl3); HRFABMS m/z: 615.2800
D
D
[MþH]þ (C33H43O11, calcd for 615.2805); FABMS m/z (rel int.): 615
(C30H37O9, calcd for 541.2437); FABMS m/z (rel.int.):541 ([MþH]þ,
([MþH]þ, 87%), 597 (19), 555 (5), 537 (5), 527 (6), 467 (68), 449
64), 523 (36), 481 (7), 463 (6), 421 (25), 403 (7); UV lmax nm (log 3):
(17); UV nmax nm (log 3
): 224.6 (3.94); IR nmax cmꢀ1: 3445 (OH),
236 (3.49); IR nmax cmꢀ1: 3448 (OH), 2979, 1742, 1723, 1672, 1459,

Y. Tanaka et al. / Tetrahedron 67 (2011) 782e792
789
2973, 1727, 1680, 1459, 1374, 1259, 1026, 875; CD
l
nm (De)[c
4.15ꢁ10ꢀ4 M, CH3CN]: 219 (11.3), 243 (ꢀ0.2), 260 (1.6), 274 (0), 297
(ꢀ3.5), 320 (0); 1H and 13C NMR data, see Table 2.
3.3.4. Andirolide D (4). Colorless needles; mp 131e133ꢂC (MeOH/
23
CHCl3); [
a
]
ꢀ48.6 (c 0.08, CHCl3); HRFABMS m/z: 627.2798
D
[MþH]þ (C34H43O11, calcd for 627.2805); FABMS m/z (rel int.): 627
[MþH]þ, (25), 609 (5), 567 (2), 549 (3), 527 (3), 509 (1), 467 (13),
449 (4); UV nmax nm (log 3
): 227.5 (4.12); IR nmax cmꢀ1: 3439 (OH),
2952, 1727, 1680, 1459, 1374, 1259, 1027, 876; CD
l nm (De)[c
5.44ꢁ10ꢀ4 M, CH3CN]: 223 (13.7), 245 (ꢀ0.1), 260 (1.2), 272 (0), 297
(ꢀ3.8), 323 (0); 1H and 13C NMR data, see Table 2.
3.3.5. Andirolide E (5). Colorless prisms; mp 180e182ꢂC (MeOH/
23
CHCl3); [
a]
ꢀ24.3 (c 0.10, CHCl3); HRFABMS m/z: 685.2493
D
[MþH]þ (C35H41O14, calcd for 685.2496); FABMS m/z (rel int.): 685
[MþH]þ, (69), 625 (6), 551 (3), 491 (12), 449 (34); IR nmax cmꢀ1
:
2979, 1751(d-lactone), 1723, 1638, 1374, 1243, 1146, 1083, 1048,
1022, 874; 1H and 13C NMR data, see Table 2.
3.3.6. Crystal data of 5. C35H42O14, Mr 686.69, orthorhombic, space
Fig. 10. Key NOESY correlations for Andirolide G (7).
ꢀ
ꢀ
ꢀ
group: P212121, a¼9.844 (6) ꢂA, b¼15.145 (10) A, c¼21.469 (13) A,
3
ꢀ
ꢀ3
a
¼90.00ꢂ,
b
¼90.00ꢂ
g
¼90.00 , V¼3200.7 (3) A , Dx¼1.425 g/cm
,
Fig. 11. (A) CD spectrum of Andirolide A (1). (B) CD spectra of Andirolide B (2), C (3), D (4). (C) CD spectra of Andirolide F (6), G (7).
Scheme 1. Synthesis of 8d from 7-deacetoxy-7-oxogedunin (8).

790
Y. Tanaka et al. / Tetrahedron 67 (2011) 782e792
Fig. 12. The stable conformations of Andirolides A (1), B (2), and Khayanolide C calculated on Chem 3D.
Z¼4. F (000)¼1456,
m
(Mo K
a
)¼0.110 mmꢀ1, measured independent
These can be obtained free of charge at www.ccdc.cam.ac.uk/conts/
reflections 7679, reflections 7329 (I>2
s
(I)), parameters used for
retrieving.html (or from the Cambridge Crystallograpic Date Cen-
tre,12 Union Road, Cambridge CB21EZ, U.K.; fax: þ44 1223 336 033;
e-mail: deposit@ccdc.cam.ac.uk). The H-atoms were calculated
assuming idealized geometries but were not refined.
refinement 442, R1¼0.0598 (for I>2
X-ray diffraction data were collected with a Bruker AXS SMART
APEX CCD camera using graphite-monochromated Mo K radiation
s
(I)), wR2¼0.1398 (for all data).
a
(
l
¼0.71069) at 120 K for 5. The crystal structures were solved by
a direct method using the SHELXS-97 program.21 Atomic scattering
factors were taken from the International Tables for X-ray Crys-
tallography.22 Positional parameters of non-H-atoms were refined
by a full-matrix least-squares method with anisotropic thermal
parameters using the SHELXL-97 program.21 The structural data
were deposited with the following designation: 5: CCDC-773260.
3.3.7. Alkaline hydrolysis of Andirolide E (5). Compound 5 (7.6 mg)
was refluxed with a solution of 0.03 mol dmꢀ3 KOH in MeOH over
a steam bath for 6 h. Evaporation of the solvent under reduced
pressure afforded a residue, which was subjected to HPLC [ODS,
MeOH/H2O (60:40)] to afford compound 5a (4.6 mg): HRFABMS m/
z: 545.2030 [MþH]þ (C28H33O11, calcd for 545.2023); FABMS m/z

Y. Tanaka et al. / Tetrahedron 67 (2011) 782e792
791
(3.68); IR nmax cmꢀ1: 3568 (OH), 2929, 1729, 1459, 1236, 1160, 1022,
875; CD
l
nm (D3)[c 4.46ꢁ10ꢀ4 M, CH3CN]: 213 (8.5), 249 (1.8), 268
(2.8), 323 (0); 1H and 13C NMR data, see Table 3.
3.3.10. 7-Deacetoxy-7-oxogedunin (8). Colorless prisms; mp
264e266ꢂC (MeOH/CHCl3); [
a
]
23 ꢀ38.8 (c 0.178, CHCl3). Compound
D
8 was identified from published data.2
3.3.11. NABH4 reduction of 8. To a solution of 8 (104.0 mg) in MeOH
(20 ml), NaBH4 (95 mg) was added and the mixture was refluxed for
12 h. The addition of AcOEt and H2O was followed by the usual
work-up. Evaporation of the solvent under reduced pressure affor-
ded a residue (83.0 mg), which was subjected to HPLC (ODS, MeOH/
H2O, 60:40) to give 8a (55.2 mg) and 8b (19.0 mg). Compound 8a:
HRFABMS m/z: 429.2637 [MꢀOH]þ (C26H37O5, calcd for 429.2641);
FABMS m/z (rel int.): 429 ([MꢀOH], 12%), 411 (3). Compound 8b:
HRFABMS m/z: 431.2796 [MꢀOH]þ (C26H39O5, calcd for 431.2798);
FABMS m/z (rel int.): 431 ([MꢀOH], 87%), 413 (41), 395 (2); 1H NMR
d
ppm (CDCl3): 0.79 (3H, s, H3-29), 0.86 [1H, dd, J¼12.3Hz (6
2.1 Hz (6 ), H-5], 0.90 (3H, s, H3-19), 0.94 (1H, m, H-1 ), 1.00 (3H, s,
H3-28), 1.05 (3H, s, H3-18), 1.15 (3H, s, H3-30), 1.18 (1H, m, H-9), 1.20
(1H, m, H-12 ), 1.50 (1H, m, H-6 ), 1.52 (1H, m, H-11 ), 1.56 (1H, m,
H-2 ), 1.60 (1H, m, H-11 ), 1.64 (1H, m, H-1 ), 1.65 (1H, m, H-6
1.66 (1H, m, H-2 ), 1.91 (1H, m, H-12
), 3.20 [1H, dd, J¼11.4 Hz (2
4.3 Hz (2 ), 4.3 Hz (6
), H-3], 3.67 [1H, dd, J¼10.7Hz (6
[1H, dd, J¼7.8Hz (16 ), 2.8 Hz (16 ), H-15], 4.11 [1H, dd, J¼12.4Hz
(16 ), 2.8 Hz (15), H-16 ), 7.8 Hz (15),
], 4.25 [1H, dd, J¼12.4Hz (16
H-16
b),
a
a
Fig. 13. ORTEP drawing of 8d.
a
a
b
a
a
b
b
b
),
),
Table 4
Cytotoxicity of Andirolides AeG (1e7) against the P388, HL-60, L1210 and KB cell
lines
b
b
a
b
a), H-7], 3.91
b
a
Compounds
Cell line P388 Cell line HL-60 Cell line L1210 Cell line KB
IC50 (mM)a
IC50 (mM)a
IC50 (mM)a
IC50 (mM)a
b
a
a
b
], 4.99 (1H, s, H-17), 6.42 [1H, dd, J¼1.6Hz (23), 0.7 Hz (21), H-
1
2
3
4
5
6
7
3.3
19.4
>100
>100
79.9
>100
16.1
16.7
11.4
>100
>100
>100
>100
29.3
>100
>100
>100
>100
14.4
>100
>100
>100
>100
27.0
22], 7.36 [1H, t, J¼1.6Hz (22, 21)], 7.42 (1H, br s, H-21).
3.3.12. Formation of mono-p-bromobenzoate of 8b. p-Bromo-
benzoylchloride (39 mg) and DMAP (3 mg) were added to a pyri-
dine solution (2 ml) of 8b (19.0 mg), and the reaction mixture was
refluxed for 2 h. AcOEt and H2O were added and a standard work-
up followed. The solvent was evaporated under reduced pressure,
and the residue (83.0 mg) was purified by HPLC (ODS, MeOH/H2O,
80:20) to give 8c (5.6 mg).Compound 8c: HRFABMS m/z: 630.4590
[MþH]þ(C33H4479BrO7, calcd for 630.4591).
50.6
>100
0.9
>100
0.4
68.5
7.7
5-Fluoro uracilb 0.8
a
DMSO was used as vehicle.
Positive control.
b
(rel int.): 545 [MþH]þ (100), 527 (2), 509 (1), 485 (4), 425 (11); 1H
NMR
m, H-12
J¼11.0 Hz, H-29 pro-R],1.77(1H, m, H-11
(15 ), 1.4 Hz (15
), H-14], 2.18 [1H, d, J¼11.0 Hz, H-29 pro-S], 2.24
(1H, m, H-11 ), 5.1 Hz (5), H-6 ], 2.61
), 2.42 [1H, dd, J¼16.7 Hz (6
[1H, dd, J¼16.7 Hz (6 ), 4.0 Hz (5), H-6 ],2.62[1H, dd, J¼19.0 Hz
(15 ), 9.8 Hz (14), H-15 ), 4.0 Hz (6 ), H-
], 2.76 [1H, dd, J¼5.1 Hz (6
5], 2.98 (1H, s, 2-OH), 3.22 [1H, dd, J¼19.0 Hz (15 ), 1.4 Hz (14), H-
15
], 3.61 (1H, s), 3.64 [1H, d, J¼7.6 Hz (30), 30-OH], 4.36 [1H, d,
J¼13.7 Hz (19 ), H-19 ], 4.49 [1H, d, J¼7.6 Hz (30-OH), H-30], 4.76
[1H, d, J¼13.7 Hz (19 ), H-19 ], 5.45 (1H, s, H-17), 6.38 [1H, d,
d
ppm (CDCl3): 1.08 (3H, s, H3-28), 1.10 (3H, s, H3-18), 1.42 (1H,
), 1.49 (1H, m, H-12
3.3.13. Acetylation of 8c. To a solution of 8c (5.6 mg) in pyridine
(0.5 ml) was added Ac2O (1.0 ml), and the reaction mixture was left
at room temperature overnight. The mixture was concentrated dry
under reduced pressure, and the residue was purified by HPLC
(ODS, MeOH/H2O, 85:15) to give 8d (3.8 mg).
b
a
), 1.62 (3H, s, H3-32), 1.63 [1H, d,
),1.93[1H, dd, J¼9.8Hz
a
a
b
b
a
b
b
a
a
Compound 8d: Colorless prisms; mp 241e243ꢂC (MeOH/CHCl3);
23
b
b
a
a
[a]
þ4.1 (c 0.03, CHCl3); HRFABMS m/z: 715.2484 [MþH]þ
D
(C37H4879BrO9, calcdfor715.2481);FABMS m/z(relint.):715, [MþH]þ
b
(10), 657 (19) 655 (19), 639 (5), 637 (4), 455 (12) 395 (6), 377 (7). 1H
a
b
b
NMR
s, H3-28), 0.90 (3H, s, H3-19), 0.94 [1H, dd, J¼13.0Hz (6
H-5], 1.03 (1H, m, H-1 ), 1.17 (3H, s, H3-18), 1.28 (1H, m, H-12
(1H, m, 6 ), 1.52 (1H, m, H-12 ), 1.54 (1H, m, H-2 ), 1.55 (1H, m, H-
), 1.61 (1H, m, 1 ), 1.65 (1H, m, 6 ), 1.68 (1H, m, 2 ), 1.70 (1H, m,
), 2.04 (3H, s, Ac), 2.07 (3H, s, Ac), 3.59 [1H, dd, J¼10.8Hz (6 ),
), H-7], 3.92 [1H, dd, J¼8.7Hz(16 ), 2.6Hz(16 ), H-15], 4.45
[1H, dd, J¼11.6 Hz (2 ), 4.6 Hz (2 ), H-3], 4.84 [1H, dd, J¼12.3 Hz
(16 ], 4.96 [1H, dd, J¼12.3Hz (16 ), 2.6 Hz (15),
), J¼8.7 Hz (15), H-16
H-16
d
ppm (CDCl3): 0.81 (3H, s, H3-30), 0.84 (3H, s, H3-29), 0.86 (3H,
), 2.0 Hz (6 ),
), 1.42
a
b
a
J¼1.7 Hz (23), H-22], 7.36 [1H, t, J¼1.7 Hz (21, 22), H-23], 7.44 [1H, d,
a
a
J¼1.7 Hz (23), H-21].
a
b
a
11
11
a
b
b
b
b
3.3.8. Andirolide F (6). Colorless needles; mp 188e190ꢂC (MeOH/
b
23
CHCl3); [
a
]
þ15.4 (c 0.06, CHCl3); HRFABMS m/z: 683.2335
4.6Hz(6
a
b
a
D
[MþH]þ (C35H39O14, calcd for 683.2340); FABMS m/z (rel int.): 683
b
a
[MþH]þ (100), 623 (4), 507 (10), 447 (7); UV lmax nm (log
3
): 225
a
b
b
(3.94); IR nmax cmꢀ1: 3445 (OH), 2973, 1727, 1680, 1459, 1374, 1259,
a
], 6.13 (1H, s, H-17), 6.50 [1H, dd, J¼1.8Hz (23), 0.7 Hz (21), H-
1026, 875; CD
l
nm (D3)[c 3.59ꢁ10ꢀ4 M, CH3CN]: 224 (7.6), 239 (0),
22], 7.35 [1H, t, J¼1.8Hz(22, 21), H-23], 7.55 (1H, brs, H-21) 7.62 [each
244 (ꢀ1.0), 249 (0), 270 (6.1), 317 (0); 1H and 13C NMR data, see
1H, d, J¼8.7 Hz, H-2, 20], 7.98 [each 1H, d, J¼8.7 Hz, H-3, 30].
Table 3.
3.3.14. Crystal data of 8d. C37H47BrO9, Mr 715.66, monoclinic, space
3.3.9. Andirolide G (7). Colorless needles; mp 145e147ꢂC (MeOH/
group: P21, a¼7.1529 (12) A, b¼27.995 (5) A, c¼9.411 (17) A,
ꢀ
ꢀ
ꢀ
ꢀ
CHCl3); [
a
]
D
23 þ3.2 (c 0.17, CHCl3); HRFABMS m/z: 673.2505 [MþH]þ
a
¼90.00ꢂ,
b
¼112.017 (3)ꢂ
¼90.00 , V¼1747.0 (5) A , Dx¼1.360 g/
g
3
ꢂ
(C34H41O14, calcd for 673.2495); FABMS m/z (rel.int.): 673 [MþH]þ
cmꢀ3, Z¼2. F (000)¼752,
m
(Mo K
a
)¼1.230 mmꢀ1, measured in-
(17), 655 (5), 613 (6), 599 (69), 479 (7); UV lmax nm (log
3): 229
dependent reflections 6241, reflections 3635 (I>2s (I)), parameters

792
Y. Tanaka et al. / Tetrahedron 67 (2011) 782e792
used for refinement 423, R1¼0.0645 (for I>2
s
(I)), wR2¼0.1507 (for
supported by a Grant-in-Aid for High Technology from the Ministry
of Education, Sciences, Sports and Culture, Japan.
all date), Flack
c
parameter¼0.025(12). X-ray diffraction data were
collected with a Bruker AXS SMART APEX CCD camera using
graphite-monochromated Mo K radiation (
¼0.71069) at 293 K for
a
l
Supplementary data
8d. The crystal structures were solved by a direct method using the
SHELXS-97 program.16 Atomic scattering factors were taken from
the International Tables for X-ray Crystallography.17 Positional pa-
rameters of non-H-atoms were refined by a full-matrix least-
squares method with anisotropic thermal parameters using the
SHELXL-97 program.18 The structural data were deposited with the
following designation: 8d: CCDC-773261. These can be obtained
free of charge at www.ccdc.cam.ac.uk/conts/retrieving.html (or
from the Cambridge Crystallograpic Date Centre, 12 Union Road,
Cambridge CB21EZ, U.K.; fax: þ44 1223 336 033; e-mail: depos-
it@ccdc.cam.ac.uk). The H-atoms were calculated assuming ideal-
ized geometries but were not refined.
Supplementary data related to this article can be found online
version, at doi:10.1016/j.tet.2010.11.028.
References and notes
1. Ollis, W. D.; Ward, A. D.; Zelnik, R. Tetrahedron Lett. 1964, 2607e2614.
2. Ollis, W. D.; Ward, A. D.; Oliveira, H. M. D.; Zelnik, R. Tetrahedron 1970, 26,
1637e1645.
3. Connolly, J. D.; McCrindle, R.; Overton, K. H.; Feeney, J. Tetrahedron 1966, 22,
891e896.
4. Lavie, D.; Levy, E. C.; Zelnik, R. Bioorg. Chem. 1972, 2, 59e64.
5. Marcelle, G. B.; Mootoo, B. S. Phytochemistry 1975, 14, 2717e2718.
6. Ambrozin, A. R. P.; Leite, A. C.; Bueno, F. C.; Vieira, P. C.; Fernandes, J. B.; Bueno,
O. C.; Fernandes da Silva, M. F. G.; Pagnocca, F. C.; Hebling, M. J. A.; Bacci, M., Jr. J.
Braz. Chem. Soc. 2006, 17, 542e547.
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11. Okorie, D. A.; Taylor, D. A. H. Phytochemistry 1968, 7, 1683e1686.
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turforsch. 2003, 58, 1216e1219.
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1582e1585.
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2334e2339.
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Iwagawa, T. Phytochemistry 2004, 65, 2833e2841.
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2006, 69, 891e895.
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wat, S.; Larpornmatulee, N.; Thebtaranonth, Y. Phytochemistry 2004, 65,
2569e2575.
3.3.15. Cytotoxic assay against P388, HL-60, L1210, and KB cell line-
s. Cytotoxic activities of compounds 1e6 were examined by the 3-
(4,5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide
(MTT) method. P388, HL-60, L1210, and KB cells were cultured in
Eagle’s Minimum Essential Medium (10% fetal calf serum) at 37ꢂC in
5% CO2. The test material was dissolved in dimethyl sulfoxide
(DMSO) to give a concentration of 10 mM, and the solution was
diluted with Essential Medium to give concentrations of 200, 20,
and 2 mmol, respectively. Each solution was combined with each cell
suspension (1ꢁ105 cells/ml) in the medium, respectively. After in-
cubation at 37ꢂC for 72 h in 5% CO2, the grown cells were labeled
with 5 mg/ml MTT in phosphate-buffered saline (PBS), and then the
absorbance of formazan dissolved by 20% sodium dodecyl sulfate
(SDS) in 0.1 N HCl was measured at 540 nm using a microplate
reader (Model 450; BIO-RAD). Each absorbance value was expressed
as a percentage relative to the control cell suspension, which was
prepared without the test substance by the same procedure as de-
scribed above. All assays were performed three times, semi-loga-
rithmic plots were constructed from the averaged data, and the
effective dose of the substance required to inhibit cell growth by
50% (IC50) was determined.
20. Muroga, Y.; Yamada, T.; Numata, A.; Tanaka, R. Tetrahedron 2009, 65,
7580e7586.
21. Sheldrick, G. M. SHELXLS-97, Program for Structure Determination by Direct
Method of Crystal Structure; University of Gottingen: Gottingen, 1997.
22. International Tables for X-ray Crystallography; Hahn, T., Ed.; Kluwer Academic:
Dordrecht, 1996; Vol. A.
Acknowledgements
We thank Dr. Katsuhiko Minoura and Mrs. Mihoyo Fujitake (this
university) for NMR and MS measurements. This work was