Limonoids from the seeds of a hainan mangrove, xylocarpus granatum

Article abstract of DOI:10.1021/np100395w

Ten new limonoids, hainangranatumins A-J (1-10), and 25 known compounds were isolated from seeds of a Chinese mangrove, Xylocarpus granatum, collected on Hainan Island. Hainangranatumins A-E (1-5) and I and J (9 and 10) are 9,10-seco-mexicanolides, whereas hainangranatumin F (6) is a limonoid possessing an 8α,30α-epoxy ring and a C₁-C₂₉ oxygen bridge. Hainangranatumin G (7) is a limonoid with a central pyridine ring, and hainangranatumin H (8) is a phragmalin 1,8,9-ortho ester. The relative configurations of hainangranatumins A and B were established by means of single-crystal X-ray diffraction analysis, and their absolute configurations were assigned on the basis of the specific rotation of the free acids obtained from alkaline hydrolysis. This is the first report of X-ray crystallographic structures of 9,10-seco-mexicanolides with a flexible C₂-C ₃₀-C₈ linkage. Hainangranatumins I and J (9 and 10), unusual 9,10-seco-mexicanolides with a C₉-C₃₀ linkage, are proposed to be artifacts derived from hainangranatumin C and xylomexicanin A, respectively.

Full text of DOI:10.1021/np100395w

1672
J. Nat. Prod. 2010, 73, 1672–1679
Limonoids from the Seeds of a Hainan Mangrove, Xylocarpus granatum
Jian-Yu Pan,† Shi-Lin Chen,† Min-Yi Li,‡,§ Jun Li,‡,§ Mei-Hua Yang,*,† and Jun Wu*,‡
Institute of Medicinal Plant DeVelopment, Chinese Academy of Medical Sciences and Peking Union Medical College,
Beijing 100193, People’s Republic of China, Key Laboratory of Marine Bio-resources Sustainable Utilization, South China Sea Institute of
Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, People’s Republic of China, and Graduate UniVersity
of Chinese Academy of Sciences, 19 Yuquan Road, Beijing 100049, People’s Republic of China
ReceiVed June 14, 2010
Ten new limonoids, hainangranatumins A-J (1-10), and 25 known compounds were isolated from seeds of a Chinese
mangrove, Xylocarpus granatum, collected on Hainan Island. Hainangranatumins A-E (1-5) and I and J (9 and 10)
are 9,10-seco-mexicanolides, whereas hainangranatumin F (6) is a limonoid possessing an 8R,30R-epoxy ring and a
C1-C29 oxygen bridge. Hainangranatumin G (7) is a limonoid with a central pyridine ring, and hainangranatumin H (8)
is a phragmalin 1,8,9-ortho ester. The relative configurations of hainangranatumins A and B were established by means
of single-crystal X-ray diffraction analysis, and their absolute configurations were assigned on the basis of the specific
rotation of the free acids obtained from alkaline hydrolysis. This is the first report of X-ray crystallographic structures
of 9,10-seco-mexicanolides with a flexible C2-C30-C8 linkage. Hainangranatumins I and J (9 and 10), unusual 9,10-
seco-mexicanolides with a C9-C30 linkage, are proposed to be artifacts derived from hainangranatumin C and
xylomexicanin A, respectively.
Limonoids, highly oxidized triterpene derivatives from a precur-
sor with a 4,4,8-trimethyl-17-furanylsteroid skeleton, have been
found only in plants of the order Rutales. Two meliaceous
mangroves, Xylocarpus granatum and X. moluccensis, are known
for producing antifeedant limonoids, especially mexicanolides and
phragmalins. Gedunin, the first limonoid from plants of the
Xylocarpus genus, was obtained by Taylor in 1965 from the timber
of an African X. granatum. To date, more than 120 limonoids have
been isolated and identified from the plants of this genus.1-10
Extracts of X. granatum have traditionally been utilized as
astringents and emollients for the treatment of diarrhea, cholera,
and fever. The MeOH extract of its bark exhibited significant and
dose-dependent antidiarrheal activity, and the aqueous extract of
its seeds showed strong antifilarial activity against adult worms of
subperiodic Brugia malayi.11 Previously, more than 90 limonoids
have been identified from the stem bark, timber, fruits, and seeds
of X. granatum.1-10 Herein, we report the isolation and identifica-
tion of 10 new limonoids including seven 9,10-seco-mexicanolides
(1-5, 9-10), a limonoid possessing an 8R,30R-epoxy ring and a
C1-C29 oxygen bridge (6), a limonoid with a central pyridine ring
(7), and a phragmalin 1,8,9-ortho ester (8). All compounds were
isolated from the seeds of a Chinese X. granatum collected on
Hainan Island. In addition to these new compounds, 25 known ones,
6-acetylmexicanolide,12 6-acetylxylocarpin,13 6-deoxy-swietenine,14
6-hydroxymexicanolide,15 7-oxogedunin,16 angustidienolide,17 aza-
diradione,18 granaxylocarpins A and B,19 methyl 6-hydroxyango-
lensate,20 methyl angolensate,16 mexicanolide,21 piscidirol G,22
protoxylocarpins A and D,6 sapelin E acetate,23 tigloylseneganolide
A,24 xylocarpin,25 xylocarpin I,26 xyloccensins P and Q,27,28 and
xylogranatins C, D,29 N, and O,30 were also obtained. The structures
of all these compounds were established on the basis of NMR
spectroscopic and mass spectrometric data. The relative configura-
tions of compounds 1 and 2 were confirmed by means of single-
crystal X-ray diffraction analysis, and their absolute configurations
were established on the basis of the specific rotation of the free
acids obtained from alkaline hydrolysis.
Results and Discussion
Hainangranatumin A (1) was isolated as colorless crystals. The
molecular formula C32H40O10 was established by ESIMS (m/z 607.1
[M + Na]+) and NMR data, indicating that 1 had 13 degrees of
1
unsaturation. The H and 13C NMR data (Tables 1 and 2) of 1
indicated that nine of the 13 elements of unsaturation came from
twocarbonylgroups,threeesterfunctionalities,andfourcarbon-carbon
double bonds. Therefore the molecule has to be tetracyclic. The
13C NMR data with DEPT multiplicity editing experiments revealed
that 1 had seven methyl (a methoxy, a primary methyl, two
secondary methyl groups, and three tertiary methyl groups), four
methylene, 10 methine (five olefinic), and 11 quaternary carbons
(five carbonyls).
The NMR data of 1 and its 2D NMR studies (1H-1H COSY,
HSQC, HMBC, and NOESY) (Figure 1a and b) indicated the
presence of a methoxycarbonyl group [δH 3.65 s; δC 52.0 CH3;
173.4 qC], a 2-methylbutyryl group [δH 0.84 (t, J ) 7.5 Hz), 1.13
(d, J ) 7.0 Hz), 1.41 m, 1.66 m, 2.37 m; δC 11.9 CH3, 17.1 CH3,
26.1 CH2, 41.3 CH, 175.2 qC], a ꢀ-furanyl ring [δH 7.52 br s, 6.44
br s, and 7.42 br s; δC 119.6 qC, 141.4 CH, 109.8 CH, and 143.3
CH], and two carbonyl carbons [δC 208.7 qC, 198.8 qC]. An R,ꢀ-
unsaturated δ-lactone ring D, characterized by the NMR data [δH
6.14 s, 5.29 s; δC 118.8 CH, 80.2 CH, 38.4 qC, 163.3 qC, and
163.4 qC], was confirmed by HMBC correlations between H-17/
C-13, H-17/C-14, H-17/C-16, H-15/C-13, H-15/C-14, and H-15/
C-16 (Figure 1a). A proton of an olefinic methine [δH 6.94 s; δC
161.6 CH] was assigned to H-3 on the basis of its HMBC
correlations to C-1 (δC 198.8 qC), C-2 (δC 128.8 qC), C-4 (δC 36.9
qC), C-5 (δC 45.2 CH), and C-30 (δC 67.0 CH). Therefore the ∆2,3
double bond in ring A of 1 was conjugated with the C-1 carbonyl
group. Protons of a secondary methyl group [δH 1.00 (br d, J )
4.5 Hz); δC 11.5], showing HMBC correlations to C-1, C-5, and
C-10 (δC 42.8 CH), were identified as H3-19. These 1D and 2D
NMR data strongly suggested that 1 was a 9,10-seco-mexicanolide.
HMBC correlations from H2-11 [δH 2.50 (m, H-11R), 3.01 (dd, J
) 19.1, 6.1 Hz, H-11ꢀ)] to the C-9 carbonyl carbon (δC 208.7 qC)
further confirmed the 9,10-seco feature of 1.
* To whom correspondence should be addressed. Tel & Fax: (86-10)
6289-9730. E-mail: yangmeihua15@hotmail.com (M.-H.Y.). Tel: (86-20)
8445-8442. Fax: (86-20) 8445-1672. E-mail: wwujun2003@yahoo.com
(J.W.).
† Chinese Academy of Medical Sciences and Peking Union Medical
College.
‡ South China Sea Institute of Oceanology, Chinese Academy of
Sciences.
Protons of a tertiary methyl group [δH 0.95 s; δC 18.6 CH3],
showing HMBC correlations to C-12, C-13, and C-17, were
§ Graduate University of Chinese Academy of Sciences.
10.1021/np100395w  2010 American Chemical Society and American Society of Pharmacognosy
Published on Web 09/15/2010

Limonoids from Xylocarpus granatum
Journal of Natural Products, 2010, Vol. 73, No. 10 1673
Chart 1
assigned to H3-18. Protons of two tertiary methyl groups [δH 1.15 s,
δC 27.9 CH3; δH 1.09 s, δC 20.3 CH3], exhibiting HMBC
correlations to C-4, were identified as H3-28 and H3-29, respectively.
The NMR data of 1 were similar to those of granaxylocarpin B,18
except for the replacement of the 30-O-tigloyl group in granaxy-
locarpin B by a 2-methylbutyryl group. The HMBC correlation from
H-30 (δH 6.44 s) of 1 to the carbonyl carbon [δC 175.2 qC] of this
2-methylbutyryl group further confirmed the location of this acyl
function at C-30. Thus, the planar structure of 1 was identified as
a 30-O-(2-methylbutanoate) analogue of granaxylocarpin B.
The relative configuration of 1 was partially established on the
basis of NOE interactions. Significant NOE interactions (Figure
1b) between H-12ꢀ/H-17 and H-11R/H3-18 helped to establish the
R-orientation of H3-18 and ꢀ-orientation of H-17. However, the
orientation of H-5 and H-10 in ring A could not be assigned by
However, the resonances of H3-5′ [δH 1.10 (d, J ) 7.0 Hz)] of the
2-methylbutyryl group in 2 were slightly upfield shifted. This
observation disclosed the different absolute configuration at C-2
[δH 2.42 m; δC 40.9 CH ] of this group. HSQC, HMBC, and
NOESY experiments revealed that 2 was a 9,10-seco-mexicanolide
possessing the same skeleton as that of 1.
Single-crystal X-ray diffraction analysis permitted the establish-
ment of the relative configuration of 2. A computer-generated
perspective drawing of the X-ray model of 2 is given in Figure 1d.
Four rings, designated as A, C, D, and E, were found in 2, and
their conformations in the crystal were the same as those of 1.
However, the orientation of H-2 of the 2-methylbutyryl group is
opposite that of the same group in 1. Alkaline hydrolysis of 2
yielded a corresponding 2-methylbutanoic acid. The absolute
configuration of its C-2 was concluded to be S from its positive RD
value {[R] ) +10.0 (c 0.04, Me2CO)}.32,33 On the basis of the
relative configuration of 2 from single-crystal X-ray diffraction
analysis, the absolute configurations at C-5, C-8, C-10, C-13, C-17,
and C-30 in 2 were assigned as R (Figure 1d). Therefore, the full
structure of 2, named hainangranatumin B, including the absolute
configuration, was established as shown.
1
NOE interactions due to their overlapped signals in the H NMR
spectrum. Moreover, the orientation of 8-OH was difficult to
establish without essential NOE interactions.
In order to establish the relative configuration of 1, single-crystal
X-ray diffraction analysis was employed. A computer-generated
perspective drawing of the X-ray model of 1 is given in Figure 1c.
Compound 1 is a 9,10-seco-mexicanolide with ∆2,3 and ∆14,15 double
bonds. A 2-methylbutanoyloxy group is located at C-30 in an
R-orientation. The orientation of H-5, H-30, and H3-19 is ꢀ, and
that of H-10, H-2′, and 8-OH is R. Three six-membered rings in
its crystal structure, viz., A, C, and D, adopt half-chair conforma-
tions, whereas the furan ring E is planar.
Hainangranatumin C (3), colorless crystals, had the molecular
formula C30H36O10, as established by HR-TOFMS (m/z 579.2185,
calcd for [M + Na]+ 579.2206). The NMR data of 3 were similar
to those of 1 (Tables 1 and 2), except for the replacement of the
30-O-2′(R)-methylbutyryl group in 1 by a propionyl group [δH 1.11
(t, J ) 7.5 Hz), 2.35 m; δC 173.4 qC, 27.7 CH2, 9.0 CH3]. The
substructure of the propionyl group was confirmed by HMBC cross-
The RD value is negative for (R)-2-methylbutyric acid (-14.3)31
and positive for (S)-2-methylbutyric acid (+19.3, 18.9).32,33
Therefore the absolute configuration at C-2 in the 2-methylbutyryl
group of 1 can be determined according to the specific rotation of
its acid, which was obtained from the alkaline hydrolysis of 1. The
absolute configuration at C-2′ was suggested as R by the negative
1
peaks between H3-3′/C-2′, H2-2′/C-1′, and H2-2′/C-3′ and H-1H
COSY correlations between H3-3′/H2-2′. The HMBC cross-peak
from H-30 (δH 6.50 s) to the carbonyl carbon (δC 173.4) of the
propionyl group assigned its location at C-30. Thus, the structure
of 3, named hainangranatumin C, was established as shown.
Hainangranatumin D (4) was obtained as a white, amorphous
powder. The molecular formula C29H36O10 was established by HR-
TOFMS (m/z 567.2173, calcd for [M + Na]+ 567.2206; m/z
545.2371, calcd for [M + H]+ 545.2387), indicating 12 degrees of
RD value {[R]25 -6.7 (c 0.03, Me2CO)} of the 2-methylbutyric
D
acid. On the basis of the aforementioned relative configuration of
1 from single-crystal X-ray diffraction analysis, the other six
stereogenic centers at C-5, C-8, C-10, C-13, C-17, and C-30 in 1
were assigned as R (Figure 1c). Therefore, the full structure of 1,
named hainangranatumin A, including the absolute configuration,
was unambiguously established as shown.
1
unsaturation. The H and 13C NMR data (Tables 1 and 2) of 4
indicated that eight of the 12 elements of unsaturation came from
two carbonyl groups, three carbon-carbon double bonds, and three
ester functionalities. Therefore the molecule was tetracyclic. The
NMR data of 4 were similar to those of 1, except for the presence
of an acetoxy group and the absence of a ∆14,15 double bond and a
Hainangranatumin B (2), colorless crystals, had the same
molecular formula as that of 1. The NMR data of 2, indicating the
presence of a 2-methylbutyryl group, were similar to those of 1.

1674 Journal of Natural Products, 2010, Vol. 73, No. 10
Pan et al.
Table 1. 1H (500 MHz) NMR Data (δ) for Compounds 1-5 in CDCl3 (J in Hz)
position
1
2
3
4
5
3
5
6a
6b
6.94 s
2.24a
6.97 s
2.26a
6.97 s
2.26a
6.80 s
2.24a
6.94 s
2.24a
2.28 m
2.46 m
2.24a
2.24a
2.26a
2.26a
2.24a
2.48 m
2.47 m
2.44 m
2.46 m
10
2.24a
2.26a
2.26a
2.24a
11R
11ꢀ
12R
12ꢀ
14
15R
15ꢀ
17
18
19
21
22
2.50 m
3.01 dd (19.1, 6.1)
1.62 m
2.52 m
2.51 m
1.72 m
2.33 m
2.52 m
2.55 m
3.12 dd (19.8, 6.8)
1.65 m
3.04 dd (19.5, 6.5)
3.01 dd (19.7, 6.6)
1.62 dd (13.6, 7.1)
2.60 m
1.63a
2.56 m
2.60 m
2.67 ddd (18.4, 6.1, 6.1)
2.13 br d (8.0)
2.77 dd (17.5, 8.0)
3.11 dd (17.5, 7.5)
5.36 s
2.88 m
6.14 s
6.15 s
6.13 s
6.17 s
5.29 s
0.95 s
5.31 s
0.96 s
5.31 s
0.95 s
5.34/5.31 sb
1.01/0.99 sb
1.03 d (4.2)
1.06a
1.00 br d (4.5)
7.52 br s
6.44 br s
7.42 br s
1.15 s
1.02 br d (4.5)
7.54 br s
6.45 br s
7.43 br s
1.16 s
1.01 d (5.8)
7.53 br s
6.44 br s
7.42 br s
1.17 s
1.06a
7.52 s
6.45 br s
7.43/7.40 br sb
6.24 br s
1.17 s
23
28
7.43 br s
1.14 ma
29
1.09 s
1.11 s
1.09 s
1.06 s
1.11 s
30
6.44 s
6.47 s
6.50 s
6.30 s
6.42 s
7-OMe
8-OH
30-Acyl
2′
3.65 s
3.89 s
3.67 s
3.88 s
3.66 s
3.88 s
3.68 s
4.38 s
3.69 s
3.93/3.92 sb
2.37 m
1.41 m
1.66 m
0.84 t (7.5)
1.13 d (7.0)
2.42 m
2.35 m
1.11 t (7.5)
2.10 s
2.39 m
1.42 m
1.70 m
3′a
3′b
4′
1.50 m
1.63a
0.86 t (7.5)
1.10 d (7.0)
0.85 t (7.5)/0.86 t (7.5)b
1.14 d (7.5)
5′
a Overlapped signals assigned by HSQC and HMBC spectra without designating multiplicity. b Pairs of signals resulted from C-23 epimers.
Table 2. 13C (125 MHz for 1 and 2 and 100 MHz for 3-5) NMR Data (δ) for Compounds 1-5 in CDCl3
position
1
2
3
4
5
1
2
3
4
198.8 qC
128.8 qC
161.6 CH
36.9 qC
198.8 qC
128.7 qC
161.7 CH
36.9 qC
198.7 qC
128.6 qC
161.9 CH
36.8 qC
199.7 qC
131.0 qC
160.4 CH
36.7 qC
198.6 qC
128.84/128.79 qCb
161.50/161.37 CHb
36.9 qC
5
6
7
8
45.2 CH
34.6 CH2
173.4 qC
80.1 qC
208.7 qC
42.8 CH
33.0 CH2
25.7 CH2
38.4 qC
163.3 qC
118.8 CH
163.4 qC
80.2 CH
18.6 CH3
11.5 CH3
119.6 qC
141.4 CH
109.8 CH
143.3 CH
27.9 CH3
20.3 CH3
67.0 CH
52.0 CH3
45.2 CH
34.6 CH2
173.4 qC
80.1 qC
208.6 qC
42.8 CH
33.0 CH2
25.6 CH2
38.3 qC
163.3 qC
118.6 CH
163.5 qC
80.2 CH
18.6 CH3
11.5 CH3
119.6 qC
141.4 CH
109.8 CH
143.2 CH
27.9 CH3
20.3 CH3
67.0 CH
52.0 CH3
45.2 CH
34.6 CH2
173.4 qCa
80.1 qCa
208.6 qC
42.8 CH
33.0 CH2
25.5 CH2
38.3 qC
164.0 qC
118.2 CH
163.2 qC
80.1 CHa
18.5 CH3
11.6 CH3
119.6 qC
141.4 CH
109.8 CH
143.2 CH
27.8 CH3
20.5 CH3
67.1 CH
51.9 CH3
45.0 CH
34.7 CH2
173.4 qC
79.0 qC
210.7 qC
43.1 CH
29.9 CH2
33.6 CH2
36.9 qC
48.3 CH
28.7 CH2
170.9 qC
78.4 CH
22.9 CH3
11.7 CH3
120.2 qC
141.1 CH
109.8 CH
143.2 CH
27.8 CH3
20.2 CH3
70.3 CH
52.0 CH3
45.2 CH
34.7 CH2
173.4 qC
80.2 qC
208.3 qC
42.8 CH
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
28
29
30
7-OMe
30-Acyl
1′
33.0 CH2
25.9 CH2
38.57/38.47 qCb
162.46/162.37 qCb
118.5 CH
163.6 qC
78.25/78.02 CHb
b
18.57/18.49 CH3
11.6 CH3
133.21/133.13 qCb
168.85/168.56 qCb
150.17/149.52 CHb
97.19/96.50 CHb
28.0 CH3
20.4 CH3
67.5 CH
52.0 CH3
175.2 qC
41.3 CH
26.1 CH2
17.1 CH3
11.9 CH3
175.7 qC
40.9 CH
27.1 CH2
16.3 CH3
11.7 CH3
173.4 qCa
27.7 CH2
9.0 CH3
169.4 qC
21.3 CH3
175.04/174.95 qCb
41.3 CH
26.2 CH2
17.1 CH3
11.9 CH3
2′
3′
4′
5′
a Overlapped signals. b Pairs of signals resulted from C-23 epimers.
30-O-2-methylbutyryl group. It was suggested that the 30-O-2-
methylbutyryl group in 1 was replaced by a 30-acetoxy group in
4. The appearance of an aliphatic methine group [δH 2.13 (br d, J
) 8.0 Hz, H-14); δC 48.3] and an aliphatic methylene group [2.77
(dd, J ) 17.5, 8.0 Hz, H-15R), 3.11 (dd, J ) 17.5, 7.5 Hz, H-15ꢀ);
δC 28.7] and the downfield shift of C-16 (δC 170.9) in 4 suggested
the existence of the substructure CH-14-CH2-15, which was further
confirmed by HMBC cross-peaks between H2-15/C-13, H2-15/C-
14, H2-15/C-16, H-14/C-8, H-14/C-13, H-14/C-15, and H-14/C-
16. The HMBC cross-peak from H-30 (δH 6.30 s) to the carbonyl

Limonoids from Xylocarpus granatum
Journal of Natural Products, 2010, Vol. 73, No. 10 1675
Figure 1. (a) Selected 1H-1H COSY and HMBC correlations for hainangranatumin A (1); (b) key NOE correlations for 1; (c) perspective
drawing of the X-ray structure for 1; (d) perspective drawing of the X-ray structure for hainangranatumin B (2).
carbon (δC 169.4) of the acetoxy group assigned its location at C-30.
Thus, the structure of 4, named hainangranatumin D, was estab-
lished as shown.
R-orientation of H-9, H-14, and H3-18 (Figure 2b). Hence, the
structure of compound 6, named hainangranatumin F, was estab-
lished as shown.
Hainangranatumin E (5), a white, amorphous powder, had the
molecular formula C33H42O12, as established by HR-TOFMS (m/z
Hainangranatumin G (7), a colorless, amorphous powder, had
the molecular formula C28H31NO6, as established by HR-TOFMS
1
1
639.2435, calcd for [M + Na]+ 639.2417). Its H and 13C NMR
(m/z 478.2239, calcd for [M + H]+ 478.2230). The H and 13C
data (Tables 1 and 2) were similar to those of 1, except for the
replacement of the ꢀ-furanyl ring in 1 by a γ-hydroxybutenolide
group, which was characterized by pairs of protons at δH 7.43/
7.40 (H-22), 5.34/5.31 (H-17), 3.93/3.92 (8-OH), and 1.01/0.99 (H3-
18) and by pairs of carbons at δC 133.21/133.13 (C-20), 168.85/
168.56 (C-21), 150.17/149.52 (C-22), and 97.19/96.50 (C-23). The
γ-hydroxybutenolide group in 5 was the same as that in moluccensin
N5 and domesticulide.15 HMBC cross-peaks between H-17/C-20
and H-17/C-22 revealed the linkage of the γ-hydroxybutenolide
group to C-17. Moreover, carbon resonances at δ 38.57/38.47 (C-
13), 162.46/162.37 (C-14), 78.25/78.02 (C-17), 18.57/18.49 (C-
18), 128.84/128.79 (C-2), 161.50/161.37 (C-3), and 175.04/174.95
(C-1′) resonated as pairs. The appearance of pairs of the above
proton and carbon resonances in the NMR spectra of 5 suggested
the presence of C-23 epimers. Significant NOE interactions between
H-11ꢀ/H-30, H-12ꢀ/H-30, H-17/H-12ꢀ, and H-11R/H3-18 helped
to establish the ꢀ-orientation of H-17 and the R-orientation of H3-
18. Therefore, the structure of 5, named hainangranatumin E, was
established as shown.
Hainangranatumin F (6), a white, amorphous powder, had the
molecular formula C30H38O10, as established by HR-TOFMS (m/z
581.2319, calcd for [M + Na]+ 581.2363; m/z 559.2549, calcd for
[M + H]+ 559.2543). Its NMR data (Table 3) were similar to those
of xyloccensin L,34 which is a highly oxidized limonoid with an
8R,30R-epoxy ring and a C1-C29 oxygen bridge, isolated from the
same plant, except for the replacement of the 3-O-tigloyl group in
xyloccensin L by a propionyl group [δH 2.55 (q, J ) 7.5 Hz), 1.21
(t, J ) 7.5 Hz); δC 173.9, qC, 27.5 CH2, 9.2 CH3]. The strong
HMBC cross-peak from H-3 [δH 5.31, (d, J ) 10.4 Hz)] to the
carbonyl carbon (δC 173.9, qC) of the propionyl group confirmed
its location at C-3 (Figure 2a). Moreover, NOE interactions observed
in 6 from H-5 to H3-28 and H-30 concluded the ꢀ-orientation of
H-5 and H-30. Similarly, NOE interactions between H-17/H-12ꢀ
and H-17/H-15ꢀ indicated the ꢀ-orientation of H-17, and those
between H-14/H-15R, H-14/H3-18, and H-9/H3-18 established the
NMR data (Table 3) of 7 were similar to those of xylogranatin
G,30 which is a B,C-seco-limonoid with a central pyridine ring,
except for the replacement of the 3-acetoxy group in xylogranatin
G by an ethoxy group [δH 3.40 m, 1.15 (t, J ) 7.0 Hz); δC 64.5
CH2, 15.2 CH3]. HMBC correlations between H2-1′/C-2′ and H3-
2′/C-1′ confirmed the substructure of the ethoxy group, and those
between H-3/C-1′ and H2-1′/C-3 assigned its location at C-3. The
relative configuration of 7 was found to be the same as that for
xylogranatin G30 by NOE interactions. Thus, the structure of
compound 7, named hainangranatumin G, was established as shown.
Hainangranatumin H (8), a colorless, amorphous powder, had
the molecular formula C37H44O17, as established by HR-TOFMS
(m/z 783.2464, calcd for [M + Na]+ 783.2476; m/z 761.2676, calcd
1
for [M + H]+ 761.2657). Its H and 13C NMR data (Table 4)
indicated that 8 was a phragmalin ortho-ester, characterized by a
methyl singlet at δH 1.66 and its HMBC correlation to a quaternary
carbon at δC 119.2. Three oxygenated quaternary carbons at δC
85.1, 85.6, and 86.0 further supported the identification of this ortho-
ester. Moreover, the presence of four acetoxy groups [δH 1.63, δC
169.2 qC, 20.1 CH3; δH 2.19, δC 169.7 qC, 21.4 CH3; δH 2.21, δC
170.9 qC, 21.3 CH3; δH 2.09, δC 169.1 qC, 21.2 CH3] was indicated
from the NMR data of 8. HMBC correlations from H-6, H-12, and
H-30 to the carbonyl carbons of three acetoxy groups at δC 169.7
qC, 170.9 qC, and 169.1 qC, respectively, assigned their location
at C-6, C-12, and C-30 of the phragmalin nucleus (Figure 2c). A
proton at δH 2.70 (1H, br s), which was not correlated with a carbon
signal in the HSQC spectrum, indicated the presence of a hydroxy
group in 8. HMBC correlations from this proton to C-2 and C-3
suggested the location of the hydroxy group at C-2 or C-3. HMBC
correlations from H-3 to C-2 and C-4, but not from H-3 to the
carbonyl carbon at δC 169.2 qC, suggested the location of the hy-
droxy group at C-3 and the fourth acetoxy group at C-2. The upfield
shift of C-3 (δC 83.9 CH) and the downfield shift of C-2 (δC 79.7
qC) in comparison with those of xyloccensin S35 further confirmed
the above assignments. NOE interactions between H-15ꢀ/H-17,

1676 Journal of Natural Products, 2010, Vol. 73, No. 10
Pan et al.
Table 3. 1H (500 MHz) and 13C (100 MHz) NMR Data (δ) for Compounds 6 and 7 in CDCl3
6
7
position
δH (J in Hz)
δC
δH (J in Hz)
δC
1
2
3
4
5
6a
6b
97.0 qC
41.4 CH
74.7 CH
37.1 qC
35.3 CH
31.9 CH2
157.8 qC
128.5 qC
83.8 CH
36.1 qC
45.0 CH
30.6 CH2
2.99 dd (10.4, 2.3)
5.31 d (10.4)
3.82 s
2.98a
2.74 br d (10.8)
2.30 br d (16.5)
2.40 dd (16.5, 10.8)
2.55 m
2.98a
7
8
9
10
173.92 qC
61.3 qC
47.0 CH
41.4 qC
175.5 qC
123.3 qC
158.0 qC
84.1 qC
2.07 dd (13.5, 4.8)
11R
11ꢀ
12R
12ꢀ
13
1.84 m
18.5 CH2
3.12 m
3.19 m
1.87 ddd (13.0, 5.1, 1.9)
1.77 m
27.9 CH2
1.74 td (13.6, 3.2)
1.37 td (13.6, 3.2)
2.01 td (13.6, 3.2)
33.7 CH2
30.2 CH2
36.0 qC
45.7 CH
32.4 CH2
37.6 qC
157.3 qC
111.0 CH
14
1.70 br d (5.5)
2.79 dd (16.8, 5.5)
3.11 dd (16.8, 11.7)
15R
15ꢀ
16
17
18
19
20
21
22
6.54 s
171.3 qC
79.3 CH
26.0 CH3
14.8 CH3
120.4 qC
141.0 CH
110.1 CH
143.2 CH
14.9 CH3
67.5 CH2
165.1 qC
81.0 CH
15.8 CH3
28.5 CH3
119.8 qC
141.4 CH
110.0 CH
143.3 CH
24.1 CH3
20.4 CH3
5.24 s
1.04 s
1.05 s
5.23 s
1.17 s
1.76 s
7.54 s
7.55 br s
6.52 br s
7.48 br s
1.15 s
6.44 br s
7.43 br s
0.60 s
3.89 d (8.0)
3.53 d (8.0)
3.18 d (2.3)
3.70 s
23
28
29pro-S
29pro-R
30
7-OMe
1-OH
0.67 s
60.4 CH
52.2 CH3
7.79 s
133.7 CH
3.76 s
3-Propionyl
3-Ethoxy
3.40 m
1.15 t (7.0)
1′
2′
3′
173.9 qC
27.5 CH2
9.2 CH3
64.5 CH2
15.2 CH3
2.55 q (7.5)
1.21 t (7.5)
a Overlapped signals assigned by HSQC and HMBC spectra without designating multiplicity.
H-15ꢀ/H-30, H-17/H-12, and H-17/H-30 established the ꢀ-orienta-
tion of H-12, H-17, and H-30. Similarly, those between H-14/H3-
18 and H-14/H-15R indicated the R-orientation of H-14 and H3-18
(Figure 2d), and those between H-30/H-5 and H-5/H3-28 indicated
the ꢀ-orientation of H-5 and H3-28. The NOE interaction from H-3
to Hpro-R-29, but not from H-3 to H-5, H3-28, and H-30, suggested
the R-orientation of H-3. Thus, the structure of compound 8, named
hainangranatumin H, was established as shown.
the ꢀ-orientation of H-17 and H-30. Similarly, those between H-5/
H3-19 and H-5/H3-28 proved the ꢀ-orientation of H-5 and H3-19.
Thus, the structure of compound 9, named hainangranatumin I, was
established as shown.
Hainangranatumin J (10) had the molecular formula C31H38O10, as
established by HR-TOFMS (m/z 593.2352, calcd for [M + Na]+
593.2363). The NMR data of 10 were similar to those of 9 (Table 5),
except for the replacement of the 30-O-propionyl moiety in 9 by an
isobutyryl group [δH 2.46 m, 1.11 (d, J ) 7.0 Hz), 1.09 (d, J ) 7.0
Hz); δC 174.8 qC, 33.8 CH, 18.8 CH3, 18.8 CH3]. The substructure of
the isobutyryl group was confirmed by HMBC cross-peaks between
Hainangranatumin I (9) had the same molecular formula as that
of 3, viz., C30H36O10, as established by HR-TOFMS (m/z 579.2239,
calcd for [M + Na]+ 579.2206). It indicated that 9 might be an
1
H-2′/C-1′, H-2′/C-3′, and H-2′/C-4′ and H-1H COSY correlations
1
isomer of 3. Comparison of its H and 13C NMR data (Table 5)
between H-2′/H3-3′ and H-2′/H3-4′. The HMBC cross-peak from H-30
(δH 6.21, s) to the carbonyl carbon (δC 174.8) of the isobutyryl group
assigned its location at C-30. Thus, the structure of compound 10,
named hainangranatumin J, was established as shown.
with those of 3 revealed that structures of both compounds were
indeed closely related. However, the main differences occurred at
C-8 and C-9. HMBC correlations between H2-11/C-30 (δC 66.3
CH), H-15/C-8 (δC 200.8 qC), and H2-11/C-9 (δC 79.7 qC) (Figure
2e) suggested the linkage of the C-30 methine to C-9, which led to
the formation of a hydroxylated quaternary carbon at C-9 and a
conjugated carbonyl group at C-8. The above linkage pattern was
further confirmed by the remarkable upfield shifts of H2-11 [δH
1.79 (ddd, J ) 14.6, 14.5, 4.0 Hz, 11R), 1.67 (H, ddd, J ) 14.6,
3.6, 3.6 Hz, 11ꢀ) in 9; δH 2.51 (m, 11R), 3.01 (dd, J ) 19.7, 6.6
Hz, 11ꢀ) in 3] and the UV red shift of the molecule (UV λmax 240.7
nm in 9 and λmax 212.5 nm in 3).
The first limonoid with a C30-C9 linkage was xylogranatin D,
which was reported as a genuine natural product from seeds of a
Chinese mangove, X. granatum.29 However, it was later found to
be an artifact derived from xylogranatin C, a 9,10-seco-mexicanolide
with a C30-C8 linkage. Long-time storage of xylogranatin C in
MeOH could yield xylogranatin D.30 Hainangranatumins I (9) and
J (10) are the other two limonoids with a C30-C9 linkage. Similarly,
they could be proposed to be artifacts derived from hainangrana-
tumin C (3) and xylomexicanin A,8 respectively, though the latter
was not obtained in our experiment.
The relative configuration of 9 was determined by NOE in-
teractions as shown in Figure 2f. The NOE interaction between
H-12R/H3-18 indicated the R-orientation of H3-18, whereas NOE
interactions between H-30/H-17, H-30/12ꢀ, and H-30/11ꢀ indicated
To date, ten 9,10-seco-mexicanolides have been reported. They
have been identified from seeds of the Chinese mangrove, X.

Limonoids from Xylocarpus granatum
Journal of Natural Products, 2010, Vol. 73, No. 10 1677
configurations of hainangranatumins A and B were unequivocally
determined on the basis of their X-ray crystallographic structures
and the specific rotations of their 2-methylbutyryl groups. This is
the first report of X-ray crystallographic structures of 9,10-seco-
mexicanolides with a flexible C2-C30-C8 linkage. In addition,
hainangranatumin A is the first limonoid with a 2R-methylbutyryl
group from plants of the Xylocarpus genus. This study also
demonstrates that X. granatum is a rich source for the production
of novel limonoids.
Experimental Section
General Experimental Procedures. Melting points were measured
on an X4 micromelting point detector (Beijing Tech. Instrument Co.,
Ltd., China). Optical rotations were recorded on a Polaptronic HNQW5
automatic high-resolution polarimeter (Schmidt & Haensch Co., Ltd.).
UV spectra were obtained on a Beckman DU-640 UV spectrophotom-
eter, and NMR spectra were recorded in CDCl3 using a Bruker AV-
500 or AV-400 spectrometer with TMS as internal standard. HR-
TOFMS spectra were measured on a Thermo Scientific LTQ XL
spectrometer or Waters Acquity UPLC-Q-Tof Micro MS in positive
ion mode. Single-crystal X-ray diffraction analysis was performed on
a Bruker Smart 1000 CCD single-crystal diffractometer. Preparative
HPLC was carried out on ODS columns (YMC-Pack ODS-5-A, 250
× 10 mm i.d., 5 µm) with a Waters 2998 photodiode array detector.
For CC, silica gel (200-300 mesh) (Qingdao Marine Chemical Ind.
Co., Ltd.) and RP C18 gel (Cosmosil C18-PREP 140 µm, Nacalai
Tesque, Kyoto, Japan) were used.
Plant Material. Seeds of X. granatum were collected in June 2009
on Hainan Island, China. The identification of the plant was performed
by one of the authors (J.W.). A voucher sample (No. HNXG-09-3) is
maintained in the herbarium of the South China Sea Institute of
Oceanology.
Extraction and Isolation. Dried seeds (20 kg) of X. granatum were
extracted three times with 95% EtOH at room temperature. The extract
was concentrated under reduced pressure, followed by suspension in
H2O and extraction with EtOAc. The EtOAc extract was further
suspended in H2O and extracted with CHCl3. The resulting CHCl3
extract (357 g) was subjected to silica gel CC eluted using a
CHCl3-MeOH system (100:0-5:1) to yield 322 fractions. According
to the analytic results of thin-layer chromatography, fractions 27 to 62
were combined as portion A (44.5 g) and fractions 100 to 170 were
combined as portion B (93.4 g). Portion A was further purified using
RP C18 CC (MeCN-H2O, 50:50-100:0) to afford 101 fractions
(A1-A101), whereas portion B was purified using silica gel CC to
give seven fractions B1-B7. Fraction B7 was purified using RP C18
CC (MeCN-H2O, 30:70-100:0) to afford 95 subfractions (Ba1-Ba95).
Fractions A11 to A14 (1.8 g) from portion A were combined and
fractionated by RP C18 CC (Me2CO-H2O, 30:70-100:0) to afford
subfractions Aa1-Aa16. Subfractions Aa9 and Aa10 were combined
and purified by preparative HPLC (MeOH-H2O, 55:45) to give
compounds 6 (4.1 mg) and 6-hydroxymexicanolide (3.8 mg). Subfrac-
tions Aa12 and Aa13 were combined and purified by preparative HPLC
(MeCN-H2O, 49:51) to yield compounds 4 (3.8 mg), 5 (0.9 mg), 8
(2.4 mg), methyl 6-hydroxyangolensate (2.2 mg), xyloccensins P (58.2
mg) and Q (20.7 mg), and xylogranatins C (14.9 mg) and D (84.4 mg).
Fractions A26 to A35 were combined and fractionated by RP C18
CC (MeCN-H2O, 30:70-100:0) to give subfractions Ab1-Ab32.
Subfraction Ab16 was purified by preparative HPLC (MeCN-H2O,
49:51) to afford compounds 7 (1.0 mg), 7-oxogedunin (1.9 mg),
mexicanolide (12.7 mg), 6-acetylmexicanolide (6.4 mg), and 6-acetylx-
ylocarpin (1.6 mg). Subfractions Ab17 and Ab18 were combined and
purified by preparative HPLC (MeOH-H2O, 67:33) to yield compounds
3 (37.9 mg), 9 (2.1 mg), 10 (6.1 mg), angustidienolide (0.8 mg),
azadiradione (11.5 mg), methyl angolensate (44 mg), xylocarpin (3.4
mg), and xylocarpin I (3.9 mg), and subfractions Ab24-Ab26 were
combined and purified by preparative HPLC (MeOH-H2O, 51:49) to
afford 6-deoxy-swietenine (20.5 mg) and granaxylocarpin B (16.8 mg).
Fractions A46 to A49 were combined and purified by preparative HPLC
(MeCN-H2O, 53:47) to afford compounds 1 (45.1 mg), 2 (21.5 mg), and
tigloylseneganolide A (3.3 mg).
Figure 2. (a) Selected HMBC correlations for hainangranatumin
1
F (6); (b) key NOE correlations for 6; (c) selected H-1H COSY
and HMBC correlations for hainangranatumin H (8); (d) key NOE
correlations for 8; (e) selected 1H-1H COSY and HMBC correla-
tions for hainangranatumin I (9); (f) key NOE correlations for 9.
granatum,8,10,19,26,29 and can be classified into two substructural
classes, of which one possesses a 1,9-oxygen bridge to form a pyran
ring, while the other one contains a flexible C2-C30-C8 linkage.
Before this study, xylogranatin A, the structure of which was
supported by X-ray crystallographic analysis,29 has been the only
example belonging to the former class. Previously, the relative
configuration of ring A in 9,10-seco-mexicanolides of the latter
class has been derived from NOE data. However, the orientation
of H-5 and H-10 in the A ring was difficult to assign due to their
overlapped signals with one signal of H2-6. In this paper, single-
crystal X-ray diffraction analysis was successfully employed to
establish the relative configurations of two 9,10-seco-mexicanolides
with a flexible C2-C30-C8 linkage, viz., hainangranatumins A and
B (1 and 2).
In summary, seven 9,10-seco-mexicanolides, named hainan-
granatumins A-E (1-5) and I and J (9 and 10), were identified
from the seeds of a Chinese mangrove, X. granatum, collected on
Hainan Island, along with a limonoid possessing an 8R,30R-epoxy
ring and a C1-C29 oxygen bridge (6), a limonoid with a central
pyridine ring (7), and a phragmalin 1,8,9-ortho ester (8). Twenty-
five known compounds were also obtained. Hainangranatumin A
(1) was found to possess a 2R-methylbutyryl group, whereas
hainangranatumin B (2) a 2S-methylbutyryl group. They are epimers
with the same 9,10-seco-mexicanolide skeleton. The absolute
Subfractions Ba73 to BaB81 were combined and purified by
preparative HPLC (MeOH-H2O, 76:24) to give granaxylocarpin A (1.1

1678 Journal of Natural Products, 2010, Vol. 73, No. 10
Pan et al.
Table 4. 1H (500 MHz) and 13C (125 MHz) NMR Data (δ) for Compound 8 in CDCl3
position
δH (J in Hz)
δC
position
δH (J in Hz)
δC
1
2
3
4
5
6
7
8
85.1 qC
79.7 qC
83.9 CH
45.4 qC
40.2 CH
71.5 CH
169.6 qC
85.6 qC
86.0 qC
46.2 qC
31.6 CH2
22
23
28
29R
29ꢀ
30
7-OMe
3-OH
6.42 br s
7.39 br s
1.10 s
1.78 d (15.0)
2.06 d (15.0)
5.95 s
109.7 CH
143.2 CH
15.5 CH3
40.0 CH2
4.58 s
3.31 br s
6.06 br s
70.4 CH
53.0 CH3
3.81 s
2.70 br s
9
1,8,9-Trioxyethyl
10
11R
11ꢀ
12
13
14
15R
15ꢀ
16
17
18
19
20
21
1′
2′
119.2 qC
21.1 CH3
2.40 dd (14.0, 3.5)
1.81 br d (14.0)
4.61 br d (3.5)
1.66 s
1.63 s
2.19 s
2.21 s
2.09 s
2-Acetoxy
1′′
69.0 CH2
38.8 qC
43.8 CH
26.9 CH2
169.2 qC
20.1 CH3
2′′
2.24 br d (7.5)
2.78 dd (20.5, 10.5)
3.28 br d (20.5)
6-Acetoxy
1′′′
169.7 qC
21.4 CH3
2′′′
169.6 qC
77.1 CH
14.0 CH3
13.7 CH3
120.8 qC
140.9 CH
12-Acetoxy
1′′′′
5.53 s
1.22 s
1.22 s
170.9 qC
21.3 CH3
2′′′′
30-Acetoxy
1′′′′′
169.1 qC
21.2 CH3
7.46 br s
2′′′′′
Table 5. 1H (500 MHz) and 13C (100 MHz) NMR Data (δ) for Compounds 9 and 10 in CDCl3
9
10
position
δH (J in Hz)
δC
δH (J in Hz)
δC
1
2
3
4
5
6
198.6 qC
130.4 qC
161.3 CH
36.7 qC
45.4 CH
34.8 CH2
198.6 qC
130.4 qC
161.1 CH
36.7 qC
45.4 CH
34.8 CH2
6.98 s
6.94 s
2.32a
2.32a
2.32a
2.32a
2.49 m
2.50 m
7
8
9
10
11R
11ꢀ
12R
12ꢀ
13
14
15
16
17
18
19
20
21
173.4 qC
200.8 qC
79.7 qC
43.2 CH
29.4 CH2
173.4 qC
200.7 qC
79.7 qC
43.2 qC
29.5 CH2
2.41 m
2.41 m
1.79 ddd (14.6, 14.5, 4.0)
1.67 ddd (14.8, 3.6, 3.6)
1.38 ddd (14.3, 3.2, 3.2)
2.29 m
1.78 ddd (14.5, 14.4, 4.1)
1.67 ddd (14.7, 3.5, 3.5)
1.38 ddd (14.4, 3.4, 3.4)
2.32 ma
28.4 CH2
28.5 CH2
41.5 qC
156.4 qC
121.6 CH
163.4 qC
81.4 CH
18.6 CH3
12.2 CH3
119.3 qC
141.5 CH
109.7 CH
143.3 CH
28.1 CH3
20.6 CH3
66.3 CH
52.0 CH3
41.5 qC
156.6 qC
121.3 CH
163.3 qC
81.4 CH
18.6 CH3
12.2 CH3
119.3 qC
141.5 CH
109.7 CH
143.3 CH
28.1 CH3
20.5 CH3
66.1 CH
52.0 CH3
6.33 s
6.30 s
5.52 s
1.07 s
1.15 d (7.0)
5.52 s
1.07 s
1.14 d (7.0)
7.61 br s
6.50 br s
7.44 br s
1.17 s
1.10 s
6.27 s
7.61 br s
6.50 br s
7.44 br s
1.15 s
1.09 s
6.21 s
22
23
28
29
30
7-OMe
8-OH
30-Acyl
1′
3.70 s
3.77 s
3.70 s
3.79 s
172.3 qC
27.5 CH2
9.1 CH3
174.8 qC
33.8 CH
18.8 CH3
18.8 CH3
2′
2.25 m
1.09 t (7.5)
2.46 m
1.11 d (7.0)
1.09 d (7.0)
3′
4′
a Overlapped signals assigned by HSQC and HMBC spectra without designating multiplicity.
mg), piscidinol G (4.3 mg), protoxylocarpins A (9.1 mg) and D (41.7
mg), sapelin E acetate (3.2 mg), and xylogranatins N (41.5 mg) and O
(29.5 mg).
Absolute Configuration at C-2 in the 2-Methylbutyryl Group of
Hainangranatumins A and B (1 and 2). A portion of hainangrana-
tumin A (1, 5 mg) was dissolved in EtOH (0.5 mL) and treated with
5% KOH in H2O (1.0 mL), with stirring at room temperature for 24 h.
The reaction mixture was concentrated and partitioned between EtOAc
and H2O (3:1 v/v). After extraction with EtOAc (×3), the aqueous layer
was acidified with HCl to pH 3.0 and extracted again with CH2Cl2
(×3). The organic layer was combined and subjected to Sephadex LH-
20 CC (CH2Cl2-MeOH, 1:1) to provide 2-methylbutyric acid (0.7 mg),
which was dried over anhydrous Na2SO4 and identified on the basis of
its 1H NMR spectrum. The absolute configuration at C-2 was suggested

Limonoids from Xylocarpus granatum
Journal of Natural Products, 2010, Vol. 73, No. 10 1679
as R by its negative RD value {[R]25 -6.7 (c 0.03, Me2CO)}. In the
Foundation of China (20772135), the Research Foundation for Young
Talents from the South China Sea Institute of Oceanology, Chinese
Academy of Sciences (M.-Y.L., SQ200802), and the Beijing Natural
Science Foundation of China (7082060) is gratefully acknowledged.
D
same way, the absolute configuration of C-2 in the 2-methylbutyryl
group of hainangranatumin B was established as S by its positive RD
value {[R]25 +10.0 (c 0.04, Me2CO)}.
D
X-ray Crystallographic Analysis of Hainangranatumins A and
B (1 and 2). Colorless crystals of 1 and 2 were obtained in acetone.
Crystal data were obtained on a Bruker Smart 1000 CCD single-crystal
diffractometer with graphite-monochromated Mo KR radiation (λ )
0.71073 Å) and operating in the ω scan mode. The structure was solved
by direct methods (SHELXS-97) and refined using full-matrix least-
squares difference Fourier techniques. All non-hydrogen atoms were
refined anisotropically, and all hydrogen atoms were placed in idealized
positions and refined as riding atoms with the relative isotropic
parameters. Crystallographic data for 1 and 2 were deposited with the
Cambridge Crystallographic Data Centre with the deposition number
779982 and 779981, respectively. Copies of the data can be obtained,
free of charge, on application to the Director, CCDC, 12 Union Road,
Cambridge CB2 1EZ, UK [fax: +44(0)-1233-336033 or e-mail:
deposit@ccdc.cam.ac.uk].
Supporting Information Available: Copies of ESIMS of 1 and 2,
HR-TOFMS of 3-10, 1H NMR, 13C NMR, HSQC, HMBC, and
1
NOESY spectra of compounds 1-10, H-1H COSY spectra of 1-3,
5, and 8-10, and X-ray crystal data for 1 and 2. This material is
available free of charge via the Internet at http://pubs.acs.org.
References and Notes
(1) Wu, J.; Xiao, Q.; Xu, J.; Li, M. Y.; Pan, J. Y.; Yang, M. H. Nat.
Prod. Rep. 2008, 25, 955–981.
(2) Hu, W. M.; Wu, J. Open Nat. Prod. J. 2010, 3, 1–5.
(3) Li, M. Y.; Yang, S. X.; Pan, J. Y.; Xiao, Q.; Satyanandamurty, T.;
Wu, J. J. Nat. Prod. 2009, 72, 1657–1662.
(4) Li, M. Y.; Yang, X. B.; Pan, J. Y.; Feng, G.; Xiao, Q.; Sinkkonen, J.;
Satyanandamurty, T.; Wu, J. J. Nat. Prod. 2009, 72, 2110–2114.
(5) Wu, J.; Yang, S. X.; Li, M. Y.; Feng, G.; Pan, J. Y.; Xiao, Q.;
Sinkkonen, J.; Satyanandamurty, T. J. Nat. Prod. 2010, 73, 644–649.
(6) Cui, J.; Deng, Z.; Xu, M.; Proksch, P.; Li, Q.; Lin, W. HelV. Chim.
Acta 2009, 92, 139–150.
(7) Pudhom, K.; Sommit, D.; Nuclear, P.; Ngamrojanavanich, N.; Petsom,
A. J. Nat. Prod. 2009, 72, 2188–2191.
(8) Shen, L. R.; Dong, M.; Guo, D.; Yin, B. W.; Zhang, M. L.; Sha, Q. W.;
Huo, C. H.; Kiyota, H.; Suzuki, N.; Cong, B. Z. Naturforsch., C:
J. Biosci. 2009, 64, 37–42.
(9) Yin, B.; Huo, C.; Shen, L.; Wang, C.; Zhao, L.; Wang, Y.; Shi, Q.
Biochem. Syst. Ecol. 2009, 37, 218–220.
(10) Huo, C. H.; Guo, D.; Shen, L. R.; Yin, B. W.; Sauriol, F.; Li, L. G.; Zhang,
M. L.; Shi, Q. W.; Kiyota, H. Tetrahedron Lett. 2010, 51, 754–757.
(11) Zaridah, M. Z.; Idid, S. Z.; Omar, A. W.; Khozirah, S. J. Ethnophar-
macol. 2001, 78, 79–84.
(12) Murata, T.; Miyase, T.; Muregi, F. W.; Naoshima-Ishibashi, Y.;
Umehara, K.; Warashina, T.; Kanou, S.; Mkoji, G. M.; Terada, M.;
Ishih, A. J. Nat. Prod. 2008, 71, 167–174.
(13) Adesida, G. A.; Adesogan, E. K.; Taylor, D. A. H. Chem. Commun.
(London) 1967, 790–791.
(14) Rao, M. M.; Krishna, E. M.; Gupta, P. S.; Singh, P. P. Indian J. Chem.,
Sect. B: Org. Chem. Incl. Med. Chem. 1978, 16, 823–825.
(15) Saewan, N.; Sutherland, J. D.; Chantrapromma, K. Phytochemistry
2006, 67, 2288–2293.
(16) Kadota, S.; Marpaung, L.; Kikuchi, T.; Ekimoto, H. Chem. Pharm.
Bull. 1990, 38, 639–651.
(17) David, A. H.; Taylor, F. W. W. J. Chem. Soc., Perkin Trans. 1 1973, 1599–
1602.
(18) Lee, S. M.; Olsen, J. I.; Schweizer, M. P.; Klocke, J. A. Phytochemistry
1988, 27, 2773–2775.
(19) Yin, S.; Wang, X. N.; Fan, C. Q.; Lin, L. P.; Ding, J.; Yue, J. M. J.
Nat. Prod. 2007, 70, 682–685.
(20) Abdelgaleil, S. A. M.; Okamura, H.; Iwagawa, T.; Sato, A.; Miyahara,
I.; Doe, M.; Nakatani, M. Tetrahedron 2001, 57, 119–126.
(21) Ng, A. S.; Fallis, A. G. Can. J. Chem. 1979, 57, 3088–3089.
(22) Balakrishna, K.; Sukumar, E.; Connolly, J. D. Nat. Prod. Sci. 2003, 9,
192–194.
Crystal Data of 1: monoclinic, C32H40O10, space group P21 with a )
8.1004(4) Å, b ) 16.0763(8) Å, c ) 11.7961(6) Å, V ) 1514.45(13) Å3,
Z ) 2, Dcalcd ) 1.282 g/cm3, m ) 0.095 mm-1, and F(000) ) 624. Crystal
size: 0.46 × 0.42 × 0.41 mm3. Independent reflections: 3034 [Rint
)
0.0164]. The final indices were R1 ) 0.0336, wR2 ) 0.0892 [I > 2σ(I)].
Crystal Data of 2: monoclinic, C32H40O10, space group P21 with a )
7.9418(4) Å, b ) 16.2706(9) Å, c ) 11.6249(6) Å, V ) 1491.53(14) Å3,
Z ) 2, Dcalcd ) 1.302 g/cm3, m ) 0.096 mm-1, and F(000) ) 624. Crystal
size: 0.46 × 0.42 × 0.32 mm3. Independent reflections: 3355 [Rint
)
0.0197]. The final indices were R1 ) 0.0352, wR2 ) 0.0993 [I > 2σ(I)].
Hainangranatumin A (1): colorless crystals (acetone); mp 125-127
°C; [R]25 -28.6 (c 2.47, Me2CO); UV (MeOH) λmax 210.9 nm; for H
1
D
and 13C NMR data, see Tables 1 and 2; LR-ESIMS m/z 607.1 [M + Na]+.
Hainangranatumin B (2): colorless crystals (acetone); mp 130-131
°C; [R]25 -22.8 (c 0.88, Me2CO); UV (MeOH) λmax 210.9 nm; for H
1
D
and 13C NMR data, see Tables 1 and 2; LR-ESIMS m/z 607.1 [M + Na]+.
Hainangranatumin C (3): white, amorphous powder; [R]25D -11.3
1
(c 5.99, Me2CO); UV (MeOH) λmax 212.5 nm; for H and 13C NMR
data, see Tables 1 and 2; HR-TOFMS m/z 579.2185 [calcd for
C30H36O10Na [M + Na]+, 579.2206].
Hainangranatumin D (4): white, amorphous powder; [R]25D -37.1
1
(c 0.62, Me2CO); UV (MeOH) λmax 212.1, 236.9 nm; for H and 13C
NMR data, see Tables 1 and 2; HR-TOFMS m/z 567.2173 [calcd for
C29H36O10Na [M + Na]+, 567.2206], HR-TOFMS m/z 545.2371 [calcd
for C29H37O10 [M + H]+, 545.2387].
Hainangranatumin E (5): white, amorphous powder; [R]25D +8.8
1
(c 0.08, Me2CO); UV (MeCN) λmax 205.1 nm; for H and 13C NMR
data, see Tables 1 and 2; HR-TOFMS m/z 639.2435 [calcd for
C32H40O12Na [M + Na]+, 639.2417].
Hainangranatumin F (6): white, amorphous powder; [R]25D -55.6
1
(c 0.48, Me2CO); UV (MeCN) λmax 210.9 nm; for H and 13C NMR
data, see Table 3; HR-TOFMS m/z 581.2319 [calcd for C30H38O10Na
[M + Na]+, 581.2363], HR-TOFMS m/z 559.2549 [calcd for C30H39O10
[M + H]+, 559.2543].
(23) Naidoo, D.; Mulholland, D. A.; Randrianarivelojosia, M.; Coombes,
P. H. Biochem. Syst. Ecol. 2003, 31, 1047–1050.
(24) Gan, L.-S.; Wang, X.-N.; Wu, Y.; Yue, J.-M. J. Nat. Prod. 2007, 70,
1344–1347.
(25) Okorie, D. A. H. T. J. Chem. Soc. C 1970, 211–213.
(26) Cui, J. X.; Wu, J.; Deng, Z. W.; Proksch, P.; Lin, W. H. J. Nat. Prod.
2007, 70, 772–778.
(27) Wu, J.; Xiao, Q.; Huang, J. S.; Xiao, Z. H.; Qi, S. H.; Li, Q. X.;
Zhang, S. Org. Lett. 2004, 6, 1841–1844.
(28) Wu, J.; Xiao, Q.; Zhang, S.; Li, X.; Xiao, Z.; Ding, H.; Li, Q.
Tetrahedron 2005, 61, 8382–8389.
(29) Yin, S.; Fan, C. Q.; Wang, X. N.; Lin, L. P.; Ding, J.; Yue, J. M.
Org. Lett. 2006, 8, 4935–4938.
(30) Wu, J.; Zhang, S.; Bruhn, o.; Xiao, Q.; Ding, H. Chem.sEur. J. 2008,
14, 1129–1144.
(31) Meyers, A. I.; Knaus, G.; Kamata, K. J. Am. Chem. Soc. 1974, 96, 268–
270.
Hainangranatumin G (7): white, amorphous powder; [R]25D +135.3
1
(c 0.17, Me2CO); UV (MeCN) λmax 212.5, 273.7, 315.1 nm; for H
and 13C NMR data, see Table 3; HR-TOFMS m/z 478.2239 [calcd for
C28H31O6NNa [M + H]+, 478.2230].
Hainangranatumin H (8): white, amorphous powder; [R]25D -56.4
1
(c 0.80, Me2CO); UV (MeOH) λmax 209.8 nm; for H and 13C NMR
data, see Table 4; HR-TOFMS m/z 783.2464 [calcd for C37H44O17Na
[M + Na]+, 783.2476], HR-TOFMS m/z 761.2676 [calcd for C37H45O17
[M + H]+, 761.2657].
Hainangranatumin I (9): white, amorphous powder; [R]25D +83.5
1
(c 0.31, Me2CO); UV (MeOH) λmax 240.7 nm; for H and 13C NMR
data, see Table 5; HR-TOFMS m/z 579.2239 [calcd for C30H36O10Na
[M + Na]+, 579.2206].
Hainangranatumin J (10): white, amorphous powder; [R]25D +81.4
1
(c 1.08, Me2CO); UV (MeOH) λmax 240.7 nm; for H and 13C NMR
(32) Brechbuhler, S.; Buc¨hi, G.; Milne, G. J. Org. Chem. 1967, 32, 2641–2642.
(33) Korver, O.; Gorkom, M. V. Tetrahedron 1974, 30, 4041–4048.
(34) Wu, J.; Zhang, S.; Xiao, Q.; Li, Q.; Huang, J.; Long, L.; Huang, L.
Tetrahedron Lett. 2004, 45, 591–593.
(35) Wu, J.; Xiao, Q.; Zhang, S.; Li, X.; Xiao, Z. H.; Ding, H. X.; Li,
Q. X. Tetrahedron 2005, 61, 8382–8389.
data, see Table 5; HR-TOFMS m/z 593.2352 [calcd for C31H38O10Na
[M + Na]+, 593.2363].
Acknowledgment. Financial support for this work from the Impor-
tant Project of Chinese Academy of Sciences (KSCX2-YW-R-093),
the National High Technology Research and Development Program of
China (863 Program) (2007AA09Z407), the National Natural Science
NP100395W