全 文 ：Journal of Chinese Pharmaceutical Sciences 2000, 9 (4)170
Five Polyoxygenated Cyclohexenes from Uvaria grandiflora**
Liao Yonghong1, Zou Zhongmei1*, Guo Jian1, Xu Lizhen1, Zhu Min2 and Yang Shilin1
1. Institute of Medicinal Plant Development, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100094;
2. Department of Pharmacy, The University of Chinese Hong Kong, Shatin, New Territories, Hong Kong
Received August 7, 2000; Accepted September 27, 2000
Abstract Five new polyoxygenated cyclohexenes, uvarigranones A (2), B (3), C (6) D (7) and
uvarigranol J (5) were isolated from the stems of Uvaria grandiflora. Their structures were established on
the basis of spectral data and comparison with known compounds. Among them, the absolute
configurations of 2 , 5 , 6 and 7 were determined from their CD spectra.
Key words Uvaria grandiflora; Polyoxygenated cyclohexene; Uvarigranone A; B; C; D and
uvarigranol J.
Introduction
Previous phytochemical investigations of the stems
and leaves and the roots of Uvaria grandiflora(1~4) resulted
in the isolation of polyoxygenated cyclohexenes.
Zeylenone (1), a polyoxygenated cyclohexene from the
stems and leaves, showed remarkable inhibition of thy-
midine and uridine transport in Ehrlich carcinoma cells
and cytotoxicity to cultured cancer cells with equal po-
tency against sensitive and multidrug resistant cell
lines (1). To further investigate the compounds of this
type, we have isolated five new polyoxygenated
cyclohexenes, uvarigranol J(5), uvarigranones A(2), B(3),
C(6) and D(7) from the stems of U. grandiflora. This paper
describes the characterization of these new compounds.
1. R1=R3=Bz, R2=H 4 . R1=Ac, R2=H 6 . R=Et
2. R1=R2=Bz, R3=H 5 . R1=H, R2=Ac 7 . R=H
3. 3 . R1=H, R2=R3=Bz
HMBC: NOESY:
Fig. 1. Key HMBC and NOESY Correlations of 6.
Results and Discussion
Compound 2 showed protonated molecular ion peak
at m/z 383 [M+H]+ in its positive-ion FABMS and its
molecular formula was established as C21H18O7. The IR
spectrum of 2 suggested the presence of a hydroxyl
group (3420 cm-1), a ketone group (1720 cm-1), ester
groups (1700 and 1690 cm-1) and a monosubstituted
phenyl ring (1600, 1580, 1450 and 710 cm-1). The 1H and
13C NMR spectra (Table 1 and 2) of 2 were very similar to
those of 1(1). The presence of two benzoyl groups was
confirmed by the aromatic proton signals between d
7.37~7.95 (10H, m). When the 1H NMR spectrum of 2 was
compared with that of 1, the downfield shift of H-2 (d
5.66, dd, J=4.0, 1.1 Hz) and the upfield shift of H-3 (d 4.80,
td, J=4.0,1.1 Hz) of 2 indicated that the two benzoyl
groups were located at C-2 and C-7.
The coupling constant 4.0 Hz of J3,4 showed that H-
3 was equatorial (allylic couplings had equatorial values
of about 4 Hz or axial values of 1.9~2.6 Hz)(5, 6). In addi-
tion, there were very similar correlations in the NOESY
spectra of 1 and 2. Compound 2 had a strong correlation
between d 4.84 (H-7) and d 5.66 (H-2) and weak corre-
lation between d 4.72 (H-7) and d 5.66 (H-2) which
showed a cis- vicinal relationship between C1-OH and C2-
Obz(3). On the basis of the above data, compounds 1 and
2 had the same relative stereochemistry. The absolute
configuration shown in 2 was based on its CD spectrum
which exhibited a strong positive Cotton effect (l241 nm,
Äe+120.61) and a negative Cotton effect (l221 nm,
Äe-43.42) of two benzoyl groups.
Compound 5 was obtained as a white powder. The
FABMS spectrum gave a [M+H]+ at m/z 427, indicating a
molecular formula of C23H22O8. Its IR spectrum exhibited
the presence of a hydroxyl group (3440 cm-1), ester
groups (1740, 1710 and 1700 cm-1) and a monosubstituted
phenyl ring (1600, 1580, 1450 and 710 cm-1), which were
similar to those of uvarigranol B (4)(3). The NMR spectra
R2O
OH
R1O
R3O
O R1O
OH
BzO
BzO
OR2
HO
OH
BzO
BzO
O
OR
12
3
4
5
6
7
HO
Hb
PhCOO
Ha
O
H
OH
H
H
H
H
OCH2CH3
OCOPh
1
2
4
5
6
7
*Author to whom correspondence should be addressed.
**This project was supported by the National Natural Sciences Foundation of China and the Chinese Doctoral Grants from the Ministry
of Science and Technology, China.
Journal of Chinese Pharmaceutical Sciences 2000, 9 (4) 171
(Tables 1 and 2) of 5 were very similar to those of 4. The
difference was the upfield shift of H-2 (d 4.24, d, J=6.1Hz)
and the downfield of H-6 (d 5.68, dt, J=4.0, 0.6 Hz) in 5
which indicated that the acetyl group of 5 was located at
C-6 instead of at C-2 in 4. On the basis of the comparison
of the coupling constants of 4 and 5 (Table 1), as well the
similar strong correlation of NOESY spectrum between H-
7 and H-2, 5 was established to have the same relative
stereochemistry as 4(3).
The absolute configuration shown in 5 was based on
comparison with 4. The two benzoyl groups at C-3 and C-
7 had the same relative stereochemistry and their CD
spectra exhibited a similar strong negative Cotton effect
(4, l227 nm, Äe-160.38; 5, l235 nm, Äe-71.97). Thus, the
absolute configuration of 5 was identical to that of 4.
Compound 6 showed an molecular ion peak at m/z
429 [M+H]+ in its positive-ion FABMS. Thus its mo-
lecular formula was determined to be C23H24O8. Its IR
spectrum exhibited the presence of a hydroxyl group
(3430 cm-1), ester groups (1730, 1710 and 1700 cm-1) and a
monosubstituted phenyl ring (1600, 1580, 1450 and 710
cm-1). The aromatic proton signals between d 7.37~7.95
(10H, m) in the 1H NMR (Table 1) spectrum of 6 indicated
the presence of two benzoyl groups. The 13C-NMR
(Table 2) spectrum showed 21 carbon signals including
two benzoyl groups and an ethoxyl group. These
observations suggested that 6 was a polyoxygenated
cyclohexane. The 1H-1H COSY spectrum of 6 showed
correlation for H2-5 (d 3.18, dd, J=17.6, 10.0 Hz and d 2.96,
ddd, J=17.6, 4.2, 1.0 Hz) and H-4 (d 4.08, dt, J=10.0, 4.2 Hz),
which was in turn connected to H-3 (d 5.92, ddd, J=5.0,
4.2, 1.0 Hz). The latter was proton coupled to H-2 (d 4.29,
d, J=5.0 Hz). The HMBC spectrum (Fig. 1) of 6 showed
correlations between d 5.92 (H-3) and d 165.7 (C=O) and
between d 5.13 and 4.39 (H2-7) and d 166.2 (C=O), which
suggested that the two benzoyl groups were at C-3 and
C-7, respectively, while the correlation between d 4.08 (H-
4) and d 65.3 (OCH2Me) indicated that the ethoxyl group
was located at C-4.
The relative stereochemistry of 6 was deduced from
a combination of coupling constant-analyses and the
NOESY spectrum (Fig. 1). A coupling constant 10.0 Hz of
J4,5 suggested that H-4 was axial and the ethoxyl group at
C-4 was equatorial; 4.2 Hz of J3,4 indicated that H-3 was
equatorial and the benzoyl group at C-3 was axial. The
axial orientation of C-7 was confirmed by observing the
NOE correlation between Ha-7 and Hb-5. The strong NOE
correlation between Hb-7 and H-2 and the weak
correlation between Ha-7 and H-2 demonstrated that the
cis-vicinal relationship between C1-OH and C2-OH. By
means of the CD spectrum, a negative Cotton effect (l290
nm, Äe-4.32) of the ketone group and a positive Cotton
effect (l233 nm, Äe+61.21) as well a negative Cotton
effect (l220 nm, Äe-18) of the two benzoyl groups
confirmed the absolute configuration shown in 6.
Compound 7 gave a protonated molecular ion peak
at m/z 401 [M++H] in its positive ion FAB mass spec-
trometry corresponding to C21H20O8. Compound 7 gave
rise to a 1H NMR spectrum (Table 1) and a 13C NMR
spectrum (Table 3) similar to that of 6, which suggested a
structure closely related to 6. The only difference was
that compound 7 had no ethoxyl group at C-4. There
were very similar coupling constants (Table 1) and
NOESY spectra which showed identical correlations
between Ha-7 and Hb-5, Hb-7 and H-2 as well as Ha-7 and
H-2. This showed that the relative stereochemistry of 7
was identical to that of 6. The CD spectrum of 7, which
exhibited a negative Cotton effect (l290 nm, Äe-3.95) of
the ketone group and a positive Cotton effect (l233 nm,
Äe+71.15) as well a negative Cotton effect (l220 nm, Äe-
14.23) of the two benzoyl groups, confirmed that the
absolute configuration of 7 was the same as that of 6.
Compound 3 was obtained as a mixture with 1 (1:1).
The NMR (1H, 13C, COSY, HMQC and HMBC) spectra
indicated that 3 had a similar structure to 1. Their
difference was the position of a benzoyl group. The
HMBC spectrum of 3 showed correlations between d 6.11
(H-3) and d 165.5 (C=O) and between d 5.84 (H-2) and d
165.3 (C=O), which suggested that the two benzoyl
groups were at C-2 and C-3. The comparison of the
coupling constants of 1 and 3 (Table 1) and the strong
correlation between d 4.00 (H-7) and d 5.84 (H-2) in the 1H
-1H COSY spectrum, indicated that 3 had the same
relative stereochemistry as 1(1).
Experimental
Optical rotations were determined on a Perkin-Elmer
241 polarimeter. The NMR spectra (1H at 400 MHz and 13C
at 100 MHz) were recorded on a Bruker ARX 400
spectrometer in CDCl3 using TMS as internal standard.
UV spectra were obtained on a Philips PYE Union PU8800
spectrophotometer, and IR spectra on a Perkin-Elmer
983G instrument. EIMS (70eV) and FABMS were
determined on a VG ZAB-2F mass spectrometer. Mps
were uncorrected and were taken on a Fisher-John hot-
stage apparatus.
Plant
The stems of U. grandiflora were collected in Hainan
Province of China in July 1992, and identified by
Professors Lin Shouqun and Lian Wenyan of this
institute.
Journal of Chinese Pharmaceutical Sciences 2000, 9 (4)172
Table 1. 1H NMR data for Compounds 2~7 (d , JHz, in CDCl3)
Position 2 3 4 5 6 7
2 5.66,dd,
(4.0,1.1)
5.84,d,(5.1) 5.72,d,(6.8) 4.24,d,(6.1) 4.29,d,(5.0) 4.30,d,(5.0)
3 4.80,td,
(4.0,1.1)
6.11,ddd,
(5.1,3.4,1.4)
5.81,dddd,
(6.8,2.5,1.7,0.6)
5.77,dddd,(6.1,
2.5,1.6,0.6)
5.92,ddd,
(5.0,4.2,1.0)
5.82,ddd,
(5.0,3.8,1.0)
4 6.93,ddd,
(10.1,4.0,1.1)
7.00,dd,
(10.2,3.4)
5.92,ddd,
(10.0,2.5,0.6)
5.98,dd
(10.0,2.5)
4.08,dt,
(10.0,4.2)
4.55,ddd,
(10.0,4.6,3.8)
5 6.27,dd,
(10.1,1.1)
6.32,dd,
(10.2,1.4)
6.00,ddd,
(10.0,4.0,1.7)
6.05,ddd
(10.0,4.0,1.6)
Ha: 3.18,dd,
(17.6,10.0)
Hb: 2.96,ddd,
(17.6,4.2,1.0)
3.11,dd,
(17.6,10.0)
3.02,ddd,
(17.6,4.6,1.0)
6 4.34,dt,
(4.0,0.6)
5.68,dt, (4.0,0.6)
7-Ha
-Hb
4.84,d,(11.8)
4.72,d,(11.8)
4.00,s
4.00,s
4.76,d,(12)
4.56,d,(12)
4.77
4.62
5.13,d,(11.8)
4.39,d,(11.8)
5.09,d,(11.8)
4.42,d,(11.8)
OAc 2.07,s 2.04,s
OBz 7.37~7.97,
m,10H
7.39~8.03,
m,10H
7.35~8.02,
m,10H
7.40~8.05, m,
10H
7.48~8.06, m,10H
OEt 3.54~3.64,m
1.1,t,(6.8)
Extraction and isolation
The stems (9 kg) of U. grandiflora , which were cut
into small pieces, were extracted with 95% EtOH for three
times under reflux and the combined extract was evapo-
rated under reduced pressure at 55 oC to give 380 g resi-
due. The residue was chromatographed over a Si gel
column, which was eluted with a petroleum ether-Me2CO-
MeOH gradient to afford 10 combined fractions (F1-F10),
among which F3-F7 contained polyoxygenated
cyclohexenes (petroleum ether: Me2CO, Gradient from 7:1
to 3:2). Purification of F3 by Si gel column [petroleum
ether-EtOAc (2:1)] yielded grandifloracin (200 mg)(1),
Uvarigranol B (4, 800 mg) and Uvarigranol J (5, 150 mg).
F4 was chromatographed on Si gel using a gradient of
30% EtOAc in petroleum ether to afford Uvarigranone A
(2, 120 mg) and the mixture of 1 and 3. Recrystallization
of F5 gave zeylenone (1, 8 g). Uvarigranone C (6, 80 mg)
and Uvarigranone D (7, 50 mg) were obtained from
purification of F6. Purification of F7 by Sephadex LH-20
yielded zeylenol (2 g)(1).
Uvarigranone A (2). White solid, mp 72 oC, [a] 20D +
70.4° (CHCl3; c0.47); UV lmax (CHCl3) nm: 202, 211, 240,
271; CD lmax(CH3CN) nm: 241 (Äe+120.61), 221 (Äe-43.42),
IRnmax (KBr) cm-1: 3420 (OH), 1720, 1700, 1690, 1600,1580,
1450, 1270, 1120, 710. EIMS m/z (rel int): 201 (5), 122 (10),
105 (100), 97 (3), 77 (28); FABMS m/z (rel int): 383[M+H]+
(12), 365 (10), 261 (8), 105 (100), 77 (8); 1H NMR see Table
1; 13C NMR see Table 2.
Uvarigranone B (3). 1H NMR: see Table 1; 13C NMR:
see Table 2.
Uvarigranol J (5). White powder, mp 100 oC, [a] 20D +
61.2° (CHCl3; c0.041); UV lmax (CHCl3) nm: 207, 212, 240,
271; CD lmax (CH3CN) nm: 235 (Äe-71.97); IRnmax (KBr)
cm-1: 3440 (OH), 1740, 1710, 1700, 1600,1580, 1450, 1280,
1120, 710. EIMS m/z (rel int): 409 (1), 305(3), 203 (10), 163
(2), 122 (4), 105 (100), 77 (18); FABMS m/z (rel int): 427
[M+H]+ (32), 409 (5), 305 (95), 191 (8), 105 (100), 77 (8); 1H
NMR see Table 1; 13C NMR see Table 2.
Table 2. 13C NMR Chemical Shifts (d ) for Compounds
2~7(CDCl3)
Position 2 3 4 5 6 7
1 76.2 77.2 75.2 74.7 79.4 79.1
2 75.4 72.4 71.6 71.0 71.3 71.2
3 67.2 68.7 71.3 73.4 70.2 67.6
4 146.6 143.8 127.3 126.6 74.0 73.0
5 127.1 128.5 129.2 128.7 40.6 42.1
6 195.9 197.0 69.3 71.6 205.4 205.0
7 66.0 64.2 66.3 66.4 65.4 65.2
Acetyl C=O 170.5 171.7
CH3 20.8 20.5
a Benzoyl,C=O 166.0 165.3 166.0 165.8 165.7 166.4
1 128.9 129.3 129.1 129.4 129.2 129.1
2/6 129.8 129.9 129.7 129.7 129.8 129.8
3/5 128.4 128.6 128.4 128.5 128.5 128.5
4 133.3 133.5 133.3 133.5 133.5 133.5
aBenzoyl,C=O 166.1 165.5 167.3 167.0 166.2 166.4
1 129.3 128.7 129.4 129.8 129.4 129.1
2/6 129.9 129.8 129.8 129.9 129.9 129.9
3/5 128.6 128.5 128.5 128.6 128.7 128.8
4 133.7 133.7 133.6 133.6 133.6 133.9
4-OEt CH 2 65.3
CH3 15.3
aAssignments may be interchanged.
Uvarigranone C(6). White solid, mp 146 oC, [a] 20D -
5.9° (CHCl3; c0.42); UV lmax (CHCl3) nm: 205, 212, 240,
271; CD lmax (CH3CN) nm: 290 (Äe-4.32), 233 (Äe+61.21),
220 (Äe-18); IR nmax (KBr) cm-1: 3420 (OH), 2960, 1730,
1710, 1700, 1600,1580, 1450, 1280, 1120, 710. EIMS m/z (rel
Journal of Chinese Pharmaceutical Sciences 2000, 9 (4) 173
int): 367 (4), 191(30), 163 (8), 122 (12), 105 (100), 77 (38);
FABMS m/z (rel int): 429[M+H]+ (12), 411 (8), 367 (2), 307
(4), 105 (100), 77 (10); 1H NMR see Table 1; 13C NMR see
Table 2.
Uvarigranone D (7). White powder, mp 54 oC,
[a] 20D +11.4° (CHCl3; c0.43); UV lmax (CHCl3) nm: 202, 212,
240, 272; CD lmax (CH3CN) nm: 290 (Äe-3.93), 233
(Äe+71.15), 220 (Äe-14.23); IR nmax (KBr) cm-1: 3440 (OH),
1730, 1710, 1700, 1600,1580, 1450, 1280, 1120, 710. EIMS
m/z (rel int): 261 (1), 163 (8), 122 (15), 105 (100), 77 (38);
FABMS m/z (rel int): 401[M+H]+ (45), 383 (10), 279 (11),
261(8), 105 (100), 77 (8); 1H NMR see Table 1; 13C NMR
see Table 2.
Acknowledgements This research project was supported by the National Natural Sciences Foundation of China and
the Chinese Doctoral Grants from the Ministry of Science and Technology, China. The authors would like to thank Mr.
Zhu Pin for collecting the plant materials, Prof. Lu Mujian for recording NMR spectra, Prof. Cong Puzhu for EIMS, Mr.
Wang Jie for FABMS and Ms. Liu Huiling for UV and IR spectra.
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2 Liao YH, Xu LZ Yang SL and Sun NJ, Chin Tradit Herb Drugs, 1996, 27: 524
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大花紫玉盘中的多氧取代环己烯衍生物
廖永红1 邹忠梅1 郭剑1 徐丽珍1 朱敏2 杨世林1
1. 中国医学科学院中国协和医科大学药用植物研究所 北京 100094
2. 香港中文大学药学院 香港
摘要 从大花紫玉盘茎中分离得到5个新的多氧取代环己烯衍生物 根据光谱分析及与已知化合物比较确定了它们的结
构 并分别命名为大花紫玉盘酮A 2 B 3 C 5 D 7 及大花紫玉盘醇J 5 同时 应用CD谱测定了化
合物2 5 6 7 的绝对构型
关键词 大花紫玉盘 多氧取代环己烯 大花紫玉盘酮A B C D 大花紫玉盘醇 J