Pregnane and pregnane glycosides from the malagasy Plant, Cynanchum aphyllum

Article abstract of DOI:10.1248/cpb.50.1031

A new 8,14-seco-pregnane type of steroid called cynaphyllogenin and its eight glycosides, cynaphyllosides A-H, were isolated from the aerial parts of Cynanchum aphyllum. The structures of these compounds were elucidated based on chemical and spectroscopic

Full text of DOI:10.1248/cpb.50.1031

August 2002
Chem. Pharm. Bull. 50(8) 1031—1034 (2002)
1031
Pregnane and Pregnane Glycosides from the Malagasy Plant,
Cynanchum aphyllum
Tripetch KANCHANAPOOM,a,b Ryoji KASAI,a Kazuhiro OHTANI,a Marta ANDRIANTSIFERANA,c and
,a
*
Kazuo YAMASAKI
a Division of Medicinal Chemistry, Graduate School of Biochemical Sciences, Hiroshima University; 1–2–3 Kasumi,
Minami-ku, Hiroshima 734–8551, Japan: b Department of Pharmaceutical Botany and Pharmacognosy, Faculty of
c
Pharmaceutical Sciences, Khon Kaen University; Khon Kaen 40002, Thailand: and Laboratoire de Chimie Organigue,
Produits Naturels, Universite’ d’Antananarivo; Madagascar. Received February 12, 2002; accepted April 15, 2002
A new 8,14-seco-pregnane type of steroid called cynaphyllogenin and its eight glycosides, cynaphyllosides
A—H, were isolated from the aerial parts of Cynanchum aphyllum. The structures of these compounds were elu-
cidated based on chemical and spectroscopic evidence.
Key words cynaphyllosides A—H; 8,14-seco-pregnane; Cynanchum aphyllum; cynaphyllogenin; pregnane glycosides; Asclepi-
adaceae
Cynanchum aphyllum L. (Asclepiadaceae, Malagasy name: signal at d 31.1 as well as a singlet methyl proton signal at d
Vahimasy) is a leafless and succulent shrub which secretes 2.30 were characteristic of a methyl ketone group. These re-
yellow latex. No phytochemical investigation has been car- sults demonstrated that 1 is a 3b,5b-dihydroxy-8,14-seco-
ried out on this species. In our continuing study on the chem- pregn-6-ene-8,14,20-trione type of compound. The configu-
ical constituents of Malagasy plants, we isolated a pregnane ration of the side chain at C-17 was as follows. The proton
called cynaphyllogenin (1) and its eight glycosides, cyna- signal of H-17 (d 2.93) was observed as double doublet with
phyllosides A—H (2—9), from the aerial parts of this plant. the coupling constants 10.5 and 6.4 Hz.4) The nuclear Over-
The present paper deals with the isolation and structural de- hauser enhancement (NOE) spectrum of 1 provided further
termination of these compounds.
Table 1. 1H- and 13C-NMR Spectral Data of Cynaphyllogenin (1) in Pyri-
dine-d5
The methanolic extract of the aerial part of C. aphyllum
was partitioned with n-hexane, Et2O, and H2O. The Et2O and
H2O soluble fractions were purified by column chromatogra-
phy and then by preparative HPLC to afford compounds 3, 5
and 6 from the former fraction and 1, 2, 4, and 7—9 from the
latter fraction.
No.
1
1H
13C
HMBC (H→C)
1.57 m
25.7
28.6
5
10
2.21 td (Jϭ14.2, 3.2 Hz)
1.74 m
2
The molecular formula of cynaphyllogenin (1) was deter-
mined as C30H36O7 by negative high resolution (HR)-FAB
mass spectrometry and 13C-NMR spectral data. The 13C-
NMR spectrum of 1 revealed the presence of 30 signals:
three methyls, six methylenes, four methines (two of them
bearing an oxygen atom), three quarternary (one of them
bearing an oxygen atom), two di-substituted double bonds,
one mono-substituted benzene ring and four carbonyl (three
ketones and one ester). Of these signals, nine due to the ester
carbonyl, benzene ring and one of the di-substituted double
bond carbons were easily assigned to cinnamoyl moiety. The
double bond of this moiety was determined to be trans-con-
figuration from the coupling constants of its two olefinic pro-
tons.
1.87 m
4.25 m
1.95 dd (Jϭ14.4, 2.5 Hz)
2.09 br d (Jϭ14.4 Hz)
3
4
66.3
38.5
2, 3, 5, 10
5
6
7
8
9
74.9
154.5 4, 10
126.9 5, 9
201.9
47.9 5, 8, 10, 11, 12, 19
46.0
26.4 8, 12
6.70 d (Jϭ10.1 Hz)
5.93 d (Jϭ10.1 Hz)
3.29 br d (Jϭ10.6 Hz)
10
11
1.77 br dd (Jϭ14.3, 9.7 Hz)
2.47 br dd (Jϭ14.3, 10.6 Hz)
5.99 br d (Jϭ9.7 Hz)
8, 9
12
13
14
15
71.3 9, 11, 13, 17, 18, 1Ј
57.4
215.7
35.2 14, 16, 17
2.32 m
3.09 ddd (Jϭ20.2, 9.8, 7.6 Hz)
14,17
The other 21 carbon signals were correlated by hetero-
nuclear multiple-bond connectivity (HMBC) experiments
(Table 1). From this result and the chemotaxonomical fact
that the presence of pregnanes and their glycosides in this
genus are well known,1,2) we assumed that 1 is an 8,14-seco-
pregnane. This type of pregnane has already been isolated as
a glycoside called sarcovimiside B (10) from Sarcostemma
viminale.3) The carbon signals due to the pregnane skeleton
(A and B-rings, C-11 and C-12) of 1 were very similar to
those of the aglycone of 10, lacking the acetyl and carbinyl
carbon signals attributed to the side chain of 10 and instead
had a carbonyl signal for ketone. Moreover, chemical shifts
of this carbonyl carbon signal at d 207.5 and a methyl carbon
16
1.90 m
2.56 m
2.93 dd (Jϭ10.5, 6.4 Hz)
1.70 s
1.00 s
20.3 13,14
17,20
17
18
19
20
21
1Ј
2Ј
3Ј
4Ј
5Ј,9Ј
59.2 12, 13, 14, 15, 18, 20
17.1 12, 13, 14, 17
17.9 1, 5, 9, 10
207.5
31.1 17, 20
167.0
118.3
146.0
134.9
128.7
2.30 s
6.83 d (Jϭ16.0 Hz)
8.01 d (Jϭ16.0 Hz)
6Ј,8Ј
7Ј
129.3
130.8
To whom correspondence should be addressed. e-mail: kazuoy@hiroshima-u.ac.jp
© 2002 Pharmaceutical Society of Japan

1032
Vol. 50, No. 8
Fig. 1
confirmation of the strong correlation between H-18 (d 1.70) units of cymarose and one terminal thevetose in 5. Moreover,
and H-21 (d 2.30). The position of the cinnamoyl moiety on the order of these sugar sequences was confirmed by obser-
C-12 was determined from HMBC experiment (Table 1).
vation of the peak at m/z 795 [(MϪterminal Glc or The)Ϫ] in
These results led to the formulation of cynaphyllogenin (1) the negative FAB-MS spectra of both 4 and 5. Therefore, the
as 12(R)-O-cinnamoyloxy-3b,5b-dihydroxy-8,14-seco-17b- structures of 4 and 5 were formulated as shown in Fig. 1.
pregn-6-ene-8,14,20-trione.
Cynaphylloside E (6) had a molecular formula C50H70O17.
Cynaphyllosides A—H (2—9) were assigned as the glyco- The component sugars were identified as cymarose, digitox-
1
sides of cynaphyllogenin with different sugar moiety (Tables ose and thevetose by acid hydrolysis. The H- and 13C-NMR
1
2, 3) from the 13C- and H-NMR data. Comparison of the spectra indicated the presence of one unit of each of b-cy-
13C-NMR spectra of 2—9 showed glycosylation shifts at C-3, marose, b-digitoxose and a terminal b-thevetose. Mild acid
C-2 and C-4 indicating that the sugar moiety of all these hydrolysis of 6 yielded 2 as a partially hydrolyzed product
compounds is located at C-3 of the aglycone.
together with 1 indicating that the sugar sequence is –cy-
The component sugars isolated from acid hydrolysate of marose–thevetose–digitoxose. The linkage of the thevetose to
the glycosides were all confirmed as D-form by measurement C-4 of the digitoxose was determined from observation of
of their optical rotations.5) The linkages of all sugars were as- the downfield shift of C-4 and the upfield of C-3 and C-5 of
signed to be in the b-form base on the coupling constant of digitoxose in the 13C-NMR spectrum of 6. Therefore, the
anomeric protons in 1H-NMR spectra.
structure of 6 was assigned as shown in Fig. 1.
The molecular formula of cynaphylloside A (2) was deter-
Cynaphylloside F (7) had a molecular formula C57H82O22.
1
mined as C37H48O10. The 1H- and 13C-NMR spectra indicated The H- and 13C-NMR spectra of 7 showed four anomeric
the presence of one monosaccharide unit. Acid hydrolysis of signals. Acid hydrolysis gave cymarose, thevetose and glu-
2 gave cymarose. Accordingly, the structure of 2 was eluci- cose. The electron impact (EI)-MS of its acetyl derivative
dated as shown in Fig. 1.
displayed a fragment ion at m/z 331 due to terminal b-glu-
Cynaphylloside B (3) had a molecular formula C43H58O15. copyranose. This was confirmed by obtaining 5 on enzymatic
1
Acid hydrolysis provided cymarose and glucose. In the H- hydrolysis of 7 with crude hesperidinase. The position of the
NMR spectrum of 3, two anomeric protons were observed. terminal glucose at C-4 of the thevetose moiety was deter-
The 13C-NMR spectrum showed the presence of a terminal mined by observation of the glycosylation shifts for the sig-
b-D-glucopyranosyl unit. Consequently, the structure of 3 nals due to C-4 (ϩ7.2 ppm), C-3 (Ϫ2.0 ppm) and C-5
was determined as shown in Fig. 1.
(Ϫ0.9 ppm) in comparison with the 13C-NMR spectra of 5
Cynaphylloside C (4) and cynaphylloside D (5) had the and 7. Based on these results, the structure of 7 was formu-
molecular formula C50H70O18 and C51H72O17, respectively. lated as shown in Fig. 1.
Cymarose and glucose were detected from the acid hy-
The molecular formula of cynaphyllosides G (8) and H (9)
drolysate of 4, and cymarose and thevetose from that of 5. was determined as C56H80O22 and C62H90O27, respectively. By
1
The H- and 13C-NMR spectra revealed the presence of two the same method as used for 7, the structures of 8 and 9 were
units of cymarose and one terminal glucose in 4 and two established as shown in Fig. 1.

August 2002
1033
Table 2. 1H-NMR Spectral Data of 2—9 in Pyridine-d5
No.
2
3
4
5
6
7
8
9
Aglycone
6
6.64 d (10.3)a)
5.92 d (10.3)
3.27 br d (10.4)
5.97 br d (8.8)
6.62 d (10.2)
5.91 d (10.2)
3.27 br d (10.5)
5.96 br d (8.8)
6.62 d (10.4)
5.91 d (10.4)
3.24 br d (10.5)
5.94 br d (9.0)
6.62 d (10.4)
5.91 d (10.4)
3.24 br d (10.5)
5.93 br d (8.8)
6.62 d (10.3)
5.91 d (10.3)
3.24 br d (10.7)
5.94 br d (8.8)
6.62 d (10.3)
5.91 d (10.3)
3.24 br d (10.7)
5.94 br d (8.8)
6.61 d (10.5)
5.90 d (10.5)
3.22 br d (10.7)
5.94 br d (8.8)
6.62 d (10.5)
5.91 d (10.5)
7
9
3.23 br d (10.2)
5.94 br d (8.6)
12
17
18
19
21
2.92 dd (10.2, 6.3) 2.92 dd (10.0, 6.6) 2.92 dd (10.0, 6.6) 2.91 dd (10.0, 6.6) 2.91 dd (10.5, 6.6) 2.91 dd (10.3, 6.6) 2.92 dd (10.0, 6.6) 2.92 dd (10.0, 6.8)
1.68 s
0.92 s
2.31 s
1.67 s
0.92 s
2.31 s
1.67 s
0.90 s
2.34 s
1.66 s
0.90 s
2.30 s
1.67 s
0.89 s
2.31 s
1.67 s
0.89 s
2.31 s
1.66 s
0.89 s
2.30 s
1.67 s
0.90 s
2.31 s
Sugar
Anomeric 4.98 br d (9.0)
5.01 br d (9.0)
4.88 d (7.8)
5.02 br d (9.3)
4.98 br d (9.5)
4.90 d (7.8)
5.03 br d (9.5)
4.98 br d (9.0)
4.73 d (7.6)
5.27 br d (9.3)
4.98 br d (9.3)
4.79 d (7.6)
5.10 d (7.8)
5.25 br d (9.3)
5.07 d (7.8)
5.25 br d (9.4)
5.14 d (7.8)
5.04 d (8.0)
5.01 br d (9.3)
4.71 d (7.6)
3.86 s
5.03 br d (9.8)
4.98 br d (9.0)
4.66 d (7.8)
4.97 br d (9.2)
4.71 d (7.6)
OMe
6-Me
3.35 s
3.42 s
3.50 s
3.49 s
3.87 s
3.96 s
3.51 s
3.91 s
3.88 s
3.50 s
3.52 s
3.53 s
3.50 s
3.50 s
3.49 s
1.38 d (6.1)
1.46 d (5.8)
1.56 d (5.8)
1.17 d (5.8)
1.55 d (5.9)
1.54 d (5.9)
1.18 d (5.9)
1.56 d (6.1)
1.49 d (6.1)
1.17 d (6.1)
1.71 d (6.1)
1.50 d (6.1)
1.18 d (6.1)
1.64 d (5.8)
1.52 d (6.1)
1.17 d (5.9)
1.64 d (5.8)
1.51 d (6.1)
1.17 d (5.9)
a) J values in Hz are given in parentheses.
ford compounds 2 (24 mg), 4 (18 mg) and 7 (185 mg).
We isolated a 17-epimer having an a-oriented side chain
of cynaphylloside G (8), that is, 12(R)-O-cinnamoyloxy-
3b,5b-dihydroxy-8,14-seco-17a-pregn-6-ene-8,14,20-trione.
However, this compound might be converted from 8 by keto–
enol interconversion of the side chain during the process of
extraction and isolation. There have been only two reports
of 8,14-seco-pregnane type of compounds from plant
sources.3,6)
The Et2O extract (59 g) was subjected to a column of silica gel using a
gradient system (EtOAc to 80% EtOAc in MeOH) affording six fractions.
Fraction 2 (1.6 g) was chromatographed on RP-18 columns (system VII),
followed by HPLC-ODS (system VIII) to provide compounds 5 (58 mg) and
6 (380 mg). Fraction 4 (4.0 g) was further purified by RP-18 (system IX),
HPLC-ODS (system VII) and HPLC-Diol (system X) to afford compounds
3 (18 mg) and 10 (45 mg).
Cynaphyllogenin (1): White amorphous powder, [a]D21 Ϫ115.8° (MeOH,
cϭ0.61), 1H- and 13C-NMR: Table 1, HR-FAB-MS (negative mode) m/z:
507.2379 (Calcd for C30H35O7, [MϪH]Ϫ 507.2382).
Cynaphylloside A (2): White amorphous powder, [a]D21 Ϫ63.9° (MeOH,
cϭ0.81), 1H-NMR: Table 2, 13C-NMR: Tables 3 and 4, HR-FAB-MS (nega-
tive mode) m/z: 651.3163 (Calcd for C37H47O10, [MϪH]Ϫ: 651.3169).
Cynaphylloside B (3): White amorphous powder, [a]D21 Ϫ63.8° (MeOH,
cϭ0.89), 1H-NMR: Table 2, 13C-NMR: Tables 3 and 4, HR-FAB-MS (nega-
tive mode) m/z: 813.3689 (Calcd for C43H57O15, [MϪH]Ϫ: 813.3697).
Cynaphylloside C (4): White amorphous powder, [a]D21 Ϫ42.4° (MeOH,
cϭ0.96), 1H-NMR: Table 2, 13C-NMR: Tables 3 and 4, HR-FAB-MS (nega-
tive mode) m/z: 957.4480 (Calcd for C50H69O18, [MϪH]Ϫ: 957.4483).
Cynaphylloside D (5): White amorphous powder, [a]D21 Ϫ38.2° (MeOH,
cϭ1.10), 1H-NMR: Table 2, 13C-NMR: Tables 3 and 4, HR-FAB-MS (nega-
tive mode) m/z: 955.4687 (Calcd for C51H71O17, [MϪH]Ϫ: 955.4691).
Cynaphylloside E (6): White amorphous powder, [a]D21 Ϫ61.2° (MeOH,
cϭ0.50), 1H-NMR: Table 2, 13C-NMR: Tables 3 and 4, HR-FAB-MS (nega-
tive mode) m/z: 941.4550 (Calcd for C50H69O17, [MϪH]Ϫ: 941.4534).
Cynaphylloside F (7): White amorphous powder, [a]D21 Ϫ53.9° (MeOH,
cϭ0.59), 1H-NMR: Table 2, 13C-NMR: Tables 3 and 4, HR-FAB-MS (nega-
tive mode) m/z: 1117.5243 (Calcd for C57H81O22, [MϪH]Ϫ: 1117.5219).
Cynaphylloside G (8): White amorphous powder, [a]D21 Ϫ52.6° (MeOH,
cϭ0.91), 1H-NMR: Table 2, 13C-NMR: Tables 3 and 4, HR-FAB-MS (nega-
tive mode) m/z: 1103.5058 (Calcd for C56H79O22, [MϪH]Ϫ: 1103.5062).
Cynaphylloside H (9): White amorphous powder, [a]D21 Ϫ56.1° (MeOH,
Experimental
General Procedure NMR spectra were recorded in C5D5N using a
JEOL JNM A-400 spectrometer (400 MHz for H-NMR and 100 MHz for
1
13C-NMR) with tetramethylsilane (TMS) as internal standard. MS were
recorded on a JEOL JMS-SX 102 spectrometer. Optical rotations were mea-
sured with a Union PM-1 digital polarimeter. Preparative HPLC was carried
out on columns of ODS (150ϫ20 mm i.d., YMC), Polyamine II (250ϫ20
mm i.d., YMC) and Diol (300ϫ8 mm i.d., YMC) with a Tosoh reflection
index (RI-8) detector. Medium pressure liquid chromatography (MPLC) was
carried out on a column of Polyamide C-200. For CC, silica gel G 60
(Merck), YMC-gel ODS (50 mm, YMC) and a highly porous copolymer of
styrene and divinylbenzene (Mitsubishi Chem. Ind. Co., Ltd.) were used.
The solvent systems were: (I) 60% MeOH, (II) EtOAc–MeOH (9 : 1), (III)
70% MeOH, (IV) 65% MeOH, (V) 90% MeCN, (VI) 92% MeCN, (VII)
50% MeCN, (VIII) 55% MeCN, (IX) 50% MeCN and (X) MeCN. The spray
reagent used for TLC was 10% H2S4O in 50% EtOH. Acid hydrolysis of
glycosides followed by identification of the resulting monosaccharides in-
cluding absolute configuration was carried out as previously described.5)
Plant Material The aerial parts of Cynanchum aphyllum were collected
in November 1998 from Ambalavao, Madagascar and identified by Dr. Ar-
mand Rakotozafy, Institut Malgache de Recherches Appliquees, Madagas-
car. A voucher specimen is kept in the Herbarium of the Institute of Pharma-
ceutical Sciences, Faculty of Medicine, Hiroshima University, Japan.
1
cϭ0.71); H-NMR: Table 2, 13C-NMR: Tables 3 and 4, HR-FAB-MS m/z:
1265.5580 (Calcd for C62H89O27, [MϪH]Ϫ: 1265.5591).
Extraction and Isolation The dried aerial parts (800 g) of C. aphyllum
were extracted with hot methanol. After removal of the solvent by evapora-
tion, the residue (166 g) was extracted with n-hexane and Et2O, successively.
The aqueous layer was subjected to a column of highly porous copolymer of
styrene and divinylbenzene, and successively eluted with H2O, 50% MeOH,
MeOH and acetone. The fraction eluted with MeOH (10.5 g) was subjected
to a column of RP-18 (system I) affording eight fractions. Fraction 5 (2.8 g)
was chromatographed on a column of silica gel (system II), followed by
HPLC-ODS (systems III and IV) to provide compounds 1 (18 mg), 8 (1.2 g)
and 9 (43 mg). Fraction 6 (2.3 g) was subjected to a column of MPLC-
Polyamide (system V), followed by HPLC-Polyamine II (system VI) to af-
Partial Acid Hydrolysis of 6 A solution of 6 (45.0 mg) in 2.0 ml of 1,4-
dioxane and 1.0 ml of 0.2 N H2SO4 was heated at 50 °C for 1 h. After 3.0 ml
of H2O was added the mixture was extracted with EtOAc. The EtOAc was
evaporated, followed by HPLC-ODS (60% MeOH) to provide 1 (14.0 mg)
and 2 (4.5 mg). The structures were confirmed by 1H- and 13C-NMR spectral
data.
Enzymatic Hydrolysis of 7—9 Cynaphyllosides F (7, 30 mg), G (8,
30 mg) and H (9, 15 mg) were dissolved in 0.5 ml of MeOH. A solution of
crude hesperidinase (100 mg in 20 ml of H2O) was added. After stirring at
37 °C for 1 week, the mixtures were extracted with EtOAc. The EtOAc was

1034
Vol. 50, No. 8
Table 3. 13C-NMR Spectral Data of Aglycone Moieties of 2—9 in Pyri- Table 4. 13C-NMR Spectral Data of Sugar Moieties of 2—9 in Pyridine-d5
dine-d5
No.
2
3
4
5
6
7
8
9
No.
2
3
4
5
6
7
8
9
Cym
97.6
35.6
78.6
73.9
71.0
18.8
58.0
Cym
97.5
36.4
77.9
82.8
69.4
18.5
58.5
Glc
Cym
Cym
Cym
97.5
36.6
77.8
82.8
69.1
18.3a)
Cym
Cym
97.5
Cym
97.5
36.6
77.8
82.8
69.1
18.2a)
58.8
Dig
1
2
3
4
5
6
7
8
9
26.0
26.7
74.1
36.5
74.2
25.9
26.7
74.1
36.5
74.2
25.9
26.7
74.0
36.4
74.2
25.9
26.7
74.0
36.4
74.2
25.9
26.7
74.0
36.5
74.2
25.9
26.7
74.0
36.4
74.1
25.9
26.7
74.0
36.4
74.2
25.9
26.7
74.0
36.4
74.1
1
97.5
97.5
97.4
2
36.4a)
77.7b)
82.8c)
69.0d)
18.3e)
36.6a)
77.7b)
82.8c)
69.0d)
18.3e)
36.6a)
77.7b)
82.8c)
69.0d)
18.2e)
36.7
3
4
77.8
82.8a)
69.1b)
18.3c)
5
154.1 154.1 154.1 154.1 154.1 154.0 154.1 154.1
127.0 127.0 127.0 127.0 127.0 127.0 127.0 127.0
201.8 201.8 201.8 201.8 201.8 201.8 201.8 201.8
48.0
45.8
26.3
71.3
57.5
6
OMe
58.7f ) 58.8f ) 58.8
58.7f ) 58.8
Cym
Cym
Dig
Cym
Dig
47.9
45.7
26.2
71.3
57.5
48.0
45.8
26.2
71.3
57.5
47.9
45.7
26.3
71.3
57.4
47.9
45.7
26.2
71.2
57.4
47.9
45.7
26.2
71.2
57.4
47.9
45.7
26.2
71.3
57.4
47.9
45.7
26.2
71.2
57.4
1
106.5
75.4
78.4
71.8
78.4
63.0
100.3
36.7a)
77.9b)
83.0c)
69.3d)
18.6e)
100.3
36.9a)
78.1b)
83.0c)
69.3d)
18.5e)
100.4
39.1
67.8
83.4
68.8
18.5a)
100.3
36.9a)
78.0b)
82.9c)
69.2d)
18.4e)
58.9f )
100.4
39.1
100.4
39.1
10
2
11
12
13
14
15
16
17
18
19
20
21
3
4
67.7
67.7
83.4a)
68.7b)
18.6c)
83.4b)
68.7
5
215.7 215.7 215.7 215.7 215.7 215.7 215.7 215.7
6
18.4a)
35.3
20.3
59.2
17.1
17.8
35.2
20.3
59.2
17.1
17.8
35.3
20.3
59.2
17.1
17.8
35.2
20.3
59.2
17.1
17.8
35.3
20.2
59.2
17.1
17.8
35.2
20.2
59.2
17.1
17.8
35.2
20.2
59.2
17.1
17.8
35.2
20.2
59.2
17.1
17.8
OMe
58.8f ) 58.8f )
Glc
The
106.2
75.0
87.8
75.8
72.7
18.5e)
60.9
The
105.8
74.8
87.8
75.8
72.7
18.5a)
60.9
The
105.9
74.7
85.8
83.0
71.9
18.6e)
60.6
The
105.6
74.5
The
1
106.5
75.4
78.4g)
71.8
78.3g)
63.0
105.5
74.5c)
86.0
83.1b)
71.5d)
18.6a)
60.6
2
3
4
85.7
207.1 207.1 207.1 207.1 207.1 207.1 207.1 207.1
31.1 31.1 31.1 31.1 31.1 31.1 31.1 31.1
82.9a)
71.9
5
1Ј
2Ј
3Ј
4Ј
167.0 167.0 167.0 167.0 167.0 166.9 167.0 167.0
118.2 118.3 118.2 118.2 118.2 118.2 118.2 118.2
146.0 146.0 146.0 146.0 146.0 146.0 146.0 146.0
134.8 134.8 134.8 134.7 134.7 134.7 134.7 134.7
6
18.5c)
60.6
OMe
Glc
104.7
75.7
Glc
Glc
1
2
3
4
5
6
104.7
104.5
74.8c)
76.4e)
81.6
76.8e)
62.4f )
Glc
5Ј,9Ј 128.7 128.7 128.7 128.7 128.7 128.6 128.7 128.6
6Ј,8Ј 129.3 129.3 129.3 129.3 129.3 129.3 129.3 129.3
7Ј
75.8
78.5g)
78.6d)
71.9
130.8 130.8 130.8 130.8 130.8 130.8 130.8 130.8
71.9
78.1g)
63.0
78.2d)
63.0
evaporated to provide 5 (23.0 mg) from 7, 6 (21.0 mg) from 8 and 6 (8.0 mg)
from 9. The structures were confirmed by 1H- and 13C-NMR spectral data.
1
2
3
4
5
6
104.9
75.3
78.4g)
71.8d)
78.2g)
62.3f )
Acknowledgements We would like to thank the Research Center for
Molecular Medicine, Hiroshima University for the use of its NMR facilities.
Also, we are grateful to Dr. Rakotozafy Armand, IMRA Madagascar, for
help in collection and identification of the plant. This study was financially
supported by a Grant-in-Aid for the Monbusho International Scientific Re-
search Program (No. 09041185, 1997—1998) from the Ministry of Educa-
tion, Culture, Sports, Science and Technology, Japan.
a—g) Assignments may be interchanged in each vertical column.
4) Lin L.-J., Lin L.-Z., Gil R. R., Cordell G. A., Ramesh M., Srilatha B.,
Reddy B., Rao A. V. A. A., Phytochemistry, 35, 1549—1553 (1994).
5) Huan V. D., Ohtani K., Kasai R., Yamasaki K., Tuu N. V., Chem.
Pharm. Bull., 49, 453—460 (2001).
6) Mu.-Z., Shen Y.-M., Zhou Q.-L., Wang S.-Q., Wu B., Zheng Q.-T.,
Planta Med., 58, 200—204 (1992).
References
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(1989).
2) Deepak D., Srivastav S., Khare A., Fortschr. Chem. Org. Naturstoffe,
71, 169—325 (1997).
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