Four new triterpenes from Ilex cornuta Lindley

Article abstract of DOI:10.1139/cjc-2012-0411

Two new lupane-type triterpenes, 24β-hydroxylupenone (1) and 3β-hydroxy-lup-20(29)-en-24-al (2), and two new ursane-type triterpenes, 28-formyloxy-3β-hydroxy-urs-12-ene (3) and 28-formyloxy-3β-acetoxy- urs-12-ene (4), together with 26 known triterpenes (5

Full text of DOI:10.1139/cjc-2012-0411

382
ARTICLE
Four new triterpenes from Ilex cornuta Lindley
Seung Young Lee, Ho Kyung Kim, and Kang Ro Lee
Abstract: Two new lupane-type triterpenes, 24␤-hydroxylupenone (1) and 3␤-hydroxy-lup-20(29)-en-24-al (2), and two new
ursane-type triterpenes, 28-formyloxy–3␤-hydroxy-urs-12-ene (3) and 28-formyloxy-3␤-acetoxy-urs-12-ene (4), together with 26
known triterpenes (5–30) were isolated from the leaves and trunk of Ilex cornuta Lindl. The structures of the new compounds were
determined by spectroscopic methods, including 1D, 2D NMR (COSY, HMQC, HMBC, and NOESY), and HRMS data.
Key words: Ilex cornuta, Aquifoliaceae, triterpene.
Résumé : L'extraction des feuilles et du tronc de Ilex cornuta Lindl. a permis d'isoler deux nouveaux triterpènes de la famille du
lupane, la 24␤-hydroxylupénone (1) et le 3␤-hydroxylup-20(29)-én-24-al (2), deux nouveaux triterpènes de la famille de l'ursane,
le 28-formyloxy-3-␤-hydroxyurs-12-ène (3) et le 28-formyloxy-3-␤-acétoxyurs-12-ène (4) ainsi que vingt-six triterpèrenes connus
(5–30). Les structures des nouveaux composés ont été déterminées par des méthodes spectroscopiques, dont des données de RMN
1D et 2D [spectroscopie de corrélation (“COSY”); corrélation de liens multiples hétéronucléaires (“HMBC”); corrélation multi-
quantique hétéronucléaire (“HMQC”); spectroscopie de l'effet Overhauser nucléaire (“NOESY”)] et des données de spectrométrie
de masse a` haute résolution (SM-HR). [Traduit par la Rédaction]
Mots-clés : Ilex cornuta, Aquifoliacée, triterpène.
0.87, 0.96, 1.04, and 1.26 (each 3H, s), a isopropenyl group at
Introduction
␦H 1.68 (3H, s), 4.58 (1H, s), and 4.69 (1H, s), and a hydroxymethyl
Ilex cornuta Lindl. (Aquifoliaceae) is an evergreen shrub widely
distributed in Korea and China, and is known as Chinese holly.1
This indigenous plant has been used in Chinese folk medicine for
treating dizziness and hypertension.2 Further, an aqueous extract
from its leaves has been used traditionally as a contraceptive,
cardiovascular system protection, and antibacterial agent.3
Previous phytochemical investigations of I. cornuta have shown
that its leaves and trunk are a rich source of triterpenoids and
flavonoids.3–7 Some of these isolates have been reported to exhibit
various pharmacological effects, such as an increase in coronary
blood flow and antihematoblastic coagulation activity.5,8
In our continuing search for novel secondary metabolites from
Korean medicinal plants, we have investigated the constituents
from the leaves and trunk of I. cornuta. Column chromatographic
separation of the MeOH extract led to isolation of two new lupane-
type triterpenes (1 and 2) and two new ursane-type triterpenes (3
and 4) (Fig. 1), together with 26 known compounds (5–30). The
structures of these new compounds were elucidated by spectro-
scopic methods, including 1D and 2D NMR.
group at ␦H 3.42 (1H, d, J = 11.0 Hz), and 3.97 (1H, d, J = 11.0 Hz). In
the 13C NMR spectrum (Table 2), 30 carbon signals appeared, in-
cluding six methyl carbons at ␦C 14.7, 15.8, 17.2, 18.2, 19.5, and 22.3,
a hydroxymethyl carbon at ␦C 66.0, one ketone carbon at ␦C 221.7,
two olefinic carbons at ␦C 109.6, and 151.1, including 10 methylene,
five methine, and five quaternary carbons. These data implied
that 1 could be a lupane-type triterpene.9 Comparison of the NMR
data for 1 with those of lupenone (5),10 indicated that compound 1
was an analogue of 5. The main difference was an additional
hydroxyl group at C-24, indicating the presence of a hy-
droxymethyl group in 1 instead of a methyl group in 5. The HMBC
spectrum showed correlations from H-24 to C-3, C-4, and C-23,
supporting the presence of the hydroxyl group at C-24 in the
structure (Fig. 2). The relative configuration of 1 was supposed to
be identical to that of 5 based on the J value and chemical shifts in
the 1H NMR spectrum. The hydroxymethyl group at C-24 was de-
termined to be ␤-form, as the NOESY experiment showed a corre-
lation between H-24 (␦H 3.42, and 3.97) and Me-25 (␦H 0.87) (Fig. 3).
On the basis of the NMR data and the spectroscopic data of lupane-
type triterpnenes isolated from this source,6 the structure of 1 was
determined to be 24␤-hydroxylupenone. The oxidation of lup-
20(29)-en-3␤,24-diol (11) by chromium trioxide produced 24␤-
hydroxylupenone, which was identical to the isolated compound
(1) based on NMR spectral data.11,12
Here, we report the isolation of chemical constituents and
structural elucidation of compounds 1–30.
Results and discussion
The 80% MeOH extract of the leaves and trunk of I. cornuta was
subjected to repeated silica gel column chromatography to afford
four new triterpenes (1–4), together with 26 known compounds
(5–30). The structures of the new compounds were established by
spectroscopic and chemical means.
Compound 1 was obtained as a colorless gum whose molecular
formula was determined to be C30H48O2 from the [M + H]+ peak at
m/z 441.3731 (calculated for C30H49O2: 441.3733) in the high reso-
lution fast atom bombardment HRFAB-MS. The 1H NMR spectrum
of 1 (Table 1) displayed signals for five methyl groups at ␦H 0.79,
Compound 2 was obtained as a colorless gum whose molecular
formula was determined to be C30H48O2 from the [M + Na]+ peak at
m/z 463.3553 (calculated for C30H48NaO2: 463.3552) in the HRFAB-
MS. The proton and carbon signals in the 1H and 13C NMR spectra
(Table 1 and 2) of 2 were very similar to those of lupeol (6).10 The
only difference was the appearance of the formyl group signals (␦H
9.76; ␦C 206.9), as well as the disappearance of the methyl carbon
and proton signals (␦H 0.94; ␦C 15.4) in 2.13 The position of the
Received 24 October 2012. Accepted 25 November 2012.
S.Y. Lee, H.K. Kim, and K.R. Lee. Natural Products Laboratory, School of Pharmacy, Sungkyunkwan University, 300 Chonchon-dong, Jangan-ku, Suwon 440-746, Korea.
Corresponding author: Kang Ro Lee (e-mail: krlee@skku.ac.kr).
Can. J. Chem. 91: 382–386 (2013) dx.doi.org/10.1139/cjc-2012-0411
Published at www.nrcresearchpress.com/cjc on 28 November 2012.

Lee et al.
383
Fig. 1. Chemical structures of compounds 1–6, 23, 25, 3a, and 4a.
29
30
20
H
H 21
29
20
19
30
21
H 19
H
12
12
26
22
28
17
11
13
22
17
11
9
18
18
14
15
26
25
13
25
10
1
28 OR2
9
14
1
16
2
16
2
10
8
15
27
8
H
H
H
5
3
3
5
27
4
4
7
7
HO
O
R1O
6
H
6
H
H
24
R3
R 24
23
23
1 R = CH2OH
5 R = CH3
3 R1 = H, R2 = CHO
2 R = CHO
6 R = CH3
4 R1 = COCH3, R2 = CHO
3a = 4a = 23 R1 = R2 = H
25 R1 = COCH3, R2 = H
1
Table 1. H NMR data of compounds 1–4 (CDCl3, 500 MHz, ␦ in ppm, J in Hz).
Protone
1
2
3
4
1
2
1.53, m, 1.90, m
2.40, m, 2.60, m
1.40, m
1.84, m
1.01, m, 1.65, m
1.62, m
1.02, m, 1.65, m
1.65, m
3␣
5
3.16, dd (11.0, 5.5)
0.99, m
3.21, dd (11.0, 5.5)
0.72, m
4.50, dd (11.0, 5.5)
0.85, m
1.58, m
6
7
9
1.40, m, 1.50, m
1.43, m
1.40, m
1.46, m
1.42, m
1.34, m
1.42, m, 1.57, m
1.40, m, 1.57, m
1.53, m
1.40, m, 1.55, m
1.38, m, 1.57, m
1.53, m
11
12
13
15
16
18
19
20
21
22
23
24
1.40, m
1.69, m
1.70, m
1.70, m
1.36, m, 1.53, m
1.38, m
2.38, m
1.48, m
1.70, m
1.78, m
1.70, m
1.38, m, 1.54, m
1.36, m
2.38, m
1.93, m
5.16, br s
1.95, m
5.16, br s
1.04, m, 1.18, m
1.12, m, 1.23, m
1.42, m
1.42, m
1.00, m
1.27, m, 1.48, m
1.40, m, 1.60, m
1.00, s
1.05, m, 1.17, m
1.12, m, 1.23, m
1.43, m
1.45, m
0.95, m
1.25, m, 1.48, m
1.38, m, 1.60, m
0.99, s
1.35, m, 1.90, m
1.20, m, 1.40, m
1.26, s
3.42, d (11.0)
3.97, d (11.0)
0.87, s
1.04, s
0.96, s
0.79, s
1.30, m, 1.90, m
1.20, m, 1.40, m
1.26, s
9.76, s
0.79, s
0.88, s
25
26
27
28
0.77, s
1.04, s
0.96, s
0.79, s
0.95, s
1.03, s
1.12, s
3.74, d (11.0)
4.15, d (11.0)
0.84, d (6.5)
0.93, d (6.5)
8.08, s
0.95, s
1.03, s
1.13, s
3.74, d (11.0)
4.15, d (11.0)
0.81, d (6.5)
0.94, d (6.5)
8.08, s
29
30
CHO
COCH3
4.58, s, 4.69, s
1.68, s
4.57, s, 4.69, s
1.68, s
2.04, s
Note: Assignments were based on HMQC, and HMBC experiments.
formyl group in 2 was confirmed by HMBC, which showed long-
range correlations between the formyl proton (␦H 9.76) and the
C-3 (␦C 78.0); the Me-23 (␦H 1.26) and the C-3 (␦C 78.0); C-4 (␦C 52.7)
and the formyl carbonyl carbon (␦C 206.9); as well as the H-3 (␦H
3.16) and the formyl carbonyl carbon (␦C 206.9) (Fig. 2). The above
data showed that the position of the formyl group was located at
C-24. The coupling constants of H-3 (␦H 3.16, dd, J = 11.0, 5.5 Hz)
0.84 (d, J = 6.5 Hz), 0.93 (3H, d, J = 6.5 Hz), 0.95 (s), 1.00 (s), 1.03 (s),
and 1.12 (s); an oxygenated methine signal at ␦H 3.21 (1H, dd, J =
11.0, 5.5 Hz); and a olefinic signal at ␦H 5.16 (1H, br s), as well as a
hydroxymethyl group at ␦H 3.74 (1H, d, J = 11.0 Hz), and 4.15 (1H, d,
J = 11.0 Hz) (Table 1). The 13C NMR spectrum indicated 30 carbon
resonances, which were classified by HMQC experiment as
olefinic carbon signals at ␦C 125.8 and 138.0, an oxygenated car-
bons at ␦C 79.0, a hydroxymethyl carbon at ␦C 70.8, seven methyl,
nine methylene, five methine, and five quaternary carbons
1
observed in the H NMR spectrum indicated ␤-orientation of the
OH group at C-3 (dd, J = 11.0, 5.5 Hz).10,14 The ␤-orientation of the
formyl group was determined from correlations between the for-
myl proton (␦H 9.76)/Me-25 (␦H 0.77) and H␣-3 (␦H 3.16)/Me-5 (␦H
0.99) in the NOESY spectrum (Fig. 3). Thus, the structure of 2 was
determined to be 3␤-hydroxy-lup-20(29)-en-24-al.
1
(Table 2). The H and 13C NMR spectra of 3 were very similar to
those of 3␤,28-dihydroxy-urs-12-ene (23),15 except for the presence
of formyl group signals [␦H 8.08 (1H, s); ␦C 161.3],16,17 which were
located at C-28 based on the HMBC correlation between the H-28
(␦H 3.74, and 4.15) and the formyl carbonyl carbon (␦C 161.3) (Fig. 2).
The stereochemistry of 3 was established based on the NOESY
correlation between H-3 and H-5, and no correlation between H-3
and H-25 indicated that the OH group at C-3 was in ␤-form (Fig. 3).
Compound 3 was obtained as a colorless gum whose molecular
formula was determined to be C31H50O3 from the [M]+ peak at m/z
470.3761 (calculated for C31H50O3: 470.3760) in the HRFAB-MS. The
1H NMR spectrum of 3 showed seven methyl signals at ␦H 0.79 (s),
Published by NRC Research Press

384
Can. J. Chem. Vol. 91, 2013
Table 2. 13C NMR data of compounds 1–4 (CDCl3, 125 MHz, ␦ in ppm).
Experimental
Carbon
1
2
3
4
General experimental procedures
Optical rotations were measured in MeOH using a Jasco P-1020
polarimeter. IR spectra were recorded on a Bruker IFS-66/S FT-IR
spectrometer. HRFAB mass spectra were obtained on a JEOL
1
2
3
4
5
6
7
8
39.5
34.5
40.0
28.2
78.0
52.7
56.3
21.3
34.7
40.6
49.3
36.0
22.6
25.3
38.4
42.9
27.4
35.5
43.0
48.2
47.9
150.8
29.8
40.1
19.3
206.9
15.0
16.0
14.4
18.0
109.4
19.3
39.2
38.6
23.8
81.1
28.1
79.0
38.9
55.2
18.3
32.7
40.2
47.6
36.9
23.3
125.8
138.0
42.1
25.9
23.2
38.0
54.2
39.7
39.9
30.4
35.6
28.2
15.5
15.6
17.2
221.7
50.9
55.5
19.4
33.8
40.9
49.8
36.8
22.1
25.4
38.4
43.1
27.6
35.7
43.2
48.5
48.1
151.1
30.1
40.2
22.3
66.0
17.2
15.8
14.7
18.2
109.6
19.5
1
JMS700 mass spectrometer. NMR spectra, including H-1H COSY,
37.9
55.5
18.4
32.9
40.2
47.8
37.0
23.6
126.0
138.3
42.2
26.2
23.5
38.2
54.4
39.5
40.2
30.6
35.8
28.2
15.9
16.2
17.5
HSQC, HMBC, and NOESY experiments, were recorded on a Varian
UNITY INOVA 500 NMR spectrometer operating at 500 MHz (1H)
and 125 MHz (13C) with chemical shifts given in ppm (␦). Prepara-
tive HPLC was conducted using a Gilson 306 pump with Shodex
refractive index detector and Apollo Silica 5 ␮ column (250 ×
10 mm). Silica gel 60 (Merck, 70–230 mesh and 230–400 mesh) and
RP-C18 silica gel (Merck, 230–400 mesh) were used for column
chromatography. Merck precoated Silica gel F254 plates and RP–18
F254s plates were used for TLC. Spots were detected on TLC under
UV light or by heating after spraying with 10% H2SO4 in
C2H5OH (v/v).
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
CHO
COCH3
COCH3
Plant material
The leaves and trunk from I. cornuta (5.0 kg) were collected on
Jeju Island, Korea in March 2011, and the plant was identified by
one of the authors (K.R. Lee). A voucher specimen (SKKU-NPL 1108)
has been deposited at the herbarium in the School of Pharmacy,
Sungkyunkwan University, Suwon, Korea.
Extraction and isolation
The leaves from I. cornuta (5.0 kg) were extracted with 80% aque-
ous MeOH under reflux and filtered. The resulting MeOH extracts
(550 g) were suspended in distilled water (800 mL × 3) and then
successively partitioned with n-hexane, CH2Cl2, EtOAc, and
n-BuOH, yielding 20, 68, 28, and 100 g, respectively. The n-hexane
soluble fraction (20 g) was chromatographed on a silica gel (230–
400 mesh, 500 g) column eluted with n-hexane-EtOAc (15:1 – 1:1,
gradient system) to yield 10 fractions (H1–H10). Fraction H1 (50 mg)
was purified by preparative HPLC (n-hexane–EtOAc, 50:1) to give
compounds 15 (25 mg) and 16 (6 mg). Fraction H2 (1.5 g) was
chromatographed on a silica gel (230–400 mesh, 40 g) column
eluted with n-hexane–EtOAc (40:1) to give five subfractions (H21–
H25). Subfraction H21 (70 mg) was purified by preparative HPLC
(n-hexane–EtOAc, 30:1) to give 9 (7 mg). Subfraction H22 (200 mg)
was separated by preparative HPLC (n-hexane–EtOAc, 85:1) to give
8 (7 mg) and 17 (10 mg). Subfraction H23 (40 mg) was separated by
preparative HPLC (n-hexane–EtOAc, 30:1) to obtain 5 (20 mg). Sub-
fraction H24 (250 mg) was purified by preparative HPLC (n-hexane–
EtOAc, 50:1) to give 18 (5 mg) and 19 (23 mg). Subfraction H25
(40 mg) was separated by preparative HPLC (n-hexane–EtOAc, 30:1)
to afford 4 (5 mg). Fraction H4 (1.2 g) was chromatographed on a
silica gel (230–400 mesh, 30 g) column eluted with n-hexane–
EtOAc (40:1) to give three subfractions (H41–H43). Subfraction H42
(50 mg) was purified by preparative HPLC (n-hexane–EtOAc, 15:1)
to give 7 (12 mg). Fraction H5 (5 g) was chromatographed on a silica
gel (230–400 mesh, 100 g) column eluted with n-hexane–EtOAc
(30:1) and purified further by preparative HPLC (n-hexane–EtOAc,
10:1) to give 6 (4 g). Fraction H6 (2 g) was chromatographed on an
RP-C18 silica gel (230–400 mesh, 60 g) column eluted with 100%
MeOH to give three subfractions (H61–H63). Subfraction H61
(200 mg) was purified by preparative HPLC (n-hexane–EtOAc, 10:1)
to give 2 (4 mg), 11 (20 mg), 24 (27 mg), and 26 (4 mg). Subfraction
H62 (100 mg) was purified by preparative HPLC (n-hexane–EtOAc,
5:1) to give 3 (7 mg), 10 (4 mg), and 25 (4 mg). Fraction H8 (5 g) was
chromatographed on a silica gel (230–400 mesh, 120 g) column
eluted with n-hexane–EtOAc (40:1) to give six subfractions (H81–
H86). Subfraction H81 (150 mg) was purified by preparative HPLC
(100% MeOH) to give 1 (4 mg) and 13 (15 mg). Subfraction H83
(120 mg) was separated by preparative HPLC (95% MeOH) to afford
12 (60 mg), 14 (4 mg), and 29 (10 mg). Subfraction H84 (70 mg) was
purified by preparative HPLC (n-hexane–EtOAc, 4:1) to yield 20
23.4
70.8
17.2
21.2
161.3
23.3
70.1
17.5
21.4
161.5
171.2
21.4
Alkaline hydrolysis of 3 afforded 3␤,28-dihydroxy-urs-12-ene (3a),
which was identified by comparing its optical rotation value as
well as H NMR and MS data.24 Therefore, the structure of 3 was
1
determined to be 28-formyloxy-3␤-hydroxy-urs-12-ene.
Compound 4 was obtained as a colorless gum whose molecular
formula was determined to be C33H52O4 from the [M + Na]+ peak at
m/z 535.3762 (calculated for C33H52NaO4: 535.3763) in the HRFAB-
MS. The 1H and 13C NMR spectra of 4 were very similar to those of
3. The only difference was the presence of additional acetyl group
signals [␦H 2.04 (1H, s); ␦C 21.4, and 161.3].18 The position of the
acetyl group was identified by the HMBC spectrum, in which a
long-range correlation was observed between the H-3 (␦H 4.50, dd,
J = 10.5, 5.5 Hz) and a carbonyl carbon (␦C 171.2) (Fig. 2). Alkaline
hydrolysis of 4 yielded 3␤,28-dihydroxy-urs-12-ene (4a), which was
1
identified by comparing its optical rotation value, H NMR and
MS data.15 Hence, the structure of 4 was determined to be
28-formyloxy-3␤-acetoxy-urs-12-ene.
The structures of the known compounds (5–30) were identified
as lupenone (5),10 lupeol (6),10 3-epi-lupeol (7),14 lupeol acetate (8),19
lupenyl formate (9),20 3-O-acetylbetulin (10),21 lup-20(29)-en-3␤,
24-diol (11),22 3␤-hydroxy-lup-20(29)-en-30-al (12),23 3␤-hydroxy-
20-oxo-30-norlupane (13),24 betulone (14),25 ␣-amyrin palmitate
(15),26 ␤-amyrin palmitate (16),27 ␣-amyrin acetate (17),27
11-oxo-␣-amyrin palmitate (18),28 11-oxo-␤-amyrin palmitate (19),29
11-oxo-␣-amyrin (20),30 11-oxo-␤-amyrin (21),30 3␤,28-dihydroxy-
ole-12-ene (22),31 3␤,28-dihydroxy-urs-12-ene (23),15 3␤-acetoxy-28-
hydroxy-ole-2-ene (24),32 3␤-acetoxy-28-hydroxy-urs-12-ene (25),18
3␤-acetoxy-13(28)-epoxy-ole-11-ene (26),33 3␤-hydroxy-13(28)-epoxy-
urs-11-ene (27),34 3␤-hydroxy-11␣-methoxy-urs-12-ene (28),33
28-nor-urs-12-ene-3␤,17␤-diol (29),34 and 17␤-formyloxy-28-nor-urs-
12-en-3␤-ol (30)35 by comparison of 1H and 13C NMR, and MS data with
those reported previously.
Published by NRC Research Press

Lee et al.
385
Fig. 2. Key HMBC (¡) of 1–4.
HO
O
H
OH
O
1
2
O
O
O
O
H
H
O
HO
O
3
4
Fig. 3. Key NOESY ( ) correlations of 1–4.
H
H
H
HO
O
H H
H OH
O
1
2
H
H
O
O
O
H
H
O
O
H
H
HO
O
H
H
3
4
(4 mg), 21 (5 mg), 27 (13 mg), and 30 (13 mg). Fraction H9 (3 g) was
chromatographed on an RP-C18 silica gel (230–400 mesh, 60 g)
column eluted with 100% MeOH and purified by preparative HPLC
(n-hexane–EtOAc, 4:1) to give 22 (5 mg) and 23 (20 mg). Fraction
H10 (1 g) was chromatographed on a silica gel (230–400 mesh, 30 g)
column eluted with n-hexane–EtOAc (15:1) to give four subfrac-
tions (H101–H104). Subfraction H102 (30 mg) was purified by pre-
parative HPLC (n-hexane–EtOAc, 4:1) to give 28 (4 mg).
3␤-Hydroxy-lup-20(29)-en-24-al (2)
Colorless gum; [␣]D25 + 9.0 (c 0.15, CHCl3). IR (KBr) ␯max: 3452,
2944, 1710, 1456, 1380, 1219, 1078, 883, 771, 631 cm–1. H and 13C
NMR (see Tables 1 and 2). HRFAB-MS m/z: 463.3553 [M + Na]+ (cal-
culated for C30H48NaO2, 463.3552).
1
28-Formyloxy-3␤-hydroxy-urs-12-ene (3)
Colorless gum; [␣]D25 + 6.0 (c 0.25, CHCl3). IR (KBr) ␯max: 3419,
2928, 2869, 1722, 1457, 1379, 1185, 1092, 1043, 997, 769, 663 cm–1. 1H
and 13C NMR (see Table 1 and 2). HRFAB-MS m/z: 470.3761 [M]+
(calculated for C31H50O3, 470.3760).
24-Hydroxylupenone (1)
Colorless gum; [␣]D25 + 43.6 (c 0.15, CHCl3). IR (KBr) ␯max: 3454,
2944, 2867, 1702, 1640, 1457, 1381, 1218, 1041, 883, 758 cm–1. 1H and
13C NMR (see Table 1 and 2). HRFAB-MS m/z: 441.3731 [M + H]+
(calculated for C30H49O2, 441.3733).
28-Formyloxy-3␤-acetoxy-urs-12-ene (4)
Colorless gum; [␣]D25 + 13.0 (c 0.20, CHCl3). IR (KBr) ␯max: 3451,
2929, 2869, 1731, 1459, 1372, 1249, 1028, 771, 610 cm−1. 1H and 13
C
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386
Can. J. Chem. Vol. 91, 2013
NMR (see Table 1 and 2). HRFAB-MS m/z: 535.3762 [M + Na]+ (calcu-
lated for C33H52NaO4, 535.3763).
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1a: Colorless gum; [␣]D25 + 15.0 (c 0.07, CHCl3). IR (KBr) ␯
:
max
1
3425 (OH), 1705 (C = O), 1650 (C = C) cm–1. H NMR (CDCl3, 500
MHz) ␦: 0.78 (3H, s, H-24), 0.86 (3H, s, H-25), 0.95 (3H, s, H-27),
1.03 (3H, s, H-26), 1.25 (3H, s, H-23), 1.67 (3H, s, H-30), 2.36 (1H, m,
H-19), 3.42 (1H, d, J = 11.0 Hz, H-24a), 3.96 (1H, d, J = 11.0 Hz,
H-24b), 4.58 (1H, s, H-29a), 4.68 (1H, s, H-29b). 13C NMR (CDCl3,
125 MHz) ␦: 14.6 (C-27), 15.8 (C-26), 17.2 (C-25), 18.2 (C-28), 19.5
(C-30), 19.6 (C-6), 22.0 (C-11), 22.3 (C-23), 25.4 (C-12), 27.6 (C-15),
30.1 (C-21), 33.8 (C-7), 34.6 (C-2), 35.7 (C-16), 36.9 (C-10), 38.4
(C-13), 39.5 (C-1), 40.2 (C-22), 40.9 (C-8), 43.1 (C-14), 43.2 (C-17),
48.2 (C-19), 48.5 (C-18), 49.8 (C-9), 51.0 (C-4), 55.5 (C-5), 66.0 (C-24),
109.6 (C-29), 151.1 (C-20), 221.7 (C-3). FAB-MS m/z: 441 [M + H]+.
Alkaline methanolysis of compounds 3 and 415
Compounds 3 and 4 (each 1.5 mg) were hydrolyzed with 0.5
mol/L KOH in MeOH (1 mL) at room temperature for 1 h. And then,
H2O (3 mL) was added and the mixture was extracted three times
with CHCl3, and the CHCl3 extract was evaporated in vacuo. The
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Vac 12cc cartridge (n-hexane–EtOAc, 5:1) to give 3a (0.8 mg), and 4a
(0.7 mg) (same as 23), which were identified by 1H NMR, MS, and
optical rotation value.
3a (= 4a): Colorless gum; [␣]2D5 + 18.0 (c 0.04, CHCl3). IR (KBr)
1
␯
max: 3350 (OH), 2910 (C-H) cm–1. H NMR (CDCl3, 500 MHz) ␦:
0.79 (3H, s, H-24), 0.81 (3H, d, J = 6.5 Hz, H-29), 0.92 (3H, d, J = 6.5
Hz, H-30), 0.95 (3H, s, H-25), 0.99 (3H, s, H-23), 1.00 (3H, s, H-26),
1.10 (3H, s, H-27), 3.20 (1H, d, J = 10.5 Hz, H-28a), 3.23 (1H, dd, J =
11.5, 5.0 Hz, H-3), 3.55 (1H, d, J = 10.5 Hz, H-28b), 5.14 (1H, br s,
H-12). 13C NMR (CDCl3, 125 MHz) ␦: 15.6 (C-25), 15.7 (C-24), 16.7
(C-26), 17.3 (C-29), 18.3 (C-6), 21.3 (C-30), 23.2 (C-27), 23.3 (C-11),
23.4 (C-16), 26.0 (C-2), 27.2 (C-15), 28.1 (C-23), 30.6 (C-21), 32.8
(C-7), 35.2 (C-22), 36.9 (C-10), 38.0 (C-1), 38.7 (C-4), 39.3 (C-19), 39.4
(C-20), 40.0 (C-8), 42.0 (C-14), 47.6 (C-9), 47.7 (C-17), 54.0 (C-18),
55.2 (C-5), 69.9 (C-28), 79.0 (C-3), 125.0 (C-12), 138.7 (C-13).
HRFAB-MS m/z: 443 [M + H]+.
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Supplementary data are available with the article through the
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Acknowledgements
We thank Dr. Do Kyun Kim, Dr. Eun Jung Bang, and Dr. Jung Ju
Seo at the Korea Basic Science Institute for their aid in obtaining
the NMR and mass spectra. This work was supported by the R & D
Institute, Richwood Pharmaceutical Co., Ltd., Korea.
Published by NRC Research Press