Phytochemistry Vol. 49, No. 7, pp.
2053±2056, 1998
# 1998 Published by Elsevier Science Ltd. All rights reserved
Printed in Great Britain
0031-9422/98/$ - see front matter
PII: S0031-9422(98)00400-2
24-METHYLENE TETRACYCLIC TRITERPENES FROM
POLYALTHIA LANCILIMBA
Y. P. LUE$, Q. MU, H. L. ZHENG% and C. M. LI*
Laboratory of Phytochemistry, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming
650204, P.R. China; %XiShuangbanna Tropical Botanic Garden, Chinese Academy of Sciences, Mengla
666303, P.R. China
(Received in revised form 27 April 1998)
Key Word IndexÐPolyalthia lancilimba; Annonaceae; 24-methylene tetracyclic triterpene;
lanostane.
AbstractÐTwo new 24-methylene tetracyclic triterpenes that possess the lanostane skeleton have been
isolated from the stem barks of Polyalthia lamcilimba. Their structures were determined by spectral analysis
and comparisons with other lanostane type triterpenes described in the literature. The two new components
(1 and 3) were elucidated as 24-methylenelanosta-7,9(11)-dien-3b,15a-diol and 24-methylenelanosta-8-en-
2b,3b,21-triol. # 1998 Published by Elsevier Science Ltd. All rights reserved
INTRODUCTION sence of two ole®nic protons, an exomethylene, two
oxymethines, three secondary and ®ve tertiary
The genus Polyalthia has 120 species. Earlier work
methyls. The 13 C NMR spectrum showed 31 car-
on 14 species of this genus has yielded diterpenes,
bons including 8 methyls, 8 methylenes, 8 methines
triterpenes, nitrogen heterocycles, (zincpolyena-
and 7 quaternary carbons. The HR-EI mass spec-
mine), indolosesquiterpenes, benzylisoquinolines,
trum showed the molecular ion peak at m/z
protoberberines, bisbenzylisoquinolines and apor-
454.3812 (calc. mass 454.3811) corresponding to the
phines. About 14 diterpenes have been isolated
molecular formula C31H50O2. These data suggested
from 6 Polyalthia species [1±3], but only one new
that 1 was a triterpene which had one additional
triterpene, polycarpol, together with some sterols
sp2-methylene [6]. The fragment ion peak in the EI-
(sitosterol, stimasterol, campesterol and cholester-
nol) have so far been reported [4, 5]. Thus few tri- mass spectrum at m/z 327 represented the loss of 9
terpene derivatives have been reported from the carbon atoms, which unit (C9H9+2H) was due to
Annonaceae family. the cleavage of the bond between C-17 and C-20.
For Polyalthia lancilimba collected from Yunnan, Thus compound 1 is a tetracyclic triterpene bearing
China, no chemical studies have been reported to a 9 carbon side chain. The 13 C NMR data corre-
date. In this paper, we wish to report the isolation sponding to these 9 carbons of the side chain are
and structural elucidation of two new 24-methylene almost identical upon comparison with those of
tetracyclic triterpenes. 24-methylenelanosta-7,9(11)dien-3b-ol (2) [7]. This
enabled the 9 carbon skeleton of the side chain of 1
to be identi®ed and assigned.
The co-incidence of 13 C NMR and 1 H NMR
RESULTS AND DISCUSSION
spectral data with those reported in the literature
We have isolated two main components, 1 and 3, suggested that there were striking resemblances
from the ethanolic extract of the stem barks of between 1 and 2, such as two endo-double bonds, a
Polyalthia lancilimba. 3b-OH and other groups in the ring A, B and C
Compound 1, represented 0.8% of the plant besides the side chain. The 13 C NMR signals at d
material. The 1 H NMR spectrum indicated the pre- 121.3, 140.9, 146.2, 116.0 and the 1 H NMR signals
at d 5.82, 5.28 indicated that the two double bonds
were positioned between C-7 and C-8 and between
*Author to whom correspondence should be addressed.
{Present address: Chemistry Department of Yunnan C-9 and C-11. In the COLOC spectrum, two
Normal University, P.R. China. important correlation spots between one ole®nic
2053
2054 Y. P. LUE et al.
carbon and H-19, another ole®nic carbon and H-30 After determination of 15a-OH, the structure of 1
also supported this suggestion. The peaks of one was established as 24-methylenelanosta-7,9(11)dien-
hydrogen at d 3.22 (dd) correspond to H-3. Indeed, 3b,15a-diol. The EI-mass spectral fragmentation
a NOE eect between H-3 and H-28 indicated that pattern also supported the structure.
H-3 was a-oriented and therefore the 3-OH was a Compound 3 is a component with 31 carbons,
b-OH as in 2. These facts established that 1 was a three oxy-groups (d 71.3, 78.3 and 62.5 in the 13 C
24-methylene-7,9(11)-diene of the lanostane type of NMR spectrum, d 4.04, 3.13 and 3.65 in the 1 H
triterpene. NMR spectrum), together with a molecular ion
The dierences between 1 and 2 were in the peak at m/z 472.3925 (calc. mass 472.3916) in the
presence of an extra hydroxyl in the ring system HR-EI-mass spectrum which revealed the formula
(besides 3b-OH) and the spectral data of the vicinal of 3 as C31H52O3. It had 6 unsaturated positions,
protons under the in¯uence of this extra OH. The including two double bonds (d 133.8, 135.3, 106.2,
additional hydroxyl was positioned on ring D (poss- 156.6 in the 13 C NMR spectrum) and 4 rings. These
ible position: 15-OH or 16-OH), because in the EI- facts suggested that 3 possesses a 9 carbon side
mass spectrum, the peak at m/z 273 which included chain. The data in the 13 C NMR and 1 H NMR
only one hydroxyl (3-OH) was due to the cleavage spectra demonstrated a terminal ole®nic bond, 2
of ring D. A close observation of the chemical shifts methyls and a methine which were almost identical
in the 13 C NMR spectra of 1 and 2 of C-14, C-15, by comparison with C-24 to C-27 and C-24 of 1.
C-16 and C-17 showed dierences. However the This indicated the 24-methylene 9 carbon skeleton
chemical shifts of C-20, C-18 and C-30 were identi- of the side chain of 3.
cal (see Table 2). The chemical shifts at d 133.8 and 135.3 of two
In the 1 H±1 H COSY spectrum, the correlation quaternary carbons in 13 C NMR spectrum indi-
between H-20 and a methine proton (H-17 at d cated that 3 had only an endo-ole®nic bond. By
1.68) was determined, then the corresponding car- comparing with the 13 C NMR spectral data of both
bon (C-17) was assigned at d 48.9 in the 1 H±13 C the related compound 4 [8] and lanosterol [9±12],
COSY spectrum. The up®eld chemical shift of the chemical shifts of the carbons of rings B, C, D
C-17 (2.0 ppm) is negligible by comparison with 2. (from C-5 to C-18 and C-30) were similar (see
Thus, we inferred the hydroxyl was not positioned Table 2). These facts showed that compound 3 was
at C-16, but at C-15. This suggestion was supported a lanostane type triterpene having a double bond
by the 1 H±1 H COSY spectrum: H-17 also corre- between C-8 and C-9. On the other hand, the
lated with the protons of a methylene (at d 1.72 cleavage of ring B or C in the EI-mass spectrum
and 1.95). Based on the above analysis, the other supported the position of this double bond [8]. The
hydroxyl has been placed at C-15 and the NOE fragment ion peak at m/z 271 [M±a±H2O]+ rep-
eect between H-15 and H-18 indicated that the resented successive losses of fraction a and water
15-OH was a-oriented. from the molecular ion at m/z 472, together with an
A triterpene from P. lancilimba 2055
ion at m/z 133 [M±a±side chain±Me]+, m/z 161[M± and was regarded as a useful chemotaxonomic mar-
b±H2O±(Me)2]+. ker of Polyalthia. The isolation and identi®cation of
The protons of a methylene substituted by a 1 and 3, which possess a 24-methylenelanostane
hydroxyl did not show a single or doublet peak in structure may be an addition chemotaxonomic
the 1 H NMR spectrum, but two dd were observed. matter.
This indicated that the hydroxyl is not located at
one of the methyls of the carbon rings, but rather
at one of the methyls in the side chain (the possible EXPERIMENTAL
positions were C-26, C-27 and C-21). Also in the M.p. uncorr. UV was obtained on a Shimatsu
13
C NMR spectrum the chemical shift of C-25 (d UV-210A. IR was recorded on a Perkin-Elmer 577.
33.8, the same for 1 and 2) was assigned. In the EI-MS was measured on a Finnigan-4510. NMR
1
H±1 H COSY spectrum, H-25 (d 2.21) showed two were recorded with a Bruker AM-400 in CDCl3,
correlation spots with protons of two methyls with TMS as int. standard. The stems of Polyalthia
whose chemical shifts were at higher ®eld (H-26 at lancilimba were collected from Pingbian of the
d 0.97, H-27 at d 0.98). Thus the ®rst hydroxyl was Yunnan province and its specimen was determined
not positioned at C-26 or C-27, but was located by Mrs Huang Suhua in Yunnan University.
at C-21.
For the two other hydroxyls, a b-OH positioned Extraction and isolation
at C-3 was determined by comparison with the Stem barks of Polyalthia lancilimba (2700 g) were
spectral data of 2 and 4. After H-3 (d 3.13) and C-3 extracted with 95% EtOH and the extract concen-
(d 78.3) were assigned, the 1 H±1 H COSY spectrum trated in vacuo. It then was suspended in H2O and
showed that H-3 was only correlated with one H-2 extracted with CHCl3. The CHCl3 fraction (50 g)
(d 4.04). Therefore the third hydroxyl was posi- was subjected to CC on silica gel eluting with a
tioned at C-2. The NOE eect between H-3 and gradient of petroleum±EtOAc (100:0 to 0:100) and
H-28 supported the b-orientation of the 3-OH. aorded 1 (2130 mg) and 3 (60 mg).
Another NOE eect between H-2 and H-3 indicated 24-Methylenelanosta-7,9(11)-dien-3b,15a-diol (1):
that the 2-OH was also b-oriented. The analysis Yield: 0.8-, white powder m.p. 144±1468,
therefore con®rmed the structure of compound 3 as [a]29 KBr ÿ1
D = + 63.88(c 0.48, C5H5N). IR nmax cm : 3300,
24-methylenelanosta-8-en-2b,3b,21-triol. 2930, 1640, 1460, 1370, 1035, 990, 890. UV lMeOH max
Few triterpenoids have been reported from the nm (log e): 196.5 (3.13), 234 (3.73), 243.5 (3.84),
Annonaceae and only a new triterpene, polycarpol 251.5 (3.67). EIMS, 70 eV, m/z (rel. int.): 454 (100),
(5), has been reported which was isolated from 439 (9), 439 (9), 436 (6), 421 (31), 403 (5), 355 (6),
P. oliveri Engl. and P. suaveolens Engl. et Diels [5, 6] 327 (45), 311 (6), 273 (26), 255 (19), 239 (16), 228
Table 1. 1 H NMR spectral data of compounds 1, 2 and 3$ (d ppm; J, Hz; in CDCl3)
H 1 2 3
H-1 2.01 br 1.38 d, J = 2.8;
2.15 d, J = 2.8
H-2 1.68 m 4.04 m
H-3 3.22 dd, J = 10.9, 5.0 3.24 dd 3.13 d, J = 3.7
H-5 1.07 m 1.80 br t
H-6 2.11 br 1.71 m
H-7 5.82 d, J = 6.9 5.30 d 2.03 br
H-11 5.28 d, J = 6.0 5.46 br 2.06 m
H-12 2.05 br; 2.24 br 1.19 m; 1.57 m
H-15 4.25 dd, J = 9.6, 5.8 1.95 m; 2.10 m
H-16 1.72 m; 1.95 m 1.43 m; 1.56 m
H-17 1.68 m 1.12 m
H-18 0.58 s 0.56 s 0.68 s
H-19 0.91 s 1.00 s 1.33 s
H-20 1.32 m 1.44 s
H-21 0.86 d, J = 6.5 0.96 s 3.69 dd, J = 9.4, 3.3;
3.61 dd, J = 11.0, 3.8
H-22 1.11 m; 1.50 m 1.63 m; 1.77 m
H-23 1.85 m; 2.10 m 1.26 br; 1.95 br
H-25 2.19 hept, J = 6.4 2.21 hept, J = 6.6
H-26 1.00 d, J = 6.9% 1.01 d 0.97 d, J = 6.9%
H-27 0.99 d, J = 6.8% 1.02 d 0.98 d, J = 6.9%
H-28 0.97 s 0.97 s 0.97 s
H-29 0.85 s 0.87 s 0.97 s
H-30 0.95 s 0.87 s 0.84 s
H-241 4.62 s; 4.69 s 4.65 s; 4.71 s 4.64 s; 4.69 s
$Compounds 1 and 3: Bruker AM-400 MHz; compound 2: 360 MHz [7].
%Assignments of H-26 and H-27 may be interchanged.
2056 Y. P. LUE et al.
Table 2. 13
C NMR spectral data of the compounds 1, 2, 3 203.5(3.85); EIMS, 70 eV, m/z (rel. int.): 472 [M]+
and 4 (d ppm, in CDCl3) (73), 457 [M±Me]+ (57), 439 (M±Me±H2O]+ (34),
421 [M±Me±2H2O]+ (25), 315 [M±C9H17O±Me±
C 1 2 3 4
H]+ (17), 297 [315-H2O]+ (11), 282 [297ÿMe]+
C-1 35.8 t 35.7 t 41.3 t 30.2 t (12), 271 (11), 161 (31), 133 (39), 121 (51), 109 (61),
C-2 27.8 t 27.8 t 71.3 t 25.9 t 95 (69), 69 (79). 1 H NMR, 13 C NMR spectra see
C-3 78.9 d 78.9 d 78.3 d 76.7 d
C-4 38.7 s 38.6 s 38.3 s 37.1 s
Tables 1 and 2.
C-5 49.1 d 50.3 d 44.6 d 44.4 d
C-6 23.0 t 22.9 t 17.9 t 18.3 t
C-7 121.3 d 120.1 d 26.4 t 26.2 t
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24-Methylene-lanosta-8-en-2b,3b,21-triol (3): Tetrahedron Lett., 1975, 22, 1787.
Yield: 0.022-, white needles (MeOH), m.p. 152± 12. Lukas, G., Khuong-Huu, F., Bennett, C. R.,
1548, [a]29 KBr
D = + 70.98(c 0.41, MeOH). IR nmax cm :
ÿ1
Buckwalter, B. L. and Wenkert, E., Tetrahdron
MeOH
3400, 2910, 1450, 1365, 1020. UV lmax nm (log e): Lett., 1972, 33, 3515.
