Tetrahedron 67 (2011) 4807e4813

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Tetrahedron
journal homepage: www.elsevier.com/locate/tet

Six new monoterpenoid indole alkaloids from the aerial part of Gelsemium elegans
Sheng Ouyang a, d, y, Lei Wang b, y, Qing-Wen Zhang c, Guo-Cai Wang b, Ying Wang b, Xiao-Jun Huang b,
Xiao-Qi Zhang b, Ren-Wang Jiang b, Xin-Sheng Yao b, Chun-Tao Che e, Wen-Cai Ye a, b, *
a
Department of Phytochemistry, China Pharmaceutical University, Nanjing 210009, China
b
Institute of Traditional Chinese Medicine and Natural Products, Guangdong Province Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs Research,
Jinan University, Guangzhou 510632, China
c
Institute of Chinese Medical Sciences, University of Macau, Taipa, Macau, China
d
Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China
e
Department of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, University of Illinois at Chicago, Chicago, IL 60612, USA

a r t i c l e i n f o a b s t r a c t

Article history: Six new monoterpenoid indole alkaloids along with four known analogues were isolated from the aerial
Received 22 February 2011 part of Gelsemium elegans. Their structures with absolute configurations were elucidated by NMR,
Received in revised form 24 April 2011 HRESIMS, X-ray diffraction, CD spectra, and molecular modeling calculation. Among them, gelselenidine
Accepted 9 May 2011
(1) is a new gelsedine-type alkaloid with a 2,3-epoxybutane unit. Gelseziridine (2) is the first example of
Available online 13 May 2011
monoterpenoid indole alkaloids with an oxaziridine residue. Compounds 6 and 7 are a pair of N4 epimers
of humantenine N4-oxide. A plausible biogenetic pathway for compounds 1e4 was also proposed.
Keywords:
Ó 2011 Elsevier Ltd. All rights reserved.
Monoterpenoid alkaloid
Oxindole alkaloid
Gelsemium elagans
Gelselenidine
Gelseziridine

1. Introduction 2. Results and discussion

Gelsemium elegans (Loganiaceae), widely distributed in South- 2.1. Structural elucidation of alkaloids 1e7
east Asia, is a well known toxic plant. The roots of this plant have
been used as a traditional Chinese medicine in the treatment of Compound 1 was found to have the molecular formula
cancer, furuncle, and carbuncle.1 Previous phytochemical studies of C21H24N2O4 from its HRESIMS data (m/z 369.1816 [MþH]þ). The 1H
this plant had led to the isolation of more than 70 alkaloids,2,3 NMR spectrum displayed some readily assignable signals due to the
which demonstrated cytotoxic,4 analgesic, and anti-inflammatory5 known gelsenicine (8) portion,6 such as four aromatic protons [dH
activities. These alkaloids attracted much attention of organic 7.53 (1H, d, J¼7.3 Hz, H-9), 7.26 (1H, dd, J¼7.6, 7.6 Hz, H-11), 7.07
chemists and pharmacologists due to their complex structural fea- (1H, dd, J¼7.6, 7.3 Hz, H-10), 6.88 (1H, d, J¼7.7 Hz, H-12)], an
tures and multiple biological effects. In our studies on the bioactive N1-methoxy group [dH 3.93 (3H, s)], and three oxygenated protons
alkaloids from the aerial parts of G. elegans growing in Guangdong [dH 4.27 (2H, m, H2-17), 3.72 (1H, d, J¼4.6 Hz, H-3)]. In addition,
province of China, six new monoterpenoid indole alkaloids gelse- signals for a methyl [dH 1.49 (3H, d, J¼5.4 Hz)] and an oxygenated
lenidine (1), gelseziridine (2), gelsemolenines A and B (3 and 4), methine [dH 3.40 (1H, q, J¼5.4 Hz)] were observed. The 13C NMR
(4R)-gelsevirine N4-oxide (5), and (4R)-humantenine N4-oxide (6), spectrum of 1 demonstrated signals similar to those of gelsenicine
together with four known alkaloids (7e10) were isolated (Fig. 1). In (8),6 except for the changes at C-18 and C-19, as well as two addi-
this paper, we describe the structure elucidation of these new al- tional signals at dC 14.1 and 58.8 were observed. The assignment of
kaloids by means of NMR, HRESIMS, X-ray diffraction, circular di- the additional signals could be deduced by 1H, 1H COSY, and HMBC
chroism spectroscopy, and molecular modeling calculation. experiments (Fig. 2). Hence, the 1H, 1H correlation between H3-24
(dH 1.49) and H-23 (dH 3.40), as well as the HMBC cross peaks be-
tween H3-24 (dH 1.49) and C-19 (dC 59.1), between H-23 (dH 3.40)
* Corresponding author. Tel./fax: þ86 20 8522 1559; e-mail address: chywc@
and C-20 (dC 182.2), between H3-18 (dH 1.70) and C-20, as well as
yahoo.com.cn (W.-C. Ye). between H3-18 and C-23 (dC 58.8) revealed the presence of a four-
y
These authors contributed equally to this work. carbon side chain (Fig. 2). According to the chemical shifts of C-19,

0040-4020/$ e see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.tet.2011.05.026
4808 S. Ouyang et al. / Tetrahedron 67 (2011) 4807e4813

Fig. 1. Structures of alkaloids 1e7.

C-23, and H-23, as well as the degree of unsaturation and the On the other hand, the distances calculated from H-23 to H-14a and
characteristic IR absorptions at 1248 and 885 cm1 for epoxy,7 a C- H-14b of the 19S,23S isomer were too large (>4.0  A) to produce
19, C-23 epoxy group was assigned. NOE correlations. Thus, the configuration (19R,23R) of 19, 23-epoxy
group was determined. The CD spectrum of 1 showed the same
Cotton effects as the known gelsedine-type alkaloids, such as gel-
senicine (8) and 19-oxo-gelsenicine,9 indicating the presence of S
configuration at C-7 (Fig. 4). Therefore, the structure including the
absolute configuration of 1 was established as shown in Fig. 1 and
named as gelselenidine. It is noteworthy that gelselenidine (1) is
the first example of gelsedine-type alkaloid bearing a 2,3-
epoxybutane residue, which represents a rare class of Gelsemium
alkaloids with an additional C2 unit.10

Fig. 2. Key 1He1H COSY and HMBC correlations of 1.

The configuration of the epoxide at C-19 and C-23 was deduced

by ROESY experiment and molecular mechanics calculation. The
NOE correlation between H3-18 and H3-24 indicated the presence
of a trans-epoxide. Furthermore, the molecular modelings of
19R,23R and 19S,23S isomers were, respectively, submitted to
a conformational analysis using the Tripos force field based on
molecular modeling software package SYBYL 7.0.8 The corre-
sponding minimum geometries were further fully optimized by
using DFT at the B3LYP/6-31G(d) level as implemented in the
Gaussian 09 program package (Fig. 3). The optimized structure of
the 19R,23R isomer was fully consistent with the corresponding
ROESY data. The calculated distances from H-23 to H-14a, H-14b,
and H-15 were 2.29, 2.96, and 2.45 
A, consistent with the ROESY
correlations between H-23 and H-14a, H-14b and H-15 of 1 (Fig. 3). Fig. 4. CD spectra for 1e8 (in MeOH).

Fig. 3. Two DFT-optimized structures (19R,23R- and 19S,23S-isomers) and key ROESY correlations for 1.
 S. Ouyang et al. / Tetrahedron 67 (2011) 4807e4813 4809

The molecular formula of 2 was established as C19H22N2O4 by Table 2

13
a quasi-molecular ion at m/z 343.1659 [MþH]þ (calcd m/z 343.1652) C (100 MHz) NMR data for compounds 1e7 in CDCl3

in its HRESIMS. The IR spectrum showed absorptions due to car- Position 1 2 3 4 5 6 7

bonyl (1729 cm1) and aromatic ring (1616, 1466 cm1). The UV 2 171.3 171.5 171.7 171.5 171.8 173.6 173.6
spectrum of 2 displayed maxima absorptions at 208 and 250 nm, 3 74.7 74.6 72.0 71.9 68.8 72.5 72.0
characteristic for an indolin-2-one chromophore. The 1H NMR 5 73.1 68.5 47.1 45.9 86.0 77.6 77.1
6 37.7 32.2 35.4 35.4 49.5 27.6 30.3
spectrum of 2 showed signals for an ortho-disubstituted benzene
7 55.9 55.1 53.0 53.0 51.4 54.9 55.7
ring [dH 7.47 (1H, d, J¼7.4 Hz), 7.28 (1H, dd, J¼7.7, 7.7 Hz), 7.09 (1H, 8 132.0 131.9 126.8 126.5 126.4 130.3 127.8
dd, J¼7.7, 7.4 Hz), 6.93 (1H, d, J¼7.7 Hz)], a methoxyl [dH 3.99 (3H, 9 124.6 124.8 126.4 126.4 128.0 125.5 125.7
s)], and an isolated ethyl group [dH 1.06 (3H, t, J¼7.5 Hz), 2.50 (1H, 10 123.4 123.5 123.7 123.7 123.1 123.4 123.5
overlapped), 2.11 (1H, dq, J¼14.8, 7.5 Hz)]. The 13C NMR and DEPT 11 128.1 128.3 128.7 128.8 128.8 128.4 128.7
12 106.6 106.9 107.5 107.6 107.4 107.4 107.6
spectra of 2 displayed 19 signals, including two methyls, four 13 138.0 138.1 139.0 139.1 139.6 138.4 138.7
methylenes, eight methines, one carbonyl, and four quaternary 14 28.1 23.8 137.5 137.6 22.5 26.1 27.1
carbons, suggesting the presence of a monoterpenoid indole alka- 15 37.7 34.1 139.7 139.4 35.2 34.3 33.0
loid. Comparison of the NMR data of 2 with those of gelsedine6 16 39.9 33.6 38.1 38.1 37.5 34.2 30.7
17 62.0 62.0 67.5 67.4 60.5 64.6 65.6
revealed that their chemical shifts were similar, except for the ab-
18 14.2 8.4 8.2 8.2 115.3 13.0 12.9
sence of signals for H-20 and the presence of an oxygenated qua- 19 59.1 22.5 30.6 30.6 134.8 126.0 125.9
ternary carbon signal at dC 93.4 ppm, indicating an oxygen atom 20 182.2 93.4 201.2 201.2 52.2 133.0 131.5
was attached to C-20. The downfield shift (þ8.8 ppm) for C-5 in 2 21/23 58.8 d 169.1 160.0 81.3 60.0 59.7
relative to that in gelsedine6 and the presence of 10 of unsatura- 22/24 14.1 d 23.3 d d d d
N1eOMe 63.3 63.4 63.5 63.6 63.5 63.4 63.5
tion suggested that the oxygen atom was also connected to N4, N4eMe d d d d 53.4 56.5 57.1
forming an oxaziridine ring.11 This conclusion was further con-
firmed by the IR absorption band for oxaziridine ring at
1318 cm112,13 and by the high intensity fragment ion peak at m/z gelsedine. The CD spectrum of 2 showed negative and positive
327.1709 in HRESIMS, which was formed by eliminating a neutral Cotton effects at 209 and 232 nm, respectively (Fig. 4), revealed the
oxygen atom (calcd 327.1703).13 A comprehensive analysis of the presence of S configuration at C-7. Therefore, the structure of 2 was
1
He1H COSY, HSQC, and HMBC spectra allowed the assignment of determined as shown in Fig. 1 and named as gelseziridine. To the
NMR data of 2 as shown in Tables 1 and 2. best of our knowledge, gelseziridine (2) is the first example of
The relative configuration of 2 could be deduced from the ROESY monoterpenoid indole alkaloids bearing an oxaziridine unit. From
experiment and molecular modeling calculations (Gaussian 09). In a biogenetic point of view, gelseziridine (2) may be a biogenetic
the ROESY spectrum, the correlations of H-3/H2-14, H-5/H-16, H-5/ precursor of 18,19-nor-gelsedine-type alkaloids (Scheme 1).10,15
H2-17, H-15/H2-17, and H-15/H3-18 indicated that the relative The HRESIMS of 3 showed a quasi-molecular ion at m/z 385.1763
configuration of C-3, C-5, C-15, and C-16 in 2 is identical to that in [MþH]þ, consistent with the molecular formula C21H24N2O5 (calcd
gelsedine (Fig. 5).6 The a-configuration of the epoxy located at 385.1758), indicating that 3 has two carbon atoms more than the
C20eN4 was indicated by the g-gauche effect from the epoxy group common gelsedine-type alkaloids. The 1H NMR spectrum of 3
to C-16 and C-18 (Fig. 5),14 which resulted in significant upfield showed signals for four aromatic protons [dH 7.47 (1H, d, J¼7.4 Hz),
shifts of C-16 (8.1 ppm) and C-18 (3.6 ppm) compared with 7.35 (1H, ddd, J¼7.8, 7.6, 1.1 Hz), 7.15 (1H, ddd, J¼7.6, 7.4, 1.0 Hz),

Table 1
1
H (400 MHz) NMR data for compounds 1e7 in CDCl3a

Position 1 2 3 4 5 6 7
3 3.72 (d, 4.6) 3.68 (d, 3.9) 4.28 (d, 6.5) 4.29 (d, 6.2) 3.81 (s) 3.58 (d, 7.5) 3.62 (d, 7.1)
5 4.49 (m) 3.96 (m) 4.41 (m) 4.51 (m) 4.01 (br s) 4.18 3.80 (m)
6 a: 2.33 (d, 15.3); 2.29 (2H, m) a: 2.04 (dd, 14.0, 5.2); a: 2.05 (dd, 14.0, 5.3); 3.53 (s) a: 2.37; a: 2.11 (m);
b: 2.43 b: 1.61 (dd, 14.0, 10.1) b: 1.44 (dd, 14.0, 10.1) b: 3.44 b: 2.44
(dd, 15.5, 4.9) (dd, 17.6, 4.8) (dd, 15.3, 9.0)
9 7.53 (d, 7.3) 7.47 (d, 7.4) 7.47 (d, 7.4) 7.46 (d, 7.5) 7.37 (d, 7.6) 7.34 (d, 7.6) 7.34
10 7.07 (dd, 7.6, 7.3) 7.09 (dd, 7.7, 7.4) 7.15 (ddd, 7.6, 7.4, 1.0) 7.15 (ddd, 7.6, 7.6, 1.0) 7.04 (dd, 7.6, 7.6) 7.10 (ddd, 7.6, 7.6, 1.0) 7.11 (dd, 7.6, 7.6)
11 7.26 (dd, 7.6, 7.6) 7.28 (dd, 7.7, 7.7) 7.35 (ddd, 7.8, 7.6, 1.1) 7.37 (ddd, 7.8, 7.6, 1.0) 7.27 (dd, 7.7, 7.7) 7.30 (ddd, 7.7, 7.7, 1.0) 7.32
12 6.88 (d, 7.7) 6.93 (d, 7.7) 7.00 (d, 7.8) 7.01 (d, 7.8) 6.93 (d, 7.7) 6.98 (d, 7.7) 6.99 (d, 7.7)
14 a: 2.29 (d, 15.1); a: 2.43 (d, 15.4); 7.30 (dd, 6.1, 1.8) 7.32 (dd, 6.1, 1.7) a: 2.90 (dd, 14.7, 2.4); a: 2.22 (ddd, 15.4, a: 2.19 (m);
b: 2.11 (ddd, 14.9, b: 2.18 (m) b: 2.05 (ddd, 14.7, 5.6, 2.8) 11.7, 7.6); b: 2.27 (m)
9.5, 4.8) b: 2.37
15 2.92 (t, 9.1) 2.77 (dd, 10.4, 7.6) d d 2.43 (m) 2.67 (m) 2.71 (m)
16 2.56 (t, 8.1) 2.50 3.31 (br s) 3.31 (br s) 2.54 (d, 8.0) 2.49 (m) 3.71 (m)
17 4.27 (2H, m) 4.22 (2H, m) a: 4.26 (d, 8.9); a: 4.27 (dd, 9.8, 1.0); a: 3.99 (br d, 11.3); a: 4.00; 4.14 (2H, m)
b: 3.71 (dd, 9.3, 3.1) b: 3.72 (dd, 9.8, 3.1) b: 4.10 (br d, 11.4) b: 4.18
18 1.70 (s) 1.06 (t, 7.5) 1.21 (t, 7.3) 1.21(t, 7.3) a: 5.22 (d, 11.0); 1.73 (d, 6.9) 1.68 (d, 6.9)
b: 5.02 (d, 17.8)
19 d a: 2.50; a: 3.02 (dq, 17.5, 7.3); a: 3.03 (dq, 17.5, 7.2); 6.18 (dd, 17.8, 11.0) 5.70 (q, 6.9) 5.68 (q, 6.9)
b: 2.11 b: 2.77 (dq, 17.5, 7.3) b: 2.78 (dq, 17.5, 7.2)
(dq, 14.8, 7.5)
21/23 3.40 (q, 5.4) d d 8.07 (s) a: 3.23 (d, 12.1); a: 4.00; a: 4.02 (d, 14.3);
b: 3.55 (d, 12.4) b: 4.78 b: 4.42 (d, 14.3)
(d, 14.6)
22/24 1.49 (d, 5.4) d 1.94 (s) d d d d
N1eOMe 3.93 (s) 3.99 (s) 3.99 (s) 3.99 (s) 3.96 (s) 3.98 (s) 3.95 (s)
N4eMe d d d d 3.31 (s) 3.31 (s) 3.19 (s)
NH d d 6.46 (d, 7.8) 6.55 (d, 7.7) d d d
a
Overlapped signals were reported without designating multiplicity.
4810 S. Ouyang et al. / Tetrahedron 67 (2011) 4807e4813

characteristic for an indolin-2-one chromophore. The 1H NMR

spectrum of 5 showed signals for an ortho-disubstituted benzene
ring [dH 7.37 (1H, d, J¼7.6 Hz), 7.27 (1H, dd, J¼7.7, 7.7 Hz), 7.04 (1H,
dd, J¼7.6, 7.6 Hz), 6.93 (1H, d, J¼7.7 Hz)], a terminal double bond [dH
6.18 (1H, dd, J¼17.8, 11.0 Hz), 5.22 (1H, d, J¼11.0 Hz), 5.02 (1H, d,
J¼17.8 Hz)], and a N1-methoxy group [dH 3.96 (3H, s)]. The 13C NMR
spectrum of 5 displayed twenty-one signals. The signals for a car-
bonyl at dC 171.8 and two quaternary carbons at dC 51.4 and 52.2
were characteristic for gelsemine type alkaloid. Comparison of the
NMR data of 5 with those of the known compound gelsevirine
(9)9,16 revealed that their NMR signals were similar, except for the
resonances for N4eCH3 (dH 3.31, dC 53.4), CH-5 (dH 4.01, dC 86.0), and
CH2-21 (dH 3.55, 3.23, dC 81.3) were downfield shifted in 5. Taking
into account that 5 had an oxygen atom more than the known
compound gelsevirine (9), 5 was proposed to be a N4-oxide de-
rivative of gelsevirine (9). With the aid of 1He1H COSY, HSQC,
HMBC, and ROESY experiments, the 1H and 13C NMR data of 5 were
assigned (Tables 1 and 2). Furthermore, the structure with relative
configuration of 5 was unequivocally confirmed by an X-ray dif-
fraction analysis (Fig. 6). The CD spectrum of 5 displayed negative,
Fig. 5. Key ROESY correlations and the g-gauche effect of epoxy group of 2.
negative, and positive cotton effects at 212, 258, and 235 nm, re-
spectively, indicating the absolute configuration of C-7 was also S
7.00 (1H, d, J¼7.8 Hz)], an oxygenated methylene [dH 4.26 (1H, d, (Fig. 4).3 Based on the above evidences, the structure of 5 was de-
J¼8.9 Hz), 3.71 (1H, dd, J¼9.3, 3.1 Hz)], an oxygenated methine [dH termined as (4R)-gelsevirine N4-oxide.
4.28 (1H, d, J¼6.5 Hz)], a N1-methoxyl [dH 3.99 (3H, s)], and an ethyl Compound 6 displayed the molecular formula C21H26N2O4 by its
[dH 1.21 (3H, t, J¼7.3 Hz), 3.02 (1H, dq, J¼17.5, 7.3 Hz), 2.77 (1H, dq, HRESIMS (m/z 371.1966 [MþH]þ). The 1H and 13C NMR data of 6
J¼17.5, 7.3 Hz)], suggesting the presence of a monoterpenoid indole were similar to those of humantenine,17 except for the chemical
alkaloid. The NMR data at dC 137.5 (C-14), 139.7 (C-15), 201.2 (C-20) shifts around the N4 atom, which suggested that 6 is an N4-oxide
and dH 7.30 (H-14) are characteristic for an a,b-unsaturated ketone derivative of humantenine. This conclusion was confirmed by the
unit. In the HMBC spectrum, correlations between H-16 (dH 3.31) extensive analysis of 1He1H COSY, HSQC, HMBC, and ROESY spectra.
and C-14 (dC 137.5), C-15 (dC 139.7), and C-20 (dC 201.2), between H- The S configuration at C7 was deduced from the CD spectra of 6
14 (dH 7.30) and C-20, as well as between H-18 (dH 1.21) and C-20 (Fig. 4).17 The configuration at the N4 position was determined to be
suggested that 3 was a N4/C-20 seco-oxindole alkaloid,15 which was R by the lower-field-shifted H-6 interpreted from the anisotropy
further confirmed by comparison of the NMR data of 3 with those of effect of the oxygen atom on N4, as well as the ROESY cross peaks
gelseiridone.15 The remaining NMR signals at dC 169.1 and 23.3, as between N4eCH3 (dH 3.31) and H-5 (dH 4.18) and H-16 (dH 2.49)
well as dH 1.94 (3H, s) could be assigned to an acetyl group. The (Fig. 7). Therefore, the structure of 6 was elucidated as (4R)-
linkage between the acetyl group and N4 atom was demonstrated humantenine N4-oxide.
by HMBC cross peak between H-5 (dH 4.41) and C-21 (dC 169.1). Compound 7 showed the same molecular formula as 6 by its
The relative stereochemistry of 3 was deduced by analysis of its HRESIMS C21H26N2O4 (m/z 393.1780 [MþNa]þ). The 1H and 13C
ROESY spectrum.15 The absolute configuration of the spiro-center NMR data of 7 were identical to those of humantenine N4-oxide.18
at C-7 was determined to be S by comparison of its CD spectrum However, the absolute configuration of N4 was not clearly defined
with that of gelseiridone (Fig. 4).15 Based on the above evidences, in the previous work. The CD spectrum of 7 displayed similar
the structure of 3 was fully established and named as gelsemo- Cotton effects as that of 6, indicated the presence of S configura-
lenine A. tion at C-7 in 7 (Fig. 4). In the 1H NMR spectrum, the signals for
The molecular formula of 4 was displayed as C20H22N2O5 by its H-16 was downfield (þ1.40 ppm) compared with that of human-
HRESIMS (m/z 393.1422 [MþNa]þ, calcd 393.1421). The 1H and 13C tenine,17 which was attributable to anisotropy of N/O function.9
NMR data of 4 were very similar to those of 3, except for some dif- Furthermore, the ROESY correlations between N4eMe and H-6a/
ferences occurred for the signals due to the group attached to N4 H-6b were observed. Thus, the absolute configuration of N4 was
position. The absence of signals for acetyl group and the presence of assigned as S. The structure of 7 was determined as (4S)-human-
an aldehyde proton [dH 8.07 (1H, s)] in 4 implied that the acetyl group tenine N4-oxide.
in 3 was replaced by a formyl residue in 4, which was further con- The known compounds gelsenicine (8),6 gelsevirine (9),9,16 and
firmed by the HMBC correlation between H-21 (dH 8.07) and C-5 (dC 14-hydroxygelsenicine (10)9 were also isolated and identified on
45.9). With the aid of 1H, 1H COSY, HSQC, HMBC, and ROESY exper- the basis of their physical and spectroscopic data.
iments, the 1H and 13C NMR data of 4 were assigned as shown in
Tables 1 and 2. The absolute configuration of 4 was identical to that of 2.2. Possible biogenetic pathway for alkaloids 1e4
3, since they showed similar Cotton effects in CD measurement
(Fig. 4). Thus, the structure of 4 was determined and named as gel- On the basis of literatures and our research results, a plausible
semolenine B. To the best of our knowledge, gelsemolenines A (3) biosynthetic pathway for new alkaloids 1e4 was proposed
and B (4) are the first examples of monoterpenoid indole alkaloids (Scheme 1). The alkaloid gelsenicine (8), a major component of
possessing an additional acetyl or formyl unit at N4 position. Gelsemium plants, was originated from strictosidine.9 Addition with
Compound 5 was obtained as colorless crystal. The molecular acetaldehyde10 and subsequent oxidation of the olefinic bond
formula of 5 was established as C21H24N2O4 by the quasi-molecular would afford gelselenidine (1). On the other hand, after oxidation of
ion at m/z 369.1810 [MþH]þ (calcd 369.1809) in its HRESIMS. The IR the imine N4/C-2019 of 8, gelseziridine (2) could be yielded. Com-
spectrum showed absorptions for carbonyl (1721 cm1) and aro- pound 2 might be a biogenetic precursor of 18,19-nor-gelsedine-
matic ring (1615, 1592, and 1463 cm1). The UV spectrum of type alkaloids, such as gelsedilam,15 which is characterized by the
5 exhibited maxima absorptions at 209, 256, and 341 nm, oxaziridine cleavage and b-scission.20,21 14-Hydroxygelsenicine
 S. Ouyang et al. / Tetrahedron 67 (2011) 4807e4813 4811

Scheme 1. Plausible biosynthetic pathway for compounds 1e4.

(10) was also derivatized from strictosidine.9 Hydrolytic cleavage of 2.3. Conclusions
the imine part in 10 and dehydration at C-14 and C-15 positions
would afford a ketoamine intermediate I, which was further acet- Six new monoterpenoid indole alkaloids were isolated from the
ylated or formylated to yield gelsemolenines A (3) or B (4). aerial parts of G. elegans. Among them, gelselenidine (1) is a new
The unusual N4/C-20 seco-oxindole alkaloids possessing an gelsedine-type alkaloid with a 2,3-epoxybutane residue; gelsezir-
N4-iridoid residue, such as gelseiridone15 and gelseganine D18 idine (2) has a rare oxaziridine unit at the skeleton of mono-
had been recently isolated from G. elegans. In the possible terpenoid indole alkaloid; gelsemolenines A (3) and B (4) are the
biogenetic pathway of these novel alkaloids, intermediate I first examples of Gelsemium alkaloids with an additional acetyl or
played an important role15,18 (Scheme 1). However, the exis- formyl unit at N4 position. It is very interesting that a pair of N4
tence of the key intermediate I have not been proved. Gelse- epimers of humantenine N4-oxide were co-existed in the same
molenines A (3) and B (4) are obviously biogenetically related to plant. In addition, gelseziridine (2) was proposed to be a biogenetic
I, which could rationalize the existence of I and further support precursor of 18,19-nor-gelsedine-type alkaloids. The presence of
the proposed biosynthetic pathway for the related alkaloids in gelsemolenines A (3) and B (4) could rationalize the biosynthetic
this plant.15,18 pathway proposed for the related alkaloids in this plant.
4812 S. Ouyang et al. / Tetrahedron 67 (2011) 4807e4813

3.2. Plant material

The aerial parts of G. elegans were collected in Shantou city,

Guangdong province of P.R. China, in May of 2009, and authenti-
cated by Prof. Guang-Xiong Zhou (College of Pharmacy, Jinan Uni-
versity). A specimen (No. 2009052301) was deposited in the
Institute of Traditional Chinese Medicine and Natural Products,
Jinan University, People’s Republic of China.

3.3. Extraction and isolation

The air-dried leaves and stems of G. elegans (4.5 kg) were per-
colated five times with 95% EtOH (50 L) at room temperature for 4
days. The combined EtOH solution was concentrated in vacuo to
afford a residue (650 g), which was dissolved in 2% hydrochloric
acid and then successively extracted with petroleum ether and
EtOAc. The remained acidic layer was basified with aqueous am-
monia to around pH 10 and extracted with CHCl3. The CHCl3 so-
lution was concentrated to afford total alkaloids (60 g). The alkaloid
extract was subjected to silica gel column chromatography eluting
with CHCl3/MeOH (100:1/2:1) to afford five major fractions
(AeE). Fraction A (16 g) was resubjected to silica gel column
Fig. 6. Perspective drawing of X-ray structure of 5. chromatography (cyclohexane/EtOAc¼100:0/0:100) to afford 8
(680 mg). Fraction C (24 g) was subjected to column chromatography
over silica gel (cyclohexane/EtOAc/Et2NH¼100:2:1/100:100:2) to

Fig. 7. Key ROESY correlations and the anisotropy effect of N/O function of 6 and 7.

3. Experimental give five fractions (C1eC5). Fraction C1 was further purified by

preparative HPLC (MeOH/H2O/Et2NH¼60:40:0.01, 8 mL min1) to
3.1. General give 1 (8 mg), 2 (11 mg), and 9 (12 mg). Fraction C2 was purified by
preparative HPLC (MeOH/H2O/Et2NH¼48:52:0.01, 8 mL min1) to
Melting points were measured on an XT-5 micro melting afford 3 (8 mg) and 4 (5 mg). Fractions C3 and C5 were purified by
point apparatus. Optical rotations were measured by a Jasco preparative HPLC to give 10 (10 mg) (MeOH/H2O/Et2NH¼58:42:0.01,
P-1020 digital polarimeter at room temperature. UV spectra were 8 mL min1) and 5 (12 mg) (MeOH/H2O/Et2NH¼48:52:0.01,
recorded on a Jasco-V-550 UVevis spectrophotometer. CD spec- 8 mL min1), respectively. Fraction C4 was subjected to column
tra were obtained on a Jasco J-810 spectropolarimeter at room chromatography with C18 reversed-phase silica gel (MeOH/
temperature. IR spectra were conducted on a Jasco FT/IR-480 plus H2O¼30:70/100:0). The 50% MeOH eluent was further purified by
Fourier transform infrared spectrometer. HRESIMS were per- preparative HPLC (MeOH/H2O/Et2NH¼38:62:0.01, 8 mL min1) to
formed on an Agilent 6210 ESI/TOF mass spectrometer. 1D NMR give 6 (9 mg) and 7 (8 mg).
(1H, 13C, and DEPT) and 2D (1He1H COSY, HSQC, HMBC, and
ROESY) NMR spectra were recorded on a Bruker AV-400 spec- 3.4. Characteristics of alkaloids 1e7
trometer with TMS as internal standard, and chemical shifts were
expressed in d values (ppm). Silica gel (200e300 mesh) (Qingdao, 3.4.1. Gelselenidine (1). Colorless gum; [a]D 25
22.9 (c 0.380,
P.R. China), Sephadex LH-20 (Pharmacia Biotec AB), and RP-18 MeOH); CD (c 0.815 mmol/L, MeOH, 22 C) D3 (l, nm): 0 (297),


(Merck, Darmstadt, Germany) were used for column chroma- 0.91 (258), 0 (246), 1.00 (235), 0 (224), 3.07 (211); UV (MeOH):
tographies (CC). Preparative high-performance liquid chroma- lmax nm (log 3)¼208 (7.45), 255 (6.79); IR (KBr): vmax 3417, 2924,
tography (HPLC) was carried on a Varian chromatograph 2852, 1723, 1617, 1466, 1382, 1318,1248, 1113, 1041, 980, 885, 826,
equipped with a Prostar 215 pump and a Prostar 325 UVevis 750 cm1; 1H NMR: see Table 1; 13C NMR: see Table 2; HRESIMS:
detector with a C18 reversed-phase column (Cosmosil, 369.1816 [MþH]þ (calcd for C21H24N2O4, 369.1809).
30250 mm, 5 mm). All solvents used in column chromatography
were of analytical grade (Tianjin Damao Chemical Plant, Tianjin, 3.4.2. Gelseziridine (2). Amorphous powder; [a]D
25
24.5 (c 0.363,
P.R. China). MeOH); CD (c 0.848 mmol/L, MeOH, 22 C) D3 (l, nm): 0 (298),

 S. Ouyang et al. / Tetrahedron 67 (2011) 4807e4813 4813

1.22 (259), 0 (248), 2.01 (232), 0 (221), 4.32 (209); UV (MeOH): 0.0461, RW 0.1248. CCDC-809159 contains the supplementary
lmax nm (log 3)¼208 (7.55), 250 (6.90); IR (KBr): vmax 3421, 2923, crystallographic data for this paper. These data can be obtained free
2853, 1729, 1616, 1466,1318, 1222, 1118, 1040, 953, 882, 750 cm1; of charge from The Cambridge Crystallographic Data Centre via
1
H NMR: see Table 1; 13C NMR: see Table 2; HRESIMS: 343.1659 http://www.ccdc.cam.ac.uk/data_request/cif.
[MþH]þ (calcd for C19H22N2O4, 343.1652).
Acknowledgements
3.4.3. Gelsemolenine A (3). Amorphous powder; [a]D 25
19.4
(c 0.310, MeOH); CD (c 0.807 mmol/L, MeOH, 22 C) D3 (l, nm):

This work was supported by Program for Changjiang Scholars
0 (378), 0.86 (325), 0.30 (294), 6.83 (250), 5.25 (239), 5.99 and Innovative Research Team in University (IRT0965), National
(227), 0 (213); UV (MeOH): lmax nm (log 3)¼206 (7.24), 251 (6.71), Natural Science Foundation of China (Nos. U0932004 and
341 (6.14); IR (KBr): vmax 3441, 3249, 3076, 2971, 2921, 2874, 1726, 81001375), Natural Science Foundation of Guangdong Province
1677, 1635, 1563, 1460, 1191, 1113, 763 cm1; 1H NMR: see Table 1; (Nos. 10451063201005465 and 8351063201000003), and China
13
C NMR: see Table 2; HRESIMS: 385.1763 [MþH]þ (calcd for Postdoctoral Science Foundation (No. 20100470943).
C21H24N2O5, 385.1758).
Supplementary data
3.4.4. Gelsemolenine B (4). Amorphous powder; [a]D 25
10.0
(c 0.251, MeOH); CD (c 0.676 mmol/L, MeOH, 22  C) D3 (l, nm): Supplementary data associated with this article can be found, in
0 (373), 0.93 (324), 0.39 (298), 6.80 (250), 5.36 (237), 6.21 the online version, at doi:10.1016/j.tet.2011.05.026. These data
(226), 0 (212); UV (MeOH): lmax nm (log 3)¼206 (7.26), 268 (6.23); include MOL files and InChIKeys of the most important compounds
IR (KBr): vmax 3314, 2975, 2941, 2874, 1719, 1671, 1617, 1462, 1192, described in this article.
1077, 749 cm1; 1H NMR: see Table 1; 13C NMR: see Table 2;
HRESIMS: 393.1422 [MþNa]þ (calcd for C20H22N2O5, 393.1421). References and notes

3.4.5. (4R)-Gelsevirine N4-oxide (5). Colorless crystals (MeOH). Mp: 1. Editorial Committee of Chinese Materia Medica. Chinese Materia Medica
(Zhonghua Benchao); Shanghai Science and Technology: Shanghai, 1999; Vol.
125e126  C; [a]D25 22.3 (c 0.269, MeOH); CD (c 0.733 mmol/L, 17; 213e216.
MeOH, 22  C) D3 (l, nm): 0 (309), 1.34 (289), 0 (274), 4.42 (258), 2. Yamada, Y.; Kitajima, M.; Kogure, N.; Wongseripipatana, S.; Takayama, H. Chem.dAsian
0 (247), 5.33 (235), 0 (223), 8.93 (212); UV (MeOH): lmax nm J. 2011, 6,166e173.
3. (a) Kitajima, M.; Kobayashi, H.; Kogure, N.; Takayama, H. Tetrahedron 2010, 66,
(log 3)¼209 (6.79), 256 (6.20), 341(5.23); IR (KBr): vmax 3403, 2925, 5987e5992; (b) Xu, Y. K.; Yang, S. P.; Liao, S. G.; Zhang, H.; Lin, L. P.; Ding, J.; Yue,
1721, 1615, 1592, 1463, 1384, 1080, 754 cm1; 1H NMR: see Table 1; J. M. J. Nat. Prod. 2006, 69, 1347e1350.
13
C NMR: see Table 2; HRESIMS: 369.1810 [MþH]þ (calcd for 4. Kitajima, M.; Nakamura, T.; Kogure, N.; Ogawa, M.; Mitsuno, Y.; Ono, K.; Yano,
S.; Aimi, N.; Takayama, H. J. Nat. Prod. 2006, 69, 715e718.
C21H24N2O4, 369.1809).
5. Rujjanawate, C.; Kanjanapothi, D.; Panthong, A. J. Ethnopharmacol. 2003, 89,
91e95.
3.4.6. (4R)-Humantenine N4-oxide (6). Amorphous powder; [a]D25 6. Takayama, H.; Tominaga, Y.; Kitajima, M.; Aimi, N.; Sakai, S. J. Org. Chem. 1994,
13.2 (c 0.317, MeOH); CD (c 0.865 mmol/L, MeOH, 22  C) D3 (l, 59, 4381e4385.
7. Kuroda, M.; Aoshima, T.; Haraguchi, M.; Young, M. C. M.; Sakagami, H.; Mimaki,
nm): 0 (305), 3.73 (259), 0 (248), 10.06 (228), 0 (214), 1.62 (209); Y. J. Nat. Prod. 2006, 69, 1606e1610.
UV (MeOH): lmax nm (log 3)¼207 (7.97), 255 (7.19), 341(5.71); IR 8. Ciminiello, P.; Catalanotti, B.; Dell’Aversano, C.; Fattorusso, C.; Fattorusso, E.;
(KBr): vmax 3412, 2924, 1719, 1617, 1465, 1384, 1120, 1072, 750 cm1; Forino, M.; Grauso, L.; Leo, A.; Tartaglione, L. Org. Biomol. Chem. 2009, 7,
1 3674e3681.
H NMR: see Table 1; 13C NMR: see Table 2; HRESIMS: 371.1966 9. Ponglux, D.; Wongseripipatana, S.; Subhadhirasakul, S.; Takayama, H.; Yokota,
[MþH]þ (calcd for C21H26N2O4, 371.1965). M.; Ogata, K.; Phisalaphong, C.; Aimi, N.; Sakai, S. Tetrahedron 1988, 44,
5075e5094.
10. Yamada, Y.; Kitajima, M.; Kogure, N.; Takayama, H. Tetrahedron 2008, 64,
3.4.7. (4S)-Humantenine N4-oxide (7). Amorphous powder; [a]D25 7690e7694.
11.5 (c 0.301, MeOH); CD (c 0.811 mmol/L, MeOH, 22  C) D3 11. Davis, F. A.; Theddu, N.; Edupuganti, R. Org. Lett. 2010, 12, 4118e4121.
(l, nm): 0 (305), 2.80 (257), 0 (244), 8.64 (227), 0 (219), 16.53 12. Singh, B. J. Am. Chem. Soc. 1968, 90, 3893e3894.
13. Xu, J.; Chen, L. Rapid Commun. Mass Spectrom. 1999, 13, 2424e2427.
(208); UV (MeOH): lmax nm (log 3)¼207 (7.19), 255 (6.51), 14. (a) Ju, J.; Liu, D.; Lin, G.; Xu, X.; Han, B.; Yang, J.; Tu, G.; Ma, L. J. Nat. Prod. 2002,
341(4.25); IR (KBr): vmax 3407, 2937, 1720, 1618, 1466, 1386, 1330, 65, 42e47; (b) Xu, L.; Wang, F. P. Tetrahedron 2005, 61, 4467e4474.
1209, 1122, 1065, 944, 886, 752 cm1; 1H NMR: see Table 1; 13C 15. Kogure, N.; Ishii, N.; Kitajima, M.; Wongseripipatana, S.; Takayama, H. Org. Lett.
NMR: see Table 2; HRESIMS: 393.1780 [MþNa]þ (calcd for 2006, 8, 3085e3088.
16. Schun, Y.; Cordell, G. A.; Garland, M. J. Nat. Prod. 1986, 49, 483e487.
C21H26N2O4, 393.1785). 17. Lin, L.; Cordell, G. A.; Ni, C.; Clardy, J. J. Nat. Prod. 1989, 52, 588e594.
18. Yin, S.; He, X. F.; Wu, Y.; Yue, J. M. Chem.dAsian J. 2008, 3, 1824e1829.
3.4.8. Crystal data for (4R)-gelsevirine N4-oxide (5). C21H24N2O4, 19. Bourguet, E.; Baneres, J.; Girard, J.; Parello, J.; Vidal, J.; Lusinchi, X.; Declercq,
J. Org. Lett. 2001, 3, 3067e3070.
monoclinic, space group P21, a 9.33100(1), b 21.0564(3), c 20. Usuki, Y.; Peng, X.; Cu € lgeze, B.; Manyem, S.; Aube , J. Arkivoc 2006, iv, 189e199.
10.9924(2) Ǻ, V 2159.75(6) Ǻ3, Z 2, Dcalcd 1.256 g cm1, T 293 K, R 21. Black, D. S. C.; Johnstone, L. M. Angew. Chem. 1981, 93, 703e704.