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Tetrahedron Letters 57 (2016) 58815884

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Tetrahedron Letters
journal homepage: www.elsevier.com/locate/tetlet

Anti-cancer and anti-inflammatory new vakognavine-type alkaloid from


the roots of Aconitum carmichaelii
Ying Liang a, Jian-Lin Wu a, Xun Li b, Ming-Quan Guo b, Elaine Lai-Han Leung a, Hua Zhou a, Liang Liu a,,
Na Li a,
a
State Key Laboratory of Quality Research in Chinese Medicine, Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology,
Avenida Wai Long, Taipa, Macao
b
Key Laboratory of Plant Germplasm Enhancement and Specialty Agriculture, Wuhan Botanical Garden; Sino-Africa Joint Research Center, Chinese Academy of Sciences,
Wuhan 430074, China

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

Article history: Vakognavine-type alkaloids are a rare kind of diterpenoid alkaloid isolated from Delphinium and Aconitum
Received 3 November 2016 species. During our research on the chemical constituents of the roots of Aconitum carmichaelii, a new
Revised 10 November 2016 vakognavine-type alkaloid (1) along with a known vakognavine-type alkaloid, trifoliolasine E (2), and
Accepted 15 November 2016
two known hetisine-type alkaloids (3 and 4) were obtained. The structures were elucidated by a combi-
Available online 16 November 2016
nation of high resolution ESI-MS, 1D and 2D NMR spectra. Compound 1 showed strong inhibitory effects
on HT-29, SGC-7901, and HepG2 cell lines with IC50 values of 0.948, 0.983, and 3.645 lM, respectively,
Keywords:
while trifoliolasine E (2) exhibited moderate activities. Compound 1 was also found to dose-dependently
Vakognavine-type alkaloid
Aconitum carmichaelii
inhibit the activity of cyclooxygenase-2 (COX-2) with an IC50 value of 29.75 lM. To the best of our knowl-
Anti-cancer activity edge, this is the first report on the bioactivity of vakognavine-type diterpenoid alkaloids.
Anti-inflammatory activity 2016 Published by Elsevier Ltd.

Introduction showed moderate inhibition on HT-29 and HepG2 cell lines with
IC50 values of 9.415 and 66.430 lM, respectively. Additionally,
Vakognavine-type alkaloids are a rare kind of diterpenoid alka- compound 1 inhibited the activity of cyclooxygenase-2 (COX-2)
loid, containing the N,19-secohetisine skeleton and a C-4 aldehyde with an IC50 of 29.75 lM. Herein, we described the separation,
group. The first vakognavine-type alkaloid, vakognavine, was iso- structural elucidation, and the bioactivity of these compounds.
lated from the roots of Aconitum palmatum in 1965. Until now, only
21 naturally occurring vakognavine-type alkaloids have been
reported from Aconitum14 and Delphinium510 species. Moreover, Result and discussion
there is no report on the bioactivity of vakognavine-type alkaloids.
Aconitum carmichaelii Debx. is a widely used traditional Chinese The methanol extract of powdered roots of A. carmichaelii was
medicine in East Asia, which possesses anti-inflammatory, prepared by heating at reflux. The extract was then divided into
analgesic, and anti-cancer activities, and contains rich C19- and four parts by liquid-liquid partition, i.e. a hexane extract, ethyl
C20-diterpenoid alkaloids, including one vakognavine-type.4 The acetate extract, n-butanol extract, and water extract. Using normal
presented research on the hexane extract of the roots of A. carmi- phase column chromatography, reversed phase column chro-
chaelii led to the isolation of one new (1) and one known vakog- matography (ODS), and preparative HPLC, four compounds (14)
navine-type (trifoliolasine E, 2) and two hetisine-type alkaloids were obtained from the hexane extract and the structures identi-
(carmichaeline A, 3 and carmichaeline C hydroxide, 4)8,11,12 fied by mass spectra and NMR spectra (Fig. 1).
(Fig. 1). Compound 1 exhibited strong inhibitory effects on HT- Compound 1 was obtained as a colorless amorphous powder,
29, SGC-7901, and HepG2 cell lines with IC50 values of 0.948, and determined as C43H47NO12 by high resolution electrospray ion-
0.983, and 3.645 lM, respectively. Conversely, trifoliolasine E (2) ization mass spectroscopy (ESI-MS) from the quasi-molecular ion
of [M+H]+ at m/z 770.3183 (calc. 770.3171),13 indicating 21 degrees
of unsaturation.
Corresponding authors. The proton signals at dH 7.54 (2H, dd, J = 8.4, 1.2 Hz, H-20 ,60 ),
E-mail addresses: lliu@must.edu.mo (L. Liu), nli@must.edu.mo (N. Li). 7.46 (1H, t, J = 7.2 Hz, H-40 ), 7.27 (2H, t, J = 7.8 Hz, H-30 ,50 ); 7.76
(2H, dd, J = 8.4, 1.2 Hz, H-200 ,600 ), 7.32 (1H, t, J = 7.2 Hz, H-400 ), and

http://dx.doi.org/10.1016/j.tetlet.2016.11.065
0040-4039/ 2016 Published by Elsevier Ltd.
5882 Y. Liang et al. / Tetrahedron Letters 57 (2016) 58815884

H
BzO HO O
H
13 O
AcO 17 AcO HO
16
OAc 12 OAc
20 11
14
BzO 9 15
BzO CH3 CH2COO
2 1 9
H OR H H
N 10 8 N N+
OH
3 7
5
4
H OH H H OH
6
H
19 18
O
OH OH

1 R=COCH(CH 3)2 3 4
2 R=H

Fig. 1. Structures of compounds 14.

7.09 (2H, t, J = 7.8 Hz, H-300 ,500 ) as well as the acetyl signals at dH spectrum, as well as the cross peak between H-7 and H-6 in the
1
2.06 (3H, s) and 2.01 (3H, s) indicated the existence of two benzoyl H-1H COSY spectrum suggested that the hydroxyl group was at
groups and two acetyl groups. The isobutyryl group was deter- C-7. Thus, the planar structure of compound 1 was determined
mined from the HMBC correlations of the two methyl signals at as 1,11-diacetoxy-2,13-dibenzoyloxy-7-hydroxy-15-isobutanoy-
dH 1.27 (6H, d, J = 6.6 Hz) with the methine at dC 34.4 and the car- loxy-N-methyl-N,19-secohetisan-19-al. The relative configurations
bonyl at dC 179.1 (Fig. 2 and Table 1). Among the remaining 21 car- were studied by NOESY experiments (Fig. 3). The cross peaks
bon signals, an aldehyde at dC 195.5 (C-19), a quaternary methyl at between 4-CH3 (H-18) and H-5; H-5 and H-9; and between H-9,
dC 26.2 (C-18), an N-methyl at dC 33.2, an exocyclic methylene at dC H-11 and H-15 indicated their b-orientation. The NOE correlations
116.0 (C-17), a quaternary vinyl carbon at dC 141.7 (C-16), and of H-20 with H-1 and H-14, and that between H-14 and H-7 con-
three quaternary carbons at dC 43.8 (C-4), 53.7 (C-8), 56.6 (C-10) firmed their a-configuration. The C-13 configuration in hetisine-
indicated that compound 1 should be a vakognavine-type alka- type alkaloids can usually be determined by the coupling constant
loid.1,6,7 The HMBC cross peaks of the methine proton at dH 3.06 between H-13 and H-14, where the larger coupling constant
(H-14) with C-8 and C-10 as well as the methine proton at dH (810 Hz) indicates an a-orientation of the substituent, while the
3.88 (H-20) with C-5 and C-8 established the linkage between C- smaller one (4.55 Hz) indicates a b-orientation.14 The proton
14 and C-20 (Fig. 2). Similarly, the connection of 6CHN(CH3)20- resonance at C-14 of compound 1 was observed as a doublet of
CH was determined from the HMBC cross peaks of C-6 with H-20 doublets with a coupling constant with H-13 of 9.6 Hz; therefore,
and the N-CH3 proton. Furthermore, the quaternary methyl group 13-OBz should point to the a-position (Fig. 3). Compound 1 was
and aldehyde were located at C-4 from their HMBC correlations determined as ()-1b,11a-diacetoxy-2a,13a-dibenzoyloxy-7b-
with C-4 and C-5. Thus, compound 1 was deduced as a vakog- hydroxy-15a-isobutanoyloxy-N-methyl-N,19-secohetisan-19-al.
navine-type alkaloid. Two acetoxyl groups and one benzoyloxy Based on the MS and NMR spectra, compound 2 was identified
group were located at C-1, C-11, and C-13 from the HMBC correla- as 1b,11a-diacetoxy-2a,13a-dibenzoyloxy-7b,15a-dihydroxy-N-
tions of the carbonyls with H-1, H-11, and H-13, respectively. methyl-N,19-secohetisan-19-al (trifoliolasine E), which was previ-
Another benzoyloxy group was substituted at C-2 according to ously obtained from Delphinium trifoliolatum.8
the high-frequency shift of H-2 at dH 5.80, which had a 1H-1H COSY Compound 3 had the molecular formula C31H35NO8 as deter-
correlation with H-3. The isobutanoyloxy group was located at C- mined from the high resolution ESI-MS quasi-molecular ion of
15 based on the HMBC correlation between its carbonyl carbon [M+H] + at m/z 550.2433 (calc. 550.2435). Except for the reso-
and H-15 at dH 5.75, which also showed a HMBC cross peak with nances of a benzoyl and two acetyl groups, the NMR signals indi-
the exocyclic methylene (C-17). The correlations of H-9 with cated that 3 had the skeleton of a hetisine-type C20-diterpenoid
the oxygenated carbon at dC 62.7 (C-7) and C-14 in the HMBC alkaloid. However, in comparison to compound 1, a N,O-hemiacetal
group was observed at dC 91.7 (C-19) and dH 4.91 (1H, brs, H-19)
instead of the aldehyde residue. Using a combination of HSQC,
HMBC, 1H-1H COSY, and NOESY (Table S1), compound 3 was
PhCOO
13 determined as 1b,11a-diacetoxy-2a-benzoyloxy-13a,19b-
dihydroxy-hetisan. This compound was first reported by Jiang
CH3COO 17
12
and co-workers,11 but revised as its trifluoroacetate (carmichaeline
20 11 16 O A trifluoroacetate) by Wang and co-workers by comparison of the
CH3COO 14
chemical shift of H-19 with laxipilostine trifluoroacetate,
9 15
PhCOO 2 1 delgramine and 2-acetylseptentriosine.12 In our isolation, diethy-
O
10 8 lamine was used as the modifier in the mobile phase rather than
N H
trifluoroacetic acid, therefore the nitrogen atom in compound 3
3
4
should not exist as the salt form. This is the first time that this
5 7 OH compound has been isolated in this form. Similarly, compound 4
6
H H was identified as the hydroxide salt of 7b,11a,19-trihydroxy-N-
19
18 methyl-13-(2-methylbutyryloxy)-2 a -propionyloxyhetisanium
O
(carmichaeline C hydroxide) due to the preparation being con-
Fig. 2. Key 1H-1H COSY () and HMBC (?) for compound 1. ducted in alkaline conditions.11,12
Y. Liang et al. / Tetrahedron Letters 57 (2016) 58815884 5883

Table 1
1D and 2D NMR data of compound 1 (CDCl3).
1
Position dH (ppm) (J in Hz) dC (ppm) HMBC H-1H COSY NOESY
1 5.86 (1H, d, 4.2) 70.6 H-20 H-20, H-20 ,60
2 5.80 (1H, ddd, 3.6, 2.4, 4.2) 66.8 H-3
3 1.71 (1H, dd, 3.6, 15.8) 29.7 H-18 H-2
2.11 (1H, dd, 2.4, 15.8)
4 43.8 H-3, H-5, H-18, H-19
5 2.42 (1H, s) 55.8 H-3, H-18, H-20 H-9, H-18
6 3.11 (1H, d, 4.8) 60.4 H-20, N-CH3 H-7 H-5, H-18
7 3.71 (1H, d, 4.8) 62.7 H-9 H-6 H-6, H-14
8 53.7 H-14, H-20
9 3.02 (1H, dd, 2.4, 9.6) 47.7 H-5, H-12, H-14 H-11 H-5, H-11, H-15
10 56.6 H-11, H-14
11 5.36 (1H, m) 73.8 H-13 H-9 H-9, H-15
12 2.70 (1H, m) 44.9 H-17 H-13
13 5.38 (1H, m) 72.8 H-11 H-12, H-14 H-14
14 3.06 (1H, dd, 2.4, 9.6) 37.8 H-9, H-12, H-15 H-13 H-7, H-20
15 5.75 (1H, br s) 68.1 H-12, H-17 H-17 H-9, H-11
16 141.7 H-12, H-15, H-17
17 5.12 (1H, d, 1.8) 116.0 H-12, H-15 H-15 H-15
5.34 (1H, d, 1.8)
18 1.08 (3H, s) 26.2 H-5 H-3, H-5, H-6
19 9.20 (1H, s) 195.5 H-3, H-5, H-18 H-6, H-20, H-20 ,60 , N-Me
20 3.88 (1H, s) 63.0 H-5 H-1, H-14, H-20 ,60
N-CH3 2.32 (3H, s) 33.2 H-14
OCOC6H5-2 164.7 H-20 ,60
10 128.4
20 ,60 7.54 (2H, dd, 1.2, 8.4) 129.4 H-30 ,50 , H-40 H-30 ,50 H-19, H-20, N-Me
30 ,50 7.27 (2H, t, 7.8) 128.2 H-20 ,60 , H-40 H-20 ,60
40 7.46 (1H, t, 7.2) 132.9 H-20 ,60 , H-30 ,50 H-30 ,50
OCOC6H5-13 165.8 H-13, H-20 ,60
100 129.2
200 ,600 7.76 (2H, dd, 1.2, 8.4) 129.2 H-300 ,500 , H-400 H-300 ,500 H-1, 11-OAc
300 ,500 7.09 (2H, t, 7.8) 128.3 H-200 ,600 , H-400 H-200 ,600
400 7.32 (1H, t, 7.2) 133.3 H-200 ,600 , H-300 ,500 H-300 ,500
CH3COO-1 169.8 H-1, CH3COO-1
CH3COO-1 2.06 (3H, s) 21.3
CH3COO-11 170.9 H-11, CH3COO-11
CH3COO-11 2.01 (3H, s) 21.5
(CH3)2CHCOO-15 179.1 H-15, (CH3)2CHCOO
(CH3)2CHCOO 2.71 (1H, m) 34.4 (CH3)2CHCOO
(CH3)2CHCOO 1.27 (3H, t, 6.6) 19.2
(CH3)2CHCOO 1.27 (3H, t, 6.6) 18.9

NMR spectra acquired on a 600 MHz Bruker Ascend NMR spectrometer. Residual chloroform signals used as internal chemical shift references.

Table 2
In vitro cytotoxicities of compounds 1 and 2 against human tumor cell lines.
OAc H
H Compound IC50 (lM)
H H
3 H H H 17 HT-29 SGC-7901 HepG2
2
H 1 H AcO 16
5
OH 15
H 1 0.948 0.090 0.983 0.074 3.645 0.127
18 4 10 11
H 3C 9 2 9.415 1.236 66.430 1.266
OBz 6
7
8
OCOC 3H 7 5-FU 13.355 88.248 287.930
H H 12
19 H BzO
O C H
20 14
N 13
H H in the anti-cancer activity. Additionally, compound 1 also showed
Me H dose-dependent inhibition on COX-2 with an IC50 of 29.75 lM,
which was comparable to that of aspirin (29.30 lM), a known
Fig. 3. Key NOESY correlations (M) for compound 1. non-steroidal anti-inflammatory drug. The results indicated that
compound 1 could be a potential anti-cancer and anti-inflamma-
tory compound.
The cytotoxicities of vakognavine-type alkaloids, 1 and 2, were
evaluated against three human tumor cell lines HT-29 (colon can-
cer), SGC-7901 (gastric cancer), and HepG2 (liver cancer) using 5-
Acknowledgments
fluorouracil (5-FU) as a positive control (Table 2). Compound 1
showed strong inhibitory effects on these three tumor cell lines
This work was supported by the Macao Science and Technology
with IC50 values of 14 lM. The inhibitions of compound 2 on Development Fund (086/2013/A3), Natural Science Foundation of
HT-29 and HepG2 were weaker than that of compound 1, which
China (Grant No. 51428303), and the Sino-Africa Joint Research
indicates that the larger alkyl substituent at C-15 was implicated Project (Grant No. SAJC20160233).
5884 Y. Liang et al. / Tetrahedron Letters 57 (2016) 58815884

A. Supplementary data 5. Kulanthaivel P, Holt EM, Olsent JD, Pelletier SW. Phytochemistry. 1990;29:293.
6. Deng Y, Chen D, Song W. Acta Chim Sinica. 1992;50:822.
7. Manners GD, Wong RY, Benson M, Ralphs MH, Pfister JA. Phytochemistry.
Supplementary data (detailed description of the experimental 1996;42:875.
procedures, IR, MS, 1D and 2D NMR spectra for compounds 1 and 8. Zhou XL, Chen DL, Chen QH, Wang FP. J Nat Prod. 2005;68:1076.
9. Lin XY, Chen QH, Wang FP. Helv Chim Acta. 2009;92:745.
3) associated with this article can be found, in the online version,
10. Chen FZ, Chen QH, Liu XY, Wang FP. Helv Chim Acta. 2011;94:853.
at http://dx.doi.org/10.1016/j.tetlet.2016.11.065. 11. Jiang BY, Lin S, Zhu CG, et al. J Nat Prod. 2012;75:1145.
12. Wang FP, Chen DL, Deng HY, Chen QH, Liu XY, Jian XX. Tetrahedron.
2014;70:2582.
References
13. Compound 1: Colorless amorphous powder. a23:5 D = 8.232 (c = 1.0, CHCl3). IR
(KBr): 2919, 1722, 1720, 1638, 1430, 1372, 1277, 1240. 1H (600 MHz) and 13C
1. Pelletier SW, Iyer KN, Wright LH, Newton MG, Singh N. J Am Chem Soc. NMR (150 MHz): see Table 1. HR-ESI-MS (pos.): 770.3183 ([M+H]+,
1971;93:5942. C43H47NO12; calc. 770.3171).
2. Li L, Zhao JF, Wang YB, Zhang HB. Helv Chim Acta. 2004;87:866. 14. Bessonova IA, Saidkhodzhaeva SA. Chem Nat Compd. 2000;36:419.
3. Wang YB, Huang R, Zhang HB, Li L. Helv Chim Acta. 2005;88:1081.
4. Li SH, Zi TY, Wang XQ. Acta Crystallogr Sect E Struct Rep Online. 2008;64:o1394.

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