Chinese

Journal of
Natural
Chinese Journal of Natural Medicines 2014, 12(3): 0222−0224 Medicines

A new quinoline alkaloid from the roots of Dictamnus angustifolius

SUN Jian-Bo1, QU Wei1, GUAN Fu-Qin2, LI Lin-Zhen1, LIANG Jing-Yu1*
1
Department of Natural Medicinal Chemistry, China Pharmaceutical University, Nanjing 210009, China;
2
Institute of Botany, Jiangsu Province and Chinese Academy of Science, Nanjing 210014, China
Available online 20 Mar. 2014

[ABSTRACT]
AIM: To investigate the quinoline alkaloids from the roots of Dictamnus angustifolius G.Don ex Sweet (Rutaceae).
METHOD: The quinoline alkaloids were isolated by various column chromatographic methods and their structures were eluci-
dated on the basis of spectral analysis.
RESULTS: A new quinoline alkaloid, 5-methoxylrobustine (1), along with five known quinoline alkaloids were obtained, and
their structures were identified as dictamnine (2), robustine (3), isopteleine (4), γ-fagarine (5), and skimmianine (6). Cytotoxicity
testing of these alkaloids showed that all of them had weak cytotoxic activities against human breast cancer cells (MCF7).
CONCLUSION: Compound 1 is a new quinoline alkaloid. Alkaloid 3 showed stronger anti-proliferation effect than the other al-
kaloids.

[KEY WORDS] Dictamnus angustifolius; Quinoline alkaloids; 5-Methoxylrobustine; Cytotoxicity

[CLC Number] R284.1 [Document code] A [Article ID] 2095-6975(2014)03-0222-03

breast cancer cells. A new compound 5-methoxylrobustine

Introduction
Dictamnus angustifolius G.Don ex Sweet (Rutaceae) is
widely distributed in Xinjiang province of China. The root
bark has been used as an alternative for the traditional medi-
cine, D. dasycarpus, another species from the genus Dictam-
nus L., in the treatment of rheumatism, bleeding, itching,
jaundice, chronic hepatitis, skin diseases, and as an
anti-inflammatory agent, febrifugal, and detoxicant drug [1].
The main chemical constituents include quinoline alkaloids,
limonoids, coumarins, and steroids [2]. As part of ongoing
research for components with structural and biological diver-
sity from the root bark of D. dasycarpus, a comparative phy-
tochemical investigation of D. angustifolius has been per-
formed. Pharmacological studies showed that quinoline alka-
loids have the anti-fungal, cytotoxic, anti-platelet aggregation,
and vasorelaxing effects [3-5]. In order to further explore their
cytotoxic effect, MCF7 cells using the MTT method were
used to research their anti-proliferation activity on human
[Received on] 29-Jan.-2013
[Research funding] This project was supported by the National New
Drug Innovation Major Project of China (No. 2011ZX09307-002-02)
[*Corresponding author] LIANG Jing-Yu: Prof., E-mail: jyliang08
@126.com
These authors have no conflict of interest to declare.
Published by Elsevier B.V. All rights reserved
(1), along with five known quinoline alkaloids were isolated,
in which compound 3 was isolated from this species for the
first time. In this paper, the isolation and structural identifica-
tion of these six quinoline alkaloids and their cytotoxic ef-
fects are described.
Results and Discussion
Alkaloid 1 was obtained as a yellowish powder. The
molecular formula of C13H11NO4 was elucidated based on the
positive HR-TOF-MS pseudo-molecular ion peak at m/z
246.076 2 [M + H]+ (Calcd. 246.076 6). The IR absorption at
3 408 cm−1 showed the presence of a hydroxyl group. The 1H
NMR spectrum (Table 1) was very informative, with a hy-
droxyl signal (δH 6.32) and two ortho-coupled aromatic pro-
tons (δH 7.16 and 7.96). Two singlets at δH 4.21 and δH 4.41
confirmed the presence of methoxyl groups at C-5 and C-4.
Protons at δH 7.03 and δH 7.56 were assigned to H-11 and
H-12, respectively. 13C NMR and DEPT spectra of 1 indicated
thirteen resonances ascribed to two methoxyl groups, six dou-
ble-bond methine carbons, and five double-bond quaternary
carbons. Comparison of the NMR spectroscopic data of 1 and
the known robustine [2] established that 1 had a similar structure
to the latter, but with a methoxyl group replacing 5-H.
To confirm the location of the hydroxyl and two meth-
oxyl groups, HSQC and HMBC experiments were conducted.
The HMBC (Fig. 1) spectrum indicated that the protons at δH

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7.16 (d, J = 9.2 Hz) and δH 7.96 (d, J = 9.2 Hz) were assigned hydroxylfuroquinoline, and named 5-methoxylrobustine. Its
1
to H-7 and H-6, respectively. C-4 (δC 157.5) and the correla- H- and 13C-NMR spectra (Table 1) were completely as-
tions from OCH3 (δH 4.41) to C-4, C-11 suggested that the signed by detailed 2D-NMR experiments.
OCH3 (δH 4.41) was attached to C-4. Meanwhile, C-5 (δC
138.6) and the correlations from the OCH3 (δH 4.21) to C-5
and C-6, from H-6 to C-5 and C-4, and from H-7 to C-8 and
C-9, supported the OCH3 (δH 4.21) attached to C-5. C-8 (δC
149.1) and the neighbor relationship between H-6 and H-7
substantiated the remaining hydroxyl being connected to C-8.
Thus alkaloid 1 was elucidated to be 4,5-dimethoxyl-8- Fig. 1 The HMBC correlations of alkaloid 1
1
Table 1 H (500 MHz) and 13C (125 MHz) NMR spectral data for alkaloid 1 (CDCl3)
Proton δHa, mult., int Carbon δCa Carbon δCa
H-6 7.96, d, 9.2, 1H C-2 164.0 C-9 140.6
H-7 7.16, d, 9.2, 1H C-3 101.7 C-10 114.4
H-11 7.03, d, 2.8, 1H C-4 157.5 C-11 104.7
H-12 7.56, d, 2.8, 1H C-5 138.6 C-12 142.7
8-OH 6.32, s, 1H C-6 118.7 4-OMe 59.0
5-OMe 4.21, s, 3H C-7 114.6 5-OMe 61.9
4-OMe 4.41, s, 3H C-8 149.1
a
Signal assignments are based on 2D-NMR (HSQC, HMBC) spectra

Alkaloids 2-6 were characterized as dictamnine, robustine
, isopteleine [7-9], γ-fagarine [2, 10], and skimmianine [11], re-
[2, 6]
spectively.
The cytotoxicity assay was performed by the modified
MTT method. Briefly, 5.0 × 104/mL cells were seeded into
96-well plates and allowed to adhere for 24 h. Each test
compound was dissolved in DMSO and diluted to 100
μmol·L−1 with complete medium for inhibition rate (IR)
determination. After incubation at 37 ºC in a CO2 atmosphere
for 72 h, MTT solution (5 mg·mL−1) was added, and the plate
was further incubated at 37 ºC for 4 h. The supernatant was
removed before adding DMSO (100 µL) to each well. The
inhibition rate was calculated as IR = (1 − ODtest/ODcontrol) ×
100%. 5 fluorouracil (5-Fu) was used as positive control.
Six quinoline alkaloids showed weak cytotoxic effect.
Alkaloid 3 showed stronger anti-proliferation effect than the
other alkaloids, but far less than 5-Fu (55.6%) under the same
conditions.
Table 2 Inhibitory effects of alkaloids 1−6 on human breast
cancer cells (MCF7)
IR (%)
Cell line
1 2 3 4 5 6
MCF7 18.65 19.67 26.25 14.86 22.81 23.13
Experimental
General experimental procedures
IR spectra were measured on a Nicolet Impact-410 spec-
trometer as KBr pellets, in cm−1. NMR spectra were recorded
on Bruker ACF-300 NMR and ACF-500 NMR spectrometers
with TMS as internal standard. MS spectra were run on
Agilent 1100 Series LC/MSD Trap ESI and Agilent 1100
LC/TOF MSD spectrometers. All solvents used were of ana-
lytical grade (Tianjin Chemical Co., Ltd., Tianjin, China).
Column chromatography was performed on Silica gel H
(100−200 mesh, 200−300 mesh, Qingdao Marine Chemical
Co., Ltd, Qingdao, China) and Sephadex LH-20 (40−75 μm,
Pharmacia, New Jersey, USA). Thin-layer chromatography
was performed on silica gel GF254 (Qingdao Marine Chemical
Co., Ltd., Qingdao, China).
Plant material
The air-dried roots of D. angustifolius were collected
from the Xinjiang Altay Region in 2010 and identified by
Prof. PU She-Ban, China Pharmaceutical University (Nan-
jing, China). A voucher specimen (No. 20100507) is depos-
ited in the Department of Natural Medicinal Chemistry,
China Pharmaceutical University, Nanjing, China.
Extraction and isolation
The air-dried roots of D. angustifolius (18 kg) were ex-
tracted with 80% EtOH under reflux three times. The com-
bined extracts were concentrated and then suspended in water,
followed by partitioning with petroleum ether, CH2Cl2, and
n-BuOH, respectively. The CH2Cl2 fraction (150 g) was frac-
tionated by silica gel column chromatography (100−200
mesh) using an elution gradient (CH2Cl2−CH3OH, 100 : 1 to
1 : 1) to afford ten fractions (I-X). Subfraction VII
(CH2Cl2−CH3OH, 30 : 1, 6.80 g) was subjected to repeated
column chromatography on silica gel and Sephadex LH-20,
and purification by preparative TLC with petroleum
ether-acetone (1 : 1) to yield 1 (27 mg), 2 (45 mg), 3 (11 mg),
4 (23 mg), 5 (17 mg), and 6 (14 mg).

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Cite this articls as: SUN Jian-Bo, QU Wei, GUAN Fu-Qin, LI Lin-Zhen, LIANG Jing-Yu. A new quinoline alkaloid from the
roots of Dictamnus angustifolius [J]. Chinese Journal of Natural Medicines, 2014, 12(3): 222-224

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