Synthesis and anticancer activity of a novel series of 9-O-substituted berberine derivatives: A lipophilic substitute role

Article abstract of DOI:10.1016/j.bmcl.2012.10.098

To alter its hydrophobicity, a series of compounds bearing 9-O-alkyl- or 9-O-terpenyl- substituted berberine were synthesized and evaluated for anticancer activity against human cancer HepG2 and HT29 cell lines. We found that the lipophilic substitute of 9-O-alkyl- and 9-O-terpenyl berberine derivatives plays a role in inhibiting the human cancer cell growth and its activity could be maximized with the optimized substitute type and chain length. Most strikingly, nonetheless, of the six compounds prepared, sample 8, a farnesyl 9-O-substituted berberine, showed either comparable or better cytotoxic activity against human cancer HepG2 cell line than that of berberine. Compound 8 had also shown a 104-fold antiproliferation activity in compare with berberine against human hepatoma HepG2 cell lines after 48 incubation hours. Further, in Hoechst 33258 and annexin V-FITC/PI staining analyses it induced apoptosis in HepG2 cells at lower concentration than that of berberine for 24 h. Take all; farnesyl 9-O-substituted berberine could be a potential candidate for new anticancer drug development.

Full text of DOI:10.1016/j.bmcl.2012.10.098

Bioorganic & Medicinal Chemistry Letters 23 (2013) 305–309
Contents lists available at SciVerse ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
Synthesis and anticancer activity of a novel series of 9-O-substituted
berberine derivatives: A lipophilic substitute role
Chih-Yu Lo a, Lin-Chen Hsu b, Min-Shin Chen c, Yi-Jing Lin c, Lih-Geeng Chen c, Cheng-Deng Kuo d,
Jin-Yi Wu c,
⇑
a Department of Food Science, College of Life Sciences, National Chiayi University, Taiwan, ROC
b St. Martin De Porres Hospital Laboratory, Chiayi, Taiwan, ROC
c Department of Microbiology, Immunology and Biopharmaceutics, College of Life Sciences, National Chiayi University, 300 University Road, Chiayi 60004, Taiwan, ROC
d Department of Medical Research and Education, Taipei Veterans General Hospital, Taipei, Taiwan, ROC
a r t i c l e i n f o
a b s t r a c t
Article history:
To alter its hydrophobicity, a series of compounds bearing 9-O-alkyl- or 9-O-terpenyl- substituted berber-
ine were synthesized and evaluated for anticancer activity against human cancer HepG2 and HT29 cell
lines. We found that the lipophilic substitute of 9-O-alkyl- and 9-O-terpenyl berberine derivatives plays
a role in inhibiting the human cancer cell growth and its activity could be maximized with the optimized
substitute type and chain length. Most strikingly, nonetheless, of the six compounds prepared, sample 8,
a farnesyl 9-O-substituted berberine, showed either comparable or better cytotoxic activity against
human cancer HepG2 cell line than that of berberine. Compound 8 had also shown a 104-fold antiprolif-
eration activity in compare with berberine against human hepatoma HepG2 cell lines after 48 incubation
hours. Further, in Hoechst 33258 and annexin V-FITC/PI staining analyses it induced apoptosis in HepG2
cells at lower concentration than that of berberine for 24 h. Take all; farnesyl 9-O-substituted berberine
could be a potential candidate for new anticancer drug development.
Received 15 July 2012
Revised 2 October 2012
Accepted 23 October 2012
Available online 1 November 2012
Keywords:
Berberine
9-O-substituted berberine
HepG2
HT-29
Cytotoxicity
Flowcytometric analysis
Ó 2012 Elsevier Ltd. All rights reserved.
Millions lives lost to cancer in the world each year and the num-
ber is increasing. Through, greater than 30% of cancer is considered
preventable1 and some forms of cancer are curable no matter when
they are diagnosed; many others are only curable if they are caught
at an early stage. Thus cancer remains a leading life threaten dis-
ease and serious peril to humanity health.2
Human hepatocellular carcinoma (HCC) and colorectal carci-
noma are most common types of malignancy in sub-Saharan Africa
and Southeast Asia like Taiwan and China. It is also the fifth most
common and lethal cancer worldwide and usually treated by sur-
gically removal followed by chemotherapy.3,4 We have devoted
our research effort in synthesizing and testing high efficient,
broad-spectrum and low toxic anticarcinogen compounds for po-
tential new drugs candidates for HCC treatment.
Berberine is a quaternary ammonium salt from the protoberb-
erine group of isoquinoline alkaloids. It is found in such plants as
Berberis, Berberis aristata, Hydrastis canadensis, Phellodendron amu-
rense, Coptis chinensis, Tinospora cordifolia, and to a smaller extent
in Argemone mexicana and Eschscholzia californica.5 It has a wide
range of biochemical and pharmacological effects.6–8 In China
and India, berberine was widely in use as an anticancer drug to
treat hepatoma,9 breast cancer,10 bladder cancer,11 and colon can-
cer.12 In China, especially, this natural alkaloid has been used in the
traditional medicine.13,14 Furthermore, it was reported that berber-
ine possesses significant cytotoxicity against human cancer cell
lines, HepG2 and HT-29. It caused a significant reduction of the S
phase fraction of HepG2 cells15 and an arrest of gastric cancer cell
in the G2/M phase.16 Berberine and its derivatives had also been
evaluated as inhibitor of topoisomerase I and II and demonstrated
with anticancer activity.17–19 In addition, a recent study reported
some effectiveness of 9-O-substituted berberine derivatives selec-
tively inhibit acetylcholinesterase (AChE) activity.20
However, for its hydrophilic in nature, berberine is absorbed
poorly in intestines and thus showed a low inhibiting effect on
suppressing cancer cell growth.21 To enhance its intestine absorp-
tion and inhibitory function, we modified berberine analogues and
screened its cytotoxicity in vitro on human liver HepG2 and colon
HT29 cancer cell lines. Two types of functional groups, terpenyl
and alkyl derivatives were investigated for the structure–activity
relationship (SAR).
In specific, we synthesized and tested compounds of 9-O-
alkylberberine derivatives with increased alkyl chain length (butyl,
octyl, and dodecyl, samples 3, 4 & 5) and terpenyl (isoprenyl,
⇑
Corresponding author. Tel.: +886 5 2717925; fax: +886 5 2717778.
E-mail address: jywu@mail.ncyu.edu.tw (J.-Y. Wu).
0960-894X/$ - see front matter Ó 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.bmcl.2012.10.098

306
C.-Y. Lo et al. / Bioorg. Med. Chem. Lett. 23 (2013) 305–309
(2, 3.8 g) in 79% yield.25 To a solution of berberrubine (2, 1 mmol)
5
4
O
O
O
O
6
in dry CH3CN (10 mL) was added alkylated or terpenylated bro-
mide (1.2 mmol) and the reaction mixture was refluxed for 4–
8 h, and the progress of the reaction was monitored by TLC. Then,
the reaction mixture was cooled to room temperature and the sol-
vent was removed under reduced pressure. The crude product was
chromatographed on a neutral Al2O3 column and eluted with
CHCl3/MeOH (99/1) solvent to afford the desired berberine deriva-
tives 3–8 in quantitative yields (Scheme 1). The structures of ber-
berine derivatives 3–8 were determined by NMR and LC–MS
experiments.26 Full signal assignment of 1H and 13C was carried
out with NMR techniques including DEPT, COSY, HSQC, and HMBC
analyses. All compounds were purified by flash column chroma-
tography and were thoroughly characterized by 1H and 13C NMR
spectroscopy and LC–MS spectroscopy.
The anticancer activity of the compounds studied herein was
rated in half maximal inhibitory concentration (IC50), which is a
measure of the effectiveness of a compound in inhibiting biological
or biochemical function. The data of synthesized berberine deriva-
tives (3–8), in compare with those of berberine (1) and berberru-
bine (2), against two tumor cells HepG2 and HT-29 are
summarized in Table 1, with cisplatin data listed as a positive con-
trol. First of all, we found that the activities of the 9-O-alkyl- and 9-
O-terpenyl berberine derivatives against human cancer HepG2 and
HT-29 cell lines increased with the chain length of the substitute at
the C-9 position. In other words, the change in lipophilicity of the
Cl
Cl
N+
N+
8
1
OH
9
OMe
OMe
13
12
OMe
10
11
1
2
Figure 1. Chemical structures of berberine (1) and berberrubine (2).
geranyl, and farnesyl, samples 6, 7 & 8) substitutes. The effective-
ness of anticancer activity of these compounds was carried out
by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
(MTT) assay.22 Data on berberine (sample 1) and its more hydro-
philic derivative, berberrubine (sample 2, Fig. 1), were used as
baseline for evaluating the performance of the synthetic com-
pounds (3–8). Berberrubine is an isoquinoline alkaloid isolated
from the plant Berberis vulgaris L.23 and readily isolated from ber-
berine by pyrolysis.24 The effects of some potent compounds on
human hepatoma HepG2 cells apoptosis were further evaluated.
The synthetic route of lipophilic 9-O-substituted berberine
derivatives 3–8 are outlined in Scheme 1. Berberine chloride (1,
5.0 g) was heated at 190 °C under vacuum (20–30 mmHg) for
1–2 h underwent selective demethylation to afford berberrubine
O
O
O
Cl
N+
Cl
N+
(ii)
(i)
O
OH
OMe
OMe
OMe
2
1
3, R =
O
Br
4
5
N+
, R =
, R =
O
OR
6, R =
7, R =
8, R =
OMe
Scheme 1. Reagents and conditions: (i) 20–30 mmHg, 190 °C, 1–2 h; (ii) K2CO3, alkyl or terpenyl bromide, acetonitrile, reflux, 4–8 h.
Table 1
Lipophilicity and IC50 values for the in vitro screening of berberine 1, berberrubine 2, berberine derivatives (3–8) and cisplatin against human cancer cell lines for 24 h and 48 h
b
b
Compound
ClogPa
HepG2 IC50
24 h
(
lM)
HT-29 IC50
24 h
(lM)
48 h
8.32 2.11
48 h
8.45 0.35
Berberine (1)
Berberrubine (2)
3
4
ꢀ0.771
ꢀ0.707
0.816
2.932
5.048
11.22 1.77
>50
>20
>50
>50
36.93 9.60
0.87 0.11
0.08 0.02
0.28 0.04
1.75 0.63
0.14 0.07
0.27 0.07
24.10 0.10
1.81 0.70
0.19 0.01
0.26 0.10
4.67 0.54
0.21 0.10
0.21 0.07
82.12 2.31
0.97 0.21
0.10 0.05
0.10 0.03
3.17 0.49
0.17 0.02
0.08 0.02
36.00 3.10
4.41 0.30
0.48 0.06
0.93 0.07
10.76 4.71
1.21 1.14
1.82 0.26
53.28 2.64
5
6
7
8
0.931
2.961
4.993
ꢀ1.684
Cisplatinc
a
b
c
Calculated log value of partition coefficient by ChemDraw Ultra 8.0.
IC50, compound concentration required to inhibit tumor cell proliferation by 50%. The values are means SD of three experiments conducted in triplicate.
Positive control.

C.-Y. Lo et al. / Bioorg. Med. Chem. Lett. 23 (2013) 305–309
307
9-O-alkyl berberine derivatives and their parent molecules, berber-
ine (1) and berberrubine (2). Most strikingly, yet, we noticed com-
pound 8 showed a significant higher cytotoxicity against both
cancer cell lines, especially HepG2 with an IC50 value of 0.08
lM
against IC50 of 8.32 M for berberine. In addition, we did not find
l
significant cell death in normal cell lines BNL CL.2 (murine embry-
onic liver cells) after compounds 5 and 8 treatment, however a
marked effect on cell death (under 20%) were observed only at
the maximum concentration of these compounds (10 lM) after
48 h treatment (data not shown). Thus, compounds 5 and 8 appear
to be capable of inducing cytotoxic effects on human liver cancer
cells and human colon cancer cells without incurring significant
cytotoxic effect on normal murine embryonic liver cells.
In light of its superior cytotoxicity, we extended our study in
comparing the antiproliferation function between 9-O-farnesylb-
erberine (8) and berberine on HepG2 cell for the 24, 48 and 72 h
periods, at different doses (Fig. 2). The results showed, though
effectiveness of both compounds was dose- and time-dependent,
compound 8 exhibited 104-fold superiority to that of berberine.
To further extend our cytotoxicity study, we examined the
apoptosis effect of 9-O-farnesylberberine (8). Firstly, after 24 h
incubation in presence or absence of berberine or compound 8,
stained with Hoechst 33258, the topical morphological changes
of HepG2 cells were examined by fluorescence microscopy
(Fig. 3). Evidently, while in the control experiment, the nuclei
of the human hepatoma HepG2 cells were round in shape and
stained homogenously, the berberine (1) or 9-O-farnesylberber-
ine (8) treated ones exhibited typical morphological features of
apoptosis such as cell shrinkage, chromatin condensation and
DNA fragmented.27 These results demonstrated that both berber-
ine (1) and compound 8 inhibited HCC cell proliferation in vitro.
Furthermore to gain the insight of the apoptosis action of ber-
berine (1) and compound 8, HepG2 cells were first either treated
for 24 h with vehicle alone as control or with different concentra-
tions (0.25, 0.5 or 1.0 lM, respectively) of berberine (1) or com-
pound 8. The samples were then stained with annexin V-FITC
and propidium iodide (PI),28 and the percentage of apoptotic
HepG2 cells was subjected to flow cytometry analysis. As shown
in Figure 4, at 1.0 lM concentration, the estimated percentage of
apoptotic cells dramatically increased in both compounds at early
and late apoptosis. Nonetheless, while the effect of berberine (1)
leveled off at the concentration of 0.25 lM, apoptotic effect under
compound 8 went up with treatment dosage and reached at much
higher frequency than those of berberine (1). We attribute this out-
come to the aforesaid superior cytotoxicity and antiproliferation
ability of 9-O-farnesylberberine (8).
In conclusion, we have synthesized several alkylated and ter-
penylated berberine derivatives to evaluate their efficacy as pref-
erential cytotoxic agents against human cancer HepG2 and HT-29
cells. First of all, we found a lipophilic substitute of 9-O-berberine
may enhance its permeability into cell membrane, and thus
cytotoxicity against human cancer cell lines. Most of all, we
discovered a novel berberine derivative, farnesylated berberine
(8), which demonstrated the strongest cytotoxic activity, among
8 samples we studied herein, on human liver cancer HepG2
cell lines, but moderated for human colon cancer HT-29 cell lines.
Figure 2. Cytotoxic effects of berberine (1) and compound 8 on human hepatoma
HepG2 cell lines. HepG2 cells were treated with various concentrations of berberine
(1) and compound 8 for (A) 24 h, (B) 48 h and (C) 72 h. After treatment, cell viability
was estimated by MTT assay. Data are presented as means SD of at least three
replicates from three independent experiments. c: control.
It could be
development.
a potential candidate for new anticancer drug
Acknowledgments
protoberberinium salts affects the anticancer activity of these
derivatives. However, longer chains than octyl alkyl homologs
decreased the anticancer activity as it evidenced by the moderate
activities of 9-O-dodecylberberine bromides (5). Secondly, 9-O-ter-
penyl berberine derivatives were in general more active than
Gratitude is intended to Dr. Christopher Liu for his valuable edi-
torial revision. This study is supported by Grants from The National
Science Council NSC96-2113-M- 415-003 of Republic of China and
grants V99E2-001 from Taipei Veterans General Hospital, Taiwan.

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C.-Y. Lo et al. / Bioorg. Med. Chem. Lett. 23 (2013) 305–309
Figure 3. (Top) Cell morphological change of human liver cancer HepG2 cell lines after 24 h exposure (untreated) and with 20
lM berberine (1) and 0.5 or 1.0 lM compound
8. (Bottom) Cells were stained with Hoechst 33258 and examined using a Nikon (Tokyo, Japan) fluorescence microscope. Magnification ꢁ400.
Figure 4. Induction of apoptosis by berberine (1) or compound 8. FACS analysis of annexin V-FITC and PI double-stained cells. HepG2 cells were untreated or treated with
0.25–1.0 M berberine (1) or compound 8 after 24 h. After treatment, cells were double-stained with annexin V-FITC/PI and analyzed by flow cytometry. Cells appearing in
l
the lower right quadrant show positive annexin V-FITC staining, which indicates phosphatidylserine translocation to the cell surface, and no DNA staining by PI, which
demonstrates intact cell membranes, both features of early apoptosis.
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309
26. To a solution of berberrubine 2 (1 mmol) in dry acetonitrile (10 mL) was added
various alkyl or terpenyl bromide (1.2 mmol) and the reaction mixture was
refluxed for 4–8 h. The progress of the reaction was monitored by TLC. Then,
the reaction mixture was cooled to room temperature and the solvent was
removed under reduced pressure. The crude product was chromatographed on
a silica gel column and eluted with ethyl acetate/methanol (2/1) solvent to
afford the desired berberine derivatives 3–8. 9-O-Butylberberine bromide (3):
[M–Br]+. 9-O-Isoprenylberberine bromide (6),.yellowish brown solid, yield:
68%; UV (MeOH) kmax (loge): 432 (3.65), 351 (4.25), 268 (4.29), 230 (4.34), 204
(4.43) nm; 1H NMR (500 MHz, DMSO-d6): d 9.78 (s, 1H, H-8), 8.92 (s, 1H, H-13),
8.18 (d, J = 9.1 Hz, 1H, H-12), 7.99 (d, J = 9.1 Hz, 1H, H-11), 7.79 (s, 1H, H-1),
7.08 (s, 1H, H-4), 6.16 (s, 2H, –OCH2O–), 5.62 (t, J = 6.56 Hz, 1H, H-20), 4.94 (t,
J = 6.24 Hz, 2H, H-6), 4.84 (d, J = 7.2 Hz, H-10), 4.06 (s, 3H, –OCH3), 3.20 (t,
J = 6.2 Hz, 2H, H-5), 1.71 (s, 3H, H-40), 1.66 (s, 3H, H-3a0); 13C NMR (125 MHz,
DMSO-d6): d 150.7, 149.8, 147.7, 145.4, 142.4, 138.8, 137.4, 132.9, 130.7, 126.5,
yellowish brown solid, yield: 86%; UV (MeOH) kmax (loge): 431 (3.77), 350
(4.41), 267 (4.43), 231 (4.45), 205 (4.41) nm; 1H NMR (500 MHz, DMSO-d6): d
9.75 (s, 1H, H-8), 8.94 (s, 1H, H-13), 8.20 (d, J = 9.1 Hz, H-12), 7.99 (d, J = 9.1 Hz,
H-11), 7.80 (s, 1H, H-1), 7.09 (s, 1H, H-4), 6.17 (s, 2H, –OCH2O–), 4.95 (t,
J = 6.3 Hz, 2H, H-6), 4.29 (t, J = 6.8 Hz, 2H, H-10), 4.05 (s, 3H, –OCH3), 3.21 (t,
J = 6.3 Hz, 2H, H-5), 1.86 (p, J = 7.2 Hz, 2H, H-20), 1.52 (sex, J = 7.4 Hz, 2H, H-30),
0.99 (t, J = 7.4 Hz, 3H, H-40); 13C NMR (125 MHz, DMSO-d6): d 150.4, 149.8,
147.7, 145.3, 142.9, 137.5, 133.0, 130.7, 126.7, 123.3, 121.7, 120.5, 120.2, 108.4,
105.4, 102.1, 73.9, 57.0, 55.3, 31.5, 26.3, 18.6, 13.8; LC–MS (ESI+, m/z)
123.4, 122.1, 120.5, 120.2, 119.9, 108.4, 105.4, 102.1, 72.5, 70.2, 63.1, 57.0, 55.3,
26.4, 25.4, 17.9; LC–MS (ESI+, m/z) C24H24NO4
+
:
390.22 [M–Br]+. 9-O-
Geranylberberine bromide (7), bright yellow solid, yield: 76%; UV (MeOH)
kmax (log
e
): 433 (3.82), 352 (4.45), 268 (4.47), 230 (4.50), 205 (4.53) nm; 1H
NMR (500 MHz, DMSO-d6): d 9.81 (s, 1H, H-8), 8.92 (s, 1H, H-13), 8.19 (d,
J = 9.1 Hz, 1H, H-12), 8.00 (d, J = 9.1 Hz, 1H, H-11), 7.80 (s, 1H, H-1), 7.09 (s, 1H,
H-4), 6.18 (s, 2H, –OCH2O–), 5.63 (t, J = 6.9 Hz, 1H, H-20), 4.96 (m, 3H, H-6, 60),
4.89 (t, J = 6.9 Hz, 2H, H-10), 4.07 (s, 3H, –OCH3), 3.20 (t, J = 6.3 Hz, 2H, H-5),
1.97 (m, 4H, H-40, 50), 1.64 (d, J = 0.6 Hz, 1H, H-3a0), 1.58 (s, 3H, H-80), 1.50 (d,
J = 0.6 Hz, 1H, H-7a0); 13C NMR (125 MHz, DMSO-d6): d 150.8, 149.8, 147.7,
145.5, 142.3, 137.4, 132.9, 131.0, 130.6, 126.4, 123.6, 123.4, 122.2, 120.4, 120.2,
119.5, 108.4, 105.4, 102.1, 70.0, 57.0, 55.2, 26.3, 25.9, 25.4, 17.5 16.2; LC–MS
(ESI+, m/z) C29H32NO4+: 458.28 [M–Br]+. 9-O-Farnesylberberine bromide (8),
C
23H24NO4+: 378.33 [M–Br]+. 9-O-Octylberberine bromide (4): bright yellow
solid, yield: 74%; UV (MeOH) kmax (log ): 432 (3.79), 350 (4.42), 268 (4.44), 230
e
(4.47), 204 (4.48) nm; 1H NMR (500 MHz, DMSO-d6): d 9.75 (s, 1H, H-8), 8.95
(s, 1H, H-13), 8.19 (d, J = 9.1 Hz, H-12), 7.99 (d, J = 9.1 Hz, H-11), 7.80 (s, 1H, H-
1), 7.09 (s, 1H, H-4), 6.17 (s, 2H, –OCH2O–), 4.95 (t, J = 6.2 Hz, 2H, H-6), 4.28 (t,
J = 6.8 Hz, 2H, H-10), 4.06 (s, 3H, –OCH3), 3.21 (t, J = 6.2 Hz, 2H, H-5), 1.87 (p, J
=7.2 Hz, 2H, H-20), 1.48 (p, J = 7.3 Hz, 2H, H-30), 1.33 (m, 8H, H-40–H-70), 0.88 (t,
J = 6.8 Hz, 3H, H-80); 13C NMR (125 MHz, DMSO-d6): d 150.4, 149.8, 147.7,
145.3, 142.9, 137.5, 133.0, 126.7, 123.3, 121.7, 120.5, 120.2, 108.4, 105.4, 102.1,
bright yellow solid, yield: 65%; UV (MeOH) kmax (loge): 432 (3.68), 351 (4.27),
268 (4.30), 230 (4.34), 203 (4.48) nm; 1H NMR (500 MHz, DMSO-d6): d 9.78 (s,
1H, H-8), 8.92 (s, 1H, H-13), 8.17 (d, J = 9.2 Hz, 1H, H-12), 8.00 (d, J = 9.2 Hz, 1H,
H-11), 7.78 (s, 1H, H-1), 7.07 (s, 1H, H-4), 6.16 (s, 2H, –OCH2O–), 5.62 (t,
J = 7.2 Hz, 1H, H-20), 4.99 (t, J = 7.0 Hz, 1H, H-60), 4.94 (m, 3H, H-6, 100), 4.88 (d,
J = 7.3 Hz, 1H, H-10), 4.05 (s, 3H, –OCH3), 3.19 (t, J = 6.3 Hz, 2H, H-5), 1.88 (m,
8H, H-40, 50, 80, 90), 1.62 (s, 3H, H-3a0), 1.60 (s, 3H, H-120), 1.51 (s, 3H, H-7a0),
1.47 (s, 3H, H-11a0); 13C NMR (125 MHz, DMSO-d6): d 150.9, 149.9, 147.8,
145.6, 142.6, 142.4, 137.4, 134.8, 133.0, 130.7, 126.5, 124.1, 123.6, 123.5, 122.4,
120.5, 120.3, 119.6, 108.5, 105.5, 102.2, 70.0, 57.0, 55.4, 26.5, 26.2, 25.9, 25.5,
17.6, 16.3, 15.8; LC–MS (ESI+, m/z) C34H40NO4+: 526.30 [M–Br]+.
27. Wang, G. Y.; Lv, Q. H.; Dong, Q.; Xu, R. Z.; Dong, Q. H. J. Asian Nat. Prod. Res.
2009, 11, 219.
74.2, 57.0, 55.3, 31.2, 29.5, 28.8, 28.7, 26.3, 25.3, 22.1, 14.0; LC–MS (ESI+, m/z)
+
C
27H32NO4
:
434.41 [M–Br]+. 9-O-Dodecylberberine bromide (5): dark
yellowish brown solid, yield: 61%; UV (MeOH) kmax (loge): 431 (3.42), 351
(4.03), 267 (4.07), 230 (4.10), 202 (4.19) nm; 1H NMR (500 MHz, DMSO-d6): d
9.77 (s, 1H, H-8), 8.98 (s, 1H, H-13), 8.21 (d, J = 9.1 Hz, H-12), 8.00 (d, J = 9.1 Hz,
H-11), 7.82 (s, 1H, H-1), 7.10 (s, 1H, H-4), 6.18 (s, 2H, –OCH2O–), 4.96 (t,
J = 6.1 Hz, 2H, H-6), 4.28 (t, J = 6.8 Hz, 2H, H-10), 4.05 (s, 3H, –OCH3), 3.21 (t,
J = 6.1 Hz, 2H, H-5), 1.88 (d, J = 7.4 Hz, H-20), 1.47 (q, J = 7.5 Hz, 2H, H-30), 1.29
(m, 16 H, H-40–H-110), 0.86 (t, J = 6.8 Hz, 3H, H-120); 13C NMR (125 MHz,
DMSO-d6): d 150.4, 149.8, 147.7, 145.3, 142.9, 137.5, 133.0, 130.7, 126.7, 123.3,
121.7, 120.5, 120.3, 108.4, 105.5, 102.1, 74.2, 57.0, 55.3, 48.6, 31.3, 29.5, 29.1,
29.1, 28.9, 28.7, 26.3, 25.3, 22.1, 14.0; LC–MS (ESI+, m/z) C31H40NO4+: 490.50
28. Hur, J.-M.; Hyun, M.-S.; Lim, S.-Y.; Lee, W.-Y.; Kim, D. J. Cell Biochem. 2009, 107,
955.