Novel carbohydrate modified berberine derivatives: Synthesis and: In vitro anti-diabetic investigation

Article abstract of DOI:10.1039/c9md00036d

Berberine is a bioactive alkaloid used in Chinese medicine and has numerous positive effects on biological systems. This paper describes the facile and highly efficient synthesis of some carbohydrate modified berberine derivatives, and conjugation of the carbohydrate moiety with berberine was finished by "click" chemistry. The cytotoxicity and anti-diabetic measurements of all berberine derivatives were accomplished on HepG2 cell lines, and the results indicated that most of the derivatives exhibit higher anti-diabetic activity than berberine. The mannose modified berberine derivative has significantly lower cytotoxicity than berberine, and the induced IC₅₀ value of this derivative is nearly 1.5 times that of berberine. Furthermore, this mannose modified berberine derivative exhibits high anti-diabetic activity at both high and low drug concentrations, thereby indicating its potential application for the development of novel anti-diabetic drugs.

Full text of DOI:10.1039/c9md00036d

View Article Online
View Journal
MedChemComm
Accepted Manuscript
This article can be cited before page numbers have been issued, to do this please use: L. Han, W. Sheng,
X. Li, S. Attila, H. Lin, K. Liu and L. Wang, Med. Chem. Commun., 2019, DOI: 10.1039/C9MD00036D.
This is an Accepted Manuscript, which has been through the
Volume 7 Number 1 January 2016 Pages 1–204
Royal Society of Chemistry peer review process and has been
accepted for publication.
MedChemComm
Accepted Manuscripts are published online shortly after
acceptance, before technical editing, formatting and proof reading.
Broadening the field of opportunity for medicinal chemists
www.rsc.org/medchemcomm
Using this free service, authors can make their results available
to the community, in citable form, before we publish the edited
article. We will replace this Accepted Manuscript with the edited
and formatted Advance Article as soon as it is available.
You can find more information about Accepted Manuscripts in the
author guidelines.
Please note that technical editing may introduce minor changes
to the text and/or graphics, which may alter content. The journal’s
Themed issue: Antibiotic Resistance
ISSN 2040-2503
standard Terms & Conditions and the ethical guidelines, outlined
in our author and reviewer resource centre, still apply. In no
event shall the Royal Society of Chemistry be held responsible
for any errors or omissions in this Accepted Manuscript or any
consequences arising from the use of any information it contains.
rsc.li/medchemcomm

Please do not adjust margins
Page 1 of 10
MedChemComm
View Article Online
DOI: 10.1039/C9MD00036D
Journal Name
ARTICLE
Novel carbohydrate modified berberine derivatives: Synthesis and
in vitro anti-diabetic investigation
Received 00th January 20xx,
Accepted 00th January 20xx
Liwen Han,a,§ Wenlong Sheng,a,§ Xiaobin Li,a Sik Attila,b Houwen Lin,c Kechun Liu,a,* Lizhen
Wang,a,*
DOI: 10.1039/x0xx00000x
www.rsc.org/
Berberine is a bioactive alkaloid used in Chinese medicine and has numerous positive effects on biological systems. This
paper describes the facile and high efficient synthesis of some carbohydrate modified berberine derivatives, and
conjugation of the carbohydrate moiety with berberine was finished by “click” chemistry. The cytotoxicity and anti-
diabetic measurements of all berberine derivatives were accomplished on HepG2 cell lines, and the results indicated that
most of the derivatives exhibit higher anti-diabetic activity than berberine. The mannose modified berberine derivative has
significantly lower cytotoxicity than berberine, and the induced IC50 value of this derivative is nearly 1.5 folds of
berberine. Furthermore, this mannose modified berberine derivative exhibits high anti-diabetic activity at both high and
low drug concentrations, thereby indicating its potential application for the development of novel anti-diabetic drug.
17
inhibition of PPARγ and C/EBPα,16,
and down regulation the
PEPCK and G6Pase gene expression18 et al.
Introduction
Berberine (Figure 1), an active isoquinoline alkaloid extracted
4
A
1
5
3
2
from Chinese herb Coptischinensis,1 has been widely used for the
O
O
6
Cl
B
3
treatment of gastrointestinal infection and diarrhea in China.2,
7
N
8
Recent studies revealed that berberine has various pharmacological
activities such as anti-microbial,4 anti-diabetic,5 anti-hyperlipidemic
action,6 anti-tumor,7 anti-inflammatory,8 anti-parasitic.9 As an anti-
diabetic agent, berberine obviously reduces the blood glucose in
type 2 diabetic patients and has been optionally used by physicians
in China for several years.10, 11 It has been proved that berberine can
inhibit the activity of α-glucosidase, which hydrolyzes the
oligosaccharides and polysaccharides to release free glucose,
thereby leading to the inefficient absorption rates of glucose, as
shown in Figure 2.12 Furthermore, berberine can activate AMP-
activated protein kinase (AMPK), an important fuel gauge protein
C
O
O
9
13
D
12
10
11
Fig. 1 Chemical structure of berberine
Glucose
absorption
-Glucosidase
AMPK
Glycolysis
Fasting blood
glucose
Mitochondrial
function
Berberine
that monitors the energy levels in mammalian cells, which might
Blood TC, TG
14
partially contribute to the glucose consumption.13,
There are
PPAR and
C/EBP
Adipogenesis
some other proposed mechanisms that can explain the anti-diabetic
activity of berberine, such as mitochondrial function inhibition,15
Liver
Inhibit
PEPCK and
G6Pase
gluconeogenesis
activate
Fig. 2 Proposed anti-diabetic mechanisms of berberine
a. Biology Institute, Qilu University of Technology (Shandong Academy of Sciences),
Jinan 250014, Shandong Province, China.
Therefore, berberine is a promising medicine for treatment of
type 2 Diabetes. But the problem is that oral administration of
berberine usually gives extremely low bioavailability (<5%),10 while
increasing the oral dose and parenteral drug delivery usually cause
severe side effects. Consequently, several novel strategies have
been developed for improving the bioavailability of berberine,
which alternatively enhance its anti-diabetic activity. The most
important strategy is structural modification of berberine to provide
b. Institute of Transdisciplinary Discoveries, University of Pecs, Pécs, Ifjúság útja 20,
7624, Hungary.
c. Research Center for Marine Drugs, State Key Laboratory of Oncogenes and
Related Genes, Department of Pharmacy, Ren Ji Hospital, School of Medicine,
Shanghai Jiao Tong University, Shanghai 200127, China
§These authors contributed equally to this paper.

Corresponding authors. Tel.: +8613869197581; e-mail: wlzh1106@126.com
(Lizhen Wang), liukechun2000@163.com (Kechun Liu).
Electronic Supplementary Information (ESI) available: [details of any
supplementary information available should be included here]. See
DOI: 10.1039/x0xx00000x
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 1
Please do not adjust margins

Please do not adjust margins
MedChemComm
Page 2 of 10
ARTICLE
Journal Name
novel fully-synthetic anti-diabetic drug candidates.19 For example,
the 9-O-hydrophilic and hydrophobic modification of berberine can
both improve its bioavailability and enhance the anti-diabetic
View Article Online
OH
R
R =
OH
HO
DOI: 10.1039/C9MD00036D
N
N
O
O
O
O
HO
HO
Cl
HO
N
HO
1
HO
N
2
20
activity,19,
some C-8,13-substituted berberine derivatives have
O
HO
O
OH
O
significant glucose-lowering property,4, and the 9-OH or 10-OH
pseudoberberines also exhibited high anti-diabetic activity.22 But,
there is no berberine derivative showed potential application as
anti-diabetic drug, and many of these derivatives are unstable
under mild conditions which are easily degraded into berberine and
related analogs in vitro. Consequently, some stable berberine
derivatives should be designed and synthesized to develop effective
anti-diabetic drug candidate.
21
HO
HO
HO
O
HO
OCH3
HO
HO OH
OH
3
4
5
Fig. 4 Carbohydrate modified berberine derivatives 1-5
Results and discussion
Previous research revealed that the presence of carbohydrate
moiety in drugs had the ability to improve their bio-availability and Synthesis of berberine derivatives
receptor-binding affinity, thus resulting in significant drug activity
24
enhancement.23,
Studies on metabolites of berberine in rat
The synthesis routes of glucose and galactose modified berberine
derivatives 1 and 2 were outlined in Scheme 1. The commercially
available berberine chloride was heated at 185-195ºC under high
vacuum (20-30 mm Hg) to produce berberrubine 6.28 Without any
purification, the crude 6 was reacted with propargyl bromide 7 in
DMF to give 8 with an alkynyl group. In parallel, treatment of
pentaacetyl β-D-glucose 9 with HBr (33% in AcOH) afforded glycosyl
bromide 11, which was immediately converted into compound 13
in the presence of NaN3 and DMF with 69% overall yield. Due to the
neighboring group participation effect29-33 of the 2-O-acetyl group
in 11, this reaction was highly stereoselective and gave β-linked
azido sugar 13 as the only product. Thereafter, the acetyl groups in
13 were removed with CH3ONa, and the resultant 15 was subjected
to the Cu–catalyzed “click” reaction with alkylated berberine
derivative 8 to afford the desired compound 1 in 56% total yield.
Using the same protocol, we synthesized the galactose modified
berberine derivative 2 from 10 in four steps with high overall yield
(50%). Therefore, this synthetic routine was highly efficient and
could be applied for the preparation of various carbohydrate
modified berberine derivatives.
revealed that the 9-O-glucosyl-berberine (Figure 3) is existed as the
26
one of the major metabolites.25,
Therefore, synthesis of 9-O-
hydrophilic berberine derivatives to improve its water solubility
20
might be a new idea to increase its bio-availability.19,
This
conjecture was confirmed by Chen and co-workers, who had
synthesized the 9-O-glucosyl-berberine (Figure 3) and proved that
hydrophilic modification of berberine could obviously improve its
bio-availability.19, 20 However, this derivative showed poor stability
under acidic conditions and high temperature (>45 oC).19, 20 In order
to improve the stability of 9-O-glucosyl-berberine and
simultaneously enhance its anti-diabetic activity, we now focused
on the preparation of some carbohydrate modified berberine
derivatives 1–5 (Figure 4). The N-linked glucose, galactose,
mannose, ribose, and rhamnose, which had different content of
hydroxyl groups and water solubility, were chose as the
carbohydrate moiety. The modification was mainly on the C-9
position of berberine, because the key intermediate 9-OH
pseudoberberine (6, Scheme 1) could be easily available through a
pyrolysis strategy. Furthermore, the high efficient “click” chemistry
was used to accomplish the conjugation of monosaccharides with
berberine, which could form a triazole27 linker to give stable
glucosyl berberines. Compared with O-glycosides, the N-glycosides
are stable enough in the presence of glycoside hydrolases under
physiological conditions. The synthesized derivatives were
subjected to the anti-diabetic evaluation in HepG2 cell lines, and
their cytotoxicity activity was determined through Cell Counting Kit-
8 (CCK-8) assay.
Using this high efficient synthetic strategy, we focused on the
synthesis of a mannose modified berberine derivative 3 as shown in
Scheme 2. Our initial work was the synthesis of azido sugar 19α,
through treatment of D-mannose pentacetate 17 with HBr followed
by reaction of the resultant glucosyl bromide 18 with NaN3 in DMF.
While this transformation generated an α,β mixture of 19 with only
29% total yield. As an alternative, we employed a new strategy
based on the Carrington’s method,34, 35 which could directly afford
the azido sugar 19α under mild conditions. Therefore, treatment of
17 with TMSN3 under the promotion of SnCl4 in CH2Cl2 produced
19α with absolute α-selectivity and 88% isolated yield. The high α-
stereoselectivity of this reaction was also caused by the neighboring
participation effect of 2-O-acetyl group in 17. Subsequent
deacetylation of 19α using CH3ONa followed by the Cu–catalyzed
“click” reaction of 20 with 8 under the same condition as described
previously, facilitated the target derivative 3 in 52% overall yield.
This simplified and high efficient strategy was further applied for
the synthesis of rhamnopyranose modified berberine derivative 4
(in three steps and 35% total yield) and ribofuranose modified
berberine derivative 5 (in three steps and 44% total yield).
HO
HO
O
OH
HO
N
O
O
O
OCH3
Fig. 3 9-O-glucosyl-berberine
2 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins

Page 3 of 10
MedChemComm
Please do not adjust margins
Journal Name
ARTICLE
O
O
AcO
AcO
AcO
AcO
AcO
View Article Online
O
O
NaN3
AcO
AcO
AcO
33% HBr in AcOH
DCM
Cl
DOI: 10.1039/C9MD00036D
O
190 o
in a vacuum
C
anhydrous DMF
29%
Cl
O
N
N
OH
OAc
Br
AcO
OCH3
OCH3
17
18
AcO
AcO
OCH3
AcO
AcO
AcO
O
6
O
AcO
+
Berberine chloride
Br
N3
AcO
N3
O
O
19
19
Cl
7
HO
N
CH3ONa
CH3OH
TMSN3, SnCl4
DCM
HO
O
HO
O
19
DMF, 80 o
72% (2 steps)
C
HO
88%
N3
20
OCH3
HO
HO
OH
OH
8
8
OAc
O
R1
NaN3
DMF
OAc
AcO
CuSO4 5H2O
sodium ascorbate
CH3OH
33% HBr in AcOH
DCM
R1
O
R2
AcO
N
O
N
O
R2
OAc
AcO
N
AcO
Cl
Br
N
O
11 R1 = H, R2 = OAc
12 R1 = OAc, R2 = H
52%
(2 steps)
9
R1 = H, R2 = OAc
O
10 R1 = OAc, R2 = H
8
CuSO4 5H2O
sodium ascorbate
CH3OH
OCH3
OAc
R1
CH3ONa
CH3OH
OH
R1
3
O
R2
O
N3
R2
HO
N3
N3
N3
AcO
CH3ONa
TMSN3, SnCl4
AcO
OAc
CH3OH
HO
O
DCM
O
O
13 R1 = H, R2 = OAc
69% (2 steps)
14 R1 = OAc, R2 = H
77% (2 steps)
AcO
15 R1 = H, R2 = OH
16 R1 = OH, R2 = H
HO
AcO
80%
AcO
21
HO
OH
AcO
22
OAc
OH
OAc
23
OH
OH
OH
OH
OH
O
OH
OH
HO
N
HO
N
O
8
OH
O
CuSO4 5H2O
sodium ascorbate
CH3OH
N
N
O
N
N
O
O
O
N
Cl
N
Cl
N
Cl
N
O
N
N
O
44%
(2 steps)
O
or
O
O
OCH3
OCH3
OCH3
56% from 15
4
1
2
65% from 16
N3
N3
OAc
CH3ONa
CH3OH
TMSN3, SnCl4
DCM
AcO
HO
O
O
AcO
O
Scheme 1 Synthesis of berberine derivatives 1 and 2
89%
AcO OAc
HO OH
26
AcO OAc
24
25
OH
HO
8
CuSO4 5H2O
sodium ascorbate
CH3OH
O
OH
N
N
O
O
Cl
N
N
49%
(2 steps)
O
OCH3
5
Scheme 2 Synthesis of berberine derivatives 3–5
CCK-8 assay for cytotoxicity
It has been proved that berberine and its derivatives have the
ability to up-regulate the reactive oxygen species (ROS), hence
depressing the proliferation of HepG2 cells.36 This feature can be
applied for the cytotoxicity assessment of berberine derivatives.
Therefore, all the synthesized berberine derivatives were tested for
their cytotoxicity through a standard CCK-8 assay37 at different
concentrations, and their half maximal inhibitory concentration
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 3
Please do not adjust margins

Please do not adjust margins
MedChemComm
Page 4 of 10
ARTICLE
Journal Name
(IC50) was calculated (Figure 5 and Table 1). The viability of HepG2 Table 1 The half maximal inhibitory concentratiVoienwsAr(tiIcCle50O)nlionef
DOI: 10.1039/C9MD00036D
cell obviously decreased with the concentration increasing from 0.2 berberine and its derivatives on HepG2 cells
to 625 μg/mL, which indicated that all the synthesized berberine
Compounds
1
R =
IC50 (μg/mL)
derivatives exerted cytotoxicity through a dose-dependent manner.
The glucose, galactose and mannose modified berberine derivatives
1–3 exhibited lower cytotoxicity than berberine, in particular
compound 3 with an IC50 value of 72.19, which was nearly 1.5 folds
of berberine (Figure 5A and Table1). While, the rhamnose and
ribose modified berberine derivatives 4 and 5 showed slightly
higher cytotoxicity than berberine (Figure 5B and Table1).
62.94 ± 0.032
2
3
59.29 ± 0.014
72.19 ± 0.051
4
49.27 ± 0.013
5
44.02 ± 0.017
53.91 ± 0.014
Berberine
–
Anti-diabetic investigation on HepG2 cell lines
HepG2 cell line is derived from human Hepatocellular carcinoma
and exhibits similar phenotype as hepatocyte.4 The insulin
receptors on surface of HepG2 cells can significantly decrease under
the stimulation of high level of insulin, and then results in low
glucose consumption (GC).38, 39 Consequently, HepG2 cell line has
been widely used as models for in vitro anti-diabetic investigation.
In order to screen the anti-diabetic activity of the synthesized
berberine derivatives 1–5, we determined the GC values of HepG2
cells19 after treatment with different concentrations (0.2, 1 and 5
μg/mL) of berberine and its derivatives (Figure 6). All the GC values
of berberine and its derivatives on HepG2 cell lines were higher
than that of the negative control group (0.5‰ DMSO), which
indicated that berberine and its derivatives had different degrees of
anti-diabetic activity. Compared with berberine, the carbohydrate
modified derivatives showed higher anti-diabetic activity, in
particular the mannose and ribose modified berberine derivatives 3
and 5 (Table 2). These two compounds showed the highest anti-
diabetic activity when the drug concentration was 5 μg/mL, and
induced an increase of GC values on HepG2 cell lines by 10.81% and
14.83%, respectively. More importantly, compounds 3 and 5 still
showed high anti-diabetic activity at concentrations of 0.2 and 1
μg/mL, suggesting their low effective concentrations. The galactose
modified berberine derivative 2 had similar activity as berberine at
high concentrations (1 and 5 μg/mL), but with poor activity at low
concentration (0.2 μg/mL). The rhamnose modified berberine
derivative 4 showed relatively poor anti-diabetic activity both at
high and low concentrations. Furthermore, modification of
berberine with glucose (compound 1) exhibited similar anti-diabetic
activity at both high and low concentrations.
Fig. 5 The effect of (A) glucose, galactose and mannose modified
berberine derivatives (1, 2 and 3), (B) rhamnose and ribose
modified berberine derivatives (4 and 5) on the HepG2 cell viability.
Data is expressed as means ± SD; n = 3.
4 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins

Page 5 of 10
MedChemComm
Please do not adjust margins
Journal Name
ARTICLE
View Article Online
DOI: 10.1039/C9MD00036D
Fig. 6 The effect of glucose, galactose, mannose, rhamnose and
ribose modified berberine derivatives (1, 2, 3, 4 and 5) on glucose
consumption of HepG2 cells after 24 h treatment. Data are
expressed as means ± SD; n = 6.
Fig. 7 Stability of berberine and compound 3 under acidic conditions.
Conclusions
Table 2 Data of glucose consumption activity in vitro for all
compounds with a concentration of 5 μg/mL (means ± SD, n = 8)
In conclusion, some novel carbohydrate modified berberine
derivatives were designed and synthesized. Synthesis of azido
sugars was employed the Carrington’s method, which could be
directly converted the acetylated sugar into azido sugar with high
efficiency and stereoselectivity. Attaching of the azido sugars to the
alkynylated berberine derivative 8 was finished by “click” chemistry,
and all the target compounds were achieved with high yields.
Therefore, this strategy could be applied for the convenient
synthesis of more complex carbohydrate modified berberine
derivatives. All the synthesized compounds were subjected to the
cytotoxicity measurement through the CCK-8 assay on HepG2 cell
lines, and the glucose, galactose and mannose modified berberine
derivatives 1–3 exhibited lower cytotoxicity than berberine.
Furthermore, their anti-diabetic activity was investigated on the
HepG2 cell line. The results indicated that the mannose and ribose
modified berberine derivatives 3 and 5 showed higher anti-diabetic
activity than berberine both at high and low concentrations.
Consequently, the mannose modified berberine derivative 3, with
low cytotoxicity, high anti-diabetic activity on the HepG2 cell lines
and high stability under acidc conditions could be potential
candidate for further research. To further investigate the structure-
activity relationship, it is interesting to prepare more berberine
derivatives with hydrophilic carbohydrate moiety, such as the
oligosaccharide-based berberine conjugates, and this work is now
under processing in our group.
Percentage increase in GC (%)
Compound
GC (Mm)
compared with berberine
5‰ DMSO
3.41 ± 0.033
4.72 ± 0.071
4.83 ± 0.053
4.71 ± 0.033
5.23 ± 0.025
4.55 ± 0.023
5.42 ± 0.028
/
berberine
/
1
2
3
4
5
2.23
0.21
10.81
–3.60
14.83
Stability under acidic conditions
Berberine derivative 3, with low cytotoxicity and high anti-
diabetic activity, was further subjected to stability investigation
under acidic conditions. Both berberine and compound 3 were
dissolved in the mix solution of acetonitrile and water (1/1, v/v, 0.5
mg/mL), which its pH value was adjuted to 1.5 using phosphoric
acid. The solution was then analyzed by HPLC with the absorption
wavelength at 320 nm (Shim-pack C8 250mm*4.6mm/5um column).
The elution was 50% acetonitrile in a 0.05 mol/L of aqueous KH2PO4,
and the flow rate was 1 mL/min. The results (Figure 5) indicated
that the content of berberine declined by 1.8% after 5 hours, while
that of derivative 3 only declined by 0.3%. Moreover, derivative 3
showed higher aqueous solubility (9 mg/mL) than berberine (1.75
mg/mL). The improvement on acid stability and aqueous solubility
of berberine derivatives have been proved critical for enhancing
their bio-availability.4, 21
Experimental procedures
General procedures
Berberine chloride was purchased from Tokyo Chemical Industry
(TCI, B0450). HepG2 cell lines were acquired from Shanghai Gefan
Biotechnology Co., Ltd (GF589). CCK-8 was obtained from Dōjindo
Laboratories (Dojindo). DMEM cell culture was bought from GE
Healthcare Life Sciences (SH30243.01). All other reagents were
commercial and used without further purification unless otherwise
noted. Thin layer chromatography (TLC) was performed with silica
gel HF254 plates and detected by charring with 30% (v/v) H2SO4 in
1
methanol or by a UV detector. H NMR and 13C NMR spectra were
recorded on a Bruker AV 400 spectrometer, and chemical shifts (δ)
are given in ppm downfield from internal TMS as an internal
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 5
Please do not adjust margins

Please do not adjust margins
MedChemComm
Page 6 of 10
ARTICLE
Journal Name
standard. High-resolution mass spectra (HRMS) were recorded on M in methanol) until the pH value reached 10.0. The reaction was
View Article Online
Agilent QTOF 6520 using electrospray ionization (ESI) technique to stirred at room temperature for 2 h, and TLDCO(CI:H102C.1l023/m9/Cet9hMaDn0o0l,053/61D,
introduce the sample.
V/V) showed the conversion of the starting material. To this mixture
was added 9-O-(propargyl) berberine chloride (8, 176 mg, 0.447
mmol) followed by CuSO4∙5H2O (67 mg, 0.268 mmol) and sodium
ascorbate (53 mg, 0.268 mmol). The reaction was then heated to 60
oC and stirred for 5 h. After TLC (CH2Cl2/methanol/water, 10/10/1,
V/V) showed the conversion of the starting material, the solvent
was evaporated, and the resultant residue was subjected to column
chromatography to give the desired 1 (150 mg, 56%) as a yellow
solid. 1H NMR (400 MHz, DMSO-d6): δ 9.59 (s, 1 H, CH=N), 8.92 (s, 1
H, ArH), 8.61 (s, 1 H, triazole-H), 8.22 (d, J = 9.2 Hz, 1 H, ArH), 8.04
(d, J = 9.2 Hz, 1 H, ArH), 7.78 (s, 1 H, ArH), 7.09 (s, 1 H, ArH), 6.18 (s,
2 H, –OCH2O–), 5.54 (d, J = 9.2 Hz, 1 H, glucose-H1), 5.50–5.43 (m, 2
H, CH2), 5.38 (s, 2 H, CH2), 5.20 (s, 1 H, OH), 4.89 (t, J = 15.2 Hz, 2 H,
CH2), 4.62 (s, 1 H, OH), 4.11 (s, 3 H, OCH3), 3.76 (t, J = 8.8 Hz, 1 H),
3.71–3.67 (m, 1 H), 3.45–3.41 (m, 2 H), 3.25–3.17 (m, 4 H); 13C NMR
(100 MHz, DMSO-d6): δ 151.40, 150.32, 148.14, 145.54, 142.89,
142.06, 137.94, 133.38, 131.17, 127.02, 124.97, 124.53, 122.42,
120.85, 120.74, 108.90, 105.91, 102.56, 87.95, 80.42, 77.34, 72.60,
70.12, 66.97, 61.22, 57.56, 55.88, 26.82; ESI-TOF HRMS (m/z): calcd
9-O-(propargyl) berberine chloride (8). Berberine chloride (5 g,
13.48 mmol) was heated at 190 oC under high vacuum (20–30
mmHg) for 30 min to give berberrubine 6 as a purple solid. The
crude solid was dissolved in 50 mL of DMF, and propargyl bromide
(7, 4.73 g, 40.44 mmol) was added. The reaction was heated to 80
oC and stirred for 6 h, and the color of the mixture was changed
from purple to yellow. Then, the reaction was cooled to room
temperature and stirred for another 2 h, the mixture was filtered,
the filter cake was washed with cold methanol and dried to give 9-
O-(propargyl) berberine chloride 8 (3.83 g, 72%) as a yellow solid. 1H
NMR (400 MHz, DMSO-d6): δ 9.88 (s, 1 H, CH=N), 8.97 (s, 1 H,
ArH),8.23 (d, J = 9.2 Hz, 1 H, ArH), 8.04 (d, J = 9.2 Hz, 1 H, ArH), 7.80
(s, 1 H, ArH), 7.10 (s, 1 H, ArH), 6.18 (s, 2 H, –OCH2O–), 5.10 (d, J =
2.4 Hz, 2 H, –CH2C≡CH), 4.96 (t, J = 6.4 Hz, 2 H, CH2), 4.08 (s, 3 H,
OCH3), 3.62–3.61 (m, 1 H, –CH2C≡CH), 3.21 (t, J = 6.0 Hz, 2 H, CH2);
13C NMR (100 MHz, DMSO-d6): δ 151.18, 150.36, 148.17, 145.79,
141.18, 138.11, 133.40, 131.20, 127.03, 124.73, 122.56, 120.87,
120.77, 108.90, 105.94, 102.58, 80.26, 79.24, 61.42, 57.63, 55.78,
+
for C28H29N4O9 , [M – Cl]+, 565.1929; found, 565.1936.
26.81; ESI-TOF HRMS (m/z): calcd for C22H18NO4 , [M – Cl]+,
+
360.1230; found, 360.1238.
2,3,4,6-Tetra-O-acetyl-β-D-galactopyranosyl azide (14). Compound
14 was achieved from 10 (5.0 g, 12.82 mmol) as a white solid (3.68
g, 77%), using the same procedure for the synthesis of 13. H NMR
1
2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyl azide (13). To the
solution of 1,2,3,4,6-penta-O-acetyl-D-glucopyranose 9 (5.0 g, 12.82
mmol) in dry CH2Cl2 (30 mL) was added hydrogen bromide (33% in
acetic acid, 15 mL) at 0 oC. The reaction was allowed to warm to
room temperature and stirred for 2 h, and then poured into 150 mL
of ice water. After which, the mixture was extracted with 150 mL of
CH2Cl2, the organic layer was washed with cold saturated NaHCO3
solution (50 mL × 2) followed by cold water (50 mL). The organic
phase was dried with anhydrous Na2SO4 and the solvent was
evaporated to give 11 (2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl
bromide) as a colorless oil. Next, the crude oil was dissolved in 30
(400 MHz, CDCl3): δ 5.42 (d, J = 3.2 Hz, 1 H, H-4), 5.17 (t, J = 9.2 Hz, 1
H, H-2), 5.05 (dd, J =3.6, 10.4 Hz, 1 H, H-3), 4.59 (d, J = 8.8 Hz, 1 H,
H-1), 4.21–4.13 (m, 2 H, 2 × H-6), 4.02 (t, J =6.4 Hz, 1 H, H-5), 2.17 (s,
3 H, CH3CO), 2.10 (s, 3 H, CH3CO), 2.07 (s, 3 H, CH3CO), 1.99 (s, 3 H,
CH3CO); 13C NMR (100 MHz, CDCl3): δ 170.34, 170.09, 169.97,
169.34, 88.33, 72.91, 70.75, 68.10, 66.87, 61.23, 20.67, 20.66,
20.62, 20.52; ESI-TOF HRMS (m/z): calcd for C14H19N3O9Na, [M +
Na]+, 396.1014; found, 396.1012; calcd for C14H23N4O9, [M + NH4]+,
391.1460; found, 391.1455.
mL of DMF, and a solution of NaN3 (2.5 g, 38.46 mmol) in 5 mL of 9-O-(β-D-galactopyranosyl-1H-1,2,3-triazole) berberine chloride
o
water was added at 0 C. The reaction was slowly warmed to room (2). Compound 2 was achieved from 14 (200 mg, 0.536 mmol) and 8
temperature and stirred for 3 h. After TLC (petroleum ether/ethyl (176 mg, 0.447 mmol) as a yellow solid (174 mg, 65%), using the
1
acetate, 2/1, V/V) showed the conversion of the starting material, same procedure for the synthesis of 1. H NMR (400 MHz, DMSO-
the reaction was diluted with 150 mL of CH2Cl2, and washed with d6): δ 9.62 (s, 1 H, CH=N), 8.88 (s, 1 H, ArH), 8.66 (s, 1 H, triazole-H),
water (100 mL × 3). The organic layer was dried and evaporated, 8.20 (d, J = 8.4 Hz, 1 H, ArH), 8.02 (d, J = 8.8 Hz, 1 H, ArH), 7.75 (s, 1
and the desired 13 (3.30 g, 69%) was obtained as a white crystals H, ArH), 7.07 (s, 1 H, ArH), 6.17 (s, 2 H, –OCH2O–), 5.52–5.26 (m, 4
1
after recrystallization from ethanol. H NMR (400 MHz, CDCl3): δ H, 2 × CH2), 4.95–4.68 (m, 4 H, galactose-H1, OH, CH2), 4.10 (s, 3 H,
5.22 (t, J = 9.6 Hz, 1 H, H-3), 5.11 (t, J = 9.6 Hz, 1 H, H-2), 4.96 (t, J = OCH3), 3.77 (s, 1 H), 3.68 (s, 1 H), 3.54–3.49 (m, 2 H), 3.26–3.17 (m,
9.2 Hz, 1 H, H-4), 4.64 (d, J = 8.8 Hz, 1 H, H-1), 4.27 (dd, J =4.8, 12.4 4 H); 13C NMR (100 MHz, DMSO-d6): δ 151.42, 150.31, 148.11,
Hz, 1 H, H-6), 4.18 (dd, J =2.0, 12.4 Hz, 1 H, H-6´), 3.80 (dq, J =2.4, 145.72, 142.88, 142.04, 137.89, 133.33, 131.23, 127.01, 124.94,
10.0 Hz, 1 H, H-5), 2.11 (s, 3 H, CH3CO), 2.08 (s, 3 H, CH3CO), 2.04 (s, 124.50, 122.46, 120.83, 120.70, 108.90, 105.91, 102.52, 88.52,
3 H, CH3CO), 2.01 (s, 3 H, CH3CO); 13C NMR (100 MHz, CDCl3): δ 78.83, 74.17, 69.60, 68.88, 60.85, 57.54, 55.85, 26.83; ESI-TOF
+
170.57, 170.09, 169.28, 169.18, 87.93, 74.06, 72.63, 70.67, 67.92, HRMS (m/z): calcd for C28H29N4O9 , [M – Cl]+, 565.1929; found,
61.68, 20.70, 20.56, 20.54; ESI-TOF HRMS (m/z): calcd for 565.1931.
C14H19N3O9Na, [M + Na]+, 396.1014; found, 396.1007; calcd for
C14H23N4O9, [M + NH4]+, 391.1460; found, 391.1453.
2,3,4,6-Tetra-O-acetyl-α-D-mannopyranosyl azide (19α). To the
solution of 1,2,3,4,6-penta-O-acetyl-D-mannopyranose 17 (2.0 g,
9-O-(β-D-glucopyranosyl-1H-1,2,3-triazole) berberine chloride (1). 5.13 mmol) in dry CH2Cl2 (10 mL) was added TMSN3 (1.35 mL, 10.26
To the solution of 2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl azide mmol) followed by SnCl4 (0.30 mL, 2.56 mmol) at 0 oC under
13 (200 mg, 0.536 mmol) in methanol (5 mL) was added CH3ONa (1 nitrogen atmosphere, the reaction was allowed to warm to room
6 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins

Page 7 of 10
MedChemComm
Please do not adjust margins
Journal Name
ARTICLE
temperature and stirred for
2 h. After TLC showed the 133.37, 131.12, 127.05, 125.64, 124.51, 122.45, 120.87, 120.76,
View Article Online
disappearance of the starting material, the mixture was diluted with 108.89, 105.91, 102.56, 86.50, 72.83, 72.D4O2,I:7101..14043,9/6C89.M81D,060603.862D,
CH2Cl2 (40 mL), and washed with saturated NaHCO3 solution (50 mL 57.54, 55.85, 26.83, 18.19; ESI-TOF HRMS (m/z): calcd for
+
× 3). The organic phase was dried with anhydrous Na2SO4 and the C28H29N4O8 , [M − Cl]+, 549.1980; found, 549.1986.
solvent was evaporated, the resultant crude oil was purified by
2,3,5-Tri-O-acetyl-β-D-ribofuranosyl azide (25). Compound 25 was
achieved from 24 (5.0 g, 15.72mmol) as a white solid (4.21 g, 89%),
using the same procedure for the synthesis of 19α. H NMR (400
MHz, CDCl3): δ 5.36 (d, J = 2.0 Hz, 1 H), 5.33 (dd, J = 4.8, 6.4 Hz, 1 H),
5.14 (dd, J = 2.0, 4.4 Hz, 1 H), 4.41 (dd, J = 3.2, 12.0 Hz, 1 H), 4.37–
4.34 (m, 1 H), 4.14 (dd, J = 4.0, 12.4 Hz, 1 H, H-5), 2.13 (s, 3 H,
CH3CO), 2.12 (s, 3 H, CH3CO), 2.08 (s, 3 H, CH3CO); 13C NMR (100
MHz, CDCl3): δ 170.55,169.52, 169.38, 92.69, 79.41, 74.51, 70.50,
63.03, 20.68, 20.51, 20.46; ESI-TOF HRMS (m/z): calcd for
C11H15N3O7Na, [M + Na]+, 324.0808; found, 324.0813.
column chromatography (petroleum ether/ethyl acetate, 2/1, V/V)
to give 19α (1.68 g, 88%) as an colorless oil. 1H NMR (400 MHz,
CDCl3): δ 5.39 (d, J = 1.2 Hz, 1 H, H-1), 5.32–5.23 (m, 2 H, H-3, H-4),
5.16 (t, J = 2.0 Hz, 1 H, H-2), 4.30 (dd, J = 5.6, 12.4 Hz, 1 H, H-6),
4.19–4.11 (m, 2 H, H-5, H-6´), 2.17 (s, 3 H, CH3CO), 2.12 (s, 3 H,
CH3CO), 2.06 (s, 3 H, CH3CO), 2.00 (s, 3 H, CH3CO); 13C NMR (100
MHz, CDCl3): δ 170.57, 169.84, 169.72, 169.61, 87.48, 70.67, 69.21,
68.26, 65.67, 62.17, 20.80, 20.70, 20.66, 20.59; ESI-TOF HRMS
1
(m/z): calcd for C14H19N3O9Na, [M
+
Na]+, 396.1014; found,
396.1011; calcd for C14H23N4O9, [M + NH4]+, 391.1460; found,
391.1458.
9-O-(β-D-ribofuranosyl-1H-1,2,3-triazole) berberine chloride (5).
Compound 5 was achieved from 25 (161 mg, 0.536 mmol) and 8
(176 mg, 0.447 mmol) as a yellow solid (150 mg, 49%), using the
9-O-(α-D-mannopyranosyl-1H-1,2,3-triazole) berberine chloride
(3). Compound 3 was achieved from 19α (200 mg, 0.536 mmol) and
8 (176 mg, 0.447 mmol) as a yellow solid (137 mg, 52%), using the
1
same procedure for the synthesis of 1. H NMR (400 MHz, DMSO-
1
d6): δ 9.65 (s, 1 H, CH=N), 8.93 (s, 1 H, ArH), 8.53 (s, 1 H, triazole-H),
8.24 (d, J = 8.4 Hz, 1 H, ArH), 8.03 (d, J = 8.8 Hz, 1 H, ArH), 7.79 (s, 1
H, ArH), 7.09 (s, 1 H, ArH), 6.18 (s, 2 H, –OCH2O–), 5.94 (d, J = 4.4 Hz,
1 H, H1), 5.56 (d, J = 6.0 Hz, 1 H), 5.47 (s, 2 H), 5.28 (d, J = 4.4 Hz, 1
H), 4.98 (t, J = 3.6 Hz, 1 H), 4.89 (t, J = 4.2 Hz, 2 H), 4.36–4.32 (m, 1
H), 4.10 (s, 3 H, CH3), 3.96 (s, 1 H), 3.61–3.56 (m, 1 H), 3.52–3.46 (m,
2 H), 3.19 (t, J = 6.4 Hz, 2 H); 13C NMR (100 MHz, DMSO-d6): δ
151.32, 150.34, 148.17, 145.58, 142.09, 137.96, 133.39, 131.13,
127.05, 124.48, 124.21, 122.38, 120.86, 120.76, 108.90, 105.91,
102.56, 92.55, 86.41, 75.69, 70.78, 67.07, 61.72, 57.53, 55.86,
same procedure for the synthesis of 1. H NMR (400 MHz, DMSO-
d6): δ 9.64 (s, 1 H, CH=N), 8.92 (s, 1 H, ArH), 8.45 (s, 1 H, triazole-H),
8.21 (d, J = 9.2 Hz, 1 H, ArH), 8.03 (d, J = 8.8 Hz, 1 H, ArH), 7.78 (s, 1
H, ArH), 7.09 (s, 1 H, ArH), 6.18 (s, 2 H, –OCH2O–), 5.90 (d, J = 3.2 Hz,
1 H, mannose-H1), 5.49 (s, 2 H, CH2), 5.27–5.23 (m, 1 H, OH), 5.07–
5.03 (m, 2 H, CH2), 4.89 (s, 2 H, CH2), 4.60 (t, J = 5.2 Hz, 1 H, OH),
4.39 (s, 1 H), 4.10 (s, 3 H, OCH3), 3.76 (s, 1 H), 3.60–3.50 (m, 3 H),
3.25–3.15 (m, 3 H); 13C NMR (100 MHz, DMSO-d6): δ 151.36, 150.32,
148.15, 145.55, 142.00, 137.98, 133.40, 131.12, 127.03, 125.41,
124.49, 122.40, 120.88, 120.77, 108.89, 105.92, 102.55, 86.16,
78.86, 71.64, 68.50, 68.00, 66.93, 61.09, 57.54, 55.87, 26.83; ESI-
+
26.83; ESI-TOF HRMS (m/z): calcd for C27H27N4O8 , [M − Cl]+,
+
TOF HRMS (m/z): calcd for C28H29N4O9 , [M – Cl]+, 565.1929; found,
535.1823; found, 535.1828.
565.1935.
Measurement of cytotoxicity by CCK-8 assay
2,3,4-Tri-O-acetyl-α-L-rhamnopyranosyl azide (22). Compound 22
was achieved from 21 (5.0 g, 15.06mmol) as a colorless oil (3.80 g,
80%), using the same procedure for the synthesis of 19α. H NMR
(400 MHz, CDCl3): δ 5.31 (d, J = 1.2 Hz, 1 H, H1), 5.21 (dd, J = 3.2, 10
Hz, 1 H), 5.15–5.14 (m, 1 H), 5.09 (t, J = 10 Hz, 1 H), 4.07–4.00 (m, 1
H), 2.16 (s, 3 H, CH3CO), 2.06 (s, 3 H, CH3CO), 1.99 (s, 3 H, CH3CO),
1.29 (d, J = 8.4 Hz, 3 H, CH3); 13C NMR (100 MHz, CDCl3): δ 169.92,
169.85, 169.82, 87.51, 70.50, 69.50, 68.63, 68.30, 20.83, 20.76,
20.64, 17.46; ESI-TOF HRMS (m/z): calcd for C12H17N3O7Na, [M +
Na]+, 338.0959; found, 338.0964.
HepG2 cells were maintained in DMEM cell medium containing 10%
heat-inactivated fetal bovine serum (FBS) and 100 μg/mL
penicillin/streptomycin in a humidified incubator (Thermo Scientific,
Series 8000 WJ) with 5% CO2 atmosphere at 37 ºC. Cells were
collected with 0.25% trypsin solution when the cell confluency was
approximately 80-90%. Then, the HepG2 cells were plated on 96-
well tissue culture plates with a density of 1×105 cells/mL. After 12
hours incubation, the media was removed and replaced with fresh
media. All the compounds were all dissolved in 5‰ DMSO-water
solution for cell treatment, the cells were treated with different
concentration of the compounds (0.2, 1.0, 5.0, 25, 125.0, 625
μg/mL) for 12 hours, and the negative control was incubated
without addition of any compounds. After that, 20 μL of CCK-8
solution (5 mg/mL) was added to each well and the cells were
cultured for 4 hours. The absorbance was measured by spectra MR
(DYNEX, Series1SPA-0093) at 570 nm. Furthermore, the absorbance
of different concentrations (0.2, 1.0, 5.0, 25, 125.0, 625 μg/mL) of
the compounds were also measured to eliminate the influence of
colors of drugs. The IC50 values were defined as the drug
concentration that inhibits 50% cell growth by setting the viability
of untreated cell as 100%.
1
9-O-(α-L-rhamnopyranosyl-1H-1,2,3-triazole) berberine chloride
(4). Compound 4 was achieved from 22 (169 mg, 0.536 mmol) and 8
(176 mg, 0.447 mmol) as a yellow solid (138 mg, 44%), using the
1
same procedure for the synthesis of 1. H NMR (400 MHz, DMSO-
d6): δ 9.65 (s, 1 H, CH=N), 8.92 (s, 1 H, ArH), 8.42 (s, 1 H, triazole-H),
8.22 (d, J = 9.2 Hz, 1 H, ArH), 8.00 (d, J = 9.2 Hz, 1 H, ArH), 7.78 (s, 1
H, ArH), 7.09 (s, 1 H, ArH), 6.17 (s, 2 H, –OCH2O–), 5.88 (d, J = 2.0 Hz,
1 H, rhamnopyranose-H1), 5.48 (s, 2 H, CH2), 5.26 (d, J = 4.8 Hz, 1
H), 5.02 (t, J = 5.6, 8.4 Hz, 2 H, CH2), 4.89 (t, J = 6.0 Hz, 2 H, CH2),
4.39–4.36 (m, 1 H), 4.10 (s, 3 H, CH3), 3.77–3.73 (m, 1 H), 3.19 (t, J =
6.4 Hz, 2 H), 1.10 (d, J = 6.4 Hz, 3 H, CH3); 13C NMR (100 MHz,
DMSO-d6): δ 151.43, 150.33, 148.16, 145.59, 141.89, 137.98,
Investigation of anti-diabetic activity on HepG2 cell lines
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 7
Please do not adjust margins

Please do not adjust margins
MedChemComm
Page 8 of 10
ARTICLE
Journal Name
HepG2 cells were suspended in the DMEM cell medium containing
10% heat-inactivated fetal bovine serum (FBS) and 100 μg/mL 12. G. Y. Pan, Z. J. Huang, G. J. Wang, J. PD. OFaI:w10ce.1t0t3,9X/.CD9M. LDi0u0, 0X3.6CD.
penicillin/streptomycin. Then, cells were seeded in 96-well plates Zhao, J. G. Sun and Y. Y. Xie, Planta Med., 2003, 69, 632-636.
with a density of 2×105 cells in each well, and cultured for 12 hours 13. Z. Cheng, T. Pang, M. Gu, A.-H. Gao, C.-M. Xie, J.-Y. Li, F.-J. Nan
in a humidified incubator at 37 °C with 5% CO2 atmosphere. and J. Li, Biochim. Biophys. Acta, 2006, 1760, 1682-1689.
Thereafter, all the cells were divided into four groups, including 14. J. A. Hawley and S. J. Lessard, Acta Physiol. (Oxf.), 2007, 192,
black control group, solvent control group (5‰ DMSO), berberine 127-135.
chloride group, and carbohydrate modified berberine derivatives 15. N. Turner, J.-Y. Li, A. Gosby, S. W. C. To, Z. Cheng, H. Miyoshi,
Ethnopharmacol., 2015, 161, 69-81.
View Article Online
group. Every berberine derivative was divided into three groups
M. M. Taketo, G. J. Cooney, E. W. Kraegen, D. E. James, L.-H.
based on dosage (0.2, 1.0, and 5.0 μg/mL), and the solvent control
Hu, J. Li and J.-M. Ye, Diabetes, 2008, 57, 1414-1418.
group and berberine chloride group has only dosage group (5.0 16. E. D. Rosen and O. A. MacDougald, Nat. Rev. Mol. Cell. Biol.,
μg/mL). After two days of incubation, the glucose concentrations of 2006, 7, 885-896.
each well were determined by glucose oxidase method.40 The 17. T. P. T. Pham, J. Kwon and J. Shin, Biochem. Biophys. Res.
values of glucose consumption (GC) were calculated by the glucose Commun., 2011, 413, 376-382.
concentrations of black wells subtracting the remaining glucose in 18. X. Xia, J. Yan, Y. Shen, K. Tang, J. Yin, Y. Zhang, D. Yang, H. Liang,
cell-treated wells.
J. Ye and J. Weng, PLOS ONE, 2011, 6, e16556.
19. S. Zhang, X. Wang, W. Yin, Z. Liu, M. Zhou, D. Xiao, Y. Liu and D.
Peng, Bioorg. Med. Chem. Lett., 2016, 26, 4799-4803.
20. Z. Chen, X. Ye, J. Yi, X. Chen and X. Li, Med. Chem. Res., 2012,
21, 1641-1646.
21. Z. Cheng, A.-F. Chen, F. Wu, L. Sheng, H.-K. Zhang, M. Gu, Y.-Y.
Li, L.-N. Zhang, L.-H. Hu, J.-Y. Li and J. Li, Bioorg. Med. Chem.,
2010, 18, 5915-5924.
22. Y.-Q. Shan, G. Ren, Y.-X. Wang, J. Pang, Z.-Y. Zhao, J. Yao, X.-F.
You, S.-Y. Si, D.-Q. Song, W.-J. Kong and J.-D. Jiang, Metabolism,
2013, 62, 446-456.
23. X. Qu, V. V. Khutoryanskiy, A. Stewart, S. Rahman, B.
Papahadjopoulos-Sternberg, C. Dufes, D. McCarthy, C. G.
Wilson, R. Lyons, K. C. Carter, A. Schätzlein and I. F. Uchegbu,
Biomacromolecules, 2006, 7, 3452-3459.
Conflict of Interest
The authors declare no competing interests.
Supplementary data
Electronic
supplementary
information
(ESI)
available:
Supplementary data associated with this article, including 1H and
the 13C NMR spectra, can be found in the online version, at
http://dx.doi.org/
Acknowledgments
This research was supported by grants from the Natural Science
Foundation of Shandong Province (No. ZR2018PB007) and Natural 24. S. A. W. Gruner, E. Locardi, E. Lohof and H. Kessler, Chem. Rev.,
Science Foundation of China (No. 81602982).
2002, 102, 491-514.
25. K. Wang, L. Chai, X. Feng, Z. Liu, H. Liu, L. Ding and F. Qiu, J.
Pharmaceut. Biomed., 2017, 139, 73-86.
References
26. J.-Y. Ma, R. Feng, X.-S. Tan, C. Ma, J.-W. Shou, J. Fu, M. Huang,
C.-Y. He, S.-N. Chen, Z.-X. Zhao, W.-Y. He, Y. Wang and J.-D.
Jiang, J. Pharm. Sci., 2013, 102, 4181-4192.
27. X.-P. He, Y.-L. Zeng, Y. Zang, J. Li, R. A. Field and G.-R. Chen,
Carbohydr. Res., 2016, 429, 1-22.
28. A. Shi, L. Huang, C. Lu, F. He and X. Li, Bioorg. Med. Chem.,
2011, 19, 2298-2305.
29. A. Ishiwata, H. Akao and Y. Ito, Org. Lett., 2006, 8, 5525-5528.
30. R. J. Williams, N. W. McGill, J. M. White and S. J. Williams, J.
Carbohyd. Chem., 2010, 29, 236-263.
1. A. Kumar, Ekavali, K. Chopra, M. Mukherjee, R. Pottabathini
and D. K. Dhull, Eur. J. Pharmacol., 2015, 761, 288-297.
2. M. Zhang and L. Chen, Acta Pharm. Sin. B, 2012, 2, 379-386.
3. J. Wang, T. Yang, H. Chen, Y.-N. Xu, L.-F. Yu, T. Liu, J. Tang, Z. Yi,
C.-G. Yang, W. Xue and F. Yang, Eur. J. Med. Chem., 2017, 127,
424-433.
4. Y. Ding, X. Ye, J. Zhu, X. Zhu, X. Li and B. Chen, J. Funct. Foods,
2014, 7, 229-237.
5. X. Bian, L. He and G. Yang, Bioorg. Med. Chem. Lett., 2006, 16,
1380-1383.
6. G. Liao, Z. Zhou and Z. Guo, Chem. Commun., 2015, 51, 9647-
9650.
7. J. Chen, T. Wang, S. Xu, A. Lin, H. Yao, W. Xie, Z. Zhu and J. Xu,
Eur. J. Med. Chem., 2017, 132, 173-183.
8. R. Gautam and S. M. Jachak, Med. Res. Rev., 2009, 29, 767-820.
9. M. Bahar, Y. Deng, X. Zhu, S. He, T. Pandharkar, M. E. Drew, A.
Navarro-Vázquez, C. Anklin, R. R. Gil, R. W. Doskotch, K. A.
Werbovetz and A. D. Kinghorn, Bioorg. Med. Chem. Lett., 2011,
21, 2606-2610.
31. B. Capon, Chem. Rev., 1969, 69, 407-498.
32. T. L. Lowary, J. Carbohyd. Chem., 2002, 21, 691-722.
33. L. Wang, S. Feng, S. Wang, H. Li, Z. Guo and G. Gu, J. Org.
Chem., 2017, 82, 12085-12096.
34. R. Carrington, G. Shaw and D. V. Wilson, J. Chem. Soc., 1965,
6864-6870.
35. L. Li, B. Lin, Z. Yang, L. Zhang and L. Zhang, Tetrahedron Lett.,
2008, 49, 4491-4493.
36. B. Liu, G. Wang, J. Yang, X. Pan, Z. Yang and L. Zang, PLOS ONE,
2011, 6, e21416.
10. H.-J. Maeng, H.-J. Yoo, I.-W. Kim, I.-S. Song, S.-J. Chung and C.-
K. Shim, J. Pharm. Sci., 2002, 91, 2614-2621.
11. J. Lan, Y. Zhao, F. Dong, Z. Yan, W. Zheng, J. Fan and G. Sun, J.
37. P. W. Burridge, Y. F. Li, E. Matsa, H. Wu, S.-G. Ong, A. Sharma,
A. Holmström, A. C. Chang, M. J. Coronado, A. D. Ebert, J. W.
Knowles, M. L. Telli, R. M. Witteles, H. M. Blau, D. Bernstein, R.
8 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins

Page 9 of 10
MedChemComm
Please do not adjust margins
Journal Name
ARTICLE
B. Altman and J. C. Wu, Nat. Med., 2016, 22, 547.
View Article Online
DOI: 10.1039/C9MD00036D
38. H. Zhang, J. Wei, R. Xue, J.-D. Wu, W. Zhao, Z.-Z. Wang, S.-K.
Wang, Z.-X. Zhou, D.-Q. Song, Y.-M. Wang, H.-N. Pan, W.-J.
Kong and J.-D. Jiang, Metabolism, 2010, 59, 285-292.
39. Q. Huang, L. Chen, H. Teng, H. Song, X. Wu and M. Xu, J. Funct.
Foods, 2015, 19, 487-494.
40. A. H. Kadish, R. L. Litle and J. C. Sternberg, Clin. Chem., 1968,
14, 116-131.
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 9
Please do not adjust margins

MedChemComm
Page 10 of 10
View Article Online
Novel carbohydrate modified berberine derivatives: Synthesis and in
DOI: 10.1039/C9MD00036D
vitro anti-diabetic investigation
Liwen Han,a,§ Wenlong Sheng,a,§ Xiaobin Li,a Sik Attila,b Houwen Lin,c Kechun Liu,a,* Lizhen Wang,a,*
higher anti-diabetic activity
than berberine
O
Carbohydrate
Cl
N3
Carbohydrate
N
N
O
N
N
Alkynylated
berberine
O
Cu catalyzed "click"
chemistry
OCH3
Carbohydrate modified
berberine derivatives 1-5
1