Synthesis and antitumor activity of new shikonin glycosides

Article abstract of DOI:10.1016/j.ejmech.2010.02.002

Eleven shikonin glycosides were synthesized and evaluated for their antitumor activity in vitro. Some of them were found to exhibit cytotoxic activities against both drug sensitive cell lines (K562, MCF-7 and HL60) and their drug resistant cell sublines (K562/ADR, MCF-7/ADR and HL60/ADR).

Full text of DOI:10.1016/j.ejmech.2010.02.002

European Journal of Medicinal Chemistry 45 (2010) 2713–2718
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Synthesis and antitumor activity of new shikonin glycosides
,
b
,
a
Yehua Su a, Jiansheng Xie b, Yanguang Wang a , Xun Hu , Xianfu Lin
*
*
a Department of Chemistry, Zhejiang University, 38# Zheda Road, Hangzhou 310027, PR China
b Cancer Institute, the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, PR China
a r t i c l e i n f o
a b s t r a c t
Article history:
Eleven shikonin glycosides were synthesized and evaluated for their antitumor activity in vitro. Some of
them were found to exhibit cytotoxic activities against both drug sensitive cell lines (K562, MCF-7 and
HL60) and their drug resistant cell sublines (K562/ADR, MCF-7/ADR and HL60/ADR).
Ó 2010 Published by Elsevier Masson SAS.
Received 25 October 2009
Received in revised form
28 January 2010
Accepted 1 February 2010
Available online 6 February 2010
Keywords:
Shikonin
Glycosides
Antitumor
Drug resistance
Synthesis
1. Introduction
2. Chemistry
Shikonin, one of the active components isolated from the
traditional medicinal herb Lithospermium erythrorhizon [1], has
attracted much attention due to its significant biological activi-
ties such as anti-inflammatory [2], antibacterial [3], wound
healing [4] and immunostimulatory activities [5]. It was also
reported that shikonin exhibited anticancer activity by inhibiting
telomerase and DNA topoisomerase I/II [6], and cancer cell
growth [7]. However, its poor toxicity and solubility prevent
shikonin from being an anticancer drug. Many efforts have been
made to synthesize new derivatives of shikonin, aiming at
discovering more effective antitumor drugs [8–10]. Previously,
we reported that shikonin could circumvent cancer drug resis-
tance by induction of a necroptotic death [11]. Encouraged by this
result, we are interested in the synthesis and evaluation of new
shikonin derivatives. We synthesized eleven shikonin glycosly
derivatives possessing acylglycosly at 10-OH and tested their in
vitro cytotoxicity on three pairs of drug sensitive and drug
resistant cell lines. These results showed all of the shikonin
acylglycosyl derivatives exhibited similar or stronger cytotoxicity
than shikonin and provided a good lead for the search of more
effective antitumor drug.
Shikonin was glycosylated at 10-OH to maintain the quinine
moiety, which has been identified as a pharmacophore that
commonly affords cytotoxicity [12]. Synthesis of acetyl-
shikonins is depicted in Scheme 1.
b-glycosyl
D
-Glucopyranose (1a),
D
-galactopyranose (1b),
-rhampyranose (1d), -xylopyranose (1e),
-arabofuranose (1g), -arabofuranose (1h), cellubiose
D
-mannopyr-
anose (1c),
anose (1f),
L
D
D-ribofur-
D
L
(1i), maltose (1j), and lactose (1k) were acetylated with acetic
anhydride and anhydrous sodium acetate to give the corresponding
acetylated sugars 2a–k. The anomeric O-acetyl groups of 2a–e
and 2i–k were selectively removed with hydrazine acetate to afford
3a–e and 3i–k [13], while the anomeric O-acetyl groups of fura-
noses 2f–h were removed with NaOCH3 in THF at 0 ꢀC to afford 3f–
h [14]. Treatment 3 with trichloroacetonitrile and potassium
carbonate yielded acetyl-a-glycosyl tirchloroacetimates 4 [15].
Compounds 4 as glycosyl donors reacted with shikonin under
BF3$EtO2 and 4 Å molecular sieve at nitrogen atmosphere to furnish
the corresponding shikonin acetyl-b-glycosides 5a–k according to
the published method [16]. The glycosylation of shikonin was
highly regioselective. The reaction occurred almost at the side chain
hydroxyl (10-OH) of shikonin, due to the hydrogen bond between
phenolic hydroxyl and quinoid carbonyl reducing the nucleophi-
licity of phenolic hydrogen.
All new products were well characterized by 1H NMR, 13C NMR
and IR spectroscopy as well as high-resolution mass spectrometry.
* Corresponding authors. Tel./fax: þ86 571 87951512.
E-mail addresses: orgwyg@zju.edu.cn (Y. Wang), huxun@zju.edu.cn (X. Hu).
0223-5234/$ – see front matter Ó 2010 Published by Elsevier Masson SAS.
doi:10.1016/j.ejmech.2010.02.002

2714
Y. Su et al. / European Journal of Medicinal Chemistry 45 (2010) 2713–2718
OH
O
5
3
6
7
4
1
2'
4'
1'
2
8
OH
5'
3'
OH
O
Shikonin
b (for 2a~e and 2i~k)
or c (for 2f~h)
O
O
O
2a~k
e
a
AcO
HO
AcO
OH
OH
OAc
3a~k
1a~k
OH O
OH O
O
R
O
AcO
d
R =
AcO
NH
CCl3
O
O
5a~k
4a~k
OAc
OAc
O
AcO
AcO
AcO
R
R
O
OAc
O
AcO
O
R
AcO
R
AcO
R
AcO
AcO
OAc
AcO
OAc
OAc
5d
5c
5a
5b
O
O
O
AcO
AcO
AcO
AcO
AcO
AcO
R
R
O
AcO
OAc
AcO
OAc
OAc
OAc
OAc
5e
5h
5f
5g
AcO
OAc
AcO
OAc
OAc
O
O
OAc
AcO
AcO
O
AcO
AcO
AcO
O
O
O
R
O
R
AcO
AcO
AcO
AcO
O
AcO
O
R
AcO
AcO
OAc
OAc
OAc
5i
5k
5j
Scheme 1. Synthesis of acetyl-b
-glycosyl shikonins 5a–k. Reagents and conditions: (a) Ac2O/AcONa, 80 ꢀC, 4 h, 90–98%; (b) AcOH.NH2NH2, THF, r.t., 4 h, 75%–90%; (c) NaOCH3, THF,
0
ꢀC, 30 min, 67–81%; (d) Cl3CCN, K2CO3, DCM, r.t., overnight, 60–85%; (e) BF3$EtO2, shikonin, 4 Å MS, DCM, ꢁ40 ꢀC, 1 h, 46–87%.
3. Results and discussions
transporters such as P-gp and MRP1, is one of the common mech-
anisms of cancer multidrug resistance. P-gp and MRP1 are trans-
membrane proteins and function as unilateral drug transporters
that expel intracellular drugs out of cells, so that intracellular drug
concentration may be kept at sublethal concentration. Not
less importantly, these drug transporters have broad substrate
specificity and thus can recognize many structurally and
functionally unrelated anticancer agents, leading to multidrug
In comparison to shikonin, the derivatives have similar or
stronger inhibitory activities against both drug sensitive cell lines
(K562, MCF-7 and HL60) and their drug resistant cell sublines:
K562/ADR, MCF-7/ADR and HL60/ADR (Table 1).
Cancer drug resistance is a major problem limiting cancer
chemotherapeutic efficacy [17]. Overexpression of drug
Table 1
In vitro cytotoxicity of shikonin derivatives.
Compounds
IC50(mM)
K562
K562/ADR (drug resistance factor)a
MCF-7
MCF-7/Adr (drug resistance factor)a
HL60
HL60/Adr (drug resistance factor)a
SH
5a
5b
5c
5d
5e
5f
5g
5h
5i
0.63
0.30
0.35
0.43
0.26
0.20
0.23
0.25
0.44
0.31
0.22
0.18
0.67 (1.06)
0.73 (2.38)
0.75 (2.12)
0.80 (1.86)
0.64 (2.46)
0.38 (1.63)
0.54 (2.35)
0.75 (3.00)
0.69 (1.57)
0.98 (3.14)
1.22 (5.54)
0.64 (3.53)
1.45
0.63
0.58
0.51
0.40
0.38
0.30
0.38
0.46
0.44
0.23
0.28
1.65 (1.14)
0.47 (0.75)
0.71 (1.24)
0.95 (1.86)
0.60 (1.50)
0.90 (2.40)
0.55 (1.83)
0.59 (1.55)
1.14 (2.48)
1.81 (4.16)
0.26 (1.13)
1.17 (4.18)
1.03
0.35
0.37
1.17
0.33
0.33
0.62
0.79
1.04
0.28
0.70
0.16
1.41 (1.37)
0.34 (0.99)
0.44 (1.18)
1.36 (1.16)
0.36 (1.09)
0.36 (1.11)
0.56 (0.90)
1.38 (1.75)
1.13 (1.09)
0.33 (1.16)
1.10 (1.57)
0.25 (1.52)
5j
5k
a
Drug resistance factor ¼ IC50 resistant cell line/IC50 sensitive cell line.

Y. Su et al. / European Journal of Medicinal Chemistry 45 (2010) 2713–2718
2715
resistance. K562/ADR and MCF-7/ADR are cell lines with charac-
teristics of P-gp overexpression, and HL60/ADR with MRP1 over-
expression. Since shikonin derivatives have similar cytotoxic
against both drug sensitive and resistant cells, i.e., overexpression
of P-gp and MRP1 does not affect their cellular cytotoxicity, these
compounds have potential to treat drug resistant cancer.
5.2.3. 1,2,3,4,6-Penta-O-acetylgalactopyranose (2c)
Yield 96%, white solid, mp: 134–136 ꢀC; 1H NMR (400 MHz,
CDCl3):
5.48 (d, J ¼ 8.4 Hz, 1H), 5.20 (s, 1H), 5.11 (t, J ¼ 9.6 Hz, 1H),
4.85 (dd, J ¼ 2.4, 10.0 Hz, 1H), 3.90 (m, 3H), 1.94 (s, 3H), 1.90 (s, 3H),
1.82 (s, 6H), 1.77 (s, 3H); 13C NMR (100 MHz, CDCl3):
170.2, 169.6,
d
d
169.4, 169.2, 167.7, 92.3, 70.2, 68.4, 68.0, 65.1, 61.7, 20.5, 20.4, 20.4,
20.3, 20.2.
4. Conclusion
5.2.4. 1,2,3,4-Tetra-O-acetylrhampyranose (2d)
In summary, a novel series of shikonin glycosides have been
synthesized and assessed for their preliminary anticancer activity
against both drug sensitive cell lines (K562, MCF-7 and HL60) and
their drug resistant cell sublines (K562/ADR, MCF-7/ADR and HL60/
ADR). Most of the shikonin derivatives exhibited good cytotoxicity
against all three cancer cell lines evaluated. The results suggest that
some of the synthesized glycosides, such as 5e and 5k, should be
potential compounds for treatment of certain drug resistant
cancers.
Yield 98%, Colorless oil; 1H NMR (400 MHz, CDCl3):
d 5.81 (s,1H),
5.44 (s, 1H), 5.05 (m, 2H), 3.65 (m, 1H), 2.18 (s, 3H), 2.08 (s, 3H), 2.03
(s, 3H), 1.98 (s, 3H), 1.26 (d, J ¼ 6.4 Hz, 3H); 13C NMR (100 MHz,
CDCl3):
d 170.2, 169.8, 168.7, 168.4, 90.2, 71.4, 70.6, 70.2, 68.4, 20.7,
20.7, 20.6, 20.4, 17.3.
5.2.5. 1,2,3,4-Tetra-O-acetylxylopyranose (2e)
Yield 95%, white solid, mp: 122–124 ꢀC; 1H NMR (400 MHz,
CDCl3):
d
5.70 (d, J ¼ 6.8 Hz,1H), 5.19 (t, J ¼ 8.0 Hz,1H), 5.00 (m, 2H),
4.13 (dd, J ¼ 4.8, 12.0 Hz, 1H), 3.51 (dd, J ¼ 8.0, 12.0 Hz, 1H), 2.09
(s,3H),2.04(s,3H),2.04(s,3H),2.03(s,3H);13CNMR(100 MHz,CDCl3):
5. Experimental
d
169.6, 169.1, 168.8, 91.9, 70.9, 69.4, 68.2, 62.6, 20.6, 20.5, 20.5, 20.4.
5.1. Chemistry
5.2.6. 1,3,4-Tetra-O-acetyl-
Yield 97%, white solid, mp: 109–111 ꢀC; 1H NMR (400 MHz,
CDCl3):
6.00 (d, J ¼ 4.0 Hz, 1H), 5.46 (d, J ¼ 2.8 Hz, 1H), 5.13 (m,
1H), 5.02 (d, J ¼ 2.8 Hz, 1H), 4.01 (m, 1H), 3.89 (m, 1H), 2.11 (s, 3H),
2.08 (m, 9H); 13C NMR (100 MHz, CDCl3):
169.8, 169.7, 169.4, 168.7,
90.8, 67.2, 66.0, 62.6, 20.8, 20.7, 20.6, 20.5.
D-ribofuranose (2f)
All reactions were performed under nitrogen atmosphere. The
reactions were monitored by TLC on silica gel GF254. Detection was
effected by examination under UV light and by sprayed with 20%
concd. H2SO4 in EtOH and heated at 110 ꢀC. Column chromatog-
raphy was performed on silica gel H. Intermediates and products
synthesized were characterized based on 1H NMR (bruker AM400)
and 13C NMR. CDCl3 was used as common solvent for the NMR.
Infrared (IR) spectra (KBr) were recorded on a Nicolet-470 Spec-
trometer. Melting points were determined by micro melting point
apparatus. Mass spectra (MS) were recorded on a Bruker Esruire
3000 Plus mass spectrometer through electron spray ionization.
High resolution mass spectral (HRMS) analyses were recorded on
BRUKER DALTONICS APEX III mass spectrometer using electron
spray ionization.
d
d
5.2.7. 1,2,3,4-Tetra-O-acetyl-
D-arabofuranose (2g)
Yield 90%, Colorless oil; 1H NMR (400 MHz, CDCl3):
d
5.68
(d, J ¼ 6.4 Hz, 1H), 5.32 (m, 2H), 5.13 (dd, J ¼ 3.6 8.8 Hz, 1H), 4.06
(dd, J ¼ 3.6 13.2 Hz, 1H), 3.79 (dd, J ¼ 1.2 13.2 Hz, 1H), 2.16 (s, 3H),
2.14 (s, 3H), 2.08 (s, 3H), 2.06 (s, 3H); 13C NMR (100 MHz, CDCl3):
d170.1,
169.8, 169.3, 169.0, 92.0, 69.8, 67.9, 67.0, 63.8, 20.8, 20.7, 20.6, 20.5.
5.2.8. 1,2,3,4-Tetra-O-acetyl-
Yield 91%, white solid, mp: 89–90 ꢀC; 1H NMR (400 MHz,
CDCl3):
L-arabofuranose (2h)
5.2. General method for the synthesis of compounds 2a–k
d
5.64 (d, J ¼ 6.8 Hz, 1H), 5.25 (m, 2H), 5.08 (dd, J ¼ 3.2
8.8 Hz, 1H), 4.01 (dd, J ¼ 3.6 12.8 Hz, 1H), 3.74 (d, J ¼ 13.2 Hz, 1H),
To glucose (10 mmol) and anhydrous sodium acetate (13 mmol)
was added acetic anhydride (100 mmol). The mixture was stirred at
80 ꢀC for 4 h (reaction progress was monitored by TLC). After cooled
to room temperature, the reaction solution was pounded into cold
saturated sodium bicarbonate solution. The mixture was stirred for
1.5 h, extracted with dichloromethane and dried over sodium
sulfate. The solvent was removed in vacuum and the residue was
purified by flash silica gel column chromatography (AcOEt/petro-
leum ether ¼ 1:4, v/v).
2.11 (s, 3H), 2.09 (s, 3H), 2.04 (s, 3H), 2.01 (s, 3H); 13C NMR
(100 MHz, CDCl3):
d 170.1, 169.8, 169.3, 169.0, 92.1, 69.8, 68.0, 67.1,
63.8, 20.8, 20.7, 20.6, 20.5.
5.2.9. 1,2,3,6,20,30,40,60-Octa-O-acetylmaltose (2i)
Yield 98%, white solid, mp: 163–165 ꢀC; 1H NMR (400 MHz,
CDCl3):
d
5.73 (d, J ¼ 7.6 Hz, 1H), 5.40 (d, J ¼ 3.6 Hz, 1H), 5.34
(t, J ¼ 5.6 Hz, 1H), 5.29 (t, J ¼ 8.8 Hz, 1H), 5.05 (t, J ¼ 10.0 Hz, 1H),
4.97 (t, J ¼ 8.8 Hz, 1H), 4.85 (dd, J ¼ 4.0, 10.4 Hz, 1H), 4.44 (d,
J ¼ 10.6 Hz, 1H), 4.22 (m, 2H), 4.04 (m, 2H), 3.92 (d, J ¼ 10.8 Hz, 1 H),
3.83 (d, J ¼ 9.6 Hz, 1H), 2.13 (s, 3H), 2.09 (s, 6H), 2.04 (s, 3H), 2.02
5.2.1. 1,2,3,4,6-Penta-O-acetylglucopyranose (2a)
Yield 93%, white solid, mp: 93–95 ꢀC; 1H NMR (400 MHz,
(s, 3H), 2.01 (s, 6H), 2.00 (s, 3H); 13C NMR (100 MHz, CDCl3):
d 170.5,
CDCl3):
d
5.72 (d, J ¼ 8.0 Hz, 1H), 5.26 (t, J ¼ 9.6 Hz, 1H), 5.14 (m, 2H),
170.4,170.3,170.0,169.8, 169.5,169.3, 168.7, 95.6, 91.1, 75.1, 72.8, 72.2,
70.8, 69.9, 69.2, 68.4, 67.8, 62.4, 61.3, 20.8, 20.7, 20.6, 20.5, 20.4.
4.30 (dd, J ¼ 4.4, 12.4 Hz, 1H), 4.11 (dd, J ¼ 1.6, 12.4 Hz, 1H), 3.85
(m, 1H), 2.12 (s, 3H), 2.09 (s, 3H), 2.04 (s, 6H), 2.02 (s, 3H); 13C NMR
(100 MHz, CDCl3): d 170.6, 170.0, 169.3, 169.2, 168.9, 91.6, 72.7, 72.6,
70.1, 67.6, 61.3, 20.8, 20.7, 20.5.
5.2.10. 1,2,3,6,20,30,40,60-Octa-O-acetylcellubiose (2j)
Yield 96%, white solid, mp: 188–190 ꢀC; 1H NMR (400 MHz,
5.2.2. 1,2,3,4,6-Penta-O-acetylmannopyranose (2b)
CDCl3):
d
5.64 (d, J ¼ 8.0 Hz,1H), 5.21 (t, J ¼ 5.2 Hz,1H), 5.04 (m, 3H),
Yield 98%, white solid, mp: 111–113 ꢀC; 1H NMR (400 MHz,
4.90 (t, J ¼ 8.0 Hz, 1H), 4.48 (m, 2H), 4.34 (dd, J ¼ 4.4, 12.0 Hz, 1H),
4.10 (dd, J ¼ 4.4, 12.0 Hz, 1H), 4.03 (d, J ¼ 11.2 Hz, 1H), 3.80
(t, J ¼ 9.6 Hz, 1H), 3.66 (m, 2H), 2.16 (s, 3H), 2.11 (s, 3H), 2.10 (s, 3H),
2.01 (m, 9H), 1.99 (s, 3H), 1.96 (s, 3H); 13C NMR (100 MHz, CDCl3):
CDCl3):
(dd, J ¼ 4.8, 12.4 Hz, 1H), 3.97 (m, 2H), 2.06 (s, 3H), 2.05 (s, 3H), 1.97
(s, 6H), 1.94 (s, 3H), 1.89 (s, 3H); 13C NMR (100 MHz, CDCl3):
170.2,
d
5.96 (m, 1H), 5.22 (d, J ¼ 6.4 Hz, 2H), 5.13 (s, 1H), 4.16
d
170.0, 169.4, 169.2, 168.8, 92.0, 71.5, 70.6, 67.6, 66.6, 60.9, 20.7, 20.5,
20.5, 20.4.
d
170.3, 170.1, 170.0, 169.5, 169.3, 169.1, 168.8, 168.7, 100.5, 91.4, 75.7,
73.4, 72.7, 72.2, 71.8, 71.4, 70.3, 67.7, 61.4, 20.7, 20.6, 20.5, 20.4, 20.3.

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Y. Su et al. / European Journal of Medicinal Chemistry 45 (2010) 2713–2718
5.2.11. 1,2,3,6,20,30,40,60-Octa-O-acetyllactose (2k)
2.05 (s, 3H), 2.04 (s, 3H), 2.00 (s, 3H); 13C NMR (100 MHz, CDCl3):
d 169.8, 169.7, 160.9, 93.1, 90.7, 69.8, 69.3, 68.5, 60.7, 20.6, 20.5 20.4.
Yield 93%, white solid, mp: 130–132 ꢀC; 1H NMR (400 MHz,
CDCl3):
d
5.67 (d, J ¼ 8.4 Hz, 1H), 5.35 (d, J ¼ 3.2 Hz, 1H), 5.25
(d, J ¼ 9.2 Hz, 1H), 5.05 (m, 3H), 4.47 (m, 2H), 4.12 (m, 3H), 3.85
(m, 3H), 2.16 (s, 3H), 2.12 (s, 3H), 2.10 (s, 3H), 2.07 (s, 3H), 2.06
(s, 3H), 2.05 (s, 3H), 2.04 (s, 3H), 1.97 (s, 3H); 13C NMR (100 MHz,
5.3.6. 2,3,6,20,30,40,60-Hepta-O-acetyl-
a-maltosyl trichloro-
acetimidate (4i)
Yield 66%, white solid, mp: 79–81 ꢀC; 1H NMR (400 MHz,
CDCl3):
d
170.3, 170.2, 170.1, 170.0, 169.6, 169.5, 168.9, 168.8, 100.9,
CDCl3):
d
8.65 (s, 1H), 6.45 (d, J ¼ 4.0 Hz, 1H), 5.58 (t, J ¼ 9.6 Hz, 1H),
91.4, 75.6, 73.4, 72.5, 70.8, 70.6, 70.4, 68.8, 66.5, 61.6, 60.7, 20.9,
20.8, 20.7, 20.6, 20.6, 20.5, 20.4.
5.42 (d, J ¼ 4.4 Hz, 1H), 5.36 (t, J ¼ 10.0 Hz, 1H), 5.04 (t, J ¼ 9.6 Hz,
1H), 4.98 (dd, J ¼ 3.6,10.0 Hz,1H), 4.85 (dd, J ¼ 3.6,10.8 Hz,1H), 4.46
(d, J ¼ 12.0 Hz, 1H), 4.21 (m, 3H), 4.04 (m, 2H), 3.92 (d, J ¼ 10.0 Hz,
1H), 2.10 (s, 3H), 2.07 (s, 3H), 2.04 (s, 3H), 2.01 (s, 3H), 2.00 (s, 3H),
5.3. General method for the synthesis of compounds 4a–e and 4i–k
1.98 (s, 3H), 1.96 (s, 3H); 13C NMR (100 MHz, CDCl3):
d 170.5, 170.4,
A solution of 2 (5 mmol) and hydrazine acetate (5.5 mmol) in
THF (20 mL) was stirred for 4 h at room temperature. After
removed the solvent, dichloromethane (40 mL) was added to
dissolve the residue. The organic phase was washed with water
(10 mL), dried over anhydrous sodium sulfate. The solvent was
removed in vacuum to give 3. K2CO3 (15 mmol) and tri-
chloroacetonitrile (12 mmol) was added to the solution of 3
(3 mmol) in anhydrate dichloromethane (40 mL). The mixture was
stirred at room temperature overnight. The resulting mixture was
filtered to give the organic phase. The organic phase was concen-
trated in vacuum and the residue was purified by flash silica gel
column chromatography (AcOEt/petroleum ether ¼ 1:4, v/v).
170.2,169.8,169.5,169.3,160.7, 95.6, 92.6, 90.5, 72.0, 71.8, 70.4, 70.0,
69.8, 69.2, 68.4, 67.8, 62.1, 61.2, 20.8, 20.6, 20.5, 20.4, 20.3, 20.2.
5.3.7. 2,3,6,20,30,40,60-Hepta-O-acetyl-
a-cellubiosyl trichloro-
acetimidate (4j)
Yield 60%, white solid, mp: 111–113 ꢀC; 1H NMR (400 MHz,
CDCl3):
d
8.66 (s, 1H), 6.48 (d, J ¼ 2.8 Hz, 1H), 5.53 (t, J ¼ 6.4 Hz, 1H),
5.08 (m, 3H), 4.93 (t, J ¼ 8.4 Hz, 1H), 4.54 (t, J ¼ 8.4 Hz, 2H), 4.39
(dd, J ¼ 4.0, 12.0 Hz, 1H), 4.11 (m, 3H), 3.84 (t, J ¼ 10.0 Hz, 1H), 3.67
(d, J ¼ 9.2 Hz, 1H), 2.11 (s, 3H), 2.09 (s, 3H), 2.04 (s, 3H), 2.03 (s, 3H),
2.01 (s, 6H), 1.98 (s, 3H); 13C NMR (100 MHz, CDCl3):
d 170.4, 170.2,
170.1, 170.0, 169.4, 169.2, 169.0, 160.9, 100.9, 92.8, 76.0, 73.0, 72.0,
71.0, 71.6, 70.9, 69.8, 69.2, 61.5, 61.3, 20.7, 20.6, 20.5, 20.4, 20.3.
5.3.1. 2,3,4,6-Tetra-O-acetyl-a-glucopyranosyl trichloro-
acetimidate (4a)
5.3.8. 2,3,6,20,30,40,60-Hepta-O-acetyl-
a-lactosyl trichloroacet-
Yield 70%, white solid, mp: 66–68 ꢀC; 1H NMR (400 MHz,
imidate (4k)
CDCl3):
d
8.69 (s, 1H), 6.56 (d, J ¼ 3.6 Hz, 1H), 5.57 (t, J ¼ 9.6 Hz, 1H),
Yield 69%, white solid, mp: 153–155 ꢀC; 1H NMR (400 MHz,
5.19 (t, J ¼ 9.6 Hz, 1H), 5.13 (dd, J ¼ 3.6, 10.0 Hz, 1H), 4.28 (dd, J ¼ 4.0,
12.4 Hz, 1H), 4.22 (m, 1H), 4.12 (dd, J ¼ 2.0, 12.4 Hz, 1H), 2.08 (s, 3H),
2.05 (s, 3H), 2.03 (s, 3H), 2.02 (s, 3H); 13C NMR (100 MHz, CDCl3):
CDCl3):
d
8.66 (s, 1H), 6.46 (d, J ¼ 3.6 Hz, 1H), 5.59 (t, J ¼ 9.6 Hz, 1H),
5.42 (d, J ¼ 4.4 Hz,1H), 5.37 (t, J ¼ 9.6 Hz, 1H), 5.06 (t, J ¼ 9.6 Hz, 1H),
5.01 (dd, J ¼ 3.6, 9.6 Hz, 1H), 4.86 (dd, J ¼ 4.0, 10.4 Hz, 1H), 4.48
(d, J ¼ 13.2 Hz, 1H), 4.23 (m, 3H), 4.05 (t, J ¼ 10.0 Hz, 2H), 3.93
(d, J ¼ 9.6 Hz, 1H), 2.11 (s, 3H), 2.08 (s, 3H), 2.05 (s, 3H), 2.02 (s, 3H),
2.00 (s, 3H), 1.99 (s, 3H), 1.97 (s, 3H); 13C NMR (100 MHz, CDCl3):
d
170.5, 170.0, 169.8, 169.4, 160.7, 92.8, 90.6, 69.9, 69.8, 69.6, 67.7,
61.3, 20.6, 20.5, 20.4.
5.3.2. 2,3,4,6-Tetra-O-acetyl-
a-mannopyranosyl trichloro-
d170.5,170.4,170.3,169.9,169.6,169.4,160.8, 95.7, 92.7, 90.6, 72.1, 71.9,
acetimidate (4b)
70.5, 70.1, 69.9, 69.3, 68.5, 67.9, 62.2, 61.3, 20.8, 20.7, 20.6, 20.5, 20.3.
Yield 62%, white solid, mp: 62–64 ꢀC; 1H NMR (400 MHz,
CDCl3):
(m, 1H), 4.16 (m, 2H), 2.19 (s, 3H), 2.07 (s, 3H), 2.05 (s, 3H), 2.00
(s, 3H); 13C NMR (100 MHz, CDCl3):
170.5, 169.8, 169.6, 169.5,
159.6, 94.4, 90.4, 71.1, 68.7, 67.7, 65.2, 61.9, 20.7, 20.6 20.5.
d
8.78 (s, 1H), 6.27 (s, 1H), 5.46 (s, 1H), 5.39 (m, 2H), 4.26
5.4. General method for the synthesis of compounds 4f–h
d
A mixture of 2f–h (5 mmol) and NaOMe (10 mmol) inTHF (20 mL)
was stirred at 0 ꢀC for 30 min and then neutralized with AcOH
(10 mmol). After removed the solvent in vacuum, dichloromethane
(40 ml) was added to dissolve the residue. The organic phase was
washed with water (10 mL) and dried over anhydrous sodium sulfate.
The solvent was removed in vacuum to give 3f–h. K2CO3 (15 mmol)
and trichloroacetonitrile (12 mmol) was added tothe solution of 3f–h
(3 mmol) in anhydrate dichloromethane (20 mL). The mixture was
stirred at room temperature overnight. The resulting mixture was
filtered to give the organic phase. After removed the solvent in
vacuum, the residue was purified by flash silica gel column chroma-
tography (AcOEt/petroleum ether ¼ 1:4, v/v).
5.3.3. 2,3,4,6-Tetra-O-acetyl-a-galactopyranosyl trichloro-
acetimidate (4c)
Yield 68%, white solid, mp: 112–114 ꢀC; 1H NMR (400 MHz,
CDCl3):
d
8.66 (s, 1H), 6.59 (d, J ¼ 3.2 Hz, 1H), 5.55 (d, J ¼ 2.8 Hz, 1H),
5.37 (m, 2H), 4.43 (t, J ¼ 6.8 Hz, 1H), 4.15 (dd, J ¼ 2.4, 11.2 Hz, 1H),
4.07 (dd, J ¼ 6.8, 11.2 Hz, 1H), 2.16 (s, 3H), 2.02 (s, 3H), 2.01 (s, 3H),
2.00 (s, 3H); 13C NMR (100 MHz, CDCl3):
d 170.2, 170.0, 170.0, 169.9,
160.8, 93.4, 90.6, 68.9, 67.4, 67.2, 66.8, 61.2, 20.6, 20.5, 20.5, 20.4.
5.3.4. 2,3,4-Tri-O-acetyl-a-rhampyranosyl trichloroacet-imidate
(4d)
5.4.1. 2,3,5-Tri-O-acetyl-
a
-
D
-ribofuranosyl trichloro-acetimidate (4f)
Yield 66%, white solid, mp: 78–80 ꢀC; 1H NMR (400 MHz,
Yield 49%, Colorless oil; 1H NMR (400 MHz, CDCl3):
d
8.74 (s,1H),
CDCl3):
d
8.74 (s, 1H), 6.21 (s, 1H), 5.47 (d, J ¼ 2.0 Hz, 1H), 5.37 (dd,
6.22 (d, J ¼ 3.6 Hz,1H), 5.46 (t, J ¼ 3.6 Hz,1H), 5.26 (t, J ¼ 3.6 Hz,1H),
5.22 (d, J ¼ 3.6 Hz, 1H), 5.17 (dd, J ¼ 2.8 12.8 Hz, 1H), 3.97 (dd, J ¼ 4.4
12.8 Hz, 1H), 2.12 (s, 3H), 2.11 (s, 3H), 2.05 (s, 3H); 13C NMR
J ¼ 10.4, 4.0 Hz, 1H), 5.18 (d, J ¼ 10.4 Hz, 1H), 4.10 (m, 1H), 2.19
(s, 3H), 2.08 (s, 3H), 2.01 (s, 3H), 1.28 (d, J ¼ 6 Hz, 3H); 13C NMR
(100 MHz, CDCl3):
d
169.9, 169.8, 169.7, 159.9, 94.6, 90.6, 70.2, 69.2,
(100 MHz, CDCl3):
d 169.9, 169.6, 169.5, 160.2, 95.1, 90.5, 66.6, 66.2,
68.8, 68.1, 20.8, 20.7 20.6, 17.4.
65.6, 63.2, 20.8, 20.6, 20.5.
5.3.5. 2,3,4-Tri-O-acetyl-
Yield 59%, white solid, mp: 96–97 ꢀC; 1H NMR (400 MHz,
CDCl3):
8.65 (s, 1H), 6.47 (d, J ¼ 3.2 Hz, 1H), 5.56 (t, J ¼ 5.6 Hz, 1H),
5.07 (m, 2H), 3.98 (dd, J ¼ 6.4, 10.8 Hz, 1H), 3.80 (t, J ¼ 6.8 Hz, 1H),
a
-xylopyranosyl trichloroacet-imidate (4e)
5.4.2. 2,3,5-Tri-O-acetyl-a-D-arabofuranosyl trichloro-acetimidate
(4g)
d
Yield 46%, white solid, mp: 130–132 ꢀC; 1H NMR (400 MHz,
CDCl3):
d
8.61 (s, 1H), 6.52 (d, J ¼ 2.8 Hz, 1H), 5.38 (m, 3H), 4.12

Y. Su et al. / European Journal of Medicinal Chemistry 45 (2010) 2713–2718
2717
(d, J ¼ 13.2 Hz, 1H), 3.84 (d, J ¼ 13.2 Hz, 1H), 2.13 (s, 3H), 2.00 (s, 3H),
1.99 (s, 3H); 13C NMR (100 MHz, CDCl3):
170.1, 170.0, 169.8, 160.9,
94.0, 90.8, 68.2, 67.1, 67.0, 62.7, 20.8, 20.6, 20.4.
5.5.4. 1-(1-(5,8-Dihydroxy-1,4-naphthoquinone-2-yl)-4-methyl-
d
3-pentenyloxy)-2,3,4-tri-O-acetyl-b-rhampyranoside (5d)
Yield 82%, red solid, mp: 117–118 ꢀC; 1H NMR (400 MHz, CDCl3):
12.60 (s, 1H), 12.48 (s, 1H), 7.19 (s, 2H), 7.17 (s, 1H), 5.26 (m, 2H),
d
5.4.3. 2,3,5-Tri-O-acetyl-
a
-
L
-arabofuranosyl trichloro-acetimidate
5.11 (t, J ¼ 3.2 Hz, 1H), 5.03 (m, 2H), 4.93 (s, 1H), 3.73 (m, 1H), 2.47
(4h)
(m, 2H), 2.15 (s, 3H), 2.04 (s, 3H), 2.00 (s, 3H), 1.67 (s, 3H), 1.58
Yield 57%, white solid, mp: 106–107 ꢀC; 1H NMR (400 MHz,
(s, 3H), 1.03 (d, J ¼ 3.2 Hz, 3H); 13C NMR (100 MHz, CDCl3):
d 178.4,
CDCl3):
d
8.72 (s, 1H), 5.95 (d, J ¼ 4.4 Hz, 1H), 5.31 (m, 2H), 5.22
176.9, 169.8, 169.8, 169.7, 167.1, 166.6, 149.1, 136.2, 132.7, 132.4, 132.1,
117.4, 111.6, 111.4, 97.4, 73.1, 70.6, 69.5, 68.8, 67.1, 33.2, 25.5, 20.7,
20.6, 20.6, 17.8, 17.1; HRMS (ESI) Calcd for [M þ Na]þ ¼ 583.1786,
(dd, J ¼ 3.2 6.4 Hz, 1H), 4.14 (dd, J ¼ 7.2 12.0 Hz, 1H), 3.80 (dd, J ¼ 3.2
12.0 Hz, 1H), 2.16 (m, 6H), 2.01 (s, 3H); 13C NMR (100 MHz, CDCl3):
d
169.8, 169.7, 168.9, 160.5, 94.4, 90.6, 68.0, 67.8, 65.5, 61.0, 20.7,
found: [M þ Na]þ ¼ 583.1796; IR (KBr)
n
(cmꢁ1) ¼ 3465, 1741, 1605,
20.6, 20.5.
1453, 1230, 1076.
5.5. General method for the synthesis of compounds 5a–k
5.5.5. 1-(1-(5,8-Dihydroxy-1,4-naphthoquinone-2-yl)-4-methyl-
3-pentenyloxy)-2,3,4-tri-O-acetyl-b-xylopyranoside (5e)
To the mixture of 4 (1 mmol), shikonin (1 mmol) and 4 Å
molecular sieve (0.4 g) in dichloromethane (5 mL) was added
dropwise a solution of BF3$Et2O (0.1 mmol) in dichloromethane
(3 mL) at ꢁ40 ꢀC. After another hour of stirring, Et3N (1 mmol) was
added to the mixture, and AcOH (1 mmol) was added. The reaction
mixture was filtered to give the organic phase. Removed the solvent
of organic phase in vacuum and the residue was purified by flash
chromatography to afford 5a–k (51–87%).
Yield 86%, red solid, mp: 153–155 ꢀC; 1H NMR (400 MHz, CDCl3):
12.57 (s, 1H), 12.48 (s, 1H), 7.18 (m, 3H), 5.12 (m, 2H), 5.00 (m, 1H),
4.93 (m, 2H), 4.67 (d, J ¼ 6.4 Hz, 1H),3.96 (dd, J ¼ 11.6, 4.4 Hz), 3.27
(dd, J ¼ 10.4, 9.6 Hz,1H), 2.52 (m,1H), 2.33 (m,1H), 2.09 (s, 3H), 2.08
(s, 3H), 2.03 (s, 3H), 1.68 (s, 3H), 1.56 (s, 3H); 13C NMR (100 MHz,
d
CDCl3):
d 180.5, 179.2 170.5, 170.2, 169.8, 166.3, 165.8, 150.4, 135.8,
133.6, 132.9, 132.4, 118.6, 112.3, 112.0, 101.2, 75.4, 71.6, 71.2, 69.0,
62.4, 34.1, 26.3, 21.2, 21.2, 21.1, 18.4; HRMS (ESI) Calcd
for [M þ Na]þ ¼ 569.1629, found: [M þ Na]þ ¼ 569.1619; IR (KBr)
5.5.1. 1-(1-(5,8-Dihydroxy-1,4-naphthoquinone-2-yl)-4-methyl-
n
(cmꢁ1) ¼ 3447, 1747, 1605, 1451, 1229, 1078, 1058.
3-pentenyloxy)-2,3,4,6-tetra-O-acetyl-b-glucopyranoside (5a)
Yield 86%, red solid, mp: 58–60 ꢀC; 1H NMR (400 MHz, CDCl3):
12.49(s, 1H), 12.41 (s, 1H), 7.12 (m, 3H), 5.13 (m, 1H), 5.00 (m, 3H),
5.5.6. 1-(1-(5,8-Dihydroxy-1,4-naphthoquinone-2-yl)-4-methyl -
d
3-pentenyloxy)-2,3,4-tri-O-acetyl-b-D-ribofuranoside (5f)
4.86 (m, 1H), 4.60 (d, J ¼ 8.0 Hz, 1H), 3.95 (m, 2H), 3.53 (m, 1H), 2.45
Yield 51%, red solid, mp: 70–72 ꢀC; 1H NMR (400 MHz, CDCl3):
d12.59 (s, 1H), 12.47 (s, 1H), 7.19 (m, 2H), 7.15 (m, 1H), 5.44
(m, 1H), 2.26 (m, 1H), 2.00 (s, 3H), 1.95 (s, 3H), 1.94 (s, 3H), 1.85
(s, 3H), 1.62 (s, 3H), 1.49 (s, 3H); 13C NMR (100 MHz, CDCl3):
d
180.4,
(t, J ¼ 3.2 Hz, 1H), 5.13 (m, 1H), 5.08 (m, 1H), 5.05 (m, 1H), 4.99
(m, 2H), 3.81 (dd, J ¼ 3.2, 12.4 Hz, 1H), 3.66 (dd, J ¼ 5.6, 12.4 Hz,
1H), 2.53 (m, 1H), 2.41 (m, 1H), 2.12 (s, 3H), 2.09 (s, 3H), 2.07
179.2, 170.3, 170.2, 169.3, 169.1, 165.3, 164.8, 149.7, 135.2, 133.6,
132.3, 131.7, 118.1, 111.8, 111.5, 100.8, 75.6, 72.6, 71.8, 71.4, 68.2, 61.5,
33.7, 25.7, 20.6, 20.5, 20.4, 20.3, 17.9; HRMS (ESI) Calcd
for [M þ Na]þ ¼ 641.1841, found: [M þ Na]þ ¼ 641.1864; IR (KBr)
(s, 3H), 1.68 (s, 3H), 1.57 (s, 3H); 13C NMR (100 MHz, CDCl3):
d179.2,
177.8, 169.8, 169.7, 169.5, 166.5, 166.0, 149.5, 135.8, 132.7, 132.6,
132.2, 117.9, 111.8, 111.5, 98.5, 74.0, 68.4, 66.5, 66.4, 61.6, 33.5, 25.7,
20.8, 20.8, 20.7, 18.0; HRMS (ESI) Calcd for [M þ Na]þ ¼ 569.1629,
n
(cmꢁ1) ¼ 3467, 1750, 1611, 1455, 1229, 1042.
5.5.2. 1-(1-(5,8-Dihydroxy-1,4-naphthoquinone-2-yl)-4-methyl-3-
found: [M þ Na]þ ¼ 569.1639; IR (KBr)
n
(cmꢁ1) ¼ 3466, 1751, 1612,
pentenyloxy)-2,3,4,6-tetra-O-acetyl-
b-manno-pyranoside (5b)
1455, 1226, 1078.
Yield 46%, red solid, mp: 113–115 ꢀC; 1H NMR (400 MHz, CDCl3):
12.57 (s, 1H), 12.46 (s, 1H), 7.18 (m, 3H), 5.30 (m, 2H), 5.23 (t,
d
5.5.7. 1-(1-(5,8-Dihydroxy-1,4-naphthoquinone-2-yl)-4-methyl-
J ¼ 10 Hz, 1H), 5.12 (t, J ¼ 6.8 Hz, 1H), 5.04 (m, 2H), 4.14 (dd, J ¼ 5.6,
12.0 Hz, 1H), 3.89 (dd, J ¼ 1.6, 12.4 Hz, 1H), 3.83 (m, 1H), 2.51 (m,
1H), 2.45 (m, 1H), 2.14 (s, 3H), 2.02 (s, 3H), 2.00 (s, 3H), 1.94 (s, 3H),
3-pentenyloxy)-2,3,4-tri-O-acetyl-b-D-arabo-furanoside (5g)
Yield 63%, red solid, mp: 52–53 ꢀC; 1H NMR (400 MHz, CDCl3):
12.59 (s, 1H), 12.43 (s, 1H), 7.17 (m, 2H), 7.04 (s, 1H), 5.24 (m, 2H),
d
1.67 (s, 3H), 1.58 (s, 3H); 13C NMR (100 MHz, CDCl3):
d
177.9, 176.4,
5.16 (m, 2H), 5.01 (dd, J ¼ 3.2, 8.8 Hz, 1H), 4.35 (d, J ¼ 6.4 Hz1 H),
4.04 (dd, J ¼ 3.6, 12.8 Hz, 1H), 3.58 (dd, J ¼ 1.6, 12.8 Hz, 1H), 2.57 (m,
1H), 2.46 (m, 1H), 2.16 (s, 3H), 2.13 (s, 3H), 2.03 (s, 3H), 1.66 (s, 3H),
170.4, 169.9, 169.7, 169.7, 167.8, 167.3, 148.9, 136.6, 133.1, 132.7, 132.1,
117.4, 111.7, 111.5, 97.5, 73.2, 69.4, 69.3, 68.9, 66.0, 62.4, 33.5, 25.7,
20.8, 20.7, 20.6, 20.4, 18.0; HRMS (ESI) Calcd for
1.50 (s, 3H); 13C NMR (100 MHz, CDCl3):
d 176.4, 175.6, 170.3, 170.0,
[M þ Na]þ ¼ 641.1841, found: [M þ Na]þ ¼ 641.1853; IR (KBr)
n
169.4, 169.1, 168.6, 148.3, 135.2, 133.5, 133.1, 132.1, 118.1, 111.8, 111.5,
98.9, 72.9, 70.0, 69.3, 67.3, 62.8, 34.3, 25.7, 21.0, 20.8, 20.6, 18.0;
(cmꢁ1) ¼ 3483, 1758, 1611, 1451, 1215, 1053.
HRMS
(ESI)
Calcd
for
[M þ Na]þ ¼ 569.1629,
found:
5.5.3. 1-(1-(5,8-Dihydroxy-1,4-naphthoquinone-2-yl)-4-methyl-
[M þ Na]þ ¼ 569.1635; IR (KBr)
n
(cmꢁ1) ¼ 3472, 1749, 1611, 1455,
3-pentenyloxy)-2,3,4,6-tetra-O-acetyl-
b-galacto-pyranoside (5c)
1223, 1061.
Yield 87%, red solid, mp: 61–63 ꢀC; 1H NMR (400 MHz, CDCl3):
12.58 (s, 1H), 12.51 (s, 1H), 7.21 (m, 3H), 5.36 (m, 1H), 5.27 (dd,
d
5.5.8. 1-(1-(5,8-Dihydroxy-1,4-naphthoquinone-2-yl)-4-methyl-
3-pentenyloxy)-2,3,4-tri-O-acetyl- -arabofuranoside (5h)
Yield 75%, red solid, mp: 127–129 ꢀC; 1H NMR (400 MHz,
CDCl3): 12.57 (s, 1H), 12.50 (s, 1H), 7.25 (s, 1H), 7.19 (m, 2H), 5.22
J ¼ 10.4, 8.0 Hz, 1H), 5.14 (t, J ¼ 6.8 Hz, 1H), 5.02 (dd, J ¼ 3.2, 6.4 Hz,
1H), 4.97 (dd, J ¼ 4.0, 6.8 Hz, 1H), 4.62 (d, J ¼ 7.6 Hz, 1H), 3.99
(d, J ¼ 6.4 Hz, 2H), 3.83 (t, J ¼ 6.4 Hz, 1H), 2.52 (m, 1H), 2.35
(m, 1H), 2.17 (s, 3H), 2.09 (s, 3H), 2.00 (s, 3H), 1.92 (s, 3H), 1.69
b-L
d
(m, 2H), 5.12 (t, J ¼ 6.8 Hz, 1H), 5.09 (dd, J ¼ 3.6, 8.8 Hz, 1H), 4.97
(dd, J ¼ 4, 5.6 Hz, 1H), 4.61 (d, J ¼ 6.4 Hz, 1H), 3.86 (dd, J ¼ 3.6,
12.8 Hz,1H), 3.52 (d, J ¼ 12.8 Hz,1H), 2.53 (m,1H), 2.36 (m,1H), 2.13
(s, 3H), 2.10 (s, 3H), 2.07 (s, 3H), 1.68 (s, 3H), 1.55 (s, 3H); 13C NMR
(s, 3H), 1.56 (s, 3H); 13C NMR (100 MHz, CDCl3):
d 180.4, 179.2,
170.2, 170.1, 170.0, 169.1, 165.1, 164.6, 149.5, 135.1, 133.6, 132.1,
131.6, 117.9, 111.6, 111.4, 101.1, 75.3, 70.7, 70.6, 68.7, 66.7, 61.0, 33.4,
25.7, 20.6, 20.5, 20.4, 20.3, 17.8; HRMS (ESI) Calcd
for [M þ Na]þ ¼ 641.1841, found: [M þ Na]þ ¼ 641.1856; IR (KBr)
(100 MHz, CDCl3):
d 180.5, 179.1, 170.3, 170.1, 169.3, 165.4, 164.9,
150.0, 135.4, 133.4, 132.2, 131.8, 118.1, 111.8, 111.6, 100.5, 74.6, 69.8,
69.2, 67.2, 62.7, 33.5, 25.8, 20.9, 20.8, 20.7, 18.0; HRMS (ESI) Calcd
n
(cmꢁ1) ¼ 3468, 1753, 1612, 1455, 1221, 1079.

2718
Y. Su et al. / European Journal of Medicinal Chemistry 45 (2010) 2713–2718
for [M þ Na]þ ¼ 569.1629, found: [M þ Na]þ ¼ 569.1614; IR (KBr)
5.6. In vitro cytotoxic activity
n
(cmꢁ1) ¼ 3468, 1747, 1609, 1454, 1226, 1061.
5.6.1. Cell lines and cellular proliferation assay
5.5.9. 1-(1-(5,8-Dihydroxy-1,4-naphthoquinone-2-yl)-4-methyl-
MCF-7, HL60 and K562 cells were maintained in RPMI 1640
containing 10% fetal bovine serum. MCF-7/ADR, HL60/ADR and
K562/ADR cells were grown in RPMI 1640 containing 10% fetal
3-pentenyloxy)-2,3,6,20,30,40,60-hepta-O-acetyl-
b-maltoside (5i)
Yield 79%, red solid, mp: 72–74 ꢀC; 1H NMR (400 MHz, CDCl3):
d
12.59 (s,1H),12.43 (s,1H), 7.17 (m, 3H), 5.52 (d, J ¼ 5.2 Hz,1H), 5.49
bovine serum and 1 mg/mL doxorubicin. All compounds were
(d, J ¼ 4 Hz, 1H), 5.36 (t, J ¼ 10.0 Hz, 1H), 5.15 (m, 1H), 5.03
(t, J ¼ 6.0 Hz, 1H), 4.98 (s, 2H), 4.84 (dd, J ¼ 4.0, 10.4 Hz, 1H), 4.22
(m, 4H), 4.00 (m 2H), 3.80 (t, J ¼ 6.4 Hz, 1H), 3.58 (d, J ¼ 8.4 Hz, 1H),
2.36 (m,1H), 2.28 (m, 1H), 2.09 (s, 3H), 2.08 (s, 3H), 2.06 (s, 3H), 2.04
(s, 3H), 2.02 (s, 3H), 2.00 (s, 3H),1.98 (s, 3H),1.66 (s, 3H),1.56 (s, 3H);
dissolved in DMSO with the stock concentration of 20 mM, and
diluted with medium freshly before drug administration. MCF-7
and MCF-7/ADR cells were seeded into 96-well flat bottom
plates at density of 3000 cells/well. Twenty-four h after seeding,
each compound was added in triplicate. HL60 and HL60/ADR,
K562 and K562/ADR cells were seeded at density of 8000 and
3000 cells/well respectively. Compound was added after seeding
in triplicate. Cells in 96-well plate were incubated at 37 ꢀC in
13C NMR (100 MHz, CDCl3):
169.2, 169.1, 168.5, 150.8, 135.5, 133.5, 133.1, 131.4, 121.9, 118.9, 111.8,
111.4, 96.9, 94.7, 73.0, 72.2, 70.2, 69.7, 68.6, 68.2, 68.2, 68.1, 67.6,
63.7, 61.7, 35.1, 25.6, 21.4, 20.8, 20.8, 20.7, 20.6, 20.6, 20.5, 17.9;
d 176.7, 175.1, 170.5, 170.3, 170.1, 169.4,
a humidified atmosphere containing 5% CO2. After 72 h, 20
(5 mg/mL) was added into each well for 4 h incubation. After that,
the supernatant was removed and 150 L of dimethyl-sulphoxide
mL MTT
HRMS
(ESI)
Calcd
for
[M þ Na]þ ¼ 929.2686,
found:
[M þ Na]þ ¼ 929.2685; IR (KBr)
n
(cmꢁ1) ¼ 3468, 1750, 1612, 1455,
m
1228, 1040.
(DMSO, Sigma) was added into each well in order to dissolve
the blue-purple crystals of formazan. The absorbance was then
measured using a model ELX800 Micro Plate Reader (Bio-Tek
Instruments, Inc.) at 570 nm.
5.5.10. 1-(1-(5,8-Dihydroxy-1,4-naphthoquinone-2-yl)-4-methyl-
3-pentenyloxy)-2,3,6,20,30,40,60-hepta-O-acetyl-
b-cellubioside (5j)
Yield 85%, red solid, mp: 161–163 ꢀC; 1H NMR (400 MHz, CDCl3):
12.59(s, 1H), 12.41 (s, 1H), 7.18 (m, 2H), 7.02 (s, 1H), 5.06 (m, 6H),
d
Acknowledgments
4.93 (m, 1H), 4.53 (m, 2H), 4.38 (m, 2H), 4.13 (dd, J ¼ 4.8, 11.6 Hz,
1H), 4.05 (d, J ¼ 12.4 Hz, 1H), 3.82 (t, J ¼ 9.2 Hz, 1H), 3.66 (d,
J ¼ 8.8 Hz, 1H), 3.54 (dd, J ¼ 3.6, 9.6 Hz, 1H), 2.53 (m, 1H), 2.40 (m,
1H), 2.16 (s, 3H), 2.12 (s, 3H), 2.09 (s, 3H), 2.02 (s, 6H), 2.01 (s, 3H),
1.98 (s, 3H), 1.66 (s, 3H), 1.51(s, 3H); 13C NMR (100 MHz, CDCl3):
We thank China National 863 project (2007AA02Z143) for
financial support.
References
d
175.9, 175.0, 170.4, 170.1, 169.7, 169.6, 169.5, 169.2, 169.1, 168.9,
148.2, 135.0, 133.7, 133.3, 131.9, 118.3, 111.8, 111.5, 100.7, 99.0, 76.3,
73.7, 72.9, 72.7, 72.6, 71.9, 71.6, 71.5, 67.8, 61.6, 61.5, 34.2, 25.6, 20.8,
20.7, 20.6, 20.5, 17.9; HRMS (ESI) Calcd for [M þ Na]þ ¼ 929.2686,
[1] V.P. Papageorgiou, A.N. Assimopoulou, E.A. Couladouros, D. Hepworth,
K.C. Nicolaou, Angew. Chem. Int. Ed. 38 (1999) 270–301.
[2] (a) W.J. Wang, J.Y. Bai, D.P. Liu, L.M. Xue, X.Y. Zhu, Yaoxue Xuebao 29 (1994)
161–165;
(b) S. Tanaka, M. Tajima, M. Tsukada, M. Tabata, J. Nat. Prod. 49 (1986) 466–469.
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found: [M þ Na]þ ¼ 929.2690; IR (KBr)
n
(cmꢁ1) ¼ 3483, 1752, 1612,
1229, 1042.
5.5.11. 1-(1-(5,8-Dihydroxy-1,4-naphthoquinone-2-yl)-4-methyl
-3-pentenyloxy)-2,3,6,20,30,40,60-hepta-O-acetyl-
b-lactoside (5k)
Yield 81%, red solid, mp: 94–96 ꢀC; 1H NMR (400 MHz, CDCl3):
(b) Z.F. Plyta, T. Li, V.P. Papageorgiou, A.S. Mellidis, A.N. Assimopoulou,
E.N. Pitsinos, E.A. Couladouros, Bioorg. Med. Lett. 8 (1988) 3385–3390.
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[10] F. Yang, Y. Chen, W.H. Duan, C. Zhang, H. Zhu, J. Ding, Int. J. Cancer 119 (2006)
1184–1193.
d
12.57 (s,1H),12.42 (s,1H), 7.15 (m, 3H), 5.57 (d, J ¼ 5.2 Hz,1H), 5.45
(m,1H), 5.34 (d, J ¼ 2.8 Hz,1H), 5.14 (m, 2H), 5.06 (dd, J ¼ 3.6, 7.6 Hz,
1H), 4.96 (dd, J ¼ 3.6, 10.4 Hz, 1H), 4.56 (d, J ¼ 8 Hz, 1H), 4.26
(dd, J ¼ 2.4, 4.4 Hz, 1H), 4.20 (dd, J ¼ 2.0, 12.0 Hz, 1H), 4.07 (m, 3H),
3.90 (t, J ¼ 6.8 Hz, 1H), 3.77 (m, 1H), 3.60 (d, J ¼ 9.6 Hz, 1H), 2.36
(m, 1H), 2.29 (m, 1H), 2.14 (s, 3H), 2.08 (s, 3H), 2.07 (s, 3H), 2.02
(s, 3H), 2.00 (s, 3H), 1.94(s, 3H), 1.66 (s, 3H), 1.58(s, 3H), 1.55 (s, 3H);
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13C NMR (100 MHz, CDCl3):
d 177.5, 175.9, 170.6, 170.3, 170.2, 170.0,
169.3, 168.9, 168.4, 167.8, 150.7, 135.1, 133.2, 132.8, 132.0, 121.6, 119.2,
111.8,111.6,102.4, 96.9, 77.4, 73.2, 70.9, 70.8, 69.8, 68.8, 68.5, 67.1, 66.7,
63.4, 60.9, 35.3, 29.6, 25.7, 21.6, 20.8, 20.7, 20.6, 20.6, 20.5,17.9; HRMS
(ESI) Calcd for [M þ Na]þ ¼ 929.2986, found: [M þ Na]þ ¼ 929.2949;
[13] G. Excoffier, D. Gagnaire, J.P. Utille, Carbohydr. Res. 39 (1975) 368–373.
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(1986) 241–246.
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[16] R.R. Schmidt, J. Michel, Tetrahedron Lett 25 (1984) 821–824.
[17] W.D. Han, L. Li, X. Hu, Autophagy 3 (2007) 490–492.
IR (KBr)
n
(cmꢁ1) ¼ 3466, 1751, 1612, 1227, 1050.