Synthesis, structure characterization, and biological evaluation of some new 1,2,3-benzotriazole derivatives

Article abstract of DOI:10.1007/s00044-009-9243-3

Ten novel benzotriazole compounds were synthesized. Their chemical structures were confirmed by ¹H NMR, IR, and elemental analyses, coupled with three selected single-crystal structures (compounds A2, B3, and B5). Their antimycotic and antitumor activities were also investigated. The title compounds showed some antitumor activities, especially in the case of A3 and A4, which showed the most potent activity of propagation inhibition in liver and galactophore cancer cells. Birkhaueser Boston 2009.

Full text of DOI:10.1007/s00044-009-9243-3

MEDICINAL
CHEMISTRY
RESEARCH
Med Chem Res (2010) 19:970–983
DOI 10.1007/s00044-009-9243-3
ORIGINAL RESEARCH
Synthesis, structure characterization, and biological
evaluation of some new 1,2,3-benzotriazole derivatives
•
•
•
•
Jun Wan Xia Yan Cuiping Ma Sai Bi
Hai-Liang Zhu
Received: 17 February 2009 / Accepted: 25 August 2009 / Published online: 30 September 2009
¨
Ó Birkhauser Boston 2009
Abstract Ten novel benzotriazole compounds were synthesized. Their chemical
1
structures were confirmed by H NMR, IR, and elemental analyses, coupled with
three selected single-crystal structures (compounds A2, B3, and B5). Their anti-
mycotic and antitumor activities were also investigated. The title compounds
showed some antitumor activities, especially in the case of A3 and A4, which
showed the most potent activity of propagation inhibition in liver and galactophore
cancer cells.
Keywords Benzotriazole compounds Á Antimycotic activities Á
Antitumor activities Á Crystal structure
Introduction
Heterocyclic compounds containing nitrogen are considered to be the most effective
anticancer drugs used either as single agents or in combination for cancer therapy
(Lin et al., 2005; Fang et al., 2006; Wang et al., 2006). Some azole compounds,
especially triazole or benzimidazole derivatives, have been reported to have
cytotoxic potency on human cancer cells, e.g., breast carcinoma and liver cancer
J. Wan Á H.-L. Zhu (&)
State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing,
Jiangsu 210093, People’s Republic of China
e-mail: zhuhl@nju.edu.cn
J. Wan
e-mail: wanjundz@sohu.com
J. Wan Á X. Yan Á C. Ma Á S. Bi
Key Laboratory of Eco-Chemical Engineering, Ministry of Education, and College of Chemistry
and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042,
People’s Republic of China

Med Chem Res (2010) 19:970–983
971
(Sau¨czewski et al., 2004; Pagliai et al., 2006; Leonetti et al., 2004; Saberi et al.,
2006). During recent years studies have shown that several benzotriazole
compounds can act to diminish inflammation, as antivirus, antifungal, and
antimumor agents, and as selective inhibitors of PTP1B and antidepressants,
resulting from the potent bioactivity of benzotriazole (Xu et al., 2003; Emami et al.,
2004; Biagi et al., 2001; Scapin et al., 2003; Antonio et al., 2004). Moreover,
Touami et al. (1997) also reported that the novel conjugates of benzotriazole
photonucleases and DNA minor groove binders exhibit enhanced cleavage
efficiency and unique cleavage selectivity (Touami et al., 1997). Al-Soud et al.
(2003) reported that some triazole derivations showed remarkable activity against
leukemia, ovarian, renal, and lung cancers McClure et al. (2005) also reported that
benzotriazoles are significantly more potent inhibitors applied to p38 MAP kinase
experimentally than benzoimidazolones. The X-ray crystal structure of benzotria-
zole derivatives also shows that the triazole group as the H-bond acceptor, but
unexpectedly as a dual acceptor, inducing movement of the crossover connection of
p38R, was the most useful in ranking potency (McClure et al., 2005). And niacin
has long been used for the treatment of lipid disorders and for the prevention of
atherosclerosis, as a result of its ability to raise high-density lipoprotein (HDL)
levels (Semple et al., 2006). The design and synthesis of new benzotriazole
compounds with higher physiological and biological properties have been the focus
of recent studies in our laboratories (Wan et al., 2006; Zhang et al., 2007a).
Exploiting the concept of bioisosterism (Cui and Nan, 2006), a series of novel
title compounds including benzotriazole was synthesized. Single crystals suitable
for X-ray measurements of three compounds were also obtained by slow
evaporation of mixed solvents at room temperature. Furthermore, their biologi-
cal and antitumor activities were investigated. The results show that the
compounds 3-(1H-benzo[d][1,2,3]triazol-1-yl)-1-oxo-1-p-tolylpropan-2-yl nicotin-
ate and 3-(1H-benzo[d][1,2,3] triazol-1-yl)-1-(4-chlorophenyl)-1-oxopropan-2-yl
nicotinate possess potent propagation inhibition activity in liver and galactophore
cancer cells, respectively, and the toxicity of the compounds has a time-quantity
efficiency relationship.
Experimental
General comments
All reagents were obtained from commercial suppliers and were used without
further purification. Solvents for reactions, extraction, and chromatography were of
analytical grade. Intermediates I and II were readily prepared according to the
reported methods (Wan et al., 2006; Abonia et al., 2004). To a 250-m flask were
added 0.02 mol of intermediate II in 50 ml of acetic and 0.05 mol of sodium
acetate. Then bromine, 0.02 mol (3.2 g), was added dropwise, with stirring, at room
temperature. The reaction was maintained until the mixture became colorless for
about 4 h. Then 50 ml of water and 20 ml of chloroform were added. The organic
layer was successively washed with saturated sodium bicarbonate solution and

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Med Chem Res (2010) 19:970–983
brine, then dried over anhydrous magnesium sulfate to get III. Melting points were
measured on a Yanaco MP-500 melting-point apparatus and were uncorrected.
Elemental analyses were measured with a Vario EL III analyzer. IR spectra (4000–
400 cm-1), as KBr pellets, were recorded on a Nicolet FT-IR 510P spectropho-
tometer. 1H NMR was recorded on a 500-MHz JEOL FX-90Q NMR spectrometer in
CDCl3 as the solvent and with TMS as an internal standard. Column chromatog-
raphy was carried out on silica gel (unactivated, neutral, 100–200 mesh).
Preparation of A1–A6, B1, and B3–B5
Triethylamine (2.8 ml, 0.02 mol) was added dropwise, with stirring, to a solution of
intermediate III (0.02 mol) and nicotinic acid or isonicotinic acid (0.02 mol) in
acetone at 0°C. The mixture was stirred for another 5 h in an ice-water bath, then
filtered, and the filtrate was concentrated to give a crude product which was poured
into a saturated sodium chloride solution, followed by extraction with chloroform.
The combined organic extracts were washed with water, dried with MgSO4, and
filtered. Removal of the solvent gave the residue, which was chromatographed on
silica or recrystallized to get the desired products of A1–A6, B1, and B3–B5 in pure
form. Similarly, the single crystals were obtained by slow evaporation of mixed
solvents at room temperature over a period of several days.
3-(1H-Benzo[d][1,2,3]triazol-1-yl)-1-oxo-1-phenylpropan-2-yl nicotinate (A1)
Purified by recrystallization from petroleum ether/DMF (5:1, v/v). Yield, 37.8%;
yellow solid; m.p., 145.8–147.0°C. Anal. Calcd. for C21H16N4O3 (%): C 67.73, H
4.33, N 15.05. Found (%): C 67.53, H 4.43, N 15.34. mmax/cm-1 (KBr): 3133 (=C–
1
H), 1750, 1710 (s, C=O), 1585 (s, C=N), 1102, 1227 (s, C–O). H NMR (CDCl3):
5.18 and 5.31(2H, CH2), 6.70 (1H, CH), 7.26–9.07 (13H, phenyl H).
3-(1H-Benzo[d][1,2,3]triazol-1-yl)-1-(4-methoxyphenyl)-1-oxopropan-2-yl
nicotinate (A2)
Purified by column chromatography using petroleum ether/ethyl acetate (3:1, v/v) as
the eluent; the single crystal suitable for X-ray measurements was obtained by slow
evaporation of petroleum ether and alcohol (1:2, v/v) at room temperature over a
period of several days. Yield, 35.1%; yellow solid; m.p., 160.1–161.8°C. Anal.
Calcd. for C22H18N4O4 (%): C 65.66, H 4.51, N 13.92. Found (%): C 65.86, H 4.35,
N 13.88., mmax/cm-1 (KBr): 3130(=C–H), 1750, 1710 (s, C=O), 1585 (s, C=N),
1
1102, 1227 (s, C–O)., H NMR (CDCl3): 3.90 (3H, methoxyl, CH3), 5.18 and 5.31
(2H, CH2), 6.68 (1H, CH), 6.99–9.08 (12H, phenyl H). ESI–MS m/z 403.1 (MH?).
3-(1H-Benzo[d][1,2,3]triazol-1-yl)-1-oxo-1-p-tolylpropan-2-yl nicotinate (A3)
Purified by recrystallization from ethanol; the single crystal suitable for X-ray
measurements was obtained by slow evaporation of ethyl acetate and petroleum
ether (1:1, v/v) at room temperature over 1 week. Yield, 33.5%; yellow solid; m.p.,

Med Chem Res (2010) 19:970–983
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139.5–141.2°C. Anal. Calcd. for C22H18N4O3 (%): C 68.38, H 4.70, N 14.50. Found
(%): C 68.62, H 4.66, N 14.38. mmax/cm-1 (KBr): 3060 (=C–H), 1729, 1690
(s, C=O), 1605 (s, C=N), 1281, 1117 (s, C–O). 1H NMR (CDCl3): 2.45(3H, methyl),
5.53 (2H, CH2), 6.86 (1H, CH), 7.41–9.04 (12H, phenyl H).
3-(1H-Benzo[d][1,2,3]triazol-1-yl)-1-(4-chlorophenyl)-1-oxopropan-2-yl nicotinate
(A4)
Purified by column chromatography using petroleum ether/ethyl acetate (3:1, v/v) as
the eluent. Yield, 44.3%; yellow solid; m.p., 137.8–138.5°C. Anal. Calcd. for
C21H15ClN4O3 (%): C 62.00, H 3.72, N 13.77. Found (%): C 61.89, H 3.88, N 13.76.
m
max/cm-1 (KBr): 3072 (=C–H), 1730, 1692 (s, C=O), 1588 (s, C=N), 1260, 1098 (s,
C–O). 1H NMR (CDCl3): 5.56 (2H, CH2), 6.87 (1H, CH), 7.41–8.20 (12H, phenyl H).
3-(1H-Benzo[d][1,2,3]triazol-1-yl)-1-(2,4-dichlorophenyl)-1-oxopropan-2-yl
nicotinate (A5)
Purified by recrystallization from H2O/acetone (3:1, v/v). Yield, 36.4%; yellow
solid; m.p., 92.8–94.3°C. Anal. Calcd. for C21H14Cl2N4O3 (%): C 57.16, H 3.20, N
12.70. Found (%): C 57.23, H 3.41, N 12.89. mmax/cm-1 (KBr): 3081 (=C–H), 1731,
1
1714 (s, C=O), 1582 (s, C=N), 1113, 1287 (s, C–O). H NMR (CDCl3): 5.29 (2H,
CH2), 6.53 (1H, CH), 7.27–8.74 (11H, phenyl H).
3-(1H-Benzo[d][1,2,3]triazol-1-yl)-1-(2,5-dichlorophenyl)-1-oxopropan-2-yl
nicotinate (A6)
Purified by column chromatography using petroleum ether/ethyl acetate (3:1, v/v) as
the eluent. Yield, 38.5%; yellow solid; m.p., 165.4–166.5°C. Anal. Calcd. for
C21H14Cl2N4O3 (%): C 57.16, H 3.20, N 12.70. Found (%): C 57.52, H 3.17, N
13.04., mmax/cm-1 (KBr): 3089 (=C–H), 1730, 1635 (s, C=O), 1591 (s, C=N), 1272,
1104 (s, C–O). 1H NMR (CDCl3): 5.74 (2H, CH2), 6.88 (1H, CH), 7.29–9.03 (11H,
phenyl H).
3-(1H-Benzo[d][1,2,3]triazol-1-yl)-1-oxo-1-phenylpropan-2-yl isonicotinate (B1)
Purified by column chromatography using petroleum ether/ethyl acetate (3:1, v/v) as
the eluent. Yield, 40.2%; yellow solid; m.p., 166.2–167.5°C. Anal. Calcd. for
C21H16N4O3 (%): C 67.73, H 4.33, N 15.05, Found (%): C 67.85, H 4.37, N 15.67,
m
max/cm-1 (KBr): 3063 (=C–H), 1733, 1687 (s, C=O), 1592 (s, C=N), 1283, 1124 (s,
C–O), 1H NMR (CDCl3): 5.17 and 5.33 (2H, CH2), 6.68 (1H, CH), 7.26–8.73 (13H,
phenyl H).
3-(1H-Benzo[d][1,2,3]triazol-1-yl)-1-oxo-1-p-tolylpropan-2-yl isonicotinate (B3)
Purified by recrystallization from petroleum ether/acetone (2:1, v/v); single crystals
suitable for X-ray measurements were obtained by slow evaporation of acetone and

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Med Chem Res (2010) 19:970–983
alcohol (1:2, v/v) at room temperature over a period of 5 days. Yield, 39.5%; yellow
solid; m.p., 174.2–175.0°C. Anal. Calcd. for C22H18N4O3 (%): C 68.38, H 4.70, N
14.50. Found (%): C 68.71, H 4.58, N 14.82., mmax/cm-1 (KBr): 3053 (=C–H), 1737,
1
1684 (s, C=O), 1605 (s, C=N), 1286, 1125 (s, C–O)., H NMR (CDCl3): 2.46 (3H,
methyl, CH3), 5.38 (2H, CH2), 6.86 (1H, CH), 7.44–8.78 (12H, phenyl H).
3-(1H-Benzo[d][1,2,3]triazol-1-yl)-1-(4-chlorophenyl)-1-oxopropan-2-yl
isonicotinate (B4)
Purified by column chromatography using petroleum ether/ethyl acetate (3:1, v/v) as
the eluent. Yield, 44.8%; yellow solid; m.p., 151.2–152.5°C. Anal. Calcd. for
C21H15ClN4O3 (%): C 62.00, H 3.72, N 13.77. Found (%): C 62.13, H 4.02, N
13.96., mmax/cm-1 (KBr): 3045 (=C–H), 1739, 1688 (s, C=O), 1586 (s, C=N), 1268,
1124 (s, C–O). 1H NMR (CDCl3): 5.57 (2H, CH2), 6.88 (1H, CH), 7.42–8.78 (12H,
phenyl H). ESI–MS m/z 407.1 (MH?).
3-(1H-Benzo[d][1,2,3]triazol-1-yl)-1-(2,4-dichlorophenyl)-1-oxopropan-2-yl
isonicotinate (B5)
Purified by column chromatography using petroleum ether/ethyl acetate (3:1, v/v) as
the eluent; the single crystal suitable for X-ray measurements was obtained by slow
evaporation of acetone and alcohol (1:1, v/v) at room temperature over a period of
several days. Yield, 43.0%; yellow solid; m.p., 115.0–117.2°C. Anal. Calcd. for
C21H14Cl2N4O3 (%): C 57.16, H 3.20, N 12.70. Found (%): C 57.43, H 3.46, N 13.01.
m
max/cm-1 (KBr): 3087 (=C–H), 1736, 1713 (s, C=O, 1581 (s, C=N), 1287, 1118
(s, C–O)., 1H NMR (CDCl3): 5.57 (2H), 6.88 (1H), 7.42–8.78 (11H, phenyl H).
Crystallographic data and structure determination
A colorless crystal, A2 (0.13 9 0.11 9 0.08), a colorless crystal, B3 (0.37 9
0.18 9 0.07), and a colorless crystal, B5 (0.33 9 0.25 9 0.11), were mounted on a
Bruker SMART 1000 CCD area diffractometer for data collection using x-scan
mode. A total of 33,084, 5391, and 11,343 reflections were collected in the range of
1.43 B h B 26.05, 2.08 B h B 26.01, and 1.81 B h B 26.08 using a MoKa
˚
radiation device (k = 0.71073 A) equipped with a graphite monochromator at
293(2) K (Siemens, 1996), of which 11,926, 3668, and 4066 were independent
(Rint = 0.070) and 5006, 2432, and 3235 were observed at I [ 2r(I). Empirical
absorption corrections were carried out using the SADABS (Sheldrick, 1996)
program.
The structures of A2, B3, and B5 were solved by direct methods and refined by
full-matrix least-squares on F2 with the SHELXTL (Sheldrick, 1997) software
package. All non-H atoms were anisotropically refined. All H atoms were located on
difference Fourier maps and refined isotropically. The water hydrogen atoms of B5
were located by means of difference synthesis and refined isotropically. For A2 the
final R = 0.1227 and xR = 0.2897 (x = 1/[r2(Fo2) ? (0.1000P)2 ? 0.0000P],
where P = (F2o ? 2Fc2)/3) with I [ 2r(I). S = 1.441, (D/r)max = 0.001,

Med Chem Res (2010) 19:970–983
975
3
˚
(Dq)max = 0.793, and (Dq)min = -0.630 e/A . For B3 the final R = 0.0568 and
xR = 0.1201 (x = 1/[r2(Fo2) ? (0.0532P)2 ? 0.1631P], where P = (Fo2 ? 2F2c)/3)
with I [ 2r(I). S = 1.035, (D/r)max = 0.001, (Dq)max = 0.186, and (Dq)min = -
3
˚
0.197 e/A . For B5 the final R = 0.0390 and xR = 0.0962 (x = 1/
[r2(F2o) ? (0.0530P)2 ? 0.3967P], where P = (Fo2 ? 2F2c)/3) with I [ 2r(I).
3
˚
S = 1.026, (D/r)max = 0.001, (Dq)max = 0.225, and (Dq)min = -0.220 e/A .
Atomic scattering factors and anomalous dispersion corrections were taken from
International Tables for X-Ray Crystallography (Wilson, 1992). Software packages
used to prepare material for publication were SHELXTL, PARST (Nardelli, 1995),
and PLATON (Spek, 1990).
Biological activities
Testing of primary biological activities was performed in an isolated culture
(Zhang et al., 2007b). Under sterile conditions, 1 ml of sample was added to
culture plates, followed by 9 ml of culture medium. The final concentration was
50 lg/ml. The blank assay was performed with 1 ml of sterile water. A circle of
mycelium with a diameter of 4 mm was cut using a drill. Culture plates were
cultivated at 24 1°C. The extended diameters of the circle mycelium were
measured after 72 h. The relative inhibition rate of the circle mycelium was
compared with a blank assay.
Short-term bioassay of antitumor activity
Cell viability was measured by the MTT (3-[4,5-dimethyl-2-yl]-2,5-diphenyltet-
razolium bromide) method (Hansen et al., 1989). Cells in DMEM were seeded at a
density of 9 9 103 cells per well in 96-well plates for 24 h. After exposure to
different concentrations of compounds dissolved in fresh DMEM for various times,
20 ll of MTT (50 mg/ml) dissolved in DMEM was added. Cells were incubated at
37°C in the dark for 4 h, MTT was removed, and 100 ll of a lysing buffer (10%, w/
v, sodium dodecyl sulfate [SDS] dissolved in a solution of 50% each N,N-dimethyl
formamide and deionized water, pH 4.7) was added. Absorbance at 570 nm was
determined using a microplate reader (SpectraMax190; Molecular Devices) after
shaking in the dark for 15 min. Cell viability was expressed as a percentage of the
cell survival rate.
Results and discussion
Synthesis
The synthesis of compounds A1–A6, B1, and B3–B5 is illustrated in Scheme 1.
Benzotriazolylpropiophenones (II) were prepared according to reported methods
(Wan et al., 2006; Abonia, et al., 2004). Bromine-substituted benzotriazolylpro-
piophenones were obtained in the presence of sodium acetate and acetic acid instead
of AlCl3 (Kasiotis and Haroutounian, 2006); when the ratio of II and sodium acetate

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Med Chem Res (2010) 19:970–983
was 1:2, the reaction temperature was decreased to 30°C to obtain the monobr-
omide. Using triethylamine as the binding acid reagent, nicotinates A1–A6 and
isonicotinates B1 and B3–B5 were obtained at room temperature in good yields
(Scheme 1).
Compounds A1–A6, B1, and B3–B5 were characterized by elemental, IR, and 1H
NMR spectroscopic analysis. Single crystals suitable for X-ray measurements of
compounds A2, B3, and B5 were obtained by slow evaporation of petroleum ether
and alcohol (1:2, v/v), acetone and alcohol (1:2, v/v), and acetone and alcohol (1:1,
v/v), respectively, at room temperature over a period of days.
Crystal structure of compounds A2, B3, and B5
Crystal data and structure refinement for compounds A2, B3, and B5 are listed in
Table 1. Figures 1, 3, and 5 show the molecular structure of A2, B3, and B5, and
their packing diagrams are shown in Figs. 2, 4, and 6, respectively.
In the crystal structure of A2, three independent molecules exist in each unit cell.
˚
The bond lengths of C–N in A2 are in the range of 1.290–1.400 A, shorter than the
˚
˚
single-bond length of 1.48 A and longer than the typical C=N distance of 1.28 A,
indicating a partial double character. This can be interpreted in terms of conjugation
in the benzotriazole ring system. The benzotriazole moieties are nearly coplanar,
O
O
(CH2O)n
N(CH3)2 HCl
R1
R1
HN(CH3)2 HCl
I
N
N
NH
O
O
Br2
N
N
R1
N
N
N
Br
R1
N
O
III
HO
O
HO
II
N
R1=
1, H
N
O
O
2, 4-OCH3
3, 4-CH3
4, 4-Cl
5, 2,4-dichloro
6, 2,5-dichloro
N
O
N
N
O
R1
N
N
R1
O
N
N
O
A
B
N
Scheme 1 Synthesis of title compounds

Med Chem Res (2010) 19:970–983
977
Table 1 Crystal data and structure refinement for A2, B3, and B5
A2
B3
B5
Chem. formula
Fw
C22H18N4O4
402.40
C22H18N4O3
386.40
C21H14Cl2N4O3ÁH2O
459.28
Crystal system
Space group
Monoclinic
P21/c
Triclinic
P-1
Monoclinic
P21/c
˚
a (A)
23.059(4)
9.3484(17)
29.986(5)
90
9.840(2)
10.050(2)
10.994(2)
114.635(3)
97.113(4)
98.384(4)
956.9(3)
2
11.2644(11)
12.8260(13)
14.2814(14)
90
˚
b (A)
˚
c (A)
a (°)
b (°)
c (°)
110.433(3)
90
92.659(2)
90
3
˚
V (A )
6057.2(18)
12
2061.1(4)
4
Z
q (g/cm3)
1.324
1.341
1.480
l (mm-1
)
0.116
0.092
0.352
F (000)
2520
404
944
Abs. correct
SABABS
Colorless
0.13 9 0.11 9 0.08
1.43–26.05
33,084
SABABS
Colorless
0.37 9 0.18 9 0.07
2.08–26.01
5,391
SABABS
Colorless
0.33 9 0.25 9 0.11
1.81–26.08
11,343
Cryst. color
Cryst.l size (mm3)
h
No. reflects meads
No. unique reflects
11,926
3,668
4,066
No. params
361
262
288
R
0.1227
0.0568
0.0390
xR
S
0.3327
0.1394
0.1039
1.44
1.03
1.03
Fig. 1 Molecular structure of A2

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Med Chem Res (2010) 19:970–983
Fig. 2 Packing diagram of A2 along the c axis
Fig. 3 Molecular structure of
B3
Fig. 4 Packing diagram of B3 along the b axis
with dihedral angles of 0.8(3)°, 1.8(3)°, and 2.1(3)° between the phenyl plane and its
fused triazole plane. The C7–C8–C9, C29–C30–C31, and C51–C52–C53 angles are
109.5(1)°, 109.6(1)°, and 111.2(1)°, respectively, determined by the sp3

Med Chem Res (2010) 19:970–983
979
Fig. 5 Molecular structure of
B5
Fig. 6 Packing diagram of B5 along the c axis
hybridization state of C8, C30, and C52, respectively. In a unit cell, three
independent molecules are connected by C–HÁÁÁO and C–HÁÁÁN hydrogen bonding
interactions.
For B3, all bond lengths and angles are within normal ranges. The angle between
the C10–C15 phenyl plane and its fused triazole plane is 0.86(1)°, which indicates
that the two planes are essentially coplanar, but the benzotriazole ring system makes
a dihedral angle of 87.41(1)° and 18.63(1)° with the pyridine ring and the other
phenyl plane, respectively, while the dihedral angle between the latter aromatic

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Med Chem Res (2010) 19:970–983
rings is 88.84(1)°. As a result, the whole compound is not a planar molecule. In the
crystal structure, two molecules are linked by two mutual intermolecular hydrogen
bonds, C15–H15AÁÁÁO1, resulting in a centrosymmetric dimer. Furthermore, the
dimers are linked by another intermolecular hydrogen bond, C21–H21AÁÁÁN2,
leading to the three-dimensional network structure.
In the structure of B5, the asymmetric unit consists of a monomeric compound
and one water molecule, which are incorporated during recystallization. The bond
lengths and angles show normal values, which are comparable to those in A2 and
B3. Within the asymmetric unit, the water acts as both hydrogen bond donor and
hydrogen bond acceptor. In the crystal structure, molecules are linked into dimers
by intermolecular hydrogen bonds, C14–H4BÁÁÁO3. Furthermore, the crystal
packing is stabilized by p–p stacking interactions with a distance of Cg1ÁÁÁCg2
˚
(1–x, -1/2 ? y, 1/2–z) of 3.671 A, where Cg1 and Cg2 denote the centroids of rings
N1–N3/C10/C11 and N4/C17–C21, respectively.
Biological activities
The biological activities of compounds A1–A5 and B1 were tested, including their
fungicidal inhibiting activities at 50 lg/ml. Experimental results on the fungicidal
inhibiting activities of these compounds are reported in Table 2. On the whole, they
exhibit less efficiency on tomato early blight, peanut Cercosporium arachidicola,
and asparagus stem wilt and better efficiency on apple ringspot.
Antitumor biological activities
Antitumor biological activities of compounds were also tested by the method of
MTT. Experimental results show that compounds A3 and A4 have antitumor
activity for hepatoma 7402 cells and breast cancer 4T-1 cells. Results of short-term
bioassay of antitumor activity of compounds A3 and A4 are presented in Figs. 7 and
8. They show that compound A3 affects liver cancer 7402 cell viability, and the
liver cancer 7402 cell was inhibited. The results also show that the toxicity of
compound A3 has a time-quantity efficiency relationship (Fig. 7).
MTT assay showed that application of A4 induced dose-dependent cell
damage (Fig. 8). While A4 at a concentration of 0.2 mM was used to estimate a
Table 2 Fungicidal activities of compounds A1, A3–A5, and B1
Compound
Fungicidal activity (c = 0.005% inhibition)
Tomato early
blight
Peanut Cercosporium
arachidicola
Apple
ringspot
Stem wilt
of asparagus
A1
A3
A4
A5
B1
8.3
10.0
15.0
35.0
7.4
15.0
0
56.8
24.4
19.5
31.7
36.7
14.3
21.4
28.6
21.4
0
18.8
31.3
13.0

Med Chem Res (2010) 19:970–983
981
Fig. 7 Effects of different
concentrations of compound A3
on survival rates of hepatoma
7402 cells
70
65
60
55
50
45
40
35
24 h
48 h
72 h
0
50
100
150
200
Concentration (10-6M)
Fig. 8 Effects of different
concentrations of compound A4
on survival rates of breast cancer
4T-1 cells
80
75
70
65
60
55
50
45
40
24 h
48 h
72 h
0
20 40 60 80 100 120 140 160 180 200 220
Concentration (10-6M)
time-dependent viability loss, cell viability after incubation with A4 at concentra-
tions of 0.2 mM for 24, 48, and 72 h of treatment was 52.2, 45.0, and 40.3% of the
control value, respectively. Results of the MTT assay showed that A4 treatment
significantly reduced the viability of the breast cancer cell 4T-1, and the toxicity of
compound A4 has a time-quantity efficiency relationship (Fig. 8).
Studies on the mechanism of antitumor activity are ongoing.
Acknowledgments This work was supported by the Natural Science Foundation of Shandong Province
(Grants Y2007B50 and Y2006B10), Outstanding Adult-Young Scientific Research Encouraging
Foundation of Shandong Province (2008BS09017), and Qingdao University of Science and Technology
Dr. Seed funding.
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