Pyran Derivatives: Anti-Breast Cancer Activity and Docking Study

Article abstract of DOI:10.1134/S1070363218120307

Four novel pyran derivatives 1–4 are synthesized and characterized by IR, ¹H NMR, HRMS, and single crystal X-ray data. Anticancer activity of the compounds is tested against four human breast cancer cells (HCC1937, HCC70, MDA-MB-436, and BT474)

Full text of DOI:10.1134/S1070363218120307

ISSN 1070-3632, Russian Journal of General Chemistry, 2018, Vol. 88, No. 12, pp. 2664–2668. © Pleiades Publishing, Ltd., 2018.
Pyran Derivatives: Anti-Breast Cancer Activity
and Docking Study1
H. Hana, Z.-F. Zhangb, J.-F. Zhangc, and B. Zhangc*
a Inner Mongolia University For The Nationalities, Tongliao, Inner Mongolia Autonomous Region, China
b Nursing Department, Tongliao City Hospital, Tongliao, Inner Mongolia Autonomous Region, China
c The First Affiliated Hospital of Inner Mongolia University For The Nationalities,
Tongliao, Inner Mongolia Autonomous Region, China
*e-mail: bin_zhang666@126.com
Received October 23, 2018
Revised December 10, 2018
Accepted December 10, 2018
1
Abstract—Four novel pyran derivatives 1–4 are synthesized and characterized by IR, H NMR, HRMS, and
single crystal X-ray data. Anticancer activity of the compounds is tested against four human breast cancer cells
(HCC1937, HCC70, MDA-MB-436, and BT474) by MTT assay. Molecular docking study supports the
biological assay data and suggests that compound 2 has a potential as an anticancer agent.
Keywords: pyran, anticancer activity, molecular docking study
DOI: 10.1134/S1070363218120307
Dihydropyrans represent an active class of com-
pounds demonstrating a wide range of biological
activities, including anticoagulant, insecticidal, anthel-
minthic, hypnotic, antifungal, and HIV protease
inhibiting [1, 2]. Such activities made dihydropyrans to
be attractive targets of organic synthesis [3]. In the
current study four new pyran derivatives 1–4 (Fig. 1)
were synthesized and their potential anti-tumor activity
tested.
refluxed for 2–3 h, then cooled down to room
temperature. The precipitated products were filtered
off and washed with ice-cold water and ethanol, and
then dried under vacuum.
2-Amino-4-(4-methylphenyl)-5-oxo-5,6,7,8-tetra-
hydro-4H-chromene-3-carbonitrile (1). mp 221–
222°C. IR spectrum, ν, cm–1: 3021, 2634, 2421, 1712,
1601, 985. 1H NMR spectrum, δ, ppm: 6.989–7.073 m
(6H), 4.138 s (1H), 2.585–2.623 m (2H), 2.250–2.305
m (5H), 1.869–1.978 m (2H). HRMS (ESI+): m/z:
calculated for C17H16N2O2: 303.1104 [M + Na]+;
found: 303.1123.
EXPERIMENTAL
IR spectra (KBr discs) were recorded on a Brucker
1
Equinox-55 spectrophotometer. H NMR spectra were
2-Amino-4-(2-nitrophenyl)-5-oxo-5,6,7,8-tetra-
hydro-4H-chromene-3-carbonitrile (2). mp 234–
235°C. IR spectrum, ν, cm–1: 3066, 2711, 2533, 1710,
measured on a Varian Inova-400 spectrometer using
DMSO-d6 as a solvent. Mass spectra were measured on
a micrOTOF-Q II mass spectrometer. Melting points
were determined on a XT-4 micro melting apparatus.
1
1601, 980. H NMR spectrum, δ, ppm: 7.800–7.823 q
(1H), 7.634–7.656 q (1H), 7.374–7.451 m (2H), 7.202
s (2H), 4.933 s (1H), 2.578–2.608 t (2H), 2.133–2.255
m (2H), 1.824–1.953 m (2H). HRMS (ESI+): m/z:
calculated for C16H13N3O4: 334.0798 [M + Na]+;
found: 334.0783.
Synthesis of compounds 1–4. The compounds 1–4
were synthesized according to a developed earlier
procedure [4]. A mixture of 3,5-cyclohexanedione
(10 mmol) with aromatic aldehydes (10 mmol),
malononitrile (10 mmol) and 4-(dimethylamino)
pyridine (DMAP) (1 mmol) in ethanol (100 mL) was
Ethyl 2-amino-4-(4-bromophenyl)-5-oxo-5,6,7,8-
tetrahydro-4H-chromene-3-carboxylate (3). mp 198–
199°C. IR spectrum, ν, cm–1: 3712, 2612, 2319, 1723,
1
1612, 972. H NMR spectrum, δ, ppm: 7.592 s (2H),
1 The text was submitted by the authors in English.
7.386–7.400 q (2H), 7.106–7.120 d (2H), 4.505 s (1H),
2664

PYRAN DERIVATIVES: ANTI-BREAST CANCER ACTIVITY
NH2
2665
NH2
NH2
NH2
COOEt
CN
NO2
COOEt
CN
O
O
O
O
O
Br
O
O
F
O
1
2
3
4
Fig. 1. Formulas of compounds 1–4.
3.945–3.959 q (2H), 2.602–2.619 t (2H), 2.276–2.331
m (1H), 2.200–2.245 m (1H), 1.932–1.972 m (1H),
1.810–1.842 m (1H), 1.070–1.093 t (3H). HRMS
(ESI+): m/z: calculated for C18H18BrNO4: 414.0317
[M + Na]+; found: 414.0321.
Ethyl 2-amino-4-(4-fluorophenyl)-5-oxo-5,6,7,8-
tetrahydro-4H-chromene-3-carboxylate (4). mp 200–
201°C. IR spectrum, ν, cm–1: 3371, 2712, 2541, 1821,
1611, 921. 1H NMR (DMSO-d6, δ, ppm): 7.568 s (2H),
7.161–7.185 m (2H), 7.000–7.030 t (2H), 4.535 s (1H),
Table 1. Crystal data and structure refinement for compounds 1 and 2
Value
Parameter
1
2
C16H13N3O4
311.29
Formula
Mr
C17H16N2O2
280.32
Temperature, K
Crystal system
Space group
a, Å
293(2)
293(2)
Triclinic
P-1
Triclinic
P-1
8.5783 (8)
8.7427 (8)
11.0298 (9)
72.652 (8)
69.856 (8)
80.054 (8)
739.01 (11)
2
8.8933 (6)
9.3992 (9)
10.9991 (9)
73.190 (8)
85.811 (6)
78.133 (7)
861.23 (12)
2
b, Å
c, Å
α, deg
β, deg
γ, deg
V, Å3
Z
D
calc, g/cm3
1.260
1.200
μ(MoKα), mm–1
0.084
0.088
θ range, deg
2.45–24.99
4709
2.56–25.00
5674
Reflections collected
Number of unique data (Rint)
Number of data with I ≥ 2σ(I)
R1
2600 (0.0221)
1889
3032 (0.0291)
2169
0.0506
0.0619
ωR2 (all data)
0.1714
0.2028
CCDC
1559701
1559700
RUSSIAN JOURNAL OF GENERAL CHEMISTRY Vol. 88 No. 12 2018

2666
HAN et al.
Table 2. Hydrogen-bond geometry (Å) for compounds 1 and 2
Comp. no.
D–H···A
N1–H1A···N2
N1–H1B···O2
C12–H12A···N2
N1–H1A···N2
C3–H3A···O3
C3–H3A···N3
D–H
H···A
D···A
Symmetry code
1 – x, 1 – y, 1 – z
–1 + x, y, z
∠D–H···A, deg
176.00
1
0.8600 2.3100 3.167(3)
0.8600
0.9300
2.1600 2.997(2)
2.5100 3.258(3)
164.00
138.00
2 – x, 1 – y, 1 – z
–x, 1 – y, 2 – z
–x, 1 – y, 2 – z
–x, 1 – y, 2 – z
2
0.8600 2.2000 3.057(3)
172.00
0.9800
0.9800
2.2000 2.920(3)
2.5900 3.041(3)
129.00
108.00
3.950–3.962 d (2H), 2.596–2.624 m (2H), 2.214–2.305
m (2H), 1.935–1.966 m (1H), 1.808–1.848 m (1H),
1.066–1.090 t (3H). HRMS (ESI+): m/z: calculated for
C18H18FNO4: 354.1118 [M + Na]+; found: 354.1129.
culture plates and incubated for 24 h at 37°C in a CO2
incubator. Compounds were deliquated to the part-
icular concentrations in culture medium and added to
the wells with respective vehicle control. After 48 h of
incubation, MTT (10 μL, 5 mg/mL) was replenished
into each well and the plates continued to be incubated
for 4 h more. Then the supernatant from each well was
carefully removed, compounds were dissolved in
DMSO (100 μL) and absorbance at 570 nm was
recorded.
Crystal structure. Suitable single crystals of
compounds 1 and 2 were crystallized from chloroform
(Table 1). Diffraction data were acquired on a Bruker
Smart Apex CCD area detector using a graphite
monochromated MoKα radiation (λ = 0.71073 Å) at
room temperature. The structure was solved by using
the program SHELXL-97 [5]. All hydrogen atoms
were added academically.
Simulation details. Autodock Vina v1.2 was used
in the study of the binding mode of compounds 1 and 2
with tubulin [7]. The geometry structures of the
compounds were optimized by quantum chemistry
calculations by Gaussian 09, B3LYP theory (6-31g*
basic set). AutoDockTools v1.5.6 were used to transfer
1AS0 and optimize the structures. Only polar
hydrogens of the structures were considered. The
center coordinates of search grid of tubulin were set to
MTT assay. The 3-(4,5-dimethylthiazol-2-yl)-2,5-
diphenyltetrazolium bromide (MTT) assay [6] was
used in determination of anticancer activity of
compounds 1 and 2 and doxorubicin. First 1×104 cells
per well were seeded in Dulbecco's modified Eagle's
medium (DMEM, 100 μL) supplemented with 10%
fetal bovine serum in each well of 96-well micro-
(a)
(b)
Fig. 2. Molecular structures of compounds (a) 1 and (b) 2.
RUSSIAN JOURNAL OF GENERAL CHEMISTRY Vol. 88 No. 12 2018

PYRAN DERIVATIVES: ANTI-BREAST CANCER ACTIVITY
(a) (b)
2667
Fig. 3. The crystal packing structure of (a) compound 1 along the a axis and (b) compound 2 along the c axis.
38.15, –26.99, and 6.18, length of the search grid was
15. All parameters needed by autodock vina were used
as default if not mentioned specifically. The calculated
data were analyzed and visualized by PyMoL v1.8.6.
distances were in their normal ranges and could be
comparable with the reported compounds.
In vitro anticancer screening. Evaluation of
compounds 1–4 for their in vitro cytotoxic activity
against four human breast cancer cells (HCC1937,
HCC70, MDA-MB-436, and BT474) was carried out
using doxorubicin as the reference drug. Cytotoxicity
was assessed at concentrations of 0, 0.01, 0.1, 10 and
100 μg/mL. The relation between remaining fraction
and drug concentration was plotted to obtain survival
curves of the three tumor cell lines after accretion of
the specified compounds.
RESULTS AND DISCUSSION
Crystal structures of compounds 1 and 2 were
studied by X-ray diffraction single-crystal structural
analysis. Due to similarity of two structures, the
structure of compound 1 was selected as the sample in
this study. The method demonstrated that the
compound 1 belonged to the triclinic crystal system
with the space group of P-1. The compound 1 two-
dimensional supra-molecular structure was determined
by three types of H-bond: N–H···N, N–H···O and
C–H···N (Table 2). The molecular structural units of
compounds 1 and 2 are presented in Fig. 2 and their
crystals packing in Fig. 3. The estimated bond
Comparison of IC50 values determined for the
compounds 1–4 and the reference drug are listed in
Table 3. According to the accumulated data,
compound 2 demonstrated the highest cytotoxic effect
against the four human cancer cells, and it was close to
that of the reference drug.
Table 3. Results of in vitro cytotoxic activity of 1–4 and doxorubicin on four human breast cancer cells
IC50, μM
Compound
HCC1937
HCC70
25
MDA-MB-436
BT474
25
1
30
15
25
10
2
10
8
3
>100
>100
8
>100
>100
8
>100
>100
10
>100
>100
10
4
Doxorubicin
RUSSIAN JOURNAL OF GENERAL CHEMISTRY Vol. 88 No. 12 2018

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HAN et al.
(a)
(b)
Fig. 4. The interaction mode of compounds (a) 1 and (b) 2 with tubulin at the colchicine binding site.
Molecular docking. The possible binding modes of
CONFLICT OF INTEREST
No conflict of interest was declared by the authors.
REFERENCES
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by autodocking. As shown in Fig. 4, the fused pyran
and cyclohexanone rings of compounds 1 and 2
interacted with the surrounding protein molecule. Two
hydrogen bonds were formed by oxygen atom of
carbonyl group and hydrogen atom of the amino
groups of proteins (GLU-276 and LYS-277). For
compound 1, the bond length values were determined
to be 2.4 and 2.0 nm respectively; for compound 2, the
lengths of both H-bonds were 2.3 nm. The hydrogen
bonds between compound 2 and the protein was
determined to be stronger than that of compound 1,
which could be supported by the lowest free binding
energy values for compounds 1 and 2 (–7.5 and
–8.8 kcal/mol respectively).
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CONCLUSIONS
Novel pyran derivatives 1–4 are synthesized and
their anti-tumor activity is studied. The compound 2
demonstrates the most potent anti-cancer activity,
supported by molecular docking simulation which
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ACKNOWLEDGMENTS
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Natural Science Foundation of China (no. 81373573).
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