Novel bis(dihydropyrano[3,2-c]chromenes): Synthesis, Antiproliferative Effect and Molecular Docking Simulation

Article abstract of DOI:10.1002/jhet.3072

An efficient and convenient route for the synthesis of novel bis dihydropyrano[3,2-c]chromenes is reported. The synthetic pathway involves one-pot, multicomponent reaction of bis-aldehydes, malononitrile, and 4-hydroxycoumarin in the presence of pyridine

Full text of DOI:10.1002/jhet.3072

Month 2017
Novel bis(dihydropyrano[3,2-c]chromenes): Synthesis, Antiproliferative
Effect and Molecular Docking Simulation
Amna M. Abdella, Magda F. Mohamed, Aly F. Mohamed, Ahmed H. M. Elwahy, * and Ismail A. Abdelhamida*
a
b
c
a
a
Chemistry Department, Faculty of Science, Cairo University, Giza, Egypt
b
Department of Chemistry (Biochemistry Branch), Faculty of Science, Cairo University, Giza, Egypt
c
Company for Production of Vaccines, Sera and Drugs (VACSERA), Giza, Egypt
*
E-mail: aelwahy@hotmail.com; ismail_shafy@yahoo.com
Received August 9, 2017
DOI 10.1002/jhet.3072
Published online 00 Month 2017 in Wiley Online Library (wileyonlinelibrary.com).
An efficient and convenient route for the synthesis of novel bis dihydropyrano[3,2-c]chromenes is re-
ported. The synthetic pathway involves one-pot, multicomponent reaction of bis-aldehydes, malononitrile,
and 4-hydroxycoumarin in the presence of pyridine or acetic acid/sodium acetate. A stepwise approach for
the synthesis of the target compounds was also investigated. The anticancer activity of the synthesized prod-
ucts against MCF7, HEPG2, and A549 cell lines was assessed. Attempts to detect the molecular action of 6g,
docking simulation was done using DHFR PDB:ID (1DLS). The study revealed that compound 6g was
strongly fit into the active sites of the target protein through six bindings, and hence, it was considered as
promising inhibitor for cancer proliferation.
J. Heterocyclic Chem., 00, 00 (2017).
INTRODUCTION
of derivatized heterocycles, the most active library
compounds had a bis-heterocyclic structure [21–24].
In continuation of these studies, we report on the first
Dihydropyrano[3,2-c]chromenes are very interesting
compounds due to their biological and pharmaceutical
activities, for instance, as anti-HIV, antimicrobial,
antibacterial, anticancer, spasmolytic, and anticoagulant
synthesis
of
novel
bis(2-amino-5-oxo-4,5-
dihydropyrano[3,2-c]chromene-3-carbonitrile) derivatives
in which the two dihydropyrano[3,2-c]chromene moieties
are linked to aliphatic or aromatic spacer. The anticancer
activity of the novel synthesized products against MCF7,
HEPG2, and A549 cell lines was also investigated.
[
1–3]. Dihydropyrano[3,2-c]chromene derivatives have
likewise been accounted for as potential medications for
the treatment of neurodegenerative diseases, including
Alzheimer’s sickness, Parkinson’s ailment, amyotrophic
lateral sclerosis, AIDS-associated dementia, Down’s
syndrome, and Huntington’s disease and in addition,
schizophrenia and myoclonus [1–3].
RESULTS AND DISCUSSION
Moreover, multicomponent reactions, which include
more than two components are very elegant and fastest
route for the synthesis of complex molecules. They
reduce the number of reaction steps, minimize by-
products formation, and thus result in both atom and step
economy [4–10]. Recently, we reviewed the use of
multicomponent reactions in the synthesis of fused
heterocyclic systems [11–14].
Besides, bis-heterocyclic compounds were found to
have various applications as electrical materials [15],
chelating agents, and metal ligands [16]. Attention has
also progressively been paid lately to the synthesis of
Synthetic chemistry. Previously, impressive endeavors
have been made in the synthesis of pyrano[3,2-c]
chromenes 4 as an important class of compounds. Several
methods have been accounted for to the synthesis of this
class of compounds, including the reaction of aromatic
aldehydes 1, malononitrile 2, and 4-hydroxycoumarin 3
in the presence of homogeneous or heterogeneous
catalysts under conventional heating, microwave
irradiation, or ultrasound irradiation (Scheme 1) [25–37].
As part of the continuing interest in this area, we
investigated here two possible synthetic approaches, path
A and path B for bipodal dihydropyrano[3,2-c]chromenes
6, as outlined in Scheme 2. If there should arise an
occurrence of pathway A, the desired dihydropyrano[3,2-
c]chromene 4 has initially been synthesized, which may
then be reacted with the appropriate dihalo compounds 5
to give the desired bipodals by using a mild base. On the
heterocyclic
compounds,
especially
when
two
pharmacologically active heterocyclic moieties are
present in the same molecule [17–19]. These compounds
were encountered in numerous bioactive natural product
[20], and recent reports demonstrated that among libraries
©
2017 Wiley Periodicals, Inc.

A. M. Abdella, M. F. Mohamed, A. F. Mohamed, A. H. M. Elwahy, and I. A. Abdelhamid
Vol 000
other hand, this can likewise be accomplished by means of
path B, where bis-aldehydes 7 have to be synthesized in a
first step, which can further be reacted with malononitrile,
and 4-hydroxycoumarin to give the desired bipodals 6
unsuccessful, and instead, 3-cyanocoumarin 8 was obtained
in 74% yield as a sole product [39,40].
Searching for an expedient pathway to prepare the target
compounds 6, we turned to path B, and the bis(aldehydes)
7 were chosen as key intermediates. The latter compounds
should then react effectively with two equivalents of both
malononitrile 2 and 4-hydroxycoumarin 3 under basic
conditions to give the bis(4H-chromene-3-carbonitrile) 6.
Our preliminary investigations were focused on
systematic evaluation of different catalysts for the model
(Scheme 2).
As indicated by the first methodology, the precursor
monopodal dihydropyrano[3,2-c]chromene 4a was
prepared via initial Knoevenagel condensation of the
appropriate 4-hydroxybenzaldehyde 1a with malononitrile
in ethanol in the presence of piperidine as a catalyst to
give the corresponding arylidenemalononitrile derivative
followed by reaction with 4-hydroxycoumarin 3 via
Michael addition reaction [36,38]. The potassium salt
0
reaction of 2,2 -(ethane-1,2-diylbis(oxy))dibenzaldehyde
7a with two equivalents of both malononitrile 2 and 4-
hydroxycoumarin 3 (Scheme 4 and Table 1). At first, we
attempted to utilize chitosan in dioxane as an ecofriendly
basic catalyst for the multicomponent reaction.
Unfortunately, the reaction stopped at the ylidene stage 9,
and no traces of the of target bis(4H-chromene-3-
carbonitrile) 6 were detected even after prolonged
heating. Similar results were obtained when we explored
the catalytic activity of phthalimide-N-oxyl anion as an
effective and readily available Lewis base for the above
reaction. When the reaction was carried out in refluxing
dioxane for 6 h using 1,4-diazabicyclo[2.2.2]octan
(DABCO) as an inexpensive, ecofriendly, and nontoxic
(obtained upon treatment of 4a with ethanolic KOH) was
then allowed to react with the appropriate dihalo
compound 5 in refluxing DMF. The reaction, unfortunately,
did not lead to the formation of the corresponding bis(4H-
chromene-3-carbonitrile) 6 and gave instead a mixture of
products that were not easily separated and have not been
characterized yet (Scheme 3). It is noteworthy to mention
that our attempts to synthesize 2-amino-4-(2-
hydroxyphenyl)-5-oxo-4,5-dihydropyrano[3,2-c]chromene-
3
-carbonitrile 4b via multicomponent reaction of
salicylaldehyde 1b with malononitrile and 4-
hydroxycoumarin under basic conditions were
1
base catalyst, the H–NMR indicated the presence of a
Scheme 1. The synthesis of pyrano[3,2-c]chromenes 4.
Scheme 2. The possible synthetic approaches for bipodal dihydropyrano[3,2-c]chromenes 6.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2017
Synthesis of Novel bis(dihydropyrano[3,2-c]chromenes)
Scheme 3. Trials to synthesize the bis(4H-chromene-3-carbonitrile) 6 through the bis alkylation of the potassium slat of compound 4a with the di-bromo
compounds.
Scheme 4. Attempted synthesis of the bis(4H-chromene-3-carbonitrile) 6 in EtOH / piperidine.
mixture of the corresponding bis-arylidenemalononitriles 9
and the target compound 6 at a ratio of 1:1 based on the
area of their characteristic signals. The product ratio did
not change with further standing up to 12 h. The same
results were obtained using piperidine as a basic catalyst
in either refluxing ethanol or refluxing dioxane. The
reaction was also investigated in the presence of a
catalytic amount of hexamethylenetetramine as a very
cheap, nontoxic, and stable reagent followed the method
recently reported by Wang et al. [31] to prepare a variety
of dihydropyrano[3,2-c]chromene derivatives in high to
excellent yields within short times. We also tried the
ecofriendly methodology recently reported by Sajadikhah
et al. [41] in which NaCl in water–ethanol media (3:1)
was used as an inexpensive catalyst for the one-pot
synthesis of dihydropyrano[3,2-c]chromene derivatives
[41]. Unfortunately, in both cases, no traces of the target
bis(dihydropyrano[3,2-c]chromenes) 6 were obtained. On
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

A. M. Abdella, M. F. Mohamed, A. F. Mohamed, A. H. M. Elwahy, and I. A. Abdelhamid
Vol 000
ꢀ
1
Table 1
group appeared as a broad band at 1671 cm . The
Optimizing the yield of compound 6a.
constitutions of compounds 6a–l were established based
1
on their elemental analysis and spectral data. The
H
Conversion
a
NMR spectrum of 6a indicated the presence of the pyran-
H4 as singlet signal at δ 4.62 ppm. Moreover,
compounds 6 also featured the methylene ether linkage
Entry
Solvent
Catalyst
6:9
(%)
1
2
3
4
5
6
7
Dioxane
Water
Chitosan
PINO
DABCO
Piperidine
Piperidine
HMT
0:100
0:100
50:50
50:50
50:50
0:100
0:100
80
78
85
87
89
82
76
Dioxane
Dioxane
Ethanol
Dioxane
Water–
Ethanol
OCH as multiplet signals in the region 3.50–5.27 ppm,
2
although their precursors 7 or 9 exhibit singlet signals for
these protons. This suggests that the generated
asymmetric center (in the dihydropyran rings) is close
NaCl
enough to this CH group to affect such splitting. All
2
8
9
Pyridine
Acetic acid
100:0
100:0
88
82
other protons were seen at the expected chemical shifts
Sodium
acetate
and
integral
values
(See
Experimental
section and Supporting Information).
a
1
Based on % conversion of 7a, 2, and 3 into products 6a and 9a on H
NMR spectroscopic analysis of crude reaction mixtures.
Biology. Anticancer evaluations.
In the present
investigation, all the synthesized compounds were
screened against the three cell lines MCF7, HEPG2, and
A549 (Fig. 1). The concentration causing 50% cell
growth inhibition (IC ) was determined as shown in
the other hand, the reaction takes another course when
performed in refluxing pyridine for 15 min, and the
bis(4H-dihydrobenzo[b]pyrans) 6a could be obtained in
50
(Table 3). It was shown that the breast line was the most
sensitive one toward most of our derivatives. Also, it was
noted that compound 6g was the most active compound
in this series with IC50 values (0.03, 0.08, and 0.22 mM)
against MCF7, HEPG2, and A549 cell lines, respectively.
On the other hand, compound 6b was the least active one
toward HEPG2 and A549 lines with IC50 (9.99 and
4.9 mM), respectively. Regarding compounds 6c
(IC50 = 1.07, 0.31, and 0.73 mM) and 6d (IC50 = 2.38,
0.77, and 0.78), it was noted that the presence of
unsaturated butene group in 6d decreases the biological
activity as indicated in the IC50 values against A549,
MCF7, and HEPG2, respectively. Unfortunately, the
activity of 6f was decreased to (8.9 mM) using HEPG2
cell line. The data also indicate that compound 6f with
para substitution (IC50 = 0.04 and 0.65 mM) is more
favored than 6e with ortho substitution (IC50 = 0.06 and
4.03 mM) toward the two cell lines MCF7 and A549,
respectively. Moreover, it is clear that the addition of
bromines into the benzene ring increases the activity as
indicated in the IC50 values [6f (IC50 = 0.65, 0.04, and
8.9 mM), 6g (IC50 = 0.22, 0.03, and 0.08 mM)] against
A549, MCF7, and HEPG2, respectively. Compound 6h
with meta substitution showed better cytotoxicity to
MCF7 and HEPG2, cell lines (IC50 = 0.52 and 0.57) than
8
8% yield as a sole product. Compound 6a was
alternatively obtained in 82% yield by carrying out the
reaction in refluxing acetic acid containing a catalytic
amount of sodium acetate for 1 h.
Subsequently, with optimal condition in hand, the
generality and synthetic scope of this reaction were
demonstrated by synthesizing
a series of bis(4H-
chromene-3-carbonitriles) 6a–l (linked to aliphatic or
aromatic cores via ether linkage). Thus, different bis-
aldehydes 7a–f [42–44] were well tolerated under the
optimized reaction conditions and furnished the
corresponding bis(4H-chromene-3-carbonitriles)
good yields (Scheme 5 and Table 2).
It is even possible to carry out these reactions in a
stepwise fashion in which the arylidenemalononitrile
derivatives 9, [45] containing the electron-poor C–C
double bond, are firstly produced by Knoevenagel
condensation of the bis-aldehydes 7 with two moles of
malononitrile. Subsequent reaction of 9 two moles of 4-
hydroxycoumarin 3 afforded the target molecules 6 good
yields (Scheme 6, Table 2, method C).
6
in
The infrared (IR) spectra of compound 6a indicated the
ꢀ
1
presence of amino group at 3402 cm . In addition, it
revealed the cyano band at 2202 cm . The carbonyl
ꢀ
1
Scheme 5. Synthesis of a series of bis(4H-chromene-3-carbonitrile) 6a–i under the optimized reaction conditions.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2017
Synthesis of Novel bis(dihydropyrano[3,2-c]chromenes)
Table 2
The % yields of the bis(4H-chromene-3-carbonitriles) 6 in different reaction conditions.
%
Yield
a
b
c
Compound
X
Y
Method A
Method B
Method C
6
6
6
6
a
b
c
H
H
H
H
-(CH
-(CH
-(CH
2
2
2
)
2
)
3
)
4
-
-
-
88
86
89
86
82
82
84
75
85
84
86
82
d
6e
H
89
81
86
6
6
6
f
H
87
88
87
80
83
80
82
-
g
h
Br
Br
-
6i
H
90
82
-
a
Bisaldehyde (1 mmol) 7, malononitrile 2 (2 mmol), and 3 (2 mmol) in pyridine.
Bisaldehyde (1 mmol) 7, malononitrile 2 (2 mmol), and 3 (2 mmol) in AcOH – AcONa.
b
c
Bis-arylidenemalononitrile (1 mmol) 9, and 3 (2 mmol) in pyridine.
-, position of attachment of the two rings.
Scheme 6. Stepwise synthesis of the bis(4H-chromene-3-carbonitrile) 6.
for A549 (IC50 = 1.19), respectively. Finally, it is clear that
while the naphthalene ring in 6i decreases the biological
activity against the respective MCF7 and HEPG2
present in the crystal structure. The solvent molecules
were deleted, and the bond order of the crystal ligand
and protein was adjusted. The ligand was built using the
chembiodraw ultra 10.0, protonate 3D and subjected to
energy minimization. Docking study of compound 6g
into the active site of human dihydrofolate reductase
enzyme showed six interactions as showed in Figure 2.
The first interaction was hydrogen bonding between the
amino acid lys 63 and N atom of CN group with bond
distance 2.64A°. The second binding was hydrogen
(IC50 = 3.72 and 0.16 mM), it increases the activity in
case of A549 (IC50 = 0.13 mM).
Molecular docking simulation.
Molecular modeling
was performed on human dihydrofolate reductase
enzyme to predict the binding mode of 6g within the
binding site of target proteins (human dihydrofolate
reductase enzyme). Protein was selected and downloaded
from the Protein Data Bank (PDB ID: 1DLS). The
protein was optimized, removing the water molecule and
cofactors from the proteins and deleting the ligand
bonding between ser 59 and H atom of NH group with
2
bond distance 2.03A°. The third and fourth bindings
were H-bonding between Arg 28 and the two oxygen
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

A. M. Abdella, M. F. Mohamed, A. F. Mohamed, A. H. M. Elwahy, and I. A. Abdelhamid
Vol 000
Figure 1. Cytotoxic activity of the new derivatives (Doxorubicin, 6a–i) against the tumor A549, MCF7, and HEPG2 cell lines after 48-h exposure. Doxo-
rubicin is used as a standard agent against the same lines. Each point is the mean ± standard deviation of three independent experiments performed in
triplicate, using prism software program (Graphpad software incorporated, version 3). [Color figure can be viewed at wileyonlinelibrary.com]
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2017
Synthesis of Novel bis(dihydropyrano[3,2-c]chromenes)
Table 3
EXPERIMENTAL AND ANALYTICAL DATA
The IC50 values (the drug concentrations that inhibited 50% of cell pro-
liferation) of the selected nine compounds on the different human cell
Chemistry. General. Melting points were measured
with a Stuart melting point apparatus and are uncorrected.
2
lines A549, MCF7, and HEPG .
IC50 values (mM)
MCF7
The IR spectra were recorded using an FTIR Bruker–
1
vector 22 spectrophotometer as KBr pellets. The
H
Sample
A549
HEPG2
NMR spectra were recorded in DMSO–d as solvent on
6
6
6
6
6
6
6
6
6
6
a
b
c
d
e
f
g
h
i
0.52
4.9
0.77
0.39
0.31
0.77
0.06
0.04
0.03
0.52
3.72
0.68
9.99
0.73
0.78
0.56
8.9
0.08
0.57
0.16
Varian Gemini NMR spectrometer at 300 MHz using
TMS as internal standard. Chemical shifts are reported as
δ values in ppm. Mass spectra were recorded with a
Shimadzu GCMS–QP–1000 EX mass spectrometer in EI
1.07
2.38
4.03
0.65
0.22
1.19
0.13
(70 eV) model. The elemental analyses were performed
at the Micro analytical center, Cairo University.
Synthesis of compounds 4a and 8. To a mixture of the
appropriate aryl aldehydes 1a or 1b (0.003 mol),
dimedone
3
(0.003 mol) and malononitrile
2
atoms of the carbonyl groups in the two pyran rings with
bond distances 2.5A° and 1.97A°. The fifths binding was
arene–cation interaction between Arg 28 and benzene
ring. The last one is arene–arene interaction between the
amino acid Ph 31 and benzene ring. All these
interactions enhanced the binding activity and fitting of
compound 6g into the active site of the target protein
and hence cancer proliferation inhibition. The data
obtained by the docking program MOE were saved as
PDP file, where the 3D structure of 6g and the 3D
structure of our domains were entered into SCIGRESS
version 3.0 and visualized through BIOVIA Discovery
Studio V6.1.0.15350 (Fig. 3).
(0.0033 mol) in ethanol was added (0.2 mL) as a catalyst,
and the mixture was set at reflux for 3 h. The reaction
mixture was cooled, and the resulting solid was filtered to
afford the crude product, which then recrystallized from
ethanol to give pure 4a or 8.
2-Amino-4,5-dihydro-4-(4-hydroxyphenyl)-5-oxopyrano[3,2-
c]chromene-3-carbonitrile (4a).
Pet ether); Mp = 258–260°C; IR (KBr): ν = 3504, 3407
Off-white solid (EtOAc/
ꢀ
1 1
(br)(NH ), 2197 (s) (CN), 1690 (s) (CO) cm ; H NMR
2
(300 MHz, DMSO-d ): δ 4.29 (s, 1H, CH), 6.65 (d, 2H,
J = 8.1 Hz, Ar-H), 7.01 (d, 2H, J = 8.1 Hz, Ar-H), 7.29
(s, 2H, NH ), 7.38–7.46 (m, 2H, Ar-H.), 7.65 (t, 1H,
J = 7.8 Hz, Ar-H), 7.84 (d, 1H, J = 7.8 Hz, Ar-H), 9.31
6
2
Figure 2. Two-dimensional representation of compound 6g into the active site of human dihydrofolate reductase enzyme, respectively. [Color figure can
be viewed at wileyonlinelibrary.com]
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

A. M. Abdella, M. F. Mohamed, A. F. Mohamed, A. H. M. Elwahy, and I. A. Abdelhamid
Vol 000
Figure 3. Three-dimensional representation of compound 6g into the active sites of human dihydrofolate reductase enzyme, respectively. [Color figure
can be viewed at wileyonlinelibrary.com]
13
(
s, 1H, OH); C NMR (75 MHz, DMSO-d ): δ 36.2, 58.5,
The crude solid was isolated and recrystallized from the
6
1
1
04.6, 113.1, 115.3, 116.6, 119.4, 122.5, 124.7, 128.7,
proper solvent.
0
4
,4 -((Ethane-1,2-diylbis(oxy))bis(2,1-phenylene))bis(2-
32.8, 133.7, 152.1, 153.0, 156.5, 158.0, 159.6, 161.9.
+
amino-5-oxo-4H,5H-pyrano[3,2-c]chromene-3-carbonitrile)
6a). Pale yellow crystals (DMF); Mp = 288–292°C; IR
KBr): ν = 3402 (br)(NH ), 2202 (s) (CN), 1676 (s) (CO)
MS (EI, 70 eV): m/z = 332 [M ], Anal. Calcd for
C H N O : C, 68.67; H, 3.64; N, 8.43%. Found: C,
6
(
19 12 2 4
(
2
8.71; H, 3.66; N, 8.46.
ꢀ1
1
cm ; H NMR (300 MHz, DMSO-d ): δ 3.96–4.14 (m,
6
2
-Oxo-2H-chromene-3-carbonitrile (8).
Off-white solid
4
H, OCH ), 4.627 (s, 2H, pyran H-4), 6.86–7.98 (m,
2
(EtOH-dioxane); Mp = 183–185°C (lit. [19] 184–185°C);
ꢀ
1
1
20H, Ar-H + 2NH
2
). MS (EI, 70 eV): m/z = 690.18
IR (KBr): ν = 2220 (s) (CN), 1712 (s) (CO) cm ; H
NMR (300 MHz, DMSO-d ): δ 7.44–7.75 (m, 4H, Ar-H),
+
[
M ], Anal. Calcd for C H N O : C, 69.56; H, 3.79;
4
0 26 4 8
6
1
3
N, 8.11. Found: C, 69.77; H, 3.56; N, 8.41.
8
1
1
.30 (s, 1H, H4); C NMR (75 MHz, DMSO-d ): δ
03.61, 112.00, 117.56, 117.22, 128.15, 128.21, 136.50,
52.11, 155.20, 156.88. MS (EI, 70 eV): m/z = 171
6
0
4
,4 -((Propane-1,3-diylbis(oxy))bis(2,1-phenylene))bis(2-
amino-5-oxo-4H,5H-pyrano[3,2-c]chromene-3-carbonitrile)
6b). Pale yellow crystals (DMF); Mp = 290–295°C; IR
(KBr): ν = 3430 (br)(NH ), 2195 (s) (CN), 1675 (s) (CO)
(
+
[M ], C H NO : C, 70.18; H, 2.94; N, 8.18. Found (%):
10
5
2
2
ꢀ
1 1
C, 70.23; H, 2.95; N, 8.21.
cm ; H NMR (300 MHz, DMSO-d ): δ 1.94 (br, 2H,
6
General procedure of synthesis of compounds 9a–f [45].
CH ), 3.50–3.86 (m, 4H, OCH ), 4.536 (s, 2H, pyran H-
2
2
To a mixture of bis-aldehydes 7a–f (1 mmol) and
malononitrile 2 (2.2 mmol) in ethanol (20 mL) was added
piperidine (0.2 mL), and the mixture was set at reflux for
4
), 5. 6.17–7.93 (m, 20H, Ar-H + 2NH ). MS (EI,
2
+
7
0
eV): m/z
=
704.19 [M ], Anal. Calcd for
C H N O : C, 69.88; H, 4.01; N, 7.95. Found: C,
41 28 4 8
3
h. The crude solid was isolated and recrystallized from
7
0.05; H, 4.17; N, 8.11.
4,4 -((Butane-1,4-diylbis(oxy))bis(2,1-phenylene))bis(2-
0
the proper solvent.
amino-5-oxo-4H,5H-pyrano[3,2-c]chromene-3-carbonitrile)
6c). Pale yellow crystals (DMF); Mp = 290–293°C; IR
General methods for synthesis of compounds
a–i. Method A. A mixture of bisaldehyde 7 (1 mmol),
malononitrile 2 (2 mmol), and 4-hydroxy-2H-chromen-2-
one 3 (2 mmol) in pyridine (10 mL) was heated at reflux for
5 min. The crude solid was isolated and recrystallized
(
6
(
(
KBr): ν = 3406 (s), 3330 (s) (NH ), 2193 (s) (CN), 1674
2
ꢀ
1 1
s) (CO) cm ; H NMR (300 MHz, DMSO-d ): δ 1.51–
6
1
2
.57 (m, 4H, 2CH ), 3.64–3.85 (m, 4H, 2-OCH ), 4.6 (s,
2
2
1
13
H, pyran H-4), 6.85–7.95 (m, 20H, Ar-H + 2NH2);
): δ 25.5 (CH ), 35.7 (C-4),
-O), 103.2 (C-4a), 112.2 (C6-
C
from the proper solvent.
Method B.
A mixture of bisaldehydes 7 (1 mmol),
NMR (75 MHz, DMSO-d
67.3 (C-3), 90.9 (CH
6
2
malononitrile 2 (2 mmol), and 4-hydroxy-2H-chromen-2-
one 3 (2 mmol) in glacial acetic acid (10 mL) was heated
at reflux in the presence of sodium acetate (3 mmol) for
2
phenylene), 113 (C-10a), 116.4 (C-7), 119.3 (CN), 120.1
(C4-phenylene), 120.9 (C2-phenylene), 123.8 (C-10),
124.5 (C-9), 128.5 (C5-phenylene), 130.2 (C-8), 132.6
(C3-phenylene), 151.9 (C-6a), 153.6 (C1-phenylene),
156.8 (C-2), 158.4 (C-10b), 162.3 (CO). MS (EI, 70 eV):
1
h. The crude solid was isolated and recrystallized from
the proper solvent.
Method C.
A mixture of bis-arylidenemalononitrile
+
derivatives 9a–f (1 mmol) and 4-hydroxy-2H-chromen-2-
one 3 (2.2 mmol) in pyridine was heated at reflux for 3 h.
m/z = 718.21[M ], Anal. Calcd for C H N O : C,
4
2 30 4 8
70.19; H, 4.21; N, 7.80. Found: C, 70.32; H, 4.44; N, 7.58.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2017
Synthesis of Novel bis(dihydropyrano[3,2-c]chromenes)
0
4
,4 -((But-2-ene-1,4-diylbis(oxy))bis(2,1-phenylene))bis(2-
4.76 (s, 2H, pyran H-4), 5.15–5.27 (m, 4H, 2-OCH2),
.87–7.75 (m, 26H, Ar-H + 2NH ). MS (EI, 70 eV):
amino-5-oxo-4H,5H-pyrano[3,2-c]chromene-3-carbonitrile)
6d). Pale yellow crystals (DMF); Mp = 296–300°C; IR
KBr): ν = 3408 (s), 3327 (s) (NH ), 2192 (s) (CN), 1674
s) (CO) cm ; H NMR (300 MHz, DMSO-d ): δ 4.33 (s,
H, 2-OCH ), 4.62 (s, 2H, pyran H-4), 5.72 (s, 2H, vinyl-
6
2
(
(
(
+
m/z = 816.22 [M ], Anal. Calcd for C H N O : C,
50 32 4 8
2
73.52; H, 3.95; N, 6.86. Found: C, 73.32.; H, 3.68; N, 6.97.
ꢀ
1 1
6
Biology. MTT assay.
The MTT assay is performed
4
2
according to Mosmann [46]. The assay was modified for
the cell lines used (A549, MCF7, and HEPG2). Briefly,
these cell lines were exposed to different concentrations of
the tested compounds (10, 5, 2.5, 1.25, 0.62, 0.31, 0.15,
and 0.07 mM), and for the purpose of the experiments at
the end of the incubation time, cells were incubated for 4 h
with 0.8 mg/mL of MTT, dissolved in serum free
mediums. Washing with phosphate-buffered saline (1 mL)
was performed; followed by the addition of DMSO
H), 6.9–7.89 (m, 20H, Ar-H + 2NH ). MS (EI, 70 eV):
2
+
m/z = 716.19 [M ], Anal. Calcd for C H N O : C,
4
2 28 4 8
7
0.39; H, 3.94; N, 7.82. Found: C, 70.52; H, 3.71; N, 7.97.
0
4
,4 -(((1,2-Phenylenebis(methylene))bis(oxy))bis(2,1-
phenylene))bis(2-amino-5-oxo-4H,5H-pyrano[3,2-c]chromene-
-carbonitrile) (6e).
3
Pale yellow crystals (DMF);
Mp = 290–294°C; IR (KBr): ν = 3433 (br) (NH ), 2194
2
ꢀ
1
13
(s) (CN), 1671 (s) (CO) cm
;
C NMR (75 MHz,
DMSO-d ): δ 33.3, 56.8, 67.1, 101.7, 112.1, 112.7,
6
(1 mL), gentle shaking for 10 min so that complete
116.1, 119.4, 120.5, 122.5, 124.3, 128.1, 128.5, 129.1,
dissolution was achieved. Aliquots (200 μl) of the
resulting solutions were transferred in 96-well plates, and
absorbance was recorded at 560 nm using the microplates
spectrophotometer system. Finally, results of cell viability
analysis were analyzed using Prism Software program
1
1
30, 130.3, 132.4, 134.6, 151.8, 153.6, 156.3, 158.1,
1
59.5; H NMR (300 MHz, DMSO-d ): δ 4.62 (s, 2H,
6
pyran H-4), 4.82–4.92 (m, 4H, 2-OCH ), 6.87–7.95 (m,
2
2
4H, Ar-H + 2NH ). MS (EI, 70 eV): m/z = 766.21
2
+
[M ], Anal. Calcd for C H N O : C, 72.06; H, 3.94; N,
46 30 4 8
(
Graphpad Software incorporated, version 3).
Modeling simulation. Docking study for the compound
7
.31. Found: C, 72.26; H, 4.13; N, 7.51.
0
4
,4 -(((1,4-Phenylenebis(methylene))bis(oxy))bis(2,1-
6
g was performed using Molecular Operating Environment
phenylene))bis(2-amino-5-oxo-4H,5H-pyrano[3,2-c]chromene-
-carbonitrile) (6f).
(MOE) version 2009.10 (Chemical Computing Group Inc.,
3
Pale yellow crystals (DMF);
Montreal, QC, Canada) and Autodock4 program. Regular-
ization and optimization for protein and ligand were per-
formed. Determination of the essential amino acids in
binding site was carried out and compared with that present
in literature. The performance of the docking method was
evaluated by redocking crystal ligand into the assigned ac-
tive binding site to determine RMSD value. Docked com-
pound was assigned a score according to its fit in the
ligand binding pocket and its binding mode.
Mp = 298–300°C; IR (KBr): ν = 3419 (br) (NH ), 2195 (s)
2
ꢀ
1
1
(CN), 1674 (s) (CO) cm
;
H NMR (300 MHz,
DMSO-d ): δ 4.75 (s, 2H, pyran H-4), 4.94–5.06 (m, 4H,
6
2
7
-OCH ), 6.88–7.68 (m, 24H, Ar-H + 2NH ). MS (EI,
2 2
+
0 eV): m/z = 766.21 [M ], Anal. Calcd for C H N O :
4
6 30 4 8
C, 72.06; H, 3.94; N, 7.31. Found: C, 72.22; H, 3.73; N, 7.53.
0
4
,4 -(((1,4-Phenylenebis(methylene))bis(oxy))bis(5-bromo-
2,1-phenylene))bis(2-amino-5-oxo-4H,5H-pyrano[3,2-c]
chromene-3-carbonitrile) (6g). Pale yellow crystals (DMF);
Mp >300°C; IR (KBr): ν = 3411 (br) (NH ), 2193 (s) (CN),
2
ꢀ
1 1
1
4
6
672 (s) (CO) cm ; H NMR (300 MHz, DMSO-d ): δ
6
CONCLUSION
.72 (s, 2H, pyran H-4), 4.91–5.05 (m, 4H, 2-OCH2),
.94–7.66 (m, 22H, Ar-H + 2NH ). MS (EI, 70 eV):
2
+
We developed a straightforward methodology for the
preparation of novel bis dihydropyrano[3,2-c]chromenes.
Full characterization of these compounds is reported. The
new synthesized compounds are interesting both in their
own right as unusual molecules and for their promising
pharmacological and biological activities. They offer an
advantage of their simple synthesis in a straightforward
one-stage or two-stage technique from inexpensive
starting materials. Due to the mild reaction conditions,
good yields and selectivity, easily accessible starting
material, and straightforward product isolation, we think
that the new mentioned synthetic approach might offer
new viable techniques for novel bis(functionalized)
heterocycles of expected biological and pharmaceutical
activities. The breast cancer line MCF7 was found to be
the most sensitive one toward most of our derivatives.
Compound 6g may have significant and promising
m/z = 924.03 [M ], Anal. Calcd for C H Br N O : C,
4
6
28
2 4 8
5
9.76; H, 3.05; N, 6.06. Found: C, 59.61; H, 2.88; N, 6.18.
0
4
,4 -(((1,3-Phenylenebis(methylene))bis(oxy))bis(5-bromo-
2,1-phenylene))bis(2-amino-5-oxo-4H,5H-pyrano[3,2-c]
chromene-3-carbonitrile) (6h).
Pale yellow crystals
(
(
(
DMF); Mp = 280–285°C; IR (KBr): ν = 3433 (br)
NH ), 2193 (s) (CN), 1671 (s) (CO) cm ; H NMR
300 MHz, DMSO-d ): δ 4.70 (s, 2H, pyran H-4), 4.80–
ꢀ
1
1
2
6
4
.94 (m, 4H, 2-OCH ), 6.95–7.65 (m, 22H, Ar-
2
+
H + 2NH ). MS (EI, 70 eV): m/z = 924.03 [M ], Anal.
Calcd for C H Br N O : C, 59.76; H, 3.05; N, 6.06.
Found: C, 59.91; H, 2.84; N, 6.27.
2
46
28
2 4 8
0
4
,4 -(((Naphthalene-2,6-diylbis(methylene))bis(oxy))bis(2,1-
phenylene))bis(2-amino-5-oxo-4H,5H-pyrano[3,2-c]chromene-
-carbonitrile) (6i).
3
>
Pale yellow crystals (DMF); Mp
300°C; IR (KBr): ν = 3403 (br) (NH ), 2195 (s) (CN),
2
ꢀ
1 1
1
672 (s) (CO) cm ; H NMR (300 MHz, DMSO-d ): δ
6
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

A. M. Abdella, M. F. Mohamed, A. F. Mohamed, A. H. M. Elwahy, and I. A. Abdelhamid
Vol 000
growth inhibitory efficiency against the three cell lines
especially MCF7. Docking studies of compounds 6g
showed the best binding mode with the target protein.
[22] Dolle, R. E. J Comb Chem 2004, 6, 623.
23] Helal, C. J.; Sanner, M. A.; Cooper, C. B.; Gant, T.; Adam, M.;
Lucas, J. C.; Kang, Z.; Kupchinsky, S.; Ahlijanian, M. K.; Tate, B.;
Menniti, F. S.; Kelly, K.; Peterson, M. Bioorg Med Chem Lett 2004, 14,
[
5521.
[24] Soural, M.; Bouillon, I.; Krchňák, V. J Comb Chem 2008, 10,
923.
REFERENCES AND NOTES
[
[
25] Shaker, R. M. Pharmazie 1996, 51, 148.
26] Abdolmohammadi, S.; Balalaie, S. Tetrahedron Lett 2007, 48,
[
1] Tanabe, A.; Nakashima, H.; Yoshida, O.; Yamamoto, N.;
Tenmyo, O.; Oki, T. J Antibiot 1988, 41, 1708.
2] Sangani, C. B.; Mungra, D. C.; Patel, M. P.; Patel, R. G. Cent
Eur J Chem 2011, 9, 635.
3] El-Saghier, A. M. M.; Naili, M. B.; Rammash, B. K.; Saleh, N.
A.; Kreddan, K. M. ARKIVOC 2007, xvi, 83.
3
299.
[
[
[
[
27] Kidwai, M.; Saxena, S. Synth Commun 2006, 36, 2737.
28] Seifi, M.; Sheibani, H. Catal Lett 2008, 126, 275.
[
29] Khurana, J. M.; Kumar, S. Tetrahedron Lett 2009, 50, 4125.
30] Heravi, M. M.; Jani, B. A.; Derikvand, F.; Bamoharram, F. F.;
[
Oskooie, H. A. Cat Com 2008, 10, 272.
31] Wang, H.-J.; Lu, J.; Zhang, Z.-H. Monatshefte Für Chemie -
Chem Mon 2010, 141, 1107.
32] Shaabani, A.; Samadi, S.; Badri, Z.; Rahmati, A. Catal Lett
005, 104, 39.
33] Ghorbani-Vaghei, R.; Toghraei-Semiromi, Z.; Karimi-Nami,
R. J Braz Chem Soc 2011, 22, 905.
[
[
4] Ganem, B. Acc Chem Res 2009, 42, 463.
5] Dömling, A.; Wang, W.; Wang, K. Chem Rev 2012, 112,
[
3
083.
[
[
6] Jieping Zhu, H. B. Multicomponent Reactions; John Wiley &
2
Sons: China, 2006.
[
[
7] Ruijter, E.; Scheffelaar, R.; Orru, R. V. A. Angew Chem Int Ed
Engl 2011, 50, 6234.
[
[
[
34] Shaterian, H. R.; Oveisi, A. R. J Iran Chem Soc 2012, 8, 545.
35] Shaterian, H. R.; Arman, M.; Rigi, F. J Mol Liq 2011, 158, 145.
36] Dekamin, M. G.; Eslami, M.; Maleki, A. Tetrahedron 2013,
[
[
8] Gore, R. P.; Rajput, A. P. Drug Invent Today 2013, 5, 148.
9] Dommaraju, Y.; Bora, S.; Prajapati, D. Org Biomol Chem
2
015, 13, 9181.
10] Ramazani, A.; Khoobi, M.; Torkaman, A.; Zeinali Nasrabadi,
F.; Forootanfar, H.; Shakibaie, M.; Jafari, M.; Ameri, A.; Emami, S.;
Faramarzi, M. A.; Foroumadi, A.; Shafiee, A. Eur J Med Chem 2014,
8, 151.
6
1
3
9, 1074.
[
[37] Mehrabi, H.; Kazemi-Mireki, M. Chin Chem Lett 2011, 22,
419.
[38] Shaterian, H. R.; Honarmand, M. Synth Commun 2011, 41,
7
4
8
4
4
573.
[
[
[
[
[
11] Elwahy, A. H. M.; Shaaban, M. R. Curr Org Synth 2010, 7,
[39] Zhang, M.; Zhang, A. Synth Commun 2004, 34, 4531.
33.
35.
25.
71.
[40] Valizadeh, H.; Gholipur, H.; Shockravi, A. J Heterocyclic
12] Elwahy, A. H. M.; Shaaban, M. R. Curr Org Synth 2015, 11,
13] Elwahy, A. H. M.; Shaaban, M. R. Curr Org Synth 2015, 10,
14] Shaaban, M. R.; Elwahy, A. H. M. Curr Org Synth 2015, 11,
15] Wang, C.; Jung, G.-Y.; Hua, Y.; Pearson, C.; Bryce, M. R.;
Chem 2007, 44, 867.
41] Sajadikhah, S. S.; Maghsoodlou, M. T.; Hazeri, N.; Norouzi,
M.; Moein, M. Res Chem Intermed 2015, 41, 8665.
42] Elwahy, A. H. M. J Chem Res 1999, (S) 602.
[
[
[43] Muathen, H. A.; Aloweiny, N. A. M.; Elwahy, A. H. M. J
Heterocyclic Chem 2009, 46, 656.
[44] Ibrahim, Y. A.; Elwahy, A. H. M.; Elkareis, G. M. M. J Chem
Petty, M. C.; Batsanov, A. S.; Goeta, A. E.; Howard, J. A. K. Chem Mater
001, 13, 1167.
16] Wang, C.; Jung, G.-Y.; Batsanov, A. S.; Bryce, M. R.; Petty,
M. C. J Mater Chem 2002, 12, 173.
17] Thurston, D. E.; Bose, D. S.; Thompson, A. S.; Howard, P. W.;
Res 1994, (S) 414.
2
[
[45] Salama, S. K.; Darweesh, A. F.; Abdelhamid, I. A.; Elwahy, A.
H. M. J Heterocyclic Chem 2017, 54, 305.
[46] Mosmann, T. J Immunol Methods 1983, 65, 55.
[
Leoni, A.; Croker, S. J.; Jenkins, T. C.; Neidle, S.; Hartley, J. A.; Hurley,
L. H. J Org Chem 1996, 61, 8141.
[
18] Yang, G. Y.; Oh, K.-A.; Park, N.-J.; Jung, Y.-S. Bioorg Med
Chem 2007, 15, 7704.
19] Holla, B. S.; Gonsalves, R.; Shenoy, S. Eur J Med Chem 2000,
5, 267.
SUPPORTING INFORMATION
[
3
Additional Supporting Information may be found online
in the supporting information tab for this article.
[
[
20] Murru, S.; Nefzi, A. ACS Comb Sci 2014, 16, 39.
21] Dolle, R. E. J Comb Chem 2005, 7, 739.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet