Synthesis and Anti-influenza Virus Activity of Novel bis(4H-chromene-3-carbonitrile) Derivatives

Article abstract of DOI:10.1002/jhet.2776

An efficient and convenient method for the synthesis of bis(4H-chromene-3-carbonitrile) derivatives by one-pot, multicomponent reaction of bis-aldehydes, malononitrile, and dimedone in the presence of a catalytic amount of piperidine is reported. Bis(2-benzylidene-1H-indene)-1,3-(2H)-dione derivatives were obtained as the main products as a result of reaction of the bis(arylidenemalononitriles) with indandione. The anti-influenza H5N1 virus activities of the newly prepared bis-chromene derivatives are also investigated.

Full text of DOI:10.1002/jhet.2776

Month 2016 Synthesis and Anti-influenza Virus Activity of Novel bis(4H-chromene-3-
carbonitrile) Derivatives
Amna M. Abdella, Yassmin Moatasim, Mohamed A. Ali, Ahmed H. M. Elwahy, * and Ismail A. Abdelhamid *
a
b
b
a
a
a
Chemistry Department, Faculty of Science, Cairo University, Giza, Egypt
b
Center of Scientific Excellence for Influenza Viruses, National Research Centre, Giza, Egypt
*
E-mail: aelwahy@hotmail.com; ismail_shafy@yahoo.com
Received March 12, 2016
DOI 10.1002/jhet.2776
Published online 00 Month 2016 in Wiley Online Library (wileyonlinelibrary.com).
An efficient and convenient method for the synthesis of bis(4H-chromene-3-carbonitrile) derivatives by
one-pot, multicomponent reaction of bis-aldehydes, malononitrile, and dimedone in the presence of a
catalytic amount of piperidine is reported. Bis(2-benzylidene-1H-indene)-1,3-(2H)-dione derivatives were
obtained as the main products as a result of reaction of the bis(arylidenemalononitriles) with indandione.
The anti-influenza H5N1 virus activities of the newly prepared bis-chromene derivatives are also
investigated.
J. Heterocyclic Chem., 00, 00 (2016).
INTRODUCTION
causing global concern as a potential pandemic threat.
Two classes of antiviral agents are recommended
as drugs for treatment of influenza virus infection
in humans: adamantanes (M2 blockers) [30] and
neuraminidase inhibitors [31]. Some evidences of
antiviral drug resistance have been recorded in H5N1
viruses isolated from some human cases [32,33]. So the
screening and developing of new classes of antiviral
drugs is a significant and an urgent task.
The Michael addition reactions of α,β-unsaturated
nitriles with β-diketones is an interesting route for the
synthesis of chromene and fused chromene derivatives
[
1–10]. Chromene derivatives are an important class of
compounds of considerable interests, which have been
the subject of extensive research, especially in the
pharmaceuticals, including anticancer [11–15], antiviral
[
16–18] antimicrobial [19,20], and anti-inflammatory
As a part of an ongoing research program on Michael
addition reactions [34–42] and bis-heterocyclic
activities [21]. Some chromene derivatives were found to
have potential ability in the enhancement of cognitive
functions, and thus they are used in the treatment of
neurodegenerative diseases [15]. 2-Amino-4H-chromene
derivatives bearing nitrile functionality showed potential
application in the treatment of human inflammatory
TNFα-mediated diseases, such as rheumatoid and
psoriatic arthritis [22,23].
compounds with
a
suitable spacer [43–52], and
encouraged by the potency of the clinically useful
chromene derivatives in treatment of cancer and
inflammation and other activities, we report the results of
our investigations concerning the synthesis of novel
bis-4H-chromene-3-carbonitrile derivatives aiming at
exploring of their anti-influenza virus activities.
Chromene derivatives play also an important role in the
production of highly effective fluorescent dyes for daylight
fluorescent pigments and synthetic fibers [24]. In addition,
bis-heterocyclic compounds with a suitable spacer are
reported to exhibit various pharmacological activities
including, antitumor [25] and antimicrobial activities
RESULTS AND DISCUSSION
The synthetic strategy used in the synthesis of the
target bis-4H-chromene-3-carbonitrile molecules is
outlined in scheme 1. Thus, a variety of new bis-
arylidenemalononitrile derivatives 3 were obtained in
80–90% yields via Knoevenagel condensation of one
mole of bis-aldehyde derivatives 1 with two moles
of malononitrile 2 in ethanol and in the presence of
piperidine as a basic catalyst. In the next step, one mole
of the isolated bis-arylidenemalononitrile derivatives 3
was reacted with two moles of dimedone 4, in the
[26]. Moreover, recent reports showed that among
libraries of derivatized heterocycles, the most active
library compounds had a bis-heterocyclic structure [27,28].
Seasonal human influenza is one of the most common
respiratory infection viruses, affecting millions of people
worldwide every year [29]. In recent years, the highly
pathogenic avian influenza H5N1 virus crossed the
species barrier and transmitted to human that has been
©
2016 Wiley Periodicals, Inc.

A. M. Abdella, Y. Moatasim, M. A. Ali, A. H. M. Elwahy, and I. A. Abdelhamid
Vol 000
Scheme 1. Synthesis of bis(4H-chromene-3-carbonitrile) derivatives by a step-wise approach.
presence of piperidine via Michael addition reaction to
yield 5 in good yields (Scheme 1).
The reaction proceeds most likely via initial Michael
addition of dimedone 4 to the C= C bond in 3, followed
by cyclization to give the final isolable products 5
through intermediacy of 6–8 (Scheme 2).
Compounds 5a–h could also be alternatively obtained in
a multicomponent reaction of one mole of bis-aldehydes 1
with two moles of both malononitrile and dimedone in
EtOH/piperidine (Scheme 3).
On the other hand, attempts to synthesize bis(4H-
chromene-3-carbonitrile) derivatives 5 by bis-alkylation of
4H-chromene-3-carbonitrile 9 with the appropriate dihalo
compounds 10 in basic medium were unfortunately
unsuccessful (Scheme 4). The reaction gave instead
a
mixture of products that were not easily
separated and have not been characterized yet. The
monopodal 4H-chromene-3-carbonitrile 9 was synthesized
via multicomponent reaction of the appropriate
hydroxybenzaldehyde, malononitrile, and dimedone in
EtOH in the presence of piperidine as a catalyst [53,54].
The constitutions of compounds 5 were established
spectroscopically based on the elemental analysis and
spectral data. The IR spectra of compound 5a as a
representive example indicated the presence of amino
2
-amino-7,7-dimethyl-5-oxo-4-phenyl-5,6,7,8-tetrahydro-
Scheme 2. Proposed mechanism for the formation of bis(4H-chromene-3-carbonitrile) derivatives.
Journal of Heterocyclic Chemistry
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Anti-influenza Virus Activity of bis(4H-chromene-3-carbonitrile) Derivatives
Scheme 3. Synthesis of bis(4H-chromene-3-carbonitrile) derivatives by multicomponent reactions.
Scheme 4. Attempted synthesis of bis(4H-chromene-3-carbonitrile)
derivatives by a bis-alkylation strategy.
ether linkage OCH as multiplet or two separate doublet
2
signals at 3.98–4.30ppm, although their precursors 3a,c,
e,g,h exhibit singlet or triplet signals for these protons.
This suggests that the generated asymmetric center (in the
dihydropyran rings) is close enough to this CH group.
2
On the other hand, the two methylene ether linkage
OCH resonance appears as a singlet in compounds 5b,d,
2
f, which indicate that the asymmetric center is not close
enough to effect such splitting.
On the other hand, repeated attempts to
prepare bis-dihydroindeno[1,2-b]pyran-3-carbonitrile 13,
by the reaction of one mole of bis-aldehydes 1 with
two moles of both malononitrile and indanedione
(11) instead of dimedone under similar reaction
conditions were unsuccessful. Instead, the reaction
group at 3435 and 3350. In addition, it revealed the cyano
ꢀ
1
group at 2194 cm . The carbonyl group appeared as a
ꢀ
1
1
broad band at 1668cm . The H-NMR spectrum of 5a
indicated the presence of two singlets integrated by 12
protons at δ 0.95 and 1.06 ppm assigned to four CH . In
3
addition, it indicated two characteristic doublets of
doublet at 2.11 and 2.25 ppm with coupling constant
J =16.1 ppm assigned to H8. The singlet signal at
afforded
bis(methanylylidene))bis(1H-indene-1,3(2H)-
dione) derivatives 14a–c as single products in good yield
(Scheme 5). The structure of compounds 14a–c was
confirmed by comparison with their physical data with
authentic samples synthesized from condensation of one
mole of bis-aldehyde derivatives 1 with two moles of
2.50ppm is assigned to H6. The pyran-H4 appeared as a
singlet signal at δ= 4.50ppm. Moreover, compound 5a as
well as compounds 5c,e,g,h also featured the methylene
Scheme 5. Attempted synthesis of bis-dihydroindeno[1,2-b]pyran-3-carbonitriles by multicomponent reactions.
Journal of Heterocyclic Chemistry
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A. M. Abdella, Y. Moatasim, M. A. Ali, A. H. M. Elwahy, and I. A. Abdelhamid
Vol 000
Figure 1. TC50 of synthetic compounds. [Color figure can be viewed at wileyonlinelibrary.com.]
indanedione 11 in EtOH in the presence of piperidine as a
basic catalyst. The formation of 14 is assumed to proceed
via initial formation of the adduct 12, which then
decompose to give 14 via elimination of malononitrile. A
similar sequence has been reported [34,55–60].
showed that most of tested compounds produced
moderate antiviral activity (50% inhibition) against tested
influenza virus as shown in Table 2. It has been noticed
that the best antiviral potency among tested compounds
was obtained by compounds 5b and 5g compounds that
showed 76.1% and 77.2% antiviral inhibition, respectively.
The constitutions of compounds 14 were established
1
spectroscopically. For example, the H-NMR spectrum of
compound 14a revealed singlet signal integrated by 4H at
δ 4.60 ppm assigned the two –OCH groups. The ylidene
2
Table 1
H-atoms appeared as singlet signal at 8.25ppm. In
addition, it revealed the aromatic protons as multiplets in
their expected positions.
TC50 of synthetic compounds
Compound
TC50 Conc. (μM/μl)
Antiviral Evaluation. The in vitro antiviral efficacy of
the newly synthesized bis(4H-chromene-3-carbonitrile)
derivatives against highly pathogenic avian influenza
H5N1 virus was screened in MDCK cells. The
concentrations of the tested compounds, which exhibited
5a
67.7
696.15
410.2
132.1
63.48
638.508
43.5
5
5
5
5
b
c
d
e
5
0% cytotoxicity TC50, were determined (Fig. 1 and
5f
5
5
g
h
Table 1). The obtained results of plaque reduction assay
at two safe concentrations (<TC50) of each compound
26.674
Journal of Heterocyclic Chemistry
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Anti-influenza Virus Activity of bis(4H-chromene-3-carbonitrile) Derivatives
Table 2
Antiviral activity of the tested compounds against H5N1 virus
2,2'-(((Ethane-1,2-diylbis(oxy))bis(2,1-phenylene))bis
(methanylylidene))-dimalononitrile (3a).
Pale yellow
1
crystals (dioxane), Mp = 243–245°C.[45]
H-NMR
Compound
Concentration (μM)
% of virus inhibition
(
(
300 MHz, DMSO-d ): δ= 4.6 (s, 4H, OCH ), 7.16–8.05
m, 8H, Ar-H), 8.43 (s, 2H, vinyl-H).
6
2
5
5
5
5
5
5
5
5
a
b
c
16.9373
43.9
0
76.1
65
8
.46864
174.038
7.0192
205.13
02.565
132.104
6.06
15.8717
.93584
159.627
9.8135
32.625
1.75
13.3373
.66866
2,2'-(((Ethane-1,2-diylbis(oxy))bis(4,1-phenylene))bis
methanylylidene))-dimalononitrile (3b).
(
Pale yellow
8
1
crystals (dioxane), Mp = 162–164°C.[61]
H-NMR
54.5
51.5
62.5
0
52.7
13.8
68.4
47.3
77.27
65.21
45.4
0
1
(300 MHz, DMSO-d6): δ= 4.51 (s, 4H, 2OCH2).
13
d
e
C-NMR (75 MHz, DMSO-d ): δ= 66.7, 77.1, 113.8,
6
6
1
14.7, 124.4, 133.3, 160.3, 163.2.
2
,2'-(((Propane-1,3-diylbis(oxy))bis(2,1-phenylene))bis
7
f
(methanylylidene))-dimalononitrile (3c).
Pale yellow
1
7
crystals (dioxane / ethanol), Mp = 214–216°C.[45] H-
g
h
NMR (300MHz, DMSO-d ): δ= 2.35 (quintet, 2H,
6
2
OCH ), 4.36 (t, 4H, OCH ), 7.13–8.03 (m, 8H, ArH's),
2
2
6
8.52 (s, 2H, vinyl-H)
2
,2'-(((Propane-1,3-diylbis(oxy))bis(4,1-phenylene))bis
(
methanylylidene))-dimalononitrile (3d).
Pale yellow
1
CONCLUSIONS
crystals (dioxane/ethanol), Mp = 170–172°C.[54] H-
NMR (300MHz, DMSO-d ): δ= 2.25 (q, 2H, J = 6 Hz,
6
We have developed an efficient synthesis of novel
bis-4H-chromene-3-carbonitrile derivatives via the
Michael addition reaction of dimedone towards bis-
arylidenemalononitrile derivatives. The new synthesized
compounds offer an advantage of their easy synthesis in
a simple one-pot or stepwise procedure from inexpensive
starting materials. Our current studies are directed to
extend the scope of this method to cover additional
bis-heterocyclic compounds. The anti-influenza activities
of the newly synthesized bis-heterocyclic compounds
were investigated, and the best antiviral potency among the
tested compounds was obtained by compounds 5b and 5g.
OCH CH ), 4.29 (t, 4H, J = 6 Hz, 2OCH ), 7.20 (d, 4H,
2
2
2
J = 9Hz, Ar-H), 7.96 (d, 4H, J =9 Hz, Ar-H), 8.37 (s, 2H,
13
vinyl-H). C-NMR (75 MHz, DMSO-d ): δ= 28.1, 64.9,
6
76.9, 113.8, 114.7, 124.1, 133.3, 160.3, 163.5.
2,2'-(((Butane-1,4-diylbis(oxy))bis(2,1-phenylene))bis
(methanylylidene))-dimalononitrile (3e).
Pale yellow
crystals (dioxane), Mp = 210–212°C, IR (KBr): ν = 2190
ꢀ
1 1
(CN) cm . H-NMR (300MHz, DMSO-d ): δ= 1.90 (s,
6
4H, 2 CH ), 4.22 (s, 4H, 2-OCH ), 7.11–7.99 (m, 8H,
2
2
13
Ar-H), 8.41 (s, 2H, vinyl-H).
C-NMR (75 MHz,
DMSO-d ): δ= 24.9, 67.9, 76.7, 113.9, 114.8, 115.6,
6
120.5, 124.0, 133.3, 160.4, 163.7. MS (EI, 70eV): m/z
+
(
%) = 394 [M ], Anal. Calcd for C H N O : C, 73.08;
2
4 18 4 2
H, 4.60; N, 14.20. Found: C, 73.19; H, 4.67; N, 14.31.
EXPERIMENTAL
2,2'-(((Butane-1,4-diylbis(oxy))bis(4,1-phenylene))bis
(
methanylylidene))-dimalononitrile (3f).
Pale yellow
General. Melting points were measured with a Stuart
melting point apparatus and are uncorrected. The IR
spectra were recorded using a FTIR Bruker–vector 22
crystals (82%), (dioxane/ethanol), Mp = 205–207°C, IR
ꢀ
1
1
(KBr): ν = 2191 (CN) cm
.
H-NMR (300MHz,
DMSO-d ): δ= 1.91 (br, 4H, 2 CH ), 4.19 (br, 4H, 2-
6
2
1
spectrophotometer as KBr pellets. The H-NMR spectra
OCH ), 7.19 (d, 4H, Ar-H, J= 9 Hz), 7.97 (d, 4H, Ar-H,
2
13
were recorded in DMSO–d as solvent on Varian Gemini
J = 9Hz), 8.37 (s, 2H, vinyl-H). C-NMR (75 MHz,
DMSO-d6): δ =24.9, 66.8, 76.7, 113.8, 114.7, 124.0,
6
NMR spectrometer at 300MHz using TMS as internal
standard. Chemical shifts are reported as δ values in
ppm. Mass spectra were recorded with a GCMS–QP–
+
133.3, 160.4, 163.7. MS (EI, 70eV): m/z (%) = 394 [M ],
Anal. Calcd for C H N O : C, 73.08; H, 4.60; N,
24 18 4 2
1
000 EX mass spectrometer (Shimadzu, Japan) in EI
14.20. Found: C, 73.29; H, 4.72; N, 14.23.
(70 eV) model. The elemental analyses were performed at
2,2'-(((Ethane-1,2-diylbis(oxy))bis(5-bromo-2,1-phenylene))
the Microanalytical Center, Cairo University.
bis-(methanylylidene))dimalononitrile (3g).
Pale yellow
General Procedure for the Synthesis of Compounds 3a–h.
To a mixture of bis-aldehydes 1a–h (1mmol) and
malononitrile 2 (2.2mmol) in ethanol (20mL) piperidine
crystals (87%), (dioxane/ethanol), mp 258–260°C, IR
ꢀ
1
1
(KBr): ν= 2195 (CN) cm
,
H-NMR (300 MHz,
DMSO-d ): δ= 4.56 (s, 4H, 2OCH ), 7.31 (d, 2H, Ar-H,
6
2
(0.2mL) was added. The mixture was heated under reflux
J = 8.7 Hz), 7.82 (d, 2H, Ar-H, J = 8.7 Hz), 8.09 (s, 2H,
Ar-H), 8.42 (s, 2H, vinyl-H). MS (EI, 70 eV): m/z (%)
for 3h. The crude solid was isolated and recrystallized
from the proper solvent.
+
= 524 [M ], Anal. Calcd for C H Br N O C, 50.41; H,
22
12
2 4 2
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A. M. Abdella, Y. Moatasim, M. A. Ali, A. H. M. Elwahy, and I. A. Abdelhamid
Vol 000
2
3
.31; Br, 30.49; N, 10.69. Found: C, 50.59; H, 2.43; Br,
0.39; N, 10.54.
4,4'-((Propane-1,3-diylbis(oxy))bis(2,1-phenylene))bis(2-
amino-7,7 dimethyl-5-oxo-5,6,7,8-tetrahydro-4H-chromene-3-
carbonitrile) (5c). Pale yellow crystals (Method A, 80%,
Method B, 84%), (dioxane / ethanol), Mp=240–242°C,
2
,2'-(((Butane-1,4-diylbis(oxy))bis(5-bromo-2,1-phenylene))
bis-(methanylylidene))dimalononitrile (3h). Pale yellow
crystals (83%), (dioxane/ethanol), mp 268–270°C, IR
IR (KBr): ν = 3396, 3305 (NH ), 2186 (CN), 1654 (CO)
2
ꢀ
1
1
ꢀ1
1
(
KBr): ν = 2188 (CN) cm
,
H-NMR (300 MHz,
cm . H-NMR (300MHz, DMSO-d ): δ =0.93 (s, 6H,
6
DMSO-d ): δ= 1.96 (s, 4H, 2CH ), 4.21 (s, 4H, 2OCH ),
2CH ), 1.04 (s, 6H, 2CH ), 2.0–2.25 (m, 6H, H8+
6
2
2
3
3
7
7
.20–8.07 (m, 6H, Ar-H), 8.35 (s, 2H, vinyl-H). MS (EI,
0eV): m/z (%) = 549 [M ], Anal. Calcd for
OCH CH CH O), 2.50 (s, 4H, H6), 4.03–4.21 (m, 4H,
2 2 2
+
2OCH ), 4.46 (s, 2H, pyran H-4) 6.81–7.13 (m, 12H,
2
13
C H Br N O C, 52.20; H, 2.92; Br, 28.94; N, 10.15.
ArH+2NH2). C-NMR (75MHz, DMSO-d ): δ=26.4,
24
16
2
4
2
6
Found: C, 52.16; H, 2.81; Br, 28.83; N, 10.28.
28.5, 28.9, 31.4, 31.7, 50.1, 57.1, 64.8, 111.9, 119.9,
General Procedure for the Synthesis of Compounds 5a–h.
Method A: A mixture of bis-arylidenemalononitrile
derivatives 3a–h (1mmol) and dimedone 4 (2.2mmol) in
absolute ethanol (15mL) was heated at reflux in the
presence of piperidine (0.2mL) for 3h. The crude solid
was isolated and recrystallized from the proper solvent.
Method B: To a mixture of bisaldehydes 1a–i (1mmol),
malononitrile 2 (2.2 mmol), and dimedone 4 (2.2 mmol)
in absolute ethanol (15 mL), piperidine (0.2 mL) was
added, and the reaction mixture was heated at reflux for
120.0, 120.6, 127.9, 129.3, 131.5, 156.3, 158.9, 162.7,
195.5. MS (EI, 70eV): m/z (%)=660 [M ]. Anal. Calcd
+
for C H N O : C, 70.89; H, 6.10; N, 8.48. Found: C,
39 40 4 6
70.93; H, 6.02; N, 8.51.
4,4'-((Propane-1,3-diylbis(oxy))bis(4,1-phenylene))bis(2-
amino-7,7-dimethyl-5-oxo-5,6,7,8-tetrahydro-4H-chromene-3-
carbonitrile) (5d). Pale yellow crystals (Method A, 81%,
Method B, 79%), (dioxane/ethanol), Mp=140–142°C, IR
ꢀ
1 1
(KBr): ν =3331 (NH ), 2191 (CN), 1679 (CO) cm . H-
2
NMR (300 MHz, DMSO-d ): δ =0.94 (s, 6H, 2CH ), 1.03
6
3
3
h. The crude solid was isolated and recrystallized from
(s, 6H, 2CH ), 1.98–2.26 (m, 6H, H8+ OCH CH CH O),
3 2 2 2
the proper solvent.
2.50 (s, 4H, H6), 4.02–4.11 (m, 6H, 2OCH + pyran H-4)
6.84–7.05 (m, 12H, ArH + 2NH2). C-NMR (75MHz,
2
13
4
,4'-((Ethane-1,2-diylbis(oxy))bis(2,1-phenylene))bis(2-amino-
,7-dimethyl-5-oxo-5,6,7,8-tetrahydro-4H-chromene-3-carbonitrile)
5a). Pale yellow crystals (Method A, 84%; Method B,
7
(
DMSO-d ): δ=26.7, 28.6, 31.7, 34.7, 50.0, 58.6, 64.2,
6
112.9, 114.2, 119.7, 128.2, 136.8, 157.2, 158.4, 162.1,
+
8
7%), (dioxane / ethanol), Mp = 277–279°C, IR (KBr):
195.6. MS (EI, 70eV): m/z (%)=660 [M ]. Anal. Calcd
ꢀ
1
ν = 3435, 3350 (NH ), 2194 (CN), 1668 (CO) cm
.
for C H N O : C, 70.89; H, 6.10; N, 8.48. Found: C,
2
39 40 4 6
1
H-NMR (300 MHz, DMSO-d ): δ= 0.95 (s, 6H,
70.95; H, 6.13; N, 8.58.
6
2
CH ), 1.06 (s, 6H, 2CH ), 2.11 (d, 2H, H8,
4,4'-((Butane-1,4-diylbis(oxy))bis(2,1-phenylene))bis(2-amino-
7,7-dimethyl-5-oxo-5,6,7,8-tetrahydro-4H-chromene-3-carbonitrile)
(5e). Pale yellow crystals (Method A, 80%, Method B,
83%), (dioxane/ethanol), Mp = 215–217°C. IR (KBr):
3
3
J = 16.2 Hz), 2.25 (d, 2H, H8, J = 16.2 Hz), 2.50 (s, 4H,
H6), 4.27–4.30 (m, 4H, 2OCH ), 4.50 (s, 2H, pyran
H-4) 6.86–7.22 (m, 12H, ArH + 2NH2).
2
1
3
C-NMR
ꢀ
1 1
(
75 MHz, DMSO-d6): δ= 26.6, 28.4, 31.8, 38.9, 50.1,
ν = 3326 (NH ), 2191 (CN), 1660 (CO) cm . H-NMR
2
5
1
7.1, 66.3, 111.7, 112.7, 119.9, 120.7, 127.9, 129.2,
(300 MHz, DMSO-d ): δ = 0.96 (s, 6H, 2CH ), 1.06
6
3
32.1, 156.1, 159.0, 162.8, 195.6. MS (EI, 70 eV): m/z
(s,
6H,
2CH3),
1.94–2.45
(m,
8H,
H8
+
(
%) = 646 [M ], Anal. Calcd for C H N O :
+ OCH CH CH CH O), 2.50 (s, 4H, H6), 3.98–4.05
3
8
38
4
6
2 2 2 2
C, 70.57; H, 5.92; N, 8.66. Found: C, 70.63; H, 5.87;
N, 8.83.
(m, 4H, 2OCH ) 4.49 (s, 2H, pyran H-4), 6.77–7.17
2
(m, 12H, ArH + 2NH ). MS (EI, 70 eV): m/z (%) = 674
2
+
4
,4'-((Ethane-1,2-diylbis(oxy))bis(4,1-phenylene))bis(2-amino-
,7-dimethyl-5-oxo-5,6,7,8-tetrahydro-4H-chromene-3-carbonitrile)
5b). Pale yellow crystals (Method A, 81%, Method B,
[M ]. Anal. Calcd for C H N O : C, 71.20; H, 6.27;
4
0
42
4
6
7
(
N, 8.30. Found: C, 71.31; H, 6.36; N, 8.27.
4,4'-((Butane-1,4-diylbis(oxy))bis(4,1-phenylene))bis(2-amino-
7,7-dimethyl-5-oxo-5,6,7,8-tetrahydro-4H-chromene-3-carbonitrile)
(5f). Pale yellow crystals (Method A, 74%, Method B,
77%), (dioxane/ethanol), Mp = 187–190°C, IR (KBr):
8
3%), (dioxane/ethanol), Mp = 163-165°C, IR (KBr):
ꢀ
1
ν =3386, 3327 (NH ), 2191 (CN), 1679 (CO) cm
.
2
1
H-NMR (300 MHz, DMSO-d ): δ = 0.94 (s, 6H, 2CH ),
6
3
ꢀ
1
1
.03 (s, 6H, 2CH ), 2.06 (d, 2H, H8, J = 15.9 Hz), 2.26
ν = 3400 3327 (NH ), 2193 (CN), 1662 (CO) cm .
3
2
1
(d, 2H, H8, J = 15.9 Hz), 2.50 (s, 4H, H6), 4.12 (s, 2H,
H-NMR (300 MHz, DMSO-d ): δ = 0.95 (s, 6H, 2CH ),
6 3
pyran H-4), 4.24 (s, 4H, 2OCH ), 6.86 (s, 4H, 2 NH ),
1.03 (s, 6H, 2CH ), 1.83 (s, 4H, OCH CH CH CH O),
2
2
3
2
2
2
2
6
.90 (d, 4H, Ar-H, J =8.7 Hz), 7.07 (d, 4H, Ar-H,
2.06 (d, 2H, H8, J = 15.9 Hz), 2.27 (d, 2H, H8,
13
J = 8.7 Hz);
C-NMR (75 MHz, DMSO-d ): δ = 26.8,
J = 15.9 Hz), 2.50 (s, 4H, H6), 3.98 (s, 4H, 2OCH ) 4.11
6
2
2
1
8.4, 31.8, 34.8, 50.0, 58.5, 66.3, 112.9, 114.2, 119.8,
(s, 2H, pyran H-4), 6.82–7.05 (m, 12H, ArH + 2NH ).
2
1
3
28.3, 137.1, 157.0, 158.4, 162.1, 195.6. MS (EI, 70eV):
C-NMR (75 MHz, DMSO-d ): δ= 25.4, 26.7, 28.3,
6
+
m/z (%) = 646 [M ], Anal. Calcd for C H N O : C,
31.7, 34.7, 49.9, 58.6, 67.0, 112.9, 114.2, 119.7, 128.1,
3
8 38 4 6
7
0.57; H, 5.92; N, 8.66. Found: C, 70.62; H, 5.89; N, 8.80.
136.7, 157.3, 158.4, 162.0, 195.5. MS (EI, 70 eV): m/z
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2016
Anti-influenza Virus Activity of bis(4H-chromene-3-carbonitrile) Derivatives
+
ꢀ1
(
6
%) = 674 [M ]. Anal. Calcd for C H N O : C, 71.20; H, 90%), Mp 248–250°C. IR (KBr): ν = 1667 (CO) cm
.
4
0 42 4 6
1
.27; N, 8.30. Found: C, 71.29; H, 6.38; N, 8.29.
H-NMR (300MHz, DMSO-d ): δ= 4.60 (s, 4H, 2OCH )
6 2
4
,4'-((Ethane-1,2-diylbis(oxy))bis(5-bromo-2,1-phenylene))bis
7.1–7.87 (m, 14H, ArH), 8.25 (s, 2H, vinyl-H), 8.79 (d, 2H,
Ar-H). MS (EI, 70eV): m/z (%)=526 [M ]. Anal. Calcd for
+
(
2-amino-7,7-dimethyl-5-oxo-5,6,7,8-tetrahydro-4H-chromene-3-
carbonitrile) (5g). Pale yellow crystals (dioxane/ethanol),
C H O : C, 77.56; H, 4.21. Found: C, 77.63; H, 4.16.
3
4 22 6
mp 265–268°C, IR (KBr): ν = 3436, 3350 (NH ), 2195
2,2'-(((Butane-1,4-diylbis(oxy))bis(2,1-phenylene))bis
(methanylylidene))-bis(1H-indene-1,3(2H)-dione) (14b).
Yellow crystals (dioxane) (Method A, 85%, Method B,
2
ꢀ
1 1
(
CN), 1682 (CO) cm , H-NMR (300 MHz, DMSO-d ):
6
δ= 0.95 (s, 6H, 2CH ), 1.08 (s, 6H, 2CH ), 2.04–2.42
3
3
ꢀ
1
(
m, 4H, H8), 2.50 (s, 4H, H6), 4.24–4.35 (m, 4H,
88%), Mp 202–205°C. IR (KBr): ν = 1666 (CO) cm .
1
2
OCH ), 4.50 (s, 2H, pyran H-4), 6.78–7.22 (m, 10H,
H-NMR (300MHz, DMSO-d ): δ= 2.05 (s, 4H, 2CH ),
2
6 2
13
ArH + 2NH2). C-NMR (75 MHz, DMSO-d6): δ = 26.6,
4.27 (s, 4H, 2OCH ) 6.99–7.96 (m, 14H, ArH), 8.27 (s,
2
2
1
8.3, 31.2, 31.7, 50.1, 57.1, 66.3, 111.7, 112.7, 119.9,
2H, vinyl-H), 8.75 (d, 2H, Ar-H). MS (EI, 70eV): m/z
+
20.7, 127.9, 129.2, 131.9, 156.1, 159.0, 162.8, 195.6.
(%) = 554 [M ]. Anal. Calcd for C H O : C, 77.97; H,
36 26 6
+
MS (EI, 70eV): m/z (%)=804 [M ]. Anal. Calcd for
4.73. Found: C, 77.67; H, 4.78.
C H Br N O : C, 56.73; H, 4.51; Br, 19.86; N, 6.96.
Found: C, 56.67; H, 4.59; Br, 19.78; N, 6.78.
2,2'-(((Butane-1,4-diylbis(oxy))bis(4,1-phenylene))bis(methanyly-
lidene))bis(1H-indene-1,3(2H)-dione) (14c).
38
36
2 4 6
Yellow crystals
(dioxane) (Method A, 84%, Method B, 87%), Mp > 300°C.
4
,4'-((Butane-1,4-diylbis(oxy))bis(5-bromo-2,1-phenylene))bis
ꢀ
1
1
(
2-amino-7,7-dimethyl-5-oxo-5,6,7,8-tetrahydro-4H-chromene-3-
IR (KBr): ν = 1676 (CO) cm
.
H-NMR (300 MHz,
carbonitrile) (5h). Pale yellow crystals (dioxane / ethanol),
DMSO-d ): δ= 1.95 (s, 4H, 2CH ), 4.06 (s, 4H, 2OCH )
6
2
2
mp 238–240°C, IR (KBr): ν = 3428, 3316 (NH ), 2188
6.74–8.62 (m, 18H, ArH + vinyl-H). MS (EI, 70eV): m/z
2
ꢀ
1
1
+
(
CN), 1682 (CO) cm
,
H-NMR (300 MHz,
(%) = 554 [M ]. Anal. Calcd for C H O : C, 77.97;
36 26 6
DMSO-d ): δ = 0.93 (s, 6H, 2CH ), 1.04 (s, 6H, 2CH ),
H, 4.73. Found: C, 77.86; H, 4.82.
6
3
3
1
3
6
.89–2.28 (m, 8H, H8+ 2 CH ), 2.50 (s, 4H, H6),
Biology. Stock solutions of the test compounds were
dissolved in dimethyl sulfoxide (DMSO) (Sigma).
Synthetic compounds were 10-fold serially diluted with
Dulbecco's Modified Eagle's Medium. The cytotoxic
effect of each compounds was tested individually in
Madin-Darby Canine kidney (MDCK) cells using the
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
(MTT, Lonza, Wokingham, UK) with minor modification
as previously described.[62] Briefly, MDCK cells were
seeded in 96 well culture plates and incubated for 24h at
2
.96–4.02 (m, 4H, 2OCH ), 4.45 (s, 2H, pyran H-4),
2
1
3
.94–7.35 (m, 10H, ArH + 2NH2). C-NMR (75 MHz,
DMSO-d6): δ= 25.6, 26.2, 28.5, 31.3, 31.7, 49.9, 56.5,
6
1
=
H, 4.84; Br, 19.19; N, 6.73. Found: C, 57.83; H, 4.72;
Br, 19.28; N, 6.78.
General Procedure for the Synthesis of Compounds 14a–c.
Method A: To a mixture of bisaldehydes 1a–c (1mmol),
malononitrile (2.2mmol) and indanedione 11 (2.2mmol)
in absolute ethanol (15mL) was added piperidine (0.2mL)
and the mixture was heated at reflux for 2 h. The crude
solid was isolated and recrystallized from the proper
solvent.
Method B: To a solution of bisaldehydes 1a–c (1mmol) in
absolute ethanol (15 mL), indanedione 11 (2.2 mmol) and
piperidine (0.2 mL) was added and the mixture was
heated at reflux for 1 h. The crude solid was isolated and
recrystallized from the proper solvent.
8.0, 111.5, 114.5, 119.6, 120.0, 130.3, 131.5, 132.7,
55.8, 158.9, 162.2, 195.7. MS (EI, 70 eV): m/z (%)
+
832 [M ], Anal. Calcd for C H Br rN O : C, 57.70;
4
0
40
2
4 6
37°C in 5% CO . After 24h post culture, the cells were
2
treated with desired concentrations of the tested
compounds in triplicates. After further 24h, the medium
was aspirated, and cell monolayer was washed with
phosphate buffer saline three times then MTT solution
(20 μl of 5 mg/ml stock solution) was added to each well
and incubated at 37°C for 4 h. A volume of 200 μl of
DMSO was added to each well to dissolve formazan
crystals. Absorbance of formazan solutions was measured
at λmax 540nm. The percentage of cytotoxicity of each
compound was determined with the following equation.
ðAbsorbance of cell without treatment - Absorbance of cell with treatmentÞ
%
Cytotoxicity ¼
ꢁ100
Absorbance of cell without treatment
2
,2'-(((Ethane-1,2-diylbis(oxy))bis(2,1-phenylene))bis
The plot of % cytotoxicity versus concentration of tested
compounds was used to calculate the concentration, which
exhibited 50% cytotoxicity (TC50).
(
methanylylidene))-bis(1H-indene-1,3(2H)-dione) (14a).
Yellow crystals (dioxane) (Method A, 88%, Method B,
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

A. M. Abdella, Y. Moatasim, M. A. Ali, A. H. M. Elwahy, and I. A. Abdelhamid
Vol 000
Plaque reduction assay.
compounds were carried out using plaque reduction assay
as previously described [63]. Briefly, MDCK cells were
seeded in six well culture plates (10 cells/ml) and
Antiviral activities of tested
[13] Kemnitzer, W.; Drewe, J.; Jiang, S.; Zhang, H.; Zhao, J.;
Crogan-Grundy, C.; Xu, L.; Lamothe, S.; Gourdeau, H.; Denis, R.; Tseng,
B.; Kasibhatla, S.; Cai, S. X. J Med Chem 2007, 50, 2858.
[14] Kemnitzer, W.; Drewe, J.; Jiang, S.; Zhang, H.; Crogan-
5
Grundy, C.; Labreque, D.; Bubenick, M.; Attardo, G.; Denis, R.;
Lamothe, S.; Gourdeau, H.; Tseng, B.; Kasibhatla, S.; Cai, S. X. J Med
Chem 2008, 51, 417.
incubated for 24 h at 37°C in 5% CO . Previously titrated
2
A/duck/Egypt/Q5569D/2012(H5N1) virus was diluted to
optimal virus dilution that gave countable plaques and
mixed with the safe concentration of each tested
compounds then incubated for 1 h at 37°C before being
added to the cells. Growth medium was removed from
the 6-well cell culture plates, and virus-compound
mixtures were inoculated in duplicates, After 1h contact
time for virus adsorption, 3 ml of Dulbecco's Modified
Eagle's Medium supplemented with 2% agarose, 1%
antibiotic antimycotic mixture, and 4% bovine serum
albumin (Sigma) was added onto the cell monolayer,
plates were left to solidify and incubated at 37°C till
formation of viral plaques (3days). Formalin (10%) was
added to each well for 1 h then over layer was removed.
Fixed cells were stained with 0.1% crystal violet in
distilled water. Control untreated virus was included in
each plate. Finally, plaques were counted and percentage
of reduction in virus count was calculated as follows:
[
15] Albiston, A. L.; Diwakarla, S.; Fernando, R. N.; Mountford, S. J.;
Yeatman, H. R.; Morgan, B.; Pham, V.; Holien, J. K.; Parker, M. W.;
Thompson, P. E.; Chai, S. Y. Br J Pharmacol 2011, 164, 37.
[
16] Martínez-Grau, A.; Marco, J. Bioorg Med Chem Lett 1997, 7,
165.
[17] Conti, C.; Desideri, N. Bioorg Med Chem 2009, 17, 3720.
18] Conti, C.; Proietti Monaco, L.; Desideri, N. Bioorg Med Chem
014, 22, 1201.
19] Sangani, C. B.; Shah, N. M.; Patel, M. P.; Patel, R. G. J Ser-
3
2
[
[
bian Chem Soc 2012, 77, 1165.
[20] Thareja, S.; Verma, A.; Kalra, A.; Gosain, S.; Rewatkar, P. V.;
Kokil, G. R. Acta Pol 2010, 67, 423.
[
21] Thomas, N.; Zachariah, S. Asian J Pharm Clin Res 2013,
, 11.
[22] Skommer, J.; Wlodkowic, D.; Mättö, M.; Eray, M.; Pelkonen,
J. Leuk Res 2006, 30, 322.
23] Kemnitzer, W.; Kasibhatla, S.; Jiang, S.; Zhang, H.; Zhao, J.;
6
[
Jia, S.; Xu, L.; Crogan-Grundy, C.; Denis, R.; Barriault, N.; Vaillancourt,
L.; Charron, S.; Dodd, J.; Attardo, G.; Labrecque, D.; Lamothe, S.;
Gourdeau, H.; Tseng, B.; Drewe, J.; Cai, S. X. Bioorg Med Chem Lett
2
005, 15, 4745.
24] El-Bayouki, K.; Aly, M.; Mohamed, Y. World J Chem 2009,
4, 161.
[
[25] Thurston, D. E.; Bose, D. S.; Thompson, A. S.; Howard, P. W.;
Leoni, A.; Croker, S. J.; Jenkins, T. C.; Neidle, S.; Hartley, J. A.;
Hurley, L. H. J Org Chem 1996, 61, 8141.
viral count ðuntreatedÞ - viral count ðtreatedÞ
%
inhibition ¼
ꢁ100
[26] Shaker, R. M. Phosphorus Sulfur Silicon Relat Elem 1999,
viral count ðuntreatedÞ
149, 7.
[27] Dolle, R. E. J Comb Chem 2005, 7, 739.
Acknowledgments.
acknowledge the Alexander von Humboldt Foundation for a
research fellowship.
A.H.M.E.
and
I.A.A.
gratefully
[28] Dolle, R. E. J Comb Chem 2004, 6, 623.
[29] Eccles, R. Lancet Infect Dis 2005, 5, 718.
[30] Mould, J. A.; Drury, J. E.; Frings, S. M.; Kaupp, U. B.;
Pekosz, A.; Lamb, R. A.; Pinto, L. H. J Biol Chem 2000, 275, 31038.
[31] McKimm-Breschkin, J.; Trivedi, T.; Hampson, A.; Hay, A.;
Klimov, A.; Tashiro, M.; Hayden, F.; Zambon, M. Antimicrob Agents
Chemother 2003, 47, 2264.
REFERENCES AND NOTES
[
32] Nguyen, H. T.; Fry, A. M.; Gubareva, L. V. Antivir Ther 2012,
17, 159.
[33] Piñón, A.; Acosta, B.; Valdés, O.; Pérez, A.; Muné, M.;
Arencibia, A.; Savón, C.; Goyenechea, A.; Oropesa, S.; González, G.;
González, G.; Hernández, B. Int J Antimicrob Agents 2013, 42, 97.
[34] Ghozlan, S. A. S.; Abdelmoniem, A. M.; Butenschön, H.;
Abdelhamid, I. A. Tetrahedron 2015, 71, 1413.
[35] Ghozlan, S. A. S.; Mohamed, M. H.; Abdelmoniem, A. M.;
Abdelhamid, I. A. Arkivoc 2009, (x), 302.
[36] Abdelhamid, I. A.; Darwish, E. S.; Nasra, M. A.; Abdel-Gallil,
F. M.; Fleita, D. H. Synthesis (Stuttg) 2010, 1107.
[
1] Rostamnia, S.; Nuri, A.; Xin, H.; Pourjavadi, A.; Hosseini, S. H.
Tetrahedron Lett 2013, 54, 3344.
2] Nemouchi, S.; Boulcina, R.; Carboni, B.; Debache, A.
Comptes Rendus Chim 2012, 15, 394.
3] Borhade, A. V.; Uphade, B. K.; Tope, D. R. J Chem Sci 2013,
25, 583.
[
[
1
2
2
[4] Undale, K.; Park, Y.; Park, K.; Dagade, D.; Pore, D. Synlett
011, 0791.
[
5] Mekheimer, R. A.; Sadek, K. U. Chinese Chem Lett 2009,
0, 271.
[
6] Albadi, J.; Mansournezhad, A.; Darvishi-Paduk, M. Chinese
Chem Lett 2013, 24, 208.
7] Abdelaziz, E. H.; Elnagdi, M. H.; Elagamey, A. A.; El-taweel,
F. M. A. Heterocycles 1987, 26, 903.
8] Abdel-Galil, F. M.; Riad, B. Y.; Sherif, S. M.; Elnagdi, M. H.
Chem Lett 1982, 1123.
[37] Ghozlan, S. A. S.; Mohamed, M. F.; Ahmed, A. G.; Shouman,
S. A.; Attia, Y. M.; Abdelhamid, I. A. Arch. Pharm. (Weinheim) 2015,
3
48, 113.
[38] Abdelhamid, I. A. Synlett 2008, 10, 625.
[39] Abdelhamid, I. A.; Mohamed, M. H.; Abdelmoniem, A. M.;
[
[
Ghozlan, S. A. S. Tetrahedron 2009, 65, 10069.
[40] Ghozlan, S. A. S.; Abdelhamid, I. A.; Hassaneen, H. M.;
Elnagdi, M. H. J Heterocyclic Chem 2007, 44, 105.
[41] Ghozlan, S. A. S.; Abdelhamid, I. A.; Elnagdi, M. H. Arkivoc
2006 147.
[42] Ghozlan, S. A. S.; Ahmed, A. G.; Abdelhamid, I. A.
J Heterocyclic Chem 2015. DOI:10.1002/jhet.2341.
[43] Elwahy, A. H. M. J Chem Res (S) 1999, 602.
[44] Elwahy, A. H. M.; Ahmed, M. M.; El-sadek, M. J Chem Res
2001, 2001, 175.
[9] Al-Matar, H. M.; Khalil, K. D.; Herbert, M.; Kolshorn, H.;
Elnagdi, M. H. Arkivoc 2008, 2008, 288.
[
10] Kumar, B. S.; Srinivasulu, N.; Udupi, R. H.; Rajitha, B.;
Reddy, Y. T.; Reddy, P. N.; Kumar, P. S. Russ J Org Chem 2006, 42, 1813.
11] Gourdeau, H.; Leblond, L.; Hamelin, B.; Desputeau, C.; Dong,
K.; Kianicka, I.; Custeau, D.; Boudreau, C.; Geerts, L.; Cai, S.-X.; Drewe,
J.; Labrecque, D.; Kasibhatla, S.; Tseng, B. Mol Cancer Ther 2004, 3, 1375.
[
[
12] Mohr, S. J.; Chirigos, M. A.; Fuhrman, F. S.; Pryor, J. W.
Cancer Res 1975, 35, 3750.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2016
Anti-influenza Virus Activity of bis(4H-chromene-3-carbonitrile) Derivatives
[45] Elwahy, A. H. M.; Abbas, A. A. Synth Commun 2000, 30,
[55] Bernasconi, C. F.; Kanavarioti, A.; Killion, R. B. J Am Chem
2
903.
Soc 1985, 107, 3612.
[
46] Elwahy, A. H. M.; Abbas, A. A. Kassab, R. M. Synthesis
Stuttg) 2002, 260.
47] Abbas, A. A.; Elwahy, A. H. M.; Ahmed, A. A. M. J Heterocy-
clic Chem 2005, 42, 93.
48] Elwahy, A. H. M.; Masaret, G. S. J Heterocyclic Chem 2007,
4, 1475.
49] Muathen, H. A.; Aloweiny, N. A. M.; Elwahy, A. H. M.
J Heterocyclic Chem 2009, 46, 656.
50] Shaaban, M. R.; Elwahy, A. H. M. J Heterocyclic Chem 2012,
9, 640.
[56] Bernasconi, C. F.; Fox, J. P.; Kanavarioti, A.; Panda, M. J Am
Chem Soc 1986, 108, 2372.
(
[
[57] Sofan, M. A.; El-Taweel, F. M.; Elagamey, A. G. A.; Elnagdi,
M. H. Liebigs Ann Der Chemie 1989, 1989, 935.
[58] Girgis, N. S.; Elgemeie, G. E. H.; Nawar, G. A. M.; Elnagdi,
M. H. Liebigs Ann Der Chemie 1983, 1983, 1468.
[59] Elgemeie, G. E. H.; Riad, B. Y.; Nawwar, G. A.; Elgamal, S.
Arch. Pharm. (Weinheim) 1987, 320, 223.
[
4
[
[
[60] Sharanin, Y. A.; Goncharenko, M. P.; Litvinov, V. P. Russ
Chem Rev 1998, 67, 393.
4
[
51] Elwahy, A. H. M. Tetrahedron 2000, 56, 897.
[61] Abbas, A. A.; Elneairy, M. A. A.; Mabkhot, Y. N. J Chem Res
2001, 124.
[52] Elwahy, A. H. M.; Abbas, A. A. Tetrahedron 2000, 56,
8
85.
[
[62] Rashad, A. E.; Shamroukh, A. H.; Abdel-Megeid, R. E.;
Mostafa, A.; El-Shesheny, R.; Kandeil, A.; Ali, M. A.; Banert, K. Eur J
Med Chem 2010, 45, 5251.
[63] Hayden, F. G.; Cote, K. M.; Douglas, R. G. Antimicrob
Agents Chemother 1980, 17, 865.
53] Qareaghaj, O. H.; Mashkouri, S.; Naimi-Jamal, M. R.;
Kaupp, G. RSC Adv 2014, 4, 48191.
54] Abdel-Rahman, N. M.; El-Kateb, A. A.; Mady, M. F. Synth
Commun 2007, 37, 3961.
[
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet