January 2017
Synthesis and Evaluation of Pyrido[2,3-d]pyrimidine and 1,8Naphthyridine Derivatives as Potential Antitumor Agents
295
Mohamed S. Behaloa* and Giuseppe Meleb
a
Chemistry Department, Faculty of Science, Benha University, 13518 Benha, Egypt
b
Department of Engineering for Innovation, University of Salento, Lecce, Italy
*E-mail: [email protected]
Received September 7, 2015
DOI 10.1002/jhet.2581
Published online 11 January 2016 in Wiley Online Library (wileyonlinelibrary.com).
New series of pyrido[2,3-d]pyrimidine and 1,8-naphthyridine derivatives were synthesized from 2-amino-6(phenoxathiin-2-yl)-4-(thiophene-2-yl) nicotinonitrile as starting material, and their structures were characterized
on the basis of the spectral data. Most of the synthesized compounds were evaluated for their cytotoxic
activity against two cancer cell lines, namely, breast cancer Michigan Cancer Foundation-7 (MCF-7)
and prostate cancer human prostatic carcinoma cell line (PC-3) using MTT assay. Some of these compounds showed potent cytotoxic effect concluded from their IC50 values.
J. Heterocyclic Chem., 54, 295 (2017).
INTRODUCTION
Pyridopyrimidine and its derivatives are an important
group of heterocyclic compounds, which have been subject
to extensive study in the past years because of a variety of
chemical and biological significance. The importance of
pyridopyrimidines as biologically active compounds includes their use as antitumor [1–5], antimicrobial [6–8],
anti-inflammatory [9,10], antimalarial [11], antifolate [12],
anticonvulsant [13], analgesic [14], and antioxidant [15]. In
addition, pyridopyrimidine-2-thiones displayed potent activity against Mycobacterium tuberculosis H37Rv [16].
Pipemidic acid, 8-ethyl-5,8-dihydro-5-oxo-2-(1-piperazinyl)pyrido[2,3-d]pyrimidine-6-carboxylic acid, is potent antibacterial agent (Fig. 1) [17]. In addition, 1,8-naphthyridine
derivatives have been described as potential analgesic [18],
antitumor [19,20], antiplatlet [21], and antihypertensive
agents [22].
Nalidixic acid, 1-ethyl-7-methyl-4-oxo-1,4-dihydro-1,8naphthyridine-3-carboxylic acid, is the first of the synthetic
naphthyridine antibiotics [23]. It has historically been used
for treating urinary tract infections caused by Escherichia
coli, Proteus, and Klebsiella
On the basis of the earlier-mentioned facts and in
continuation of our ongoing interest in the discovery
of new biologically active heterocycles [24–28], the
present synthetic protocol involves synthesis of new
series of pyridopyrimidine and 1,8-naphthyridine derivatives from 2-amino-6-(phenoxathiin-2-yl)-4-(thiophene-2-yl) nicotinonitrile as starting material and
evaluation of their cytotoxic activity against two cancer cell lines namely breast cancer Michigan Cancer
Foundation (MCF)-7 and prostate cancer PC-3 using
MTT assay.
RESULTS AND DISCUSSION
Chalcone, E-1-(phenoxathiin-2-yl)-3-(thiophene-2-yl)prop2-en-1-one (1) (prepared from the treatment of thiopene-2carbaldhayde with 2-acetylphenoxathiin), reacted with
malononitrile in the presence of ammonium acetate to afford
© 2016 HeteroCorporation
296
M. S. Behalo and G. Mele
Figure 1. Molecular structure of Pipemidic and Nalidixic acid.
the required starting material 2-amino-6-(phenoxathiin-2yl)-4-(thiophene-2-yl) nicotinonitrile (2) in good yield
(Schemes 1 and 2).
Chalcone 1 can have two stereoisomeric structures, Z
and E forms (Fig. 2), but on the basis of the 1H NMR spectrum that showed two doublet signals for the two olefinic
protons at 7.31 and 7.73 ppm with the coupling constant
value J = 15.7 Hz, it seems to exist predominately in the E
form [29]. On the other hand, the structure of compound
2 was established on the basis of its spectral data. The IR
spectrum showed strong absorption bands of amino and nitrile groups at 3381, 3156, and 2214 cm1, respectively.
Also, 1H NMR spectrum revealed a signal at 5.62 ppm.
for the NH2 group.
The presence of amino and nitrile group at 1,2-position
to each other on the pyridine ring gives compound 2 significant importance as a reactive key precursor for the construction of heterocyclic systems of expected biological
activity. Thus, fusion of compound 2 with urea or thiourea
afforded pyrido[2,3-d]pyrimidine 3a,b, respectively [24].
The structures of products 3a,b were established on the basis of disappearance of a band characteristic to nitrile group
in IR spectrum and their mass spectra that showed molecular ion peaks m/z at 442 (M+, 77%) and 458 (M+, 82%),
respectively.
On the other hand, treatment of compound 2 with urea in
ethanol containing sodium ethoxide gave 1-(3-cyano-6(phenoxathiin-2-yl)-4-(thiophen-2-yl)pyridin-2-yl)urea
(4), which upon heating above its melting point, furnished
pyridopyrimidine 3a. The reaction probably takes place
through the elimination of ammonia followed by intramolecular cyclization at the cyano group.
On the other hand, cyclocondensation of 2 with phenyl
isothiocaynate furnished the corresponding pyridopyrimidinethione derivative 6. The reaction probably takes
Vol 54
place via formation of thiourea intermediate 5 followed
by cyclization at the adjacent C≡N group [24].
Aminopyridopyrimidine 7 can be synthesized through
the reaction of compound 2 with formamide. Also, heating
of 2 with formic acid under reflux afforded pyrido[2,3-d]
pyimidine 8 (Scheme 3), the IR spectrum displayed a lack
of absorption bands of both C≡N and NH2 groups and the
presence of CO and NH absorption bands (Scheme 4).
On the other hand, treatment of compound 2 with carbon
disulfide in dimethylformamide afforded the reaction product that could be formulated as 7-(phenoxathiin-2-yl)-5(thiophen-2-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dithione
(9); the reaction is assumed to take place through Dimroth
rearrangement [30].
This investigation was extended to use compound 2 as
a reactive substrate for the synthesis of a number of
biologically active 1,8-naphthyridines. Thus, treatment of
compound 2 with active methylene compounds, namely,
ethyl cyanoacetate and ethyl acetoacetate under reflux in
dimethylformamide in the presence of catalytic amount of
piperidine afforded 1,8-naphthridines 11a,b, respectively.
The reaction takes place through formation of intermediate
10 formed from elimination of ethanol followed by cyclization at cyano group and tautomerization [31]. On the
other hand, heating of compound 2 with malononitrile in
dimethylformamide in the presence of catalytic amount of
piperidine furnished 2,4-diamino-7-(phenoxathiin-2-yl)-5(thiophen-2-yl)-1,8-naphthridin-3-carbonitrile (12).
However, cyclocondensation of compound 2 with
benzylidenemalononitrile in ethanol containing few
drops of piperidine gave 1,8-naphthridine 13.
CYTOTOXICITY ASSAY
Materials and methods. Cell line. Two human tumor
cell lines are, namely, mammary gland (breast) MCF-7
and human (prostate) cancer cell line PC-3. The cell
lines were obtained from American Type Culture
Collection (ATCC, Manassas, VA) via Holding company
for biological products and vaccines (VACSERA, Cairo,
Egypt).
Scheme 1. Synthesis of 2-amino-6-(phenoxathiin-2-yl)-4-(thiophene-2-yl) nicotinonitrile (2). [Color figure can be viewed in the online issue, which is
available at wileyonlinelibrary.com.]
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet
January 2017
Antitumors
297
Scheme 2. Synthesis of pyrido[2,3-d]pyrimidines 3–6. [Color figure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]
Chemical reagents. The reagents RPMI-1640 medium,
MTT, and dimethyl sulfoxide (DMSO) and 5-fluorouracil
were purchased by Sigma Co. (St. Louis, USA) and the
fetal bovine serum by GIBCO (UK).
MTT assay.
The different cell lines mentioned earlier
were used to determine the inhibitory effects of compounds
on cell growth using the MTT assay. This colorimetric
assay is based on the conversion of the yellow tetrazolium
bromide (MTT) to a purple formazan derivative by
mitochondrial succinate dehydrogenase in viable cells [32].
The cells were cultured in RPMI-1640 medium with 10%
fetal bovine serum. Antibiotics added were 100-units/mL
penicillin and 100-μg/mL streptomycin at 37°C in a 5%
CO2 incubator. The cells were seeded in a 96-well plate at
a density of 1.0 × 104 cells/well at 37°C for 48 h under 5%
CO2. After incubation, the cells were treated with different
concentration of compounds and incubated for 24 h. After
24 h of drug treatment, 20 μL of MTT solution at 5 mg/mL
was added and incubated for 4 h. DMSO in volume of
100 μL is added into each well to dissolve the purple
formazan formed. The colorimetric assay is measured and
recorded at absorbance of 570 nm using a plate reader
(EXL 800). The relative cell viability in percentage was
calculated as (A570 of treated samples/A570 of untreated
sample) × 100.
Antitumor activity. Treatment of cell lines MCF-7 and
PC-3 with the samples showed different cytotoxic effect as
shown in Table 1.
It was observed that pyrido[2,3-d]pyrimidinethione
3b, pyrido[2,3-d] pyrimidinedithione 9 and 1,8-naphthridincarbonitrile 12 showed the most potent cytotoxic
effect against prostate cancer cell line PC-3 concluded
from their IC50 values 9.47, 10.34, and 8.13 μg/mL respectively, but both of compounds 8 and 13 showed no
cytotoxicity. On the other hand, only naphthyridine derivative 12 showed very strong cytotoxic effect against
breast cancer MCF-7.
CONCLUSION
In summary, we have synthesized novel derivatives of
pyrido[2,3-d]pyrimidine and 1,8-naphthyridine from 2Scheme 3. Synthesis of pyrido[2,3-d]pyrimidines 7–9. [Color figure can be
viewed in the online issue, which is available at wileyonlinelibrary.com.]
Figure 2. Z and E forms of chalcone 1.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet
298
M. S. Behalo and G. Mele
Scheme 4. Synthesis of 1,8-naphthyridines 11–13. [Color figure can be
viewed in the online issue, which is available at wileyonlinelibrary.com.]
Vol 54
CA, USA). The solvent used for NMR analysis was
DMSO-d6, unless stated otherwise. Mass spectra were
obtained using a Shimadzu GCMS-QP 1000 EX mass
spectrometer (Kyoto, Japan).
E-1-(Phenoxathiin-2-yl)-3-(thiophene-2-yl)prop-2-en-1one (1).
A mixture of 2-acetylphenoxathiin (0.01 M),
amino-6-(phenoxathiin-2-yl)-4-(thiophene-2-yl) nicotinonitrile,
and their structures were characterized on the basis of the
spectral data and elemental analyses. Most of the synthesized
compounds were evaluated for their cytotoxic activity against
two cancer cell lines, and it was observed that pyrido[2,3-d]
pyrimidinethione 3b, pyrido[2,3-d] pyrimidinedithione 9
and 1,8-naphthridincarbonitrile 12 showed the most potent
cytotoxic effect against prostate cancer cell line PC concluded
from their IC50 values.
EXPERIMENTAL
Melting points of the prepared products are uncorrected.
All reactions were monitored by thin layer chromatography
carried out on 0.2-mm silica gel 60 F254 plates (Merck,
Kenilworth, NJ, USA). IR spectra in KBr were recorded
using a Perkin-Elmer 298 spectrophotometer (Waltham,
MA, USA). 1H and 13CNMR spectra were obtained using
a Varian Gemini 300 and 50-MHz instrument (Palo Alto,
thiophene-2-carbaldehyde (0.01 M), and sodium hydroxide
(1 g in 10-mL water) in ethanol (30 mL) was stirred at RT
for about 3 h. The formed solid was filtered, washed, dried,
and crystallized from ethanol to give yellow crystals of
chalcones 1. Yield, 77%; mp 180–182°C. IR (KBr, ν
cm1): 1681 (C¼O), 1598 (C¼C); 1H NMR (DMSO-d6):
δ ppm: 7.31 (d, 1H, α-CH olefinic, J = 15.7 Hz), 7.73 (d,
1H, β-CH olefinic, J = 15.7 Hz), 7.28–8.07 (m, 10H, 13C
NMR: δ 113.8, 116.7, 117.8, 118.2, 121.3, 125.2, 127.1,
128.2, 128.5, 129.2, 129.8, 130.6, 131.3, 133.5, 140.2,
149.4, 152.2, 183.6, MS: m/z 336 (M+). Anal. Calcd for
C19H12O2S2 (336.03): C, 67.83; H, 3.60 Found: C, 67.75;
H, 3.55.
2-Amino-6-(phenoxathiin-2-yl)-4-(thiophene-2-yl) nicotinonitrile
(2). A mixture of chalcone (0.01 M) 1 and malononitrile
(0.01 M) in EtOH (20 mL) containing ammonium acetate
(0.02 M) was heated under reflux for 6 h. The reaction
mixture was concentrated, cooled, and filtered, and the
precipitated solid was crystallized from ethanol to give
2. Yield, 72%; mp 203–205°C. IR: KBr, ν cm1), 2214
(C≡N), 3381, 3156 cm1 (NH2). 1H NMR (DMSO-d6):
δ 5.62 (s, 2H, NH2, exchangeable), 6.89–8.03 (m, 11H,
ArH). 13C NMR: δ 112.9, 113.5, 116.3, 117.5, 118.9,
119.3, 121.6, 124.3, 125.4, 127.6, 127.9, 128.3, 129.3,
129.8, 130.5, 133.2, 137.6, 148.2, 150.1, 151.5, 159.3,
MS: m/z 399 (M+). Anal. Calcd. for C22H13N3OS2
(399.05): C, 66.14; H, 3.28; N, 10.52. Found: C, 66.05;
H, 3.16; N, 10.43.
Synthesis of 3a,b: General Procedure.
An equimolar
amount of 2 and urea or thiourea (0.01 M) was fused in an oil
bath for 2 h. After cooling, the product was treated with
water, filtered, dried, and crystallized to give 3a,b, respectively.
Table 1
Cytotoxicity of tested compounds on different cell lines.
In vitro cytotoxicity IC50 (μg/mL)
Compounds
5-Fu
3b
6
8
9
11a
12
13
PC-3
MCF-7
4.91 ± 0.38
9.47 ± 0.77
33.42 ± 2.50
>100
10.34 ± 2.55
31.25 ± 1.63
8.13 ± 0.88
>100
4.73 ± 0.47
21.38 ± 1.53
50.06 ± 2.57
87.35 ± 3.72
58.36 ± 2.88
29.58 ± 1.34
7.93 ± 0.36
>100
IC50 (μg/mL): 1–10 (very strong), 11–20 (strong), 21–50 (moderate), 51– 100(weak), and above 100 (non-cytotoxic)
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet
January 2017
Antitumors
4-Amino-7-(phenoxathiin-2-yl)-5-(thiophen-2-yl)pyrido[2,3d]pyrimidin-2(1H)-one 3a. Yield, 82% (EtOH); mp 235–
237°C. IR: KBr, ν cm1), 1670 (CO), 3432–3240 (NH2
and NH); 1H NMR (DMSO-d6): δ 6.81 (s, 2H, NH2,
exchangeable), 7.52–8.02 (m, 11H, Ar H), 8.64 (s, 1H,
NH, exchangeable). 13C NMR: δ 110.6, 113.3, 115,8,
116.2, 117.2, 118.4, 121.2, 124.9, 125.6, 126.2, 127.5,
128.3, 129.1, 129.8, 130.3, 135.2, 142.5, 146.5, 147.2,
150.5, 151.3, 152.4, 154.6; Anal. Calcd for C23H14N4O2S2
(442.06): C, 62.43; H, 3.19; N, 12.66. Found: C, 62.35; H,
3.15; N, 12.55.
4-Amino-7-(phenoxathiin-2-yl)-5-(thiophen-2-yl)pyrido[2,3d]pyrimidin-2(1H)-thione 3b. Yield, 78%; mp 213–215°C.
IR (KBr, ν cm1): 1262 (CS), 3437–3200 cm1 (NH2, NH).
1
H NMR (DMSO-d6): δ 8.45 (s, 2H, NH2, exchangeable),
9.25 (s, 1H, NH), 6.96–7.88 (m, 11H, ArH). MS: m/z: 458
(M+); Anal. Calcd for C23H14N4OS3 (458.03) C, 60.24; H,
3.08; N, 12.22. Found: C, 60.15; H, 3.03; N, 12.16.
1-(3-Cyano-6-(phenoxathiin-2-yl)-4-(thiophen-2-yl)pyridin2-yl)urea (4). A mixture of compound 2 (0.01 M) and urea
(0.01 M) was refluxed in ethanol containing sodium ethoxide
(0.5-g sodium in 30-mL absolute ethanol) for 5 h. After
cooling, the reaction mixture was poured into crushed ice
and HCl. The formed solid precipitate was collected by
filteration, washed with water, and recrystallized from
ethanol to give compound 4. Yield, 75%; mp 196–198°C.
(IR KBr, ν cm1): 1665 (CO), 2214 (C≡N), 3384–
3154 cm1 (NH, NH2). 1H NMR (DMSO-d6): δ 8.72 (s, 2H,
NH2, exchangeable), 12.19 (s, 1H, NH), 7.18–8.19 (m, 11H,
ArH). Anal. Calcd for C23H14N4O2S2 (442.06): C, 62.43; H,
3.19; N, 12.66. Found 62.31; H, 3.12; N, 12.58.
4-Imino-3-phenyl-7-(phenoxathiin-2-yl)-5-(thiophen-2-yl)-3,4dihydropyrido [2,3-d]pyrimidin-2(1H)-thione 6.
A mixture
of 2 (0.01 M) and phenyl isothiocyanate (0.01 M) in
pyridine (15 mL) was refluxed for 5 h. After cooling, the
reaction mixture was poured into cold water containing
HCl. The obtained solid product was filtered, dried, and
crystallized from dioxan. Yield, 77%; mp 203–205°C.
IR (KBr, ν cm1): 1290 (CS) and 3315 (NH); 1H NMR
(DMSO-d6), δ ppm=7.02–8.11 (m, 16H, ArH), 8.23, 8.81
(s, 2H, 2NH exchangeable); MS: m/z: 534 (M+); Anal.
Calcd for C29H18N4OS3 (534.06): C, 65.14; H, 3.39; N,
10.48% Found: C, 65.10; H, 3.33; N, 10.39%.
4-Amino-7-(phenoxathiin-2-yl)-5-(thiophen-2-yl)pyrido[2,3-d]
pyrimidine 7.
A mixture of compound 2 (0.01 M) and
formamide (15 mL) was heated under reflux for 8 h. After
cooling, the solid product was filtered and crystallized from
methanol to give 7. Yield, 70%; mp 238–240°C; IR: (KBr,
ν cm1): 3390, 3340 (NH2), 1620 cm1 (C¼N); 1H NMR
(DMSO-d6), δ ppm = 6.92–8.02 (m, 12H, ArH), 8.32 (s,
2H, NH2 exchangeable); 13C NMR: δ 111.2, 116.5,
117.6, 118.2, 120.3, 122.2, 125.2, 126.8, 127.6, 128.2,
129.3, 129.8, 130.2, 140.2, 142.2, 148.6, 151.3, 152.5,
153.3, 154.3, 155.4, MS: m/z: 426 (M+); Anal. Calcd for
299
C23H14N4OS2 (426.06): C, 64.77; H, 3.31; N, 13.14%.
Found: C, 64.68; H, 3.25; N, 13.06%.
7-(Phenoxathiin-2-yl)-5-(thiophen-2-yl)pyrido[2,3-d]pyrimidin4(3H)-one 8. Compound 2 (0.01 M) was heated with excess
of formic acid under reflux for 4 h. After cooling, the
precipitated solid was filtered, dried, and crystallized from
ethanol and water. Yield, 69%; mp 214–216°C; IR (KBr, ν
cm1): 1608 (C¼N), 1685 (CO) and 3384 cm1 (NH↔OH);
1
H NMR (DMSO-d6), δ ppm = 7.19–7.85 (m, 12H, ArH),
8.52 (s, 1H, NH exchangeable); 13C NMR: δ 114.5, 115.7,
118.3, 118.8, 119.5, 121.3, 124.5, 126.4, 128.3, 128.8,
129.6, 130.7, 131.4, 141.4, 142.5, 147.2, 152.3, 152.8,
153.2, 154.6, 180.5; MS: m/z: 427 (M+); Anal. Calcd for
C23H13N3O2S2 (427.04): C, 64.62; H, 3.07; N, 9.83%.
Found: C, 64.51; H, 3.05; N, 9.75%.
7-(Phenoxathiin-2-yl)-5-(thiophen-2-yl)pyrido[2,3d]pyrimidine-2,4(1H,3H)-dithione 9.
Carbon disulfide
(0.015 M) was added to solution of compound 2 (0.01 M) in
DMF (20 mL). Then, sodium methoxide (10 mL) [prepared
from sodium (0.5 gm) and methanol (10 mL)] was added,
and all were heated under reflux for 10 h. The reaction
mixture was cooled and poured into cold water followed by
addition of NaOH (10 mL) and left overnight. Clear solution
obtained by filtration was acidified by AcOH to give solid
product, which in turn was collected by filtration, dried, and
crystallized from benzene to give 9. Yield, 71%; mp
202–204°C; IR (KBr, ν cm1): 3435–3250 (2NH), 1299–
1256 cm1 (2CS); 1H NMR (DMSO-d6), δ ppm = 7.29–7.95
(m, 11H, ArH), 8.62, 12.65 (2 s, 2H, 2NH) MS: m/z: 475
(M+) 476 (M+1) Anal. Calcd for C23H13N3OS4 (474.99): C,
58.08; H, 2.75; N, 8.83% Found: C, 58.15; H, 3.80; N, 8.79%.
General procedure for synthesis of 11 and 12. A mixture
of compound 2 (0.01 M) and active methylene compounds,
namely, ethyl cyanoacetate, ethyl acetoacetate, or
malononitrile (0.01 M) was heated in dimethylformamide
containing few drops of piperidine under reflux for 3 h;
after cooling, the solid product was collected by filteration
and recrystallized from a proper solvent to give products
11 and 12, respectively.
4-Amino-2-oxo-7-(phenoxathiin-2-yl)-5-(thiophen-2-yl)1,2-dihydro-1,8-naphthridine-3-carbonitrile 11a. Yield, 82%
(EtOH); mp 233–235°C; IR (KBr, ν cm1): 1602 (C¼N),
1671 (CO), 2217 (C≡N) and 3395–3103 cm1 (NH, NH2);
1
H NMR (CDCl3), δ ppm = 5.63 (s, 1H, NH exchangeable),
7.03–8.09 (m, 11H, ArH), 8.95 (s, 2H, NH2 exchangeable);
MS: m/z: 466 (M+); Anal. Calcd for C25H14N4O2S2
(466.06): C, 64.36; H, 3.02; N, 12.01%. Found: C, 64.33;
H, 2.85; N, 11.88%.
3-Acetyl-4-amino-7-(phenoxathiin-2-yl)-5-(thiophen-2yl)-1,8-naphthridin2(1H)-one 11b.
Yield, 86% (EtOH);
mp 242–244°C; IR: (KBr, ν cm1): 1618 (C¼N), 1667
(CO) and 3346, 3224, 3145 cm1 (NH2, NH); 1H NMR
(DMSO-d6), δ ppm = 3.78(s, 3H, CH3), 6.94–7.61 (m, 11H,
ArH), 8.93 (s, 2H, NH2 exchangeable), 13.25 (s, 1H, NH
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet
300
M. S. Behalo and G. Mele
exchangeable); 13C NMR: δ 28.5, 110.2, 111.6, 112.5, 112.8,
117.4, 118.2, 122.3, 124.1, 125.3, 126.3, 127.1, 128.2, 128.8,
129.3, 130.4, 132.5, 146.3, 142.2, 150.6,151.2 152.5,
160.3, 163.2, 186.3, MS: m/z: 483 (M+); Anal. Calcd for
C26H17N3O3S2 (483.07): C, 64.58; H, 3.54; N, 8.69%.
Found: C, 64.48; H, 3.47; N, 8.61%.
2,4-Diamino-7-(phenoxathiin-2-yl)-5-(thiophen-2-yl)-1,8naphthridin-3-carbonitrile 12.
Yield, 65% (EtOH); mp
226–228°C; IR (KBr, ν cm1): 1638 (C¼N), 2215 (C≡N),
3395–3103 cm1 (NH2); 1H NMR (DMSO-d6), δ
ppm = 6.97–8.13 (m, 11H, ArH + pyrimidine H), 8.65, 9.52
(2 s, 4H, 2NH2 exchangeable); MS: m/z: 465 (M+); Anal.
Calcd for C25H15N5OS2 (465.07): C, 64.50; H, 3.25; N,
15.04%. Found: C, 64.52; H, 3.20; N, 14.98%.
4-Amino-2-phenyl-7-(phenoxathiin-2-yl)-5-(thiophen-2-yl)1,8-naphthridin-3-carbonitrile 13. A mixture of compound
2 (0.01 M) and benzylidenemalononitrile (0.01 M) in dioxan
(20 mL) containing sodium (0.5 gm) was refluxed for 3 h.
The reaction mixture was cooled and poured onto crushed
ice and hydrochloric acid. The precipitated solid was
collected by filteration, dried, and recrystallized from
ethanol. Yield, 77%; mp 215–217°C; IR (KBr, ν cm1):
1593 (C¼N), 2210 C≡N and 3328, 3230 cm1 (NH2); 1H
NMR (DMSO-d6), δ ppm = 6.76–7.58 (m, 16H, ArH), 8.83
(s, 2H, NH2 exchangeable); MS: m/z: 526 (M+); Anal.
Calcd for C31H18N4OS2 (526.09): C, 70.70; H, 3.45; N,
10.64%. Found: C, 70.63; H, 3.35; N, 10.58%.
Acknowledgments. The authors wish to thank Benha University
and Ministry of Scientific Research, Egypt for the financial
support. Salento University and Ministero degli Affari EsteriUfficio IV Della D.G.S.P, Italy for the financial support.
REFERENCES
[1] Grivsky, E. M.; Lee, S.; Sigel, C. W.; Duch, D. S.; Nichol, C. A.
J Med Chem 1980, 23, 327.
[2] Saurat, T.; Buron, F.; Rodrigues, N.; Tauzia, M. L.;
Colliandre, L.; Bourg, S.; Bonnet, P.; Guillaumet, G.; Akssira, M.;
Corlu, A.; Guillouzo, C.; Berthier, P.; Rio, P.; Jourdan, M. L.;
Bénédetti, H.; Routier, S. J Med Chem 2014, 57, 613.
[3] Fares, M.; Abou-Seri, S. M.; Abdel-Aziz, H. A.; Abbas, S. E.;
Youssef, M. M.; Eladwy, R. A. Eur J Med Chem 2014, 83, 155.
[4] Moreno, E.; Plano, D.; Lamberto, I.; Font, M.; Encío, I.; Palop,
J. A.; Sanmartín, C. Eur J Med Chem 2012, 47, 283.
[5] Zhang, J. P.; Huang, J.; Liu, C.; Lu, X.; Wu, B.; Zhao, L.; Lu, N.;
Guo, Q.; Li, Z. Chin Chem Lett 2014, 25, 1025.
[6] Ribble, W.; Hill, W. E.; Ochsner, U. A.; Jarvis, T. C.; Guiles,
J. W.; Janjic, N.; Bullard, J. M. Antimicrob Agents Chemother 2010, 54,
4648.
Vol 54
[7] Verma, A. K.; Singh, A. K.; Islam, M. M. Int J Pharm Sci
2014, 6, 341.
[8] Kheder, N. A.; Mabkhot, Y. N.; Farag, A. M. Synth Commun
Acta Cytologica 2008, 38, 3170.
[9] Mohamed, N. R.; Abdelhalim, M. M.; Khadrawy, Y. A.;
Elmegeed, G. A.; Abdel-Salam, O. M. Steroids 2012, 77, 1469.
[10] Nofal, Z. M.; Fahmy, H. H.; Zarea, E. S.; El-Eraky, W. Acta
Pol Pharm 2011, 68(4), 507.
[11] Mane, U. R.; Mohanakrishnan, D.; Sahal, D.; Murumkar, P. R.;
Gridihar, R.; Yaday, M. R. Eu.r J Med Chem 2014, 79, 422.
[12] Rosowsky, A.; Mota, C. E.; Queener, S. F. J Heterocycl Chem
1995, 32, 335–340.
[13] Deyanov, A. B.; Niyazov, R. K.; Nazmetdivov, F. Y.;
Syropyatov, B. Y.; Kolla, V. E.; Konshin, M. E. J Pharm Chem 1991,
25, 248.
[14] Dinakaran, S.; Keloth, K.; Bomma, V. Der Pharma Chemica
Acta cytologica 2012, 4, 255.
[15] Maheswaran, N.; Saleshiera, M. F.; Mahalakshmia, K.;
Sureshkannan, V.; Parthiban, N.; Reddy, A. Int J Chem Sci 2012, 10, 43.
[16] Rajesh, S. M.; Kumar, R. S.; Libertsen, L. A.; Perumal, S.;
Yogeeswari, P.; Sriram, D. Bioorg Med Chem Lett 2011, 21, 3012.
[17] Shimizu, M.; Takase, Y.; Nakamura, S.; Katae, H.; Minami, A.;
Nakata, K.; Inoue, S.; Ishiyama, M.; Kubo, Y. Antimicrob Agents
Chemother 1975, 8, 132.
[18] Di Braccio, M.; Grossi, G.; Alfei, S.; Ballabeni, V.; Tognolini, M.;
Flammini, L.; Giorgio, C.; Bertoni, S.; Barocelli, E. Eur J Med Chem
2014, 86, 394.
[19] Kaila, N.; Green, N.; Li, H.; Hu, Y.; Janz, K.; Gavrin, L. K.;
Thomason, J.; Tam, S.; Powell, D.; Cuozzo, J.; Hall, J. P.; Telliez, J.;
Hsu, S.; Nutter, C. K.; Wang, Q.; Lin, L. Bioorg Med Chem 2007, 15,
6425.
[20] Zhang, S. X.; Bastow, K. F.; Tachibana, Y.; Kuo, S. C.;
Hamel, E.; Mauger, A.; Narayanan, V. L.; Lee, K. H. J Med Chem
1999, 42, 4081.
[21] Ferrarinia, P. L.; Badawnehb, M.; Franconic, F.; Maneraa, C.;
Micelid, M.; Moria, C.; Saccomannia, G. Il Farmaco 2001, 56, 311.
[22] Ferrarinia, P. L.; Moria, C.; Calderoneb, V.; Calzolaria, L.;
Nierib, P.; Saccomannia, G.; Martinottib, E. Eur J Med Chem 1999, 34,
505.
[23] Lesher, G. Y.; Froelich, E. J.; Gruett, M. D.; Bailey, J. H.;
Brundage, R. P. J Med Pharm Chem 1962, 91, 1063.
[24] Behalo, M. S. Phos, Sulfur and Silicon Related Elements 2009,
184, 206.
[25] Attanasi, O. A.; Crescentini, L. D.; Favi, G.; Filippone, P.;
Giorgi, G.; Mantellini, F.; Moscatelli, G.; Behalo, M. S. Organic Letters
2009, 11, 2265.
[26] Attanasi, O. A.; Behalo, M. S.; Favi, G.; Lomonaco, D.;
Mazzetto, S. E.; Mele, G.; Vasapollo, G. Current Org Chem 2012, 16,
2613.
[27] Wasfy, A. F.; Behalo, M. S.; Aly, A. A.; Sobhi, N. M. Chem
Process Eng Res 2013, 10, 20.
[28] Behalo, M. S. J Sulf Chem 2010, 31, 287.
[29] Dudely, H.; William, I. F. Spectroscopic methods in organic
chemistry, 3rd ed.; Maidenhead: Berkshire-England, 1980.
[30] Oganisyan, A. S.; Noravyan, A. S.; Grigoryan, M. Z. Chem
Heterocyclic Compounds 2001, 37, 763.
[31] El-Gaby, M. S.; Abedel-Gawad, S. M.; Ghorab, M. M.; Heiba,
H. I.; Aly, H. M. Phosp, Sulfur, Silicon 2006, 181, 279.
[32] Mauceri, H. J.; Hanna, N. N.; Beckett, M. A.; Gorski, D. H.;
Staba, M. J.; Stellato, K. A.; Bigelow, K.; Heimann, R.; Gately, S.;
Dhanabal, M.; Soff, G. A.; Sukhatme, V. P.; Kufe, D. W.; Weichselbaum,
R. R. Nature 1998, 394, 287.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet