Synthesis of Novel 4 H-Chromenes Containing a Pyrimidine-2-Thione Function in the Presence of Fe3O4 Magnetic Nanoparticles and Study of Their Antioxidant Activity

Article abstract of DOI:10.1080/10426507.2014.884094

The aim of the present work was to search for identification of novel reactive oxygen species (ROS) scavengers by testing new fused chromenopyrimidinethiones, which was synthesized using Fe₃O₄ nanoparticles. The new compounds were also tested for their cytotoxic activity. The obtained results showed that the incorporated pyrimidinethione moiety significantly increase antioxidant activity. In conclusion, the study of the pharmacological properties of the new chromenopyrimidinethiones allowed establishing new structure-activity relationships for splitting antioxidant and cytotoxic activity of these compounds.

Full text of DOI:10.1080/10426507.2014.884094

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Synthesis of Novel 4H-Chromenes
Containing a Pyrimidine-2-Thione
Function in the Presence of Fe3O4
Magnetic Nanoparticles and Study of
Their Antioxidant Activity
Mehdi Khoobia, Ali Ramazanib, Zahra Hojjatib, Raheleh Shakeric,
Mehdi Khoshneviszadehbd, Susan Kaboudanian Ardestanic, Abbas
Shafieeb, Alireza Foroumadia & Sang Woo Jooe
a Department of Medicinal Chemistry, Faculty of Pharmacy and
Pharmaceutical Sciences Research Center, Tehran University of
Medical Sciences, Tehran 14176, Iran
b Department of Chemistry, University of Zanjan, P.O. Box
45195-313, Zanjan, Iran
c Institute of Biochemistry and Biophysics (I.B.B.), University of
Tehran, P.O. Box 13145-1384, Tehran, I.R. Iran
d Medicinal & Natural Products Chemistry Research Center, Shiraz
University of Medical Sciences, Shiraz 71345, Iran
e School of Mechanical Engineering, Yeungnam University, Gyeongsan
712-749, Republic of Korea
Accepted author version posted online: 30 Apr 2014.Published
online: 21 Oct 2014.
To cite this article: Mehdi Khoobi, Ali Ramazani, Zahra Hojjati, Raheleh Shakeri, Mehdi
Khoshneviszadeh, Susan Kaboudanian Ardestani, Abbas Shafiee, Alireza Foroumadi & Sang Woo Joo
(2014): Synthesis of Novel 4H-Chromenes Containing a Pyrimidine-2-Thione Function in the Presence
of Fe3O4 Magnetic Nanoparticles and Study of Their Antioxidant Activity, Phosphorus, Sulfur, and
Silicon and the Related Elements, DOI: 10.1080/10426507.2014.884094
To link to this article: http://dx.doi.org/10.1080/10426507.2014.884094
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Phosphorus, Sulfur, and Silicon, 189:1–10, 2014
Copyright ꢀC Taylor & Francis Group, LLC
ISSN: 1042-6507 print / 1563-5325 online
DOI: 10.1080/10426507.2014.884094
SYNTHESIS OF NOVEL 4H-CHROMENES CONTAINING
A PYRIMIDINE-2-THIONE FUNCTION IN THE PRESENCE
OF Fe3O4 MAGNETIC NANOPARTICLES AND STUDY
OF THEIR ANTIOXIDANT ACTIVITY
Mehdi Khoobi,1 Ali Ramazani,2 Zahra Hojjati,2 Raheleh
Shakeri,3 Mehdi Khoshneviszadeh,2,4 Susan Kaboudanian
Ardestani,3 Abbas Shafiee,2 Alireza Foroumadi,1
and Sang Woo Joo5
1Department of Medicinal Chemistry, Faculty of Pharmacy and Pharmaceutical
Sciences Research Center, Tehran University of Medical Sciences, Tehran 14176,
Iran
2Department of Chemistry, University of Zanjan, P.O. Box 45195-313, Zanjan, Iran
3Institute of Biochemistry and Biophysics (I.B.B.), University of Tehran, P.O. Box
13145-1384, Tehran, I.R. Iran
4Medicinal & Natural Products Chemistry Research Center, Shiraz University
of Medical Sciences, Shiraz 71345, Iran
5School of Mechanical Engineering, Yeungnam University, Gyeongsan 712-749,
Republic of Korea
GRAPHICAL ABSTRACT
Abstract The aim of the present work was to search for identification of novel reactive oxy-
gen species (ROS) scavengers by testing new fused chromenopyrimidinethiones, which was
synthesized using Fe3O4 nanoparticles. The new compounds were also tested for their cy-
totoxic activity. The obtained results showed that the incorporated pyrimidinethione moiety
significantly increase antioxidant activity. In conclusion, the study of the pharmacological
properties of the new chromenopyrimidinethiones allowed establishing new structure–activity
relationships for splitting antioxidant and cytotoxic activity of these compounds.
Keywords Fe3O4 nanoparticles; antioxidant; synthesis; cytotoxic activity; chromenopyrim-
idinethiones
INTRODUCTION
Nanotechnology has been one of the most active research areas in recent years.1
The reactivity of catalytic nanoparticles is largely determined by the energy of surface
Received 1 September 2013; accepted 26 November 2013.
Address correspondence to Ali Ramazani, Department of Chemistry, University of Zanjan, P.O. Box 45195-
313, Zanjan, Iran. E-mail: aliramazani@gmail.com or to Sang Woo Joo, School of Mechanical Engineering,
Yeungnam University, Gyeongsan 712-749, Republic of Korea. E-mail: swjoo@yu.ac.kr
1

2
M. KHOOBI ET AL.
atoms, which can be easily gauged by the number of neighboring atoms and by the bonding
modes and accompanying energies of small molecules to be transformed on the surfaces of
nanoparticles.1,2 Magnetic nanoparticles are a class of nanostructured materials of current
interest, due to the largely advanced technology and medical applications, envisioned or re-
alized. Among the various magnetic nanoparticles under investigation, Fe3O4 nanoparticles
(Fe3O4 NPs) are arguably the most extensively studied.3 The main characteristic of these
nanoparticles is the simple and convenient separation from a reaction media by magnetic
separation.4,5 Recently, magnetite nanoparticles were used as an efficient catalyst in many
organic transformations.6–8
Various reactive oxygen species (ROS), such as the hydroxyl radical and superoxide
anion radical, have been known to induce lipid peroxidation as well as to damage mem-
branes, proteins, and DNA. Enzymatic and nonenzymatic antioxidative defense systems can
remove ROS under normal conditions. Oxidative stress, however, occurring when antiox-
idant systems are inadequate and/or active oxygen species are overproduced can damage
the tissues and DNA, thus resulting in a progression of a number of human diseases such
as atherosclerosis,9 diabetes, inflammation, Alzheimer’s disease, and senescence. Hence,
antioxidants may stop the free-radical formation, or interrupt an oxidizing chain reac-
tion.10 This had attracted a great deal of research interest in therapeutic antioxidant-based
drugs formulations. The development of synthetic compounds, capable of scavenging free
radicals, has been a great success.11
The chromene (or benzopyran) moiety often appears as an important structural com-
ponent in both biologically active and natural compounds. Chromene fragments occur
in alkaloids, flavonoids, tocopherols, and anthocyanins.12 Moreover, functionally sub-
stituted chromenes have played increasing roles in synthetic approaches to promising
compounds in the field of biomedicinal chemistry. The current interest in 4H-chromene
derivatives bearing a nitrile functionality arises from their potential application as an-
timicrobial,13 antiviral,14 antitumor15 agents and in the treatment of human inflamma-
tory TNF α-medicated diseases, such as rheumatoid and psoriatic arthritis and of cancer
therapy.16
Previous studies showed that 2-amino-4H-chromene derivatives exert cytotoxic activ-
ity through the apoptosis induction mechanism.17,18 Another study demonstrated the antiox-
idant activity of pyrimidine thione derivatives.19 Thus, in continuation of a research program
to find a novel anticancer drug,20,21 in the present study we incorporate the pyrimidinethione
moiety into 2-amino-4H-chromene scaffold in order to achieve new 4H-chromenes con-
taining a pyrimidine-2-thione function and evaluation of their cytotoxic and antioxidant
activities.
Development of clean technologies to replace hazardous materials in reaction
media with environmentally friendly substances is an increasing interest in “green
chemistry.” Water is an obviously benign and inexpensive solvent to reduce pollu-
tion, cost, and tedious work-ups in synthetic methodologies. Bearing in mind the
usefulness and efficiency of Fe3O4 NPs in organic reactions22 and in connection with
our previous works on evaluation of biological active compounds,23,24 we decided
to explore an application of Fe3O4 NPs for the preparation of 2-amino-4-aryl-4H-
chromene-3-carbonitrile derivatives 5 with using multicomponent reaction25 of 1, 2,
and 4 (Scheme 1). Also, a novel series of chromenopyrimidinethione derivatives 8
were synthesized in order to develop novel antioxidant agents with low cytotoxicity
activity.

SYNTHESIS OF NOVEL 4H-CHROMENES
3
Ar1
O
N
OH
O
CN
CN
CN
CN
4
Fe3O4 magnetic nanoparticles
water, reflux
+
Fe3O4 magnetic nanoparticles
water, reflux
Ar1
H
Ar1
CN
N
NH2
5
1
2
3
Ar1
Ar1
O
Ar1 NH
N
6
S
5
C
CN
3
Ar2
S
Ar2
HN
C
N
+
N
10
1
Ar2
N
N
O
NH2
N
8
N
O
N
S
H
H
5
7
6
8
180 ppm
Ar1 NH
S
Ar2
N
O
N
H
N
9
158 ppm
Scheme 1 Synthetic method for the synthesis and atom numbering of chromenopyrimidinethiones 8.
RESULTS AND DISCUSSION
Chemistry
As outlined in Scheme 1, compounds 8 could be easily prepared by a rather conve-
nience one-pot, three-component reaction of an aromatic aldehyde 1, malononitrile 2, and
3-(dimethylamino)phenol 4 to afford the corresponding 2-amino-4-aryl-4H-chromene-3-
carbonitrile 5 in high yields. The reaction was carried out in aqueous media under reflux
condition using catalytic amounts of Fe3O4 magnetic nanoparticles (Fe3O4 MNPs) as a
green, high-efficient and recoverable magnetic nanocatalyst for very short reaction time.
This “one-pot” reaction protocol proceeds through formation of arylidenemalononitrile
intermediate 3. The catalytic performances of Fe3O4 MNPs were screened in aqueous
media and solvent-free conditions. The results show that without the catalyst, 21% yields
could be obtained after 10 h and water is a suitable accelerator of the reaction. A viscous
mixture was obtained under solvent-free media, which cause to difficult dispersion and
magnetic separation of the catalyst. Also, we noticed that reducing in the amount of the
catalyst (5 mg) decreased the yield of the reaction and no significant improvement was
obtained when the catalyst increased (see Table S1, Supplemental Materials). In the next
step, the chromene 5 was treated with isothiocyanates 6 in various solvent and catalyst.
Unfortunately, a mixture of product was obtained in the presence of DBU or DABCO as
catalyst in several solvents, as well as separation and purification of the target product 8
was difficult (Table S1). Only in the presence of pyridine as solvent and catalyst under
reflux condition, the target product was obtained without any byproduct, easily separated
by simple filtration and purified from crystallization. All of products were appropriately
characterized by spectral data. For example, compound 8c was fully characterized by IR,
1H, and 13C NMR spectra and MS. The mass spectrum of 8c displayed a molecular ion
signal at m/z 486 and an ion signal at m/z 393 indicating the loss of the aniline group. In

4
M. KHOOBI ET AL.
the 1H NMR spectrum of compound 8c, in addition to the aromatic protons of chromene
ring and those assigned to the phenyl rings (δ = 8.40–6.46 ppm), a sharp singlet due to
hydrogen in the chromene moiety on 5 position (5.10 ppm) were observed. Also three sin-
glets at 3.71, 3.67, and 2.89 ppm due to the two methoxy and dimethylamino groups were
assigned, respectively. The most important absorption band of 8c in the IR spectrum was
detected at 3384 cm−1 and attributed to the N H group stretching frequency. Absorption
band at 1570 cm−1 was associated with the C N stretching frequency. The 1H decoupled
13C NMR spectrum of 8c showed 23 distinct signals. In this spectrum, the methine of the
chromene moiety on 5 position resonated at δ = 87 and the signal for the C S was observed
at δ = 178 ppm, respectively. As indicated in Scheme 1, the corresponding chromenoth-
iazine derivatives 9 could be achieved in the aqueous reaction media. These structures of
compounds 8 and 9 can be easily distinguished by their 13C NMR spectra. In compound 8,
C S appeared around 180 ppm, while in compound 9, C N on 2 position should resonate
around 158 ppm. Since in the product, no signal was detected around 180 ppm, compound
9 was consequently ruled out (see Scheme 1 and Table 1).
Biology
The antioxidant activities of the synthesized chromenopyrimidinethiones (8a-l) were
evaluated by two in vitro methods in order to compare the results and to establish some
structure antioxidant activity relationships for each method. The evaluation study was car-
ried out at various concentrations and in comparison with the standard antioxidants. The
DPPH radical scavenging activity assay is a simple method for measuring the antioxidant
ability to trap free radicals. The scavenging effects of chromenopyrimidinethiones (8a-l)
Table 1 Synthesis of 2-amino-4-aryl-4H-chromene-3-carbonitrile derivatives 5 and chromenopyrimidinethiones
8
Compounds
Ar1
Ar2
Yield (%)a
Time
mp
Ref.
26
26
26
26
26
26
26
5a
5b
5c
5d
5e
5f
5g
8a
8b
8c
8d
8e
8f
8g
8h
8i
8j
8k
8l
2,5-dimethoxyphenyl
3,4-dimethoxyphenyl
2,3-dimethoxyphenyl
3,4,5-trimethoxyphenyl
2-bromophenyl
—
—
—
—
—
78
73
80
71
68
73
77
58
66
48
59
62
68
63
54
71
61
64
72
30 min
35 min
45 min
30 min
50 min
45 min
50 min
24 h
24 h
24 h
24 h
24 h
24 h
24 h
24 h
24 h
24 h
24 h
24 h
126–128
159–161
107–109
182–184
199–201
177–179
207–208
291–293
272–274
>300
288–290
277–279
273–275
267–269
224–226
280–282
290–292
>300
3- bromophenyl
2,3-dichlorophenyl
—
—
2,5-dimethoxyphenyl
2,5-dimethoxyphenyl
3,4-dimethoxyphenyl
3,4-dimethoxyphenyl
3,4-dimethoxyphenyl
3,4-dimethoxyphenyl
2,3-dimethoxyphenyl
2,3-dimethoxyphenyl
3,4,5-trimethoxyphenyl
3,4,5-trimethoxyphenyl
3,4,5-trimethoxyphenyl
3,4,5-trimethoxyphenyl
2-bromophenyl
2-chlorophenyl
Ph
—
—
—
—
—
—
—
—
—
—
—
—
2-chlorophenyl
2-bromophenyl
2-fluorophenyl
2-chlorophenyl
2-bromophenyl
2-fluorophenyl
2-bromophenyl
2-chlorophenyl
Phenyl
>300
aIsolated yield.

SYNTHESIS OF NOVEL 4H-CHROMENES
5
are shown in Table S3 (Supplemental Materials). Two controls, ascorbic acid and trolox,
are included. As shown in Table S3, all pyrimidinethione derivatives 8a-l exhibited sig-
nificant DPPH radical scavenging activity against their precursors with IC50 values less
than 42 μg/mL. For example, a nine-fold enhancement in the radical scavenging activity
of compound 5d was observed by the addition of pyrimidinethione moiety to chromene
scaffold in 8l.
The ABTS assay is a widely used method for measuring the antioxidant ability to trap
free radicals. The ABTS radical cation scavenging capacity of chromenopyrimidinethiones
(8a-l) demonstrated all compounds have good antioxidant ability to trap free radicals. The
IC50 values of synthesized compounds were in the range of 21.28–72.98 μg/mL. Among the
chromenopyrimidinethiones, compound 8l exhibited the most potent antioxidant activity
in both DPPH and ABTS methods. Although compounds 8l was not as potent as reference
drug Trolox, but its antioxidant activity could be considered as attendant property of the
title compound.
The IC50 values of target compounds 8 against PC3 and HepG2 cells in Table S3
revealed that all compounds showed no growth inhibitory activity (IC50 > 100 μg/mL)
against two tested cell lines.
CONCLUSION
New chromene derivatives incorporating the pyrimidine thione moiety were synthe-
sized. The new compounds were evaluated for the antioxidant and cytotoxic activity. The
results showed that most of the chromenopyrimidinethiones had a high degree of potency in
scavenging activity against the DPPH and ABTS radicals and showed no cytotoxic activity
against cancer cell lines.
MATERIALS AND METHODS
Experimental
All commercially available reagents were used without further purification. Column
chromatography was carried out on silica gel (70–230 mesh). TLC was conducted on
silica gel 250 micron, F254 plates. Melting points were measured on a Kofler hot-stage
apparatus and are uncorrected. The IR spectra were taken using Nicolet FTIR Magna 550
spectrographs (KBr disks). The morphological analysis by X-ray diffraction was performed
on XPert MPD advanced diffractometer using Cu (Kα) radiation (wavelength: 1.5406 A◦)
at room temperature in the range of 2θ from 1 to 10◦ with a scanning rate of 0.02◦.S−1
.
The magnetic properties of samples were detected at room temperature using vibrating
sample magnet—ometer (VSM, Lake Shore 7410). The particle size and morphology of
the surfaces of Fe3O4 NPs27 were analyzed by a scanning electron microscopy (VEGAII
TESCAN) with an acceleration voltage of 15 kV. The disc was pasted with copper tape,
and the sample was dispersed over the tape. The disc was coated with gold in an ionization
chamber. 1H NMR spectra were recorded on a Bruker 400 or 500 MHz NMR instruments.
The chemical shifts (δ) and coupling constants (J) are expressed in parts per million
and hertz, respectively. Mass spectra of the products were obtained with a HP (Agilent
technologies) 5937 Mass Selective Detector. Elemental analyses were carried out by a
CHN-Rapid Heraeus elemental analyzer. The results of elemental analyses (C, H, N) were
within 0.4% of the calculated values.

6
M. KHOOBI ET AL.
Preparation of Fe3O4 Magnetic Nanoparticles. The coprecipitation method
was used to prepare the Fe3O4 NPs:28 FeCl3·6H2O (13.0 g) and FeCl2·4H2O (4.8 g) in
a 1:2 molar ratio were dissolved in distilled water (200 mL) under nitrogen atmosphere
with vigorous stirring. As the solution was heated to 70◦C, NH3·H2O (28 wt%, 25 mL)
was added dropwise to the solution under vigorous stirring and the reaction was allowed
to proceed for 5 h at 70◦C, and then the temperature was increased to 85◦C to vapor the
residual NH3. A more complete characterization is presented in the Supplemental Materials
(Figures S1–S4).
General Procedure for the Preparation of 2-Amino-4-aryl-4H-chromene-
3-carbonitrile Derivatives 5. A mixture of an aromatic aldehyde 1 (1.0 mmol), mal-
ononitrile 2 (1.2 mmol), Fe3O4 MNPs (10.0 mg), and water (5 mL) was heated under reflux
conditions for a few minutes to afford the corresponding intermediate 3. To this stirred
mixture, 3-(dimethylamino)phenol 4 (1.0 mmol) was added and the reaction mixture was
refluxed for the length of time as indicated in Table S1 (Supplemental Materials). The
progress of the reaction was monitored by TLC. After completion of the reaction, ethyl
acetate was added and the catalyst was easily separated from the reaction mixture with
an external magnet and washed twice with ethyl acetate. The combined organic layers
were concentrated in vacuum and the resulting residue was purified by recrystallization
from ethanol. Selected spectra for 8b and 8g are presented in the Supplemental Materials
(Figures S5–S9).
General Procedure for the Preparation of Compounds 8. A mixture of ap-
propriate2-amino-4-aryl-4H-chromene-3-carbonitrile5 (1.0mmol) andaryl isothiocyanate
(12.0 mmol) in dry pyridine (15 mL) was refluxed for 24 h. The progress of reaction was
followed by TLC. After completion of the reaction, the mixture was allowed to cool, washed
twice with n-hexane and finally the resulting residue was purified by recrystallization from
ethanol.
3-(2-Bromophenyl)-5-(2,5-dimethoxyphenyl)-8-(dimethylamino)-4-
imino-3,4-dihydro-1H-chromeno[2,3-d]pyrimidine-2(5H)-thione
(8a). White
solid, Yield (52%), mp = 291–293◦C; IR (KBr) (νmax, cm−1): 3372 (NH) and 1627 (C N).
1H NMR (500 MHz, CDCl3): δ 2.88 (s, 6H, N(Me)2), 3.62 (s, 6H, 2OMe), 5.21 (s, H4
chromene), 6.44–6.87 (m, 6H, 3H chromene, and 3H phenyl), 7.38–7.68 (m, 4H, N-Ph).
Anal. Calcd. for C27H25BrN4O3S (565): C, 57.35; H, 4.46; N, 9.91. Found: C, 57.42; H,
4.10; N, 9.52%.
3-(2-Chlorophenyl)-5-(2,5-dimethoxyphenyl)-8-(dimethylamino)-4-
imino-3,4-dihydro-1H-chromeno[2,3-d]pyrimidine-2(5H)-thione
(8b). White
solid, Yield (59%), mp = 272–274◦C; IR (KBr) (νmax, cm−1): 3371 (NH) and 1630 (C N).
1H NMR (500 MHz, CDCl3): δ 2.86 (s, 6H, N(Me)2), 3.63 (s, 6H, 2OMe), 5.22 (s, H4
chromene), 6.45–6.87 (m, 6H, 3H chromene, and 3H phenyl), 7.27–7.64 (m, 4H, N-Ph).
13C NMR (125 MHz, CDCl3): δ 55.2, 56.3, 56.5, 87.0, 99.2, 109.2, 109.9, 112.0, 113.3,
113.4, 115.4, 115.8, 129.0, 129.1, 130.7, 131.1, 133.1, 133.3, 135.7, 150.1, 150.3, 150.6,
153.2, 153.4, 160.6, 177.8. Anal. Calcd. for C27H25ClN4O3S (521): C, 62.24; H, 4.84; N,
10.75. Found: C, 62.41; H, 5.02; N, 10.68%.
5-(3,4-Dimethoxyphenyl)-8-(dimethylamino)-4-imino-3-phenyl-3,4-
dihydro-1H-chromeno[2,3-d]pyrimidine-2(5H)-thione (8c). White solid, Yield
1
(40%), mp > 300◦C; IR (KBr) (νmax, cm−1): 3384 (NH) and 1616 (C N). H NMR
(500 MHz, CDCl3): δ 2.86 (s, 6H, N(Me)2), 3.63 (s, 6H, 2OMe), 5.22 (s, H4 chromene),
6.46 (s, 1H, H8 chromene), 6.51 (d, 1H, J = 7.6 Hz H6 chromene), 6.74 (d, 1H, J = 7.6 Hz,

SYNTHESIS OF NOVEL 4H-CHROMENES
7
H5 phenyl), 6.84 (d, 1H, J = 7.6 Hz, H6 phenyl), 6.99 (s, 1H, H2 phenyl), 7.00 (d, 1H, J =
7.6 Hz, H5 chromene), 7.03–8.40 (m, 5H, N-Ph). MS (EI, 70 eV): m/z (%) = 486 (100),
426 (35), 393 (41), 363 (52), 349 (58), 335 (23), 290 (76), 256 (73), 214 (64), 135 (20), 93
(29), 77 (23). Anal. Calcd. for C27H26N4O3S (486): C, 66.65; H, 5.39; N, 11.51. Found:
C, 66.42; H, 5.12; N, 11.68%.
3-(2-Chlorophenyl)-5-(3,4-dimethoxyphenyl)-8-(dimethylamino)-4-
imino-3,4-dihydro-1H-chromeno[2,3-d]pyrimidine-2(5H)-thione
(8d). White
solid, Yield (52%), mp = 288–290◦C; IR (KBr) (νmax, cm−1): 3372 (NH) and 1619 (C N).
1H NMR (500 MHz, CDCl3): δ 3.13 (s, 6H, N(Me)2), 3.67 (s, 6H, 2OMe), 5.13 (s, H4
chromene), 6.45–6.98 (m, 6H, 3H chromene, and 3H phenyl), 7.29–7.64 (m, 4H, N-Ph).
Anal. Calcd. for C27H25ClN4O3S (521): C, 62.24; H, 4.84; N, 10.75. Found: C, 62.12; H,
4.73; N, 10.68%.
3-(2-Bromophenyl)-5-(3,4-dimethoxyphenyl)-8-(dimethylamino)-4-
imino-3,4-dihydro-1H-chromeno[2,3-d]pyrimidine-2(5H)-thione
(8e). White
solid, Yield (57%), mp = 277–279◦C; IR (KBr) (νmax, cm−1): 3373 (NH) and 1619 (C N).
1H NMR (500 MHz, CDCl3): δ 2.88 (s, 6H, N(Me)2), 3.66 (s, 6H, 2OMe), 5.14 (s, H4
chromene), 6.46–6.99 (m, 6H, 3H chromene, and 3H phenyl), 7.39–7.78 (m, 4H, N-Ph).
Anal. Calcd. for C27H25BrN4O3S (565): C, 57.35; H, 4.46; N, 9.91. Found: C, 57.42; H,
4.42; N, 9.68%.
3-(2-Fluorophenyl)-5-(3,4-dimethoxyphenyl)-8-(dimethylamino)-4-
imino-3,4-dihydro-1H-chromeno[2,3-d]pyrimidine-2(5H)-thione
(8f). White
solid, Yield (65%), mp = 273–275◦C; IR (KBr) (νmax, cm−1): 3398 (NH) and 1617
1
(C N). H NMR (500 MHz, CDCl3): δ 2.89 (s, 6H, N(Me)2), 3.67 (s, 3H, OMe), 3.71
(s, 3H, OMe), 5.14 (s, H4 chromene), 6.45–7.12 (m, 6H, 3H chromene, and 3H phenyl),
7.24–7.53 (m, 4H, N-Ph). Anal. Calcd. for C27H25FN4O3S (504): C, 64.27; H, 4.99; N,
11.10. Found: C, 64.42; H, 5.02; N, 11.18%.
3-(2-Chlorophenyl)-5-(2,3-dimethoxyphenyl)-8-(dimethylamino)-4-
imino-3,4-dihydro-1H-chromeno[2,3-d]pyrimidine-2(5H)-thione
(8g). White
solid, Yield (55%), mp = 267–269◦C; IR (KBr) (νmax, cm−1): 3372 (NH) and 1627
1
(C N). H NMR (500 MHz, CDCl3): δ 2.89 (s, 6H, N(Me)2), 3.64 (s, 3H, OMe), 3.74
(s, 3H, OMe), 5.25 (s, H4 chromene), 6.46–6.97 (m, 6H, 3H chromene, and 3H phenyl),
7.24–7.63 (m, 4H, N-Ph). Anal. Calcd. for C27H25ClN4O3S (521): C, 62.24; H, 4.84; N,
10.75. Found: C, 62.42; H, 5.02; N, 10GPSS-2013-0279.68%.
3-(2-Bromophenyl)-5-(2,3-dimethoxyphenyl)-8-(dimethylamino)-4-
imino-3,4-dihydro-1H-chromeno[2,3-d]pyrimidine-2(5H)-thione
(8h). White
solid, Yield (49%), mp = 224–226◦C; IR (KBr) (νmax, cm−1): 3395 (NH) and 1615 (C N).
1H NMR (500 MHz, CDCl3): δ 2.89 (s, 6H, N(Me)2), 3.63 (s, 3H, OMe), 3.74 (s, 3H,
OMe), 5.25 (s, H4 chromene), 6.47–6.98 (m, 6H, 3H chromene and 3H phenyl), 7.24–7.78
(m, 4H, N-Ph). Anal. Calcd. for C27H25BrN4O3S (565): C, 57.35; H, 4.46; N, 9.91. Found:
C, 57.42; H, 4.42; N, 9.68%.
3-(2-Fluorophenyl)-5-(3,4,5-trimethoxyphenyl)-8-(dimethylamino)-4-
imino-3,4-dihydro-1H-chromeno[2,3-d]pyrimidine-2(5H)-thione
(8i). White
solid, Yield (68%), mp = 280–282◦C; IR (KBr) (νmax, cm−1): 3398 (NH) and 1617
1
(C N). H NMR (500 MHz, CDCl3): δ 2.88 (s, 6H, N(Me)2), 3.57 (s, 3H, OMe), 3.70
(s, 6H, 2OMe), 5.12 (s, H4 chromene), 6.50–7.12 (m, 5H, 3H chromene and 2H phenyl),
7.30–7.52 (m, 4H, N-Ph). Anal. Calcd. for C28H27FN4O4S (534): C, 62.91; H, 5.09; N,
10.48. Found: C, 62.42; H, 5.12; N, 10.68%.

8
M. KHOOBI ET AL.
3-(2-Bromophenyl)-5-(3,4,5-trimethoxyphenyl)-8-(dimethylamino)-4-
imino-3,4-dihydro-1H-chromeno[2,3-d]pyrimidine-2(5H)-thione
(8j). White
solid, Yield (59%), mp = 290–292◦C; IR (KBr) (νmax, cm−1): 3390 (NH) and 1616
1
(C N). H NMR (500 MHz, CDCl3): δ 2.88 (s, 6H, N(Me)2), 3.57 (s, 3H, OMe), 3.69
(s, 6H, 2OMe), 5.13 (s, H4 chromene), 6.51–7.03 (m, 5H, 3H chromene, and 2H phenyl),
7.39–7.74 (m, 4H, N-Ph). Anal. Calcd. for C28H27BrN4O4S (595): C, 56.47; H, 4.57; N,
9.41. Found: C, 56.42; H, 4.52; N, 9.61%.
3-(2-Chlorophenyl)-5-(3,4,5-trimethoxyphenyl)-8-(dimethylamino)-4-
imino-3,4-dihydro-1H-chromeno[2,3-d]pyrimidine-2(5H)-thione
(8k). White
solid, Yield (61%), mp > 300◦C; IR (KBr) (νmax, cm−1): 3389 (NH) and 1616 (C N). 1H
NMR (500 MHz, CDCl3): δ 2.88 (s, 6H, N(Me)2), 3.57 (s, 3H, OMe), 3.70 (s, 6H, 2OMe),
5.13 (s, H4 chromene), 6.46–7.13 (m, 5H, 3H chromene, and 2H phenyl), 7.34–7.64 (m,
4H, N-Ph). Anal. Calcd. for C28H27ClN4O4S (551): C, 61.03; H, 4.94; N, 10.17. Found:
C, 60.92; H, 5.02; N, 10.28%.
5-(3,4,5-Trimethoxyphenyl)-8-(dimethylamino)-4-imino-3-phenyl-3,4-
dihydro-1H-chromeno[2,3-d]pyrimidine-2(5H)-thione (8l). White solid, Yield
1
(42%), mp > 300◦C; IR (KBr) (νmax, cm−1): 3390 (NH) and 1615 (C N). H NMR
(500 MHz, CDCl3): δ 2.88 (s, 6H, N(Me)2), 3.58 (s, 3H, OMe), 3.71 (s, 6H, 2OMe), 5.12
(s, H4 chromene), 6.46–7.03 (m, 5H, 3H chromene, and 2H phenyl), 7.12–7.54 (m, 5H,
N-Ph). Anal. Calcd. for C28H28N4O4S (516): C, 65.10; H, 5.46; N, 10.85. Found: C, 65.42;
H, 5.32; N, 10.68%.
Biological Evaluation
Antioxidant Activity. ABTS·+ radical scavenging assay
ABTS radical scavenging activity was measured using the method of Pennycooke
et al. with some modifications.29 ABTS stock solution was prepared by reacting 7 mM
2,2ꢁ-azinobis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) with 2.45 mM ammonium
persulfate in dark at room temperature for 12–16 h.
DPPH radical scavenging assay
The DPPH free-radical scavenging activity of the synthetic compounds was assayed
on the basis of Brand-William et al. with some modifications.30
Cytotoxicity Study. The in vitro anti-cancer activity of target compounds 8a-l was
determined against PC3 (prostate cancer cell lines) and HepG2 (human liver carcinoma)
cells using MTT (3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method.31
Additional details are provided in the Supplemental Materials.
FUNDING
This work is funded by the grant 2011-0014246 of the National Research Foundation
of Korea. The authors thank Tehran Medical Sciences, Zanjan, Tehran and Shiraz Medical
Sciences Universities for the support and guidance.
SUPPLEMENTAL MATERIAL
Supplementary data for this article can be accessed on the publisher’s website,
www.tandfonline.com/gpss

SYNTHESIS OF NOVEL 4H-CHROMENES
9
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