Synthesis and biological activity assays of some new N1-(flavon-7-yl) amidrazone derivatives and related congeners

Article abstract of DOI:10.1016/j.ejmech.2012.04.028

A series of new N1-(flavon-7-yl)amidrazones incorporating N-piperazines and related congeners were synthesized by reacting the hydrazonoyl chloride derived from 7-aminoflavone and 7-amino-2-methylchromen-4-one with the appropriate piperazine. The chemical structures of the newly prepared compounds were confirmed by elemental analyses, ¹H NMR, ¹³C NMR, and ESI-HRMS spectral data. The antitumor activity of these compounds was evaluated on breast cancer (MCF-7 and T47D) and Leukemic (K562) cell lines by a cell viability assay utilizing the tetrazolium dye 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT). Although with varying degrees, a significant growth inhibitory and cytotoxic effect was observed on all three cancer cell lines. Among the compounds tested compounds, 5a, 15a, and 18b, were the most active against T47D cell line with IC₅₀ values of 1.42, 1.92, and 2.92 μM, respectively. By using other cancer cell lines and with further characterization of their biological mechanism of action, these compounds could prove to be useful candidates as anticancer drugs.

Full text of DOI:10.1016/j.ejmech.2012.04.028

European Journal of Medicinal Chemistry 54 (2012) 65e74
Contents lists available at SciVerse ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Synthesis and biological activity assays of some new N1-(flavon-7-yl)amidrazone
derivatives and related congeners
Marwa N. Abu-Aisheh a, Mohammad S. Mustafa a, Mustafa M. El-Abadelah a, Randa G. Naffa c,
,
Said I. Ismail c, Malek A. Zihlif b, Mutasem O. Taha d, Mohammad S. Mubarak a
*
a Department of Chemistry, Faculty of Science, The University of Jordan, Amman 11942, Jordan
b Department of Pharmacology, Faculty of Medicine, The University of Jordan, Amman 11942, Jordan
c Department of Biochemistry, Faculty of Medicine, The University of Jordan, Amman 11942, Jordan
d Drug Discovery Unit, Faculty of Pharmacy, The University of Jordan, Amman 11942, Jordan
h i g h l i g h t s
g r a p h i c a l a b s t r a c t
< A series of new N1-(flavon-7-yl)
amidrazones
incorporating
N-
piperazines and related congeners
were synthesized.
< Some of the prepared compounds
exhibited high potent activity
against breast cancer (MCF-7 and
T47D) and Leukemic (K562) cell
lines.
< The best results were obtained by
compounds 5a, 15a, and 18b against
T47D cell line with IC50 values of
1.42, 1.92, and 2.92 mM, respectively.
a r t i c l e i n f o
a b s t r a c t
Article history:
A series of new N1-(flavon-7-yl)amidrazones incorporating N-piperazines and related congeners were
synthesized by reacting the hydrazonoyl chloride derived from 7-aminoflavone and 7-amino-2-
methylchromen-4-one with the appropriate piperazine. The chemical structures of the newly
prepared compounds were confirmed by elemental analyses, 1H NMR, 13C NMR, and ESI-HRMS spectral
data. The antitumor activity of these compounds was evaluated on breast cancer (MCF-7 and T47D) and
Leukemic (K562) cell lines by a cell viability assay utilizing the tetrazolium dye 3-(4,5-dimethylthiazol-2-
Received 21 February 2012
Received in revised form
18 April 2012
Accepted 19 April 2012
Available online 15 May 2012
yl)-2,5-diphenyltetrazolium bromide (MTT). Although with varying degrees,
inhibitory and cytotoxic effect was observed on all three cancer cell lines. Among the compounds tested
compounds, 5a, 15a, and 18b, were the most active against T47D cell line with IC50 values of 1.42, 1.92,
and 2.92 mM, respectively. By using other cancer cell lines and with further characterization of their
a significant growth
Keywords:
7-aminoflavone
7-amino-2-methylchromen-4-one
4-oxo-N-(chromen-7-yl)hydrazonoyl
chloride
biological mechanism of action, these compounds could prove to be useful candidates as anticancer
1-piperazinyl amidrazones
Nitrile imine
Antitumor activity
drugs.
Ó 2012 Elsevier Masson SAS. All rights reserved.
1. Introduction
Chromones occur widely in nature, and have attracted much
synthetic interest because of their reactivity and biological activity
of their naturally-occurring representatives [1]. Chromones are
* Corresponding author.
well-known for their antioxidant activity [2], a property that stems
E-mail address: mmubarak@ju.edu.jo (M.S. Mubarak).
0223-5234/$ e see front matter Ó 2012 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejmech.2012.04.028

66
M.N. Abu-Aisheh et al. / European Journal of Medicinal Chemistry 54 (2012) 65e74
Scheme 1. Synthesis of compounds 4e19.
from their ability to neutralize active forms of oxygen and to cut off
free radical processes [3]. They also possess low mammalian toxicity
and are present in large amounts in the diet of humans due to their
origin in plants [4], especially in fruits and vegetables [5]. Flavo-
noids, a group of benzopyrone derivatives [6], have been recognized
as one of the largest and most widespread class of plant constituents
occurring throughout the plant kingdom [7]. A great number of
biological aspects were reported for flavones [8], such as biocidal [9],
immune-stimulatory [10], anti-ulcer [11], anti-estrogenic [12],
antitumor [13], anti-allergic [14], anti-inflammatory [15], anti-viral
[16], and anti-HIV [17] activities.
Examples include Khellin (1) [18] and 2,4-thiazolidenedione (2)
[19] that are chromone derivatives used as anti-spasmodic and
antidiabetic agents, respectively. These promising properties led to
numerous chemical works focusing on the synthesis and the
structural modifications of flavones [20].
Amino flavones are highly active molecules, wherein positions 5
and 7 are the most important and the most beneficial [21]. Flavo-
noids, bearing amino groups on the benzo- or pyranone ring have
been reported to be potential antineoplastic agents [22]. It is now
well established that such potency is mainly due to the ability of
these aminoflavones to be competitive inhibitors of certain
proteinetyrosine kinases with respect to ATP [23].
On the other hand, piperazine derivatives have drawn consid-
erable attention from organic and medicinal chemists. Piperazine-
based compounds have been employed as antibacterial [24], anti-
depressant [25], and antitumor [26] drugs, and as
a-adrenoceptor
antagonists [27]. Quite recently, several synthetic N-arylpiperazinyl
amidrazones were reported to display significant antitumor activity
against a number of human tumor cell lines [28]. Accordingly, we
envisaged to prepare a set of amidrazones incorporating flavone
moiety for evaluation of their antitumor activity.

M.N. Abu-Aisheh et al. / European Journal of Medicinal Chemistry 54 (2012) 65e74
67
2. Results and discussion
(MTT). Cultures of the breast cancer cell lines MCF-7 and the
Leukemic cell line K562 leukemia were treated with the target
2.1. Chemistry
compounds first at one concentration of 50 mM and the results are
shown in Table 1. In the MCF-7 screening test, ten compounds
showed anti-MCF-7 activity. Those compounds were able to reduce
the viability after 72 h to less than 50%. In the case of K562 cells,
only six compounds illustrated anti-K562 activity. Further we
determined the IC50 values for the potential compounds against the
MCF-7 and K562 (Table 2). We also explored the activity against one
additional breast cancer cell line called T47D. Interestingly, five
compounds showed high potency against MCF-7, whereby they
The hydrazonoyl chlorides 4a and 4b, required in this study,
were prepared via direct coupling of the respective flavon-7-
diazonium chloride or 2-methylchromenone-7-diazonium chlo-
ride with 3-chloropentane-2,4-dione in aqueous pyridine
(JappeKlingemann reaction) [29, 30] (Scheme 1). The latter 7-
diazonium chlorides are freshly prepared by diazotization of the
respective 7-amino-compounds (3a and 3b), (suspended in 6 N aq.
HCl).
Piperazine, N-substituted piperazines and related cyclic
secondary amine congeners, acting as nitrogen nucleophiles, are
expected to add readily onto N-(falvon-7-yl) or N-(2-
methylchromenone-7-yl)nitrile imines (the reactive 1,3-dipolar
species generated in situ from their corresponding hydrazonoyl
chloride precursors 4a and 4b in the presence of triethylamine) to
give the respective amidrazone adducts 5ae9a, 11ae19a and
7be18b (Scheme 1). This mode of nucleophilic addition reaction of
various nucleophiles onto 1,3-dipoles is well documented [31e39]
and several adducts related to 5e19 were obtained from the reac-
tion of amines with hydrazonoyl chlorides.
The newly synthesized compounds 4e19 were characterized by
elemental analyses, MS and NMR spectral data. These data, detailed
in the experimental part, are consistent with the suggested struc-
tures. Thus, the mass spectra display the correct molecular ion
peaks for which the measured high resolution (HRMS) data are in
good agreement with the calculated values. DEPT and 2D (COSY,
HMQC, HMBC) experiments showed correlations that helped in the
1H- and 13C-signal assignments to the different carbons and their
attached, and/or neighboring hydrogens.
scored IC50 values less than 10
more encouraging results were obtained. Here the number of
compounds that scored an IC50 of less 10 M increased to six and
the IC50 values were lower than those scored in the MCF-7 case
reaching in 5a and 15a cases an IC50 value of less than 2
mM (Table 2). On the T47D scenario,
m
mM
(Table 2). Finally, the IC50 values against the K562 cell line were also
encouraging, although only two compounds showed IC50 values
less than 10 mM. Importantly, compound 11a scored the lowest IC50
values against K562 although its potency against MCF-7 and T47D
was not the best (Table 2).
2.3. Structureeactivity relationship analysis and docking-based
explanation
A comparison of the compounds’ antitumor activity against
different cell lines is effectively presented in Tables 1and 2. The two
sets of compounds, 5ae9a and 11ae19a, and 7be18b differ in the
substitution on C-2; a phenyl group is present on C-2 in the first
group (5ae9a and 11ae19a), while a methyl group is connected to
C-2 in the second group (7be18b). The presence of the phenyl
group at C-2 apparently does make a difference for the anti-K562
activity since all compounds in the first group (5ae9a and
11ae19a) displayed moderate to excellent activity whereas none of
the compounds having a methyl substituent at C-2 (7be18b)
showed any activity against the K562 cells lines. The same series
of compounds were screened against breast cancer; results
revealed that more compounds of the flavone series (where there is
a phenyl group linked to C-2) and only few of the 2-methyl series
displayed activity as shown in Table 1. In addition, the IC50 values
for 5a,11a,12a and 13a, for the anti-breast cancer activity reveal the
importance of the piperazine moiety and to a lesser extent the
piperidine moiety. On the other hand, the situation is different for
the anti-leukemic activity, where the anti-K562 activity decreases
in the order 5a < 11a < 12a < 13a; this demonstrates the impor-
tance of the piperidine and piperazine moieties and to a lesser
extent for thiomorpholine ring system.
2.2. Biology
The antitumor activity of the synthesized compounds was
characterized by conducting cell viability assay using tetrazolium
dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
Table 1
Percentage cell survival of MCF-7 and K562 following 72 h exposure to 50
compounds. Doxorubicin is used as a positive control.
mM of all
Compound
MCF-7% survival ꢀ SD
K562% survival ꢀ SD
Doxorubicin
(5a)
(6a)
(7a)
(7b)
(8a)
(8b)
(9a)
(9b)
(10b)
(11a)
(11b)
(12a)
(12b)
(13a)
(13b)
(14a)
(14b)
(15a)
(15b)
(16a)
(16b)
(17a)
(17b)
(18a)
(18b)
23.6 ꢀ 1.67
10.28 ꢀ 0.02
7.98 ꢀ 0.24
11.90 ꢀ 0.90
7.69 ꢀ 0.07
5.18 ꢀ 0.01
9.77 ꢀ 0.46
15.88 ꢀ 0.02
77.42 ꢀ 0.04
83.27 ꢀ 6.10
92.92 ꢀ 5.75
68.27 ꢀ 0.05
84.38 ꢀ 4.02
78.25 ꢀ 0.08
44.58 ꢀ 2.71
105.35 ꢀ 6.71
19.83 ꢀ 0.02
114.34 ꢀ 3.09
94.56 ꢀ 3.75
85.53 ꢀ 5.21
64.85 ꢀ 0.08
80.03 ꢀ 0.05
60.86 ꢀ 0.09
90.78 ꢀ 2.11
72.62 ꢀ 0.11
90.41 ꢀ 5.12
83.90 ꢀ 3.85
74.47 ꢀ 6.24
42.19 ꢀ 0.07
79.40 ꢀ 3.43
69.53 ꢀ 0.02
44.32 ꢀ 0.02
75.90 ꢀ 4.45
14.36 ꢀ 0.14
87.05 ꢀ 5.74
82.64 ꢀ 0.06
18.01 ꢀ 0.01
65.13 ꢀ 3.87
66.77 ꢀ 6.19
11.80 ꢀ 1.56
76.73 ꢀ 7.93
75.64 ꢀ 0.04
75.58 ꢀ 0.01
65.84 ꢀ 0.03
88.54 ꢀ 0.05
43.94 ꢀ 0.08
66.42 ꢀ 0.04
104.56 ꢀ 2.51
20.27 ꢀ 0.01
42.80 ꢀ 0.05
83.70 ꢀ 0.10
20.56 ꢀ 0.01
Table 2
Effects of compounds that have shown potential activity on the screening assay
T47D, MCF-7 and K562. Doxorubicin is used as a positive control.
Compound IC50 T47D (
mM) ꢀ SD IC50 MCF-7 (
m
M) ꢀ SD IC50 K562 ( M) ꢀ SD
m
Doxorubicin 0.33 ꢀ 0.05
0.31 ꢀ 0.01
5.91 ꢀ 1.61
22.37 ꢀ 4.56
56.79 ꢀ 8.88
13.56 ꢀ 1.82
21.59 ꢀ 5.87
e
1.41 ꢀ 0.31
(5a)
1.42 ꢀ 0.13
15.76 ꢀ 1.38
53.37 ꢀ 4.03
11.05 ꢀ 0.61
8.79 ꢀ 0.80
e
5.02 ꢀ 0.78
(6a)
(7a)
(9a)
16.15 ꢀ 4.17
35 ꢀ 1.06
e
(11a)
(12a)
(15a)
(15b)
(17a)
(17b)
(18a)
(18b)
2.56 ꢀ 0.57
20.18 ꢀ 1.41
1.92 ꢀ 0.35
3.46 ꢀ 1.00
4.31 ꢀ 0.54
19.76 ꢀ 2.06
e
2.75 ꢀ 0.73
6.33 ꢀ 0.21
8.75 ꢀ 1.38
57.75 ꢀ 5.57
e
e
e
e
e
14.07 ꢀ 0.52
2.92 ꢀ 0.94
3.39 ꢀ 0.88
e

68
M.N. Abu-Aisheh et al. / European Journal of Medicinal Chemistry 54 (2012) 65e74
increased in many cases, a finding that emphasizes the importance
of the piperazine ring. Interestingly, those compounds that have
scored lower IC50 values than compound 5a have longer side
chains, for example, compound 8a has an extra carbon and
compound 18b has an extra methoxy group.
The potent inhibitory effects of our new compounds against
K562 and MCF-7 cancer cell lines, which over-express bcr/abl and
EGFR tyrosine kinases, respectively [40,41], combined with the
apparent pharmacophoric commonalities between these
compounds and the anticancer agent imatinib (in particular the
benzo-flavones 5ae9a and 11ae19a), prompted us to anticipate
that their observed anticancer properties are attributable to their
abilities to effectively bind and block oncogenic tyrosine kinases,
particularly bcr/abl. Fig. 1 compares how imatinib binds within the
ATP binding pocket of bcr/abl (PDB code: 1IEP, resolution 2.1 Å)
with the way 6a (the most active analogue, as in Table 1) docks into
the binding pocket of the same protein. Clearly from the figure,
positioning the pyridinylepyrimidine fragment of imatinib within
the aromaticehydrophobic pocket of the side chains of Phe382,
Tyr253 and Phe317 (Fig. 1a), compares to fitting the phenyl
substituent of 6a into the same pocket (Fig. 1b), i.e., via p-stacking
interactions. Similarly, hydrogen-bonding interactions connecting
the amidic linker of imatinib with the carboxylic acid side chain of
Glu286 and the peptidic NH of Asp381 correlate well with
hydrogen-bonding interactions connecting the amidrazone frag-
ment of 6a with the same amino-acid residues. Similar analogy can
be noticed between hydrogen-bonding interactions connecting the
hydroxyl of Thr315 with the aromatic NH of imatinib (Fig. 1a) and
the flavone oxygen of 6a (Fig. 1b). Furthermore, hydrophobic
stacking of the methylbenzene linker of imatinib within a narrow
corridor comprised of the CH3Sꢁ of Met290 and the ꢁ(CH2)4ꢁ of
Lys271 (Fig. 1a) compares to fitting the flavone ring of 6a within the
same corridor (Fig. 1b). Finally, the apparent electrostatic attraction
connecting the piperazine ring of imatinib with the carboxylate
side chain of Asp381 (Fig.1a) corresponds to electrostatic attraction
connecting the terminal piperazine nitrogen of 6a with the same
carboxylate group in the binding pocket.
3. Conclusion
In summary, we have synthesized a novel series of N1-(flavon-
7-yl)amidrazones incorporating N-piperazines and related conge-
ners through the reaction of the hydrazonoyl chloride derived from
7-aminoflavone and 7-amino-2-methylchromen-4-one with the
appropriate piperazine. The prepared compounds were tested
in vitro for their antitumor activity against breast cancer (MCF-7
and T47D) and Leukemic (K562) cell lines. The results revealed that
a number of the synthesized compounds exerted significant anti-
proliferative activity with the aforementioned cancer cell lines,
particularly compounds 5a, 15a, and 18b, with IC50 values of 1.42,
1.92, and 2.92
mM, respectively, against T47D cell line. The same
Fig. 1. (A) X-ray crystallographic structure of imatinib co-crystallized within c-abl
kinase domain (PDB code: 1IEP, resolution 2.1 Å), (B) compound 6a docked within the
same binding pocket, (C) superposition of the co-crystalized structure of imatinib over
the docked structure of 6a.
compounds have IC50 values of 5.91, 2.75, and 3.39
m
M, respectively
against MCF-7. In addition, moderate antitumor activity was dis-
played by all of the prepared compounds against those cell lines.
These findings would encourage us to do further studies and testing
that prove the usefulness of the prepared compounds as candidate
anticancer agents.
Similarly, for the anti-leukemic activity, compounds 5a and 11a,
are the most active; this finding drove us to conclude that leaving
the piperazine ring unsubstituted is better for the anti-K562
activity while for breast cancer, the anti-T47D activity resembles
that of the K562 activity. However, the results are more multifac-
eted in MCF-7, where some of the side chains resulted in negative
consequences on the anti-breast cancer activity, and others have
improved the activity. For example, the IC50 for compound 5a has
4. Experimental
4.1. Materials and equipments
All chemicals used were obtained from commercial sources and
were used as received without further purification. 7-aminoflavone,
7-amino-2-methylchromenone, piperidine, morpholine, thiomor-

M.N. Abu-Aisheh et al. / European Journal of Medicinal Chemistry 54 (2012) 65e74
69
pholine, piperazine, N-alkylpiperazines, N-arylpiperazines, and N-
acylpiperazine were purchased from Acros. Silica gel for column
chromatography was received from MachereyeNagel GmbH & Co
(Germany). Melting points (uncorrected) were determined on
a Stuart scientific melting point apparatus in open capillary tubes.1H
and 13C NMR spectra were recorded on a 300 MHz spectrometer
(Bruker DPX-300) with TMS as the internal standard. Chemical shifts
J ¼ 8.7,1.8 Hz,1H, H-6), 8.19 (d, J ¼ 8.7 Hz,1H, H-5), 9.56 (s,1H, NeH).
13C NMR (75 MHz, DMSO-d6):
20.7 (CH3-2), 26.1 (O]CeCH3),100.9
d
(C-8), 110.1 (C-3), 112.5 (C-6), 118.1 (C-4a), 127.5 (C-5), 145.4 (C-7),
147.8 (eC]N),158.0 (C-8a),165.5 (C-2),177.8 (C-4),189.1 (O]CeMe).
HRMS (ESI) m/z: Calcd for C13H12ClN2O3 [M þ H]þ 279.05364; found
279.05310. Anal. Calcd for C13H11ClN2O3 (278.69 g/mol): C, 56.03, H,
3.98, N, 10.05, found: C, 56.14, H, 4.02, N, 9.88.
are expressed in d
units; 1He1H, 1HeF and 13CeF coupling constants
are given in Hertz. High resolution mass spectra (HRMS) were
acquired by electrospray ionization (ESI) technique with the aid of
Bruker APEX-2 instrument. The samples were dissolved in acetoni-
trile, diluted in spraysolution (methanol/water 1:1 v/v þ 0.1% formic
4.3. General procedure of amidrazones preparation (5e19)
To a cold suspension (ꢁ10e0 ꢂC) of 1.80 mmol of 4 in 20.0 mL of
ethanol was added, with stirring,
a solution of appropriate
acid) and infused using a syringe pump with a flow rate of 2
mL/min.
secondary amine (2.0 mmol) and triethylamine (3 mL) in 5 mL of
ethanol. Stirring was continued at 0e5 ꢂC for 2e4 h, and then at
ambient temperature for additional 2 h then the solution poured
onto water (100 mL), the resulting crude solid product was
collected by suction filtration, washed with water, dried and puri-
fied on preparative silica gel TLC plates. Using the same general
procedure, the following compounds were prepared:
External calibration was conducted using arginine cluster in a mass
range m/z 175e871. Elemental analyses were performed on a Euro-
Vector Elemental Analyzer (EA3000A).
4.2. General procedure for preparation of N-([2-substituted]-4-oxo-
4H-chromen-7-yl)-2-oxo-propanehydrazonoyl chlorides (4a,b)
The title compounds were prepared by the following procedures:
4.3.1. 7-[2-(2-oxo-1-(piperazin-1-yl)propylidene)hydrazinyl]-2-ph
enyl-4H-chromen-4-one (5a)
Step (i). 7-aminoflavone 3a or 7-amino-2-methylchromen-4-
one 3b (0.10 mol) was dissolved in 6 N aqueous
hydrochloric acid (160 mL). To this solution was
added, dropwise, a solution of sodium nitrite (7.6 g,
0.11 mol) in water (15 mL) with efficient stirring at
0e5 ꢂC. Stirring was continued for 20e30 min, and
the resulting fresh cold, flavone-7-diazonium chlo-
Yield: 0.26 g, 33.0%; mp: 200e202 ꢂC. 1H NMR (300 MHz,
CDCl3):
d
1.76 (s, 1H, N(40)eH), 2.46 (s, 3H, CH3), 2.98 (m, 4H, H2-
20 þ H2-60), 3.00 (m, 4H, H2-30 þ H2-50), 6.74 (s, 1H, H-3), 7.11 (dd,
J ¼ 8.7, 1.7 Hz, 1H, H-6), 7.33 (d, J ¼ 1.7 Hz, 1H, H-8), 7.49e7.51 (m,
3H, H-30 þ H-50 þ H-40), 7.88e7.91 (m, 2H, H-20 þ H-60), 8.13 (d,
J ¼ 8.7 Hz, 1H, H-5), 9.37 (s, 1H, NeH). 13C NMR (75 MHz, CDCl3):
d
26.1 (CH3), 46.6 (C-30/C-50), 49.3 (C-20/C-60), 101.1 (C-8), 107.6 (C-
ride/2-methylchromenone-7-diazonium
solution was used immediately as such for the
following coupling reaction.
chloride
3), 112.7 (C-6), 118.5 (C-4a), 126.2 (C-200/C-600), 127.4 (C-400), 129.1 (C-
300/C-500), 131.5 (C-5), 131.9 (C-100), 145.5 (C-7), 147.3 (eC]N), 158.0
(C-8a), 163.0 (C-2), 177.8 (C-4), 195.2 (O]CeMe). HRMS (ESI) m/z:
Calcd for C22H23N4O3 [M þ H]þ 391.17702; found 391.17647. Anal.
Calcd for C22H22N4O3 (390.44 g/mol): C, 67.68, H, 5.68, N, 14.35,
found: C, 67.44, H, 5.58, N, 14.19.
Step (ii). A cold (0e5 ꢂC) freshly prepared solution of 2-methyl-
or 2-phenyl-4-oxo-4H-chromene-7-diazonium chlo-
ride (0.1 mol) was poured onto cold solution
(ꢁ5e0 ꢂC, ice-salt bath) of 3-chloropentane-2,4-dione
(13.5 g, 0.1 mol) in pyridine/water (160 mL, 3:2 v/v)
with vigorous stirring. The resulting orange-colored
mixture was further stirred until a solid precipitate
was formed (5e10 min). The reaction mixture was
then diluted with cold water (200 mL), the solid
product was collected by suction filtration, washed
several times with cold water, dried and recrystallized
from suitable solvent.
4.3.2. 7-{2-[1-(4-methylpiperazin-1-yl)-2-oxopropylidene]
hydrazinyl}-2-phenyl-4H-chromen-4-one (6a)
Yield: 0.14 g, 17.7%; mp: 145e147 ꢂC. 1H NMR (300 MHz,
CDCl3):
d 2.35 (s, 3H, CH3), 2.47 (s, 3H, CH3eN), 2.53 (m, 4H, H2-
30 þ H2-50), 3.10 (m, 4H, H2-20 þ H2-60), 6.76 (s, 1H, H-3), 7.12 (dd,
J ¼ 8.7, 2.0 Hz, 1H, H-6), 7.35 (d, J ¼ 2.0 Hz, 1H, H-8), 7.49 (m, 1H,
H-400), 7.50e7.54 (m, 2H, H-300 þ H-500), 7.90e7.94 (m, 2H, H-
200 þ H-600), 8.15 (d, J ¼ 8.7 Hz, 1H, H-5), 9.26 (s, 1H, NeH). 13C NMR
(75 MHz, CDCl3):
d
26.1 (C]OeCH3), 46.5 (NeCH3), 48.0 (C-20/C-
60), 55.8 (C-30/C-50), 101.0 (C-8), 107.6 (C-3), 112.6 (C-6), 118.5 (C-
4a), 126.3 (C-200/C-600), 127.4 (C-400), 129.1 (C-300/C-500), 131.5 (C-5),
131.9 (C-100), 145.5 (C-7), 147.3 (eC]N), 158.0 (C-8a), 163.1 (C-2),
177.8 (C-4), 195.1 (O]CeMe). HRMS (ESI) m/z: Calcd for
C23H25N4O3 [M þ H]þ 405.19267; found 405.19212. Anal. Calcd for
C23H24N4O3 (404.46 g/mol): C, 68.30, H, 5.98, N, 13.85, found: C,
68.18, H, 5.92, N, 13.76.
4.2.1. 2-oxo-N-(4-oxo-2-phenyl-4H-chromen-7-yl)
propanehydrazonoyl chloride 4a
Yield: 33.0 g, 96.9%; mp: 271e272 ꢂC (recryst. from ethanol). 1H
NMR (300 MHz, CDCl3): d 2.64 (s, 3H, CH3), 6.79 (s,1H, H-3), 7.20 (m,
1H, H-6), 7.39 (d, J ¼ 1.8 Hz, 1H, H-8), 7.52e7.54 (m, 3H, H-30 þ H-
40 þ H-50), 7.90e7.93 (m, 2H, H-20 þ H-60), 8.22 (d, J ¼ 8.5 Hz,1H, H-5),
8.64 (s,1H, NeH).13C NMR (75 MHz, CDCl3):
d 26.1 (CH3),102.6 (C-8),
107.4 (C-3), 113.8 (C-6), 118.8 (C-4a), 126.2 (C-10), 126.8 (C-20/C-60),
127.0 (C-40),129.6 (C-30/C-50),131.7 (C-7),132.2 (C-5),148.0 (eC]N),
157.5 (C-8a), 162.6 (C-2), 176.8 (C-4), 188.7 (O]CeMe). HRMS (ESI)
m/z: Calcd for C18H13ClN2O3Na [M þ Na]þ 363.05124; found
363.05069. Anal. Calcd for C18H13ClN2O3 (340.76 g/mol): C, 63.44, H,
3.85, N, 8.22, found: C, 63.18, H, 3.74, N, 8.04.
4.3.3. 7-{2-[1-(4-ethylpiperazin-1-yl)-2-oxopropylidene]
hydrazinyl}-2-phenyl-4H-chromen-4-one (7a)
Yield: 0.27 g, 32.1%; mp: 165e167 ꢂC. 1H NMR (300 MHz, CDCl3):
d
1.08 (t, J ¼ 7.2 Hz, 3H, CH3eCH2e), 2.38 (s, 3H, C]OeCH3), 2.44 (q,
J ¼ 7.2 Hz, 2H, CH3eCH2eN), 2.52 (m, 4H, H2-30 þ H2-50), 3.08 (m,
4H, H2-20 þ H2-60), 6.69 (s, 1H, H-3), 7.08 (dd, J ¼ 8.7, 2.0 Hz, 1H, H-
6), 7.28 (d, J ¼ 2.0 Hz, 1H, H-8), 7.43e7.47 (m, 3H, H-300 þ H-400 þ H-
500), 7.83e7.87 (m, 2H, H-200 þ H-600), 8.08 (d, J ¼ 8.7 Hz, 1H, H-5),
4.2.2. N-(2-methyl-4-oxo-4H-chromen-7-yl)-2-oxo-propanehydrazo
noyl chloride 4b
9.26 (s, 1H, NeH). 13C NMR (75 MHz, CDCl3):
d 12.0 (CH3eCH2), 26.1
Yield: 24.2 g, 87%; mp:274e276 ꢂC (recryst. from acetonitrile). 1H
(C]OeCH3), 48.0 (NeCH2), 49.5 (C-20/C-60), 52.5 (C-30/C-50), 101.0
(C-8), 107.5 (C-3), 112.7 (C-6), 118.4 (C-4a), 126.2 (C-200/C-600), 127.3
(C-400), 128.5 (C-300/C-500), 131.5 (C-5), 131.8 (C-100), 145.5 (C-7), 147.4
NMR (300 MHz, DMSO-d6):
d 2.38 (s, 3H, CH3-2), 2.40 (s, 3H, O]
CeCH3), 6.11 (s, 1H, H-3), 7.27 (d, J ¼ 1.8 Hz, 1H, H-8), 7.40 (dd,

70
M.N. Abu-Aisheh et al. / European Journal of Medicinal Chemistry 54 (2012) 65e74
(eC]N), 157.9 (C-8a), 163.0 (C-2), 177.7 (C-4), 195.1 (O]CeMe).
HRMS (ESI) m/z: Calcd for C24H27N4O3 [M þ H]þ 419.20832;
found 419.20777. Anal. Calcd for C24H26N4O3 (418.49 g/mol): C,
68.88, H, 6.26, N, 13.39, found: C, 68.64, H, 6.18, N, 13.26.
(MeCH2e),101.6 (C-8),107.4 (C-3),113.6 (C-6),117.7 (C-4a),126.7 (C-
200/C-400/C-600), 129.6 (C-300/C-500), 131.8 (C-100), 132.1 (C-5), 144.8 (C-
7), 148.9 (eC]N), 155.2 (C-8a), 157.7 (O]CeN), 162.5 (C-2), 176.8
(C-4), 195.6 (O]CeMe). HRMS (ESI) m/z: Calcd for C25H27N4O5
[M þ H]þ 463.19814; found 463.19760. Anal. Calcd for C25H26N4O5
(462.50 g/mol): C, 64.92, H, 5.67, N, 12.11, found: C, 64.73, H, 5.54, N,
12.02.
4.3.4. 7-{2-[1-(4-ethylpiperazin-1-yl)-2-oxopropylidene]hydrazinyl}
2-methyl-4H-chromen-4-one (7b)
Yield: 0.23 g, 32.0%; mp: 160e162 ꢂC. 1H NMR (300 MHz, CDCl3):
d
1.07 (t, J ¼ 7.1 Hz, 3H, CH3eCH2e), 2.35 (s, 3H, CH3-2), 2.39 (s, 3H,
4.3.8. Ethyl 4-{1-[2-(2-methyl-4-oxo-4H-chromen-7-yl)hydrazono]-
2-oxopropyl}piperazine-1-carboxylate (9b)
O]CeCH3), 2.44 (q, 2H, J ¼ 7.1 Hz, CH3eCH2eN), 2.53 (m, 4H, H2-
30 þ H2-50), 3.09 (m, 4H, H2-20 þ H2-60), 6.10 (s, 1H, H-3), 7.06 (dd,
J ¼ 8.7 Hz, 2.0 Hz, 1H, H-6), 7.21 (d, J ¼ 2.0 Hz, 1H, H-8), 8.08 (d,
J ¼ 8.7 Hz, 1H, H-5), 9.29 (s, 1H, NeH). 13C NMR (75 MHz, CDCl3):
Yield: 0.60 g, 75.1%; mp: 212e214 ꢂC. 1H NMR (300 MHz, CDCl3):
d
1.26 (t, J ¼ 7.1 Hz, 3H, CH3eCH2), 2.32 (s, 3H, CH3-2), 2.44 (s, 3H,
O]CeCH3), 3.02 (m, 4H, H2-30 þ H2-50), 3.59 (m, 4H, H2-20 þ H2-60),
4.15 (q, J ¼ 7.1 Hz, 2H, MeCH2), 6.06 (d, J ¼ 1.0 Hz, 1H, H-3), 7.07 (dd,
J ¼ 8.8, 2.0 Hz, 1H, H-6), 7.21 (d, J ¼ 2.0 Hz, 1H, H-8), 8.09 (d,
J ¼ 8.8 Hz, 1H, H-5), 9.29 (s, 1H, NeH). 13C NMR (75 MHz, CDCl3):
d
12.1 (CH3eCH2), 20.6 (CH3-2), 26.1 (O]CeCH3), 47.7 (NeCH2),
49.7 (C-20/C-60), 52.8 (C-30/C-50), 100.7 (C-8), 110.5 (C-3), 112.3 (C-
6), 118.3 (C-4a), 127.3 (C-5), 145.7 (C-7), 147.3 (eC]N), 158.1 (C-8a),
165.7 (C-2), 177.8 (C-4), 195.1 (O]CeMe). HRMS (ESI) m/z: Calcd for
C19H25N4O3 [M þ H]þ 357.19267; found 357.19212. Anal. Calcd for
C19H24N4O3 (356.42 g/mol): C, 64.03, H, 6.79, N, 15.72, found: C,
64.12, H, 6.68, N, 15.56.
d
14.7 (CH3CH2e), 20.6 (CH3-2), 26.0 (O]CeCH3), 47.9 (C-20/C-60),
49.9 (C-30/C-50), 61.7 (MeCH2e), 101.0 (C-8), 110.5 (C-3), 112.3 (C-6),
118.3 (C-4a), 127.4 (C-5), 144.7 (C-7), 146.8 (eC]N), 155.5 (O]
CeN), 158.1 (C-8a), 165.8 (C-2), 177.7 (C-4), 195.0 (O]CeMe).
HRMS (ESI) m/z: Calcd for C20H25N4O5 [M þ H]þ 401.18249; found
401.18302. Anal. Calcd for C20H24N4O5 (400.43 g/mol): C, 59.99, H,
6.04, N, 13.99, found: C, 60.12, H, 5.96, N, 13.84.
4.3.5. 7-{2-[1-(4-benzylpiperazin-1-yl)-2-oxopropylidene]
hydrazinyl}-2-phenyl-4H-chromen-4-one (8a)
Yield: 0.87 g, 90.4%; mp: 194e195 ꢂC. 1H NMR (300 MHz, CDCl3):
d
2.47 (s, 3H, CH3), 2.56 (m, 4H, H2-30 þ H2-50), 3.09 (m, 4H, H2-
4.3.9. 7-{2-[1-(4-(2-hydroxyethyl)piperazin-1-yl)-2-oxopropylidene]
hydrazinyl}-2-methyl-4H-chromen-4-one (10b)
20 þ H2-60), 3.57 (s, 2H, NeCH2e), 6.75 (s, 1H, H-3), 7.11 (dd, J ¼ 8.7,
2.0 Hz, 1H, H-6), 7.24e7.34 (m, 5H, H-2000 þ H-3000 þ H-4000 þ H-
5000 þ H-6000), 7.36 (d, J ¼ 2.2 Hz, 1H, H-8), 7.49 (m, 1H, H-400),
7.50e7.52 (m, 2H, H-300 þ H-500), 7.90e7.93 (m, 2H, H-20 þ H-60), 8.14
(d, J ¼ 8.7 Hz, 1H, H-5), 9.28 (s, 1H, NeH). 13C NMR (75 MHz, CDCl3):
Yield: 0.48 g, 64.3%; mp: 147e149 ꢂC. 1H NMR (300 MHz, CDCl3):
d
2.33 (s, 3H, CH3-2), 2.43 (s, 3H, O]CeCH3), 2.61 (t, J ¼ 5.6 Hz, 2H,
eNCH2CH2OH), 2.64 (m, 4H, H2-30 þ H2-50), 3.09 (m, 4H, H2-
20 þ H2-60), 3.63 (t, J ¼ 5.6 Hz, 2H, eNCH2CH2OH), 6.09 (s, 1H, H-3),
7.05 (dd, J ¼ 8.7, 2.0 Hz,1H, H-6), 7.21 (d, J ¼ 2.0 Hz,1H, H-8), 8.08 (d,
J ¼ 8.7 Hz, 1H, H-5), 9.23 (s, 1H, NeH). 13C NMR (75 MHz, CDCl3):
d
26.2 (CH3), 48.0 (C-20/C-60), 53.8 (C-30/C-50), 63.2 (CH2eN), 101.0
(C-8), 107.6 (C-3), 112.7 (C-6), 118.5 (C-4a), 126.3 (C-200/C-600), 127.3
(C-400), 127.4 (C-4000), 128.4 (C-3000/C-5000), 129.1 (C-300/C-500), 129.2 (C-
d
20.6 (CH3-2), 26.0 (O]CeCH3), 48.0 (C-20/C-60), 53.5 (C-30/C-50),
2
000/C-6000), 131.5 (C-5), 131.9(C-100), 137.9 (C-1000), 145.6 (C-7), 147.4
57.8 (eNCH2CH2OH), 59.5 (eNCH2CH2OH), 100.9 (C-8), 110.5 (C-3),
112.3 (C-6), 118.1 (C-4a), 127.3 (C-5), 145.2 (C-7), 147.0 (eC]N),
158.2 (C-8a), 165.7 (C-2), 177.7 (C-4), 195.1 (O]CeMe). HRMS
(ESI) m/z: Calcd for C19H25N4O4 [M þ H]þ 373.18758; found
373.18680. Anal. Calcd for C19H24N4O4 (372.42 g/mol): C, 61.28, H,
6.50, N, 15.04, found: C, 61.06, H, 6.42, N, 14.92.
(eC]N), 158.0 (C-8a), 163.1 (C-2), 177.8 (C-4), 195.2 (O]CeMe).
HRMS (ESI) m/z: Calcd for C29H29N4O3 [M þ H]þ 481.22397;
found 481.22342. Anal. Calcd for C29H28N4O3 (480.56 g/mol): C,
72.48, H, 5.87, N, 11.66, found: C, 72.25, H, 5.74, N, 11.48.
4.3.6. 7-{2-[1-(4-benzylpiperazin-1-yl)-2-oxopropylidene]
hydrazinyl}-2-methyl-4H-chromen-4-one (8b)
4.3.10. 7-[2-(2-oxo-1-(piperidin-1-yl)propylidene)hydrazinyl]-2-
phenyl-4H-chromen-4-one (11a)
Yield: 0.46 g, 54.8%; mp: 190e192 ꢂC. 1H NMR (300 MHz, CDCl3):
d
2.34 (s, 3H, CH3-2), 2.43 (s, 3H, O]CeCH3), 2.54 (m, 4H, H2-
Yield: 0.59 g, 75.7%; mp: 172e173 ꢂC. 1H NMR (300 MHz, CDCl3):
30 þ H2-50), 3.07 (m, 4H, H2-20 þ H2-60), 3.56 (s, 2H, NeCH2e), 6.09
(s, 1H, H-3), 7.05 (dd, J ¼ 8.7 Hz, 1.9 Hz, 1H, H-6), 7.21 (d, J ¼ 1.9 Hz,
1H, H-8), 7.24e7.33 (m, 5H, H-200 þ H-300 þ H-400 þ H-500 þ H-600),
8.08 (d, J ¼ 8.7 Hz, 1H, H-5), 9.23 (s, 1H, NeH). 13C NMR (75 MHz,
d
1.62 (m, 6H, H2-30 þ H2-40 þ H2-50), 2.45 (s, 3H, CH3), 3.00 (m, 4H,
H2-20 þ H2-60), 6.73 (s, 1H, H-3), 7.10 (dd, J ¼ 8.7, 2.0 Hz, 1H, H-6),
7.32 (d, J ¼ 2.0 Hz, 1H, H-8), 7.49e7.51 (m, 3H, H-300 þ H-400 þ H-500),
7.88e7.91 (m, 2H, H-200 þ H-600), 8.12 (d, J ¼ 8.7 Hz, 1H, H-5), 9.28 (s,
CDCl3):
d
20.6 (CH3-2), 26.0 (O]CeCH3), 48.0 (C-20/C-60), 53.8 (C-
1H, NeH). 13C NMR (75 MHz, CDCl3): 24.0 (CH3), 26.2 (C-40), 26.7
d
30/C-50), 63.2 (NeCH2), 100.8 (C-8), 110.5 (C-3), 112.3 (C-6), 118.0 (C-
4a), 127.2 (C-5), 127.3 (C-400), 128.4 (C-200/C-600), 129.1 (C-300/C-500),
137.9 (C-100), 145.5 (C-7), 147.1 (eC]N), 158.2 (C-8a), 165.7 (C-2),
177.7 (C-4), 195.1 (O]CeMe). HRMS (ESI) m/z: Calcd for
C24H27N4O3 [M þ H]þ 419.20832; found 419.20792. Anal. Calcd for
C24H26N4O3 (418.49 g/mol): C, 68.88, H, 6.26, N, 13.39, found: C,
68.72, H, 6.18, N, 13.45.
(C-30/C-50), 49.3 (C-20/C-60), 100.9 (C-8), 107.6 (C-3), 112.6 (C-6),
118.3 (C-4a), 126.2 (C-200/C-600), 127.3 (C-400), 129.0 (C-300/C-500), 131.5
(C-5), 131.9 (C-100), 146.7 (C-7), 147.5 (eC]N), 158.0 (C-8a), 163.0 (C-
2), 177.8 (C-4), 195.4 (O]CeMe). HRMS (ESI) m/z: Calcd for
C23H24N3O3 [M þ H]þ 390.18177; found 390.18122. Anal. Calcd for
C23H23N3O3 (389.45 g/mol): C, 70.93, H, 5.95, N, 10.79, found: C,
70.68, H, 5.86, N, 10.66.
4.3.7. Ethyl 4-[2-oxo-1-(2-(4-oxo-2-phenyl-4H-chromen-7-yl)
hydrazono)propyl] piperazine-1-carboxylate (9a)
4.3.11. 2-methyl-7-{2-[2-oxo-1-(piperidin-1-yl)propylidene]
hydrazinyl}-4H-chromen-4-one (11b)
Yield: 0.59 g, 63.3%; mp: 165e166 ꢂC. 1H NMR (300 MHz, CDCl3):
Yield: 0.56 g, 85.2%; mp: 206e208 ꢂC. 1H NMR (300 MHz,
d
1.17 (t, J ¼ 7.0 Hz, 3H, CH3eCH2), 2.39 (s, 3H, O]CeCH3), 2.91 (m,
CDCl3):
d
1.61 (m, 6H, H2-30 þ H2-40 þ H2-50), 2.33 (s, 3H, CH3-2),
4H, H2-30 þ H2-50), 3.53 (m, 4H, H2-20 þ H2-60), 4.03 (d, J ¼ 7.0 Hz,
2H, MeCH2), 6.91 (s, 1H, H-3), 7.50e7.57 (m, 5H, H-300 þ H-400 þ H-
500 þ H-6 þ H-8), 7.94 (d, J ¼ 8.7 Hz, 1H, H-5), 8.03e8.06 (m, 2H, H-
2.42 (s, 3H, O]CeCH3), 2.95 (m, 4H, H2-20 þ H2-60), 6.07 (s, 1H, H-
3), 7.04 (dd, J ¼ 8.7, 1.9 Hz, 1H, H-6), 7.19 (d, J ¼ 1.9 Hz, 1H, H-8), 8.07
(d, J ¼ 8.7 Hz, 1H, H-5), 9.29 (s, 1H, NeH). 13C NMR (75 MHz, CDCl3):
600 þ H-200), 10.25 (s, 1H, NeH). 13C NMR (75 MHz, CDCl3):
d
15.1
d
20.6 (CH3-2), 24.4 (C-40), 26.0 (O]CeCH3), 26.7 (C-30/C-50), 49.2
(CH3CH2e), 26.6 (O]CeCH3), 44.0 (C-20/C-60), 47.8 (C-30/C-50), 61.3
(C-20/C-60), 100.4 (C-8), 110.4 (C-3), 112.3 (C-6), 117.9 (C-4a), 127.2

M.N. Abu-Aisheh et al. / European Journal of Medicinal Chemistry 54 (2012) 65e74
71
(C-5), 146.6 (C-7), 147.2 (eC]N), 158.2 (C-8a), 165.6 (C-2), 177.7 (C-
4.3.16. 7-{2-[2-oxo-1-(4-phenylpiperazin-1-yl)propylidene]
4), 195.5 (O]CeMe). HRMS (ESI) m/z: Calcd for C18H22N3O3
[M þ H]þ 328.16612; found 328.16557. Anal. Calcd for C18H21N3O3
(327.38 g/mol): C, 66.04, H, 6.47, N,12.84, found: C, 65.87, H, 6.38, N,
12.71.
hydrazinyl}-2-phenyl-4H-chromen-4-one (14a)
Yield: 0.51 g, 54.7%; mp: 212e213 ꢂC. 1H NMR (300 MHz, CDCl3):
2.50 (s, 3H, CH3), 3.18 (m, 4H, H2-30 þ H2-50), 3.24 (m, 4H, H2-
d
20 þ H2-60), 6.73 (s, 1H, H-3), 6.89 (t, J ¼ 7.3 Hz, 1H, H-4000), 6.95 (d,
J ¼ 7.9 Hz, 2H, H-2000 þ H-6000), 7.12 (dd, J ¼ 8.7, 2.0 Hz, 1H, H-6), 7.27
(m, 2H, H-3000 þ H-5000), 7.34 (d, J ¼ 2.0 Hz, 1H, H-8), 7.49e7.51 (m,
3H, H-300 þ H-400 þ H-500), 7.87e7.90 (m, 2H, H-200 þ H-600), 8.13 (d,
J ¼ 8.7 Hz, 1H, H-5), 9.38 (s, 1H, NeH). 13C NMR (75 MHz, CDCl3):
4.3.12. 7-[2-(1-morpholino-2-oxopropylidene)hydrazinyl]-2-phenyl-
4H-chromen-4-one (12a)
Yield: 0.41 g, 51.9%; mp: 209e210 ꢂC. 1H NMR (300 MHz, CDCl3):
d
2.47 (s, 3H, CH3), 3.08 (t, J ¼ 4.5 Hz, 4H, H2-20 þ H2-60), 3.8 (t,
d
26.1 (CH3), 48.2 (C-20/C-60), 50.1 (C-30/C-50), 101.2 (C-8), 107.6 (C-
J ¼ 4.5 Hz, 4H, H2-30 þ H2-50), 6.72 (s, 1H, H-3), 7.13 (dd, J ¼ 8.7,
2.0 Hz, 1H, H-6), 7.34 (d, J ¼ 2.0 Hz, 1H, H-8), 7.46e7.50 (m, 3H, H-
300 þ H-400 þ H-500), 7.87e7.90 (m, 2H, H-200 þ H-600), 8.12 (d,
J ¼ 8.7 Hz, 1H, H-5), 9.41 (s, 1H, NeH). 13C NMR (75 MHz, CDCl3):
3), 112.7 (C-6), 116.5 (C-2000/C-6000), 118.6 (C-4a), 120.3 (C-4000), 126.2
(C-200/C-600), 127.4 (C-400), 129.1 (C-300/C-500), 129.3 (C-3000/C-5000),
131.5 (C-5), 131.7 (C-l00), 145.2 (C-1000), 147.3 (C-7), 151.3 (eC]N),
158.0 (C-8a), 163.0 (C-2), 177.7 (C-4), 195.2 (O]CeMe). HRMS
(ESI) m/z: Calcd for [M þ H]þ 467.20832; found 467.20777. Anal.
Calcd for C28H26N4O3 (466.53 g/mol): C, 72.09, H, 5.62, N, 12.01,
found: C, 71.93, H, 5.51, N, 11.89.
d
26.1 (CH3), 48.3 (C-20/C-60), 67.4 (C-30/C-50), 101.2 (C-8), 107.6 (C-
3), 112.7 (C-6), 118.6 (C-4a), 126.2 (C-200/C-600), 127.4 (C-400), 129.1 (C-
300/C-500), 131.6 (C-5), 131.8 (C-100), 144.7 (C-7), 147.2 (eC]N), 157.9
(C-8a), 163.1 (C-2), 177.7 (C-4), 195.1 (O]CeMe). HRMS (ESI) m/z:
Calcd for C22H20N3O4 [M ꢁ H]ꢁ 390.14538; found 390.14593. Anal.
Calcd for C22H21N3O4 (391.42 g/mol): C, 67.51, H, 5.41, N, 10.74,
found: C, 67.32, H, 5.44, N, 10.63.
4.3.17. 2-methyl-7-{2-[2-oxo-1-(4-phenylpiperazin-1-yl)
propylidene]hydrazinyl}-4H-chromen-4-one (14b)
Yield: 0.52 g, 64.2%; mp: 257e258 ꢂC. 1H NMR (300 MHz,
CDCl3):
d 2.34 (s, 3H, CH3-2), 2.41 (s, 3H, O]CeCH3), 3.24 (m, 4H,
4.3.13. 2-methyl-7-{2-[1-morpholino-2-oxopropylidene]
hydrazinyl}-4H-chromen-4-one (12b)
H2-30 þ H2-50), 3.26 (m, 4H, H2-20 þ H2-60), 6.09 (s, 1H, H-3), 6.89 (t,
J ¼ 7.3 Hz, 1H, H-400), 6.95 (d, J ¼ 8.0 Hz, 2H, H-200 þ H-600), 7.07 (dd,
J ¼ 8.7 Hz, 2.0 Hz, 1H, H-6), 7.21 (d, J ¼ 2.0 Hz, 1H, H-8), 7.27 (m, 2H,
H-300 þ H-500), 8.09 (d, J ¼ 8.7 Hz, 1H, H-5), 9.31 (s, 1H, NeH). 13C
Yield: 0.48 g, 72.6%; mp: 242e244 ꢂC. 1H NMR (300 MHz,
CDCl3):
d 2.35 (s, 3H, CH3-2), 2.45 (s, 3H, O]CeCH3), 3.07 (m, 4H,
H2-20 þ H2-60), 3.79 (m, 4H, H2-30 þ H2-50), 6.10 (s, 1H, H-3), 7.08
(dd, J ¼ 8.6, 1.8 Hz, 1H, H-6), 7.22 (d, J ¼ 1.8 Hz, 1H, H-8), 8.10 (d,
J ¼ 8.6 Hz, 1H, H-5), 9.32 (s, 1H, NeH). 13C NMR (75 MHz, CDCl3):
NMR (75 MHz, CDCl3): d
20.6 (CH3-2), 26.0 (O]CeCH3), 48.2 (C-20/
C-60), 50.2 (C-30/C-50), 100.0 (C-8), 110.5 (C-3), 112.3 (C-6), 118.1 (C-
4a), 120.3 (C-400), 116.5 (C-200/C-600), 127.4 (C-5), 129.2 (C-300/C-500),
145.1 (C-7), 147.0 (eC]N), 151.3 (C-100), 158.2 (C-8a), 161.3 (C-2),
177.7 (C-4), 195.1 (O]CeMe). HRMS (ESI) m/z: Calcd for
C23H25N4O3 [M þ H]þ 405.19267; found 405.19212. Anal. Calcd for
C23H24N4O3 (404.46 g/mol): C, 68.30, H, 5.98, N, 13.85, found: C,
68.12, H, 5.95, N, 13.68.
d
20.6 (CH3-2), 26.0 (O]CeCH3), 48.2 (C-20/C-60), 67.5 (C-30/C-50),
101.0 (C-8), 110.5 (C-3), 112.3 (C-6), 118.0 (C-4a), 127.4 (C-5), 144.6
(C-7), 146.9 (eC]N), 158.2 (C-8a), 165.7 (C-2), 177.6 (C-4), 195.0
(O]CeMe). HRMS (ESI) m/z: Calcd for C17H20N3O4 [M þ H]þ
330.14538; found 330.14483. Anal. Calcd for C17H19N3O4 (329.35 g/
mol): C, 62.00, H, 5.81, N, 12.76, found: C, 61.84, H, 5.82, N, 12.65.
4.3.18. 7-(2-{1-[4-(4-fluorophenyl)piperazin-1-yl]-2-oxopropylidene}
hydrazinyl)-2-phenyl-4H-chromen-4-one (15a)
4.3.14. 7-[2-(2-oxo-1-thiomorpholinopropylidene)hydrazinyl]-2-
phenyl-4H-chromen-4-one (13a)
Yield: 0.25 g, 25.3%; mp: 225e227 ꢂC. 1H NMR (300 MHz,
Yield: 0.53 g, 65.6%; mp: 217e218 ꢂC. 1H NMR (300 MHz, CDCl3):
CDCl3):
d
2.50 (s, 3H, CH3), 3.22 (m, 4H, H2-30 þ H2-50), 3.25 (m, 4H,
d
2.46 (s, 3H, CH3), 2.75 (m, 4H, H2-30 þ H2-50), 3.27 (m, 4H, H2-
H2-20 þ H2-60), 6.75 (s, 1H, H-3), 6.88e6.93 (m, 2H, H-2000 þ H-6000),
6.94e7.00 (m, 2H, H-3000 þ H-5000), 7.13 (dd, J ¼ 8.7, 2.0 Hz, 1H, H-6),
7.35 (d, J ¼ 2.0 Hz, 1H, H-8), 7.48e7.52 (m, 3H, H-300 þ H-400 þ H-500),
7.88e7.92 (m, 2H, H-200 þ H-600), 8.14 (d, J ¼ 8.7 Hz, 1H, H-5), 9.36 (s,
20 þ H2-60), 6.74 (s, 1H, H-3), 7.13 (dd, J ¼ 8.7, 2.0 Hz, 1H, H-6), 7.33
(d, J ¼ 2.0 Hz, 1H, H-8), 7.47e7.51 (m, 3H, H-300 þ H-400 þ H-500),
7.88e7.91 (m, 2H, H-200 þ H-600), 8.13 (d, J ¼ 8.7 Hz, 1H, H-5), 9.24 (s,
1H, NeH). 13C NMR (75 MHz, CDCl3):
d
26.0 (CH3), 28.5 (C-30/C-50),
1H, NeH). 13C NMR (75 MHz, CDCl3): 26.1 (CH3), 48.2 (C-20/C-60),
d
50.4 (C-20/C-60), 101.2 (C-8), 107.6 (C-3), 112.7 (C-6), 118.6 (C-4a),
126.2 (C-200/C-600), 127.4 (C-400), 129.0 (C-300/C-500), 131.5 (C-5), 131.8
(C-100), 145.8 (C-7), 147.2 (eC]N), 157.9 (C-8a), 163.1 (C-2), 177.7
(C-4), 195.1 (O]CeMe). HRMS (ESI) m/z: Calcd for C22H22N3O3S
[M þ H]þ 408.13819; found 408.13764. Anal. Calcd for C22H21N3O3S
(407.49 g/mol): C, 64.85, H, 5.19, N, 10.31, found: C, 64.68, H, 5.12, N,
10.22.
51.1 (C-30/C-50), 101.2 (C-8), 107.6 (C-3), 112.6 (C-6), 115.4 (d,
2JCeF ¼ 22.0 Hz, C-3000/C-5000), 118.3 (d, JCeF ¼ 7.6 Hz, C-2000/C-6000),
3
118.6 (C-4a), 126.2 (C-200/C-600), 127.4 (C-400), 129.1 (C-300/C-500), 131.6
(C-5), 131.9 (C-100), 145.2 (C-7), 147.3 (eC]N), 147.9 (d,
4JCeF ¼ 2.3 Hz, C-1000), 157.5 (d, 1JCeF ¼ 240 Hz, C-4000), 158.0 (C-8a),
163.1 (C-2), 177.7 (C-4), 195.2 (O]CeMe). HRMS (ESI) m/z: Calcd for
C28H26FN4O3 [M þ H]þ 485.19442; found 485.19535. Anal. Calcd for
C28H25FN4O3 (484.52 g/mol): C, 69.41, H, 5.20, N, 11.56, found: C,
69.18, H, 5.23, N, 11.46.
4.3.15. 2-methyl-7-{2-[2-oxo-1-thiomorpholinopropylidene]
hydrazinyl}-4H-chromen-4-one (13b)
Yield: 0.44 g, 63.4%; mp: 195e197 ꢂC. 1H NMR (300 MHz, CDCl3):
4.3.19. 7-{2-[1-(4-(4-fluorophenyl)piperazin-1-yl) -2-oxopropylidene]
hydrazinyl}-2-methyl-4H-chromen-4-one (15b)
d
2.34 (s, 3H, CH3-2), 2.43 (s, 3H, O]CeCH3), 2.74 (m, 4H, H2-30 þ H2-
50), 3.26 (m, 4H, H2-20 þ H2-60), 6.10 (s, 1H, H-3), 7.07 (dd, J ¼ 8.5,
Yield: 0.58 g, 68.5%; mp: 218e220 ꢂC. 1H NMR (300 MHz,
1.8 Hz,1H, H-6), 7.20 (d, J ¼ 1.8 Hz,1H, H-8), 8.08 (d, J ¼ 8.5 Hz,1H, H-
CDCl3):
d 2.34 (s, 3H, CH3-2), 2.47 (s, 3H, O]CeCH3), 3.20 (m, 4H,
5), 9.17 (s, 1H, NeH). 13C NMR (75 MHz, CDCl3):
d
20.6 (CH3-2), 25.9
H2-30 þ H2-50), 3.22 (m, 4H, H2-20 þ H2-60), 6.09 (s, 1H, H-3),
6.88e7.00 (m, 4H, H-200 þ H-300 þ H-500 þ H-600), 7.07 (dd, J ¼ 8.7 Hz,
1.9 Hz, 1H, H-6), 7.21 (d, J ¼ 1.9 Hz, 1H, H-8), 8.09 (d, J ¼ 8.7 Hz, 1H,
(O]CeCH3), 28.5 (C-30/C-50), 50.4 (C-20/C-60),101.0 (C-8),110.5 (C-3),
112.3 (C-6),118.0 (C-4a),127.4 (C-5),145.7 (C-7),146.9 (eC]N),158.2
(C-8a), 165.8 (C-2), 177.7 (C-4), 195.1 (O]CeMe). HRMS (ESI) m/z:
Calcd for C17H20N3O3S [M þ H]þ 346.12254; found 346.12199. Anal.
Calcd for C17H19N3O3S (345.42 g/mol): C, 59.11, H, 5.54, N, 12.17,
found: C, 59.18, H, 5.51, N, 12.06.
H-5), 9.27 (s, 1H, NeH). 13C NMR (75 MHz, CDCl3):
d 20.6 (CH3-4),
26.0 (O]CeCH3), 48.2 (C-20/C-60), 51.1 (C-30/C-50), 101.0 (C-8), 110.5
(C-3), 112.3 (C-6), 115.7 (d, 2JCeF ¼ 22.5 Hz, C-300/C-500), 118.1 (C-4a),
3
118.3 (d, JCeF ¼ 7.5 Hz, C-200/C-600), 127.4 (C-5), 145.0 (C-7), 147.0

72
M.N. Abu-Aisheh et al. / European Journal of Medicinal Chemistry 54 (2012) 65e74
(eC]N), 147.7 (C-100), 157.0 (d, 1JCeF ¼ 240 Hz, C-400), 158.2 (C-8a),
165.7 (C-2),177.6 (C-4),195.2 (O]CeMe). HRMS (ESI) m/z: Calcd for
C23H24FN4O5 [M þ H]þ 423.18324; found 423.18270. Anal. Calcd for
C23H23FN4O3 (422.45 g/mol): C, 65.39, H, 5.49, N, 13.26, found: C,
65.13, H, 5.41, N, 13.08.
J ¼ 4.7 Hz, 1H, H-500), 7.09 (dd, J ¼ 8.7, 2.0 Hz, 1H, H-6), 7.23 (d,
J ¼ 2.0 Hz, 1H, H-8), 8.09 (d, J ¼ 8.7 Hz, 1H, H-5), 8.32 (d, J ¼ 4.7 Hz,
2H, H-400 þ H-600), 9.39 (s, 1H, NeH). 13C NMR (75 MHz, CDCl3):
d
20.6 (CH3-2), 26.0 (O]CeCH3), 44.3 (C-20/C-60), 48.0 (C-30/C-50),
101.0 (C-8), 110.4 (C-500), 110.5 (C-3), 112.3 (C-6), 118.2 (C-4a), 127.4
(C-5), 145.0 (C-7), 146.9 (eC]N), 157.8 (C-400/C-600), 158.2 (C-8a),
161.6 (C-200), 165.7 (C-2), 177.6 (C-4), 195.2 (O]C-Me). HRMS (ESI)
m/z: Calcd for C21H23N6O3 [M þ H]þ 407.18316; found 407.18276.
Anal. Calcd for C21H22N6O3 (406.44 g/mol): C, 62.06, H, 5.46, N,
20.68, found: C, 61.88, H, 5.40, N, 20.54.
4.3.20. 7-(2-{1-[4-(2-fluorophenyl)piperazin-1-yl]-2-oxopropylidene}
hydrazinyl)-2-phenyl-4H-chromen-4-one (16a)
Yield: 0.28 g, 29.1%; mp: 225e227 ꢂC. 1H NMR (300 MHz,
CDCl3):
d
2.50 (s, 3H, CH3), 3.22 (m, 4H, H2-30 þ H2-50), 3.23 (m, 4H,
H2-20 þ H2-60), 6.76 (s, 1H, H-3), 6.96e7.08 (m, 4H, H-3000 þ H-
4000 þ H-5000 þ H-6000), 7.14 (dd, J ¼ 8.7, 2.0 Hz, 1H, H-6), 7.37 (d,
J ¼ 2.0 Hz, 1H, H-8), 7.50e7.53 (m, 3H, H-300 þ H-400 þ H-500),
7.90e7.93 (m, 2H, H-200 þ H-600), 8.15 (d, J ¼ 8.7 Hz, 1H, H-5), 9.36 (s,
4.3.24. 7-(2-{1-[4-(4-methoxyphenyl)piperazin-1-yl]-2-
oxopropylidene}hydrazinyl)-2-phenyl-4H-chromen-4-one (18a)
Yield: 0.31 g, 31.6%; mp: 207e208 ꢂC. 1H NMR (300 MHz, CDCl3):
1H, NeH). 13C NMR (75 MHz, CDCl3):
d
26.1 (CH3), 48.3 (C-20/C-60),
d
2.50 (s, 3H, CH3), 3.18 (m, 4H, H2-30 þ H2-50), 3.24 (m, 4H, H2-20 þ H2-
51.4 (C-30/C-50), 101.2 (C-8), 107.6 (C-3), 112.7 (C-6), 116.4 (d,
2JCeF ¼ 20.6 Hz, C-3000), 118.6 (C-4a), 119.2 (d, 4JCeF ¼ 2.9 Hz, C-5000),
123.0 (d, 3JCeF ¼ 7.9 Hz, C-4000), 124.5 (d, 3JCeF ¼ 3.5 Hz, C-6000), 126.3
(C-200/C-600), 127.4 (C-400), 129.1 (C-300/C-500), 131.6 (C-5), 131.9 (C-100),
139.5 (d, 2JCeF ¼ 8.6 Hz, C-1000), 145.3 (C-7), 147.3 (eC]N), 155.5 (d,
1JCeF ¼ 244.6 Hz, C-2000), 158.0 (C-8a), 163.1 (C-2), 177.8 (C-4), 195.1
(O]CeMe). HRMS (ESI) m/z: Calcd for C28H26FN4O3 [M þ H]þ
485.19442; found 485.19435. Anal. Calcd for C28H25FN4O3
(484.52 g/mol): C, 69.41, H, 5.20, N, 11.56, found: C, 69.15, H, 5.14, N,
11.42.
60), 3.76 (s, 3H, OCH3), 6.75 (s,1H, H-3), 6.85 (m, 2H, H-2000 þ H-6000), 6.93
(m, 2H, H-3000 þ H-5000), 7.12 (dd, J ¼ 8.7, 2.0 Hz, 1H, H-6), 7.35 (d,
J ¼ 2.0 Hz, 1H, H-8), 7.48e7.52 (m, 3H, H-300 þ H-400 þ H-500), 7.89e7.92
(m, 2H, H-200 þ H-600), 8.14 (d, J ¼ 8.7 Hz,1H, H-5), 9.36 (s,1H, NeH). 13C
NMR (75 MHz, CDCl3):
d
26.8 (CH3), 48.0 (C-20/C-60), 50.1 (C-30/C-50),
55.7 (OCH3), 101.5 (C-8), 107.4 (C-3), 113.6 (C-6), 114.8 (C-3000/C-5000),
117.6 (C-4a), 118.1 (C-2000/C-6000), 126.7 (C-200/C-600), 128.8 (C-400), 129.6
(C-300/C-500),131.1 (C-100),132.1 (C-5),145.3 (C-1000),146.2 (C-7),149.0 (C-
4000), 153.4 (eC]N), 157.7 (C-8a), 162.5 (C-2), 176.8 (C-4), 195.6 (O]
CeMe). HRMS (ESI) m/z: Calcd for C29H29N4O4 [M þ H]þ 497.21888;
found 497.21833. Anal. Calcd for C29H28N4O4 (496.56 g/mol): C, 70.15,
H, 5.68, N, 11.28, found: C, 69.94, H, 5.56, N, 11.21.
4.3.21. 7-{2-[1-(4-(2-fluorophenyl)piperazin-1-yl)-2-oxopropylidene]
hydrazinyl}-2-methyl-4H-chromen-4-one (16b)
Yield: 0.56 g, 65.9%; mp: 195e197 ꢂC. 1H NMR (300 MHz, CDCl3):
4.3.25. 7-(2-(1-(4-(4-methoxyphenyl)piperazin-1-yl)-2-
oxopropylidene)hydrazinyl)-2-methyl-4H-chromen-4-one (18b)
Yield: 0.76 g, 87.1%; mp: 254e256 ꢂC. 1H NMR (300 MHz, CDCl3):
d
2.34 (s, 3H, CH3-2), 2.47 (s, 3H, O]CeCH3), 3.20 (m, 4H, H2-
30 þ H2-50), 3.23 (m, 4H, H2-20 þ H2-60), 6.09 (s, 1H, H-3), 6.96e7.07
(m, 5H, H-6 þ H-300 þ H-400 þ H-500 þ H-600), 7.20 (m, 1H, H-8), 8.08
(d, J ¼ 8.0 Hz, 1H, H-5), 9.31 (s, 1H, NeH). 13C NMR (75 MHz, CDCl3):
d
2.34 (s, 3H, CH3-2), 2.47 (s, 3H, O]CeCH3), 3.17 (m, 4H, H2-30 þ H2-50),
3.22 (m, 4H, H2-20 þ H2-60), 3.76 (s, 3H, OeCH3), 6.08 (s, 1H, H-3), 6.84
(d, J ¼ 9.0 Hz, H-200 þ H-600), 6.92 (d, J ¼ 9.0 Hz, H-300 þ H-500), 7.06 (dd,
J¼ 8.7 Hz,1.9 Hz,1H, H-6), 7.21 (d, J¼ 1.9 Hz,1H, H-8), 8.08 (d, J¼ 8.7 Hz,
d
20.6 (CH3-2), 26.0 (O]CeCH3), 48.2 (C-20/C-60), 51.3 (C-30/C-50),
101.0 (C-8), 110.5 (C-3), 112.3 (C-6), 118.1 (C-4a), 116.2 (d,
4
2JCeF ¼ 21.0 Hz, C-300), 119.2 (d, JCeF ¼ 2.8 Hz, C-500), 122.9 (d,
1H, H-5), 9.30 (s, 1H, NeH). 13C NMR (75 MHz, CDCl3):
d 20.5 (CH3-2),
3JCeF ¼ 8.3 Hz, C-400),124.6 (d, 3JCeF ¼ 3.4 Hz, C-600),127.3 (C-5),140.0
26.0 (O]CeCH3), 48.3 (C-20/C-60), 51.6 (C-30/C-50), 55.6 (OCH3), 100.9
(C-8), 110.4 (C-3), 112.3 (C-6), 114.5 (C-300/C-500), 118.2 (C-4a), 118.6 (C-
200/C-600),127.3 (C-5),154.2 (C-400), 145.7 (C-100),145.2 (C-7),147.0 (eC]
N), 158.3 (C-8a), 177.6 (C-4), 177.6 (C-4), 195.1 (O]CeMe). HRMS (ESI)
m/z:CalcdforC24H27N4O4 [M þ H]þ 435.19541; found 435.20068. Anal.
Calcd for C24H26N4O4 (434.49 g/mol): C, 66.34, H, 6.03, N,12.89, found:
C, 66.15, H, 5.95, N, 12.82.
2
(d, JCeF ¼ 9.0 Hz, C-100), 145.1 (C-7), 147.0 (eC]N), 158.2 (C-8a),
155.8 (d, 1JCeF ¼ 245.0 Hz, C-200), 165.8 (C-2), 177.7 (C-4), 195.1 (O]
CeMe). HRMS (ESI) m/z: Calcd for C23H22FN4O3 [M
421.16759; found 421.16705. Anal. Calcd for C23H23FN4O3 (422.45 g/
mol): C, 65.39, H, 5.49, N, 13.26, found: C, 65.23, H, 5.46, N, 13.12.
ꢁ
H]ꢁ
4.3.22. 7-{2-[2-oxo-1-(4-(pyrimidin-2-yl)piperazin-1-yl)propylidene]
hydrazinyl}-2-phenyl-4H-chromen-4-one (17a)
4.3.26. 7-(2-{1-[4-(4-chlorophenyl)piperazin-1-yl]-2-oxopropylidene}
hydrazinyl)-2-phenyl-4H-chromen-4-one (19a)
Yield: 0.64 g, 68.2%; mp: 202e203 ꢂC. 1H NMR (300 MHz,
CDCl3):
d
2.48 (s, 3H, CH3), 3.13 (m, 4H, H2-30 þ H2-50), 3.94 (m, 4H,
Yield: 0.29 g, 28.8%; mp: 204e205 ꢂC. 1H NMR (300 MHz, DMSO-
H2-20 þ H2-60), 6.51 (t, J ¼ 4.7 Hz,1H, H-5000), 6.76 (s,1H, H-3), 7.15 (d,
J ¼ 8.6 Hz, 1H, H-6), 7.37 (s, 1H, H-8), 7.50e7.52 (m, 3H, H-300 þ H-
400 þ H-500), 7.90e7.92 (m, 2H, H-200 þ H-600), 8.15 (d, J ¼ 8.6 Hz, 1H,
H-5), 8.32 (d, J ¼ 4.7 Hz, 2H, H-4000 þ H-6000), 9.45 (s, 1H, NeH). 13C
d6):
d
2.41 (s, 3H, CH3), 3.13 (m, 4H, H2-30 þ H2-50), 3.27 (m, 4H, H2-
20 þ H2-60), 6.90 (s, 1H, H-3), 6.97 (d, J ¼ 9.0 Hz, 2H, H-2000 þ H-6000),
7.23 (d, J ¼ 9.0, 2H, H-3000 þ H-5000), 7.51 (dd, J ¼ 8.7, 2.0 Hz, 1H, H-6),
7.53e7.59 (m, 4H, H-300 þ H-400 þ H-500 þ H-8), 7.94 (d, J ¼ 8.7 Hz,1H,
H-5), 8.05 (m, 2H, H-200 þ H-600),10.2 (s,1H, NeH). 13C NMR (75 MHz,
NMR (75 MHz, CDCl3):
d
26.7 (CH3), 43.9 (C-20/C-60), 47.7 (C-30 þ C-
50), 101.6 (C-8), 107.3 (C-3), 110.6 (C-5000), 113.7 (C-6), 117.7 (C-4a),
126.7 (C-200/C-400/C-600), 129.7 (C-300/C-500), 131.8 (C-100), 132.1 (C-5),
145.1 (C-7), 149.0 (eC]N), 157.7 (C-8a), 158.5 (C-4000/C-6000), 161.8
(C-2000), 162.5 (C-2), 176.8 (C-4), 195.7 (O]CeMe). HRMS (ESI) m/z:
Calcd for C26H25N6O3 [M þ H]þ 469.19881; found 469.19827. Anal.
Calcd for C26H24N6O3 (468.51 g/mol): C, 66.65, H, 5.16, N, 17.94,
found: C, 66.43, H, 5.06, N, 17.72.
DMSO-d6): d
26.7 (CH3), 47.7 (C-20/C-60), 48.6 (C-30/C-50),101.6 (C-8),
107.4 (C-3), 113.6 (C-6), 117.5 (C-2000/C-6000), 117.7 (C-4a),122.8 (C-3000
/
C-5000), 126.7 (C-200/C-600), 129.2 (C-400), 129.7 (C-300/C-500), 131.9 (C-
100), 132.1 (C-5), 141.4 (C-4000), 145.1 (C-1000), 149.0 (C-7), 150.5 (eC]
N), 157.7 (C-8a), 162.6 (C-2), 176.8 (C-4), 195.6 (O]CeMe). HRMS
(ESI) m/z: Calcd for C28H24ClN4O3 [M ꢁ H]ꢁ 499.15369; found
499.15424. Anal. Calcd for C28H25ClN4O3 (500.98 g/mol): C, 67.13, H,
5.03, N, 11.18, found: C, 67.04, H, 5.12, N, 11.06.
4.3.23. 2-methyl-7-(2-(2-oxo-1-(4-(pyrimidin-2-yl)piperazin-1-yl)
propylidene)hydrazinyl)-4H-chromen-4-one (17b)
4.4. Cell lines and cell culture
Yield: 0.58 g, 71.2%; mp: 232e235 ꢂC. 1H NMR (300 MHz, CDCl3):
d
2.34 (s, 3H, CH3-2), 2.44 (s, 3H, O]CeCH3), 3.11 (m, 4H, H2-
The K562 leukemia cell line was obtained from Dr. Mona Has-
sona (Faculty of Science, The University of Jordan) and was cultured
30 þ H2-50), 3.92 (m, 4H, H2-20 þ H2-60), 6.08 (s, 1H, H-3), 6.50 (t,

M.N. Abu-Aisheh et al. / European Journal of Medicinal Chemistry 54 (2012) 65e74
73
(b) M. Weidenborner, H. Hindrof, H.C. Jha, P. Tsotsonos, Antifungal activity of
flavonoids against storage fungi of the genus Aspergillus, Phytochemistry 29
(1990) 1103e1105;
(c) M. Weidenborner, H.C. Jha, Antifungal activity of flavonoids and their
mixtures against different fungi occurring on grain, Science 38 (1993)
347e351.
in RPMI while the T47D and MCF-7 breast cancer cells were
obtained from American Type culture collections (ATCC) and were
cultured in DMEM/F12. All media were supplemented with 10%
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m
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