Efficient microwave-assisted synthesis and antitumor activity of novel 4,4′-methylenebis[2-(3-aryl-4,5-dihydro-1H-pyrazol-5-yl)phenols]

Article abstract of DOI:10.1016/j.ejmech.2011.03.028

Two new series of 4,4′-methylenebis[2-(3-aryl-4,5-dihydro-1H-pyrazol- 5-yl)phenols] 4 and methylenebis-2-(3-aryl-4,5-dihydro-1H-pyrazol-5-yl)-4,1- phenylene diacetates 5 have been efficiently prepared starting from the commercially available salicylaldehy

Full text of DOI:10.1016/j.ejmech.2011.03.028

European Journal of Medicinal Chemistry 46 (2011) 2436e2440
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Efficient microwave-assisted synthesis and antitumor activity of novel
4,40-methylenebis[2-(3-aryl-4,5-dihydro-1H-pyrazol-5-yl)phenols]
,
a
a
a
a
Braulio Insuasty a , Angélica García , Jairo Quiroga , Rodrigo Abonia , Alejandro Ortiz ,
*
Manuel Nogueras b, Justo Cobo b
a Heterocyclic Compounds Research Group, Department of Chemistry, Universidad del Valle, A.A. 25360 Cali, Colombia
b Department of Inorganic and Organic Chemistry, Universidad de Jaén, 23071 Jaén, Spain
a r t i c l e i n f o
a b s t r a c t
Two new series of 4,40-methylenebis[2-(3-aryl-4,5-dihydro-1H-pyrazol-5-yl)phenols] 4 and methyl-
enebis-2-(3-aryl-4,5-dihydro-1H-pyrazol-5-yl)-4,1-phenylene diacetates 5 have been efficiently prepa-
red starting from the commercially available salicylaldehyde and where the key step corresponded to the
microwave-assisted reaction of intermediates 3,30-[methylenebis(6-hydroxy-3,1-phenylene)]bis(1-aryl-
prop-2-en-1-ones) 3 with hydrazine hydrate or acetic acid/hydrazine hydrate respectively. Three unac-
etylated compounds type 4 presented interesting antitumor activities against a wide range of tumor cell
lines.
Article history:
Received 12 November 2010
Received in revised form
4 March 2011
Accepted 14 March 2011
Available online 23 March 2011
Keywords:
Bis-diarylpyrazolines
Ó 2011 Elsevier Masson SAS. All rights reserved.
Microwave irradiation
Antitumor activity
Hydrazine hydrate
3,30-[Methylenebis(6-hydroxy-3,1-
phenylene)]bis(1-arylprop-2-en-1-ones)
1. Introduction
hypotension [8] and inflammatory diseases [9]. Furthermore,
compounds bearing two pyrazolinic rings have also shown signif-
Pyrazoline derivates are five-member nitrogenated heterocyclic
compounds associated with a broad spectrum of biological activi-
ties. Several pyrazoline derivatives have been investigated as
inhibitors of mammalian monoamine oxidase (MAO)-A and-B [1]
and antitumor agents. Baraldi et al. have synthesized and evalu-
ated a series of hybrids of pyrazole analogues for in vitro cytotoxic
activity against a variety of cancer cell lines [2], Antonini et al.
reported the synthesis, antitumor cytotoxicity, and DNA-binding of
icant pharmacological activities [10e12].
On the other hand, the development of high-throughput
methodologies for drug discovery is highly demanded, and con-
cretely the microwave-assisted reactions have gained considerable
importance because of its simplicity, increasing reaction rates and
cleaner product formation; including its well adaptation to a wide
variety of experimental variants, such as solvent free approaches
[13e21].
novel N-5,2-di(
dine-5-carboxamides using (
u
-aminoalkyl)-2,6-dihydropyrazolo[3,4,5-kl]acri-
-aminoalkyl)hydrazines as starting
In this paper, we are reporting the synthesis of two series of 4,40-
methylenebis[2-(3-aryl-4,5-dihydro-1H-pyrazol-5-yl)phenols]
4aee and methylenebis-2-(3-aryl-4,5-dihydro-1H-pyrazol-5-yl)-
4,1-phenylene diacetates 5aee whose synthesis has been planned
from the commercially available salicylaldehyde via the formation
of key intermediates 3,30-[methylenebis(6-hydroxy-3,1-phenyl-
ene)]bis(1-arylprop-2-en-1-ones) 3aee. The further microwave-
assisted reaction with hydrazine hydrate, or with a mixture of
hydrazine hydrate and acetic acid, respectively, afforded the
desired products. Several of these compounds were tested for
antitumor activity against 60 different human tumor cell lines,
where products 4a, 4c and 4e showed interesting values of GI50
u
materials [3], Hernandez et al. recently were reported the synthesis
of a novel pyrazolodibenzo[1,4]diazepines by a tandem sequence
amine-exchange/heterocyclization starting from readily available
enaminones and arylhydrazines, some of such derivatives shown
a important antitumor activity [4]. Additionally, pyrazole deriva-
tives have presented antibacterial activity [5] and pharmaceu-
tical properties for using in the treatment of depression [6,7],
* Corresponding author. Tel.: þ57 2 3393248; fax: þ57 2 3392440.
E-mail address: braulio.insuasty@correounivalle.edu.co (B. Insuasty).
and LC50
.
0223-5234/$ e see front matter Ó 2011 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejmech.2011.03.028

B. Insuasty et al. / European Journal of Medicinal Chemistry 46 (2011) 2436e2440
2437
2. Results and discussion
2.1. Chemistry
2.2. Anticancer activity
Three (4a, 4c and 4e) of the ten compounds proposed to NCI
(National Cancer Institute of the United States) for screening were
selected, and made the preliminary evaluation against the 60
In order to obtain the key
a,b-unsaturated intermediates as
starting materials for the synthesis of the target products 4, 5,50-
methylene-bis-salicylaldehyde 2 was obtained from the commer-
cially available salicylaldehyde, which then underwent reaction
with substituted acetophenones leading to the formation of the
methylene-bis-chalcones 3aee [22,23]. Treatment of chalcones 3,
with hydrazine hydrate under microwave irradiation, at times no
longer than 2 min, allowed the formation of the desired 4,40-
methylenebis[2-(3-aryl-4,5-dihydro-1H-pyrazol-5-yl)phenols]
4aee in good to excellent yields. The addition of acetic acid to the
above mixture under the same reaction conditions afforded the
methylenebis-2-(3-aryl-4,5-dihydro-1H-pyrazol-5-yl)-4,1-phenyl-
ene diacetates 5aee, in just one step and in also good yields
(Scheme 1).
The new series of compounds 4 and 5 were fully characterized
by means of spectroscopic techniques such as FT-IR, 1D-, and 2D-
NMR and EI-MS, as summarized in Experimental Section. The FT-IR
spectra for compounds 4aee showed the characteristic absorption
bands for OH and NH stretching at 3470e3451 cmꢀ1 and
3418e3329 cmꢀ1 respectively. As expected, compounds 5aee did
not show the absorption band for the OH group, and did for the NH.
The main feature to be remarked from the 1H NMR spectra of
compounds 4aee and 5aee, corresponded to typical signals of the
pyrazoline ring. For example, both structures showed an AMX
coupling system, involving the diastereotopic H-4 protons and the
H-5 proton as showed in Experimental Section. All mass spectra of
series 4 and 5 exhibited well-defined molecular ion peaks.
tumor cell lines at a single dose of 1.0 mM. The output from the
single dose screening was reported as a mean graph available for
analysis by the COMPARE program. The results of the first assay
showed that compounds 4a and 4c were active while compound 4e
was inactive.
Then, active compounds passed to a second stage in order to
determine their cytostatic activity, they were evaluated at five
concentration levels against various tumor cell lines like melanoma,
leukemia and cancers of lung, colon, brain, breast, ovary, kidney or
prostate. The test consisted of a 48 h continuous drug exposure
protocol using sulforhodamide B (SRB) protein assay to estimate cell
growth. Details of this evaluation method and the complementary
information related with the activity pattern over all cell lines have
been published [24e26]. Finally, the compound 4a exhibited activity
against 60 human tumor cell lines, with low values of GI50 w 10ꢀ6 M
in all cell lines, i.e. GI50 ¼ 9.48 ꢁ 10ꢀ6 M for SK-MEL-28 (Melanoma),
or GI50 ¼ 1.75 ꢁ10ꢀ6 M for K-562 (Leukemia), and in general inter-
esting LC50 values, higher than 1 ꢁ10ꢀ4 M. In a similar way,
compound 4c also showed an interesting activity against 32 human
tumorcell lines, remarking thatfor SK-MEL-5 (Melanoma) with GI50
:
1.76 ꢁ 10ꢀ6 M and LC50: 3.17 ꢁ 10ꢀ5 M. Consequently, compounds 4a
and 4c exhibited significant activities, with GI50 ranges from 10ꢀ6 to
10ꢀ5 M. The cytotoxicities associated with the latter compounds,
measured as LC50 are around 100 mM, for most cell lines, indicating
a low toxicity of such compounds for normal human cell lines as
required for potential antitumor agents (see Table 1).
HO
OH
OH
Trioxane
H2SO4
OHC
CHO
CHO
1
2
O
R
KOH/EtOH
OH
HO
c) CH3
d) OCH3
b) Br e) 3,4-OCH2O-
R =
a) Cl
R
R
O
O
3a-e
NH2NH2.H2O
HOAc
NH2NH2.H2O
MW
MW
6
m
6
m
AcO
OAc
4
5
5
4
1
o
i
p
HO
NH
OH
4
1
2
5
o
i
p
4
5
2
3
3
3
R
R
3
N
NH
HN
N
2
R
R
N
HN
N
2
1
1
5a-e
4a-e
R
Cl
Br
CH3
OCH3
3,4-OCH3O-
Comp.
Yield(%)
mp (ºC)
4a
5a
95
4b
5b
97
4c
5c
4d
5d
4e
5e
98
98
89
87
97
89
90
83
260-262 164-166 265-267 194-196 267-269 145-147 250-252 156-158 279-281 179-180
1.5 1.0 1.5 1.0 1.5 1.0 2.0 1.5 2.0 1.5
t.r. (min)
Scheme 1. Synthesis of 4,40-methylene-bis-2-(3-aryl-4,5-dihydro-1H-pyrazol-5-yl)phenols 4aef and 4,40-methylene-bis-2-(3-aryl-4,5-dihydro-1H-pyrazol-5-yl-4,1-phenylene)
diacetate 5aef.

2438
B. Insuasty et al. / European Journal of Medicinal Chemistry 46 (2011) 2436e2440
Table 1 (continued)
Table 1
In vitro testing expressed as growth inhibition of cancer cell lines for compounds 4a
Panel/cell line
Compounds
and 4c.a
4a
4c
Panel/cell line
Compounds
b
c
b
c
GI50
(
mM)
LC50
(
m
M)
GI50
(
mM)
LC50 (mM)
4a
4c
BT-549
T-47D
MDA-MB-468
4.78
3.79
3.50
>100
>100
>100
4.66
4.61
2.97
>100
>100
>100
b
c
b
c
GI50
(
m
M)
LC50
(m
M)
GI50
(
m
M)
LC50 (mM)
Leukemia
CCRF-CEM
HL-60(TB)
K-562
MOLT-4
RPMI-8226
a
1.94
2.66
1.75
2.70
5.09
>100
>100
>100
>100
>100
3.85
e
>100
e
Data obtained from NCI’s in vitro disease-oriented human tumor cell lines screen
[27].
b
7.73
e
>100
e
GI50 was the drug concentration resulting in a 50% reduction in the net protein
increase (as measured by SRB staining) in control cells during the drug incubation.
Determined at five concentration levels (100, 10, 1.0, 0.1 and 0.01 M).
9.89
>100
m
c
LC50 is a parameter of cytotoxicity and reflects the molar concentration needed
Non-small cell lung
SR
A549/ATCC
EKVX
HOP-62
HOP-92
NCI-H226
NCI-H23
NCI-H322M
NCI-H460
NCI-H522
to kill 50% of the cells.
1.83
2.45
2.77
4.16
3.39
4.74
3.30
4.17
2.15
2.95
>100
>100
>100
>100
>100
>100
>100
>100
>100
e
e
6.84
e
>100
e
8.64
8.28
8.28
5.30
e
>100
>100
>100
>100
e
3. Conclusion
We have performed the synthesis of two novel series of bis-
diarylpyrazoline derivates in a synthetic sequence starting from
the commercially available salicylaldehyde, with a final key step
which involved an efficient microwave-assisted cyclocondensation
between methylene-bis-chalcones 3 and hydrazine in short reac-
tion times and excellent yields. Biological assays on compounds 4a
and 4c exhibited promising activities against different tumor cell
lines with remarkable values of GI50: 1.75e9.48
their LC50 values (>100 M). As compound 4a showed an impor-
tant activity against K-562 of Leukemia, it will be taken as a leader
molecule for the development of new series of bispyrazoline
derivates looking for improving the antitumor activity of this
family of compounds.
4.88
3.64
>100
>100
46.2
Colon
COLO 205
HCC-2998
HCT-116
HCT-15
HT29
5.44
4.88
1.85
2.51
7.25
3.09
3.29
>100
e
e
e
>100
>100
>100
>100
>100
>100
e
4.11
3.64
e
55.2
>100
mM, along with
e
m
KM12
SW-620
e
e
7.96
>100
CNS
SF-268
SF-295
SF-539
SNB-19
SNB-75
U251
3.22
1.47
5.82
3.64
4.26
1.80
>100
>100
>100
>100
>100
>100
e
e
e
e
e
e
6.89
4.98
5.84
>100
>100
>100
4. Experimental
Commercially available starting materials, reagents and solvents
were used as supplied. TLC analyses were performed on Merck
silica gel 60 F254 aluminum plates. Melting points were determined
in a Buchi Melting Point Apparatus and are uncorrected. The 1H and
13C NMR spectra were run on a Bruker AVANCE 400 spectrometer
operating at 400 MHz and 100 MHz respectively, using dime-
thylsulfoxide-d6 as solvent and tetramethylsilane as internal
reference. The mass spectra were scanned on a Shimadzu GCMS-QP
2010 spectrometer (equipped with a direct inlet probe) and oper-
ating at 70 eV. The elemental analyses have been obtained using
a Thermo Finnigan Flash EA1112 CHN (STIUJA) elemental analyzer.
Microwave experiments were carried out on a focused microwave
reactor (300 W CEM DiscoverÔ).
Melanoma
LOX IMVI
MALME-3M
M14
MDA-MB-435
SK-MEL-2
SK-MEL-28
SK-MEL-5
UACC-257
UACC-62
1.82
5.08
4.45
3.21
3.23
9.84
3.36
2.56
3.16
>100
>100
>100
>100
85.5
>100
36.7
>100
43.6
7.38
e
>100
e
e
e
8.52
5.90
e
>100
>100
e
1.76
e
31.7
e
e
e
Ovarian
IGROV1
4.11
2.59
3.54
6.10
2.46
2.24
4.43
>100
>100
>100
>100
>100
>100
>100
e
e
OVCAR-3
OVCAR-4
OVCAR-5
OVCAR-8
NCI/ADR-RES
SK-OV-3
e
e
5.36
e
>100
e
6.36
7.39
e
>100
>100
e
4.1. General procedure for the synthesis of 4,40-methylenebis[2-(3-
aryl-4,5-dihydro-1H-pyrazol-5-yl)phenols] (4aee)
Renal
786-0
A498
ACHN
CAKI-1
RXF 393
SN12C
TK-10
UO-31
A mixture of 0.20 mmol of 3.30-(5,50-methylene-bis-2-hyd-
roxyphenyl)-bis-1-arylprop-2-en-1-ones 3aee and 0.48 mmol of
hydrazine hydrate was subjected to microwave irradiation in
a closed vessel at 150 ꢂC with a maximum power of 300 W for
times no longer than two minutes. The reactions were followed by
thin layer chromatography; the resulting mixture was cooled to
room temperature and then water was added. The precipitate
formed was filtered and washed several times with hot water and
hexane.
3.81
5.37
3.63
2.67
3.99
2.86
5.34
3.04
>100
>100
>100
>100
>100
>100
>100
>100
8.39
e
>100
e
5.56
e
>100
e
4.18
8.84
e
>100
>100
e
4.29
>100
Prostate
PC-3
DU-145
3.32
3.46
>100
>100
e
e
e
e
4.1.1. 4,40-Methylenebis[2-(3-(4-chlorophenyl)-4,5-dihydro-1H-
pyrazol-5-yl)phenol] (4a)
Breast
MCF7
MDA-MB-231/ATCC
HS 578T
3.04
2.65
2.76
>100
>100
>100
3.30
e
71.8
e
This compound was obtained as light brown solid, yield 98%,
5.58
>100
M.p. 260e262 ꢂC, IR (KBr):
y
¼ 3470 for eOH, 3418 for eNH, 3020,

B. Insuasty et al. / European Journal of Medicinal Chemistry 46 (2011) 2436e2440
2439
2980, 1650, 1587 cmꢀ1. 1H NMR (400 MHz, DMSO-d6):
J2 ¼1.3 Hz, Jgem ¼ 15.8 Hz, 2H), 3.36 (dd, J3 ¼ 10.4 Hz, Jgem ¼ 15.8 Hz,
2H), 3.66 (s, 2H), 4.94 (dd, J2 ¼ 1.3 Hz, J3 ¼ 10.4, 2H), 6.67e7.59
(m, 14H), 7.11e7.18 (bs, 2H, OH), 9.34 (s, 2H, NH) ppm; 13C NMR
d
¼ 2.65 (dd,
4.2. General procedure for the synthesis of methylenebis-2-(3-aryl-
4,5-dihydro-1H-pyrazol-5-yl)-4,1-phenylene diacetates 5aee
A mixture of 0.20 mmol of 3.30-(5,50-methylene-bis-2-hydrox-
yphenyl)-bis-1-arylprop-2-en-1-ones 3aee, 0.48 mmol of hydra-
zine hydrate and 0.40 mmol of acetic acid was subjected to
microwave irradiation in a closed vessel at 180 ꢂC with a maximum
power of 300 W for times no longer than 2 min. The reactions were
followed by thin layer chromatography; the resulting mixture was
cooled to room temperature and then water was added. The
precipitate formed was filtered and washed twice with hot water
and hexane.
(DMSO-d6, 100 MHz):
d
¼ 39.5, 40.2, 59.3, 115.4, 127.4, 128.4, 128.5,
128.5, 128.9, 132.4, 132.7, 132.8, 148.6, 153.3 ppm; EI-MS (m/z, %):
560 [M þ 4] (2), 558 [M þ 2] (13), 556 [Mþ] (20), 376 (13), 198 (100),
178 (67). Anal. Calcd. for C31H26Cl2N4O2: C, 66.79; H, 4.70; N, 19.49.
Found: C, 66.69; H, 4.77; N, 19.43.
4.1.2. 4.40-Methylenebis[2-(3-(4-bromophenyl)-4,5-dihydro-1H-
pyrazol-5-yl)phenol] (4b)
This compound was obtained as light brown solid, yield 98%,
M.p. 265e267 ꢂC, IR (KBr):
y
¼ 3451 for eOH, 3329 for eNH, 3017,
4.2.1. Methylenebis-2-(3-(4-chlorophenyl)-4,5-dihydro-1H-
pyrazol-5-yl)-4,1-phenylene diacetates (5a)
2980, 1657, 1586 cmꢀ1. 1H NMR (400 MHz, DMSO-d6):
d
¼ 2.65 (dd,
J2 ¼1.1 Hz, Jgem ¼ 15.6 Hz, 2H), 3.38 (dd, J3 ¼ 10.4 Hz, Jgem ¼ 15.6 Hz,
2H), 3.66 (s, 2H), 4.91 (dd, J2 ¼1.1 Hz, J3 ¼ 10.4 Hz, 2H), 6.68e7.52
(m, 14H), 7.10e7.17 (bs, 2H, OH), 9.36 (s, 2H, NH) ppm; 13C NMR
This compound was obtained as light brown solid, yield 95%,
M.p. 164e166 ꢂC, IR (KBr):
y
¼ 3387 for eNH, 3015, 2983, 1696 for
C]O, 1670, 1523 cmꢀ1
.
1H NMR (400 MHz, DMSO-d6):
d
¼ 2.23
(DMSO-d6, 100 MHz):
d
¼ 39.4, 40.4, 59.3, 115.4, 121.4, 127.3, 128.4,
(s, 6H), 2.95 (dd, J2 ¼1.5 Hz, Jgem ¼ 14.3 Hz, 2H), 3.60 (s, 2H), 3.70
(dd, J3 ¼ 10.3 Hz, Jgem ¼ 14.3 Hz, 2H), 5.53 (dd, J2 ¼1.5 Hz,
J3 ¼ 10.3 Hz, 2H), 6.62e7.74 (m, 14H), 9.40 (s, 2H, NH) ppm; 13C
128.5, 131.5, 131.8, 132.4, 133.0, 148.6, 153.3 ppm; EI-MS (m/z, %):
648 [M þ 4] (6), 646 [M þ 2] (10), 644 [Mþ] (5), 420 (19), 222 (100),
198 (53). Anal. Calcd. for C31H26Br2N4O2: C, 57.60; H, 4.05; N, 8.67.
Found: C, 57.67; H, 4.12; N, 8.61.
NMR (DMSO-d6, 100 MHz):
d
¼ 22.1, 40.0, 40.5, 41.2, 115.8, 126.6,
126.9, 127.8, 128.5, 128.7, 129.2, 130.1, 135.1, 152.5, 154.0, 167.8 ppm;
EI-MS (m/z, %): 644 [M þ 4](3), 642 [M þ 2](12), 640 [Mþ] (16), 580
(31), 280 (100). Anal. Calcd. for C35H30Cl2N4O4: C, 65.53; H, 4.71; N,
8.73. Found: C, 65.59; H, 4.72; N, 8.66.
4.1.3. 4.40-Methylenebis[2-(3-(4-methylphenyl)-4,5-dihydro-1H-
pyrazol-5-yl)phenol] (4c)
This compound was obtained as light brown solid, yield 89%,
M.p. 267e269 ꢂC, IR (KBr):
y
¼ 3456 for eOH, 3330 for eNH, 3024,
4.2.2. Methylenebis-2-(3-(4-bromophenyl)-4,5-dihydro-1H-
pyrazol-5-yl)-4,1-phenylene diacetates (5b)
2979, 1658, 1510 cmꢀ1
.
1H NMR (400 MHz, DMSO-d6):
d
¼ 2.28
(s, 6H), 2.65 (dd, J2 ¼ 2.0 Hz, Jgem ¼ 16.2 Hz, 2H), 3.36 (dd,
J3 ¼10.8 Hz, Jgem ¼ 16.2 Hz, 2H), 3.67 (s, 2H), 4.91 (dd, J2 ¼ 2.0 Hz,
J3 ¼10.8 Hz, 2H), 6.67e7.49 (m, 14H), 7.19e7.25 (bs, 2H, OH), 9.36
This compound was obtained as light brown solid, yield 97%,
M.p. 194e196 ꢂC, IR (KBr):
y
¼ 3380 for eNH, 3025, 2982, 1696 for
C]O, 1674, 1528 cmꢀ1
.
1H NMR (400 MHz, DMSO-d6):
d
¼ 2.23
(s, 2H, NH) ppm; 13C NMR (DMSO-d6, 100 MHz):
d
¼ 21.3, 39.8, 39.8,
(s, 6H), 2.95 (dd, J2 ¼1.0 Hz, Jgem ¼ 16.3 Hz, 2H), 3.60 (s, 2H), 3.71
(m, J3 ¼ 11.5 Hz, Jgem ¼ 16.3 Hz, 2H), 5.53 (dd, J2 ¼1.0 Hz,
J3 ¼ 11.5 Hz, 2H), 6.62e7.65 (m,14H), 9.41 (s, 2H, NH) ppm; 13C NMR
59.3, 115.5, 125.9, 127.5, 128.3, 128.5, 129.5, 131.0, 132.4, 138.0, 150.4,
153.4 ppm; EI-MS (m/z, %): 516 [Mþ] (22), 356 (12), 198 (53), 158
(100). Anal. Calcd. for C34H32N4O2: C, 76.72; H, 6.24; N, 10.84.
Found: C, 76.66; H, 6.19; N, 10.76.
(DMSO-d6, 100 MHz):
d
¼ 22.1, 40.2, 40.4, 41.2, 115.8, 126.6, 126.8,
127.7, 128.5, 128.9, 129.0, 131.1, 135.0, 152.5, 154.1, 167.8 ppm; EI-MS
(m/z, %): 732 [M þ 4] (26), 730 [M þ 2] (47), 728 [Mþ] (24), 668
(100), 280 (71). Anal. Calcd. for C35H30Br2N4O4: C, 57.55; H, 4.14; N,
7.67. Found: C, 57.61; H, 4.12; N, 7.60.
4.1.4. 4,40-Methylenebis[2-(3-(4-methoxyphenyl)-4,5-dihydro-1H-
pyrazol-5-yl)phenol] (4d)
This compound was obtained as light brown solid, yield 97%,
M.p. 250e252 ꢂC, IR (KBr):
y
¼ 3463 for eOH, 3352 for eNH, 3008,
4.2.3. Methylenebis-2-(3-(4-methylphenyl)-4,5-dihydro-1H-
pyrazol-5-yl)-4,1-phenylene diacetates (5c)
2978, 1656, 1550 cmꢀ1. 1H NMR (400 MHz, DMSO-d6):
d
¼ 2.64 (dd,
J2 ¼1.1 Hz, Jgem ¼ 16.4 Hz, 2H), 3.36 (dd, J3 ¼11.0 Hz, Jgem ¼ 16.4 Hz,
2H), 3.68 (s, 2H), 3.75 (s, 6H), 4.89 (dd, J2 ¼1.1 Hz, J3 ¼ 11.0 Hz, 2H),
6.67e7.54 (m,14H), 7.11e7.18 (bs, 2H, OH), 9.38 (s, 2H, NH) ppm; 13C
This compound was obtained as light brown solid, yield 87%,
M.p. 145e147 ꢂC, IR (KBr):
y
¼ 3384 for eNH, 3002, 2989, 1699 for
C]O, 1664, 1539 cmꢀ1
.
1H NMR (400 MHz, DMSO-d6):
d
¼ 2.23
NMR (DMSO-d6, 100 MHz):
d
¼ 40.0, 40.3, 55.6, 59.3, 114.4, 115.5,
(s, 6H), 2.31 (s, 6H), 2.93 (dd, J2 ¼ 1.1 Hz, Jgem ¼ 15.8 Hz, 2H), 3.59
(s, 2H), 3.68 (dd, J3 ¼ 10.7 Hz, Jgem ¼ 15.8 Hz, 2H), 5.50 (dd,
J2 ¼ 1.1 Hz, J3 ¼10.7 Hz, 2H), 6.61e7.62 (m, 14H), 9.39 (s, 2H, NH)
126.4, 127.4, 127.5, 128.3, 128.4, 128.5, 132.4, 153.4, 159.9 ppm;
EI-MS (m/z, %): 548 [Mþ] (32), 372 (100), 198 (49), 174 (60). Anal.
Calcd. for C33H32N4O4: C, 72.24; H, 5.88; N,10.21. Found: C, 72.30; H,
5.94; N, 10.17.
ppm; 13C NMR (DMSO-d6, 100 MHz):
d
¼ 22.0, 22.1, 40.0, 40.3, 41.4,
115.5, 126.5, 126.9, 127.9, 128.0, 128.4, 129.7, 132.1, 140.3, 152.5,
154.9,167.6 ppm; EI-MS (m/z, %): 600 [Mþ] (41), 540 (100), 280 (31).
Anal. Calcd. for C37H36N4O4: C, 73.98; H, 6.04; N, 9.33. Found: C,
73.91; H, 6.12; N, 9.39.
4.1.5. 4,40-Methylenebis[2-(3-(1,3-benzodioxol-5-yl)-4,5-dihydro-
1H-pyrazol-5-yl)phenol] (4e)
This compound was obtained as light brown solid, yield 90%,
M.p. 279e281 ꢂC, IR (KBr):
y
¼ 3470 for eOH, 3359 for eNH, 3006,
4.2.4. Methylenebis-2-(3-(4-methoxyphenyl)-4,5-dihydro-1H-
pyrazol-5-yl)-4,1-phenylene diacetates (5d)
2978, 1667, 1550 cmꢀ1. 1H NMR (400 MHz, DMSO-d6):
d
¼ 2.65 (dd,
J2 ¼ 0.9 Hz, Jgem ¼ 16.3 Hz, 2H), 3.36 (dd, J3 ¼ 10.8 Hz, Jgem ¼ 16.3 Hz,
2H), 3.67 (s, 2H), 4.93 (dd, J2 ¼ 0.9 Hz, J3 ¼10.8 Hz, 2H), 6.06 (s, 4H),
6.67e7.59 (m, 14H), 7.12e7.19 (bs, 2H, OH), 9.39 (s, 2H, NH) ppm;
This compound was obtained as light brown solid, yield 89%, M.p.
156e158 ꢂC, IR (KBr):
y
¼ 3312 for eNH, 3009, 2980, 1698 for C]O,
1663, 1514 cmꢀ1.1H NMR (400 MHz, DMSO-d6):
d
¼ 2.22 (s, 6H), 2.92
13C NMR (DMSO-d6, 100 MHz):
d
¼ 39.5, 40.3, 59.3, 101.2, 115.4,
(dd, J2 ¼ 1.2 Hz, Jgem ¼ 16.8 Hz, 2H), 3.59 (s, 2H), 3.77 (dd, J3 ¼11.0 Hz,
Jgem ¼ 16.8 Hz, 2H), 3.85 (s, 6H), 5.51 (dd, J2 ¼ 1.2 Hz, J3 ¼11.0 Hz, 2H),
6.60e7.79 (m, 14H), 9.39 (s, 2H, NH) ppm; 13C NMR (DMSO-d6,
115.8, 117.3, 127.4, 127.6, 127.9, 128.5, 128.9, 129.9, 153.3, 160.0 ppm;
EI-MS (m/z, %): 576 [Mþ] (23), 198 (47), 188 (100). Anal. Calcd. for
C33H28N4O6: C, 68.74; H, 4.89; N, 9.72. Found: C, 68.67; H, 4.82;
N, 9.68.
100 MHz):
d
¼ 22.0, 40.0, 40.2, 41.5, 46.4, 114.6, 115.8, 121.9, 126.3,
128.4, 128.6, 137.1, 138.1, 145.7, 151.7, 162.9, 169.0 ppm; EI-MS (m/z,

2440
B. Insuasty et al. / European Journal of Medicinal Chemistry 46 (2011) 2436e2440
%): 632 [Mþ] (5), 572 (8), 280 (100). Anal. Calcd. for C37H36N4O6: C,
70.24; H, 5.74; N, 8.86. Found: C, 70.18; H, 5.82; N, 8.79.
[4] S. Hernandez, I. Moreno, R. SanMartin, M.T. Herrero, E. Dominguez, Org.
Biomol. Chem. (2011). doi:10.1039/C0OB00812E.
[5] B.S. Holla, P.M. Akbarali, M.K. Shivanada, Il Farmaco 55 (2000) 256e263.
[6] Y.R. Prassad, A.L. Rao, L. Prasoona, K. Murali, P.R. Kumar, Bioorg. Med. Chem.
Lett. 15 (2005) 5030e5034.
4.2.5. Methylenebis-2-[3-(1,3-benzodioxol-5-yl)-4,5-dihydro-1H-
pyrazol-5-yl]-4,1-phenylene diacetate (5e)
ꢀ
ꢀ
[7] N. Gökhan-Kelekçi, S. Koyunoglu, S. Yabanoglu, K. Yelekçi, Ö. Özgen, G. Uçar,
K. Erol, E. Kendi, A. Yes¸ ilada, Bioorg. Med. Chem. 17 (2009) 675e689.
[8] G. Turan-Zitouni, P. Chevallet, F.S. Kiliç, K. Erol, Eur. J. Med. Chem. 35 (2000)
635e641.
This compound was obtained as light brown solid, yield 83%, M.p.
179e180 ꢂC, IR (KBr):
y
¼ 3310 for eNH, 3000, 2967, 1694 for C]O,
1662,1573 cmꢀ1.1H NMR (400 MHz, DMSO-d6):
d
¼ 2.25 (s, 6H), 2.95
[9] O. Bruno, A. Ranise, F. Bondavalli, F. Schenone, M. D’Amico, A. Filipelli,
W. Filipelli, F. Rossi, Farmaco 48 (1993) 949e966.
[10] A.R. Bhat, F. Athar, A. Azam, Eur. J. Med. Chem. 44 (2009) 426e431.
[11] B. Insuasty, H. Martinez, J. Quiroga, R. Abonía, M. Nogueras, J. Cobo,
J. Heterocycl. Chem. 45 (2008) 1521e1524.
[12] R.P. Ganorkar, V.S. Jamode, Asian J. Chem. 15 (2003) 475e478.
[13] A. Loupy, A. Petit, J. Hamelin, F. Texier-Boullet, P. Jacquault, D. Mathe,
Synthesis (1998) 1213e1234.
(dd, J2 ¼1.1 Hz, Jgem ¼ 16.2 Hz, 2H), 3.62 (s, 2H), 3.72 (dd, J3 ¼11.3 Hz,
Jgem ¼ 16.2 Hz, 2H), 5.54 (dd, J2 ¼ 1.1 Hz, J3 ¼ 11.3 Hz, 2H), 6.08 (s, 4H),
6.67e7.33 (m, 12H), 9.40 (s, 2H, NH) ppm; 13C NMR (DMSO-d6,
100 MHz):
d
¼ 22.1, 39.7, 40.1, 41.5, 101.6, 115.8, 121.6, 126.5, 128.0,
128.5, 130.1, 132.1, 148.2, 149.4, 152.5, 154.7, 167.6 ppm; EI-MS (m/z,
%): 660 [Mþ] (23), 600 (38), 280 (100) m/z. Anal. Calcd. for
C37H32N4O8:C,67.26;H, 4.88;N,8.48. Found:C,67.19;H, 4.80;N,8.55.
[14] R.S. Varma, Green Chem. (1999) 43e55.
[15] S. Deshayes, M. Liagre, A. Loupy, J.-L. Luche, A. Petit, Tetrahedron 55 (1999)
10851e10870.
[16] K. Tanaka, F. Toda, Chem. Rev. 100 (2000) 1025e1074.
[17] R.S. Varma, Clean Prod. Process. 1 (1999) 132e147.
[18] R.S. Varma, Pure Appl. Chem. (2001) 193e198.
Acknowledgements
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275e282.
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Tetrahedron 42 (2001) 5625e5627.
[21] J. Quiroga, A. Rengifo, B. Insuasty, R. Abonía, M. Nogueras, A. Sánchez, Tetra-
hedron Lett. 43 (2002) 9061e9063.
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M. Mayo, M. Boyd, J. Natl. Cancer Inst. 83 (1991) 757e766.
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K.W. Kohn, T. Fojo, S.E. Bates, L.V. Rubinstein, N.L. Anderson, J.K. Buolanwini,
W.W. van Osdol, A.P. Monks, D.A. Scudiero, E.A. Sausiville, D.W. Zaharevitz,
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The authors wish to credit The Developmental Therapeutics
Program (DTP) of the National Cancer Institute of the United States
(U.S.) for performing the screening of compounds. Authors also
thank to Colciencias, Universidad del Valle (Colombia), Consejería
Economía, Innovación y Ciencia (Junta de Andalucía) and Uni-
versidad de Jaén (Spain) for financial support.
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