Synthesis and anticancer activity of isatin-based pyrazolines and thiazolidines conjugates

Article abstract of DOI:10.1002/ardp.201100055

The synthesis and antitumor activity screening of novel isatin based conjugates with thiazolidine and pyrazoline moieties were performed. Reaction of 3,5-diaryl-4,5-dihydropyrazoles with chloroacetyl chloride yielded starting 2-chloro-1-(3,5-diaryl-4,5-di

Full text of DOI:10.1002/ardp.201100055

514
Arch. Pharm. Chem. Life Sci. 2011, 344, 514–522
Full Paper
Synthesis and Anticancer Activity of Isatin-Based
Pyrazolines and Thiazolidines Conjugates
Dmytro Havrylyuk1, Natalya Kovach1, Borys Zimenkovsky1, Olexandr Vasylenko2,
and Roman Lesyk1
1 Department of Pharmaceutical, Organic and Bioorganic Chemistry, Danylo Halytsky Lviv National Medical
University, Ukraine, Lviv, Ukraine
2 Institute of Bioorganic Chemistry and Petrochemistry, National Academy of Science of Ukraine, Kyiv,
Ukraine
The synthesis and antitumor activity screening of novel isatin based conjugates with thiazolidine and
pyrazoline moieties were performed. Reaction of 3,5-diaryl-4,5-dihydropyrazoles with chloroacetyl
chloride yielded starting 2-chloro-1-(3,5-diaryl-4,5-dihydropyrazol-1-yl)-ethanones which were utilized
in alkylation of isatin and 5-bromoisatin. Thus, corresponding 1-[2-(3,5-diaryl-4,5-dihydropyrazol-1-yl)-
2-oxoethyl]-1H-indole-2,3-diones (1a–1d) have been obtained. The compounds 1a–1d have been used in
Knoevenagel condensation with 4-thiazolidinones for obtaining a series of 5-ylidenederivatives 2a–2f
and 3a–3d. The synthesized compounds were tested for their anticancer activity in NCI60 cell
lines. Among the tested compounds, 5-bromo-1-{2-[5-(4-chlorophenyl)-3-(4-methoxyphenyl)-4,5-
dihydropyrazol-1-yl]-2-oxoethyl}-1H-indole-2,3-dione (1d) was found to be the most active candidate
with selective influence on leukemia subpanel tumor cell lines with GI50 values range of 0.69–
3.35 mM.
Keywords: Anticancer activity / Isatines / Pyrazolines / 4-Thiazolidinones
Received: February 14, 2011; Revised: March 21, 2011; Accepted: April 6, 2011
DOI 10.1002/ardp.201100055
Introduction
tic effects [6–8]. Thus, among isatin based conjugates with
benzothiazole [8], thiazolidinone [9, 10] or 4-piperazinylqui-
noline [11] the promising anticancer agents were identified.
On the other hand diazoles (pyrazoles and pyrazolines)
have occupied a unique position in the design and synthesis
of novel biologically active agents that exert remarkable
anticancer activities [12, 13]. The various antitumor diazole
derivatives were identified as inhibitors of cyclin-dependent
kinase [14], heat shock proteins [15], vascular endothelium
growth factors [16], and P-glycoprotein [17]. These obser-
vations have prompted us to synthesized new isatin-pyrazo-
line hybrids with the hope of discovering active and selective
compounds that would elicit synergistic anticancer activity.
The further conjugation of isatin-pyrazoline hybrids with 4-
thiazolidinones have been motivated by the significant
potential of thiazolidine derivatives as antitumor agents [6].
The previous systematic study of 4-thiazolidinone derivatives
with heterocyclic fragments in the molecules allowed us to
identify a number of highly-active antitumor compounds
in vitro [7, 18–21].
The 1H-indole-2,3-dione [1] is the privileged scaffold in the
modern medicinal chemistry which have a broad spectrum
of the biological activity and the wide possibility to the
chemical modification. The evaluation of antitumor activity
is actual and promising for isatin derivatives [2]. The mech-
anisms of antitumor 1H-indole-2,3-diones can be associated
with their affinity to tyrosine kinase [3], cyclin-dependent
kinases (CDKs) [4] and carbonic anhydrase isozymes (CAIs) [5].
It is known that the combination of different bioactive frag-
ments with complementary pharmacophoric functions or
with different mechanisms of action often showed synergis-
Correspondence: Dr. Roman Lesyk, Department of Pharmaceutical,
Organic and Bioorganic Chemistry, Danylo Halytsky Lviv National Medical
University, Ukraine, Pekarska 69, Lviv, 79010, Ukraine.
E-mail: dr_r_lesyk@org.lviv.net
Fax: þ38 0322 75-77-34
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Arch. Pharm. Chem. Life Sci. 2011, 344, 514–522
Isatin-based Pyrazolines and Thiazolidines Conjugates
515
The present work is an extension of our ongoing efforts
towards developing promising antitumor agents by a hybride
pharmacophore approach. We synthesized hybrid com-
pounds by linking the main structural unit of the isatin ring
system with the pyrazoline and 4-thiazolidinone, and
examined their antitumor activity in vitro.
with isatin and 5-bromoisatin in DMF at room temperature
[24]. Thus the corresponding 1-[2-(3,5-diaryl-4,5-dihydropyra-
zol-1-yl)-2-oxoethyl]-1H-indole-2,3-diones 1a–1d have been
obtained (Scheme 1). It is known that nature of ylidene
moiety in position 5 of 4-thiazolidinone cycle has an essential
influence on the antitumor activity [6, 7, 20, 25]. Relying on
these observations and considering the high isatin reactivity
as carbonile compound, we utilized 1-[2-(3,5-diaryl-4,5-dihy-
dropyrazol-1-yl)-2-oxoethyl]-1H-indole-2,3-diones 1a–1d in the
reactions with 2,4-thiazolidinedione, 2-thioxo-4-thiazolidi-
none, and 2-amino-4-thiazolone according to the standard
Knoevenagel condensation procedure (medium – acetic acid,
catalyst – fused sodium acetate) [6, 7, 20, 25, 26]. Following
the mentioned reactions the new non-condensed isatin con-
jugates with pyrazoline and 4-thiazolidinone 2a–2f and 3a–
3d have been synthesized.
Results and discussion
Chemistry
The general methods for synthesis of target isatin based
conjugates with pyrazoline and 4-thiazolidinone moieties
are depicted in Scheme 1.
The starting 3,5-diaryl-4,5-dihydropyrazoles, that were syn-
thesized using known methods from appropriate chalcones
[22] easily reacted with chloroacetyl chloride yielding 2-
chloro-1-(3,5-diaryl-4,5-dihydropyrazol-1-yl)-ethanones [23].
Appropriate 2-chloro-1-(3,5-diaryl-4,5-dihydropyrazol-1-yl)-
ethanones were tested as alkylating agents in the reaction
The characterization data of synthesized novel heterocy-
clic substituted thiazolidones are presented in experimental
part. Analytical and spectral data (IR, 1H-, 13C-NMR) confirmed
the structure of the synthesized compounds.
R2
R2
R3
R2
O
O
R3
O
N
O
O
ClCH2COCl
N
H
NH2NH2
O
N
N
N
NH
a
O
N
N
R1
Cl
R2
R1
1a R1 = OMe, R2 = R3 = H;
1b R1 = Cl, R2 = R3 = H;
R1
R1
O
H
N
O
1c R1 = Cl, R2 = OMe, R3 = H;
1d R1 = Cl, R2 = OMe, R3 = Br
N
X
S
b
NH2
b
S
R3
R2
R3
R2
O
O
N
N
N
O
NH
X
O
N
S
O
N
N
N
S
O
NH2
2a R1 = OMe, R2 = R3 = H, X = S;
2b R1 = Cl, R2 = R3 = H, X = S;
R1
3a R1 = OMe, R2 = R3 = H;
3b R1 = Cl, R2 = R3 = H;
2c R1 = Cl, R2 = OMe, R3 = H, X = S;
R1
2d R1 = Cl, R2 = OMe, R3 = Br, X = S;
2e R1 = Cl, R2 = OMe, R3 = H, X = O;
2f R1 = Cl, R2 = OMe, R3 = Br, X = O
3c R1 = Cl, R2 = OMe, R3 = H;
3d R1 = Cl, R2 = OMe, R3 = Br
Scheme 1. Synthesis of 1,3-dihydroindol-2-one conjugates with 3,5-diaryl-4,5-dihydropyrazole and 4-thiazolidinone moieties.
Reagents, conditions, and yields: (a) K2CO3, DMF, r.t. 12 h, 68–78%; (b) AcONa, AcOH, reflux 2–3 h, 67–79%.
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1
Protons CH2-CH of pyrazoline fragment in the H-NMR spec-
tra of synthesized compounds showed characteristic patterns
of an AMX system. The chemical shifts of the protons HA, HM,
which was described elsewhere (see e.g. http://dtp.nci.nih.
gov) [29–31]. The results of primary screening are reported
as the percent cancer cell line growth (GP%) and presented in
Table 1. The range of growth % shows the lowest and the
highest growth % found among different cancer cell lines.
The most active compound 1d was found to be effective
against 26 cell lines and three compounds (1a, 2e, 3a)
were found to be moderately effective against few cell lines,
while five compounds (1b, 2d, 2f, 3c, 3d) were found to be
ineffective (Table 1).
and HX have been assigned to about
d ꢀ 3.14–3.25, d ꢀ
3.87–3.95, and d ꢀ 5.53–5.61, respectively with corresponding
coupling constants of JAM ¼ 16.8–18.3, JAX ¼ 10.8–11.8, and
JMX ¼ 3.2–4.8 Hz. The protons of the methylene group
(CH2CO) appear as two doublets at
d ꢀ 4.92–
1
5.09 ppm and d ꢀ 5.05–5.17 ppm. In the H-NMR spectra of
2a–2f NH proton of thiazolidinone cycle shows the broad
singlet at d ꢀ 8.82–9.01 and the NH2 protons of 3a–3d have
Finally, compound 1d possessed considerable activity
against most of tested human tumor cell lines and was
selected in advanced assay against a panel of approximately
sixty tumor cell lines at 10-fold dilutions of five concen-
trations (100 mM, 10 mM, 1 mM, 0.1 mM, and 0.01 mM) (see
also http://dtp.nci.nih.gov) [29–31]. Based on the cytotoxicity
assays, three antitumor activity dose-response parameters
were calculated for experimental agents against each cell
line: GI50 – molar concentration of the compound that
inhibits 50% net cell growth; TGI – molar concentration of
the compound leading to total inhibition; and LC50 – molar
concentration of the compound leading to 50% net cell
death. Furthermore a mean graph midpoints (MG_MID) were
calculated for each of the parameters, giving an average
activity parameter over all cell lines for tested compounds.
For the calculation of the MG_MID, insensitive cell lines were
included with the highest concentration tested (Table 2).
been found as two broad singlets and doublet at
d ꢀ
9.42–9.49, d ꢀ 9.19–9.26, and d ꢀ 9.03–9.04 correspondingly.
This could be explained by amino-imino tautomerism of
these derivatives (3a–3d) [27].
Evaluation of anticancer activity in vitro
Synthesized 1-[2-(3,5-diaryl-4,5-dihydropyrazol-1-yl)-2-oxoethyl]-
1H-indole-2,3-diones 1a, 1b, 1d, and 1,3-dihydroindol-2-one
conjugates with 3,5-diaryl-4,5-dihydropyrazolyne and 4-thia-
zolidinone moieties (2d–2f, 3a, 3c, 3d) were submitted and
evaluated at the single concentration of 10ꢁ5 M towards
panel of approximately sixty cancer cell lines. The human
tumor cell lines were derived from nine different cancer
types: Leukemia, melanoma, lung, colon, CNS, ovarian, renal,
prostate, and breast cancers. Primary anticancer assays
(Table 1) were performed according to the US NCI protocol,
Table 1. Anticancer screening data in concentration 10.00 mM.
Compd.
60 cell lines assay in 1 dose 10.00 mM conc.
Mean growth % Range of growth % The most sensitive cell lines
Growth % of the most sensitive cell lines
1a
98.37
48.40–161.24
SR (Leukemia)
CCRF-CEM (Leukemia)
SR (Leukemia)
CCRF-CEM (Leukemia)
HL-60 (TB) (Leukemia)
K-562 (Leukemia)
MOLT-4 (Leukemia)
RPMI-8226 (Leukemia)
SR (Leukemia)
NCI-H522 (Non-small cell lung cancer)
HCT-116 (Colon cancer)
KM 12 (Colon cancer)
SW-620 (Colon cancer)
OVCAR-3 (Ovarian cancer)
PC-3 (Prostate cancer)
MCF-7 (Breast cancer)
MDA-MB-231/ATCC (Breast cancer)
PC-3 (Prostate cancer)
SNB-75 (CNS cancer)
SNB-75 (CNS cancer)
SR (Leukemia)
48.40
55.55
74.05
7.75
5.26
17.03
3.71
20.92
10.49
5.18
1b
1d
104.51
57.27
74.05–171.03
ꢁ31.51–121.19
5.58
13.70
ꢁ31.51
7.45
16.98
15.31
68.21
63.86
74.24
56.94
84.50
83.02
2d
2e
2f
3a
3c
3d
109.41
95.41
101.92
91.84
104.65
106.30
68.21–148.16
63.86–116.92
74.24–134.29
56.94–126.98
84.50–150.27
83.02–133.93
K-562 (Leukemia)
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Isatin-based Pyrazolines and Thiazolidines Conjugates
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Table 2. The influence of compound 1d on the growth of individual tumor cell lines (GI50 ꢂ 10.00 mM).
Disease
Cell line
GI50 (mM)
TGI (mM)
MG_MID
Leukemia
Leukemia
Leukemia
Leukemia
Leukemia
Leukemia
NSC lung cancer
NSC lung cancer
NSC lung cancer
NSC lung cancer
Colon cancer
Colon cancer
Colon cancer
Colon cancer
Colon cancer
CNS cancer
CNS cancer
CNS cancer
CNS cancer
Melanoma
Melanoma
Ovarian cancer
Ovarian cancer
Ovarian cancer
Ovarian cancer
Renal cancer
Renal cancer
Renal cancer
Renal cancer
Renal cancer
Prostate cancer
Breast cancer
Breast cancer
Breast cancer
28.39
0.75
2.64
3.35
2.52
0.69
1.69
10.0
3.69
8.08
2.59
2.00
3.40
3.74
2.98
2.19
8.90
3.32
3.05
3.57
2.70
8.15
4.61
2.05
2.28
4.27
2.82
5.68
2.25
4.96
7.77
2.26
2.61
8.46
2.32
61.52
11.7
11.9
13.5
8.21
8.53
5.71
27.3
17.3
>100.0
10.1
8.73
>100.0
>100.0
>100.0
5.92
>100.0
26.5
17.5
35.5
18.8
>100.0
>100.0
7.31
96.9
>100.0
18.2
>100.0
7.16
>100.0
>100.0
>100.0
12.2
CCRF-CEM
HL-60 (TB)
K-562
MOLT-4
RPMI-8226
SR
HOP-62
HOP-92
NCI-H23
NCI-H522
HCT-116
HCT-15
HT29
KM12
SW-620
SF-268
SF-539
SNB-75
U251
LOX IMVI
SK-MEL-2
IGROV1
OVCAR-3
OVCAR-4
OVCAR-8
786-0
CAKI-1
RXF 393
TK-10
UO-31
PC-3
MCF7
HS 578T
T-47D
48.6
>100.0
The tested compound 1d showed inhibition activity against
34 (GI50 < 10 mM) from 59 human tumor cells with average
GI50/TGI values 28.39 mM/61.52 mM (Table 2). With regard to
the sensitivity against some individual cell lines among several
subpanel, compound 1d showed the high cytostatic and cyto-
toxic effects against NSC lung cancer NCI-H522 cell line
(GI50 ¼ 2.59 mM, TGI ¼ 10.1 mM), colon cancer HCT-116
pound’s selectivity. Ratios between
3
and
6
refer to
moderate selectivity, ratios greater than 6 indicate high
selectivity toward the corresponding cell line, while com-
pounds not meeting either of these criteria are rated non-
selective [32]. In this context, the active compound 1d in the
present study was found to be high selectivity toward the
leukemia subpanel at both the GI50 and TGI levels (selectivity
index 14.63 and 6.19, respectively) and moderate selectivity
toward the CNS cancer and renal cancer subpanels with
selectivity index near 3 at the GI50 level (3.69 and 3.24,
respectively) (Table 3).
(GI50 ¼ 2.00 mM, TGI ¼ 8.73 mM) and SW-620 (GI50
¼
2.19 mM, TGI ¼ 5.92 mM) cell lines, ovarian cancer OVCAR-3
cell line (GI50 ¼ 2.05 mM, TGI ¼ 7.31 mM) and renal cancer
RXF 393 cell line (GI50 ¼ 2.25 mM, TGI ¼ 7.16 mM). It also
revealed an obvious sensitivity profile of 1d toward the all
leukemia subpanel tumor cell lines with GI50 values range
of 0.69–3.35 mM and the highest activity against CCRF-CEM
(GI50 ¼ 0.75 mM) and RPMI-8226 (GI50 ¼ 0.69 mM) cell lines.
The selectivity index (SI) obtained by dividing the full panel
MG-MID (mM) of the compound 1d by their individual sub-
panel MG-MID (mM) was considered as a measure of com-
The SAR study revealed that: (1) Hybridization of the isatin
ring system with the pyrazoline ring system can lead to
appearance of antitumor activity; (2) introduction of bromo
group in 5-position of isatin fragment enhanced the potency
of isatin-pyrazoline conjugates; (3) condensation of highly-
active isatin-pyrazoline hybrid (1d) with 4-thiazolidinones led
to complete loss of activity.
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Arch. Pharm. Chem. Life Sci. 2011, 344, 514–522
Table 3. Anticancer selectivity pattern of the most active compound
1d at the GI50 (mM) and TGI (mM) levels.
Conclusions
In the present paper new isatin based conjugates with pyrazo-
line and 4-thiazolidinone moieties were described. Antitumor
activity testing allowed us to identify highly active isatin-
pyrazoline hybrid 1d, as prospective anticancer agent with
high selective influence on the leukemia subpanel at both the
GI50 and TGI levels (selectivity index 14.63 and 6.19, respect-
ively) and moderate selectivity toward the CNS cancer and
renal cancer subpanels with selectivity index near 3 at the GI50
level. The highest activity compound 1d demonstrated on the
leukemia subpanel tumor cell lines with GI50 values range of
0.69–3.35 mM. The further investigations of such isatin deriva-
tives could be interesting with the hope to get more selective
anticancer agents among isatin-pyrazoline hydrid analoges.
Disease
GI50
SIa
TGI
9.9
72.7
84.7
53.6
81.8
78.7
72.5
SIb
Leukemia
1.9
45.4
61.6
7.7
56.5
17.2
8.8
14.63
0.68
0.46
3.69
0.50
1.65
3.24
0.89
1.16
6.19
0.85
0.73
1.15
0.75
0.78
0.84
0.62
0.85
NSC lung cancer
Colon cancer
CNS cancer
Melanoma
Ovarian cancer
Renal cancer
Prostate cancer
Breast cancer
31.8
24.5
>100.0
72.3
a
b
Selectivity index at the GI50 level. Selectivity index at the
TGI level.
Experimental
COMPARE analysis
NCI’s COMPARE algorithm (see e.g. http://dtp.nci.nih.gov)
[33, 34] allows to assume biochemical mechanisms of action
of novel compounds on the basis of their in-vitro activity profiles
when comparing with those of standard agents. We performed
COMPARE computations for synthesized highly-active com-
pound 1d against the NCI ‘‘Standard Agents’’ database at
the GI50 and TGI levels (Table 4). However obtained Pearson
correlation coefficients (PCC) preclude that the compound may
have the same mechanism of action as that of known anti-
cancer agents. The compound 1d showed the moderate corre-
lations at the TGI level with microtubulin formation-
modulating agents (microtubule polymerization inhibitors) –
macrolides rhizoxin and maytansine (Table 4). The isatin-pyr-
azoline conjugates do not belong to a class of these antitumor
antibiotics, however, according to the literature data, the
tubulin polymerization inhibitors were identified among isa-
tin [2, 35–37] and pyrazoline [13] derivatives. This fact prompts
us to further study of the antitumor activity mechanism of
isatin-pyrazoline conjugates.
Materials and methods
The starting 2-chloro-1-(3,5-diaryl-4,5-dihydropyrazol-1-yl)-etha-
nones [23], 2,4-thiazolidinedione [38], 2-amino-4-thiazolone
[38], 2-thioxo-4-thiazolidinone [39] were obtained according to
the methods described previously.
Melting points were measured in open capillary tubes on a
¨
BUCHI B-545 melting point apparatus and are uncorrected. The
elemental analyses (C, H, N) were performed using the Perkin-
Elmer 2400 CHN analyzer. Analyses indicated by the symbols of
the elements or functions were within ꢃ0.4% of the theoretical
values. The 1H-NMR spectra were recorded on Varian Gemini
300 MHz and 13C-NMR spectra on Varian Mercury-400 100 MHz
in DMSO-d6 or DMSO-d6 þ CCl4 mixture using tetramethylsilane
(TMS) as an internal standard. Chemical shifts are reported in
ppm units with use of d scale.
Chemistry
Synthesis of 1-[2-(3,5-diaryl-4,5-dihydropyrazol-1-yl)-2-
oxoethyl]-1H-indole-2,3-diones (1a–1d)
A mixture of 50 mmol isatin or 5-bromoisatin, 60 mmol of
appropriate 2-chloro-1-(3,5-diaryl-4,5-dihydropyrazol-1-yl)-etha-
Table 4. COMPARE analysis results for highly-active compound 1d.
End point PCCa Target
Target vector Target mechanism of actionb
NSC
GI50
TGI
0.509 Rifamycin
0.581 Rhizoxin
0.561 Maytansine
S133100
S332598
S153858
DNA-polymerase inhibitor
Microtubulin formation-modulating agent (microtubule polymerization inhibitor)
Microtubulin formation-modulating agent (microtubule polymerization inhibitor)
Reversible binding inhibitor of the human kinesin Eg5, antimitotic agent
Chloroalkylating agent
Chloroethylating alkylator, alkyl transferase-dependent cross-linkers
Heat shock protein (Hsp-90) inhibitor
0.533 L-Cysteine analogue S303861
0.529 CCNU
0.524 Methyl-CCNU
0.505 Macbecin II
S79037
S95441
S269148
a
b
Only correlations with PCC ꢄ 0.5 were selected, as significant. Putative mechanisms of action were identified with the use of
literature sources.
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Arch. Pharm. Chem. Life Sci. 2011, 344, 514–522
Isatin-based Pyrazolines and Thiazolidines Conjugates
519
none, and 100 mmol of potassium carbonate was stirred in
30 mL of DMF at room temperature during 12 h. The crystalline
products were separated by filtration, washed with water, etha-
nol, and dried. Recrystallization from mixture DMF/ethanol ren-
dered desired products in pure form.
d6 þ CCl4): d 7.81–7.77 (m, 3H, arom); 7.72 (s, 1H, arom); 7.37
(d, 1H, J ¼ 8.0 Hz, arom); 7.28 (d, 2H, J ¼ 8.0 Hz, arom); 7.11
(d, 1H, J ¼ 8.3 Hz, arom); 7.05 (d, 2H, J ¼ 8.4 Hz, arom); 5.58
(dd, 1H, J ¼ 11.8, 4.0 Hz, CH); 5.08 (d, 1H, J ¼ 17.5 Hz, CH2);
4.96 (d, 1H, J ¼ 17.5 Hz, CH2); 3.93 (dd, 1H, J ¼ 18.1,
11.8 Hz, CH2CH); 3.84 (s, 3H, CH3); 3.22 (dd, 1H, J ¼ 18.1,
13
4.0 Hz, CH2CH). C-NMR (100 MHz, DMSO-d ): d 163.5 (C O),
–
–
162.8 (C O), 158.8 (C O), 152.1 (C N), 142.2, 136.6, 135.2,
6
–
–
–
–
–
–
1-{2-[5-(4-Methoxyphenyl)-3-phenyl-4,5-dihydropyrazol-
133.8, 129.4, 129.0, 128.1, 123.8, 123.5, 114.7, 111.7, 59.9
(CHCH2), 55.8 (OCH3), 42.5 (CHCH2), 42.3 (CH2). Calcd.
for C26H19BrClN3O4: C, 56.49; H, 3.46; N, 7.60; Found: C, 56.58;
H, 3.38; N, 7.75%.
1-yl]-2-oxoethyl}-1H-indole-2,3-dione (1a)
Yield 68%, mp 236–2388C. IR [cmꢁ1]: 1736 (CO), 1664 (CO), 1616
(CO), 1448 (arom), 756. 1H-NMR (300 MHz, DMSO-d6 þ CCl4):
d 7.87–7.83 (m, 2H, arom); 7.65–7.48 (m, 5H, arom); 7.16–7.09
(m, 4H, arom); 6.87 (d, 2H, J ¼ 8.7 Hz, arom); 5.55 (dd, 1H,
J ¼ 11.6, 4.6 Hz, CH); 5.11 (d, 1H, J ¼ 17.5 Hz, CH2); 4.96 (d,
1H, J ¼ 17.5 Hz, CH2); 3.89 (dd, 1H, J ¼ 18.3, 11.6 Hz, CH2CH);
3.69 (s, 3H, CH3); 3.15 (dd, 1H, J ¼ 18.3, 4.6 Hz, CH2CH). 13C-NMR
General procedure for synthesis of 5-(1-{2-[3,5-diaryl-4,5-
dihydropyrazol-1-yl]-2-oxoethyl}-2-oxo-1,2-
dihydroindol-3-ylidene)-4-thiazolidinones (2) and 2-
–
–
–
–
(100 MHz, DMSO-d ): d 163.9 (C O), 161.8 (C O), 158.46 (C O),
–
–
6
–
151.8 (C N), 141.0, 138.8, 131.3, 129.2, 129.0, 128.0, 127.4, 124.8,
–
amino-4-thiazolones (3)
123.6, 114.6, 110.2, 60.2 (CHCH2), 55.5 (OCH3), 42.8 (CHCH2), 42.3
(CH2). Calcd. for C26H21N3O4: C, 71.06; H, 4.82; N, 9.56; Found:
C, 71.20; H, 4.70; N, 9.75%.
A mixtures of compound 1a–1d (3.3 mmol), appropriate 4-thia-
zolidinone derivative (3 mmol), and anhydrous sodium acetate
(3 mmol) were refluxed for 2 h in glacial acetic acid (10 mL).
Obtained powders were filtered off, washed with methanol, and
recrystallized with DMF/ethanol (1:2) mixture.
1-{2-[5-(4-Chlorophenyl)-3-phenyl-4,5-dihydropyrazol-
1-yl]-2-oxoethyl}-1H-indole-2,3-dione (1b)
Yield 75%, mp 256–2588C. IR [cmꢁ1]: 1742 (CO), 1658 (CO), 1632
(CO), 1456 (arom), 748. 1H-NMR (300 MHz, DMSO-d6 þ CCl4):
d 7.86–7.84 (m, 2H, arom); 7.62 (t, 1H, J ¼ 7.8 Hz, arom); 7.58
(d, 1H, J ¼ 7.4 Hz, arom); 7.54–7.50 (m, 3H, arom); 7.37 (d, 2H,
J ¼ 8.4 Hz, arom); 7.29 (d, 2H, J ¼ 8.4 Hz, arom); 7.15 (t, 1H,
J ¼ 7.4 Hz, arom); 7.11 (d, 1H, J ¼ 7.9 Hz, arom); 5.60 (dd, 1H,
J ¼ 11.8, 4.8 Hz, CH); 5.05 (d, 1H, J ¼ 17.4 Hz, CH2); 4.94 (d, 1H,
J ¼ 17.4 Hz, CH2); 3.89 (dd, 1H, J ¼ 18.3, 11.8 Hz, CH2CH); 3.25
(dd, 1H, J ¼ 18.3, 4.8 Hz, CH2CH). 13C-NMR (100 MHz, DMSO-d6):
5-(1-{2-[5-(4-Methoxyphenyl)-3-phenyl)-4,5-
dihydropyrazol-1-yl]-2-oxoethyl}-2-oxo-1,2-dihydroindol-3-
ylidene)-2-thioxo-4-thiazolidinone (2a)
Yield 71%, mp 290–2928C. IR [cmꢁ1]: 1740 (CO), 1680 (CO), 1612
(CO), 1520 (C C), 1440 (arom), 1232, 1184, 1088 (C S), 760. 1H-
–
–
–
–
NMR (300 MHz, DMSO-d6 þ CCl4): d 8.84 (br.s, 1H, NH); 7.85
(d, 2H, J ¼ 6.9 Hz, arom); 7.50–7.48 (m, 3H, arom); 7.42 (t, 1H,
J ¼ 7.6 Hz, arom); 7.16–7.13 (m, 3H, arom); 7.04 (d, 1H, J ¼ 7.7 Hz,
arom); 6.88 (d, 2H, J ¼ 6.9 Hz, arom); 5.53 (dd, 1H, J ¼ 11.4,
4.6 Hz, CH); 5.14 (d, 1H, J ¼ 17.2 Hz, CH2); 5.02 (d, 1H,
J ¼ 17.2 Hz, CH2); 3.90 (dd, 1H, J ¼ 18.0, 11.4 Hz, CH2CH); 3.74
(s, 3H, CH3); 3.15 (dd, 1H, J ¼ 18.0, 4.6 Hz, CH2CH). 13C-NMR
–
–
–
–
–
d 164.3 (C O), 163.8 (C O), 158.6 (C O), 156.4 (C N), 140.9, 138.9,
–
–
–
138.8, 131.2, 129.2, 129.0, 128.1, 127.4, 124.9, 123.8, 117.6, 111.7,
60.1 (CHCH2), 42.6 (CHCH2), 42.3 (CH2). Calcd. for C25H18ClN3O3:
C, 67.64; H, 4.07; N, 9.47; Found: C, 67.52; H, 3.98; N, 9.55%.
–
–
–
–
(100 MHz, DMSO-d ): d 188.8 (C S), 168.9 (C O), 163.3 (C O),
–
–
6
–
–
–
161.8 (C O), 151.8 (C N), 141.0, 138.7, 132.3, 129.2, 129.0,
–
1-{2-[5-(4-Chlorophenyl)-3-(4-methoxyphenyl)-4,5-
128.0, 124.8, 123.6, 123.5, 114.7, 110.7, 60.1 (CHCH2), 55.7
(OCH3), 42.7 (CHCH2), 42.3 (CH2). Calcd. for C29H22N4O4S2:
C, 62.80; H, 4.00; N, 10.10; Found: C, 62.87; H, 3.38; N, 10.19%.
dihydropyrazol-1-yl]-2-oxoethyl}-1H-indole-2,3-dione (1c)
Yield 71%, mp 245–2468C. IR [cmꢁ1]: 1725 (CO), 1658 (CO), 1632
(CO), 1462 (arom), 752. 1H-NMR (300 MHz, DMSO-d6 þ CCl4):
d 7.81 (d, 2H, J ¼ 8.6 Hz, arom); 7.63 (t, 1H, J ¼ 7.4 Hz, arom);
7.58 (d, 1H, J ¼ 7.0 Hz, arom); 7.39 (d, 2H, J ¼ 8.2 Hz, arom); 7.27
(d, 2H, J ¼ 8.2 Hz, arom); 7.14 (d, 2H, J ¼ 6.0 Hz, arom); 7.06
(d, 2H, J ¼ 8.6 Hz, arom); 5.57 (dd, 1H, J ¼ 11.8, 3.2 Hz, CH);
5.05 (d, 1H, J ¼ 17.6 Hz, CH2); 4.92 (d, 1H, J ¼ 17.6 Hz, CH2);
3.89 (dd, 1H, J ¼ 18.0, 11.8 Hz, CH2CH); 3.83 (s, 3H, CH3); 3.22
(dd, 1H, J ¼ 18.0, 3.2 Hz, CH2CH). 13C-NMR (100 MHz, DMSO-d6):
5-(1-{2-[5-(4-Chlorophenyl)-3-phenyl)-4,
5-dihydropyrazol-1-yl]-2-oxoethyl}-2-oxo-1,
2-dihydroindol-3-ylidene)-2-thioxo-4-thiazolidinone (2b)
Yield 67%, mp 270–2728C. IR [cmꢁ1]: 1740 (CO), 1680 (CO), 1608
(CO), 1536 (C C), 1440 (arom), 1232, 1180, 1088 (C S), 752. 1H-
–
–
–
–
NMR (300 MHz, DMSO-d6 þ CCl4): d 8.85 (br.s, 1H, NH); 7.82 (br.s,
2H, arom); 7.63 (t, 1H, J ¼ 7.6 Hz, arom); 7.50 (br.s, 3H, arom); 7.43
(t, 1H, J ¼ 7.7 Hz, arom); 7.37 (d, 2H, J ¼ 8.4 Hz, arom); 7.29 (d, 2H,
J ¼ 8.4 Hz, arom); 7.15 (t, 1H, J ¼ 7.3 Hz, arom); 7.09 (d, 1H,
J ¼ 7.7 Hz, arom); 5.61 (dd, 1H, J ¼ 11.4, 4.8 Hz, CH); 5.17 (d,
1H, J ¼ 17.3 Hz, CH2); 5.09 (d, 1H, J ¼ 17.3 Hz, CH2); 3.95 (dd,
–
–
–
–
–
d 163.5 (C O), 161.8 (C O), 158.6 (C O), 151.6 (C N), 141.1, 132.4,
–
–
–
129.4, 128.9, 128.0, 124.8, 123.8, 123.5, 114.7, 111.7, 59.9 (CHCH2),
55.8 (OCH3), 42.5 (CHCH2), 42.3 (CH2). Calcd. for C26H20ClN3O4:
C, 65.90; H, 4.25; N, 8.87; Found: C, 65.79; H, 4.18; N, 8.95%.
1H, J ¼ 17.8, 11.4 Hz, CH2CH); 3.15 (dd, 1H, J ¼ 17.8,
5-Bromo-1-{2-[5-(4-chlorophenyl)-3-(4-methoxyphenyl)-
4,5-dihydropyrazol-1-yl]-2-oxoethyl}-1H-indole-2,3-
dione (1d)
13
4.8 Hz, CH2CH). C-NMR (100 MHz, DMSO-d ): d 188.6 (C S),
–
–
168.8 (C O), 163.8 (C O), 160.8 (C O), 154.6 (C N), 140.8, 138.9,
6
–
–
–
–
–
–
–
–
138.8, 131.3, 129.2, 129.0, 128.2, 127.8, 124.8, 123.5, 117.7, 111.5,
60.1 (CHCH2), 42.6 (CHCH2), 42.3 (CH2). Calcd. for C28H19ClN4O3S2:
C, 60.16; H, 3.43; N, 10.02; Found: C, 60.29; H, 3.35; N, 10.14%.
Yield 78%, mp 258–2608C. IR [cmꢁ1]: 1740 (CO), 1668 (CO), 1608
(CO), 1444 (arom), 1256, 1176, 832. 1H-NMR (300 MHz, DMSO-
ß 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

520
D. Havrylyuk et al.
Arch. Pharm. Chem. Life Sci. 2011, 344, 514–522
1H-NMR (300 MHz, DMSO-d6 þ CCl4): d 8.99 (br.s, 1H, NH); 7.77–
7.71 (m, 3H, arom); 7.58 (br.s, 1H, arom); 7.35 (d, 2H, J ¼ 7.6 Hz,
arom); 7.26 (d, 2H, J ¼ 7.6 Hz, arom); 7.09–7.04 (m, 3H, arom);
5.57 (dd, 1H, J ¼ 10.8, 4.7 Hz, CH); 5.15 (d, 1H, J ¼ 17.8 Hz, CH2);
5.03 (d, 1H, J ¼ 17.8 Hz, CH2); 3.92 (dd, 1H, J ¼ 16.8,
5-(1-{2-[5-(4-Chlorophenyl)-3-(4-methoxyphenyl)-4,5-
dihydropyrazol-1-yl]-2-oxoethyl}-2-oxo-1,2-dihydroindol-3-
ylidene)-2-thioxo-4-thiazolidinone (2c)
Yield 75%, mp 288–2898C. IR [cmꢁ1]: 1745 (CO), 1682 (CO), 1615
(CO), 1542 (C C), 1445 (arom), 1180, 1088 (C S), 752. 1H-NMR
–
–
–
–
10.8 Hz, CH2CH); 3.84 (s, 3H, CH3); 3.20 (dd, 1H, J ¼ 16.8,
13
4.7 Hz, CH2CH). C-NMR (100 MHz, DMSO-d ): d 164.1 (C O),
(300 MHz, DMSO-d6 þ CCl4): d 8.82 (br.s, 1H, NH); 7.79 (d, 2H,
J ¼ 6.1 Hz, arom); 7.45 (t, 1H, J ¼ 7.2 Hz, arom); 7.37 (d, 2H,
J ¼ 7.8 Hz, arom); 7.25 (d, 2H, J ¼ 7.8 Hz, arom); 7.15 (t, 1H,
J ¼ 7.3 Hz, arom); 7.07–7.03 (m, 3H, arom); 5.57 (dd, 1H,
J ¼ 11.1, 4.2 Hz, CH); 5.15 (d, 1H, J ¼ 17.3 Hz, CH2); 5.03 (d, 1H,
J ¼ 17.3 Hz, CH2); 3.92 (dd, 1H, J ¼ 17.8, 11.1 Hz, CH2CH); 3.83 (s,
3H, CH3); 3.17 (dd, 1H, J ¼ 17.8, 4.2 Hz, CH2CH). 13C-NMR
–
–
162.6 (C O), 159.5 (C O), 157.2 (C O), 152.3 (C N), 142.4,
6
–
–
–
–
–
–
–
–
136.4, 135.4, 132.4, 129.2, 129.1, 128.1, 128.0, 123.7, 114.7,
59.9 (CHCH2), 55.8 (OCH3), 42.3 (CHCH2), 42.2 (CH2). Calcd.
for C29H20BrClN4O5S: C, 53.43; H, 3.09; N, 8.59; Found: C,
53.32; H, 3.15; N, 8.47%.
–
–
–
–
(100 MHz, DMSO-d ): d 188.8 (C S), 168.6 (C O), 163.7 (C O),
–
–
6
5-(1-{2-[5-(4-Methoxyphenyl)-3-phenyl)-4,5-
dihydropyrazol-1-yl]-2-oxoethyl}-2-oxo-1,2-dihydroindol-
–
–
161.0 (C O), 152.6 (C N), 141.2, 138.9, 132.4, 131.3, 129.4, 129.0,
–
–
128.0, 127.9, 124.8, 123.7, 114.8, 111.7, 60.0 (CHCH2), 55.8 (OCH3),
42.6 (CHCH2), 42.3 (CH2). Calcd. for C29H21ClN4O4S2: C, 59.13; H,
3.59; N, 9.51; Found: C, 59.02; H, 3.45; N, 9.34%.
3-ylidene)-2-amino-4-thiazolone (3a)
Yield 72%, mp 276–2788C. IR [cmꢁ1]: 2860, 1700 (CO), 1676 (CO),
1612 (CO), 1538 (C C), 1440 (arom), 1360, 1180. 1H-NMR
–
–
(300 MHz, DMSO-d6 þ CCl4): d 9.42, 9.21, 9.03 (br.s, s, d, 2H,
NH2); 7.85 (d, 2H, J ¼ 7.7 Hz, arom); 7.50–7.48 (m, 3H, arom);
7.35 (t, 1H, J ¼ 7.6 Hz, arom); 7.16 (d, 2H, J ¼ 8.5 Hz, arom); 7.12
(t, 1H, J ¼ 7.6 Hz, arom); 6.99 (d, 1H, J ¼ 7.7 Hz, arom); 6.88
(d, 2H, J ¼ 8.5 Hz, arom); 5.53 (dd, 1H, J ¼ 11.4, 4.3 Hz, CH);
5.15 (d, 1H, J ¼ 17.3 Hz, CH2); 5.03 (d, 1H, J ¼ 17.3 Hz, CH2);
3.93 (dd, 1H, J ¼ 18.1, 11.4 Hz, CH2CH); 3.74 (s, 3H, CH3); 3.22
(dd, 1H, J ¼ 18.1, 4.3 Hz, CH2CH). 13C-NMR (100 MHz, DMSO-d6): d
5-(5-Bromo-1-{2-[5-(4-chlorophenyl)-3-(4-methoxy-
phenyl)-4,5-dihydropyrazol-1-yl]-2-oxoethyl}-2-oxo-1,2-
dihydroindol-3-ylidene)-2-thioxo-4-thiazolidinone (2d)
Yield 70%, mp 284–2868C. IR [cmꢁ1]: 1740 (CO), 1672 (CO), 1608
–
–
–
(CO), 1520 (C C), 1432 (arom), 1232, 1176, 1088 (C S), 832.
–
1H-NMR (300 MHz, DMSO-d6 þ CCl4): d 9.01 (br.s, 1H, NH);
7.81–7.77 (m, 3H, arom); 7.60 (s, 1H, arom); 7.37 (d, 2H,
J ¼ 7.6 Hz, arom); 7.25 (d, 2H, J ¼ 7.6 Hz, arom); 7.06–7.02
(m, 3H, arom); 5.54 (dd, 1H, J ¼ 11.2, 3.4 Hz, CH); 5.15 (d, 1H,
J ¼ 17.1 Hz, CH2); 5.02 (d, 1H, J ¼ 17.1 Hz, CH2); 3.87 (dd, 1H,
–
–
–
–
–
163.9 (C O), 158.9 (C O), 156.2 (C O), 152.4 (C N), 143.8, 134.1,
–
–
–
131.5, 131.2, 131.0, 129.2, 128.0, 127.3, 122.7, 120.2, 114.5, 109.6,
60.2 (CHCH2), 55.5 (OCH3), 42.9 (CHCH2), 42.3 (CH2). Calcd.
for C29H23N5O4S: C, 64.79; H, 4.31; N, 13.03; Found: C, 64.88;
H, 4.39; N, 13.19%.
J ¼ 17.8, 11.2 Hz, CH2CH); 3.83 (s, 3H, CH3); 3.19 (dd, 1H, J ¼ 17.8,
13
3.4 Hz, CH2CH). C-NMR (100 MHz, DMSO-d ): d 188.6 (C S),
–
–
168.2 (C O), 163.5 (C O), 162.0 (C O), 152.2 (C N), 142.0,
6
–
–
–
–
–
–
–
–
136.9, 135.2, 133.7, 131.4, 129.2, 128.2, 127.9, 124.7, 123.8,
114.8, 111.8, 59.9 (CHCH2), 55.7 (OCH3), 42.5 (CHCH2), 42.3
(CH2). Calcd. for C29H20BrClN4O4S2: C, 52.14; H, 3.02; N, 8.39;
Found: C, 52.22; H, 2.95; N, 8.31%.
5-(1-{2-[5-(4-Chlorophenyl)-3-phenyl)-4,5-
dihydropyrazol-1-yl]-2-oxoethyl}-2-oxo-1,2-dihydroindol-
3-ylidene)-2-amino-4-thiazolone (3b)
Yield 69%, mp 280–2828C. IR [cmꢁ1]: 2850, 1688 (CO), 1670 (CO),
1608 (CO), 1536 (C C), 1440 (arom), 1362, 1180. 1H-NMR
–
–
5-(1-{2-[5-(4-Chlorophenyl)-3-(4-methoxyphenyl)-4,5-
dihydropyrazol-1-yl]-2-oxoethyl}-2-oxo-1,2-dihydroindol-3-
ylidene)-2,4-thiazolidinedione (2e)
(300 MHz, DMSO-d6 þ CCl4): d 9.19, 9.04 (br.s, d, 2H, NH2); 7.83
(d, 2H, J ¼ 7.7 Hz, arom); 7.59–7.47 (m, 3H, arom); 7.34 (br.s, 3H,
arom); 7.27 (d, 2H, J ¼ 7.7 Hz, arom); 7.17 (t, 1H, J ¼ 7.5 Hz,
arom); 6.98 (d, 1H, J ¼ 7.2 Hz, arom); 5.61 (dd, 1H, J ¼ 11.3,
4.5 Hz, CH); 5.15 (d, 1H, J ¼ 17.2 Hz, CH2); 5.03 (d, 1H,
J ¼ 17.2 Hz, CH2); 3.93 (dd, 1H, J ¼ 18.1, 11.3 Hz, CH2CH); 3.22
(dd, 1H, J ¼ 18.1, 4.3 Hz, CH2CH). 13C-NMR (100 MHz, DMSO-d6):
Yield 69%, mp 248–2508C. IR [cmꢁ1]: 1736 (CO), 1705 (CO), 1672 (CO),
1
1612 (CO), 1536 (C C), 1448 (arom), 1256, 1176, 832, 752. H-NMR
–
–
(300 MHz, DMSO-d6 þ CCl4): d 8.83 (br.s, 1H, NH); 7.79 (d, 2H,
J ¼ 8.4 Hz, arom); 7.63 (t, 1H, J ¼ 7.4 Hz, arom); 7.58 (d, 1H,
J ¼ 7.8 Hz, arom); 7.38 (d, 2H, J ¼ 7.6 Hz, arom); 7.27 (d, 2H,
J ¼ 7.6 Hz, arom); 7.15–7.09 (m, 2H, arom); 7.05 (d, 2H,
J ¼ 8.4 Hz, arom); 5.58 (dd, 1H, J ¼ 11.1, 3.2 Hz, CH); 5.08 (d, 1H,
J ¼ 17.6 Hz, CH2); 4.95 (d, 1H, J ¼ 17.6 Hz, CH2); 3.93 (dd, 1H,
–
–
–
–
–
d 163.7 (C O), 159.2 (C O), 158.8 (C O), 151.8 (C N), 143.0, 138.9,
–
–
–
138.0, 131.7, 131.2, 131.0, 129.4, 128.0, 127.3, 124.7, 123.2, 117.5,
109.6, 60.0 (CHCH2), 42.7 (CHCH2), 42.3 (CH2). Calcd.
for C28H20ClN5O3S: C, 62.05; H, 3.72; N, 12.92; Found: C, 62.19;
H, 3.58; N, 12.74%.
J ¼ 17.7, 11.1 Hz, CH2CH); 3.84 (s, 3H, CH3); 3.17 (dd, 1H, J ¼ 17.7,
13
3.2 Hz, CH2CH). C-NMR (100 MHz, DMSO-d ): d 163.5 (C O), 161.7
–
–
(C O), 158.7 (C O), 156.2 (C O), 151.5 (C N), 141.0, 138.8, 132.3,
6
–
–
–
–
–
–
–
–
5-(1-{2-[5-(4-Chlorophenyl)-3-(4-methoxyphenyl)-4,5-
dihydropyrazol-1-yl]-2-oxoethyl}-2-oxo-1,2-dihydroindol-
3-ylidene)-2-amino-4-thiazolone (3c)
129.1, 129.0, 128.0, 124.8, 123.8, 123.5, 114.7, 111.7, 59.9 (CHCH2),
55.8 (OCH3), 42.5 (CHCH2), 42.3 (CH2). Calcd. for C29H21ClN4O5S: C,
60.79; H, 3.69; N, 9.78; Found: C, 60.87; H, 3.55; N, 9.64%.
Yield 79%, mp 269–2718C. IR [cmꢁ1]: 2860, 1694 (CO), 1672 (CO),
1608 (CO), 1538 (C C), 1434 (arom), 1356, 1180. 1H-NMR
–
–
5-(5-Bromo-1-{2-[5-(4-chlorophenyl)-3-
(4-methoxyphenyl)-4,5-dihydropyrazol-1-yl]-2-oxoethyl}-2-
(300 MHz, DMSO-d6 þ CCl4): d 9.49, 9.26, 9.03 (br.s, s, d, 2H,
NH2); 7.96 (d, 2H, J ¼ 8.4 Hz, arom); 7.65 (t, 1H, J ¼ 7.4 Hz, arom);
7.59 (d, 1H, J ¼ 7.4 Hz, arom); 7.38 (d, 2H, J ¼ 7.6 Hz, arom); 7.25
(d, 2H, J ¼ 7.6 Hz, arom); 7.12–7.08 (m, 2H, arom); 7.03 (d, 2H,
J ¼ 8.4 Hz, arom); 5.56 (dd, 1H, J ¼ 11.2, 4.3 Hz, CH); 5.14 (d, 1H,
oxo-1,2-dihydroindol-3-ylidene)-2,4-thiazolidinedione (2f)
Yield 75%, mp 280–2828C. IR [cmꢁ1]: 1744 (CO), 1704 (CO), 1676
–
(CO), 1608 (CO), 1520 (C C), 1460 (arom), 1312, 1256, 1180, 824.
–
ß 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

Arch. Pharm. Chem. Life Sci. 2011, 344, 514–522
Isatin-based Pyrazolines and Thiazolidines Conjugates
521
J ¼ 17.3 Hz, CH2); 5.02 (d, 1H, J ¼ 17.3 Hz, CH2); 3.90 (dd, 1H,
13
which is the drug concentration resulting in a 50% lower net
protein increase in the treated cells (measured by SRB stain-
ing) as compared to the net protein increase seen in the
control cells. The drug concentration resulting in total
growth inhibition (TGI) was calculated from Ti ¼ Tz. The
LC50 (concentration of drug resulting in a 50% reduction in
the measured protein at the end of the drug treatment as
compared to that at the beginning) indicating a net loss of
cells following treatment was calculated from:
J ¼ 18.2, 11.2 Hz, CH2CH); 3.83 (s, 3H, CH3); 3.14 (dd, 1H, J ¼ 18.2,
–
4.3 Hz, CH2CH). C-NMR (100 MHz, DMSO-d ): d 163.8 (C O),
–
–
159.9 (C O), 158.6 (C O), 151.6 (C N), 143.2, 141.0, 138.2,
6
–
–
–
–
–
132.7, 131.2, 131.0, 129.4, 128.0, 124.8, 123.8, 123.5, 115.0,
110.9, 60.0 (CHCH2), 55.8 (OCH3), 42.6 (CHCH2), 42.3 (CH2).
Calcd. for C29H22ClN5O4S: C, 60.89; H, 3.88; N, 12.24; Found:
C, 61.02; H, 3.75; N, 12.36%.
5-(5-Bromo-1-{2-[5-(4-chlorophenyl)-3-(4-methoxy-
phenyl)-4,5-dihydropyrazol-1-yl]-2-oxoethyl}-2-oxo-1,2-
dihydroindol-3-ylidene)-2-amino-4-thiazolone (3d)
½ðTi ꢁ TzÞ=Tzꢅ ꢆ 100 ¼ ꢁ50
Yield 70%, mp 275–2768C. IR [cmꢁ1]: 2856, 1696 (CO), 1672 (CO),
Values were calculated for each of these three parameters if the
level of activity is reached; however, if the effect was not reached
or was exceeded, the value for that parameter was expressed as
more or less than the maximum or minimum concentration
tested. The lowest values are obtained with the most sensitive cell
lines. Compounds having GI50 ꢂ 100 mM were declared to be
active.
1608 (CO), 1536 (C C), 1432 (arom), 1360, 1180. 1H-NMR
–
–
(300 MHz, DMSO-d6 þ CCl4): d 9.44, 9.23 (br.s, s, 2H, NH2); 7.97
(s, 1H, arom); 7.76 (d, 2H, J ¼ 8.4 Hz, arom); 7.52 (d, 1H,
J ¼ 8.3 Hz, arom); 7.35 (d, 2H, J ¼ 8.2 Hz, arom); 7.25 (d, 2H,
J ¼ 8.2 Hz, arom); 7.04 (d, 2H, J ¼ 8.4 Hz, arom); 6.98 (d, 1H,
J ¼ 8.3 Hz, arom); 5.57 (dd, 1H, J ¼ 11.4, 4.6 Hz, CH); 5.14
(d, 1H, J ¼ 17.2 Hz, CH2); 5.01 (d, 1H, J ¼ 17.2 Hz, CH2); 3.91
(dd, 1H, J ¼ 18.1, 11.4 Hz, CH2CH); 3.84 (s, 3H, CH3); 3.22 (dd,
1H, J ¼ 18.1, 4.6 Hz, CH2CH). 13C-NMR (100 MHz, DMSO-d6):
We are grateful to Dr. V.L. Narayanan from Drug Synthesis and Chemistry
Branch, National Cancer Institute, Bethesda, MD, USA, for in vitro
evaluation of anticancer activity.
–
–
–
–
–
d 164.1 (C O), 159.2 (C O), 158.8 (C O), 151.8 (C N), 142.2,
–
–
–
136.8, 135.2, 132.7, 131.2, 129.4, 128.1, 127.8, 123.8, 114.8,
60.0 (CHCH2), 55.8 (OCH3), 42.7 (CHCH2), 42.3 (CH2). Calcd. for
C29H21BrClN5O4S: C, 53.51; H, 3.25; N, 10.76; Found: C, 53.38;
H, 3.37; N, 10.69%.
The authors have declared no conflict of interest.
Pharmacology
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