Synthesis and evaluation of indole, pyrazole, chromone and pyrimidine based conjugates for tumor growth inhibitory activities - Development of highly efficacious cytotoxic agents

Article abstract of DOI:10.1016/j.ejmech.2010.08.004

Based upon the lead compounds 10 and 11, a number of conjugates were synthesized by the combination of chromone-pyrimidine, chromone-indolinone, chromone-pyrazole, indole-pyrimidine, indole-indolinone and indole-pyrazole moieties. Evaluation of these compounds for tumor growth inhibitory activities over 60 human tumor cell lines provided highly efficacious compounds 15, 41, 43, 66, 69, and 72 with an average GI₅₀ over all the 60 human tumor cell lines as 3.2 μM, 3.1 μM, 1.7 μM, 2.6 μM, 50.1 μM and 2.0 μM, respectively.

Full text of DOI:10.1016/j.ejmech.2010.08.004

European Journal of Medicinal Chemistry 45 (2010) 4968e4982
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Synthesis and evaluation of indole, pyrazole, chromone and pyrimidine based
conjugates for tumor growth inhibitory activities e Development of highly
efficacious cytotoxic agents
,
a
b
Palwinder Singh a , Matinder Kaur , Wolfgang Holzer
*
a Department of Chemistry, Guru Nanak Dev University, Amritsar-143005, India
b Institute of Pharmaceutical Chemistry, University of Vienna, Althanstrasse-14, A-1090 Wein, Austria
a r t i c l e i n f o
a b s t r a c t
Article history:
Based upon the lead compounds 10 and 11, a number of conjugates were synthesized by the combination
of chromone-pyrimidine, chromone-indolinone, chromone-pyrazole, indole-pyrimidine, indole-indoli-
none and indole-pyrazole moieties. Evaluation of these compounds for tumor growth inhibitory activ-
ities over 60 human tumor cell lines provided highly efficacious compounds 15, 41, 43, 66, 69, and 72
Received 17 May 2010
Received in revised form
28 July 2010
Accepted 5 August 2010
Available online 12 August 2010
with an average GI50 over all the 60 human tumor cell lines as 3.2
and 2.0 M, respectively.
mM, 3.1 mM, 1.7 mM, 2.6 mM, 50.1 mM
m
Ó 2010 Elsevier Masson SAS. All rights reserved.
Keywords:
Cancer
Hybrid molecules
Tumor growth inhibition
1. Introduction
inhibitor) [24], mitoxantrone (5, GI50 ¼ 0.059
[25e27], indirubin (6 IC50 ¼ 10 M for CDK1) [28,29], celecoxib (7,
GI50 ¼ 15.9 M, COX-2 inhibitor) [30e33], and hydrazinecarbo-
thioamide (8, GI50 ¼ 9.17 M, DNA antimetabolite) [34] (Chart 1) have
mM, topoII inhibitor)
m
As the number of deaths due to cancer alone is more than those
caused by AIDS, malaria, and tuberculosis combined, it is turning into
one of the most overwhelming health problems worldwide. In the
last 30 years [1,2] the number of patients with tumors get doubled, by
2020 it will double once more, and by 2030 it will be triple and if new
treatments are not found, by 2030, there will be 27 million people
with cancer and 17 million deaths annually. The practice of chemo-
therapy of cancer suffers from various drawbacks viz. the participa-
tion of a number of enzymes like ribonucleotide reductase (RNR),
thymidylate synthase (TS), thymidylate phosphorylase (TP), top-
oisomerase II (topoII) etc. at different stages of development of cancer
[3,4], survival of cancer cells even under anaerobic conditions [5e7],
and ultimately the problem of multidrug resistance [8e14] devel-
oped in the cancerous cells towards chemotherapeutic agents. The
understanding about the participation of the enzymes at various
stages of cancer led to the development of a number of anticancer
agents, some of which are currently in clinical use. The drugs like
m
m
created some hope for the life of cancer patients. Flavonoids, [35] the
most common group of polyphenolic compounds in the human diet
and found ubiquitously in plants, are also known for significant
anticancer activity [36]. Therefore, the available anticancer drugs
have distinct mechanism of action which may vary in their effects on
different types of normal and cancer cells. However, the chemo-
therapeutic agents under present use suffer from various drawbacks,
such as their toxicity to normal cells, bone marrow depression, the
adverse effect to the gastrointestinal tract [37] and above all the cost
factor. This undoubtedly underscores the need of developing new
chemotherapeutic agents for more effective and economical treat-
ment of cancer.
Compounds 10 and 11 [38] (Chart 2) were identified as highly
promising candidates for tumor growth inhibitory activities out of
which compound 11 showed significantly high maximum tolerable
dose (MTD) and presently under investigation for in-vivo anticancer
activities. Owing to the importance of hybrid molecules [39e41] and
to improve the anticancer activities of compounds 10 and 11, new
compounds have been developed (Chart 2) by combining the
appropriate fragments of some of the anticancer drugs shown in
Chart 1. Some pyrazoles and indolinones-carrying pyridine, naphthyl
and styryl moieties were also included in the present investigations.
5-fluorouracil (1, GI50 ¼ 17.1
m
M, TS inhibitor) [15,16], indomethacin
M,
M, topoII
(2, GI50 ¼ 64.3
m
M, COX-2 inhibitor) [17e20], taxol (3, GI50 ¼ 0.01
m
topoII inhibitor) [21e23], doxorubicin (4, GI50 ¼ 0.096
m
* Corresponding author. Fax: þ91 183 2258819.
E-mail address: palwinder_singh_2000@yahoo.com (P. Singh).
0223-5234/$ e see front matter Ó 2010 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejmech.2010.08.004

P. Singh et al. / European Journal of Medicinal Chemistry 45 (2010) 4968e4982
4969
O
O
OH
O
O
H3CO
O
OH
OH
O
O
O
N
F
O
NH
O
OH
HN
O
O
O
O
N
H
Cl
5-Fluorouracil, 1
Indomethacin, 2
Taxol, 3
H
O
O
OH
O
OH
O
N
OH
OH
OH
OH
O
O
NH
NH
O
H
HN
N
H
O
OH
OH
OH
N
O
H
NH2
Mitoxantrone, 5
Indirubin, 6
Doxorubicin, 4
F3C
CH3
N
N
N
CH=NNHCSNH2
SO2NH2
Celecoxib, 7
HO
Hydrazinecarbothioamide, 8
Chart 1. Some clinically used anticancer drugs.
Tumor growth inhibitory activities of these compounds were tested
at National Cancer Institute (NCI), Bethesda, USA and led to the
identification of new molecules which showed better in-vitro
activities than 5-fluorouracil, indomethacin and celecoxib.
dose levels. The fibers were collected from the mice on the day
following the fourth compound treatment and subjected to the
stable endpoint MTT (3-[4,5-dimethylthiazole-2-yl]-2,5-diphe-
nyltetrazolium bromide) assay. The optical density of each sample
was determined spectrophotometrically at 540 nm and the mean
of each treatment group was calculated. Out of the maximum
possible score of 96 for an agent (12 cell lines ꢀ 2 sites ꢀ 2 dose
levels ꢀ 2 [score]), compound 10 showed IP score 2 (out of 48), SC
score 6 (out of 48) and it did not cause cell killing. In-vivo investi-
gation on compound 11 is underway.
2. Results and discussion
2.1. MTD test on compounds 10 and 11
The MTD of compounds 10 and 11 (Chart 2) was determined by
Nontumored Animal Toxicity Assay. Intraperitoneous (IP) injections
of 400 mg/kg, 200 mg/kg and 100 mg/kg were given to the female
athymic nude mice and monitored their weight loss. It was
observed that after 19 days there was no survival where compound
10 was injected indicating that all the three doses of compound 10
were toxic while in case of compound 11, there was no loss and
therefore its MTD is 400 mg/kg. The experiment was repeated with
compound 10, this time two doses of 50 mg/kg and 25 mg/kg were
given through IP injection to two mice. Now there was no loss after
17 days indicating 50 mg/kg as the MTD of compound 10 (Table S1).
2.3. Synthesis and tumor growth inhibitory activities of new
conjugates
Encouraged by the tumor growth inhibitory activities and
toxicity data of compounds 10 and 11, a library of compounds was
synthesized by the combination of two biologically active hetero-
cycles. Since microwave heating [42,43] is continuing to be an
attractive application and has become widely accepted non-
conventional energy source for performingorganic reactions due to
the dramatically short reaction time, improved yields and envi-
ronmentally benign reactions; here also, the CeC bond formation
between the two selected moieties were carried under micro-
waves. In the synthesis of indole-based compounds, Knoevenegal
condensation under microwave irradiation was preceded by the
N-substitution of indole.
2.2. Hollow fiber assay
The preliminary in-vivo activity of compound 10 was demon-
strated by hollow fiber assay, which provided quantitative indices
of drug efficacy of this compound. A panel of 12 tumor cell lines viz.
NCI-H23, NCI-H522, MDA-MB-231, MDA-MB-435, SW-620, COLO
205, LOX, UACC-62, OVCAR-3, OVCAR-5, U251 and SF-295 was used
(supplementary information, Table S2). Based on the MTD, each
mice was administered by IP and subcutaneous (SC) injection at 2
For the first set of compounds, the pyrimidine/indolinone/pyr-
azole moiety was combined with chromone. A finely ground
mixture of 3-formyl chromone (12, 1 mmol) and barbituric acid
(1.2 mmol) on irradiating in microwave oven for 1 min gave product

4970
P. Singh et al. / European Journal of Medicinal Chemistry 45 (2010) 4968e4982
R
O
N
O
O
O
O
N
R
R
O
O
N
N
Cl
N
O
O
O
O
N
R
Cl
10, R=H
11, R=CH3
Chromone-pyrimidine
Chromone-indolinone
O
O
CH3
N
N
O
N
O
N
R
R
Cl
Indole-indolinone
CH3
Chromone-pyrazole
CH3
R
N
N
R
O
N
Cl
N
O
N
N
R
O
Cl
Cl
Cl
N
OH
Indole-pyrazole
R=
Chart 2. New designed Hybrid molecules.
13 in 80% yield as creamish white solid, mp >250 ꢁC. Similar
reactions of 12 with 1,3-dimethylbarbituric acid, indolinone and
pyrazole provided the required products 14, 15 and 16 respectively
(Scheme 1).
Compounds 14, 15 and 16 were investigated for their tumor
growth inhibitory activities at 10ꢂ5 M concentration on all the 60
human tumor cell lines. Compounds 14 and 16 showed 30e40%
inhibition only at RPMI-8226, SR, M14 and UO-31 cell lines of
O
O
CH3
O
O
O
H
N
O
N
O
N
O
CH3
O
O
M
H
O
N
N
N
O
O
N
e
O
CH3
H
O
O
N
H
14
13
CH3
e
t
u
n
i
W
80%, >250 0C
83%, 185 0C
m
I
1
,
,
I
1
W
m
O
O
M
i
n
u
t
CHO
CH3
N
O
N
12
Cl
N
N
a
O
e
H
C
O
t
u
Cl
n
i
m
3
,
Cl
1
,
M
I
W
W
O
I
M
CH3
O
Cl
N
O
O
Cl
N
N
O
O
Cl
15
78%, >250 0C
16
87%, 227 0C
Scheme 1. Synthesis of chromone-based compounds.

Table 1
GI50 M) and LC50 (mM) values for compounds 15, 41, 43, 66, 69 and 72.
(m
Panel/cell line
15
41
43
66
69
72
GI50
LC50
GI50
LC50
GI50
LC50
GI50
LC50
GI50
LC50
GI50
LC50
Leukemia
CCRF-CEM
HL-60 (TB)
K-562
MOLT-4
RPMI-8226
SR
2.0
2.0
2.0
2.9
1.2
1.4
>100
>100
>100
>100
69.1
>100
0.34
>100
>100
>100
>100
>100
>100
2.3
1.5
2.2
1.5
1.9
2.3
>100
3.1
3.1
2.8
3.1
2.8
2.3
>100
>100
>100
>100
>100
>100
97.7
>100
44.6
>100
>100
e
>100
>100
>100
>100
>100
e
3.2
3.8
3.9
4.4
2.2
e
>100
>100
>100
>100
>100
e
2.4
3.1
2.8
3.4
2.2
9.3
>100
>100
>100
>100
Non-small cell lung cancer
A549/ATCC
EKVX
4.2
2.8
4.7
>100
>100
>100
e
3.2
2.8
12.8
1.5
7.4
4.1
2.9
1.9
4.2
>100
>100
>100
>100
83.1
50.1
>100
2.6
1.6
1.3
1.9
1.6
1.6
1.9
1.6
1.8
>100
>100
16.2
3.0
3.1
4.6
2.6
2.3
28.1
1.6
2.6
>100
>100
57.5
>100
64.5
>100
>100
6.4
>100
>100
>100
31.6
>100
>100
>100
e
1.9
2.0
2.3
0.01
e
43.6
>100
89.1
9.1
HOP-62
5.4
6.3
7.7
6.9
54.9
6.9
HOP-92
e
e
NCI-H226
NCI-H23
NCI-H322M
NCI-H460
NCI-H522
20.8
4.4
5.8
2.0
e
>100
>100
>100
>100
e
47.8
>100
>100
>100
>100
e
e
>100
>100
>100
e
3.5
2.8
1.8
1.6
>100
28.8
7.9
54.9
7.0
7.5
70.7
Colon cancer
COLO 205
HCC-2998
HCT-116
HCT-15
HT29
KM12
SW-620
3.8
4.2
1.9
0.6
3.0
3.0
5.2
>100
>100
>100
>100
>100
>100
>100
7.6
6.0
1.6
3.4
3.8
1.8
2.9
54.9
52.4
6.9
>100
54.9
8.9
1.8
1.6
1.2
1.1
1.8
1.8
1.9
5.8
5.7
5.0
7.4
6.3
7.4
6.6
2.3
1.9
1.5
2.2
2.9
1.6
1.7
>100
>100
e
>100
e
2.3
e
>100
e
6.3
6.1
>100
>100
5.8
6.7
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
1.7
2.0
2.1
1.8
1.9
6.9
28.8
9.1
6.3
6.7
52.4
CNS cancer
SF-268
SF-295
SF-539
SNB-19
SNB-75
U251
3.3
4.2
3.0
e
>100
>100
>100
e
2.4
2.2
5.0
7.2
e
57.5
66.1
66.1
1.5
1.7
1.5
2.7
1.6
1.5
8.1
7.0
5.7
63.1
36.3
8.5
2.3
3.1
2.5
5.6
7.0
1.7
>100
97.7
22.9
0.01
45.70
0.9
>100
>100
>100
>100
>100
>100
3.6
1.7
2.4
2.2
1.4
1.8
40.7
13.8
>100
32.3
32.3
7.4
>100
>100
>100
>100
>100
1.9
1.4
>100
>100
e
3.4
54.9
23.9
40.7
Melanoma
LOX IMVI
MALME-3M
M14
MDA-MB-435
SK-MEL-2
SK-MEL-28
SK-MEL-5
UACC-257
UACC-62
1.5
2.4
5.2
2.6
2.7
3.8
2.4
4.5
2.1
57.5
1.6
2.6
2.3
2.2
e
6.6
58.8
33.8
51.2
1.5
1.0
e
6.0
7.2
e
1.4
2.7
1.6
2.5
3.8
2.0
2.2
2.1
1.7
6.7
>100
7.5
>100
>100
8.9
20.8
>100
>100
29.5
>100
>100
21.8
>100
>100
>100
>100
>100
e
1.7
1.9
1.7
1.8
2.3
e
6.4
16.9
7.7
61.6
24.5
>100
>100
>100
1.7
1.9
2.1
1.5
2.3
1.3
6.9
6.7
5.4
5.4
9.1
5.2
61.6
e
e
>100
>100
>100
>100
e
e
e
e
7.6
5.0
4.9
45.7
95.4
61.6
>100
>100
39.8
>100
>100
>100
1.6
1.7
1.6
5.7
18.2
6.9
8.5
Ovarian cancer
IGROV1
OVCAR-3
OVCAR-4
OVCAR-5
OVCAR-8
NCI/ADR-RES
SK-OV-3
1.6
2.6
3.6
20.8
5.4
3.3
97.9
e
e
e
1.2
1.8
2.3
e
8.5
7.7
38.9
1.6
2.1
3.3
2.8
2.6
3.3
3.2
16.5
8.9
>100
>100
>100
>100
>100
26.9
58.8
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
1.7
1.7
3.7
2.4
2.1
3.0
3.4
>100
>100
>100
>100
>100
>100
1.6
e
8.1
6.9
>100
>100
>100
>100
8.9
4.1
3.4
8.3
>100
>100
>100
>100
e
e
3.2
e
70.7
35.4
2.1
>100
26.3
74.1
Renal cancer
786e0
A498
3.0
4.3
4.1
2.4
1.8
3.3
3.3
1.8
>100
>100
>100
>100
3.3
1.2
3.1
1.9
4.1
3.9
10.4
1.1
91.2
21.8
>100
>100
43.6
70.7
>100
48.9
1.7
3.1
2.3
1.4
2.0
1.5
2.1
1.3
5.8
7.5
39.8
>100
7.5
5.4
27.5
7.4
2.1
17.7
1.7
1.9
2.1
2.4
3.1
1.6
10.2
>100
6.1
>100
7.2
21.3
>100
16.9
>100
>100
>100
>100
>100
e
1.9
2.9
1.8
1.3
e
6.6
72.4
7.2
ACHN
CAKI-1
RXF 393
SN12C
TK-10
UO-31
7.4
69.1
e
e
>100
>100
>100
>100
>100
>100
34.6
>100
>100
>100
>100
2.2
3.0
1.4
58.8
50.1
6.0
7.5
Prostate cancer
PC-3
DU-145
1.3
4.6
>100
>100
2.4
2.7
>100
46.7
1.3
1.7
6.4
6.1
2.9
2.8
>100
53.7
>100
70.7
>100
>100
2.2
2.1
93.3
9.1
Breast cancer
MCF7
MDA-MB- 31/ATCC
HS 578T
BT-549
T-47D
3.2
3.8
9.1
2.8
3.0
2.4
3.2
>100
>100
>100
>100
>100
>100
e
e
1.7
1.5
e
8.1
6.4
1.1
2.2
1.6
1.6
2.2
1.6
2.6
>100
>100
>100
6.1
>100
39.8
45.7
e
e
e
e
3.1
2.2
2.4
4.1
4.4
3.1
>100
>100
37.1
>100
69.1
58.8
36.3
32.3
27.5
>100
>100
>100
>100
>100
>100
2.0
5.6
2.4
2.3
0.9
2.0
29.5
e
e
>100
23.4
>100
7.4
27.5
1.6
1.9
e
11.2
28.8
>100
17.7
50.1
MDA-MB-468
Mean
97.7
1.7
14.1

4972
P. Singh et al. / European Journal of Medicinal Chemistry 45 (2010) 4968e4982
CH3
N
N
O
CH3
N
CHO
CHO
NaH, ACN
RX
Cl
N
N
R
O
N
R
N
H
MWI
Cl
26-34
17-25
Scheme 2. Synthesis of indole-pyrazole conjugates.
Leukemia, melanoma and renal cancer (Table S3). Compound 15
showed 50e90% tumor growth inhibition in most of the cell lines
with the mean of percent growth over all the cell lines as 40.79
(Table S3). It was further tested at 10ꢂ4 M, 10ꢂ5 M, 10ꢂ6 M, 10ꢂ7 M
and 10ꢂ8 M concentrations and found to exhibit an average GI50
(Scheme 5, Table 4) and the combination of indole-3-carboxaldehye
with oxindole [48,49] followed by the introduction of various
substituents at nitrogen of two heterocycles resulted in the forma-
tion of compounds 52e60 (Scheme 6, Table 5).
Compounds 38, 40, 41, 43, 44, 46, 49, 54, 55, 56, 58 and 59 were
investigated by NCI for their tumor growth inhibitory activities.
Compounds 38 and 40 showed 30e60% inhibition with their mean
of growth percent as 88.85 and 97.46 respectively (Table S4).
Compound 41 with 2,6-dichlorobenzoyl group showed notable
results with 33e82% tumor growth inhibition and specificity for
certain cell lines like CCRF-CEM, HOP-92, HCT-116, KM12, LOX IMVI,
(over all the 60 cell lines) 3.2
specificity for certain cell lines like RPMI-8226, SR, HCT-15, U251,
LOX IMVI and PC-3 where it exhibited GI50 1.2 M, 1.4 M, 0.6 M,
1.4 M,1.5 M, and 1.3 M, respectively (Table 1). LC50 of compound
15 was 97.7 M indicating very low toxicity of this compound.
mM. Compound 15 also showed
m
m
m
m
m
m
m
Therefore, on the basis of appreciable tumor growth inhibitory
activity of compound 15, it seems that the presence of indole
moiety as one of the two heterocycles of these hybrid molecules is
essential. In the next sets of molecules, following compounds 10, 11
and 15, the indole moiety was retained and combined with pyr-
azole/indolinone heterocycles.
Treatment of indole-3-carboxaldehyde with NaH in CH3CN fol-
lowed by the addition of alkyl/acyl/tosyl halide provided N-
substituted indole-3-caboxaldehydes 18e25 (Scheme 2). Further
reaction of compounds 17e25 (1 mmol) with pyrazole (1.2 mmol)
in microwave oven gave solid compounds 26e34 in 1e5 min
(Scheme 2, Table 2).
Compounds 26, 27, 29, 30, 32 and 33 were investigated at
10ꢂ5 M concentration for tumor growth inhibitory activities over 60
human tumor cell lines. Compounds 26, 27, 29, 32 and 33 did not
exhibit anticancer activities as the mean of percent growth of
tumor cells over all the 60 cell lines is almost 100 in presence of
these compounds (Table S3). Though the mean of percent growth of
tumor cells in presence of compound 30 is 89.25, it showed 83%
inhibition of tumor growth at NCI-H226 cell line of Non-small cell
lung cancer (Table S3). Therefore, the results of the six compounds
investigated in this set indicated that the presence of benzyl group
at N ꢂ 1 of indole (compound 30) is leading to some activity in the
compound. To see if the presence of more benzyl groups improves
the activity of the compound, 10 was modified to compound 35 by
introducing benzyl groups at N ꢂ 1 and N ꢂ 3 of pyrimidine moiety
(Scheme 3). Compound 35 showed 43e75% tumor growth inhibi-
tion at NCI-H460, COLO 205, SF-295, SNB-75, UACC-257, UACC-62,
OVCAR-4, SK-OV-3, RXF 393, TK-10, T-47D cell lines of Non-small
cell lung cancer, colon, central nervous system (CNS), melanoma,
ovarian, renal, and breast cancer with a mean of growth percent of
74.71 (Table S3). Less tumor growth inhibitory activity of
compound 35 than that of compounds 10 and 11 indicates that the
presence of many hydrophobic substituents in the compound may
not be desirable for anticancer activities.
OVCAR-3, A489, CAKI-1 and UO-31 where it exhibited GI50 0.34
1.5 M, 1.6 M, 1.8 M, 1.6 M, 1.6 M, 1.2 M, 1.9 M, and 1.1
m
m
M,
M,
m
m
m
m
m
m
m
respectively (Table 1). Compound 43 showed 75e99% tumor
growth inhibition in most of the cell lines with mean of growth
percent inhibition as ꢂ18.77% and average GI50 (over all the 60 cell
lines) 1.7 mM. Compound 43 also showed specificity for HL-60 (TB),
MOLT-4, HOP-62, HCT-116, SF-268, SF-539, U251, LOX IMVI,
MALME-3M, SK-MEL-3M, UACC-62, IGROV1, CAKI-1, UO-31, and PC-
3 cell lines with GI50 in the range 1.2 mMe1.5 mM (Table 1). The
tumor growth inhibition by compounds 44, 46 and 49 range from
20 to 66% at HOP-92, HCT-15, OVCAR-4, A498, UO-31, PC-3 and
T-47D cell lines of Non-small cell lung cancer, colon, ovarian, renal,
prostate and breast cancer with their mean of growth percent
inhibitions as 84.76, 88.62, and 102.10, respectively (Table S4).
Compounds 54, 55, 56, 58 and 59 exhibited 33e76% tumor growth
inhibition at certain cell lines with a mean of growth percent as
99.11, 88.44, 88.46, 75.24, and 94.54, respectively. Therefore, after
the combination of two indole moieties, it is the nature of the
substituent which also contributes to the activity of the
compounds. Here, the presence of 2,6-dichlorobenzoyl and 2,6-
dichlorophenyl substituent at one of the two nitrogens led to better
tumor growth inhibitory activities of compounds 41 and 43 in
comparison to their analogues.
Two more categories of compounds were prepared by intro-
ducing appropriate substituents on pyrazole and indolinone
moieties. A mixture of pyrazole and an aldehyde after irradiation
under microwaves provided the desired products (66e70) in
quantitative yields (Scheme 7, Table 6). The mixture of indolinone
and aldehydes (as used in Scheme 7) on irradiating under micro-
waves in presence of NaHCO3 provided compounds 71e74 (Scheme
8, Table 7). It is noteworthy that the Knoevenegal condensation of
indolinone (formation of 15, Schemes 5 and 8) requires NaHCO3.
Compounds 66, 67, 69 and 72 were investigated for their tumor
growth inhibitory activities. Compound 67 did not produce desir-
able results, the mean of percent growth of tumor cells over all the
60 cell lines was 108.97 (Table S5). Compounds 66, 69 and 72
showed 74e99%, 50e99% and 87e99% inhibition in most of the cell
lines with the mean of percent growth over all the cell lines as
33.13, 25.40, and 5.01, respectively (Table S5). These were passed to
The third category of compounds was built by the combination of
two indole moieties (indirubin and sunitinib analogues [44e47]).
Reactions of N-substituted indole-3-carboxaldehydes and oxindole
gave compounds 36e42 (Scheme 4, Table 3), N-substituted indole-
3-carboxaldehyde and indolinone provided compounds 43e51

P. Singh et al. / European Journal of Medicinal Chemistry 45 (2010) 4968e4982
Table 2 (continued)
4973
Table 2
Percentage yields and melting points for indoleepyrazole conjugates 26e34.
%
Mp
Reactant
R
Product
Reactant
R
Product
%
Mp
yield (ꢁC)
yield (ꢁC)
CH3
N
CH3
N
N
N
O
25
88
105
N
N
H
O
17
H
70
89
>240
Cl
Cl
26
27
34
CH3
N
further testing at 10ꢂ4 M, 10ꢂ5 M, 10ꢂ6 M, 10ꢂ7 M and 10ꢂ8
concentrations and exhibited an average GI50 (over all the 60 cell
lines) 2.6 M, 50.1 M, and 2.0 M, respectively. Compound 66 also
M
N
N
O
18
145
Cl
m
m
m
showed specificity for certain cell lines like SR, NCI-H23, NCI-H460,
COLO 205, HCC-2998, HCT-116, HCT-15, KM12, SW-620, SF-268,
U251, LOX IMVI, M14, SK-MEL-28, SK-MEL-5, UACC-257, UACC-62,
IGROV1, OVCAR-3, 786-0, ACHN, CAKI-1, RXF 393, UO-31, MCF7,
MDA-MB-231/ATCC, HS 578T, BT-549, T-47D, and MDA-MB-468
CH3
N
N
N
O
19
86
193
with GI50 in the range 1.1
mMe2.3
mM (Table 1). Compound 69
Cl
28
showed GI50 0.01 M at SF-539 cell line of CNS cancer and 0.9
m
m
M at
SNB-75 cell line of CNS cancer. Compound 72 also showed speci-
ficity for RPMI-8226, A549/ATCC, EKVX, HOP-92, NCI-H460, NCI-
H522, COLO 205, HCT-116, HCT-15, HT29, KM12, SW-620, SF-295,
SNB-19, SNB-75, U251, LOX IMVI, MALME-3M, M14, MDA-MB-435,
SK-MEL-2, SK-MEL-5, UACC-257, UACC-62, IGROV1, OVCAR-3,
OVCAR-8, 786-0, ACHN, CAKI-1, SN12C, UO-31, PC-3, DU-145, MDA-
MB-231/ATCC, and MDA-MB-468 cell lines with GI50 ranging from
CH3
N
O
S
O
N
N
S
O
O
O
215
e218
20
86
Cl
CH3
0.01
mM to 2.3 mM (Table 1). A comparison of the anticancer
CH3
29
activities of compounds 66, 67, 69, and 72 showed that the
combination of cinnamyl-pyrazole and pyridinyl-indolinone, in the
form of compounds 66 and 72, result in better anticancer activities.
CH3
N
N
N
O
2.4. Docking studies
21
95
182
Cl
In order to support the experimental results, the docking [50] of
the most active compounds (15, 41, 43, 66 and 72) in the active sites
of RNR, TS, TP and COX-2 were performed (supporting information,
Figs. S1eS20). It was found that each compound shows interactions
with more than one enzyme out of RNR, TS, TP and COX-2.
30
CH3
N
N
N
O
O
3. Conclusions
22
87
>240
O
Cl
Investigations of a series of indole, pyrimidine, pyrazole and
chromone based conjugates for tumor growth inhibitory activities
have identified compounds 15, 41, 43, 66, and 72 with GI50 (over 60
31
CH3
N
human tumor cell lines) in the range 1.7 mMe3.2 mM and in-vitro
therapeutic indices (LC50/GI50) 30.5, 18.9, 10.0, 17.5, and 13.7,
respectively. It seems as indole/indolinone along with the presence
of benzyl/2,6-dichlorophenyl/2,6-dichlorobenzoyl substituent is
essential as one of the two heterocyclic moieties of these hybrid
molecules (because all of them except compounds 66 and 69 carry
indole/indolinone as one of the two fragments) for their appre-
ciable tumor growth inhibitory activities.
N
N
O
O
O
23
80
>240
Cl
Cl
Cl
32
CH3
N
4. Experimental
Cl
N
N
O
Cl
4.1. Chemistry
O
O
24
94
>240
Cl
Cl
Cl
Melting points were determined in capillaries and are uncor-
rected. 1H and 13C NMR spectra were recorded on JEOL 300 MHz
and 75 MHz NMR spectrometer, respectively using CDCl3 and/or
DMSO-d6 as solvent. Chemical shifts are given in ppm with TMS as
33

4974
P. Singh et al. / European Journal of Medicinal Chemistry 45 (2010) 4968e4982
O
O
N
NH
K2CO3, DMF
N
O
N
O
Benzyl bromide
stir, r.t
N
O
N
H
O
35
10
Scheme 3. Synthesis of compound 35.
an internal reference. J-values are given in Hertz. Signals are
abbreviated as singlet, s; doublet, d; double-doublet, dd; triplet, t;
multiplet, m. In 13C NMR spectral data, þve, ꢂve terms correspond
to CH3, CH, CH2 signals in DEPT-135 NMR spectra. Chromatography
was performed with silica 100e200 mesh and reactions were
monitored by thin layer chromatography (TLC) with silica plates
coated with silica gel HF-254. The mass spectra were recorded
using JEOL SX102/Da-600 instrument. Elemental analysis was
performed on Thermoelectron FLASH EA1112 CHN analyzer. Reac-
tions under microwaves were performed using domestic micro-
wave oven (INALSA model 1MW17EG) with microwave power
700 W and operating frequency 2450 MHz.
Preparation of N-substituted indole-3-carboxaldehyde (18e25)
(Procedure A). N-substituted indole-3-carboxaldehyde was prepared
by the treatment of indole-3-carboxaldehyde (1 mmol) with NaH
(1.5 mmol) in CH3CN followed by the addition of alkyl/acyl/tosyl
halides (1.5 mmol). After the completion of the reaction as indicated
via TLC, the reaction mixture was filtered and the filtrate was
evaporated to give solid compounds in quantitative yields.
Preparation of conjugate molecules 26e42, 66e70 (Procedure B).
A finely ground mixture of indole-3-carboxaldehyde (17e25)
(1 mmol) and 1-(3-chlorophenyl)-3-methyl-2-pyrazolin-5-one/
oxindole (1.2 mmol) on irradiating in microwave oven gave solid
products which were washed with diethyl ether to get pure prod-
ucts 26e42. Similar reactions of 1-(3-chlorophenyl)-3-methyl-2-
pyrazolin-5-one with cinnamaldehyde/pyridine-2 carboxaldehyde/
p-nitrobenzaldehyde/naphthaldehyde/anthranaldehyde (1 mmol)
gave products 66e70.
Table 3
Percentage yields and melting points for indoleeoxindole conjugates 36e42.
Reactant
R
Product
% yield Mp (ꢁC)
N
H
N
O
18
90
83
212
180
36
37
N
H
N
O
19
20
O
S
O
N
H
N
O
S
O
O
76
232
CH3
CH3
38
39
N
H
N
O
O
22
23
85
83
>240
O
Preparation of conjugates 43e51 and 71e74 (Procedure C). Indole-
3-carboxaldehyde (17e25)(1mmol)andindolinone(1.2mmol)were
irradiated under microwaves in the presence of catalytic amount of
NaHCO3. The reaction mixture was extracted with ethyl acetate/
chloroform (CHCl3). The combined organic layers were dried over
sodium sulphate and concentrated by evaporating the solvent to give
the desired products 43e51. Similar reactions of indolinone with
cinnamaldehyde/pyridine-2 carboxaldehyde/p-nitrobenzaldehyde/
anthranaldehyde (1 mmol) gave products 71e74.
N
H
N
O
O
>240
Cl
O
Cl
40
41
Preparation of indole-oxindole conjugates 52e60 (Procedure D). A
finely ground mixture of indole-3-carboxaldehyde (17e25)
Cl
N
H
N
O
Cl
O
24
25
80
89
196
230
O
Cl
Cl
CHO
N
O
H
N
H
N
N
R
O
N
O
N
R
MWI
H
18-20, 22-25
36-42
42
Scheme 4. Synthesis of indole-oxindole conjugates.

P. Singh et al. / European Journal of Medicinal Chemistry 45 (2010) 4968e4982
4975
Table 4
Percentage yields and melting points for indoleeindolinone conjugates 43e51.
N
Cl
, NaHCO3
MWI
O
Product % yield
Mp (ꢁC)
CHO
N
Cl
Cl
N
R
O
Cl
N
R
N
N
Cl
Cl
N
H
O
95
242
178
Cl
17-25
43-51
43
44
Scheme 5. Synthesis of indole-indolinone conjugates.
(1 mmol) and oxindole (1.2 mmol) on irradiating in microwave
oven gave solid product which was washed with diethyl ether to
get pure product 52. Further the N-substitutions of compound 52
were performed following the procedure A to get solid compounds
53e60.
N
O
83
85
Cl
N
4.1.1. 5-((4-Oxo-4H-chromen-3-yl)methylene)pyrimidine-2,4,6
(1H,3H,5H)-trione (13)
Cl
N
O
168
Cl
Creamish white solid, 80% yield, mp >250 ꢁC; nmax (KBr): 1713
(C]O), 3380 (NH); FAB mass m/z 285 (M þ 1); (Found: C 58.97, H
2.78, N 9.60; C14H8N2O5 requires C 59.16, H 2.84, N 9.86).
45
4.1.2. 1,3-Dimethyl-5-((4-oxo-4H-chromen-3-yl)methylene)
pyrimidine-2,4,6(1H,3H,5H)-trione (14)
N
N
S
Cl
O
O
O
Cl
Creamish solid, 83% yield, mp 185 ꢁC; nmax (KBr): 1650 (C]O);
78
>240
1H NMR (300 MHz, CDCl3):
d
3.45 (s, 6H, 2ꢀCH3), 7.46e7.55 (m, 2H,
ArH), 7.73 (dd, J3 ¼ 7.2 Hz, J4 ¼ 1.5 Hz, 1H, ArH), 8.28 (dd, J3 ¼ 7.8 Hz,
CH3
J4 ¼ 1.5 Hz, 1H, ArH), 8.88 (s, 1H), 9.79 (s, 1H, 2-H); 13C NMR
46
(normal/DEPT-135) (CDCl3):
d
28.4 (þve, CH3), 29.0 (þve, CH3),
117.9 (þve, CH), 118.34 (þve, CH), 118.36 (þve, CH), 123.5 (þve, CH),
126.3 (þve, CH), 126.5 (þve, CH), 134.5 (C), 148.7 (C), 151.0 (C), 155.6
(C), 161.2 (C), 161.7 (C), 163.50 (C), 175.2 (C); FAB mass m/z 312
(M þ 1); (Found: C 60.86, H 4.13, N 9.27; C16H12N2O5 requires C
61.54, H 3.87, N 8.97).
N
Cl
N
O
81
90
232
170
Cl
47
4.1.3. (3Z)-1-(2,6-Dichlorophenyl)-3-((4-oxo-4H-chromen-3-yl)
methylene)indolin-2-one (15)
N
Cl
N
O
Orange solid, 78% yield, mp >250 ꢁC; nmax (KBr): 1720, 1680 (C]
Cl
O); 1H NMR (300 MHz, CDCl3):
d
6.44 (d, J ¼ 7.8 Hz, 1H, ArH), 7.16 (t,
O
48
J ¼ 7.6 Hz,1H, ArH), 7.24 (t, J ¼ 7.7 Hz,1H, ArH), 7.41 (t, J ¼ 8.2 Hz,1H,
ArH), 7.46 (t, J ¼ 7.6 Hz,1H, ArH), 7.51 (d, J ¼ 7.8 Hz,1H, ArH), 7.54 (d,
J ¼ 8.2 Hz, 2H, ArH), 7.71 (t, J ¼ 7.6 Hz, 1H, ArH), 7.77 (d, J ¼ 7.6 Hz,
1H, ArH), 8.18 (s, 1H), 8.32 (d, J ¼ 8.1 Hz1H, ArH), 10.29 (s, 1H, ArH);
13C NMR (normal/DEPT-135) (CDCl3):
d
109.0 (þve, CH), 109.1 (C),
N
Cl
N
O
118.3 (þve, CH),118.5 (C),120.1 (C),122.9 (þve, CH),123.0 (þve, CH),
123.7 (þve, CH), 124.2 (þve, CH), 124.7 (C), 125.4 (þve, CH), 125.7
(þve, CH), 126.4 (þve, CH), 127.3 (þve, CH), 127.9 (þve, CH), 128.9
(þve, CH), 129.0 (C), 129.1 (C), 130.4 (þve, CH), 130.8 (þve, CH),
134.1 (þve, CH), 135.7 (C), 140.2 (C), 156.0 (C), 161.3 (C), 165.6 (C),
176.0 (C). FAB mass m/z 434 (M þ 1), 434:436:438 (9:6:1); (Found:
C 63.73, H 3.24, N 3.75; C24H13Cl2NO3 requires C 66.38, H 3.02,
N 3.23).
75
82
>260
Cl
O
Cl
49
N
Cl
N
O
Cl
250
Cl
O
Cl
50
4.1.4. (4Z)-1-(3-Chlorophenyl)-3-methyl-4-((4-oxo-4H-chromen-
3-yl)methylene)-1H-pyrazol-5(4H)-one (16)
Orange solid, 87% yield, mp 227 ꢁC; nmax (KBr): 1705 (C]O); 1H
N
Cl
N
O
NMR (300 MHz, CDCl3):
d 2.39 (s, 3H, CH3), 7.17e7.18 (m, 1H, ArH),
91
190
Cl
7.33 (t, J ¼ 8.1 Hz,1H, ArH), 7.50e7.58 (m, 2H, ArH), 7.74e7.79 (m,1H,
ArH), 7.88e7.91 (m, 1H, ArH), 8.01e8.04 (m, 2H, ArH), 8.29 (dd,
J3 ¼ 8.1 Hz, J4 ¼ 1.5 Hz, 1H, ArH), 10.75 (s, 1H); 13C NMR (normal/
51
DEPT-135) (CDCl3):
d
22.5 (þve, CH3), 117.93 (þve, CH), 118.33 (þve,
CH), 122.3 (þve, CH), 123.59 (þve, CH), 126.5 (þve, CH), 130.3
(þve, CH), 130.6 (þve, CH), 135.2 (þve, CH), 150.5 (þve, CH), 155.6

4976
P. Singh et al. / European Journal of Medicinal Chemistry 45 (2010) 4968e4982
N
CHO
O
H
RX
N
N
N
O
N
O
NaH,ACN
R
H
R
MWI
H
N
H
53-60
52
Scheme 6. Synthesis of indole-oxindole conjugates.
(þve, CH), 118.38 (C), 126.3 (C), 134.53 (C), 138.7 (C), 148.5 (C), 161.2
(C), 165.7 (C), 175.2 (C); FAB mass m/z 365 (M þ 1), 365:367 (3:1);
(Found: C 66.13, H 3.73, N 7.70; C20H13ClN2O3 requires C 65.85, H
3.59, N 7.68).
J3 ¼ 7.8 Hz, J4 ¼ 2.1 Hz,1H, ArH), 8.05 (dd, J3 ¼ 6.6 Hz, J4 ¼ 2.4 Hz,1H,
ArH), 8.10 (t, J ¼ 2.1 Hz, 1H, ArH), 9.90 (s, 1H, indole2-H); 13C NMR
(normal/DEPT-135) (CDCl3):
d
13.1 (þve, CH3), 21.6 (þve, CH3), 114.0
(C), 115.2 (þve, CH), 116.6 (þve, CH), 118.2 (þve, CH), 118.7 (þve,
CH), 124.3 (þve, CH), 124.5 (þve, CH), 125.2 (þve, CH), 125.6 (C),
127.3 (þve, CH),129.81 (þve, CH),129.85 (þve, CH),130.2 (þve, CH),
133.1 (þve, CH), 134.4 (C), 134.5 (C), 134.6 (C), 136.0 (C), 139.5 (C),
145.9 (þve, CH), 150.3 (C), 162.5 (C); FAB mass m/z 490 (M þ 1),
4.1.5. (4Z)-4-((1H-Indol-3-yl)methylene)-1-(3-chlorophenyl)-3-
methyl-1H-pyrazol-5(4H)-one (26)
Reddish orange solid, 70% yield, mp >240 ꢁC; nmax (KBr): 1640
(C]O), 3200 (NH); 1H NMR (300 MHz, CDCl3):
d
2.38 (s, 3H, CH3),
490:492 (3:1); (Found: C 63.59, H 4.00, N 8.40, S 6.47;
7.16e7.35 (m, 6H, ArH), 7.37 (t, J ¼ 8.7 Hz, 2H, ArH), 7.85e8.00 (m,
1H, ArH), 8.00 (t, J ¼ 2.1 Hz, 1H, indole2-H); FAB mass m/z 335 (Mþ),
335:337 (3:1); (Found: C 67.77, H 4.24, N 12.45; C19H14ClN3O
requires C 67.96, H 4.20, N 12.51).
C26H20ClN3O3S requires C 63.73, H 4.11, N 8.58, S 6.54).
4.1.9. (4Z)-4-((1-benzyl-1H-indol-3-yl)methylene)-1-(3-
chlorophenyl)-3-methyl-1H-pyrazol-5(4H)-one (30)
Reddish orange solid, 95% yield, mp 182 ꢁC; nmax (KBr): 1660
4.1.6. (4Z)-4-((1-Allyl-1H-indol-3-yl)methylene)-1-(3-
chlorophenyl)-3-methyl-1H-pyrazol-5(4H)-one (27)
(C]O); 1H NMR (300 MHz, CDCl3):
d 2.42 (s, 3H, CH3), 5.49 (s, 2H,
CH2), 7.13e7.35 (m, 9H, ArH), 7.84 (s, 1H, ¼H), 7.88e8.11 (m, 4H
Shiny orange solid, 89%yield, mp 145 ꢁC; nmax (KBr): 1680 (C]O);
ArH), 9.98 (s, 1H, indole2-H); 13C NMR (normal/DEPT-135)
1H NMR (300 MHz, CDCl3):
d
2.41 (s, 3H, CH3), 4.8 (dd, J2 ¼ 5.4 Hz,
(CDCl3):
d
13.1 (þve, CH3), 51.5 (ꢂve, CH2), 109.4 (C), 111.4 (þve,
J3 ¼1.8 Hz, 2H, CH2), 5.3 (ddd, J2 ¼16.8 Hz, J3 ¼10.2 Hz, J4 ¼ 0.3 Hz, 2H,
CH2), 6.05 (multiplet of 12 lines, 1H, CH), 7.11 (dd, J3 ¼ 6.9 Hz,
J4 ¼ 0.9 Hz, 1H, ArH), 7.14e7.45 (m, 4H, ArH), 7.82 (s, 1H, ¼H),
7.88e7.90 (m,1H, ArH), 7.96 (dd, J3 ¼ 6.6 Hz, J4 ¼ 0.9 Hz,1H, ArH), 8.11
(t, J ¼ 2.1 Hz, 1H, ArH), 9.85 (s, 1H, indole2-H); 13C NMR (normal/
CH), 112.1 (C), 116.6 (þve, CH), 118.0 (þve, CH), 118.7 (þve, CH),
119.5 (C), 120.7 (C), 122.7 (þve, CH), 123.8 (þve, CH), 124.1 (þve,
CH), 126.3 (C), 126.7 (þve, CH), 128.1 (þve, CH), 128.9 (þve, CH),
129.3 (C), 129.7 (þve, CH), 129.8 (þve, CH), 134.4 (C), 134.5 (C),
135.2 (C), 135.4 (þve, CH), 136.6 (C), 139.9 (C), 141.0 (þve, CH),
150.8 (C), 161.3 (C), 163.3 (C); FAB mass m/z 426 (M þ 1), 426:428
(3:1); (Found: C 72.94, H 4.88, N 9.58; C26H20ClN3O requires C
73.32, H 4.73, N 9.87).
DEPT-135) (CDCl3):
d
13.1 (þve, CH3), 50.1 (ꢂve, CH2),111.2 (C),112.0
(þve, CH), 116.7 (ꢂve, CH2), 118.0 (þve, CH), 118.8 (þve, CH), 118.9
(þve, CH), 119.3 (þve, CH), 122.7 (þve, CH), 123.7 (þve, CH), 124.1
(þve, CH), 129.2 (C), 129.7 (C), 131.4 (C), 134.4 (þve, CH), 135.5 (C),
136.6 (C),140.0 (þve, CH),140.7 (C),150.9 (C),163.4 (C); FAB mass m/z
375 (Mþ), 375:377 (3:1); (Found: C 69.97, H 4.74, N 10.95;
C22H18ClN3O requires C 70.30, H 4.83, N 11.18).
4.1.10. Compound 31
Reddish orange solid, 87% yield, mp >240 ꢁC; nmax (KBr): 1670
(C]O); 1H NMR (300 MHz, CDCl3):
d 2.43 (s, 3H, CH3), 7.11e7.14 (m,
1H, ArH), 7.22e7.52 (m, 4H, ArH), 7.61e7.74 (m, 5H, ArH), 7.86e8.01
(m, 3H, ArH), 8.46e8.49 (m, 1H, ¼H), 9.96 (s, 1H, indole2-H); FAB
mass m/z 441 (M þ 2), 441:443 (3:1); (Found: C 70.74, H 4.32, N 9.80;
C26H18ClN3O2 requires C 70.99, H 4.12, N 9.55).
4.1.7. (4Z)-1-(3-Chlorophenyl)-3-methyl-4-((1-(prop-2-ynyl)-1H-
indol-3-yl)methylene)-1H-pyrazol-5(4H)-one (28)
Reddish solid, 86% yield, mp 193 ꢁC; nmax (KBr): 1650 (C]O); 1H
NMR (300 MHz, CDCl3):
d
2.41 (s, 3H, CH3), 2.52 (t, J ¼ 2.7 Hz, 1H,
hH), 5.03 (d, J ¼ 2.7 Hz, 2H, CH2), 7.14 (dd, J3 ¼ 6.9, J4 ¼ 0.9 Hz, 1H,
ArH), 7.25e7.43 (m, 4H, ArH), 7.55 (dd, J3 ¼ 5.1 Hz, J4 ¼ 1.5 Hz, 1H,
ArH), 7.78 (s,1H, ¼H), 7.88 (dd, J3 ¼ 4.2 Hz, J4 ¼ 2.7 Hz,1H, ArH), 7.96
(dd, J3 ¼ 6.6 Hz, J4 ¼ 0.9 Hz, 1H, ArH), 9.91 (s, 1H, indole2-H); 13C
4.1.11. Compound 32
Reddish orange solid, 80% yield, mp >240 ꢁC; nmax (KBr): 1620
(C]O); 1H NMR (300 MHz, CDCl3):
d 2.43 (s, 3H, CH3), 7.13e7.16 (m,
1H, ArH), 7.25e7.52 (m, 4H, ArH), 7.60e8.01 (m, 7H, ArH),
8.43e8.47 (m, 1H, ¼H), 9.92 (s, 1H, indole2-H); FAB mass m/z 473
(Mþ), 473:475:477 (9:6:1); (Found: C 65.48, H 3.54, N 8.53;
C26H17Cl2N3O2 requires C 65.83, H 3.61, N 8.86).
NMR (normal/DEPT-135) (CDCl3):
d
13.1 (þve, CH3), 37.2 (ꢂve, CH2),
75.2 (þve, CH), 75.8 (C), 110.9 (C), 112.2 (þve, CH), 116.6 (þve, CH),
118.1 (þve, CH), 118.7 (þve, CH), 120.0 (þve, CH), 122.9 (þve, CH),
124.0 (þve, CH), 124.2 (C), 129.2 (þve, CH), 129.7 (þve, CH), 134.4
(C), 135.3 (þve, CH), 136.1 (C), 139.9 (C), 150.8 (C), 163.3 (C); FAB
mass m/z 373 (Mþ), 373:375 (3:1); (Found: C 70.49, H, 4.53, N 11.02;
C22H16ClN3O requires C 70.68, H 4.31, N, 11.24).
4.1.12. Compound 33
Reddish orange solid, 94% yield, mp >240 ꢁC; nmax (KBr): 1680
(C]O); 1H NMR (300 MHz, CDCl3):
d 2.36 (s, 3H, CH3), 7.06e7.50 (m,
8H, ArH), 7.70e7.78 (m, 3H,ArH), 8.57(s,1H, ¼H), 9.44(s,1H,indole2-
H); FAB mass m/z 508 (Mþ), 508:510:512:514 (27:27:9:1); (Found: C
61.58, H 3.14, N 8.33; C26H16Cl3N3O2 requires C 61.38, H 3.17, N 8.26).
4.1.8. (4Z)-1-(3-Chlorophenyl)-3-methyl-4-((1-tosyl-1H-indol-3-
yl)methylene)-1H-pyrazol-5(4H)-one (29)
Orange solid, 86% yield, mp 215e18 ꢁC; nmax (KBr): 1660 (C]O);
1H NMR (300 MHz, CDCl3):
d
2.35 (s, 3H, CH3), 2.43 (s, 3H, CH3), 7.16
4.1.13. (4Z)-1-(3-Chlorophenyl)-4-((1-cinnamyl-1H-indol-3-yl)
methylene)-3-methyl-1H-pyrazol-5(4H)-one (34)
(dd, J3 ¼ 7.8 Hz, J4 ¼ 0.9 Hz, 1H, ArH), 7.27 (d, J ¼ 8.4 Hz, 2H, ArH),
7.34e7.44 (m, 3H, ArH), 7.61 (s, 1H, ¼H), 7.75 (dd, J3 ¼ 6.45 Hz,
J4 ¼ 2.7 Hz, 1H, ArH), 7.94 (d, J ¼ 8.4 Hz, 2H, ArH), 8.01 (dd,
Reddish orange solid, 88% yield, mp 105 ꢁC; nmax (KBr): 1620
(C]O); 1H NMR (300 MHz, CDCl3):
d 2.41 (s, 3H, CH3), 5.05 (dd,

P. Singh et al. / European Journal of Medicinal Chemistry 45 (2010) 4968e4982
4977
Table 5
Percentage yields and melting points for indoleeoxindole conjugates 52e60.
CH3
N
Product
% yield
Mp (ꢁC)
CH3
N
N
O
R
NH
95
220
N
O
N
O
O
Cl
H
52
RCHO
MWI
Cl
61-65
66-70
N
N
82
81
210
Scheme 7. Synthesis of aromatic aldehyde-pyrazole conjugates.
53
54
N
Table 6
N
O
>240
Percentage yields and melting points for aromatic aldehydeepyrazole conjugates
66e70.
Reactant
R
Product
%
Mp
yield (ꢁC)
CH3
N
N
N
S
O
S
O
O
N
O
O
O
61
95
135
73
>250
Cl
66
CH3
CH3
55
CH3
N
N
N
O
N
62
94
192
N
N
O
85
87
81
148
238
Cl
56
67
CH3
N
N
N
O
N
O
O2N
O
O
63
96
158
O2N
57
Cl
68
N
Cl
N
O
Cl
>250
O
O
N
Cl
H3C
58
N
OH
64
90
200
O
OH
Cl
N
N
O
69
Cl
87
92
185
160
O
O
Cl
Cl
59
60
Cl
N
H3C
N
N
N
O
65
94
145
O
70

4978
P. Singh et al. / European Journal of Medicinal Chemistry 45 (2010) 4968e4982
4.1.14. 1,3-Dibenzyl-5-((1-benzyl-1H-indol-3-yl)methylene)
pyrimidine-2,4,6(1H,3H,5H)-trione (35)
Yellow solid, 80% yield, mp 192 ꢁC; nmax (KBr): 1630 (C]O);
R
1H NMR (300 MHz, CDCl3):
d
5.20 (s, 4H, 2ꢀCH2), 5.48 (s, 2H,
O
N
Cl
CH2), 7.18 (dd, J3 ¼ 7.2 Hz, J4 ¼ 1.8 Hz, 2H, ArH), 7.25e7.50 (m,
16H, ArH), 7.99 (d, J ¼ 7.8 Hz, 1H, ArH), 9.06 (s, 1H, ¼H), 9.69 (s,
O
N
Cl
Cl
1H, indole2-H); 13C NMR (normal/DEPT-135) (CDCl3):
d 44.5
Cl
RCHO
(ꢂve, CH2), 45.3 (ꢂve, CH2), 51.8 (ꢂve, CH2), 108.4 (C), 111.3 (þve,
CH), 112.4 (C), 118.8 (þve, CH), 123.3 (þve, CH), 124.2 (þve, CH),
126.8 (þve, CH), 127.4 (þve, CH), 127.5 (þve, CH), 128.3 (þve,
CH), 128.4 (þve, CH), 128.6 (þve, CH), 129.0 (þve, CH), 130.7 (C),
134.8 (C), 136.7 (C), 137.1 (C), 142.9 (þve, CH), 146.8 (þve, CH),
151.5 (C), 161.7 (C), 163.5 (C); FAB mass m/z 527 (M þ 2); (Found:
C 77.85, H 4.94, N 8.25; C34H27N3O3 requires C 77.70, H 5.18,
N 7.99).
NaHCO3, MWI
61-63, 65
71-74
Scheme 8. Synthesis of aromatic aldehyde-indolinone conjugates.
J2 ¼ 6 Hz, J3 ¼ 1.2 Hz, 2H, CH2), 6.31e6.56 (m, 1H, ¼H), 6.62 (s, 1H,
¼H), 7.09e7.20 (m, 1H, ArH), 7.21e7.38 (m, 4H, ArH), 7.47e7.49 (m,
3H, ArH), 7.81e7.90 (m, 3H, ArH), 7.95e7.98 (m, 1H, ArH), 7.98 (s,
1H, ¼H), 8.10 (t, J ¼ 1.8 Hz, 1H, ArH), 9.90 (s, 1H, indole2-H); 13C
4.1.15. (Z)-3-((1-Allyl-1H-indol-3-yl)methylene)indolin-2-one (36)
Yellow solid, 90% yield, mp 212 ꢁC; nmax (KBr): 1670 (C]O); 1H
NMR (normal/DEPT-135) (CDCl3):
d
12.8 (þve, CH3), 49.9 (ꢂve,
NMR (300 MHz, CDCl3):
d
4.8 (d, J ¼ 3 Hz, 2H, CH2), 5.2 (dd,
CH2), 109.5 (C), 111.3 (þve, CH), 118.1 (þve, CH), 118.7 (þve, CH),
120.8 (þve, CH), 122.7 (þve, CH), 122.8 (þve, CH), 123.8 (þve, CH),
124.1 (þve, CH), 126.4 (þve, CH), 126.6 (þve, CH), 128.2 (þve, CH),
128.6 (þve, CH), 129.7 (þve, CH), 129.9 (þve, CH), 133.9 (þve, CH),
135.5 (þve, CH),138.4 (þve, CH),140.7 (þve, CH),118.8 (C),119.4 (C),
129.3 (C),134.4 (C),134.6 (C),135.7 (C),136.7 (C),138.6 (C), 140.0 (C),
153.0 (C),161.5 (C),163.4 (C); FAB mass m/z 451 (Mþ), 451:453 (3:1);
(Found: C 74.62, H 4.91, N 9.68; C28H22ClN3O requires C 74.41,
H 4.91, N 9.30).
J2 ¼ 9 Hz, J3 ¼ 3 Hz, 2H, CH2), 6.02e6.09 (m, 1H, CH), 6.8 (d,
J ¼ 8.4 Hz, 1H, ArH), 7.0 (t, J ¼ 4.5 Hz, 1H, ArH), 7.1 (t, J ¼ 4.5 Hz, 1H,
ArH), 7.2e7.4 (m, 2H, ArH), 7.6 (d, J ¼ 4.5 Hz, 1H, ArH), 7.7e7.9 (m,
2H, ArH), 7.9 (s, 1H, ¼H), 9.4 (s, 1H, indole2-H); FAB mass m/z 301
(M þ 1); (Found: C 79.78, H 5.38, N 9.17; C20H16N2O requires
C 79.98, H 5.37, N 9.33).
4.1.16. (Z)-3-((1-(prop-2-ynyl)-1H-indol-3-yl)methylene)indolin-
2-one (37)
Dark red solid, 83% yield, mp 180 ꢁC; nmax (KBr): 1680 (C]O),
3450 (NH); 1H NMR (300 MHz, CDCl3):
d 2.4 (s, 1H, hH), 4.9 (d,
Table 7
J ¼ 2.4 Hz, 2H, CH2), 6.8e8.2 (m, 9H, ArHþ ¼H), 10.0 (s, 1H, indole2-
H); FAB mass m/z 298 (Mþ); (Found: C 80.58, H 4.68, N 9.37;
C20H14N2O requires C 80.52, H 4.73, N 9.39).
Percentage yields and melting points for aromatic aldehydeeindolinone conjugates
71e74.
Reactant
R
Product
%
Mp
yield (ꢁC)
4.1.17. (Z)-3-((1-Tosyl-1H-indol-3-yl)methylene)indolin-2-one (38)
Dark red solid, 76% yield, mp 232 ꢁC; nmax (KBr): 1668 (C]O),
3430 (NH); 1H NMR (300 MHz, CDCl3):
d 2.35 (s, 3H, CH3),
6.88e7.43 (m, 2H, ArH), 7.59 (t, J ¼ 8.4 Hz, 1H, ArH), 7.71e8.11 (m,
9H, ArH), 8.35 (s, 1H, ¼H), 9.74 (s, 1H, indole2-H); FAB mass m/z 414
(Mþ); (Found: C 69.58, H 4.68, N 6.37, S 7.80; C24H18N2O3S requires
C 69.55, H 4.38, N 6.76, S 7.74).
N
Cl
O
61
95
203
Cl
71
4.1.18. Compound 39
Yellow solid, 85% yield, mp >240 ꢁC; nmax (KBr): 1660 (C]O),
3432 (NH); 1H NMR (300 MHz, CDCl3):
d
6.8 (d, 1H, J ¼ 7.5 Hz, ArH),
N
N
Cl
7.04e7.24 (m, 4H, ArH), 7.45e7.48 (m, 2H, ArH), 7.59e7.66 (m, 3H,
ArH), 7.87 (t, J ¼ 6.6 Hz, 3H, ArH), 8.50e8.53 (m,1H, ArH), 9.54 (s,1H,
indole2-H); FAB mass m/z 364 (Mþ); (Found: C 79.20, H 4.52, N 7.76;
C24H16N2O2 requires C 79.11, H 4.43, N 7.69).
O
62
86
188
N
Cl
72
4.1.19. Compound 40
Yellow solid, 83% yield, mp > 240 ꢁC; nmax (KBr): 1680 (C]O),
Cl
3452 (NH); 1H NMR (300 MHz, CDCl3):
d
6.8 (d, J ¼ 7.5 Hz, 1H, ArH),
N
O2N
O
63
65
83
89
190
177
O2N
7.19e7.21 (m, 4H, ArH), 7.37e7.60 (m, 4H, ArH), 7.67e7.86 (m, 3H,
ArH), 8.04 (d, J ¼ 8.4 Hz, 1H, ArH), 9.54 (s, 1H, indole2-H); FAB mass
m/z 398 (Mþ), 398:400 (3:1); (Found: C 72.29, H 4.02, N 7.16;
C24H15ClN2O2 requires C 72.27, H 3.79, N 7.02).
Cl
73
Cl
4.1.20. Compound 41
Yellow solid, 80% yield, mp 196 ꢁC; nmax (KBr): 1704 (C]O), 3460
N
O
(NH);1H NMR(300 MHz, CDCl3):
d 6.75e6.84 (m, 2H, ArH), 7.07e7.71
Cl
(m, 9H, ArH), 7.78 (s, 1H, ¼H), 8.68 (d, J ¼ 7.5 Hz, 1H, indole2-H); FAB
mass m/z 433 (Mþ), 433:435:437 (9:6:1); (Found: C 66.57, H 3.32, N
6.56; C24H14Cl2N2O2 requires C 66.53, H 3.26, N 6.47).
74

P. Singh et al. / European Journal of Medicinal Chemistry 45 (2010) 4968e4982
4979
4.1.21. (3Z)-3-((1-Cinnamyl-1H-indol-3-yl)methylene)indolin-2-
one (42)
Dark yellow solid, 89% yield, mp 230 ꢁC; nmax (KBr): 1690 (C]O),
3430 (NH); 1H NMR (300 MHz, CDCl3):
5.01e5.04 (m, 2H, CH2),
6.36e6.40 (m, 1H, ¼H), 6.61 (d, J ¼ 6 Hz, 1H, ¼H), 6.88 (d, J ¼ 6.0 Hz,
1H, ArH), 7.06e7.64 (m, 10H, ArH), 7.79e7.81 (m, 2H, ArH), 7.96 (s,
1H, ¼H), 9.58 (s, 1H, indole2-H); FAB mass m/z 440 (Mþ); (Found: C
82.98, H 5.36, N 7.40; C26H20N2O requires C 82.95, H 5.35, N 7.44).
4.1.26. (Z)-3-((1-Benzyl-1H-indol-3-yl)methylene)-1-(2,6-
dichlorophenyl)indolin-2-one (47)
Dark yellow solid, 81% yield, mp 232 ꢁC; nmax (KBr): 1690 (C]O);
1H NMR (300 MHz, CDCl3):
d
d
5.37 (s, 2H, CH2), 6.48 (d, J ¼ 7.8 Hz,1H,
ArH), 7.16e7.55 (m, 13H, ArH), 7.76 (d, J ¼ 6.0 Hz, 1H, ArH), 7.95 (d,
J ¼ 7.8 Hz,1H, ArH), 8.13 (s,1H, ¼H), 9.61 (s,1H, indole2-H); 13C NMR
(normal/DEPT-135) (CDCl3):
d
51.2 (ꢂve, CH2), 96.1 (C), 108.6 (þve,
CH), 110.9 (þve, CH), 111.4 (C), 117.7 (C), 118.1 (þve, CH), 121.1 (þve,
CH),122.2 (þve, CH),123.0 (C),125.3 (C),126.6 (þve, CH),126.8 (þve,
CH), 127.8 (þve, CH), 127.9 (þve, CH), 128.8 (þve, CH), 128.9 (þve,
CH), 129.5 (C), 130.4 (þve, CH), 135.9 (C), 136.1 (C), 136.2 (C), 137.8
(þve, CH), 138.9 (C), 157.9 (C), 166.0 (C); FAB mass m/z 495 (Mþ),
495:497:499 (9:6:1); (Found: C 72.43, H 3.78, N 6.02; C30H20Cl2N2O
requires C 72.73, H 4.07, N 5.65).
4.1.22. (Z)-3-((1H-Indol-3-yl)methylene)-1-(2,6-dichlorophenyl)
indolin-2-one (43)
Dark yellow solid, 95% yield, mp 242 ꢁC; nmax (KBr): 1670 (C]O),
3450 (NH); 1H NMR (300 MHz, CDCl3):
d
6.48 (d, J ¼ 8.1 Hz, 1H,
ArH), 7.15e7.57 (m, 8H, ArH), 7.74 (d, J ¼ 8.7 Hz, 1H, ArH), 7.95 (d,
J ¼ 7.5 Hz, 1H, ArH), 8.14 (s, 1H, ¼H), 9.28 (bs, 1H, NH), 9.56 (d,
J ¼ 3 Hz, 1H, indole2-H); 13C NMR (normal/DEPT-135) (CDCl3):
4.1.27. Compound 48
d
110.6 (C), 111.1 (þve, CH), 118.2 (þve, CH), 119.0 (þve, CH), 119.0
Yellow solid, 90% yield, mp 170 ꢁC; nmax (KBr): 1690 (C]O); 1H
(þve, CH), 120.1 (þve, CH), 120.2 (C), 121.1 (þve, CH), 122.2 (þve,
CH), 126.1 (C), 127.3 (þve, CH), 127.9 (þve, CH), 127.9 (þve, CH),
128.0 (C), 128.9 (þve, CH), 129.4 (þve, CH), 129.4 (þve, CH), 130.8
(þve, CH), 135.5 (C), 139.7 (þve, CH), 140.0 (C), 144.9 (C), 159.3 (C);
FAB mass m/z 405 (M þ 1), 405:407:409 (9:6:1); (Found: C 67.95, H
3.30, N 7.12; C23H14Cl2N2O requires C 68.16, H 3.48, N 6.91).
NMR (300 MHz, CDCl3):
ArH), 8.01 (s, 1H, ¼H), 8.38e8.41 (m, 2H, ArH), 9.47 (s, 1H, indole2-
H); 13C NMR (normal/DEPT-135) (CDCl3):
108.9 (þve, CH), 109.2
d 6.38 (m, 1H, ArH), 7.13e7.92 (m, 13H,
d
(þve, CH), 116.4 (þve, CH), 117.8 (þve, CH), 118.9 (þve, CH), 120.1
(þve, CH), 122.4 (þve, CH), 123.1 (þve, CH), 124.3 (þve, CH), 124.6
(þve, CH), 125.1 (þve, CH), 125.4 (þve, CH), 126.0 (þve, CH), 127.6
(þve, CH),128.4 (þve, CH),128.71 (þve, CH),128.77 (þve, CH),128.8
(þve, CH), 128.9 (C), 129.1 (C), 129.9 (C), 130.4 (C), 130.6 (C), 132.4
(C), 132.7 (C), 133.9 (C), 135.7 (C), 136.0 (C), 165.0 (C), 168.8 (C); FAB
mass m/z 508 (Mþ), 508:510:512 (9:6:1); (Found: C 70.71, H 3.43,
N 5.63; C30H18Cl2N2O2 requires C 70.74, H 3.56, N 5.50).
4.1.23. (Z)-3-((1-Allyl-1H-indol-3-yl)methylene)-1-(2,6-
dichlorophenyl)indolin-2-one (44)
Dark yellow solid, 83% yield, mp 178 ꢁC; nmax (KBr): 1680 (C]
O); 1H NMR (300 MHz, CDCl3):
d
5.3 (ddd, J2 ¼ 16.8 Hz,
J3 ¼ 10.2 Hz, J4 ¼ 0.3 Hz, 2H, CH2), 6.03e6.12 (multiplet of 12 lines,
1H, CH), 6.45 (d, J ¼ 6.9 Hz, 2H, CH2), 7.00e7.55 (m, 9H, ArH), 7.82
(d, J ¼ 7.2 Hz, 1H, ArH), 7.91 (d, J ¼ 7.2 Hz, 1H, ArH), 7.95 (s, 1H,
¼H), 8.24 (s, 1H, indole2-H); 13C NMR (normal/DEPT-135) (CDCl3):
4.1.28. Compound 49
Yellow solid, 75% yield, mp >260 ꢁC; nmax (KBr): 1685 (C]O); 1H
NMR (300 MHz, CDCl3):
d
6.42 (d, J ¼ 7.5 Hz,1H, ArH), 6.43e7.99 (m,
d
49.3 (ꢂve, CH2), 108.9 (þve, CH), 110.2 (þve, CH), 111.5 (C), 118.0
14H, ArH), 8.35 (d, J ¼ 8.7 Hz, 1H, ¼H), 9.47 (s, 1H, indole2-H); FAB
mass m/z 543 (Mþ), 543:545:547:549 (27:27:9:1); (Found: C 66.00,
H 2.84, N 5.24; C30H17Cl3N2O2 requires C 66.26, H 3.15, N 5.15).
(C), 118.4 (þve, CH), 120.3 (þve, CH), 120.8 (C), 121.4 (þve, CH),
122.1 (ꢂve, CH2), 122.5 (C), 123.2 (þve, CH), 126.7 (C), 128.1 (þve,
CH), 128.3 (C), 128.9 (þve, CH), 130.2 (þve, CH), 130.4 (þve, CH),
130.9 (C), 131.1 (þve, CH), 132.3 (þve, CH), 135.8 (C), 136.5 (C), 141.1
(C), 167.9 (C); FAB mass m/z 445 (Mþ), 445:447:449 (9:6:1);
(Found: C 69.88, H 3.54, N 6.49; C26H18Cl2N2O requires C 70.12, H
4.07, N 6.29).
4.1.29. Compound 50
Yellow solid, 82% yield, mp 250 ꢁC; nmax (KBr): 1690 (C]O); 1H
NMR (300 MHz, CDCl3):
d
6.38 (d, J ¼ 7.8 Hz,1H, ArH), 7.08e7.70 (m,
11H, ArH), 7.82 (s, 1H, ¼H), 7.86e7.89 (m, 1H, ArH), 8.68e8.71 (m,
1H, ArH), 8.96 (s, 1H, indole2-H); FAB mass m/z 578 (Mþ)
576:578:580:582:584 (81:108:54:12:1); (Found: C 62.15, H 2.82, N
4.85; C30H16Cl4N2O2 requires C 62.31, H 2.79, N 4.84).
4.1.24. (Z)-1-(2,6-Dichlorophenyl)-3-((1-(prop-2-ynyl)-1H-indol-
3-yl)methylene)indolin-2-one (45)
Dark red solid, 85% yield, mp 168 ꢁC; nmax (KBr): 1678 (C]O); 1H
NMR (300 MHz, CDCl3):
d
2.56 (t, J ¼ 2.7 Hz, 1H, hH), 5.01 (d,
4.1.30. (3Z)-1-(2,6-Dichlorophenyl)-3-((1-cinnamyl-1H-indol-3-yl)
methylene)indolin-2-one (51)
J ¼ 2.4 Hz, 2H, CH2), 6.45 (d, J ¼ 7.5 Hz, 1H, ArH), 7.01e7.56 (m, 8H,
ArH), 7.82 (d, J ¼ 6.3 Hz, 1H, ArH), 7.94 (d, J ¼ 6.9 Hz, 1H, ArH), 8.11
(s, 1H, ¼H), 8.22 (s, 1H, indole2-H); 13C NMR (normal/DEPT-135)
Dark yellow solid, 91% yield, mp 190 ꢁC; nmax (KBr): 1700 (C]O);
1H NMR (300 MHz, CDCl3):
d 4.97 (multiplet of 8 lines, 2H, CH2),
(CDCl3):
d
36.5 (ꢂve, CH2), 75.0 (þve, CH), 77.1 (C), 108.7 (þve, CH),
6.28e6.44 (m, 1H, ¼H), 6.45e6.56 (m, 2H, ¼H þ ArH), 7.13e7.53 (m,
109.0 (C), 109.9 (þve, CH), 110.5 (C), 111.9 (C), 118.2 (C), 120.5 (þve,
CH), 121.5 (C), 121.7 (þve, CH), 122.2 (þve, CH), 122.4 (þve, CH),
123.5 (þve, CH), 128.3 (þve, CH), 128.4 (C), 128.9 (þve, CH), 129.8
(þve, CH),130.2 (þve, CH),130.5 (þve, CH),135.9 (C),136.1 (C),141.3
(C), 167.8 (C); FAB mass m/z 443 (Mþ), 443:445:447 (9:6:1);
(Found:. C 70.30, H 3.32, N 6.28; C26H16Cl2N2O requires C 70.44, H
3.64, N 6.32).
13H, ArH), 7.72e7.98 (m, 2H, ArH), 8.11 (s, 1H, ¼H), 9.53 (s, 1H,
indole2-H); 13C NMR (normal/DEPT-135) (CDCl3):
d
49.6 (ꢂve, CH2),
108.6 (þve, CH), 110.8 (þve, CH), 111.4 (C), 117.5 (C), 118.1 (þve, CH),
121.6 (þve, CH), 122.2 (þve, CH), 122.9 (þve, CH), 123.6 (þve, CH),
125.4 (C), 126.5 (þve, CH), 126.8 (þve, CH), 127.9 (þve, CH), 128.5
(þve, CH), 128.6 (þve, CH), 128.9 (þve, CH), 129.5 (C), 130.4 (þve,
CH), 133.1 (þve, CH), 131.2 (C), 135.9 (C), 136.0 (C), 136.2 (C), 137.3
(þve, CH), 138.9 (C), 166.0 (C); FAB mass m/z 521 (Mþ) 521:523:525
(9:6:1); (Found: C 73.50, H 4.52, N 5.56; C32H22Cl2N2O requires
C 73.71, H 4.25, N 5.37).
4.1.25. (Z)-1-(2,6-Dichlorophenyl)-3-((1-tosyl-1H-indol-3-yl)
methylene)indolin-2-one (46)
Yellow solid, 78% yield, mp >240 ꢁC; nmax (KBr): 1690 (C]O); 1H
NMR (300 MHz, CDCl3):
d
2.25 (s, 3H, CH3), 6.29 (d, J ¼ 7.5 Hz, 1H,
4.1.31. (Z)-3-((1H-indol-3-yl)methylene)indolin-2-one (52) [48,49]
Yellow solid, 95% yield, mp 220 ꢁC; nmax (KBr):1685 (C]O), 3470
d 6.82e6.92 (m, 1H, ArH),
6.97e7.32 (m, 3H, ArH), 7.41e7.47 (m, 1H, ArH), 7.58e7.93(m, 3H,
ArH), 7.98 (s, 1H, ¼H), 9.51 (s, 1H, indole2-H), 10.05 (s, 1H, NH);
ArH), 6.96e7.86 (m, 14H, ArH), 7.88 (s, 1H, ¼H), 9.66 (s, 1H, indole2-
H); FAB mass m/z 558 (M þ 1), 558:560:562 (9:6:1); (Found: C
64.55, H 3.88, N 5.12, S 6.06; C30H20Cl2N2O3S requires C 64.40, H
3.60, N 5.01, S 5.73).
(NH). 1H NMR (300 MHz, CDCl3):

4980
P. Singh et al. / European Journal of Medicinal Chemistry 45 (2010) 4968e4982
(Found: C 78.46, H 4.62, N 10.79; C32H22Cl2N2O requires C 78.44,
H 4.65, N 10.76).
4.1.36. Compound 57
Yellow solid, 87% yield, mp 238 ꢁC; nmax (KBr): 1699 (C]O); 1H
NMR (300 MHz, CDCl3): d 7.26e7.40 (m, 5H, ArH), 7.45e7.54 (m, 6H,
4.1.32. (Z)-1-Allyl-3-((1-allyl-1H-indol-3-yl)methylene)indolin-2-
one (53)
ArH), 7.60e7.82 (m, 6H, ArH), 7.88 (s, 1H, ¼H), 7.92 (d, J ¼ 6 Hz, 1H,
ArH), 9.22 (s, 1H, indole2-H); 13C NMR (normal/DEPT-135) (CDCl3):
Yellow solid, 82% yield, mp 210 ꢁC; nmax (KBr): 1680 (C]O); 1H
d
114.8 (C), 115.1 (þve, CH), 116.7 (þve, CH), 117.8 (þve, CH), 118.6
NMR (300 MHz, CDCl3):
d
4.48e4.51 (m, 2H, CH2), 4.83e4.85 (m,
(þve, CH), 122.9 (C), 124.5 (þve, CH), 124.6 (þve, CH), 125.0 (C),
125.7 (þve, CH), 126.2 (þve, CH), 127.9 (þve, CH), 128.1 (þve, CH),
128.4 (þve, CH), 128.8 (þve, CH), 128.9 (þve, CH), 129.2 (þve, CH),
129.3 (þve, CH), 129.4 (þve, CH), 129.8 (þve, CH), 130.2 (C), 133.1
(C), 132.0 (þve, CH), 132.4 (þve, CH), 134.7 (þve, CH), 134.9 (C),
135.7 (C), 138.2 (C); FAB mass m/z 468 (Mþ); (Found: C 79.19, H 4.31,
N 5.69; C31H20N2O3 requires C 79.47, H 4.30, N 5.98).
2H, CH2), 5.14e5.27 (m, 4H, 2ꢀCH2), 5.87e6.03 (m, 2H, 2ꢀCH), 6.84
(d, J ¼ 7.5 Hz, 1H, ArH), 7.05e7.40 (m, 5H, ArH), 7.64 (d, J ¼ 7.2 Hz,
1H, ArH), 7.91e7.94 (m, 1H, ArH), 7.99 (s, 1H, ¼H), 9.50 (s, 1H,
indole2-H); 13C NMR (normal/DEPT-135) (CDCl3):
d
42.0 (ꢂve, CH2),
49.7 (ꢂve, CH2), 77.2 (þve, CH), 108.3 (þve, CH), 110.6 (þve, CH),
111.2 (C), 116.9 (þve, CH), 117.7 (þve, CH), 118.0 (þve, CH), 118.2
(þve, CH), 118.3 (C), 121.4 (ꢂve, CH2), 122.7 (ꢂve, CH2), 125.1 (C),
126.7 (þve, CH), 126.8 (þve, CH), 129.3 (C), 132.0 (þve, CH), 132.2
(þve, CH), 136.1 (C), 136.8 (þve, CH), 139.9 (C), 166.9 (C); FAB mass
m/z 341 (M þ 1); (Found: C 81.11, H 5.74, N 8.19; C23H20N2O requires
C 81.15, H 5.92, N 8.23).
4.1.37. Compound 58
Yellow solid, 81% yield, mp >250 ꢁC; nmax (KBr): 1680 (C]O); 1H
NMR (300 MHz, CDCl3):
d 6.82e7.09 (m, 4H, ArH), 7.14e7.92 (m,
11H, ArH), 8.08 (s, 1H, ¼H), 9.50 (d, J ¼ 3.6 Hz, 1H, ArH), 10.04 (s, 1H,
indole2-H); FAB mass m/z 536 (Mþ), 536:538:540 (9:6:1); (Found:
C 69.15, H 3.64, N 5.51; C31H18Cl2N2O3 requires C 69.28, H 3.38,
N 5.21).
4.1.33. (Z)-1-(Prop-2-ynyl)-3-((1-(prop-2-ynyl)-1H-indol-3-yl)
methylene)indolin-2-one (54)
Dark red solid, 81% yield, mp >240 ꢁC; nmax (KBr): 1680 (C]O);
1H NMR (300 MHz, CDCl3):
d
1.57 (s, 1H, hH), 2.21 (s, 1H, hH), 4.66
4.1.38. Compound 59
(d, J ¼ 2.7 Hz, 2H, CH2), 5.74 (d, J ¼ 6.6 Hz, 2H, CH2), 7.04e7.35 (m,
5H, ArH), 7.63e7.70 (m, 2H, ArH), 7.88e7.91 (m, 1H, ArH), 7.93 (s,
1H, ¼H), 9.49 (s, 1H, indole2-H); 13C NMR (normal/DEPT-135)
Yellow solid, 87% yield, mp 185 ꢁC; nmax (KBr): 1690 (C]O); 1H
NMR (300 MHz, CDCl3): d 7.20e7.58 (m, 4H, ArH), 7.69e7.84 (m, 6H,
ArH), 8.39 (d, J ¼ 8.1 Hz, 2H, ArH), 8.62e8.65 (m, 4H, ArH); FAB mass
m/z 607 (M þ 1), 605:607:609:611:613 (81:108:54:12:1); (Found: C
61.46, H 2.34, N, 4.26; C31H16Cl4N2O3 requires C 61.41, H 2.66, N 4.62).
(CDCl3):
d
28.9 (ꢂve, CH2), 39.1 (ꢂve, CH2), 71.7 (C), 77.4 (þve, CH),
88.4 (þve, CH), 97.2 (þve, CH), 108.5 (þve, CH),111.1 (þve, CH),118.1
(þve, CH), 119.4 (C), 121.9 (þve, CH), 122.1 (þve, CH), 123.3 (þve,
CH), 126.2 (þve, CH), 127.2 (þve, CH), 133.9 (þve, CH), 139.1 (C),
203.7 (C); FAB mass m/z 337(M þ 1); (Found: C 81.94, H 4.59, N
8.25; C23H16N2O requires C 82.12, H 4.79, N 8.33).
4.1.39. (3Z)-1-Cinnamyl-3-((1-cinnamyl-1H-indol-3-yl)methylene)
indolin-2-one (60)
Yellow solid, 92% yield, mp 160 ꢁC; nmax (KBr): 1690 (C]O); 1H
NMR (300 MHz, CDCl3):
d
4.66 (d, J ¼ 4.8 Hz, 2H, CH2), 5.03 (d,
4.1.34. (Z)-1-Tosyl-3-((1-tosyl-1H-indol-3-yl)methylene)indolin-2-
one (55)
J ¼ 5.1 Hz, 2H, CH2), 6.22e6.39 (m, 2H, 2ꢀCH), 6.53e6.62 (m, 2H,
2ꢀCH), 6.92e7.47 (m, 16H, ArH), 7.66 (d, J ¼ 6.9 Hz, 1H, ArH),
7.93e7.96 (m, 1H, ArH), 8.02 (s, 1H, ¼H), 9.58 (s, 1H, indole2-H); 13C
Yellow solid, 73% yield, mp >250 ꢁC; nmax (KBr): 1663 (C]O); 1H
NMR (300 MHz, CDCl3):
d
2.36 (s, 3H, CH3), 2.43 (s, 3H, CH3), 7.21 (t,
NMR (normal/DEPT-135) (CDCl3):
d
41.7 (ꢂve, CH2), 49.5 (ꢂve,
J ¼ 6 Hz, 1H, ArH), 7.28e7.40 (m, 8H, ArH), 7.60 (d, J ¼ 7.4 Hz, 1H,
ArH), 7.71 (s, 1H, ¼H), 7.74 (d, J ¼ 7.6 Hz, 2H, ArH), 7.92 (d, J ¼ 6 Hz,
2H, ArH), 8.04 (d, J ¼ 8.2 Hz, 1H, ArH), 8.08 (d, J ¼ 8.3 Hz, 1H, ArH),
CH2), 108.4 (þve, CH), 110.7 (C), 111.3 (þve, CH), 117.8 (þve, CH),
118.1 (þve, CH), 118.5 (þve, CH), 121.4 (C), 122.9 (C), 123.6 (C), 125.2
(C), 126.4 (þve, CH), 126.8 (þve, CH), 127.6 (þve, CH), 128.0 (þve,
CH), 128.4 (þve, CH), 128.5 (þve, CH), 128.6 (þve, CH), 132.2 (þve,
CH), 133.1 (þve, CH), 136.2 (C), 136.4 (C), 136.8 (C); FAB mass m/z
492 (Mþ); (Found: C 85.27, H 5.85, N 5.59; C35H28N2O requires C
85.34, H 5.73, N 5.69).
9.52 (s, 1H, indole2-H); 13C NMR (normal/DEPT-135) (CDCl3):
d 21.6
(þve, CH3), 21.7 (þve, CH3), 113.7 (þve, CH), 113.8 (þve, CH), 115.1
(C), 118.2 (þve, CH), 118.7 (þve, CH), 121.9 (C), 123.9 (þve, CH), 124.2
(þve, CH), 124.5 (C), 125.3 (þve, CH), 125.7 (þve, CH), 127.2 (þve,
CH), 127.9 (þve, CH), 129.2 (þve, CH), 129.7 (þve, CH), 130.1 (þve,
CH),130.2 (C),132.9 (C),134.3 (C),134.7 (C),135.6 (C),137.3 (C),145.4
(C), 145.5 (C); FAB mass m/z 521 (Mþ); (Found: C 65.43, H 3.94, N
4.49, S 10.93; C31H24N2O5S2 requires C 65.47, H 4.25, N 4.93, S 11.28).
4.1.40. (4Z)-1-(3-Chlorophenyl)-3-methyl-4-((E)-3-
phenylallylidene)-1H-pyrazol-5(4H)-one (66)
Reddish orange solid, 95% yield, mp 135 ꢁC; nmax (KBr): 1753
(C]O); 1H NMR (300 MHz, CDCl3):
d 2.35 (s, 3H, CH3), 7.11e7.14 (m,
4.1.35. (Z)-1-Benzyl-3-((1-benzyl-1H-indol-3-yl)methylene)
indolin-2-one (56)
1H, ArH), 7.15e7.44 (m, 6H, ArH), 7.64e7.67 (m, 2H, ArH), 7.89e7.93
(m, 1H, ArH), 8.04 (t, J ¼ 2.1 Hz, 1H, ArH), 8.54 (dd, J ¼ 15.4 Hz,
Yellow solid, 85% yield, mp 148 ꢁC; nmax (KBr): 1680 (C]O); 1H
J ¼ 11.4 Hz, 1H, CH); 13C NMR (normal/DEPT-135) (CDCl3):
d 12.7
NMR (300 MHz, CDCl3):
d
5.09 (s, 2H, CH2), 5.47 (s, 2H, CH2),
(þve, CH3), 116.2 (þve, CH), 118.3 (þve, CH), 123.1 (þve, CH), 124.4
(þve, CH), 125.3 (C), 128.3 (þve, CH), 128.6 (þve, CH), 129.0 (þve,
CH), 129.7 (þve, CH), 130.9 (þve, CH), 134.5 (C), 135.4 (C), 139.4 (C),
144.9 (þve, CH), 149.8 (þve, CH), 149.9 (C), 163.1 (C); FAB mass m/z
323 (M þ 1) 323:325 (3:1); (Found: C 70.71, H 4.70, N 8.39;
C19H15ClN2O requires C 70.70, H 4.68, N 8.68).
6.73e6.87 (m, 3H, ArH), 7.07e7.40 (m, 11H, ArH), 7.66e7.86 (m, 3H,
ArH), 7.97 (d, J ¼ 6 Hz, 1H, ArH), 8.06 (s, 1H, ¼H), 9.68 (s, 1H,
indole2-H); 13C NMR (normal/DEPT-135) (CDCl3):
d
43.6 (ꢂve, CH2),
51.1 (ꢂve, CH2), 108.6 (þve, CH), 110.9 (þve, CH), 111.4 (C), 117.8
(þve, CH), 118.6 (C), 118.1 (þve, CH), 121.51 (þve, CH), 121.55 (þve,
CH), 123.0 (þve, CH), 125.2 (C), 126.71 (þve, CH), 126.73 (þve, CH),
126.8 (þve, CH), 126.9 (þve, CH), 127.1 (þve, CH), 127.2 (þve, CH),
127.3 (þve, CH), 127.9 (þve, CH), 128.71 (þve, CH), 128.75 (þve, CH),
128.92 (þve, CH), 128.94 (þve, CH), 129.5 (C), 136.31 (C), 136.36 (C),
136.68 (C), 137.3 (þve, CH), 140.0 (C), 167.3 (C); FAB mass m/z 440
(Mþ); (Found: C 84.42, H 5.49, N 6.25; C31H24N2O requires C 84.52,
H 5.49, N 6.36).
4.1.41. (4Z)-1-(3-Chlorophenyl)-3-methyl-4-((pyridin-2-yl)
methylene)-1H-pyrazol-5(4H)-one (67)
Reddish orange solid, 94% yield, mp 192 ꢁC; nmax (KBr): 1650
(C]O); 1H NMR (300 MHz, CDCl3):
d 1.71 (s, 3H, CH3), 7.11e7.19 (m,
2H, ArH), 7.24e7.35 (m, 3H, ArH), 7.58e7.64 (m, 1H, ArH), 7.72e7.76
(m, 1H, ArH), 7.90 (t, J ¼ 2.1 Hz, 1H, ArH), 8.50e8.52 (m, 1H, ArH);

P. Singh et al. / European Journal of Medicinal Chemistry 45 (2010) 4968e4982
4981
13C NMR (normal/DEPT-135) (CDCl3):
d
11.8 (þve, CH3), 48.5 (þve,
1H, ArH), 7.13e7.54 (m, 5H, ArH), 7.84e7.93 (m, 3H, ArH), 8.35 (d,
J ¼ 8.7 Hz, 2H, ArH); FAB mass m/z 411 (Mþ) 411:413:415 (9:6:1);
(Found: C 61.36, H 2.83, N 6.68; C21H12Cl2N2O3 requires C 61.33, H
2.94, N 6.81).
CH), 98.3 (C), 118.5 (þve, CH), 120.7 (þve, CH), 122.9 (þve, CH),
124.7 (þve, CH), 125.1 (þve, CH), 129.7 (þve, CH), 134.4 (C), 138.6
(þve, CH), 140.1 (C), 146.5 (þve, CH), 147.5 (C), 152.5 (C), 162.3 (C);
FAB mass m/z 297 (M þ 1) 297:299 (3:1); (Found: C 64.21, H 4.06,
N 13.84; C16H12ClN3O requires C 64.54, H 4.06, N 14.11).
4.1.48. (Z)-3-((Anthracen-10-yl)methylene)-1-(2,6-dichlorophenyl)
indolin-2-one (74)
4.1.42. (4Z)-4-(4-Nitrobenzylidene)-1-(3-chlorophenyl)-3-methyl-
1H-pyrazol-5(4H)-one (68)
Shiny yellowsolid, 89% yield, mp 177 ꢁC; nmax (KBr): 1680 (C]O);
1H NMR (300 MHz, CDCl3):
J ¼ 7.5 Hz, 1H, ArH), 6.47e6.53 (m, 1H, ArH), 7.01e7.24 (m, 1H, ArH),
7.37 (s,1H, ¼H), 7.39e7.57 (m, 6H, ArH), 8.05e8.13 (m, 4H, ArH), 8.57
(s, 1H, ArH), 8.71 (s, 1H, ArH); 13C NMR (normal/DEPT-135) (CDCl3):
d
6.04 (d, J ¼ 7.5 Hz, 1H, ArH), 6.37 (d,
Reddish orange solid, 96% yield, mp 158 ꢁC; nmax (KBr): 1650
(C]O); 1H NMR (300 MHz, CDCl3):
d 1.81 (s, 3H, CH3), 6.69e6.72
(m, 1H, ArH), 7.26e7.34 (m, 3H, ArH), 7.51e7.73 (m, 2H, ArH),
8.08e8.15 (m, 2H, ArH), 8.48 (d, J ¼ 8.7 Hz, 1H, ArH); FAB mass m/z
341 (Mþ) 341:343 (3:1); (Found: C 59.63, H 3.31, N 12.16;
C17H12ClN3O3 requires C 59.75, H 3.54, N 12.30).
d
108.9 (þve, CH), 121.1 (C), 122.6 (þve, CH), 124.4 (þve, CH), 125.6
(þve, CH), 125.7 (þve, CH), 126.6 (þve, CH), 128.3 (C), 128.5 (þve,
CH),128.91 (þve, CH),128.95 (C),129.0 (C),129.6 (C),130.5 (C),130.6
(C), 130.7 (þve, CH), 131.2 (C), 134.5 (þve, CH), 135.8 (C), 142.3 (C),
166.2 (C); FAB mass m/z 466 (Mþ) 466:468:470 (9:6:1); (Found: C
74.87, H 3.53, N 3.09; C29H17Cl2NO requires C 74.69, H 3.67, N 3.00).
4.1.43. (4Z)-1-(3-Chlorophenyl)-4-((2-hydroxynaphthalen-3-yl)
methylene)-3-methyl-1H-pyrazol-5(4H)-one (69)
Orange solid, 90% yield, mp 200 ꢁC; nmax (KBr): 1685 (C]O); 1H
NMR (300 MHz, CDCl3): d 2.38 (s, 3H, CH3), 7.23e7.46 (m, 6H, ArH),
4.2. In-vitro tumor growth inhibitory activities
7.62e7.79 (m, 2H, ArH), 7.82e7.88 (m, 2H, ArH), 7.97e8.00 (m, 1H,
ArH); FAB mass m/z 362 (Mþ) 362:364 (3:1); (Found: C 69.43, H
4.11, N 7.66; C21H15ClN2O2 requires C 69.52, H 4.17, N 7.72).
In-vitro tumor growth inhibitory activities of these compounds
were investigated at NCI, Bethesda on 60 cell line panel of human
cancer cells using standard procedure. Compounds were first tested
at 10ꢂ5 M for the growth inhibitory activities at entire 60 cell line
panel. Compounds with considerable activities at 10ꢂ5 M concen-
tration (as per the standard of NCI) were subjected to detailed
tumor growth inhibitory studies at five concentrations viz. 10ꢂ4 M,
10ꢂ5 M, 10ꢂ6 M, 10ꢂ7 M and 10ꢂ8 M.
4.1.44. (4Z)-4-((Anthracen-10-yl)methylene)-1-(3-chlorophenyl)-
3-methyl-1H-pyrazol-5(4H)-one (70)
Reddish orange solid, 94% yield, mp 145 ꢁC; nmax (KBr): 1690
(C]O); 1H NMR (300 MHz, CDCl3):
d 2.59 (s, 3H, CH3), 7.04e7.07
(m, 1H, ArH), 7.17e7.7.40 (m, 1H, ArH), 7.50e7.58 (m, 5H, ArH),
7.76e7.79 (m, 1H, ArH), 7.92e8.09 (m, 4H, ArH), 8.58 (d, J ¼ 3.6 Hz,
1H, ArH), 8.78 (s, 1H, CH); FAB mass m/z 396 (Mþ) 396:398 (3:1);
(Found: C 75.33, H 4.31, N 7.06; C25H17ClN2O requires C 75.66, H
4.32, N 7.06).
4.3. Maximum tolerable dose test (acute toxicity description)
A single mouse was given a single injection [Intravenous
injection (IV)] of 400 mg/kg; a second mouse received a dose of
200 mg/kg and a third mouse received a single dose of 100 mg/kg.
Dose volumes were generally 0.1 ml/10 gm body weight. The mice
were observed for a period of 2 weeks. They were sacrificed if they
lost more than 20% of their body weight or if there were other signs
of significant toxicity. If all 3 mice were sacrificed, then the next 3
dose levels (50, 25, 12.5 mg/kg) were tested in a similar way. The
process was repeated until a tolerated dose was found. The
maximum tolerated dose was used to calculate the amount of
material given to experimental mice during antitumor testing.
4.1.45. (3Z)-1-(2,6-Dichlorophenyl)-3-((E)-3-phenylallylidene)
indolin-2-one (71)
Shiny yellow solid, 95% yield, mp 203 ꢁC; nmax (KBr):1700 (C]O);
1H NMR (300 MHz, CDCl3):
d
6.42e6.45 (m, 1H, ¼H), 7.12e7.25 (m,
4H, ArH), 7.34e7.46 (m, 5H, 4ꢀ ArHþ, ¼H), 7.49e7.84 (m, 5H, ArH);
13C NMR (normal/DEPT-135) (CDCl3):
d
109.2 (þve, CH), 122.4 (þve,
CH),123.2 (þve, CH),123.7 (þve, CH),124.4 (C),127.7 (þve, CH),128.8
(þve, CH), 128.91(þve, CH), 128.94 (þve, CH), 129.7 (þve, CH), 130.6
(þve, CH), 135.71 (C), 135.73 (C), 135.9 (C), 136.9 (þve, CH), 141.5 (C),
144.8 (þve, CH), 167.1(C); FAB mass m/z 392 (Mþ) 392:394:396
(9:6:1); (Found: C 70.30, H 3.56, N 3.63; C23H15Cl2NO requires C
70.42, H 3.85, N 3.37).
4.4. In-vivo anticancer activities
4.1.46. (Z)-1-(2,6-Dichlorophenyl)-3-((pyridin-2-yl)methylene)
indolin-2-one (72)
A panel of 12 tumor cell lines viz. NCI-H23, NCI-H522, MDA-MB-
231, MDA-MB-435, SW-620, COLO 205, LOX, UACC-62, OVCAR-3,
OVCAR-5, U251 and SF-295, cultivated in RPMI-1640 containing
10% FBS and 2 mM glutamine was used. The cell suspension
(2e10 ꢀ 106 cells/ml) was flushed into 1 mm (internal diameter)
polyvinylidene fluoride hollow fibers with molecular weight
exclusion of 500,000 Da. The hollow fibers were heat-sealed at
2 cm intervals and the samples generated from these seals were
placed into tissue culture medium and incubated at 37 ꢁC in 5% CO2
for 24e48 h prior to implantation. Samples of each tumor cell line
preparation were quantitated for viable cell mass by a stable
endpoint MTT (3-[4,5-dimethylthiazole-2-yl]-2,5-diphenylte-
trazolium bromide) assay before and after the administration of
test agent. The optical density of each sample was determined
spectrophotometrically at 540 nm and the mean of each treatment
group was calculated. A 50% or greater reduction in percent net
growth in the treated samples compared to the vehicle control
samples was considered a positive result.
Shiny yellow solid, 86%, mp 188 ꢁC; nmax (KBr): 1668 (C]O); 1H
NMR (300 MHz, CDCl3):
d
6.41 (d, J ¼ 7.6 Hz, 1H, ArH), 7.13 (t,
J ¼ 7.6 Hz, 1H, ArH), 7.15e7.41 (m, 3H, ArH), 7.53 (d, J ¼ 8.1 Hz, 2H,
ArH), 7.66 (d, J ¼ 7.7 Hz, 1H, ArH), 7.82e7.85 (m, 2H, ArH), 8.91 (d,
J ¼ 4.5 Hz, 1H, ArH), 9.12 (d, J ¼ 7.6 Hz, 1H, ArH); 13C NMR (normal/
DEPT-135) (CDCl3):
d
109.2 (þve, CH), 122.4 (þve, CH), 123.2 (þve,
CH), 123.7 (þve, CH), 124.4 (C), 127.7 (þve, CH), 128.8 (þve, CH),
128.91 (þve, CH), 128.94 (þve, CH), 129.7 (þve, CH), 130.6 (þve, CH),
135.71 (C),135.73 (C),135.9 (C),136.9 (þve, CH),141.5 (C),144.8 (þve,
CH),167.1(C); FAB mass m/z 366 (Mþ) 366:368:370 (9:6:1); (Found: C
65.15, H 3.28, N 7.38; C20H12Cl2N2O requires C 65.41, H 3.29, N 7.63).
4.1.47. (Z)-3-(4-Nitrobenzylidene)-1-(2,6-dichlorophenyl)indolin-
2-one (73)
Yellow solid, 83% yield, mp 190 ꢁC; nmax (KBr): 1706 (C]O); 1H
NMR (300 MHz, CDCl3):
d
6.41 (d, J ¼ 7.8 Hz,1H, ArH), 6.86e6.97 (m,

4982
P. Singh et al. / European Journal of Medicinal Chemistry 45 (2010) 4968e4982
[24] NCI data base.www.dtp.nci.nih.gov NSC 123127.
Acknowledgment
[25] R.E. Wallace, K.C. Murdock, R.B. Angier, F.E. Durr, Cancer Res. 39 (1979)
1570e1574.
We thank Council of Scientific and Industrial Research, New
Delhi for the research grant. MK thanks University Grants
Commission New Delhi for fellowship (under SAP). We also thank
CDRI, Lucknow for recording mass spectra. NCI, USA is gratefully
acknowledged for investigating antitumor activities.
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Appendix. Supplementary data
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