(E)-3-(Aryl(arylamino)methylene)indolin-2-one derivatives: An efficient synthetic approach and evaluation of their cancer inhibitory activity

Article abstract of DOI:10.3184/174751918X15166485941737

A series of (E)-3-(aryl(arylamino)methylene)indolin-2-one derivatives were synthesised using an efficient synthetic approach. The method involved reaction of 3-bromo-3-(bromo(aryl)methyl)indolin-2-one with substituted anilines through nucleophilic substitution and a simultaneous elimination using NaHCO₃ in DMF. The anticancer activity of the products against four cell lines, HCT-116, A549, SKOV3 and MDA-MB-231, was also evaluated, and several compounds showed moderate inhibitory activity.

Full text of DOI:10.3184/174751918X15166485941737

44 RESEARCH PAPER
VOL. 42 JANUARY, 44–49
JOURNAL OF CHEMICAL RESEARCH 2018
(E)-3-(Aryl(arylamino)methylene)indolin-2-one derivatives: an efficient
synthetic approach and evaluation of their cancer inhibitory activity
Hongwu Jianga, Zhiyuan Fenga, Taiping Chena, Zicheng Lia*, Wencai Huanga, Youfu Luob
and Yinglan Zhaob
aSchool of Chemical Engineering, Sichuan University, Chengdu 610065, Sichuan, P.R. China
bState Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Chengdu 610041, Sichuan, P.R. China
A series of (E)-3-(aryl(arylamino)methylene)indolin-2-one derivatives were synthesised using an efficient synthetic approach. The
method involved reaction of 3-bromo-3-(bromo(aryl)methyl)indolin-2-one with substituted anilines through nucleophilic substitution and
a simultaneous elimination using NaHCO3 in DMF. The anticancer activity of the products against four cell lines, HCT-116, A549, SKOV3
and MDA-MB-231, was also evaluated, and several compounds showed moderate inhibitory activity.
Keywords: indolin-2-one derivative, nucleophilic substitution reaction, anticancer activity
Indolin-2-one fragments can be found in many natural plants,
for example, horsfiline from the traditional herb Horsfieldia
kingii1, and indirubin from Indigo naturalis2,3, which show
diverse biological activities, such as anti-angiogenesis,4 anti-
inflammatory,5 antitumour6 and anti-HIV7. Their derivatives
have attracted a lot of attention, with their significant anticancer
activity, as multi-target drugs.8 Li et al.9 reported that the
indolin-2-one derivatives can bind with several targets related
to the proliferation of cancer cells that are epidermal growth
factor receptors (EGFR), fibroblast growth factor receptors
(FGFR), vascular endothelial growth factor receptors (VEGFR)
and platelet-derived growth factor receptors (PDGFR). In
vivo, they can inhibit the corresponding receptor to block the
growth of cancer cells.10 Additionally, some of them have now
come onto the market, including Sunitinib,11 Vinflunine,12
Osimertinib13 and Nintedanib (see Fig. 1).14
Based on an indolin-2-one core and the precedent of the
newly marketed drug Nintedanib, which has been applied to
cure the symptoms of idiopathic pulmonary fibrosis (IPF)15
with good efficacy, we designed and synthesised a series
of compounds containing the indolin-2-one structure. A
general method for synthesising these compounds includes
the condensation reaction of indolin-2-one and trimethyl
orthobenzoate, which is expensive.16–18 Cantagrel et al.19
reported another synthetic method using the reaction of
3-(chloro(phenyl)methylene)-indolin-2-one with aliphatic
amines.
This paper reports a novel approach to synthesising (E)-3-
(aryl(arylamino) methylene)indolin-2-one derivatives by reacting
3-bromo-3-(bromo(phenyl)methyl)-indolin-2-one
derivatives
with aniline derivatives through a simultaneous nucleophilic
substitution and elimination reaction in one step using NaHCO3 in
DMF, which is easy to handle, with moderate to high yields. Their
antitumour activities against HCT-116, A549, SKOV3 and MDA-
MB-231 were evaluated and their preliminary structure–activity
relationships are also discussed.
Results and discussion
Chemistry
The preparation of (E)-3-(aryl(arylamino)methylene)indolin-
2-one derivatives is illustrated in Schemes 1–3. An acyl
chloride obtained from compound 1 (prepared according to the
literature20–22) and SOCl2, reacted with morpholine, piperidine and
N-methylpiperazine to give compounds 2a–c, which were then
reduced to compounds 3a–c under H2/Pd-C.
Compounds 4a–d were reacted with benzaldehyde to give 5a–d
in the presence of piperidine in ethanol. Compounds 6a–d were
then prepared in high yields by adding bromine in dichloromethane
to solutions of 5a–d in CH2Cl2 (Scheme 2).
Compounds 6a–d were reacted with compounds 7a–c or 3a–c
in DMF in the presence of NaHCO3 to form the target compounds
8a–e or 9a–g, respectively (Scheme 3). The use of K2CO3 as base
in DMF was also studied, but gave either more impurities and
lower yields or even no product at all. Dibromo-compound 6 was
t
F
F
NE 2
O
N
H
N
N
N
F
H
OCOMe
COOMe
H
H
H
N
H
O
N
MeOOC
N
H
O
O
Sunitinib
Vinflunine
O
N
N
N
N
O
NH
N
NH
N
N
O
N
N
H
O
N
O
H
O
Nintedanib
Osimertinib
Fig. 1 Several indole derivatives as antitumour drugs.
* Correspondent. E-mail: sculzc@scu.edu.cn

JOURNAL OF CHEMICAL RESEARCH 2018 45
O2N
.
H2N
O2N
1
l
SOC 2
X
X
H2, Pd-C
.
N
2
NH NEt3
N
OCH2COOH
O
O
X
O
O
1
2a–c
a:x=O; b:x=CH2; c;x=N-CH3
3a–c
Scheme 1 Synthetic method for 2-(4-aminophenoxy)acetamide derivatives 3a–c.
Br
4–6a: R1=H
CHO
R1
R1
Br
R1
4–6b: R1=CH3
4–6c: R1=Cl
4–6d: R1=Br
Br2
O
O
O
piperidine
N
H
CH2Cl2
N
H
N
H
4a–d
5a–d
6a–d
Scheme 2 Synthetic pathway for 3-bromo-3-(bromo(phenylmethyl))-indolin-2-one derivatives 6a–d.
R2
H2N
R2
HN
7a–c
7a: R2=H, 7b: R2=F, 7c: R2=CH3
8a: R1=Cl, R2=H
NaHCO3/DMF
R1
8b: R1=CH3, R2=CH3
8c: R1=CH3, R2=F
8d: R1=Br, R2=CH3
8e: R1=Br, R2=F
O
O
N
+
6a–d
H
8a–e
X
9a: R1=H, X=O
O
N
9b: R1=Cl, X=O
9c: R1=Br, X=O
9d: R1=H, X=CH2
9e: R1=Br, X=CH2
9f: R1=H, X=N-CH3
9g: R1=Br, X=N-CH3
3a–c
NaHCO3/DMF
HN
R1
O
N
H
9a–g
Scheme 3 Synthetic pathway for target compounds 8a–e and 9a–g.
first transformed into a vinyl bromide derivative, which was then
reacted with 3 or 7 very slowly, but no product was obtained. The
reaction proceeded very slowly due to the p–π conjugated system
of the bromine atom and the C=C double bond for the intermediate
of the 3-(bromo(phenyl)methylene)-indolin-2-one derivatives. As
for compound 6, nucleophilic substitution happened more easily
and faster between the bromoalkane with aniline, and then the
elimination reaction proceeded to form the target compound.
In the H–H COSY NMR spectrum of compound 5b, the
signal of H4 at 7.35 ppm interacted with the signal of H2′ at
7.72 ppm and that of H6′ at the phenyl group, indicating there
existed an E configuration. Intermediate 6 has different possible
stereoisomers that can be seen in the proton-NMR spectra,
with signals of the NH of the indolinone and the CHBr double
singlets. However, the existence of isomers does not affect the
following reaction, as all raw materials are transferred to the
product during the reaction. The proton-NMR signals of H4
in the indolinones of compounds 8 and 9 are located at greater
than 7, according to the literature.19 If their structures have the Z
configuration, H4 will correlate with H2′ and H6′ of the phenyl
group through NOE and the signal of H4 will be lower than 7.
In the H–H COSY NMR of compound 8d, the signal of H4
at 7.85 ppm interacted with the signal of H3′ at 6.58 ppm and
that of H5′ of the anilino group, although the intramolecular
hydrogen bond that can exist between the amino group
and the carbonyl group will promote the formation of the
Z configuration, the signal of H4 of our target compounds
occurred at about 7.6 ppm and, as noted in the literature,16 the
signals of H4 of similar analogues are located at 5.7–5.9 ppm.
Hence, we concluded that the E configuration products were
formed during the reaction. All of the target compounds were
easily purified by recrystallisation, and their structures were
confirmed by 1H NMR, 13C NMR and ESI-MS.
Biological evaluation
The antitumour activity in vitro of compounds 8a–e and 9a–g
was evaluated by the MTT assay and compared with that of
the reference drug gefitinib. Four different cancer cell lines,

46 JOURNAL OF CHEMICAL RESEARCH 2018
Table 1. IC50 values of target compounds 8a–e and 9a–g against tumour
cells in vitro
literature, and the main intermediates were characterised by IR,
1H NMR, 13C NMR and mass spectra.
IC50a,b (μM)
A549
>50
>50
>50
16.4
15.8
>50
>50
>50
23.3
46.6
>50
NDc
12.6
Compound
Synthesis of N-2-(4-nitrophenoxy)acetamide derivatives (2a–c);
general procedure
HCT116
>50
>50
14.5
23.3
13.6
>50
SKOV3
>50
>50
>50
22.4
24.8
>50
>50
>50
>50
>50
>50
>50
>50
MDA-MB-231
>50
>50
>50
15.2
30.3
34.6
>50
>50
8a
8b
8c
8d
8e
9a
9b
9c
9d
9e
9f
A mixture of compound 1 (10 mmol) and three drops of DMF in
thionyl chloride (10 mL) was refluxed for 1.5 h. Thionyl chloride
was evaporated under vacuum and CH2Cl2 (20 mL) was added.
To a mixture of the heterocyclic aliphatic amine (12 mmol) and
triethylamine (30 mmol) in CH2Cl2 (50 mL) the acyl chloride solution
was slowly added dropwise, with constant stirring and cooling of the
reaction mixture using an ice-water bath. The mixture was then stirred
for 30 min at room temperature, the organic layer was separated,
washed twice with salt water, dried with anhydrous sodium sulfate,
and the solvent was removed to give the crude product, which was
recrystallised from ethanol to give compound 2.
>50
>50
20.5
16.8
>50
36.2
19.4
23.0
18.8
42.1
28.2
1-(Morpholin-4-yl)-2-(4-nitrophenoxy)ethanone (2a): Yellow solid;
9g
TNd
1
yield 89%; m.p. 153–154 °C; H NMR (400 MHz, DMSO-d6): δ 3.45
32.2
(s, 4H, CH2), 3.61 (d, J = 21.4 Hz, 4H, CH2), 5.08 (s, 2H, CH2), 7.13 (d,
J = 9.2 Hz, 2H, ArH), 8.20 (d, J = 9.2, 2H, ArH); ESI-MS m/z: 267.4
(M+H).
2-(4-Nitrophenoxy1)-1-(piperidin-1-yl)ethanone (2b): Yellow solid;
yield 88%; m.p. 60–62 °C; 1H NMR (400 MHz, DMSO-d6): δ 1.44 (s,
2H, CH2), 1.58 (dd, J = 12.5, 7.2 Hz, 4H, CH2), 3.34–3.48 (m, 4H, CH2),
5.04 (s, 2H, CH2), 7.09 (d, J = 9.2 Hz, 2H, ArH), 8.19 (d, J = 9.2, 2H,
ArH); ESI-MS m/z: 265.2 (M+H).
aIC50 values represent the concentration that causes 50% growth inhibition.
bIC50 values were the mean values of three repeated experiments.
cNot determined.
dTN is gefitinib.
human colonic cancer cell (HCT116), human lung cell (A549),
human ovarian cancer cell (SKOV3) and human breast cancer
cell (MAD-MB-231), were chosen to be tested. The results are
presented in Table 1.
1-(4-Methylpiperazin-1-yl)-2-(4-nitrophenoxy)ethanone (2c): Yellow
1
solid; yield 84%; m.p. 213–214 °C; H NMR (400 MHz, DMSO-d6): δ
2.19 (s, 3H, CH3), 2.28 (d, J = 4.5 Hz, 2H, CH2), 2.35 (s, 2H, CH2), 3.44
(q, J = 5.2 Hz, 4H, CH2), 5.06 (s, 2H, CH2), 7.11 (d, J = 9.2 Hz, 2H, ArH),
8.20 (d, J = 9.2 Hz, 2H, ArH); ESI-MS m/z: 280.1 (M+H).
As can be seen from the table, 8d and 8e showed good
inhibitory activity against all four cells, 8c and 9e displayed
better activity than gefitinib towards HCT116, 9d displayed the
same activity as gefitinib towards HCT116, A549 and MDA-
MB-231, 8e, 9a, 9e and 9g showed the same or better activity
than gefitinib against MAD-MB-231, and only 8d and 8e showed
moderate inhibitory activity against SKOV3. Other compounds
showed either low or no inhibitory activity. It appears that the
bromo group at position 5 of indolin-2-one promoted activity
in compounds 8a–e, and compounds containing the piperidine
group could increase the activity in compounds 9a–g.
Synthesis of 2-(4-aminophenoxy)acetamide derivatives (3a–c);
general procedure
Compound 2 (10 mmol) in ethanol (50 mL) was hydrogenated at
0.2–0.4 MPa, catalysed by 10% Pd-C at room temperature, and
recrystallised from ethanol to yield compound 3.
2-(4-Aminophenoxy)-1-morpholinoethanone (3a): Pink solid;
1
yield 83%; m.p. 138–139 °C (lit.23 m.p.138 °C); H NMR (400 MHz,
DMSO-d6): δ 3.39 (d, J = 4.9 Hz, 4H, CH2), 3.56 (s, 4H, CH2), 4.61
(s, 2H, CH2), 4.64 (s, 2H, NH2), 6.49 (d, J = 8.4 Hz, 2H, ArH), 6.65 (d,
J = 8.1 Hz, 2H, ArH).
2-(4-Aminephenoxy)-1-(piperidin-1-yl)ethanone (3b): Oily (lit.23
oily); yield 79%; 1H NMR (400 MHz, DMSO-d6): δ 1.40 (s, 2H, CH2),
1.46–1.61 (m, 4H, CH2), 3.28–3.42 (m, 4H, CH2), 4.59 (s, 2H, CH2),
4.62 (s, 2H, NH2), 6.50 (d, J = 8.2 Hz, 2H, ArH), 6.65 (d, J = 8.2 Hz,
2H, ArH).
2-(4-Aminophenoxy)-1-(4-methylpiperazin-1-yl)ethanone (3c): Yellow
solid; yield 78%; m.p. 72–73 °C (lit.23 73 °C); 1H NMR (400 MHz,
DMSO-d6): δ 2.15 (s, 3H, CH3), 2.22 (d, J = 4.5 Hz, 2H, CH2), 2.30 (s, 2H,
CH2), 3.26–3.44 (m, 4H, CH2), 4.62 (s, 2H, CH2), 4.66 (s, 2H, NH2), 6.49 (d,
J = 8.2 Hz, 2H,ArH), 6.64 (d, J = 8.2 Hz, 2H,ArH).
Conclusion
An efficient approach to synthesising (E)-3-(aryl(arylamino)
methylene)indolin-2-one derivatives was developed and
12 target compounds were obtained in good yields. Their
antitumour activity in vitro was evaluated by the MTT assay,
and the results show that the introduction of a bromine atom
in the indolin-2-one and the piperidine-containing structure
appears to increase their antitumour activity. This could provide
a reference for new drug development.
Experimental
Synthesis of (E)-5-substituted-3-benzylideneindolin-2-one derivatives
(5a–d); general procedure
All reagents were purchased from commercial suppliers and were
used without further purification unless otherwise specified. Melting
points were determined by a capillary method and are uncorrected.
IR spectra were recorded on a Spectrum Two Li10014 spectrometer.
1H NMR and 13C NMR spectra were recorded on a Bruker Avance
400 MHz spectrometer. NMR spectra were recorded in DMSO-d6
or CDCl3 solutions at room temperature (20 2 °C). 1H and 13C
chemical shifts are quoted in parts per million (ppm) downfield from
tetramethylsilane (TMS). ESI-MS spectra were recorded on a Bruker
Esquire 3000 instrument. High-resolution mass spectra (HRMS)
were obtained on a MicroTOF-Q II mass spectrometer with an ESI
source (Waters, Manchester). All reactions were monitored by thin-
layer chromatography (TLC) and using ultraviolet light (254 nm). As
A mixture of compound 4 (10 mmol), benzaldehyde (12 mmol) and a
few drops of piperidine in ethanol (100 mL) was refluxed for 2 h. Part
of the solvent was evaporated under reduced pressure and the residue
was cooled to room temperature to crystallise the solid product,
which was filtered off, washed twice with ethanol, and dried to yield
compound 5.
(E)-3-Benzylideneindolin-2-one (5a): Yellow solid; yield 86%;
1
m.p. 177–178 °C (lit.24 178–179 °C); H NMR (400 MHz, DMSO-d6):
δ 6.87 (t, J = 7.6, 8.0 Hz, 1H, ArH), 6.92 (d, J = 7.6 Hz, 1H, ArH),
7.23–7.28 (m, 1H, ArH), 7.48–7.58 (m, 4H, ArH), 7.69 (s, 1H, =CH),
7.72 (d, J = 7.2 Hz, 2H, ArH), 10.69 (s, 1H, NH); 13C NMR (100 MHz,
DMSO-d
6): δ 169.06, 143.41, 136.17, 134.90, 130.56, 130.05, 129.66,
1
for the known compounds, melting points and H NMR spectra were
confirmed by comparison with data from the previously reported
129.14, 128.12, 122.75, 121.49, 121.32, 110.57; ESI-HRMS: Calcd for
C15H11NO: 244.0738 (M+Na+); found: 244.0729.

JOURNAL OF CHEMICAL RESEARCH 2018 47
(E)-3-Benzylidene-5-methylindolin-2-one (5b): Yellow solid; yield
82%; m.p. 196–198 °C; 1H NMR (400 MHz, DMSO-d6): δ 2.16 (s,
3H, CH3), 6.78 (d, J = 7.6, 1H, ArH), 7.04 (d, J = 7.6 Hz, 1H, ArH),
7.35 (s, 1H, ArH), 7.45–7.56 (m, 3H, ArH), 7.62 (s, 1H, =CH), 7.72
(d, J = 7.2 Hz, 2H, ArH), 10.52 (s, 1H, NH); 13C NMR (100 MHz,
DMSO-d6): δ 168.72, 143.34, 136.13, 134.68, 132.66, 130.43, 130.12,
129.58, 129.12, 128.06, 122.81, 121.35, 110.46, 20.82; ESI-HRMS:
Calcd for C16H13NO: 258.0895 (M+Na+); found: 258.0893.
(E)-3-Benzylidene-5-chloroindolin-2-one (5c): Yellow solid; yield
88%; m.p.227–229 °C; 1H NMR (400 MHz, DMSO-d6): δ 6.89 (d,
J = 8.4 Hz, 1H, ArH), 7.28 (dd, J = 8.4, 2.0 Hz, 1H, ArH), 7.41 (d,
J = 2.0 Hz, 1H, ArH), 7.52–7.56 (m, 3H, ArH), 7.71 (d, J = 7.2 Hz,
2H, ArH), 7.74 (s, 1H, =CH), 10.78 (s, 1H, NH); 13C NMR (100 MHz,
DMSO-d6): δ 168.66, 142.14, 138.13, 134.49, 130.51, 130.07, 129.62,
129.28, 127.36, 125.34, 122.93, 122.16, 111.93; ESI-HRMS: Calcd for
C15H10ClNO: 278.0349 (M+Na+); found: 278.0345.
2.0 Hz, 1H, ArH), 7.65 (d, J = 2.0 Hz, 1H, ArH), 11.01, 11.28 (double
singlet, 1H, NH); 13C NMR (100 MHz, DMSO-d6): δ 172.10, 141.22,
140.63, 135.16, 133.97, 130.99, 130.17, 129.89, 129.50, 128.42, 113.08,
57.77, 54.59; ESI-HRMS: Calcd for C15H10Br3NO: 481.8190 (79Br),
483.8169 (81Br) (M+Na+); found: 481.8186, 483.8163.
Synthesis of (E)-5-substituted-3-(phenyl(phenylamino)methylene)
indolin-2-one derivatives (8a–e) and (E)-5-substituted-3-(((4-
heterocyclic-2-oxoethoxy)phenylamino)(phenyl)methylene)indolin-
2-one derivatives (9a–g); general procedure
A mixture of compounds 6 (0.25 mmol) and 7 (or 3) (0.3 mmol) and
sodium bicarbonate (1.00 mmol) in DMF (50 mL) was reacted for
24 h at room temperature. The mixture was poured into water (30 mL),
the solid was filtered off, washed with water, and recrystallised from
methanol to yield compound 8 (or 9).
(E)-3-(Phenyl(phenylamino)methylene)-5-chloroindolin-2-one
(8a): Yellow solid; yield 68%; m.p. 217–218 °C; IR (KBr) (νmax/cm−1):
(E)-3-Benzylidene-5-bromoindolin-2-one (5d): Yellow solid;
1
3384 (–NH–C=O), 1644 (C=O), 1594 (–NH–); H NMR (400 MHz,
1
yield 85%; m.p. 213–214 °C (lit.25 212–213 °C); H NMR (400 MHz,
DMSO-d6): δ 6.57–6.72 (m, 3H, ArH), 6.93–7.05 (m, 2H, ArH),
7.07–7.27 (m, 5H, ArH), 7.36 (d, J = 8.7 Hz, 1H, ArH), 7.54 (dd, J = 8.8,
2.3 Hz, 1H, ArH), 7.72 (d, J = 2.3 Hz, 1H, ArH), 7.95 (s, 1H, NH),
11.84 (s, 1H, NH); 13C NMR (100 MHz, DMSO-d6): δ 162.31, 144.18,
144.03, 137.68, 134.77, 130.70, 130.57, 128.92, 127.73, 127.31, 125.68,
124.32, 122.67, 120.21, 118.96, 117.71, 117.56; ESI-HRMS: Calcd for
C21H16N2ClO: 347.0951 ([M+H]+); found: 347.0955.
DMSO-d6): δ 6.79 (dd, J = 14.4, 8.0 Hz, 1H, ArH), 7.40 (t, J = 10.0, 8.8
Hz, 1H, ArH), 7.49–7.59 (m, 4H, ArH), 7.69 (d, J = 6.4 Hz, 2H, ArH),
7.74 (s, 1H, =CH), 7.96 (d, J = 10.4 Hz, 1H, ArH), 10.79 (s, 1H, NH);
13C NMR (100 MHz, DMSO-d6): δ 168.58, 142.55, 138.14, 134.55,
130.56, 129.67, 129.30, 128.72, 127.31, 124.96, 123.48, 113.08, 112.48;
ESI-HRMS: Calcd for C15H10BrNO: 321.9843 (79Br), 323.9823 (81Br)
(M+Na+); found: 321.9840, 323.9833.
(E)-3-(4-Methylphenylamino(phenyl)methylene)-5-methylindolin-
2-one (8b): Yellow solid; yield 73%; m.p. 174–177 °C; IR (KBr)
Synthesis of 5-substituted-3-bromo-3-(bromo(phenyl)methyl)indolin-
2-one derivatives (6a–d); general procedure
1
(νmax/cm−1): 3364 (–NH–C=O), 1646 (C=O), 1585 (–NH–); H NMR
To a solution of compound 5 (5 mmol) in CH2Cl2 (100 mL), bromine
(10 mmol) was slowly added dropwise at 0–5 °C, and the solution was
then stirred for 3 h at room temperature. Water (50 mL) was poured
into the solution and the phase was separated. The organic layer was
washed with sodium thiosulfate solution to remove the extra bromine
and washed further with saturated salt water (30 mL) and dried
with anhydrous sodium sulfate. The organic layer was evaporated
under reduced pressure to give the crude product, which was then
recrystallised from ethanol to give compound 6.
3-Bromo-3-(bromo(phenyl)methyl)indolin-2-one (6a): Yellow
solid; yield 71%; m.p.193–194 °C (lit.26 194–195 °C); 1H NMR
(400 MHz, DMSO-d6): δ 5.96, 5.97 (double singlet, 1H, =CH), 6.86
(d, J = 8.3 Hz, 1H, ArH), 7.26 (d, J = 6.6 Hz, 3H, ArH), 7.33–7.39 (m,
2H, CH), 7.49 (dd, J = 8.4, 2.0 Hz, 2H, ArH), 8.10 (d, J = 2.0 Hz, H,
ArH), 11.01, 11.30 (double singlet, 1H, NH); 13C NMR (100 MHz,
DMSO-d6): δ 173.13, 141.23, 140.64, 135.15, 133.97, 130.99, 130.17,
129.89, 129.50, 128.28, 113.08, 57.78, 54.59; ESI-HRMS: Calcd for
C15H11Br2NO: 403.9085 (M+Na+); found: 403.9081.
3-Bromo-3- (bromo(phenyl)methyl)-5-methylindolin-2-one
(6b): Red solid; yield 76%; m.p.184–185 °C; 1H NMR (400 MHz,
DMSO-d6): δ 2.23 (s, 3H, CH3), 5.86, 5.97 (double singlet, 1H, =CH),
6.80–6.86 (m, 1H, ArH), 7.17 (s, 1H, ArH), 7.27 (d, J = 1.4 Hz, 1H,
ArH), 7.37–7.43 (dd, J = 5.1, 1.9 Hz, 3H, ArH), 7.49–7.59 (m, 3H,
ArH), 11.03, 11.30 (double singlet, 1H, NH); 13C NMR (100 MHz,
DMSO-d6): δ 172.32, 141.45, 140.35, 139.86, 135.22, 133.87, 130.94,
130.11, 129.48, 128.24, 115.03, 58.12, 54.58, 21.23; ESI-HRMS: Calcd
for C16H13Br2NO: 417.9241 (M+Na+); found: 417.9237.
(400 MHz, DMSO-d6): δ 2.25 (s, 6H, CH3), 6.55 (d, J = 8.0 Hz, 2H,
ArH), 6.86 (d, J = 8.0 Hz, 2H, ArH), 7.08–7.22 (m, 6H, ArH), 7.64 (d,
J = 1.5 Hz, 1H, ArH), 7.68 (d, J = 1.8 Hz, 1H, ArH), 7.70 (s, 1H, NH),
10.01 (s, 1H, NH); 13C NMR (100 MHz, DMSO-d6): δ 166.12, 162.11,
151.57, 145.63, 135.58, 134.78, 131.30, 130.35, 129.98, 128.73, 127.68,
127.20, 125.18, 122.28, 120.82, 119.27, 115.49, 20.58, 20.21; ESI-
HRMS: Calcd for C23H20N2O: 341.1654 ([M+H]+); found: 341.1660.
(E)-3-(4-Fluorophenylamino(phenyl)methylene)-5-methylindolin-
2-one (8c): Red solid; yield 70%; m.p. 196–198 °C; IR (KBr) (νmax
/
cm−1): 3368 (–NH–C=O), 1644 (C=O), 1588 (–NH–); 1H NMR
(400 MHz, DMSO-d6): δ 2.26 (s, 3H, CH3), 6.52–6.64 (m, 4H,
ArH), 7.08–7.22 (m, 6H, ArH), 7.63 (d, J = 1.5 Hz, 1H, ArH), 7.68 (d,
J = 1.6 Hz, 1H, ArH), 7.71 (s, 1H, NH), 9.96 (s, 1H, NH); 13C NMR
(100 MHz, DMSO-d6): δ 166.06, 162.09, 152.90, 145.66, 137.64,
134.84, 131.92, 130.89, 129.80, 129.25, 127.80, 127.15, 124.90, 120.41,
119.16, 119.01, 115.02, 20.72; ESI-HRMS: Calcd for C22H17FN2O:
345.1403 ([M+H]+); found: 345.1411.
(E) -5-Bromo-3- (4-methylphenylamino (phenyl) methylene)
indolin-2-one (8d): Yellow solid; yield 70%; m.p. >250 °C; IR (KBr)
1
(νmax/cm−1): 3360 (–NH–C=O), 1646 (C=O), 1590 (–NH–); H NMR
(400 MHz, DMSO-d6): δ 2.12 (s, 3H, CH3), 6.57 (d, J = 8.1 Hz, 2H,
ArH), 6.83 (d, J = 8.0 Hz, 2H, ArH), 7.08–7.26 (m, 5H, ArH), 7.30
(d, J = 8.7 Hz, 1H, ArH), 7.63 (dd, J = 8.7, 2.2 Hz, 1H, ArH), 7.76 (s,
1H, NH), 7.85 (d, J = 2.2 Hz, 1H, ArH), 11.81 (s, 1H, NH); 13C NMR
(100 MHz, DMSO-d6): δ 162.32, 144.35, 141.83, 138.00, 134.77,
133.14, 130.72, 129.42, 129.13, 127.78, 127.38, 127.27, 121.94, 119.31,
117.97, 113.37, 20.69; ESI-HRMS: Calcd for C22H17BrN2O: 405.0602
(79Br [M+H]+); found: 405.0600.
3-Bromo-3-(bromo(phenyl)methyl)-5-chloroindolin-2-one
(6c):
Yellow solid; yield 80%; m.p. 175–176 °C; 1H NMR (400 MHz,
DMSO-d6): δ 5.87, 5.95 (double singlet, 1H, =CH), 6.82 (d, J = 8.4 Hz,
1H, ArH), 7.22–7.29 (m, 3H, ArH), 7.35 (d, J = 4.9 Hz, 2H, ArH), 7.41
(dd, J = 8.3, 2.1 Hz, 1H, ArH), 7.98 (d, J = 2.1 Hz, 1H, ArH), 10.69,
11.02 (double singlet, 1H, NH); 13C NMR (100 MHz, DMSO-d6): δ
172.20, 141.51, 140.43, 139.84, 135.28, 133.91, 130.85, 130.18, 129.45,
128.43, 115.33, 58.12, 54.59; ESI-HRMS: Calcd for C15H10Br2ClNO:
437.8695 (M+Na+); found: 437.8703.
3,5-Dibromo-3-(bromo(phenyl)methyl)indolin-2-one (6d): Yellow
solid; yield 75%; m.p. 170–171 °C; 1H NMR (400 MHz, DMSO-d6): δ
5.86, 5.96 (double singlet, 1H, =CH), 6.86 (d, J = 8.3 Hz, 1H, ArH),
7.26 (d, J = 6.6 Hz, 3H, ArH), 7.33–7.39 (m, 2H, ArH), 7.49 (dd, J = 8.4,
(E)-5-Bromo-3-((4-fluorophenylamino)(phenyl)methylene)indolin-
2-one (8e): Green solid; yield 66%; m.p. >250 °C; IR (KBr) (νmax/cm−1):
3368 (–NH–C=O), 1648 (C=O), 1588 (–NH–); 1H NMR (400 MHz,
DMSO-d6): δ 6.55 (d, J = 8.0 Hz, 2H, ArH), 6.78 (d, J = 8.0 Hz, 2H, ArH),
7.07–7.25 (m, 5H, ArH), 7.32 (d, J = 8.5 Hz, 1H, ArH), 7.55 (dd, J = 8.8, 2.1
Hz, 1H, ArH), 7.76 (s, 1H, NH), 7.85 (d, J = 2.3 Hz, 1H, ArH), 11.82 (s, 1H,
NH); 13C NMR (100 MHz, DMSO-d6): δ 162.32, 144.30, 141.59, 138.15,
134.67, 133.24, 131.01, 129.32, 129.10, 127.71, 127.41, 127.19, 121.90,
119.28, 117.86, 111.27; ESI-HRMS: Calcd for C21H14BrFN2O: 409.0352,
411.0331 ([M+H]+); found: 409.0348, 411.0327.
(E)-3- (((4-Morpholino-2-oxoethoxy)phenylamino)(phenyl)
methylene)indolin-2-one (9a): Pink solid; yield 48%; m.p.

48 JOURNAL OF CHEMICAL RESEARCH 2018
225–226 °C; IR (KBr) (νmax/cm−1): 3342 (–NH–C=O), 1646 (C=O),
1588 (–NH–), 1232 (Ar–O), 1104 (CH2–O–CH2); 1H NMR (400 MHz,
DMSO-d6): δ 3.43 (t, J = 4.8 Hz, 4H, CH2), 3.56 (p, J = 4.6 Hz, 4H,
CH2), 4.64 (s, 2H, CH2), 6.61 (s, 4H, ArH), 7.07–7.20 (m, 6H, ArH),
7.28 (d, J = 8.7 Hz, 1H, ArH), 7.62 (dd, J = 8.7, 2.1 Hz, 1H, ArH), 7.71
(s, 1H, NH), 7.90 (d, J = 2.2 Hz, 1H, ArH), 11.71 (s, 1H, NH); 13C NMR
(101 MHz, DMSO-d6): δ 166.64, 162.36, 152.73, 144.68, 138.01,
137.61, 134.90, 133.07, 130.87, 127.74, 127.23, 127.13, 120.23, 119.70,
118.97, 117.93, 115.11, 113.28, 66.92, 66.52, 45.31; ESI-HRMS: Calcd
for C27H25N4O4: 456.1923 ([M+H]+); found: 456.1924.
(E)-5-Chloro-3-((4-(2-(4-morpholinyl)-2-oxoethoxy)phenylamino)
(phenyl)methylene)indolin-2-one (9b): Pink solid; yield 51%; m.p.
237–239 °C; IR (KBr) (νmax/cm−1): 3336 (–NH–C=O), 1648 (C=O),
1588 (–NH–), 1232 (Ar–O), 1108 (CH2–O–CH2); 1H NMR (400 MHz,
DMSO-d6): δ 3.43 (t, J = 4.8 Hz, 4H, CH2), 3.55 (p, J = 4.5 Hz, 4H,
CH2), 4.64 (s, 2H, CH2), 6.62 (s, 4H, ArH), 7.09–7.23 (m, 5H, ArH),
7.34 (d, J = 8.7 Hz, 1H, ArH), 7.52 (dd, J = 8.7, 2.3 Hz, 1H, ArH), 7.68
(s, 1H, NH), 7.74 (d, J = 2.3 Hz, 1H, ArH), 11.72 (s, 1H, NH); 13C NMR
(100 MHz, DMSO-d6): δ 166.65,162.42, 152.74, 144.74, 137.82, 137.72,
134.93, 130.85, 130.38, 127.76, 127.15, 125.45, 124.27, 120.19, 119.96,
118.48, 117.75, 115.15, 66.90, 66.52, 42.08; ESI-HRMS: Calcd for
C27H24ClN3O4: 490.1534 ([M+H]+); found: 490.1535.
(E)-5-Bromo-3-((4-(2-morpholinyl-2-oxoethoxy)phenylamino)
(phenyl)methylene)indolin-2-one (9c): Pink solid; yield 51%; m.p.
>250 °C; IR (KBr) (νmax/cm−1): 3338 (–NH–C=O), 1646 (C=O), 1588
(–NH–), 1232 (Ar–O), 1104 (CH2–O–CH2); 1H NMR (400 MHz,
DMSO-d6): δ 3.43 (t, J = 4.8 Hz, 4H, CH2), 3.55 (p, J = 4.5 Hz, 4H,
CH2), 4.64 (s, 2H, CH2), 6.61 (s, 4H, ArH), 7.08–7.21 (m, 5H, ArH),
7.28 (d, J = 8.7 Hz, 1H, ArH), 7.62 (dd, J = 8.7, 2.1 Hz, 1H, ArH), 7.70
(s, 1H, NH), 7.90 (d, J = 2.2 Hz, 1H, ArH), 11.71 (s, 1H, NH); 13C NMR
(100 MHz, DMSO-d6): δ 166.71, 162.44, 153.12, 144.84, 139.15,
137.84, 135.60, 132.40, 130.48, 127.84, 126.99, 125.51, 124.39, 120.20,
119.98, 117.93, 117.75, 115.36, 66.90, 66.52, 45.28; ESI-HRMS: Calcd
for C27H24BrN3O4: 534.1028 (79Br), 536.1008 (81Br [M+H]+); found:
534.1031, 536.1013.
137.60, 134.90, 133.07, 130.87, 127.75, 127.24, 127.14, 120.24, 119.73,
118.97, 117.94, 115.11, 113.28, 67.04, 46.13, 44.62, 41.61; ESI-HRMS:
Calcd for C28H28N4O3: 491.2059 ([M+Na]+); found: 491.2064.
(E)-5-Bromo-3-((4-(2-(4-methypiperazin-1-yl)-2-oxoethoxy)
phenylamino)(phenyl)methylene)indolin-2-one (9g): Grey solid;
yield 45%; m.p. 199–200 °C; IR (KBr) (νmax/cm−1): 3358 (–NH–
C=O), 1648 (C=O), 1592 (–NH–), 1232 (Ar–O); 1H NMR (400 MHz,
DMSO-d6): δ 2.18 (s, 3H, CH3), 2.27 (dt, J = 21.6, 4.9 Hz, 4H, CH2),
3.42 (d, J = 5.6 Hz, 4H, CH2), 4.61 (s, 2H, CH2), 6.61 (s, 4H, ArH),
7.08–7.21 (m, 5H, ArH), 7.28 (d, J = 8.8 Hz, 1H, ArH), 7.62 (dd, J = 8.7,
2.1 Hz, 1H, ArH), 7.70 (s, 1H, NH), 7.89 (d, J = 2.1 Hz, 1H, ArH),
11.71 (s, 1H, NH); 13C NMR (100 MHz, DMSO-d6): δ 166.39, 162.36,
152.80, 144.70, 138.02, 137.60, 133.07, 130.87, 127.75, 127.24, 127.14,
120.24, 119.73, 117.94, 115.11, 113.28, 67.04, 46.12, 44.62, 41.61; ESI-
HRMS: Calcd for C28H27BrN4O3: 547.1345, 549.1324 ([M+H]+); found:
547.1343, 549.1335.
Antitumour activity test
The antitumour activity of all of the synthesised compounds (8a–e
and 9a–g) against the cancer cell lines of HCT-116, A549, SKOV3
and MDA-MB-231 was measured by the MTT assay in the State
Key Laboratory of Biotherapy, Sichuan University. The results were
calculated as the mean values of three repeated experiments. Briefly,
cells (3 × 104 mL−1) were seeded in 96-well plates and cultured for 24 h,
followed by treatment with various concentrations of the compounds
ranging from 50 μM to 1.56 μM for 48 h at 37 °C under 5% CO2. Then,
MTT (20 μL of 5 mg mL−1) was added to each well and incubated for
another 4 h at 37 °C, the supernatant fluid was removed, and DMSO
(150 μL per well) was added for 15–20 min. Values of light absorption
(OD) were measured at 570 nm with a SpectraMAX M5 microplate
spectrophotometer (Molecular Devices). Gefitinib (TN) was used as
the reference drug for antitumour evaluation and was also assessed
under similar conditions for comparison with the tested compounds.
The response parameter calculated was the IC50 value, which
corresponds to the concentration required for 50% inhibition of cell
viability. The data for the antibacterial activity are presented in Table 1.
(E) -3- ((4- (2-Oxo-2- (piperidin-1-yl)ethoxy)phenylamino)
(phenyl)methylene)indolin-2-one (9d): Grey solid; yield 46%; m.p.
222–224 °C; IR (KBr) (νmax/cm−1): 3360 (–NH–C=O), 1640 (C=O),
1592 (–NH–), 1234 (Ar–O); 1H NMR (400 MHz, DMSO-d6): δ
1.38–1.61 (m, 6H, CH2), 4.35 (t, J = 5.1 Hz, 4H, CH2), 4.59 (s, 2H,
CH2), 6.61 (s, 4H, ArH), 7.07–7.22 (m, 6H, ArH), 7.28 (d, J = 8.7 Hz,
1H, ArH), 7.62 (dd, J = 8.7, 2.2 Hz, 1H, ArH), 7.70 (s, 1H, NH), 7.90
(d, J = 2.2 Hz, 1H, ArH), 11.70 (s, 1H, NH); 13C NMR (100 MHz,
DMSO-d6): δ 166.05, 162.36, 152.89, 144.71, 138.01, 137.52, 134.90,
133.06, 130.88, 127.73, 127.23, 127.11, 120.30, 119.61, 118.96, 117.93,
115.05, 113.28, 67.27, 42.63, 26.42, 25.76; ESI-HRMS: Calcd for
C28H27N3O3: 476.1950 ([M+Na]+); found: 476.1948.
Acknowledgements
We thank the Analytical and Testing Center of West China
Medical College, Sichuan University for 1H NMR and IR
measurements and the State Key Laboratory of Biotherapy,
West China Hospital, Sichuan University for 13C NMR and
HRMS measurements.
Electronic Supplementary Information
The ESI is available through: http://ingentaconnect.com/
content/stl/jcr/2018/00000042/00000001/art00011
(E)-5-Bromo-3-((4-(2-oxo-2-(piperidin-1-yl)ethoxy)phenylamino)
(phenyl)methylene)indolin-2-one (9e): Red solid; yield 50%; m.p.
204–206 °C; IR (KBr) (νmax/cm−1): 3366 (–NH–C=O), 1644 (C=O),
1592 (–NH–), 1232 (Ar–O); 1H NMR (400 MHz, DMSO-d6): δ
1.38–1.61 (m, 6H, CH2), 4.35 (t, J = 5.1 Hz, 4H, CH2), 4.59 (s, 2H,
CH2), 6.62 (s, 4H, ArH), 7.06–7.23 (m, 5H, ArH), 7.29 (d, J = 8.7 Hz,
1H, ArH), 7.60 (dd, J = 8.7, 2.1 Hz, 1H, ArH), 7.70 (s, 1H, NH), 7.90
(d, J = 2.3 Hz, 1H, ArH), 11.70 (s, 1H, NH); 13C NMR (100 MHz,
DMSO-d6): δ 166.14, 163.43, 154.49, 146.61, 137.67, 137.19, 133.08,
132.87, 130.21, 129.12, 128.67, 128.36, 122.23, 120.41, 117.91, 117.33,
115.60, 114.11, 68.09, 43.27, 26.51, 25.59; ESI-HRMS: Calcd for
C28H26BrN3O3: 534.1215 (81Br [M+H]+); found: 534.1213.
Received 31 October 2017; accepted 5 January 2018
Paper 1705076
https://doi.org/10.3184/174751918X15166485941737
Published online: 28 January 2018
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