Synthesis and anti-cancer activity evaluation of 5-(2-carboxyethenyl)-isatin derivatives

Article abstract of DOI:10.1016/j.ejmech.2015.12.050

A series of novel di- or trisubstituted isatin derivatives were designed and synthesized in 5–6 steps in 25–45% overall yields. Their structures were confirmed by 1H NMR and 13C NMR as well as LC-MS. The anticancer activity of the fourty-three new isatin derivatives against human T lymphocyte cells Jurkat was evaluated by MTT assay in vitro. SAR study suggested that the combination of 1-benzyl and 5-[trans-2-(methoxycarbonyl)ethen-1-yl] substitution greatly enhanced their cytotoxic activity. Among them, compound 2h was shown to have a significant cytotoxic activity with an IC50 value of 0.03 μM, more than 330-fold higher than that of it's mother molecule isatin. Investigation of the cell morphology changes and annexin-V/PI staining study demonstrated that compound 2h inhibited the proliferation of Jurkat cells by inducing apoptosis. Since compound 2h induced the dissipation of mitochondrial membrane potential and the activation of caspase-3, it was obvious that compound 2h inhibited the proliferation of Jurkat cells through the mitochondrial apoptotic pathway. Other than this, compound 2h exerted inhibition effect to many other tumor cells and only showed weak cytotoxic to human normal cells suggesting that compound 2h possessed a broad range of anticancer spectrum and high safety to normal cells.

Full text of DOI:10.1016/j.ejmech.2015.12.050

Accepted Manuscript
Synthesis and Anti-cancer Activity Evaluation of 5-(2-Carboxyethenyl)-isatin
Derivatives
Teng Yu-Ou, Zhao Hong-Ye, Wang Jing, Liu Huan, Gao Mei-Le, Zhou Yao, Han Kai-
Lin, Fan Zhen-Chuan, Zhang Yong-Min, Sun Hua, Yu Peng
PII:
S0223-5234(15)30436-0
10.1016/j.ejmech.2015.12.050
EJMECH 8290
DOI:
Reference:
To appear in: European Journal of Medicinal Chemistry
Received Date: 2 December 2015
Revised Date: 30 December 2015
Accepted Date: 31 December 2015
Please cite this article as: T. Yu-Ou, Z. Hong-Ye, W. Jing, L. Huan, G. Mei-Le, Z. Yao, H. Kai-Lin, F.
Zhen-Chuan, Z. Yong-Min, S. Hua, Y. Peng, Synthesis and Anti-cancer Activity Evaluation of 5-(2-
Carboxyethenyl)-isatin Derivatives, European Journal of Medicinal Chemistry (2016), doi: 10.1016/
j.ejmech.2015.12.050.
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ACCEPTED MANUSCRIPT
Graphical Abstract
A series of di-or trisubstituted novel isatin derivatives were synthesized. The anti-cancer activity
evaluation results revealed these isatin derivatives inhibit Jurkat proliferation by inducing
mitochondrial-mediated apoptosis.
2
R = H, F, Cl, Br, CH
3
,
O
X,Y: =O; H, H; H, OH; =N-OH;
O
OCH , But,
3
O
OR
O
X
Y
OPh
Me
Et
Br
R2
O
NH2
O
N
Cl
Cl
R1
N
Br
O
O
N
X
(
X=Br, CF , CN, H, CH , F, Cl)
3 3

ACCEPTED MANUSCRIPT
Synthesis and Anti-cancer Activity Evaluation of
5
-(2-Carboxyethenyl)-isatin Derivatives
a
a
a
a
a
Yu-Ou Teng , Hong-Ye Zhao , Jing Wang , Huan Liu , Mei-Le Gao , Yao
a
a
b
c
a
a*
Zhou , Kai-Lin Han , Zhen-Chuan Fan , Yong-Min Zhang , Hua Sun , Peng Yu
a
Key Lab of Industrial Fermentation Microbiology,Tianjin Key Lab of
Industrial Microbiology, Sino-French Joint Lab of Food Nutrition/Safety and
Medicinal Chemistry, Tianjin University of Science and Technology, Tianjin,
3
00457, P.R.China
b
Key Laboratory of Food Nutrition and Safety (Tianjin University of Science
and Technology), Ministry of Education, Tianjin 300457, PR China
c
Université Pierre et Marie Curie-Paris 6, Institut Parisien de Chimie
Moléculaire UMR CNRS 8232, 4 place Jussieu, 75005, Paris, France
*
Corresponding author. Tel.: +86 22 60912562, E-mail: yupeng@tust.edu.cn
1

ACCEPTED MANUSCRIPT
Abstract: A series of novel di- or trisubstituted isatin derivatives were
designed and synthesized in 5-6 steps in 25-45% overall yields. Their
1
13
structures were confirmed by H NMR and C NMR as well as LC-MS.
The anticancer activity of the fouty-three new isatin derivatives against
human T lymphocyte cells Jurkat was evaluated by MTT assay in vitro.
SAR study suggested that the combination of 1-benzyl and
5
-[trans-2-(methoxycarbonyl)ethen-1-yl] substitution greatly enhanced
their cytotoxic activity. Among them, compound 2h was shown to have a
significant cytotoxic activity with an IC value of 0.03 ꢀM, more than
5
0
3
30-fold higher than that of it’s mother molecule isatin. Investigation of
the cell morphology changes and annexin-V/PI staining study
demonstrated that compound 2h inhibited the proliferation of Jurkat cells
by inducing apoptosis. Sicne compound 2h induced the dissipation of
mitochondrial membrane potential and the activation of caspase-3, it was
obvious that compound 2h inhibited the proliferation of Jurkat cells
through the mitochondrial apoptotic pathway. Other than this, compound
2
h exerted inhibition effect to many other tumor cells and only showed
weak cytotoxic to human normal cells suggesting that compound 2h
possessed a broad range of anticancer spectrum and high safety to normal
cells.
Keywords: Isatin derivatives; Anti-tumor agents; Jurkat cells;
Proliferation; Apoptosis
2

ACCEPTED MANUSCRIPT
1
. Introduction
Isatin as an indole derivative widely presents endogenously in both
human and other mammalian tissues and fluids likely as a result of the
tryptophan metabolic pathway. The versatility molecular architecture
isatin makes it as an ideal platform for structural modification and
derivatization and many isatin derivatives exhibit a broad range of
biological activities such as anticancer[1], antidepressant[2],
anticonvulsant[3], antifungal[4], anti-HIV[5] and anti-inflammatory
activity [6]. In the last several decades, many researchers have embarked
on the development of new isatin-based anticancer agents[7-9]. Eshba’s
group found that 5-bromoisatin 3-(2-nitrophenyl) hydrazone and a series
of 5-[5-bromo-(2-oxo-3-indolinylidene)]amino thiazolidine-2,4-diones
substituted by various Mannich bases exhibit antileukemic activity
against P38 lymphocytic leukemia in mice [10]. Popphas reported that
some 3-hydroxy-3-substituted oxindoles obtained from condensation of
substituted isatins with cyclic ketones possess anticonvulsant activity[11].
SU11248 (Sutent), a 5-fluoro-3-substituted isatin derivative, was
approved by the FDA in 2006 for treatment of advanced renal carcinoma
and gastrointestinal stromal tumors [12, 13]. C5- and C6-substituted
isatin analogues were selective about MAOB inhibitors with
5
1
-(4-phenylbutyl)isatin exhibiting the highest activity and being
8,500-fold more potent than isatin[14]. Recently, the groups of Mach[15,
3

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1
6] and Reichert[17] synthesized several N alkyl and C-5 sulfonamido
isatin analogues as small molecule caspase-3 and caspase-7 inhibitors.
Many cytotoxic agents and/or DNA damaging agents could inhibit
the cancer cell proliferation by cell cycle arrest. The cell cycle is
mediated by the activation of cyclin-dependent kinases (Cdks), which is
activated by forming complexes with cyclins [18]. The cyclin B1/Cdk1
complex is the primary regulator of transition from the G to M phase.
2
Apoptosis is the result of a highly complicated cascade of cellular events
that result in cell rounding and shrinkage, chromatin condensation, DNA
fragmentation, shedding of smaller fragments from cells, and detachment
from the plate[19]. In particular, the mitochondria plays a central role in
the occurrence of apoptosis resulting from many chemotherapeutic
agents[20]. It is also well known that caspases, such as caspase-3, -8 and
-9, which are highly conserved cysteine-dependent aspartate-specific
proteases and the key components of the apoptotic machinery, dominate
in apoptotic progression[21, 22].
As part of our research program on the SAR study of isatin
derivatives’ anticancer property and based on our previous short
communication[23, 24], herein we wish to report in details the synthesis
and anticancer activity evaluation of a series of N-benzylated isatins
possessing an acrylate moiety at C-5 against human leukemia Jurkat cells
and the potential molecular mechanism of the cytotoxic activity.
4

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2
. Chemistry
As shown in Scheme 1, 5-substituted isatins 1a-1g were prepared by two
steps in 55-80% yield by literature procedures [25]. Microwave-assisted
Heck coupling reaction was employed to converted 5-bromo isatin (1d)
5
-trans-(2-methoxycarbonylethenyl) isatin (1h) in 70%. Further
derivatization led to the synthesis of compounds 2a-2p in good to
excellent yield by N-alkylation with alkyl halides in the presence of
K CO . Target compounds 2q-2o were synthesized by
2
3
microwave-assisted Heck coupling reaction in a similar way for the
synthesis of 2h. Compound 3p-3w were synthesized from 2h and 2k
through an appropriate carbonyl group derivatization reaction (Scheme
2
).
3
. Results and discussion
Based on the identification of several major structure motifs that main
potentially improve isatin’s antitumor activity as illustrated in Figure 1,
the investigation was started with phenyl portion’s substitution pattern
which was followed by N-derivatization and then C-3 variation.
The in vitro anti-tumor activity of the newly synthesized isatin
derivatives on human tumor cell Jurkat was evaluated by MTT assay with
Camptothecin (CPT) as the positive control [26].
5

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As shown in Table 1, isatin derivatives 1a-1f which possess simple
substituent groups on the C-5 position with different sizes and varing
electronic properties have slight cytotoxicty effect. Interestingly, when a
methyl acrylate group was introduced at C-5, compound 1h showed at
least 200 times stronger than isatin itself. However, compound 1g, which
contains an n-butyl group at C-5, had no improvement. This observation
suggests that the good inhibitory activity of compound 1h does not
simply correlate with the size of the acrylate moiety and the limited
spatial orientation due to its conjugation with the aromatic ring may play
a role.
With the identification of compound 1h as a potential lead, the SAR
study then focused on the N-derivatized analogues. Recently, some
N-benzyl isatin derivatives were reported to exhibit a broad range of
biological activities such as analgestic effect [27] and cytotoxicity[28]
and they also could be used as asreverse transcriptase inhibitors[15, 16,
2
9] and caspase inhibitors [30] . Therefore, it seemed strongly possible
that the combination of C-5 olefination with an acrylate moiety and
N-substitution may further enhance their potency.
As shown in Table 2, simple alkyl groups such as the N-methyl and
N-ethyl (compounds 2a and 2b) did not give a significant change in the
cell growth inhibition compared to compound 1h. This observation
suggested that the N-H group is not a necessary structure unit for the
6

ACCEPTED MANUSCRIPT
activity of compound 1h. Since the N-methyl and N-ethyl groups maybe
too small to cause a significant effect to compound 1h potency, a benzyl
group was then introduced to the N-1 position to obtain compound 2c.
Fortunately, compound 2c was found to have a 3 fold improvement in its
potency, suggesting that larger groups at the N-1 position could increase
the potency. Further test on compounds 2d-2f (with an electron
withdrawing group) and 2g, 2p (with an electron donating group) showed
that these derivatives had no different effect to the benzyl 2c.
Compounds 2h and 2k containing a 4-methoxy and 4-bromo benzyl
group, respectively, exhibited excellent cytotoxic. For Jurkat, the IC of
5
0
compounds 2h and 2k reached 0.03 ꢀΜ and 0.06 ꢀΜ, respectively. It was
also noticed that the potency of the meta and ortho bromo-substituted
benzyl compounds 2l (0.12 ꢀΜ) and 2m (0.80 ꢀΜ) dropped by 4-27 folds
as compared to their para counterpart 2h (0.03 ꢀΜ). Moreover, when the
N-1 substitution was changed to the more bulky 2-naphthylmethyl (2n) or
the phenoxyethyl (2o) groups the potency also diminished. The results
above illustrated the importance of the size and orientation for
N-substitution in the acrylate-containing isatin analogues for their
anticancer activity.
To clarify the importance of the position of the acrylate moiety in
2
h, regioisomers 2q-2s were synthesized by using a similar procedure for
the synthesis of 2h (Table 3). The 6- and 7-substituted 2r and 2s were
7

ACCEPTED MANUSCRIPT
found to be much less potent than 2h (100 folds) and the 4-substituted
counterpart 2q (26 folds). Along with the inferior potency of 2q as
compared to 2h, these data suggest that the presence of a methyl acrylate
group at the para-position found in 2h is vital to maintain a high level of
potency, probably as the result of electronic and/or geometric preference.
After the N-substitution and C-substitution patterns were
established, target compounds 3a-3o were synthesized by
microwave-assisted Heck coupling reaction in a similar way for the
synthesis of compound 2h and tested to dertermine their antitumor
activity (Table 4). The results indicated that the potency generally
decreased with the increase in the size of the alkyl group (3a, 3c, 3g and
3
e vs. 2h; 3b, 3d, 3h and 3f vs. Compound 2h in going from Me to Et,
Bu, 2-hydroxyethyl and t-Bu). The presence of a free acrylic acid (3i)
brought about a drastic diminishment in potency, pointing to a
detrimental effect of the highly polar and readily ionizable free carboxyl
group. All the amide analogues examined (3j to 3o) were found to
possess low activities relative to the esters either duo to the high polarity
of these groups or an increase in their sized in the case of 3l/3m and
3
n/3o.
To examine the importance of the C-3 carbonyl functionality, eight
new isatin derivatives (3p-3w) were synthesized from compounds 2h and
2
k through an appropriate carbonyl group derivatization reaction
8

ACCEPTED MANUSCRIPT
(Scheme 2). All these derivatives proved to be inferior candidates as
inhibitors of Jurkat (Table 5), suggesting that the carbonyl functionality at
C-3 is essential to maintain the observed high antitumor activity.
The SAR study of these compounds led to the identification of the
nevel isatin derivative, compound 2h, as a highly potent anticancer
compound with an IC of 0.03 ꢀΜ against human leukemia Jurkat cells.
5
0
After then, the molecular mechanism by which compound 2h exerts its
cytotoxic activity in Jurkat cells was investigated.
It was obvious that treatment with compound 2h caused a typical
apoptotic morphology change in Jurkat cells, such as cell shrinkage
and/or blebbing. As shown in Figure 2A, the apoptotic cells were clearly
observed in Jurkat cells after treatment with compound 2h for 12 h. Flow
cytometic analysis also showed that apoptotic cells appeared with a
time-dependent pattern in compound 2h-treated Jurkat cells when the
cells were double labelled with annexin-V-FITC and PI (Figure 2B). After
+
treatment with compound 2h, the apoptotic rates (Annexin V ) of Jurkat
cells were increased to 42.0% of the total cells (0.03 ꢀΜ, 12 h) whereas
only few cells were observed as apoptotic cells in the control.
Mitochondrial pathway is an important pathway for
chemotherapeutic agents to induce cell apoptosis. After treatment with
compound 2h, the dissipation of mitochondrial membrane potential was
clearly observed in Jurkat cells. The dissipation rates of mitochondrial
9

ACCEPTED MANUSCRIPT
membrane potential of 2h-treated Jurkat cells were increased to 81.9% of
the total cells (48 h) whereas only 7.8% cells were observed as apoptotic
cells in the control. This result indicated the possible role of
mitochondrial pathway in 2h-caused apoptosis. This conlcusion was
further supported by the finding that the caspase-3 was activated in
2
h-treated Jurkat cells. Caspase family proteins play an important role in
metochandrial-mediated apoptosis. The activation of caspase-3 plays a
central role in the initiation of apoptosis. In the present study, the
activation of caspase-3 was quantified by means of a fluorescence-based
method. The result in Figure 3B showed that compound 2h caused the
activation of caspase-3, which peaked at 48 h following treatment. Based
on the above observations, it was suggested that compound 2h induced
mitochondrial dependent apoptosis in Jurkat cells.
Our preliminary experimental results showed that the compound 2h
had obvious inhibitory effect to the Jurkat cells. To further investigate the
anticancer spectrum of compound 2h, a comparative study was carried
out on thirteen human cancer cell lines including Jurkat, BT549, BT474,
MCF-7, HT-29, HCT-116, HeLa, HepG2, A498, NCI-H1299, A549,
DU-145 and SK-OV-3 as well as two normal human cell lines HUVEC
and HEK293. As shown in Table 6, the result showed that the treatment
of compound 2h efficiently inhibited the proliferation of most tumor cells,
suggesting that compound 2h has a superior anticancer action against
1
0

ACCEPTED MANUSCRIPT
various cancers and exhibit a broader anticancer spectrum against various
cancer cell lines. It is noteworthy that, as compared to the cytotoxic
potency against tumor cell lines, compound 2h possessed low
cytotoxicity to HUVEC and 293 cells (IC >10 ꢀΜ).
50
Taken together, these results suggested that compound 2h possesses
low cytotoxicity to normal human cells and potential anti-cancer activity
against human Jurkat cells by enhancing mitochondrial-mediated
apoptosis.
4
. Conclusions
In summary, forty three di- or trisubstituted novel isatin derivatives
were designed and synthesized in 5-6 steps in 25-45% overall yields.
The anticancer activity of the 43 newly synthesized isatin derivatives
against human tumor cell line Jurkat was evaluated and the test illustrated
that the combination of 1-benzyl and
5
-[trans-2-(methoxycarbonyl)ethen-1-yl] substitution greatly enhanced
their cytotoxic activity whereas an intact carbonyl functionality on C-3 as
present in the parent ring is required to such a potency.
1
-(4-methoxyl)benzyl 5-(2-carboxyethenyl) isatin derivative (named
2
h) was selected as a highly potent anticancer compound with an IC50
vaalue of as low as 0.03 ꢀΜagainst human leukemia Jurkat cells. The
cytotoxic activity of 2h on Jurkat cells lie in the inhibition of the
1
1

ACCEPTED MANUSCRIPT
proliferation of Jurkat cells by inducing mitochondrial-mediated
apoptosis. Further in vivo test of compound 2h is ongoing in our lab.
5
5
. Experimental section
.1. Chemistry
All chemicals (reagent grade) used were purchased from Sigma-Aldrich
U.S.A) and Aladdin-Reagent Co., Ltd (China). The solvents for reaction
were distilled over Na (for toluene and THF) or CaH (for CH Cl ) under
(
2
2
2
a nitrogen atmosphere. All reactions were carried out in oven-dried
glassware under an inert atmosphere (nitrogen or argon). Microwave
reaction was carried out in Biotage Initiator made in Sweden.TLC was
run on the silica gel coated aluminum sheets (Silica Gel 60GF254, E.
1
13
Merck, Germany). H NMR and C NMR were recorded on a Bruker
AVANCE instrument (400 MHz).ESI mass spectra were obtained on an
LCQ-Advantayc-MAX (LAM10188, Finnigan, Co., Ltd, USA). The
purity of the final products was determined by HPLC (LabTech) on a
Diamonsil C column (4.6 mm×250 mm, 5µm) with methanol/H O
18
2
(
90/10 v/v) at 0.5 ml/min flow rate and 254 nm detector wavelength.
.1.1 Synthesis of compounds 1a-1h and 2a-2p
5
The general procedure for the preparation of isatin derivatives was as
follows. A mixture of 0.05 mol substituted anilines, 0.15 mol
Hydroxylammonium chloride, 0.35 mol Na SO , 2 mol/L Hydrochloric
2
4
1
2

ACCEPTED MANUSCRIPT
acid at 5 ml and Chloral hydrate at 0.06 mol were stirred in 250 ml H O
2
at 95 ℃. The progress of the reaction was monitored by TLC (petroleum
ether/ethyl acetate). After the reaction finished, the reaction mixture was
filtered, dryness. The crude product was used directly for the next step
without further purification.
To a flask (100 ml) which contained concentrated sulfuric acid (20
ml) was added N-2-(hydroxyimino) acetamide derivatives (7.0 g) in
portions at 50 ℃with vigorous stirring. The reaction temperature was
maintained at 50 ℃-75 ℃during the addition. After the addition was
completed, the mixture was heated to 80 ℃and stir at 80 ℃for 30 min.
The reaction mixture was cooled to room temperature and then poured
onto ice (250 g). The solid which resulted was filtered out and dried over
air to yield the crude which was purified by dissolving in dilute sodium
hydroxide (5%, 100 ml) followed by acidified with 4N hydrochloric acid
(20 ml). The solid which formed was filtered out and dried over air to
provide the purified compounds 1a-1g.
To a microwave reactor vial (5 ml) which contained the solution of
5
-bromoindoline-2,3-dione (1.0 g, 4.4 mmol) in DMF (5 ml) were added
PdCl (PPh3) (155 mg, 0.22 mmol), CH COOK (0.6 g, 6.2 mmol) and
2
2
3
Methyl acrylate (0.45 g, 5.3 mmol) under the atmosphere of argon. The
microwave reactor vial was caped and placed into the microwave cavity.
The reaction mixture was irradiated at high level for 30 min at 145 ℃.
1
3

ACCEPTED MANUSCRIPT
The reaction mixture was cooled to room temperature, poured onto 100
ml ice-water and then extracted with dichloromethane (3×100 ml). The
combined organic layers were washed with water (2×100 ml), brine (100
ml) and dried over anhydrous magnesium sulfate. The solvent was
removed under vacuum to afford the crude which was purified by flash
column chromatography (silica gel, petroleum ether/ethyl acetate 3:1) to
yield the desired compound 1h.
To a flask (25 ml) which contained the solution of compound 1h
(0.2 g, 0.86 mmol) in dry N,N-dimethylformamide (2 ml). The reaction
temperature was maintained at 0 ℃ followed by addition K CO (0.35 g,
2
3
2
.6 mmol). Stirring 5 min, was added alkyl or benzyl (1.0 mmol) and the
mixture allowed warm to room temperature. The reaction mixture was
stirred at room temperature for 4 h. The orange solution was poured water
(25 ml) and extracted with dichloromethane (3 × 100 ml). The combined
extracts were dried over anhydrous magnesium sulfate, filtered and
evaporated. Chromatography (petroleum ether/ethyl acetate 10:1)
afforded the title compounds 2a-2p.
5
.1.1.1. (E)-methyl 3-(2,3-dioxoindolin-5-yl)acrylate (1h)
1
Yield 70%, purity 98%; H NMR (400 MHz, DMSO-d ) δ 3.71(s, 3H),
6
6
1
.60-6.64(d, 1H, J = 16.0 Hz), 6.92-6.94(d, 1H, J = 8.0 Hz), 7.63-7.67(d,
13
H, J = 16.0 Hz), 7.91-7.95(m, 2H), 11.25(s, 1H). C NMR (100 MHz,
DMSO-d6) δ 51.89, 112.92, 117.24, 118.89, 124.94, 129.35, 138.59,
1
4

ACCEPTED MANUSCRIPT
43.89, 152.36, 160.03, 167.17, 184.33. ESI-MS: 232.1 [M+H]+,
54.1[M+Na]+.
1
2
5
.1.1.2. (E)-methyl 3-(1-methyl-2,3-dioxoindolin-5-yl)acrylate (2a)
1
Yield 45%, purity 98%; H NMR (400 MHz, CDCl ) δ 3.29(s, 3H),
3
3
7
.81(s, 3H), 6.37-6.41(d, 1H, J = 16.0 Hz), 6.93-6.95(d, 1H, J = 8.4 Hz),
13
.60-7.64(d, 1H, J = 16.0 Hz), 7.74-7.76(m, 1H), 7.78(s, 1H). C NMR
(
100 MHz, CDCl ) δ 26.46, 51.87, 110.37, 117.84, 118.16, 124.01,
3
1
30.55, 138.17, 142.47, 152.24, 158.13, 166.94, 182.71. ESI-MS: 246.1
[M+H]+.
5
.1.1.3. (E)-methyl 3-(1-ethyl-2,3-dioxoindolin-5-yl)acrylate (2b)
1
Yield 43%, purity 100%; H NMR (400 MHz, CDCl ) δ 1.31-1.35(t, 3H),
3
3
.81(s, 3H), 3.82(m, 2H), 6.37-6.41(d, 1H, J = 16.0 Hz), 6.94-6.96(d, 1H,
J = 8.0 Hz), 7.61-7.65(d, 1H, J = 16.0 Hz), 7.72-7.74(m, 1H), 7.78 (s,
1
3
1
H). C NMR (100 MHz, CDCl ) δ 12.56, 35.25, 51.85, 110.40, 118.03,
3
118.10, 124.22, 130.36, 138.06, 142.47, 151.50, 157.78, 166.95, 183.04.
ESI-MS: 260.1 [M+H]+, 282.1[M+Na]+.
5
.1.1.4. (E)-methyl 3-(1-benzyl-2,3-dioxoindolin-5-yl)acrylate (2c)
1
Yield 44%, purity 98%; H NMR (400 MHz, CDCl ) δ 3.79 (s, 3H),
3
4
7
.95(s, 2H), 6.33-6.37(d, 1H, J = 16.0 Hz), 6.80-6.82(d, 1H, J = 8.0 Hz),
1
3
.29-7.38(m, 5H), 7.58(d, 1H, J = 16.0 Hz), 7.61(m, 1H), 7.78(s, 1H). C
NMR (100 MHz, CDCl ) δ 44.28, 51.87, 111.42, 118.08, 118.23, 124.15,
3
1
5

ACCEPTED MANUSCRIPT
27.45, 127.45, 128.37, 129.17, 129.17, 130.59, 134.15, 137.98, 142.40,
51.50, 158.14, 166.91, 182.64. ESI-MS: 322.2 [M+H]+.
.1.1.5. (E)-methyl
1
1
5
3
-(1-(4-fluorobenzyl)-2,3-dioxoindolin-5-yl)acrylate (2d)
1
Yield 43%, purity 99%; H NMR (400 MHz, CDCl ) δ 3.80(s, 3H),
3
4
7
7
5
1
.92(s, 2H), 6.34-6.38(d, 1H, J = 16.0 Hz), 6.80-6.82(d, 1H, J = 8.4 Hz),
.03-7.07(m, 2H), 7.30-7.33(m, 2H), 7.57-7.61(d, 1H, J = 16.0 Hz),
1
3
.62-7.65(m, 1H), 7.79(s, 1H). C NMR (100 MHz, CDCl ) δ 43.59,
3
1.87, 111.24, 116.06, 116.27, 118.09, 118.35, 124.23, 129.27, 129.35,
29.98, 130.02, 130.71, 137.99, 142.31, 151.23, 158.10, 166.88, 182.48.
ESI-MS: 340.1 [M+H]+, 362.1 [M+Na]+.
5
3
.1.1.6. (E)-methyl
-(1-(4-cyanobenzyl)-2,3-dioxoindolin-5-yl)acrylate (2e)
1
Yield 40%, purity 98%; H NMR (400 MHz, CDCl ) δ 3.80(s, 3H),
3
5
7
3
.00(s, 2H), 6.35-6.39(d, 1H, J = 16.0 Hz), 6.74-6.76(d, 1H, J = 8.4 Hz),
.44-7.46(d, 2H, J = 8.4 Hz), 7.57-7.61(d, 1H, J = 16.0 Hz), 7.64-7.68(m,
13
H), 7.82(s, 1H). C NMR (100 MHz, CDCl ) δ 43.83, 51.92, 110.93,
3
112.58, 118.11, 118.14, 118.70, 124.45, 128.04, 128.04, 131.11, 132.99,
1
3
5
3
32.99, 138.06, 139.51, 142.09, 150.72, 158.08, 166.79, 181.94. ESI-MS:
47.2 [M+H]+.
.1.1.7. (E)-methyl
-(2,3-dioxo-1-(4-(trifluoromethyl)benzyl)indolin-5-yl)acrylate (2f)
1
6

ACCEPTED MANUSCRIPT
1
Yield 44%, purity 98%; H NMR (400 MHz, CDCl ) δ 3.80(s, 3H),
3
5
7
7
.01(s, 2H), 6.35-6.39(d, 1H, J = 16.0 Hz), 6.76-6.78(d, 1H, J = 8.4 Hz),
.44-7.46(d, 2H, J = 8.0 Hz), 7.59(d, 1H, J = 16.0 Hz), 7.63(m, 3H),
1
3
.81(s, 1H). C NMR (100 MHz, CDCl ) δ 43.79, 51.90, 111.11, 118.11,
3
1
18.53, 124.36, 126.14, 126.18, 126.22, 126.26, 127.72, 127.72, 130.94,
38.07, 138.23, 142.20, 150.96, 158.11, 166.84, 182.18. ESI-MS: 390.1
M+H]+, 412.1 [M+Na]+.
1
[
5
3
.1.1.8. (E)-methyl
-(1-(4-methylbenzyl)-2,3-dioxoindolin-5-yl)acrylate (2g)
1
Yield 42%, purity 98%; H NMR (400 MHz, CDCl ) δ 2.32(s, 3H),
3
3
.79(s, 3H), 4.90(s, 2H), 6.32-6.36(d, 1H, J = 16.0 Hz), 6.81-6.83(d, 1H,
J = 8.4 Hz), 7.14-7.16(d, 2H, J = 8.0 Hz), 7.20-7.22(d, 2H, J = 8.0 Hz),
1
3
7
.58(d, 1H, J = 16.0 Hz), 7.61(m, 1H), 7.77(s, 1H). C NMR (100 MHz,
CDCl ) δ 21.12, 44.05, 51.86, 111.48, 118.06, 118.13, 124.10, 127.48,
3
1
1
5
3
27.48, 129.80, 129.80, 130.48, 131.09, 137.98, 138.21, 142.45, 151.58,
58.14, 166.94, 182.75. ESI-MS: 336.2 [M+H]+, 358.1 [M+Na]+.
.1.1.9. (E)-methyl
-(1-(4-methoxybenzyl)-2,3-dioxoindolin-5-yl)acrylate (2h)
1
Yield 46%, purity 99%; H NMR (400 MHz, CDCl ) δ 3.78(s, 3H),
3
3
.80(s, 3H), 4.88(s, 2H), 6.33-6.37(d, 1H, J = 16.0 Hz), 6.83-6.85(d, 1H,
J = 8.4 Hz), 6.86-6.88(d, 2H, J = 8.4 Hz), 7.26(d, 2H, J = 8.8 Hz), 7.58(d,
1
3
1
H, J = 16.0 Hz), 7.62(m, 1H), 7.77(s, 1H). C NMR (100 MHz, CDCl )
3
1
7

ACCEPTED MANUSCRIPT
δ 43.78, 51.85, 55.31, 111.43, 114.52, 114.52, 118.08, 118.16, 124.10,
1
1
5
3
26.10, 128.95, 128.95, 130.49, 137.93, 142.43, 151.56, 158.11, 159.60,
66.92, 182.77. ESI-MS: 352.1 [M+H]+, 374.1 [M+Na]+.
.1.1.10. (E)-methyl
-(1-(4-chlorobenzyl)-2,3-dioxoindolin-5-yl)acrylate (2i)
1
Yield 45%, purity 97%; H NMR (400 MHz, CDCl ) δ 3.80(s, 3H),
3
4
7
7
.92(s, 2H), 6.34-6.38(d, 1H, J = 16.0 Hz), 6.78-6.80(d, 1H, J = 8.0 Hz),
.27(d, 2H, J = 8.0 Hz), 7.33(d, 2H, J = 8.0 Hz), 7.59(d, 1H, J = 16.0 Hz),
1
3
.63(m, 1H), 7.79(s, 1H). C NMR (100 MHz, CDCl ) δ 43.64, 51.89,
3
111.22, 118.10, 118.41, 124.27, 128.84, 128.84, 129.39, 129.39, 130.79,
1
3
5
3
32.67, 134.37, 138.00, 142.27, 151.13, 158.09, 166.87, 182.37. ESI-MS:
56.1 [M+H]+, 378.1 [M+Na]+.
.1.1.11. (E)-methyl
-(1-(3,4-dichlorobenzyl)-2,3-dioxoindolin-5-yl)acrylate (2j)
1
Yield 39%, purity 95%; H NMR (400 MHz, CDCl ) δ 3.80(s, 3H),
3
4
7
.90(s, 2H), 6.35-6.39(d, 1H, J = 16.0 Hz), 6.78-6.80(d, 1H, J = 8.0 Hz),
.16-7.19(m, 1H), 7.43-7.45(d, 2H, J = 8.0 Hz), 7.58-7.62(d, 1H, J = 16.0
13
Hz), 7.65-7.67(m, 1H), 7.81(s, 1H). C NMR (100 MHz, CDCl ) δ 43.19,
3
5
1
1.90, 111.07, 118.12, 118.55, 124.38, 126.74, 129.38, 130.98, 131.19,
32.75, 133.42, 134.44, 138.09, 142.19, 150.82, 158.06, 166.83, 182.11.
ESI-MS: 390.1 [M+H]+, 412.1 [M+Na]+.
1
8

ACCEPTED MANUSCRIPT
5
3
.1.1.12. (E)-methyl
-(1-(4-bromobenzyl)-2,3-dioxoindolin-5-yl)acrylate (2h)
1
Yield 40%, purity 98%; H NMR (400 MHz, CDCl ) δ 3.80(s, 3H),
3
4
7
=
.90(s, 2H), 6.34-6.38(d, 1H, J = 16.0 Hz), 6.78-6.80(d, 1H, J = 8.0 Hz),
.20-7.22(d, 2H, J = 7.2 Hz), 7.48-7.50(d, 2H, J = 8.0 Hz), 7.59(d, 1H, J
13
16.0 Hz), 7.63(m, 1H), 7.79 (s, 1H). C NMR (100 MHz, CDCl ) δ
3
4
1
1
5
3
3.69, 51.88, 111.22, 118.10, 118.43, 122.42, 124.26, 129.14, 129.14,
30.80, 132.35, 132.35, 133.20, 138.00, 142.26, 151.11, 158.09, 166.85,
82.34. ESI-MS: 400.1 [M+H]+.
.1.1.13. (E)-methyl
-(1-(3-bromobenzyl)-2,3-dioxoindolin-5-yl)acrylate (2l)
1
Yield 38%, purity 99%; H NMR (400 MHz, CDCl ) δ 3.80(s, 3H),
3
4
7
1
1
1
.91(s, 2H), 6.35-6.39(m, 1H, J = 16.0 Hz), 6.78-6.80(d, 1H, J = 8.4 Hz),
.26(m, 2H), 7.46(m, 2H), 7.60(d, 1H, J = 16.0 Hz), 7.65(m, 1H), 7.81(s,
1
3
H). C NMR (100 MHz, CDCl ) δ 43.65, 51.89, 111.20, 118.11, 118.44,
3
23.25, 124.32, 126.00, 130.42, 130.75, 130.85, 131.64, 136.46, 138.08,
42.28, 151.07, 158.09, 166.87, 182.29. ESI-MS: 400.1 [M+H]+, 422.1
[M+Na]+.
5
3
.1.1.14. (E)-methyl
-(1-(2-bromobenzyl)-2,3-dioxoindolin-5-yl)acrylate (2m)
1
Yield 36%, purity 99%; H NMR (400 MHz, CDCl ) δ 3.80(s, 3H),
3
5
.07(s, 2H), 6.35-6.39(d, 1H, J = 16.0 Hz), 6.78-6.80(d, 1H, J = 8.0 Hz),
1
9

ACCEPTED MANUSCRIPT
7
2
.17-7.21(m, 2H), 7.25-7.29(m, 1H), 7.60(d, 1H, J = 16.0 Hz), 7.64(m,
13
H), 7.81(s, 1H). C NMR (100 MHz, CDCl ) δ 44.23, 51.88, 111.62,
3
1
18.10, 118.39, 122.88, 124.22, 128.12, 128.15, 129.80, 130.83, 132.98,
33.37, 138.17, 142.34, 151.23, 158.27, 166.88, 182.37. ESI-MS: 400.1
M+H]+.
1
[
5
3
.1.1.15. (E)-methyl
-(1-(naphthalen-2-ylmethyl)-2,3-dioxoindolin-5-yl)acrylate (2n)
1
Yield 44%, purity 97%; H NMR (400 MHz, CDCl ) δ 3.79(s, 3H),
3
5
7
.12(s, 2H), 6.31-6.35(d, 1H, J = 16.0 Hz), 6.84-6.86(d, 1H, J = 8.4 Hz),
.40-7.42(m, 1H), 7.48-7.51(m, 2H), 7.54-7.58(m, 2H), 7.79-7.85(m, 5H).
1
3
C NMR (100 MHz, CDCl ) δ 44.52, 51.86, 111.51, 118.10, 118.23,
3
1
1
24.17, 124.90, 126.51, 126.52, 126.74, 127.75, 127.82, 129.29, 130.61,
31.53, 133.05, 133.26, 138.01, 142.38, 151.49, 158.24, 166.91, 182.65.
ESI-MS: 372.2 [M+H]+, 394.2 [M+Na]+.
5
3
.1.1.16. (E)-methyl
-(2,3-dioxo-1-(2-phenoxyethyl)indolin-5-yl)acrylate (2o)
1
Yield 45%, purity 100%; H NMR (400 MHz, CDCl ) δ 3.81(s, 3H),
3
4
6
7
.14-4.17(t, 2H), 4.25-4.28(t, 2H), 6.37-6.41(d, 1H, J = 16.0 Hz),
.80-6.82(d, 2H, J = 8.0 Hz), 6.94-6.97(t, 1H), 7.21-7.27(m, 3H),
1
3
.61-7.65(d, 1H, J = 16.0 Hz), 7.74-7.76(t, 2H). C NMR (100 MHz,
CDCl ) δ 40.46, 51.87, 65.70, 111.78, 114.30, 114.30, 117.92, 118.14,
3
2
0

ACCEPTED MANUSCRIPT
21.57, 124.03, 129.63, 129.63, 130.46, 137.89, 142.48, 152.34, 157.87,
58.36, 166.96, 182.53. ESI-MS: 352.2 [M+H]+, 374.2 [M+Na]+.
.1.1.17. (E)-methyl
1
1
5
3
-(1-(3,5-dimethylbenzyl)-2,3-dioxoindolin-5-yl)acrylate (2p)
1
Yield 42%, purity 97%; H NMR (400 MHz, CDCl ) δ 2.28(s, 6H),
3
3
.80(s, 3H), 4.86(s, 2H), 6.33-6.37(d, 1H, J = 16.0 Hz), 6.82-6.84(d, 1H,
J = 8.4 Hz), 6.92-6.94(m, 3H), 7.59(d, 1H, J = 16.0 Hz), 7.62(m, 1H),
1
3
7
.78(s, 1H). C NMR (100 MHz, CDCl ) δ 21.26, 21.26, 44.24, 51.86,
3
111.50, 118.06, 118.12, 124.10, 125.19, 125.19, 125.19, 130.00, 130.48,
1
3
5
34.02, 138.03, 138.84, 142.48, 151.68, 158.17, 166.94, 182.80. ESI-MS:
50.2 [M+H]+, 372.2 [M+Na]+.
.1.2. Synthesis of compounds 2q-2s and 3a-3o.
To a microwave reactor vial (5 ml) which contained the solution of 4 or 5
or 6 or 7-bromoindoline-2,3-dione (1.0 g, 4.4 mmol) in DMF (5 ml) were
added PdCl (PPh ) (155 mg, 0.22 mmol), CH COOK (0.6 g, 6.2 mmol)
2
3 2
3
and ethenyl (5.3 mmol) under the atmosphere of argon. The microwave
reactor vial was caped and placed into the microwave cavity. The reaction
mixture was irradiated at high level for 30 min at 145 ℃. The reaction
mixture was cooled to room temperature, pourd onto 100 ml ice-water
and then extracted with dichloromethane (3×100 ml). The combined
organic layers were washed with water (2×100 ml), brine (100 ml) and
dried over anhydrous magnesium sulfate. The solvent was removed under
2
1

ACCEPTED MANUSCRIPT
vacuum to afford the crude which was purified by flash column
chromatography (silica gel, petroleum ether/ethyl acetate 3:1) to yield the
desired (2-Carboxyethenyl) isatin derivatives.
To a flask (25 ml) which contained the solution of (2-Carboxyethenyl)
isatin derivatives (0.86 mmol) in dry N, N-dimethylformamide (2 ml).
The reaction temperature was maintained at 0 ℃ followed by addition
K CO (2.6 mmol). Stirring 5 min, was added alkyl or benzyl (1.0 mmol)
2
3
and the mixture allowed warm to room temperature. The reaction mixture
was stirred at room temperature for 4 h. The orange solution was poured
water (25 ml) and extracted with dichloromethane (3 × 50 ml). The
combined extracts were dried over anhydrous magnesium sulfate, filtered
and evaporated. Chromatography (petroleum ether/ethyl acetate 8:1)
afforded the title compounds 2q-2s and 3a-3o.
5
3
.1.2. 1. (E)-methyl
-(1-(4-methoxybenzyl)-2,3-dioxoindolin-4-yl)acrylate (2q)
1
Yield 33%, purity 98%; H NMR (400 MHz, CDCl ) δ 3.78(s, 3H),
3
3
.84(s, 3H), 4.87(s, 2H), 6.61-6.65(d, 1H, J = 16.0 Hz), 6.79-6.81(d, 1H,
J = 7.6 Hz), 6.85-6.88(m, 2H), 7.25-7.30(m, 3H), 7.43-7.47(t, 1H),
1
3
8
5
1
1
.39-8.43(d, 1H, J = 16.0 Hz). C NMR (100 MHz, CDCl ) δ 43.62,
3
2.07, 55.30, 111.68, 114.46, 114.46, 114.83, 121.12, 124.04, 126.27,
28.89, 128.89, 135.11, 137.68, 137.73, 151.07, 157.52, 159.51, 166.41,
83.02. ESI-MS: 352.2 [M+H]+, 374.1 [M+Na]+.
2
2

ACCEPTED MANUSCRIPT
5
3
.1.2. 2. (E)-methyl
-(1-(4-methoxybenzyl)-2,3-dioxoindolin-6-yl)acrylate (2r)
1
Yield 36%, purity 98%; H NMR (400 MHz, CDCl ) δ 3.79(s, 3H),
3
3
7
.82(s, 3H), 4.89(s, 2H), 6.41-6.45(d, 1H, J = 16.0 Hz), 6.88-6.90(m, 3H),
.21-7.23(d, 1H, J = 7.6 Hz), 7.26-7.29(m, 2H), 7.54-7.58(d, 1H, J = 16.0
13
Hz), 7.60-7.62(d, 1H, J = 7.6 Hz). C NMR (100 MHz, CDCl ) δ 43.65,
3
5
1
1
5
3
2.11, 55.31, 109.65, 114.57, 114.57, 118.53, 122.19, 123.61, 125.77,
26.13, 128.88, 128.88, 142.74, 143.85, 151.12, 158.22, 159.58, 166.41,
82.53. ESI-MS: 352.2 [M+H]+, 374.1 [M+Na]+.
.1.2. 3. (E)-methyl
-(1-(4-methoxybenzyl)-2,3-dioxoindolin-7-yl)acrylate (2s)
1
Yield 34%, purity 100%; H NMR (400 MHz, CDCl ) δ 3.78(s, 3H),
3
3
7
7
.80(s, 3H), 5.09(s, 2H), 6.16-6.20(d, 1H, J = 15.6 Hz), 6.87-6.89(m, 2H),
.12-7.15(t, 1H), 7.21-7.23(d, 2H, J = 8.8 Hz), 7.53-7.55(m, 1H),
1
3
.67-7.69(m, 1H), 7.82-7.86(d, 1H, J = 15.6 Hz). C NMR (100 MHz,
CDCl ) δ 45.66, 51.89, 55.29, 114.44, 114.44, 119.18, 120.94, 122.72,
3
1
1
5
3
24.30, 126.53, 126.63, 127.77, 127.77, 137.86, 139.65, 148.18, 159.25,
59.44, 166.15, 182.79. ESI-MS: 374.2 [M+Na]+.
.1.2. 4. (E)-ethyl
-(1-(4-methoxybenzyl)-2,3-dioxoindolin-5-yl)acrylate (3a)
1
Yield 41%, purity 100%; H NMR (400 MHz, CDCl ) δ 1.31-1.34(t, 3H),
3
3
.78(s, 3H), 4.23-4.28(m, 2H), 4.88(s, 2H), 6.32-6.36(d, 1H, J = 16.0 Hz),
2
3

ACCEPTED MANUSCRIPT
6
1
.82-6.84(d, 1H, J = 8.4 Hz), 6.86-6.89(m, 2H), 7.25-7.27(m, 2H), 7.57(d,
1
3
H, J = 16.0 Hz), 7.61(m, 1H), 7.77(s, 1H). C NMR (100 MHz, CDCl )
3
δ 14.28, 43.78, 55.31, 60.74, 111.40, 114.51, 114.51, 118.08, 118.66,
1
1
5
24.11, 126.11, 128.95, 128.95, 130.60, 137.89, 142.14, 151.50, 158.13,
59.59, 166.51, 182.79. ESI-MS: 366.2 [M+H]+.
.1.2. 5. (E)-ethyl 3-(1-(4-bromobenzyl)-2,3-dioxoindolin-5-yl)acrylate
(3b)
1
Yield 32%, purity 98%; H NMR (400 MHz, CDCl ) δ 1.31-1.34(t, 3H),
3
4
1
.23-4.28(m, 2H), 4.90(s, 2H), 6.33-6.37(d, 1H, J = 16.0 Hz), 6.77-6.79(d,
H, J = 8.4 Hz), 7.20-7.22(d, 2H, J = 8.4 Hz), 7.48-7.50(d, 2H, J = 8.4
1
3
Hz), 7.55-7.59(d, 1H, J = 16.0 Hz), 7.62-7.64(m, 1H), 7.79(s, 1H). C
NMR (100 MHz, CDCl ) δ 14.27, 43.68, 60.77, 111.22, 118.08, 118.88,
3
1
1
5
3
22.39, 124.25, 129.15, 129.15, 130.88, 132.32, 132.32, 133.23, 137.98,
42.00, 151.05, 158.12, 166.46, 182.37. ESI-MS: 414.1 [M+H]+.
.1.2. 6. (E)-butyl
-(1-(4-methoxybenzyl)-2,3-dioxoindolin-5-yl)acrylate (3c)
1
Yield 44%, purity 96%; H NMR (400 MHz, CDCl ) δ 0.94-0.97(t, 3H),
3
1
2
6
1
4
.40-1.45(m, 2H), 1.64-1.69(m, 2H), 3.78(s, 3H), 4.18-4.21(t, 2H), 4.88(s,
H), 6.33-6.37(d, 1H, J = 16.0 Hz), 6.82-6.84(d, 1H, J = 8.4 Hz),
.86-6.89(m, 2H), 7.25-7.27(m, 2H), 7.56(d, 1H, J = 16.0 Hz), 7.62(m,
1
3
H), 7.77 (s, 1H). C NMR (100 MHz, CDCl ) δ 13.72, 19.17, 30.71,
3
3.74, 55.29, 64.63, 111.42, 114.49, 114.49, 118.06, 118.64, 124.08,
2
4

ACCEPTED MANUSCRIPT
26.14, 128.96, 128.96, 130.57, 137.91, 142.11, 151.49, 158.13, 159.57,
66.58, 182.80. ESI-MS: 394.2 [M+H]+.
1
1
5
3
.1.2. 7. (E)-butyl
-(1-(4-bromobenzyl)-2,3-dioxoindolin-5-yl)acrylate (3d)
1
Yield 33%, purity 98%; H NMR (400 MHz, CDCl ) δ 0.94-0.97(t, 3H),
3
1
6
2
.39-1.45(m, 2H), 1.64-1.70(m, 2H), 4.18-4.22(t, 2H), 4.90(s, 2H),
.34-6.38(d, 1H, J = 16.0 Hz), 6.76-6.78(d, 1H, J = 8.0 Hz), 7.20-7.22(d,
H, J = 8.4 Hz), 7.48-7.50(d, 2H, J = 8.4 Hz), 7.55-7.59(d, 1H, J = 16.0
13
Hz), 7.62-7.64(m, 1H), 7.80(s, 1H). C NMR (100 MHz, CDCl ) δ 13.73,
3
1
1
1
5
3
9.17, 30.71, 43.68, 64.68, 111.22, 118.08, 118.91, 122.39, 124.24,
29.15, 129.15, 130.89, 132.32, 132.32, 133.24, 137.98, 141.95, 151.04,
58.11, 166.52, 182.37. ESI-MS: 442.1 [M+H]+.
.1.2.8. (E)-tert-butyl
-(1-(4-methoxybenzyl)-2,3-dioxoindolin-5-yl)acrylate (3e)
1
Yield 41%, purity 97%; H NMR (400 MHz, CDCl ) δ 1.52(s, 9H),
3
3
.78(s, 3H), 4.88(s, 2H), 6.26-6.30(d, 1H, J = 16.0 Hz), 6.81-6.83(d, 1H,
J = 8.4 Hz), 6.86-6.88(m, 2H), 7.25-7.27(m, 2H), 7.45-7.49(d, 1H, J =
1
3
1
2
1
1
6.0 Hz), 7.58-7.60(m, 1H), 7.75 (s, 1H). C NMR (100 MHz, CDCl ) δ
3
8.15, 28.15, 28.15, 43.74, 55.30, 80.89, 111.36, 114.49, 114.49, 118.06,
20.59, 124.00, 126.18, 128.96, 128.96, 130.81, 137.78, 141.09, 151.32,
58.15, 159.57, 165.78, 182.85. ESI-MS: 394.3 [M+H]+, 416.2
[M+Na]+.
2
5

ACCEPTED MANUSCRIPT
.1.2. 9. (E)-tert-butyl
5
3
-(1-(4-bromobenzyl)-2,3-dioxoindolin-5-yl)acrylate (3f)
1
Yield 34%, purity 98%; H NMR (400 MHz, CDCl ) δ 1.52(s, 9H),
3
4
7
7
2
1
1
5
3
.89(s, 2H), 6.27-6.31(d, 1H, J = 16.0 Hz), 6.75-6.77(d, 1H, J = 8.4 Hz),
.20-7.22(d, 2H, J = 8.4 Hz), 7.47(d, 1H, J = 16.0 Hz), 7.49(m, 2H),
1
3
.59-7.62(m, 1H), 7.78(s, 1H). C NMR (100 MHz, CDCl ) δ 28.15,
3
8.15, 28.15, 43.68, 80.98, 111.13, 118.08, 120.88, 122.41, 124.18,
29.15, 129.15, 131.14, 132.34, 132.34, 133.25, 137.84, 140.92, 150.86,
58.13, 165.73, 182.41. ESI-MS: 442.1 [M+H]+.
.1.2. 10. (E)-2-hydroxyethyl
-(1-(4-methoxybenzyl)-2,3-dioxoindolin-5-yl)acrylate (3g)
1
Yield 43%, purity 99%; H NMR (400 MHz, CDCl ) δ 3.78(s, 3H),
3
3
1
1
4
1
1
5
3
.89-3.91(m, 2H), 4.34-4.36(m, 2H), 4.89(s, 2H), 6.37-6.41(d, 1H, J =
6.0 Hz), 6.84(d, 1H, J = 8.0 Hz), 6.88(m, 2H), 7.27 (m, 2H), 7.61(m,
1
3
H), 7.62(d, 1H, J = 16.0 Hz) 7.78(s, 1H). C NMR (100 MHz, CDCl ) δ
3
3.79, 55.32, 58.47, 66.39, 111.46, 114.52, 114.52, 117.90, 118.10,
24.17, 126.07, 128.96, 128.96, 130.36, 138.01, 143.01, 151.67, 158.11,
59.60, 166.77, 182.73. ESI-MS: 382.2 [M+H]+, 404.2 [M+Na]+.
.1.2. 11. (E)-2-hydroxyethyl
-(1-(4-bromobenzyl)-2,3-dioxoindolin-5-yl)acrylate (3h)
1
Yield 27%, purity 97%; H NMR (400 MHz, DMSO-d6) δ 3.61-3.62(t,
2
H), 4.13-4.15(t, 2H), 4.91(s, 2H), 6.63-6.67(d, 1H, J = 16.0 Hz),
2
6

ACCEPTED MANUSCRIPT
6
2
1
.98-7.00(d, 1H, J = 8.4 Hz), 7.40-7.43(d, 2H, J = 8.4 Hz), 7.52-7.55(d,
H, J = 8.4 Hz), 7.64-7.68(d, 1H, J = 16.0 Hz), 7.93-7.96 (m, 1H) 7.98(s,
1
3
H). C NMR (100 MHz, CDCl ) δ 42.92, 59.50, 66.34, 111.76, 117.97,
3
118.91, 121.21, 124.52, 129.86, 130.01, 130.12, 131.97, 132.06, 135.26,
1
4
5
38.13, 143.59, 151.58, 159.07, 166.79, 182.83. ESI-MS: 430.1 [M+H]+,
52.1 [M+Na]+.
.1.2. 12. (E)-3-(1-(4-methoxybenzyl)-2,3-dioxoindolin-5-yl)acrylic
acid (3i)
Yield 45%, purity 100%; H NMR (400 MHz, DMSO-d6) δ 3.72(s, 3H),
1
4
7
1
.85(s, 2H), 6.50-6.54(d, 1H, J = 16.0 Hz), 6.88-6.90(d, 2H, J = 8.8 Hz),
.01-7.03(d, 1H, J = 8.8 Hz), 7.36-7.38(d, 2H, J = 8.8 Hz), 7.55-7.59(d,
13
H, J = 16.0 Hz), 7.90-7.92(m, 2H), 12.35(s, 1H). C NMR (100 MHz,
DMSO-d6) δ 42.39, 54.98, 111.31, 113.93, 113.93, 118.19, 118.49,
1
1
5
23.81, 127.06, 128.80, 128.80, 129.54, 137.40, 142.46, 151.10, 158.37,
58.63, 167.43, 182.56. ESI-MS: 338.2 [M+H]+.
.1.2. 13.
(E)-3-(1-(4-methoxybenzyl)-2,3-dioxoindolin-5-yl)acrylamide (3j)
1
Yield 44%, purity 100%; H NMR (400 MHz, DMSO-d6) δ 3.72(s, 3H),
4
7
7
.85(s, 2H), 6.55-6.59(d, 1H, J = 16.0 Hz), 6.88-6.90(d, 2H, J = 8.8 Hz),
.01-7.03(d, 1H, J = 8.4 Hz), 7.08(s, 1H), 7.36(m, 2H), 7.41(s, 2H),
1
3
.76(d, 1H, J = 16.0 Hz), 7.77(m, 1H). C NMR (100 MHz, DMSO-d6)
δ 42.99, 55.55, 111.95, 114.51, 114.51, 118.77, 122.29, 123.15, 127.65,
2
7

ACCEPTED MANUSCRIPT
1
1
5
29.36, 129.36, 130.77, 137.42, 138.19, 151.08, 158.92, 159.21, 166.98,
83.27. ESI-MS: 337.2 [M+H]+, 359.1 [M+Na]+.
.1.2. 14. (E)-3-(1-(4-bromobenzyl)-2,3-dioxoindolin-5-yl)acrylamide
(3k)
1
Yield 30%, purity 97%; H NMR (400 MHz, DMSO-d6) δ 4.90(s, 2H),
6
7
7
.55-6.59(d, 1H, J = 16.0 Hz), 6.97-7.00(d, 1H, J = 8.4 Hz), 7.08(s, 1H),
.39 (d, 1H, J = 16.0 Hz), 7.41(m, 3H), 7.53-7.55(d, 2H, J = 8.4 Hz),
1
3
.75-7.81(m, 2H). C NMR (100 MHz, DMSO-d6) δ 42.91, 111.80,
18.92, 121.18, 122.33, 123.19, 130.14, 130.14, 130.86, 131.95, 131.95,
35.37, 137.35, 138.18, 150.84, 159.06, 166.98, 183.01. ESI-MS: 385.0
M+H]+, 407.0 [M+Na]+.
.1.2.15.
E)-1-(4-methoxybenzyl)-5-(3-morpholino-3-oxoprop-1-en-1-yl)indoli
ne-2,3-dione(3l)
Yield 39%, purity 99%; H NMR (400 MHz, CDCl ) δ 3.69-3.71(m, 8H),
1
1
[
5
(
1
3
3
.78(s, 3H), 4.88(s, 2H), 6.76-6.80(d, 1H, J = 15.2 Hz), 6.81-6.83(d, 1H,
J = 8.0 Hz), 6.86-6.88(d, 2H, J = 8.8 Hz), 7.26(d, 2H, J = 8 .4Hz), 7.57(m,
1
3
1
H), 7.60(d, 1H, J = 15.2 Hz), 7.82(s, 1H). C NMR (100 MHz, CDCl )
3
δ 42.41, 43.79, 46.20, 55.31, 58.46, 66.83, 111.36, 114.50, 114.50, 116.93,
18.08, 123.14, 126.13, 128.98, 128.98, 131.30, 138.49, 140.94, 151.08,
58.16, 159.59, 165.43, 183.08. ESI-MS: 407.2 [M+H]+.
1
1
2
8