Synthesis and evaluation of 3-ylideneoxindole acetamides as potent anticancer agents

Article abstract of DOI:10.1016/j.ejmech.2015.04.062

Indirubin, an active component in the traditional Chinese medicine formula Danggui Longhui Wan, shows promising anticancer effects. Meisoindigo is an analog derived from indirubin, which is less toxic and appears to be even more potent against cancer. In considering meisoindigo as a structural template for the development of new drugs, we designed and synthesized a series of 3-ylideneoxindole acetamides as novel anticancer agents. The acetamides were then evaluated for in vitro and in vivo anticancer activities. The 3-ylideneoxindole acetamides were found to have better anticancer activity than was indirubin-3'-oxime in several cancer cell lines and also displayed a spectrum of activity similar to that of the drug candidate roscovitine, a CDK inhibitor. Among the 3-ylideneoxindole acetamides, compound 10 showed particularly good efficacy. Cell cycle analysis further revealed that compound 10 arrested cells in the G1 phase and caused an increase in the sub-G1 population, indicating that the apoptosis pathway had been induced. In addition, exposure of cells to compound 10 led to the upregulation of the cell-cycle regulator cyclin D1, which was sustained at a high level. In contrast, the same compound induced a short-term elevation in the level of cyclin E, which was followed by a rapid decrease and the attenuation of Rb phosphorylation. Furthermore, a docking model suggests that compound 10 binds to the active site of CDK4. In testing the therapeutic potency of compound 10 on CT26-xenografted BALB/c mice, a significant reduction in tumor size comparable to that of cisplatin was found when administrated via the i.p. route. The mice presented no loss of body weight, indicating that this compound possesses low toxicity. In the future, we are planning in vivo investigations of these new active anticancer agents to better elucidate active mechanisms at the cellular level and thus benefit the development of anticancer therapies.

Full text of DOI:10.1016/j.ejmech.2015.04.062

Accepted Manuscript
Synthesis and evaluation of 3-ylideneoxindole acetamides as potent anticancer
agents
Chun-Tang Chiou, Wei-Chun Lee, Jiahn-Haur Liao, Jing-Jy Cheng, Lie-Chwen Lin,
Chih-Yu Chen, Jen-Shin Song, Ming-Hsien Wu, Kak-Shan Shia, Wen-Tai Li
PII:
S0223-5234(15)30025-8
10.1016/j.ejmech.2015.04.062
EJMECH 7877
DOI:
Reference:
To appear in: European Journal of Medicinal Chemistry
Received Date: 9 March 2015
Revised Date: 28 April 2015
Accepted Date: 29 April 2015
Please cite this article as: C.-T. Chiou, W.-C. Lee, J.-H. Liao, J.-J. Cheng, L.-C. Lin, C.-Y. Chen, J.-
S. Song, M.-H. Wu, K.-S. Shia, W.-T. Li, Synthesis and evaluation of 3-ylideneoxindole acetamides
as potent anticancer agents, European Journal of Medicinal Chemistry (2015), doi: 10.1016/
j.ejmech.2015.04.062.
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to
our customers we are providing this early version of the manuscript. The manuscript will undergo
copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please
note that during the production process errors may be discovered which could affect the content, and all
legal disclaimers that apply to the journal pertain.

ACCEPTED MANUSCRIPT
IC50 (µM)
KB
Compd
MCF-7
2.3±1.8
Hep3B
2.7±1.6
SF-268
3.6±3.5
MKN-45
3.6±0.3
1
0
2.2±1.9

ACCEPTED MANUSCRIPT
1
2
3
Synthesis and Evaluation of 3-Ylideneoxindole Acetamides as Potent
Anticancer Agents
a
a
b
a
a
4
5
6
7
8
9
0
1
2
Chun-Tang Chiou , Wei-Chun Lee , Jiahn-Haur Liao , Jing-JyCheng , Lie-Chwen Lin , Chih-Yu
a
c
c
c
a
Chen , Jen-Shin Song , Ming-Hsien Wu , Kak-Shan Shia , and Wen-Tai Li *
a
National Research Institute of Chinese Medicine, Ministry of Health and Welfare
Taipei 11221, Taiwan
b
Institute of Biological Chemistry, Academia Sinica
1
1
1
Taipei 11529, Taiwan
c
Institute of Biotechnology and Pharmaceutical Research, National Health Research Institutes
Miaoli 35053, Taiwan
13
14
15
16
17
18
19
20
21
*
Correspondence to:
Wen-Tai Li, Ph.D.
National Research Institute of Chinese Medicine, Ministry of Health and Welfare
155-1, Sec. 2, Linong St., Beitou Distrct,
Taipei 11221, Taiwan, ROC
Tel: +886 2 2820 1999 ext. 8261;
Fax: +886 2 2825 0743;
E-mail: wtli@nricm.edu.tw
1

ACCEPTED MANUSCRIPT
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
Abstract
Indirubin, an active component in the traditional Chinese medicine formula Danggui Longhui
Wan, shows promising anticancer effects. Meisoindigo is an analogue derived from indirubin,
which is less toxic and appears to be even more potent against cancer. In considering meisoindigo
as a structural template for the development of new drugs, we designed and synthesized a series of
3-ylideneoxindole acetamides as novel anticancer agents. The acetamides were then evaluated for
in vitro and in vivo anticancer activities. The 3-ylideneoxindole acetamides were found to have
better anticancer activity than was indirubin-3'-oxime in several cancer cell lines and also
displayed a spectrum of activity similar to that of the drug candidate roscovitine, a CDK inhibitor.
Among the 3-ylideneoxindole acetamides, compound 10 showed particularly good efficacy. Cell
cycle analysis further revealed that compound 10 arrested cells in the G1 phase and caused an
increase in the sub-G1 population, indicating that the apoptosis pathway had been induced. In
addition, exposure of cells to compound 10 led to the upregulation of the cell-cycle regulator
cyclin D1, which was sustained at a high level. In contrast, the same compound induced a
short-term elevation in the level of cyclin E, which was followed by a rapid decrease and the
attenuation of Rb phosphorylation. Furthermore, a docking model suggests that compound 10
binds to the active site of CDK4. In testing the therapeutic potency of compound 10 on
CT26-xenografted BALB/c mice, a significant reduction in tumor size comparable to that of
cisplatin was found when administrated via the i.p. route. The mice presented no loss of body
weight, indicating that this compound possesses low toxicity. In the future, we are planning in
vivo investigations of these new active anticancer agents to better elucidate active mechanisms at
the cellular level and thus benefit the development of anticancer therapies.
2

ACCEPTED MANUSCRIPT
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
Keywords: Danggui Longhui Wan; Meisoindigo; 3-ylideneoxindole acetamide; anticancer activity.
3

ACCEPTED MANUSCRIPT
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
1. Introduction
Herbal plants used in traditional Chinese medicine provide valuable sources of compounds
for the development of new drugs. Compounds used in the prevention and treatment of cancer
have attracted particular attention. Several Chinese herbs, such as Camptotheca acuminate,
Schisandra chinensis, Panax ginseng, and Ziziphus jujube have demonstrated the ability to induce
cell cycle arrest and play an important role in cancer prevention [1-4]. The traditional Chinese
herbal formula Danggui Longhui Wan is a combination of 11 herbal medicines used to treat
chronic myelocytic leukaemia (CML). The Institute of Hematology at the Chinese Academy of
Medical Sciences has attributed the antileukemic activities of Danggui Longhui Wan to the
presence of indirubin, which has been shown to inhibit cyclin-dependent kinase (CDK) activity,
resulting in cell cycle arrest [5-6]. Furthermore, long-term animal studies have shown that
indirubin does not possess the toxicity of conventional anticancer drugs and does not affect bone
marrow or the production of hematopoietic stem cells [7]. Several analogues have been developed
from indirubin or identified in Qing Dai. These analogues include indirubin-3’-monoxime,
TM
5-halogen indirubin, methylisoindigo, and Natura , all of which exhibit potent antitumor
activities in animal models [8-10].
Cyclin-dependent kinases (CDKs) play crucial roles in cell growth by initiating and
regulating the cycle of cell-division. Cancer cell growth is highly associated with CDKs and their
regulators; therefore inhibitors of CDKs may serve as anti-cancer agents. Previous results have
suggested that cell differentiation may be a potential target for novel anticancer agents due to the
participation of CDK2 and CDK4/6 [11-12]. In addition, the 3-alkylidene oxindole molecule is a
key substructure in many protein kinase inhibitors, such as BIBF1120, GW8510, and the Aurora B
inhibitor, hesperidin [13-15].
4

ACCEPTED MANUSCRIPT
91
92
93
94
95
96
97
98
99
While searching for anticancer lead compounds from plants used in traditional Chinese
medicine and looking for novel structures worth to develop. Compounds such as indirubin and
meisoindigo are worthy of further investigation, due to the relatively potent anticancer activities
and low toxicity. In this study, we focused on meisoindigo, which has higher anticancer activity
and fewer side effects than indirubin, as a template for new anticancer agents.
The 3-alkylidene oxindole molecule is referred to as a ‘‘privileged structure’’ due to its
capacity to bind CDKs with high affinity. Meisoindigo contains two oxindole rings, and in this
study, we investigated the effect that opening the second ring has on anticancer activity. The
ring-opening structure is usually more flexible than the ring system, which makes it easy to
100 incorporate a variety of functional groups in order to increase bioactivity or solubility. Herein, we
101 describe the synthesis and biological evaluation of 3-ylideneoxindole acetamides, novel anticancer
102 agents designed through the modification of meisoindigo (Chart 1).
103
104
105 Chart 1. Meisoindigo and the template used for structural modification
1
1
1
06
07 2. Results and Discussion
08 2.1. Chemistry
5

ACCEPTED MANUSCRIPT
1
1
1
1
1
1
1
1
1
1
1
1
09
The general method for the synthesis of 3-ylideneoxindole acetamides is shown in Scheme 1.
10 Commercially available isatins were used as starting materials, and the other substituted isatins
11 were synthesized according to Sandmeyer’s method. The synthesis of isatin 1 began by treating
12 para-substituted anilines with chloral hydrate and hydroxylamine hydrochloride under acidic
13 conditions [16]. A substitution reaction with isatin 1 and aryl halides produced good yields of
14 N-substituted isatin 2. Coupling reaction of diethylphosphonoacetic acid and amines was then
15 performed to synthesize diethylphosphonoacetic amide 3. Nucleophilic addition of isatin 2 at the
16 3-position of diethylphosphonoacetic amide
3
followed by elimination (i.e. the
17 Horner-Wadsworth-Emmons reaction) was proposed to generate satisfactory yields of the desired
18 3-ylideneoxindole acetamide (4-39). It is worth noting that most 3-ylideneoxindole acetamides
19 obtained using this method was (E)-configuration and the (E)-configuration was identified by
20 NOE experiments.
O
O
R1
R1
R1
chloral hydrate, Na SO
Aryl halide
NaH, DMF
2
4
O
O
N
.
N
NH OH HCl, HCl, H O
NH2
2
2
H
R2
1
2
O
O
R'
R'
O
O
P
EDCI, DMAP
DIPEA, DMF
P
+
H N
2
HO
O
O
N
O
O
H
O
3
R'
HN
R1
O
O
R'
K CO
2 3
O
O
+
P
O
R1
N
THF, H O
2
N
H
O
O
R2
N
2
3
R2
4-39
121
6

ACCEPTED MANUSCRIPT
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
22 Scheme 1. Synthesis of 3-ylideneoxindole acetamides
23
24 2.2. In vitro anticancer activity against human cancer cell lines
25
All of the synthesized 3-ylideneoxindole acetamides were initially evaluated for anticancer
26 activity using human NCI-H460 lung cancer cells. We studied structure-activity relationships
27 (SARs) for compounds containing various substitutions in the 3-ylideneoxindole acetamide
1
28 scaffold (Table 1). For this, 3-ylideneoxindole acetamides with a hydrogen group at R and
2
3
2
29 different substituted groups at R and R were first synthesized; we then assessed the effects of R
3
30 and R positions on the anticancer activities of 3-ylideneoxindole acetamides. With the same
2
31 substitution of the 4-chlorophenyl group at the R position, compounds 4 and 5 (with p-pyridinyl
3
32 and 6-quinolinyl groups at R , respectively) presented nearly equal potency against NCI-H460
33 cells. A comparison of compounds 4, 5, 7, and 8, suggested that the 4-chlorophenyl and
2
34 3-methylisoxazolyl groups at the R position have similar potency against NCI-H460 cancer cells.
2
35 Thus, we fixed the 3-methylisoxazolyl group at the R position and compared the anticancer
3
36 activities of different R amino groups. The electron donating substituents, such as methoxy
3
37 (compound 6) and hydroxyl (compound 9) groups, on para-phenyl in the R position presented
38 potent anticancer activity against NCI-H460 cells. Compounds 6, 7, and 8, which respectively
3
39 contain a para-methoxyphenyl, para-pyridinyl, or 6-quinolinyl group in the R position, were
40 found to have similar anticancer activities. However, a decrease in anticancer activity was
3
41 observed when iodo was substituted in the 2- or 3-position of the phenyl group at R . Compound
3
42 14, with an electron withdrawing nitro group on para-phenyl in the R position, did not present
43 anticancer activity, even at a concentration of 100 µM. Compound 13, with a phenyl group in the
7

ACCEPTED MANUSCRIPT
3
3
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
44 R position, presented poor anticancer activity. Conversely, the heteroaryl groups in the R position,
45 such as p-pyridinyl (compound 7), 6-quinolinyl (compound 8), and 3-methylisoxazolyl
46 (compound 29), showed good anticancer activities, ranked by potency as follows: 6-quinolinyl >
3
47 p-pyridinyl > 3-methylisoxazolyl. As 6-quinolinyl in the R position of 3-ylideneoxindole
48 acetamide presented the most potent anticancer activity, we continued our investigation by
1
2
49 focusing on various substitutions in the R and R positions while maintaining 6-quinolinyl in the
3
2
3
50 R position. When R was an alkyl or hydrogen and R was a 6-quinolinyl group, anticancer
51 activity against NCI-H460 was poor. Compounds 15, 17, 18, and 22, with a substituent of
2
3
52 para-phenyl in the R position and a 6-quinolinyl group in the R position, demonstrated potent
53 activities against NCI-H460 cancer cells with an IC of approximately 1 µM. Compounds 23 and
5
0
2
54 5, which possess a substituent of meta-phenyl in the R position comparable to the position of
55 para-phenyl on 3-ylideneoxindole acetamides, led to a slight decrease in anti-cancer activity.
2
56 Moreover, compound 28, with a 4-methoxyphenyl group in the R position, was found to be less
57 active than other compounds that possessed 4-halophenyl (5 and 18), 4-nitrophenyl (22), or
2
58 4-methyl benzoate (15) at R . This indicates that the electron deficiency associated with
2
59 para-phenyl at the R position is important to the cytotoxic activity of 3-ylideneoxindole
1
60 acetamides. Furthermore, compared with other substituents at R , the anticancer activity of
61 methoxy (compound 27) is superior to that of methyl (compound 24) and halo (compound 20).
62
Further evaluations were conducted on selected active 3-ylideneoxindole acetamides using
63 five cancer cell lines: human breast adenocarcinoma MCF-7, human hepatocellular Hep3B, human
64 epidermoid carcinoma KB, human central nervous system cancer SF-268 and human gastric
65 cancer MKN-45 (Table 2). Most of the 3-ylideneoxindole acetamides exhibited a wide spectrum
8

ACCEPTED MANUSCRIPT
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
66 of anticancer activities against human cancer cells and were found to be more active than
67 indirubin-3’-oxime and roscovitine.
68
69 2.3. Lipophilicity
70
The partition coefficient (ClogP) is used to estimate the lipophilicity and solubility of
71 chemical compounds, where a higher ClogP value indicates greater lipophilicity and poor aqueous
72 solubility. We calculated ClogP values for various 3-ylideneoxindole acetamides (Table 1) and
73 determined that the 3-methylisoxazolyl ring provides lower lipophilicity than does a phenyl ring
2
74 when located at the R position of 3-ylideneoxindole acetamides [17]. Conversely, 6-quinolinyl at
3
75 R showed higher ClogP values than did other phenyl substituents in the same position. ClogP
76 values for compound 10, indirubin-3’-oxime and roscovitine are predicted to be 3.29, 2.40 and
77 3.69, respectively. However, we did not identify a correlation between ClogP values and in vitro
78 anticancer activity.
79
80 2.4. 3-Ylideneoxindole acetamide arrested cell cycle progression in the early G1 phase
81
To assess the ability of 3-ylideneoxindole acetamide to induce cell cycle arrest and to
82 determine the point of arrest in cancer cells, we evaluated the antiproliferative mechanisms of
83 compound 10 on A549 lung cancer cells. Flow cytometry revealed that compound 10 arrested
84 cells in the G1 phase and induced an increase in the sub-G1 population via apoptosis (Figs. 1A and
85 1B). Specifically, quantified data showed that the sub-G1 population greatly increased, from 6% at
86 8 h to 40% within a period of 24 h (Fig. 1C). Thus, we examined several cell-cycle regulators: (1)
87 cyclin D1 and its catalytic partner CDK4/6 that control early G1 phase; as well as (2) cyclin E and
88 CDK2 which phosphorylate Rb (Rb phosphorylation is the critical step in cell-cycle progression
9

ACCEPTED MANUSCRIPT
1
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
2
2
89 from G1 to S phase). Exposure to compound 10 led to the upregulation of cyclin D1, which was
90 then sustained at a high level (Fig. 2). In contrast, compound 10 induced a short-term elevation of
91 cyclin E levels, which was followed by a decrease and the attenuation of Rb phosphorylation.
92 Taken together, these data suggest that compound 10 induces cell cycle arrest and apoptosis during
93 early G1 phase.
94
95 2.5. Molecular modeling
96
Treating A549 lung cancer cells with compound 10 was found to arrest cell cycle progression
97 in the early G1 phase by attenuating Rb phosphorylation and the upregulation of cyclin D1. This
98 suggests that CDK2 and CDK4/6 could present molecular targets for compound 10 in the
99 inhibition of cancer cell proliferation. To identify the binding mechanism between compound 10
00 and CDKs, we performed the molecular docking model based on the inhibitor binding mode of
01 CDK4 mimic CDK2 protein structure, as shown in Fig. 3 [18]. Through this investigation, we
02 determined that there are four residues capable of forming hydrogen bonding interactions with
03 compound 10 in the active site of CDK4: Gly 15, Lys 35, Val 96, and Asp 97. The residues Val 96
04 and Asp 97 are found in the front wall of the binding pocket. In addition, the carbonyl oxygen of
05 the oxindole ring in compound 10 likely forms hydrogen bonds with two residues (Gly 15 and Lys
06 35) in the binding pocket, while the methoxy group at 5-position of the oxindole ring may interact
07 with Val 96 through hydrogen bonding. The residue of Phe 93 inside the binding pocket is parallel
08 with the oxindole moiety of compound 10, which may facilitate hydrophobic interactions. Finally,
09 the isoxazolyl ring of compound 10 is buried within the protein cleft, and the 6-quinolinyl ring of
10 this compound protrudes out from the pocket. This structure allows the amido group of
11 6-quinolinyl to form a hydrogen bond with Asp 97.
1
0

ACCEPTED MANUSCRIPT
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
12
13 2.6. 3-Ylideneoxindole acetamide induces caspase-mediated apoptosis in cancer cells
14 Caspase activity assays were performed to determine whether caspases were responsible for
15 apoptosis in cells treated with compound 10. Caspases -3, -8, and -9 were activated in cells treated
16 with compound 10 for 24 h (Fig. 4A). In addition, the caspase-3 substrate PARP was cleaved in a
17 time-dependent manner (Fig. 4B). Collectively, these data suggest that (1) compound 10 inhibits
18 the growth of cancer cells, and (2) the antiproliferative effects of compound 10 are mediated
19 through the induction of apoptosis, such that compound 10-induced apoptosis is
20 caspase-dependent, as shown by the activation of capase-3 in the cleavage of poly(ADP-ribose)
21 polymerase.
22
23 2.7. 3-Ylideneoxindole acetamide inhibited tumor growth in the colon cancer CT-26 xenograft
24 animal model
25
The in vivo activity of 3-ylideneoxindole acetamide was examined in a colon cancer CT-26
26 xenograft animal model using Balb/c mice. Mice treated with cisplatin (2 mg/kg, daily, ip) were
27 used as a positive control to test the response sensitivity of the testing system. Intraperitoneal
28 administration of compound 10 (10 mg/kg) caused a significant inhibition of tumor growth similar
29 to that induced by cisplatin but without loss of body weight (Fig. 5). Conversely, cisplatin causes a
30 loss of body weight in mice. This suggests that compound 10 only had minor toxic effects under
31 these treatment dosages. The animal model further revealed that the in vivo anticancer activity of
32 compound 10 leads to a reduction in the tumor size that is comparable to that of cisplatin, but
33 without the toxicity.
34
1
1

ACCEPTED MANUSCRIPT
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
35 3. Conclusions
36 Considering meisoindigo as a structural template for the development of new drugs, we
37 report the synthesis and biological functions of novel 3-ylideneoxindole acetamides as potent
38 anticancer agents. A series of 3-ylideneoxindole acetamides were synthesized and evaluated for
39 cytotoxic effects against various cancer cell lines. Structure-activity relationships (SARs) revealed
2
40 that the electron deficiency associated with para-phenyl in the R position plays an important role
41 in the cytotoxic activity of 3-ylideneoxindole acetamide. Compound 10, with a methoxy group at
1
3
42 R and a 6-quinolinyl group at R , demonstrated outstanding anticancer activity both in vitro and
43 in vivo. An investigation of the cellular mechanisms behind these antiproliferative effects showed
44 that compound 10 arrested cells in the G1 phase by causing the upregulation of cyclin D1 and the
45 attenuation of RB phosphorylation, which subsequently triggered caspase-dependent apoptosis.
46 Furthermore, a docking model suggests that compound 10 binds to the active site of CDK4
47 through hydrophobic interactions with the residue of Phe 93 as well as hydrogen interactions with
48 four residues (Gly 15, Lys 35, Val 96, and Asp 97). These results suggest that CDKs are potential
49 molecular targets for compound 10 in the inhibition of cancer cell proliferation. Finally, we
50 demonstrated that compound 10 has anticancer activities in a CT26-xenografted BALB/c mouse
51 model when administered via the i.p. route. Specifically, this treatment resulted in reduced tumor
52 size and presented effects comparable to those of cisplatin. However, compound 10 is less toxic
53 than cisplatin; thus, treatment did not cause a loss of body weight. Overall, our in vitro and in vivo
54 investigations revealed that 3-ylideneoxindole acetamides are promising anticancer compounds.
55 However, further studies are required to elucidate the cellular mechanisms involved in the
56 regulation of antiproliferative effects and apoptosis induced by these acetamides.
57
1
2

ACCEPTED MANUSCRIPT
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
58 4. Experimental section
59 4.1. Chemistry synthesis
ο
60
All reactions were conducted in dried glassware under an oven at 120 C overnight and
61 cooled in a desiccator. All reagents were used as received from commercial suppliers unless
62 otherwise stated. Dichloromethane (DCM) and N, N’-dimethylformamide (DMF) were dried over
63 calcium hydride for 48 h prior to distillation. Tetrahydrofuran (THF) was distilled from
64 sodium/benzophenone ketyl under nitrogen. The proton NMR spectra were obtained on Bruker
65 Avance 400 (400 MHz), Varian Unity Inova 500 (500 MHz) and Varian VNMRS600 (600 MHz)
66 spectrometers. Deuterated solvents, CDCl and DMSO-d , of spectrograde were used as solvent.
3
6
67 All NMR chemical shifts were reported as δ values in parts per million (ppm), and coupling
68 constants (J) were given in hertz (Hz). The splitting pattern abbreviations are as follows: s, singlet;
69 d, doublet; t, triplet; q, quartet; br, broad; m, unresolved multiplet due to the field strength of the
70 instrument; dd, doublet of doublet; dt, doublet of triplet; and ddd, doublet of doublet of doublet.
71 Melting points were measured on a Yanaco MP-S3 micro melting point apparatus and are
72 uncorrected. Mass spectra were carried out on ThermoQuest Finnigan and Microsaic 4000MiD
73 mass spectrometers. Purification was performed by using preparative separations in flash column
74 chromatography (Merck silica gel 60, particle size of 230-400 mesh). Analytical TLC was carried
75 out on precoated plates (Merck silica gel 60, F254). Compounds analyzed on the TLC plates were
2 4
76 visualized by using UV light, I vapor, or basic aqueous potassium permanganate (KMnO ) with
77 heating. RPMI 1640 medium, fetal bovine serum (FBS), penicillin, streptomycin, and all other
78 tissue culture regents were obtained from GIBCO/BRL Life Technologies (Grand Island, NY).
79 Antibodies to poly(ADP-ribose)polymerase (PARP), cyclins D1 and E, and α-tubulin were
1
3

ACCEPTED MANUSCRIPT
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
3
3
80 obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA). Antibodies to Rb, and
81 phosphor-Rb Ser780, were obtained from Cell Signaling Technologies (Boston, MA). Caspase
82 activity assay kits for caspases-3, -8, and -9 were obtained from BioVision (Mountain View, CA).
83 Sulfurhodamin B (SRB), cisplatin, and propidium iodide (PI) were obtained from Sigma (St.
84 Louis, MO). MTS and PMS were purchased from Promega Corp. (Madison, WI).
85
86 4.1.1. General procedure for synthesis of (E)-2-(1-(4-chlorobenzyl)-2-oxoindolin-3-ylidene)
87 -N-(pyridin-4-yl)acetamide (4).
88
To a solution of isatin (300 mg, 2.04 mmol) in DMF (10 mL) was added NaH (60%
o
89 dispersion in mineral oil, 86 mg, 2.14 mmol) portionwise at 0 °C. The mixture was stirred at 0 C
90 for 30 min and 4-chlorobenzyl chloride (328 mg, 2.04 mmol) was added dropwise. The reaction
91 mixture was stirred at room temperature for 12 h until complete by TLC. The reaction was
92 quenched by addition of water and extracted with ethyl acetate. The combined organic layers were
93 washed with brine solution, dried over MgSO and concentrated in vacuo. The residue was
4
94 purified by flash column chromatography furnished the desired N-4-chlorobenzyl isatin (466 mg,
95 84%) as an organic solid.
96
2
To a solution of N-4-chlorobenzyl isatin (200 mg, 0.74 mmol) in THF/H O (6/1, 5 mL) was
97 added diethylphosphonoacetamide (200 mg, 0.74mmol) and potassium carbonate (358 mg, 2.6
98 mmol). The mixture was stirred at room temperature for 8 h until complete by TLC, and then
99 poured into iced cold water whereupon a yellowish solid precipitates. The crude mixture was
00 purified by flash column chromatography to afford the title compound 4 (233 mg, 81%) as a
o
1
6
01 yellowish solid. Mp = 208-210 C. H NMR (600 MHz, DMSO-d ) δ: 11.19 (s, 1H), 8.62 (d, J =
1
4

ACCEPTED MANUSCRIPT
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
02 7.2 Hz, 1H), 8.49 (d, J = 6.0 Hz, 2H), 7.69 (dd, J = 4.8, 1.2 Hz, 1H), 7.40-7.38 (m, 3H), 7.35 (d, J
03 = 9.0 Hz, 2H), 7.19 (s, 1H), 7.07 (dt, J = 7.8, 0.6 Hz, 1H), 6.97 (d, J = 7.8 Hz, 1H), 4.97 (s, 2H).
1
3
04
C NMR (150 MHz, DMSO-d ) δ: 167.2, 163.6, 150.6, 145.2, 144.5, 135.3, 135.2, 132.5, 132.1,
6
+
05 129.1, 128.7, 128.5, 126.3, 122.6, 119.5, 113.4, 109.5, 42.2. HRMS calcd for C H N O Cl (M)
2
2
16
3
2
06 389.0911, found 389.0921.
07
08 4.1.1.1. (E)-2-(1-(4-Chlorobenzyl)-2-oxoindolin-3-ylidene)-N-(quinolin-6-yl)acetamide (5). Yield:
o
1
09 75%. Mp = 248-250 C. H NMR (600 MHz, DMSO-d ) δ: 11.20 (s, 1H), 8.80 (dd, J = 4.2, 1.8 Hz,
6
10 1H), 8.74 (d, J = 2.4, 1H), 8.64 (d, J = 2.4 Hz, 1H), 8.37 (d, J = 7.8 Hz, 1H), 8.02 (d, J = 9.0 Hz,
11 1H), 7.87 (dd, J = 9.0, 2.4 Hz, 1H), 7.51 (dd, J = 8.4, 4.2 Hz, 1H), 7.40 (d, J = 8.4 Hz, 2H), 7.36 (d,
12 J = 8.4 Hz, 2H), 7.34 (m, 1H), 7.29 (s, 1H), 7.08 (t, J = 7.8 Hz, 1H), 6.97 (d, J = 8.4 Hz, 1H), 4.98
1
3
13 (s, 2H). C NMR (150 MHz, DMSO-d ) δ: 167.3, 162.9, 149.4, 145.0, 144.4, 136.6, 135.7, 135.2,
6
14 134.7, 132.2, 132.1, 129.8, 129.1, 128.7, 128.6, 128.3, 127.1, 123.2, 122.6, 121.9, 119.7, 115.5,
+
15 109.4, 42.2. HRMS calcd for C26
H
18
N
3
O
2
Cl (M) 439.1078, found 439.1083.
16
17 4.1.1.2.
18 (E)-N-(4-Methoxyphenyl)-2-(1-((3-methylisoxazol-5-yl)methyl)-2-oxoindolin-3-ylidene)acetamide
o
1
19 (6). Yield: 74%. Mp = 208-210 C. H NMR (600 MHz, DMSO-d ) δ: 10.76 (s, 1H), 8.70 (d, J =
6
20 7.8 Hz, 1H), 7.68 (dd, J = 7.2, 2.4 Hz, 2H), 7.41-7.38 (m, 1H), 7.18 (s, 1H), 7.11-7.08 (m, 2H),
1
3
21 6.94 (dd, J = 7.2, 2.4 Hz, 2H), 6.35 (s, 1H), 5.11 (s, 2H), 3.74 (s, 3H), 2.17 (s, 3H). C NMR (150
22 MHz, DMSO-d ) δ: 167.0, 166.5, 161.8, 159.8, 155.8, 143.7, 133.6, 132.0, 132.0, 128.6, 127.9,
6
+
23 122.7, 120.8, 119.7, 114.0, 109.2, 103.9, 55.2, 35.2, 10.9. HRMS calcd for C H N O (M)
2
2
19
3
4
1
5

ACCEPTED MANUSCRIPT
324 389.1396, found 389.1386.
325
326 4.1.1.3.
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
27 (E)-2-(1-((3-Methylisoxazol-5-yl)methyl)-2-oxoindolin-3-ylidene)-N-(pyridin-4-yl)acetamide (7).
o
1
28 Yiled: 80%. Mp = 210-214 C. H NMR (600 MHz, DMSO-d ) δ: 11.18 (s, 1H), 8.64 (d, J = 7.2
6
29 Hz, 1H), 8.49 (d, J = 6.0 Hz, 2H), 7.68 (dd, J = 4.8, 1.2 Hz, 2H), 7.43 (dt, J = 7.8, 1.2 Hz, 1H),
1
3
30 7.17 (s, 1H), 7.11 (d, J = 7.8 Hz, 2H), 6.36 (s, 1H), 5.11 (s, 2H), 2.17 (s, 3H). C NMR (150 MHz,
31 DMSO-d ) δ: 166.8, 166.4, 163.5, 159.8, 150.6, 145.2, 144.1, 135.0, 132.5, 128.5, 126.4, 122.8,
6
+
32 119.4, 113.4, 109.4, 103.9, 35.2, 10.9. HRMS calcd for C H N O (M) 360.1245, found
2
2
19
3
4
33 360.1234.
34
35 4.1.1.4.
36 (E)-2-(1-((3-Methylisoxazol-5-yl)methyl)-2-oxoindolin-3-ylidene)-N-(quinolin-6-yl)acetamide (8).
o
1
37 Yield: 78%. Mp = 250-252 C. H NMR (600 MHz, DMSO-d ) δ: 11.20 (s, 1H), 8.82-8.80 (m,
6
38 1H), 8.75 (dd, J = 8.4, 1.2 Hz, 1H), 8.63 (d, J = 2.4 Hz, 1H), 8.37 (dd, J = 8.4, 1.2 Hz, 1H), 8.01 (d,
39 J = 9.0 Hz, 1H), 7.86 (dd, J = 9.0, 2.4 Hz, 1H), 7.51 (dd, J = 8.4, 4.2 Hz, 1H), 7.43 (dt, J = 7.8, 1.2
1
3
40 Hz, 1H), 7.27 (s, 1H), 7.13 (t, J = 7.8 Hz, 2H), 6.37 (s, 1H), 5.14 (s, 2H), 2.18 (s, 3H). C NMR
41 (150 MHz, DMSO-d ) δ: 167.0, 166.5, 162.8, 159.8, 149.4, 145.0, 143.9, 136.6, 135.7, 134.4,
6
42 132.3, 129.8, 128.6, 128.3, 127.3, 123.2, 122.8, 121.9, 119.6, 115.5, 109.3, 103.9, 35.2, 10.9.
+
43 HRMS calcd for C24
H
18
N
4
O
3
(M) 410.1359, found 410.1369.
44
45 4.1.1.5.
46 (E)-N-(4-Hydroxyphenyl)-2-(1-((3-methylisoxazol-5-yl)methyl)-2-oxoindolin-3-ylidene)acetamide
1
6

ACCEPTED MANUSCRIPT
o
1
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
47 (9). Yield: 74%. Mp = 255-257 C. H NMR (600 MHz, DMSO-d
6
) δ: 10.66 (s, 1H), 9.35 (s, 1H),
48 8.70 (d, J = 7.8 Hz, 1H), 7.56 (d, J = 9.0 Hz, 2H), 7.39 (t, J = 7.8 Hz, 1H), 7.17 (s, 1H), 7.10-7.07
1
3
49 (m, 2H), 6.75 (d, J = 8.4 Hz, 2H), 6.34 (s, 1H), 5.11 (s, 2H), 2.17 (s, 3H). C NMR (150 MHz,
50 DMSO-d
6
) δ: 167.0, 166.5, 161.6, 159.8, 154.1, 143.7, 133.4, 131.8, 130.5, 128.6, 128.2, 122.6,
+
51 121.0, 119.8, 115.3, 109.2, 103.9, 35.2, 10.9. HRMS calcd for C H N O (M) 375.1239, found
2
1
17
3
4
52 375.1229.
53
54 4.1.1.6.
55 (E)-2-(5-Methoxy-1-((3-methylisoxazol-5-yl)methyl)-2-oxoindolin-3-ylidene)-N-(quinolin-6-yl)ace
o
1
6
56 tamide (10). Yield: 78%. Mp = 192-195 C. H NMR (600 MHz, DMSO-d ) δ: 11.20 (s, 1H), 8.81
57 (dd, J = 4.2, 1.8 Hz, 1H), 8.61 (d, J = 1.8 Hz, 1H), 8.45 (s, 1H), 8.38 (d, J = 7.8 Hz, 1H), 8.01 (d, J
58 = 9.0 Hz, 1H), 7.88 (dd, J = 9.0, 2.4 Hz, 1H), 7.50 (dd, J = 8.4, 4.2 Hz, 1H), 7.26 (s, 1H), 7.03 (s,
1
3
6
59 1H), 6.35 (s, 1H), 5.10 (s, 2H), 3.77 (s, 3H), 2.17 (s, 3H). C NMR (150 MHz, DMSO-d ) δ:
60 166.8, 166.6, 162.8, 159.8, 155.3, 149.4, 145.1, 137.6, 136.6, 135.8, 134.9, 129.8, 128.3, 127.6,
61 123.3, 121.9, 120.5, 116.7, 115.7, 115.6, 109.7, 103.9, 55.7, 35.3, 10.9. HRMS calcd for
+
62 C H N O (M+H) 441.1563, found 441.1576.
25
21
4
4
63
64 4.1.1.7.
65 (E)-N-(2-Iodophenyl)-2-(1-((3-methylisoxazol-5-yl)methyl)-2-oxoindolin-3-ylidene)acetamide (11).
o
1
66 Yield: 84%. Mp = 206-207 C. H NMR (600 MHz, DMSO-d ) δ: 10.41 (s, 1H), 8.61 (d, J = 7.8
6
67 Hz, 1H), 7.92 (d, J = 7.8 Hz, 1H), 7.53 (d, J = 7.8 Hz, 1H), 7.44 (t, J = 7.8 Hz, 1H), 7.40 (t, J = 7.8
68 Hz, 1H), 7.30 (s, 1H), 7.10 (d, J = 7.8 Hz, 1H), 7.07-7.03 (m, 2H), 6.36 (s, 1H), 5.12 (s, 2H), 2.17
17

ACCEPTED MANUSCRIPT
1
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
6
69 (s, 3H). C NMR (150 MHz, DMSO-d ) δ: 166.9, 166.5, 162.7, 159.8, 143.9, 139.0, 139.0, 134.5,
70 132.2, 128.7, 128.6, 128.3, 127.6, 127.1, 122.6, 119.6, 109.2, 103.9, 96.7, 35.2, 10.9. HRMS calcd
+
71 for C H IN O (M) 485.0214, found 485.0225.
2
1
16
3
3
72
73 4.1.1.8.
74 (E)-N-(3-Iodophenyl)-2-(1-((3-methylisoxazol-5-yl)methyl)-2-oxoindolin-3-ylidene)acetamide (12).
o
1
75 Yield: 77%. Mp = 187-189 C. H NMR (600 MHz, DMSO-d ) δ: 10.94 (s, 1H), 8.65 (dd, J = 7.8,
6
76 0.6 Hz, 1H), 8.29 (t, J = 1.8 Hz, 1H), 7.63 (dd, J = 7.8, 1.2 Hz, 1H), 7.48 (d, J = 7.8 Hz, 1H), 7.42
77 (dt, J = 7.8, 1.2 Hz, 1H), 7.18-7.15 (m, 3H), 7.12-7.10 (m, 2H), 6.35 (s, 2H), 5.11 (s, 2H), 2.17 (s,
1
3
6
78 3H). C NMR (150 MHz, DMSO-d ) δ: 167.0, 166.5, 162.6, 160.0, 143.9, 140.1, 134.5, 132.7,
79 132.3, 131.0, 128.6, 127.5, 127.1, 122.8, 119.6, 118.7, 109.3, 103.9, 94.8, 35.2, 10.9. HRMS calcd
+
80 for C H IN O (M) 485.0243, found 485.0240.
2
1
16
3
3
81
82 4.1.1.9. (E)-2-(1-((3-Methylisoxazol-5-yl)methyl)-2-oxoindolin-3-ylidene)-N-phenylacetamide (13).
o
1
83 Yield: 84%. Mp = 201-204 C. H NMR (600 MHz, DMSO-d ) δ: 10.86 (s, 1H), 8.68 (d, J = 7.8
6
84 Hz, 1H), 7.75 (d, J = 8.4 Hz, 2H), 7.41 (t, J = 7.2 Hz, 1H), 7.36 (t, J = 7.8 Hz, 2H), 7.21 (s, 1H),
1
3
6
85 7.14-7.08 (m, 3H), 6.35 (s, 1H), 5.12 (s, 2H), 2.17 (s, 3H). C NMR (150 MHz, DMSO-d ) δ:
86 167.0, 166.5, 162.4, 159.8, 143.8, 138.7, 134.0, 132.1, 128.9, 128.5, 127.7, 124.2, 122.7, 119.6,
+
87 119.4, 109.2, 103.9, 35.2, 10.9. HRMS calcd for C H N O (M) 359.1252, found 359.1261.
2
1
17
3
3
88
89 4.1.1.10.
90 (E)-2-(1-((3-Methylisoxazol-5-yl)methyl)-2-oxoindolin-3-ylidene)-N-(4-nitrophenyl)acetamide
1
8

ACCEPTED MANUSCRIPT
o
1
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
4
4
4
6
91 (14). Yield: 76%. Mp = 210-214 C. H NMR (600 MHz, DMSO-d ) δ: 11.41 (s, 1H), 8.66 (d, J =
92 7.8 Hz, 1H), 8.27 (d, J = 9.0 Hz, 1H), 7.99 (d, J = 9.0 Hz, 2H), 7.43 (t, J = 7.8 Hz, 1H), 7.20 (d, J
93 = 8.4 Hz, 1H), 7.20 (s, 1H), 7.13-7.10 (m, 2H), 6.86 (d, J = 8.4 Hz, 1H), 6.36 (s, 1H), 5.12 (s, 2H),
1
3
6
94 2.17 (s, 3H). C NMR (150 MHz, DMSO-d ) δ: 166.8, 166.4, 163.2, 159.8, 144.7, 144.1, 142.8,
95 135.1, 132.6, 128.6, 126.4, 125.1, 122.8, 119.3, 113.7, 109.4, 103.9, 35.2, 10.8. HRMS calcd for
+
96 C H N O (M) 404.1101, found 404.1111.
21
16
4
5
97
98 4.1.1.11. (E)-Methyl 4-((2-oxo-3-(2-oxo-2-(quinolin-6-ylamino)ethylidene)indolin-1-yl)methyl)
o
1
3
99 benzoate (15). Yield: 78%. Mp = 147-150 C. H NMR (600 MHz, CDCl ) δ: 8.56 (dd, J = 4.2,
00 1.2 Hz, 1H), 8.77 (d, J = 7.8 Hz, 1H), 8.52 (d, J = 2.4 Hz, 1H), 8.15 (d, J = 7.2 Hz, 2H), 8.06 (d, J
01 = 9.0 Hz, 1H), 7.98 (d, J = 7.8 Hz, 2H), 7.65 (dd, J = 9.0, 2.4 Hz, 1H), 7.40 (dd, J = 8.4, 4.2 Hz,
02 1H), 7.26 (d, J = 7.2 Hz, 1H), 7.15 (s, 1H), 7.08 (t, J = 7.8 Hz, 1H), 6.63 (d, J = 7.8 Hz, 1H), 5.01
13
3
03 (s, 2H), 3.88 (s, 3H). C NMR (150 MHz, CDCl ) δ: 168.1, 166.6, 162.7, 149.8, 145.9, 144.5,
04 140.4, 136.5, 135.9, 135.4, 132.4, 130.6, 130.2, 129.8, 129.5, 128.8, 127.0, 125.7, 123.4, 123.1,
+
05 121.8, 120.2, 116.6, 109.0, 52.2, 43.7. ESMS m/z: 462.4 (M‒1) .
06
07 4.1.1.12. (E)-2-(1-(Naphthalen-2-ylmethyl)-2-oxoindolin-3-ylidene)-N-(quinolin-6-yl)acetamide
o
1
3
08 (16). Yield: 79%. Mp = 231-233 C. H NMR (600 MHz, CDCl ) δ: 8.83 (d, J = 3.0 Hz, 1H), 8.78
09 (d, J = 7.2 Hz, 1H), 8.50 (d, J = 1.8 Hz, 1H), 8.48 (s, 1H), 8.09 (d, J = 7.8 Hz, 1H), 8.02 (d, J = 9.6
10 Hz, 1H), 7.81-7.52 (m, 3H), 7.69 (s, 1H), 7.68 (dd, J = 9.6, 2.4 Hz, 1H), 7.46-7.44 (m, 2H), 7.40
11 (dd, J = 8.4, 1.8 Hz, 1H), 7.36 (dd, J = 8.4, 4.2 Hz, 1H), 7.26-7.21 (m, 2H), 7.06 (t, J = 7.8 Hz,.
1
3
3
12 1H), 6.72 (d, J = 7.8 Hz, 1H), 5.13 (s, 2H). C NMR (150 MHz, CDCl ) δ: 168.4, 162.8, 149.7,
1
9

ACCEPTED MANUSCRIPT
4
4
4
4
4
4
4
4
4
4
4
4
4
13 145.8, 144.8, 136.6, 136.0, 135.6, 133.3, 132.9, 132.6, 132.3, 129.4, 128.9, 128.8, 127.7, 126.5,
14 126.2, 125.9, 125.8, 124.9, 123.3, 123.1, 121.8, 120.2, 116.6, 109.3, 44.2. HRMS calcd for
+
15 C H N O (M+H) 456.1718, found 456.1720.
30
21
3
2
16
17 4.1.1.13. (E)-2-(2-Oxo-1-(4-(trifluoromethyl)benzyl)indolin-3-ylidene)-N-(quinolin-6-yl)acetamide
o
1
6
18 (17). Yield: 83%. Mp = 231-233 C. H NMR (600 MHz, DMSO-d ) δ: 11.21 (s, 1H), 8.81 (dd, J
19 = 4.2, 1.8 Hz, 1H), 8.76 (d, J = 7.2 Hz, 1H), 8.65 (d, J = 1.8 Hz, 1H), 8.37 (d, J = 7.8 Hz, 1H),
20 8.02 (d, J = 9.0 Hz, 1H), 7.87 (dd, J = 9.0, 2.4 Hz, 1H), 7.71 (d, J = 8.4 Hz, 2H), 7.55 (d, J = 8.4
21 Hz, 2H), 7.51 (dd, J = 8.4, 4.2 Hz, 1H), 7.36 (dt, J = 7.8, 1.2 Hz, 1H), 7.30 (s, 1H), 7.10 (s, 1H),
13
22 6.97 (d, J = 7.8 Hz, 1H), 5.10 (s, 2H). C NMR (150 MHz, DMSO-d ) δ: 167.4, 162.9, 149.4,
6
23 145.0, 144.3, 141.1, 136.6, 135.7, 134.7, 132.2, 129.8, 128.7, 128.3, 128.0, 127.2, 125.6 (t, J =
24 3.46 Hz), 125.0, 123.2, 122.6, 121.9, 119.7, 115.5, 109.4, 42.4. HRMS calcd for C H N O F
2 3
2
7
19
3
+
25 (M+H) 474.1409, found 474.1405.
426
427 4.1.1.14. (E)-2-(1-(4-Bromobenzyl)-2-oxoindolin-3-ylidene)-N-(quinolin-6-yl)acetamide (18).
o
1
4
4
4
4
4
4
4
4
3
28 Yield: 78%. Mp = 228-232 C. H NMR (600 MHz, CDCl ) δ: 8.84 (dd, J = 3.6, 1.2 Hz, 1H), 8.79
29 (d, J = 7.8 Hz, 1H), 8.63 (s, 1H), 8.51 (d, J = 1.8 Hz, 1H), 8.11 (d, J = 8.4 Hz, 1H), 8.01 (d, J = 9.6
30 Hz, 1H), 7.66 (dd, J = 2.4, 1.8 Hz, 1H), 7.42 (d, J = 7.8 Hz, 2H), 7.39 (dd, J = 8.4, 4.2 Hz, 1H),
31 7.26 (t, J = 7.8 Hz, 1H), 7.24 (s, 1H), 7.14 (d, J = 8.4 Hz, 2H), 7.08 (t, J = 7.8 Hz, 1H), 6.64 (d, J
1
3
3
32 = 7.8 Hz, 1H), 4.91 (s, 2H). C NMR (150 MHz, CDCl ) δ: 168.3, 162.8, 149.7, 145.8, 144.4,
33 136.3, 136.0, 135.6, 134.2, 132.3, 132.1, 130.4, 129.6, 128.8, 128.7, 126.2, 123.4, 123.2, 121.8,
+
34 120.2, 116.6, 109.0, 43.4. HRMS calcd for C H N O Br (M+H) 484.0660, found 484.0657.
2
6
19
3
2
35
2
0

ACCEPTED MANUSCRIPT
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
36 4.1.1.15. (E)-2-(1-(3-Cyanobenzyl)-2-oxoindolin-3-ylidene)-N-(quinolin-6-yl)acetamide (19).
o
1
37 Yield: 77%. Mp = 192-194 C. H NMR (600 MHz, DMSO-d ) δ: 11.20 (s, 1H), 8.80 (d, J = 3.0
6
38 Hz, 1H), 8.76 (d, J = 7.8 Hz, 1H), 8.64 (s, 1H), 8.36 (d, J = 8.4 Hz, 1H), 8.01 (d, J = 9.0 Hz, 1H),
39 7.88-7.86 (m, 2H), 7.75 (d, J = 7.5 Hz, 1H), 7.65 (d, J = 7.8 Hz, 1H), 7.55 (t, J = 7.8 Hz, 1H), 7.50
40 (dd, J = 8.4, 4.2 Hz, 1H), 7.36 (t, J = 7.8 Hz, 1H), 7.29 (s, 1H), 7.10 (t, J = 7.8 Hz, 1H), 6.99 (d, J
1
3
41 = 7.8 Hz, 1H), 5.04 (s, 2H). C NMR (150 MHz, DMSO-d ) δ: 167.5, 162.9, 149.4, 145.0, 144.3,
6
42 138.0, 136.7, 135.7, 134.8, 132.2, 132.0, 131.4, 130.9, 130.0, 129.8, 128.7, 128.3, 127.1, 123.3,
+
43 122.6, 121.9, 119.8, 118.6, 115.5, 111.6, 109.3, 42.2. HRMS calcd for C H N O (M+H)
2
7
19
4
2
44 431.1508, found 431.1513.
45
46 4.1.1.16. (E)-2-(5-Bromo-1-(4-chlorobenzyl)-2-oxoindolin-3-ylidene)-N-(quinolin-6-yl)acetamide
o
1
47 (20). Yield: 79%. Mp = 256-258 C. H NMR (600 MHz, DMSO-d ) δ: 11.28 (s, 1H), 8.98 (d, J =
6
48 2.4 Hz, 1H), 8.82 (dd, J = 4.2, 1.2 Hz, 1H), 8.63 (d, J = 1.8 Hz, 1H), 8.44 (d, J = 9.0 Hz, 1H), 8.02
49 (d, J = 9.0 Hz, 1H), 7.87 (dd, J = 9.0, 2.4 Hz, 1H), 7.57 (dd, J = 7.8, 2.4 Hz, 1H), 7.51 (dd, J = 7.8,
50 4.2 Hz, 1H), 7.40 (d, J = 8.4 Hz, 2H), 7.35 (d, J = 8.4 Hz, 2H), 6.95 (d, J = 7.8 Hz, 1H), 4.99 (s,
1
3
51 2H). C NMR (150 MHz, DMSO-d ) δ: 166.9, 162.6, 149.5, 145.1, 143.6, 136.5, 135.9, 134.9,
6
52 134.4, 133.8, 132.2, 130.9, 129.8, 129.1, 128.8, 128.7, 128.3, 123.3, 121.9, 121.7, 115.7, 114.3,
+
53 111.4, 42.3. HRMS calcd for C H N O ClBr (M+H) 518.0270, found 518.0273.
2
6
18
3
2
54
55 4.1.1.17. (E)-2-(1-(4-Cyanobenzyl)-2-oxoindolin-3-ylidene)-N-(quinolin-6-yl)acetamide (21).
o
1
56 Yield: 78%. Mp = 237-240 C. H NMR (600 MHz, DMSO-d ) δ: 11.21 (s, 1H), 8.81 (dd, J = 3.6,
6
57 1.2 Hz, 1H), 8.76 (d, J = 7.8 Hz, 1H), 8.64 (d, J = 1.8 Hz, 1H), 8.38 (d, J = 8.4 Hz, 1H), 8.01 (d, J
2
1

ACCEPTED MANUSCRIPT
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
58 = 9.0 Hz, 1H), 7.88-7.86 (m, 2H), 7.76 (d, J = 7.2 Hz, 1H), 7.65 (d, J = 7.8 Hz, 1H), 7.56 (t, J =
59 7.8 Hz, 1H), 7.51 (dd, J = 8.4, 3.6 Hz, 1H), 7.37 (t, J = 7.8 Hz, 1H), 7.29 (s, 1H), 7.10 (t, J = 7.8
1
3
60 Hz, 1H), 6.99 (d, J = 8.4 Hz, 1H), 5.05 (s, 2H). C NMR (150 MHz, DMSO-d ) δ: 167.9, 163.4,
6
61 149.8, 145.5, 144.7, 138.4, 137.1, 136.2, 135.2, 132.6, 132.5, 131.8, 130.5, 130.2, 129.1, 128.8,
19 4 2
62 127.6, 123.7, 123.1, 122.4, 120.3, 119.0, 115.9, 112.1, 109.8, 42.6. HRMS calcd for C27H N O
+
63 (M+H) 431.1508, found 431.1504.
64
65 4.1.1.18. (E)-2-(1-(4-Nitrobenzyl)-2-oxoindolin-3-ylidene)-N-(quinolin-6-yl)acetamide (22). Yield:
o
1
66 76%. Mp = 272-275 C. H NMR (600 MHz, DMSO-d ) δ: 11.22 (s, 1H), 8.81 (dd, J = 4.2, 1.8 Hz,
6
67 1H), 8.77 (d, J = 7.8 Hz, 1H), 8.65 (d, J = 1.8 Hz, 1H), 8.38 (dd, J = 7.8, 1.2 Hz, 1H), 8.21 (dt, J =
68 9.0, 1.8 Hz, 2H), 8.02 (d, J = 9.6 Hz, 1H), 7.87 (dd, J = 9.0, 1.8 Hz, 1H), 7.60 (d, J = 9.0 Hz, 2H),
69 7.51 (dd, J = 8.4, 4.2 Hz, 1H), 7.37 (dt, J = 7.2, 1.2 Hz, 1H), 7.31 (s, 1H), 7.11 (dt, J = 7.8, 1.2 Hz,
1
3
70 1H), 6.97 (d, J = 7.8 Hz, 1H), 5.15 (s, 2H). C NMR (150 MHz, DMSO-d ) δ: 167.4, 162.8, 149.4,
6
71 146.9, 145.0, 144.2, 144.1, 136.6, 135.8, 134.6, 132.2, 129.8, 128.7, 128.3, 128.3, 127.3, 123.9,
+
72 123.2, 122.7, 121.9, 119.8, 115.5, 109.4, 42.4. HRMS calcd for C26
H N
19 4
O
4
(M+H) 451.1396,
73 found 451.1393.
74
75 4.1.1.19. (E)-2-(1-(3-Chlorobenzyl)-2-oxoindolin-3-ylidene)-N-(quinolin-6-yl)acetamide (23)
o
1
76 Yield: 78%. Mp = 242-245 C. H NMR (600 MHz, CDCl ) δ: 11.20 (s, 1H), 8.81 (dd, J = 4.2, 1.8
3
77 Hz, 1H), 8.75 (d, J = 7.8 Hz, 1H), 8.64 (d, J = 2.4 Hz, 1H), 8.38 (d, J = 8.4 Hz, 1H), 8.01 (d, J =
78 8.4 Hz, 1H), 7.87 (dd, J = 9.0, 2.4 Hz, 1H), 7.50 (dd, J = 8.4, 4.2 Hz, 1H), 7.43 (s, 1H), 7.38-7.33
1
3
79 (m, 3H), 7.29-7.28 (m, 2H), 7.10 (t, J = 7.8 Hz, 1H), 7.00 (d, J = 7.8 Hz, 1H), 5.00 (s, 2H). C
2
2

ACCEPTED MANUSCRIPT
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
5
5
80 NMR (150 MHz, CDCl
3
) δ: 167.4, 162.9, 149.4, 145.0, 144.3, 138.8, 136.6, 135.8, 134.7, 133.3,
81 132.2, 130.7, 129.8, 128.6, 128.3, 127.5, 127.2, 127.1, 125.8, 123.3, 122.6, 121.9, 119.7, 115.5,
+
82 109.4, 42.3. HRMS calcd for C H N O Cl (M+H) 440.1166, found 440.1173.
2
6
19
3
2
83
84 4.1.1.20. (E)-2-(1-(4-Chlorobenzyl)-5-methyl-2-oxoindolin-3-ylidene)-N-(quinolin-6-yl)acetamide
1
85 (24). Yield: 81%. Amorphous powder. H NMR (600 MHz, DMSO-d ) δ: 11.18 (s, 1H), 8.80 (dd,
6
86 J = 4.2, 1.8 Hz, 1H), 8.61 (d, J = 2.4 Hz, 1H), 8.58 (s, 1H), 8.39 (d, J = 7.8 Hz, 1H), 8.01 (d, J =
87 9.0 Hz, 1H), 7.88 (dd, J = 9.0, 2.4 Hz, 1H), 7.51 (dd, J = 8.4, 4.2 Hz, 1H), 7.39 (dd, J = 6.6, 1.8
88 Hz, 2H), 7.34 (d, J = 8.4 Hz, 2H), 7.26 (s, 1H), 7.16 (d, J = 7.2 Hz, 1H), 7.08 (d, J = 9.0 Hz, 1H),
1
3
89 6.85 (d, J = 8.4 Hz, 1H), 6.82 (d, J = 9.0 Hz, 1H), 4.95 (s, 2H), 2.30 (s, 3H). C NMR (150 MHz,
90 DMSO-d ) δ: 167.4, 162.9, 149.4, 145.0, 142.2, 136.6, 135.8, 135.3, 135.0, 132.5, 132.1, 131.5,
6
91 129.8, 129.1, 129.1, 128.7, 128.3, 127.4, 127.0, 123.3, 122.0, 119.8, 115.5, 113.9, 109.2, 68.8,
+
92 42.2. HRMS calcd for C H N O Cl (M+H) 454.1322, found 454.1321.
2
7
21
3
2
93
94 4.1.1.21. (E)-2-(1-(4-Fluorobenzyl)-2-oxoindolin-3-ylidene)-N-(quinolin-6-yl)acetamide (25).
o
1
95 Yield: 74%. Mp = 221-225 C. H NMR (600 MHz, DMSO-d ) δ: 11.20 (s, 1H), 8.81 (dd, J = 4.2,
6
96 1.8 Hz, 1H), 8.74 (d, J = 7.2 Hz, 1H), 8.64 (d, J = 1.8 Hz, 1H), 8.37 (d, J = 7.8 Hz, 1H), 8.01 (d, J
97 = 8.4 Hz, 1H), 7.87 (dd, J = 9.6, 3.0 Hz, 1H), 7.51 (dd, J = 7.8, 3.6 Hz, 1H), 7.40 (dd, J = 7.8, 1.2
98 Hz, 1H), 7.36 (dt, J = 7.8, 1.2 Hz, 1H), 7.29 (s, 1H), 7.17 (dt, J = 6.6, 1.8 Hz, 2H), 7.10 (dt, J = 7.8,
1
3
99 1.2 Hz, 1H), 7.00 (d, J = 7.8 Hz, 1H), 4.97 (s, 2H). C NMR (150 MHz, DMSO-d ) δ: 167.3,
6
00 162.9, 162.3, 160.7, 149.4, 145.0, 144.4, 136.6, 135.7, 134.8, 132.4, 132.4, 132.2, 129.8, 129.4,
01 129.3, 128.6, 128.3, 127.1, 123.2, 122.5, 121.9, 119.7, 115.6, 115.5, 115.4, 109.5, 42.1. HRMS
2
3

ACCEPTED MANUSCRIPT
+
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
02 calcd for C26
H
19
N
3
O
2
F(M+H) 424.1461, found 424.1460.
03
04 4.1.1.22. (E)-2-(1-(2,6-Dichlorobenzyl)-2-oxoindolin-3-ylidene)-N-(quinolin-6-yl)acetamide (26).
o
1
05 Yield: 77%. Mp = 279-281 C. H NMR (600 MHz, DMSO-d ) δ: 11.17 (s, 1H), 8.81 (d, J = 3.0
6
06 Hz, 1H), 8.72 (d, J = 7.8 Hz, 1H), 8.62 (s, 1H), 8.37 (d, J = 9.0 Hz, 1H), 8.00 (d, J = 8.4 Hz, 1H),
07 7.85 (d, J = 9.0 Hz, 1H), 7.53-7.50 (m, 2H), 7.40 (t, J = 8.4 Hz, 1H), 7.32 (t, J = 8.4 Hz, 1H), 7.23
13
08 (s, 1H), 7.06 (t, J = 7.8 Hz, 1H), 6.73 (d, J =7.8 Hz, 1H), 5.22 (s, 2H). C NMR (150 MHz,
09 DMSO-d ) δ: 166.9, 162.9, 149.4, 145.0, 144.4, 136.6, 135.7, 135.3, 134.4, 132.1, 130.7, 130.2,
6
10 129.8, 129.2, 128.5, 128.3, 127.0, 123.3, 122.3, 121.9, 119.7, 115.5, 109.0, 40.0. HRMS calcd for
+
11
12
C
26
H
18
N
3
O
2
Cl
2
(M+H) 474.0776, found 474.0778.
13 4.1.1.23.
14 (E)-2-(1-(4-Chlorobenzyl)-5-methoxy-2-oxoindolin-3-ylidene)-N-(quinolin-6-yl)acetamide
(27).
o
1
15 Yield: 81%. Mp = 239-241 C. H NMR (600 MHz, CDCl ) δ: 8.85 (dd, J = 4.2, 1.8 Hz, 1H), 8.51
3
16 (dd, J = 4.8, 2.4 Hz, 1H), 8.40 (s, 1H), 8.12 (d, J = 9.0 Hz, 1H), 8.03 (d, J = 8.4 Hz, 1H), 7.67 (dd,
17 J = 9.0, 2.4 Hz, 1H), 7.40 (dd, J = 7.8, 2.4 Hz, 1H), 7.28 (d, J = 6.6 Hz, 2H), 7.22 (d, J = 6.6 Hz,
18 2H), 7.20 (s, 1H), 6.82 (dd, J =8.4, 2.4 Hz, 1H), 6.53 (d, J = 8.4 Hz, 1H), 4.90 (s, 2H), 3.82 (s, 3H).
1
3
19
C NMR (150 MHz, CDCl ) δ: 168.1, 162.6, 156.2, 149.8, 145.9, 138.2, 136.9, 136.0, 135.5,
3
20 133.8, 133.7, 130.5, 129.2, 129.1, 128.8, 128.7, 128.4, 126.2, 123.1, 121.8, 120.9, 118.0, 116.6,
+
21 115.5, 109.5, 56.0, 43.4. HRMS calcd for C27
H N
21 3
O
3
Cl (M+H) 470.1271, found 470.1276.
22
23 4.1.1.24. (E)-2-(1-(4-Methoxybenzyl)-2-oxoindolin-3-ylidene)-N-(quinolin-6-yl)acetamide (28).
2
4

ACCEPTED MANUSCRIPT
o
1
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
24 Yield: 76%. Mp = 216-218 C. H NMR (600 MHz, CDCl
3
) δ: 8.83 (dd, J = 4.2, 1.2 Hz, 1H), 8.69
25 (s, 1H), 8.51 (d, J = 2.4 Hz, 1H), 8.08 (d, J = 8.4 Hz, 1H), 7.99 (d, J = 9.0 Hz, 1H), 7.68 (dd, J =
26 9.0, 2.4 Hz, 1H), 7.38 (dd, J = 8.4, 4.8 Hz, 1H), 7.28-7.21 (m, 5H), 7.06 (t, J = 7.8 Hz, 1H), 6.83
1
3
27 (d, J = 6.6 Hz, 2H), 6.71 (d, J = 7.8 Hz, 1H), 4.91 (s, 2H), 3.76 (s, 3H). C NMR (150 MHz,
28 CDCl ) δ: 168.3, 162.9, 159.2, 149.7, 145.8, 144.7, 136.6, 135.9, 135.7, 132.2, 130.4, 129.4, 128.8,
3
29 128.4, 127.1, 126.0, 123.2, 121.7, 120.2, 116.6, 114.3, 109.3, 55.3, 43.5. HRMS calcd for
+
30 C H N O (M+H) 436.1661, found 436.1661.
27
22
3
3
31
32 4.1.1.25.
33 (E)-N-(3-Methylisoxazol-5-yl)-2-(1-((3-methylisoxazol-5-yl)methyl)-2-oxoindolin-3-ylidene)aceta
o
1
34 mide (29). Yield: 81%. Mp = 253-255 C. H NMR (600 MHz, DMSO-d ) δ: 12.35 (s, 1H), 8.64
6
35 (d, J = 7.8 Hz, 1H), 7.44 (t, J = 7.8 Hz, 1H), 7.14 (s, 1H), 7.11 (t, J = 8.4, 2H), 6.38 (s, 1H), 6.36 (s,
1
3
6
36 1H), 5.10 (s, 2H), 2.21 (s, 3H), 2.17 (s, 3H). C NMR (150 MHz, DMSO-d ) δ: 166.6, 166.4,
37 160.9, 160.6, 160.6, 160.0, 144.2, 136.0, 132.8, 128.5, 124.6, 122.8, 119.4, 109.4, 103.9, 89.8,
+
38 35.2, 11.4, 10.9. HRMS calcd for C H N O (M) 364.1171, found 364.1179.
1
9
16
4
4
39
40 4.1.1.26.
41 (E)-2-(1-((3-Methylisoxazol-5-yl)methyl)-2-oxoindolin-3-ylidene)-N-(pyridin-3-yl)acetamide (30).
o
1
42 Yield: 83%. Mp = 195-197 C. H NMR (600 MHz, DMSO-d
6
) δ: 11.07 (s, 1H), 8.86 (d, J = 2.4
43 Hz, 1H), 8.66 (d, J = 7.8 Hz, 1H), 8.33 (dd, J = 4.2, 1.2 Hz, 1H), 8.21-8.18 (m, 1H), 7.43-7.40 (m,
1
3
44 1H), 7.19 (s, 1H), 7.13-7.09 (m, 3H), 6.36 (s, 1H), 5.12 (s, 2H), 2.17 (s, 3H). C NMR (150 MHz,
45 DMSO-d ) δ: 166.9, 166.4, 162.9, 159.8, 145.0, 144.0, 140.9, 135.4, 134.5, 132.3, 128.5, 126.8,
6
2
5

ACCEPTED MANUSCRIPT
+
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
17 4 3
46 126.3, 123.8, 122.8, 119.5, 109.3, 103.9, 35.2, 10.9. HRMS calcd for C20H N O (M+H)
47 361.1300, found 361.1295.
48
49 4.1.1.27. (E)-2-(5-Methoxy-1-(4-nitrobenzyl)-2-oxoindolin-3-ylidene)-N-(quinolin-6-yl)acetamide
o
1
50 (31). Yield: 79%. Mp = 230-232 C. H NMR (600 MHz, DMSO-d ) δ: 11.22 (s, 1H), 8.81 (dd, J
6
51 = 4.2, 1.8 Hz, 1H), 8.61 (d, J = 2.4 Hz, 1H), 8.40 (d, J = 3.0 Hz, 1H), 8.38 (dd, J = 8.4, 0.6 Hz,
52 1H), 8.20 (dt, J = 9.6, 2.4 Hz, 2H), 8.01 (d, J = 9.0 Hz, 1H), 7.88 (dd, J = 9.0, 2.4 Hz, 1H), 7.58 (d,
53 J = 9.6 Hz, 2H), 7.50 (dd, J = 8.4, 4.2 Hz, 1H), 7.31 (s, 1H), 6.96 (dd, J = 8.4, 2.4 Hz, 1H), 6.87 (d,
1
3
54 J = 9.0 Hz, 1H), 5.11 (s, 2H), 3.74 (s, 3H). C NMR (150 MHz, DMSO-d ) δ: 167.2, 162.8, 155.3,
6
55 149.4, 146.9, 145.1, 144.2, 137.9, 136.6, 135.8, 135.1, 129.8, 128.3, 127.6, 123.9, 123.3, 121.9,
+
56 120.6, 116.6, 115.8, 115.6, 109.7, 55.7, 42.4. HRMS calcd for C27
H
21
N
4
O
5
(M+H) 481.1512,
57 found 481.1527.
58
59 4.1.1.28.
60 (E)-2-(1-(4-Aminobenzyl)-5-methoxy-2-oxoindolin-3-ylidene)-N-(quinolin-6-yl)acetamide
(32).
1
61 Yield: 78%. Amorphous powder. H NMR (600 MHz, DMSO-d
6
) δ: 11.19 (s, 1H), 8.81 (dd, J =
62 4.2, 1.8 Hz, 1H), 8.61 (d, J = 2.4 Hz, 1H), 8.40 (d, J = 3.0 Hz, 1H), 8.39 (d, J = 8.4 Hz, 1H), 8.01
63 (d, J = 9.0 Hz, 1H), 7.88 (dd, J = 9.0, 2.4 Hz, 1H), 7.50 (dd, J = 8.4, 4.2 Hz, 1H), 7.26 (s, 1H),
64 7.00 (d, J = 8.4 Hz, 2H), 6.95 (dd, J = 8.4, 3.0 Hz, 1H), 6.87 (d, J = 8.4 Hz, 2H), 6.49 (d, J = 8.4
1
3
6
65 Hz, 2H), 5.03 (br, 2H), 4.73 (s, 2H), 3.73 (s, 3H). C NMR (150 MHz, DMSO-d ) δ: 166.9, 162.9,
66 155.0, 149.4, 148.1, 145.0, 138.5, 136.6, 135.8, 135.5, 129.8, 128.4, 128.3, 127.1, 123.3, 122.8,
+
67 121.9, 120.5, 116.5, 115.6, 115.5, 113.9, 110.0, 55.7, 42.7. HRMS calcd for C H N O (M+H)
2
7
23
4
3
2
6

ACCEPTED MANUSCRIPT
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
68 451.1770, found 451.1777.
69
70 4.1.1.29.
71 (E)-2-(1-(4-Bromobenzyl)-5-methoxy-2-oxoindolin-3-ylidene)-N-(quinolin-6-yl)acetamide
(33).
o
1
72 Yield: 78%. Mp = 201-203 C. H NMR (600 MHz, DMSO-d ) δ: 11.21 (s, 1H), 8.81 (dd, J = 4.2,
6
73 1.2 Hz, 1H), 8.61 (d, J = 2.4 Hz, 1H), 8.44 (d, J = 3.0 Hz, 1H), 8.39 (d, J = 7.8 Hz, 1H), 8.01 (d, J
74 = 9.0 Hz, 1H), 7.88 (dd, J = 9.0, 1.8 Hz, 1H), 7.53 (d, J = 8.4 Hz, 2H), 7.51 (dd, J = 4.2, 2.4 Hz,
75 1H), 7.28 (d, J = 8.4 Hz, 2H), 7.28 (s, 1H), 6.96 (dd, J = 8.4, 2.4 Hz, 1H), 6.87 (d, J = 8.4 Hz, 1H),
1
3
76 4.93 (s, 2H), 3.73 (s, 3H). C NMR (150 MHz, DMSO-d ) δ: 167.1, 162.8, 155.2, 149.4, 145.0,
6
77 138.0, 136.6, 135.8, 135.7, 135.2, 131.6, 129.8, 129.4, 128.3, 127.4, 123.3, 121.9, 120.6, 120.5,
+
78 116.6, 115.7, 115.6, 109.8, 55.7, 42.3. HRMS calcd for C27
H
21
N
3
O
3
Br (M+H) 514.0766, found
79 514.0778.
80
81 4.1.1.30.
82 (E)-2-(1-(4-Bromobenzyl)-5-hydroxy-2-oxoindolin-3-ylidene)-N-(quinolin-6-yl)acetamide
(34).
o
1
83 Yield: 78%. Mp = 221-223 C. H NMR (600 MHz, DMSO-d
6
) δ: 11.54 (s, 1H), 9.51 (s, 1H), 8.81
84 (dd, J = 4.2, 1.8 Hz, 1H), 8.48 (d, J = 2.4 Hz, 1H), 8.38 (d, J = 7.8 Hz, 1H), 8.01 (d, J = 9.0 Hz,
85 1H), 7.86 (dd, J = 9.0, 2.4 Hz, 1H), 7.52 (d, J = 8.4 Hz, 2H), 7.53-7.51 (m, 1H), 7.32 (s, 1H), 7.29
1
3
86 (d, J = 8.4 Hz, 2H), 7.22 (d, J = 2.4 Hz, 1H), 6.77-6.73 (m, 1H), 4.87 (s, 2H). C NMR (150 MHz,
87 DMSO-d ) δ: 165.2, 163.4, 153.5, 148.9, 144.4, 136.8, 136.1, 135.7, 134.6, 131.6, 131.5, 131.2,
6
88 129.7, 129.6, 129.5, 129.3, 129.2, 128.4, 123.7, 122.5, 121.9, 120.6, 117.4, 115.4, 110.2, 109.1,
+
89 42.1. HRMS calcd for C H N O Br (M+H) 500.0609, found 500.0603.
2
6
19
3
3
27

ACCEPTED MANUSCRIPT
5
90
91 4.1.1.31. (Ε)-2-(1-Methyl-2-oxoindolin-3-ylidene)-N-phenylacetamide (35). Yield: 78%. Mp =
5
o
1
5
5
5
5
5
5
5
5
6
6
6
6
6
6
6
6
6
6
6
6
92 236-238 C. H NMR (600 MHz, DMSO-d
6
) δ: 10.83 (s, 1H), 8.65 (d, J = 7.8 Hz, 1H), 7.75 (d, J
93 = 7.8 Hz, 2H), 7.41 (dt, J = 7.8, 0.6 Hz, 1H), 7.36 (t, J = 7.8 Hz, 2H), 7.15 (s, 1H), 7.11 (t, J = 7.8
1
3
94 Hz, 1H), 7.07 (t, J = 7.8 Hz, 1H), 7.03 (d, J = 7.8 Hz, 1H), 3.18 (s, 3H). C NMR (150 MHz,
95 DMSO-d ) δ: 167.2, 162.6, 145.6, 138.8, 134.8, 132.1, 128.9, 128.4, 126.9, 124.1, 122.3, 119.6,
6
96 119.4, 108.8, 26.1.
97
o
98 4.1.1.32. (E)-2-(2-Oxoindolin-3-ylidene)-N-phenylacetamide (36). Yield: 74%. Mp = 242-245 C.
1
99
H NMR (600 MHz, DMSO-d ) δ: 10.79 (s, 1H), 10.70 (s, 1H), 8.60 (d, J = 7.2 Hz, 1H), 7.74 (dd,
6
00 J = 8.4, 1.2 Hz, 2H), 7.36 (t, J = 7.8 Hz, 2H), 7.31 (dt, J = 7.8, 1.2 Hz, 1H), 7.10 (t, J = 7.8 Hz,
1
3
01 1H), 7.10 (s, 1H), 6.99 (dt, J = 7.8, 1.2 Hz, 1H), 6.86 (d, J = 7.8 Hz, 1H). C NMR (150 MHz,
02 DMSO-d ) δ: 168.5, 162.7, 144.5, 138.8, 135.7, 132.1, 128.9, 128.6, 126.3, 124.0, 121.7, 120.2,
6
+
03 119.4, 110.0. HRMS calcd for C H N O (M+H) 265.0977, found 265.0972.
1
6
13
2
2
04
05 4.1.1.33. (E)-2-(2-Oxoindolin-3-ylidene)-N-(quinolin-6-yl)acetamide (37). Yield: 68%. Mp =
o
1
06 165-168 C. H NMR (600 MHz, DMSO-d ) δ: 11.14 (s, 1H), 10.75 (s, 1H), 8.79 (dd, J = 4.2, 1.2
6
07 Hz, 1H), 8.65 (d, J = 7.2 Hz, 1H), 8.60 (d, J = 2.4 Hz, 1H), 8.34 (d, J = 8.4 Hz, 1H), 8.00 (d, J =
08 9.0 Hz, 1H), 7.85 (dd, J = 9.0, 2.4 Hz, 1H), 7.49 (dd, J = 8.4, 4.2 Hz, 1H), 7.33 (dt, J = 7.8, 1.2 Hz,
1
3
09 1H), 7.11 (s, 1H), 7.01 (dt, J = 7.8, 1.2 Hz, 1H), 6.88 (d, J = 7.8 Hz, 1H). C NMR (150 MHz,
10 DMSO-d ) δ: 168.7, 163.3, 149.6, 145.1, 144.7, 136.9, 136.3, 136.0, 132.5, 129.8, 128.9, 128.5,
6
+
11 126.1, 123.5, 122.2, 122.0, 120.4, 115.7, 110.3. HRMS calcd for C H N O (M) 315.1007,
1
9
13
3
2
2
8