Synthesis and in vitro antitumor activity of 1-(3-dimethylamino)propyl indolin-2-one derivatives

Article abstract of DOI:10.1007/s00044-012-0170-3

A series of 1-(3-dimethylaminopropyl)indolin-2-one derivatives were designed and synthesized based on the structural features of TMP-20, LK-B030, and BX-517. These newly synthesized derivatives were evaluated for in vitro activity against five human cancer cell lines and three HUVECs. Results revealed that all of the target compounds 1a-h generally show potent activity against these cancer cell lines and higher selectivity on VEGF- and bFGF-stimulated HUVECs than HUVEC. In particular, 1f (IC₅₀s: 1.10-1.47 μM) is 1.8-6.0-fold more potent than sunitinib against MDA-MB-231, A549, HL-60 and K-562, and 1.6-2.8-fold more potent than LK-B030 against MDA-MB-231 and A549.

Full text of DOI:10.1007/s00044-012-0170-3

MEDICINAL
CHEMISTRY
RESEARCH
Med Chem Res
DOI 10.1007/s00044-012-0170-3
ORIGINAL RESEARCH
Synthesis and in vitro antitumor activity
of 1-(3-dimethylamino)propyl indolin-2-one derivatives
•
Kai Lv Li-Li Wang Xin-Bo Zhou Ming-Liang Liu
•
•
•
•
Hong-Ying Liu Zhi-Bing Zheng Song Li
•
Received: 4 January 2012 / Accepted: 28 June 2012
Ó Springer Science+Business Media, LLC 2012
Abstract A series of 1-(3-dimethylaminopropyl)indolin-
2-one derivatives were designed and synthesized based on
the structural features of TMP-20, LK-B030, and BX-517.
These newly synthesized derivatives were evaluated for in
vitro activity against five human cancer cell lines and three
HUVECs. Results revealed that all of the target compounds
1a–h generally show potent activity against these cancer
cell lines and higher selectivity on VEGF- and bFGF-
stimulated HUVECs than HUVEC. In particular, 1f (IC50s:
1.10–1.47 lM) is 1.8–6.0-fold more potent than sunitinib
against MDA-MB-231, A549, HL-60 and K-562, and
1.6–2.8-fold more potent than LK-B030 against MDA-
MB-231 and A549.
during the last decade. Indolin-2-one as a new scaffold in
this therapeutic area has been the subject of intense
research recently. Structural modifications mainly at the 3-
and 5-positions of the indolin-2-one ring have made con-
siderable progress in the ability to increase antitumor
activity through inhibition on different receptors (kamma-
sud et al., 2009; Lai et al., 2010). It was reported that
Semaxanib (Fig. 1) shows potent activity against vascular
endothelial growth factor (VEGF) receptor-1 and -2 tyro-
sine kinases (Shaheen et al., 1999), and sunitinib (Fig. 1), a
new multi-targeted receptor tyrosine kinase inhibitor (Sun
et al., 2003) with activity against many human cancer cell
lines (Young et al., 2010; Ikezoe et al., 2006; Lyros et al.,
2010), was approved for the treatment of advanced renal
cancer and gastrointestinal stromal tumor (GIST) after
disease progression on or intolerance to Imatinib mesylate
(Morabito et al., 2006). Islam et al. (2007a, b) reported that
BX-517 (Fig. 1), bearing an additional methyl substituent
on the alkene and a carbamido group at the 5-position of
the indolin-2-one ring, respectively, has single-digit nano-
molar activity against phosphoinositide-dependent kinase-1
(PDK1) and excellent selectivity against protein kinase A
(PKA). In the above studies, however, any group was not
placed at the N-1 position of the indolin-2-one scaffold.
As part of an ongoing program to find indolin-2-ones as
potential antitumor drug candidates, we have focused our
attention on introducing proper functional groups to the N-1
position of the indolin-2-one ring. Structural modifications
of the N-1 position have led to the discovery of two lead
compounds in our lab. Z24 (Fig. 1), a5-chloro-1-[(piperidin-
1-yl)methyl]indolin-2-one, was found to have good potency
in growth suppression of various types of tumors in vivo by
inhibiting angiogenesis in new blood vessels (Wang et al.,
2004; Lu et al., 2005; Lu et al., 2003). TMP-20 (Fig. 1), a
5-chloro-1-[3-(dimethylamino)propyl]indolin-2-one was
Keywords Indolin-2-ones ꢀ Synthesis ꢀ
Antitumor activity ꢀ VEGF
Introduction
Cancers still remain the second leading cause of death
worldwide after cardiovascular diseases (Lv et al., 2010).
Many classes of antitumor drugs with diverse structures
have been introduced in the market or under development
K. Lv ꢀ L.-L. Wang ꢀ X.-B. Zhou ꢀ H.-Y. Liu ꢀ
Z.-B. Zheng (&) ꢀ S. Li
Beijing Institute of Pharmacology and Toxicology,
Beijing 100850, People’s Republic of China
e-mail: zzbcaptain@yahoo.com.cn
K. Lv ꢀ M.-L. Liu (&)
Institute of Medicinal Biotechnology, Chinese Academy
of Medical Sciences and Peking Union Medical College, Beijing
100050, People’s Republic of China
e-mail: lmllyx@yahoo.com.cn
123

Med Chem Res
Y
R3
R2
4/
N
H
N
H
N
R1
H2N
N
X
N
H
H
O
Cl
H
O
O
N
O
O
N
H
N
N
H
N
N
Semaxanib (X = Y = H )
Sunitinib ( X = F, Y =
BX-517
Z-24
TMP-20 (R1 = R2 = R3 = H)
Et2N(CH2)2NHCO )
LK-B030 (R1 = R3 =Me, R2 =
Et2N(CH2)2NHCO)
Fig. 1 Structures of some indolin-2-ones
then confirmed to possess more chemical stability while
retaining the good antitumor activity of Z24. Most recently,
we designed and synthesized a series of 1-[(3-dimethyl-
amino)propyl]indolin-2-one derivatives by introduction of
linear and cyclic carbamoyl groups at the C-40 position of the
pyrrole ring of TMP-20, and found that LK-B030 (Fig. 1),
the most active compound of them, have more in vitro
antitumor activity than sunitinib against all of the tested
cancer cell lines (Lv et al., 2011).
presence of Zinc powder in diethyl ether at room temper-
ature (Scheme 1).
The target compounds 1a–h were easily obtained by
aldol condensation of 1-[3-(dimethylamino)propyl]-5-hal-
ogenated (F, Cl)-indolin-2-ones 10a, b (Lv et al., 2011;
David 1993) with pyrrole derivatives 5, 7 or 9a–c, and then
further hydrochlorate formation with hydrogen chloride gas
in diethyl ether (Scheme 2). The new synthesized com-
pounds were well characterized by 1H NMR, 13C NMR and
MS. Like LK-B030, the pyrrole-2-methylidene geometries
at the 3-position of indolin-2-one ring of the target com-
pounds (except for 1c and 1d), were confirmed to have the
Z configurations by NOE NMR experiments, which is the
same as expected (Lv et al., 2011).
Inspired by the above research results, a series of 1-[(3-
dimethylamino)propyl]-5-halogenated (F or Cl)-indolin-2-
one derivatives were designed and synthesized in this study
by introducing an alkyl (methyl or ethyl) or a phenyl group
on the alkene of TMP-20, or taking the place of the
substituted amine moiety at the C-40 position of the pyrrole
ring of LK-B030 with an ethoxyl or a morpholino group.
Our primary objective was to optimize the potency of these
compounds against human cancer cell lines and contribute
to the development of new antitumor agents, as well as add
new structure–activity relationship (SAR) information to
1-[3-(dimethylamino)propyl]-indolin-2-ones.
In vitro antitumor activity
All the newly synthesized 1-[(3-dimethylamino)propyl]in-
dolin-2-ones 1a–h were evaluated for in vitro antitumor
activity in five human cancer cell lines, including MDA-
MB-231 (breast cancer), A549 (lung adenocarcinoma),
HL-60 (promyelocytic leukemia), K-562 (erythromyelo-
blastoid), and IM-9 (B-lymphoblastoid) using standard
MTT assay (Green et al., 1984). The 50 % inhibition
concentrations (IC50s) were compared with those of LK-
B030 and sunitinib (Table 1).
Chemistry
The new pyrrole derivatives (5, 7, 9a–c) and novel indolin-2-
ones (1a–h) described herein were synthesized as shown in
Schemes 1 and 2, respectively. Coupling reaction of tert-
butyl acetoacetate 2 with sodium nitrite in acetic acid and
then reductive cyclization with ethyl acetoacetate gave bis-
ester 3. Selective hydrolytic decarboxylation and formyla-
tion of 3 yielded 5-formyl-pyrrole-3-carboxylate 5. Basic
hydrolysis of the ester 5 afforded the corresponding car-
boxylic acid 6, the latter was then amidated with morpho-
line in the presence of 1-ethyl-3-(3-dimethylaminopropyl)
carbodiimide hydrochloride (EDC) and N-hydroxybenzo-
triazole (HOBt) to obtain 3,5-dimethyl-4-(morpholine-4-
carbonyl)-1H-pyrrole-2-carbaldehyde 7 (Scheme 1).
The indolin-2-one derivatives 1a–h show potent in vitro
antitumor activity against the tested human cancer cell
lines. In the 5-fluoro-indolin-2-one series bearing a sub-
stituent on the alkene, methyl substitution is about 2-fold
more potent than ethyl analog (1a vs 1b), which is in
keeping with the activity profiles of BX-517, while phenyl
shows improvement over ethyl rather than methyl (1c vs
1b, 1a). In addition, replacement of the pyrrole moiety of
1c with a phenyl group displays less activity against the
cell lines except for MDA-MB-231 (1d vs 1c).
In the 5-halogenated-indolin-2-one series containing a
2,4-dimethylpyrrole moiety, their antitumor activity
depends mainly on the side chains at the C-40 position of
the pyrrole ring. For example, the esters (1e, f) are much
Pyrrole derivatives 9a–c were prepared conveniently by
Friedel-Crafts acylation reaction of pyrrole 8 with acetyl
chloride, propionyl chloride or benzoyl chloride in the
123

Med Chem Res
Scheme 1 Synthesis of 5, 7 and
9a–c
O
O
OC2H5
O
(a)
OC2H5
O
(c)
(b)
O
O
O
N
HO
N
N
O
O
O
H
H
2
3
4
O
O
O
OC2H5
N
OH
O
(e)
(f)
(d)
H
H
H
N
H
5
N
N
H
6
O
O
H
O
7
9a: R = CH3
9b: R = C2H5
9c: R = C6H5
R
(g)
N
H
9a-c
N
O
H
8
Reagents and conditions: (a) NaNO2, HOAc, rt, 4 h; (b) CH3COCH2CO2C2H5, Zn, HOAc, 60 oC, 3h, 51% (for two
steps); (c) HCl, C2H5OH, H2O, ref., 4h, 85%; (d) POCl3, DMF, Cl(CH2)2Cl, ref., 3 h, 85%; (e) KOH, CH3OH, H2O,
ref., 4 h, 93%; (f) HOBt, EDC, morpholine, DMF, rt., 12 h, 46%; (g) RCOCl, Zn, (C2H5)2O, 40-65%
Scheme 2 Synthesis of indolin-
2-one derivatives 1a–h
R
Z
X
X
1) 5, 7, 9a-c, pyrrolidine, C2H5OH, ref., 2h
O
.
HCl
O
N
2) HCl, (C2H5)2O, rt, 0.5h
41-59%
N
N
N
10a: X = F
10b: X = Cl
1a-h
C2H5
O
O
H3C
CH3
N
H
1a: X = F, R = CH3, Z =
1b: X = F, R = C2H5, Z=
1c: X = F, R = C6H5, Z=
1d: X = F, R = C6H5, Z= C6H5
1e: X = F, R = H, Z =
1f: X = Cl, R = H, Z =
1g: X = F, R = H. Z =
1h: X =Cl, R = H, Z=
N
H
C2H5
O
O
H3C
CH3
N
N
H
H
O
O
N
H3C
CH3
N
H
N
H
O
O
N
H3C
CH3
N
H
more active than the corresponding amides (1g, h).
Moreover, 5-chloroindolin-2-ones are generally more
active than 5-fluoro analogs (1f vs 1e, 1h vs 1g). It was
worth noting that both of the esters 1e and 1f (IC50s:
1.10–3.86 lM) are more potent than the reference drug
sunitinib (IC50s: 2.41–6.61 lM) against the cell lines
except for IM-9. In particular, the most active compound
1f (IC50s: 1.10–1.47 lM) was found to be 1.8–6.0-fold
more potent than sunitinib against MDA-MB-231, A549,
HL-60 and K-562, and 1.6–2.8-fold more potent than
LK-B030 (IC50s: 2.29–3.11 lM) against MDA-MB-231
and A549.
including a VEGF- and a recombinant human basic fibro-
blast growth factor (bFGF)-stimulated HUVEC using
standard MTT assay (Green et al., 1984) and the results are
reported in Table 1. Except for 1e which possesses less
potent inhibitory activity against VEGF-stimulated HU-
VEC (IC50: 10.09 lM) than HUVEC (IC50: 8.56 lM), all
other target compounds show greater inhibition against
VEGF-stimulated HUVEC (IC50s: 4.21–9.71 lM) and
bFGF-stimulated HUVEC (IC50s: 8.83–22.04 lM) than
HUVEC (IC50s: 13.86–42.76 lM). The results indicated
that the newly synthesized indolin-2-ones have gener-
ally higher selectivity on VEGF- and bFGF-stimulated
HUVECs than HUVEC. Further studies on related mecha-
nism of action and structural modifications are currently in
progress.
All of the indolin-2-one derivatives 1a–h were also fur-
ther examined for inhibitory activity against proliferation of
three human umbilical venous endothelial cells (HUVEC)
123

Med Chem Res
Table 1 In vitro activity of the target compounds 1a–h against selected cell lines
Cell lines
ICa50 (lM)
Compounds
X
R
Z
MDA-MB-231
A549
HL-60
K-562
IM-9
HUVEC
Normal
bFGF
8.83
VEGF
9.71
1a
1b
F
F
CH3
5.92
8.27
7.99
6.41
8.82
6.68
13.86
22.20
C2H5
10.62
16.85
10.31
10.97
9.75
9.57
1c
F
F
C6H5
C6H5
6.92
6.61
11.38
13.41
7.16
6.43
6.85
5.64
9.38
42.76
32.78
8.83
9.52
8.42
1d
C6H5
10.13
10.87
1e
1f
F
H
H
H
H
3.86
1.47
1.55
1.10
1.36
1.17
1.54
1.36
3.21
2.95
8.56
21.98
29.45
14.23
1.81
10.16
22.04
10.68
10.09
9.66
9.11
4.21
Cl
F
1g
1h
14.71
9.69
21.65
8.60
13.24
4.64
57.03
5.46
61.48
7.65
Cl
LK-B030
Cl
H
2.29
4.51
3.11
6.61
1.07
3.53
0.52
2.41
0.47
1.35
5.57
3.90
3.08
4.04
2.72
2.75
Sunitinib
a
IC50 values were determined by at least three separate tests and reported as mean values
Conclusion
potent than sunitinib against MDA-MB-231, A549, HL-60
and K-562, and 1.6–2.8-fold more potent than LK-B030
against MDA-MB-231 and A549. In addition, compounds
1a–h have higher selectivity on VEGF- and bFGF-stimu-
lated HUVECs than HUVEC.
In summary, based on the structural features of MTB-20
and LK-B030 (previously discovered compounds with
good antitumor activity in our lab) and BX-517 (a lead
compound with single-digit nanomolar activity against
PDK1 and excellent selectivity against PKA), a series of
novel 1-(3-dimethylaminopropyl)indolin-2-one derivatives
were designed, synthesized and characterized by 1H NMR,
13C NMR, and MS. These newly synthesized derivatives
were evaluated for in vitro activity against five human
cancer cell lines and three HUVECs. Our results revealed
that all of the target compounds 1a–h show potent activity
against these cancer cell lines, and the most active com-
pound 1f (IC50s: 1.10–1.47 lM) is 1.8–6.0-fold more
Experimental
Melting points were determined in open capillaries and are
1
uncorrected. H NMR 13C NMR spectra were determined
on a Varian Mercury-400 spectrometer in DMSO-d6 or
CDCl3 using tetramethylsilane as an internal standard.
Mass spectra were obtained from Micromass ZabSpechigh-
resolution magnetic mass spectrometer. The reagents were
123

Med Chem Res
all of analytical grade or chemically pure. TLC was per-
formed on silica gel plates (Merck, ART5554 60F254).
concentrated under reduced pressure, and the residue was
recrystallized from dichloromethane (400 ml) to afford the
title compound 5 (2.16 g, 85 %) as a white solid mp:
1
2-Tert-butyl 4-ethyl 3,5-dimethyl-1H-pyrrole-2,4-
dicarboxylate (3)
165–167 °C. H-NMR (400 MHz, DMSO-d6) d ppm: 9.77
(brs, 1H), 9.61 (s, 1H), 6.35 (s, 1H), 4.32 (q, 2H, J = 7 Hz),
2.57 (s, 3H), 2.55 (s, 3H), 1.37 (t, 3H, J = 7 Hz).
To a stirring solution of tert-butyl acetoacetate 2 (6.96 g,
44 mmol) in acetic acid (15 mL) was added dropwise 1 N
NaNO2 aqueous solution (48 mL, 48 mmol) at 0 °C. The
reaction mixture was stirred for 4 h at room temperature to
give a pale yellow solution.
5-Formyl-2,4-dimethyl-1H-pyrrole-3-carboxylic
acid (6)
To a stirring solution of 5 (1.94 g, 10 mmol) in methanol
(10 mL) and distilled water (40 mL) was added 5 N KOH
aqueous solution (20 mL). The reaction mixture was heated
to refluxing and stirred for 4 h. Cooled to room temperature,
the mixture was adjusted to pH 3 with 5 N HCl, and then
filtered. The precipitate was washed with distilled water and
dried in vacuo to yield the title compound 6 (1.55 g, 93 %)
as an off-white solid mp 280–283 °C. 1H NMR (400 MHz,
DMSO-d6) d ppm: 12.14 (s, 1H), 12.12 (brs, 1H), 9.60 (s,
1H), 2.45 (s, 3H), 2.41 (s, 3H).
A mixture of ethyl acetoacetate (5.21 g, 40 mmol) and
zinc powder (7.84 g, 120 mmol) in acetic acid (20 mL)
was stirred for 1 h at 60 °C, and then the above pale yellow
solution was added dropwise to the mixture. The reaction
mixture was stirred for 3 h at the same temperature and
filtered. The solid obtained was washed with distilled
water, and recrystallized from petroleum ether to give the
title compound 3 (5.16 g, 51 %) as a white solid. mp
127–129 °C. 1H NMR (400 MHz, CDCl3) d ppm: 8.73
(brs, 1H), 4.28 (q, 2H, J = 7 Hz), 2.53 (s, 3H), 2.50 (s,
3H), 1.57 (s, 9H), 1.35 (t, 3H, J = 7 Hz).
3,5-Dimethyl-4-(morpholine-4-carbonyl)-
1H-pyrrole-2-carbaldehyde (7)
Ethyl 2,4-dimethyl-1H-pyrrole-3-carboxylate (4)
To a mixture of 6 (0.50 g, 3 mmol) and morpholine (0.35 g,
4 mmol) in DMF (20 mL) were added EDC (4.5 mmol) and
HOBt (4.5 mmol), and then stirred for 12 h at room tem-
perature. The reaction mixture was diluted with ethyl ace-
tate (50 mL), washed successively with distilled water and
saturated saline solution, dried over anhydrous MgSO4, and
filtered. The filtrate was concentrated under reduced pres-
sure to give the crude product 7 as oil which was used
directly in the next step without further purification.
To a solution of 3 (2.67 g, 10 mmol) in ethanol (300 mL)
and distilled water (50 mL) was added concentrated HCl
(10 mL) at room temperature. The reaction mixture was
heated to refluxing and stirred for 4 h. Cooled to room
temperature the mixture was diluted with ice-water
(300 mL), adjusted to pH 12 with 3 N NaOH aqueous
solution and extracted with ethyl acetate (3 9 50 mL). The
combined extracts were dried over anhydrous MgSO4, and
then filtered. The filtrate was concentrated under reduced
pressure to afford the title compound 4 (1.41 g, 85 %) as a
yellow solid. mp 66–68 °C. 1H NMR (400 MHz, CDCl3) d
ppm: 8.01 (brs, 1H), 6.35 (s, 1H), 4.26 (q, 2H, J = 7 Hz),
2.49 (s, 3H), 2.23 (s, 3H), 1.34 (t, 3H, J = 7 Hz).
General procedure for the synthesis of 2-acylpyrroles
9a–c
A mixture of 8 (40 mmol), acyl chloride (35 mmol) and
zinc powder (80 mmol) in anhydrous diethyl ether (60 mL)
was stirred at 0 °C for 3 h, and then filtered. The filtrate
was concentrated under reduced pressure, and the residue
was purified by column chromatography (silica gel) eluted
with petroleum ether and ethyl acetate (v:v = 20:1) to
afford 9a–c as pale yellow solids (40–65 %). 9a, mp:
86–88 °C (mp: 88–89 °C) (Gayido et al., 1984). 9b, mp:
51–53 °C (mp: 51–52 °C) (Gayido et al., 1984). 9c, mp:
77–79 °C (mp: 76–77 °C) (Wallace et al., 1993).
Ethyl 5-formyl-2,4-dimethyl-1H-pyrrole-3-carboxylate (5)
To a stirring solution of phosphorus oxychloride (2.30 g,
15 mmol) in DMF (4 mL) was added dropwise a solution of 4
(2.16 g, 13 mmol) in 1,2-dichloroethane (100 mL) at room
temperature. The reaction mixture was heated to refluxing and
stirredfor3 h.Cooledtoroomtemperature, tothemixturewas
added dropwise 1 N NaOAc solution (75 ml) and then stirred
at reflux for another 0.5 h. The aqueous layer was separated,
adjusted pH 12 with 5 N NaOH aqueous solution and
extracted with dichloromethane (3 9 40 mL). The extracts
were combined with the above organic layer, and washed
successively with distilled water and saturated saline solution,
dried over anhydrous MgSO4, and filtered. The filtrate was
General procedure for the synthesis of indolin-2-one
derivatives 1a–h
To a mixture of 10a, b (Shaheen et al. 1999) (3.1 mmol)
and 5, 7 or 9a–c (3 mmol) in ethanol (15 mL) was added
123

Med Chem Res
pyrrolidine (0.3 mL). The reaction mixture was heated to
refluxing and stirred for 2 h, and then concentrated under
reduced pressure. The residue was purified by column
chromatography (silica gel) eluted with methanol and
dichloromethane (v:v = 1:40) to yield the free bases of
1a–h. To a solution of the above free bases dissolved in
absolute diethyl ether was pumped dried hydrochloride gas
at 0–5 °C for 0.5 h. The reaction mixture was allowed to
stir for another 0.5 h at room temperature. The resulting
solid was collected by suction, and dried in vacuo to give
the title compounds 1a–h (41–59 %, for two steps) as
yellow solids.
(brs, 1H), 14.42 (s, 1H). 13C NMR (400 MHz, DMSO-d6) d
ppm: 22.54, 37.20, 41.93, 54.12, 109.08 (d, J = 10 Hz),
109.23 (d, J = 27 Hz), 111.58, 112.43 (d, J = 23 Hz),
113.54, 122.27, 125.27 (d, J = 10 Hz), 126.76, 128.03,
128.73, 129.18, 131.90, 135.78, 138.99, 145.48, 157.51 (d,
J = 233 Hz), 167.45. EI-MS: m/z 389 M?. HRMS-ESI (m/
z): Calcd. for C24H25ON3F (M ? H)?: 390.1976, found:
390.1975.
1-(3-(Dimethylamino)propyl)-3-(diphenylmethylene)-5-
fluoroindolin-2-one hydrochloride (1d)
1
Mp: 248–250 °C. H NMR (400 MHz, DMSO-d6) d ppm:
(Z)-3-(1-(1H-pyrrol-2-yl)ethylidene)-1-(3-
(dimethylamino)propyl)-5-fluoroindolin-2-one
hydrochloride (1a)
1.92–1.96 (m, 2H), 2.71 (s, 3H), 2.72 (s, 3H), 3.06–3.09
(m, 2H), 3.72–3.75 (m, 2H), 5.73–5.76 (m, 1H), 7.09–7.11
(m, 2H), 7.31–7.37 (m, 7H), 7.55–7.57 (m, 3H), 9.78 (brs,
1H). 13C NMR (400 MHz, DMSO-d6) d ppm: 22.35, 36.65,
41.95, 54.12, 109.28 (d, J = 8 Hz), 109.56 (d, J = 27 Hz),
115.15 (d, J = 23 Hz), 123.30, 123.88 (d, J = 11 Hz),
127.64, 128.31, 129.04, 129.38, 129.48, 129.84, 138.38,
139.09, 140.61, 155.31, 157.25 (d, J = 235 Hz), 165.46.
EI-MS: m/z 400 M?. HRMS-ESI (m/z): Calcd. for
C26H26ON2F (M ? H)?: 401.2023, found: 401.2022.
1
Mp: 165–167 °C. H NMR (400 MHz, DMSO-d6) d ppm:
2.02–2.04 (m, 2H), 2.72 (s, 6H), 2.79 (s, 3H), 3.10–3.12
(m, 2H), 3.94–3.97 (m, 2H), 6.40–6.42 (m, 1H), 7.10–7.28
(m, 3H), 7.41–7.43 (m, 1H), 7.59–7.62 (m, 1H), 10.01 (brs,
1H), 14.75 (s, 1H). 13C NMR (400 MHz, DMSO-d6) d
ppm: 21.76, 22.60, 39.07, 41.97, 54.18, 108.87 (d,
J = 8 Hz), 111.49 (d, J = 27 Hz), 111.52, 112.02 (d,
J = 25 Hz), 113.84, 119.64, 125.89 (d, J = 10 Hz),
126.10, 131.83, 135.33, 145.31, 158.13 (d, J = 233 Hz),
167.17. EI-MS: m/z 327 M?. HRMS-ESI (m/z): Calcd. for
C19H23ON3F (M ? H)?: 328.1819, found: 328.1818.
(Z)-Ethyl-5-((1-(3-(dimethylamino)propyl)-5-fluoro-2-
oxoindolin-3-ylidene)methyl)-2,4-dimethyl-1H-pyrrole-3-
carboxylate hydrochloride (1e)
Mp: 179–180 °C.1H NMR (400 MHz, DMSO-d6) d ppm:
1.28–1.32 (t, 3H, J = 7 Hz), 2.03–2.06 (m, 2H), 2.50–2.59
(m, 6H), 2.71 (s, 3H), 2.72 (s, 3H), 3.10–3.13 (m, 2H),
3.89–3.93 (t, 2H, J = 7 Hz), 4.18–4.24 (q, 2H, J = 7 Hz),
7.04–7.20 (m, 2H), 7.81 (s, 1H), 7.87–7.90 (m, 1H), 10.41
(brs, 1H), 13.78 (s, 1H). 13C NMR (400 MHz, DMSO-d6) d
ppm: 11.39, 14.17, 14.55, 22.56, 37.11, 41.91, 53.99, 59.14,
106.21 (d, J = 27 Hz), 109.34 (d, J = 12 Hz), 112.57 (d,
J = 25 Hz), 113.59, 114.39, 119.04, 124.69, 126.03,
133.56, 134.89, 141.00, 158.70 (d, J = 234 Hz), 164.13,
167.44. EI-MS: m/z 413 M?. HRMS-ESI (m/z): Calcd. for
C23H29O3N3F (M ? H)?: 414.2187, found: 414.2185.
(Z)-3-(1-(1H-pyrrol-2-yl)propylidene)-1-(3-
(dimethylamino)propyl)-5-fluoroindolin-2-one
hydrochloride (1b)
1
Mp: 186–188 °C. H NMR (400 MHz, DMSO-d6) d ppm:
1.31–1.35 (t, 3H, J = 7 Hz), 2.00–2.02 (m, 2H), 2.73 (s,
6H), 3.09–3.17 (m, 4H), 3.93–3.97 (t, 2H, J = 7 Hz),
6.43–6.45 (m, 1H), 7.16–7.24 (m, 3H), 7.39–7.42 (m, 2H),
9.92 (brs, 1H), 14.71 (s, 1H). 13C NMR (400 MHz, DMSO-
d6) d ppm: 14.61, 22.54, 26.07, 37.15, 41.91, 54.14, 109.07
(d, J = 10 Hz), 110.49 (d, J = 27 Hz), 111.67, 112.26 (d,
J = 23 Hz), 112.91, 119.07, 124.49 (d, J = 10 Hz),
125.90, 131.00, 135.54, 150.87, 158.35 (d, J = 232 Hz),
167.57. EI-MS: m/z 341 M?. HRMS-ESI (m/z): Calcd. for
C20H25ON3F (M ? H)?: 342.1976, found: 342.1974.
(Z)-Ethyl-5-((5-chloro-1-(3-(dimethylamino)propyl)-2-
oxoindolin-3-ylidene)methyl)-2,4-dimethyl-1H-pyrrole-3-
carboxylate hydrochloride (1f)
1
(Z)-1-(3-(dimethylamino)propyl)-5-fluoro-3-(phenyl(1H-
pyrrol-2-yl)methylene) indolin-2-one hydrochloride (1c)
Mp: 261–263 °C. H NMR (400 MHz, DMSO-d6) d ppm:
1.29–1.32 (t, 3H, J = 7 Hz), 2.00–2.07 (m, 2H), 2.53 (s,
3H), 2.56 (s, 3H), 2.73 (s, 6H), 3.11 (brs, 2H), 3.90–3.94 (t,
2H, J = 7 Hz), 4.20–4.25 (q, 2H, J = 7 Hz), 7.19–7.28
(m, 2H), 7.88 (s, 1H), 8.12 (d, 1H, J = 2 Hz), 10.11 (s,
1H), 13.75 (s, 1H). 13C NMR (400 MHz, DMSO-d6) d
ppm: 11.43, 14.25, 14.64, 22.61, 37.13, 41.98, 54.01,
59.24, 110.00, 113.80, 118.89, 125.16, 125.92, 126.19,
1
Mp: 232–234 °C. H NMR (400 MHz, DMSO-d6) d ppm:
2.04–2.09 (m, 2H), 2.73 (s, 6H), 3.14–3.16 (m, 2H),
3.96–3.99 (m, 2H), 4.86–4.89 (m, 1H), 5.91–5.93 (m, 1H),
6.30–6.32 (m, 1H), 6.96–6.99 (m, 1H), 7.15–7.17 (m, 1H),
7.31–7.33 (m, 2H), 7.50 (s, 1H), 7.60–7.63 (m, 3H), 10.59
123

Med Chem Res
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(PDK1) inhibitors. Part 1: design, synthesis and biological
activity. Bioorg Med Chem Lett 17:3814–3818
126.36, 126.53, 133.99, 137.35, 141.26, 164.18, 167.38.
FAB-MS: m/z 430 (M ? H)?. HRMS-ESI (m/z): Calcd. for
C23H29O3N3Cl (M ? H)?: 430.1892, found: 430.1896.
Islam I, Brown G, Bryant J, Hrvatin P, Kochanny MJ, Phillips GB,
Yuan S, Adler M, Whitlow M, Lentz D, Polokoff MA, Wu J,
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Bioorg Med Chem Lett 17:3819–3925
(Z)-3-((3,5-Dimethyl-4-(morpholine-4-carbonyl)-1H-
pyrrol-2-yl)methylene)-1-(3-(dimethylamino)propyl)-5-
fluoroindolin-2-one hydrochloride (1g)
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ketone compound Z24 in vivo and its anti-angiogenesis activity.
Chin J Pharmacol Toxicol 17:401–407
1
Mp: 267–269 °C. H NMR (400 MHz, DMSO-d6) d ppm:
2.02–2.06 (m, 2H), 2.29 (s, 3H), 2.32 (s, 3H), 2.72 (s, 6H),
3.09–3.13 (m, 2H), 3.50–3.58 (brs, 8H), 3.91–3.94 (t, 2H,
J = 7 Hz), 7.03–7.08 (m, 1H), 7.17–7.20 (m, 1H), 7.78 (s,
1H), 7.86–7.88 (m, 1H), 10.41 (brs, 1H), 13.54 (s, 1H). 13
C
NMR (400 MHz, DMSO-d6) d ppm: 10.13, 12.55, 22.70,
37.06, 42.03, 54.12, 66.44, 106.04 (d, J = 27 Hz), 109.40
(d, J = 10 Hz), 112.33 (d, J = 23 Hz), 113.03, 120.06,
125.14, 125.99, 126.29 (d, J = 10 Hz), 130.00, 134.75,
135.03, 158.66 (d, J = 233 Hz), 165.22, 167.55. FAB-MS:
m/z 455 (M ? H)?. HRMS-ESI (m/z): Calcd. for
C25H32O3N4F (M ? H)?: 455.2453, found: 455.2452.
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(Z)-5-Chloro-3-((3,5-dimethyl-4-(morpholine-4-carbonyl)-
1H-pyrrol-2-yl)methylene)-1-(3-
(dimethylamino)propyl)indolin-2-one hydrochloride (1h)
1
Mp: 271–273 °C. H NMR (400 MHz, DMSO-d6) d ppm:
2.02–2.06 (m, 2H), 2.30 (s, 3H), 2.32 (s, 3H), 2.71 (s, 3H),
2.72 (s, 3H), 3.09–3.13 (m, 2H), 3.38–3.58 (brs, 8H),
3.91–3.93 (t, 2H, J = 7 Hz), 7.22–7.27 (m, 2H), 7.83 (s,
1H), 8.07–8.09 (d, 1H, J = 2 Hz), 10.48 (s, 1H), 13.50 (s,
1H). 13C NMR (400 MHz, DMSO-d6) d ppm: 10.07, 12.51,
22.62, 37.07, 41.91, 54.00, 66.42, 109.91, 112.24, 118.51,
120.12, 125.27, 125.46, 126.09, 126.33, 126.54, 130.22,
135.12, 137.09, 165.18, 167.33. FAB-MS: m/z 471
(M ? H)?. HRMS-ESI (m/z): Calcd. for C25H32O3N4Cl
(M ? H)?: 471.2157, found: 471.2158.
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