Design, synthesis, and in vitro antiproliferative activity of novel Dasatinib derivatives

Article abstract of DOI:10.1016/j.bmcl.2011.12.070

Two series of novel Dasatinib derivatives have been designed and synthesized, with their in vitro cytostatic effect screened on human chronic myeloid leukemia cell line K562 and human myeloid leukemia cell line U937. Some target compounds demonstrated sig

Full text of DOI:10.1016/j.bmcl.2011.12.070

Bioorganic & Medicinal Chemistry Letters 22 (2012) 806–810
Contents lists available at SciVerse ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
Design, synthesis, and in vitro antiproliferative activity of novel
Dasatinib derivatives
⇑
Jin Cai, Shaoning Zhang, Ming Zheng, Xiaoqing Wu, Junqing Chen, Min Ji
School of Chemistry and Chemical Engineering, Institute of Pharmaceutical Engineering, Southeast University, 87 Dingjiaqiao, Nanjing, Jiangsu 210096, PR China
a r t i c l e i n f o
a b s t r a c t
Article history:
Received 15 June 2011
Revised 1 November 2011
Accepted 13 December 2011
Available online 20 December 2011
Two series of novel Dasatinib derivatives have been designed and synthesized, with their in vitro cyto-
static effect screened on human chronic myeloid leukemia cell line K562 and human myeloid leukemia
cell line U937. Some target compounds demonstrated significant inhibitory activities against both cell
lines. Compared to the contrast drug Dasatinib, 1b, 1c, 1d, 1e and 1f were found to demonstrate more
potent antitumor activities. The structures of all the newly synthesized compounds were determined
by 1H NMR and 13C NMR.
Keywords:
Dasatinib
Ó 2011 Elsevier Ltd. All rights reserved.
Tyrosine kinase
Cytotoxic activity
Cancer cell
Leukemias are uniformly fatal diseases characterized by exces-
sive and abnormal proliferation of primitive white blood cells and
their precursors with infiltration into the various tissues of the
body.1 Chronic myelogenous leukemia (CML) is a hematological
disorder caused by a chromosomal rearrangement that generates
a fusion protein, Bcr-Abl, with deregulated tyrosine kinase activ-
ity,2 which is crucial for malignant transformation in CML. The rec-
ognition of the Bcr-Abl gene and corresponding protein led to the
synthesis of small-molecule drugs, designed to interfere with
Bcr-Abl tyrosine kinase activation by competitive binding at the
ATP-binding site.3
(Fig. 1), are three selective EGFR inhibitors approved by the FDA
in 2003, 2004 and 2007 for locally advanced or metastatic tumor
therapy.12–14 They possessed the common structure character: 4-
anilino quinazoline moiety. On the basis of previous work in our
laboratory,15according to structure–activity relationship analysis
of the leading compound Dasatinib, as well as the basic drug design
principals of ring-addition and combination, we have devised and
synthesized two series of 4-benzothienyl amino quinazolines and
4-benzothienyl pyrimidines derivatives (Fig. 2): series 1, replace-
ment of pyrimidine with quinazoline, and thiazole ring with ben-
zothiophene; series 2, replacement of 4-hydroxyethylpiperazine
moiety with the other polar substituents, and thiazole ring with
benzothiophene. Our objective was to determine whether these
compounds favor greater inhibition of cell proliferation and higher
induction of cell death. The antitumor effect of all the newly syn-
thesized compounds on the in vitro growth of two cell lines,
namely human chronic myeloid leukemia cell line K562 and hu-
man myeloid leukemia cell line U937, was evaluated. Apparent
growth inhibition was observed for most compounds, with 1b,
1c, 1d, 1e and 1f demonstrating more potent activities against
K562 and U937 as compared to Dasatinib, respectively. Further-
more, some structure–activity relationships have also been
established.
Imatinib mesylate (GleevecÒ, the first Bcr-Abl tyrosine kinase
inhibitor (TKI), is one of the most well-known molecularly targeted
therapeutics and has revolutionized treatment of CML.4,5 However,
some patients initially treated with imatinib will need alternative
therapy, due to drug resistance, which is often caused by the
appearance of clones expressing mutant forms of Bcr-Abl.3
These issues have fueled the development of a number of next-
generation Bcr-Abl inhibitors.6,7 Second-generation TKI Dasatinib
(BMS-354825, SPRYCELÒ is a high affinity dual Src/Abl and c-Kit
inhibitor approved in the US and European Union for the treatment
of CML as well as Philadelphia chromosome-positive acute lym-
phocytic leukemia (ALL).8,9 Dasatinib exhibits greater potency than
imatinib and inhibits the majority of kinase mutations in imatinib-
resistant CML.10,11
As summarized in Table 1, up to 12 compounds16 in series 1
(1a–1f) and series 2 (2a–2f) were synthesized, as illustrated in
Schemes 1–3.
The 4-anilino quinazolines derivatives, Gefitinib (ZD1839,
IressaÒ, Erlotinib (OSI-774, TarcevaÒ and Lapatinib (TykerbÒ
Synthesis of the key intermediate 7 of the two series com-
pounds is shown in Scheme 1. Cyclization of 2-chloro-5-nitrobenz-
aldehyde 3 with ethyl thioglycolate 2 gave the benzothiophene
compound 4, which was hydrolyzed to the corresponding
⇑
Corresponding author. Tel.: +86 025 83272078.
E-mail address: jimin@seu.edu.cn (M. Ji).
0960-894X/$ - see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmcl.2011.12.070

J. Cai et al. / Bioorg. Med. Chem. Lett. 22 (2012) 806–810
807
F
HN
O
HN
Cl
O
O
N
O
O
O
O
N
N
N
N
Erlotinib
Gefitinib
O
F
HN
N
Cl
H
N
N
O
S
O
O
Lapatinib
Figure 1. Chemical structure of Gefitinib, Erlotinib and Lapatinib.
O
S
N
R
N
Replacement
CH3
N
H
N
Replacement
HN
N
S
O
N
Cl
Dasatinib
N
CH3
N
HO
Cl
O
Cl
O
S
N
S
H
N
CH3
H
HN
CH3
HN
N
N
O
N
R
N
CH3
R
2a~2f
1a~1f
Figure 2. A design for synthesis of Dasatinib derivatives (1a–1f and 2a–2f).
carboxylic acid 5. Acyl chlorination of 5 with oxalyl chloride and
condensation with 2-chloro-6-methylbenzenamine afforded 6,
the nitro group of which was then reduced with hydrogen by using
ammonium formate and Pd/C as catalyst to produce the key inter-
mediates 7 in high yield (92%).
vitro. The cell viability was determined by the MTT-based assay
using cell proliferation reagent WST-8, a reagent solution prepared
as an aqueous solution containing 5 mM WST-8 (Sigma), 0.2 mM
1-methoxyphenazinium salt (Sigma) and 150 mM NaCl.17 Briefly,
the tumor cell lines in RPMI1640 medium with 10% fetal bovine
serum were plated in 96-well microtiter plates (5.0 Â 103 cells/
well), and allowed to adhere at 37 °C with 5% CO2 for 4 h. The test
compound was then added, and the cells were incubated at 37 °C
with 5% CO2 for 72 h later. Subsequently, cell growth medium
was removed, and WST-8 was added to each well for another incu-
bation of 1.5 h at 37 °C. Absorbance was finally measured with a
plate reader at 450 nm with correction at 650 nm. The results were
expressed as the percentage of absorbance of treated wells versus
that of vehicle control. IC50, the drug concentration causing 50%
growth inhibition, was calculated via sigmoid curve fitting using
GraphPad Prism 5.0.
Methyl 4-hydroxy-3-methoxybenzoate 8 as starting material
was alkylated with 1-bromo-3-chloropropane to give 9 in 94%
yield. Nitration of 9 with nitric acid in acetic acid afforded 10,
which was then reduced by powdered iron in acetic acid to give
11 in satisfactory yield (80%). In contrast, catalytic hydrogenation
using Raney/Ni or 5% Pd/C gave incomplete conversions, even after
a long reaction time. Cyclization of 11 with formamidine acetate
and chlorination with thionyl chloride gave 13. Aminolysis of 13
was performed using intermediate 7 to afford 14 in 94% yield.
The final step was nucleophilic displacement of the chlorine atom
with different aliphatic amines to yield the corresponding target
compounds series 1a–1f (Scheme 2). Starting from the 4, 6-di-
chloro-2-methylpyrimidine 15, series 2a–2f could be obtained
via the same synthetic methods as described above (Scheme 3).
Preliminary cytotoxic activity of these Dasatinib derivatives
(1a–1f, 2a–2f) on K562 and U937 cell lines was investigated in
The in vitro growth inhibition activities of the newly synthesized
Dasatinib derivatives 1a–1f and 2a–2f against cancer cell lines
K562 and U937 were first evaluated by the MTT-based assay using
Dasatinib as a positive control at the concentration of 10
lM. As
shown in Table 2, almost all the series 1 compounds demonstrated

808
J. Cai et al. / Bioorg. Med. Chem. Lett. 22 (2012) 806–810
Table 1
Structure of compounds 1a–1f and 2a–2f.
Cl
O
Cl
O
S
N
H
S
N
H
CH3
CH3
HN
HN
N
O
N
R
N
R
N
CH3
series 1
series 2
Compd
R
Mp (°C)
163–164
168–169
Compd
2a
R
Mp (°C)
160–162
222–224
1a
1b
O
O
O
N
O
N
N
N
O
N
N
N
2b
CH3
1c
144–145
2c
157–158
OH
1d
1e
O
O
N
N
159–161
155–157
2d
2e
N
N
N
N
CH2CH3
138–140
154–156
N
CH3
OH
H3C
N
CH3
1f
153–155
2f
211–212
O
N
evident anti-proliferation effects with inhibition percentages high-
er than Dasatinib for K562 and U937 except for 1a, it is worth point-
ing out that significant inhibition was achieved for compound 1f
with an inhibition percentage higher than 80%. Series 2 compounds
were relatively less potent than series 1, but 2c, 2d and 2e displayed
the equivalent inhibition percentage compared with Dasatinib.
After a close inspection, we found that series 1 compounds
showing potent cytotoxic activities belong to 4-amino quinazolins,
while series 2 compounds belong to pyrimidines. Moreover, the
cytotoxic effect of series 2 compounds was also consistently lower
than that of series 1. 4-anilino quinazoline moiety is the common
structure character of some EGFR inhibitors, so we speculated the
more potent antitumor activity of series 1 may contribute to the
multi-targets kinase inhibition of EGFR, Src/Abl and c-Kit. In series
1, 1f with methylpiperidine substitution demonstrated the most
strong cytotoxic activity, and the piperidine substituted 1b was
lower than 1f, so we thought that conformational restriction of
methyl group in piperidine ring would help to improve the antitu-
mor activity. Compounds with piperazine substituted like 2c, 2d
and 2e were more cytotoxic than the others, we speculated that
the introduction of piperazine substituents in series 2 could
improve cytotoxicities to various extents. The morpholine substi-
tuted compounds 1a and 2a both showed poor cytotoxic activity,
we thought that may be induced by the electro negativity of the
oxygen atoms.
The compounds that demonstrated evident growth inhibition
activities have been further assayed for their antitumor potency
indicated by IC50 values, which were calculated by linear regression
Table 2
In vitro cytotoxic activity of target compounds against K562 and U937 cells.
b,c
Compd
Growth inhibition percentage of cancer cell linea,b (%)
U937
IC50
(lM)
K562
K562
NT
4.01 1.51
8.97 2.49
3.55 0.61
9.88 2.34
2.15 0.63
NT
U937
1a
1b
1c
1d
1e
1f
2a
2b
2c
2d
2e
2f
9.99 1.24
71.79 5.31
59.79 4.12
72.52 2.11
51.16 6.10
84.21 8.23
13.86 2.25
8.78 0.90
56.17 4.00
75.71 4.35
76.93 3.56
77.98 6.22
75.98 3.17
80.25 3.99
8.37 1.02
20.64 3.10
40.60 2.91
46.46 5.03
37.12 3.11
24.55 2.84
37.22 2.61
9.09 3.11
4.34 1.15
4.25 2.19
4.01 1.27
3.48 0.59
3.45 0.63
NT
20.00 3.19
14.98 2.98
13.12 5.13
13.70 3.19
20.55 3.38
13.63 2.25
NT
32.39 2.58
33.92 5.12
27.80 1.99
20.47 2.76
37.39 1.79
17.03 4.53
16.87 2.10
21.90 4.01
22.15 3.31
12.83 2.07
Dasatinib
NT: no test.
a
Compounds tested at a concentration of 10
l
M.
Values are averages of three independent experiments.
IC50 is the concentration of compound required to inhibit the cell growth by 50% compared to an untreated control.
b
c

J. Cai et al. / Bioorg. Med. Chem. Lett. 22 (2012) 806–810
809
O2N
CHO
Cl
O2N
O2N
(ii)
(i)
S
CO2Et
S
COOH
3
4
5
NO2
NH2
CH3
CH3
H
N
H
N
(iv)
(iii)
S
S
O
O
Cl
Cl
7
6
Scheme 1. Synthesis of 5-amino-N-(2-chloro-6-methylphenyl)benzo[b]thiophene-2-carboxamide 7. Reagents and conditions: (i) HSCH2CO2Et, K2CO3, DMF, 4 h, 60 °C, 97%;
(ii) NaOH, THF, 6 h, rt, 96%; (iii) oxalyl chloride, 2-chloro-6-methylbenzenamine, triethylamine, DMF, THF, 6 h, 0 °C, 76%; (iv) HCOONH4, Pd/C, H2, CH3OH, 1.5 h, rt, 92%.
O
COOCH3
O
O
COOCH3
O
O
COOCH3
NO2
(iii)
(i)
(ii)
HO
Cl
Cl
8
9
10
Cl
N
O
N
O
COOCH3
NH2
O
O
O
(v)
(iv)
N
NH
Cl
O
Cl
O
Cl
13
12
11
Cl
Cl
O
O
S
S
N
H
N
(vi)
(vii)
H
CH3
CH3
HN
N
HN
N
7
O
O
N
Cl
O
N
R
14
1a-1f
Scheme 2. Synthetic route for the preparation of the target compounds 1a–1f. Reagents and conditions: (i) K2CO3, ClCH2CH2CH2Br, DMF, 4 h, 70 °C, 94%; (ii) HNO3, AcOH,
Ac2O, 6 h, 0–5 °C, 94%; (iii) Fe, AcOH, CH3COOC2H5, N2, 5 h, 50 °C, 80%; (iv) formamidine acetate, EtOH, 6 h, reflux, 95%; (v) SOCl2, DMF, 2 h, reflux, 83%; (vi) (CH3)2CHOH, 4 h,
reflux, 94%; (vii) KI, DMF, 0.5 h, 70 °C, 73–86%.
Cl
Cl
O
O
S
S
N
N
CH3
N
H
H
CH3
CH3
(ii)
HN
N
(i)
HN
N
N
7
N
N
Cl
Cl
R
CH3
2a-2f
Cl
CH3
16
15
Scheme 3. Synthetic route for the preparation of the target compounds 2a–2f. Reagents and conditions: (i) triethylamine, THF, 4 h, 50 °C, 86%; (ii) DMF, 2 h, 70 °C, 72–88%.
analysis of the concentration–response curves obtained for each
compound. It is evident from Table 2 that more than half of the
newly synthesized derivatives demonstrated apparent inhibitory
activities against both cell lines. Compared to the contrast drug
Dasatinib, 1b, 1c, 1d, 1e and 1f were found to be more potent anti-
tumor activities.
In conclusion, we have synthesized two series of novel Dasati-
nib derivatives and tested for their antitumor activities on K562
and U9371 cell lines. Preliminary anti-tumorigenic activities
indicated that half of the newly synthesized derivatives displayed
significant inhibitory activities against both cell lines. Moreover,
compounds 1b, 1c, 1d, 1e and 1f demonstrated more potent anti-
tumor activities when compared to the contrast drug Dasatinib.
From the structure–activity relationships we may conclude that
compounds containing 4-anilino quinazolin demonstrated signifi-
cantly better cytotoxic activities than those with pyrimidine. This
study may provide valuable information for further designing
and developing antitumor agents with potent activities.

810
J. Cai et al. / Bioorg. Med. Chem. Lett. 22 (2012) 806–810
7.88–7.90 (m, 2H, Ar–H), 8.06 (d, J = 8.8 Hz, 1H, Ar–H), 8.33 (s, 1H, –CH,
Acknowledgments
thiophene), 8.49 (s, 2H, Ar–H), 9.64 (s, 1H, –NH),10.29 (s, 1H, –NH–CO–); 13C
NMR (DMSO-d6, 75 MHz) d (ppm): 11.76, 18.20, 26.32, 46.43, 48.52, 56.27,
66.67, 102.00, 107.74, 108.75, 117.84, 122.57, 122.75, 125.94, 127.04, 128.38,
129.07, 132.28, 133.25, 135.44, 136.99, 138.64, 139.35, 139.61, 140.56, 146.95,
149.03, 152.79, 153.66, 156.40, 160.25; N-(2-chloro-6-methylphenyl)-5-(6-
methoxy-7-(3-(4-methylpiperazin-1-yl)propoxy)quinazolin-4-
This work is supported by National Basic Research Program of
China (No. 2011CB933503) and Technology Supporting Program
of Jiangsu province (BE2009639).
ylamino)benzo[b]thiophene-2-carboxamide (1e): 1H NMR (DMSO-d6,
500 MHz) d (ppm): 1.89–1.91 (m, 2H, –CH2–CH2O), 2.15 (s, 3H, Ar–CH3), 2.35
(s, 3H, –NCH3), 2.42–2.50 (m, 10H, 5 Â CH2N), 4.00 (s, 3H, -OCH3), 4.19 (t,
J = 6.4 Hz, 2H, –CH2O), 7.19 (s, 1H, Ar–H), 7.28–7.33 (m, 2H, Ar–H), 7.43 (dd,
J = 7.5 Hz,1H, Ar–H), 7.88–7.90 (m, 2H, Ar–H), 8.06 (d, J = 8.8 Hz, 1H, Ar–H),
8.33 (s, 1H, –CH, thiophene), 8.49 (s, 2H, Ar–H), 9.64 (s, 1H, –NH), 10.29 (s, 1H,
–NH–CO–); 13C NMR (DMSO-d6, 75 MHz) d (ppm): 11.65, 18.21, 26.00, 45.71,
52.70, 54.27, 54.74, 55.07, 56.25, 66.78, 98.92, 102.01, 107.80, 108.79, 117.85,
122.59, 122.75, 125.95, 127.05, 128.38, 129.08, 132.29, 135.45, 136.99, 138.65,
139.36, 146.95, 148.99, 152.81, 153.61, 156.40, 160.26; N-(2-chloro-6-
methylphenyl)-5-(6-methoxy-7-(3-(2-methylpiperidin-1-yl)propoxy)quinazolin-
4-ylamino)benzo[b]thiophene-2-carboxamide (1f): 1H NMR (DMSO-d6, 500 MHz)
d (ppm): 0.92–0.97 (d, 3H, –CHCH3), 1.18–1.28 (m, 2H, –CH2), 1.52–1.60 (m,
4H, –CH2), 1.90–1.92 (m, 2H, –CH2–CH2O), 2.12 (m, 2H, –CH2), 2.29 (s, 3H, Ar–
CH3), 2.40–2.54 (m, 1H, –CH), 2.81 (m,2H, –CH2–CH2–CH2–), 4.00 (s, 3H, -
OCH3), 4.20 (t, J = 6.4 Hz, 2H, –CH2O), 7.19 (s, 1H, Ar–H), 7.28–7.33 (m, 2H, Ar–
H), 7.43 (dd, J = 7.5 Hz, 1H, Ar–H), 7.88–7.90 (m, 2H, Ar–H), 8.06 (d, J = 8.8 Hz,
1H, Ar–H), 8.34 (s, 1H, –CH, thiophene), 8.49 (s, 2H, Ar–H), 9.64 (s, 1H, –NH),
10.29 (s, 1H, –NH–CO–); 13C NMR (DMSO-d6, 75 MHz) d (ppm): 18.21, 18.49,
23.40, 25.24, 25.75, 34.17, 45.68, 49.55, 51.48, 55.26, 56.28, 66.81, 102.02,
107.79, 108.77, 117.85, 122.58, 122.75, 125.94, 127.04, 128.38, 129.07, 132.29,
133.24, 135.44, 136.99, 138.64, 139.35, 139.61, 146.95, 149.03, 152.80, 156.40,
Supplementary data
Supplementary data associated with this article can be found, in
the online version, at doi:10.1016/j.bmcl.2011.12.070.
References and notes
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T.; Inigo, I.; Johnston, K.; Kamath, A.; Kan, D.; Klei, H.; Marathe, P.; Pang, S. H.;
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160.25;
N-(2-chloro-6-methylphenyl)-5-(2-methyl-6-morpholinopyrimidin-4-
ylamino)benzo[b]thiophene-2-carboxamide (2a): 1H NMR (DMSO-d6, 500 MHz)
d (ppm): 2.27 (s, 3H, –CH3), 2.35 (s, 3H, –CH3), 3.45 (t, 4H, –CH2), 3.67 (t, 4H, –
CH2), 5.85 (s, 1H, –CH, pyrimidine), 7.26–7.30 (m, 2H, Ar–H), 7.42 (dd,
J = 7.6 Hz, 1H,Ar–H), 7.58 (dd, J = 8.8 Hz,1H, Ar–H), 7.91 (d, J = 8.80 Hz, 1H, Ar–
H), 8.27 (s, 1H, Ar–H), 8.38 (d, 1H, Ar–H), 9.17 (s, 1H, –NH), 10.24 (s, 1H, –NH–
CO–) ; 13C NMR (DMSO-d6, 75 MHz) d (ppm): 14.02, 18.19, 43.98, 65.79, 81.54,
114.31, 120.41, 122.83, 126.01, 127.01, 128.30, 129.03, 132.25, 133.31, 133.61,
138.45, 138.61, 139.41, 139.68, 160.30, 161.33, 162.87, 165.60; N-(2-chloro-6-
methylphenyl)-5-(2-methyl-6-(piperidin-1-yl)pyrimidin-4-
9. Shah, N. P.; Tran, C.; Lee, F. Y.; Chen, P.; Norris, D.; Sawyers, C. L. Science 2004,
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ylamino)benzo[b]thiophene-2-carboxamide (2b): 1H NMR (DMSO-d6, 500 MHz) d
(ppm): 1.53 (t, 4H, –CH2), 1.61 (d, 2H, –CH2), 2.27 (s, 3H, –CH3), 2.33 (s, 3H, –
CH3), 3.51 (t, 4H, –CH2), 5.84 (s, 1H, –CH, pyrimidine), 7.26–7.32 (m, 2H, Ar–H),
7.41 (dd, J = 7.6 Hz, 1H,Ar–H), 7.58 (dd, J = 8.8 Hz, 1H, Ar–H), 7.91 (d, J = 8.8 Hz,
1H, Ar–H), 8.26 (s, 1H, Ar–H), 8.40 (d, 1H, - Ar–H), 9.10 (s, 1H, –NH), 10.23 (s,
1H, –NH–CO–); 13C NMR (DMSO-d6, 75 MHz) d (ppm): 18.19, 24.21, 24.91,
26.10, 44.50, 81.22, 113.99, 120.25, 122.78, 126.01, 127.01, 128.29, 129.03,
132.25, 133.33, 138.61, 138.70, 139.35, 139.69, 160.31, 161.32, 162.29, 165.58;
N-(2-chloro-6-methylphenyl)-5-(2-methyl-6-(4-methylpiperazin-1-yl)pyrimidin-
4-ylamino)benzo[b]thiophene-2-carboxamide (2c): 1H NMR (DMSO-d6,
500 MHz) d (ppm): 2.21 (s, 3H, –CH3), 2.26 (s, 3H, –CH3), 2.34 (s, 3H, –CH3),
2.37 (br, 4H, –CH2), 3.48 (br, 4H, –CH2), 5.84 (s, 1H, –CH, pyrimidine), 7.26–7.32
(m, 2H, Ar–H), 7.41 (dd, J = 7.6 Hz, 1H,Ar–H), 7.58 (dd, J = 8.8 Hz,1H, Ar–H), 7.91
(d, J = 8.8 Hz, 1H, Ar–H), 8.26 (s, 1H, Ar–H), 8.38 (s, 1H, – Ar–H), 9.12 (s, 1H, –
NH), 10.23 (s, 1H, –NH–CO–); 13C NMR (DMSO-d6, 75 MHz) d (ppm): 18.19,
26.04, 43.44, 45.69, 54.16, 81.53, 114.19, 120.36, 122.83, 126.01, 127.02,
128.31, 129.04, 132.25, 133.31, 133.51, 138.53, 139.38, 139.68, 160.30, 161.34,
162.61, 165.61; N-(2-chloro-6-methylphenyl)-5-(6-(4-ethylpiperazin-1-yl)-2-
methylpyrimidin-4-ylamino)benzo[b]thiophene-2-carboxamide (2d): 1H NMR
(DMSO-d6, 500 MHz) d (ppm): 1.04 t, 3H, –CH3), 1.18 (m, 2H, –CH2), 2.26 (s,
3H, –CH3), 2.35 (s, 3H, –CH3), 2.37 (br, 4H, –CH2), 3.48 (br, 4H, –CH2), 5.85 (s,
1H, –CH, pyrimidine), 7.26–7.32 (m, 2H, Ar–H), 7.41 (dd, J = 7.6 Hz, 1H,Ar–H),
7.58 (dd, J = 8.8 Hz,1H, Ar–H), 7.91 (d, J = 8.8 Hz, 1H, Ar–H), 8.26 (s, 1H, Ar–H),
8.39 (d, 1H, Ar–H), 9.12 (s, 1H, –NH), 10.23 (s, 1H, –NH–CO–); 13C NMR (DMSO-
d6, 75 MHz) d (ppm): 11.88, 18.19, 26.03, 43.58, 51.57, 51.96, 81.50, 114.16,
120.34,122.79, 126.01, 127.00, 128.29, 129.03, 132.25, 133.31, 133.48, 138.55,
138.60, 139.37, 139.67, 160.30, 161.31, 162.61, 165.59; N-(2-chloro-6-
methylphenyl)-5-(6-(4-(2-hydroxyethyl)piperazin-1-yl)-2-methylpyrimidin-4-
ylamino)benzo[b]thiophene-2-carboxamide (2e): 1H NMR (DMSO-d6, 500 MHz) d
(ppm): 2.26 (s, 3H, –CH3), 2.34 (s, 3H, –CH3), 2.43–2.50 (br, 6H, –CH2), 3.48 (br,
4H, –CH2), 3.54 (t, 2H, –CH2OH), 4.40 (s, 1H, -OH), 5.85 (s, 1H, –CH, pyrimidine),
7.26–7.32 (m, 2H, Ar–H), 7.41 (dd, J = 7.6 Hz, 1H,Ar–H), 7.58 (dd, J = 8.8 Hz,1H,
Ar–H), 7.91 (d, J = 8.8 Hz, 1H, Ar–H), 8.26 (s, 1H, Ar–H), 8.39 (d, 1H, Ar–H), 9.13
(s, 1H, –NH), 10.24 (s, 1H, –NH–CO–); 13C NMR (DMSO-d6, 75 MHz) d (ppm):
18.19, 26.04, 43.58, 52.76, 58.47, 60.19, 81.51, 114.14, 120.33, 122.80, 126.01,
127.00, 128.30, 129.03, 132.25, 133.30, 133.47, 138.55, 138.60, 139.37,139.67,
160.30, 161.31, 162.60, 165.59; N-(2-chloro-6-methylphenyl)-5-(2-methyl-6-(4-
methylpiperidin-1-yl)pyrimidin-4-ylamino)benzo[b]thiophene-2-carboxamide
(2f): 1H NMR (DMSO-d6, 500 MHz) d (ppm): 0.91 (d, 3H, –CH3), 1.20 (t, 1H, –
CH), 1.53–1.67 (br, 4H, –CH2), 2.26 (s, 3H,–CH3), 2.33 (s, 3H, –CH3), 3.48 (br, 4H,
–CH2), 5.85 (s, 1H, –CH, pyrimidine), 7.26–7.32 (m, 2H, Ar–H), 7.41 (dd,
J = 7.6 Hz, 1H,Ar–H), 7.58 (dd, J = 8.8 Hz,1H, Ar–H), 7.90 (d, J = 8.8 Hz, 1H, Ar–
H), 8.26 (s, 1H, Ar–H), 8.40 (d, 1H, Ar–H), 9.07 (s, 1H, –NH), 10.23 (s, 1H, –NH–
CO–) ; 13C NMR (DMSO-d6, 75 MHz) d (ppm): 18.18, 21.69, 26.08, 30.55, 33.14,
43.84, 81.32, 113.99,120.24, 122.77, 125.99, 127.00, 128.29, 129.03, 132.24,
133.33, 138.60, 139.33, 139.67, 160.30,161.33, 162.26, 165.60.
14. Kris, M. G.; Natale, R. B.; Herbst, R. S.; Lynch, T. J., Jr.; Prager, D.; Belani, C. P.;
Schiller, J. H.; Kelly, K.; Spiridonidis, H.; Sandler, A.; Albain, K. S.; Cella, D.; Wolf,
M. K.; Averbuch, S. D.; Ochs, J. J.; Kay, A. C. JAMA 2003, 290, 2149.
15. Wu, X.; Li, M.; Qu, Y.; Tang, W.; Zheng, Y.; Lian, J.; Ji, M.; Xu, L. Bioorg. Med.
Chem. 2010, 18, 3812.
16. Spectral data for 1a–1f and 2a–2f: N-(2-chloro-6-methylphenyl)-5-(6-
methoxy-7-(3-morpholinopropoxy)quinazolin-4-ylamino)benzo[b]thiophene-
2-carboxamide (1a): 1H NMR (DMSO-d6, 500 MHz) d (ppm): 1.97 (t, J = 6.9 Hz,
2H, –CH2), 2.28 (s, 3H, Ar–CH3), 2.40 (br, 4H, 2 Â CH2N), 3.17 (t, 2H, –CH2CH2N),
3.59 (t, J = 4.6 Hz, 4H, 2 Â CH2O), 4.00 (s, 3H, –OCH3), 4.20 (t, J = 6.4 Hz, 2H,
ÀCH2O), 7.21 (s, 1H, Ar–H), 7.28–7.33 (m, 2H, Ar–H), 7.43 (dd, J = 7.7 Hz, 1H,
Ar–H), 7.88–7.90 (m, 2H, Ar–H), 8.06 (d, J = 8.7 Hz, 1H, Ar–H), 8.33 (s, 1H, ÀCH,
thiophene), 8.50 (s, 2H, Ar–H), 9.64 (s, 1H, –NH), 10.29 (s, 1H, –NH–CO–); 13C
NMR (DMSO-d6, 75 MHz) d (ppm): 8.97, 18.20, 25.60, 53.32, 54.71, 56.25,
66.17, 66.72, 102.01, 107.82, 108.80, 117.85, 122.59, 122.74, 125.93, 127.04,
128.38, 129.07, 132.28, 133.25, 135.45, 136.98, 137.71, 138.64, 139.35, 139.62,
146.94, 148.99, 152.81, 153.58, 156.40, 160.25; N-(2-chloro-6-methylphenyl)-5-
(6-methoxy-7-(3-(piperidin-1-yl)propoxy)quinazolin-4-
ylamino)benzo[b]thiophene-2-carboxamide (1b): 1H NMR (DMSO-d6, 500 MHz) d
(ppm): 1.39 (m, 2H, –CH2–CH2–CH2), 1.49–1.53 (m, 4H, –CH2–CH2–CH2), 1.93–
1.97 (m, 2H,–CH2–CH2–CH2O), 2.29 (s, 3H, Ar–CH3), 2.40 (br, 4H, 2 Â CH2N),
2.42 (t, J = 7.1 Hz, 2H, –CH2CH2N), 4.00 (s, 3H, –OCH3), 4.20 (t, J = 6.5 Hz, 2H, –
CH2O), 7.19 (s, 1H, Ar–H), 7.28–7.33 (m, 2H, Ar–H), 7.43 (dd, J = 7.5 Hz, 1H, Ar–
H), 7.88–7.90 (m, 2H, Ar–H), 8.06 (d, J = 8.7 Hz, 1H, Ar–H), 8.33 (s, 1H, –CH,
thiophene), 8.48 (s, 2H, Ar–H), 9.64 (s, 1H, –NH), 10.29 (s, 1H, –NH–CO–); 13C
NMR (DMSO-d6, 75 MHz) d (ppm): 18.20, 24.09, 25.56, 26.04, 54.07, 54.99,
56.24, 66.86, 102.00, 107.79, 117.83, 122.57, 122.72, 125.93, 127.02, 128.37,
129.06, 135.43, 136.98, 138.64, 152.79, 156.38, 160.25; N-(2-chloro-6-
methylphenyl)-5-(7-(3-(ethyl(2-hydroxyethyl)amino)propoxy)-6-
methoxyquinazolin-4-ylamino)benzo[b]thiophene-2-carboxamide (1c): 1H NMR
(DMSO-d6, 500 MHz) d (ppm): 0.95–0.97 (m, 3H, –CH2–CH3), 1.88–1.93 (m, 2H,
–CH2–CH2–CH2), 2.29 (s, 3H, Ar–CH3), 2.46–2.54 (m, 4H, 2 Â CH2N), 2.61 (m, J =
6.9 Hz, 2H, –CH2CH2N), 3.45 (t, J = 6.4 Hz, 2H, –CH2OH), 4.00 (s, 3H, –OCH3),
4.20 (t, J = 6.4 Hz, 2H, –CH2O), 4.29 (s,1H, –OH), 7.20 (s, 1H, Ar–H), 7.28–7.33
(m, 2H, Ar–H), 7.43 (dd, J = 7.5 Hz, 1H, Ar–H), 7.88–7.90 (m, 2H, Ar–H), 8.06 (d, J
= 8.8 Hz, 1H, Ar–H), 8.34 (s, 1H, –CH, thiophene), 8.49 (s, 2H, Ar–H), 9.64 (s, 1H,
–NH), 10.29 (s, 1H, –NH–CO–); 13C NMR (DMSO-d6, 75 MHz) d (ppm): 11.82,
18.22, 26.51, 47.66, 49.77, 55.66, 56.27, 59.28, 66.66, 101.99, 107.77, 108.76,
117.85, 122.59, 122.75, 125.95, 127.05, 128.39, 129.08, 132.29, 133.26, 135.45,
137.00, 138.65, 139.36, 139.62, 146.95, 149.04, 152.79, 153.68, 156.41, 160.26;
N-(2-chloro-6-methylphenyl)-5-(7-(3-(diethylamino)propoxy)-6-
methoxyquinazolin-4-ylamino)benzo[b]thiophene-2-carboxamide (1d): 1H NMR
(DMSO-d6, 500 MHz) d (ppm): 0.92–0.97 (m, 6H, –CH2CH3), 1.89–1.91 (m, 2H, –
CH2–CH2O), 2.29 (s, 3H, Ar–CH3), 2.46–2.50 (m, 4H, 2 Â CH2N), 2.57 (t, J =
6.9 Hz,2H, –CH2–CH2–CH2–), 4.00 (s, 3H, –OCH3), 4.20 (t, J = 6.4 Hz, 2H, –CH2O),
7.19 (s, 1H, Ar–H), 7.28–7.33 (m, 2H, Ar–H), 7.43 (dd, J = 7.5 Hz, 1H, Ar–H),
17. Berridge, M. V.; Tan, A. S.; Herst, P. M. Biotechnol. Annu. Rev. 2005, 11, 127.