Synthesis of 4-anilinoquinazoline-derivative dual kinase inhibitors targeting EGFR and VEGFR-2

Full text of DOI:10.1002/bkcs.11348

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DOI: 10.1002/bkcs.11348
BULLETIN OF THE
K. C. Bang et al.
KOREAN CHEMICAL SOCIETY
Synthesis of 4-Anilinoquinazoline-Derivative Dual Kinase Inhibitors
Targeting EGFR and VEGFR-2
Keuk Chan Bang,†,k Tae Hun Song,†,‡,k Young Jin Park,†,‡ Jong Soo Lee,†,‡ Seungah Jun,‡ Seung
†,
Hyun Jung,‡ Young-Jin Chun,†, and Ha Hyung Kim
*
*
†College of Pharmacy, Chung-Ang University, Seoul 06974, Korea.
*E-mail: yjchun@cau.ac.kr; hahyung@cau.ac.kr
‡Department of Drug Discovery, Hanmi Research Center, Gyeonggi-do 18469, Korea
Received September 27, 2017, Accepted November 17, 2017, Published online December 27, 2017
Keywords: 4-Anilinoquinazoline, Dual inhibitor, Epidermal growth factor receptor, Vascular endothelial
growth factor receptor-2
The epidermal growth factor receptor (EGFR) and vascular
endothelial growth factor receptor-2 (VEGFR-2) signaling
pathway have been clinically validated in solid tumors.1,2
EGFR is highly overexpressed in a variety of cancers and
plays a role in enhanced cell proliferation, in escape from
apoptosis to ensure cancer cell survival, and finally, in the
aggressive growth of invasive tumors.3
VEGFR-2 is the principal receptor in angiogenesis. Inhi-
bition of VEGFR-2 signaling pathway influences tumor
growth and metastasis by inhibiting tumor angiogenesis.4,5
Although single-pathway inhibitors have shown potential
in cancer therapy, these pathways have shown limited clini-
cal efficacy because cancer has complex pathology.6,7
However, it is highly recommended to treat cancer with
dual-pathway inhibitors because of the heterogeneous char-
acteristics of cancer. In addition, dual kinase inhibitors
could synergistically inhibit tumor growth.8
In our study, a series of novel dual-acting compounds
were designed and synthesized to inhibit EGFR and
VEGFR-2. In vitro kinase profiling and cell-based screen-
ing together with Structure–Activity Relationship (SAR)
studies finally led to the discovery of compound 1, which
was a hybrid structure containing both acryl amide of CI-
1033 and 4-bromo-2-fluoroaniline moiety of ZD-6474
(Figure 1).9,10
modified to vary the number of carbons (5–7). All of the
modified compounds showed good IC50 values for EGFR
in the range of 2 to 10 nM; in particular, 7 (n = 1) dis-
played better VEGFR-2 activity than 5 (n = 3) and
6 (n = 2). Thus, the short chain was superior to the long
chains. Next, we evaluated the influence of R1-substituted
analogues, which were derived with various alkyl (7–10) or
hetero alkyl (11−13) substituents on EGFR and VEGFR-2,
for which the carbon chain was fixed at n = 1. The hetero
alkyl analogues (11 and 13) exhibited better inhibitory
activities than the alkyl analogues (7–10). Among the ana-
logues, 1112 showed stronger inhibition targeting EGFR
and VEGFR-2 than the other derivatives (Table 1). In addi-
tion, 11 demonstrated potent inhibitory activities against
mutated EGFRs (Table 2) as well as A431, VEGF-induced
HUVEC, and H1975 cell lines. However, 11 did not inhibit
Hs27, the human normal cell line (Table 3). Thus, 11
showed strong inhibitory activities toward EGFR and
VEGFR-2 overexpressed cells as well as 1st generation
EGFR inhibitor-resistant cell, which was expressed as
T790 M mutation of EGFR in non-small cell lung cancer.13
On the other hand, Iressa and ZD-6474 did not show any
inhibitory activity against EGFR T790 M mutated cell and
The general synthetic procedure for the preparation of
compound 1 derivatives is summarized in Scheme 1.11
A
substituted quinazoline 2 was reacted with a substituted ani-
line, and subsequent nucleophilic aromatic substitution by
N-Boc amino alcohol afforded the alkoxy quinazoline 3.
After deprotection of N-Boc, the carboxylic acid group was
coupled with an amine to provide the amide 4. Subsequent
reduction of the nitro group on compound 4 and acrylation
using acryloyl chloride yielded the target compounds 5–13.
The enzyme inhibitory activities of EGFR and VEGFR-2
are summarized in Table 1. In order to investigate the SAR,
the chain length (n) at site C-7 of the quinazoline was
Figure 1. Schematic showing the design of compound 1 for the
merged EGFR–VEGFR pharmacophore.
kThese authors contributed equally to this work.
Bull. Korean Chem. Soc. 2018, Vol. 39, 123–125
© 2017 Korean Chemical Society, Seoul & Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
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KOREAN CHEMICAL SOCIETY
Table 2. Inhibitory activities against mutated EGFRs.
IC50 (nM)
EGFRT790M
Compound
EGFR
EGFRT790M/L858R
11
2
11
3
Iressa
ZD6474
530
800
> 1,000
> 1,000
> 1,000
> 1,000
Scheme 1. (a) 4-bromo-2-fluoroaniline, iPrOH, reflux, 4 h;
(b) N-Boc amino alcohol, KOTMS, DMSO, rt., 4 h; (c) TFA,
CH2Cl2, rt., 0.5 h; (d) R1CO2H, EDCI, pyridine, rt., 4 h; (e) Fe,
AcOH, reflux, 2 h; (f ) acryloyl chloride, DMF, rt., 1 h.
enzymes. In addition, 11 significantly suppressed angiogen-
esis dose-dependently in mice because of VEGFR-2 inhibi-
tion (Figure 2).
In summary, we developed a series of compounds having
4-anilinoquinazoline as the key structure. The compounds
were synthesized and evaluated for dual inhibitory activities
against EGFR and VEGFR-2. Compound 11 showed excel-
lent inhibitory activities against kinases and cells in EGFR
and VEGFR-2 as well as T790 M mutant EGFR pathway.
In addition, 11 demonstrated anti-angiogenic effect. Thus,
compound 11 could serve as a guide for the development
of an EGFR and VEGFR-2 dual inhibitor.
Figure 2. Compound 11 inhibit angiogenesis in the mice Matrigel
Plug assay.14
Table 3. Inhibitory activities in cell-based assay for A431, VEGF-
induced HUVEC, H1975, and Hs27.
Table 1. Inhibitory kinase activities of derivatives for EGFR and
IC50 (nM)
VEGFR-2
Compound
A431
HUVEC
H1975
Hs27
Br
11
Iressa
ZD6474
14
45
142
93
> 1,000
43
130
> 1,000
> 1,000
> 1,000
> 1,000
> 1,000
O
HN
HN
O
F
N
6
7
H
R1
N
N
n
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