Combination of 4-anilinoquinazoline, arylurea and tertiary amine moiety to discover novel anticancer agents

Article abstract of DOI:10.1016/j.bmc.2015.12.001

In present study, 4-anilinoquinazolines scaffold, arylurea and tertiary amine moiety were combined to design, synthesize gefitinib analogs and discover novel anticancer agents. A series of 4-anilinoquinazoline derivatives (1, 2, 3 and 4) bearing arylurea and tertiary amine moiety at its 6-position were synthesized. Their antiproliferative activities in vitro were evaluated via MTT assay against A431 cell and A549 cell. The SAR of the title compounds was discussed. The compounds 2d, 2i and 2j with potent antiproliferative activities were evaluated their inhibitory activity against EGFR-TK. Compound 2j displayed potent inhibitory activity against EGFR-TK. In addition, compound 2j, at 50 mg/kg, can completely inhibit cancer growth in established nude mouse A549 xenograft model in vivo. These results suggest that the 4-anilinoquinazoline derivatives bearing diarylurea and tertiary amino moiety at its 6-position can serve as anticancer agents and EGFR inhibitors.

Full text of DOI:10.1016/j.bmc.2015.12.001

Accepted Manuscript
Combination of 4-anilinoquinazoline, arylurea and tertiary amine moiety to dis-
cover novel anticancer agents
Sai-Jie Zuo, Sai Zhang, Shuai Mao, Xiao-Xiao Xie, Xue Xiao, Min-Hnag Xin,
Wei Xuan, Yuan-Yuan He, Yong-Xiao Cao, San-Qi Zhang
PII:
S0968-0896(15)30169-3
DOI:
http://dx.doi.org/10.1016/j.bmc.2015.12.001
BMC 12694
Reference:
Bioorganic & Medicinal Chemistry
To appear in:
Received Date:
Revised Date:
Accepted Date:
6 October 2015
30 November 2015
1 December 2015
Please cite this article as: Zuo, S-J., Zhang, S., Mao, S., Xie, X-X., Xiao, X., Xin, M-H., Xuan, W., He, Y-Y., Cao,
Y-X., Zhang, S-Q., Combination of 4-anilinoquinazoline, arylurea and tertiary amine moiety to discover novel
anticancer agents, Bioorganic & Medicinal Chemistry (2015), doi: http://dx.doi.org/10.1016/j.bmc.2015.12.001
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Combination of 4-anilinoquinazoline, arylurea
and tertiary amine moiety to discover novel anticancer agents
Sai-Jie Zuo a, Sai Zhang b, Shuai Mao a, Xiao-Xiao Xie a, Xue Xiao b, Min-Hnag Xin a, Wei Xuan
a, Yuan-Yuan He a, Yong-Xiao Cao b, San-Qi Zhang a *
a
Department of Medicinal Chemistry, School of Pharmacy, Xian Jiaotong University, Xian,
Shaanxi, 710061, PR China
b Department of Pharmacology, School of Basic Medical Science, Xian Jiaotong University, Xian,
Shaanxi, 710061, PR China
*Corresponding author, E-mail address: sqzhang@mail.xjtu.edu.cn (S. Q. Zhang).
Abstract
In present study, 4-anilinoquinazolines scaffold, arylurea and tertiary amine
moiety were combined to design, synthesize gefitinib analogues and discover novel
anticancer agents. A series of 4-anilinoquinazoline derivatives (1, 2, 3 and 4) bearing
arylurea and tertiary amine moiety at its 6-position were synthesized. Their
antiproliferative activities in vitro were evaluated via MTT assay against A431 cell
and A549 cell. The SAR of the title compounds was discussed. The compounds 2d, 2i
and 2j with potent antiproliferative activities were evaluated their inhibitory activity
against EGFR-TK. Compound 2j displayed potent inhibitory activity against
EGFR-TK. In addition, compound 2j, at 50 mg/kg, can completely inhibit cancer
growth in established nude mouse A549 xenograft model in vivo. These results
suggest that the 4-anilinoquinazoline derivatives bearing diarylurea and tertiary amino
moiety at its 6-position can serve as anticancer agents and EGFR inhibitors.
Keywords gefitinib * diaryl urea * tertiary amine * EGFR-TK inhibitor * anticancer
agent
1

Introduction
The epidermal growth factor receptor (EGFR) belongs to the family of
protein-tyrosine kinase receptor (RTK), which plays an essential role in the regulation
of cell growth, differentiation, and survival.1 RTKs have been observed
over-expression and/or constitutive activation in numerous types of human tumor,
including colon, breast, ovarian, head and neck, and non-small cell lung cancers.
Among the known RTKs, EGFR and human epidermal growth factor receptor (HER2)
have been extensively studied and clinically validated as targets for chemical
therapies.2, 3 Thousands of small organic molecule inhibitors against EGFR or both
EGFR and HER2 were synthesized and evaluated. Gefitinib, erlotinib and lapatinib
were launched successfully to treat cancer in clinic (Figure 1).4, 5 The shared structure
of gefitinib, erlotinib and lapatinib is 4-anilinoquinazoline, whose structure activity
relationship has been elucidated.6
Figure 1. The structures of gefitinib, erlotinib and lapatinib
However, the efficacy of small organic molecules inhibitors such as gefitinib, etc.
is restricted to a small subset of patients due to molecular heterogeneity among and
within tumors.7, 8 The drug resistance caused by receptor mutation is another issue
needed to pay attention.9-11 Numbers of compounds with different structures were
discovered as EGFR or multi-target inhibitors.12-16 The essential pharmacophore for
2

4-anilinoquinazolines is the two nitrogen atoms at 1-posotion, 3-position and the
N-aryl at 4-position as well.17 The morpholine moiety in gefitinib is not involved in
any interaction with EGFR and is randomly ordered due to its lower electron density.
Consequently, the modification of the 6-substituted group in 4-anilinoquinazoline
with the group bearing active group provide the inhibitors with improved potency.18-20
Similarly, the replacement of 6-substituted group in gefitinib with acrylamido group
produced irreversible EGFR inhibitors21 and EGFR-T790M inhibitors.22
There are two hydrogen-bond donors and one hydrogen-bond acceptor in the urea
moiety. Aryl urea moiety is present in many classes of biologically active
compounds.23-25 Recent years, some kinase inhibitors with aryl urea moiety were
launched for the treatment of cancers or tested clinically. For example, linifanib,
developed by Abbott, is used for the treatment of colorectal cancer.26 Sorafenib,
developed by Bayer, is used for the treatment of advanced renal cell carcinoma.27
Meanwhile, urea moiety and 4-anilinoquinazolines scaffold were combined to search
for new compounds with antimalarial activity.28 The N-mustard pharmacophore was
attached to the 6-position of the 4-anilinoquinazolines via a urea linker to discover
dual functional antitumor agents.29 In addition, urea moiety consists in PI3K and
mTOR dual inhibitors PKI-179,30 PKI-58731 and PKI-402,32 which were developed by
Wyeth in clinical trial for the treatment of solid tumors.
Tertiary amine group is considered as a water soluble moiety in drug discovery.
N-methylpiperazinyl, pyrrolidyl and diethylamino can be found in drug structure.33
What's more, the pharmacokinetic property and water solubility of a drug could be
significantly improved after a tertiary amine moiety was attached.34
Given that tertiary amine and urea are important moieties in drug design, we
intend to combine tertiary amine group, urea moiety and 4-anilinoquinazoline scaffold
into one molecule to discover novel anticancer agents and EGFR-TK inhibitors
(Figure 2). Herein, we report the synthesis and biological activities evaluation of
thirty new compounds.
3

Figure 2. The design of target compounds
2. Results and discussion
2.1. Chemistry
The synthetic routes for the title compounds 1, 2 are outlined in Scheme1.
Scheme 1. Reagents and conditions: (a) (COCl)2, CH2Cl2, rt, 2 h; (b) secondary amine, THF,
0C to rt, 3 h; (c) 2 M NaOH, MeOH, rt, 4 h; (d) i: SOCl2, DMF, reflux, 2 h; ii: NaN3, THF/H2O,
DIPEA, rt, 4 h; (e) secondary amine, CH2Cl2, NaBH(OAc)3, rt, 5 h; (f) toluene, reflux, 2 h.
Commercially available 4-methoxycarbonylbenzoic acid was used as starting
material to prepare intermediates 5. The details were previously described in our
work.25 Intermediate 6 was prepared by reductive amination of methyl
4-formylbenzoate and secondary amine. The obtained 6 was hydrolyzed and
4

subsequently acidified to produce corresponding acid, which is an amino acid and
difficult to purify. So the crude product acid reacted with thionyl chloride to give acid
chloride, the latter reacted with sodium azide afforded acyl azide 7.
6-Amino-4-anilinoquinazolines were prepared according to the reported steps.35
Heating the acyl azide 5 in toluene produced isocyanate, which was not isolated and
reacted with 6-amino-4-anilinoquinazoline to produce targeted compounds 1a-1e.
Alternatively, refluxing the mixture of acyl azide 7, 6-amino-4-anilinoquinazolines
and toluene yielded compounds 2a-2o. Also, compound 2p was prepared by the
refluxing of 7 and (R)-6-amino-4-(1-phenylethylamino)quinazoline in toluene.
To probe the structure-activity relationship (SAR) of compounds with a
tertiary-amino substituted group at the m-position of phenyl urea moiety, compound
2q and 2r were designed and synthesized (Scheme 2). Intermediate 9 was prepared
from 3-nitrobenzaldehyde according to the reported steps.36 4-(3-Trifluoromethyl
phenylamino)quinoline-6-amine successively reacted with carbonyldimidazole (CDI)
and 9 to produce compound 2q and 2r.
Scheme 2. Reagents and conditions: (a) piperidine or pyrrolidine, CH2Cl2, NaBH(OAc)3, rt, 5 h;
(b) iron powder, NH4Cl, MeOH/H2O, reflux, 1 h; (c) i: CDI, MeCN, rt, 8 h; ii: 9, reflux, 4 h.
To further discuss the activity of the compounds with a substituted group at the
p-position of urea moiety, compounds 3a-3c were synthesized according to the same
steps in the preparation of 2a. The nucleophilic substitution of a secondary amine with
p-fluoronitrobenzene yielded compound 10, which was reduced by catalytic
hydrogenation to produce amine 11. The successive reaction of
5

4-(3-trifluoromethylphenylamino)quinozoline-6-amine with CDI and 11 produced
compounds 3d and 3e (Scheme 3).
Scheme 3. Reagents and conditions: (a) toluene, reflux, 2 h. (b) secondary amine, DMF, K2CO3,
50 C, 1 h; (c) Pd-C, H2, EtOH, rt, 4 h; (d) i: CDI, acetonitrile, rt, 8 h; ii: 11, reflux, 4 h.
To boost the structural diversity of the design compounds, the 6-position of
4-anilinoquinazoline was combined with 2-(4-methylpiperazin-1-yl)ethyl urea to yield
compounds 4. The 4-(3-trifluoromethylphenylamino) quinolinyl-6-amine successively
reacted with CDI and 2-(4-methylpiperazin-1-yl)ethylamine to produce compound 4a.
Likewise, (R)-4-(1-phenylethylamino) quinazolinyl-6-amine was used as starting
material to prepared 4b. The synthetic route is depicted in Scheme 3.
Scheme 4. Reagents and conditions: (a) i: CDI, acetonitrile, rt, 8 h; ii: 2-(4-methylpiperazin-
1-yl)ethylamine, rt, to reflux, 4 h.
1
All the newly synthesized compounds were characterized by H-NMR and
HRMS.
2.2. The antiproliferative activity in vitro
6

The antiproliferative activities of synthesized compounds were evaluated against
human epithelial carcinoma cell line (A431) and lung adenocarcinoma epithelial cell
line (A549) by applying the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-2H-tetrazolium
bromide (MTT) colorimetric assay. The EGFR-TK inhibitor gefitinib was used as the
positive control. The results are summarized in Table 1.
Table 1 Antiproliferative activities of compounds against two cancer cell lines (
, n = 3)
x s
IC50 (μM)
compds
R1
R2
A431
A549
1a
1b
1c
1d
1e
2a
2b
2c
2d
2e
2f
CH3N(CH2CH2)2N-
CH3N(CH2CH2)2N-
CH3N(CH2CH2)2N-
CH3N(CH2CH2)2N-
O(CH2CH2)2N-
CH3N(CH2CH2)2N-
CH3N(CH2CH2)2N-
CH3N(CH2CH2)2N-
CH3N(CH2CH2)2N-
CH3N(CH2CH2)2N-
CH3N(CH2CH2)2N-
CH3N(CH2CH2)2N-
CH3N(CH2CH2)2N-
(CH2)5N-
3-Cl-4-F
3-CCH
3-CF3
>10
>10
>10
>10
>10
8.31±1.21
7.39±0.52
>10
3-Cl-4-(3-FBnO)
3-Cl-4-F
3-Cl-4-F
3-CCH
3,4-diF
>10
>10
4.00±0.20
2.81±0.05
>10
3.48±0.12
5.17±0.70
>10
3-Cl-4-(3-FBnO)
3-Cl-4-OCH3
3-CF3
0.64±0.04
2.01±0.10
2.41±0.19
5.63±1.01
>10
0.90±0.15
3.57±0.33
1.65±0.09
>10
2g
2h
2i
3-CO2CH3
3-CON(CH2)4
3-CF3
>10
1.55±0.14
1.36±0.04
8.49±0.78
3.98±0.39
1.03±0.20
>10
1.05±0.07
0.83±0.11
4.03±0.43
2.07±0.20
1.12±0.20
>10
2j
(CH2)4N-
3-CF3
2k
2l
O(CH2CH2)2N-
(CH3CH2)2N-
3-CF3
3-CF3
2m
2n
2o
2p
2q
2r
3a
(CH3)2N-
3-CF3
O(CH2CH2)2N-
(CH3CH2)2N-
3-CCH
3-CCH
1.24±0.17
>10
2.25±0.10
>10
1.33±0.09
1.23±0.08
>10
1.06±0.19
0.86±0.20
>10
H-
7

3b
3c
CH3-
>10
4.14±0.58
9.47±1.40
3.52±0.39
2.76±0.12
>10
CH3O-
4.95±0.85
6.46±0.77
2.20±0.23
>10
3d
(CH3CH2)2N-
CH3N(CH2CH2)2N-
3e
4a
4b
>10
>10
gefitinib
3.74±0.41
5.76±0.42
As the data listed in table 1, most designed compounds displayed significant
antiproliferative activities against two human cancer cell lines, some of which were
more potent than that of gefitinib. The amine moiety was combined by a benzoyl
group in compounds 1, which showed poor antiproliferative activities. While in the
structure of compounds 2, the tertiary amine moiety was attached by a benzyl group.
The antiproliferative activity of compounds 2 is higher than that of compounds 1
against the two cancer cell lines. In order to discover compounds with potent
antiproliferative activity and explore structure-activity relationship of compounds 2,
we initially focused on the variation of substituents at 4-phenylamino group attached
to the 4-position of quinazoline. Except compounds 2c, 2g and 2h, compounds 2a, 2b,
2d, 2e and 2f displayed potent antiproliferative activities against the two cancer cell
lines. When 4-phenylamino group was replaced by the 1-phenylethylamino group
(compound 2p), the activity dropped dramatically. Compound 2d displayed
significant antiproliferative activities against the two cancer cell lines and its activity
against A431 is 5.8-fold to gefitinib. Changing 3-fluorobenzyloxy group in 2d to
methoxy (compound 2e) resulted in a decrease in activity. Compound 2f with a
3-trifluoromethyl group also showed potent antiproliferative activities against the two
cancer cell lines. These results suggest that the activity of compounds 2 is closely
related to the substituted group at the 4-position of quinazoline. Taking molecular
weight into consideration, we chose compound 2f to carry on the further SAR probe.
Next the 3-trifluoromethyl group was remained, and the N-methylpiperazin-1-yl
group was replaced by tertiary amino substituents, such as piperidyl, pyrrolidinyl,
4-morpholinyl, diethylamino, and dimethylamino to prepare compounds 2i-2m. The
8

data in Table 1 showed that these five compounds displayed potent antiproliferative
activities against the two cancer cell lines. Moreover, the activity of compound 2j
increased 3 to 5-fold to gefitinib in cell-based activity. The fact that the activity of
compounds 2k and 2n is weaker than 2l and 2o indicates that morpholinyl moiety is
an unfavorable substituent group in design compound. These results may be related to
the poor cell permeability of morpholinyl moiety.37 After the tertiary amino moiety
was moved from the p-position of phenyl urea moiety to the m-position, compound 2q
and 2r displayed the potent antiproliferative activities which is similar to that of
compound 2i and 2j.
To probe the effect of tertiary amino moiety on activity, compounds 3 were
synthesized. Without tertiary amino group, the activity of compounds 3a, 3b and 3c
against two cancer lines dropped significantly. Compounds 3d and 3e bearing an
amino substituents at the p-position of the phenylurea moiety exhibited an
improvement activity compared with compounds 3a, 3b and 3c, but a decline in
activity compared to compounds 2l and 2f. These results reveal that the tertiary amino
moiety attached by a benzyl group is favorable for the antiproliferative activity.
Furthermore, the replacement of phenylurea with 2-(4-methylpiperazin-1-yl)ethylurea
(compounds 4a, 4b) resulted in a total loss in antiproliferative activities. Building on
the discussions above, we can reveal a SAR in which diarylurea and tertiary amino
moiety are beneficial for the antiproliferative activities of gefitinib analogues. These
results accord with our design idea.
2.3. EGFR inhibitory activity assay
Table 2 The inhibitory activities of selected compounds against kinase
IC50 (nM)
compds
EGFRwt
8.4
EGFRL858R
VEGFR-2
>1000
nt
2d
2i
nt
10.5
11.5
2.6
nt
5.0
2j
4.1
565
gefitinib
sorafenib
1.3
nt
nt
1.2
nt = not tested.
9

Next, to elucidate the mechanism of antiproliferative activities of title
compounds, three compounds were selected to evaluate their inhibitory activity
against EGFR and VEGFR-2 by performing an ATP depletion assay.38 Gefitinib and
sorafenib were used as the positive drugs. The data are summarized in Table 2.
The data in Table 2 indicate that the selected compounds 2d, 2i and 2j display
remarkable potency against EGFRwt, compounds 2i and 2j exhibit potent inhibitory
activity against EGFR(L858R) and compound 2j displays weak activity against
VEGFR-2. These results suggest that the tested compounds are EGFR inhibitors.
We take into consideration both the antiproliferative activities and EGFR
inhibitory activities of compounds 2d, 2i, 2j and gefitinib together. Compounds 2d, 2i
and 2j exhibited more potent activities than gefitinib in cell based-activity, but weaker
activities than gefitinib in enzymatic-based activities. These results may relate to other
property of compounds 2d, 2i and 2j, such as pharmacokinetics.37
2.4. Anticancer effects in established nude mouse A549 xenograft model in vivo
Compound 2j displayed potent antiproliferative activity against A549 cell and
inhibitory activity against EGFR. Thus, we evaluated whether compound 2j could
inhibit cancer growth in an established nude mouce A549 xenograft model in vivo.
Gefitinib (50 mg/kg) was used as positive drug. 2j was dissolved and dosed orally at
50 mg/kg once a day for 12 days. The control group was administrated orally the
solvent only.
The change in the cancer volumes was depicted in Figure 3. In this model, the
growth of cancer in both treatment group and positive group can be inhibited
completely compared to the control group (P < 0.001). The curves in Figure 3 indicate
that compound 2j displays a little stronger anticancer effect than gefitinib. These
results suggest that compound 2j is an effective anticancer agent in vivo. In addition,
the significant weight loss of tested animals was not observed during the experimental
period.
10

Figure 3 The anticancer effect of compound 2j in established xenograft A549 lung
adenocarcinoma models (n = 5). Mice bearing cancers were orally administered solvent,
compound 2j and gefitinib once daily for 12 days. **P < 0.001 vs solvent.
2.5. Docking study
To further explain the potent activities of compound 2j and 2r, we performed a
docking analysis utilizing the C-DOCKER program within Discovery Studio 2.5
software package. Docking simulations were performed on the kinase domain of
EGFRwt (PDB code 1M17)39 and EGFRL858R (PDB code 4LQM)40. From the docking
results of compound 2j and 2r with EGFRwt (Figure 4, A and B), we observed that: (1)
both compounds 2j and 2r formed hydrogen bonds with hinge residues Met769 and
Gln767 by oxygen atom and hydrogen atom of the urea moiety; (2) the N1 of the
quinazoline moiety in compound 2j and 2r formed two hydrogen bonds with Thr830
and Asp831, and the N3 of the quinazoline moiety formed two hydrogen bonds with
Lys721.
From the docking results of compound 2j and 2r with EGFRL858R (Figure 4, C
and D), we observed that: (1) the two compounds formed two hydrogen bonds with
the hinge residue Met793 by the urea moiety; (2) the 4-amino moiety of compound 2j
formed hydrogen bond with Asp800, while the trifluoromethyl moiety of compound
2r formed two hydrogen bonds with Ser720.
11

Figure 4. Docking modes of compounds 2j and 2r with EGFRwt (A and B); compounds 2j and
2r with EGFRL858R (C and D). Selected residues Lys721, Gln767, Met769, Thr830, Asp831 in
EGFRwt and Ser720, Met793, Asp800 in EGFRL858R are shown. Green dashed lines indicate
hydrogen bonds.
3. Conclusion
In present study, a series of 4-anilinoquinazoline derivatives bearing diarylurea
and tertiary-amine moiety at its 6-position were synthesized and characterized. The
designed compounds displayed significant antiproliferative activities against A431
cell and A549 cell. The SAR of the tested compounds was discussed. The compounds
2i and 2j with potent antiproliferative activities exhibited potent inhibitory activity
against EGFRwt and EGFRL858R. Compound 2j can suppress the cancer growth in vivo
as well. These results suggest that 4-anilinoquinazoline derivatives bearing diarylurea
and tertiary amino moiety at its 6-position can serve as anticancer agents and EGFR
inhibitors.
4. Experimental
12

4.1. Chemistry
Unless specified otherwise, all starting materials, reagents and solvents were
commercially available. All reactions were monitored by thin-layer chromatography
on silica gel plates (GF-254) and visualized with UV light. All the melting points
were determined on a Beijing micro melting-point apparatus and thermometer was
uncorrected. 1H-NMR spectra were recorded on a 400 Bruker NMR spectrometer with
tetramethylsilane (TMS) as an internal reference. All chemical shifts are reported in
parts per million (ppm). High-resolution mass measurements were performed using
electrospray ionization (positive mode) on a quadrupole time-of-flight (QTOF) mass
spectrometer (Maxis Q-TOF, Bruker Inc.).
4.1.1. General procedures for the synthesis of compound 1a-1e
The intermediate 5 was prepared from 4-(methoxycarbonyl) benzoic acid as the
steps we previously reported.25 6-Amino-4-anilinoquinazoline was prepared as
reported approach.35
The solution of compound 5 (0.27 g, 0.99 mmol) in toluene (5.0 mL) was
refluxed for 1 h under N2 and cooled to about 30 C. 6-Amino-4-anilinoquinazoline
(0.99 mmol) was added. Then the mixture was stirred at 30C for another 2 h. The
solvent was removed and the residue was purified by flash column chromatograph
over silica gel (chloroform/methanol = 10:1, v/v) to give 1a–1e as white solids.
4.1.2.
1-(4-((3-Chloro-4-fluorophenyl)amino)quinazolin-6-yl)-3-(4-(4-methyl
piperazin-1-carbonyl)phenyl)urea (1a)
1
Yield 45.4%. mp: 183.5-184.5 C. H-NMR (DMSO-d6): δ 9.89 (s, 1H, N-H),
9.13 (s, 1H, N-H), 9.07 (s, 1H, N-H), 8.55 (s, 1H, Ar-H), 8.51 (d, 1H, Ar-H), 8.04 (s,
1H, Ar-H), 7.90 (d, 2H, Ar-H, J = 8.8 Hz), 7.78 (d, 1H, Ar-H), 7.58 (d, 2H, Ar-H, J =
8.4 Hz), 7.40 (t, 1H, Ar-H), 7.37 (s, 1H, Ar-H), 7.35 (s, 1H, Ar-H), 7.22 (d, 1H, Ar-H),
4.22 (s, 1H, C≡CH), 3.48 (br, 4H, 2×N-CH2), 2.32 (br, 4H, 2×N-CH2), 2.20 (s, 3H,
N-CH3). 13C-NMR (DMSO-d6): δ 169.4, 157.5, 153.7 (d, JC-F = 241 Hz), 153.0, 152.9,
146.4, 141.3, 138.0, 137.2, 129.5, 129.0, 128.7 (2×CH), 127.1, 124.1, 123.0, 119.2,
118.1 (2×CH), 116.9, 116.0, 110.6, 54.9, 46.0 (4×CH2). ESI-HRMS m/z: calc’d for
C27H25ClFN7O2 [M+H]+: 534.1821; found: 534.1817.
4.1.3.
1-(4-((3-Ethynylphenyl)amino)quinazolin-6-yl)-3-(4-(4-methylpiperazin-
13

1-carbonyl)phenyl)urea (1b)
1
Yield 42.5%. mp: 168.0-168.5 C. H-NMR (DMSO-d6): δ 9.83 (s, 1H, N-H),
9.13 (s, 1H, N-H), 9.05 (s, 1H, N-H), 8.55 (s, 1H, Ar-H), 8.51 (d, 1H, Ar-H), 8.04 (s,
1H, Ar-H), 7.90 (d, 2H, Ar-H, J = 8.8 Hz), 7.78 (d, 1H, Ar-H), 7.58 (d, 2H, Ar-H, J =
8.4 Hz), 7.40 (t, 1H, Ar-H), 7.37 (s, 1H, Ar-H), 7.35 (s, 1H, Ar-H), 7.22 (d, 1H, Ar-H),
4.22 (s, 1H, C≡CH), 3.48 (br, 4H, 2×N-CH2), 2.32 (br, 4H, 2×N-CH2), 2.20 (s, 3H,
N-CH3). 13C-NMR (DMSO-d6): δ 169.4, 157.5, 153.1, 152.9, 146.3, 140.9, 139.2,
138.0, 129.6, 129.4, 129.0, 128.8 (2×CH), 127.1, 126.7, 125.4, 123.0, 122.1, 118.3
(2×CH), 116.1, 110.2, 84.1, 81.0, 54.6, 48.0 (2×CH2), 45.4 (2×CH2). ESI-HRMS m/z:
calc’d for C29H27N7O2 [M+H]+: 506.2304; found: 506.2301.
4.1.4.
1-(4-((3-Trifluoromethylphenyl)amino)quinazolin-6-yl)-3-(4-(4-methyl
piperazin-1-carbonyl)phenyl)urea (1c)
Yield 54.3%. mp: 190.0~192.0 C . 1H-NMR (DMSO-d6): δ 10.03 (s, 1H, N-H),
9.19 (s, 1H, N-H), 9.15 (s, 1H, N-H), 8.58 (s, 2H, Ar-H), 8.30 (s, 1H, Ar-H), 8.22 (d,
1H, Ar-H, J = 8.4 Hz), 7.89 (d, 1H, Ar-H, J = 9.2 Hz), 7.79 (d, 1H, Ar-H, J = 9.2 Hz),
7.63 (t, 1H, Ar-H), 7.59 (d, 2H, Ar-H, J = 8.8 Hz), 7.45 (d, 1H, Ar-H, J = 7.6 Hz),
7.36 (d, 2H, Ar-H, J = 8.8 Hz), 3.48 (br, 4H, 2×N-CH2), 2.31 (br, 4H, 2×N-CH2), 2.20
(s, 3H, N-CH3). 13C-NMR (DMSO-d6): δ 169.4, 157.5, 153.2, 152.9, 146.4, 141.0,
138.6, 138.2, 130.3, 130.0, 129.5, 129.1, 128.8 (2×CH), 127.1, 126.1, 124.7 (d, JC-F
271 Hz), 119.9, 118.5, 118.0 (2×CH), 116.1, 110.2, 55.6, 48.4 (4×CH2). ESI-HRMS
m/z: calc’d for C28H26F3N7O2 [M+H]+: 550.2178; found: 550.2173.
=
4.1.5.
1-(4-((3-Chloro-4-(3-fluorophenylmethoxy)phenyl)amino)quinazolin-
6-yl)-3-(4-(4-methylpiperazin-1-carbonyl)phenyl)urea (1d)
1
Yield 34.7%. mp: 166.0~167.5 C . H-NMR (DMSO-d6): δ 9.78 (s, 1H, N-H),
9.14 (s, 1H, N-H), 9.05 (s, 1H, N-H), 8.50 (s, 1H, Ar-H), 8.48 (d, 1H, Ar-H), 8.00 (d,
1H, Ar-H), 7.87 (d, 1H, Ar-H, J = 9.2 Hz), 7.75 (d, 1H, Ar-H, J = 8.8 Hz), 7.71 (d, 1H,
Ar-H, J = 9.2 Hz), 7.57 (d, 2H, Ar-H), 7.44-7.51 (m, 1H, Ar-H), 7.31-7.37 (m, 4H,
Ar-H), 7.26 (d, 1H, Ar-H, J = 8.8 Hz), 7.20 (m, 1H, Ar-H), 5.26 (s, 2H, ArCH2-O),
3.44 (br, 4H, 2×N-CH2), 2.33 (br, 4H, 2×N-CH2), 2.20 (s, 3H, N-CH3). 13C-NMR
(DMSO-d6): δ 169.4, 162.7 (d, JC-F = 242 Hz), 157.6, 153.1, 153.0, 150.2, 146.4,
14

140.1, 140.0, 138.8, 138.2, 132.2, 131.2, 130.4, 128.9, 128.6 (2×CH), 126.9, 126.8,
124.5, 123.8, 123.0, 121.6, 118.9, 118.0 (2×CH), 116.0, 110.4, 56.0, 48.2 (4×CH2).
ESI-HRMS m/z: calc’d for C34H31ClFN7O3 [M+H]+: 640.2239; found: 640.2237.
4.1.6.
1-(4-((3-Chloro-4-fluorophenyl)amino)quinazolin-6-yl)-3-(4-morpholine-
4-carbonyl)phenyl)urea (1e)
1
Yield 37.4%. mp: 173.0~174.5 C . H-NMR (DMSO-d6): δ 9.92 (s, 1H, N-H),
9.15 (s, 1H, Ar-H), 9.08 (s, 1H, N-H), 8.55 (s, 2H, Ar-H), 8.52 (s, 1H, Ar-H), 8.17 (d,
1H, Ar-H), 7.88 (d, 1H, Ar-H), 7.77-7.83 (m, 2H, Ar-H), 7.59 (d, 2H, Ar-H, J = 8.0
Hz), 7.45 (t, 1H, Ar-H), 7.40 (d, 2H, Ar-H, J = 8.0 Hz), 3.60 (br, 4H, 2×O-CH2), 3.51
13
(br, 4H, 2×N-CH2). C-NMR (DMSO-d6): δ 169.4, 157.5, 154.1 (d, JC-F = 241 Hz),
153.2, 152.9, 146.4, 139.0, 138.3, 137.8, 129.5, 129.1, 128.8 (2×CH), 127.0, 124.0,
123.2, 119.8, 118.2 (2×CH), 116.8, 116.0, 110.4, 56.6 (2×CH2), 50.8 (2×CH2).
ESI-HRMS m/z: calc’d for C26H22ClFN6O3 [M+H]+: 521.1504; found: 521.1528.
4.1.7. General procedures for the synthesis of compounds 6a-6f
Compounds 6a-6f were prepared according to the reported method.41
4.1.8. Methyl 4-((4-methylpiperazin-1-yl)methyl)benzoate (6a)
1
Yield 86.5%. H-NMR (CDCl3): δ 8.00 (d, 2H, Ar-H, J = 8.4 Hz), 7.42 (d, 2H,
Ar-H, J = 8.4 Hz), 3.92 (s, 3H, OCH3), 3.57 (s, 2H, Ar-CH2-N), 2.49 (br, 8H,
4×N-CH2), 2.31 (s, 3H, N-CH3).
4.1.9. Methyl 4-(morpholinomethyl)benzoate (6b)
1
Yield 85.0%. H-NMR (CDCl3): δ 8.00 (d, 2H, Ar-H, J = 8.0 Hz), 7.42 (d, 2H,
Ar-H, J = 8.4 Hz), 3.92 (s, 3H, OCH3), 3.72 (t, 4H, 2×O-CH2, J = 4.4 Hz), 3.55 (s, 2H,
Ar-CH2-N), 2.45 (t, 4H, 2×N-CH2, J = 4.4 Hz).
4.1.10. Methyl 4-((diethylamino)methyl)benzoate (6c)
1
Yield 83.3%. H-NMR (CDCl3): δ 7.99 (d, 2H, Ar-H, J = 8.0 Hz), 7.43 (d, 2H,
Ar-H, J = 8.0 Hz), 3.91 (s, 3H, OCH3), 3.63 (s, 2H, Ar-CH2-N), 2.54 (q, 4H, 2×N-CH2,
J = 7.2 Hz), 1.06 (t, 6H, 2×CH3, J = 7.2 Hz).
4.1.11. Methyl 4-(piperidin-1-ylmethyl)benzoate (6d)
1
Yield 90.2%. H-NMR (CDCl3): δ 8.00 (d, 2H, Ar-H, J = 8.4 Hz), 7.42 (d, 2H,
Ar-H, J = 8.4 Hz), 3.94 (s, 3H, OCH3), 3.54 (s, 2H, Ar-CH2-N), 2.40 (br, 4H,
15

2×N-CH2), 1.57-1.63 (m, 4H, 2×CH2), 1.41-1.49 (m, 2H, CH2).
4.1.12. Methyl 4-(pyrrolidin-1-ylmethyl)benzoate (6e)
1
Yield 87.3%. H-NMR (CDCl3): δ 8.08 (d, 2H, Ar-H, J = 8.4 Hz), 7.66 (d, 2H,
Ar-H, J = 8.4 Hz), 4.10 (s, 2H, Ar-CH2-N), 3.94 (s, 3H, OCH3), 3.04 (br, 4H,
2×N-CH2), 2.02-2.09 (m, 4H, 2×CH2).
4.1.13. Methyl 4-((dimethylamino)methyl)benzoate (6f)
1
Yield 86.4%. H-NMR (CDCl3): δ 8.06 (d, 2H, Ar-H, J = 8.0 Hz), 7.54 (d, 2H,
Ar-H, J = 8.0 Hz), 3.94 (s, 3H, OCH3), 3.76 (s, 2H, Ar-CH2-N), 2.46 (s, 6H,
2×N-CH3).
4.1.14. General procedures for the synthesis of compounds 7a-7f
The mixture containing compound 6 (2.51 mmol), methanol (5.0 mL), 2 M
NaOH (1.6 mL) was stirred at room temperature for 4 h, concentrated under reduced
pressure and acidified with concentrated hydrochloric acid to pH 2. The aqueous
phase was concentrated to dryness under reduced pressure to afford the crude
4-aminomethylbenzoic acid (containing sodium chloride) which was carried on the
next step without further purification. The 4-aminomethylbenzoic acid (2.51 mmol)
and 2 drops of DMF were added to thionyl chloride (10 mL). The mixture was
refluxed for 2 h. The volatile was removed under reduced pressure to give a pale
yellow solid which was then dissolved in anhydrous THF (10 mL) and was added
dropwise to a suspension of sodium azide (0.24 g, 3.77 mmol) in 10 mL of THF/H2O
(4:1, v/v) at 0-5 C. Then the mixture was stirred at room temperature overnight and
THF was removed under reduced pressure. The aqueous phase was extracted with
dichloromethane (20 mL  3). The organic layer was combined, washed with brine
(20 mL), dried (Na2SO4), filtered and concentrated under reduced pressure to produce
compounds 7a-7f as yellow or brown oil.
4.1.15. 4-((4-Methylpiperazin-1-yl)methyl)benzoyl azide (7a)
1
Yield 76.3%. H-NMR (CDCl3): δ 7.99 (d, 2H, Ar-H, J = 8.0 Hz), 7.44 (d, 2H,
Ar-H, J = 8.0 Hz), 3.58 (s, 2H, Ar-CH2-N), 2.48 (br, 8H, 4×N-CH2), 2.31 (s, 3H,
N-CH3). MS: 260 [M+H]+.
4.1.16. 4-(Morpholinomethyl)benzoyl azide (7b)
16

1
Yield 81.0%. H-NMR (CDCl3): δ 8.00 (d, 2H, Ar-H, J = 8.4 Hz), 7.46 (d, 2H,
Ar-H, J = 8.4 Hz), 3.74 (t, 4H, 2×O-CH2, J = 4.4 Hz), 3.58 (s, 2H, Ar-CH2-N), 2.47 (t,
4H, 2×N-CH2, J = 4.0 Hz). MS: 247 [M+H]+.
4.1.17. 4-((Diethylamino)methyl)benzoyl azide (7c)
Yield 70.5%. 1H-NMR (CDCl3): δ 8.06 (br, 2H, Ar-H), 7.60 (br, 2H, Ar-H), 2.93
(s, 2H, Ar-CH2-N), 1.70 (br, 4H, 2×N-CH2), 1.28 (br, 6H, 2×CH3). MS: 232 [M+H]+.
4.1.18. 4-(Piperidin-1-ylmethyl)benzoyl azide (7d)
1
Yield 69.3%. H-NMR (CDCl3): δ 7.98 (d, 2H, Ar-H, J = 8.0 Hz), 7.46 (d, 2H,
Ar-H, J = 8.0 Hz), 3.59 (s, 2H, Ar-CH2-N), 2.44 (br, 4H, 2×N-CH2), 1.65-1.59 (m, 4H,
2×CH2), 1.53-1.56 (m, 2H, CH2). MS: 244 [M+H]+.
4.1.19. 4-(Pyrrolidin-1-ylmethyl)benzoyl azide (7e)
1
Yield 78.5%. H-NMR (CDCl3): δ 7.90 (d, 2H, Ar-H, J = 8.0 Hz), 7.43 (d, 2H,
Ar-H, J = 8.0 Hz), 3.68 (s, 2H, Ar-CH2-N), 2.52 (br, 4H, 2×N-CH2), 1.79 (br, 4H,
2×CH2). MS: 230 [M+H]+.
4.1.20. 4-((Dimethylamino)methyl)benzoyl azide (7f)
1
Yield 76.4%. H-NMR (CDCl3): δ 8.03 (d, 2H, Ar-H, J = 8.0 Hz), 7.51 (d, 2H,
Ar-H, J = 8.0 Hz), 3.65 (s, 2H, Ar-CH2-N), 2.37 (s, 6H, 2×N-CH3). MS: 204 [M+H]+.
4.1.21. General procedures for the synthesis of compounds 2a-2p and 3a-3c
Compound 7 or benzoyl azide or 4-methylbenzoyl azide (0.77 mmol) was added
to toluene (5 mL). The mixture was stirred, added 4-anilinoquinazoline-6-amine or
(R)-6-amino-4-(1-phenylethylamino)quinazoline (0.62 mmol), refluxed for 2 h,
cooled to room temperature. The precipitate was filtered and purified by column
chromatography on silica gel (dichloromethane /methanol = 30:1, v/v) to produce
compounds 2a-2p or 3a-3c as yellow or off-white powder.
4.1.22.
1-(4-((3-Chloro-4-fluorophenyl)amino)quinazolin-6-yl)-3-(4-((4-methyl
piperazin-1-yl)methyl)phenyl)urea (2a)
1
Yield 68.5%. mp: 191.5-192.0 C. H-NMR (DMSO-d6): δ 9.90 (s, 1H, N-H),
9.02 (s, 1H, N-H), 8.92 (s, 1H, N-H), 8.54 (s, 1H, Ar-H), 8.50 (s, 1H, Ar-H), 8.16-8.19
(m, 1H, Ar-H), 7.88 (dd, 1H, Ar-H, J1 = 1.6 Hz, J2 = 8.8 Hz), 7.80-7.85 (m, 1H, Ar-H),
7.77 (d, 1H, Ar-H, J = 8.8 Hz), 7.47 (s, 1H, Ar-H), 7.44 (d, 2H, Ar-H, J = 8.0 Hz),
17

7.21 (d, 2H, Ar-H, J = 8.0 Hz), 3.46 (s, 2H, Ar-CH2-N), 2.34 (br, 8H, 4×N-CH2), 2.17
(s, 3H, N-CH3). 13C-NMR (DMSO-d6): δ 157.5, 153.7 (d, JC-F = 241 Hz), 153.1, 152.9,
146.2, 138.8, 138.3, 137.2, 132.2, 129.9 (2×CH), 129.0, 127.0, 124.1, 123.0, 119.2,
118.6 (2×CH), 116.9, 116.0, 110.2, 62.0, 55.1 (2×CH2), 52.7 (2×CH2), 46.0.
ESI-HRMS m/z: calc’d for C27H28ClFN7O [M+H]+: 520.2028; found: 520.2022.
4.1.23.
1-(4-((3-Ethynylphenyl)amino)quinazolin-6-yl)-3-(4-((4-methyl
piperazin-1-yl)methyl)phenyl)urea (2b)
1
Yield 64.5%. mp: 129.5-131.0 C. H-NMR (DMSO-d6): δ 9.86 (s, 1H, N-H),
9.32 (s, 1H, N-H), 9.28 (s, 1H, N-H), 8.54 (s, 1H, Ar-H), 8.51 (s, 1H, Ar-H), 8.04 (s,
1H, Ar-H), 7.90 (d, 2H, Ar-H, J = 8.4 Hz), 7.76 (d, 1H, Ar-H, J = 8.0 Hz), 7.61 (d, 2H,
Ar-H, J = 8.4 Hz), 7.40 (t, 1H, Ar-H, J = 8.0 Hz), 7.20-7.25 (m, 3H, Ar-H), 4.23 (s,
1H, C≡CH), 3.46 (s, 2H, Ar-CH2-N), 3.01-3.09 (m, 4H, 2×N-CH2), 2.65 (s, 3H,
N-CH3), 2.37 (s, 4H, 2×N-CH2). 13C-NMR (DMSO-d6): δ 157.6, 153.2, 152.9, 146.2,
140.3, 139.2, 138.4, 131.3, 130.0 (2×CH), 129.3, 128.8, 127.0, 126.8, 125.4, 123.3,
122.1, 118.4 (2×CH), 116.2, 110.2, 84.1, 81.0, 61.9, 54.2 (2×CH2), 51.6 (2×CH2),
44.8. ESI-HRMS m/z: calc’d for C29H30N7O [M+H]+: 492.2512; found: 492.2506.
4.1.24.
1-(4-((3,4-Difluorophenyl)amino)quinazolin-6-yl)-3-(4-((4-methyl
piperazin-1-yl)methyl)phenyl)urea (2c)
1
Yield 42.3%. mp: 190.6-192.0 C. H-NMR (DMSO-d6): δ 9.90 (s, 1H, N-H),
9.02 (s, 1H, N-H), 8.92 (s, 1H, N-H), 8.55 (s, 1H, Ar-H), 8.50 (s, 1H, Ar-H),
8.05-8.12 (m, 1H, Ar-H), 7.89 (dd, 1H, Ar-H, J1 = 1.6 Hz, J2 = 8.8 Hz), 7.78 (d, 1H,
Ar-H, J = 8.8 Hz), 7.64 (d, 1H, Ar-H, J = 8.4 Hz), 7.41-7.50 (m, 3H, Ar-H), 7.21 (d,
2H, Ar-H, J = 8.0 Hz), 3.43 (s, 2H, Ar-CH2-N), 2.34 (br, 8H, 4×N-CH2), 2.16 (s, 3H,
N-CH3). 13C-NMR (DMSO-d6): δ 157.5, 153.1, 152.9, 149.2 (d, JC-F = 228 Hz),
146.3 (d, JC-F = 228 Hz), 146.2, 138.8, 138.3, 137.1, 132.2, 129.9 (2×CH), 129.0,
127.0, 118.9, 118.6 (2×CH), 117.4, 116.0, 111.6, 110.3, 62.1, 55.1 (2×CH2), 52.8
(2×CH2), 46.1. ESI-HRMS m/z: calc’d for C27H28F2N7O [M+H]+: 504.2323; found:
504.2329.
4.1.25. 1-(4-((3-Chloro-4-((3-fluorobenzyl)oxy)phenyl)amino)quinazolin-6-yl)-
3-(4-((4-methylpiperazin-1-yl)methyl)phenyl)urea (2d)
18

1
Yield 54.6%. mp: 197.3-198.7 C. H-NMR (DMSO-d6): δ 9.76 (s, 1H, N-H),
8.97 (s, 1H, N-H), 8.90 (s, 1H, N-H), 8.49 (s, 1H, Ar-H), 8.46 (d, 1H, Ar-H, J = 2.0
Hz), 8.01 (d, 1H, Ar-H, J = 2.0 Hz), 7.87 (dd, 1H, Ar-H, J1 = 2.0 Hz, J2 = 9.2 Hz),
7.75 (d, 1H, Ar-H, J = 9.2 Hz), 7.71 (d, 1H, Ar-H, J = 2.0 Hz), 7.44-7.52 (m, 3H,
Ar-H), 7.30-7.36 (m, 2H, Ar-H), 7.26 (d, 1H, Ar-H, J = 8.8 Hz), 7.21 (d, 2H, Ar-H, J
= 8.8 Hz), 7.15-7.19 (m, 1H, Ar-H), 5.26 (s, 2H, Ar-CH2-O), 3.39 (s, 2H, Ar-CH2-N),
2.33 (br, 8H, 4×N-CH2), 2.15 (s, 3H, N-CH3). 13C-NMR (DMSO-d6): δ 162.7 (d, JC-F
= 242 Hz), 157.6, 153.1, 153.0, 150.0, 146.2, 140.2, 140.1, 138.8, 138.1, 134.0, 132.2,
131.0, 129.8 (2×CH), 128.9, 126.8, 124.6, 123.8, 122.8, 121.4, 118.6 (2×CH), 116.0,
115.2, 114.5, 110.4, 69.8, 62.1, 55.1 (2×CH2), 52.8 (2×CH2), 46.1. ESI-HRMS m/z:
calc’d for C34H34ClFN7O2 [M+H]+: 626.2447; found: 626.2441.
4.1.26.
1-(4-((3-Chloro-4-methoxyphenyl)amino)quinazolin-6-yl)-3-(4-((4-
methylpiperazin-1-yl)methyl)phenyl)urea (2e)
1
Yield 61.2%. mp: 198.0-201.0 C. H-NMR (DMSO-d6): δ 9.74 (s, 1H, N-H),
8.98 (s, 1H, N-H), 8.91 (s, 1H, N-H), 8.49 (s, 1H, Ar-H), 8.46 (d, 1H, Ar-H, J = 2.0
Hz), 7.98 (d, 1H, Ar-H, J = 2.8 Hz), 7.87 (dd, 1H, Ar-H, J1 = 2.0 Hz, J2 = 8.8 Hz),
7.71-7.76 (m, 2H, Ar-H), 7.46 (d, 2H, Ar-H, J = 8.4 Hz), 7.18-7.23 (m, 3H, Ar-H),
3.88 (s, 3H, O-CH3), 3.39 (s, 2H, Ar-CH2-N), 2.35 (br, 8H, 4×N-CH2), 2.17 (s, 3H,
N-CH3). 13C-NMR (DMSO-d6): δ 157.6, 153.2, 153.1, 151.2, 146.1, 138.8, 138.1,
133.5, 132, 129.9 (2×CH), 128.9, 126.8, 124.6, 122.9, 120.7, 118.6 (2×CH), 116.0,
113.0, 110.4, 62.1, 56.7, 55.1 (2×CH2), 52.8 (2×CH2), 46.0. ESI-HRMS m/z: calc’d
for C28H31ClN7O2 [M+H]+: 532.2228; found: 532.2222.
4.1.27. 1-(4-((4-Methylpiperazin-1-yl)methyl)phenyl)-3-(4-((3-(trifluoromethyl)
phenyl)amino)quinazolin-6-yl)urea (2f)
1
Yield 23.8%. mp: 183.0-184.5 C. H-NMR (DMSO-d6): δ 10.02 (s, 1H, N-H),
9.04 (s, 1H, N-H), 8.93 (s, 1H, N-H), 8.57 (s, 1H, Ar-H), 8.55 (d, 1H, Ar-H, J = 2.0
Hz), 8.31 (s, 1H, Ar-H), 8.23 (d, 1H, Ar-H, J = 8.0 Hz), 7.89 (dd, 1H, Ar-H, J1 = 2.0
Hz, J2 = 8.8 Hz), 7.79 (d, 1H, Ar-H, J = 8.8 Hz), 7.63 (t, 1H, Ar-H, J = 8.0 Hz),
7.44-7.48 (m, 3H, Ar-H), 7.22 (d, 2H, Ar-H, J = 8.4 Hz), 3.39 (s, 2H, Ar-CH2-N),
2.35 (br, 8H, 4×N-CH2), 2.17 (s, 3H, N-CH3). 13C-NMR (DMSO-d6): δ 157.5, 153.1,
19

152.9, 146.3, 140.9, 138.8, 138.4, 132.2, 130.0, 129.9 (2×CH), 129.8, 129.0, 127.1,
126.1, 124.7 (d, JC-F = 266 Hz), 119.9, 118.7 (2×CH), 118.5, 116.1, 110.3, 62.0, 55.0
(2×CH2), 52.7 (2×CH2), 46.0. ESI-HRMS m/z: calc’d for C28H29F3N7O [M+H]+:
536.2386; found: 536.2380.
4.1.28.
Methyl 3-((6-(3-(4-((4-methylpiperazin-1-yl)methyl)phenyl)ureido)
quinazolin-4-yl)amino)benzoate (2g)
1
Yield 22.5%. mp: 144.5-147.0 C. H-NMR (DMSO-d6): δ 10.07 (s, 1H, N-H),
9.81 (s, 1H, N-H), 9.52 (s, 1H, N-H), 8.56 (s, 1H, Ar-H), 8.55 (s, 1H, Ar-H), 8.46 (s,
1H, Ar-H), 8.23 (d, 1H, Ar-H, J = 7.6 Hz), 7.92 (d, 1H, Ar-H, J = 8.4 Hz), 7.78 (d, 1H,
Ar-H, J = 8.8 Hz), 7.71 (d, 1H, Ar-H, J = 7.6 Hz), 7.50-7.59 (m, 3H, Ar-H), 7.26 (br,
2H, Ar-H), 3.89 (s, 3H, O-CH3), 3.51 (s, 2H, Ar-CH2-N), 2.95 (br, 8H, 4×N-CH2),
2.72 (s, 3H, N-CH3). 13C-NMR (DMSO-d6): δ 166.6, 158.0, 153.2, 152.3, 144.2,
140.1, 139.6, 138.8, 130.6, 130.5, 130.4, 129.4 (2×CH), 127.7, 127.5, 127.2, 124.9,
123.6, 118.4 (2×CH), 116.0, 110.3, 60.6, 52.7 (2×CH2), 49.5, 45.9 (2×CH2), 42.8.
ESI-HRMS m/z: calc’d for C29H32N7O3 [M+H]+: 526.2567; found: 526.2561.
4.1.29.
1-(4-((4-Methylpiperazin-1-yl)methyl)phenyl)-3-(4-((3-(pyrrolidin-1-yl
carbonyl)phenyl)amino)quinazolin-6-yl)urea (2h)
1
Yield 20.6%. mp: 161.5-163.0 C. H-NMR (DMSO-d6): δ 9.87 (s, 1H, N-H),
9.04 (s, 1H, N-H), 8.98 (s, 1H, N-H), 8.53 (s, 1H, Ar-H), 8.51 (s, 1H, Ar-H), 8.04 (s,
1H, Ar-H), 7.95 (d, 1H, Ar-H, J = 8.4 Hz), 7.91 (d, 1H, Ar-H, J = 8.8 Hz), 7.77 (d, 1H,
Ar-H, J = 8.8 Hz), 7.48 (s, 1H, Ar-H), 7.45 (d, 2H, Ar-H, J = 6.0 Hz), 7.26 (s, 1H,
Ar-H), 7.22 (d, 2H, Ar-H, J = 8.4 Hz), 3.45-3.51 (m, 4H, 2×CH2), 3.40 (s, 2H,
Ar-CH2-N), 2.39 (br, 8H, 4×N-CH2), 2.26 (s, 3H, N-CH3), 1.81-1.93 (m, 4H, 2×CH2).
13C-NMR (DMSO-d6): δ 168.6, 157.6, 153.2, 153.0, 146.2, 139.8, 139.0, 138.3, 137.7,
131.7, 130.0 (2×CH), 128.9, 128.8, 126.9, 123.7, 122.4, 121.2, 118.6 (2×CH), 116.2,
110.4, 61.8, 54.7 (2×CH2), 52.2 (2×CH2), 49.5 (2×CH2), 46.4, 26.5 (2×CH2).
ESI-HRMS m/z: calc’d for C32H37N8O2 [M+H]+: 565.3039; found: 565.3034.
4.1.30.
1-(4-((Piperidin-1-yl)methyl)phenyl)-3-(4-((3-(trifluoromethyl)phenyl)
amino)quinazolin-6-yl)urea (2i)
1
Yield 20.3%. mp: 156.0-158.0 C. H-NMR (DMSO-d6): δ 10.02 (s, 1H, N-H),
20

9.06 (s, 1H, N-H), 8.97 (s, 1H, N-H), 8.58 (s, 1H, Ar-H), 8.55 (d, 1H, Ar-H, J = 1.6
Hz), 8.30 (s, 1H, Ar-H), 8.23 (d, 1H, Ar-H, J = 8.0 Hz), 7.90 (dd, 1H, Ar-H, J1 = 1.6
Hz, J2 = 8.8 Hz), 7.80 (d, 1H, Ar-H, J = 8.8 Hz), 7.63 (t, 1H, Ar-H, J = 8.0 Hz), 7.49
(d, 2H, Ar-H, J = 8.0 Hz), 7.46 (d, 1H, Ar-H, J = 8.0 Hz), 7.26 (d, 2H, Ar-H, J = 8.0
Hz), 3.56 (s, 2H, Ar-CH2-N), 2.45 (br, 4H, 2×N-CH2), 1.54 (br, 4H, 2×CH2), 1.42 (br,
13
2H, CH2). C-NMR (DMSO-d6): δ 157.5, 153.1, 152.9, 146.3, 140.9, 138.7, 138.4,
132.5, 130.0, 129.8 (2×CH), 129.5, 129.0, 127.1, 126.0, 124.7 (d, JC-F = 270 Hz),
119.9, 118.6 (2×CH), 118.4, 116.1, 110.3, 62.9, 54.2 (2×CH2), 26.0 (2×CH2), 24.5.
ESI-HRMS m/z: calc’d for C28H28F3N6O [M+H]+: 521.2277; found: 521.2271.
4.1.31.
1-(4-(Pyrrolidin-1-yl-methyl)phenyl)-3-(4-((3-(trifluoromethyl)phenyl)
amino)quinazolin-6-yl)urea (2j)
1
Yield 21.5%. mp: 176.1-179.2 C. H-NMR (DMSO-d6): δ 10.02 (s, 1H, N-H),
9.03 (s, 1H, N-H), 8.91 (s, 1H, N-H), 8.57 (s, 1H, Ar-H), 8.56 (d, 1H, Ar-H, J = 2.0
Hz), 8.31 (s, 1H, Ar-H), 8.23 (d, 1H, Ar-H, J = 8.0 Hz), 7.89 (dd, 1H, Ar-H, J1 = 2.0
Hz, J2 = 8.8 Hz), 7.79 (d, 1H, Ar-H, J = 9.2 Hz), 7.63 (t, 1H, Ar-H, J = 8.0 Hz),
7.40-7.50 (m, 3H, Ar-H), 7.24 (d, 2H, Ar-H, J = 8.0 Hz), 3.53 (s, 2H, Ar-CH2-N),
13
2.44 (br, 4H, 2×N-CH2), 1.65-1.75 (m, 4H, 2×CH2). C-NMR (DMSO-d6): δ 157.5,
153.1, 152.9, 146.3, 140.9, 138.6, 138.3, 133.5, 130.0, 129.5, 129.4 (2×CH), 129.0,
127.1, 126.0, 124.7 (d, JC-F = 270 Hz), 118.7 (2×CH), 119.9, 118.4, 116.1, 110.3, 59.6,
53.9 (2×CH2), 23.5 (2×CH2). ESI-HRMS m/z: calc’d for C27H26F3N6O [M+H]+:
507.2120; found: 507.2115.
4.1.32. 1-(4-(Morpholinomethyl)phenyl)-3-(4-((3-(trifluoromethyl)phenyl)
amino)quinazolin-6-yl)urea (2k)
1
Yield 44.6%. mp: 186.0-188.0C. H-NMR (DMSO-d6): δ 10.02 (s, 1H, N-H),
9.00 (s, 1H, N-H), 8.90 (s, 1H, N-H), 8.57 (s, 1H, Ar-H), 8.55 (d, 1H, Ar-H, J = 2.4
Hz), 8.30 (s, 1H, Ar-H), 8.23 (d, 1H, Ar-H, J = 8.0 Hz), 7.89 (dd, 1H, Ar-H, J1 = 2.4
Hz, J2 = 8.8 Hz), 7.79 (d, 1H, Ar-H, J = 8.8 Hz), 7.63 (t, 1H, Ar-H, J = 8.0 Hz),
7.43-7.50 (m, 3H, Ar-H), 7.24 (d, 2H, Ar-H, J = 8.4 Hz), 3.57 (t, 4H, 2×O-CH2, J =
13
4.0 Hz), 3.41 (s, 2H, Ar-CH2-N), 2.34 (br, 4H, 2×N-CH2). C-NMR (DMSO-d6): δ
157.5, 153.1, 152.9, 146.3, 140.9, 138.9, 138.3, 131.7, 130.0 (2×CH), 129.8, 129.5,
21

129.0, 127.1, 126.1, 124.7 (d, JC-F = 271 Hz), 119.9, 118.7 (2×CH), 118.5, 116.1,
110.3, 66.6 (2×CH2), 62.5 (CH2), 53.5 (2×CH2). ESI-HRMS m/z: calc’d for
C27H26F3N6O2 [M+H]+: 523.2069; found: 523.2064.
4.1.33.
1-(4-(Diethylaminomethyl)phenyl)-3-(4-((3-(trifluoromethyl)phenyl)
amino)quinazolin-6-yl)urea (2l)
1
Yield 25.4%. mp: 134.8-137.0 C. H-NMR (DMSO-d6): δ 10.03 (s, 1H, N-H),
9.17 (br, 2H, 2×N-H), 8.58 (s, 1H, Ar-H), 8.55 (d, 1H, Ar-H, J = 1.6 Hz), 8.30 (s, 1H,
Ar-H), 8.23 (d, 1H, Ar-H, J = 8.0 Hz), 7.90 (dd, 1H, Ar-H, J1 = 2.0 Hz, J2 = 8.8 Hz),
7.80 (d, 1H, Ar-H, J = 9.2 Hz), 7.63 (t, 1H, Ar-H, J = 8.0 Hz), 7.41-7.57 (m, 3H,
Ar-H), 7.22-7.39 (m, 2H, Ar-H), 3.53 (s, 2H, Ar-CH2-N), 2.56 (br, 4H, 2×N-CH2),
0.92-1.14 (m, 6H, 2×CH3). 13C-NMR (DMSO-d6): δ 157.5, 153.1, 152.9, 146.3, 140.9,
138.9, 138.3, 130.9, 130.0 (2×CH), 129.8, 129.5, 129.0, 127.1, 126.1, 124.7 (d, JC-F
=
271 Hz), 119.9, 118.6 (2×CH), 118.5, 116.1, 110.3, 56.0, 46.3 (2×CH2), 21.6 (2×CH3).
ESI-HRMS m/z: calc’d for C27H28F3N6O [M+H]+: 509.2277; found: 509.2271.
4.1.34. 1-(4-((Dimethylamino)methyl)phenyl)-3-(4-((3-(trifluoromethyl)
phenyl)amino)quinazolin-6-yl)urea (2m)
1
Yield 34.8%. mp: 152.3-154.0 C. H-NMR (DMSO-d6): δ 10.06 (s, 1H, N-H),
9.46 (s, 1H, N-H), 9.43 (s, 1H, N-H), 8.57 (s, 1H, Ar-H), 8.56 (s, 1H, Ar-H), 8.31 (s,
1H, Ar-H), 8.23 (d, 1H, Ar-H, J = 8.4 Hz), 7.91 (d, 1H, Ar-H, J = 8.8 Hz), 7.80 (d, 1H,
Ar-H, J = 8.8 Hz), 7.63 (t, 1H, Ar-H, J = 8.0 Hz), 7.56 (d, 2H, Ar-H, J = 8.0 Hz), 7.45
(d, 1H, Ar-H, J = 7.6 Hz), 7.36 (d, 2H, Ar-H, J = 8.0 Hz), 3.86 (s, 2H, Ar-CH2-N),
13
2.48 (s, 6H, 2×N-CH3). C-NMR (DMSO-d6): δ 157.6, 153.2, 152.9, 146.3, 140.9,
140.4, 138.3, 131.3, 130.0 (2×CH), 129.8, 129.5, 129.0, 127.1, 126.1, 124.7 (d, JC-F
=
271 Hz), 119.9, 118.5, 118.4 (2×CH), 116.2, 110.3, 61.1, 43.1 (2×CH3). ESI-HRMS
m/z: calc’d for C25H24F3N6O [M+H]+: 481.1964; found: 481.1958.
4.1.35.
1-(4-((3-Ethynylphenyl)amino)quinazolin-6-yl)-3-(4-(morpholino
methyl)phenyl)urea (2n)
1
Yield 46.6%. mp: 204.6-206.0 C. H-NMR (DMSO-d6): δ 9.83 (s, 1H, N-H),
9.02 (s, 1H, N-H), 8.93 (s, 1H, N-H), 8.55 (s, 1H, Ar-H), 8.50 (s, 1H, Ar-H), 8.05 (s,
1H, Ar-H), 7.92 (s, 1H, Ar-H), 7.90 (s, 1H, Ar-H), 7.77 (d, 1H, Ar-H, J = 8.8 Hz),
22

7.45 (d, 2H, Ar-H, J = 8.0 Hz), 7.40 (t, 1H, Ar-H, J = 8.0 Hz), 7.20-7.26 (m, 3H,
Ar-H), 4.22 (s, 1H, C≡CH), 3.55-3.59 (m, 4H, 2×O-CH2), 3.37 (s, 2H, Ar-CH2-N),
13
2.34 (br, 4H, 2×N-CH2). C-NMR (DMSO-d6): δ 157.6, 153.1, 153.0, 146.3, 140.3,
138.9, 138.2, 131.6, 130.0 (2×CH), 129.3, 128.9, 127.1, 127.0, 125.4, 123.3, 122.2,
118.6 (2×CH), 116.1, 110.4, 84.0, 81.0, 66.6 (2×CH2), 62.4, 53.5 (2×CH2).
ESI-HRMS m/z: calc’d for C28H27N6O2 [M+H]+: 479.2195; found: 479.2190.
4.1.36.
1-(4-((Diethylamino)methyl)phenyl)-3-(4-((3-ethynylphenyl)amino)
quinazolin-6-yl)urea (2o)
1
Yield 32.5%. mp: 181.0-182.0 C. H-NMR (DMSO-d6): δ 9.83 (s, 1H, N-H),
9.00 (s, 1H, N-H), 8.90 (s, 1H, N-H), 8.54 (s, 1H, Ar-H), 8.49 (s, 1H, Ar-H), 8.04 (s,
1H, Ar-H), 7.92 (s, 1H, Ar-H), 7.89 (s, 1H, Ar-H), 7.77 (d, 1H, Ar-H, J = 9.2 Hz),
7.46 (d, 2H, Ar-H, J = 8.0 Hz), 7.41 (t, 1H, Ar-H, J = 8.0 Hz), 7.25 (s, 1H, Ar-H),
7.22 (d, 2H, Ar-H, J = 7.2 Hz), 4.22 (s, 1H, C≡CH), 3.48 (s, 2H, Ar-CH2-N), 2.46 (br,
4H, 2×N-CH2), 0.98 (t, 6H, 2×CH3, J = 6.8 Hz). 13C-NMR (DMSO-d6): δ 157.6,
153.1, 153.0, 146.3, 140.3, 138.7, 138.3, 133.1, 129.7 (2×CH), 129.3, 128.9, 127.1,
127.0, 125.4, 123.3, 122.2, 118.7 (2×CH), 116.1, 110.4, 84.0, 81.0, 56.7, 46.4
(2×CH2), 11.9 (2×CH3). ESI-HRMS m/z: calc’d for C28H29N6O [M+H]+: 465.2403;
found: 465.2357.
4.1.37.
(R)-1-(4-((4-Methylpiperazin-1-yl)methyl)phenyl)-3-(4-((1-phenylethyl)
amino)quinazolin-6-yl)urea (2p)
Yield 25.6%. mp: 130.2-133.8 C. 1H-NMR (DMSO-d6): δ 8.94 (s, 1H, N-H),
8.92 (s, 1H, N-H), 8.39 (d, 1H, N-H, J = 8.4 Hz), 8.37 (s, 1H, Ar-H), 8.32 (s, 1H,
Ar-H), 7.80 (d, 1H, Ar-H, J = 8.4 Hz), 7.63 (d, 1H, Ar-H, J = 8.8 Hz), 7.46 (d, 4H,
Ar-H, J = 7.6 Hz), 7.31 (t, 2H, Ar-H, J = 7.2 Hz), 7.15-7.25 (m, 3H, Ar-H), 5.61 (t,
1H, Ar-CH-N, J = 6.8 Hz), 3.57 (s, 2H, Ar-CH2-N), 2.38 (br, 8H, 4×N-CH2), 2.21 (s,
13
3H, N-CH3), 1.60 (d, 3H, CH3, J = 7.2 Hz). C-NMR (DMSO-d6): δ 158.6, 153.8,
153.2, 145.7, 145.3, 138.9, 137.4, 131.9, 129.9 (2×CH), 128.6 (2×CH), 128.4, 127.0,
126.7 (2×CH), 126.5, 118.6 (2×CH), 115.6, 110.0, 62.0, 54.9 (2×CH2), 52.6, 49.7
(2×CH2), 45.8, 22.6. ESI-HRMS m/z: calc’d for C29H34N7O [M+H]+: 496.2825; found:
496.2819.
23

4.1.38.
(3a)
1-Phenyl-3-(4-((3-(trifluoromethyl)phenyl)amino)quinazolin-6-yl)urea
1
Yield 65.7% mp: 237.0-238.2 C. H-NMR (DMSO-d6): δ 10.02 (s, 1H, N-H),
9.02 (s, 1H, N-H), 8.92 (s, 1H, N-H), 8.58 (s, 1H, Ar-H), 8.56 (d, 1H, Ar-H, J = 2.0
Hz), 8.30 (s, 1H, Ar-H), 8.23 (d, 1H, Ar-H, J = 8.0 Hz), 7.89 (dd, 1H, Ar-H, J1 = 2.0
Hz, J2 = 8.8 Hz), 7.80 (d, 1H, Ar-H, J = 9.2 Hz), 7.63 (t, 1H, Ar-H, J = 8.0 Hz), 7.53
(d, 2H, Ar-H, J = 8.0 Hz), 7.46 (d, 1H, Ar-H, J = 8.0 Hz), 7.32 (t, 2H, Ar-H, J = 8.0
Hz), 7.01 (t, 1H, Ar-H, J = 7.6 Hz). 13C-NMR (DMSO-d6): δ 157.5, 153.1, 152.9,
146.3, 140.9, 140.0, 138.3, 130.0, 129.8, 129.5, 129.3 (2×CH), 129.0, 127.1, 126.1,
124.7 (d, JC-F = 270 Hz), 119.9, 118.8 (2×CH), 118.5, 116.1, 110.4. ESI-HRMS m/z:
calc’d for C22H17F3N5O [M+H] +: 424.1385; found: 424.1380.
4.1.39. 1-(4-Methylphenyl)-3-(4-((3-(trifluoromethyl)phenyl)amino)quinazolin-
6-yl)urea (3b)
1
Yield 68.1%. mp: 239.5-241.0 C. H-NMR (DMSO-d6): δ 10.02 (s, 1H, N-H),
8.99 (s, 1H, N-H), 8.82 (s, 1H, N-H), 8.57 (s, 1H, Ar-H), 8.55 (d, 1H, Ar-H, J = 2.0
Hz), 8.30 (s, 1H, Ar-H), 8.22 (d, 1H, Ar-H, J = 8.0 Hz), 7.88 (dd, 1H, Ar-H, J1 = 2.0
Hz, J2 = 8.8 Hz), 7.80 (d, 1H, Ar-H, J = 8.8 Hz), 7.63 (t, 1H, Ar-H, J = 8.0 Hz), 7.45
(d, 1H, Ar-H, J = 8.0 Hz), 7.41 (d, 2H, Ar-H, J = 8.4 Hz), 7.12 (d, 2H, Ar-H, J = 8.4
13
Hz), 2.26 (s, 3H, CH3). C-NMR (DMSO-d6): δ 157.5, 153.1, 152.8, 146.2, 140.9,
138.4, 137.4, 131.4, 130.0, 129.8, 129.7 (2×CH), 128.9, 127.1, 126.1, 124.7 (d, JC-F
=
270 Hz), 119.9, 118.9 (2×CH), 118.5, 116.1, 110.3, 20.8. ESI-HRMS m/z: calc’d for
C23H19F3N5O [M+H]+: 438.1542; found: 438.1536.
4.1.40.
1-(4-methoxyphenyl)-3-(4-((3-(trifluoromethyl)phenyl)amino)
quinazolin-6-yl)urea (3c)
1
Yield 61.7%. mp: 242.0-243.2 C. H-NMR (DMSO-d6): δ 10.02 (s, 1H, N-H),
8.97 (s, 1H, N-H), 8.76 (s, 1H, N-H), 8.57 (s, 1H, Ar-H), 8.54 (s, 1H, Ar-H), 8.30 (s,
1H, Ar-H), 8.22 (d, 1H, Ar-H, J = 8.0 Hz), 7.89 (d, 1H, Ar-H, J = 8.0 Hz), 7.79 (d, 1H,
Ar-H, J = 8.8 Hz), 7.63 (t, 1H, Ar-H, J = 8.0 Hz), 7.42-7.46 (m, 3H, Ar-H), 6.90 (d,
13
2H, Ar-H, J = 8.8 Hz), 3.73 (s, 3H, OCH3). C-NMR (DMSO-d6): δ 157.5, 155.1,
153.3, 152.7, 146.1, 140.9, 138.5, 133.0, 130.0, 129.8, 128.8, 127.1, 126.1, 124.7 (d,
24

JC-F = 266 Hz), 120.7 (2×CH), 120.0, 118.5, 116.1, 114.5 (2×CH), 110.2, 55.6.
ESI-HRMS m/z: calc’d for C23H19F3N5O2 [M+H]+: 454.1491; found: 454.1485.
4.1.41. General procedures for the synthesis of compounds 8a, 8b
The solution of 3-nitrobenzaldehyde (1.0 g, 6.62 mmol) and piperidine (or
pyrrolidine) (7.95 mmol) in dichloromethane (30 mL) was stirred at room temperature
for 2 h, then added portionwise triacetoxyborohydride (4.21 g, 19.86 mmol). The
mixture was stirred for another 3 h, added water (20 mL). The organic phase was
separated. The water phase was extracted with dichloromethane (20 mL  2). The
organic layer was combined, washed with brine (20 mL), dried (Na2SO4), filtered and
concentrated to produce compounds 8a, 8b as yellow oil.
4.1.42. 3-(Piperidin-1-ylmethyl)nitrobenzene (8a)
Yield 98.9%. 1H-NMR (CDCl3): δ 8.20 (s, 1H, Ar-H), 8.10 (d, 1H, Ar-H, J = 8.0
Hz), 7.69 (d, 1H, Ar-H, J = 8.0 Hz), 7.48 (t, 1H, Ar-H, J = 8.0 Hz), 3.55 (s, 2H,
Ar-CH2-N), 2.39 (br, 4H, 2×N-CH2), 1.56-1.62 (m, 4H, 2×CH2), 1.42-1.48 (m, 2H,
CH2).
4.1.43. 3-( Pyrrolidin-1-ylmethyl)nitrobenzene (8b)
Yield 98.0%. 1H-NMR (CDCl3): δ 8.20 (s, 1H, Ar-H), 8.10 (d, 1H, Ar-H, J = 8.0
Hz), 7.69 (d, 1H, Ar-H, J = 8.0 Hz), 7.48 (t, 1H, Ar-H, J = 8.0 Hz), 3.71 (s, 2H,
Ar-CH2-N), 2.51-2.54 (m, 4H, 2×N-CH2), 1.79-1.82 (m, 4H, 2×CH2).
4.1.44. General procedures for the synthesis of compounds 9a, 9b
Compounds 9a, 9b were prepared according to the reported method.33
4.1.45. 3-(Piperidin-1-ylmethyl)aniline (9a)
1
Yield 93.0%. mp: 96.7-97.8 C. H-NMR (CDCl3): δ 7.11 (t, 1H, Ar-H, J = 7.6
Hz), 6.73 (s, 1H, Ar-H), 6.72 (d, 1H, Ar-H, J = 8.0 Hz), 6.60 (d, 1H, Ar-H, J = 7.2 Hz),
3.66 (s, 2H, Ar-CH2-N), 2.41 (br, 4H, 2×N-CH2), 1.58-1.64 (m, 4H, 2×CH2),
1.42-1.48 (m, 2H, CH2).
4.1.46. 3-(Pyrrolidin -1-ylmethyl)aniline (9b)
1
Yield 93.2%. mp: 61.8-63.0 C. H-NMR (CDCl3): δ 7.11 (t, 1H, Ar-H, J = 7.6
Hz), 6.74 (s, 1H, Ar-H), 6.73 (d, 1H, Ar-H, J = 7.2 Hz), 6.60 (d, 1H, Ar-H, J = 8.0 Hz),
3.56 (s, 2H, Ar-CH2-N), 2.53-2.56 (m, 4H, 2×N-CH2), 1.80-1.83 (m, 4H, 2×CH2).
25

4.1.47. General procedures for the synthesis of compounds 2q and 2r
To a solution of 4-(3-trifluoromethylphenylamino)quinazolin-6-amine (0.12 g,
0.39 mmol) in acetonitrile (5 mL), CDI (0.24 g, 1.51 mmol) was added. The reaction
mixture was stirred at room temperature for 8 h. To this suspension, compound 9 in 5
mL of acetonitrile was added and stirred at room temperature for 2 h. The mixture
was then refluxed for another 2 h and cooled to room temperature. The off-white
precipitate was filtered and purified by chromatography on silica gel
(dichloromethane/methanol = 20:1, v/v) to give compound 2q or 2r as off-white
powder.
4.1.48.
1-(3-((Piperidin-1-yl)methyl)phenyl)-3-(4-((3-(trifluoromethyl)phenyl)
amino)quinazolin-6-yl)urea (2q)
1
Yield 31.3%. mp: 207.1-209.0 C. H-NMR (DMSO-d6): δ 10.02 (s, 1H, N-H),
8.97 (s, 1H, N-H), 8.93 (s, 1H, N-H), 8.58 (s, 1H, Ar-H), 8.56 (d, 1H, Ar-H, J = 2.4
Hz), 8.30 (s, 1H, Ar-H), 8.22 (d, 1H, Ar-H, J = 8.0 Hz), 7.89 (dd, 1H, Ar-H, J1 = 2.0
Hz, J2 = 8.8 Hz), 7.79 (d, 1H, Ar-H, J = 9.2 Hz), 7.63 (t, 1H, Ar-H, J = 8.0 Hz), 7.50
(s, 1H, Ar-H), 7.46 (d, 1H, Ar-H, J = 8.0 Hz), 7.38 (d, 1H, Ar-H, J = 8.0 Hz), 7.24 (t,
1H, Ar-H, J = 8.0 Hz), 6.93 (d, 1H, Ar-H, J = 8.0 Hz), 3.40 (s, 2H, Ar-CH2-N), 2.33
(br, 4H, 2×N-CH2), 1.51 (br, 4H, 2×CH2), 1.40 (br, 2H, CH2). 13C-NMR (DMSO-d6):
δ 157.5, 153.1, 152.9, 146.3, 140.9, 139.9, 138.3, 130.0, 129.8, 129.5, 129.1, 128.9,
127.1, 126.1, 124.7 (d, JC-F = 271 Hz), 123.1, 119.9, 119.1, 118.5, 117.4, 116.2, 110.3,
63.4, 54.4 (2×CH2), 25.9 (2×CH2), 24.4. ESI-HRMS m/z: calc’d for C28H28F3N6O
[M+H]+: 521.2277; found: 521.2271.
4.1.49.
1-(3-((Pyrrolidin-1-yl)methyl)phenyl)-3-(4-((3-(trifluoromethyl)phenyl)
amino)quinazolin-6-yl)urea (2r)
1
Yield 34.0%. mp: 204.0-206.5 C. H-NMR (DMSO-d6): δ 10.02 (s, 1H, N-H),
9.01 (s, 1H, N-H), 8.95 (s, 1H, N-H), 8.57 (s, 1H, Ar-H), 8.56 (d, 1H, Ar-H, J = 2.0
Hz), 8.30 (s, 1H, Ar-H), 8.22 (d, 1H, Ar-H, J = 8.0 Hz), 7.89 (dd, 1H, Ar-H, J1 = 2.0
Hz, J2 = 8.8 Hz), 7.79 (d, 1H, Ar-H, J = 9.2 Hz), 7.63 (t, 1H, Ar-H, J = 8.0 Hz), 7.56
(s, 1H, Ar-H), 7.46 (d, 1H, Ar-H, J = 8.0 Hz), 7.37 (d, 1H, Ar-H, J = 8.0 Hz), 7.25 (t,
1H, Ar-H, J = 8.0 Hz), 6.95 (d, 1H, Ar-H, J = 7.6 Hz), 3.59 (s, 2H, Ar-CH2-N), 2.49
26

(br, 4H, 2×N-CH2), 1.72 (br, 4H, 2×CH2). 13C-NMR (DMSO-d6): δ 157.5, 153.1,
152.9, 146.3, 140.9, 140.0, 138.3, 130.0, 129.8, 129.5, 129.1, 129.0, 127.1, 126.1,
124.7 (d, JC-F = 271 Hz), 122.8, 119.9, 118.9, 118.6, 117.4, 116.1, 110.3, 60.0, 53.9
(2×CH2), 23.5 (2×CH2). ESI-HRMS m/z: calc’d for C27H26F3N6O [M+H]+: 507.2120;
found: 507.2115.
4.1.50. General procedures for the synthesis of compounds 10a, 10b
Compounds 10a and 10b were prepared according to the reported process.42
4.1.51. 4-(4-Methylpiperazin-1-yl)nitrobenzene (10a)
1
Yield 92.7%. mp: 95.2-95.6 C. H-NMR (CDCl3): δ 8.15 (d, 2H, Ar-H, J = 9.2
Hz), 6.85 (d, 2H, Ar-H, J = 9.2 Hz), 3.48 (t, 4H, 2×N-CH2, J = 7.2 Hz), 1.25 (t, 4H,
2×N-CH2, J = 7.2 Hz), 2.40 (s, 3H, N-CH3).
4.1.52. 4-(Diethylamino)nitrobenzene (10b)
1
Yield 94.2%. mp: 64.0-64.8 C. H-NMR (CDCl3): δ 8.13 (d, 2H, Ar-H, J = 9.2
Hz), 6.61 (d, 2H, Ar-H, J = 9.2 Hz), 3.48 (q, 4H, 2×N-CH2, J = 4.8 Hz), 2.60 (t, 6H,
2×CH3, J = 4.8 Hz).
4.1.53. General procedures for the synthesis of compounds 11a and 11b
Compounds 11a and 11b were prepared according to the reported method.43
4.1.54. 4-(4-Methylpiperazin-1-yl)aniline (11a)
1
Yield 89.7%. mp: 72.0-73.4 C. H-NMR (CDCl3): δ 6.84 (d, 2H, Ar-H, J = 8.8
Hz), 6.78 (d, 2H, Ar-H, J = 8.8 Hz), 3.11 (t, 4H, 2×N-CH2, J = 4.8 Hz), 2.63 (t, 4H,
2×N-CH2, J = 4.8 Hz), 2.39 (s, 3H, N-CH3).
4.1.55. 4-(Diethylamino)aniline (11b)
1
Yield 85.2%. mp: 55.3-57.0 C. H-NMR (CDCl3): δ 7.80 (d, 2H, Ar-H, J = 9.2
Hz), 6.74 (d, 2H, Ar-H, J = 9.2 Hz), 3.45 (q, 4H, 2×N-CH2, J = 7.2 Hz), 1.10 (t, 6H,
2×CH3, J = 7.2 Hz).
4.1.56. General procedures for the synthesis of compounds 3d, 3e
Compounds 3d and 3e were synthesized by the same process with compound 2q.
4.1.57.
1-(4-(Diethylamino)phenyl)-3-(4-((3-(trifluoromethyl)phenyl)amino)
quinazolin-6-yl)urea (3d)
1
Yield 33.4%. mp: 182.0-184.0 C. H-NMR (DMSO-d6): δ 10.00 (s, 1H, N-H),
27

8.86 (s, 1H, N-H), 8.56 (s, 1H, N-H), 8.51 (s, 1H, Ar-H), 8.49 (s, 1H, Ar-H), 8.30 (s,
1H, Ar-H), 8.22 (d, 1H, Ar-H, J = 7.6 Hz), 7.89 (d, 1H, Ar-H, J = 8.4 Hz), 7.78 (d, 1H,
Ar-H, J = 8.8 Hz), 7.63 (t, 1H, Ar-H, J = 8.0 Hz), 7.45 (d, 1H, Ar-H, J = 7.2 Hz),
7.29 (d, 2H, Ar-H, J = 8.0 Hz), 6.65 (d, 2H, Ar-H, J = 8.0 Hz), 3.29 (q, 4H, 2×N-CH2,
J = 6.8 Hz), 1.07 (t, 6H, 2×CH3, J = 6.4 Hz). 13C-NMR (DMSO-d6): δ 157.5, 153.4,
152.7, 146.1, 144.1, 140.9, 138.8, 130.0, 129.8, 128.9, 128.5, 127.0, 126.0, 124.7 (d,
JC-F = 271 Hz), 121.5 (2×CH), 119.8, 118.5, 116.2, 112.9 (2×CH), 109.9, 44.3
(2×CH2), 12.8 (2×CH3). ESI-HRMS m/z: calc’d for C26H26F3N6O [M+H]+: 495.2120;
found: 495.2115.
4.1.58.
1-(4-(4-Methylpiperazin-1-yl)phenyl)-3-(4-((3-(trifluoromethyl)phenyl)
amino)quinazolin-6-yl)urea (3e)
1
Yield 37.5%. mp: 215.2-216.4 C. H-NMR (DMSO-d6): δ 10.00 (s, 1H, N-H),
8.91 (s, 1H, N-H), 8.65 (s, 1H, N-H), 8.56 (s, 1H, Ar-H), 8.53 (d, 1H, Ar-H, J = 2.0
Hz), 8.30 (s, 1H, Ar-H), 8.22 (d, 1H, Ar-H, J = 7.6 Hz), 7.88 (dd, 1H, Ar-H, J1 = 2.0
Hz, J2 = 8.8 Hz), 7.78 (d, 1H, Ar-H, J = 8.8 Hz), 7.63 (t, 1H, Ar-H, J = 8.0 Hz), 7.45
(d, 1H, Ar-H, J = 7.6 Hz), 7.36 (d, 2H, Ar-H, J = 8.8 Hz), 6.91 (d, 2H, Ar-H, J = 8.8
Hz), 3.07 (t, 4H, 2×N-CH2, J = 4.8 Hz), 2.46 (t, 4H, 2×N-CH2, J = 4.8 Hz), 2.23 (s,
3H, N-CH3). 13C-NMR (DMSO-d6): δ 157.5, 153.2, 152.8, 147.1, 146.2, 140.9, 138.6,
131.9, 130.0, 129.5, 129.0, 127.0, 126.1, 124.7 (d, JC-F = 271 Hz), 120.3 (2×CH),
119.9, 118.5, 116.6 (2×CH), 116.2, 110.1, 55.1 (2×CH2), 49.2 (2×CH2), 46.1.
ESI-HRMS m/z: calc’d for C27H27F3N7O [M+H]+: 522.2229; found: 522.2224.
4.1.59. General procedures for the synthesis of compounds 4a and 4b
To a solution of 6-amino-4-anilinoquinazoline or 4-(1-phenylethyl)quinazoline-
4,6-diamine (0.23 mmol) in acetonitrile (5 mL), CDI (0.11 g, 0.69 mmol) was added.
The reaction mixture was stirred at room temperature for 8 h when lots of solid
produced. To this suspension, the solution of 2-(4-methylpiperazin-1-yl)ethylamine in
5 mL of acetonitrile was added. Then, the mixture was stirred at room temperature for
2 h, refluxed for another 2 h, cooled to room temperature and concentrated under
reduced pressure. The residue was added water (20 mL) and the mixture was
extracted with dichloromethane (20 mL  3). The organic layer was combined,
28

washed with brine (20 mL), dried (Na2SO4), concentrated under reduced pressure and
purified by chromatography on silica gel (dichloromethane/methanol = 15:1, v/v) to
give compounds 4a or 4b as yellow powder.
4.1.60.
1-(2-(4-Methylpiperazin-1-yl)ethyl)-3-(4-((3-(trifluoromethyl)phenyl)
amino)quinazolin-6-yl)urea (4a)
1
Yield 45.5%. mp: 169.8-170.5 C. H-NMR (DMSO-d6): δ 9.96 (s, 1H, N-H),
9.03 (s, 1H, N-H), 8.54 (s, 1H, N-H), 8.46 (d, 1H, Ar-H, J = 2.0 Hz), 8.29 (s, 1H,
Ar-H), 8.21 (d, 1H, Ar-H, J = 8.0 Hz), 7.83 (dd, 1H, Ar-H, J1 = 2.0 Hz, J2 = 8.8 Hz),
7.74 (d, 1H, Ar-H, J = 8.8 Hz), 7.62 (t, 1H, Ar-H, J = 8.0 Hz), 7.44 (d, 1H, Ar-H, J =
7.6 Hz), 6.35 (t, 1H, Ar-H, J = 5.2 Hz), 3.20-3.29 (m, 4H, 2×CH2), 2.41-2.49 (m, 8H,
4×N-CH2), 2.27 (s, 3H, N-CH3). 13C-NMR (DMSO-d6): δ 157.4, 155.6, 152.5, 145.9,
141.0, 139.2, 130.0, 129.5, 128.8, 126.7, 124.7 (d, JC-F = 261 Hz), 126.0, 119.8, 118.4,
116.2, 109.3, 57.6, 54.7 (2×CH2), 52.4 (2×CH2), 45.5, 36.9. ESI-HRMS m/z: calc’d
for C23H27F3N7O [M+H]+: 474.2229; found: 474.2224.
4.1.61.
(R)-1-(2-(4-Methylpiperazin-1-yl)ethyl)-3-(4-((1-phenylethyl)amino)
quinazolin-6-yl)urea (4b)
1
Yield 33.6%. mp: 101.3-103.5 C. H-NMR (DMSO-d6): δ 8.89 (s, 1H, N-H),
8.30 (s, 1H, N-H), 8.28 (s, 1H, N-H), 8.26 (d, 1H, Ar-H, J = 2.0 Hz), 7.74 (dd, 1H,
Ar-H, J1 = 2.0 Hz, J2 = 8.8 Hz), 7.58 (d, 1H, Ar-H, J = 8.8 Hz), 7.45 (d, 2H, Ar-H, J
= 7.2 Hz), 7.31 (t, 2H, Ar-H, J = 8.0 Hz), 7.21 (t, 1H, Ar-H, J = 7.2 Hz), 6.29 (t, 1H,
Ar-H, J = 4.2 Hz), 5.50-5.64 (m, 1H, Ar-CH-N), 3.21-3.30 (m, 4H, 2×CH2), 2.36-2.46
13
(m, 8H, 4×N-CH2), 2.23 (s, 3H, N-CH3), 1.59 (d, 3H, CH3, J = 7.2 Hz). C-NMR
(DMSO-d6): δ 158.5, 155.7, 153.4, 145.4, 145.2, 138.3, 128.6 (2×CH), 128.3, 127.0,
126.7 (2×CH), 126.1, 115.7, 109.9, 57.7, 54.9 (2×CH2), 52.7 (2×CH2), 49.7, 45.8,
36.9, 22.6. ESI-HRMS m/z: calc’d for C24H32N7O [M+H]+: 434.2668; found:
434.2663.
4.2. Biological materials and methods
4.2.1. Antiproliferative assay
The in vitro antiproliferative activities of compounds were determined by MTT
(3-[4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium bromide) assay. A431 or
29