Discovery of 1-(3-aryl-4-chlorophenyl)-3-(p-aryl)urea derivatives against breast cancer by inhibiting PI3K/Akt/mTOR and Hedgehog signalings

Article abstract of DOI:10.1016/j.ejmech.2017.09.002

PI3K/Akt/mTOR and hedgehog (Hh) signalings are two important pathways in breast cancer, which are usually connected with the drug resistance and cancer migration. Many studies indicated that PI3K/Akt/mTOR inhibitors and Hh inhibitors displayed synergistic effects, and the combination of the two signaling drugs could delay drug resistance and inhibit cancer migration in breast cancer. Therefore, the development of molecules simultaneously inhibiting these two pathways is urgent needed. Based on the structures of PI3K inhibitor buparlisib and Hh inhibitor vismodegib, a series of hybrid structures were designed and synthesized utilizing rational drug design and computer-based drug design. Several compounds displayed excellent antiproliferative activities against several breast cancer cell lines, including triple-negative breast cancer (TNBC) MDA-MB-231 cell. Further mechanistic studies demonstrated that the representative compound 9i could inhibit both PI3K/Akt/mTOR and hedgehog (Hh) signalings by inhibiting the phosphorylation of S6K and Akt as well as decreasing the SAG elevated expression of Gli1. Compound 9i could also induce apoptosis remarkably in T47D and MDA-MB-231 cells. In the transwell assay, 9i showed significant inhibition on the migration of MDA-MB-231.

Full text of DOI:10.1016/j.ejmech.2017.09.002

Accepted Manuscript
Discovery of 1-(3-aryl-4-chlorophenyl)-3-(p-aryl)urea derivatives against breast cancer
by inhibiting PI3K/Akt/mTOR and Hedgehog signalings
Wenlu Li, Qinsheng Sun, Lu Song, Chunmei Gao, Feng Liu, Yuzong Chen, Yuyang
Jiang
PII:
S0223-5234(17)30684-0
10.1016/j.ejmech.2017.09.002
EJMECH 9717
DOI:
Reference:
To appear in: European Journal of Medicinal Chemistry
Received Date: 15 April 2017
Revised Date: 31 August 2017
Accepted Date: 2 September 2017
Please cite this article as: W. Li, Q. Sun, L. Song, C. Gao, F. Liu, Y. Chen, Y. Jiang, Discovery of
1-(3-aryl-4-chlorophenyl)-3-(p-aryl)urea derivatives against breast cancer by inhibiting PI3K/Akt/
mTOR and Hedgehog signalings, European Journal of Medicinal Chemistry (2017), doi: 10.1016/
j.ejmech.2017.09.002.
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ACCEPTED MANUSCRIPT
Discovery of 1-(3-aryl-4-chlorophenyl)-3-(p-aryl)urea derivatives against breast
cancer by simultaneously inhibiting PI3K/Akt/mTOR and Hedgehog signalings

ACCEPTED MANUSCRIPT
Discovery of 1-(3-aryl-4-chlorophenyl)-3-(p-aryl)urea derivatives against breast
cancer by inhibiting PI3K/Akt/mTOR and Hedgehog signalings
a,b
c,b
a,b
b,d
b,d*
Wenlu Li , Qinsheng Sun , Lu Song , Chunmei Gao , Feng Liu , Yuzong
d,e
b,c,d*
Chen , Yuyang Jiang
a
Department of Chemistry, Tsinghua University, Beijing 100084
The Ministry-Province Jointly Constructed Base for State Key Lab-Shenzhen Key
Laboratory of Chemical Biology, the Graduate School at Shenzhen, Tsinghua
University, Shenzhen 518055
b
c
Department of Pharmacology and Pharmaceutical Sciences, School of Medicine,
Tsinghua University, Beijing, 100084, P. R. China
d
National & Local United Engineering Lab for Personalized anti-tumor drugs, the
Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055
e
Bioinformatics and Drug Design Group, Department of Pharmacy, Centre for
Computational Science and Engineering, National University of Singapore, 117543,
Singapore
*
Corresponding author. Tel.: +86 755 2603 2094; fax: +86 755 2603 2094;
Email address: liu.feng@sz.tsinghua.edu.cn; jiangyy@sz.tsinghua.edu.cn.

ACCEPTED MANUSCRIPT
Abstract:
PI3K/Akt/mTOR and hedgehog (Hh) signalings are two important pathways in
breast cancer, which are usually connected with the drug resistance and cancer
migration. Many studies indicated that PI3K/Akt/mTOR inhibitors and Hh inhibitors
displayed synergistic effects, and the combination of the two signaling drugs could
delay drug resistance and inhibit cancer migration in breast cancer. Therefore, the
development of molecules simultaneously inhibiting these two pathways is urgent
needed. Based on the structures of PI3K inhibitor buparlisib and Hh inhibitor
vismodegib, a series of hybrid structures were designed and synthesized utilizing
rational drug design and computer-based drug design. Several compounds displayed
excellent antiproliferative activities against several breast cancer cell lines, including
triple-negative breast cancer (TNBC) MDA-MB-231 cell. Further mechanistic studies
demonstrated that the representative compound 9i could inhibit both PI3K/Akt/mTOR
and hedgehog (Hh) signalings by inhibiting the phosphorylation of S6K and Akt as
well as decreasing the SAG elevated expression of Gli1. Compound 9i could also
induce apoptosis remarkably in T47D and MDA-MB-231 cells. In the transwell assay,
9
i showed significant inhibition on the migration of MDA-MB-231.
Keywords: Breast cancer; drug design; antitumor activity; PI3K; mTOR; hedgehog.
1
. Introduction
Breast cancer is the most common cause of cancer death in women. Though

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approximately 75% of breast cancers express oerstrogen receptor (ER) and can be
treated with ER inhibitors [1, 2], the recurring problems are acquired resistance and
cancer metastasis. Another challenge is that there are no drugs approved by the FDA
(US Food and Drug Administration) to treat the triple-negative breast cancer (TNBC).
Therefore, much more work needs to be done to provide more effective approaches
for breast cancer treatment. [3-4]
PI3K/Akt/mTOR pathway is essential in growth, metabolism, proliferation,
differentiation, motility, survival and angiogenesis [5, 6]. Aberrantly activation of
PI3K/Akt/mTOR is one of the most mutationally altered pathways in breast cancer. [7,
8
] Many compounds targeting PI3K/Akt/mTOR pathway have been approved or in
clinical/preclinical studies. [9-17] Buparlisib (Fig. 1), a PI3K inhibitor, is currently
entering phase III clinical trials for solid cancers [18]. PF-05212384 (Fig. 1), a dual
PI3K/mTOR inhibitor, has been demonstrated with manageable safety profile and
promising antitumor activity for advanced solid malignancies in Phase II. Rapamycin,
a selective mTOR inhibitor, has been approved as a chemotherapeutic agent by FDA.
[
19] Preclinical and clinical evidence indicated that PI3K/Akt/mTOR inhibitors could
increase the benefit of endocrine therapy of ER+ breast cancer. [20] However, these
therapeutic approaches have encountered intrinsic and acquired resistance. [21] A
great crosstalk in tumor pathways is one of the main reasons of drug resistance.
Consequently, combination therapies have been applied in preclinical and clinical
studies to provide better cancer treatments. [5]
The hedgehog (Hh) pathway is critical for embryonic proliferation and

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differentiation, which is relatively dormant in adults, but aberrantly activated in
tumorigenesis. [22-26] Therefore, targeting the Hh pathway has been a promising
strategy for the treatment of cancer. [27, 28] Hh pathway consists of several important
proteins, among which the Smoothened (Smo) and Gli1 have attracted great attentions.
[
29-38] Now, a series of compounds targeting Smo and Gli1 have been developed,
some of which have been approved or entered into clinical status, such as vismodegib
27, 39, 40], sonidegib [41] (Fig. 1), etc. Recent studies indicated that inhibiting both
[
PI3K/Akt/mTOR and hedgehog signalings could achieve better treatments for breast
cancer, and the cooperation of PI3K/Akt/mTOR and Hh signaling inhibitors have
shown synergistic effect on inhibiting breast cancer cell proliferation and invasion.
[
42-46] Remarkably, the combination therapy using sonidegib and buparlisib treating
metastatic breast cancer has been in phase I study (NCT01576666). Inspired by the
results above, the development of molecules simultaneously inhibiting
PI3K/Akt/mTOR and Hh signalings may be an effective strategy for the treatment of
breast cancer, which may avoid the drug-drug interaction risks of drug combination.
However, up to now, only few researches have been reported. [47]
Fig 1. The structures of PI3K/mTOR inhibitors and Hh inhibitors.
Our group has been focusing on the development of multitargeted antitumor

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agents, and some multifunctional inhibitors with good anticancer activities have been
successfully synthesized. [48-54] Herein, we reported a series of rationally designed
derivatives as PI3K/Akt/mTOR and Hh multifunctional inhibitors, and the
bioactivities against tumor cells were also evaluated.
2
. Result and discussion
2
.1 Design strategy
Curis [55] obtained a series of dual Hh and HDAC inhibitors by introducing a
zinc binding motif to vismodegib analogs, which indicated that replacing the B ring of
vismodegib with other substituents, the inhibitory activity against Hh signaling could
be reserved. In addition, PF-05212384 had better activity against PI3K/Akt/mTOR
signaling than buparlisib, which indicated that the introduction of an appropriate
group to the amino group of buparlisib might retain or even improve the inhibition
activity against PI3K/Akt/mTOR signaling. Based on these results, we intended to
combine buparlisib and the A part of vismodegib by a urea linker to afford the
unsymmetrical diaryl urea compound 9a (Fig. 2). Since diaryl urea was quite a
common structure in both PI3K/mTOR and Smo inhibitors [37], compound 9a might
be a potential PI3K/Akt/mTOR and Hh signaling inhibitor.
Fig. 2 Design strategy of PI3K/mTOR and Hh multifunctional inhibitors.

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Molecular docking was firstly performed to evaluate whether the designed 9a
was a potent PI3K, mTOR and Smo inhibitor, respectively (Fig. 3). For Smo protein,
as expected, 9a could insert into the deep pocket of Smo. The nitrogen atoms of
pyrimidine could form hydrogen bonds with Phe 222, while the nitrogen atom of
pyridine formed a hydrogen bond with Lys 395. For PI3K protein, the nitrogen atom
of the morpholine in 9a could form two hydrogen bonds with Val882 and Lys 833 of
PI3K, while formed two hydrogen bonds with Asp 2195 and Val 2240 of mTOR. The
docking results revealed that 9a could fit pretty well with the target proteins, and thus
it could be a potent inhibitor for both PI3K/Akt/mTOR and Hh signalings. Therefore,
9
a and its derivatives 9b-9u were designed and synthesized.
Fig. 3 The best-scored docking poses of 9a within Smo protein (A, PDB code: 4O9R),
PI3Kα (B, PDB code: 1E7V) and mTOR (C, PDB code: 4J6T).
2
.2 Chemistry
The unsymmetrical diaryl ureas were synthesized according to the synthetic
routes as shown in Scheme 1. The commercially available 4-nitrochlorobenzene (1),
which was a weak nucleophile, could afford 1-chloro-2-iodo-4-nitro-benzene (2) after
treatment with NIS in the presence of trifluoromethanesulfonic acid [56]. The
reduction of compound 2 gave 4-chloro-3-iodoaniline (3). Compounds 5 could be

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afforded by Suzuki reactions in different conditions. As boronic acid substituted at 2-
and 3- positions of pyridines were not available, iodoarenes 3 should be transformed
to boric acid ester and then reacted with 2-brominepyridine or 3-brominepyridine
under the condition of NaHCO and Pd (PPh ) in reflux 1,4-dioxane/H O to give 5a
3
3 4
2
or 5b. The transformation of compound 3 to 4 was achieved with
,4,4',4',5,5,5',5'-Octamethyl-2,2-bi(1,3,2-dioxaborolane), KOAc and PdCl (dppf) in
4
2
reflux DMF. Compound 5d, 5e and 5f could be afforded by Suzuki reactions of
iodoarenes 3 with the corresponding boric acid similar to the reaction condition of 5a.
Compounds 7 were afforded by one step of necleophilic reactions of morphline with
Et N in CH Cl . The reactions of 6a and 6c with morphline could also be achieved in
3
2
2
EtOH, while 6d would react with EtOH and gave byproduct
4
,4'-(6-ethoxy-1,3,5-triazine-2,4-diyl)dimorpholine. Treatment of compounds 7 with
-aminophenylboronicacidpinacolester afforded compounds 8 according to the
4
preparation condition of 5a. To furnish target compounds 9, compounds 5 should be
firstly transformed to isocyanates with triphosgene and DIEA in anhydrous CH Cl
2
2
and then reacted with another amines 8. During the process of preparing isocyanates,
three points should be paid attention to avoid the symmetrical diaryl ureas byproducts.
First, the reaction temperature should keep at -78 °C all along. Second, the amines 5
dissolved in CH Cl with DIEA should be added dropwise into the solution of
2
2
triphosgene in CH Cl . Third, the process should avoid water. According to the
2
2
methods described above, twenty-one compounds were synthesized and identified by
1
infrared spectra (IR), high-resolution mass spectra (HRMS), elemental analysis, H

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1
3
NMR, C NMR, COSY and HMBC spectra.
Scheme 1. Reagents and conditions: (a) NIS, CF SO H, rt, 24 h; (b) Fe, NH Cl, H O,
3
3
4
2
EtOH, 75 °C, 3 h; (c) arylboronic acid, Pd(Ph P) , NaOH or Na CO ,
3
4
2
3
1
4
9
2
,4-dioxane/H O,
110
°C,
24h;
(d)
2
,4,4',4',5,5,5',5'-Octamethyl-2,2-bi(1,3,2-dioxaborolane), KOAc, PdCl (dppf), DMF,
2
5 °C, 2 h; (e) 2- or 3-bromopyridine, Pd (PPh ) , NaHCO 1,4-dioxane/H O, 110 °C,
3
4
3,
2
4 h; (f) morpholine, Et N, EtOH, rt, 24 h; (g) 4-aminophenylboronicacidpinacolester,
3
Pd (Ph P) , NaHCO , 1,4-dioxane/H O, 110 °C, 24 h； (h) morpholine, Et N, CH Cl ,
3
4
3
2
3
2
2
rt, 4 h; (i) 4-aminophenylboronicacidpinacolester, Pd (Ph P) , NaHCO ,
3
4
3
1
,4-dioxane/H O, 110 °C, overnight; (j) Bis(trichloromethyl) carbonate, DIEA,
2
CH Cl , -78 °C, 2 h; (k) 2-chloro-4-methylsulphonylbenzoic acid, HATU, DIEA,
2
2
DMF, rt, overnight.
2
.3 Inhibition of cell proliferation
All the newly synthesized 1-(3-aryl-4-chlorophenyl)-3-(p-aryl)urea derivatives
were screened for their inhibitory effects against T47D by MTT assay, and the IC50

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values were summarized in Table 1. All the compounds showed modest or significant
cytotoxicity in T47D cell lines. For compounds 9a-9r, 3-pyridyl or 4-pyridyl as the R1
group (9b, 9c, 9h, 9i, 9n) had better cell antiproliferative activity than phenyl or
thienyl derivatives, some of which displayed similar to or even better antiprolifertive
activity than buparlisib (IC = 6.92 µM). For example, 3-pyridyl substituted 9b (IC
5
0
50
=
3.96 µM) had the best activity among 9a-9f, which was about 9-folds of 2-pyridyl
substituted 9a (IC = 35.11 µM), 6-folds of 2-thienyl substituted 9d (IC = 25.44
5
0
50
µM), and 4-folds of phenyl substituted 9d (IC = 15.57 µM) and 3-thienyl substituted
5
0
9
g (IC = 18.69 µM). So the pyridyl group was important for the loss of cell viability.
50
For compounds 9s-9u, their activities (with the IC50 values about 20 µM) were
relatively lower than the corresponding compounds bearing A part of vismodegib
(with the IC50 value about 10 µM), which indicated that keeping the B part of
vismodegib was not preferred. Switching the pyrimidine core (9a) to triazine (9g), the
IC values of 2-pyridyl substituted compounds were decreased about 60% (9a vs 9g),
5
0
while those of phenyl substituted compounds were increased 76% (9e vs 9k).
Eliminating one of the two morpholine groups did not change the IC value a lot for
5
0
most compounds, such as 9b vs 9n, except for 2-pyridyl derivatives (9a vs 9m) and
benzyl derivatives (9d vs 9p). Especially, compound 9b (IC = 3.96 µM) and 9n
5
0
(
IC50 = 4.11 µM) showed better antiprolifertive activity than the positive control
drugs.
Table 1. Cytotoxicity of the compounds 9a-9u against T47D cells.

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R
N
2
R
2
X
N
X
N
Cl
O
N
Cl
O
N
N
O
N
O
O
S
H
R
1
N
N
H
H
O
9s-9u
9a-9r
a
Data are expressed as the mean ± SD from the dose response curves of at least three
independent experiments.
Further cytotoxicity assays against MCF-7, MDA-MB-231 and PC-3 cells, were
tested only with the outstanding candidates in T47D cytotoxicity assays, including
2
-pyridyl (9b, 9h, 9n) or 3-pyridyl (9c, 9i, 9o) substituted compounds (Table 2). All
of them showed significant antiproliferative activity against these cancer cells. Those
compounds with two morpholine groups (9b-9c and 9h-9i) showed greater
cytotoxicity (below 10 µM) than those with only one morpholine group (9n-9o). For

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MCF-7 cells, four compounds had better antiproliferative activities than buparlisib
(
IC = 11.05 µM), especially 9h with the IC value of 5.37 µM. For the TNBC cells,
50 50
MDA-MB-231, the cytotoxicity of our compounds was still reserved. The best
candidate inhibitor was 9i, with the IC value of 3.61 µM. The cytotoxicity against
5
0
another classic PI3K/Akt/mTOR signaling activated cell line (PC-3) was also detected.
The compound 9h showed satisfied antiproliferative activity with IC value of 5.37
5
0
µM. As a whole, our compounds could effectively inhibit the proliferation of selected
cell lines, including TNBC MDA-MB-231 cells.
Table 2. Cytotoxicity of the selected compounds against MCF-7, MDA-MB-231 and
PC-3 cells.
a
Data are expressed as the mean ± SD from the dose response curves of at least three
independent experiments.
2
.4 Inhibition of PI3K/Akt/mTOR and Hh signalings
Compounds with good antiproliferative activity against cancer cells were
selected to further evaluate their activities on disrupting the PI3K/Akt/mTOR and Hh
signalings. First, the kinase assays were performed to evaluate the inhibition abilities

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of our compounds against PI3K and mTOR (Table 3). All of the selected compounds
showed relatively low PI3K inhibitory rate at the concentration of 5 µM. But the IC50
values against mTOR were in nanomolecular range. Comparing the results of 9g-9j,
the inhibition efficiency sequence was 2-pyridyl > 4-pyridyl > 3-pyridyl > benzyl (9g >
9
i > 9h > 9n). 2-Pyridyl substituted 9g had the lowest IC50 of 90 nM. 3-Pyridyl
groups (9b, 9h) and 4-pyridyl groups (9c, 9i) were tolerated for the mTOR activities,
while phenyl group (9j) resulted in ten times decreased activity. Reducing two
morpholine groups to one, the mTOR activity was lost (9n). So the pyridine and the
two-morpholine groups were all necessary for the inhibition of mTOR.
Table 3. Enzymatic activities of compounds against mTOR and PI3Kα.
a
The inhibition rate at 50 µM was 30.00%.
To further determine whether our compounds would suppress the activation of
both PI3K/Akt/mTOR and Hh signalings in cancer cell, western blot assay was
performed to test the expression of relative proteins. The phosphorylation levels of
Akt and S6K were signatures of PI3K/Akt/mTOR pathway, so the most potent

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compound 9i for MDA-MB-231, was selected to test for its suppressive effects on
p-S6K (Thr 412 and Thr 389) and p-Akt (Ser 473) in MDA-MB-231 cells. For Hh
pathway, both the Smo agonist SAG and the representative compound 9i were used to
treat the cells to see whether 9i could inhibit the elevated expression level of Gli1
resulted by SAG. The suppressive effects on S6K and Akt phosphorylation were
conducted at the concentration of 5 µM for 1 h, 2 h and 4 h. As illustrated in Fig. 4A
and 4B, the inhibitory effect of 9i on S6K phosphorylation was almost complete in 4
h and the expression of p-Akt was also decreased in a time-dependent manner. The
main effect of 9i on PI3K/Akt/mTOR signaling was inhibiting the phosphorylation of
S6K at Thr 389. As shown in Fig. 4C and 4D, the MDA-MB-231 cells treated with 5
µM SAG had higher Gli1 expression, which could be inhibited by 9i. The inhibition
extent of SAG elevated Gli1 expression deepened with the increasing concentration of
9
i. The results further demonstrated 9i could down-regulate both PI3K/Akt/mTOR
and hedgehog signalings.
Fig. 4. Compound 9i inhibited the relative protein expressions of PI3K/Akt/mTOR

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and Hh signalings. (A) The inhibition effects of compound 9i (5 µM) on the
expression of p-S6K (Thr 412), p-S6K (Thr 389) and p-Akt (Ser 473) in
MDA-MB-231 cells are depicted. Equal loading was testified by the detection of
β-actin. (B) The percentages of p-S6K (Thr 412), p-S6K (Thr 389) and p-Akt (Ser
4
73) expressions in cells treated with 9i for different time. (C) The inhibition effects
of compound 9i on the expression of SAG (5 µM) elevated Gli1 in MDA-MB-231
cells. (D) The percentages of Gli1 expressions in cells treated with 9i.
2
.5 Inducing apoptosis and inhibiting cell migration
Both of the PI3K/Akt/mTOR and Hh signalings have been reported to have
effect on the apoptosis pathway by targeting related protein. [57-59] To evaluate
whether compound 9i could induce apoptosis, an Annexin-V/PI binding assay was
conducted in MDA-MB-231 and T47D cells as shown in Fig.5. For T47D cells, the
percentage of early stage of apoptosis cells was increased from 4.49% to 75.1% when
treated with 5 µM of 9i. For MDA-MB-231 cells, as the concentration of 9i increased,
the percentage of both early stage and late stage of apoptotic cells increased. Almost
9
5% of the cells were in the process of apoptosis when the drug concentration was 10
µM. In conclusion, compound 9i had greater effect on MDA-MB-231 cells than T47D
cells and the mechanism of apoptosis induction may be different.

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Fig. 5 Compound 9i induced apoptosis. Analysis was carried out on a flow cytometer
according to the presented protocols. Fluorescence of the FITC/PI-stained T47D cells
(A) and MDA-MB-231 cells (B) was measured using CyAnTMADP Analyzer
(Beckman Coulter, Inc.) and Summit software.
Both PI3K/Akt/mTOR and Hh signalings have been reported to connect with
breast cancer invasion and metastasis as well. [60, 61] In this study, we used transwell
migration assay to evaluate the ability of compound 9i inhibiting cell migration. The
results, as shown in Fig 6., indicated that the cells travelling through the filter were
decreased with the concentration of 9i increased, which indicated that 9i could
effectively inhibit the invasion of tumor cells.

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Fig. 6. Compound 9i inhibited the penetration of MDA-MB-231 cells through the
filter membrane.
3
. Conclusion
Based on the structures of vismodegib and buparlisib, a novel series of
unsymmetrical diaryl ureas, as potent inhibitors simultaneously inhibiting
PI3K/Akt/mTOR and Hh signalings, were designed and synthesized. The synthesized
compounds were evaluated in three breast cancer cell lines, including T47D, MCF-7,
and MDA-MB-231. Several compounds displayed comparable or better
antiproliferative activities than buparlisib, among which 9b, 9h and 9i showed high
inhibitory activities against T47D, MCF-7 and MDA-MB-231 cells respectively.
Among all the compounds, 9i showed the best antiproliferative activity against
MDA-MB-231 cells and also had high cytotoxicity against T74D and MCF-7 cells.
The enzymatic activity of 9i against mTOR was in nanomolecular value. Additionally,
compound 9i could inhibit the expression of p-S6K, p-Akt and SAG elevated Gli1,
induce tumor apoptosis and also inhibit migration of TNBC MDA-MB-231. As a
whole, all the results indicated that 9i could be a lead compound for further
development.

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4
4
4
. Experimental protocols
.1 Chemistry and chemical methods
.1.1 1-Chloro-2-iodo-4-nitro-benzene (2)
4
-Chloronitrobenzene (1, 10 mmol) was dissolved in CF SO H (50 mmol) with
3 3
ice bath, after which NIS (10 mmol) was added in small portion. The reaction mixture
was stirred at room temperature for 24 h and quenched with ice water. Extracted the
aqueous solution with CH Cl for three times. Combined the organic extracts and
2
2
washed them with 10% sodium bisulfite solution and water. The organic layer was
1
condensed to give a brown solid, yield 99%. H NMR (400 MHz, Chloroform-d) δ
13
8
.71 (d, J = 2.6 Hz, 1H), 8.17 (dd, J = 8.8, 2.6 Hz, 1H), 7.62 (d, J = 8.8 Hz, 1H). C
NMR (101 MHz, CDCl ) δ 146.26, 145.86, 135.13, 129.62, 124.20, 98.03.
3
4
.1.2 4-Chloro-3-iodoaniline (3)
A mixture of compound 2 (10 mmol), iron powder (30 mmol) and saturated
NH Cl aqueous (10 mL) in EtOH (40 mL) was stirred at 75 °C for 3 h. The reaction
4
mixture was filtered through celite and the residue was washed with CH Cl . The
2
2
filtrate was evaporated to remove most of EtOH, and then dissolved in CH Cl . The
2
2
organic mixture was washed with water and brine, dried over Na SO , and condensed
2
4
1
to give a yellow solid, yield 97%. H NMR (400 MHz, DMSO-d ) δ 7.14 (dd, J = 5.6,
6
1
3
3
.0 Hz, 2H), 6.58 (dd, J = 8.6, 2.7 Hz, 1H), 5.42 (s, 2H). C NMR (101 MHz, DMSO)
δ 149.28, 129.58, 124.68, 123.10, 115.86, 98.96.
.1.3 4-Chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (4)
4

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A mixture of compound 3 (5 mmol), 4,4,4',4',5,5,5',5'-Octamethyl-2,2-
bi(1,3,2-dioxaborolane) (10 mmol), KOAc (10 mmol), PdCl (dppf) (0.1 mmol) in
2
DMF (20mL) was stirred at 95 °C for 2 h under Ar. The reaction mixture was filtered
through celite. The filtrate was concentrated in vacuo and then extracted with CH Cl .
2
2
The organic layer was evaporated to give a residue, which was purified by
1
chromatography (petroleum ether/CH Cl , 10:1) to afford a white solid, yield 50%. H
2
2
NMR (400 MHz, DMSO-d ) δ 7.10 (d, J = 8.5 Hz, 1H), 6.97 (d, J = 2.9 Hz, 1H), 6.70
6
1
3
(
dd, J = 8.5, 2.9 Hz, 1H), 5.32 (s, 2H), 1.38 (s, 12H). C NMR (101 MHz, DMSO) δ
1
47.40, 129.97, 124.63, 121.85, 118.00, 84.08 (2C), 25.08 (4CH3).
.1.4 4-Chloro-3-(pyridin-2-yl)aniline (5a)
4
A mixture of compound 4 (5 mmol), 2-bromopyridine (10 mmol), Pd(Ph P) (0.1
3
4
mmol), NaHCO (15 mmol) in 1,4-dioxane/H O (20mL) was stirred at 110 ℃ for 24
3
,
2
h under Ar. The reaction mixture was filtered through celite. The filtrate was
concentrated in vacuo and then extracted with EtOAc. The organic layer was
evaporated to give a residue, which was purified by chromatography (petroleum
1
ether/EtOAc, 5:1) to give pure product as a white solid, yield 64%. H NMR (400
MHz, DMSO-d ) δ 8.64 (m, J = 4.9, 1.8, 1.0 Hz, 1H), 7.85 (td, J = 7.7, 1.8 Hz, 1H),
6
7
.57 (dt, J = 7.9, 1.1 Hz, 1H), 7.37 (m, J = 7.6, 4.8, 1.2 Hz, 1H), 7.14 (d, J = 8.6 Hz,
13
1
H), 6.76 (d, J = 2.8 Hz, 1H), 6.61 (m, J = 8.6, 2.8 Hz, 1H), 5.36 (s, 2H). C NMR
(101 MHz, DMSO) δ 157.40, 149.67, 148.31, 139.57, 136.57, 130.53, 124.84, 122.97,
1
17.26, 117.02, 115.72.
4
.1.5 4-Chloro-3-(pyridin-3-yl)aniline (5b)

ACCEPTED MANUSCRIPT
Similar procedure of 5a was performed to give compound 5b as a white solid,
1
yield 51%. H NMR (400 MHz, DMSO-d ) δ 8.64 – 8.47 (m, 2H), 7.81 (dt, J = 7.8,
6
1
.9 Hz, 1H), 7.46 (m, J = 7.9, 4.8, 0.9 Hz, 1H), 7.18 (d, J = 8.4 Hz, 1H), 6.72 – 6.47
1
3
(
m, 2H), 5.41 (s, 2H). C NMR (101 MHz, DMSO) δ 149.74, 148.95, 148.65, 137.07,
1
37.03, 135.76, 130.63, 123.67, 117.54, 116.68, 115.54.
4
.1.6 4-Chloro-3-(pyridin-4-yl)aniline (5c)
A mixture of compound 3 (5 mmol), 4-pyridylboronic acid (10 mmol), Pd(Ph P)
3
4
(
0.1 mmol), NaOH (10 mmol) in 1,4-dioxane/H O (20mL) was stirred at 110 ℃
2
overnight under Ar. The reaction mixture was filtered through celite. The filtrate was
concentrated in vacuo and then extracted with CH Cl . The organic layer was
2
2
evaporated to give a residue, which was purified by chromatography (CH Cl /CH OH,
2
2
3
1
3
0:1) to give pure product as a white solid, yield 53%. H NMR (400 MHz, DMSO-d )
6
δ 8.72 – 8.53 (m, 2H), 7.48 – 7.34 (m, 2H), 7.19 (d, J = 8.5 Hz, 1H), 6.72 – 6.53 (m,
1
3
2
1
4
H), 5.44 (s, 2H). C NMR (101 MHz, DMSO) δ 150.22 (2CH), 148.95, 147.83,
38.15, 131.02, 124.73 (2CH), 117.13, 116.45, 116.10.
.1.7 6-Chloro-[1,1'-biphenyl]-3-amine (5d)
A mixture of compound 3 (5 mmol), phenylboronic acid (10 mmol), Pd(Ph P)4
3
(
0.1 mmol), Na CO (10 mmol) in 1,4-dioxane/H O (20mL) was stirred at 110 ℃ for
2 3 2
2
4h under Ar. The reaction mixture was filtered through celite. The filtrate was
concentrated in vacuo and then extracted with EtOAc. The organic layer was
evaporated to give a residue, which was purified by chromatography (petroleum
1
ether/EtOAc, 10:1) to give pure product as a yellow liquid, yield 88%. H NMR (400

ACCEPTED MANUSCRIPT
MHz, Chloroform-d) δ 7.48 – 7.30 (m, 5H), 7.19 (s, 1H), 6.62 (d, J = 2.7 Hz, 1H),
1
3
6
1
1
4
.56 (dd, J = 8.5, 2.9 Hz, 1H), 3.65 (s, 2H). C NMR (101 MHz, CDCl ) δ 145.24,
3
41.11, 139.75, 130.55, 129.41 (2CH), 128.06 (2CH), 127.58, 121.57, 117.84, 115.37,
15.36.
.1.8 4-Chloro-3-(thiophen-2-yl)aniline (5e)
Similar procedure of 5d was performed to give compound 5e as a yellow liquid,
1
yield 75%. H NMR (400 MHz, DMSO-d ) δ 7.60 (dd, J = 5.1, 1.3 Hz, 1H), 7.34 (dd,
6
J = 3.7, 1.2 Hz, 1H), 7.26 – 7.05 (m, 2H), 6.82 (d, J = 2.8 Hz, 1H), 6.58 (dd, J = 8.6,
1
3
2
1
4
.7 Hz, 1H), 5.40 (s, 2H). C NMR (101 MHz, DMSO) δ 148.52, 140.68, 132.78,
31.16, 127.82, 127.75, 126.97, 117.26, 116.35, 115.32.
.1.9 4-Chloro-3-(thiophen-3-yl)aniline (5f)
Similar procedure of 5d was performed to give compound 5e as a yellow liquid,
1
yield 75%. H NMR (400 MHz, DMSO-d ) δ 7.58 (d, J = 4.8 Hz, 2H), 7.27 (dd, J =
6
4
2
1
4
.6, 1.7 Hz, 1H), 7.13 (d, J = 8.5 Hz, 1H), 6.69 (d, J = 2.7 Hz, 1H), 6.56 (dd, J = 8.5,
1
3
.8 Hz, 1H), 5.31 (s, 2H). C NMR (101 MHz, DMSO) δ 148.38, 140.12, 135.22,
30.68, 129.15, 126.01, 124.43, 117.61, 116.55, 114.90.
.1.10 4,4'-(6-Chloropyrimidine-2,4-diyl)dimorpholine (7a)
To the solution of compound 6a (10 mmol) and Et N (24 mmol) in EtOH (50
3
mL), morpholine (20 mmol) was added dropwise at 0 ℃ . The mixture was stirred at
room temperature for 24 h and a white solid was precipitated. Concentrated the
reaction mixture to remove EtOH. The residue was dissolved in CH Cl , washed with
2
2
water and brine. The organic layer was concentrated to give the residue, which was

ACCEPTED MANUSCRIPT
purified using flash chromatography (petroleum ether/EtOAc, 7:1) to yield a white
1
solid, yield 78%. H NMR (400 MHz, Chloroform-d) δ 5.87 (s, 1H), 3.96 – 3.18 (m,
1
3
1
6
4
6H). C NMR (101 MHz, CDCl ) δ 163.54, 160.88, 160.60, 91.19, 66.84 (2CH ),
3
2
6.51 (2CH ), 44.44 (2CH ), 44.37 (2CH ).
2
2
2
.1.11 4,4'-(6-Chloropyrimidine-1,3,5-triazine-2,4-diyl)dimorpholine (7b)
Similar procedure of 7a was performed to give compound 7b as a white solid,
1
13
yield 58%. H NMR (400 MHz, DMSO-d ) δ 3.58 (dt, J = 31.5, 4.5 Hz, 16H). C
6
NMR (101 MHz, DMSO) δ 170.65, 166.06, 66.42 (4CH ), 43.85 (4CH ).
2
2
4
.1.12 4-(4-Chloropyrimidin-2-yl)morpholine (7c)
To the solution of compound 6c (10 mmol) and Et N (12 mmol) in CH Cl (50
3
2
2
mL), morpholine (10 mmol) was added dropwise at 0 ℃ . The mixture was stirred at
room temperature for 4 h and then concentrated. The residue was dissolved in CH Cl ,
2
2
washed with water and brine. The organic layer was concentrated to give the residue,
1
which was purified using chromatography to yield a white solid, yield 53%. H NMR
(400 MHz, Chloroform-d) δ 8.08 (d, J = 6.1 Hz, 1H), 6.40 (d, J = 6.2 Hz, 1H), 3.87 –
1
3
3
6
4
.54 (m, 8H). C NMR (101 MHz, CDCl ) δ 162.88, 160.74, 157.36, 101.16, 101.14,
3
6.35 (2CH ), 44.31 (2CH ).
2
2
.1.13 4-(2,6-Dimorpholinopyrimidin-4-yl)aniline (8a)
A mixture of compound 7a (5 mmol), 4-aminophenylboronic acid pinacol ester
(
10 mmol), Pd(Ph P) (0.1 mmol), Na CO (10 mmol) in 1,4-dioxane/H O (20 mL)
3 4 2 3 2
was stirred at 110 ℃ for 24h under Ar. The reaction mixture was filtered through
celite. The filtrate was concentrated in vacuo and then extracted with EtOAc. The

ACCEPTED MANUSCRIPT
organic layer was evaporated to give a residue, which was purified by
chromatography (petroleum ether/EtOAc, 5:1) to give pure product as a yellow solid,
1
yield 59%. H NMR (400 MHz, DMSO-d ) δ 7.84 (d, J = 8.3 Hz, 2H), 6.58 (d, J = 8.2
6
1
3
Hz, 2H), 6.47 (s, 1H), 5.48 (s, 2H), 3.74 – 3.51 (m, 16H). C NMR (101 MHz,
DMSO) δ 164.08, 163.54, 161.58, 151.19, 128.38 (2CH), 125.59, 113.64 (2CH),
8
7.14, 66.70 (4CH ), 44.61 (4CH ).
2 2
4
.1.14 4-(4,6-Dimorpholino-1,3,5-triazin-2-yl)aniline (8b)
Similar procedure of 8a was performed to give compound 8b as a white solid,
1
yield 52%. H NMR (400 MHz, DMSO-d ) δ 8.05 (d, J = 8.6 Hz, 2H), 6.57 (d, J = 8.6
6
13
Hz, 2H), 5.70 (s, 2H), 3.93 – 3.52 (m, 16H). C NMR (101 MHz, DMSO) δ 169.95,
1
4
4
65.07, 152.77, 130.25 (2CH), 126.46, 123.90, 114.80, 113.22 (2CH), 66.56 (4CH2),
3.74 (4CH2).
.1.15 4-(2-Morpholinopyrimidin-4-yl)aniline (8c)
Similar procedure of 8a was performed to give compound 8c as a white solid,
1
yield 53%. H NMR (400 MHz, DMSO-d ) δ 8.21 (d, J = 6.1 Hz, 1H), 8.11 – 7.96 (m,
6
1
3
2
H), 6.70 – 6.51 (m, 3H), 5.52 (s, 2H), 3.80 – 3.52 (m, 8H). C NMR (101 MHz,
DMSO) δ 163.46, 162.10, 156.52, 151.51, 129.55 (2CH), 125.83, 113.56 (2CH),
1
00.45, 66.39 (2CH ), 44.20 (2CH ).
2 2
4
.1.16 1-(4-Chloro-3-(pyridin-2-yl)phenyl)-3-(4-(2,6-dimorpholinopyrimidin-4-yl)-
phenyl)urea (9a)
The mixture of compound 5a (1 mmol) and DIEA(1 mmol) in anhydrous CH Cl
2
2
(5mL) was added dropwise to the bis(trichloromethyl) carbonate (0.5 mmol) in

ACCEPTED MANUSCRIPT
anhydrous CH Cl (5 mL) at -78 ℃ . After 1 h, the temperature was allowed to warm
2
2
up to room temperature. The mixture of compound 7a (1 mmol) and DIEA (1 mmol)
in anhydrous CH Cl (5mL) was added in one portion. The mixture was stirred at
2
2
room temperature for another 2 h and then quenched with saturated Na CO . Extract
2
3
the aqueous solution with CH Cl for three times. The organic layer was concentrated
2
2
to give the residue, which was purified using flash chromatography (petroleum
ether/THF, 1:1) to yield a white solid, yield 30%. HRMS (ESI) m/z calculated for
+
[
M+H] 572.2177, found 572.2183, 574.2207; mp 208-209 ℃ ; IR (KBr): ν 3300,
max
-1 1
2
969, 2855, 1645, 1573, 1539, 1309, 1241, 1189, 1116 cm . H NMR (400 MHz,
DMSO-d ) δ 9.05 (s, 1H), 9.03 (s, 1H), 8.76 (d, J = 4.9 Hz, 1H), 8.13 (s, 1H), 8.13 –
6
8
8
3
1
1
.11 (d, J = 7.8, 1H), 7.83 (d, J = 2.3 Hz, 1H), 7.72 (d, J = 7.8 Hz, 1H), 7.59 (d, J =
.6 Hz, 2H), 7.54 (d, J = 3.2 Hz, 2H), 7.49 (dd, J = 7.6, 4.9 Hz, 1H), 6.69 (s, 1H),
1
3
.83 – 3.62 (m, 16H). C NMR (101 MHz, DMSO) δ 164.10, 162.69, 161.64, 156.62,
52.78 (C=O), 149.92, 141.62, 139.59, 139.23, 136.89, 132.09, 130.68, 127.90 (2CH),
25.00, 124.04, 123.39, 121.49, 120.19, 118.23 (2CH), 88.53, 66.67 (2CH ), 66.46
2
(
2CH ), 44.60 (2CH ), 44.58 (2CH ). Analysis calculated (%): C, 62.99; H, 5.29; N,
2 2 2
1
4
1
7,14; O, 8.39; found (%): C, 62.79; H, 5.29; N, 16.83; O, 8.98.
.1.17
-(4-Chloro-3-(pyridin-3-yl)phenyl)-3-(4-(2,6-dimorpholinopyrimidin-4-yl)phenyl)ure
a (9b)
Similar procedure of 9a was performed to give compound 9c as a white solid,
+
yield 23%. HRMS (ESI) m/z calculated for [M+H] 572.2177, found 572.2187,

ACCEPTED MANUSCRIPT
o
5
1
8
1
1
1
1
74.2222; mp 200-201 C; IR (KBr): νmax 3302, 2970, 2855, 1643, 1573, 1539, 1309,
-
1 1
241, 1189, 1116 cm . H NMR (400 MHz, DMSO-d ) δ 9.04 (s, 1H), 9.00 (s, 1H),
6
.65 (s, 1H), 8.63 (d, J = 2.8 Hz, 1H), 8.08 (d, J = 1.9 Hz, 1H), 8.06 (d, J = 2.1 Hz,
H), 7.90 (m, J = 7.9, 2.0 Hz, 1H), 7.63 (d, J = 1.6 Hz, 1H), 7.54 (m, 1H), 7.52 (d, J =
1
3
.8 Hz, 4H), 6.64 (s, 1H), 3.82 – 3.49 (m, 16H). C NMR (101 MHz, DMSO) δ
64.12, 162.71, 161.66, 152.79 (C=O), 149.83, 149.36, 141.60, 139.55, 137.21,
37.17, 135.13, 132.15, 130.70, 127.90 (2CH), 124.43, 123.82, 121.30, 120.07,
1
18.23 (2CH), 88.55, 66.67 (2CH ), 66.46 (2CH ), 44.62 (2CH ), 44.59 (2CH ).
2 2 2 2
Analysis calculated (%): C, 62.99; H, 5.29; N, 17,14; O, 8.39; found (%): C, 62.58; H,
5
4
1
.26; N, 16.73; O, 9.01.
.1.18
-(4-Chloro-3-(pyridin-4-yl)phenyl)-3-(4-(2,6-dimorpholinopyrimidin-4-yl)phenyl)ure
a (9c)
Similar procedure of 9a was performed to give compound 9c as a white solid,
+
yield 24%. HRMS (ESI) m/z calculated for [M+H] 572.2177, found 572.2178,
o
5
1
8
6
1
1
74.2210; mp 221-223 C; IR (KBr): νmax 3328, 2965, 2858, 1649, 1597, 1539, 1308,
-
1 1
258, 1189, 1116 cm . H NMR (400 MHz, DMSO-d6) δ 9.05 (s, 1H), 9.01 (s, 1H),
.69 (d, J = 5.8 Hz, 2H), 8.07 (d, J = 8.7 Hz, 2H), 7.65 (s, 1H), 7.63 – 7.41 (m, 6H),
1
3
.64 (s, 1H), 3.78 – 3.51 (m, 16H). C NMR (101 MHz, DMSO-d ) δ 164.10, 161.63,
6
52.78 (C=O), 150.15 (2CH), 146.87, 141.59, 137.96, 132.13, 130.84, 127.91 (2CH),
24.55 (2CH), 123.77, 120.83, 120.36, 118.23 (2CH), 88.56, 66.67 (2CH ), 65.41
2

ACCEPTED MANUSCRIPT
(
2CH ), 44.61 (2CH ), 44.59 (2CH ). Analysis calculated (%): C, 62.99; H, 5.29; N,
2
2
2
1
4
1
7,14; O, 8.39; found (%): C, 62.78; H, 5.32; N, 17.12; O, 8.73.
.1.19
-(6-Chloro-[1,1'-biphenyl]-3-yl)-3-(4-(2,6-dimorpholinopyrimidin-4-yl)phenyl)urea
(9d)
Similar procedure of 9a was performed to give compound 9d as a white solid,
+
yield 35%. HRMS (ESI) m/z calculated for [M+H] 572.2227, found 571.2227,
o
5
1
8
=
73.2260; mp 207-208 C; IR (KBr): νmax 3369, 2959, 2852, 1641, 1573, 1537, 1307,
-
1 1
243, 1187, 1116 cm . H NMR (400 MHz, DMSO-d ) δ 9.01 (s, 1H), 8.96 (s, 1H),
6
.07 (d, J = 8.3 Hz, 1H), 8.04 (d, J = 8.3 Hz, 1H), 7.59 (d, J = 2.1 Hz, 1H), 7.53 (d, J
1
3
8.4 Hz, 2H), 7.47 – 7.42 (m, 7H), 3.66 (td, J = 18.7, 5.0 Hz, 16H). C NMR (101
MHz, DMSO) δ 164.12, 162.76, 161.68, 152.83 (C=O), 141.66, 140.53, 139.43,
1
1
4
39.33, 132.12, 130.54, 129.56 (2CH), 128.73 (2CH), 128.26, 127.88 (2CH), 124.25,
21.34, 119.41, 118.25 (2CH), 88.54, 66.67 (2CH ), 66.46 (2CH ), 44.63 (2CH ),
2
2
2
4.60 (2CH ). Analysis calculated (%): C, 65.20; H, 5.47; N, 14.72; O, 8.40; found
2
(%): C, 65.12; H, 5.56; N, 14.32; O, 8.93.
4
1
.1.20
-(4-Chloro-3-(thiophen-2-yl)phenyl)-3-(4-(2,6-dimorpholinopyrimidin-4-yl)phenyl)u
rea (9e)
Similar procedure of 9a was performed to give compound 9e as a white solid,
+
yield 32%. HRMS (ESI) m/z calculated for [M+H] 577.1789, found 577.1805,
o
5
79.1813; mp 199-201 C; IR (KBr): νmax 3361, 2959, 2852, 1644, 1573, 1537, 1307,

ACCEPTED MANUSCRIPT
-
1 1
1
7
8
243, 1187, 1116 cm . H NMR (400 MHz, DMSO-d ) δ 8.95 (s, 2H), 7.99 (s, 1H),
6
.97 (d, J = 8.3 Hz, 1H), 7.71 (d, J = 2.5 Hz, 1H), 7.64 – 7.56 (m, 1H), 7.45 (dd, J =
.9, 3.1 Hz, 2H), 7.39 (dd, J = 8.9, 3.1 Hz, 2H), 7.33 (dd, J = 3.6, 1.4 Hz, 1H), 7.10
1
3
(
dd, J = 5.2, 3.5 Hz, 1H), 6.55 (s, 1H), 3.58 (m, 16H). C NMR (101 MHz, DMSO) δ
1
1
8
64.12, 162.74, 161.67, 152.81 (C=O), 141.62, 139.81, 139.49, 132.99, 132.15,
31.21, 128.38, 128.01, 127.91 (2CH), 127.74, 123.88, 120.92, 119.56, 118.24 (2CH),
8.56, 66.67 (2CH ), 66.47 (2CH ), 44.63 (2CH ), 44.60 (2CH ). Analysis calculated
2
2
2
2
(%): C, 60.36; H, 5.07; N, 14.56; O, 8.32; S, 5.56; found (%): C, 60.22; H, 5.41; N,
1
4
1
5.29; O, 8.62; S, 5.63.
.1.21
-(4-Chloro-3-(thiophen-3-yl)phenyl)-3-(4-(2,6-dimorpholinopyrimidin-4-yl)phenyl)u
rea (9f)
Similar procedure of 9a was performed to give compound 9f as a white solid,
+
yield 40%. HRMS (ESI) m/z calculated for [M+H] 577.1789, found 577.1784,
o
5
1
8
79.1796; mp 227-228 C; IR (KBr): νmax 3361, 2959, 2852, 1644, 1573, 1539, 1308,
-1
1
241, 1187, 1115 cm . H NMR (400 MHz, DMSO-d ) δ 9.01 (s, 1H), 8.94 (s, 1H),
6
.07 (s, 1H), 8.06 (d, J = 8.3 Hz, 1H), 7.71 (dd, J = 3.0, 1.3 Hz, 1H), 7.66 (d, J = 5.1
Hz, 1H), 7.66 (s, 1H), 7.54 (d, J = 8.3 Hz, 2H), 7.52 (d, J = 8.3 Hz, 2H), 7.34 (dd, J =
1
3
5
1
1
6
.0, 1.4 Hz, 1H), 6.63 (s, 1H), 3.67 (m, 16H). C NMR (101 MHz, DMSO) δ 164.13,
62.77, 161.68, 152.82 (C=O), 141.67, 139.36, 139.31, 135.38, 132.12, 130.72,
29.06, 127.89 (2CH), 126.45, 125.16, 124.26, 121.13, 119.31, 118.24 (2CH), 88.55,
6.67 (2CH ), 66.46 (2CH ), 44.64 (2CH ), 44.60 (2CH ). Analysis calculated (%): C,
2
2
2
2

ACCEPTED MANUSCRIPT
6
8
4
1
0.36; H, 5.07; N, 14.56; O, 8.32; S, 5.56; found (%): C, 61.71; H, 5.22; N, 14.76; O,
.62; S, 5.53.
.1.22
-(4-Chloro-3-(pyridin-2-yl)phenyl)-3-(4-(4,6-dimorpholino-1,3,5-triazin-2-yl)phenyl
)
urea (9g)
Similar procedure of 9a was performed to give compound 9g as a white solid,
+
yield 28%. HRMS (ESI) m/z calculated for [M+H] 573.2129, found 573.2142,
o
5
1
8
75.2199; mp 260 C (decomposed); IR (KBr): νmax 3336, 2965, 2858, 1649, 1597,
-1 1
527, 1307, 1258, 1173, 1111 cm . H NMR (400 MHz, DMSO-d ) δ 9.06 (s, 1H),
6
.99 (s, 1H), 8.70 (d, J = 4.9 Hz, 1H), 8.27 (d, J = 8.5 Hz, 2H), 7.92 (td, J = 7.8, 1.9
Hz, 1H), 7.78 (d, J = 2.3 Hz, 1H), 7.67 (d, J = 7.9 Hz, 1H), 7.55 (d, J = 8.5 Hz, 2H),
1
3
7
.53 – 7.46 (m, 2H), 7.44 (dd, J = 7.6, 4.8 Hz, 1H), 3.99 – 3.46 (m, 16H). C NMR
(101 MHz, DMSO) δ 169.56, 165.20 (2C), 156.67, 152.71 (C=O), 149.91, 143.20,
1
1
39.66, 139.18, 136.83, 130.68, 129.51 (2CH), 124.95, 124.19, 123.34, 121.61,
20.28, 117.94 (2CH), 66.54 (4CH ), 43.85 (4CH ). Analysis calculated (%): C, 60.78;
2
2
H, 5.10; N, 19.55; O, 8.38; found (%): C, 60.52; H, 5.28; N, 20.01; O, 8.46.
4
1
.1.23
-(4-Chloro-3-(pyridin-3-yl)phenyl)-3-(4-(4,6-dimorpholino-1,3,5-triazin-2-yl)phenyl
)
urea (9h)
Similar procedure of 9a was performed to give compound 9h as a white solid,
+
yield 19%. HRMS (ESI) m/z calculated for [M+H] 573.2129, found 573.2125,
o
5
75.2135; mp 232 C (decomposed); IR (KBr): νmax 3336, 2965, 2858, 1649, 1597,

ACCEPTED MANUSCRIPT
-
1
1
1
8
1
1
1
4
527, 1307, 1258, 1173, 1111 cm . H NMR (400 MHz, DMSO-d6) δ 8.66 (ds, 2H),
.23 (d, J = 8.4 Hz, 2H), 7.64 (s, 1H), 7.58 (d, J = 8.8 Hz, 3H), 7.47 (d, J = 8.5 Hz,
1
3
H), 7.43 (d, J = 5.0 Hz, 2H), 3.85 – 3.63 (m, 16H). C NMR (101 MHz, DMSO) δ
69.55, 165.14 (2C), 153.03 (C=O), 150.11, 146.91, 143.56, 139.93, 137.89, 130.74,
30.43, 129.43 (2CH), 124.50, 123.56, 120.83, 120.34, 117.87 (2CH), 66.53 (4CH2),
3.81 (4CH ). Analysis calculated (%): C, 60.78; H, 5.10; N, 19.55; O, 8.38; found
2
(%): C, 60.97; H, 5.34; N, 19.95; O, 8.39.
4
1
.1.24
-(4-Chloro-3-(pyridin-4-yl)phenyl)-3-(4-(4,6-dimorpholino-1,3,5-triazin-2-yl)phenyl
)
urea (9i)
Similar procedure of 9a was performed to give compound 9i as a white solid,
+
yield 21%. HRMS (ESI) m/z calculated for [M+H] 573.2129, found 573.2134,
o
5
1
9
1
1
75.2115; mp 210 C (decomposed); IR (KBr): νmax 3336, 2965, 2858, 1649, 1597,
-1 1
527, 1307, 1258, 1173, 1111 cm . H NMR (400 MHz, DMSO-d6) δ 9.21 (s, 1H),
.12 (s, 1H), 8.64 (t, J = 4.5 Hz, 2H), 8.27 (d, J = 8.4 Hz, 2H), 7.89 (d, J = 7.9 Hz,
H), 7.63 (s, 1H), 7.56 (d, J = 8.5 Hz, 2H), 7.52 (d, J = 2.9 Hz, 3H), 3.92 – 3.60 (m,
1
3
6H). C NMR (101 MHz, DMSO) δ 169.52, 165.14 (2C), 152.76 (C=O), 149.81
2CH), 143.20, 139.52, 137.20, 137.15, 135.13, 130.68, 129.50 (2CH), 124.51,
23.81, 121.37 (2CH), 120.15, 117.93 (2CH), 66.53 (4CH ), 43.81 (4CH ). Analysis
(
1
2
2
calculated (%): C, 60.78; H, 5.10; N, 19.55; O, 8.38; found (%): C, 61.03; H, 5.28; N,
9.67; O, 8.45.
1