Design, synthesis and bioactivities of phenylamino-pyrimidine derivatives as novel protein tyrosine kinase inhibitors

Article abstract of DOI:10.2174/157018011796235202

A series of novel phenylamino-pyrimidine derivatives were designed and synthesized as antitumor agent based on the lead compound of Imatinib by application of the principle of bioisosterism, hybridization and structural optimization. The bioactivities of the new target compounds were tested against human KU812 cells in vitro, some target compounds show promising activities and can be considered for further development.

Full text of DOI:10.2174/157018011796235202

596
Letters in Drug Design & Discovery, 2011, 8, 596-601
Design, Synthesis and Bioactivities of Phenylamino-Pyrimidine Derivatives
as Novel Protein Tyrosine Kinase Inhibitors
Xinfu Hong1,2, Xianzhao Kuang1,2, Guohua Ding1,2, Yilin Liu1,2, Jinlin Liu1,2, Yanjin Zhao1 and
Shuxin Li*,1,2
1Institute of Radiation and Irradiation Medicine, Academy of Military Medical Science, Beijing 100850, China;
2Department of Medicinal Chemistry, Jiangxi University of Traditional Chinese Medicine, Nanchang 330006, China
Received January 02, 2011: Revised April 26, 2011: Accepted April 29, 2011
Abstract: A series of novel phenylamino-pyrimidine derivatives were designed and synthesized as antitumor agent
based on the lead compound of Imatinib by application of the principle of bioisosterism, hybridization and structural op-
timization. The bioactivities of the new target compounds were tested against human KU812 cells in vitro, some target
compounds show promising activities and can be considered for further development.
Keywords: Protein tyrosine kinase, Phenylamino-pyrimidine, Synthesis, Bioactivity.
INTRODUCTION
target compounds (5a-d, 6a-d) were not obtained (Scheme
1). To our astonishment, HNMR and LC/MS showed that
1
Protein kinases play an important role in signal transduc-
tion pathways, regulating a number of cellular functions such
as cell proliferation, differentiation, migration and cell death
[1-3]. So the inhibition of protein kinases, especially protein
tyrosine kinases (PTKs), have been recognized as attractive
cell-signaling targets for drug discovery in the treatment of
cancer and other diseases [4-7]. During the past few years,
progress has been made aiming at finding selective PTKs
inhibitors. A number of small molecule compounds, espe-
cially the class of 2-phenylaminopyrimidine compounds,
such as Imatinib (STI571) [8-12] and Nilotinib (AMN107)
[13-14], functioning at the molecular level with high speci-
ficity, shows promise as a therapeutic agent for the treatment
of chronic myelogenous leukemia (CML). But as STI571 is
widely used, drug-resistance seriously hampers therapeutic
efficacy [15-16]. Hence, our study was to design and synthe-
size a series of novel phenylamino-pyrimidine derivatives
based on the lead compound of Imatinib by application of
the principle of bioisosterism, hybriddization and structural
optimization (Fig. 1).
chloro atom was introduced in the pyrimidine ring and the
new compounds (7a-g, 8a-g) were obtained. So in this paper,
besides the original target compounds (5a-d, 6a-d), we also
synthesized the new compounds (7a-g, 8a-g) and tested their
bioactivities.
N
O
N
N
H
N
N
H
Imatinib
N
N
R
N
N
A
B
H
R1
N
N
N
H
C
target compounds
O
O
R2
water-soluble group
was introduced
CHEMISTRY
The strategy for the synthesis of the target compounds
began with the 4,6-dichloro-2-methyl-pyrimidine which was
aminated with intermediates (1a-b) to afford intermediates
(2a-b) in high yields. Amination of intermediates (2a-b)
with morpholine in 1,4-dioxane gave intermediates (3a-b),
which were hydrolyzed with LiOH in the solution of water
and methanol to afford intermediates (4a-b). Amidation of
the intermediates (4a-b) with substituented phenylamino
through CDI gave the target compounds (5a-d, 6a-d) [17];
but when we tried to synthesize the target compounds (5a-d,
6a-d) by amidation of the intermediates (4a-b) with substitu-
ented phenylamino through thionyl chloride, the original
Fig. (1). structure of the target compounds.
GENERAL
All reagents were purchased from commercial sources
and used without further purification. Melting points were
measured in open capillaries and are uncorrected. H-NMR
was recorded in DMSO-d6 on a Bruker Avance 400 spec-
trometer; chemical shifts (ꢀ) are reported in parts per million
(ppm) relative to tetramethylsilane (TMS), used as an inter-
nal standard. Mass spectra (MS) were obtained from Agilent
1100 LC/MS Spectrometry Services.
1
RESULTS
*Address correspondence to this author at the Institute of Radiation and
Irradiation Medicine, Academy of Military Medical Science Beijing
100850, China; Tel/Fax: +86-010-66932289; E-mail: lisx28@163.com
In our attempts to obtain compounds with more potent
bioactivities and lower drug-reisistance, a series of novel
1570-1808/11 $58.00+.00
© 2011 Bentham Science Publishers Ltd.

Design, Synthesis and Bioactivities of Phenylamino-Pyrimidine Derivatives
Letters in Drug Design & Discovery, 2011, Vol. 8, No. 7 597
R
R
N
N
N
N
a
b
+
Cl
N
H
COOCH3
H2N
Cl
COOCH3
Cl
2a-b
1a R=H
1b R=CH3
R
R
N
N
N
N
c
N
N
H
COOH
N
COOCH3
N
H
O
O
4a-b
3a-b
R
R=H
5a
R1=4-CH3
N
N
N
R2=H
H
d
R1
N
R=H
R=H
5b
5c
R1=4-F
R1=4-I
R2=H
N
H
R2
R2=H
O
O
5d R=H
R1=2-Cl
R1=4-CH3
R2=6-CH3
R2=H
5a-d, 6a-d
6a R=CH3
6b R=CH3
R1=4-F
R1=4-I
R2=H
R=CH3
6c
R2=H
6d R=CH3
R1=2-Cl
R2=6-CH3
7a R=H
R
N
R1=4-CH3
R1=4-F
R2=H
R2=H
N
N
H
R=H
R=H
7b
7c
R1
N
e
N
H
R2
R1=4-I
R2=H
Cl
O
O
7d R=H
7e R=H
R1=2-Cl
R2=6-CH3
R2=H
7a-g, 8a-g
R1=3-CF3
R1=4-OCH3
R1=4-OCF3
R=H
R=H
7f
R2=H
R2=H
7g
R=CH3
8a
R1=4-CH3
R2=H
8b R=CH3
8c R=CH3
R1=4-F
R2=H
R1=4-I
R2=H
R=CH3
8d
R1=2-Cl
R1=3-CF3
R1=4-OCH3
R2=6-CH3
R2=H
8e R=CH3
8f R=CH3
8g R=CH3
R2=H
R1=4-OCF3
R2=H
Scheme 1. Synthesis of the target compounds; Reagents and condition: a) con.HCl, isopropanol; b) morpholine, 1,4-dioxane; c) LiOH, H2O,
methanol, 2mol/LHCl; d) CDI, substituented phenylamino; e) thionyl chloride; substituented phenylamino.
phenylamino-pyrimidine derivatives were synthesized and
evaluated as potent PTKs inhibitors. The antitumor activity
of the new target compounds were determined against hu-
man KU812 cells in vitro, and the results were shown in Ta-
ble 1. Compared with STI-571, some of the target com-
pounds obviously showed excellent bioactivities, especially
the compounds 7b and 7e. Compounds 7d and 7g showed
equivalent bioactivities with STI-571. Analyzing the bioac-
tivities and their structures, the following structure-activity
relationship was gained. Simply through introduction of a
methyl in the pyrimidine ring(A ring), the inhibition against
PTKs was enhanced, but the methyl(when R=CH3) group in
B ring had little effect on their bioactivities. In these pheny-
lamino-pyrimidine derivatives, the introduction of chloro

598 Letters in Drug Design & Discovery, 2011, Vol. 8, No. 7
Hong et al.
Table 1. Bioactivities of the Target Compounds Against KU812 Cell In Vitro
Compound
IC50 (μM)
Compound
IC50 (μM)
STI-571
5a
0.86
17.61
1.65
-
-
6a
6b
6c
6d
8a
8b
8c
8d
8e
8f
>50.0
15.80
>100.0
11.42
>50.0
20.71
>100.0
2.69
5b
5c
>100.0
15 .60
5.12
5d
7a
7b
0.51
7c
30.86
0.90
7d
7e
0.41
10.31
>100.0
8.7
7f
>50.0
0.78
7g
8g
KU812 cells (purchased from American Type Culture Collection) cultured in a RPMI-1640 medium (manufactured by Sigma) containing 10% (v/v) fetal calf serum (FCS) (manufac-
tured by Sigma) (RPMI-1640/FCS). KU812 cells were seeded at density of 5000 cells/100 μl/well and 4000 cells/100μl/well in each of 96-hole-plate (manufactured by costar),
respectively. The plate was incubated in a CO2 incubator overnight. A test drug was prepared with dimethylsulfoxide (DMSO) (manufactured by Nacalai Tesque) in the concentra-
tion 1000 times higher than the test concentration (0, 0.00001 to 1μM). The resulting solution was diluted 500 times in a RPMI-1640/FCS medium and then 100 μl of the diluent was
added in a well. The plate was incubated in a CO2 incubator. After 72 hours, 20μl of Cell counting Kit-8 (5 mmol/l WST-8, 0.2 mmol/l 1-Methoxy PMS, 150 mmol/l NaCl) (manu-
factured by Dojindo) was added to each well. After reaction for color development in a CO2 incubator for 3 hours, an absorbance offermazan, generated by reduction of WST-8 was
determined at 450 nm using Multilevel counter ARVOsx (manufactured by Wallac). In the RPMI-1640/FCS medium containing 0.1% DMSO, when absorbance of a region in which
cells after culturing in the CO2 incubator for 72 hours were seeded is defined as a cell growth inhibition rate of 0% and absorbance of a region in which cells were not seeded is
defined as a cell growth inhibition rate of 100%, a log conc value in terms of log (inhibition rate/(100-inhibition rate)) and a plotted IC50 value (μM) were calculated. The results are
shown in Table 1 [18]. As a control drug, STI571 was used.
atom in the pyrimidine ring also played an important role in
antitumor activities to some extent. So in our next study, we
will keep these pharmacophores and select 7b, and 7e as
candidate compounds for futher development.
The precipitate was filtrated, washed with ethanol and dried
to afford white powder (14.8 g, 87.3 % yield), mp: >250 oC.
4-methyl-3-(2-methyl-6-morpholinopyrimidin-4-ylamino)
benzoic acid (4b)
The intermediate 4-methyl-3-(2-methyl-6-morpholino-
pyrimidin-4-ylamino) benzoic acid (4b) was prepared from
methyl 3-amino-4-methylbenzoate and 4,6-dichloro-2-
methylpyrimidine by using the general procedure in 4a de-
scribed above. Yield: 89.7 %, mp: >250 °C.
EXPERIMENTAL SECTION
Synthesis of the Intermediates (4a, 4b)
3-(2-methyl-6-morpholinopyrimidin-4-ylamino)benzoic
acid (4a)
Step1: methyl 3-(6-chloro-2-methylpyrimidin-4-ylamino)
benzoate
Synthesis of the Target Compounds
The General Procedure for the Synthesis of N-(substituen-
tedphenyl)-3-(2-methyl-6-morpholinopyrimidin-4-ylamino)
benzamides (5a-5d)
Compound 1a (10.0 g, 66.0 mmol), 4,6-dichloro-2-
methylpyrimidine (13.0 g, 80.0 mmol) and 2 mL con.HCl
(36%) were added to 200 mL isopropanol. When the mixture
was refluxed for 3h, the solution was cooled and placed in
the refrigerator overnight. The precipitate was colleted by
filtration, washed with cold isopropanol (2ꢀ50 mL) to afford
white powder (16.8 g, 92% yield), mp: 192-195 oC.
Compound (4a) (1 mmol) and CDI (1 mmol) were dis-
solved in 15mL of DMSO, then the reaction mixture was
maintained at the temperature of 50 C under nitrogen at-
o
mosphere for 3h. The reaction mixture was cooled to reach
room temperature, substituented phenylamino (1 mmol)
were added and the solution was stirred overnight. The mix-
ture was poured into water, extracted two times with ethyl
acetate, the combined organic layer was dried over anhy-
drous MgSO4. The solvent was removed under vacuum. The
crude product was purified by chromatographic colum to
afford white powder.
Step2: 3-(6-chloro-2-methylpyrimidin-4-ylamino)benzoic
acid (4a)
Methyl 3-(6-chloro-2-methylpyrimidin-4-ylamino) ben-
zoate (15.0 g, 54 mmol) and morpholine (14 mL, 162 mmol)
were dissolved in 1,4-dioxane (150mL). The reaction mix-
ture was held at reflux for 15h, then the solvent was removed
under vaccum affording the expected product (3a). A solu-
tion of 50ml methanol, 30 mL H2O and 4.5 g LiOH was
added to the compound (3a), and the reaction was stirred at
room temperature for 5h. Methanol was evaporated and the
residue was then brought into PH(2~3) with 2 mol/L HCl.
N-p-tolyl-3-(2-methyl-6-morpholinopyrimidin-4-ylamino)-
benzamide (5a)
Yield 68%, mp: 185-188 oC; 1HNMR (DMSO-d6,
400MHz) ꢁ: 2.30 (d, 6H), 3.46 (t, 4H), 3.66 (t, 4H), 5.84 (s,
1H), ꢁ 7.15 (d, 2H), 7.44 (m, 2H), 7.65 (d, 2H), 7.98 (m,

Design, Synthesis and Bioactivities of Phenylamino-Pyrimidine Derivatives
Letters in Drug Design & Discovery, 2011, Vol. 8, No. 7 599
2H), 9.23 (s, 1H), 10.13 (s, 1H); M/z: 404.2 ([M+H]+,
100%). Elem. Anal. Calcd: C, 68.47; H, 6.25; N, 17.36.
Found C, 68.33; H, 6.27; N, 17.39.
N-(4-iodophenyl)-4-methyl-3-(2-methyl-6-morpholinopyri-
midin-4-ylamino)benzamide (6c)
Yield 65%, mp: 193-196 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ: 2.25 (d, 6H), 3.38 (t, 4H), 3.62 (t, 4H), 5.59 (s,
1H), 7.10 (t, 1H), 7.36 (m, 2H), 7.65 (m, 2H), 7.76 (dd, 1H),
8.02 (d, 1H), 8.46 (s, 1H), 10.14 (s, 1H); M/z: 530.1
([M+H]+, 100%). Elem. Anal. Calcd: C, 52.18; H, 4.57; N,
13.23. Found C, 52.23; H, 4.56; N, 13.26.
N-(4-fluorophenyl)-3-(2-methyl-6-morpholinopyrimidin-4-
ylamino)benzamide (5b)
Yield 72%, mp: 191-194 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ: 2.32 (s, 3H), 3.45 (t, 4H), 3.66 (t, 4H), 5.84 (s,
1H), 7.19 (t, 2H), 7.45 (m, 2H), 7.92 (m, 4H), 9.26 (s, 1H),
10.13 (s, 1H); M/z: 408.2 ([M+H]+, 100%). Elem. Anal.
Calcd: C, 64.85; H, 5.44; N, 17.19. Found C, 64.98; H, 5.47;
N, 17.17.
N-(2-chloro-4-methylphenyl)-4-methyl-3-(2-methyl-6-mor-
pholinopyrimidin-4-ylamino)benzamide (6d)
Yield 68%, mp: 188-191 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ: 2.25 (t, 9H), 3.38 (t, 4H), 3.62 (t, 4H), 5.58 (s,
1H), 7.14 (d, 2H), 7.37 (d, 1H), 7.66 (m, 2H), 8.01 (s, 1H),
8.46 (s, 1H), 10.06 (s, 1H); M/z: 452.2 ([M+H]+, 100%).
Elem. Anal. Calcd: C, 63.78; H, 5.80; N, 15.50. Found C,
63.91; H, 5.81; N, 15.48.
N-(4-iodophenyl)-3-(2-methyl-6-morpholinopyrimidin-4-
ylamino)benzamide (5c)
Yield 67%, mp: 197-200 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ: 2.32 (s, 3H), 3.45 (t, 4H), 3.66 (t, 4H), 5.84 (s,
1H), 7.56 (m, 6H), 7.99 (m, 2H), 9.25 (s, 1H), 10.22 (s, 1H);
M/z: 516.1 ([M+H]+, 100%). Elem. Anal. Calcd: C, 51.27; H,
4.30; N, 13.59. Found C, 51.53; H, 4.31; N, 13.55.
The General Procedure for the Synthesis of N-(substituen-
tedphenyl)-3-(5-chloro-2-methyl-6-morpholinopyrimidin-4-
ylamino)-benzamides (7a-7g)
N-(2-chloro-4-methylphenyl)-3-(2-methyl-6-morpholino-
pyrimidin-4-ylamino)benzamide (5d)
Compound (4a) (1 mmol) was dissolved in 20 mL
thionyl chloride and 1 mL DMF, the solution was held at
reflux for 4h, then the thionyl chloride was removed under
vaccum affording the expected acyl chloride. To a suspen-
sion of substituented phenylamino (1 mmol) and Et3N (3.5
mL) in CH2Cl2 (15 mL) was added a solution of the expected
acyl chloride in CH2Cl2 at dropwise with the temperature
Yield 71%, mp: 197-200 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ: 2.30 (d, 6H), 3.45 (t, 4H), 3.65 (t, 4H), 5.84
(s,1H), 7.15 (d, 2H), 7.44 (m, 2H), 7.65 (d, 1H), 7.98 (m,
2H), 9.23 (s, 1H), 10.13 (s, 1H); M/z: 438.2 ([M+H]+,
100%). Elem. Anal. Calcd: C, 63.08; H, 5.52; N, 15.99.
Found C, 63.02; H, 5.54; N, 15.97.
o
under 10 C. The reaction mixture was allowed to warm to
room temperature and stirred overnight. The solution was
poured into 100 mL CH2Cl2, washed with water (2ꢁ100 mL),
saturated sodium carbonate (2ꢁ100 mL) and brine (2ꢁ100
mL). The organic layer was dried over anhydrous MgSO4.
The solvent was removed under vacuum. The crude product
was purified by chromatographic colum to afford white
powder.
The General Procedure for the Synthesis of N-(substituen-
tedphenyl)-4-methyl-3-(2-methyl-6-morpholinopyrimidin-
4-ylamino)-benzamides (6a-6d)
Compound (4b) (1 mmol) and CDI (1 mmol) were dis-
solved in 15mL of DMSO, then the reaction mixture was
o
maintained at the temperature of 50 C under nitrogen at-
mosphere for 3h. The reaction mixture was cooled to reach
room temperature, substituented phenylamino (1 mmol)
were added and the solution was stirred overnight. The mix-
ture was poured into water, extracted two times with ethyl
acetate, the combined organic layer was dried over anhy-
drous MgSO4. The solvent was removed under vacuum,
crude product was purified by chromatographic colum to
afford white powder.
N-p-tolyl-3-(5-chloro-2-methyl-6-morpholinopyrimidin-4-
ylamino)-benzamide (7a)
Yield 74%, mp: 198-201 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ: 2.30 (d, 6H), 3.45 (t, 4H), 3.71 (t, 4H), 7.15 (d,
2H), 7.44 (t, 1H), 7.64 (m, 3H), 7.90 (dd, 1H), 8.16 (t, 1H),
8.84 (s, 1H), 10.12 (s, 1H); M/z: 438.2 ([M+H]+, 100%).
Elem. Anal. Calcd: C, 63.08; H, 5.52; N, 15.99. Found C,
62.89; H, 5.23; N, 15.96.
N-p-tolyl-4-methyl-3-(2-methyl-6-morpholinopyrimidin-4-
ylamino)-benzamides (6a)
N-(4-fluorophenyl)-3-(5-chloro-2-methyl-6-morpholino-
pyrimidin-4-ylamino)- benzamide (7b)
Yield 70%, mp: 179-182 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ: 2.26 (t, 9H), 3.38 (t, 4H), 3.62 (t, 4H), 5.58 (s,
1H), 7.14 (d, 2H), 7.37 (d, 1H), 7.66 (m, 3H), 8.01 (s, 1H),
8.46 (s, 1H), 10.06 (s, 1H); M/z: 418.2 ([M+H]+, 100%).
Elem. Anal. Calcd: C, 69.04; H, 6.52; N, 16.77. Found C,
69.11; H, 6.53; N, 16.79.
Yield 76%, mp: 211-213 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ: 2.50 (s, 3H), 3.44 (t, 4H), 3.71 (t, 4H), 7.20 (t,
2H), 7.46 (t, 1H), 7.62 (d, 1H), 7.85 (m, 3H), 8.18 (s, 1H),
8.89 (s, 1H), 10.30 (s, 1H); M/z: 442.1 ([M+H]+, 100%).
Elem. Anal. Calcd: C, 59.80; H, 4.79; N, 15.85. Found C,
59.86; H, 4.80; N, 15.83.
N-(4-fluorophenyl)-4-methyl-3-(2-methyl-6-morpholino-
pyrimidin-4-ylamino)benzamide (6b)
N-(4-iodophenyl)-3-(5-chloro-2-methyl-6-morpholinopyri-
midin-4-ylamino)- benzamide (7c)
Yield 74%, mp: 188-191 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ: 2.25 (d, 6H), 3.38 (t, 4H), 3.62 (t, 4H), 5.58 (s,
1H), 7.18 (t, 2H), 7.38 (d, 1H), 7.68 (dd, 1H), 7.78 (m, 2H),
8.02 (s, 1H), 8.46 (s, 1H), 10.20 (s, 1H); M/z: 422.2
([M+H]+, 100%). Elem. Anal. Calcd: C, 65.54; H, 5.74; N,
16.62. Found C, 65.93; H, 5.69; N, 16.63.
Yield 70%, mp: 195-198 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ: 2.32 (s, 3H), 3.46 (t, 4H), 3.70 (t, 4H), 7.46 (t,
1H), 7.66 (m, 5H), 7.91 (dd, 1H), 8.18 (s, 1H), 8.86 (s, 1H),
10.30 (s, 1H); M/z: 550.0 ([M+H]+, 100%). Elem. Anal.
Calcd: C, 48.06; H, 3.85; N, 12.74. Found C, 48.20; H, 3.85;
N, 12.77.

600 Letters in Drug Design & Discovery, 2011, Vol. 8, No. 7
Hong et al.
N-(2-chloro-6-methylphenyl)-3-(5-chloro-2-methyl-6-mor-
pholinopyrimidin-4-ylamino)-benzamide (7d)
N-(4-fluorophenyl)-4-methyl-3-(5-chloro-2-methyl-6-mor-
pholinopyrimidin-4-ylamino)-benzamide (8b)
Yield 75%, mp: 198-201 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ:2.30 (d, 6H), 3.45 (t, 4H), 3.70 (t, 4H), 7.15 (d,
2H), 7.45 (m, 1H), 7.64 (m, 3H), 7.90 (dd, 1H), 8.17 (m,
1H), 8.84 (s, 1H), 10.12 (s, 1H); M/z: 472.1 ([M+H]+,
100%). Elem. Anal. Calcd: C, 58.48; H, 4.91; N, 14.83.
Found C, 58.36; H, 4.90; N, 14.82.
Yield 75%, mp: 205-207 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ: 2.19 (d, 6H), 3.43 (t,4H), 3.71 (t, 4H), 7.18 (t,
2H), 7.41 (d, 1H), 7.82 (m, 4H), 8.63 (s, 1H), 10.25 (s, 1H);
M/z: 456.2 ([M+H]+, 100%). Elem. Anal. Calcd: C, 60.59; H,
5.08; N, 15.36. Found C, 60.41; H, 5.09; N, 15.31.
N-(4-iodophenyl)-4-methyl-3-(5-chloro-2-methyl-6-mor-
pholinopyrimidin-4-ylamino)-benzamide (8c)
N-(3-(trifluoromethyl)phenyl)-3-(5-chloro-2-methyl-6-mor-
pholinopyrimidin-4-ylamino)-benzamide (7e)
Yield 71%, mp:194-197 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ: 2.19 (d, 6H), 3.44 (t, 4H), 3.70 (t, 4H), 7.41 (d,
1H), 7.66 (m, 4H), 7.77 (dd, 1H), 7.88 (d, 1H), 8.58 (s, 1H),
10.25 (s, 1H); M/z: 564.1 ([M+H]+, 100%). Elem. Anal.
Calcd: C, 49.00; H, 4.11; N, 12.42. Found C, 49.15; H, 4.10;
N, 12.43.
Yield 69%, mp: 214-217 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ: 2.32 (s, 3H), 3.45 (t, 4H), 3.71(t, 4H), 7.56 (m,
4H), 7.92 (d, 1H), 8.06 (d, 1H), 8.23 (d, 2H), 8.87 (s, 1H),
10.52 (s, 1H); M/z: 492.1 ([M+H]+, 100%). Elem. Anal.
Calcd: C, 56.16; H, 4.30; N, 14.24. Found C, 56.10; H, 4.31;
N, 14.27.
N-(2-chloro-6-methylphenyl)-4-methyl-3-(5-chloro-2-
N-(4-methoxyphenyl)-3-(5-chloro-2-methyl-6-morpholino-
pyrimidin-4-ylamino)- benzamide (7f)
methyl-6-morpholinopyrimidin-4-ylamino)-benzamide (8d)
Yield 73%, mp: 192-195 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ: 2.20 (t, 9H), 3.44 (t, 4H), 3.70 (t, 4H), 7.36 (m,
4H), 7.86 (m, 2H), 8.59 (s, 1H), 9.94 (s, 1H); M/z: 486.1
([M+H]+, 100%). Elem. Anal. Calcd: C, 59.26; H, 5.18; N,
14.40. Found C, 59.32; H, 5.17; N, 14.36.
Yield 78%, mp: 193-196 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ: 2.32 (s, 3H), 3.45 (t, 4H), ꢀ 3.70 (t, 4H), 3.75 (s,
3H), 6.93 (d, 2H), 7.44 (m, 1H), 7.64 (m, 3H), 7.89 (d, 1H),
8.16 (s, 1H), 8.88 (s, 1H), 10.11 (s, 1H); M/z: 454.2
([M+H]+, 100%). Elem. Anal. Calcd: C, 60.86; H, 5.33; N,
15.43. Found C, 61.04; H, 5.32; N, 15.40.
N-(3-(trifluoromethyl)phenyl)-4-methyl-3-(5-chloro-2-
methyl-6-morpholinopyrimidin-4-ylamino)-benzamide (8e)
N-(4-(trifluoromethoxy)phenyl)-3-(5-chloro-2-methyl-6-
morpholinopyrimidin-4-ylamino)-benzamide (7g)
Yield 68%, mp: 212-215 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ: 2.20 (d, 6H), 3.44 (t, 4H), 3.71 (t, 4H), 7.44 (t,
2H), 7.59 (t, 1H), 7.80 (dd, 1H), 7.92 (s, 1H), 8.07 (d, 1H),
8.24 (s, 1H), 8.60 (s, 1H), 10.46 (s, 1H); M/z: 506.2
([M+H]+, 100%). Elem. Anal. Calcd: C, 56.98; H, 4.58; N,
13.84. Found C, 56.80; H, 4.58; N, 13.79.
Yield 67%, mp: 192-195 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ: 2.32 (s, 3H), 3.46 (t, 4H), 3.70 (t, 4H), 7.43 (m,
3H), 7.63 (d, 1H), 7.91 (m, 3H), 8.20 (s, 1H), 8.87 (s, 1H),
10.40 (s, 1H); M/z: 508.1 ([M+H]+, 100%). Elem. Anal.
Calcd: C, 54.39; H, 4.17; N, 13.79. Found C, 54.23; H, 4.17;
N, 13.83.
N-(4-methoxyphenyl)-4-methyl-3-(5-chloro-2-methyl-6-
morpholinopyrimidin-4-ylamino)-benzamide (8f)
The General Procedure for the Synthesis of N-(substituen-
tedphenyl)-4-methyl-3-(5-chloro-2-methyl-6-morpholinopy-
rimidin-4-ylamino)-benzamide (8a-8g)
Yield 76%, mp: 186-189 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ: 2.18 (d, 6H), 3.43 (t, 4H), 3.71(t, 4H), 6.92 (d,
2H), 7.40 (d, 1H), 7.76 (m, 4H), 8.62 (s, 1H), 10.07 (s, 1H);
M/z: 468.2 ([M+H]+, 100%). Elem. Anal. Calcd: C, 61.60; H,
5.60; N, 14.97. Found C, 61.41; H, 5.61; N, 15.01.
Compound (4b) (1 mmol) was dissolved in 20mL thionyl
chloride and 1mL DMF, the solution was held at reflux for
4h, then the thionyl chloride was removed under vaccum
affording the expected acyl chloride. To a suspension of sub-
stituented phenylamino (1 mmol) and Et3N (3.5 mL) in
CH2Cl2 (15 mL) was added a solution of the expected acyl
chloride in CH2Cl2 at dropwise with the temperature under
N-(4-(trifluoromethoxy)phenyl)-4-methyl-3-(5-chloro-2-
methyl-6-morpholinopyrimidin-4-ylamino)-benzamide (8g)
Yield 69%, mp: 187-190 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ: 2.19 (d, 6H), 3.45 (t, 4H), 3.70 (t, 4H), 7.39 (m,
3H), 7.84 (m, 4H), 8.59 (s, 1H), 10.35 (s, 1H); M/z: 522.1
([M+H]+, 100%). Elem. Anal. Calcd: C, 55.23; H, 4.44; N,
13.42. Found C, 55.17; H, 4.43; N, 13.43.
o
10 C. The reaction mixture was allowed to warm to room
temperature and stirred overnight. The solution was poured
into 100 mL CH2Cl2, washed with water (2ꢁ100 mL), satu-
rated sodium carbonate (2ꢁ100 mL) and brine (2ꢁ100 mL).
The organic layer was dried over anhydrous MgSO4. The
solvent was removed under vacuum. The crude product was
purified by chromatographic colum to afford white powder.
ACKNOWLEDGEMENTS
This work was financially supported by the National
Natural Science Foundation of China (Grant No.30973616).
N-p-tolyl-4-methyl-3-(5-chloro-2-methyl-6-morpholinopyri-
midin-4-ylamino)- benzamide (8a)
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Yield 72%, mp: 190-193 oC; 1HNMR (DMSO-d6,
400MHz) ꢀ: 2.19 (t, 9H), 3.44 (t, 4H), 3.70 (t, 4H), 7.14 (d,
2H), 7.39 (d, 1H), 7.76 (m, 4H), 8.57 (s, 1H), 10.40 (s, 1H);
M/z: 452.2 ([M+H]+, 100%). Elem. Anal. Calcd: C, 63.78; H,
5.80; N, 15.50. Found C, 63.72; H, 5.81; N, 15.52.
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