Synthesis and cytotoxic activity of diaryl urea derivatives with a 4-methylpiperazinylcarbonyl moiety

Article abstract of DOI:10.1007/s00044-012-0398-y

Eleven diarylurea derivatives (1a-1k) bearing N-methylpiperazinyl moiety were synthesized by the reaction of isocyanate with arylamine. The structures of 1a-1k were characterized by ¹H-NMR and MS. The cytotoxic activities of 1a-1k were evaluated via MTT method against human lung adenocarcinoma epithelial cell line A549 and human prostate carcinoma cell line PC3. Compounds 1d and 1k displayed potent cytotoxic activity. The results suggested that the activities are considerably related to the substituent group at another phenyl ring.

Full text of DOI:10.1007/s00044-012-0398-y

MEDICINAL
CHEMISTRY
RESEARCH
Med Chem Res (2013) 22:3857–3862
DOI 10.1007/s00044-012-0398-y
ORIGINAL RESEARCH
Synthesis and cytotoxic activity of diaryl urea derivatives
with a 4-methylpiperazinylcarbonyl moiety
•
•
•
Wei Xuan Wen Ding Hong-xiang Hui
San-qi Zhang
Received: 7 June 2012 / Accepted: 6 December 2012 / Published online: 13 December 2012
Ó Springer Science+Business Media New York 2012
Abstract Eleven diarylurea derivatives (1a–1k) bearing
N-methylpiperazinyl moiety were synthesized by the
reaction of isocyanate with arylamine. The structures of
1a–1k were characterized by 1H-NMR and MS. The
cytotoxic activities of 1a–1k were evaluated via MTT
method against human lung adenocarcinoma epithelial cell
line A549 and human prostate carcinoma cell line PC3.
Compounds 1d and 1k displayed potent cytotoxic activity.
The results suggested that the activities are considerably
related to the substituent group at another phenyl ring.
(Yu et al., 2012), antimalarial (Madapa et al., 2009), anti-
diabetic (Anandan and Gless, 2010), analgesic (Pacchiano
et al., 2011), and weed activities (Beaver et al., 1957). In
recent years, some kinase inhibitors with aryl urea structure
have been launched for the treatment of cancers or tested
clinically. For example, linifanib, developed by Abbott, is
used for the treatment of colorectal cancer (Ji et al., 2008).
Sorafenib, developed by Bayer, is used for the treatment of
advanced renal cell carcinoma (Liu et al., 2006). Tandutinib
is used to treat kidney cancer and acute leukemia (Cheng
and Paz, 2008). Meanwhile, PKI-179 (Venkatesan et al.
2010a), PKI-587 (Venkatesan et al. 2010b), and PKI-402
(Dehnhardt et al., 2009), developed by Wyeth, were iden-
tified as drug candidates which are now in clinical devel-
opment for the treatment of solid tumors. Now, medicinal
chemists have directed considerable attention on the anti-
tumor activity of aryl urea derivative.
Keywords Diaryl urea ꢀ Synthesis ꢀ Cytotoxic activity
Introduction
Aryl urea moiety is present in many classes of biologically
active compounds (Denoyelle et al., 2012; Marvania et al.,
2011; Sonmez et al., 2011; Witherington et al., 2006; Pan
et al., 2006; Receveur et al., 2004; Ogita et al., 2002;
Anandan et al., 2011). The aryl urea derivatives exhibit a
broad spectrum of biological activities such as antitumor
N-methylpiperazinyl moiety is versatile and privileged
in drug discovery (Philip and Bernard, 1971; Hawes et al.,
1984; Hayakawa et al., 1986). It is known that the
N-methylpiperazine is considered as a reactive functional
group and water soluble moiety. The water solubility and
pharmacokinetic property of a drug could be significantly
improved after N-methylpiperazinyl moiety was linked
(Venkatesan et al., 2010a; Venkatesan et al., 2010b;
Dehnhardt et al., 2009).
In an attempt to develop novel antitumor agents, we try
to introduce N-methylpiperazinyl moiety to the diaryl urea
scaffold to design and synthesize the analogs of sorafenib.
Herein, the N-methylpicolinamide moiety in sorafenib was
replaced by N-methylpiperazinyl, and the oxygen atom in
ether bond was replaced by a carbonyl (Fig. 1). Compared
with sorafenib, the designed compounds showed similar
pharmacophore. Here, we report the synthesis and cyto-
toxic activity of eleven new diarylureas.
W. Xuan ꢀ S. Zhang (&)
Department of Pharmacy, School of Medicine, Xi’an Jiaotong
University, Xi0an 710061, People’s Republic of China
e-mail: sqzhang@mail.xjtu.edu.cn; sqzhang@xjtu.edu.cn
W. Ding ꢀ H. Hui
Institute of Biotherapy, School of Biotechnology,
Southern Medical University, Guangzhou 510515,
People’s Republic of China
123

3858
Med Chem Res (2013) 22:3857–3862
and integral values. Aromatic ring protons were seen at
6.13–7.69 ppm. The NH protons (in the urea) were seen at
about 7.98–8.94 ppm. The 1-methylpiperazinyl protons appe-
ared at about 3.75, 2.48, and 2.36 ppm, respectively.
Cytotoxic assays
All the newly synthesized compounds 1a–1k were evalu-
ated for their cytotoxic activities in vitro via MTT assay
against two human cancer cell lines, including PC-3
(human prostate carcinoma cell line) and A549 (human
lung adenocarcinoma epithelial cell line) at the concen-
tration of 100 lM. Sorafenib was used as a positive con-
trol. The results are summarized in Table 1. Data in the
Table 1 show that most of the synthesized compounds
exhibited weak cytotoxic activity against the two human
cancer cell lines. However, compound 1d displayed dis-
tinct cytotoxic activity with inhibition percentages higher
than 70 and 80 % against A549 and PC3, respectively. It is
worthy to point out that significant inhibition was achieved
for compound 1k with an inhibition percentage higher than
90 % against A549. Compounds 1a only displayed an
inhibition percentage less than 50 % against A549 and
PC3. These data indicate that the activities are considerably
related to the substituent group at another phenyl ring.
To further evaluate cytotoxic activity of compounds 1d
and 1k, two compounds and sorafenib were tested against
A549 over a range of concentrations from 100 lM to
0.01 lM, and the calculated IC50 values were 10.8, 0.57,
and 6.08 lM, respectively. The concentration–response
curve of compounds 1d, 1k, and sorafenib against A549 are
shown in Fig. 2. Compound 1d exhibited cytotoxic activity
close to sorafenib. Compound 1k showed more potent
activity than sorafenib, and both showed significantly
concentration-dependent manner.
Fig. 1 The structure of sorafenib and compound 1
Results and discussion
Synthesis
The synthetic route was outlined in Scheme 1.
The commercially available methyl 4-formylbenzoate 2
was oxidized into 4-(methoxycarbonyl)benzoic acid 3 via
Jones reagent oxidation. The reaction of 3 with oxalyl
chloride in dry methylene chloride gave corresponding acid
chloride, which reacted with 1-methylpiperazine and
afforded 4. The obtained 4 was selectively hydrolyzed and
subsequently acidified to produce 5. Compound 5 is an
amino acid and difficult to purify. So crude product of 5
reacted with thionyl chloride to give acid chloride, which
reacted with sodium azide and afforded acyl azide 6.
Heating the acyl azide 6 in toluene produced isocyanate,
which was not isolated and reacted with anilines to produce
targeted compounds 1a–1k.
All the newly synthesized compounds gave satisfactory
analyses for the proposed structures, which were characterized
on the basis of their 1H-NMR and MS. In 1H-NMR spectra, all
protons were seen according to the expected chemical shift
Scheme 1 Synthesis of target
compounds 1a–1k
123

Med Chem Res (2013) 22:3857–3862
3859
Table 1 The structure and inhibition values of the target compounds
Compd
Ar
% Inhibitiona
A549
PC3
1a
1b
1c
1d
1e
1f
3-ethynylphenyl
3-C14-FPh
45.4
28.4
21.2
74.0
43.1
28.6
22.6
27.5
36.5
19.7
93.5
49.3
29.9
37.1
82.7
27.6
40.0
25.5
35.1
28.0
31.1
ntb
3-CF3Ph
3-Cl-4-(3-FBnO)Ph
3,4-diFPh
3,5-diMeOPh
3-CF34-ClPh
1g
1h
1i
2-Me-3-QPh
3,4-diClPh
1j
3-(piperidine-1-carbonyl)Ph
1k
Sorafenib
–
94.8
91.4
a
Values are means of six experiments
b
Not tested
fragment in drug design. In addition, the activity of benzo-
thiazole derivatives bearing an arylurea moiety at C-6 is
rarely reported. Compound 1k possesses an arylurea moiety
at C-6 and displayed potent cytotoxic activity against A549.
Conclusions
Eleven diarylurea derivatives bearing N-methylpiperazinyl
moiety were synthesized and their cytotoxic activities were
evaluated via MTT method against A549 and PC3. Com-
pounds 1d and 1k displayed potent cytotoxic activity. The
results suggested that the activities are considerably related
to the substituent group at another phenyl ring.
Fig. 2 The concentration–response inhibitory curve of compounds
1d, 1k, and sorafenib against A549 (n = 6)
Experimental
Previous studies have indicated that benzothiazole
derivatives exhibit potent cytotoxic and antitumor agents
(Yoshida et al., 2005; Lion et al., 2006). Benzothiazole
derivatives bearing at C-2 arylurea moiety also showed
interesting cytotoxic activities (Caputo, et al., 2012). There
are two hydrogen-bond acceptors and one hydrogen-bond
donor in the 2-acetylaminobenzothiazole. Thus it is a desired
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
123

3860
Med Chem Res (2013) 22:3857–3862
melting points were determined on a Beijing micromelting-
point apparatus and thermometer was uncorrected.
1H-NMR spectra were recorded on a 300 MHz or 400
Bruker NMR spectrometer with tetramethylsilane (TMS)
as an internal reference. All chemical shifts are reported in
parts per million (ppm). High-resolution exact mass mea-
surements were performed using electrospray ionization
(positive mode) on a quadrupole time-of-flight (QTOF)
mass spectrometer (Maxis Q-TOF, Bruker Inc.).
and stirred at room temperature for 2 h. Then the mixture
was concentrated under reduced pressure. Water (50 ml)
was added to the mixture, which was then acidified with
concentrated hydrochloric acid to pH 2. The aqueous phase
was concentrated to dry under reduced pressure to afford
the crude 4-(1-methylpiperazine-4-carbonyl) benzoic acid
(containing sodium chloride) which was used for the next
step without further isolation.
4-(4-methylpiperazine-1-carbonyl)benzoylazide (6)
4-methoxycarbonylbenzoic acid (3)
A mixture of crude product of 5 (3.95 g, containing sodium
chloride), thionyl chloride (10 ml), toluene (10 ml), and
DMF (3 drops) was refluxed under stirring for 1 h. The
solvent was removed under reduced pressure to give a pale
yellow solid. The solid was dissolved in dry THF (40 ml)
and added dropwise to a suspension of sodium azide
(3.70 g, 56.0 mmol) in 50 ml of THF/water (4:1,V/V)
cooled to 0–5 °C. After addition, the mixture was stirred at
room temperature for 2 h and then concentrated under
reduced pressure. The residue was extracted with ethyl
acetate (50 ml 9 3). The organic layers were combined,
washed with brine (40 ml), dried (MgSO4), filtered, and
concentrated under reduced pressure to afford 2.70 g of a
yellow solid, yield 74.0 % (two steps). mp 103.0–104.0 °C.
HRMS (ESI): calcd. for C13H15N5O2 274.1304 [M ? H]?,
To a stirred solution of aldehyde 2 (16.40 g, 0.1 mol) in
acetone (120 ml) at 0 °C, dropwise freshly prepared Jones
reagent was added until an orange color persisted. [Jones
reagent was prepared by dissolving chromium trioxide
(20.0 g) in 75.0 ml of distilled water and adding 17.0 ml of
concentrated sulfuric acid.] The reaction mixture was
concentrated under reduced pressure. The residue was fil-
tered with suction. The solid was washed with water
(100 ml), dried to afford 3 17.5 g, yield 97.0 %. mp
1
220.0–221.0 °C. H-NMR (400 MHz, DMSO-d6) d: 8.07
(b, 4H, 4 9 Ar–H), 3.89 (s, 3H, O–CH3).
Methyl 4-(4-methylpiperazine-1-carbonyl)benzoate (4)
Oxalyl chloride (5.0 ml, 56.0 mmol, 2.0 eq.) was slowly
added to the mixture of 4-(methoxycarbonyl)benzoic acid 3
(5.00 g, 28.0 mmol) and dried dichloromethane (40 ml).
Then DMF (0.2 ml) was added and the mixture was stirred
at room temperature for 2 h. Meanwhile the reaction mix-
ture changed from a suspension into a clear solution. The
solvent was removed under reduced pressure to give a white
solid. The solid was dissolved in dried THF (40 ml). The
obtained solution was added dropwise to the solution of
1-methylpiperazine (3.7 ml, 34.0 mmol, 1.2 eq.) in dried
THF (40 ml) cooled to about 0 °C. The resulting mixture
was stirred at room temperature for 3 h, concentrated under
reduced pressure. The residue was diluted with water
(40 ml) and extracted with ethyl acetate (50 ml 9 3). The
organic layers were combined, washed with brine (40 ml),
dried (MgSO4), filtered, and concentrated under reduced
pressure to afford 6.86 g of a yellow solid; yield 94.2 %. mp
83.0–84.0 °C. MS m/z 262.1(M?). 1H-NMR (400 MHz,
CDCl3): d 8.08 (d, 2H, Ar–H, J = 8.0 Hz), 7.47 (d, 2H, Ar–
H, J = 8.0 Hz), 3.94 (s, 3H, O–CH3), 3.83 (s, 2H, CON–
CH2), 3.41 (s, 2H, CON–CH2), 2.52 (s, 2H, N–CH2), 2.36
(s, 2H, N–CH2), 2.34 (s, 2H, N–CH3).
1
found 274.1307. H-NMR (300 MHz, CDCl3): d 8.08 (d,
2H, Ar–H, J = 8.0 Hz), 7.48 (d, 2H, Ar–H, J = 8.0 Hz),
3.84 (s, 2H, CON-CH2), 3.40 (s, 2H, CON–CH2), 2.53 (s,
2H, N–CH2), 2.35 (s, 5H, N–CH2, N–CH3).
Synthesis of diaryl urea derivatives 1a–1k
General procedure
The compound 6 (0.27 g, 0.99 mmol) was refluxed in
toluene (5.0 ml) for 1 h under N2 and then cooled to room
temperature. Arylamine was added directly, and the mix-
ture was stirred at room temperature for another 2 h. The
solvent was removed and flash chromatograph of the res-
idue over silica gel, using chloroform/methanol (10:1),
gave 1a–1k as white solids.
1-(3-ethynylphenyl)-3-(4-(4-methylpiperazine-1-carbonyl)-
phenyl)urea (1a) 0.35 g, yield 97.0 %. mp 168.0 *
170.0 °C. MS (m/z) 362.2 (M?). 1H-NMR (400 MHz,
DMSO-d6): d 8.96 (s, 1H, N–H), 8.87 (s, 1H, N–H), 7.67
(s, 1H, Ar–H), 7.52 (d, 2H, Ar–H, J = 8.8 Hz), 7.42 (d,
1H, Ar–H, J = 7.2 Hz), 7.34 (d, 2H, Ar–H, J = 8.8 Hz),
7.30 (m, 1H, Ar–H), 7.10 (d, 1H, Ar–H, J = 7.6 Hz), 4.18
(s, 1H, C:CH), 3.49 (b, 4H, 2 9 CON–CH2), 2.31 (b, 4H,
2 9 N–CH2), 2.20 (s, 3H, N–CH3).
4-(4-methylpiperazine-1-carbonyl)benzoic acid (5)
A solution of NaOH (2 M, 15.0 ml, 30.0 mmol) was added
to a solution of 4 (6.95 g, 26.0 mmol) in MeOH (30 ml)
123

Med Chem Res (2013) 22:3857–3862
3861
1-(3-chloro-4-fluorophenyl)-3-(4-(4-methylpiperazine-1-car-
bonyl)phenyl)urea (1b) 0.38 g, yield 97.0 %. mp 169.5 *
172.5 °C. MS (m/z) 390.1 (M?). 1H-NMR (400 MHz,
CDCl3): d 8.32 (s, 1H, N–H), 8.17 (s, 1H, N–H), 7.47 (d, 1H,
Ar–H, J = 8.4 Hz), 7.40 (s, 1H, Ar–H), 7.22 (d, 2H, Ar–H,
J = 8.8 Hz), 7.14 (s, 1H, Ar–H, J = 8.4 Hz), 7.06 (d, 2H,
Ar–H, J = 8.8 Hz), 3.84 (s, 2H, CON–CH2), 3.48 (s, 2H,
CON–CH2), 2.52 (s, 2H, CON–CH2), 2.47 (s, 5H, N–CH2,
N–CH3).
J = 8.7 Hz), 7.39 (d, 1H, Ar–H, J = 8.7 Hz), 7.20 (d, 2H,
Ar–H, J = 8.1 Hz), 7.06 (d, 2H, Ar–H, J = 8.1 Hz), 3.75
(b, 4H, 2 9 CON–CH2), 2.53 (b, 4H, 2 9 N–CH2), 2.38 (s,
3H, N–CH3).
1-(4-chloro-3-methylphenyl)-3-(4-(4-methylpiperazine-1-
carbonyl)phenyl)urea (1h) 0.28 g, yield 73.0 %, mp
1
166.3 * 168.0 °C. MS (m/z) 386.2(M?). H-NMR (300
MHz, CDCl3): d 8.26 (s, 1H, N–H), 7.52 (s, 1H, N–H), 7.62
(d, 1H, Ar–H), 7.30 (m, 1H, Ar–H), 7.21 (d, 2H, Ar–H), 7.12
(b, 3H, Ar–H), 3.73 (b, 4H, 2 9 CON–CH2), 2.51 (b, 4H,
2 9 N–CH2), 2.38 (s, 3H, N–CH3), 2.33 (s, 3H, Ar–CH3).
1-(4-(4-methylpiperazine-1-carbonyl)phenyl)-3-(3-(trifluo-
romethyl)phenyl)urea (1c) 0.36 g, yield 90.0 %, mp
181.5 * 183 °C. MS (m/z) 406.2(M?). 1H-NMR (400
MHz, CDCl3): d 8.58 (s, 1H, N–H), 8.17 (s, 1H, N–H), 7.69
(s, 1H, Ar–H), 7.62 (d, 1H, Ar–H, J = 8.0 Hz), 7.38 (t, 1H,
Ar–H), 7.27 (s, 1H, Ar–H), 7.22 (d, 2H, Ar–H,
J = 8.4 Hz), 7.08 (d, 2H, Ar–H, J = 8.4 Hz), 3.84 (s, 2H,
CON–CH2), 3.48 (s, 2H, CON–CH2), 2.51 (b, 2H, N–CH2),
2.40 (s, 5H, N–CH2, N–CH3).
1-(3, 4-dichlorophenyl)-3-(4-(4-methylpiperazine-1-carbo-
nyl)phenyl)urea (1i) 0.34 g, yield 83.0 %, mp 196.0 *
198.0.0 °C. MS (m/z) 406.1(M?). 1H-NMR (300 MHz,
CDCl3): d 8.42 (s, 1H, N–H), 8.04 (s, 1H, N–H), 7.57 (s,
1H, Ar–H), 7.32 (d, 1H, Ar–H, J = 8.1 Hz), 7.30 (d, 1H,
Ar–H, J = 8.1 Hz), 7.20 (d, 2H, Ar–H, J = 7.8 Hz), 7.04
(d, 2H, Ar–H, J = 7.8 Hz), 3.75 (b, 4H, 2 9 CON–CH2),
2.48 (b, 4H, 2 9 N–CH2), 2.36 (s, 3H, N–CH3).
1-(4-(3-fluorobenzyloxy)-3-chlorophenyl)-3-(4-(4-methyl-
piperazine-1-carbonyl)phenyl)urea (1d) 0.43 g, yield
1
86.0 %, mp 193.5 * 194.5 °C. MS (m/z) 496.2(M?). H-
1-(4-(4-methylpiperazine-1-carbonyl)phenyl)-3-(4-(piperi-
dine-1-carbonyl)phenyl)urea (1j) 0.37 g, yield 83.0 %,
mp 110.2 * 111.8 °C. MS (m/z) 449.2(M?). 1H-NMR
(300 MHz, CDCl3): d 8.47 (s, 1H, N–H), 8.31 (s, 1H, N–
H), 7.41 (d, 2H, Ar–H), 7.28–7.33 (m, 4H, Ar–H), 7.21 (d,
1H, Ar–H), 6.94 (d, 1H, Ar–H), 3.39–3.73 (m, 8H,
4 9 CON–CH2), 2.45 (b, 4H, 2 9 N–CH2), 2.33 (s, 3H,
N–CH3), 1.70 (m, 4H, 2 9 CH2), 1.55 (m, 2H, CH2).
NMR (300 MHz, CDCl3): d 8.17 (s, 1H, N–H), 8.14 (s, 1H,
N–H), 7.37 (s, 1H, Ar–H), 7.32 (d, 1H, Ar–H), 7.17–7.23
(m, 5H, Ar–H), 7.06 (d, 2H, Ar–H), 6.99 (d, 1H, Ar–H),
6.81 (d, 1H, Ar–H), 5.04 (s, 2H, ArCH2–O), 3.75 (m, 4H,
2 9 CON–CH2), 2.44 (b, 4H, 2 9 N–CH2), 2.32 (s, 3H,
N–CH3).
1-(3, 4-difluorophenyl)-3-(4-(4-methylpiperazine-1-carbo-
nyl)phenyl)urea (1e) 0.35 g, yield 93.0 %, mp 190.0 *
190.5 °C. MS (m/z) 374.2(M?). 1H-NMR (300 MHz,
CDCl3): d 8.31 (s, 1H, N–H), 7.96 (s, 1H, N–H), 7.49 (s,
1H, Ar–H), 7.32 (d, 1H, Ar–H, J = 7.8 Hz), 7.30 (d, 1H,
Ar–H, J = 7.8 Hz), 7.20 (d, 2H, Ar–H, J = 8.1 Hz), 7.04
(d, 2H, Ar–H, J = 8.1 Hz), 3.75 (b, 4H, 2 9 CON–CH2),
2.48 (b, 4H, 2 9 N–CH2), 2.36 (s, 3H, N–CH3).
1-(2-acetamidobenzo[d]thiazol-5-yl)-3-(4-(4-methylpiper-
azine-1-carbonyl)phenyl) urea (1k) 0.26 g, yield 58.3 %,
mp [ 220 °C. HRMS (ESI): calcd. for C22H25N6O3S
453.1709 [M ? H]?, found 453.1700. 1H-NMR (400 MHz,
DMSO-d6): d 12.26 (s, 1H, N–H), 8.94 (s, 1H, N–H), 8.88
(s, 1H, N–H), 8.12 (s, 1H, Ar–H), 7.65 (d, 1H, Ar–H,
J = 8.8 Hz), 7.53 (d, 2H, Ar–H, J = 8.4 Hz), 7.43 (d, 1H,
Ar–H, J = 8.8 Hz), 7.34 (d, 2H, Ar–H, J = 8.4 Hz), 3.49
(b, 4H, 2 9 CON–CH2), 2.32 (b, 4H, 2 9 N–CH2), 2.20
(s, 3H, N-CH3), 2.19 (s, 3H, COCH3).
1-(3, 5-dimethoxyphenyl)-3-(4-(4-methylpiperazine-1-car-
bonyl)phenyl)urea (1f) 0.28 g, yield 70.0 %, mp 194.0 *
196.0 °C. MS (m/z) 398.2(M?).1H-NMR (300 MHz,
CDCl3): d 8.16 (s, 1H, N–H), 8.11 (s, 1H, N–H), 7.21 (b,
2H, Ar–H), 7.12 (b, 2H, Ar–H), 6.66 (s, 2H, Ar–H), 6.17 (s,
2H, Ar–H), 3.76 (s, 3H, O–CH3), 3.75 (s, 3H, O–CH3), 3.56
(b, 4H, 2 9 CON–CH2), 2.48 (b, 4H, 2 9 N–CH2), 2.35 (s,
3H, N–CH3).
Cytotoxic activity
Cellular chemosensitivity was determined by a modified
MTT (3-[4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tet-
razolium bromide) method assay in vitro. In brief, A549 or
PC3 cells in 100 ll culture medium were seeded into
96-well microplates, respectively, and incubated at 37 °C
for 24 h prior to drug exposure. Cell numbers were titrated
to keep control cells growing in the exponential phase
throughout the 48 h incubation period. Cells were treated
1-(4-chloro-3-(trifluoromethyl)phenyl)-3-(4-(4-methylpi-
perazine-1-carbonyl)phenyl)urea (1g) 0.40 g, yield
92.0 %, mp 212.5.0 * 214.0 °C. MS (m/z) 440.1(M?).
1H-NMR (300 MHz, CDCl3): d 8.66 (s, 1H, N–H), 7.97 (s,
1H, N–H), 7.75 (s, 1H, Ar–H), 7.66 (d, 1H, Ar–H,
123

3862
Med Chem Res (2013) 22:3857–3862
Lion CJ, Matthews CS, Wells G, Bradshaw TD, Stevens MFG,
Westwell AD (2006) Antitumour properties of fluorinated
benzothiazole-substituted hydroxycyclohexa-2, 5-dienones. Bio-
org Med Chem Lett 16:5005–5008
Liu L, Cao YC, Chen C, Zhang XM, McNabola A, Wilkie D,
Wilhelm S, Lynch M, Carter C (2006) Sorafenib Blocks the
RAF/MEK/ERK Pathway, Inhibits Tumor Angiogenesis, and
Induces Tumor Cell Apoptosis in Hepatocellular Carcinoma
Model PLC/PRF/5. Cancer Res 66:11851–11858
Madapa S, Tusi Z, Mishra A, Srivastava K, Pandey SK, Tripathi R,
Puri SK, Batra S (2009) Search for new pharmacophores for
antimalarial activity. Part II: synthesis and antimalarial activity
of new 6-ureido-4-anilinoquinazolines. Bioorg Med Chem
17:222–234
with final concentrations of 100.0, 10.0, 1.0, 0.1, and
0.01 lM of tested compounds simultaneously and incu-
bated for 48 h and then 50 ll of MTT solution (5 mg/ml in
medium) was added to each well and incubated for 2.5 h.
The formed blue formazan crystals were pelleted to the
bottom of the well by centrifugation, separated from the
supernatant, and dissolved in 150 ll of DMSO. The optical
density at 570 nm was determined by an ELISA reader.
Two separate experiments with triplicate data were per-
formed to obtain mean cell viability. The IC50 values were
calculated according to the inhibition ratios.
Marvania B, Lee PC, Chaniyara R, Dong HJ, Suman S, Kakadiya R,
Chou TC, Lee TC, Shah A, Su TL (2011) Design, synthesis and
antitumor evaluation of phenyl N-mustard-quinazoline conju-
gates. Bioorg Med Chem 19:1987–1998
Acknowledgments Financial support from The National Natural
Science Foundation of China (Grant No. 21072156) is gratefully
acknowledged.
Ogita H, Isobe Y, Takaku H, Sekine R, Goto Y, Misawa S, Hayashi H
(2002) Synthesis and structure–activity relationship of diaryla-
mide derivatives as selective inhibitors of the proliferation of
human endothelial cells. Bioorg Med Chem 10:3473–3480
Pacchiano F, Carta F, Vullo D (2011) Inhibition of b-carbonic
anhydrases with ureido-substituted benzenesulfonamides. Bioorg
Med Chem Lett 21:102–105
Pan WT, Miao HQ, Xu YJ, Navarro EC, Tonra JR, Erik C, Armin L, Paul
K, Alexander SK, Wai CW, Liu H (2006) 1-[4-(1H-Benzoimidazol-
2-yl)-phenyl]-3-[4-(1H-benzoimidazol-2-yl)-phenyl]urea derivatives
as small molecule heparanase inhibitors. Bioorg Med Chem Lett
16:409–412
References
Anandan SK, Gless RD (2010) Exploration of secondary and tertiary
pharmacophores in unsymmetrical N, N-diaryl urea inhibitors of
soluble epoxide hydrolase. Bioorg Med Chem Lett 20:
2740–2744
Anandan SK, Webb HK, Chen D, Wang YX, Aavula BR, Cases S,
Cheng Y, Do ZN, Mehra U, Tran V, Vincelette J, Waszczuk J,
White K, Kenneth RW, Zhang LN, Paul DJ, Bruce DH, Dinesh
VP, Randall WD, Euan M, Sabry J, Gless R (2011) 1-(1-Acetyl-
piperidin-4-yl)-3-adamantan-1-yl-urea (AR9281) as a potent,
selective, and orally available soluble epoxide hydrolase inhib-
itor with efficacy in rodent models of hypertension and
dysglycemia. Bioorg Med Chem Lett 21:983–988
Beaver DJ, Roman DP, Stoffel PJ (1957) The preparation and
bacteriostatic activity of substituted ureas. J Am Chem Soc
79:1236–1245
Caputo R, Calabro ML, Micale N, Schimmer AD, Ali M, Zappala M,
Grasso S, Med Chem Res (2012) Synthesis of benzothiazole
derivatives and their biological evaluation as anticancer agents.
Med Chem Res 21:2644–2651
Philip MG, Bernard M (1971) Morphogenesis derivatives on vaccinia
virus and other rifamycin similar effect of rifampin. J Virol
8:225–231
Receveur JM, Emelie B, Trond U, Paul BL, Pia
K (2004)
4-Acylamino- and 4-ureidobenzamides as melanin-concentrating
hormone (MCH) receptor 1 antagonists. Bioorg Med Chem Lett
14:5075–5080
Sonmez F, Sevmezler S, Atahan A, Ceylan M, Demir D, Gencer N,
Arslan O, Kucukislamoglu
M (2011) Evaluation of new
chalcone derivatives as polyphenol oxidase inhibitors. Bioorg
Med Chem Lett 21:7479–7482
Venkatesan AM, Chen Z, Santos OD (2010a) PKI-179: an orally
efficacious dual phosphatidylinositol-3-kinase (PI3 K)/mamma-
lian target of rapamycin (mTOR) inhibitor. Bioorg Med Chem
Lett 20:5869–5873
Cheng Y, Paz K (2008) Tandutinib, an oral, small molecule inhibitor
of FLT3 for the treatment of AML andother cancer indications.
Idrugs 11:46–56
Dehnhardt CM, Venkatesan AM, Delos SE (2009) Lead optimization
of N-3-Substituted 7-morpholinotriazolopyrimidines as dual
phosphoinositide 3-kinase/mammalian target of rapamycin
inhibitors: discovery of PKI-402. J Med Chem 53:798–810
Venkatesan AM, Dehnhardt CM, Delos Santos E (2010b) Bis(mor-
pholino-1,3,5-triazine) derivatives: potent adenosine 50-triphos-
phate competitive phosphatidylinositol-3-kinase/mammalian
target of rapamycin inhibitors: discovery of compound 26
(PKI-587), a highly efficacious dual inhibitor. J Med Chem 53:
2636–2645
Witherington J, Blaney EL, Bordas V, Elliott RL, Gaiba A, Garton N,
Green PM, Naylor A, Smith DG, Spalding DJ, Takl AK, Ward
RW (2006) Pyridone derivatives as potent, orally bioavailable
VLA-4 integrin antagonists. Bioorg Med Chem Lett 16:
5538–5541
Yoshida M, Hayakawa I, Hayashi N, Agatsuma T, Oda Y, Tanzawa F,
Iwasaki S, Koyama K, Furukawa H, Kurakatad S, Suganob Y
(2005) Synthesis and biological evaluation of benzothiazole
derivatives as potent antitumor agents. Bioorg Med Chem Lett
15:3328–3332
´
Denoyelle S, Chen T, Chen L, Wang YB, Klosi E, Jose AH, Bertal
HA, Michael C (2012) In vitro inhibition of translation initiation
by N, N-diarylureas-potential anti-cancer agents. Bioorg Med
Chem Lett 22:402–409
Hawes EM, Shetty HU, Cooper JK, Rauw G, McKay G, Midha KK
(1984) Radioimmunoassay for psychotropic drugs III: synthesis
and properties of haptens for trifluoperazine and fluphenazine.
J Pharm Sci 73:247–250
Hayakawa I, Atarashi S, Yokohama S, Imamura M, Sakano K,
Furukawa M (1986) Synthesis and antibacterial activities of
optically active ofloxacin. Antimicrob Agents Chemother 29:
163–164
Ji ZQ, Asma AA, Daniel HA, Jennifer JB, Peter FB, George AC,
Gilbert D, Keith BG, Guo J, Christopher MH, Li JL, Patrick AM
(2008) 3-Amino-benzo[d]isoxazoles as novel multitargeted
inhibitors of receptor tyrosine kinases. J Med Chem 51:
1231–1241
Yu B, Tang LD, Li YL, Song SH, Ji XL, Lin MS, Wu CF (2012)
Design, synthesis and antitumor activity of 4-aminoquinazoline
derivatives targeting VEGFR-2 tyrosine kinase. Bioorg Med
Chem Lett 22:110–114
123