Design, synthesis and biological activities of thiourea containing sorafenib analogs as antitumor agents

Article abstract of DOI:10.1016/j.bmc.2012.03.018

A novel series of diaryl thiourea containing sorafenib derivatives 9a-t was designed and synthesized. The structures of all the newly synthesized compounds were determined by ¹H NMR, ¹³C NMR and HRMS. Their antiproliferative activities against HCT116 and MDA-MB-231 cell lines, and their inhibitory activities against the phosphorylation of VEGFR were evaluated and described. Some of the compounds showed significant activities against both cell lines and VEGFR. Compounds 9g, 9m, 9o and 9p demonstrated competitive antiproliferative activities to sorafenib, the reference standard, while compounds 9d, 9m, and 9p showed significant inhibitory activities against the phosphorylation of VEGFR.

Full text of DOI:10.1016/j.bmc.2012.03.018

Bioorganic & Medicinal Chemistry 20 (2012) 2923–2929
Contents lists available at SciVerse ScienceDirect
Bioorganic & Medicinal Chemistry
journal homepage: www.elsevier.com/locate/bmc
Design, synthesis and biological activities of thiourea containing sorafenib
analogs as antitumor agents
Jianwen Yao a,b, Jing Chen b, Zuopeng He b, Wei Sun a, Wenfang Xu a,
⇑
a Department of Medicinal Chemistry, School of Pharmaceutical Sciences, Shandong University, 44 West Culture Road, Jinan, Shandong 250012, PR China
b Department of Medicinal Chemistry, School of Pharmacy, Yantai University, 32 Qingquan Road, Yantai, Shandong 264005, PR China
a r t i c l e i n f o
a b s t r a c t
Article history:
A novel series of diaryl thiourea containing sorafenib derivatives 9a–t was designed and synthesized. The
structures of all the newly synthesized compounds were determined by 1H NMR, 13C NMR and HRMS.
Their antiproliferative activities against HCT116 and MDA-MB-231 cell lines, and their inhibitory activ-
ities against the phosphorylation of VEGFR were evaluated and described. Some of the compounds
showed significant activities against both cell lines and VEGFR. Compounds 9g, 9m, 9o and 9p demon-
strated competitive antiproliferative activities to sorafenib, the reference standard, while compounds
9d, 9m, and 9p showed significant inhibitory activities against the phosphorylation of VEGFR.
Ó 2012 Elsevier Ltd. All rights reserved.
Received 20 January 2012
Revised 6 March 2012
Accepted 6 March 2012
Available online 14 March 2012
Keywords:
Sorafenib analogs
Synthesis
Diaryl thiourea
Antitumor agents
1. Introduction
2. Chemistry
Due to the diverse side effects of traditional cytotoxic antican-
cer drugs, it is necessary to exploit novel antitumor drugs with
high efficiency and low toxicity. Recent advances and applications
on multiple targeted agents may afford effective chemotherapeu-
tics with low toxicity.
The synthetic routes to the library compounds were illustrated
as outlined in Scheme 1.
The substituted diaromatic ethers were prepared in a four-step se-
quence. The starting material 2-picolinic acid 1 was treated with SOCl2
to generate 4-chloropicolinoyl chloride 2, and then treated with meth-
anol to produce methyl 4-chloropicolinate 3. Subsequently, 3 was trea-
ted with corresponding amine (methylamine, n-butylamine,
phenylmethanamine, cyclopropanamine and cyclohexanamine) to
form 4a–e. Then, they were treated with 4-aminophenol to generate
corresponding diaromatic ethers (5a–e) with the total yields of 32–
54%. Furthermore, substituted anilines(2,4-dichloroaniline, 4-chloro-
aniline, 3,4-difluoroaniline, 4-fluoroaniline, 3-(trifluoromethyl)aniline,
3,5-bis(trifluoromethyl)aniline, 4-chloro-3-(trifluoromethyl)aniline
and 4-methoxyaniline), 6a–h, were treated with CS2 to generate 7a–
h, and then treated with triphosgene (BTC) to produce various isothio-
cyanates (8a–h). Finally, these isothiocyanates were reacted with
substituted diaromatic ethers (5a–e) in DCM to afford thiourea (9a–
t) in 19–48% total yields. The final products were purified by column
chromatography and their structures were characterized by 1H NMR,
13C NMR and HRMS.
Sorafenib, a diaryl urea multiple-targeted antitumor agent, can
inhibit several kinases involved in tumor proliferation and angio-
genesis including Raf, VEGFR, PDGFR and KIT.1–4 Due to the advan-
tages of multi-mechanisms, broad-spectrum anticancer potency,
and well-tolerated results in combination trials, more and more
researchers have focused on the optimization of sorafenib.5–10
Inspired by the classic bioisosteric paradigm of thiopental
replacing urea in pentobarbital with thiourea, a series of novel
sorafenib derivatives were designed and synthesized by substitut-
ing thiourea for urea in sorafenib. The antitumor effect of all the
newly synthesized compounds on the in vitro growth of two cell
lines, namely human colorectal carcinoma (HCT116) and human
breast cancer cell (MDA-MB-231) was evaluated. Apparent growth
inhibition was observed for most of the compounds, with 9d, 9g,
9m, 9o and 9p demonstrating more potent activities against cancer
cells as compared to sorafenib, respectively. Furthermore, some
structure–activity relationships have also been established.
3. Results and discussion
3.1. Cell inhibition
⇑
In vitro cell cytotoxicity of the 20 new diaryl thiourea deriva-
tives was initially evaluated against HCT116 and MDA-MB-231
Corresponding author. Tel./fax: +86 531 88382264.
E-mail address: wfxu@yahoo.cn (W. Xu).
0968-0896/$ - see front matter Ó 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.bmc.2012.03.018

2924
J. Yao et al. / Bioorg. Med. Chem. 20 (2012) 2923–2929
Cl
Cl
b
a
N
1
COOH
N
2
COCl
N
3
COOCH3
Cl
N
O
O
R1
c
d
N
H
H
N
N
R1
H2N
SH
R2
H
N
H
N
O
g
N
4a-4e
5a-5e
S
NH R1
R2
O
R2
O
R2
S
C
e
f
H
N
9a-9t
NCS
NH2
6a-6h
8a-8h
7a-7h
Scheme 1. Reagents and conditions: (a) SOCl2 (b) CH3OH (c) RNH2, THF (d) 4-aminophenol, DMF, KOBu-t (e) Dabco, CS2, toluene (f) BTC, DCM (g) DCM.
cells by MTT assay using sorafenib as a positive control. As shown
in Table 1, most compounds (except 9b, 9e, 9l and 9t) exhibited po-
tent antiproliferative activity against HCT116 cells with
9d and 9p can inhibit the phosphorylation of VEGFR at the concen-
tration of 1 g/mL. All these three compounds (9d, 9m, and 9p)
l
have the methyl substitute on the terminal amide, and their substi-
tutes in phenyl ring are 4-Cl (9d), 3-CF3 (9m) or both of them (9p).
These results indicate that the methyl on the terminal amide, 4-Cl
and 3-CF3 on the phenyl ring contribute more to the phosphoryla-
tion inhibition of VEGFR. And these compounds also showed po-
tent cytotoxicity against HCT116 and MDA-MB-231 cells as
mentioned above. While other compounds such as 9g and 9o
which have no phosphorylation inhibition of VEGFR also have sig-
nificant cytotoxicities, which indicates that the compounds 9g and
9o have the other mechanism to inhibit the growth of cancer cells,
which need further investigation.
IC50 = 9.15–53.2 lM. Compounds 9d, 9g, 9m, 9o and 9p showed
significant inhibitory activities against both cell lines. Among
them, two compounds 9o and 9p demonstrated competitive anti-
proliferative activities to sorafenib, the reference standard. The ba-
sic structure–activity relationships (SAR) were also studied: (1)
The size and shape of the hydrophobic R group: a. The longer sub-
stitutes (such as n-butyl and benzyl groups) of R might cause a de-
crease in cytotoxicity in general. For example, among the 20
compounds, only 9b, 9e, 9l and 9t whose R substitutes are n-butyl
and benzyl groups have no antiproliferative activity to HCT116 and
MDA-MB-231 cells. b. The size and shape of R group may influence
the selectivity on different cancer cells. It is notable that only the
compounds having methyl group show the inhibitory activity
against MDA-MB-231 cell, and these compounds also have potent
4. Conclusions
In summary, we report here the synthesis and biological evalu-
ation of a new class of sorafenib derivatives as potential antitumor
agents. Most compounds exhibited good antiproliferative activity
to HCT116 cells. Compounds 9d, 9g, 9m, 9o and 9p which contains
methyl group on the terminal amide showed significant inhibitory
activities against both HCT116 and MDA-MB-231 cell lines, and
two compounds 9o and 9p demonstrated competitive antiprolifer-
ative activities to sorafenib. Notably, compared with sorafenib,
most of the other diaryl thiourea compounds showed excellent
selectivity toward HCT-116 over MDA-MB-231 cells. On the other
hand, 9d, 9m, and 9p showed inhibition against the phosphoryla-
tion of VEGFR. These three compounds also have remarkable anti-
proliferative activities against both HCT116 and MDA-MB-231 cell
lines, while compounds 9g and 9o which also have good cytotoxic-
ities have no phosphorylation inhibition of VEGFR, which indicates
that the compounds 9g and 9o have the other mechanism to inhi-
bit the growth of cancer cells. It is of interest to continue studying
cytotoxicity and kinase activities of diaryl thiourea derivatives.
activities against HCT-116 cell line (IC50 = 16.1–30.6 lM). While
the other compounds (except for 9b, 9e, 9l and 9t) can only inhibit
the growth of HCT116 cells. This trait is especially obvious in the
compounds containing cyclohexyl ring. The compounds 9j, 9n
and 9r containing cyclohexyl group showed good antiproliferative
activity to HCT116 (IC50 = 9.15–26.15 lM), which are even better
than those of compounds containing methyl group. (2) The substi-
tutes on the terminal phenyl ring: a. Apart from the terminal R
group, the terminal Ar group can also affect the selectivity against
cancer cells. The compounds against MDA-MB-231 cells not only
contained methyl group of R terminal, but also contained 3,4-diflu-
o-substitue, 3-CF3, 4-Cl or both of them on the phenyl ring. b. The
substitutes on the terminal phenyl ring may also influence the
activity against HCT116 cells. For example, the compound 9s con-
taining the hydrophobic group (-OCH3) on the 4-position of phenyl
ring has less cytotoxicity compared with other compounds (9a, 9d,
9g, 9k, 9m, 9o and 9p) which containing the same R group with 9s.
The influence of substitutes on phenyl ring still needs further
investigation.
5. Experimental
5.1. Cell inhibition assays
3.2. Phosphorylation inhibition of VEGFR
HCT116 and MDA-MB-231 cell lines were plated on 96-well
plates at a density of 5000 per well and incubated overnight. The
cells were treated with compounds and sorafenib at final concen-
trations ranging from 0.5 to 200 lM, while control cells were trea-
ted with equal volume DMSO. After 48 hr treatment, 0.5% MTT
The inhibition against the phosphorylation of VEGFR was tested
in cell level using western blotting analysis. Compounds 9d, 9m,
and 9p showed inhibition against the phosphorylation of VEGFR.
As shown in Figure 1, sorafenib can inhibit the phosphorylation
of VEGFR at the concentration of 0.01
lg/mL; 9m can partly inhibit
(Amresco, USA) solution was added to each well, and further
the phosphorylation of VEGFR at the concentration of 0.1
l
g/mL;

J. Yao et al. / Bioorg. Med. Chem. 20 (2012) 2923–2929
2925
Table 1
The structures and IC50 values of the target compounds
H
N
H
N
Ar
N
S
NH R
O
O
Compd No.
Substituent
R
IC50
(
l
M)a
Compd No.
Substituent
IC50 (l
M)a
Ar
Cl
HCT116
MDA-MB-231
Ar
R
HCT116
>200
MDA-MB-231
9a
9b
9c
21.7 1.2
>200
>200
>200
9l
>200
Me
Bu
Cl
Cl
Cl
Cl
F
Cl
Cl
CF3
9m
22.6 0.9
64.9 1.8
Me
CF3
CF3
19.6 2.1
30.6 0.9
>200
53.2 1.0
17.3 1.8
>200
9n
9o
122.5 7.8
33.4 1.5
Me
Bu
Me
Me
9d
CF3
CF3
9e
9f
Cl
Cl
>200
>200
9p
25.3 1.0
29.7 0.7
Cl
CF3
24.6 0.8
16.1 2.5
32.5 1.4
>200
15.9 1.5
9.15 0.7
43.1 1.9
>200
>200
>200
9q
9r
Cl
CF3
F
9g
9h
80.1 4.2
>200
Me
Bu
F
F
Cl
F
9s
Me
OCH3
OCH3
F
F
9i
9j
52.5 2.0
>200
9t
>200
>200
F
Sorafenib
—
—
7.8 0.8
36 2.1
26.15 3.1
31.1 1.1
>200
>200
F
F
Me
9k
a
Mean value of three experiments and standard deviation are given.
+
sorafenib 9d
9m
9p
VEGFR
VEGFR
VEGFR
5.2. Phosphorylation inhibition of VEGFR
0.01 µg/mL
0.1 µg/mL
HUVEC cells were incubated with different concentrations of
compounds for 2 h. Cells in each group were harvested, washed
twice with ice-cold PBS, and lysed with cell lysis buffer except
for the blank group after 5 min of addition of VEGF. Then western
blotting was performed to test the VEGFR using anti-phospho-
VEGF receptor 2 as the antibody.
1 µg/mL
5.3. Chemistry: general procedures
Figure 1. Western blots showing protein expression of VEGFR after treatment with
compounds 9d, 9m, 9p and sorafenib at various concentrations (0.01, 0.1 and 1 lg/
mL).
All the material we used were purchased from commercial sup-
pliers and used without further purification. Solvents were distilled
prior to use and flash chromatography was performed using silica
gel (60 Å, 200–300 mesh). All reactions were monitored by thin-
layer chromatography on 0.25 mm silica gel plates (60GF-254)
and visualized with UV light, or chloride ferric. Melting points were
incubation for 4 h, then cells centrifuged at 2,500 rpm for 15 min,
removed the culture medium, and added 150 L DMSO to dissolve
the formazan. After mixing for 5 min, optical density was detected
l
at 570 nm on a microplate reader (Thermo, USA).

2926
J. Yao et al. / Bioorg. Med. Chem. 20 (2012) 2923–2929
determined on an electrothermal melting point apparatus and
were uncorrected. Proton nuclear magnetic resonance (1H NMR)
spectra were determined on a Brucker Avance 600 spectrometer
using TMS as an internal standard in DMSO-d6 solutions. Chemical
shifts are reported in delta (d) units, parts per million (ppm) down-
field from trimethylsilane. ESI-MS were determined on an API 4000
spectrometer. High-resolution mass spectral (HRMS) data were
conducted by Shandong Analysis and Test Center, and are reported
as m/z (relative intensity).
5.3.7. 4-(4-Aminophenoxy)-N-methylpicolinamide (5a)
A solution of 4-aminophenol (10.8 g, 99 mmol) in dry N,N-
dimethylformamide (DMF, 100 mL) was treated with potassium
tert-butoxide (11.2 g, 100 mmol), and the mixture was stirred at
room temperature for 1 h under nitrogen atmosphere, then a solu-
tion of (4a) (16.0 g, 94 mmol) in DMF (50 mL) and potassium
carbonate were added respectively. The mixture was heated to
80–85 °C for 10 h. After the temperature was cooled to room tem-
perature, the mixture was diluted with water (400 mL) and ex-
tracted with EtOAc (3 ꢀ 200 mL). The extract was washed with
brine (2 ꢀ 200 mL), dried over anhydrous sodium sulfate, and con-
centrated under reduced pressure to give light-brown solid (5a)
(20.3 g, 88.6%); mp: 109–112 °C. 1H NMR (400 MHz, DMSO-d6) d
3.01 (d, J = 4.8 Hz, 3H), 3.74 (br d, 2H), 6.68, 6.87 (AA0BB0, q,
J = 8.7 Hz, 4H), 7.01 (dd, J = 5.0, 2.2 Hz, 1H), 7.63 (d, J = 2.2 Hz,
1H), 8.44 (d, J = 5.0 Hz, 1H), 8.75 (br d, J = 4.6 Hz, 1H).
5.3.1. Methyl 4-chloropicolinate (3)
2-Picolinic acid (15 g, 0.122 mol) was slowly added to thionyl
chloride (50 mL) at a temperature range of 45–50 °C under stirring.
Then the solution was heated to 72 °C, and kept refluxing for about
14 h. After cooled to room temperature, the mixture was diluted
with toluene (100 mL), and concentrated to near dryness in vacuo.
To the obtained oily residue, methanol (40 mL) was added. The
contents were stirred for 2 h at 30 °C. After filtration, the filter cake
was added 15% Na2CO3 solution to adjust its pH value to neutral,
filtrate and washed with cold methanol to obtained light yellow
solid 17.5 g (83.7%), mp: 52–54 °C. 1H NMR (400 MHz, DMSO-d6)
d 3.88 (s, 3H); 7.89 (dd, J = 5.4, 2.0 Hz, 1H); 8.10 (d, J = 2.1 Hz,
1H); 8.70 (d, J = 5.4 Hz, 1H).
5.3.8. 4-(4-Aminophenoxy)-N-butylpicolinamide (5b)
The synthesis is analogous to (5a) with (4b) as the starting
material. The product was obtained as a brown solid. Yield = 73.5%.
Mp: 85–88 °C. 1H NMR (400 MHz, DMSO-d6) d 0.94 (t, J = 7.3 Hz,
3H), 1.39 (m, 2H), 1.58 (m, 2H), 3.42 (q, J = 6.8 Hz, 2H), 3.72 (br
d, 2H), 6.69, 6.87 (AA0BB0, q, J = J = 8.7 Hz, 4H), 6.92 (dd, J = 5.6,
2.4 Hz, 1H), 7.67 (d, J = 2.4 Hz, 1H), 8.03 (br d, 1H), 8.32 (d,
J = 5.6 Hz, 1H).
5.3.2. 4-Chloro-N-methylpicolinamide (4a)11
The compound 3 (10 g, 58.3 mmol) was dissolved in 70 mL
methanol/THF (2/5) mixture at 0 °C, then a solution of methyl-
amine (5 g, 161.3 mmol) in THF (60 mL) was added at a rate that
kept the internal temperature below 2 °C. The mixture was stirred
at room temperature for 3 h and then was concentrated in vacuo.
The crude product was dissolved in 100 mL ethyl acetate and
was washed with brine, dried over anhydrous sodium sulfate,
and concentrated under reduced pressure to afford 4a (9.4 g,
94.5%) as light yellow crystalline. Mp: 41–43 °C. 1H NMR
(400 MHz, DMSO-d6) d 2.80 (d, J = 4.6 Hz, 3H), 7.78 (dd, J = 5.2,
2.1 Hz, 1H); 8.05 (d, J = 2.1 Hz, 1H), 8.62 (d, J = 5.2 Hz, 1H), 8.85
(br d, J = 4.6 Hz, 1H); ESI-MS m/z 171([M+H]+).
5.3.9. 4-(4-Aminophenoxy)-N-benzylpicolinamide (5c)
The synthesis is analogous to (5a) with (4c) as the starting
material. The product was obtained as a yellow solid. Yield = 67.7%.
Mp: 71–74 °C. 1H NMR (400 MHz, DMSO-d6) d 3.72 (br d, 2H), 4.63
(d, J = 6.0 Hz, 2H), 6.70, 6.87 (AA0BB0, q, J = 8.7 Hz, 4H), 6.92 (dd,
J = 5.6, 2.6 Hz, 1H), 7.26 (m, 1H), 7.33 (m, 4H), 7.70 (d, J = 2.6 Hz,
1H), 8.32 (d, J = 5.6 Hz, 1H), 8.38 (t, J = 6.2 Hz, 1H). ESI-MS m/z
320.1 [M+H]+.
5.3.10. 4-(4-Aminophenoxy)-N-cyclopropylpicolinamide (5d)
The synthesis is analogous to (5a) with (4d) as the starting
material. The product was obtained as a yellow solid. Yield = 72.5%.
Mp: 92–95 °C. 1H NMR (400 MHz, DMSO-d6) d 0.83 (d, J = 7.2 Hz,
4H), 2.81 (m, 1H), 3.73 (br d, 2H), 6.67, 6.86 (AA0BB0, q, J = 8.7 Hz,
4H), 7.17 (dd, J = 5.6, 2.4 Hz, 1H), 7.40 (d, J = 2.4 Hz, 1H), 8.58 (d,
J = 5.6 Hz, 1H), 8.68 (br d, 1H). ESI-MS m/z 270.1 [M+H]+.
5.3.3. N-Butyl-4-chloropicolinamide (4b)
The synthesis is analogous to (4a) with n-butylamine as the
starting material. The product was obtained as a orange solid.
Yield = 82.9%. Mp: 42 – 45 °C. 1H NMR (400 MHz, DMSO-d6) d
0.97 (t, J = 7.3 Hz, 3H), 1.40 (m, 2H), 1.59 (m, 2H), 3.42 (q,
J = 6.8 Hz, 2H), 7.39 (dd, J = 5.5, 2.2 Hz, 1H), 7.72 (d, J = 2.2 Hz,
1H), 8.05 (br d, 1H), 8.40 (d, J = 5.5 Hz, 1H).
5.3.11. 4-(4-Aminophenoxy)-N-cyclohexylpicolinamide (5e)
The synthesis is analogous to (5a) with (4e) as the starting
material. The product was obtained as a yellow solid. Yield = 64.9%.
Mp: 135–138 °C. 1H NMR (400 MHz, DMSO-d6) d 1.25–1.48 (m,
6H), 1.78 (m, 4H), 3.72 (br d, 2H), 3.80 (m, 1H), 6.77 (q,
J = 8.7 Hz, 4H), 7.24 (dd, J = 5.5, 2.4 Hz, 1H), 7.54 (d, J = 2.4 Hz,
1H), 8.54 (d, J = 5.5 Hz, 1H), 8.76 (br d, 1H). ESI-MS m/z 312.1
[M+H]+.
5.3.4. N-Benzyl-4-chloropicolinamide (4c)
The synthesis is analogous to (4a) with phenylmethanamine as the
starting material. The product was obtained as a light yellow solid.
Yield = 70.8%. Mp: 78–82 °C. 1H NMR (400 MHz, DMSO-d6) d 4.62 (d,
J = 6.0 Hz, 2H), 7.09 (dd, J = 5.5, 2.5 Hz, 1H), 7.25 (m, 1H), 7.32 (m, 4H),
7.73 (d, J = 2.5 Hz, 1H), 8.35 (d, J = 5.5 Hz, 1H), 8.38 (t, J = 6.2 Hz, 1H).
5.3.12. 2,4-Dichloro-1-isothiocyanatobenzene (8a)
5.3.5. 4-Chloro-N-cyclopropylpicolinamide (4d)
To a solution of 2,4-dichloroaniline (2.51 g, 15.5 mmol) and
triethylenediamine (2.08 g, 18.6 mmol) in 40 mL toluene, CS2
(3.5 g, 46.5 mmol) was added slowly. The resulting mixture was
stirred at room temperature for 8 h. After filtration, the filter cake
was washed with toluene and dried. Obtained 7a was suspended in
40 mL DCM. Then, a 25 mL DCM solution of BTC (5.0 g, 17.0 mmol)
was added slowly under ꢁ5–0 °C. The mixture was stirred for 2 h
at room temperature, and then was refluxed for 1.5–2 h. After fil-
tration, the filtrate was concentrated in vacuo. The crude product
was purified by chromatography (ethyl acetate/petroleum
ether = 1:1) on silica gel to give 0.19 g white solid (60%). Mp: 37–
42 °C.
The synthesis is analogous to (4a) with cyclopropanamine as
the starting material. The product was obtained as a yellow solid.
Yield = 90.1%. Mp: 99–102 °C. 1H NMR (400 MHz, DMSO-d6) d
0.86 (d, J = 7.0 Hz, 4H), 2.88 (m, 1H), 7.21 (dd, J = 5.5, 2.4 Hz, 1H),
8.04 (d, J = 2.4 Hz, 1H), 8.65 (d, J = 5.6 Hz, 1H), 8.80 (br d, 1H).
5.3.6. 4-Chloro-N-cyclohexylpicolinamide (4e)
The synthesis is analogous to (4a) with cyclohexanamine as the
starting material. The product was obtained as a yellow solid.
Yield = 84.6%. Mp: 117–119 °C. 1H NMR (400 MHz, DMSO-d6) d 1.26–
1.48 (m, 6H), 1.85 (m, 4H), 3.84 (m, 1H), 7.64 (dd, J = 5.4, 2.3 Hz, 1H),
8.06 (d, J = 2.3 Hz, 1H), 8.58 (d, J = 5.4 Hz, 1H), 8.79 (br d, 1H).

J. Yao et al. / Bioorg. Med. Chem. 20 (2012) 2923–2929
2927
5.3.13. 1-Chloro-4-isothiocyanatobenzene (8b)
The synthesis is analogous to (8a) with 4-chloroaniline as the
starting material. The product was obtained as a light brown solid.
Yield = 84.7%; mp: 43–45 °C.
8.80 (t, J = 6.0 Hz, 1H), 9.60 (s, 1H), 10.14 (s, 1H); 13C NMR
(300 MHz, DMSO-d6) d 14.62, 20.47, 32.11, 39.40, 109.98, 115.22,
122.01, 126.88, 128.34, 129.90, 131.96, 132.34, 136.63, 137.68,
150.91, 151.42, 153.41, 164.64, 166.60, 181.51; HRMS(AP-ESI)
m/z: calcd for C23H23Cl2N4O2S [M+H]+ 489.0914, found 489.0916.
HPLC purity = 99.04%.
5.3.14. 1,2-Difluoro-4-isothiocyanatobenzene (8c)
The synthesis is analogous to (8a) with 3,4-difluoroaniline as
the starting material. The product was obtained as a light yellow
liquid. Yield = 74.5%; bp: 170 °C.
5.3.22. N-Benzyl-4-(4-(3-(2,4-dichlorophenyl)thioureido)
phenoxy)picolinamide (9c)
Light yellow solid (yield 75.0%), mp: = 116–118 °C; 1H NMR
(600 MHz, DMSO-d6) d 4.46 (d, J = 6.2 Hz, 2H), 7.20 (dd, J = 5.6,
2.5 Hz, 1H), 7.21 (m, 1H), 7.23 (d, J = 8.8 Hz, 2H), 7.29 (m, 4H),
7.43 (d, J = 2.5 Hz, 1H), 7.45 (dd, J = 7.2, 2.3 Hz ,1H), 7.59 (d,
J = 7.2 Hz, 1H), 7.62 (d, J = 8.6 Hz, 2H), 7.71 (d, J = 2.3 Hz, 1H),
8.55 (d, J = 5.6 Hz, 1H), 9.38 (t, J = 6.4 Hz, 1H), 9.59 (s, 1H), 10.13
(s, 1H); 13C NMR (300 MHz, DMSO-d6) d 43.39, 110.08, 115.42,
122.04, 126.88, 127.72, 128.30, 128.36, 129.19, 129.90, 131.97,
132.36, 136.64, 137.70, 140.35, 150.88, 151.47, 153.32, 164.30,
5.3.15. 1-Fluoro-4-isothiocyanatobenzene (8d)
The synthesis is analogous to (8a) with 4-fluoroaniline as the
starting material. The product was obtained as a light yellow solid.
Yield = 88.5%; mp: 25–28 °C.
5.3.16. 1-Isothiocyanato-3-(trifluoromethyl)benzene (8e)
The synthesis is analogous to (8a) with 3-(trifluoromethyl)ani-
line as the starting material. The product was obtained as a light
yellow liquid. Yield = 74.3%; bp: 206–208 °C.
166.64, 181.46; HRMS(AP-ESI) m/z: calcd for
C26H21Cl2N4O2S
[M+H]+ 523.0757, found 523.0758. HPLC purity = 98.7%.
5.3.17. 1-Isothiocyanato-3,5-bis(trifluoromethyl)benzene (8f)
The synthesis is analogous to (8a) with 3,5-bis(trifluoro-
methyl)aniline as the starting material. The product was obtained
as a colorless liquid. Yield = 68.0%; bp 63 °C (2.5 mmHg).
5.3.23. 4-(4-(3-(4-Chlorophenyl)thioureido)phenoxy)
-N-methylpicolinamide (9d)
Light yellow solid (yield 69.8%), mp: = 146–148 °C; 1H NMR
(600 MHz, DMSO-d6) d 2.79 (d, J = 4.8 Hz, 3H), 7.19 (dd, J = 5.6,
2.5 Hz, 1H), 7.24 (d, J = 8.8 Hz ,2H), 7.39 (d, J = 8.6 Hz ,2H), 7.43
(d, J = 2.5 Hz, 1H), 7.52 (d, J = 8.6 Hz, 2H), 7.60 (d, J = 8.8 Hz, 2H),
8.53 (d, J = 5.6 Hz, 1H), 8.81 (br d, J = 4.8 Hz, 1H), 9.98 (s, 2H); 13C
5.3.18. 1-Chloro-4-isothiocyanato-2-(trifluoromethyl)benzene
(8g)
The synthesis is analogous to (8a) with 4-chloro-3-(trifluoro-
methyl)aniline as the starting material. The product was obtained
as a light yellow liquid. Yield = 81.0%; bp 248–251 °C.
NMR (300 MHz, DMSO-d6)
d 26.96, 109.90, 115.28, 121.96,
126.28, 126.66, 129.31, 137.96, 139.37, 150.77, 151.29, 153.54,
164.11, 166.64, 180.79; HRMS(AP-ESI) m/z: calcd for
5.3.19. 1-Isothiocyanato-4-methoxybenzene (8h)
The synthesis is analogous to (8a) with 4-methoxyaniline as the
starting material. The product was obtained as a colorless oily li-
quid. Yield = 78.6%; bp: 280 °C.
C
20H18ClN4O2S [M+H]+ 413.0834, found 413.0840. HPLC
purity = 97.8%.
5.3.24. N-Butyl-4-(4-(3-(4-
chlorophenyl)thioureido)phenoxy)picolinamide (9e)
5.3.20. 4-(4-(3-(2,4-Dichlorophenyl)thioureido)phenoxy)-N-
methylpicolinamide (9a)
Light yellow solid (yield 67.6%), mp: = 110–112 °C; 1H NMR
(600 MHz, DMSO-d6) d 0.88 (t, J = 7.2 Hz, 3H), 1.28 (m, 2H), 1.49
(m, 2H), 3.26 (q, J = 6.6 Hz, 2H), 7.18–7.22 (m, 3H), 7.38–7.42 (m,
3H), 7.52 (d, J = 8.4 Hz, 2H), 7.59 (d, J = 8.4 Hz, 2H), 8.53 (d,
J = 5.4 Hz, 1H), 8.79 (t, J = 6.6 Hz, 1H), 9.97 (s, 2H); 13C NMR
(300 MHz, DMSO-d6) d 14.60, 20.50, 32.16, 39.44, 109.87, 115.25,
121.93, 126.27, 126.68, 129.28, 137.95, 139.38, 150.72, 151.33,
153.51, 164.06, 166.63, 180.78; HRMS(AP-ESI) m/z: calcd for
To a 10 mL DCM solution of compound (5a) (1.5 g, 6.16 mmol),
a DCM solution of (8a) (1.26 g, 6.16 mmol) was added slowly at 0–
5 °C under nitrogen atmosphere. The mixture was stirred for 2 h in
ice bath, then stirred for another 18 h at room temperature. The
mixture was poured into petroleum ether (60 mL) and then fil-
tered. The solids were washed with DCM (2 ꢀ 5 mL), and dried un-
der vacuum for 6 h at 50 °C to afford light brown solid in the yield
of 69.4%. Mp: 165–167 °C. 1H NMR (600 MHz, DMSO-d6) d 2.79 (d,
J = 4.8 Hz, 3H), 7.19 (dd, J = 5.6, 2.5 Hz, 1H), 7.25 (d, J = 8.8 Hz ,2H),
7.43 (d, J = 2.5 Hz, 1H), 7.45 (dd, J = 8.6, 2.3 Hz ,1H), 7.59 (d,
J = 8.4 Hz, 1H), 7.61 (d, J = 8.7 Hz, 2H), 7.71 (d, J = 2.3 Hz, 1H),
8.53 (d, J = 5.6 Hz, 1H), 8.81 (br d, J = 4.8 Hz, 1H), 9.59 (s, 1H),
10.13 (s, 1H); 13C NMR (300 MHz, DMSO-d6) d 26.95, 109.91,
115.18, 121.97, 126.87, 128.34, 129.89, 131.95, 132.31, 136.64,
137.69, 150.95, 151.39, 153.49, 164.71, 166.56, 181.50;
HRMS(AP-ESI) m/z: calcd for C20H17Cl2N4O2S [M+H]+ 447.0444,
found 447.04547. HPLC purity = 98.34%.
C
23H24ClN4O2S [M+H]+ 455.1303, found 455.1298; HPLC
purity = 98.3%.
5.3.25. N-Benzyl-4-(4-(3-(4-chlorophenyl)thioureido)
phenoxy)picolinamide (9f)
Light yellow solid (yield 66.3%), mp: = 164–166 °C; 1H NMR
(600 MHz, DMSO-d6) d 4.46 (d, J = 6.2 Hz, 2H), 7.20 (dd, J = 5.6,
2.3 Hz, 1H), 7.21 (m, 1H), 7.23 (d, J = 8.6 Hz, 2H), 7.29 (m, 4H),
7.39 (d, J = 8.6 Hz ,2H), 7.43 (d, J = 2.3 Hz, 1H), 7.52 (d, J = 8.6 Hz,
2H), 7.58 (d, J = 8.6 Hz, 2H), 8.55 (d, J = 5.6 Hz, 1H), 9.39 (t,
J = 6.3 Hz, 1H), 9.98 (s, 2H); 13C NMR (300 MHz, DMSO-d6) d
43.40, 110.02, 115.43, 121.96, 126.28, 126.69, 127.72, 128.30,
129.19, 129.29, 137.97, 139.37, 140.35, 150.66, 151.46, 153.31,
164.31, 166.67, 180.76; HRMS(AP-ESI) m/z: calcd for
The other compounds of (9) series were synthesized following
the general procedure as described above.
5.3.21. N-Butyl-4-(4-(3-(2,4-dichlorophenyl)thioureido)
phenoxy)picolinamide (9b)
C
26H22ClN4O2S [M+H]+ 489.1147, found 489.1142. HPLC
purity = 99.21%.
Light yellow solid (yield 67.2%), mp: = 161–164 °C; 1H NMR
(600 MHz, DMSO-d6) d 0.88 (t, J = 7.3 Hz, 3H), 1.28 (m, J = 7.4 Hz,
2H), 1.49 (m, J = 7.3 Hz, 2H), 3.26 (q, J = 6.7 Hz, 2H), 7.19 (dd,
J = 5.6, 2.5 Hz, 1H), 7.23 (d, J = 8.8 Hz ,2H), 7.42 (d, J = 2.5 Hz,
1H),7.45 (dd, J = 8.6, 2.3 Hz ,1H), 7.59 (d, J = 8.4 Hz, 1H), 7.62 (d,
J = 8.6 Hz, 2H), 7.71 (d, J = 2.3 Hz, 1H), 8.53 (d, J = 5.6 Hz, 1H),
5.3.26. 4-(4-(3-(3,4-Difluorophenyl)thioureido)phenoxy)-N-
methylpicolinamide (9g)
Light yellow solid (yield 83.8%), mp: = 144–146 °C; 1H NMR
(600 MHz, DMSO-d6) d 2.79 (d, J = 4.8 Hz, 3H), 7.19 (dd, J = 5.6,
2.6 Hz, 1H), 7.20 (d, J = 2.1 Hz, 1H), 7.24 (d, J = 8.8 Hz, 2H), 7.38(d,

2928
J. Yao et al. / Bioorg. Med. Chem. 20 (2012) 2923–2929
J = 8.5 Hz 1H), 7.43 (d, J = 2.5 Hz, 1H), 7.58 (d, J = 8.8 Hz, 2H), 7.70
(m, 1H), 8.53 (d, J = 5.6 Hz, 1H), 8.79 (J = 4.8 Hz 1H), 9.99 (s, 1H),
10.01 (s, 1H); 13C NMR (300 MHz, DMSO-d6) d 26.96, 109.95,
114.07, 114.30, 115.35, 117.81, 118.04, 121.48 (q, J = 12), 121.97,
126.87, 137.38 (dd, J = 33, 12), 137.76, 150.85, 151.11, 153.47,
163.06, 165.99, 180.97; HRMS(AP-ESI) m/z: calcd for
5.3.31. N-Benzyl-4-(4-(3-(4-fluorophenyl)thioureido)
phenoxy)picolinamide (9l)
Light-brown solid (yield 87.4%), mp: = 104–107 °C; 1H NMR
(600 MHz, DMSO-d6) d 4.46 (d, J = 6.3 Hz, 2H), 7.18 (dd, J = 5.6,
2.5 Hz, 1H), 7.19 (m, 1H), 7.21 (d, J = 8.8 Hz, 2H), 7.23 (d,
J = 8.8 Hz ,2H), 7.30 (m, 4H), 7.43 (d, J = 2.5 Hz, 1H), 7.48 (d,
J = 8.8 Hz, 2H), 7.58 (d, J = 8.8 Hz, 2H), 8.55 (d, J = 5.6 Hz, 1H),
9.38 (t, J = 6.4 Hz, 1H), 9.86 (s, 1H), 9.89 (s, 1H); 13C NMR
(300 MHz, DMSO-d6) d 43.39, 110.10, 115.42, 115.89, 116.19,
121.95, 126.71, 127.18, 127.22 (d, J = 33), 128.30, 129.19, 136.63
(d, J = 9), 138.05, 140.35, 150.60, 151.45, 153.30, 164.31, 166.69,
C
20H17F2N4O2S [M+H]+ 415.1035, found 415.1032. HPLC
purity = 96.2%.
5.3.27. N-Butyl-4-(4-(3-(3,4-difluorophenyl)thioureido)
phenoxy)picolinamide (9h)
181.06; HRMS (AP-ESI) m/z: calcd for
C
26H22FN4O2S [M+H]+
Light yellow solid (yield 68.3%), mp = 161–163 °C; 1H NMR
(600 MHz, DMSO-d6) d 0.88 (t, J = 7.4 Hz, 3H), 1.28 (m, J = 7.5 Hz,
2H), 1.48 (m, J = 7.3 Hz, 2H), 3.26 (q, J = 6.6 Hz, 2H), 7.19 (dd,
J = 5.6, 2.5 Hz, 1H), 7.20 (d, J = 2.2 Hz, 1H), 7.23 (d, J = 8.7 Hz ,2H),
7.39 (d, J = 8.5 Hz, 1H), 7.42 (d, J = 2.5 Hz, 1H), 7.58 (d, J = 8.7 Hz,
2H), 7.70 (m, 1H), 8.53 (d, J = 5.6 Hz, 1H), 8.81 (t, J = 5.9 Hz, 1H),
9.99 (s, 1H), 10.01 (s, 1H); 13C NMR (300 MHz, DMSO-d6) d 14.61,
20.50, 32.17, 39.44, 109.94, 114.06, 114.31, 115.40, 117.80,
118.04, 121.47 (q, J = 12), 121.97, 126.88, 137.38 (dd, J = 33, 12),
137.73, 150.86, 151.11, 153.47, 163.07, 165.98, 180.98;
HRMS(AP-ESI) m/z: calcd for C23H23F2N4O2S [M+H]+ 457.1505,
found 457.1501. HPLC purity = 99.2%.
473.1442, found 473.1440. HPLC purity = 98.5%.
5.3.32. N-Methyl-4-(4-(3-(3-(trifluoromethyl)phenyl)
thioureido)phenoxy)picolinamide (9m)
White solid (yield 69.3%), mp: = 108–110 °C; 1H NMR (600 MHz,
DMSO-d6) d 2.79 (d, J = 4.8 Hz, 3H), 7.19 (dd, J = 5.6, 2.6 Hz, 1H),
7.23 (d, J = 8.8 Hz, 2H), 7.43 (d, J = 2.5 Hz, 1H), 7.48 (d, J = 7.9 Hz,
1H), 7.55 (d, J = 7.6 Hz, 1H), 7.58 (d, J = 8.8 Hz, 2H), 7.77 (m, 1H),
7.97 (s, 1H), 8.53 (d, J = 5.6 Hz, 1H), 8.81 (br d, J = 4.8 Hz 1H),
10.12 (br d, 1H), 10.13 (br d, 1H); 13C NMR (300 MHz, DMSO-d6)
d 26.96, 109.07, 115.42, 120.82, (d, J = 15), 121.62 (d, J = 15),
122.02, 126.81, 128.33, 129.74, 130.16, 130.48, 137.68, 141.35,
150.86, 151.36, 153.42, 163.02, 166.60, 180.89; HRMS (AP-ESI)
m/z: calcd for C21H18F3N4O2S [M+H]+ 447.1098, found 447.1103.
HPLC purity = 98.04%.
5.3.28. N-Cyclopropyl-4-(4-(3-(3,4-difluorophenyl)thioureido)
phenoxy)picolinamide (9i)
Light yellow solid (yield 77.9%), mp: = 162–163 °C; 1H NMR
(600 MHz, DMSO-d6) d 0.67 (d, J = 7.1 Hz, 4H), 2.86 (m, 1H), 7.19
(dd, J = 5.6, 2.6 Hz, 1H), 7.20 (d, J = 2.4 Hz ,1H), 7.23 (d, J = 8.8 Hz,
2H), 7.39 (d, J = 8.5 Hz 1H), 7.43 (d, J = 2.5 Hz, 1H), 7.58 (d,
J = 8.8 Hz, 2H), 7.70 (m, 1H), 8.51 (d, J = 5.6 Hz, 1H), 8.76 (d,
J = 5.0 Hz, 1H), 9.98 (s, 1H), 10.00 (s, 1H); 13C NMR (300 MHz,
DMSO-d6) d 6.65, 23.83, 109.96, 114.11, 114.32, 115.41, 117.80,
118.03, 121.48 (q, J = 12), 121.98, 126.89, 137.38 (dd, J = 33, 12),
137.77, 150.87, 151.08, 153.48, 163.09, 166.01, 180.98;
HRMS(AP-ESI) m/z: calcd for C22H19F2N4O2S [M+H]+ 441.1192,
found 441.1196. HPLC purity = 98.3%.
5.3.33. N-Cyclohexyl-4-(4-(3-(3-(trifluoromethyl)phenyl)
thioureido)phenoxy)picolinamide (9n)
Light yellow solid (yield 68.9%), mp: = 151–154 °C; 1H NMR
(600 MHz, DMSO-d6) d 1.11–1.16 (m, 1H) , 1.27–1.33 (m, 2H),
1.36–1.42 (m, 2H), 1.58 (d, J = 12 Hz, 1H), 1.67–1.71 (m, 2H),
1.75–1.77 (m, 2H), 3.70–3.75 (m, 1H), 7.23 (dd, J = 5.6, 2.5 Hz,
1H), 7.40 (d, J = 8.8 Hz, 2H), 7.48 (d, J = 7.9 Hz, 1H), 7.54 (d,
J = 2.5 Hz, 1H), 7.55 (d, J = 7.6 Hz, 1H), 7.68 (d, J = 8.7 Hz, 2H),
7.77 (m, 1H), 7.97 (s, 1H), 8.54 (d, J = 5.6 Hz, 1H), 8.75 (br d, 1H),,
10.12 (br d, 1H), 10.13 (br d, 1H); 13C NMR (300 MHz, DMSO-d6)
d 25.69, 26.01, 33.05, 48.92, 109.03, 115.39, 120.82, (d, J = 15),
121.62 (d, J = 15), 122.02, 126.80, 128.34, 129.74, 130.15, 130.48,
137.70, 141.37, 150.89, 151.33, 153.46, 163.06, 166.61, 180.89;
HRMS (AP-ESI) m/z: calcd for C26H26F3N4O2S [M+H]+ 515.1724,
found 515.1728. HPLC purity = 98.9%.
5.3.29. N-Cyclohexyl-4-(4-(3-(3,4-difluorophenyl)thioureido)
phenoxy)picolinamide (9j)
Light yellow solid (yield 67.0%), mp: = 162–164 °C; 1H NMR
(600 MHz, DMSO-d6) d 1.10–1.16 (m, 1H), 1.27–1.33 (m, 2H),
1.36–1.42 (m, 2H), 1.59 (d, J = 12 Hz, 1H), 1.67–1.71 (m, 2H),
1.75–1.77 (m, 2H), 3.70–3.76 (m, 1H), 7.19–7.26 (m, 4H), 7.41
(m, 2H), 7.58 (d, J = 9.0 Hz, 2H), 7.68–7.62 (m, 1H), 8.47 (d,
J = 9.0 Hz, 1H), 8.53 (d, J = 5.4 Hz, 1H), 9.98 (s, 1H), 10.00 (s, 1H);
5.3.34. 4-(4-(3-(3,5-Bis(trifluoromethyl)phenyl)
thioureido)phenoxy)-N-methylpicolinamide (9o)
White solid (yield 76.9%), mp: = 170–172 °C; 1H NMR (600 MHz,
DMSO-d6) d 2.79 (d, J = 4.8 Hz, 3H), 7.19 (dd, J = 5.6, 2.6 Hz, 1H),
7.25 (d, J = 8.8 Hz, 2H), 7.43 (d, J = 2.5 Hz, 1H), 7.58 (d, J = 8.8 Hz,
2H), 7.83 (s, 1H), 8.27 (s, 2H), 8.53 (d, J = 5.6 Hz, 1H), 8.81 (br d,
J = 4.8 Hz 1H), 10.34 (br d, 1H), 10.38 (br d, 1H); 13C NMR
(300 MHz, DMSO-d6) d 26.95, 109.87, 115.31, 122,19, 124.59,
127.10, 130.76, 131.20, 137.26, 142.78, 151.27, 151.42, 153.48,
164.71, 166.47, 180.96; HRMS (AP-ESI) m/z: calcd for
13C NMR (300 MHz, DMSO-d6)
d 25.67, 26.02, 33.05, 48.92,
109.95, 114.04, 114.31, 115.37, 117.81, 118.04, 121.46 (q, J = 12),
121.91, 126.87, 137.37 (dd, J = 36, 12), 137.79, 150.89, 151.32,
153.47, 163.07, 166.63, 180.98. HRMS (AP-ESI) m/z: calcd for
C
25H25F2N4O2S [M+H]+ 483.1661, found 483.1655. HPLC
purity = 98.9%.
C
22H17F6N4O2S [M+H]+ 515.0971, found 515.0970. HPLC
5.3.30. 4-(4-(3-(4-Fluorophenyl)thioureido)phenoxy)
-N-methylpicolinamide (9k)
purity = 99.0%.
Light yellow solid (yield 70.3%), mp: = 172–173 °C; 1H NMR
(600 MHz, DMSO-d6) d 2.79 (d, J = 4.8 Hz, 3H), 7.19 (dd, J = 5.6,
2.5 Hz, 1H), 7.21 (d, J = 8.8 Hz, 2H), 7.23 (d, J = 8.8 Hz ,2H), 7.43
(d, J = 2.5 Hz, 1H), 7.48 (d, J = 8.8 Hz, 2H), 7.59 (d, J = 8.8 Hz, 2H),
8.53 (d, J = 5.6 Hz, 1H), 8.81 (br d, J = 4.8 Hz, 1H), 9.86 (s, 1H),
9.89 (s, 1H); 13C NMR (300 MHz, DMSO-d6) d 26.96, 109.80,
115.18, 115.89, 116.19, 121.92, 126.71, 127.22 (d, J = 33), 136.63
(d, J = 12), 138.02, 150.64, 151.38, 153.45, 164.72, 166.61, 181.07;
HRMS(AP-ESI) m/z: calcd for C20H18FN4O2S [M+H]+ 397.1129,
found 397.1132. HPLC purity = 97.84%.
5.3.35. 4-(4-(3-(4-Chloro-3-(trifluoromethyl)phenyl)
thioureido)phenoxy)-N-methylpicolinamide (9p)
Light brown solid (yield 82.3%), mp: = 152–154 °C; 1H NMR
(600 MHz, DMSO-d6) d 2.79 (d, J = 4.8 Hz, 3H), 7.19 (dd, J = 5.6,
2.6 Hz, 1H),7.24 (d, J = 8.8 Hz, 2H) 7.43 (d, J = 2.5 Hz, 1H), 7.58 (d,
J = 8.8 Hz, 2H), 7.69 (d, J = 8.7 Hz, 1H), 7.82 (dd, J = 8.7, 2.1 Hz,
1H), 8.09 (d, J = 2.2 Hz, 1H), 8.53 (d, J = 5.6 Hz, 1H), 8.79 (q,
J = 4.8 Hz 1H), 10.16 (br d, 1H), 10.18 (br d, 1H); 13C NMR
(300 MHz, DMSO-d6) d 26.95, 109.68, 115.43, 122..07, 123.40 (m),

J. Yao et al. / Bioorg. Med. Chem. 20 (2012) 2923–2929
2929
125.49, 126.19, 126.96, 127.36, 129.57, 132.52, 137.55, 140.12,
151.04, 151.31, 153.33, 165.45, 166.49, 180.82; HRMS (AP-ESI)
m/z: calcd for
7.18–7.24 (m, 4H), 7.29–7.36 (m, 6H), 7.43 (d, J = 2.4 Hz, 1H), 7.57–
7.59 (m, 2H), 8.54 (d, J = 5.4 Hz, 1H), 9.36 (t, J = 6.6 Hz, 1H), 9.71 (s,
2H); 13C NMR (300 MHz, DMSO-d6) d 43.40, 56.19, 110.03, 114.68,
115.38, 121.84, 126.65, 127.01, 127.71, 128.30, 129.19, 133.02,
138.26, 140.35, 150.45, 151.44, 153.30, 157.58, 164.32, 166.72,
C
21H17ClF3N4O2S [M+H]+ 481.0708, found
481.0714. HPLC purity = 98.6%.
5.3.36. 4-(4-(3-(4-Chloro-3-(trifluoromethyl)phenyl)
180.95; HRMS(AP-ESI) m/z: calcd for C
27H25N4O3S [M+H]+
thioureido)phenoxy)-N-cyclopropylpicolinamide (9q)
Off-white solid (yield 77.6%), mp: = 149–152 °C; 1H NMR
(600 MHz, DMSO-d6) d 0.64–0.69 (m, 4H), 2.84–2.88 (m, 1H),
7.19 (dd, J = 5.8, 3.0 Hz, 1H), 7.24 (m, 2H), 7.41 (d, J = 3.0, 1H),
7.58 (m, 2H), 7.69 (d, J = 8.4 Hz, 1H), 7.82 (dd, J = 8.4, 2.4 Hz, 1H),
8.09 (d, J = 2.4, 1H), 8.51 (d, J = 5.6 Hz, 1H), 8.76 (d, J = 5.4 Hz,
1H), 10.17 (s, 1H), 10.20 (s, 1H); 13C NMR (300 MHz, DMSO-d6) d
6.66, 23.83, 109.75, 115.40, 122.10, 123.40 (m), 125.47, 126.12,
126.91, 127.37, 129.51, 132.51, 137.55, 140.13, 151.00,
151.33,153.32, 165.47, 166.52, 180.84; HRMS (AP-ESI) m/z: calcd
for C23H19ClF3N4O2S [M+H]+ 507.0864, found 507.0861. HPLC
purity = 98.5%.
485.1642, found 485.1643. HPLC purity = 97.34%.
Acknowledgments
This work was supported by National Science and Technology
Major Project (Grant No. 2009-ZX09103-118) and Ph.D. Programs
Foundation of Ministry of Education of China (No.
20110131110037).
Supplementary data
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.bmc.2012.03.018.
These data include MOL files and InChiKeys of the most important
compounds described in this article.
5.3.37. 4-(4-(3-(4-Chloro-3-(trifluoromethyl)phenyl)
thioureido)phenoxy)-N-cyclohexylpicolinamide (9r)
White solid (yield 63.8%), mp: = 178–180 °C; 1H NMR (600 MHz,
DMSO-d6) d 1.10–1.16 (m, 1H), 1.27–1.33 (m, 2H), 1.36–1.42 (m,
2H), 1.59 (d, J = 12 Hz, 1H), 1.67–1.71 (m, 2H), 1.75–1.77 (m, 2H),
3.70–3.76 (m, 1H), 7.24 (dd, J = 5.6, 2.5 Hz, 1H), 7.40 (d, J = 8.8 Hz,
2H), 7.54 (d, J = 2.5 Hz, 1H), 7.68 (d, J = 8.7 Hz, 2H), 7.70 (d,
J = 8.7 Hz, 1H), 7.82 (dd, J = 8.7, 2.1 Hz, 1H), 8.09 (d, J = 2.2 Hz,
1H), 8.54 (d, J = 5.6 Hz, 1H), 8.75 (br d, 1H), 10.17(s, 1H), 10.19(s,
1H); 13C NMR (300 MHz, DMSO-d6) d 25.68, 26.02, 33.05, 48.92,
109.76, 115.44, 122.08, 123.40 (m), 125.47, 126.12, 126.93,
127.38, 129.52, 132.54, 137.55, 140.11, 151.00, 151.30,153.34,
165.42, 166.54, 180.87; HRMS(AP-ESI) m/z: calcd for
References and notes
1. Liu, L.; Cao, Y.; Chen, C.; Zhang, X.; McNabola, A.; Wilkie, D.; Wilhelm, S.; Lynch,
M.; Carter, C. Cancer Res. 2006, 66, 11851.
2. Wilhelm, S.; Carter, C.; Lynch, M.; Lowinger, T.; Dumas, J.; Smith, R. A.;
Schwartz, B.; Simantov, R.; Kelley, S. Nat. Rev. Drug Discov. 2006, 5, 835.
3. Guida, T.; Anaganti, S.; Provitera, L.; Gedrich, R.; Sullivan, E.; Wilhelm, S. M.;
Santoro, M.; Carlomagno, F. Clin. Cancer. Res. 2007, 13, 3363.
4. Wilhelm, S. M.; Adnane, L.; Newell, P.; Villanueva, A.; Llovet, J. M.; Lynch, M.
Mol. Cancer Ther. 2008, 7, 3129.
5. Dai, Y.; Hartandi, K.; Ji, Z.; Ahmed, A. A.; Albert, D. H.; Bauch, J. L.; Bouska, J. J.;
Bousquet, P. F.; Cunha, G. A.; Glaser, K. B.; Harris, C. M.; Hickman, D.; Guo, J.; Li,
J.; Marcotte, P. A.; Marsh, K. C.; Moskey, M. D.; Martin, R. L.; Olson, A. M.;
Osterling, D. J.; Pease, L. J.; Soni, N. B.; Stewart, K. D.; Stoll, V. S.; Tapang, P.;
Reuter, D. R.; Davidsen, S. K.; Michaelides, M. R. J. Med. Chem. 2007, 50, 1584.
6. Potashman, M. H.; Bready, J.; Coxon, A.; DeMelfi, T. M., Jr.; DiPietro, L.; Doerr,
N.; Elbaum, D.; Estrada, J.; Gallant, P.; Germain, J.; Gu, Y.; Harmange, J.-C.;
Kaufman, S. A.; Kendall, R.; Kim, J. L.; Kumar, G. N.; Long, A. M.; Neervannan, S.;
Patel, V. F.; Polverino, A.; Rose, P.; Van der Plas, S.; Whittington, D.; Zanon, R.;
Zhao, H. J. Med. Chem. 2007, 50, 4351.
7. Ramurthy, S.; Subramanian, S.; Aikawa, M.; Amiri, P.; Costales, A.; Dove, J.;
Fong, S.; Jansen, J. M.; Levine, B.; Ma, S.; McBride, C. M.; Michaelian, J.; Pick, T.;
Poon, D. J.; Girish, S.; Shafer, C. M.; Stuart, D.; Sung, L.; Renhowe, P. A. J. Med.
Chem. 2008, 51, 7049.
8. Jung, M.-H.; Kim, H.; Choi, W.-K.; El-Gamal, M. I.; Park, J.-H.; Yoo, K. H.; Sim, T.
B.; Lee, S. H.; Baek, D.; Hah, J.-M.; Cho, J.-H.; Oh, C.-H. Bioorg. Med. Chem. Lett.
2009, 19, 6538.
9. Ménard, D.; Niculescu-Duvaz, I.; Dijkstra, H. P.; Niculescu-Duvaz, D.;
Suijkerbuijk, B. M.; Zambon, A.; Nourry, A.; Roman, E.; Davies, L.; Manne, H.
A.; Friedlos, F.; Kirk, R.; Whittaker, S.; Gill, A.; Taylor, R. D.; Marais, R.; Springer,
C. J. J. Med. Chem. 2009, 52, 3881.
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Process Res. Dev. 2002, 6, 777.
C
26H25ClF3N4O2S [M+H]+ 549.1334, found 549.1331. HPLC
purity = 97.9%.
5.3.38. 4-(4-(3-(4-Methoxyphenyl)thioureido)phenoxy)-N-
methylpicolinamide (9s)
White solid (yield 69%), mp: = 174–176 °C; 1H NMR (600 MHz,
DMSO-d6)
d 2.79 (d, J = 4.7 Hz, 3H), 3.75 (s, 3H), 6.92 (d,
J = 8.8 Hz, 2H), 7.17 (dd, J = 5.6, 2.5 Hz, 1H), 7.20 (d, J = 8.7 Hz,
2H) 7.33 (d, J = 8.8 Hz, 2H), 7.41 (d, J = 2.4 Hz, 1H), 7.58 (d,
J = 8.7 Hz, 2H), 8.52 (d, J = 5.6 Hz, 1H), 8.80 (br d, J = 4.8 Hz ,1H),
9.72 (br d, 2H); 13C NMR (300 MHz, DMSO-d6) d 26.96, 56.19,
109.81, 114.98, 115.14, 121.82, 126.64, 127.00, 133.02, 138.24,
150.47, 151.36, 153.43, 157.58, 164.71, 166.66, 180.94;
HRMS(AP-ESI) m/z: calcd for C21H21N4O3S [M+H]+ 409.1329, found
409.1333. HPLC purity = 98.63%.
5.3.39. N-Benzyl-4-(4-(3-(4-methoxyphenyl)thioureido)
zphenoxy)picolinamide (9t)
White solid (yield 69.4%), mp: = 148–152 °C; 1H NMR (600 MHz,
DMSO-d6) d 3.75 (s, 3H), 4.46 (d, J = 6.6 Hz, 2H), 6.91–6.93 (m, 2H),