Synthesis and evaluation of 4-(2-pyrimidinylamino) benzamides inhibitors of hedgehog signaling pathway

Article abstract of DOI:10.1016/j.bmcl.2013.12.050

A novel series of hedgehog signaling pathway inhibitors has been designed based on the 4-(2-pyrimidinylamino) benzamides scaffold. The synthesis and SAR of these compounds are described. Optimization leads to the identification of compound 3c, a potent and orally available agent with improved physicochemical and pharmacokinetic properties.

Full text of DOI:10.1016/j.bmcl.2013.12.050

Bioorganic & Medicinal Chemistry Letters 24 (2014) 983–988
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
Synthesis and evaluation of 4-(2-pyrimidinylamino) benzamides
inhibitors of hedgehog signaling pathway
b
b
b
b
b
b
Minhang Xin a,b, , Jun Wen , Feng Tang , Chongxing Tu , Wei Huang , Han Shen , Xinge Zhao ,
⇑
Lingfei Cheng b, Mengyu Wang b, Liandi Zhang b
a Department of Medicinal Chemistry, School of Pharmacy, Health Science Center, Xi’an Jiaotong University, No. 76, Yanta West Road, Xi’an 710061, P.R. China
b Jiangsu Simcere Pharmaceutical Co. Ltd., Jiangsu Key Laboratory of Molecular Targeted Antitumor Drug Research, No 699-18, Xuan Wu District, Nanjing 210042, P.R. 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 hedgehog signaling pathway inhibitors has been designed based on the 4-(2-pyrimidi-
nylamino) benzamides scaffold. The synthesis and SAR of these compounds are described. Optimization
leads to the identification of compound 3c, a potent and orally available agent with improved physico-
chemical and pharmacokinetic properties.
Received 18 September 2013
Revised 10 November 2013
Accepted 12 December 2013
Available online 17 December 2013
Ó 2014 Elsevier Ltd. All rights reserved.
Keywords:
Hedgehog signaling pathway
Inhibitors
SAR
Pharmacokinetic properties
The Hedgehog (Hh) signaling pathway is receiving increasing
attention because of its very role in malignant transformation of
various cancers.1 Genetic mutations associated with this signaling
pathway such as Ptch-mutation, Smo-mutation, and Sufu-
mutation result in tumorigenesis in several cancers such as basal
cell carcinoma (BCC), medulloblastoma, and rhabdomyosarcoma.
Additionally, strong evidences link overexpression of Hh ligands
to tumor aggravation of various other cancers such as hepatocellu-
lar, pancreatic, gastric, colorectal, esophageal, lung, ovarian,
prostate, melanoma, glioblastoma, leukemia, lymphoma.2 Thus,
inhibition of Hh signaling pathway is thought to have considerable
potential for therapy of many cancer types.
the discovery of a novel series of 4-(2-pyrimidinylamino) benzam-
ides as potent Hh signaling pathway inhibitors, exemplified by 1
(Fig. 2), which inhibited the Hh signaling pathway in Gli-luciferase
reporter assay with IC50 = 1.3 nM, more potent than control posi-
tive drug vismodegib. Initial SAR studies in this series showed that
structural modification on A-ring of 1 seriously affected the
inhibitory activity and the optimal substituents was conferred to
4-trifluoromethylphenyl, phenyl, 4-pyridinyl or others. Following
SAR studies focused on D-ring modifications, especially at the
position of phenyl group, demonstrating that methyl substitution
at C-2 of the phenyl or several basic side chains functionalized at
C-5 was favorable for high inhibitory activity.7
To date, many classes of Hh pathway inhibitors have been
described.3 Vismodegib has been recently approved by FDA for
metastatic BCC treatment.4 Other molecules including sonidegib
(NVP-LDE225, Phase III), LY-2940680 (Phase II), BMS-833923
(XL-139, Phase II), PF-04449913(Phase I), TAK-441 (Phase I),
NVP-LEQ506 (Phase I) have been already studied in clinic for many
tumors treatment (Fig. 1).5 Besides, there are still some novel small
molecules investigated in preclinical stage or drug discovery
stage.6
As a continuation of our study toward identification of potent
Hh signaling pathway inhibitors, a further SAR study was con-
ducted to explore various basic amine chains linked to D-ring,
looking to identify potent inhibitors with improved physicochem-
ical properties which was of importance for successfully develop-
ing drug candidate. Furthermore, considering that 1 exhibited
unsatisfactory pharmacokinetic (PK) profile, efforts to improve
the PK properties of this scaffold was also made by modification
on both A-ring and D-ring. Herein, in this report we disclose the
synthesis and SAR analysis of these analogs of Hh signaling path-
way inhibitors, and also we report the PK evaluation in vivo.
On the basis of the previous SAR study, at the outset, our strat-
egy focused on bearing various basic substitutions at C-5 of D-ring.
A number of analogs incorporating basic amine side chains at C-5
of D-ring were summarized in Table 1. The Hh signaling pathway
inhibitory activity of compounds 2a–2zj were evaluated by a
In aspiration of interest in developing Hh signaling pathway
inhibitors for treatment of many tumors, we recently detailed
⇑
Corresponding author. address: Department of Medicinal Chemistry, School of
Pharmacy, Health Science Center, Xi’an Jiaotong University, No. 76, Yanta West
Road, Xi’an 710061, P.R. China. Tel.: +86 25 85560000 3192.
E-mail address: xmhcpu@163.com (M. Xin).
0960-894X/$ - see front matter Ó 2014 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.bmcl.2013.12.050

984
M. Xin et al. / Bioorg. Med. Chem. Lett. 24 (2014) 983–988
Cl
H
O
Cl
O
N
N
H
N
N
N
H
N
N
H
O
O
O
CF3
N
S
O
N
N
H
O
vismodegib
(GDC-0449)
sonidegib
(NVP-LDE-225;LDE-225)
BMS-833923 (XL-139)
Exelixis /BMS Ph II
Curis/Genentech(Roche) launched
Novartis Ph III
O
N
N
O
O
O
N
N
N N
N
N
N
NH
N
N
HN
N
O
N
N
N
H
OH
NH
O
O
F
N
N
N
OH
CF3
CF3
CN
LY-2940680
Lilly Ph II
NVP-LEQ506
Novartis Ph I
PF-04449913
Pfizer Ph I
TAK-441
Millennium /Takeda Ph I
Figure 1. Chemical structures of clinical Hh signaling pathway inhibitors.
CF3
Steric hindrance could also affect the activity, for instance com-
pound 2zc bearing bulky dicyclohexylamine group, which
O
a
showed moderate activity. Besides, secondary amines like ethyla-
mine (2zd), hydroxyethylamine (2ze), tert-butylamine (2zf) or
cyclopentylamine (2zg) were tolerated, displaying nearly compa-
rable activity to lead compound 1, except 2zh that showing weak
potency of Hh inhibitory activity, which was considered due to
its length and extra basicity. In addition, interestingly, the inhibi-
tory activity of the basic amine substituents containing aromatic
groups such as furylmethamine (2zi) and phenylamine (2zj) were
retained, and these results were consistent with the above
reported ones 2q and 2r. In a word, this SAR study showed 30
compounds displayed better inhibitory activities than positive
control GDC-0449, of which 20 exhibited superior activities to lead
compound 1.
A
3
4
5
O
2
D
N
N
H
1
B
N
C
6
N
H
Figure 2. Structure of compound 1.
luciferase reporter in NIH3T3 cell carrying a stably transfected Gli-
reporter construct (Gli-luciferase reporter cell lines).7,8 Table 1 out-
lined the in vitro IC50 for this series of various basic substitutions at
C-5 of D-ring of 4-(2-pyrimidinylamino) benzamides9. Notably, all
the analogs bearing basic amine on the molecules (2a–2zj) showed
good Hh signaling inhibitory activity with IC50 varied from 0.23 to
35.35 nM. Of all the amines explored, some cyclic aliphatic amines,
such as morpholine (2a), dimethyl morpholine (2b), piperidine
(2c), thiomorpholine (2d), pyrrolidine (2e), were more effective
than the lead compound 1 and 7–20 fold higher activity than
GDC-0449. However the similar cyclic amine piperazine (2f)
showed significantly decreased activity, perhaps due to relatively
strong basicity of second basic center of NH. Other piperazine
derivatives such as N-methyl piperazine (2g), N-ethyl piperazine
(2h), N-phenyl piperazine (2i), N-cyclopropanecarbonyl piperazine
(2j) and piperazin-3-one (2k) exhibited improved potency com-
pared to 2f because of relatively suitable basicity. A survey of
substituted piperidines was investigated and led to some more
favorable piperidine derivatives including 4-hydroxy piperidine
(2l), piperidin-4-one (2m), tetramethyl piperidine (2n), and tetra-
methyl piperidin-4-one (2o). However, 4-piperidyl piperidine
(2p) displayed lower activity, while the activity of piperidine fusing
aromatic ring such as tetrahydroquinoline (2q) and tetrahydrothi-
enopyridine (2r) almost retained. These results indicated that add-
ing extra basicity was not favorable to the activity. The same case
was accounted by pyrrolidine derivatives 2s whose activity was
weaker than 2e. Additionally, two bicyclic amine derivatives, 2t
and 2u, were also investigated, and it was found 2t was 6-fold
more potent than 2u. After screening a variety of cyclic amine sub-
stituents, some acyclic amines were explored. It appeared a suit-
able length of acyclic amines was important for maintaining
activity. For examples, 2v–2z displayed subnanomolar inhibition.
However, compounds 2za and 2zb, displayed much lower potency,
with IC50 = 35.3 and 10.9 nM, respectively, mainly due to the larger
size of amine chains. In fact, the extra basicity of these two com-
pounds also strongly effected the decreased activity additively.
To evaluate whether these compounds had an improved
druggability compared to 1, of our most potent inhibitors, two
compounds 2a and 2b were further investigated by profiling iv
and po pharmacokinetics in SD rat.10 The results were illustrated
in Table 3. After intravenous injection with 1 mg/kg in SD rats,
the area-under-curve (AUC) of 2a reached 1.3 h
lg/mL, larger than
that of 1 (0.92 h g/mL). Such more plasma exposure is due part to
l
the much smaller volume of distribution (7.4 L/kg) and lower sys-
temic clearance (12.0 mL/min/kg). Furthermore, when orally
administered at 5 mg/kg dose, 2a showed higher oral exposure
and lower systemic clearance. Consequently, compound 2a
showed superior oral bioavailability (F = 46%) to 1 (36%). However,
unfortunately, 2b bearing N-methylpiperizine group showed poor
PK characteristics both by iv and po administration. Despite 2a
and 2b contained much hydrophilic group morpholine and N-
methylpiperizine in structures respectively, both molecules had al-
ready possessed higher lipophilicity (2a, AlogP = 7.07 and 2b,
AogP = 7.34) compared to vismodegib (AogP = 4.265). It appeared
attractive to attempt a structural modification to decrease the
AlogP value so as to further improve drug-like properties.
Therefore, Three compounds (3a–3c) were synthesized (Table 2),
beginning to replace of trifluoromethoxyphenyl of A-ring with
phenyl, 4-pyridinyl and 1-methylpyrazolyl because these groups
were previously reported to afford good to moderate inhibitory
activity.7 Of the three compounds, both compounds 3a and 3b
showed nearly equipotent inhibition with 2a in Hh signaling path-
way and possessed a lower AlogP values (3a, AlogP = 4.95 and 3b,
AogP = 4.069), but did not improve the plasma exposure in an
intravenous administration (Tables 2 and 3). Fortunately, at this
point, 3c with 1-methylpyrazolyl, showing equipotent potency
with 1 and a much lower AlogP (3c, AlogP = 3.6), was found to

M. Xin et al. / Bioorg. Med. Chem. Lett. 24 (2014) 983–988
985
Table 1
Table 2
In vitro inhibition of Hh signaling pathway for compounds modification at C-5 of D-
ring of 4-(2-pyrimidinylamino) benzamides
The IC50 and the AlogP values for compounds modification on A and D region of 4-(2-
pyrimidinylamino) benzamides9
R1
N
O
OCF3
R
N
N
H
3a-3k
O
N
H
2a-2zj
R
N
N
H
Gli-luc reporter IC50 (nM) ALogPa
N
N
H
Compds
R1
R
2a
2b
0.53
0.45
7.07
7.34
Compds
R
Gli-luc
reporter
IC50
Compds
R
Gli-luc
reporter
IC50
N
N
N
O
N
O
3a
3b
0.68
0.61
4.95
(nM)
(nM)
N
4.069
N
N
N
N
O
O
2a
2b
0.53
0.73
2t
1.6
N
N
3c
3d
3e
3f
1.3
3.6
N
N
2u
10.5
N
N
N
N
10.0
3.87
4.373
4.829
N
N
N
N
N
2c
2d
2e
2f
1.0
0.34
0.43
10.5
2v
2w
2x
2y
0.93
0.89
0.59
0.23
N
9.2
5.6
S
N
N
N
N
N
N
N
N
N
N
N
O
3g
8.4
4.355
N
N
NH
N
N
N
OH
OH
OH
3h
3i
21.2
17.5
3.345
4.785
2g
2h
2i
0.45
0.77
2.0
2z
0.47
35.3
10.9
N
N
N
N
N
N
N
N
2za
2zb
N
N
N
N
N
OH
OH
3j
31.9
18.1
2.833
4.308
N
H
N
N
HN
3k
O
N
N
2j
1.3
2.0
2zc
2zd
2.7
1
1.3
7.2
7.795
4.265
N
GDC-0449
O
NH
NH
a
ALogP values were calculated by Discovery studio 3.0 using Calculate Molecular
Properties Protocol.
2k
0.72
N
N
N
NH
OH
O
OH
2l
0.44
0.28
2ze
2zf
1.1
prepared (Table 2). Unfortunately, although 3d–3k also demon-
strated low AlogP values, only displayed less potency (5.6–
31.9 nM) compared to 1, which stalled their progression of further
PK studies. Therefore, based on the optimal studies of combination
of activity and PK properties, compounds 3c was considered to be
the ideal lead candidate, which exhibited superior potency, excel-
lent oral exposure and improved oral bioavailability in comparison
to 1. And now 3c were being evaluated in vivo in Hh pathway
dependent medulloblastoma allograft and other Hh signaling oper-
ative solid tumor models. The results will be reported later.
The 4-(2-pyrimidinylamino) benzamides were readily prepared
according to Scheme 1. Briefly, commercially available 2,4-di-
chloro-5-methylpyrimidine were treated with several aryl boronic
acid or boronic ester using palladium-catalyzed suzuki coupling
reaction to provide 4-aryl-substituted pyrimidines 5a–5d. The
intermediates 5a–5d were reacted with methyl 4-aminobenzoate
under palladium catalysis to afford the buckwald coupling prod-
ucts 6a–6d, which were further hydrolyzed to produce the free
acids 7a–7d. The condensation of 7a–7d with 3-amino-4-methyl-
benzyl alcohol, followed by chlorination and next acylation with
various amines, afforded the designed inhibitors 2a–2zj and
3a–3k (Scheme 1).7
HN
HN
2m
0.90
2n
0.76
2zg
1.6
N
2o
2p
2q
0.29
19.5
2.4
2zh
2zi
2zj
33.6
2.5
N
N
H
N
N
O
O
H
N
N
HN
1.1
N
S
2r
2s
1.5
3.9
1
1.3
7.2
N
N
GDC
N
significantly improve the oral exposure (AUC = 3.8 h
l
g/mL) by po
5 mg/kg dosage, moreover whose oral bioavailability reached al-
most 64%. In addition to the significant insight of low AlogP value
gained with the 1-methylpyrazolyl replacement, an additional
structure modification at position 5 of D-ring was pursued by
incorporating substituents, such as methylpiperizine, pyrrolidine,
piperidine, dimethylmorpholine, etc., which were identified to
have optimized activity above, then compounds 3d–3k were
In summary, we have utilized 4-(2-pyrimidinylamino) benzam-
ide core to produce novel Hh signaling pathway inhibitors. Starting
from the lead compound 1, we were able to explore structural
modification on A-ring and D-ring to identify potent inhibitors

986
M. Xin et al. / Bioorg. Med. Chem. Lett. 24 (2014) 983–988
Table 3
In vivo SD rat pharmacokinetic properties for represented compounds
compds
Dose (mg/kg)
Cmax
(lg/mL)
AUC (hÃ
lg/mL)
Vss (L/kg)
Cl (mL/min/kg)
MRT(0–t) (h)
T1/2 (h)
F (%)
GDC-0449
1
1 (iv)a
1 (iv)a
5 (po)a
1 (iv)a
5 (po)b
1 (iv)a
5 (po)b
1 (iv)a
1 (iv)a
1 (iv)a
5 (po)b
1.2
2.5
0.92
1.7
1.3
2.4
0.58
1.3
0.71
0.27
1.2
0.95
14.9
27.7
7.4
16.3
20.0
42.9
2.1
6.8
16.7
46.6
12.0
32.8
24.9
51.9
23.3
61.5
14.3
23.9
2.2
4.0
5.1
4.1
6.6
7.4
9.7
1.2
0.90
0.81
2.4
1.6
10.3
7.0
7.2
5.8
9.2
9.5
1.0
1.1
0.8
1.4
0.76
0.34
1.2
0.28
0.30
0.094
0.84
0.58
2.1
36.1
46.3
44.4
2a
2b
3a
3b
3c
5.7
0.90
2.9
1.5
3.8
63.6
a
compound was formulated using 5%DMA + 5% Tween-80.
Compound was formulated using 0.5%HPMC + 0.3% Tween-80, pH = 2.8.
b
R1
N
Cl
R1
N
O
R1
N
O
c
d
a
b
N
N
N
OCH3
OH
O
Cl
N
Cl
N
H
6a-6d
N
N
H
4
7a-7d
5a-5d
R1
R1
N
R1
O
O
f
e
R
N
N
H
N
H
N
N
H
OH
Cl
N
N
N
N
H
N
N
H
H
2a-2zj;
3a-3k
9a-9d
8a-8d
Scheme 1. Reagents and conditions: (a) Pd(PPh3)2Cl2, TEA, DMF, H2O, 80 °C for 6–12 h, 4-trifluoromethoxyphenylboronic acid for 5a, 73%; phenylboronic acid for 5b, 89%;
pyridine-4-boronic acid for 5c, 78%; 1-methyl-1H-pyrazole-4-boronic acid pinacol ester for 5d, 43%; (b) Pd(OAc)2, BINAP, Cs2CO3, dioxane, 170 °C, microwave for 3 h, methyl
4-aminobenzoate, 43–94%; (c) NaOH, MeOH/H2O, reflux overnight, 85–100%; (d) 3-amino-4-methylbenzyl alcohol, HATU, DMF, DIPEA, 85 °C overnight, 38–94%; (e) SOCl2,
CH2Cl2, rt, 2 h, 64–89%; (f) RH, DMF, K2CO3, rt, overnight; from 9a to 2a–2zj; from 9b to 3a; from 9c to 3b; from 9d to 3c–3k; 43–89%.
Marshall, D.; Marsters, J. C., Jr.; Murray, L. J.; Qian, C.; Rubin, L. L.; Salphati, L.;
Stanley, M. S.; Stibbard, J. H.; Sutherlin, D. P.; Ubhayaker, S.; Wang, S.; Wong, S.;
Xie, M. Bioorg. Med. Chem. Lett. 2009, 19, 5576; (b) http://www.fda.gov/.
with improved physicochemical properties. From these com-
pounds, some structure–activity relationships were concluded
and the compound 3c was identified as a potent and orally vailable
Hh signaling inhibitor with favorable pharmacokinetic properties.
Nowadays, the in vivo biological evaluation and the safety investi-
gation of 3c is being conducted both in Hh pathway dependent
medulloblastoma allograft and Hh signaling operative solid tumor
models, and the progress will be reported in due course.
5. (a) Pan, W.; Wu, X.; Jiang, J.; Gao, W.; Wan, Y.; Cheng, D.; Han, D.; Liu, J.;
Englund, N. P.; Wang, Y.; Peukert, S.; Miller-Moslin, K.; Yaun, J.; Guo, R.;
Matsumoto, M.; Vattay, A.; Jiang, Y.; Tsao, J.; Fangxian, S.; Pferdekamper, A. C.;
Dodd, S.; Tuntland, T.; Wieslawa, M.; Kelleher, J. F.; Yao, Y.; Warmuth, M.;
Williams, J.; Dorsch, M. ACS Med. Chem. Lett. 2010, 1, 130; (b) Rocconi, R. P.;
Samant, L.; WO 2013/043255, 2013.; (b) Ohashi, T.; Oguro, Y.; Tanaka, T.;
Shiokawa, Z.; Tanaka, Y.; Shibata, S.; Sato, Y.; Yamakawa, H.; Hattori, H.;
Yamamoto, Y.; Kondo, S.; Miyamoto, M.; Nishihara, M.; Ishimura, Y.; Tojo, H.;
Baba, A.; Sasaki, S. Bioorg. Med. Chem. 2012, 20, 5507; (c) Munchhof, M. J.; Li, Q.;
Shavnya, A.; Borzillo, G. V.; Boyden, T. L.; Jones, C. S.; LaGreca, S. D.; Martinez-
Alsina, L.; Patel, N.; Pelletier, K.; Reiter, L. A.; Robbins, M. D.; Tkalcevic, G. T. ACS
Med. Chem. Lett. 2012, 3, 106; (d) Peukert, S.; He, F.; Dai, M.; Zhang, R.; Sun, Y.;
Miller-Mislin, K.; McEwan, M.; Lagu, B.; Wang, K.; Yusuff, N.; Bourret, A.;
Ramamurthy, A.; Vattay, A.; Wang, A.; Guo, R.; Yuan, J.; Green, J.; Williams, J.;
Buonamici, S.; Kelleher, J. F.; Dorsch, M. ChemMedChem 2013, 8, 1261.
6. (a) Guerlet, G.; Spangenberg, T.; Mann, A.; Faure, H.; Ruat, M. Bioorg. Med. Chem.
Lett. 2011, 21, 3608; (b) Malancona, S.; Altamura, S.; Filocamo, G.; Kinzel, O.;
Hernando, J. I.; Rowley, M.; Scarpelli, R.; Steinkühler, C.; Jones, P. Bioorg. Med.
Chem. Lett. 2011, 21, 4422; (c) Kinzel, O.; Alfieri, A.; Altamura, S.; Brunetti, M.;
Bufali, S.; Colaceci, F.; Ferrigno, F.; Filocamo, G.; Fonsi, M.; Gallinari, P.;
Malancona, S.; Hernando, J. I.; Monteagudo, E.; Orsale, M. V.; Palumbi, M. C.;
Pucci, V.; Rowley, M.; Sasso, R.; Scarpelli, R.; Steinkühler, C.; Jones, P. Bioorg.
Med. Chem. Lett. 2011, 21, 4429; (d) Brown, M. L.; Aaron, W.; Austin, R. J.;
Chong, A.; Huang, T.; Jiang, B.; Kaizerman, J. A.; Lee, G.; Lucas, B. S.; McMinn, D.
L.; Orf, J.; Rong, M.; Toteva, M. M.; Xu, G.; Ye, Q.; Zhong, W.; Degraffenreid, M.
R.; Wickramasinghe, D.; Powers, J. P.; Hungate, R.; Johnson, M. G. Bioorg. Med.
Chem. Lett. 2011, 21, 5206; (e) Ontoria, J. M.; Bufi, L. L.; Torrisi, C.; Bresciani, A.;
Giomini, C.; Rowley, M.; Serafini, S.; Bin, H.; Hao, W.; Steinkühler, C.; Jones, P.
Bioorg. Med. Chem. Lett. 2011, 21, 5274; (f) Muraglia, E.; Ontoria, J. M.; Branca,
D.; Dessole, G.; Bresciani, A.; Fonsi, M.; Giuliano, C.; Llauger, Bufi L.;
Monteagudo, E.; Palumbi, M. C.; Torrisi, C.; Rowley, M.; Steinkühler, C.; Jones,
P. Bioorg. Med. Chem. Lett. 2011, 21, 5283; (g) Wang, J.; Mook, R. A., Jr.; Lu, J.;
Gooden, D. M.; Ribeiro, A.; Guo, A.; Barak, L. S.; Lyerly, H. K.; Chen, W. Bioorg.
Med. Chem. 2012, 20, 6751; (h) Peukert, S.; Jain, R. K.; Geisser, A.; Sun, Y.; Zhang,
R.; Bourret, A.; Carlson, A.; Dasilva, J.; Ramamurthy, A.; Kelleher, J. F. Bioorg.
Med. Chem. Lett. 2009, 19, 328; (i) Schaefer, G. I.; Perez, J. R.; Duvall, J. R.;
Stanton, B. Z.; Shamji, A. F.; Schreiber, S. L. J. Am. Chem. Soc. 2013, 135, 9578.
7. Xin, M.; Wen, J.; Tang, F.; Tu, C.; Shen, H.; Zhao, X. Bioorg. Med. Chem. Lett. 2013,
23, 6777.
Acknowledgments
This work was supported by the National Major Science and
Technology Project of China (Innovation and Development of
New Drugs, No. 2011ZX09401-008). The authors thank Dr. Haop-
eng Sun of China Pharmaceutical University for the AlogP
calculation.
References and notes
1. (a) Li, Y.; Maitah, M. Y.; Ahmad, A.; Kong, D.; Bao, B.; Sarkar, F. H. Expert Opin.
Ther. Targets 2012, 16(1), 49; (b) Zeng, X.; Goetz, J. A.; Suber, L. M.; Scott, W. J.,
Jr.; Schreiner, C. M.; Robbins, D. J. Nature 2001, 411, 716; (c) Stone, D. M.;
Hynes, M.; Armanini, M.; Swanson, T. A.; Gu, Q.; Johnson, R. L.; Scott, M. P.;
Pennica, D.; Goddard, A.; Phillips, H.; Noll, M.; Hooper, J. E.; Sauvage, F.;
Rosenthal, A. Nature 1996, 384, 129; (d) Tian, H.; Callahan, C. A.; DuPree, K. J.;
Darbonne, W. C.; Ahn, C. P.; Scales, S. J.; Sauvage, F. Proc. Natl. Acad. Sci. U.S.A.
2009, 106, 4254; (e) Onishi, H.; Katano, M. Cancer Sci. 2011, 102, 1756.
2. (a) Rubin, L. L.; de Sauvage, F. J. Nat. Rev. Drug Disc. 2006, 5, 1026; (b) Ng, J. M.;
Curran, T. Nat. Rev. Cancer 2011, 11, 493; (c) Metcalfe, C.; de Sauvage, F. J. Cancer
Res. 2011, 71, 5057; (d) Merchant, A. A.; Matsui, W. Clin. Cancer Res. 2010, 16,
3130.
3. (a) Mahindroo, N.; Punchihewa, C.; Fujii, N. J. Med. Chem. 2009, 52, 3829; (a)
Hadden, M. K. Expert Opin. Ther. Pat. 2013, 23, 345.
4. (a) Robarge, K. D.; Brunton, S. A.; Castanedo, G. M.; Cui, Y.; Dina, M. S.;
Goldsmith, R.; Gould, S. E.; Guichert, O.; Gunzner, J. L.; Halladay, J.; Jia, W.;
Khojasteh, C.; Koehler, M. F.; Kotkow, K.; La, H.; Lalonde, R. L.; Lau, K.; Lee, L.;

M. Xin et al. / Bioorg. Med. Chem. Lett. 24 (2014) 983–988
987
8. Ohashi, T.; Oguro, Y.; Tanaka, T.; Shiokawa, Z.; Shibata, S.; Sato, Y.; Yamakawa,
H.; Hattori, H.; Yamamoto, Y.; Kondo, S.; Miyamoto, M.; Tojo, H.; Baba, A.;
Sasaki, S. Bioorg. Med. Chem. 2012, 20, 5496.
(s, 1H, NH), 8.52 (s, 1H, ArH), 7.92 (s, 4H, ArH), 7.85 (d, 2H, J = 8.0 Hz, ArH), 7.55
(d, 2H, J = 7.6 Hz, ArH), 7.29 (s, 1H, ArH), 7.21 (d, 1H, J = 7.6 Hz, ArH), 7.11 (d,
1H, J = 6.8 Hz, ArH), 3.52 (s, 2H, PhCH2N), 3.21 (m, 1H, CHN), 2.90 (m, 2H,
CH2N), 2.67 (m, 2H, NCH2), 2.52 (m, 4H, N(CH2)2), 2.26 (s, 3H, ArCH3), 2.20 (s,
3H, ArCH3), 1.95 (m, 4H, (CH2)2), 1.56 (m, 4H, (CH2)2), 1.36 (m, 2H, CH2) ppm,
HPLC: 98.1%; 2q: MS (ESI) m/z: 1/2[M+H]+ = 312.7. 1H NMR (400 M, DMSO-d6) d
9.98 (s, 1H, CONH), 9.64 (s, 1H, NH), 8.52 (s, 1H, ArH), 7.91(m, 6H, ArH), 7.54
(m, 2H, ArH), 7.24 (d, 2H, J = 14 Hz, ArH), 7.04 (s, 1H, ArH), 6.87 (m, 2H, ArH),
6.45 (m, 2H, ArH), 4.44 (s, 2H, PhCH2N), 2.72 (m, 2H, CH2N), 2.26 (s, 3H, ArCH3),
2.18 (s, 3H, ArCH3), 1.92 (m, 2H, PhCH2), 1.23 (m, 2H, CH2) ppm, HPLC: 98.3%;
2r: MS (ESI) m/z: [M+H]+ = 630.2. 1H NMR (400 M, DMSO-d6) d 9.97 (s, 1H,
CONH), 9.64 (s, 1H, NH), 8.52 (s, 1H, ArH), 7.92(s, 4H, ArH), 7.87 (d, 2H,
J = 8.0 Hz, ArH), 7.55 (d, 2H, J = 8.4 Hz, ArH), 7.35 (s, 1H, ArH), 7.25 (m, 2H, ArH),
7.13 (d, 1H, J = 7.6 Hz, ArH), 6.76 (d, 1H, J = 5.2 Hz, ArH), 3.64 (s, 2H, PhCH2N),
3.44 (s, 2H, NCH2), 2.80 (m, 2H, NCH2), 2.76 (m, 2H, ArCH2), 2.26 (s, 3H, ArCH3),
2.22 (s, 3H, ArCH3) ppm, HPLC: 99.5%; 2s: MS (ESI) m/z: [M+H]+ = 645.3. 1H
NMR (400 M, DMSO-d6) d 9.97 (s, 1H, CONH), 9.64 (s, 1H, NH), 8.52 (s, 1H, ArH),
7.92(s, 4H, ArH), 7.87 (d, 2H, J = 8.0 Hz, ArH), 7.55 (d, 2H, J = 8.4 Hz, ArH), 7.26
(s, 1H, ArH), 7.19 (d, 1H, J = 8.0 Hz, ArH), 7.07 (d, 1H, J = 7.6 Hz, ArH), 4.12 (d,
1H, J = 13.2 Hz, CH), 3.31 (s, 2H, PhCH2N), 3.24 (m, 2H, CH2), 2.80 (m, 2H, CH2),
2.60 (m, 2H, CH), 2.26 (s, 3H, ArCH3), 2.19 (s, 3H, ArCH3), 2.10 (m, 1H, CH2), 1.91
(m, 1H, CH2), 1.67 (m, 4H, CH2CH2), 1.62 (m, 4H, CH2CH2) ppm, HPLC: 97.1%;
2t: MS (ESI) m/z: [M+H]+ = 602.3. 1H NMR (400 M, DMSO-d6) d 10.02 (s, 1H,
CONH), 9.70 (s, 1H, NH), 8.54 (s, 1H, ArH), 7.95(s, 4H, ArH), 7.89 (d, 2H,
J = 8.8 Hz, ArH), 7.57 (d, 3H, J = 8.4 Hz, ArH), 7.38 (d, 1H, J = 7.2 Hz, ArH), 7.27 (d,
1H, J = 7.2 Hz, ArH), 3.57 (s, 2H, PhCH2N), 2.71 (m, 4H, CH2NCH2), 2.30 (s, 3H,
ArCH3), 2.28 (s, 3H, ArCH3) 1.71 (m, 4H, 2*CH2), 1.54 (m, 2H, CHCH), 1.44 (m,
2H, CH2) ppm, HPLC: 97.3%; 2u: MS (ESI) m/z: [M+H]+ = 617.3. 1H NMR (400 M,
DMSO-d6) d 9.99 (s, 1H, CONH), 9.65 (s, 1H, NH), 8.54 (s, 1H, ArH), 7.94(s, 4H,
ArH), 7.89 (d, 2H, J = 8.0 Hz, ArH), 7.57 (d, 2H, J = 8.0 Hz, ArH), 7.28 (s, 1H, ArH),
7.22 (d, 1H, J = 7.6 Hz, ArH), 7.10 (d, 1H, J = 7.2 Hz, ArH), 3.56 (s, 2H, PhCH2N),
2.84 (m, 2H, NCH2), 2.64 (m, 2H, NCH2), 2.22 (m, 13H, NCH3+ArCH3+CH2), 1.88
(m, 1H, CH2), 1.48 (m, 1H, CH2) ppm, HPLC: 95.3%; 2v: MS (ESI) m/z:
[M+H]+ = 564.3. 1H NMR (400 M, DMSO-d6) d 9.98 (s, 1H, CONH), 9.64 (s, 1H,
NH), 8.52 (s, 1H, ArH), 7.92 (s, 4H, ArH), 7.87 (d, 2H, J = 8.4 Hz, ArH), 7.55 (d, 2H,
J = 8.0 Hz, ArH), 7.28 (s, 1H, ArH), 7.21 (d, 1H, J = 8.0 Hz, ArH), 7.08 (d, 1H,
J = 7.6 Hz, ArH), 3.43 (s, 2H, PhCH2N), 2.30 (s, 2H, CH2N), 2.26 (s, 3H, ArCH3),
2.20 (s, 3H, ArCH3), 2.12 (s, 3H, NCH3), 1.48 (m, 2H, CH2), 0.85 (t, 3H, J = 7.6 Hz,
CH3) ppm, HPLC: 97.3%; 2w: MS (ESI) m/z: [M+H]+ = 564.3. 1H NMR (400 M,
DMSO-d6) d 10.00 (s, 1H, CONH), 9.67 (s, 1H, NH), 8.52 (s, 1H, ArH), 7.94(s, 4H,
ArH), 7.88 (d, 2H, J = 8.8 Hz, ArH), 7.56 (d, 2H, J = 8.4 Hz, ArH), 7.32 (s, 1H, ArH),
7.22 (d, 1H, J = 7.6 Hz, ArH), 7.12 (d, 1H, J = 7.2 Hz, ArH), 3.53 (s, 2H, PhCH2N),
2.91 (s, 1H, CHN), 2.26 (s, 3H, ArCH3), 2.22 (s, 3H, ArCH3), 2.06 (s, 3H, NCH3),
1.05 (d, 6H, (CH3)2) ppm, HPLC: 96.0%; 2x: MS (ESI) m/z: [M+H]+ = 536.3. 1H-
NMR (400 M, DMSO-d6) d 9.96 (s, 1H, CONH), 9.62 (s, 1H, NH), 8.51 (s, 1H, ArH),
7.92 (s, 4H, ArH), 7.85 (d, 2H, J = 8.4 Hz, ArH), 7.52 (d, 2H, J = 8.4 Hz, ArH), 7.29
(s, 1H, ArH), 7.19 (d, 1H, J = 8.0 Hz, ArH), 7.08 (d, 1H, J = 7.6 Hz, ArH), 3.39 (s, 2H,
PhCH2N), 2.26 (s, 3H, ArCH3), 2.21 (s, 3H, ArCH3), 2.17 (s, 6H, N(CH3)2) ppm,
HPLC: 99.6%; 2y: MS (ESI) m/z: [M+H]+ = 564.3. 1H NMR (400 M, DMSO-d6) d
9.98 (s, 1H, CONH), 9.64 (s, 1H, NH), 8.52 (s, 1H, ArH), 7.93 (s, 4H, ArH), 7.87 (d,
2H, J = 8.4 Hz, ArH), 7.55 (d, 2H, J = 8.4 Hz, ArH), 7.30 (s, 1H, ArH), 7.20 (d, 1H,
J = 7.6 Hz, ArH), 7.10 (d, 1H, J = 7.2 Hz, ArH), 3.50 (s, 2H, PhCH2N), 2.51 (m, 4H,
CH2NCH2), 2.26 (s, 3H, ArCH3), 2.20 (s, 3H, ArCH3), 0.99 (s, 6H, (CH3)2) ppm,
HPLC: 97.6%; 2z: MS (ESI) m/z: [M+H]+ = 596.3. 1H NMR (400 M, DMSO-d6) d
9.95 (s, 1H, CONH), 9.62 (s, 1H, NH), 8.52 (s, 1H, ArH), 7.92(s, 4H, ArH), 7.87 (d,
2H, J = 8.0 Hz, ArH), 7.55 (d, 2H, J = 8.4 Hz, ArH), 7.29 (s, 1H, ArH), 7.19 (d, 1H,
J = 7.6 Hz, ArH), 7.12 (d, 1H, J = 7.6 Hz, ArH), 4.32 (t, 1H, J = 4.8 Hz, OH), 3.61 (s,
2H, PhCH2N), 3.46 (m, 4H, 2*CH2O), 2.56 (t, 4H, J = 6.0 Hz, 2*CH2N), 2.26 (s, 3H,
ArCH3), 2.20 (s, 3H, ArCH3) ppm, HPLC: 96.3%; 2za:MS (ESI) m/z:
[M+H]+ = 607.3. 1H NMR (400 M, DMSO-d6) d 9.99 (s, 1H, CONH), 9.67 (s, 1H,
NH), 8.52 (s, 1H, ArH), 7.92(m, 4H, ArH), 7.87 (d, 2H, J = 8.4 Hz, ArH), 7.55 (d,
2H, J = 8.0 Hz, ArH), 7.32 (s, 1H, ArH), 7.22 (d, 1H, J = 7.6 Hz, ArH), 7.09 (d, 1H,
J = 7.6 Hz, ArH), 3.38 (s, 2H, PhCH2N), 2.82 (t, 2H, J = 7.6 Hz, CH2), 2.56 (s, 6H,
2*CH3), 2.41 (t, 2H, J = 7.2 Hz, CH2), 2.26 (s, 3H, ArCH3), 2.22 (s, 3H, ArCH3), 2.17
(s, 3H, CH3), 1.78 (m, 2H, CH2) ppm, HPLC: 99.0%; 2zb: MS (ESI) m/z:
[M+H]+ = 635.3. 1H NMR (400 M, DMSO-d6) d 9.99 (s, 1H, CONH), 9.63 (s, 1H,
NH), 8.52 (s, 1H, ArH), 7.92 (s, 4H, ArH), 7.88 (d, 2H, J = 8.8 Hz, ArH), 7.56 (d, 2H,
J = 8.4 Hz, ArH), 7.28 (s, 1H, ArH), 7.20 (d, 1H, J = 7.6 Hz, ArH), 7.10 (d, 1H,
J = 7.6 Hz, ArH), 3.69 (s, 2H, PhCH2N), 2.93 (m, 2H, 2*CHN), 2.50 (m, 2H,
2*CH2N), 2.26 (s, 3H, ArCH3), 2.20 (s, 3H, ArCH3), 0.94 (m, 12H, 4*CH3) ppm,
HPLC:95.0%; 2zc: MS (ESI) m/z: [M+H]+ = 672.4. 1H NMR (400 M, DMSO-d6) d
9.98 (s, 1H, CONH), 9.60 (s, 1H, NH), 8.52 (s, 1H, ArH), 7.92(s, 4H, ArH), 7.88 (d,
2H, J = 8.0 Hz, ArH), 7.55 (d, 2H, J = 7.6 Hz, ArH), 7.29 (s, 1H, ArH), 7.14 (m, 2H,
ArH), 3.67 (s, 2H, PhCH2N), 2.26 (s, 3H, ArCH3), 2.18 (s, 3H, ArCH3), 1.79 (m, 2H,
2*NCH), 1.69 (m, 8H, 4*(CH2)), 1.23 (m, 8H, 4*(CH2)), 1.23 (m, 4H, 2*(CH2))
ppm, HPLC:94.54%; 2zd: MS (ESI) m/z: [M+H]+ = 536.2. 1H NMR (400 M, DMSO-
d6) d 10.01 (s, 1H, CONH), 9.72 (s, 1H, NH), 8.53 (s, 1H, ArH), 7.94 (s, 4H, ArH),
7.88 (d, 2H, J = 8.4 Hz, ArH), 7.56 (m, 3H, ArH), 7.35 (d, 1H, J = 8.0 Hz, ArH), 7.29
(d, 1H, J = 8.0 Hz, ArH), 4.11 (s, 2H, PhCH2N), 2.97 (m, 2H, CH2N), 2.26 (s, 6H,
2*ArCH3), 1.21 (t, 3H, CH3) ppm, HPLC: 98.8%; 2ze: MS (ESI) m/z:
[M+H]+ = 552.3. 1H NMR (400 M, DMSO-d6) d 9.98 (s, 1H, CONH), 9.63 (s, 1H,
NH), 8.52 (s, 1H, ArH), 7.92 (s, 4H, ArH), 7.87 (d, 2H, J = 7.2 Hz, ArH), 7.55 (s, 2H,
ArH), 7.29 (s, 1H, ArH), 7.18 (s, 1H, ArH), 7.11 (s, 1H, ArH), 4.47 (s, 1H, OH), 3.68
(s, 2H, CH2O), 3.46 (s, 2H, PhCH2N), 2.57 (s, 2H, CH2N), 2.26 (s, 3H, ArCH3), 2.20
(s, 3H, ArCH3) ppm, HPLC: 95.6%; 2zf: MS (ESI) m/z: [M+H]+ = 564.3. 1H NMR
(400 M, DMSO-d6) d 9.98 (s, 1H, CONH), 9.65 (s, 1H, NH), 8.52 (s, 1H, ArH),
9. Purity of all final compounds was assessed by HPLC and determined to be >95%.
The structures were confirmed by 1H NMR and LC/MS. Analytic data of
represented compounds 2a: MS (ESI) m/z: [M+H]+ = 578.3. 1H NMR (400 M,
DMSO-d6) d 9.97 (s, 1H, CONH), 9.63 (s, 1H, NH), 8.52 (s, 1H, ArH), 7.92 (s, 4H,
ArH), 7.85 (d, 2H, J = 8.4 Hz, ArH), 7.53 (d, 2H, J = 8.0 Hz, ArH), 7.28 (s, 1H, ArH),
7.19 (d, 1H, J = 7.6 Hz, ArH), 7.09 (d, 1H, J = 7.6 Hz, ArH), 3.57 (s, 4H,
morpholine-H), 3.43 (s, 2H, PhCH2N), 2.36 (s, 4H, morpholine-H), 2.26 (s, 3H,
ArCH3), 2.20 (s, 3H, ArCH3) ppm, HPLC: 95.9%; 2b: MS (ESI) m/z:
[M+H]+ = 606.3. 1H NMR (400 M, DMSO-d6) d 10.01 (s, 1H, CONH), 9.78 (s,
1H, NH), 8.52 (s, 1H, ArH), 7.95 (s, 4H, ArH), 7.87 (d, 2H, J = 8.4 Hz, ArH), 7.58 (s,
1H, ArH), 7.55 (d, 2H, J = 8.4 Hz, ArH), 7.42 (d, 1H, J = 7.6 Hz, ArH), 7.36 (d, 1H,
J = 8.0 Hz, ArH), 4.27 (s, 2H, PhCH2N), 3.98 (m, 2H, CHOCH), 3.26 (d, 2H,
J = 7.6 Hz, CH2N), 2.68 (m, 2H, CH2N), 2.27 (s, 3H, ArCH3), 2.26 (s, 3H, ArCH3),
1.12 (s, 3H, CH3), 1.11 (s, 3H, CH3) ppm, HPLC: 95.7%; 2c: MS (ESI) m/z:
[M+H]+ = 576.3. 1H NMR (400 M, DMSO-d6) d 9.98 (s, 1H, CONH), 9.64 (s, 1H,
NH), 8.52 (s, 1H, ArH), 7.92 (s, 4H, ArH), 7.87 (d, 2H, J = 8.8 Hz, ArH), 7.55 (d, 2H,
J = 8.4 Hz, ArH), 7.25 (s, 1H, ArH), 7.20 (d, 1H, J = 7.6 Hz, ArH), 7.07 (d, 1H,
J = 7.6 Hz, ArH), 3.38 (s, 2H, PhCH2N), 2.32 (m, 4H, (CH2)2N), 2.26 (s, 3H, ArCH3),
2.20 (s, 3H, ArCH3), 1.48 (m, 4H, (CH2)2), 1.39 (m, 2H, CH2) ppm, HPLC: 99.5%;
2d: MS (ESI) m/z: [M+H]+ = 594.2. 1H NMR (400 M, DMSO-d6) d 9.97 (s, 1H,
CONH), 9.63 (s, 1H, NH), 8.52 (s, 1H, ArH), 7.92(s, 4H, ArH), 7.87 (d, 2H,
J = 7.2 Hz, ArH), 7.54 (d, 2H, J = 6.8 Hz, ArH), 7.27 (s, 1H, ArH), 7.20 (d, 1H,
J = 6.4 Hz, ArH), 7.07 (d, 1H, J = 6.4 Hz, ArH), 3.46 (s, 2H, PhCH2N), 2.60 (m, 8H,
(CH2)4), 2.25 (s, 3H, ArCH3), 2.20 (s, 3H, ArCH3) ppm, HPLC: 99.6%; 2e: MS (ESI)
m/z: [M+H]+ = 562.3. 1H NMR (400 M, DMSO-d6) d 9.98 (s, 1H, CONH), 9.63 (s,
1H, NH), 8.52 (s, 1H, ArH), 7.93 (s, 4H, ArH), 7.87 (d, 2H, J = 8.0 Hz, ArH), 7.55 (d,
2H, J = 8.0 Hz, ArH), 7.29 (s, 1H, ArH), 7.20 (d, 1H, J = 7.6 Hz, ArH), 7.09 (d, 1H,
J = 7.2 Hz, ArH), 3.55 (s, 2H, PhCH2N), 2.44 (m, 4H, pyrolidine-H), 2.26 (s, 3H,
ArCH3), 2.21 (s, 3H, ArCH3), 1.69 (m, 4H, pyrolidine-H) ppm, HPLC: 98.8%; 2f:
MS (ESI) m/z: [M+H]+ = 577.3. 1H NMR (400 M, DMSO-d6) d 10.00 (s, 1H, CONH),
9.66 (s, 1H, NH), 8.52 (s, 1H, ArH), 7.93(s, 4H, ArH), 7.88 (d, 2H, J = 8.4 Hz, ArH),
7.55 (d, 2H, J = 8.0 Hz, ArH), 7.27 (s, 1H, ArH), 7.21 (d, 1H, J = 7.6 Hz, ArH), 7.08
(d, 1H, J = 7.2 Hz, ArH), 3.45 (s, 2H, PhCH2N), 2.77 (m, 4H, N(CH2)2), 2.35 (m, 4H,
N(CH2)2), 2.26 (s, 3H, ArCH3), 2.20 (s, 3H, ArCH3) ppm, HPLC: 98.7%; 2g: MS
(ESI) m/z: [M+H]+ = 591.3. 1H NMR (400 M, DMSO-d6) d 9.96 (s, 1H, CONH), 9.62
(s, 1H, NH), 8.52 (s, 1H, ArH), 7.92 (s, 4H, ArH), 7.85 (d, 2H, J = 8.4 Hz, ArH), 7.53
(d, 2H, J = 8.4 Hz, ArH), 7.26 (s, 1H, ArH), 7.18 (d, 1H, J = 7.6 Hz, ArH), 7.05 (d,
1H, J = 7.6 Hz, ArH), 3.41 (s, 2H, PhCH2N), 2.33 (m, 8H, piperazine-H), 2.26 (s,
3H, ArCH3), 2.20 (s, 3H, ArCH3), 2.14 (s, 3H, NCH3) ppm, HPLC:99.2%; 2h: MS
(ESI) m/z: [M+H]+ = 605.3. 1H NMR (400 M, DMSO-d6) d 9.99 (s, 1H, CONH), 9.64
(s, 1H, NH), 8.52 (s, 1H, ArH), 7.92(s, 4H, ArH), 7.87 (d, 2H, J = 8.4 Hz, ArH), 7.55
(d, 2H, J = 8.0 Hz, ArH), 7.26 (s, 1H, ArH), 7.20 (d, 1H, J = 7.6 Hz, ArH), 7.07 (d,
1H, J = 7.6 Hz, ArH), 3.41 (s, 2H, PhCH2N), 2.36 (m, 10H, CH2N(CH2)4)N), 2.26 (s,
3H, ArCH3), 2.20 (s, 3H, ArCH3), 0.96 (t, 3H, J = 7.2 Hz, CH3) ppm, HPLC: 97.7%;
2i: MS (ESI) m/z: [M+H]+ = 653.3. 1H NMR (400 M, DMSO-d6) d 9.97 (s, 1H,
CONH), 9.65 (s, 1H, NH), 8.52 (s, 1H, ArH), 7.92(s, 4H, ArH), 7.87 (d, 2H,
J = 8.4 Hz, ArH), 7.55 (d, 2H, J = 8.0 Hz, ArH), 7.31 (s, 1H, ArH), 7.21 (m, 3H, ArH),
7.12 (t, 1H, ArH), 6.92 (d, 2H, J = 8.4 Hz, ArH), 6.76 (t, 1H, ArH), 3.50 (s, 2H,
PhCH2N), 3.12 (s, 4H, N(CH2)2), 2.50 (s, 4H, N(CH2)2), 2.26 (s, 3H, ArCH3), 2.22
(s, 3H, ArCH3)ppm, HPLC: 98.6%; 2j: MS (ESI) m/z: [M+H]+ = 645.3. 1H NMR
(400 M, DMSO-d6) d 9.98 (s, 1H, CONH), 9.65 (s, 1H, NH), 8.52 (s, 1H, ArH),
7.93(s, 4H, ArH), 7.87 (d, 2H, J = 8.0 Hz, ArH), 7.55 (d, 2H, J = 7.6 Hz, ArH), 7.30
(s, 1H, ArH), 7.22 (d, 1H, J = 7.6 Hz, ArH), 7.10 (d, 1H, J = 7.2 Hz, ArH), 3.66 (s, 2H,
PhCH2N), 3.47 (s, 4H, CH2NCH2), 2.40 (m, 4H, CH2NCH2), 2.26 (s, 3H, ArCH3),
2.21 (s, 3H, ArCH3), 1.94 (m, 1H, cyclopropyl-H), 1.48 (m, 2H, CH2), 0.70 (m, 4H,
cyclopropyl-H) ppm, HPLC: 96.6%; 2k: MS (ESI) m/z: [M+H]+ = 591.3. 1H NMR
(400 M, DMSO-d6) d 10.00 (s, 1H, CONH), 9.67 (s, 1H, NH), 8.52 (s, 1H, ArH),
7.93(s, 4H, ArH), 7.88 (d, 2H, J = 8.4 Hz, ArH), 7.55 (d, 2H, J = 8.0 Hz, ArH), 7.31
(s, 1H, ArH), 7.23 (d, 1H, J = 8.0 Hz, ArH), 7.10 (d, 1H, J = 7.6 Hz, ArH), 3.51 (s, 2H,
PhCH2N), 3.17 (s, 2H, NCH2), 2.80 (m, 2H, NCH2), 2.51 (s, 2H, NCH2), 2.26 (s, 3H,
ArCH3), 2.21 (s, 3H, ArCH3) ppm, HPLC: 95.1%;2l: MS (ESI) m/z: [M+H]+ = 592.3.
1H NMR (400 M, DMSO-d6) d 9.97 (s, 1H, CONH), 9.63 (s, 1H, NH), 8.52 (s, 1H,
ArH), 7.92 (s, 4H, ArH), 7.85 (d, 2H, J = 8.4 Hz, ArH), 7.55 (d, 2H, J = 8.4 Hz, ArH),
7.25 (s, 1H, ArH), 7.19 (d, 1H, J = 8.0 Hz, ArH), 7.06 (d, 1H, J = 7.6 Hz, ArH), 4.52
(m, 1H, CH), 3.39 (s, 2H, PhCH2N), 2.67 (t, 2H, piperidine-H), 2.26 (s, 3H, ArCH3),
2.19 (s, 3H, ArCH3), 2.07 (t, 2H, piperidine-H), 1.70 (m, 2H, CH2), 1.38 (m, 2H,
CH2) ppm, HPLC: 95.9%; 2m: MS (ESI) m/z: [M+H]+ = 590.3. 1H NMR (400 M,
DMSO-d6) d 9.99 (s, 1H, CONH), 9.67 (s, 1H, NH), 8.53 (s, 1H, ArH), 7.93 (s, 4H,
ArH), 7.88 (d, 2H, J = 8.0 Hz, ArH), 7.56 (d, 2H, J = 8.0 Hz, ArH), 7.35 (s, 1H, ArH),
7.25 (d, 1H, J = 7.6 Hz, ArH), 7.13 (d, 1H, J = 7.2 Hz, ArH), 3.59 (s, 2H, PhCH2N),
2.70 (t, 4H, piperidone-H), 2.36 (t, 4H, piperidone-H), 2.27 (s, 3H, ArCH3), 2.25
(s, 3H, ArCH3) ppm, HPLC: 95.3%; 2n: MS (ESI) m/z: [M+H]+ = 632.3. 1H NMR
(400 M, DMSO-d6) d 9.97 (s, 1H, CONH), 9.58 (s, 1H, NH), 8.52 (s, 1H, ArH), 7.92
(s, 4H, ArH), 7.87 (d, 2H, J = 8.0 Hz, ArH), 7.55 (d, 2H, J = 8.0 Hz, ArH), 7.37 (s, 1H,
ArH), 7.20 (d, 1H, J = 7.2 Hz, ArH), 7.13 (d, 1H, J = 7.6 Hz, ArH), 3.74 (s, 2H,
PhCH2N), 2.26 (s, 3H, ArCH3), 2.16 (s, 3H, ArCH3), 1.56 (s, 2H, CH2), 1.46 (s, 4H,
(CH2)2), 0.97 (s, 12H, 4*CH3) ppm, HPLC: 97.0%; 2o: MS (ESI) m/z:
[M+H]+ = 646.3. 1H NMR (400 M, DMSO-d6) d 9.94 (s, 1H, CONH), 9.58 (s, 1H,
NH), 8.51 (s, 1H, ArH), 7.92(s, 4H, ArH), 7.87 (d, 2H, J = 8.0 Hz, ArH), 7.54 (d, 2H,
J = 6.8 Hz, ArH), 7.28 (s, 1H, ArH), 7.18 (d, 1H, J = 6.4 Hz, ArH), 7.07 (d, 1H,
J = 6.8 Hz, ArH), 3.86 (s, 2H, PhCH2N), 2.39 (s, 4H, CH2COCH2), 2.25 (s, 3H,
ArCH3), 2.18 (s, 3H, ArCH3), 1.96 (s, 12H, (CH3)4) ppm, HPLC: 95.1%; 2p: MS
(ESI) m/z: [M+H]+ = 659.3. 1H NMR (400 M, DMSO-d6) d 9.98 (s, 1H, CONH), 9.63

988
M. Xin et al. / Bioorg. Med. Chem. Lett. 24 (2014) 983–988
7.92(s, 4H, ArH), 7.87 (d, 2H, J = 8.0 Hz, ArH), 7.55 (d, 2H, J = 8.4 Hz, ArH), 7.32
7.97 (s, 4H, ArH), 7.31 (s, 1H, ArH), 7.21 (d, 1H, ArH), 7.10 (d, 1H, ArH), 3.97 (s,
3H, ArCH3), 3.57 (s, 2H, PhCH2N), 2.46 (m, 4H, pyrolidine-H), 2.35 (s, 3H,
ArCH3), 2.23 (s, 3H, ArCH3), 1.70 (m, 4H, pyrolidine-H) ppm, HPLC: 98.8%; 3f:
MS (ESI) m/z: [M+H]+ = 496.3. 1H NMR (400 M, DMSO-d6) d 9.76 (s, 1H, CONH),
9.66 (s, 1H, NH), 8.39 (d, 2H, ArH), 8.12 (s, 1H, ArH), 7.97 (s, 4H, ArH), 7.28 (s,
1H, ArH), 7.21 (d, 1H, ArH), 7.09 (d, 1H, ArH), 3.96 (s, 3H, ArCH3), 3.34 (s, 2H,
PhCH2N), 2.50 (m, 4H, piperidine-H), 2.34 (s, 3H, ArCH3), 2.22 (s, 3H, ArCH3),
1.50 (m, 4H, piperidine-H), 1.40 (m, 2H, piperidine-H) ppm, HPLC: 98.4%; 3g:
MS (ESI) m/z: [M+H]+ = 526.3. 1H NMR (400 M, DMSO-d6) d 9.77 (s, 1H, CONH),
9.67 (s, 1H, NH), 8.40 (d, 2H, ArH), 8.13 (s, 1H, ArH), 7.97 (s, 4H, ArH), 7.29 (s,
1H, ArH), 7.23 (d, 1H, ArH), 7.10 (d, 1H, ArH), 3.98 (s, 3H, ArCH3), 3.58 (m, 2H,
CHOCH), 3.42 (s, 2H, PhCH2N), 2.71 (m, 2H, morpholine-H), 2.35 (s, 3H, ArCH3),
2.22 (s, 3H, ArCH3), 1.65 (m, 2H, morpholine-H), 1.04 (d, 6H, 2*CH3) ppm, HPLC:
98.4%; 3h: MS (ESI) m/z: [M+H]+ = 512.3. 1H NMR (400 M, DMSO-d6) d 9.79 (s,
1H, CONH), 9.67 (s, 1H, NH), 8.41 (d, 2H, ArH), 8.13 (s, 1H, ArH), 7.98 (s, 4H,
ArH), 7.27 (s, 1H, ArH), 7.22 (d, 1H, ArH), 7.08 (d, 1H, ArH), 4.56 (s, 1H, OH), 3.97
(s, 3H, ArCH3), 3.41 (m, 1H, CHO), 3.36(s, 2H, PhCH2N), 2.68 (m, 2H, piperidine-
H), 2.35 (s, 3H, ArCH3), 2.22 (s, 3H, ArCH3), 2.02 (m, 2H, piperidine-H), 1.71 (m,
2H, piperidine-H), 1.39 (m, 2H, piperidine-H) ppm, HPLC: 98.6%; 3i: LC-MS
(ESI) m/z: [M+H]+ = 484.3. 1H NMR (400 M, DMSO-d6) d 9.80 (s, 1H, CONH), 9.73
(s, 1H, NH), 8.40 (d, 2H, ArH), 8.12 (s, 1H, ArH), 7.97 (s, 4H, ArH), 7.57 (s, 1H,
ArH), 7.34 (d, 2H, ArH), 3.96 (s, 3H, ArCH3), 3.35 (s, 2H, PhCH2N), 2.65 (m, 2H,
CH2N), 2.34(s, 3H, ArCH3), 2.28 (s, 3H, CH3N), 1.28 (m, 2H, CH2), 0.89 (m, 3H,
CH3) ppm, HPLC: 99.2%; 3j: MS (ESI) m/z: [M+H]+ = 516.3. 1H NMR (400 M,
DMSO-d6) d 9.77 (s, 1H, CONH), 9.67 (s, 1H, NH), 8.40 (d, 2H, ArH), 8.12 (s, 1H,
ArH), 7.98 (m, 4H, ArH), 7.29 (s, 1H, ArH), 7.20 (d, 1H, ArH), 7.13 (d, 1H, ArH),
4.37 (t, 2H, 2*OH), 3.96 (s, 3H, ArCH3), 3.61 (s, 2H, PhCH2N), 3.46 (m, 4H,
2*CH2O), 3.17 (m, 4H, 2*CH2N), 2.34(s, 3H, ArCH3), 2.21 (s, 3H, ArCH3) ppm,
HPLC: 98.0%; 3k: MS (ESI) m/z: [M+H]+ = 512.3. 1H NMR (400 M, DMSO-d6) d
9.77 (s, 1H, CONH), 9.67 (s, 1H, NH), 8.40 (d, 2H, ArH), 8.12 (s, 1H, ArH), 7.95 (s,
4H, ArH), 7.33 (s, 1H, ArH), 7.17 (d, 1H, ArH), 7.13 (d, 1H, ArH), 3.96 (s, 3H,
ArCH3), 3.66 (s, 2H, PhCH2N), 2.30 (s, 3H, ArCH3), 2.20 (s, 3H, ArCH3), 1.11 (s,
9H, 3*CH3) ppm, HPLC: 96.9%.
(s, 1H, ArH), 7.19 (d, 1H, J = 7.6 Hz, ArH), 7.13 (d, 1H, J = 8.0 Hz, ArH), 3.67 (s, 2H,
PhCH2N), 2.26 (s, 3H, ArCH3), 2.19 (s, 3H, ArCH3), 1.11 (s, 9H, (CH3)3) ppm,
HPLC: 98.5%; 2zg: MS (ESI) m/z: [M+H]+ = 576.3. 1H NMR (400 M, DMSO-d6) d
9.99 (s, 1H, CONH), 9.65 (s, 1H, NH), 8.52 (s, 1H, ArH), 7.92(s, 4H, ArH), 7.88 (d,
2H, J = 8.4 Hz, ArH), 7.56 (d, 2H, J = 8.4 Hz, ArH), 7.29 (s, 1H, ArH), 7.19 (d, 1H,
J = 8.0 Hz, ArH), 7.12 (d, 1H, J = 7.6 Hz, ArH), 3.66 (s, 2H, PhCH2N), 3.00 (m, 1H,
CHN), 2.26 (s, 3H, ArCH3), 2.19 (s, 3H, ArCH3), 1.73 (m, 2H, CH2), 1.64 (m, 2H,
CH2), 1.46 (m, 2H, CH2), 1.34 (m, 2H, CH2) ppm, HPLC: 95.0%;2zh: MS (ESI) m/z:
[M+H]+ = 619.3. 1H NMR (400 M, DMSO-d6) d 9.98 (s, 1H, CONH), 9.64 (s, 1H,
NH), 8.52 (s, 1H, ArH), 7.92 (s, 4H, ArH), 7.87 (d, 2H, J = 8.4 Hz, ArH), 7.55 (d, 2H,
J = 8.0 Hz, ArH), 7.31 (s, 1H, ArH), 7.21 (d, 1H, J = 7.6 Hz, ArH), 7.11 (d, 1H,
J = 7.2 Hz, ArH), 3.72 (s, 2H, PhCH2N), 2.63 (t, 2H, CH2N), 2.40 (t, 2H, NCH2), 2.32
(m, 4H, N(CH2)2), 2.26 (s, 3H, ArCH3), 2.20 (s, 3H, ArCH3), 1.47 (m, 4H, (CH2)2),
1.36 (m, 2H, CH2) ppm, HPLC: 95.4%: 2zi: MS (ESI) m/z: [M+H]+ = 588.3. 1H
NMR (400 M, DMSO-d6) d 9.99 (s, 1H, CONH), 9.67 (s, 1H, NH), 8.53 (s, 1H, ArH),
7.93(s, 4H, ArH), 7.87 (d, 2H, J = 8.4 Hz, ArH), 7.65 (s, 1H, ArH), 7.55 (d, 2H,
J = 8.4 Hz, ArH), 7.41 (s, 1H, ArH), 7.26 (d, 1H, J = 7.6 Hz, ArH), 7.18 (d, 1H,
J = 8.0 Hz, ArH), 6.45 (s, 1H, furan-H), 6.42 (s, 1H, furan-H), 3.89 (s, 2H, CH2N),
3.85 (s, 2H, PhCH2N), 2.26 (s, 3H, ArCH3), 2.23 (s, 3H, ArCH3) ppm, HPLC: 97.6%;
2zj: MS (ESI) m/z: [M+H]+ = 584.3. 1H NMR (400 M, DMSO-d6) d 9.98 (s, 1H,
CONH), 9.65 (s, 1H, NH), 8.52 (s, 1H, ArH), 7.91(s, 4H, ArH), 7.87 (d, 2H,
J = 8.4 Hz, ArH), 7.55 (d, 2H, J = 8.0 Hz, ArH), 7.33 (s, 1H, ArH), 7.21 (d, 1H,
J = 8.0 Hz, ArH), 7.15 (d, 1H, J = 7.6 Hz, ArH), 7.02 (m, 2H, ArH), 6.56 (m, 2H,
ArH), 6.51 (m, 1H, ArH), 6.24 (brs, 1H, ArNH), 4.24 (s, 2H, PhCH2N), 2.26 (s, 3H,
ArCH3), 2.18 (s, 3H, ArCH3) ppm, HPLC: 98.8%; 3a: MS (ESI) m/z:
[M+H]+ = 494.3. 1H NMR (400 M, DMSO-d6) d 9.94 (s, 1H, CONH), 9.63 (s, 1H,
NH), 8.50 (s, 1H, ArH), 7.92 (m, 4H, ArH), 7.72 (d, 2H, J = 7.6 Hz, ArH), 7.55 (m,
3H, ArH), 7.28 (s, 1H, ArH), 7.21 (d, 1H, J = 7.6 Hz, ArH), 7.09 (d, 1H, J = 7.6 Hz,
ArH), 3.57 (m, 4H, morpholine-H), 3.43 (s, 2H, PhCH2N), 2.35 (m, 4H,
morpholine-H), 2.26 (s, 3H, ArCH3), 2.20 (s, 3H, ArCH3) ppm, HPLC: 96.8%;
3b: MS (ESI) m/z: [M+H]+ = 508.3. 1H NMR (400 M, DMSO-d6) d 10.06 (s, 1H,
CONH), 9.66 (s, 1H, NH), 8.78 (d, 2H, J = 5.2 Hz, ArH), 8.57 (s, 1H, ArH), 7.92(s,
4H, ArH), 7.70 (d, 2H, J = 5.2 Hz, ArH), 7.28 (s, 1H, ArH), 7.21 (d, 1H, J = 7.6 Hz,
ArH), 7.08 (d, 1H, J = 7.6 Hz, ArH), 3.44 (s, 2H, PhCH2N), 2.42 (m, 8H,
N(CH2)2(CH2)2N), 2.26 (s, 3H, ArCH3), 2.23 (s, 3H, ArCH3), 2.20 (s, 3H, NCH3)
ppm, HPLC: 97.5%; 3c: MS (ESI) m/z: [M+H]+= 413.2. 1H NMR (400 M, DMSO-d6)
d 9.73 (s, 1H, CONH), 9.55 (s, 1H, NH), 8.39 (s, 1H, ArH), 8.36 (s, 1H, ArH), 8.11
(s, 1H, ArH), 7.97 (m, 4H, ArH), 7.11 (s, 3H, ArH), 3.96 (s, 3H, ArCH3), 2.33 (s, 3H,
ArCH3), 2.18 (s, 6H, 2*ArCH3) ppm, HPLC: 98.4%; 3d: MS (ESI) m/z:
[M+H]+ = 511.3. 1H NMR (400 M, DMSO-d6) d 9.77 (s, 1H, CONH), 9.66 (s, 1H,
NH), 8.40 (d, 2H, ArH), 8.13 (s, 1H, ArH), 7.96 (s, 4H, ArH), 7.28 (s, 1H, ArH), 7.22
(d, 1H, ArH), 7.09 (d, 1H, ArH), 3.97 (s, 3H, ArCH3), 3.43 (s, 2H, PhCH2N), 2.52 (s,
3H, NCH3), 2.35 (m, 8H, piperazine-H), 2.22 (s, 3H, ArCH3), 2.17 (s, 3H, ArCH3)
ppm, HPLC: 99.4%; 3e: MS (ESI) m/z: [M+H]+ = 483.3. 1H NMR (400 M, DMSO-
d6) d 9.77 (s, 1H, CONH), 9.66 (s, 1H, NH), 8.40 (d, 2H, ArH), 8.13 (s, 1H, ArH),
10. Pharmacokinetic Studies in SD rat. Compounds were administered to SD rats at
a
dose of 1 mg/kg (in 5% DMA+ 5% Tween-80) for iv or 5 mg/kg
(0.5%HPMC+0.3% Tween-80, pH=2.8) for po administration, respectively.
After oral administration, blood samples collected for PK analyses (n = 3)
were centrifuged at 4000 rpm for 5 min at 4 °C. Plasma samples were analyzed
after protein precipitation with acetonitrile acidified with1% formic acid. LC/
MS/MS analysis of test compounds was performed under optimized conditions
to obtain the best sensitivity and selectivity of the analyte in selected reaction
monitoring mode (SRM) containing an internal standard. Plasma
concentration-time data were measured by a noncompartmental approach
using the software WinNonlin Enterprise, version 5.2 (Pharsight Co.,Mountain
View, CA).