Design, synthesis and biological evaluation of 4-aminopyrimidine-5-cabaldehyde oximes as dual inhibitors of c-Met and VEGFR-2

Article abstract of DOI:10.1016/j.bmc.2016.03.061

The synergistically collaboration of c-Met/HGF and VEGFR-2/VEGF leads to development of tumor angiogenesis and progression of various human cancers. Therefore, inhibiting both HGF/c-Met and VEGF/VEGFR signaling may provide a novel and effective therapeutic approach for treating patients with abroad spectrum of tumors. Toward this goal, we designed and synthesized a series of derivatives bearing 4-aminopyrimidine-5-cabaldehyde oxime scaffold as potent dual inhibitors of c-Met and VEGFR-2. The cell proliferation assay in vitro demonstrated most target compounds have inhibition potency both on c-Met and VEGFR-2 with IC₅₀values in nanomolar range, especially compound 14i, 18a and 18b. Based on the further enzyme assay in vitro, compound 18a was considered as the most potent one, the IC₅₀s of which were 210?nM and 170?nM for c-Met and VEGFR-2, respectively. Following that, we docked the compound 10 and 18a with the proteins c-Met and VEGFR-2, and interpreted the SAR of these analogs. All the results indicate that 18a is a dual inhibitors of c-Met and VEGFR-2 that holds promising potential.

Full text of DOI:10.1016/j.bmc.2016.03.061

Accepted Manuscript
Design, synthesis and biological evaluation of 4-Aminopyrimidine-5-cabalde-
hyde oximes as dual inhibitors of c-Met and VEGFR-2
Hao Qiang, Weijie Gu, DanDan Huang, Wei Shi, Qianqian Qiu, Yuxuan Dai,
Wenlong Huang, Hai Qian
PII:
S0968-0896(16)30221-8
DOI:
http://dx.doi.org/10.1016/j.bmc.2016.03.061
BMC 12910
Reference:
To appear in:
Bioorganic & Medicinal Chemistry
Received Date:
Revised Date:
Accepted Date:
3 March 2016
27 March 2016
31 March 2016
Please cite this article as: Qiang, H., Gu, W., Huang, D., Shi, W., Qiu, Q., Dai, Y., Huang, W., Qian, H., Design,
synthesis and biological evaluation of 4-Aminopyrimidine-5-cabaldehyde oximes as dual inhibitors of c-Met and
VEGFR-2, Bioorganic & Medicinal Chemistry (2016), doi: http://dx.doi.org/10.1016/j.bmc.2016.03.061
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Design, synthesis and biological evaluation of 4-Aminopyrimidine-5-cabaldehyde oximes as
dual inhibitors of c-Met and VEGFR-2
a
Hao Qiang , Weijie Gu , DanDan Huang ,Wei Shi , Qianqian Qiu , Yuxuan Dai , Wenlong
a
a,b
a
a
a
a, c
Huang *, Hai Qian
a, c∗
a
Center of Drug Discovery, State Key Laboratory of Natural Medicines, China Pharmaceutical
University, 24 Tongjiaxiang, Nanjing 210009, PR China.
b
Department of Medicinal Chemistry, Nanjing Sanhome Pharmaceutical Co.,Ltd, Nanjing 210018,
PR China
c
Jiangsu Key Laboratory of Drug Discovery for Metabolic Disease, China Pharmaceutical
University, 24 Tongjiaxiang, Nanjing 210009, PR China
Abstract: The synergistically collaboration of c-Met/HGF and VEGFR-2/VEGF leads to
development of tumor angiogenesis and progression of various human cancers. Therefore,
inhibiting both HGF/c-Met and VEGF/VEGFR signaling may provide a novel and effective
therapeutic approach for treating patients with abroad spectrum of tumors. Toward this goal, we
designed and synthesized a series of derivatives bearing 4-Aminopyrimidine-5-cabaldehyde oxime
scaffold as potent dual inhibitors of c-Met and VEGFR2. The cell proliferation assay in vitro
demonstrated most target compounds have inhibition potency both on c-Met and VEGFR2 with
IC50 values in nanomolar range, especially compound 14i, 18a and 18b. Based on the further
enzyme assay in vitro, compound 18a was considered as the most potent one, the IC50s of which
were 210nM and 170nM for c-Met and VEGFR2, respectively. Following that, we docked the
compound 10 and 18a with the proteins c-Met and VEGFR-2, and interpreted the SAR of these
analogues. , All the results indicate that 18a is a dual inhibitors of c-Met and VEGFR2 that holds
promising potential.
Key words: Oxime; c-Met; VEGFR-2; synergistically collaborate; cancers
1. Introduction
∗
Co-corresponding authors: Wenlong Huang and Hai Qian, Center of Drug Discovery, State Key Laboratory of
Natural Medicines, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing 210009, China. Tel:
86-25-83271302; Fax: +86-25-83271480.
E-mail: ydhuangwenlong@126.com (W.l. Huang), qianhai24@163.com (H. Qian)
+
1

Receptor tyrosine kinases (RTKs) play an important role in the regulation of intracellular
1
signal transduction. Aberrant RTKs signaling leads to broad spectrum of tumor. RTKs have
become important targets for cancer therapy, and more and more RTKs inhibitors have been
designed and synthesized.
The c-Met receptor tyrosine kinase and its natural ligand (HGF) are involved in embryonic
development, wound healing, and oncogenesis. c-Met or HGF overexpression, metagenomic
amplification, or c-Met mutations leads to aberrant expression of c-Met/HGF signal pathway
which induces proliferation, invasion, metastasis, tumor angiogenesis, and drug resistance of
2
cancer cell in broad spectrum of tumor. Thus, inhibition of c-Met signaling has been considered
,3
as an effective way for cancer therapy. c-Met TKIs for treatment of cancer patients have been
3
developed and evaluated in clinical trials. On the basis of action of c-Met and its inhibitor, the
-5
6-9
c-Met TKIs have been classified to type I and type II TKIs. Type II c-Met inhibitors can interrupt
6
, 9-12
c-Met and VEGFR-2 or other kinase signaling pathways
.
In addition, vascular endothelial growth factor receptor 2 (VEGFR2) is a tyrosine kinase
receptor expressed in endothelial cells. Once VEGF binding to VEGFR, a conformational change
in VEGFR occurred, followed by dimerization and phosphorylation of tyrosine kinase receptor.
The VEGF/VEGFR2 signaling pathway is believed to be a major driver of tumor angiogenesis,
which supplies oxygen and nutrients to promote tumor growth and plays important roles in tumor
7
,13-17
malignancy (such as sustaining tumor growth) and in blood-borne metastasis.
Its
overexpression has been reported to correlate with the degree of vascularity, aggressive disease,
and poor prognosis in the majority of human solid tumors, making the VEGF/VEGFR axis an
attractive molecular target for cancer therapy.
Based on the above-mentioned findings, we believe that the synergistically collaboration of
c-Met and VEGFR-2 promotes development of angiogenesis and progression of various human
11, 15, 18
cancers
. Therefore, molecules that dually inhibit c-Met and VEGFR-2 may have broader
advantages over either c-Met- or VEGFR-2 selective inhibitor, since they can inhibit multiple
signaling pathways involved in tumor angiogenesis, proliferation, and metastasis. Thus, the dual
inhibitors interrupt c-Met and VEGFR-2 signaling pathways represent a promising approach to
1
cancer treatment . Therefore, we initiate synthetic studies of dual inhibitor of c-Met and
9
VEGFR-2 with potent antitumor efficacy.
2

The lead compound with potent antitumor efficacy was designed by structure-based drug
design. At first, we analyzed an X-ray crystal structure of c-Met in complex with the type II c-Met
TKIs N-{4-[(6,7-dimethoxy-4-quinolyl)oxy]phenyl}-N-(2-phenylacetyl)thiourea(Kirin Brewery).
The crystal structure indicates that the c-Met protein includes a lipophilic back pocket region and
adopts an inactive DFG-out conformation, which is the binding mode of type II c-Met TKIs (PDB
code: 3VW8). The scaffold occupies hinge region which can combine with ATP. These
backgrounds lead us to design the type-II dual inhibitors with potent antitumor efficacy by using
series scaffolds of VEGFR-2 inhibitors. Compound I (in Figure 1), including quinazoline scaffold,
2
0
has been reported only as potent VEGFR-2 inhibitor , quinazoline can bound to hinge region by
hydrogen bond. 4-aminopyrimidine-5-carbaldehyde oxime is considered as isosteric structure of
quinazoline since the intramolecular hydrogen bond forming a pseudo six-membered ring (II in
Figure 1). A series of VEGFR-2 inhibitors with 4-aminopyrimidine-5-carbaldehyde oxime have
2
been described, exemplified by compound II and III (JNJ-38158471). Cabozantinib (in Figure
1
22
2
3
) and Foretinib (in Figure 1), compounds for treatment of medullary thyroid cancer, were
1
2
previously reported to inhibit c-Met and VEGFR-2 simultaneously. The difference biological
4
profiles of these compounds indicated that the cyclopropane-1,1-dicarboxamide moiety might be
2
5
critical for the potent dual inhibition of c-Met and VEGFR-2. These research in this area inspired
the design of first dual inhibitor 10 (in Figure1), which was a split structure containing both the
cyclopropane-1, 1-dicarboxamide moiety and the 4-aminopyrimidine-5-carbaldehyde oxime for
inhibiting c-Met and VEGFR-2. The total compounds contain solvent region, scaffold and side
chain region. We preliminary studied the SAR of solvent region and scaffold.
3

Figure 1. C-Met and/or VEGFR-2 dual inhibitor and the design of novel inhibitor for both c-Met and VEGFR-2
kinase
2
. Chemistry
The compound 1 was prepared by condensation of diethyl malonate and 1, 2-dibromoethane
under 50% sodium hydroxide solution. The compound 1 was coupled with equal equivalence
-fluoroaniline by utilizing sulfoxide chloride and triethylamine to obtain 2. On the other way, 3
4
was synthesized by benzyl alcohol and 3, 4-difluoronitrobenzene under basic condition at reflux
temperature. The amine of 4 was prepared by reduction of the nitro of 3. The compound 5 was
prepared by condensation of 2 and 4 under basic condition at room temperature. Under the
hydrogen atmosphere, the benzyl of 5 had been removed by hydrogenolysis.
4

2 2
Scheme 1. Reagents and conditions: a. Dibromoethane, TEBA, NaOH/H O, rt, 3h; b. 4-Fluoroaniline, SOCl ,
0
N, THF, 0 C -rt, 3h; c. Benzyl alcohol, KOH, CH
Et
3
3
CN, rt, overnight; d. iron powder, ammonium formate,
0
Toluene/H
2
2 3 2
O, reflux, 8h; e. compd. 2 and 4, SOCl , Et N, THF, 0 C, overnight; f. Pd/C, H , THF/MeOH, rt, 7h.
The synthesis of 4-aminopyrimidine-5-carbaldehyde oxime derivatives 10, 13, 14a-14i,
5a-15d, 16a-16b, 18a-18c were shown in Scheme 2. The compound 7 was synthesized by
1
3
vilsmeier reaction, the material 4, 6-dihydroxypyrimidine was heated with POCl and DMF, then
react with 6 under basic condition at reflux temperature to obtain compound 8 which can be
condensed in polar solvent to obtain 9. The 6-Cl of compound 9 was replaced by methylamine to
obtain the first inhibitor 10. On the other synthetic way, the compound 11 could be prepared by
mono-substituted reaction with the materials 7 and ammonia (2M solution in methanol), then, the
target compound 12 were obtained by a condensation reaction of 11 and 6. Compound 13 were
synthesized by utilizing hydroxylamine hydrochloride with compound 12. The target compounds
14a-14l were prepared by condensation reaction of the compound 13 and a series of bromides. To
obtain 15a-15d, 16a and 16b, we utilize compound 9 and series O-instead hydroxylamine or series
substituted hydrazine. The reaction of 10 with 1, 3-dibromopropane could produce compound 17
which could react with series primary amine to obtain 18a-18c.
5

0
0
Scheme 2. Reagents and conditions: a. POCl
Methoxyamine hydrochloride, DMSO; d. Methylamine hydrochloride, K
compd 6, K CO , DMF, rt, 3h; g. hydroxylamine hydrochloride, DMSO, 2h; h. RBr, K
R-O-NH .HCl, DMSO, overnight or R-NH-NH , EtOH, overnight; j. 1,3-Dibromopropane, K
NH, K CO , DMF.
3
, DMF, 0 C; 130 C, 5h; b. compd 6, DIEA, EA, 2h; c.
2
CO3. e. Toluene, NH in MeOH, rt; f.
3
2
3
2
CO , DMF; DMF i.
3
2
2
2 3
CO , DMF; k.
R
1
R
2
2
3
3
. Results and discussion
.1 Cell proliferation inhibition and SAR study of target compounds
The inhibitory activities of the compounds prepared against c-Met and VEGFR2 kinases
were determined by the elisa assay using anti-phosphotyrosine antibodies. The effects of the
compounds on cell proliferation were evaluated using BaF -TPR-Met cells, in which the
c-Met-dependent cell line, and in VEGF-stimulated human umbilical vein endothelial cells
HUVEC).
At first, we attempted to dock the first dual inhibitor 10 with the c-Met crystal structure
3
3
(
2
derived from the complex of c-Met and cabozantinib (PDB code: 3LQ8) and then with the
3
2
VEGFR-2 crystal structure (PDB code: 3U6J) by MOE v2008.10. We found that the pyrimidine
6
of 4-aminopyrimidine-5-carbaldehyde oxime could bound to the pocket of ATP in c-Met and
6

VEGFR-2 receptor proteins by molecular docking. Met1160 of c-Met hinge region and Cys919 of
VEGFR-2 hinge region can form hydrogen bonds with nitrogen-atoms of pyrimidine as shown in
Figure 2A (green lines). However, the compound 10 could not be stabilized in the binding pocket
of c-Met and VEGFR-2 because the loss of hydrogen bonds between 4-amino and residue Met1160
of c-Met and Cys919 of VEGFR-2 (Figure 2A). The free 4-amino of scaffold can bound to Met1160
and Cys919 by second hydrogen bonds as shown in Figure 2B(red lines). Therefore, the candidate
18a may have potent inhibitory.
As shown in Table 1, the 4-substituted amino derivative 10 was inactive against c-Met
kinase and VEGFR-2 kinase (IC50>10000nM). On the other hand, the unsubstituted derivative 18a
was found to display both c-Met (IC50=194nM) and VEGFR-2(IC50=260nM) (in Table 1)
inhibitory activities, as suggested by docking model studies shown potent inhibitory activity, as
suggested by the docking model studies.
Next, we focused on the modification of the substituent groups of scaffold to further enhance
c-Met/VEGFR2 kinase inhibitory activity. At first, the alkanes (14a-14c), olefins/alkynes (14d,
1
4e), naphthenic groups (14f, 14h) and benzyls (14g) were introduced to the solvent region. The
increase of carbon atoms of solvent region leads to the increase of Log P and weaks the inhibitory
activity for BaF -TPR-Met. Comparing the hydroxypropyl substituted compound (14i), the
3
hydroxyethyl substituted compound (15a) has poor activities. Because the methylation of
hydroxyl of compound 14j, the activity down from 0.11µM to 0.24µM. The compound 14k was
inactive for BaF3-TPR-Met since the introduction of ester group. The introduction of the fragment
of formylmorpholine generated general activity (14l). For reducing Log P, we introduced the
fragments of tertiary in solvent area which lead to the increase of activities (15c, 18a-18c).
However, the length of carbon chain has little effect on the activity. Dimethyl amino substituted
compound 18a(c-Met IC50=89nm; VEGFR-2 IC50=190nm) is superior to pyrrolidine substituted
(
15c, 18c) and morpholine ring substituted (18b) as shown in Table 1. When oxime was replaced
by pyridine-hydrazone, the activity was reduced significantly as shown in Table 1(16a, 16b).
Table 1: Structures, Log Ps and IC50s of target compounds against BaF3-TRP-Met in vitro
7

a
b
Compd.
R
1
R
2
IC50 (µmol/L
)
Log P
1
1
1
1
1
1
1
1
0
CH
3
O
CH
H
3
>1
3.52
2.62
3.04
3.41
4.20
3.65
3.34
4.27
3
HO
CH
0.194 ±0.031
0.166 ±0.038
0.215±0.012
0.626±0.024
0.619±0.025
0.521±0.041
0.745±0.016
4a
4b
4c
4d
4e
4f
3
O
H
CH
CH
CH
3
CH
(CH
=CHCH
2
O
H
3
2
)
2
CH
2
O
H
2
2
O
H
H
H
1
1
4g
4h
PhCH2O
H
H
0.566±0.060
0.879±0.020
4.45
4.69
a
1
1
1
4i
4j
4k
H
H
H
0.110±0.018
2.67
3.18
3.15
0.243±0.017
>1
1
4l
H
0.397±0.031
2.10
1
1
5a
5b
H
H
0.200±0.018
0.258±0.095
2.37
2.75
1
1
5c
5d
H
H
0.162±0.017
0.158±0.036
3.50
2.02
NH
2
H
H
0.765±0.046
0.321±0.028
2.25
3.70
1
1
6a
6b
8

a
1
1
8a
8b
H
H
0.089±0.005
3.23
3.82
0.119±0.058
1
8c
H
0.186±0.074
2.98
c
Foretinib
0.0092±0.00097
4.28
c
Cabozantinib
0.0219±0.00282
0.01986±0.00251
4.28
NT
c
Cabozantinib L-Malate
NT, not tested
a
Data presented is the mean ± SD value of three independent determinations.
LogP values were determined by shake-flask procedure.
Used as positive control.
b
c
A
B
Figure 2.A. the model of 10 bound to c-Met (PDB code: 3LQ8) and VEGFR-2(PDB code: 3U6J); B. the
model of 18a with c-Met (PDB code: 3LQ8) and VEGFR-2(PDB code: 3U6J)
3.2 Enzymatic activities of optimized compounds
After the determination of the compounds on cell proliferation on BaF
3
-TRP-Met and
HUVEC, the compounds 13, 14a, 14i, 15c, 15d, 18a, 18b, 18c were assayed with the enzymatic
9

2
2
23
activities against c-Met and VEGFR-2 by ELISA, using Cabozantinib and Foretinib as
comparisons (Table 2). The compound 18a displays a better enzymatic potency than other
compound on c-Met and VEGFR-2 kinase.
Table 2: VEGFR-2 kinase activity of selected compounds and enzymatic activities in vitro
HUVEC
c-Met
VEGFR-2
Compd.
a
a
a
IC50(µmol/L)
IC (µmol/L)
50
IC (µmol/L)
5
0
1
1
1
1
1
1
1
1
1
0
ND
ND
ND
3
0.260±0.072
0.320±0.21
0.210±0.18
0.168±0.116
0.720±0.20
0.270±0.055
0.380±0.005
0.280±0.024
0.230±0.073
0.980±0.074
0.210±0.030
0.920±0.017
0.420±0.21
0.0032±0.29
0.0032±0.40
ND
0.140±0.047
0.120±0.058
0.560±0.060
0.250±0.063
0.890±0.016
0.170±0.055
0.340±0.062
0.220±0.035
ND
4a
4i
5c
5d
8a
8b
8c
a
0.110±0.005
0.219±0.014
0.427±0.026
0.017±0.016
ND
b
Foretinib
b
a
Cabozantinib
0.003±0.2
b
Cabozantinib L-Malate
0.045±0.39
ND
ND, not determined
a
Data presented is the mean ± SD value of three independent determinations.
Used as positive control.
b
4. Conclusion
In conclusion, novel dual c-Met/VEGFR-2 receptor tyrosine kinase inhibitors based the
-aminopyrimidine-5-carbaldehyde oxime scaffold were designed and synthesized. The
4
derivatives 13, 14a, 14i, 15c, 15d, 18a, 18b, 18c contain different substituted oxime ether on
-position of 4-aminopyrimidine-5-carbaldehyde oxime scaffold have preferable activity.
5
Compound 18a with the tertiary amino fragment exhibits potent activities in cell proliferation and
enzyme assays in vitro. Derivative 10 has low inhibition at 1µM concentration, that indicates free
6-amino of pyrimidine ring is essential groups for inhibitory activities. The introduction of
10

pyrimidine hydrazone on pyrimidine ring weaken c-Met/VEGFR-2 inhibitory activities since the
-oxime ether is essential for activities. Our findings indicated that compound 18a could be a lead
molecule for the further research of dual inhibitor of c-Met and VEGFR2.
5
5
. Experimental section
.1 Chemistry
5
Melting points were determined on RY-1 melting point apparatus, and were not corrected.
1
Proton nuclear magnetic resonance ( H-NMR) spectra were recorded on Bruker ACF-300/500
MHz instruments. Chemical shifts are reported as δ values (ppm) downfield from internal
tetramethylsilane of the indicated organic solution. Peak multiplicities are expressed as follows: s,
singlet; d, doublet; t, triplet; q, quartet; dd , doublet of doublet; ddd , doublet of doublet of
doublets; br, broad singlet; m, multiplet. Coupling constants (J values) are given in hertz
(
(
Hz).Mass spectra(MS) were acquired using an Waters UPLC/MS(ACQUITY UPLC/TQD
Waters® UPLC® with the ACQUITY® TQD [Waters Corporation, Milford, MA, USA]))
operating in electron spray ionization mode (ESI+) and were used to confirm the purity of each
compound. TLCs and preparative TLC were performed on silica gel GF/UV 254. Purifications by
column chromatography were carried out over silica gel (200–300 mesh) and visualized under UV
light at 254 and 365 nm. Reagents and solvents were obtained from commercial sources and used
without further purification.
1 13
The physical characteristics, H-NMR, C-NMR, MS and elemental analysis data for all
intermediates and target compounds, were reported in the supporting information.
5.2 Cell Proliferation Assay
BaF3-TRP-Met and HUVECs were seeded in 96-well plates in RPMI1640 medium with 10%
fetal bovine serum (FBS) and human endothelial serum free medium (Invitrogen), respectively.
The following day, serial dilutions of compounds or 0.1% DMSO wereadded to each well.
BaF3-TRP-Met were then incubated for further 72 h.Proliferation of HUVECs was stimulated
with 60 ng/mL recombinant human VEGF (R&D Systems, Minneapolis, MN) for 120 h. After
incubation, cell proliferation was determined using Cell Counting Kit-8 (DOJINDO Laboratories,
11

Kumamoto, Japan). IC50 values werecalculated by nonlinear regression analysis using GraphPad
Prism
(
GraphPad Software, Inc.).
5.3 In vitro c-Met and VEGFR2 enzyme assay
The effects of the compounds on the activities of two tyrosine kinases were determined using
enzyme-linked immunosorbent assays (ELISAs) with purified recombinant proteins. Briefly, 20
µg/mL poly (Glu,Tyr)4:1 (Sigma) was pre-coated in 96-well plates as a substrate. A 50 µL aliquot
of 10 µmol/L ATP solution diluted in kinase reaction buffer (50 mmol/L HEPES [pH 7.4], 50
2 2 3 4
mmol/L MgCl , 0.5 mmol/L MnCl , 0.2 mmol/L Na VO , and 1 mmol/L DTT) was added to each
well; 1 µL of various concentrations of indicated compounds diluted in 1% DMSO (v/v) (Sigma)
were then added to each reaction well. DMSO (1%, v/v) was used as the negative control. The
kinase reaction was initiated by the addition of purified tyrosine kinase proteins diluted in 49µL
of kinase reaction buffer. After incubation for 60 min at 37 °C, the plate was washed three times
with phosphate-buffered saline (PBS) containing 0.1% Tween 20 (T-PBS). Anti-phosphotyrosine
(
PY99) antibody was then added. After a 30-min incubation at 37 °C, the plate was washed three
times, and horseradish peroxidase-conjugated goat anti-mouse IgG was added. The plate was then
incubated at 37 °C for 30 min and washed 3 times. A 100 µL aliquot of a solution containing 0.03%
H
2
O
2
and 2 mg/ml o-phenylenediamine in 0.1 mol/L citrate buffer (pH5.5) was added. The
reaction was terminated by the addition of 50 µL of 2 mol/L H SO as the color changed, and the
2
4
plate was analyzed using a multi-well spectrophotometer (SpectraMAX 190, Molecular Devices)
at 490 nm. The inhibition rate (%) was calculated using the following equation: [1-(A490/A490
control)] × 100%. The IC50 values were calculated from the inhibition curves in two separate
experiments.
5.4 Molecular modeling
Docking simulations were performed using the Molecular Operating Environment (MOE)
(
The Chemical Computing Group, Montreal, Canada). The crystal structure of c-Met crystal
2
3
26
structure(PDB code: 3LQ8) and VEGFR-2 crystal struture(PDB code: 3U6J) was downloaded
from the Protein Data Bank (PDB). Prior to ligand docking, hydrogens were added and the
12

crystallized ligand was removed. Subsequently, the structure was prepared with Protonate 3D, and
the active site was isolated using MOE Site Finder. The structures were placed in the site with the
Triangle Matcher method and then ranked with the London dG scoring function.For the energy
minimization in the pocket, MOE Force Field Refinement was used and ranked with the
GBVI/WSA dG scoring function.
Acknowledgments
This study was supported by Grants from the National Natural Science Foundation of China
(
Grants 81172932 and 81273376),the Natural Science Foundation of Jiangsu Province (Grant
BK2012356), and the Project Program of State Key Laboratory of Natural Medicines, China
Pharmaceutical University (GrantJKGZ201103).
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Design, synthesis and biological activity of 4-Aminopyrimidine-5-cabaldehyde oxime as dual
c-Met and VEGFR-2 inhibitor
1
Hao Qiang , DanDan Huang , Weijie Gu , Wei Shi , Qianqian Qiu , Yuxuan Dai , Wenlong
1,2
1
1
1
1
1
Huang , Hai Qian
,3*
1, 3*
1
Center of Drug Discovery, State Key Laboratory of Natural Medicines, China Pharmaceutical
University, 24 Tongjiaxiang, Nanjing 210009, PR China.
2
Department of Medicinal Chemistry, Nanjing Sanhome Pharmaceutical Co.,Ltd, Nanjing 210018,
PR China
3
Jiangsu Key Laboratory of Drug Discovery for Metabolic Disease, China Pharmaceutical
University, 24 Tongjiaxiang, Nanjing 210009, PR China
The cyclopropane-1, 1-dicarboxamide moiety might be critical for the potent dual inhibition of
c-Met and VEGFR-2. 4-Aminopyrimidine-5-cabaldehyde oxime as the scaffold of potent
VEGFR-2 inhibitor was combined with the cyclopropane-1,1-dicarboxamide moiety to obtain
potent c-Met and VEGFR-2 inhibitor.
15