New aminopyrimidine derivatives as inhibitors of the TAM family

Article abstract of DOI:10.1016/j.ejmech.2013.10.037

In this study, we describe the synthesis of new pyrimidine analogs of BMS-777607, a potent and selective inhibitor of Met kinase. Inhibition of Met and Axl remained high whereas inhibition of Tyro3 and Mer decreased to some extend. The preferential modera

Full text of DOI:10.1016/j.ejmech.2013.10.037

European Journal of Medicinal Chemistry 70 (2013) 789e801
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
New aminopyrimidine derivatives as inhibitors of the TAM family
Ténin Traoré a, Andrea Cavagnino b, Nicolas Saettel a, François Radvanyi d,
,
,c,
**
Sandrine Piguel a e, Isabelle Bernard-Pierrot d, Véronique Stoven b
,
Michel Legraverend a
,
*
a Institut Curie, CNRS, UMR 176, Bât. 110-112, Centre Universitaire, 91405 Orsay, France
b Mines ParisTech, CBIO ꢀ Centre for Computational Biology, 35 rue Saint-Honoré, F-77300 Fontainebleau, France
c Institut Curie, INSERM U900, Paris F-75248, France
d Institut Curie, CNRS, UMR 144, F-75248 Paris 05, France
e Univ Paris-Sud, Orsay F-91405, France
a r t i c l e i n f o
a b s t r a c t
Article history:
In this study, we describe the synthesis of new pyrimidine analogs of BMS-777607, a potent and selective
inhibitor of Met kinase. Inhibition of Met and Axl remained high whereas inhibition of Tyro3 and Mer
decreased to some extend. The preferential moderate inhibition of the non-phosphorylated form of Abl1
of some derivatives suggests that they behave as type II inhibitors. This hypothesis was confirmed by
docking studies into the structure of Met (3F82) and in a Tyro3 model where key interactions with the
hinge region, the DFG-out motif and the allosteric pocket explain this inhibition.
Received 19 June 2013
Received in revised form
10 October 2013
Accepted 12 October 2013
Available online 22 October 2013
Ó 2013 Elsevier Masson SAS. All rights reserved.
Keywords:
Pyrimidine synthesis
Met inhibitors
TAM inhibitors
Type 2 inhibitors
Molecular modeling
Docking
1. Introduction
non-specifically kill any actively dividing cells [7]. It has, however
to be noted that the 20 kinase inhibitor drugs known to date, target
Transmembrane receptor tyrosine kinases (RTKs) play funda-
mental roles in cellular processes, including proliferation, migra-
tion, metabolism, differentiation, and survival. The approval of
imatinib (Gleevec) for chronic myeloid leukemia (CML) [1] and
erlotinib (Tarceva) [2,3] for non-small cell lung cancer (NSCLC) has
provided proof-of-principle that small molecule kinase inhibitors
can be effective drugs. Currently, 20 kinase inhibitors are on the
market as drugs, mainly for the treatment of different types of
human tumors including pancreas, lung, kidney, breast, thyroid and
bone marrow, as well as CML, GIST (Gastro-intestinal stromal tu-
mors), and NSCLC [4e6]. These treatments have been designed to
selectively target oncogenic pathways in specific cancers, as
opposed to traditional highly cytotoxic chemotherapeutics which
a small number of kinases, although many other kinases are
involved in the processes leading to tumor cell proliferation and
survival and are presently under investigation. Among these new
therapeutic targets, Met and the three close proteins Tyro3, Axl and
Mer, defining the TAM subfamily of receptor protein kinases (the
four proteins forming the HGFR/TAM family) have appeared as
promising targets for different types of cancers [7e10]. In order to
target kinases, two main categories of kinase inhibitors have been
identified. Thus, type I inhibitors, form the largest of the two
classes, binding to the active site (the ATP-binding site) in the so-
called DFG-in active form of the enzyme, and type II inhibitors
which bind to a DFG-out conformation of the enzyme, where a
large hydrophobic allosteric pocket, adjacent to the active site,
becomes accessible [11,12]. The Met inhibitor BMS-777607 (com-
pound 1, Fig. 1), currently evaluated against solid tumors in a phase
II clinical trial [13,14], is a type II inhibitor that is also a potent in-
hibitor of the TAM family [15]. Conversely, various inhibitors of
Tyro3, Axl and Mer receptor protein kinases have been described
[7e9,16,17], but most of them also target Met, and, to date, no in-
hibitor of these kinases has been approved as an anticancer drug. In
* Corresponding author. Tel.: þ33 1 69 86 30 85.
** Corresponding author. Mines ParisTech, CBIO
ꢀ
Centre for Computational
Biology, 35 rue Saint-Honoré, F-77300 Fontainebleau, France. Tel.: þ33 1 56 24 69
28.
E-mail addresses: veronique.stoven@mines-paristech.fr (V. Stoven), michel.
legraverend@curie.fr (M. Legraverend).
0223-5234/$ e see front matter Ó 2013 Elsevier Masson SAS. All rights reserved.
http://dx.doi.org/10.1016/j.ejmech.2013.10.037

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T. Traoré et al. / European Journal of Medicinal Chemistry 70 (2013) 789e801
We also tested whether other substituents on the terminal phenyl
ring could modulate activity or selectivity (Fig. 1).
2. Results and discussion
2.1. Chemistry
Our initial SAR efforts began by investigating the importance of
the substituent on the terminal phenyl (p-fluorophenyl in 1, 2 and
3). For this purpose, aminophenoxypyrimidines 6 were prepared by
coupling 4,6-dichloropyrimidine 4 with 4-nitrophenol, followed by
reduction of the nitro group of intermediates 5 by Raney nickel
(Scheme 1).
Michael addition of the anilines 7aee to 3-carbomethoxy-2-
pyrone followed by intramolecular cyclization of the Michael
adduct provided the phenyl-2-pyridone ester 8aee [20], which
were hydrolyzed to pyridone carboxylic acids 9aee with LiOH
(Scheme 2).
Coupling of 3-fluoroaniline 6b with pyridone acid 9d in the
presence ofTBTU/DIPEAin DMFat 0ꢁC ledtothe6-chloropyrimidine
derivative10d (Scheme 3). However, the chlorineatomat 6could not
be substituted by para-methoxybenzylamine under various condi-
tions including in refluxing DME for 6 h. Interestingly, when the
coupling of carbocylic acids 9aee, with anilines 6 was carried out at
60 ꢁC, the O6-(benzotriazol-1-yl)pyrimidine derivatives 11aee were
isolated, resulting from the substitution of the chlorine atom by
HOBT which is formed during the amide coupling. As displacement
of O6-(benzotriazol-1-yl) from a purine (inosine) with a variety of
nucleophiles has been illustrated by Lakshman [21], we thought this
reaction would also be efficient in pyrimidines to introduce an
amino group. Indeed the O6-benzotriazol intermediates 11aee were
easilysubstituted with variousamines toprovidecompounds 12aee
and 14e15 in high yield. To the best of our knowledge, this is an
unprecedented example of the use of a O6-(benzotriazol-1-yl) as a
leaving group in pyrimidine series which can lead to 6-
aminopyrimidine derivatives by SNAr displacement. A similar
reaction was however reported previously [22], in the synthesis of
O2-pyrimidine ethers. Final hydrolysis of benzylamino derivatives
12aee in TFA led to amino derivatives 13aee.
In order to study the best position of the amino group (2 or 6) in
the pyrimidine series, isomers of compounds 12d, 13d, and 15 were
synthesized. For this purpose, aminophenoxypyrimidines 19a and
19b were prepared by coupling 2,4-dichloropyrimidine 16 with 2-
fluoro-4-nitrophenol, followed by smooth substitution of the 2-
chlorine atom by 4-methoxybenzylamine or methylamine,
respectively. Reduction of the nitro group of intermediates 18a and
18b was achieved by Zn/NH4Cl and led to 19a and 19b (Scheme 4).
Coupling of 19a and 19b with 9d was performed as for the synthesis
of 11aee (Scheme 3) and led to 20a and 20b, respectively while
treatment of 20a with TFA led to 2-aminopyrimidine derivative 21
(Scheme 5). Thus 20a, 21 and 20b (Scheme 5) are isomers of 12d,
13d and 15, respectively (Scheme 3).
Fig. 1. Probing the anti-TAM SAR around the pyrimidine scaffold (3); Met and TAM
family inhibitors BMS-777607 1 [15] and 2 [19].
this context of drug design, the selectivity profile is an important
issue, and it would be of high interest to design inhibitors selective
for any protein of the HGFR/TAM family. However, in view of the
high level of sequence similarity between Met and the enzymes of
the TAM family (44% identity and 60% similarity between Met and
Tyro3, and 46% identity and 64% similarity between Met and Mer),
the design of selective inhibitors within the HGFR/TAM family re-
mains a challenge. Because type II inhibitors are usually expected to
display better selectivity profiles, compound 1 appeared to us as a
possible starting molecule to search for more selective inhibitors
within the TAM family. Among them, we paid special attention to
Tyro3, a well validated drug target for bladder cancer [18], for
which no specific drug is available.
Based on biochemical and docking studies, we have previously
shown that the purine scaffold was also compatible with type II
inhibition against Met, Tyro3, Axl and Mer, as observed in com-
pound 2 (Fig. 1) [19]. We highlighted the necessity of the presence
of an ether linkage between the purine scaffold and the amido-
pyridone chain, since changing this linkage led to complete loss of
inhibitory activity against all considered kinases in the case of a
carbonecarbon bonding and to a reduction of inhibitory activity in
the case of an amino bonding, which was complete for Tyro3. The
presence of the amidopyridone group in this lateral chain was
found to be important (Fig. 1) [19].
In the present work, we explored the possibility to change the
purine scaffold to a pyrimidine in order to modulate the selectivity
profile of the inhibitor.
In addition, the effect of the presence and the position of the
NH2 was examined, since H-bond donor groups involved in pro-
teineligand interactions are present in similar positions in 1 and 2.
Compound 23 was then synthesized to be compared to 13d, in
order to evaluate the influence of the ortho fluoro atom on the
phenyl ether on the inhibitory activity. The same sequence used for
the synthesis of 13aee (Scheme 3) was used from 6-
chloropyrimidine 6a. As for 13aee, the amino derivative 23 was
obtained after treatment of the p-methoxybenzylamino precursor
with trifluoro acetic acid (Scheme 6).
2.2. Biological results
With the goal of targeting Tyro3, a first screening performed
against Tyro3 kinase domain using an enzyme-linked-
immunosorbent ELISA assay, showed that 10d, 11aee, 12d, 13b,
Scheme 1. Synthesis of intermediates 6. Reagents and conditions: (a) K2CO3, DMF,
80 ꢁC, 3 h, 87% (5a); 93% (5b); (b) Raney Ni, H2, MeOH, rt, 5 h, 47% (6a); 75% (6b).

T. Traoré et al. / European Journal of Medicinal Chemistry 70 (2013) 789e801
791
Scheme 2. Synthesis of the pyridone carboxylic acids 9aee. Reagents and conditions: (a) 1) DMF, 7 h, 0 ꢁC 2) EDCI, DMAP, 12 h, rt; yields: 8aee, 30, 34, 27, 52 and 21%, respectively
(b) LiOH, THF, 3 h, rt; yields: 9aee, 98, 96, 90, 96 and 92%, respectively.
Scheme 3. Coupling of carboxylic acids 9aee with aniline 6b and synthesis of 10d, 11, 12, 13, 14 and 15. Reagents and conditions: (a) TBTU, DIPEA, DMF, 0 ꢁC, 4 h; (b) TBTU, DIPEA,
DMF, 60 ꢁC, 4 h, yields: 11aee, 82, 82, 84, 88 and 84% respectively (c) p-MeO-benzylamine, Cs2CO3, DME, reflux, 6 h; yields: 12aee, 85, 90, 77, 94, 83% respectively (d) TFA, DCM,
reflux, 6 h, yields: 13aee, 74, 55, 83, 68 and 91% respectively; (e) Cs2CO3, DME, reflux, 2 h, amine: 4-(2-aminoethyl)morpholine, or methylamine, 76% (14), 55% (15). TBTU, O-
(Benzotriazol-1-yl)-N,N,N’,N’-tetramethyluronium tetrafluoroborate; DIPEA, diisopropylethylamine; DMF, dimethylformamide; DME, 1,2-dimethoxyethane; TFA, trifluoro acetic
acid; DCM, dichloromethane.
13e, 15, 20a and 20b were inactive when assayed up to 50
m
M (data
Scheme 3 for atom numbering). As already shown for 1, the presence
of an OEt substituent on the pyridone ring is not necessary for the
inhibition of Met [15]. In addition, the presence of an NH2 in posi-
tions 2 or 6 are both compatible with activity, and 21 and 13d have
very similar selectivity profiles against the kinases included in the
study. It should however be noted that BMS-777607 (1) was found to
inhibit the non-phosphorylated form of Abl1 (i.e. the DFG-out form),
not shown). The six active compounds against Tyro3 kinase were
then assayed against the two other proteins of the HGFR/TAM
family (Axl and Mer), Met, and a small panel of more distant
tyrosine kinases to characterize their selectivity profile (Table 1).
The pyridine ring in BMS-777607 (1) was replaced in 21 and 13d
by a pyrimidine ring substituted in position 2 and 6 respectively (see
Scheme 4. Synthesis of 4-amino-2-fluorophenoxy-pyrimidine intermediates 19a and 19b. Reagents and conditions: (a) K2CO3, DMF, 80 ꢁC, 3 h, yield: 58% (17); (b) K2CO3, DMF,
60 ꢁC, 30 min, p-MeOebenzyl-NH2, yield: 26% (18a) or MeNH2, yield: 16% (20); (c) Zn, NH4Cl, MeOH/THF 1:1, rt, 1 h, yields: 96% (19a), 99% (19b).

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T. Traoré et al. / European Journal of Medicinal Chemistry 70 (2013) 789e801
At this point, further analysis of Table 1 urged us to add that, in
the context of searching for Met inhibitors (which was not the main
goal here), some of the molecules synthesized in the present study
could be of interest. When compared to BMS-777607, all the mol-
ecules in Table 1 display similar inhibition activities against Met.
However, they are all less potent against Braf and Abl1. In addition,
they tend to be less active against Tyro3 and Mer, at least in the case
of 13a and 23. Overall, they present a better selectivity than the
reference molecule BMS-777607 currently under development as
an anticancer agent.
2.3. Molecular modeling
In order to provide structural interpretation for the biological
tests, we undertook molecular modeling studies. The synthesized
molecules derive from inhibitor 1, a type II inhibitor of Met, as
observed in the 3F82 PDB structure. As shown in Fig. 2 in the case of
1, type II inhibitors bind to the protein via several key interactions
including (i) at least one H-bond, but often two H-bonds, one as
donorand one as acceptor (with the backbone atoms of M1160 in the
case of 3F82), (ii) a hydrophobic interaction in the hinge region (with
the side chain of M1211 in the case of 3F82), (iii) interactions with
the DFG motif (a H-bond with the side chain of the D and a hydro-
phobic interaction with the F residue, corresponding to D1222 and
F1223 in 3F82), and hydrophobic interactions in the allosteric site.
Molecules that are unable to bind to the protein via these conserved
interactions are not expected to be type II inhibitors.
We docked the 14 synthesized molecules into the 3F82 structure
of Met, the only kinase from the HGFR/TAM family for which DFG-out
crystalstructureisavailable, and into a DFG-outmodelofstructure for
Tyro3 [19], and we analyzed their predicted binding modes.
When docked in Tyro3, all active molecules with an NH2 group
on the pyrimidine ring can form a double H-bond with M606, as
shown in Fig. 2 in the case of 13d. They also present all the other
interactions expected for type II inhibitors, as shown in Fig. 2
(poseview figure of 13d docked in 3F82 and in Tyro3). The same
result holds when these molecules are docked in 3F82 (as shown in
Table 1, all molecules active against Tyro3 are also active against
Met).
Scheme 5. Synthesis of the 2-amino derivatives 20 and 21. Reagents and conditions:
(a) TBTU, DIPEA, DMF, 0 ꢁC, 4 h, yields: 85% (20a), 40% (20b); (b) TFA, DCM, reflux, 12 h,
yield: 85% (21).
whereas 13d and 21 were less active against Abl1. Interestingly, they
exhibited more activity against the unphosphorylated form of Abl1
than the phosphorylated Abl1, which shows that they behave as
type II inhibitors, at least against Abl1. Similarly, at high concen-
tration of inhibitors, 13d and 21 were less active than BMS-777607
against Braf and EGFR. These two observations indicate that sub-
stitution of the pyridine ring by a pyrimidine ring seems to increase
the specificity towards the four proteins of the HGFR/TAM family.
Such improvement in selectivity is also shared by compound 2, in
which the pyridine ring of BMS-777607 was replaced by a purine
ring [19].
23 only differs from 13d by the absence of a fluorine atom on the
phenyl group born by the pyrimidine scaffold. These two molecules
present very similar activities against Met and the TAM family,
which shows that the presence of this fluorine atom is not crucial
for the inhibitory activity. Substitution of the fluorine atom of the
terminal phenyl by a larger O-Met group moiety, as in 13e, leads to
a loss of activity. Similar results have already been observed in the
case of compound 2, where replacement of F atom by a CF3 group
suppressed inhibition against Tyro3 [19].
Adding a second substituent of moderate size in ortho of the
terminal phenyl (13a, substitution with F or 13c, substitution with
CH3) is compatible with inhibition activity, although with a loss in
the case of Tyro3 and Mer. These substitutions, however did not
affect potency against Axl and Met, whereas substitution in meta of
the terminal phenyl (13b), led to complete loss of inhibitory activity
against Tyro3.
All compounds bearing no H-bond donor group on the pyrimi-
dine ring were found to be inactive against Tyro3, namely com-
pounds 11a and 10d. Most of the compounds bearing a substituted
eNH group on the pyrimidine ring as H-bond donors (instead of
NH2) were also found to be inactive, namely compounds 12d, 15,
20a, and 20b, molecule 14 being the only exception to this
observation.
On the contrary, inactive molecules against Tyro3 bearing no H-
bond donor group substituted on the pyrimidine ring, namely
compounds 11a and 10d, were unable to form an H-bond with
M606 when docked in the model of Tyro3 (as shown in Fig. 2 in the
case of 10d), or with M1160 when docked in 3F82.
The same result was observed for the inactive molecules bearing
a substituted eNH group as H-bond donors on the pyrimidine ring,
namely compounds 12d, 15, 20a, and 20b. Indeed, a substituent on
the eNH group leads to displacements of the pyrimidine group
within the ATP-binding site in order to avoid steric hindrance,
preventing the formation of the two conserved H-bonds.
Substitution of the para-F atom on the terminal phenyl by a
larger group such as O-Met, as in the 13e molecule, leads to a
displacement of the docking pose with respect to the reference
Scheme 6. Synthesis of 6-aminopyrimidine derivative 23. Reagents and conditions: (a) TBTU, DIPEA, DMF, 0 ꢁC, 4 h, then 60 ꢁC, 12 h, yield: 72% (22); (b) 1) p-MeOebenzyl-NH2,
DME, reflux, 6 h 2) TFA, DCM, reflux, 12 h, yield for two steps: 69% (23).

Table 1
The percent inhibition refers to the percentage of kinase captured from the solid support that has been displaced by the inhibitor [23]; Concentrations tested: 0.5, 1 and 75 mM; nd, not determined.
% Inhibition at various concentrations in mM
0.5
1
75
0.5
Axl
1
75
0.5
1
75
95
0.5
1
75
96
0.5
1
75
0.5
1
75
99
0.5
1
0
75
95
0.5
1
75
78
0.5
1
0
75
0
Protein
Tyro3
Mer
Met
Abl1-non phosphor.
Abl1-phosphor.
Braf
EGFR
FGFR3
96
99
92
99
99
99
98
98
96
95
96
98
100
37
35
0
0
1
0
73
94
100
98
100
98
77
93
nd
93
98
97
95
17
32
66
0
0
20
0
0
43
0
9
0
0
0
16
27
48
77
50
63
65
94
63
100
100
100
75
74
93
97
95
93
98
100
100
100
100
50
53
89
90
71
65
93
93
87
99
83
49
91
99
98
98
99
99
98
99
25
10
23
12
37
27
41
12
85
98
94
71
8
0
0
1
4
0
0
0
21
47
39
13
0
0
4
6
0
0
0
0
11
29
21
11
7
4
5
9
8
0
17
19
14
8
0
0
0
0
0
0
0
0
9
6
100
98
100
99
10
16
99
8
0
(continued on next page)

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T. Traoré et al. / European Journal of Medicinal Chemistry 70 (2013) 789e801
molecule 1, which might explain reduced hydrophobic interactions
in the allosteric pocket, and loss of inhibition activity.
Docking studies were however unable to explain why the di-
fluoro derivative (in meta and para) was not compatible with in-
hibition of Tyro3 as in 13b, whereas the ortho-para-difluoro-
substituted derivative 13a exhibited only reduced potency.
3. Conclusion
The aim of this work was to modify the selectivity profile of the
Met inhibitor 1, and to increase the inhibitory potency in favor of
Tyro3. For this purpose, pyrimidine analogs of 1 have been syn-
thesized and tested against Tyro3 and a panel of other kinases. The
synthesis of derivatives bearing an NH2 group at position 6 of the
pyrimidine ring was obtained thanks to the unanticipated substi-
tution of the chlorine atom at position 6 of various pyrimidines by
an oxybenzotriazol during the peptide coupling at 60 ꢁC, which was
beneficial as this chlorine atom could not be substituted with p-
methoxybenzylamine under reflux of DME (Scheme 3). On the
contrary, SNAr displacement of the O6-benzotriazol could be ach-
ieved under mild conditions, with several amines, and in particular,
with methoxybenzylamine which gave an easy access to various
free 6-aminopyrimidine derivatives 13aee after acid hydrolysis.
All derivatives were assayed against Tyro3 tyrosine kinase and the
most active inhibitors were evaluated against the HGFR/TAM family
and a panel of more distant kinases. We identified 6 pyrimidine in-
hibitors, namely 13a, 13c, 13d, 14, 21, and 23, with a slight decrease in
potency against Tyro3 and Mer. Compounds 13c, 13d, 14 and 21 were
the most potent inhibitors of Axl and Met, and remained not active
against more distant kinases such as Braf, EGFR and FGFR3. They are
therefore selective. Furthermore, the weak inhibition of Abl1 in phos-
phorylated and unphosphorylated forms by the pyrimidine inhibitors
13aed,14, 21 and 23 deserves to be noted, as compared to 1 which is a
strong inhibitor of Abl1, at least in its unphosphorylated form.
The presence of an exocyclic NH2 on either side of the pyrimi-
dine N-1 is crucial for maintaining the inhibitory activity, since
replacement of such an NH2 by a chlorine atom (in 10d) canceled
the inhibition of Tyro3. Docking studies suggested that this NH2
group is required to form two H-bonds in the ATP-binding pocket,
as observed with known type II inhibitors.
To improve the selectivity for Tyro3 within this family, com-
parison of its ligand-binding pocket with those of Axl, Mer and Met
reveals the presence of a unique subpocket near the ATP binding
site due to the presence of Ala 581 in human Tyro3, a residue
replaced by amino acids with larger side-chains at this position in
Axl, Mer and Met. Our current efforts are aimed at synthesizing
new type II inhibitors that would occupy this Ala 581 pocket, to
improved selectivity for Tyro3, as shown by Powell in murine Sky
for type I inhibitors [25].
4. Experimental section
4.1. Chemistry
NMR spectra were recorded at 300 MHz (1H NMR) or 75.3 MHz
(
13C NMR) with CDCl3, CD2Cl2 or DMSO-d6 as solvents and tetra-
methylsilane as internal standard on a Bruker AC300 spectrometer.
Chemical shifts are expressed in ppm ( ) downfield from TMS. J
d
values are expressed in Hertz (Hz). The following abbreviations are
used for the multiplicities: s, singlet; d, doublet; t, triplet; m,
multiplet; b, broad. MS spectra were recorded on a Waters ZQ2000
mass spectrometer with direct injection. HRMS spectra were per-
formed by the mass spectrometry service Imagif at the ICSN (Gif-
sur-Yvette). Anilines are commercially available. Abbreviations
used for reagents are given in the legend of Scheme 3.

T. Traoré et al. / European Journal of Medicinal Chemistry 70 (2013) 789e801
795
Fig. 2. Proteineligand interactions between (a) 1 in Met (PDB code 3F82), (b) 13d docked in Tyro3, (c) 10d docked in Tyro3. Figures were generated using Poseview [24].
4.1.1. General procedure for the synthesis of 4-chloro-6-(4-
nitrophenoxy)pyrimidine (5a) and 4-chloro-6-(2-fluoro-4-
nitrophenoxy)pyrimidine (5b)
at room temperature for 3 h. The catalyst was then filtered over a
pad of celite, washed with EtOAc and DCM and the filtrate was
evaporated. The brown residue was extracted with EtOAc. The
organic layer was washed with water and brine; dried over MgSO4
and filtered. The solvent was evaporated to give the crude product
as a brown solid. This residue was purified by flash chromatography
using Cyclohexane/Ethyl acetate 65:35 as eluent to afford the
desired product as a solid.
In
a dry sealed tube under argon were placed, 4,6-
dichloropyrimidine (1 g, 6.7 mmol), 4-nitrophenol (1.1 g, 7.9 mmol),
or 2-fluoro-4-nitrophenol (1.24 g, 7.9 mmol), (potassium carbonate
(1.3 g, 9.3 mmol) in DMF (13 mL) and the mixture was heated at 80 ꢁC
for 3 h. The mixture was then cooled at room temperature and the
solvent evaporated under high vacuum. The residue was extracted
with EtOAc, washed with a saturated solution of NaHCO3, water and
brine. The organic layers were dried over MgSO4, filtered and the
solvent was evaporated to give the crude product as a yellow solid.
The residue was purified by flash chromatography using Cyclo-
hexane/DCM35: 65 as eluent to afford the desired product which was
used in the next step without any further purification.
4.1.2.1. 4-((6-Chloropyrimidin-4-yl)oxy) aniline (6a). A white solid
was obtained in 47% yield. 1H NMR (DMSO-d6, 300 MHz)
d
(ppm):
8.62 (s, 1H), 7.13 (s, 1H), 6.99e6.78 (m, 2H), 6.70e6.49 (m, 2H), 5.13
(s, 2H). 13C NMR (DMSO-d6, 75 MHz)
d (ppm): 170.8, 160.6, 158.6,
146.8, 141.7, 121.6, 114.3, 107.2. MS (EI) ESþ: 222 ([Mþ þ H], 100).
4.1.2.2. 4-((6-Chloropyrimidin-4-yl)oxy)-3-fluoroaniline
(6b).
4.1.1.1. 4-Chloro-6-(4-nitrophenoxy)pyrimidine (5a). Light yellow
This compound was obtained with 75% yield according to the
solid in 87% yield. 1H NMR (DMSO-d6, 300 MHz)
d
(ppm): 8.70 (s,
general procedure. 1H NMR (DMSO-d6, 300 MHz)
d (ppm): 8.64 (s,
1H), 8.36 (d, J ¼ 12 Hz, 2H), 7.6 (s, 1H), 7.57 (d, J ¼ 12 Hz, 2H). 13
C
1H), 7.40 (s, 1H), 6.97 (t, J ¼ 9 Hz, 1H), 6.49e6.44 (dd, J ¼ 9 Hz,
NMR (DMSO-d6, 75 MHz)
d (ppm): 169, 161.2, 158.4, 156.8, 144.9,
J ¼ 6 Hz, 1H), 6.40e6.36 (m, 1H), 5.44 (s, 2H). 13C NMR (DMSO-d6,
125.6, 122.8, 108.9. MS (EI) ESþ: 252 ([Mþ þ H], 100).
75 MHz) d (ppm): 169.9, 160.8, 158.8, 155.5, 152.3, 148.7, 148.5, 128,
127.8, 123.7, 123.6, 109.6, 109.5, 107.3, 101.1, 100.8 MS (EI) ESþ: 240
4.1.1.2. 4-Chloro-6-(2-fluoro-4-nitrophenoxy)pyrimidine
(5b).
([Mþ þ H], 100).
This compound was obtained with 93% yield according to the general
method. 1H NMR (DMSO-d6, 300 MHz)
d
(ppm): 8.70 (d, J ¼ 3 Hz,1H),
4.1.3. General procedure A for the synthesis of substituted pyridones
(8aee)
8.43e8.39(dd, J ¼ 6 Hz, J ¼ 3 Hz,1H), 8.24e8.20(m,1H), 7.82e7.77 (m,
1H), 7.75 (d, J ¼ 3 Hz,1H).13C NMR (DMSO-d6, 75MHz)
d
(ppm): 169.5,
In a dry flaskunderargon, asolutionofmethyl-2-oxo-2H-pyran-3-
carboxylate (1.5 g, 9.7 mmol) in DMF (10 mL) was placed at 0 ꢁC and
thecorrespondingsubstituted aniline(10mmol)was added dropwise
tothemixtureandstirred7hatthistemperature.ThenEDCI(N-Ethyl-
N'-(3-dimethylaminopropyl)carbodiimide) (2.4 g, 12 mmol) and
DMAP (4-Dimethylaminopyridine) (0.3 g, 2.4 mmol) were added to
the mixture and stirred at room temperature for 12 h. The solvent was
removed under high vacuum and the brown oil was extracted with
EtOAc, washed with water, brine, dried over MgSO4 and filtered. The
solventwasevaporated togivethecrudeproductasa brownoilwhich
was purified by flash chromatography to afford the desired product.
168.2, 161.5, 158.4, 154.7, 151.4, 145.7, 145.6, 144.2, 144.1, 124.9, 121.1,
121, 113.3, 113, 108.3. MS (EI) ESþ: 270 ([Mþ þ H], 100).
4.1.2. General procedure for the synthesis of 4-((6-chloropyrimidin-
4-yl)oxy) aniline (6a), and 4-((6-chloropyrimidin-4-yl)oxy)-3-
fluoroaniline (6b)
A solution of 4-chloro-6-(4-nitrophenoxy)pyrimidine 5a (0.5 g,
1.98 mmol) or 4-chloro-6-(2-fluoro-4-nitrophenoxy)pyrimidine 5b
(0.5 g, 1.85 mmol) in methanol (48 mL) was treated with Ni Raney
(1.2 g aqueous slurry) and the mixture was stirred under hydrogen

796
T. Traoré et al. / European Journal of Medicinal Chemistry 70 (2013) 789e801
4.1.3.1. Methyl-1-(2,4-difluorophenyl)-2-oxo-1,2-dihydropyridine-3-
carboxylate (8a). From 2,4-difluoroaniline (7a). Flash chromatog-
raphy using DCM/Ethyl acetate 95:5 as eluent affords the desired
yield. 1H NMR (DMSO-d6, 300 MHz)
d (ppm): 13.8 (s, 1H), 8.51
(dd, J ¼ 3 Hz, J ¼ 6 Hz, 1H), 8.24 (dd, J ¼ 3 Hz, J ¼ 6 Hz, 1H), 7.79e
7.71 (m, 1H), 7.65e7.58 (m, 1H), 7.36e7.30 (m, 1H), 6.83 (t,
product with 30% yield. 1H NMR (DMSO-d6, 300 MHz)
d
(ppm): 8.18
J ¼ 6 Hz, 1H). 13C NMR (DMSO-d6, 75 MHz)
d (ppm): 164.3, 164.1,
(dd, J ¼ 3 Hz, J ¼ 6 Hz,1H), 7.98 (dd, J ¼ 3 Hz, J ¼ 6 Hz,1H), 7.64e7.54
162.7, 160.9, 160.8, 158.4, 154.2, 155, 154.9, 146.8, 145.2, 130.6,
130.4, 123.3, 123.1, 117.6, 112.6, 112.2, 108.3, 105.4, 105.1, 104.8.
MS (EI) ESþ: 274 ([Mþ þ Na], 100). MS (EI) ESꢀ: 250 ([Mꢀ ꢀ H],
90).
(m, 2H), 7.31e7.25 (m, 1H), 6.47 (t, J ¼ 6 Hz, 1H), 3.75 (s, 3H). 13
C
NMR (DMSO-d6, 75 MHz) d (ppm): d 164.7, 163.8, 163.7, 160.5, 158.7,
158.5, 157.4, 155.3, 155.1, 145.4, 144.4, 130.6, 130.5, 124.6, 124.5,
124.4, 124.3, 120.4, 112.4, 112.3, 112.1, 112.0, 105.2, 104.9, 104.8,
104.5, 51.8. MS (EI) ESþ: 266 ([Mþ þ H], 100), 288 ([Mþ þ Na], 70).
4.1.4.2. 1-(3,4-Difluorophenyl)-2-oxo-1,2-dihydropyridine-3-
carboxylic acid (9b). The desired product was obtained with 96%
4.1.3.2. Methyl-1-(3,4-difluorophenyl)-2-oxo-1,2-dihydropyridine-3-
carboxylate (8b). From 3,4-difluoroaniline (7b) Flash chromatog-
raphy using DCM/Ethyl acetate 95:5 as eluent affords the desired
yield. 1H NMR (DMSO-d6, 300 MHz)
d (ppm): 14.07 (s, 1H), 8.50 (dd,
J ¼ 3 Hz, J ¼ 6 Hz, 1H), 8.22 (dd, J ¼ 3 Hz, J ¼ 6 Hz, 1H), 7.88e7.81 (m,
1H), 7.70e7.63 (m,1H), 7.49 (t, J ¼ 3 Hz,1H), 6.81 (t, J ¼ 6 Hz,1H). 13
C
product with 34% yield. 1H NMR (DMSO-d6, 300 MHz)
d
(ppm): 8.14
NMR (DMSO-d6, 75 MHz) d (ppm): 164.4, 163.4, 151.4, 151.3, 150.2,
(dd, J ¼ 3 Hz, J ¼ 6 Hz,1H), 7.97 (dd, J ¼ 3 Hz, J ¼ 6 Hz, 1H), 7.76e7.57
148.1, 148, 147.4, 146.4, 144.9, 135.5, 135.4, 124.4, 118.1, 117.8, 117.5,
117, 108.2. MS (EI) ESþ: 274 ([Mþ þ Na], 100). MS (EI) ESꢀ: 250
([Mꢀ ꢀ H], 90).
(m, 2H), 7.35e7.21 (m, 1H), 6.44 (t, J ¼ 6 Hz, 1H), 3.74 (s, 3H). 13
C
NMR (DMSO-d6, 75 MHz)
d (ppm): 164.8, 157.9, 150.9, 150.4, 147.7,
145, 143.9, 136.9, 136.8, 136.7, 124.5, 124.4, 124.3, 120.6, 117.8, 117.6,
117.3, 117.1, 104.6, 51.8. MS (EI) ESþ: 266 ([Mþ þ H], 10); 288
([Mþ þ Na], 100).
4.1.4.3. 1-(4-Fluoro2-methylphenyl)-2-oxo-1,2-dihydropyridine-3-
carboxylic acid (9c). The desired product was obtained with 90%
yield. 1H NMR (DMSO-d6, 300 MHz)
d (ppm): 14.16 (s, 1H), 8.53 (dd,
4.1.3.3. Methyl-1-(4-fluoro-2-methylphenyl)-2-oxo-1,2-
dihydropyridine-3-carboxylate (8c). From 4-fluoro-2-methylaniline
(7c). Flash chromatography using 2%e5% Ethanol/DCM as eluent
affords the desired product with 27% yield. 1H NMR (DMSO-d6,
J ¼ 3 Hz, J ¼ 6 Hz, 1H), 8.15 (dd, J ¼ 3 Hz, J ¼ 6 Hz, 1H), 7.50e7.46 (m,
1H), 7.37e7.33 (dd, J ¼ 3 Hz, J ¼ 9 Hz, 1H), 7.25e7.21 (td, 1H), 6.83 (t,
J ¼ 6 Hz, 1H). 13C NMR (DMSO-d6, 75 MHz)
d (ppm): 164.5, 163.6,
163.3, 160.4, 146.4, 145.1, 137.4, 137.2, 134.8, 134.7, 129.4,129.3, 117.5,
117.2, 114, 113.7, 108.5, 16.9. MS (EI) ESꢀ: 246 ([Mꢀ ꢀ H], 90), 202
([Mꢀ ꢀ CO2H], 70).
300 MHz)
J ¼ 6 Hz, 1H), 7.36e7.27 (m, 2H), 7.21e7.16 (m, 1H), 6.44 (t, J ¼ 6 Hz,
1H), 3.74 (s, 3H). 13C NMR (DMSO-d6, 75 MHz)
(ppm): 164.9,163.3,
d
(ppm): 8.17 (dd, J ¼ 3 Hz, J ¼ 6 Hz,1H), 7.86 (dd, J ¼ 3 Hz,
d
160, 157.7, 145, 144.3, 137.5, 137.4, 136, 129.5, 129.3, 120, 117.2, 117,
4.1.4.4. 1-(4-Fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylic
113.8, 113.5, 104.6, 51.8, 17.1. MS (EI) ESþ: 284 ([Mþ þ Na], 100).
acid (9d). The desired product was obtained with 96% yield. 1H
NMR (DMSO-d6, 300 MHz)
d
(ppm): 14.23 (s, 1H), 8.49 (d, J ¼ 6 Hz,
4.1.3.4. Methyl-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-
carboxylate (8d). From 4-fluoroaniline (7d). Flash chromatography
using 0.5% Ethanol/DCM as eluent affords the desired product with
1H), 8.21 (d, J ¼ 6 Hz, 1H), 7.62e7.59 (m, 2H), 7.44e7.39 (td, 2H),
6.81 (t, J ¼ 9 Hz, 1H). 13C NMR (DMSO-d6, 75 MHz)
d (ppm): 164.6,
163.7, 160.4, 146.3, 145.2, 135.4, 129.2, 129.0, 117.4, 116.3, 116.0,
108.3. MS (EI) ESþ: 256 ([Mþ þ Na], 100). MS (EI) ESꢀ: 232 ([M ꢀ H],
100).
52% yield. 1H NMR (DMSO-d6, 300 MHz)
d
(ppm): 8.25 (dd, J ¼ 9 Hz,
1H), 7.57 (dd, J ¼ 6 Hz, 1H), 7.33e7.38 (m, 2H), 7.17 (t, J ¼ 9 Hz, 2H),
6.34 (t, J ¼ 6 Hz, 1H), 3.90 (s, 3H). 13C NMR (DMSO-d6, 75 MHz)
d
(ppm): 164.1, 160.8, 145.0, 143.3, 136.1, 128.6, 128.5, 122.0, 116.5,
4.1.4.5. 1-(2-Fluoro-4-methoxyphenyl)-2-oxo-1,2-dihydropyridine-
116.2, 105.5, 51.8. MS (EI) ESþ: 270 ([Mþ þ Na], 100).
3-carboxylic acid (9e). The desired product was obtained with 92%
yield. 1H NMR (DMSO-d6, 300 MHz)
d (ppm): 13.97 (s, 1H), 8.51 (dd,
4.1.3.5. Methyl-2-oxo-1-(2-fluoro-4-methoxyphenyl)-1,2-
J ¼ 3 Hz, J ¼ 6 Hz, 1H), 8.21 (dd, J ¼ 3 Hz, J ¼ 6 Hz, 1H), 7.58 (t,
J ¼ 9 Hz, 1H), 7.16 (dd, J ¼ 3 Hz, J ¼ 9 Hz, 1H), 6.98 (dd, J ¼ 3 Hz,
J ¼ 9 Hz, 1H), 6.81 (t, J ¼ 6 Hz, 1H), 3.84 (s, 1H). 13C NMR (DMSO-d6,
75 MHz) d (ppm): 164.4, 163.1, 161.4, 161.3, 158.5, 155.2, 146.6, 145.7,
129.4, 119.2, 119, 117.3, 110.9, 108.3, 102.4, 102.1, 56.1. MS (EI) ESþ:
286 ([Mþ þ Na], 100).
dihydropyridine-3-carboxylate
(8e). From
2-fluoro-4-
methoxyaniline (7e). Flash chromatography using DCM/Ethyl ace-
tate 95:5 as eluent affords the desired product with 21% yield. 1H
NMR (DMSO-d6, 300 MHz)
d
(ppm): 8.16 (dd, J1 ¼ 3 Hz, J2 ¼ 6 Hz,
1H), 7.95 (dd, J ¼ 3 Hz, J ¼ 6 Hz, 1H), 7.46 (t, J ¼ 9 Hz, 1H), 7.10 (dd,
J1 ¼ 3 Hz, J2 ¼ 9 Hz, 1H), 6.94 (dd, J ¼ 3 Hz, J ¼ 6 Hz, 1H), 6.44 (t,
J ¼ 6 Hz,1H), 3.83 (s, 3H), 3.75 (s, 3H). 13C NMR (DMSO-d6, 75 MHz):
4.1.5. General procedure C for the synthesis of N-(4-(6-
chloropyrimidin-4-yloxy)-3-fluorophenyl)-1-(4-fluorophenyl)-2-
oxo-1,2-dihydropyridine-3-carboxamide (10d) and N-(4-((6-((1H-
benzo[d][1,2,3]triazol-1-yl)oxy)pyrimidin-4-yl)oxy)-3-
fluorophenyl)-2-oxo-1-phenyl-1,2-dihydropyridine-3-carboxamide
(11aee)
In a dry flask under argon, were placed the corresponding car-
boxylic acid (0.68 mmol), TBTU (0.81 mmol) in DMF (1 mL) and the
solution was cool down at 0 ꢁC. Then DIPEA (1.8 mmol) was added
and the mixture was stirred during 15 min. The amino-pyrimidine
6b (0.62 mmol) was added and the mixture stirred 4 h at 0 ꢁC for
compound 10d, then 12 h at 60 ꢁC for compounds 11aee. The
solvent was removed under high vacuum and the brown oil was
extracted with EtOAc, washed with water, brine, dried over MgSO4
and filtered. The solvent was evaporated to give the crude product
as a brown solid which was purified by flash chromatography to
afford the desired product.
d
(ppm): 164.8, 160.8, 160.7, 158.8, 157.6, 155.5, 145.1, 144.7, 129.6,
129.5, 120.6, 120.5, 120.4, 110.6, 110.5, 104.6, 102.3, 102, 55.9, 51.8.
MS (EI) ESþ: 278 ([Mþ þ H], 80); 300 ([Mþ þ Na], 30).
4.1.4. General procedure B for the synthesis of substituted pyridone
carboxylic acids (9aee)
A solution of the corresponding substituted pyridone ester
(1.4 g, mmol) in THF (45 mL) at room temperature was treated with
an aqueous solution of LiOH 1 M (10 mL) during 3 h. The solvent
was evaporated; the residue was extracted with EtOAc and the pH
was adjusted to 2. The organic layer was dried over MgSO4, filtered
and the solvent was removed to give the crude product as a yellow
powder, which will be used without any further purification.
4.1.4.1. 1-(2,4-Difluorophenyl)-2-oxo-1,2-dihydropyridine-3-
carboxylic acid (9a). The desired product was obtained with 98%

T. Traoré et al. / European Journal of Medicinal Chemistry 70 (2013) 789e801
797
4.1.5.1. N-(4-(6-Chloropyrimidin-4-yloxy)-3-fluorophenyl)-1-(4-
fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide (10d).
1H NMR (CD2Cl2, 300 MHz)
4.1.5.6. N-(4-((6-((1H-Benzo[d][1,2,3]triazol-1-yl)oxy)pyrimidin-4-
yl)oxy)-3-fluorophenyl)-1-(2-fluoro-4-methoxyphenyl)-2-oxo-1,2-
dihydropyridine-3-carboxamide (11e). Flash chromatography using
DCM/Ethyl acetate 98:2 as eluent affords the desired product with
d
(ppm): 12.09 (s, 1H), 8.74 (dd, J ¼ 6 Hz,
J ¼ 2 Hz, 1H), 8.56 (s, 1H), 8.0 (dd, J ¼ 12 Hz, J ¼ 2 Hz, 1H), 7.69 (dd,
J ¼ 6 Hz, J ¼ 2 Hz, 1H), 7.46e7.41 (m, 2H), 7.32e7.26 (m, 3H), 7.18 (t,
J ¼ 8 Hz, 1H), 7.06 (s, 1H), 6.66e6.61 (t, J ¼ 6 Hz, 1H). 13C NMR
84% yield. 1H NMR (CDCl3, 300 MHz)
d (ppm): 11.98 (s, 1H), 8.76 (dd,
J1 ¼ 3 Hz, J2 ¼ 9 Hz, 1H), 8.34 (s, 1H), 8.14 (d, J ¼ 9 Hz, 1H), 7.98 (dd,
J ¼ 3 Hz, J ¼ 12 Hz, 1H), 7.61e7.30 (m, 6H), 7.19 (t, J ¼ 9 Hz, 1H), 6.88
(m, 2H), 6.66 (s, 1H), 6.64 (t, J ¼ 9 Hz, 1H), 3.88 (s, 3H). 13C NMR
(CDCl3, 75 MHz)
d (ppm): 169.6, 164.4, 162.4, 161.9, 161.4, 161.1,
158.4, 155.6, 152.3, 145.2, 141.7, 137.6, 137.5, 135.8, 135.7, 135.1, 134.9,
128.4, 128.3, 123.3, 121.9, 117, 116.7, 116.2, 116.1, 109.5, 109.2, 107.6,
107.3. MS (EI) ESþ: 477 ([Mþ þ Na], 100). HRMS-ESI (m/z) calcd for
(CDCl3, 75 MHz)
d (ppm): 171.2, 162.2, 161.9, 161.7, 161.5, 158.3,
155.8, 152.3, 145.3, 143.3, 142.5, 137.7, 135.2, 135, 129, 128.7, 128.5,
125, 123.3, 121.8, 120.5, 120, 119.8, 110.8, 110.7, 108.6, 107.2, 102.7,
90.3, 60.4, 60.3, 56.2, 14.2. MS (EI) ESþ: 585 ([Mþ þ H], 100).
C
22H14ClF2N4O3 (M þ Hþ) 455.0720, found: 455.0716.
4.1.5.2. N-(4-((6-((1H-Benzo[d][1,2,3]triazol-1-yl)oxy)pyrimidin-4-
yl)oxy)-3-fluorophenyl)-1-(2,4-difluorophenyl)-2-oxo-1,2-
dihydropyridine-3-carboxamide (11a). Flash chromatography using
1%e5% Ethyl acetate/DCM as eluent affords the desired product
4.1.6. General procedure D for the synthesis of 1-(substituted
phenyl)-N-(3-fluoro-4-(6-(4-methoxybenzylamino)pyrimidin-4-
yloxy)phenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide (12aee)
In a dry flask under argon, a solution of the corresponding
benzotriazol 11 (0.36 mmol), 4-methoxy-benzylamine (0.43 mmol)
and cesium carbonate (0.72 mmol) in DME (6 mL) was heated at
reflux during 6 h. The solvent was removed, the residue was
extracted with EtOAc, washed with water, brine, dried over MgSO4
and filtered. The solvent was evaporated to give the crude product
which was purified by flash chromatography to afford the desired
product.
with 82% yield. 1H NMR (CDCl3, 300 MHz)
d (ppm): 11.86 (s, 1H),
8.78 (dd, J ¼ 3 Hz, J ¼ 6 Hz, 1H), 8.35 (s, 1H), 8.14 (d, J ¼ 6 Hz, 1H),
7.98 (dd, J ¼ 3 Hz, J ¼ 12 Hz, 1H), 7.61e7.36 (m, 7H), 7.20e7.08 (m,
3H), 6.67e6.63 (m, 2H). 13C NMR (CDCl3, 75 MHz)
d (ppm): 171.3,
171.2, 161.9, 161.2, 158.3, 155.6, 152.3, 145.6, 143.4, 141.9, 137.6, 137.4,
135.3, 129, 128.5, 125, 123.3, 122, 120.6, 116.3, 116.2, 109.5, 109.2,
108.6,107.5, 90.3, 60.4, 21,14.2. MS (EI) ESþ: 594 ([Mþ þ Na],10). MS
(EI) ESꢀ: 570 ([Mꢀ ꢀ H], 100); 616 ([Mþ þ CO2H], 30).
4.1.5.3. N-(4-((6-((1H-Benzo[d][1,2,3]triazol-1-yl)oxy)pyrimidin-4-
yl)oxy)-3-fluorophenyl)-1-(3,4-difluorophenyl)-2-oxo-1,2-
dihydropyridine-3-carboxamide (11b). Flash chromatography using
1%e5% Ethyl acetate/DCM as eluent affords the desired product
4 .1. 6 .1. 1 - ( 2 , 4 - D i fl u o ro p h e n yl ) - N - ( 3 - fl u o r o - 4 - ( 6 - ( 4 -
methoxybenzylamino)pyrimidin-4-yloxy)phenyl)-2-oxo-1,2-
dihydropyridine-3-carboxamide (12a). Flash chromatography using
DCM/Ethyl acetate 70:30 as eluent affords the desired product with
with 82% yield. 1H NMR (CDCl3, 300 MHz)
d
(ppm): 11.91 (s, 1H),
85% yield. 1H NMR (DMSO-d6, 300 MHz)
d (ppm): 11.85 (s, 1H), 8.63
8.77 (dd, J ¼ 3 Hz, J ¼ 9 Hz, 1H), 8.35 (s, 1H), 8.14 (d, J ¼ 9 Hz, 1H),
7.97 (dd, J ¼ 3 Hz, J ¼ 9 Hz, 1H), 7.63e7.57 (m, 2H), 7.54e7.43 (m,
2H), 7.41e7.30 (m, 2H), 7.24e7.15 (m, 3H). 13C NMR (CDCl3, 75 MHz)
(dd, J ¼ 6 Hz, J ¼ 2 Hz, 1H), 8.17e8.11 (m, 2H), 7.93e7.72 (m, 3H),
7.65e7.58 (m, 1H), 7.42e7.17 (m, 5H), 6.90 (d, J ¼ 9 Hz, 2H), 6.79 (t,
J ¼ 6 Hz, 1H), 5.95 (s, 1H), 4.43 (s, 2H), 3.72 (s, 3H). 13C NMR (DMSO-
d
(ppm): 171.3, 171.2, 162.2, 161.2, 158.3, 155.6, 152.3, 152.1, 149.2,
d6, 75 MHz) d (ppm): 168.1, 164.5, 164, 162.8, 161.2, 160.8, 158.6,
148.7, 145.4, 143.4, 141.3, 137.6, 137.4, 135.7, 135.3, 135.1, 129, 128.5,
125, 123.4, 122.1, 120.6, 118.5, 116.2, 109.5, 109.2, 108.6, 107.5, 90.3.
MS (EI) ESꢀ: 570 ([Mꢀ ꢀ H], 100); 616 ([Mþ þ CO2H], 30).
158.2, 155.2, 155, 152, 145.5, 144.2, 136.4, 136.3, 135.4, 135.3, 130.7,
130.6, 128.5, 124.3, 124, 123.8, 123.1, 120.1, 116.1, 113.7, 112.5, 112.2,
108.4, 108, 107.3, 105, 104.7, 54.9, 43.2. MS (EI) ESþ: 574 ([Mþ þ H],
100).
4.1.5.4. N-(4-((6-((1H-Benzo[d][1,2,3]triazol-1-yl)oxy)pyrimidin-4-
yl)oxy)-3-fluorophenyl)-1-(2-methyl-4-fluorophenyl)-2-oxo-1,2-
dihydropyridine-3-carboxamide (11c). Flash chromatography using
3%e5% Ethyl acetate/DCM as eluent affords the desired product
4 .1. 6 . 2 . 1 - ( 3 , 4 - D i fl u o ro p h e nyl ) - N - ( 3 - fl u o ro - 4 - ( 6 - ( 4 -
methoxybenzylamino)pyrimidin-4-yloxy)phenyl)-2-oxo-1,2-
dihydropyridine-3-carboxamide (12b). Flash chromatography using
DCM/Ethyl acetate 70:30 as eluent affords the desired product with
with 84% yield. 1H NMR (CDCl3, 300 MHz)
d (ppm): 12.04 (s, 1H),
8.79 (dd, J1 ¼ 3 Hz, J2 ¼ 9 Hz, 1H), 8.34 (s, 1H), 8.14 (d, J ¼ 9 Hz, 1H),
7.99 (dd, J ¼ 3 Hz, J ¼ 12 Hz, 1H), 7.59e7.45 (m, 4H), 7.39 (m, 1H),
7.24e7.10 (m, 4H), 6.66 (m, 2H), 2.18 (s, 3H). 13C NMR (CDCl3,
90% yield. 1H NMR (DMSO-d6, 300 MHz)
d (ppm): 11.97 (s, 1H), 8.60
(dd, J ¼ 6 Hz, J ¼ 2 Hz, 1H), 8.14e8.11 (m, 2H), 7.93e7.85 (m, 3H),
7.70e7.66 (m, 1H), 7.46e7.17 (m, 5H), 6.90 (d, J ¼ 9 Hz, 2H), 6.75 (t,
J ¼ 6 Hz, 1H), 5.95 (s, 1H), 4.43 (s, 2H), 3.72 (s, 3H). 13C NMR (DMSO-
75 MHz) d (ppm): 170.7, 170.6, 163.5, 161.5, 161.4, 160.3, 158.1, 154.9,
151.6,145.1,144.2,142.7,137.5,137.4,135.5,129.5,129.4,128.2,125.3,
124.1, 120.1, 119.9, 117.4, 117.1, 116.2, 113.9, 113.6, 109.3, 108.3, 108,
107.2, 90.8, 17. MS (EI) ESþ: 590 ([Mþ þ Na], 100).
d6, 75 MHz) d (ppm): 168.2, 164.5, 161.6, 161.3, 160.8, 158.2, 155.3,
152, 151.3, 150.7, 150.5, 147.8, 147.4, 145, 143.8, 136.5, 136.4, 135.4,
135.2, 130.8, 128.5, 124.3, 120.2, 118, 117.7, 117.5, 117.2, 115.9, 113.7,
108.3, 108, 107, 54.9, 43.2. MS (EI) ESþ: 574 ([Mþ þ H], 100).
4.1.5.5. N-(4-((6-((1H-Benzo[d][1,2,3]triazol-1-yl)oxy)pyrimidin-4-
yl)oxy)-3-fluorophenyl)-1-(4-fluorophenyl)-2-oxo-1,2-
dihydropyridine-3-carboxamide (11d). Flash chromatography using
Cyclohexane/Ethyl acetate 65:35 as eluent affords the desired
4.1.6.3. 1-(4-Fluoro-2-methylphenyl)-N-(3-fluoro-4-(6-(4-
methoxybenzylamino) pyrimidin-4-yloxy)phenyl)-2-oxo-1,2-
dihydropyridine-3-carboxamide (12c). Flash chromatography us-
ing DCM/Ethyl acetate 90:10 as eluent affords the desired product
product with 88% yield. 1H NMR (CDCl3, 300 MHz)
d (ppm): 12.01 (s,
1H), 8.76 (dd, J ¼ 3 Hz, J ¼ 6 Hz, 1H), 8.35 (s, 1H), 8.14 (d, J ¼ 6 Hz,
1H), 7.99 (dd, J ¼ 3 Hz, J ¼ 12 Hz, 1H), 7.65e7.25 (m, 9H), 7.19 (t,
J ¼ 6 Hz, 1H), 6.67 (s, 1H), 6.65 (t, J ¼ 6 Hz, 1H). 13C NMR (CDCl3,
with 77% yield. 1H NMR (DMSO-d6, 300 MHz)
d 11.97 (s, 1H), 8.58
(dd, J ¼ 7 Hz, J ¼ 2 Hz, 1H), 8.13 (m, 2H), 7.93e7.83 (m, 3H), 7.73e
7.63 (dd, J ¼ 7 Hz, J ¼ 2 Hz, 1H), 7.48e7.38 (m, 2H), 7.25 (m, 3H),
6.90 (d, J ¼ 8 Hz, 2H), 6.73 (t, J ¼ 7 Hz, 1H), 5.95 (s, 1H), 4.43 (s,
75 MHz) d (ppm): 171.2, 164.4, 162.4, 161.4, 161.1, 158.3, 155.6, 152.3,
145.2, 143.4, 141.8, 137.7, 137.5, 135.8, 135.7, 135.1, 129, 128.5, 128.4,
128.3, 125, 123.3, 121.9, 120.6, 117, 116.7, 116.1, 109.4, 109.1, 108.6,
107.3, 90.3. MS (EI) ESþ: 576 ([Mþ þ Na], 100). MS (EI) ESꢀ: 552
([Mꢀ ꢀ H], 100). HRMS-ESI (m/z) calcd for C28H18F2N7O4 (M þ Hþ):
554.1385, found: 554.1378.
2H), 3.72 (s, 3H). 13C NMR (DMSO-d6, 75 MHz)
d (ppm): 164.5,
163.5, 161.5, 161.4, 160.2, 158.2, 157.8, 155.2, 152, 145, 144.1, 137.5,
137.4, 136.5, 135.5, 128.5, 124.3, 120.1, 117.4, 117.1, 116, 113.9, 113.7,
108.3, 108, 107.2, 54.9, 42.1, 17. MS (EI) ESþ: 570 ([Mþ þ H], 10); 592
([Mþ þ Na], 15).

798
T. Traoré et al. / European Journal of Medicinal Chemistry 70 (2013) 789e801
4 . 1. 6 . 4 . 1 - ( 4 - F l u o r o p h e n y l ) - N - ( 3 - fl u o r o - 4 - ( 6 - ( 4 -
methoxybenzylamino)pyrimidin-4-yloxy)phenyl)-2-oxo-1,2-
dihydropyridine-3-carboxamide (12d). Flash chromatography using
Cyclohexane/Ethyl acetate 60:40 as eluent affords the desired
75 MHz) d (ppm): 170.3, 168.4, 165.9, 161.6, 161.3, 158, 155.3, 152,
151.1, 150, 147.8, 147.2, 145, 143.8, 136.5, 136.4, 135.2, 124.4, 120.2,
118, 117.4, 117.2, 116, 108.3, 108, 107, 85.7, 59.7, 20.7, 14. MS (EI) ESþ:
454 ([Mþ þ H], 100). HRMS-ESI (m/z) calcd for C22H15F3N5O3
(M þ Hþ): 454.1124, found: 454.1116.
product with 94% yield. 1H NMR (CDCl3, 300 MHz)
d (ppm) 11.93 (s,
1H), 8.76 (dd, J ¼ 7 Hz, J ¼ 2 Hz, 1H), 8.25 (s, 1H), 7.93 (dd, J ¼ 12 Hz,
J ¼ 2 Hz, 1H), 7.62 (dd, J ¼ 7 Hz, J ¼ 2 Hz, 1H), 7.44e7.39 (m, 2H),
7.24e7.12 (m, 5H), 7.15 (t, J ¼ 7 Hz, 1H), 6.91e6.37 (m, 2H), 6.64 (t,
J ¼ 7 Hz, 1H), 5.84 (s, 1H), 5.31 (s, 1H), 4.43 (s, J ¼ 6 Hz, 2H), 3.82 (s,
4.1.7.3. N-(4-(6-Aminopyrimidin-4-yloxy)-3-fluorophenyl)-1-(2-
methyl-4-fluoro-phenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide
(13c). Flash chromatography using 1%e3% Ethanol/DCM as eluent
affords the desired product with 83% yield. 1H NMR (DMSO-d6,
3H). 13C NMR (DMSO-d6, 75 MHz)
d (ppm): 169, 164.4, 164.0, 163.4,
161.7, 161.1, 160.2, 158.2, 148.5, 144.6, 143.8, 136.3, 136.2, 135.3,
129.3, 128.5, 122, 120.9, 120.4, 116.1, 115.8, 113.7, 106.9, 54.9, MS (EI)
ESþ: 556 ([Mþ þ H], 60); 578 ([Mþ þ Na], 100). HRMS-ESI (m/z)
calcd for C30H24F2N5O4 (M þ Hþ): 556.1791, found: 556.1774.
300 MHz)
d
(ppm): 12.03 (s, 1H), 8.62 (d, J ¼ 6 Hz, 1H), 8.06 (d,
J ¼ 3 Hz, 1H), 8.03 (s, 1H), 7.93 (d, J ¼ 15 Hz, 1H), 7.39e7.23 (m, 5H),
6.91 (s, 2H), 6.76 (t, J ¼ 6 Hz, 1H), 5.83 (s, 1H), 2.08 (s, 3H). 13C NMR
(DMSO-d6, 75 MHz) d (ppm): 170.3, 168.4, 165.9, 163.5, 161.5, 161.4,
160.2, 158, 155.3, 152, 145, 144.1, 137.5, 136.5, 135.5, 129.5, 129.4,
124.5, 120.1, 117.4, 117.1, 116, 113.9, 113.6, 108.3, 108, 107.2, 85.7, 59.7,
20.7, 17, 14. MS (EI) ESþ: 472 ([Mþ þ H], 50). MS (EI) ESꢀ: 448
([Mꢀ þ H], 100). HRMS-ESI (m/z) calcd for C23H18F2N5O3 (M þ Hþ):
450.1374, found: 450.1397.
4.1.6.5. 1-(2-Fluoro-4-methoxyphenyl)-N-(3-fluoro-4-(6-(4-
methoxybenzylamino) pyrimidin-4-yloxy)phenyl)-2-oxo-1,2-
dihydropyridine-3-carboxamide (12e). Flash chromatography us-
ing 1%e2% Ethanol/DCM as eluent affords the desired product with
83% yield. 1H NMR (DMSO-d6, 300 MHz)
d (ppm): 11.94 (s, 1H), 8.60
(dd, J ¼ 7 Hz, J ¼ 2 Hz, 1H), 8.13 (dd, J ¼ 7 Hz, J ¼ 2 Hz, 2H), 7.91 (d,
J ¼ 12 Hz, 2H), 7.54 (t, J ¼ 8 Hz, 1H), 7.40 (d, J ¼ 8.0 Hz, 1H), 7.24 (d,
J ¼ 8 Hz, 3H), 7.13 (dd, J ¼ 12 Hz, J ¼ 2 Hz, 1H), 6.97 (dd, J ¼ 8 Hz,
J ¼ 2 Hz, 1H), 6.89 (d, J ¼ 8 Hz, 2H), 6.74 (t, J ¼ 7 Hz, 1H), 5.94 (s, 1H),
4.43 (s, 2H), 3.85 (s, 3H), 3.72 (s, 3H). 13C NMR (DMSO-d6, 75 MHz)
4.1.7.4. N-(4-(6-Aminopyrimidin-4-yloxy)-3-fluorophenyl)-1-(4-
fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide
(13d).
Flash chromatography using 2%e3% Ethanol/DCM as eluent affords
the desired product with 68% yield. 1H NMR (DMSO-d6, 300 MHz)
d
(ppm): 12.03 (s, 1H), 8.57 (d, J ¼ 7 Hz, 1H), 8.19e7.98 (m, 2H), 7.90
d
(ppm): 164.5, 161.4, 161.2, 161.1, 158.7, 158.2, 155.4, 155.2, 152,
(d, J ¼ 12 Hz, 1H), 7.71e7.51 (m, 2H), 7.41 (t, J ¼ 8 Hz, 3H), 7.26 (t,
145.2, 144.6, 136.5, 136.3, 135.2, 129.5, 128.5, 124.3, 120, 119.8, 116,
113.7, 110.7, 108.3, 108, 107.1, 102.4, 102.1, 56, 54.9, 43.2 MS (EI) ESþ:
586 ([Mþ þ H], 50).
J ¼ 8 Hz, 1H), 6.90 (s, 2H), 6.71 (t, J ¼ 7 Hz, 1H), 5.83 (s, 1H). 13C NMR
(DMSO-d6, 75 MHz)
d (ppm): 168.4, 165.9, 163.4, 161.7, 161.4, 160.2,
158, 155.3, 152, 144.9, 144.1, 136.4, 136.2, 136.1, 129.3, 129.2, 124.4,
120.1, 116.2, 116, 115.8, 108.3, 108, 106.9, 85.7. MS (EI) ESþ: 458
([Mþ þ H], 100). MS (EI) ESꢀ: 434 ([Mꢀ þ H], 100). HRMS-ESI (m/z)
calcd for C22H16F2N5O3 (M þ Hþ): 436.1218, found: 436.1216.
4.1.7. General procedure E for the synthesis of N-(4-(6-
aminopyrimidin-4-yloxy)-3-fluorophenyl)-1-(substituted-phenyl)-
2-oxo-1,2-dihydropyridine-3-carboxamide (13aee)
In a dry flask under argon, to a solution of the corresponding
compound 12 (0.22 mmol) in DCM (1 mL) was added dropwise TFA
(3 mL) and the mixture was heated at reflux during 6 h. The solvent
was removed, the residue was extracted with EtOAc, washed with
basic water until neutral pH, then with brine, dried over MgSO4 and
filtered. The solvent was evaporated to give the crude product
which was purified by flash chromatography to afford the desired
product.
4.1.7.5. N-(4-(6-Aminopyrimidin-4-yloxy)-3-fluorophenyl)-1-(2-
fluoro-4-methoxyphenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide
(13e). Flash chromatography using 1%e3% Ethanol/DCM as eluent
affords the desired product with 91% yield. 1H NMR (DMSO-d6,
300 MHz)
d
(ppm): 11.94 (s,1H), 8.59 (dd, J ¼ 7 Hz, J ¼ 2 Hz,1H), 8.12
(dd, J ¼ 7 Hz, J ¼ 2 Hz, 1H), 8.04 (d, J ¼ 1 Hz, 1H), 7.91 (dd, J ¼ 12 Hz,
J ¼ 2 Hz, 1H), 7.53 (t, J ¼ 9 Hz, 1H), 7.45e7.35 (m, 1H), 7.27 (t,
J ¼ 9 Hz, 1H), 7.12 (dd, J ¼ 12 Hz, J ¼ 2 Hz, 1H), 7.00e6.84 (m, 3H),
6.78e6.67 (m, 1H), 5.85 (d, J ¼ 1 Hz, 1H), 3.84 (s, 3H). 13C NMR
4.1.7.1. N-(4-(6-Aminopyrimidin-4-yloxy)-3-fluorophenyl)-1-(2,4-
(DMSO-d6, 75 MHz) d (ppm): 168.4, 166, 161.4, 161.3, 161.2, 161.1,
difluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide
(13a).
158.7, 158, 155.4, 155.3, 152, 145.2, 144.6, 136.5, 136.4, 135.4, 135.2,
129.5, 124.4, 120, 119.8, 116.1, 110.7, 108.4, 108.1, 107.1, 102.4, 102.1,
85.7, 56, 55.9, 54.8, 18.5. MS (EI) ESþ: 466 ([Mþ þ H], 100). MS (EI)
ESþ: 466 ([Mþ þ H], 100). HRMS-ESI (m/z) calcd for C23H18F2N5O4
(M þ Hþ): 466.1323, found: 466.1313.
Flash chromatography using 2%e5% Ethanol/DCM as eluent affords
the desired product with 74% yield. 1H NMR (DMSO-d6, 300 MHz)
d
(ppm): 11.85 (s, 1H), 8.62 (dd, J ¼ 7 Hz, J ¼ 2 Hz, 1H), 8.16 (dd,
J ¼ 7 Hz, J ¼ 2 Hz, 1H), 8.05 (s, 1H), 7.92 (dd, J ¼ 12 Hz, J ¼ 2 Hz, 1H),
7.84e7.68 (m, 1H), 7.61 (td, J ¼ 12 Hz, J ¼ 2 Hz, 1H), 7.34 (m, 3H),
6.93 (s, 2H), 6.77 (t, J ¼ 7 Hz, 1H), 5.86 (s, 1H). 13C NMR (DMSO-d6,
4.1.8. N-(3-Fluoro-4-(6-(2-Morpholinoethylamino)pyrimidin-4-
yloxy)phenyl)-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-
carboxamide (14)
75 MHz)
d (ppm): 168.4, 166, 164.1, 164, 161.2, 161.1, 160.8, 160.7,
158.6, 158.4, 158, 155.3, 155, 152, 145.5, 144.2, 136.4, 136.3, 135.4,
135.3, 130.7, 124.4, 124, 123.9, 123.8, 120.1, 116.1, 112.5, 112.2, 108.4,
108.1, 107.3, 105.4, 105.1, 104.7. MS (EI) ESþ: 454 ([Mþ þ H], 100).
HRMS-ESI (m/z) calcd for C22H15F3N5O3 (M þ Hþ): 454.1124, found:
454.1144.
In a dry flask under argon, to a solution of the benzotriazol 11d
(0.1 g, 0.18 mmol), and cesium carbonate (0.11 g, 0.36 mmol) in
DME (3 mL) was added 2-aminoethyl-morpholine (0.03 mL,
0.21 mmol) and the mixture was heated at reflux during 6 h. The
solvent was removed, the residue was extracted with EtOAc,
washed with water, brine, dried over MgSO4 and filtered. The sol-
vent was evaporated to give the crude product which was purified
by flash chromatography using 1% Ethanol/DCM as eluent to afford
the desired product with 76% yield. 1H NMR (DMSO-d6, 300 MHz)
4.1.7.2. N-(4-(6-Aminopyrimidin-4-yloxy)-3-fluorophenyl)-1-(3,4-
difluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide
(13b).
Flash chromatography using 2%e5% Ethanol/DCM as eluent affords
the desired product with 55% yield. 1H NMR (DMSO-d6, 300 MHz)
d
(ppm): 11.97 (s, 1H), 8.58 (dd, J ¼ 7 Hz, J ¼ 2 Hz, 1H), 8.13 (dd,
d
(ppm): 12.04 (s, 1H), 8.57 (dd, J ¼ 7 Hz, J ¼ 2 Hz, 1H), 8.11 (dd,
J ¼ 7 Hz, J ¼ 2 Hz, 1H), 8.04 (s, 1H), 7.98e7.78 (m, 2H), 7.68 (dd,
J ¼ 20 Hz, J ¼ 9 Hz, 1H), 7.54e7.34 (m, 2H), 7.27 (t, J ¼ 9 Hz, 1H), 6.92
(s, 2H), 6.73 (t, J ¼ 7 Hz, 1H), 5.85 (s, 1H). 13C NMR (DMSO-d6,
J ¼ 7 Hz, J ¼ 2 Hz, 2H), 7.91 (d, J ¼ 12 Hz, 1H), 7.60 (dd, J ¼ 9 Hz,
J ¼ 4 Hz, 2H), 7.33 (dt, J ¼ 47 Hz, J ¼ 9 Hz, Hz, 5H), 6.71 (t, J ¼ 7 Hz,
1H), 5.95 (s, 1H), 3.55 (d, J ¼ 4 Hz, 4H), 2.60e2.23 (m, 8H). 13C NMR

T. Traoré et al. / European Journal of Medicinal Chemistry 70 (2013) 789e801
799
(DMSO-d6, 75 MHz)
d
(ppm): 164.5, 163.4, 161.7, 161.4, 160.2, 155.3,
as a brown solid. This residue was purified by flash chromatography
using Cyclohexane/Ethyl Acetate 65:35 as eluent to afford the
desired product as a white solid with 96% yield. 1H NMR (DMSO-d6,
152, 144.9, 144.1, 136.2, 135.4, 135.2, 129.3, 129.2, 124.4, 120, 116.2,
116, 115.8, 112.1, 108.3, 108, 106.9, 66.1, 53.2. MS (EI) ESþ: 549
([Mþ þ H], 50); 571 ([Mþ þ Na], 100). HRMS-ESI (m/z) calcd for
300 MHz)
(s, 1H), 6.55e6.25 (m, 2H), 6.11 (d, J ¼ 6 Hz, 1H), 5.34 (s, 2H), 4.33 (s,
1H), 3.69 (s, 3H). 13C NMR (DMSO-d6, 75 MHz)
(ppm) 169.3, 162.1,
d
(ppm): 8.09 (d, J ¼ 6 Hz, 1H), 6.93 (t, J ¼ 9 Hz, 1H), 6.79
C
28H27F2N6O3 (M þ Hþ): 549.2056, found: 549.2053.
d
4.1.9. N-(3-Fluoro-4-(6-(methylaminopyrimidin-4-yloxy)phenyl)-
1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide) (15)
In a dry flask under argon, to a solution of the benzotriazol 11d
(0.08 g, 0.17 mmol), and cesium carbonate (0.114 g, 0.35 mmol) in
DME (1.5 mL) was added methylamine NH2Me$HCl (0.018 g,
0.26 mmol) and the mixture was heated at reflux during 6 h. The
solvent was removed, the residue was extracted with EtOAc,
washed with water, brine, dried over MgSO4 and filtered. The sol-
vent was evaporated to give the crude product which was purified
by flash chromatography using Cyclohexane/Ethyl Acetate 40:60 as
eluent to afford the desired product with 55% yield. 1H NMR
157.9, 148, 132, 128.4, 124.1, 113.3, 109.4, 54.9, 43.3, 40.2, 40.1, 39.1,
38.9, 38.6. MS (EI) ESþ: 341 ([Mþ þ H], 100); 363 ([Mþ þ Na], 40).
4.1.13. 4-(2-Fluoro-4-nitrophenoxy)-N-methylpyrimidin-2-amine
(18b)
In a dry sealed tube under argon were placed the compound
17 (0.67 g, 2.4 mmol), MeNH2$HCl (0.2 g, 2.9 mmol), potassium
carbonate (0.68 g, 4.9 mmol) in DMF (11 mL) and the mixture
was heated at 60 ꢁC during 15 h. The mixture was then cooled at
room temperature and the solvent evaporated under high vac-
uum. The residue was extracted with EtOAc, washed with a
saturated solution of NaHCO3, water and brine. The organic layers
were dried over MgSO4, filtered and the solvent was evaporated
to give the crude product as a yellow solid. This residue was
purified by flash chromatography using Cyclohexane/Ethyl ace-
tate 90:10 as eluent to afford the desired product as a light yel-
(DMSO-d6, 300 MHz)
d
(ppm): 12.04 (s, 1H), 8.58 (d, J ¼ 6 Hz, 1H),
8.21e7.98 (m, 2H), 7.91 (d, J ¼ 12 Hz, 1H), 7.72e7.51 (m, 2H), 7.51e
7.17 (m, 5H), 6.72 (t, J ¼ 6 Hz, 1H), 5.91 (s, 1H), 2.78 (s, 3H). 13C NMR
(DMSO-d6, 75 MHz)
d (ppm): 165.2, 163.4, 161.7, 161.4, 160.1, 160.2,
155.3, 152, 144.9, 144.1, 136.4, 136.2, 135.4, 135.2, 129.3, 129.2, 124.3,
120.1,116.2,115.9,108.3,108,106.9, 95.2, 93.4, 28.9. MS (EI) ESþ: 472
([Mþ þ Na], 100). HRMS-ESI (m/z) calcd for C23H18F2N5O3 (M þ Hþ):
450.1374, found: 450.1366.
low solid with 16% yield. 1H NMR (DMSO-d6, 300 MHz)
8.45e8.07 (m, 3H), 7.69 (t, J ¼ 9 Hz, 1H), 6.36 (s, 1H), 2.50 (s, 3H).
d (ppm):
13C NMR (DMSO-d6, 75 MHz)
d (ppm): 167.6, 162.5, 160.5, 155,
151.6, 145.2, 145, 125, 120.7, 112.9, 112.6, 96, 27.7. MS (EI) ESþ: 265
4.1.10. 2-Chloro-4-(2-fluoro-4-nitrophenoxy)pyrimidine (17)
This compound was obtained with 58% yield according to the
method described for compound 5a. 1H NMR (DMSO-d6, 300 MHz)
([Mþ þ H], 100).
4.1.14. 4-(4-Amino-2-fluorophenoxy)-N-methylpyrimidin-2-amine
(19b)
d
(ppm): 8.76 (d, J ¼ 6 Hz, 1H), 8.43 (dd, J ¼ 9 Hz, J ¼ 3 Hz, 1H), 8.23
(ddd, J1 ¼9 Hz, J ¼ 3 Hz, J ¼ 2 Hz,1H), 7.80 (dd, J ¼ 9 Hz, J ¼ 8 Hz,1H),
This compound was obtained with a quantitative yield accord-
7.48 (d, J ¼ 6 Hz, 1H). 13C NMR (CDCl3, 75 MHz)
d (ppm): 168.4,
ing to the method described for compound 19a. 1H NMR (DMSO-d6,
160.9, 160.4, 155.3, 151.9, 144.4, 144.2, 124.2, 120.5, 120.4, 113.4,
300 MHz)
d
(ppm): 8.11 (s, 1H), 7.0 (s, 1H), 6.93 (t, J ¼ 9 Hz, 1H), 6.39
113.1, 106.8 MS (EI) ESþ: 270 ([Mþ þ H], 90); 282 ([Mþ þ Na], 100).
(dd, J ¼ 24 Hz, J ¼ 9 Hz, 2H), 6.08 (s, 1H), 5.31 (s, 2H), 2.66 (s, 3H). 13
C
NMR (DMSO-d6, 75 MHz)
d (ppm): 169.9, 163.1, 159.4, 156.6, 153.3,
4.1.11. 4-(2-Fluoro-4-nitrophenoxy)-N-(4-methoxybenzyl)
pyrimidin-2-amine (18a)
145.4, 145.3, 131.4, 131.3, 124.3, 124.2, 110.5, 110.4, 103.4, 103.1, 28.2.
MS (EI) ESþ: 265 ([Mþ þ H], 100).
In a dry sealed tube under argon were placed the compound 17
(0.9 g, 3.3 mmol), 4-methoxy-benzylamine (0.6 mL, 4 mmol), po-
tassium carbonate (0.54 g, 4 mmol) in DMF (15 mL) and the mixture
was heated at 60 ꢁC for 30 min. The mixture was then cooled at
room temperature and the solvent evaporated under high vacuum.
The residue was extracted with EtOAc, washed with a saturated
solution of NaHCO3, water and brine. The organic layers were dried
over MgSO4, filtered and the solvent was evaporated to give the
crude product as a yellow solid. This residue was purified by flash
chromatography using Cyclohexane/Ethyl acetate 90:10 as eluent
to afford the desired product as a light yellow solid with 26% yield.
4.1.15. N-(3-Fluoro-4-(2-(4-Methoxybenzylamino)pyrimidin-4-
yloxy)phenyl)-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-
carboxamide (20a)
This compound was obtained with 85% yield according to the
general procedure C. 1H NMR (CD2Cl2, 300 MHz)
d (ppm): 12.04 (s,
1H), 8.72 (d, J ¼ 7 Hz, 1H), 8.17 (s, 1H), 7.95 (d, J ¼ 12 Hz, 1H), 7.67 (d,
J ¼ 5 Hz, 1H), 7.45e7.41 (m, 2H), 7.31e7.26 (m, 3H), 7.17 (t, J ¼ 8 Hz,
2H), 6.83 (d, J ¼ 8 Hz, 2H), 6.65 (t, J ¼ 8 Hz,1H), 6.24 (d, J ¼ 5 Hz,1H),
5.39 (m, 2H), 3.78 (s, 3H), 2.80 (s, 2H). 13C NMR (CDCl3, 75 MHz)
d
(ppm): 169.3, 164.4, 162.4, 162.1, 161.3, 161, 159.5, 158.7, 156, 152.7,
1H NMR (DMSO-d6, 300 MHz)
1H), 7.16 (s, 1H), 6.79 (s, 3H), 6.37 (s, 1H), 4.34 (s, 1H), 3.96 (s, 1H),
3.69 (s, 3H). 13C NMR (CDCl3, 75 MHz)
(ppm): 168.2, 161.9, 161.2,
d
(ppm): 8.43e8.08 (m, 2H), 7.66 (s,
145.1, 141.6, 136.7, 136.6, 135.8, 135.7, 130.9, 129, 128.4, 128.3, 123.9,
122.1, 117, 116.7, 115.8, 113.8, 109.2, 108.9, 107.2, 55.2, 44.8. MS (EI)
ESþ: 556 ([Mþ þ H], 100). HRMS-ESI (m/z) calcd for C30H24F2N5O4
(M þ Hþ): 556.1791, found: 556.1781.
d
158.8, 155.6, 152.3, 145.7, 145.5, 145.2, 130.5, 128.6, 124.5, 124.4, 120,
119.9, 113.8, 112.9, 112.6, 99.9, 55.2, 44.8. MS (EI) ESþ: 370
([Mþ þ H], 100).
4.1.16. N-(4-(2-Aminopyrimidin-4-yloxy)-3-fluorophenyl)-1-(4-
fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide (21)
This compound was obtained with 85% yield according to the
4.1.12. 4-(4-Amino-2-fluorophenoxy)-N-(4-methoxybenzyl)
pyrimidin-2-amine (19a)
general procedure E. 1H NMR (CD2Cl2, 300 MHz)
d (ppm): 12.04 (s,
In a dry flask under argon, the compound 18a (0.2 g, 0.54 mmol)
was dissolved in a mixture of THF/MeOH 1:1 (15 mL), then Zn
(0.35 g, 5.4 mmol) and NH4Cl (0.28, 5.4 mmol) were added. The
mixture was stirred at room temperature during 1 h. The crude was
then filtered over a pad of celite, rinsed with EtOAc and the filtrate
was evaporated. The brown residue was extracted with EtOAc. The
organic layer was washed with water and brine, dried over MgSO4
and filtered. The solvent was evaporated to give the crude product
1H), 8.74 (dd, J ¼ 7 Hz, J ¼ 2 Hz, 1H), 8.16 (d, J ¼ 6 Hz, 1H), 7.92 (dd,
J ¼ 12 Hz, J ¼ 2 Hz, 1H), 7.67 (dd, J ¼ 7 Hz, J ¼ 2 Hz, 1H), 7.46e7.37
(m, 2H), 7.33e7.21 (m, 4H), 7.18 (t, J ¼ 8 Hz,1H), 6.65 (t, J ¼ 7 Hz,1H),
6.29 (d, J ¼ 6 Hz, 1H), 4.96 (s, 2H). 13C NMR (DMSO-d6, 75 MHz)
d
(ppm): 168.8, 163.5, 161.7, 161.5, 160.2, 160.1, 155.2, 151.9, 144.9,
144.1, 136.6, 136.2, 135.1, 129.3, 129.2, 124.4, 120.1, 116.2, 116.1, 115.9,
108.4, 108.1, 106.9, 95.3 MS (EI) ESþ: 436 ([Mþ þ H], 100). HRMS-ESI
(m/z) calcd for C22H16F2N5O3 (M þ Hþ): 436.1218, found: 436.1228.

800
T. Traoré et al. / European Journal of Medicinal Chemistry 70 (2013) 789e801
4.1.17. N-(3-Fluoro-4-(2-(methylamino)pyrimidin-4-yloxy)
phenyl)-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-
carboxamide (20b)
plate was incubated at room temperature until the blue color
emerged. The reaction was stopped by adding 100 l of 2 M H2SO4
and the absorbance was read at 450 nm using a multi-well
spectrophotometer.
Kinase assays on a small panel of kinases for a subset of syn-
thesized molecules were carried out at KinomScan, a division of
DiscoveRex, 11180 Roselle St. Suite D, San Diego, CA 92121.
Compound 1 (BMS-777607) was purchased from Selleckchem
(München, Germany).
m
This compound was obtained with 40% yield according to the
general procedure C. 1H NMR (CD2Cl2, 300 MHz)
d
(ppm): 12.03 (s,
1H), 8.72 (dd, J ¼ 7 Hz, J ¼ 2 Hz, 1H), 8.16 (d, J ¼ 5 Hz, 1H), 7.92 (dd,
J ¼ 12 Hz, J ¼ 2 Hz, 1H), 7.67 (dd, J ¼ 7 Hz, J ¼ 2 Hz, 1H), 7.44 (dd,
J ¼ 9 Hz, J ¼ 5 Hz, 2H), 7.30 (dd, J ¼ 14 Hz, J ¼ 6 Hz, 3H), 7.16 (t,
J ¼ 9 Hz, 1H), 6.63 (t, J ¼ 7 Hz, 1H), 6.18 (d, J ¼ 5 Hz, 1H), 2.85 (s, 3H).
13C NMR (DMSO-d6, 75 MHz)
d (ppm): 168.5, 163.4, 162.5, 161.7,
161.5, 161.3, 160.2, 159.9, 155.2, 152, 144.8, 144.1, 136.7, 136.5, 136.2,
136.1, 129.3, 129.2, 124.4, 120.1, 116.2, 115.9, 108.1, 106.9, 99.4, 27.7.
MS (EI) ESþ: 450 ([Mþ þ H], 100). HRMS-ESI (m/z) calcd for
4.3. Molecular modeling
Experimental crystallographic coordinates of human Met (PDB
ID: 3F82, resolution 2.50 A) and a DFG-out model of Tyro3 were
C
23H18F2N5O3 (M þ Hþ): 450.1374, found: 450.1371.
ꢀ
used in the docking studies.
4.1.18. N-(4-(6-(1H-Benzo[d][1,2,3]triazol-1-yloxy)pyrimidin-4-
yloxy)phenyl)-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-
carboxamide (22)
Docking experiments were performed with FRED version 2.2.5
[26], OpenEye Scientific Software, Santa Fe, NM. http://eyesopen.
com, using Chemgauss3 scoring function [27]. A conformational
database of listed compounds was generated with OMEGA version
2.3.2 [28] and subsequently used as input to FRED. The required
receptor file was built with FRED_Receptor using multiple molec-
ular probes detection. No constraint was used in order not to
restrict the pose selection. The best ranked pose for each compound
was retained and analyzed in order to clarify the interactions
involved in the ligand binding.
This compound was obtained with 72% yield according to the
general procedure C. 1H NMR (CDCl3, 300 MHz)
d (ppm): 11.92 (s,
1H), 8.80e8.71 (m, 1H), 8.34 (s, 1H), 8.08 (d, J ¼ 9 Hz, 1H), 7.80 (d,
J ¼ 9 Hz, 2H), 7.61 (dd, J ¼ 7 Hz, J ¼ 2 Hz, 1H), 7.58 (d, J ¼ 7 Hz, 1H),
7.47 (dd, J ¼ 8 Hz, J ¼ 5 Hz, 2H), 7.49e7.45 (m, 2H), 7.25 (dd,
J ¼ 12 Hz, J ¼ 5 Hz, 2H), 7.14 (d, J ¼ 9 Hz, 2H), 6.58 (dd, J ¼ 12 Hz,
J ¼ 5 Hz, 2H). 13C NMR (CDCl3, 75 MHz)
d (ppm): 172.1, 171.2, 164.3,
162.4, 161.3, 161, 158.4, 148, 145.1, 143.3, 141.6, 136.5, 135.9, 135.8,
128.9, 128.4, 128.3, 122.1, 121.8, 121.7, 116.9, 116.6, 108.5, 107.2, 90.3.
MS (EI) ESþ: 558 ([Mþ þ H], 30). MS (EI) ESꢀ: 534 ([Mꢀ ꢀ H], 30).
Acknowledgments
The authors would like to thank « L’Institut National du Cancer »
for post-doctoral fellowships for TT and AC. This study was sup-
ported by « L’Institut National du Cancer ». (Translational Research
2011 Program). The authors thank Mariane Bombled for performing
mass spectra.
4.1.19. N-(4-(6-Aminopyrimidin-4-yloxy)phenyl)-1-(4-
fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide (23)
This compound was obtained according to the general proce-
dure D (62% yield) and E (96% yield). 1H NMR (DMSO-d6, 300 MHz)
d
(ppm): 11.94 (s, 1H), 8.58 (dd, J ¼ 7 Hz, J ¼ 2 Hz, 1H), 8.19e7.98 (m,
2H), 7.74 (d, J ¼ 9 Hz, 2H), 7.66e7.55 (m, 2H), 7.48e7.35 (m, 2H), 7.13
(d, J ¼ 9 Hz, 2H), 6.84 (s, 2H), 6.71 (t, J ¼ 7 Hz, 1H), 5.71 (s, 1H). 13
C
Appendix A. Supplementary data
NMR (DMSO-d6, 75 MHz)
d (ppm): 169.4, 165.9, 163.4, 161.7, 161.1,
Supplementary data related to this article can be found at http://
dx.doi.org/10.1016/j.ejmech.2013.10.037.
160.2, 158.1, 148.5, 144.7, 143.8, 136.2, 135.3, 129.3, 129.2, 122, 120.9,
120.4, 116.1, 115.8, 106.9, 86.2. MS (EI) ESþ: 418 ([Mþ þ H], 100).
HRMS-ESI (m/z) calcd for C22H17FN5O3 (M þ Hþ): 418.1312, found:
418.1335.
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