Synthesis of aryl phosphates based on pyrimidine and triazine scaffolds

Article abstract of DOI:10.1016/j.ejmech.2009.10.003

The syntheses of the triazinyl-based bis-aryl phosphates 2 and 3, and of the aminopyrimidyl-based aryl phosphate 4 are described. Each compound contains a diaryl ether-phosphate structural motif. The synthetic route to bis-aryl phosphates 2 and 3 consisted in two nucleophilic substitution reactions with amines from cyanuric chloride, followed by a Suzuki coupling with the resulting 2,4-diamino-6-chloro-1,3,5-triazine derivative 12 to introduce the diaryl ether functionality. Aryl phosphate 4 was obtained via condensation of aryl guanidine 34 with aryloxyphenyl butenone 31. These de novo-designed aryl phosphates were evaluated as potential inhibitors of the Grb2-SH2 domain using an ELISA assay. The water-soluble sodium salt 26 of 3 gave an IC₅₀ value in the high micromolar range. Molecular modeling studies were subsequently performed upon modifying the 1,3,5-trisubstituted triazine scaffold of 3. Non-phosphate derivatives encompassing cyclopropane, pyrrole, keto-acid, and IZD fragments were thus step-wise designed and their Grb2-SH2 complexes were modeled by molecular dynamics. Some derivatives gave rise to an enriched pattern of H-bonds and cation-π interactions with Grb2-SH2.

Full text of DOI:10.1016/j.ejmech.2009.10.003

European Journal of Medicinal Chemistry 45 (2010) 244–255
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Synthesis of aryl phosphates based on pyrimidine and triazine scaffolds
,
,
,
,
,d
Caroline Courme a b, Nohad Gresh c d, Michel Vidal c d, Christine Lenoir c d, Christiane Garbay c
,
Jean-Claude Florent a b, Emmanuel Bertounesque a
,
,b,
*
a CNRS UMR 176, 26 rue d’Ulm, 75005 Paris, France
b Institut Curie, Centre de Recherche, 26 rue d’Ulm, 75005 Paris, France
c Universite´ Paris Descartes, UFR Biome´dicale, 45 rue des Saints-Pe`res, 75006 Paris, France
d INSERM U648, Laboratoire de Pharmacochimie Mole´culaire et Cellulaire, Paris, France
a r t i c l e i n f o
a b s t r a c t
Article history:
The syntheses of the triazinyl-based bis-aryl phosphates 2 and 3, and of the aminopyrimidyl-based aryl
phosphate 4 are described. Each compound contains a diaryl ether-phosphate structural motif. The
synthetic route to bis-aryl phosphates 2 and 3 consisted in two nucleophilic substitution reactions with
amines from cyanuric chloride, followed by a Suzuki coupling with the resulting 2,4-diamino-6-chloro-
1,3,5-triazine derivative 12 to introduce the diaryl ether functionality. Aryl phosphate 4 was obtained via
condensation of aryl guanidine 34 with aryloxyphenyl butenone 31. These de novo-designed aryl
phosphates were evaluated as potential inhibitors of the Grb2-SH2 domain using an ELISA assay. The
water-soluble sodium salt 26 of 3 gave an IC50 value in the high micromolar range. Molecular modeling
studies were subsequently performed upon modifying the 1,3,5-trisubstituted triazine scaffold of 3. Non-
phosphate derivatives encompassing cyclopropane, pyrrole, keto-acid, and IZD fragments were thus
step-wise designed and their Grb2-SH2 complexes were modeled by molecular dynamics. Some deriv-
Received 9 December 2008
Received in revised form
25 September 2009
Accepted 1 October 2009
Available online 9 October 2009
Keywords:
2-Aminopyrimidines
Aryl phosphates
Heck reaction
Suzuki–Miyaura reaction
1,3,5-Triazines
atives gave rise to an enriched pattern of H-bonds and cation–
p interactions with Grb2-SH2.
Protein–tyrosine kinases
Grb2-SH2 domain
De novo design
Ó 2009 Elsevier Masson SAS. All rights reserved.
1. Introduction
pharmacophores in the design of cell signalling inhibitors as
potential anticancer agents [17–22].
The 2-arylaminopyrimidine and 1,3,5-triazine motifs are
important as scaffolds in drug discovery chemistry. 2-Arylamino-
pyrimidine derivatives have exhibited biological activities such as
antitumor activities involving different targets (e.g. CDKs, VEGF–
TKRs, Bcr–Abl kinase) [1–6], and antiinflammatory activity by
inhibition of Lck [7]. 1,3,5-Triazine derivatives [8] have displayed
a broad range of biological activities including cytotoxic activities
[9–11], antiangiogenic activity by targeting either VEGF-R2 (KDR)
[12] or direct modulation of Tie-2 tyrosine kinase phosphorylation
[13], antiparasitic activities [14,15], and glucocerebrosidase inhibi-
tion with potential as chemical chaperones for Gaucher disease
[16]. To our knowledge, the syntheses of aryl phosphates based on
pyrimidine and triazine scaffolds have not been reported to date.
Importantly, the aryl phosphate group or its mimics are key
As part of our efforts in the synthesis of non-peptidic inhibitors
of the SH2 domain (Src homology) of Grb2 [23] (growth factor
receptor-bound protein-2) which mediates protein–protein inter-
actions in tyrosine kinase signal transduction pathways [17–22], we
have focused on the design and synthesis of aryl phosphates based
on heterocycles allowing diverse functionalization [24]. Structure-
based de novo design was carried out using 1 as a reference ligand
[25–27], encompassing three pharmacophores [i.e., pTyr, (aMe)p-
Tyr, CONH2] for binding to Grb2 (IC50 ¼ 11 nM, ELISA assay). The
complexes of two pseudopeptides with the Grb2-SH2 domain were
recently reported by Martin and co-workers [28–30].
The structure of the best MD [25] pose of 1 with Grb2-SH2 is
presented in the Supporting Information. Using this structure, 2
and 3 were designed by computer graphics upon replacing the
peptide scaffold by
a 1,3,5-trisubstituted triazine one while
retaining a satisfactory overlap of the phosphate groups at the two
extremities with those of 1. Within this context, we describe here
the syntheses of the triazinyl-containing bis-aryl phosphates 2 and
3, and the synthesis of the 2-arylaminopyrimidyl-containing aryl
phosphate 4 to mimic [31,32] the phosphate moiety of 2. These
* Corresponding author. CNRS UMR 176, 26 rue d’Ulm, 75005 Paris, France. Tel.:
þ33 156246659; fax: þ33 156246631.
E-mail address: emmanuel.bertounesque@curie.fr (E. Bertounesque).
0223-5234/$ – see front matter Ó 2009 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejmech.2009.10.003

C. Courme et al. / European Journal of Medicinal Chemistry 45 (2010) 244–255
245
N
N
NH
N
OPO3H2
OPO3H2
O
NH2
Asn
O
H
N
H
N
H2N
HN
HN
N
CONH2
HN
N
HN
N
CONH2
OPO3H2
HN
O
N
N
N
N
N
O
pTyr
O
H2O3PO
H2O3PO
N
H
mAZ
(αMe)pTyr
Me
H2O3PO
H2O3PO
O
O
1
2
3
4
Fig. 1.
novel compounds have in common a diaryl ether-phosphate
structural motif (Fig. 1).
satisfying in terms of yields. As expected, the phenol group of 4-
aminophenol is unreactive under these reaction conditions [69].
In order to install the diaryl ether fragment on 12 via Suzuki
coupling, the meta- and ortho-substituted boronic acids 15 and
18 were prepared from the corresponding brominated deriva-
tives. The direct SNAr displacement [70] of 1-bromo-3-fluo-
robenzene with hydroquinone, using sodium methoxide as base,
gave aryloxyphenol 13 [71] in 43% yield [72] (Scheme 2). After
benzylation, halogen–lithium exchange of 1-(4-benzylox-
yphenoxy)-3-bromobenzene 14, subsequent reaction of the anion
with triisopropylboron and acidic hydrolysis led to 15 [73] in 81%
yield. The ortho-substituted boronic acid 18 was prepared
following the same procedure.
Importantly, the common substructure of 2 and 3 [33,34],
namely 4-(4-amino-1,3,5-triazin-2-ylamino)phenyl phosphate,
was only published in recent patents (5–7) [35] reporting inhibition
of smooth cell proliferation, treatment of inflammation, hyper-
proliferation, and modulation of glycosidase (Fig. 2). As for the
substructure of 4, namely N-(4-(2H-tetrazol-5-yl)phenyl)pyr-
imidin-2-amine, it also appeared in the three patented compounds
8–10 [36–38], in the context of the syntheses of protein kinase
inhibitors useful in treatment of diseases like cancer.
2. Results and discussion
The Suzuki coupling of 12 with 15 is depicted in Scheme 3.
Palladium tetrakis(triphenylphosphine) was chosen as the catalyst
as it is the most commonly used for Suzuki couplings with chloro-
triazines [56–59]. It appeared that the best solvent was a mixture of
acetonitrile and water thoroughly degassed under high vacuum.
Those conditions afforded 19 in 67% yield. It is worth pointing out
that the same solvent degassed only via argon bubbling gave
mostly homocoupling product and a very small amount of the
desired Suzuki product. Toluene/water [56] and DME/water
mixtures did not lead either to the expected results. The same
reaction conditions were used for the ortho-substituted boronic
acid 18, giving compound 21 in 51% yield. Both Suzuki adducts 19
and 21 were debenzylated by hydrogenation with H2 over Pd/C to
give, respectively, 20 and 22 in good yields.
As outlined in Scheme 4, the resulting diphenols 20 and 22 were
then converted [74,75], respectively, into the corresponding bis-
dibenzyl phosphates 23 (79%) and 24 (69%). The latter were
hydrogenated over Pd/C to give the non-water soluble bis-aryl
phosphates 2 and 3, of very low solubility in usual solvents except
in DMSO. However, their limited solubilities necessited a quantity
of DMSO of 10% final volume in PBS buffer. For the range of
concentrations to be studied, this does not enable reliable ELISA-
The syntheses of the triazine scaffold-based bis-aryl phosphates
2 and 3 were carried out starting from 2,4,6-trichloro-1,3,5-triazine
(cyanuric chloride) [8]. The latter and its derivatives (i.e., mono- or
dichloride) are known to react very easily with various nucleo-
philes such as amines [9–11,15,39–46], alcohols [47,48], thiols
[49,50], and cyanide [51,52]. It can also undergo organometallics
couplings such as the Stille [53,54], Suzuki–Miyaura [55–59] or
Negishi [54,60] reactions, as well as the Grignard alkylation [61].
The trifunctionalization of cyanuric chloride to prepare the tri-
azinyl-based bis-aryl phosphates 2 and 3 is shown in Fig. 3. First, 2-
aminoacetamide and 4-aminophenol would be introduced by
a stepwise nucleophilic substitution of two chloro groups. Then,
installation of the meta- or ortho-diaryl ether unit would be carried
out via Suzuki coupling [62–68].
Thus, glycinamide hydrochloride was coupled to cyanuric
chloride at 0 ꢀC, the resulting product 11 was then reacted with
4-aminophenol at room temperature to give the 2,4-diamino-6-
chloro-1,3,5-triazine 12 (Scheme 1). In both cases, diisopropyle-
thylamine (DIEA) was used as a base. This reaction sequence could
be performed in the reverse order (i.e., 4-aminophenol with cya-
nuric chloride and then glycinamide hydrochloride) but was less
N
N
NH
N
N
N
NH
N
N
N
NH
N
OPO3H2
OPO3H2
OPO3H2
F
F
F
Me
N
H
N
H
N
HN
N
HN
N
HN
N
HN
N
HN
N
HN
N
N
N
Et
Et
N
N
N
N
N
N
N
N
N
N
Me
Cl
NH
NH
NH
HN
N
N
F
O
Cl
NH
10
5
6
7
8
9
Fig. 2. Patented structures 5–10.

246
C. Courme et al. / European Journal of Medicinal Chemistry 45 (2010) 244–255
OH
The latter was converted into the corresponding dibenzyl
phosphate 38 with in situ formed chlorodibenzylphosphite.
Debenzylation was performed by catalytic hydrogenation over Pd/C
to give aryl phosphoric acid 4 also of low solubility in usual
solvents, which was subsequently treated with sodium methoxide,
for the reason mentioned above, thus providing the desired sodium
salt 39.
Grb2-SH2 binding data for compounds 25, 26, and 39 were
determined using an ELISA assay [89] that measures their ability to
inhibit the binding of the phosphorylated peptide PSpYVNVQN
(Kd ¼ 9 nM) to the SH2 domain. Compounds 25 and 39 were found
to be inactive, whereas 26 binds to Grb2 with an IC50 value in the
OPO3H2
H2N
CONH2
NH2
2
H
N
1
N
CONH2
SNAr
Cl
N
Cl
HN
SNAr
N
N
N
N
Cl
O
Suzuki coupling
3
B(OH)2
2 meta
3 ortho
H2O3PO
high micromolar range [90] (IC50 ¼ 265
mM) (Fig. 4).
RO
Superimposition of the docked compounds 1, 25 and 26 shows
that, while the two phosphate groups occupy similar binding
positions, the carboxamide groups adopt different conformations
[91]. In agreement with the ELISA results, docking experiments by
molecular dynamics (MD) [92] predicted an improved pattern of
interactions for compound 26 compared to compound 25. Thus, the
carboxamide NH2 of 26 can form hydrogen bonds with the Lys 109
and Leu 120 residues in the specificity pocket (Fig. 5). For both 25
and 26, the phosphate group occupies the pTyr-binding pocket
with the expected polar interactions (Arg 86, Ser 88, Ser 90 and Ser
96). Neither 25 nor 26, however, gave rise to significant interactions
with Arg 67 while by contrast 1 closely interacts with it [91]. In the
second pTyr pocket, and for both compounds, the phosphate
interacts with Arg 142, Asn 143, and Ser 141.
We have then retained the scaffold of compound 26, and, with
the aid of MD simulations, we have attempted to evolve it to design
compounds fitting better into the Grb2 recognition site. Thus we
have considered in consecutive fashion compounds 40–44 (Fig. 6).
Thus 40, bearing both the malonic acid [17–22] and keto-acid
[93,94] groups as phosphate mimics, gives rise to more interactions
with Grb2 than 26. Indeed, the phenyl D-ring forms the desired
Fig. 3. Triazinyl-based bis-aryl phosphates 2 and 3 via trifunctionalization of cyanuric
chloride.
based extracellular Grb2-SH2 domain binding assays (vide infra).
Indeed, it was noted that only at concentrations less than 5%, did
DMSO not alter ELISA assays (i.e., no effect on peroxidase activity).
Water insoluble compounds 2 and 3 were subsequently trans-
formed into the water-soluble sodium salts 25 and 26 (resp.)
required for this test, since their phosphate groups bear a dianionic
charge at physiological pH [76].
We next turned our attention to the synthesis of 4-aryl-2-ani-
linopyrimidine 4. Among the traditional four synthetic routes
(A–D) [77–81] envisioned for the preparation of the target struc-
ture, we chose the Wendelin’s procedure involving the condensa-
tion of an aryl guanidine with aryloxyphenyl butenone 31 (Route C,
Scheme 5) [81]. This latter would be obtained from the Heck
coupling reaction between methylvinylketone and 1-(4-benzyloxy-
phenoxy)-3-bromobenzene. Interestingly, a recent report described
the mono-arylation of 2,4,6-trichloropyrimidine 33 via Suzuki
coupling, allowing possible access to 4 (Route E) [82].
cation–p interaction with Arg 67. In the pTyr pocket, one malonate
First, Heck coupling [83] of 14 with methylvinylketone (MVK),
according to the protocole of Xu et al. [84], furnished the required
aryloxyphenyl butenone 31 (Scheme 6). Note that the yield of the
coupling compound was low (45%) when 2 equivalents of MVK
were used, probably due to the polymerization of MVK [85].
The 2-arylaminopyrimidinyl-based phosphate 4 was prepared
as shown in Scheme 7. High-temperature condensation [86] of aryl
guanidine 34 [87] and aryloxyphenyl butenone 31 provided the
desired heterocyclic structure 35. It is noteworthy that under
carboxylate also binds to this residue by hydrogen bonding, and the
interactions that involved the phosphate group take place as well
with the malonate group. In the second pTyr pocket, however, the
keto functionality is not well oriented for binding [91]. To improve
its orientation in this pocket, we have next replaced the amino-
phenyl fragment of 40 by the aminopyrrole fragment of 41. While
this indeed leads to a flip of the keto group, no improvement in the
polar interactions takes place. The consecutive introduction of the
(R,S)-trans cyclopropane ring (42) instead of the phenyl C-ring
enables a better anchoring to the protein (Fig. 7). The malonate is
hydrogen-bonded to Arg 86, Ser 88, Ser 90, Ser 96 and Lys 109 in the
pTyr-binding pocket, the carboxamide group satisfies hydrogen
bonding requirements with Lys 109 and Leu 120 in the specificity
pocket. In the second phosphate pocket, polar interactions take
place between the carboxylate group and Arg 142, Ser 141, and Asn
milder conditions
– less equivalents of amidine and lower
temperature – the yield was much reduced. Subsequently, the
nitrile group of 35 was transformed into a tetrazole with trime-
thylsilyl azide in the presence of catalytic tetrabutylammonium
fluoride under solventless conditions [88]. The yield of 36 was only
25% at 85 ꢀC but increasing the temperature to 120 ꢀC afforded
a significantly enhanced 89% isolated yield. Hydrogenation of 36
over 10% Pd/C afforded phenol 37.
143. Cation–p interactions are also observed between Arg 142 and
the keto-acid-pyrrole junction. To the best of our knowledge, the
use of the keto-acid group as a phosphate mimic has not been yet
reported in the design of Grb2-SH2 inhibitors.
OH
Recently, Combs et al. [95] reported a structure-based design of
potent protein phosphatase 1B (PTP1B) inhibitors that incorporate
the novel 1,1-dioxido-isothiazolidin-3-one (IZD) pTyr mimetic
containing the highly delocalized anion of the cyclic N-acylsulfo-
namide heterocycle at physiological pH. Such a heterocycle is
unexplored for the search of inhibitors of the Grb2-SH2. It was
therefore adopted to replace the keto-acid group of 42 thus leading
H
N
H
N
Cl
N
Cl
Cl
N
CONH2
HN
N
CONH2
a
b
N
N
N
N
N
N
Cl
Cl
Cl
11
12
to 43. MD reveals the stabilizing role of this group via cation–
p
interactions but at the cost of the interaction of the carboxamide
[91]. As a further step, we have shifted the IZD group to the next
position into the pyrrole ring, giving compound 44. MD now shows
Scheme 1. Synthesis of triazine 12. Reagents and conditions: (a) Glycinamide hydro-
chloride (1 eq), DIEA (2.2 eq), THF, 0 ꢀC, 1.5 h, 74%; (b) 4-Aminophenol (1 eq), DIEA
(1 eq), THF, 0 ꢀC, 30 min then 25 ꢀC, 1 h, 87%.

C. Courme et al. / European Journal of Medicinal Chemistry 45 (2010) 244–255
247
Br
B(OH)2
RO
BnO
a (i)
F
c
Br
O
O
OH
OH
13 R = OH
14 R = Bn
15
b
(ii)
Br
B(OH)2
O
O
a (i)
F
c
Br
RO
BnO
(ii)
16 R = OH
17 R = Bn
18
b
Scheme 2. Preparation of the meta- and ortho-substituted boronic acid 15 and 18. Reagents and conditions: (a) (i) MeONa (1.1 eq), MeOH, 25 ꢀC, 3 h, (ii) 1-Bromo-3-fluorobenzene
(1 eq) or 1-bromo-2-fluorobenzene (1 eq), NMP, 180 ꢀC, 15 h, 13 (43%), 16 (16%); (b) K2CO3 (1.5 eq), BnBr (1.5 eq), DMF, 25 ꢀC, 2 h, 14 (84%), 17 (95%); (c) (i) 2 M n-BuLi in THF (1.2 eq),
ꢁ78 ꢀC, 45 min, (ii) B(OiPr)3 (2.5 eq), ꢁ78 ꢀC, 1 h then 25 ꢀC, 1 h, (iii) 1 M HCl, 0 ꢀC / 25 ꢀC, 1 h, THF, 15 (81%), 18 (74%).
that its three pharmacophores are involved in Grb2 binding, while
retaining the cation– interactions between, on the one hand, the
new opportunities in the design of non-peptidic, non-phosphate
inhibitors of Grb2-SH2.
p
phenyl D-ring and Arg 67, and, on the other hand, the IZD ring and
Arg 142 (Fig. 8).
4. Experimental section
3. Conclusion
4.1. General information
In summary, we have illustrated the syntheses of novel
compounds, i.e., the triazinyl-based bis-aryl phosphates 2 and 3,
and the 2-arylaminopyrimidyl-based aryl phosphate 4. Tri-
functionalization of cyanuric chloride allowed us to prepare 2 and
3. The key step was the Suzuki cross-coupling between the required
diaryl ether boronic acid (15, 18) and the 2,4-diamino-6-chloro-
1,3,5-triazine derivative 12. Bis-phosphorylation completed the
syntheses. The aminopyrimidine scaffold in 4 was prepared by
condensation of aryl guanidine 34 with aryloxyphenyl butenone 31,
this latter being obtained via a Heck reaction. Introduction of the
tetrazole and phosphate groups was then successively performed.
These designed compounds were evaluated for their ability to bind
to the SH2 domain of Grb2. Thus, the water-soluble sodium salt 26
of 3 was found to be a ligand for Grb2 in the high micromolar range
All commercial reagents were used without purification and all
solvents were reaction grade. When necessary, solvents were
previously dried over molecular sieves. Tetrahydrofuran was also
dried over molecular sieves 4 Å unless otherwise stated (distilled
from sodium/benzophenone under argon). All reactions were per-
formed under an inert atmosphere of argon unless otherwise
stated. All reaction mixtures were stirred magnetically and moni-
tored by thin-layer chromatography using Merck silica gel 60 F254
visualized with UV light. Flash chromatography were performed
using SDS silica gel 60 (35–70 m). Melting points (uncorrected)
,
m
were determined on a Kofler bench. Mass spectra were recorded on
a Nermag R10-10C MS (CI) or a ZQ 2000 MS (ES) or a MS Station
JMS-700 JEOL (FAB).
1H, 13C and 31P NMR spectra were recorded on a Bruker AM 300
spectrometer. Chemical shift values are reported in parts per
million (ppm) and coupling constants in Hertz (Hz). In order to
simplify the reading of the NMR spectra interpretations (COSY,
HMQC, HMBC), the following attributions were chosen (Fig. 9):
Letter ‘‘A’’ will always refer to the heterocyclic core – whether it is
a pyrimidine or a triazine – and letter ‘‘B’’ to the aryl group linked to
this heterocycle through an NH bridge. ‘‘C’’ and ‘‘D’’ will refer to the
diaryl ether and finally ‘‘E’’ will refer to the benzyl groups, either
benzyloxy or dibenzylphosphates. Two AA0BB0 systems were
(IC50 ¼ 265
mM). Subsequently, computer-aided step-wise modifi-
cations of its 1,3,5-trisubstituted triazine scaffold led to novel
potential ligands bearing exclusively phosphate mimics, as exem-
plified by compounds 42 and 44. Promising fragments in their
construction are a cyclopropane ring, a keto-acid group, and a 1,1-
dioxido-isothiazolidin-3-one, for which there are no precedents, to
our knowledge, in the design of Grb2-SH2 inhibitors. MD simula-
tions have predicted an enriched pattern of H-bonds and cation–
p
interactions with this protein. Such preliminary results could open
OH
OH
OH
H
N
H
N
H
15
18
HN
N
CONH2
HN
N
CONH2
HN
O
N
N
CONH2
N
N
N
N
N
N
a
a
RO
Cl
12
O
R'O
21 R' = Bn
22 R' = H
19 R = Bn
20 R = H
b
b
Scheme 3. Suzuki coupling. Reagents and conditions: (a) 15 (1.4 eq) or 18 (1.2 eq), Pd(PPh3)4 (0.02 eq), K2CO3 (3 eq), CH3CN/H2O (1:1), reflux, 6 h, 19 (67%), 21 (51%); (b) H2, Pd/C
10% (10% w/w), MeOH/AcOH (4:1), 25 ꢀC, 4 days, 20 (81%), 22 (80%).

248
C. Courme et al. / European Journal of Medicinal Chemistry 45 (2010) 244–255
OH
OPO(OBn)2
OPO(OR)2
H
N
H
H
N
HN
N
CONH2
HN
N
N
CONH2
HN
N
CONH2
a
b
N
N
N
N
N
N
O
O
O
HO
(BnO)2OPO
(RO)2OPO
meta 2 (R = H)
meta 25 (R = Na)
ortho 3 (R = H)
meta 20
ortho 22
meta 23
ortho 24
c
c
ortho 26 (R = Na)
Scheme 4. Reagents and conditions: (a) HPO(OBn)2 (5 eq), DIEA 8 eq, DMAP (1 eq), CH3CN/CCl4 5:1, ꢁ15 ꢀC, 3 h, 23 (79%) and 24 (69%) from 20 and 22, respectively; (b) H2, Pd/C 10%
(10% w/w), MeOH, 25 ꢀC, 5 days, 2 (55%), 3 (44%); (c) MeONa (4 eq), MeOH, 0 ꢀC, 15 min then 25 ꢀC, 1 h, 25 (99%), 26 (99%).
observed for the two para-substituted aromatic rings B and D.
Compounds 19, 23 and 35 are described below as examples.
(s, 1H, CH2), 4.02 (s, 1H, CH2), 6.72 (s, 1H, CONH2), 6.92 (s, 1H,
CONH2), 9.32 (s, 1H, NH); 13C NMR (75 MHz; THF-d8):
d 43.4 (CH2),
166.3 (2A), 169.6 (CONH2), 170.3 (4A, 6A); MS (ESþ) m/z: 222
[M þ H]þ, 244 [M þ Na]þ.
4.2. Experimental procedures
4.2.2. 2-[4-Chloro-6-(4-hydroxyphenylamino)-1,3,5-triazin-2-
ylamino]acetamide (12)
4.2.1. 2-(4,6-Dichloro-1,3,5-triazin-2-ylamino)-acetamide (11)
Diisopropylethylamine (700 mL, 4.01 mmol) was added to
Diisopropylethylamine (1 mL, 5.74 mmol) was added to a solu-
tion of 11 (1.29 g, 5.81 mmol) and 4-aminophenol (634 mg,
5.81 mmol) in dry THF (15 mL) at 0 ꢀC. The mixture was stirred at
0 ꢀC for 30 min and then at room temperature for 1 h. The precip-
itate was filtered, washed with THF and water and then dried over
P2O5, yielding to 36 as a strongly insoluble white powder (1.33 g,
77%). Furthermore, the filtrate was extracted with ethyl acetate and
a solution of cyanuric chloride (334 mg, 1.81 mmol) and glycina-
mide hydrochloride (200 mg,1.81 mmol) in dry THF (15 mL) at 0 ꢀC.
The mixture was stirred at 0 ꢀC for 1.5 h and the solvent removed
under reduced pressure. Purification by flash chromatography with
cyclohexane/ethyl acetate (gradient from 50:50 to 0:100) and then
methanol/ethyl acetate (5:95) furnished 11 as an off-white powder
(296 mg, 74%). Mp 188–190 ꢀC; 1H NMR (300 MHz; THF-d8):
d
4.01
O
N
N
NH
N
31
Cl
Cl
N
Ar
1. ArLi 29, ether, -30 to 0 °C
DMA, 160 °C
2. DDQ, ether-THF, 0 °C
Route A
N
N
Ar'NH2
Route C
NH
NH2
27
HN
N
Ar'NH
ethylene glycol
reflux
Route B
Route D
N
N
ArB(OH)2 30, Pd(PPh3)4
Me2N
Me
1 M Na2CO3 benzene, reflux
H2O3PO
O
Cl
32
O
Ar
2-methoxyethanol, reflux
28
4
ethylene glycol
reflux
BnO
R
Ar'NH2
Ar =
O
1. ArB(OH)2 30, Pd(OAc)2
PPh3, Na2CO3
Route E
DME/H2O, 70 °C
2. MeMgBr, THF, -78 °C
Ar' =
Cl
Cl
N
N
Cl
33
Scheme 5. Synthetic routes envisioned to the pyrimidinyl-based aryl phosphate 4. Route A (see refs [77–79]): Addition of aryloxyphenyllithium 29 to 2-chloro-4-methylpyrimidine
27 followed by DDQ oxidation. Route B (see refs [77] and [80]): Suzuki coupling of 2,4-dichloro-6-methylpyrimidine 28 with aryloxyphenyl boronic acid 30. Route C (see ref. [81]
and refs cited therein): Condensation of an aryl guanidine with aryloxyphenyl butenone 31. Route D (see ref. [77]): Condensation of an aryl guanidine with
butenone 32. Route E (see ref. [82]): Suzuki coupling of 2,4,6-trichloropyrimidine 33 with aryloxyphenyl boronic acid 30.
b-dimethylamino-

C. Courme et al. / European Journal of Medicinal Chemistry 45 (2010) 244–255
249
100
O
90
Br
80
BnO
BnO
a
70
PSpYVNVPN
O
60
50
40
30
20
10
0
O
14
31
Triazine 26
Scheme 6. Synthesis of the diaryl ether-substituted
a
-enone 31. (a) methylvinylketone
(4 eq), Pd(OAc)2 (0.1 eq), PPh3 (0.2 eq), NEt3 (8 eq), sealed tube, DMF, 150 ꢀC, 4 h, 83%.
the combined extracts were washed with water and brine, dried
over magnesium sulfate and evaporated under reduced pressure to
give an additional portion of 12 as a white powder (168 mg, overall
yield: 87%). Mp 256 ꢀC with decomposition (recryst. from ethanol/
-10.0-9.5 -9.0 -8.5 -8.0 -7.5 -7.0 -6.5 -6.0 -5.5 -5.0 -4.5 -4.0 -3.5 -3.0 -2.5 -2.0 -1.5 -1.0
log C
DMF); 1H NMR (300 MHz; DMSO-d6):
d 3.78 (s, 2H, CH2), 6.67–6.69
Fig. 4. Competitive ELISA assay (PSpYVNVPN as the reference peptide).
(m, 2H, 2B, 6B), 7.06 (br s, 1H, NH), 7.42–7.45 (m, 2H, 3B, 5B), 8.08 (br
s,1H, NH), 9.20 (br s,1H, NH), 9.84 (br s,1H, NH); 13C NMR (75 MHz;
DMSO-d6): d 43.5 (CH2), 115.1 (2B, 6B), 121.8 (3B, 5B), 130.3 (1B), 153.2
(4B), 163.1 (4A or 6A), 165.9 (2A or CONH2), 167.6 (4A or 6A), 170.6 (2A
or CONH2); MS (ESþ) m/z: 295 [M þ H]þ, 317 [M þ Na]þ MS (ESꢁ) m/
z: 293 [M ꢁ H]ꢁ.
pressure. After a purification by flash chromatography with cyclo-
hexane/ethyl acetate (gradient from 99:1 to 90:10), 4-arylox-
yphenol 13 was obtained as a colourless oil (1.02 g, 43%). 1H NMR
(300 MHz; CDCl3):
6.86–6.88 (m, 1H, 4C), 6.92–6.95 (m, 2H, 2D, 6D), 7.05–7.07 (m, 1H,
2C), 7.15–7.17 (m, 2H, 5C, 6C); 13C NMR (75 MHz; CDCl3):
116.1 (4C),
d 4.96 (s, 1H, OH), 6.82–6.86 (m, 2H, 3D, 5D),
4.2.3. 3-(4-Hydroxyphenoxy)-1-bromobenzene (13)
A 4.6 M solution of sodium methoxide (2.17 mL, 9.98 mmol) was
added to a solution of hydroquinone (1 g, 9.08 mmol) in dry
methanol (2 mL). The mixture was stirred at room temperature for
3 h and the methanol was removed under reduced pressure. Dry N-
methylpyrrolidinone (8 mL) and 1-bromo-3-fluorobenzene (1 mL,
8.92 mmol) were added and the mixture was heated at 180 ꢀC for
15 h. Water was then added and the mixture was extracted with
ether. The combined organic extracts were washed with water and
brine, dried over magnesium sulfate and evaporated under reduced
d
116.5 (3D, 5D), 120.5 (2C), 121.4 (2D, 6D), 122.8 (1C), 125.4 (6C), 130.7
(5C), 149.3 (1D), 152.2 (4D), 159.4 (3C); MS (ESꢁ) m/z: 263 [M ꢁ H]ꢁ,
265 [M ꢁ H]ꢁ.
4.2.4. 3-(4-Benzyloxyphenoxy)-1-bromobenzene (14)
Potassium carbonate (2.4 g, 17.4 mmol) was added to a solution
of 13 (3 g, 11.3 mmol) in dry DMF (50 mL). Benzyl bromide (2.1 mL,
17.6 mmol) was then added and the mixture was stirred at room
N
N
NH
N
CN
CN
CN
a
c
e
b
HN
N
HN
N
31
N
N
NH2
HN
NH2
NH
BnO
RO
O
O
34
35
36 R = Bn
37 R = H
d
N
N
NR
N
N
N
NH
N
f
HN
N
HN
N
N
N
R2O3PO
(BnO)2OPO
O
O
38
4
R = H
g
39 R = Na
Scheme 7. Synthesis of the 2-arylaminopyrimidinyl-based phosphate 4 and its sodium salt 39. Reagents and conditions: (a) (i) H2NCN (1 eq), HNO3 (1.6 eq), EtOH, reflux, 6 h, (ii)
NaOH (1 eq), H2O, reflux, 5 min, 15%; (b) 34 (3 eq) and 31 (1 eq), DMA, 160 ꢀC, 6 h, 81% based on compound 31; (c) Me3SiN3 (1.5 eq), Bu4NF.3H2O (0.5 eq), solventless, 120 ꢀC, 6 h,
89%; (d) H2, Pd/C 10% (10% w/w), MeOH/CH2Cl2 (1:1), 25 ꢀC, 24 h, 90%; (e) HPO(OBn)2 (5 eq), DIEA (7 eq), DMAP (0.5 eq), CH3CN/CCl4 (5:1), ꢁ15 ꢀC, 2.5 h, 61%; (f) H2, Pd/C 10% (10%
w/w), MeOH, 25 ꢀC, 10 days, 49% (g) MeONa (3 eq), MeOH, 0 ꢀC, 1 h, 100%.

250
C. Courme et al. / European Journal of Medicinal Chemistry 45 (2010) 244–255
Fig. 7. Predicted binding mode of compound 42. The amino acid residues involved in
interaction are Asn 143, Arg 142, Ser 141, Leu 120, Lys 109, Ser 96, Arg 86, Ser 88 and
Arg 67 (from left to right).
Fig. 5. Predicted binding mode of compound 26 to Grb2-SH2. The atoms of 26 are
colored with carbon in cyan, oxygen in red, nitrogen in blue, and phosphorus in
orange. Amino acid residues important for binding are labeled in grey. Intermolecular
hydrogen bonds are shown by yellow dashed lines. The amino acid residues involved
in interaction are Asn 143, Arg 142, Ser 141, Leu 120, Lys 109, Ser 96, Arg 86, Ser 88 and
Ser 90 (from left to right) This illustration was prepared by PyMOL (Delano Scientific,
San Carlos, CA).
evaporated under reduced pressure to give a white solid. Purifica-
tion by flash chromatography with cyclohexane/ethyl acetate
(gradient from 80:20 to 0:100) and then methanol/ethyl acetate
(5:95) afforded 15 as a white powder (1.81 g, 81%). Mp 162 ꢀC; 1H
NMR (300 MHz; CDCl3): d 5.06 (s, 2H, CH2), 6.98–7.01 (m, 4H, 2D, 3D,
5D, 6D), 7.15 (dd, Jo ¼ 8.0 Hz, Jm ¼ 1.7 Hz, 1H, 4C), 7.34–7.46 (m, 6H,
temperature for 2 h. Water was added and the mixture was
extracted with ether. The combined extracts were washed with
water and brine, dried over magnesium sulfate and evaporated
under reduced pressure. Purification by flash chromatography with
cyclohexane/ethyl acetate (100:1) gave 14 as a white powder
(3.39 g, 84%). Mp 96 ꢀC (recryst. from ethyl acetate/pentane). 1H
2E, 3E, 4E, 5E, 6E, 5C), 7.86 (m, 1H, 2C), 7.89–7.91 (m, 1H, 6C); 13C NMR
(75 MHz; CDCl3):
d 70.6 (CH2), 116.0 (3D, 5D), 120.5 (2D, 6D), 122.2
(4C), 125.1 (2C), 127.6 (2E, 6E), 128.0 (4E), 128.7 (3E, 5E), 129.5 (5C),
130.0 (6C), 131.9 (1C), 137.0 (1E), 150.7 (1D), 155.1 (4D), 158.0 (3C); MS
(CI) m/z: 321 [M þ H]þ, 338 [M þ NH4]þ.
NMR (300 MHz; CDCl3):
d 5.06 (s, 2H, CH2), 6.87–6.91 (m, 1H, 4C),
4.2.6. 2-(4-Hydroxyphenoxy)-1-bromobenzene (16)
6.99 (s, 4H, 2D, 3D, 5D, 6D), 7.08–7.09 (m,1H, 2C), 7.15–7.17 (m, 2H, 5C,
6C), 7.34–7.47 (m, 5H, 2E, 3E, 4E, 5E, 6E); 13C NMR (75 MHz; CDCl3):
Compound 16 was prepared as 13, from hydroquinone (12 g,
109 mmol) and 1-bromo-2-fluorobenzene (12 mL, 110 mmol).
Purification by flash chromatography with cyclohexane/ethyl
acetate (gradient from 98:2 to 90:10) gave 16 as a white powder
(4.6 g, 16%). Mp 85 ꢀC (recryst. from ethyl acetate/pentane); 1H
d
70.4 (CH2), 116.1 (3D, 5D), 116.2 (4C), 120.5 (2C), 121.2 (2D, 6D), 122.8
(1C), 125.4 (6C), 127.5 (2E, 6E), 128.1 (4E), 128.6 (3E, 5E), 130.7 (5C),
136.9 (1E), 149.5 (1D), 155.6 (4D), 159.4 (3C). Anal. Calcd for
C19H15BrO2: C, 64.24; H, 4.26. Found: C, 64.17; H, 4.37.
NMR (300 MHz; CDCl3): d 4.92 (br s, 1H, OH), 6.80–6.84 (m, 3H, 3D,
5D, 3C), 6.88–6.93 (m, 2H, 2D, 6D), 6.85 (dd, Jo ¼ 7.9 Hz, Jm ¼ 1.5 Hz,
1H, 5C), 7.21 (ddd, Jo ¼ 8.1 Hz, Jo ¼ 7.9 Hz, Jm ¼ 1.5 Hz, 1H, 4C), 7.60
(dd, Jo ¼ 7.8 Hz, Jm ¼ 1.5 Hz, 1H, 6C); 13C NMR (75 MHz; CDCl3):
4.2.5. 3-(4-Benzyloxyphenoxy)phenylboronic acid (15)
A solution of 2 M n-butyllithium in THF (3.8 mL, 7.8 mmol) was
added dropwise to a solution of compound 14 (2.49 g, 7.01 mmol)
in dry THF (Na/benzophenone) (25 mL) at ꢁ78 ꢀC. The resulting
mixture was stirred at ꢁ78 ꢀC for 45 min and triisopropyl borate
(4.0 mL, 17.3 mmol) was added. The mixture was again stirred at
ꢁ78 ꢀC for 1 h and then at room temperature for a further 1 h. Ether
(25 mL) and 1 M HCl (25 mL) were added at 0 ꢀC. The mixture was
stirred at 0 ꢀC for 10 min, at room temperature for 1 h and then the
mixture was extracted with ether. The combined extracts were
washed with water and brine, dried over magnesium sulfate and
d
113.7 (1C), 116.4 (3D, 5D), 118.8 (3C), 120.4 (2D, 6D), 124.2 (5C), 128.5
(4C), 133.7 (6C), 150.1 (1D), 151.8 (4D), 154.8 (2C); MS (ESꢁ) m/z: 263
[M ꢁ H]ꢁ; Anal. Calcd for C12H9BrO2: C, 54.27; H, 3.42. Found C,
54.57; H, 3.39.
4.2.7. 2-(4-Benzyloxyphenoxy)-1-bromobenzene (17)
Potassium carbonate (3.2 g, 23.1 mmol) was added to a solution
of 16 (4.09 g, 15.4 mmol) in dry DMF (50 mL). Benzyl bromide
O
H
O
O
N
O
O
CO2H
CO2H
HN
S
CO2H
S
O
O
O
O
HN
HN
HN
CONH2 HN
HN
H
N
H
N
H
N
H
N
H
N
HN
N
CONH2
HN
N
CONH2
HN
N
CONH2 HN
N
N
CONH2
N
N
N
N
N
O
N
N
O
N
N
O
N
3
R
O
O
S
HO2C
HO2C
CO2H
CO2H
HO2C
HO2C
HO2C
40
41
42
43
44
CO2H
CO2H
CO2H
Fig. 6. De novo design from the heterocycle scaffold of 3: generation of compounds 40–44.

C. Courme et al. / European Journal of Medicinal Chemistry 45 (2010) 244–255
251
164.3 (2C); MS (ESꢁ) m/z: 319 [M ꢁ H]ꢁ; Anal. Calcd for C19H17BO4:
C, 71.28; H, 5.35. Found: C, 70.59; H, 5.36.
4.2.9. 2-[4-[3-(4-Benzyloxyphenoxy)-phenyl]-6-(4-
hydroxyphenylamino)-1,3,5-triazin-2-ylamino]-acetamide (19)
Triazine 12 (304 mg, 1.03 mmol), boronic acid 15 (463 mg,
1.45 mmol) and potassium carbonate (409 mg, 3.09 mmol) were
placed into an acetonitrile/water mixture (30 mL, 1:1). The solvent
was then degassed as follows: the mixture was frozen in liquid
nitrogen, placed under high vacuum for 5 min to remove dissolved
gases, and then purged with argon. The operation was reiterated
twice before addition of palladium tetrakis(triphenylphosphine)
(24 mg, 0.021 mmol). Themixturewas then refluxed for 6 h, filtrated
upon celite to remove the catalyst and evaporated under reduced
pressure to give a brown solid. Purification by flash chromatography
with ethyl acetate/methanol(gradient from100:0to92:8)gave19 as
a beige powder (370 mg, 67%). Mp 101–103 ꢀC; 1H NMR (300 MHz;
Fig. 8. Predicted binding mode of compound 44. The amino acid residues involved in
interaction are Arg 142, Ser 141, Lys 109, Ser 96, Arg 86, Ser 88, Arg 67 and Ser 90.
MeOD): d 4.02 (s, 2H, CH2–CONH2), 5.06 (s, 2H, CH2–Ph), 6.72–6.75
(m, 2H, 3B, 5B), 6.99–7.00 (m, 4H, 2D, 3D, 5D, 6D), 7.09–7.11 (m,1H, 4C),
7.27–7.46 (m, 8H, 2E, 3E, 4E, 5E, 6E, 2B, 6B, 5C), 7.89 (s,1H, 2C), 8.04 (brd,
Jo ¼ 7.4 Hz,1H, 6C); 13C NMR (75 MHz; MeOD):
d 44.9 (CH2–CONH2),
(2.7 mL, 23.1 mmol) was then added and the mixture was stirred at
room temperature for 2 h. Water was added and the mixture was
extracted with ether. The combined extracts were washed with
water and brine, dried over magnesium sulfate and evaporated
under reduced pressure to give an off-white solid. Purification by
flash chromatography with cyclohexane/ethyl acetate (100:0.5)
furnished 17 as a white powder (5.18 g, 95%). Mp 71 ꢀC (recryst.
71.5 (CH2–Ph),116.1 (3B, 5B),117.1 (3D, 5D),118.2 (2C),121.9(2D, 6D, 4C),
123.3 (2B, 6B),123.5 (6C),127.3 (5C),128.7 (2E, 6E),128.9 (4E),129.5 (3E,
5E), 132.6 (1B), 138.7 (1E), 140.1 (1C), 151.5 (1D), 154.3 (4B), 156.7 (4D),
160.0 (3C),165.9 (4A or 6A),167.8 (2A),171.7 (4A or 6A),176.0 (CONH2);
MS (CI) m/z: 535 [M þ H]þ.
from ethyl acetate/pentane); 1H NMR (300 MHz; CDCl3):
2H, CH2), 6.89 (dd, Jo ¼ 8.1 Hz, Jm ¼ 1.4 Hz, 1H, 3C), 6.96–7.00 (m, 5H,
5C, 2D, 3D, 5D, 6D), 7.23 (ddd, Jo ¼ 7.4 Hz,
¼ 8.1 Hz, Jo
d 5.07 (s,
4.2.10. 2-(4-(3-(4-Hydroxyphenoxy)phenyl)-6-(4-
hydroxyphenylamino)-1,3,5-triazin-2-ylamino)-acetamide (20)
10% Palladium on carbon (24 mg), was added to a solution of 19
(242 mg, 0.453 mmol) in methanol/acetic acid (25 mL, 4:1). The
mixture was stirred under H2 at room temperature for 4 days, fil-
trated upon celite to remove the catalyst and evaporated under
reduced pressure to give a yellow oil. Purification by flash chroma-
tography with a gradient from ethyl acetate/cyclohexane (75:25) to
ethyl acetate/methanol (95:5) gave 20 as white solid (162 mg, 80%).
4–3
4–5
Jm ¼ 1.5 Hz, 1H, 4C), 7.36–7.48 (m, 5H, 2E, 3E, 4E, 5E, 6E), 7.64 (dd,
Jo ¼ 7.9 Hz, Jm ¼ 1.5 Hz, 1H, 6C); 13C NMR (75 MHz; CDCl3):
d 70.6
(CH2), 113.9 (1C), 116.0 (3D, 5D), 119.0 (3C), 120.2 (2D, 6D), 124.2 (5C),
127.5 (2E, 6E), 128.1 (4E), 128.4 (4C), 128.6 (3E, 5E), 133.4 (6C), 137.0
(1E), 150.3 (1D), 154.9 (2C), 155.3 (4D); MS (ESþ) m/z: 377 [M þ Na]þ,
393 [M þ K]þ; Anal. Calcd for C19H15BrO2: C, 64.24; H, 4.26. Found:
C, 64.14; H, 4.23.
Mp 236–238 ꢀC; 1H NMR (300 MHz; MeOD):
d 4.02 (s, 2H, CH2–
CONH2), 6.72–6.75 (m, 2H, 3B, 5B), 6.79–6.82 (m, 2H, 3D, 5D), 6.88–
6.91 (m, 2H, 2D, 6D), 7.06 (d, Jo ¼ 8.4 Hz,1H, 4C), 7.33–7.37 (m,1H, 5C),
7.42–7.45 (m, 2H, 2B, 6B), 7.87 (s, 1H, 2C), 8.02 (d, Jo ¼ 7.5 Hz, 1H, 6C);
4.2.8. 2-(4-Benzyloxyphenoxy)phenylboronic acid (18)
Compound 18 was prepared as 15, i.e. from 17 (4.4 g, 12.4 mmol)
instead of 14. Purification by flash chromatography with cyclo-
hexane/ethyl acetate (2:1) afforded 18 as a white powder (2.94 g,
74%). Mp 100 ꢀC (recryst. from ethyl acetate/pentane); 1H NMR
13C NMR (300 MHz; MeOD):
d 44.9 (CH2–CONH2),116.2 (3B, 5B),117.3
(3D, 5D, 2C), 121.5 (4C), 122.1 (2D, 6D), 123.3 (2B, 6B, 6C), 130.4 (5C),
132.6 (1B), 140.0 (1C), 150.3 (1D), 154.3 (4B), 155.0 (4D), 160.4 (3C),
165.8 (4A or 6A), 167.8 (2A), 171.7 (4A or 6A), 175.9 (CONH2); MS (ESꢁ)
m/z: 443 [M ꢁ H]ꢁ; MS (ESþ) m/z: 445 [M þ H]þ.
(300 MHz; CDCl3):
d
5.08 (s, 2H, CH2), 6.67 (d, Jo ¼ 8.3 Hz, 1H, 3C),
7.02 (m, 4H 2D, 3D, 5D, 6D), 7.09 (ddd, Jo ¼ 7.3 Hz, Jo ¼ 7.3 Hz,
Jm ¼ 0.8 Hz, 1H, 5C), 7.33 (m, Jo ¼ 7.3 Hz, Jm ¼ 1.7 Hz, 1H, 4C), 7.35–
7.47 (m, 5H, 2E, 3E, 4E, 5E, 6E), 7.92 (dd, Jo ¼ 7.3 Hz, Jm ¼ 1.7 Hz, 1H,
4.2.11. 2-(4-(2-(4-Benzyloxyphenoxy)phenyl)-6-(4-
6C); 13C NMR (75 MHz; CDCl3):
d 70.5 (CH2),116.1 (3C),116.1 (3D, 5D),
hydroxyphenylamino)-1,3,5-triazin-2-ylamino)-acetamide (21)
Compound 21 was prepared as 19, from triazine 12 (54 mg,
0.183 mmol) and boronic acid 18 (70 mg, 0.219 mmol). Purification
121.9 (2D, 6D), 122.6 (5C), 127.5 (2E, 6E), 128.1 (4E), 128.7 (3E, 5E),
132.7 (4C), 136.8 (1C), 136.8, 136.9 (6C, 1E), 148.6 (1D), 156.0 (4D),
O
P
BnO
5
2
1
3
E
O
OH
O
CN
4
4
4
4
6
5
6
3
2
3
2
5
6
3
2
5
B
B
B
6
1
1
HN
1
H
N
H
N
HN
N
A
CONH2
HN
N
A
CONH2
N
A
6
2
6
2
2
6
5
3
E
6
3
E
6
N
N
N
N
N
4
5
2
1
4
5
2
1
4
4
4
3
D
6
3
D
6
3
D
6
1
1
1
O
(BnO)2OPO
O
2
3
6
5
2
3
6
5
2
3
6
5
2
2
1
2
1
4
5
4
4
C
C
C
1
5
5
O
O
O
4
4
4
23
19
35
Fig. 9.

252
C. Courme et al. / European Journal of Medicinal Chemistry 45 (2010) 244–255
by flash chromatography with ethyl acetate afforded 21 as a beige
153.9 (2C), 157.1 (4D), 164.1 (6A), 165.4 (2A), 171.2 (4A), 171.6 (CONH2);
powder (50 mg, 51%). Mp 115–118 ꢀC; 1H NMR (300 MHz; MeOD):
MS (ESþ) m/z: 965 [M þ H]þ, 987 [M þ Na]þ.
d
3.95 (s, 2H, CH2–CONH2), 4.99 (s, 2H, CH2–Ph), 6.68 (br s, 2H, 3B,
5B), 6.86–6.92 (m, 5H, 2D, 3D, 5D, 6D, 3C), 7.14 (m, 1H, 5C), 7.27–7.41
4.2.15. 2-(4-(3-(4-Phosphoryloxyphenoxy)phenyl)-6-(4-
(m, 8H, 2E, 3E, 4E, 5E, 6E, 2B, 6B, 4C), 7.72–7.74 (m, 1H, 6C); 13C NMR
phosphoryloxyphenylamino)-1,3,5-triazin-2-ylamino)acetamide (2)
10% Palladium on carbon (9 mg) was added to a solution of
compound 23 (90 mg, 0.093 mmol) in methanol (10 mL), and the
mixture was stirred under H2 for 3 days at room temperature. The
catalyst was filtered using two filter papers that were washed with
methanol and THF – giving only impurities – and then with warm
water, for the solubility of compound 2 was very low. The latter
filtrate was then evaporated under reduced pressure to give a white
solid (41 mg, 73%). A portion of this product (25 mg) was purified
(75 MHz; MeOD): d 44.8 (CH2–CONH2), 71.5 (CH2–Ph), 116.1 (3B, 5B),
117.0 (3D, 5D), 119.6 (3C), 121.5 (2D, 6D), 123.7 (2B, 6B, 5C), 128.6 (2E,
6E), 128.9 (4E), 129.5 (3E, 5E), 130.4 (1C), 132.1 (1B, 6C), 132.5 (4C),
138.7 (1E), 152.1 (1D), 154.6 (4B), 156.4 (4D), 157.6 (2C), 165.1 (6A),
167.0 (2A), 173.0 (4A), 175.4 (CONH2); MS (ESþ) m/z: 535 [M þ H]þ,
557 [M þ Na]þ; MS (ESꢁ) m/z: 533 [M ꢁ H]ꢁ.
4.2.12. 2-(4-(2-(4-Hydroxyphenoxy)phenyl)-6-(4-
hydroxyphenylamino)-1,3,5-triazin-2-ylamino)-acetamide (22)
10% Palladium on carbon (22 mg), was added to a solution of 21
(223 mg, 0.417 mmol) in acetic acid (22 mL). The mixture was
stirred under H2 at room temperature for 48 h, filtrated upon celite
to remove the catalyst and evaporated under reduced pressure to
give a yellow oil. Purification by flash chromatography with ethyl
acetate/methanol (gradient from 99:1 to 95:5) gave 22 as a white
via semi-preparative HPLC (X-Terra C18 MS 5
m
m silica,150 ꢂ 19 mm
column, NH3 aq buffer pH 9/CH3CN, gradient from 90:10 to 50:50)
and compound 2 was obtained as a wþhite solid (19 mg, 55%).
Mp > 250 ꢀC; MS (ESþ) m/z: 605 [M þ H] , 627 [M þ Na]þ; HRMS
(ESꢁ) m/z: calcd for C23H22N6O10P2 603.0794 [M ꢁ H]ꢁ; found
603.0821.
solid (149 mg, 80%). Mp 148 ꢀC; 1H NMR (300 MHz; MeOD):
d
3.95
4.2.16. 2-(4-(2-(4-Phosphoryloxyphenoxy)phenyl)-6-(4-
(s, 2H, CH2–CONH2), 6.69–6.77 (m, 4H, 3B, 5B, 3D, 5D), 6.83–6.89
(m, 3H, 2D, 6D, 3C), 7.11 (br t, Jo ¼ 7.4 Hz,1H, 5C), 7.32–7.41 (m, 3H, 2B,
phosphoryloxyphenylamino)-1,3,5-triazin-2-ylamino)acetamide (3)
Compound 3 was prepared as compound 2, from 24 (87 mg,
0.090 mmol) and 10% palladium on carbon (9 mg). The aqueous
filtrate was evaporated under reduced pressure to give 3 as a white
solid (40 mg, 74%). A portion of this product (25 mg) was purified
6B, 4C), 7.70–7.74 (m, 1H, 6C); 13C NMR (75 MHz; MeOD):
d 44.8
(CH2–CONH2), 116.1 (3B, 5B), 117.1 (3D, 5D), 118.9 (3C), 121.7 (2D, 6D),
123.4 (5C), 123.7 (2B, 6B), 130.0 (1C), 132.1 (1B, 6C), 132.5 (4C), 150.5
(1D), 154.6 (4B), 156.8 (4D), 158.1 (2C), 165.0 (6A), 167.0 (2A), 173.1
(4A), 175.6 (CONH2); MS (ESþ) m/z: 445 [M þ H]þ; MS (ESꢁ) m/z:
443 [M ꢁ H]ꢁ.
via semi-preparative HPLC (X-Terra C18 MS 5
m
m silica,150 ꢂ 19 mm
column, NH3 aq buffer pH 9/CH3CN, gradient from 90:10 to 50:50)
and compound 3 was obtained as aꢁwhite solid (15 mg, 44%). Mp
238–240 ꢀC (decomposition); MS (ES ) m/z: 523 [M ꢁ PO3H2]ꢁ, 603
[M ꢁ H]ꢁ, 625 [M þ Na]ꢁ.
4.2.13. 2-[4-(3-(4-Dibenzylphosphoryloxyphenoxy)phenyl)-6-(4-
dibenzylphosphoryloxyphenylamino)-1,3,5-triazin-2-
ylamino]acetamide (23)
4.2.17. Sodium 2-(4-(3-(4-phosphonatooxyphenoxy)phenyl)-6-(4-
phosphonatooxyphenylamino)-1,3,5-triazin-2-ylamino)acetamide
(25)
Compound 23 was prepared from 20 (84 mg, 0.189 mmol), dii-
sopropylethylamine (265
mL, 1.52 mmol), N,N-dimethylaminopyr-
imidine (23 mg, 0.188 mmol) and dibenzyl phosphite (210
m
L,
Sodium methoxide (6.8 mg, 0.126 mmol) was added to
a suspension of compound 2 (19 mg, 0.031 mmol) in dry methanol
(8 mL) at 0 ꢀC. The mixture was stirred at 0 ꢀC for 15 min and then
at room temperature for 1 h. Methanol was removed under
reduced pressure to afford 25 as a water-soluble white powder
0.951 mmol). Purification by flash chromatography with ethyl
acetate afforded 23 as a colourless oil (144 mg, 79%). 1H NMR
(300 MHz; CDCl3):
d 3.99 (s, 2H, CH2–CONH2), 5.09–5.14 (m, 8H,
4 ꢂ O–CH2–C6H5), 6.96–7.08 (m, 6H, 2D, 3D, 5D, 6D, 3B, 5B), 7.30–7.32
(m, 21H, 4 ꢂ 2E, 4 ꢂ 3E, 4 ꢂ 4E, 4 ꢂ 5E, 4 ꢂ 6E, 5C), 7.50–7.53 (m, 2H,
2B, 6B), 7.74 (m, 1H, 4C), 7.90 (s, 1H, 2C), 8.06 (m, 1H, 6C); 13C NMR
(21.5 mg, 99%). 1H NMR (300 MHz; D2O):
d 4.05 (s, 2H, CH2–
CONH2), 7.08–7.11 (m, 2H, 3D, 5D), 7.24 (m, 5H, 2D, 6D, 3B, 5B, 4C),
(75 MHz; CDCl3):
d 44.3 (CH2–CONH2), 69.9 (O–CH2–C6H5), 70.0
7.48 (m, 3H, 2B, 6B, 5C), 7.75 (s, 1H, 2C), 7.89–7.90 (m, 1H, 6C); 13C
(O–CH2–C6H5),117.0 (2C),118.2 (4C),120.1,120.3,121.0 (3D, 5D, 3B, 5B,
2D, 6D), 121.5 (2B, 6B), 123.2 (6C), 128.0 (2E, 6E), 128.6 (4E), 128.8 (3E,
5E), 129.8 (5C), 135.4 (1E), 135.9 (1B), 138.4 (1C), 145.9 (4B), 146.2 (1D),
153.6 (4D), 157.5 (3C), 164.4, 166.1 (2A, 6A),170.6 (4A), 172.3 (CONH2);
MS (ESþ) m/z: 987 [M þ Na]þ; MS (ESꢁ) m/z: 963 [M ꢁ H]ꢁ.
NMR (75 MHz; D2O): d 43.4 (CH2–CONH2), 117.0 (2C), 120.5 (3D, 5D,
3B, 5B), 121.2 (4C), 121.7 (2D, 6D), 122.8 (2B, 6B, 6C), 130.0 (5C), 132.0
(1B), 137.2 (1C), 150.3, 150.7 (1D, 4D, 4B), 157.9 (3C), 161.2 (6A), 165.8
(2A), 171.1 (4A), 175.5 (CONH2); 31P NMR (121 MHz; D2O):
d 0.67; MS
(ESþ) m/z: 693 [M þ H]þ, 714 [M þ Na]þ, 737 [M þ 2Na]þ.
4.2.14. 2-(4-(2-(4-Dibenzylphosphoryloxyphenoxy)phenyl)-6-(4-
dibenzylphosphoryloxyphenylamino)-1,3,5-triazin-2-
ylamino)acetamide (24)
4.2.18. Sodium 2-(4-(2-(4-phosphonatooxyphenoxy)phenyl)-6-(4-
phosphonatooxyphenylamino)-1,3,5-triazin-2-ylamino)acetamide
(26)
Compound 24 was prepared as 23, from 22 (126 mg, 0.284 mmol),
Compound 26 was prepared as 25, from 3 (15 mg, 0.025 mmol)
and sodium methoxide (5.4 mg, 0.100 mmol). 26 was obtained as
a water-soluble white powder (17 mg, 99%). 1H NMR (300 MHz;
diisopropylethylamine (300
mL, 1.72 mmol), N,N-dimethylamino-
pyrimidine (35 mg, 0.287 mmol) and dibenzyl phosphite (250
mL,
1.13 mmol). Purification by flash chromatography with ethyl acetate
D2O): d 3.74 (s, 2H, CH2–CONH2), 6.75–6.81 (m, 2H, 3D, 5D), 6.94–
afforded 24 as a colourless oil (189 mg, 69%). 1H NMR (300 MHz;
6.97 (m, 3H, 2D, 6D, 3C), 6.99–7.02 (m, 2H, 3B, 5B), 7.10–7.15 (m, 1H,
5C), 7.22–7.25 (m, 2H, 2B, 6B), 7.34–7.39 (m, 1H, 4C), 7.48–7.51 (m,
CDCl3):
d
3.43 (s, 2H, CH2–CONH2), 5.09 (s, 4H, 2 ꢂ O–CH2–C6H5), 5.12
(s, 4H, 2 ꢂ O–CH2–C6H5), 6.84–6.86 (m, 2H, 3D, 5D), 7.01–7.08 (m, 5H,
2D, 6D, 3B, 5B, 5C), 7.31–7.35 (m, 21H, 4 ꢂ 2E, 4 ꢂ 3E, 4 ꢂ 4E, 4 ꢂ 5E,
4 ꢂ 6E, 3C), 7.48–7.54 (m, 3H, 2B, 6B, 4C), 7.94–7.96 (m, 1H, 6C); 13C
1H, 6C); 13C NMR (75 MHz; D2O):
d 43.3 (CH2–CONH2), 118.5 (3C),
119.3 (3D, 5D), 120.5 (3B, 5B), 121.4 (2D, 6D), 123.4 (2B, 6B), 123.7 (5C),
127.0 (1C), 130.5 (6C), 130.7 (1B), 131.7 (4C), 149.7 (1D), 150.7 (4B),
151.6 (4D), 154.8 (2C), 161.6 (6A), 165.0 (2A), 171.9 (4A), 174.8
NMR (75 MHz; CDCl3): d 42.7 (CH2–CONH2), 69.9 (O–CH2–C6H5), 70.0
(O–CH2–C6H5), 117.7 (3D, 5D), 118.8 (3C), 120.4 (3B, 5B), 121.0 (2D, 6D),
121.4 (2B, 6B),122.9 (5C),125.1 (1C),128.0 (2E, 6E),128.7 (4E),128.9 (3E,
5E), 131.5 (6C), 132.0 (4C), 135.4, 135.5 (1E, 1B), 144.5 (1D), 146.1 (4B),
(CONH2); 31P NMR (121 MHz; D2O): 0.60. MS (ESþ) m/z: 693
d
[M þ H]þ, 714 [M þ Na]þ, 737 [M þ 2Na]þ; HRMS (ESþ) m/z: calcd
for C23H18N6Na4O10P2 693.0228 [M þ H]þ; found 693.0215.

C. Courme et al. / European Journal of Medicinal Chemistry 45 (2010) 244–255
253
4.2.19. (E)-4-(3-(4-Benzyloxyphenoxy)phenyl)but-3-en-2-one (31)
Compound 14 (200 mg, 0.563 mmol), but-2-ene-3-one (192
2.31 mol), triethylamine (570 L, 4.9 mmol), palladium acetate
4.2.22. N-[4-(2H-Tetrazol-5-yl)phenyl]-4-[3-(4-
benzyloxyphenoxy)phenyl]-6-methylpyrimidin-2-amine (36)
Compound 35 (40 mg, 0.083 mmol), trimethylsilyl azide (16 mL,
mL,
m
(13 mg, 0.058 mmol), triphenylphosphine (30 mg, 0.112 mmol) and
dry DMF (2 mL) were placed in a sealed Pyrex tube. The mixture
was heated at 150 ꢀC for 4 h and cooled to room temperature before
safely opening the tube. Water was added and the mixture was
extracted with ether. The combined extracts were washed with
water and brine, dried over magnesium sulfate and evaporated
under reduced pressure to give a yellow oil. Purification by flash
chromatography with cyclohexane/ethyl acetate (3:1) furnished 31
as a yellow powder (161 mg, 83%). Mp 119 ꢀC (recryst. from
0.125 mmol) and tetrabutylammonium fluoride trihydrate (13 mg,
0.041 mmol) were introduced into a Pyrex tube. The tube was
sealed, heated at 120 ꢀC for 6 h and then cooled to room temper-
ature. Water was added and the mixture was extracted with
dichloromethane. The combined extracts were washed with water
and brine, dried over magnesium sulfate and evaporated under
reduced pressure to give 36 as a white powder (39 mg, 89%). Mp
129–131 ꢀC; 1H NMR (300 MHz; THF-d8):
d 2.45 (s, 3H, CH3), 5.06 (s,
2H, CH2), 7.03 (s, 4H, 2D, 3D, 5D, 6D), 7.05–7.06 (m,1H, 4C), 7.22 (s,1H,
5A), 7.25–7.34 (m, 3H, 3E, 4E, 5E), 7.38–7.41 (m, 3H, 2E, 6E, 5C), 7.80–
7.82 (m, 2H, 2C, 6C), 7.95–7.98 (m, 2H, 3B, 5B), 8.02–8.05 (m, 2H, 2B,
dichloromethane); 1H NMR (300 MHz; CDCl3):
d 2.36 (s, 3H, CH3),
5.07 (s, 2H, CH2), 6.63 (d, Jtrans ¼ 16.3 Hz, 1H, HC]CH–CO), 6.99 (s,
5H, 4C, 2D, 3D, 5D, 6D), 7.08 (s, 1H, 2C), 7.22–7.25 (m, 1H, 6C), 7.33–
7.44 (m, 7H, 5C, HC]CH–CO, 2E, 3E, 4E, 5E, 6E); 13C NMR (75 MHz;
6B), 9.13 (s, 1H, NH); 13C NMR (75 MHz; THF-d8):
d 23.3 (CH3), 69.9
(CH2), 107.7 (5A), 115.8 (3D, 5D), 116.1 (2C), 117.6 (4B), 118.4 (2B, 6B),
119.4 (4C), 120.5 (2D, 6D), 120.9 (6C), 127.2, 127.3, 127.4 (2E, 4E, 6E, 3B,
5B), 128.1 (3E, 5E), 129.7 (5C), 137.5 (1E), 139.3 (1C), 143.6 (1B), 146.2
(1D), 155.5 (4D), 156.4 (CN4H), 159.2 (3C), 160.2 (2A), 163.7 (4A), 168.7
(6A); MS (CI) m/z: 528 [M þ H]þ, 550 [M þ Na]þ.
CDCl3): d 27.6 (CH3), 70.6 (CH2), 116.2 (3D, 5D), 116.6 (2C), 119.8 (4C),
121.2 (2D, 6D), 122.7 (6C), 127.7 (2E, 6E), 127.8 (HC]CH–CO), 128.2
(4E), 128.8 (3E, 5E), 130.3 (5C), 136.2 (1C), 137.0 (1E), 143.0 (HC]CH–
CO), 149.8 (1D), 155.6 (4D), 159.2 (3C), 198.5 (CO); MS (CI) m/z: 345
[M þ H]þ; Anal. Calcd for C23H20O3: C, 80.21; H, 5.85. Found: C,
79.52; H, 5.86.
4.2.23. 4-(3-{6-Methyl-2-[4-(2H-tetrazol-5-yl)-phenylamino]-
pyrimidin-4-yl}-phenoxy)-phenol (37)
4.2.20. 1-(4-Cyanophenyl)guanidine (34)
10% Palladium on carbon (10 mg) was added to a solution of 36
in methanol/dichloromethane (3 mL, 1:1). The mixture was stirred
under H2 at room temperature for 24 h, filtrated upon celite to
remove the catalyst and evaporated under reduced pressure to give
Aryl guanidine 34 was prepared as described by Bauer and Saffir
[87]. 8 N nitric acid (5 mL, 41 mmol) was slowly added to a solution
of 4-aminobenzonitrile (3 g, 25 mmol) and cyanamide (1.08 g,
25 mmol) in ethanol (13 mL) under air atmosphere. The mixture
was then refluxed for 6 h. The guanidine nitrate that precipitated
upon cooling was filtered and washed, giving 1.16 g of a pale orange
powder (5.2 mmol, 21%). To a solution of this nitrate derivative in
refluxing water (10 mL) was added dropwise a 1 M sodium
hydroxide solution (5.2 mL, 5.2 mmol). The mixture was refluxed
for 5 min and the precipitate obtained upon cooling was filtered,
washed with water and dried to give 34 as a pale brown powder
(609 mg, 15% overall yield). Mp 220–222 ꢀC (recryst. from ethanol);
a
yellow solid. Purification by flash chromatography with
dichloromethane/methanol (93:7) gave 37 as a yellow powder
(30 mg, 90%). Mp 162 ꢀC; 1H NMR (300 MHz; THF-d8):
2.46 (s, 3H,
d
CH3), 6.80–6.83 (m, 2H, 3D, 5D), 6.91–6.94 (m, 2H, 2D, 6D), 7.06 (dd,
Jo ¼ 8.0 Hz, Jm ¼ 1.5 Hz, 1H, 4C), 7.25 (s, 1H, 5A), 7.40 (br t, Jo ¼ 8.0 Hz,
1H, 5C), 7.77–7.78 (m, 2H, 2C, 6C), 7.93–8.00 (m, 4H, 2B, 3B, 5B, 6B,),
9.65 (s,1H, NH); 13C NMR (75 MHz; THF-d8):
d 22.5 (CH3),107.6 (5A),
115.4 (2C), 116.2 (3D, 5D), 117.8 (4B), 118.7 (2B, 6B), 119.5 (4C), 120.7
(6C), 120.9 (2D, 6D), 127.3 (3B, 5B), 129.6 (5C), 138.7 (1C), 143.1 (1B),
148.6 (1D), 154.2 (4D), 156.8 (CN4H), 159.1 (3C), 159.7 (2A), 164.6 (4A),
167.6 (6A); HRMS (CI) m/z: calcd for C24H20N7O2 438.1678 [M þ H]þ;
found 438.1675.
1H NMR (300 MHz; MeOD):
13C NMR (75 MHz; MeOD):
d
7.06–7.09 (m, 2H), 7.55–7.58 (m, 2H);
105.1 (4B), 120.6 (CN), 125.5 (2B, 6B),
d
134.4 (3B, 5B), 154.9 (1B), 156.9 (NH–C(NH)–NH2); MS (CI) m/z: 161
[M þ H]þ; Anal. Calcd for C23H20O3: C, 59.99; H, 5.03; N, 34.98.
Found: C, 59.22; H, 5.06; N, 34.15
4.2.24. 4-(3-(2-(4-(2H-Tetrazol-5-yl)phenylamino)-6-
methylpyrimidin-4-yl)phenoxy)phenyl dibenzyl phosphate (38)
Dry tetrachloromethane (2 mL) was added to a solution of 37
(67 mg, 0.153 mmol) in dry acetonitrile (10 mL) cooled at ꢁ15 ꢀC
and the resulting mixture was stirred at ꢁ15 ꢀC for 5 min. Diiso-
4.2.21. 4-{4-[3-(4-Benzyloxyphenoxy)-phenyl]-6-methylpyrimidin-
2-ylamino}-benzo-1-nitrile (35)
A mixture of aryloxyphenyl butenone 31 (308 mg, 0.89 mmol)
and aryl guanidine 34 (430 mg, 2.68 mmol) in N,N-dimethylaceta-
mide (10 mL) was heated at 160 ꢀC for 6 h under air atmosphere. It
was cooled overnight to room temperature. Water was added and
the mixture was extracted with ether. The combined extracts were
washed with water and brine, dried over magnesium sulfate and
evaporated under reduced pressure to give an orange oil. Purifi-
cation by flash chromatography with cyclohexane/ethyl acetate
(5:2) gave 35 as a white powder (350 mg, 81%). Mp 138 ꢀC (recryst.
propylethylamine (200 mL, 1.15 mmol) and N,N-dimethylamino-
pyrimidine (10 mg, 0.082 mmol) were then added and the mixture
was stirred at ꢁ15 ꢀC for 30 min. Dibenzyl phosphite (180
mL,
0.815 mmol) was added dropwise over 45 min, the mixture was
stirred at ꢁ15 ꢀC for 1 h and then quenched with a 0.5 M solution
of potassium dihydrogen phosphate (4 mL). The mixture was
warmed up to room temperature and extracted with ethyl acetate.
The combined extracts were then washed with water and brine,
dried over magnesium sulfate and evaporated under reduced
pressure to give a yellow oil. Purification by flash chromatography
with dichloromethane/methanol (98:2) afforded 38 as a pale
yellow solid (65 mg, 61%). Mp 77–80 ꢀC; 1H NMR (300 MHz;
from ethyl acetate/pentane); 1H NMR (300 MHz; THF-d8):
d 2.43 (s,
3H, CH3), 5.07 (s, 2H, CH2), 7.01 (s, 4H 2D, 3D, 5D, 6D), 7.06 (ddd,
Jo ¼ 8.1 Hz, Jm ¼ 2.5 Hz, Jm ¼ 2.5 Hz, 1H, 4C), 7.23–2.35 (m, 4H, 5A, 3E,
4E, 5E), 7.40–7.44 (m, 3H, 5C, 2E, 6E), 7.54–7.58 (m, 2H, 3B, 5B), 7.77–
7.79 (m, 2H, 2C, 6C), 7.95–7.98 (m, 2H, 2B, 6B), 9.23 (s, 1H, NH); 13C
CDCl3):
d 2.47 (s, 3H, CH3), 5.16 (s, 2H, CH2), 5.19 (s, 2H, CH2) 7.06–
NMR (75 MHz; THF-d8):
d
23.3 (CH3), 70.2 (CH2), 103.7 (4B), 108.4
7.09 (m, 2H, 3D, 5D), 7.12 (s, 1H, 5A), 7.16–7.18 (m, 2H, 2D, 6D), 7.31
(m, 11H, 2 ꢂ 2E, 2 ꢂ 3E, 2 ꢂ 4E, 2 ꢂ 5E, 2 ꢂ 6E, 4C), 7.47–7.52 (m, 1H,
5C), 7.64–7.66 (m, 3H, 2B, 6B, 6C), 7.86–7.89 (m, 3H, 3B, 5B, 2C); 13C
(5A), 115.8 (3D, 5D), 115.9 (2C), 118.2 (2B, 6B), 118.8 (CN), 119.7 (4C),
120.5 (2D, 6D), 121.0 (6C), 127.2 (2E, 6E), 127.5 (4E), 128.1 (3E, 5E),
129.8 (5C), 132.6 (3B, 5B), 137.5 (1E), 139.1 (1C), 145.1 (1B), 150.2 (1D),
155.5 (4D), 159.1 (3C), 159.9 (4A), 163.8 (6A), 168.9 (2A); MS (ESþ) m/
z: 485 [M þ H]þ, 507 [M þ Na]þ. Anal. Calcd for C31H24N4O2: C,
76.84; H, 4.99; N, 11.56. Found: C, 76.62; H, 4.88; N, 11.39.
NMR (75 MHz; CDCl3):
d 24.0 (CH3), 70.7 (CH2), 70.8 (CH2), 109.1
(5A), 117.1 (2C), 117.4 (4B), 118.5 (2B, 6B), 119.3 (3D, 5D), 121.2 (2D, 6D),
122.3 (6C), 122.4 (4C), 128.1 (2E, 6E), 128.2 (3B, 5B), 128.7 (4E), 128.9
(3E, 5E), 130.4 (5C), 134.6 (1E), 139.1 (1C), 142.2 (1B), 145.5 (1D),

254
C. Courme et al. / European Journal of Medicinal Chemistry 45 (2010) 244–255
154.5, 154.7 (4D, CN4H), 157.2 (3C), 159.3 (2A), 164.2 (4A), 168.7 (6A);
Appendix. Supplementary data
31P NMR (121 MHz, CDCl3):
d
ꢁ5.60; MS (FABþ) m/z: 698 [M þ H]þ.
Supplementary data associated with this article can be found in
the online version, at doi:10.1016/j.ejmech.2009.10.003.
4.2.25. 4-(3-(2-(4-(2H-Tetrazol-5-yl)phenylamino)-6-
methylpyrimidin-4-yl)phenoxy)phenyl dihydrogen phosphate (4)
10% Palladium on carbon (3 mg) was added to a solution of 38
(30 mg, 0.043 mmol) in methanol (5 mL). The mixture was stirred
under H2 for 10 days at room temperature, then filtered through
celite and rinsed with methanol and water. The combined filtrates
were evaporated under reduced pressure. Purification by flash
chromatography with acetonitrile/water/acetic acid (100:8:2) gave
4 as a white solid (11 mg, 49%). Mp 168–170 ꢀC; 1H NMR (300 MHz;
References
[1] For a review of bioactive pyrimidines, see: D. Fabbro, S. Ruetz, E. Buchdunger,
S.W. Cowan-Jacob, G. Fendrich, J. Liebetanz, J. Mestan, T. O’Reilly, P. Traxler,
B. Chaudhuri, H. Fretz, J. Zimmerman, T. Meyer, G. Caravatti, P. Furet,
P.W. Manley Pharmacol. Ther. 93 (2002) 79–98.
[2] J. Zimmermann, E. Buchdunger, H. Mett, T. Meyer, N.B. Lydon, Bioorg. Med.
Chem. Lett. 7 (1997) 187–192.
[3] S.G. O’Brien, F. Guilhot, R.A. Larson, I. Gathmann, M. Baccarani, F. Cervantes,
J.J. Cornelissen, T. Fischer, A. Hochhaus, T. Hughes, K. Lechner, J.L. Nielsen,
P. Rousselot, J. Reiffers, G. Saglio, J. Shepherd, B. Simonsson, A. Gratwohl,
J.M. Goldman, H. Kantarjian, K. Taylor, G. Verhoef, A.E. Bolton, R. Capdeville,
B.J. Druker, N. Engl. J. Med. 348 (2003) 994–1004.
[4] S. Wang, G. Wood, C. Meades, G. Griffiths, C. Midgley, I. McNae, C. McInnes,
S. Anderson, W. Jackson, M. Mezna, R. Yuill, M. Walkinshaw, P. Fischer, Bioorg.
Med. Chem. Lett. 14 (2004) 4237–4240.
[5] X.-J. Chu, W. DePinto, D. Bartkovitz, S.-S. So, B.T. Vu, K. Packman, C. Lukacs,
Q. Ding, N. Jiang, K. Wang, P. Goelzer, X. Yin, M.A. Smith, B.X. Higgins, Y. Chen,
Q. Xiang, J. Moliterni, G. Kaplan, B. Graves, A. Lovey, N. Fotouhi, J. Med. Chem.
49 (2006) 6549–6560.
[6] U. Lu¨cking, G. Siemeister, M. Scha¨fer, H. Briem, M. Kru¨ger, P. Lienau, R. Jautelat,
Chem. Med. Chem. 2 (2007) 63–77.
DMSO-d6):
d 2.41 (s, 3H, CH3), 6.93–6.96 (m, 2H, 3D, 5D), 7.07–7.09
(m, 1H, 4C), 7.17–7.19 (m, 2H, 2D, 6D), 7.28 (s, 1H, 5A), 7.47–7.52 (m,
1H, 5C), 7.75–7.78 (m, 3H, 2B, 6B, 2C), 7.83–7.85 (m, 3H, 3B, 5B, 6C),
9.61 (s, 1H, NH); 13C NMR (75 MHz; DMSO-d6):
d 24.3 (CH3), 108.0
(5A), 116.0 (2C), 118.9 (2B, 6B), 120.1 (3D, 5D), 120.2 (4C), 121.5 (2D, 6D),
121.7 (6C), 125.4 (4B), 126.7 (3B, 5B), 130.9 (5C), 139.3 (1C), 140.4 (1B),
150.2 (4D), 151.5 (1D), 158.8 (3C), 160.4 (2A), 160.6 (CN4H), 163.2 (4A),
169.1 (6A); 31P NMR (121 MHz; DMSO-d6):
d
ꢁ5.06; MS (ESꢁ) m/z:
516 [M ꢁ H]ꢁ, 436 [M ꢁ PO3H2]ꢁ; HRMS (ESꢁ) m/z: calcd for
C24H20N7O5P 516.1185 [M ꢁ H]ꢁ; found 516.1185.
[7] M.W. Martin, J. Newcomb, J.J. Nunes, D.C. McGowan, D.M. Armistead,
C. Boucher, J.L. Buchanan, W. Buckner, L. Chai, D. Elbaum, L.F. Epstein, T. Faust,
S. Flynn, P. Gallant, A. Gore, Y. Gu, F. Hsieh, X. Huang, J.H. Lee, D. Metz,
S. Middleton, D. Mohn, K. Morgenstern, M.J. Morrison, P.M. Novak, A. Oliveira-
dos-Santos, D. Powers, P. Rose, S. Schneider, S. Sell, Y. Tudor, S.M. Turci,
A.A. Welcher, R.D. White, D. Zack, H. Zhao, L. Zhu, X. Zhu, C. Ghiron,
P. Amouzegh, M. Ermann, J. Jenkins, D. Johnston, S. Napier, E. Power, J. Med.
Chem. 49 (2006) 4981–4991.
[8] For a review on applications of cyanuric chloride in organic synthesis, see:
G. Blotny Tetrahedron 62 (2006) 9507–9522 and references 1–6 cited therein.
[9] H.-S. Moon, E.M. Jacobson, S.M. Khersonsky, M.R. Luzung, D.P. Walsh,
W. Xiong, J.W. Lee, P.B. Parikh, J.C. Lam, T.-W. Kang, G.R. Rosania, A.F. Schier, Y.-
T. Chang, J. Am. Chem. Soc. 124 (2002) 11608–11609.
[10] R. Menicagli, S. Samaritani, G. Signore, F. Vaglini, L. Dalla Via, J. Med. Chem. 47
(2004) 4649–4652.
[11] S. Mandal, G. Be´rube´, E. Asselin, I. Mohammad, V.J. Richardson, A. Gupta,
S.K. Pramanik, A.L. Williams, S.K. Mandal, Bioorg. Med. Chem. Lett. 17 (2007)
4955–4960.
[12] N. Baindur, N. Chadha, B.M. Brandt, D. Asgari, R.J. Patch, C. Schalk-HiHi,
T.E. Carver, I.P. Petrounia, C.A. Baumann, H. Ott, C. Manthey, B.A. Springer,
M.R. Player, J. Med. Chem. 48 (2005) 1717–1720.
[13] B.L. Hodous, S.D. Geuns-Meyer, P.E. Hughes, B.K. Albrecht, S. Bellon, J. Bready,
S. Caenepeel, V.J. Cee, S.C. Chaffee, A. Coxon, M. Emery, J. Fretland, P. Gallant,
Y. Gu, D. Hoffman, R.E. Johnson, R. Kendall, J.L. Kim, A.M. Long, M. Morrison,
P.R. Olivieri, V.F. Patel, A. Polverino, P. Rose, P. Tempest, L. Wang,
D.A. Whittington, H. Zhao, J. Med. Chem. 50 (2007) 611–626.
[14] A. Agarwal, K. Srivastava, S.K. Puri, P.M.S. Chauhan, Bioorg. Med. Chem. Lett. 15
(2005) 531–533.
4.2.26. Sodium 5-(4-(4-methyl-6-(3-(4-(phosphonatooxy)
phenoxy)phenyl)pyrimidin-2-ylamino)phenyl)-tetrazol-2-ide (39)
A 1 M sodium methoxide solution (80 mL, 0.080 mmol) was
added to a suspension of 4 (13 mg, 0.025 mmol) in dry methanol
(3 mL) at 0 ꢀC. The mixture was stirred at 0 ꢀC for 15 min and then
at room temperature for 1 h. The methanol was removed under
reduced pressure to afford 39 as a water-soluble white powder
(15 mg, quant.). 1H NMR (300 MHz; D2O):
d 2.24 (s, 3H, CH3), 6.88–
6.91 (m, 2H, 3D, 5D), 6.95 (s, 1H, 5A), 7.00 (d, Jo ¼ 8.1 Hz, 1H, 4C),
7.05–7.07 (m, 2H, 2D, 6D), 7.30–7.36 (m,1H, 5C), 7.41 (s,1H, 2C), 7.54–
7.60 (m, 3H, 2B, 6B, 6C), 7.76–7.79 (m, 2H, 3B, 5B); 13C NMR (75 MHz;
D2O):
d 21.2 (CH3), 108.6 (5A), 116.0 (2C), 118.9 (2B, 6B), 119.6 (4C),
120.4 (3D, 5D), 121.7 (2D, 6D), 122.0 (6C), 122.2 (4B), 126.8 (3B, 5B),
130.1 (5C), 137.8 (1C), 140.2 (1B), 150.2 (1D), 150.7 (4D), 157.9 (3C),
158.7 (2A), 161.6 (CN4H), 163.5 (4A), 169.2 (6A); 31P NMR (121 MHz;
D2O):
d
ꢁ0.60; MS (ESþ) m/z 584 [M þ H]þ, 562 [M ꢁ Na þ H]þ; MS
(ESꢁ) m/z: 582 [M ꢁ H]ꢁ, 538 [M ꢁ 2Na ꢁ H]ꢁ.
4.3. Competition assay
[15] A. Baliani, G.J. Bueno, M.L. Stewart, V. Yardley, R. Brun, M.P. Barrett, I.H. Gilbert,
J. Med. Chem. 48 (2005) 5570–5579.
[16] W. Huang, W. Zheng, D.J. Urban, J. Inglese, E. Sidransky, C.P. Austin, C.J. Thomas,
Bioorg. Med. Chem. Lett. 17 (2007) 5783–5789.
[17] For reviews, see: H. Fretz, P. Furet, C. Garcı´a Echeverrı´a, J. Rahuel, J. Schoepfer
Curr. Pharm. Des. 6 (2000) 1777–1796.
[18] C.B. Vu, Curr. Med. Chem. 7 (2000) 1081–1100.
[19] M. Vidal, V. Gigoux, C. Garbay, Crit. Rev. Oncol. Hematol. 40 (2001) 175–186.
[20] W.C. Shakespeare, Curr. Opin. Chem. Biol. 5 (2001) 409–415.
[21] T.R. Burke Jr., K. Lee, Acc. Chem. Res. 36 (2003) 426–433.
[22] K. Machida, B.J. Mayer, Biochim. Biophys. Acta 1747 (2005) 1–25.
[23] For the crystal structure of the mammalian Grb2 adaptor which is deposited
in the Protein Data Bank under the accession code 1GRI, see: S. Maignan,
J.P. Guilloteau, N. Fromage, B. Arnoux, J. Becquart, A. Ducruix Science 268
(1996) 291–293.
Precoated streptavidin plates (Boehringer) were incubated with
100 mL/well of biotin-Ahx-PSpYVNVQN peptide (100 nM in PBS
buffer) overnight at 4 ꢀC. Nonspecific binding was blocked with
PBS/3% BSA for 4 h at 4 ꢀC. Competitors were incubated at the
appropriate concentrations in PBS/3% milk containing 40 nM GST-
Grb2 protein (100 m
L/well) overnight at 4 ꢀC. Revelation is made
after anti-GST (Transduction Laboratories; 1/500 in PBS/milk/0.05%
Tween 20) and peroxidase-coupled antimouse (Amersham; 1/1000
in PBS/milk/0.05% Tween 20) incubations, using TMB solution
(Interchim). After coloration was stopped with H2SO4 (10% v/v), the
optical density (OD) was read at 550 nm. Dose–response relation-
ships were constructed by nonlinear regression of the competition
curves with Origin 40 software.
[24] C. Courme, S. Gillon, N. Gresh, M. Vidal, C. Garbay, J.-C. Florent,
E. Bertounesque, Tetrahedron Lett. 49 (2008) 4542–4545.
[25] For molecular modeling of ligand 1 binding mode to Grb2 SH2, see: W.-Q. Liu,
M. Vidal, N. Gresh, B.P. Roques, C. Garbay J. Med. Chem. 42 (1999) 3737–3741.
[26] For the X-ray crystallographic data of the ligand 1 binding to Grb2 SH2, see:
P. Nioche, W.-Q. Liu, I. Boutin, F. Charbonnier, M.-T. Latreille, M. Vidal,
B.P. Roques, C. Garbay, A. Ducruix J. Mol. Biol. 315 (2002) 1167–1177.
[27] W.-Q. Liu, M. Vidal, C. Olszowy, E. Million, C. Lenoir, H. Dhotel, C. Garbay, J.
Med. Chem. 47 (2004) 1223–1233.
[28] A.P. Benfield, M.G. Teresk, H.R. Plake, J.E. DeLorbe, L.E. Millspaugh, S.F. Martin,
Angew. Chem. 118 (2006) 6984–6989.
[29] A.P. Benfield, M.G. Teresk, H.R. Plake, J.E. Delorbe, L.E. Millspaugh, S.F. Martin,
Angew. Chem. Int. Ed. Engl. 45 (2006) 6830–6835.
Acknowledgements
`
We thank CNRS, Institut Curie and Ministere de la Recherche
ꢀ
´
´
(ACI ‘‘Molecules et Cibles Therapeutiques’’ 2002 N 02L0521) for
´
financial support. Fondation pour la Recherche Medicale (FRM) is
gratefully acknowledged for a fellowship granted to Caroline
Courme.

C. Courme et al. / European Journal of Medicinal Chemistry 45 (2010) 244–255
255
[30] X ray crystallographic data are deposited in the Protein Data Bank under the
accession codes 2HUW and 2HUY. This latter was obseleted on 2008-08-12
and superceded by 3C7I.
[31] For reviews on tetrazoles as carboxylic acid isosteres, see: G.A. Patani,
E.J. LaVoie Chem. Rev. 96 (1996) 3147–3176.
[66] R.A. Daines, K.K.C. Sham, J.J. Taggart, W.D. Kingsbury, J. Chan, A. Breen, J. Disa,
N. Aiyar, Bioorg. Med. Chem. Lett. 20 (1997) 2673–2676.
[67] M. Koelbel, T. Beyersdorff, C. Tschierske, S. Diele, J. Kain, Chem. Eur. J. 20
(2000) 3821–3837.
[68] T.Y.H. Wu, S. Ding, N.S. Gray, P.G. Schultz, Org. Lett. 3 (2001) 3827–3830.
[69] M.B. Steffensen, E.E. Simanek, Org. Lett. 5 (2003) 2359–2361.
[70] For a previous work on the displacement of aryl fluorides with hydroquinone,
see: B.F. Marcune, M.C. Hillier, J.-F. Marcoux, G.R. Humphrey Tetrahedron Lett.
46 (2005) 7823–7826.
[32] R.J. Herr, Bioorg. Med. Chem. 10 (2002) 3379–3393.
[33] A Scifinder search on the triazine scaffold incorporating a bis-aryl phosphate
exclusively identified two patents, on triazine dyes, see: B.L. McConnell, R.
Thornton, L.A. Graham, U.S. US 4091004 19780523; 1978 and ref. [34].
[34] K. Sato, H. Tanaka, Jpn. Kokai Tokkyo Koho. JP 04117369 A 19920417 Heisei.
CAN 117:152766 AN 1992:552766; 1992.
[71] S.H. Szajnman, W. Yan, B.N. Bailey, R. Docampo, E. Elhalem, J.B. Rodriguez, J.
Med. Chem. 43 (2000) 1826–1840.
[35] For example, see: R.T. Timmer, C.W. Alexander, S. Pillarisetti, U. Saxena, K.R.
Yeleswarapu, M. Pal, J.T. Reddy, V.V.R.M.K. Reddy, B.S. Sridevi, P.R. Kumar, G.O.
Reddy, PCT Int. Appl. WO 2004026844; 2004.
[36] For compound 8, see: A Scifinder search exclusively identified three patents. P.
Furet, P. Imbach, T.M. Ramsey, A. Schlapbach, D. Scholz, G. Caravatti, PCT Int.
Appl. WO 2004005282; 2004.
[37] For compound 9, see: S. Jones, B. Westwood, M. Thomas, J. McLachlan, K.
Duncan, F. Scaerou, D. Zheleva, PCT Int. Appl. WO 2007042784; 2007.
[38] For compound 10, see: C.F. Claiborne, L.J. Payne, R.J. Boyce, T.B. Sells, S.G.
Stroud, S. Travers, T.J. Vos, G.S. Weatherhead, U.S. Pat. Appl. Publ. US
20050256102; 2005.
[72] The trimer derivative [70] – 1,4-bis(3-bromophenoxy)benzene – was not
formed under these basic conditions. However, the desired product 12 was
isolated along with an inseparable mixture of a bis(bromophenyl)-[1,4]ben-
zoquinone [MS (CI, DCI/NH3): m/z 417, 419, 421], resulting from the reaction of
hydroquinone as an aryloxide C-nucleophile, and an unidentified compound.
[73] For an alternative method (i.e., a copper-catalyzed arylboronic acid-hetero-
atom coupling) to access such a boronic acid, see: T.E. Pennington, C. Kardi-
man, C.A. Hutton Tetrahedron Lett. 45 (2004) 6657–6660.
[74] L.J. Silverberg, J.L. Dillon, P. Vemishetti, Tetrahedron Lett. 37 (1996) 771–774.
[75] For an adaptation of the phosphorylation procedure described by Silverberg
et al. [74] for solid-phase synthesis, see: P. Deprez, E. Mandine, D. Gofflo,
S. Meunier, D. Lesuisse Bioorg. Med. Chem. Lett. 12 (2002) 1295–1298.
[76] P.K. Grzyska, P.G. Czyryca, J. Purcell, A.C. Hengge, J. Am. Chem. Soc. 2125 (2003)
13106–13111 and references cited therein.
[77] A.J. Cocuzza, F.W. Hobbs, C.R. Arnold, D.R. Chidester, J.A. Yarem, S. Culp,
L. Fitzgerald, P.J. Gilligan, Bioorg. Med. Chem. Lett. 9 (1999) 1057–1062.
[78] L. Strekowski, D.B. Harden, W.B. Grubb III, S.E. Patterson, A. Czarny,
M. Mokrosz, M.T. Cegla, R. Wydra, J. Heterocycl. Chem. 27 (1990) 1393–1400.
[79] M.G. Bursavich, S. Lombardi, A.M. Gilbert, Org. Lett. 7 (2005) 4113–4116.
[80] A.J. Cocuzza, D.R. Chidester, S. Culp, L. Fitzgerald, P.J. Gilligan, Bioorg. Med.
Chem. Lett. 9 (1999) 1063–1066.
[39] O. Diels, Ber. Dtsch. Chem. Ges. 32 (1899) 691–702.
[40] H. Koopman, J. Daams, Recl. Trav. Chim. Pays-Bas 77 (1958) 235–240.
[41] M. Goi, Yuki Gosei Kagaku Kyokaishi 18 (1960) 327–331.
[42] M. Goi, Yuki Gosei Kagaku Kyokaishi 18 (1960) 332–336.
[43] A.R. Katritzky, D.C. Oniciu, I. Ghiviriga, R.A. Barcock, J. Chem. Soc. Perkin Trans.
2 (1995) 785–792.
[44] P. de Hoog, P. Gamez, W.L. Driessen, J. Reedijk, Tetrahedron Lett. 43 (2002)
6783–6786.
[45] K. Leftheris, G. Ahmed, R. Chan, A.J. Dyckman, Z. Hussain, K. Ho, J. Hynes Jr.,
J. Letourneau, W. Li, S. Lin, A. Metzger, K.J. Moriarty, C. Riviello, Y. Shimshock,
J. Wen, J. Wityak, S.T. Wrobleski, H. Wu, J. Wu, M. Desai, K.M. Gillooly, T.H. Lin,
D. Loo, K.W. McIntyre, S. Pitt, D.R. Shen, D.J. Shuster, R. Zhang, D. Diller,
A. Doweyko, J. Sack, J. Baldwin, J. Barrish, J. Dodd, I. Henderson, S. Kanner,
G.L. Schieven, M. Webb, J. Med. Chem. 47 (2005) 6283–6291.
[46] A. Baliani, G.J. Bueno, M.L. Stewart, V. Yardley, R. Brun, M.P. Barrett, I.H. Gilbert,
J. Med. Chem. 48 (2005) 5570–5579.
[81] W. Wendelin, K. Schermanz, J. Heterocycl. Chem. 21 (1984) 65–69 and refer-
ences cited therein.
[82] T.J. Delia, J.M. Schomaker, A.S. Kalinda, J. Heterocycl. Chem. 43 (2006) 127–131.
[83] For an example of the Heck reaction of halogeno-diphenyl ethers with ethyl
acrylate, see: G.F. Orr, D.L. Musso, J.L. Kelley, S.S. Joyner, S.T. Davis,
D.P. Baccanari J. Med. Chem. 40 (1997) 1179–1185.
[47] S.R. Sandler, J. Org. Chem. 35 (1970) 3967–3968 and references cited therein.
[48] R. Menicagli, C. Malanga, P. Peluso, Synth. Commun. 24 (1994) 2153–2158.
[49] H. Koopman, J.H. Uhlenbroek, H.H. Haeck, J. Daams, M.J. Koopmans, Recl. Trav.
Chim. Pays-Bas 78 (1959) 967–978.
[84] X. Xu, G. Fakha, D. Sinou, Tetrahedron 58 (2002) 7539–7544.
[85] For the thermal polymerization of a vinyl ketone during the Heck reaction,
see: H. Uchiro, K. Nagasawa, Y. Aiba, S. Kobayashi Tetrahedron Lett. 41 (2000)
4165–4168.
[50] J.T. Bork, J.W. Lee, S.M. Khersonsky, H.-S. Moon, Y.-T. Chang, Org. Lett. 5 (2003)
117–120.
[51] For the preparation of 2,4,6-tricyano-1,3,5-triazine (TCT), see: G. Beck, United
States Patent 5086172; 1992.
[86] For the value of
a,b-unsaturated ketones as key intermediates for the
synthesis of pyrimidines on the solid phase, see: A.L. Marzinzik, E.R.J. Felder
Org. Chem. 63 (1998) 723–727.
[87] V.J. Bauer, S.R. Safir, J. Med. Chem. 9 (1966) 244–246.
[88] D. Amantini, R. Beleggia, F. Fringuelli, F. Pizzo, L. Vaccaro, J. Org. Chem. 69
(2004) 2896–2898.
[89] W.-Q. Liu, M. Vidal, C. Olszowy, E. Million, C. Lenoir, H. Dhotel, C. Garbay, J.
Med. Chem. 47 (2004) 1223–1233.
[90] Very recently, Lo
´ pez-Rodrı´guez et al. disclosed a non-peptide inhibitor of
[52] H. Yamanaka, S. Ohba, S. Konno, Heterocycles 26 (1987) 2853–2856.
[53] R. Faust, B. Gobelt, Tetrahedron Lett. 38 (1997) 8017–8020.
[54] S.Samaritani,G.Signore,C.Malanga,R.Menicagli,Tetrahedron61(2005)4475–4483.
[55] To the best of our knowledge, only one report mentioned the mono-
coupling reaction of organoboronic acids with 2,4,6-trichloro-1,3,5-triazine,
see: J.-Q. Tan, J.-H. Chang, M.-Z. Deng Chin. J. Chem. 22 (2004) 941–944.
[56] D. Janietz, M. Bauer, Synthesis (1993) 33–34.
[57] J.T. Bork, J.W. Lee, Y.-T. Chang, Tetrahedron Lett. 44 (2003) 6141–6144.
[58] G. Cooke, H. Augier de Cremiers, V.M. Rotello, B. Tarbit, P.E. Vanderstraeten,
Tetrahedron 57 (2001) 2787–2789.
[59] C.A.G.N. Montalbetti, T.S. Coulter, M.K. Uddin, S.G. Reignier, F. Magaraci,
C. Grana¨s, C. Krog-Jensen, J. Felding, Tetrahedron Lett. 47 (2006) 5973–5975.
[60] G.-H. Kuo, A. DeAngelis, E. Stuart, A. Wang, Y. Zhang, P.J. Connolly, X. Chen,
R.H. Gruninger, C. Rugg, A. Fuentes-Pesquera, S.A. Middleton, L. Jolliffe,
W.V. Murray, J. Med. Chem. 48 (2005) 4535–4546.
[61] R. Menicagli, S. Samaritani, V. Zucchelli, Tetrahedron 56 (2000) 9705–9711 and
references cited therein.
[62] G.-S. Yang, X.-J. Xie, G. Zhao, Y. Ding, J. Fluorine Chem. 98 (1999) 159–162.
[63] R.P. Robinson, E.R.J. Laird, F. Blake, J. Bordner, K.M. Donahue, L.L. Lopresti-
Morrow, P.G. Mitchell, M.R. Reese, L.M. Reeves, E.J. Stam, S.A. Yocum, J. Med.
Chem. 43 (2000) 2293–2296.
[64] R.K. Juneja, K.D. Robinson, C.P. Johnson, J.L. Atwood, J. Am. Chem. Soc. 115
(1993) 3818–3819.
[65] P.J. Hajduk, J. Dinges, G.F. Miknis, M. Merlock, T. Middleton, D.J. Kempf,
D.A. Egan, K.A. Walter, T.S. Robins, S.B. Shuker, T.F. Holzman, S.W. Fesik, J. Med.
Chem. 40 (1997) 3144–3150.
Grb2-SH2, namely UCM104, as a new lead structure, which has also an IC50
M): see, XXth International
value in the micromolar range (IC50 ¼ 174
m
Symposium on Medicinal Chemistry, August 31- September 4, 2008, Vienna,
Austria. Poster Presentation P527 Development of non-peptide inhibitors of
Her2-Grb2 interaction using molecular modeling and NMR spectroscopy. M.L.
´
´
´
´
Lopez-Rodriguez, S. Ortega-Gutierrez, B. Benhamu, A.L. Orcajo, P. Serrano,
A.Viso, I.R. Torrecillas, M. Campillo, L. Pardo, K. Wu¨thrich.
[91] See Supporting information.
[92] Low-temperature molecular dynamics (300 steps of
5 picoseconds each,
T ¼ 100 K, ¼ 4.) were done with the Discover module and the Cff91 force-
3
field of the Insight II package (Accelrys, Inc). We resorted to the crystal
structure of the complex of 1 with Grb2-SH2 to dock 1 and all non-peptidic
compounds in the binding site.
[93] Y.T. Chen, C.T. Seto, Bioorg. Med. Chem. 12 (2004) 3289–3298.
[94] J. Xie, C.T. Seto, Bioorg. Med. Chem. 15 (2007) 458–473.
[95] A.P. Combs, E.W. Yue, M. Bower, P.J. Ala, B. Wayland, B. Douty, A. Takvorian,
P. Polam, Z. Wasserman, W. Zhu, M.L. Crawley, J. Pruitt, R. Sparks, B. Glass,
D. Modi, E. McLaughlin, L. Bostrom, M. Li, L. Galya, K. Blom, M. Hillman,
L. Gonneville, B.R. Reid, M. Wei, M. Becker-Pasha, R. Klabe, R. Huber, Y. Li,
G. Hollis, T.C. Burn, R. Wynn, P. Liu, B. Metcalf, J. Med. Chem. 48 (2005) 6544–
6548.