PYRAZOLE DERIVATIVES, PREPARATION THEREOF AND THERAPEUTIC
USE THEREOF
[0001] The present invention relates to pyrazole derivatives, to the preparation
thereof and to the therapeutic use thereof.
[0002] More particularly, and according to a first aspect, the invention relates
to novel specific substituted pyrazoles with anticancer activity, via modulation of the
activity of proteins, in particular kinases.
[0003] Protein kinases are a family of enzymes that catalyse the
phosphorylation of hydroxyl groups of specific residues of proteins such as tyrosine,
serine or threonine residues. Such phosphorylations can largely modify the function of
proteins; thus, protein kinases play an important role in regulating a wide variety of cell
processes, especially including metabolism, cell proliferation, cell differentiation, cell
migration or cell survival. Among the various cellular functions in which the activity of a
protein kinase is involved, certain processes represent attractive targets for treating
cancer diseases and also other diseases.
[0004] Thus, one of the objects of the present invention is to propose
compositions with anticancer activity, by acting in particular with respect to kinases.
Among the kinases for which a modulation of activity is desired, mention may be
made of KDR, Tie2, VEGFR-1 (FLT1), VEGFR-3 (FLT4), PDGFR and FGFR. The
kinases KDR and/or Tie2 are preferred.
[0005] Compounds corresponding to the general formula (I) below are known
from the patent application published under the number WO 08/065282:
in which:
1) A and Ar are independently selected from the group constituted by: aryl,

heteroaryl, substituted aryl, substituted heteroaryl;
2) L is selected from the group constituted by: NH-CO-NH and O-CO-NH;
3) R1 is selected from the group constituted by: H, R6, COR6, SO2R6, in which R6 is
chosen from H, OR7, NR8R9, alkyl, cycloalkyl, heterocyclyl, substituted heterocyclyl,
aryl, substituted aryl, heteroaryl and substituted heteroaryl, in which R7 is chosen from
H, phenyl and alkyl, and in which R8 and R9 are independently selected from the group
constituted by H, alkyl, cycloalkyl, heterocyclyl, substituted heterocyclyl, aryl,
substituted aryl, heteroaryl and substituted heteroaryl, or alternatively R8 and R9 are
linked together to form a saturated 5- to 8-membered ring containing from 0 to 3
heteroatoms chosen from O, S and N;
4) X is selected from the group constituted by: O and NH;
5) R3 is selected from the group constituted by: H, alkyl, substituted alkyl, cycloalkyl,
substituted cycloalkyl;
6) R4a is selected from the group constituted by: H and (C1-C4)alkyl;
7) R4b is selected from the group constituted by: H and (C1-C4)alkyl;
8) R5 is selected from the group constituted by: H, halogen, R10, CN, O(R10), OC(O)
(R10), OC(O)N(R10)(R11), OS(O2)(R10), N(R10)(R11), N=C(R10)(R11), N(R10)C(O)(R11),
N(R10)C(O)O(R11), N(R12)C(O)N(R10)(R11), N(R12)C(S)N(R10)(R11), N(R10)S(O2)(R11), C(O)
(R10), C(O)O(R10), C(O)N(R10)(R11), C(=N(R11))(R10), C(=N(OR11))(R10), S(R10), S(O)(R10),
S(O2)(R10), S(O2)O(R10), S(O2)N(R10)(R11); in which each R10l R11, R12 is independently
selected from the group constituted by H, alkyl, alkylene, alkynyl, aryl, heteroaryl,
cycloalkyl, heterocyclyl, substituted alkyl, substituted alkylene, substituted alkynyl,
substituted aryl, substituted heteroaryl, substituted cycloalkyl, substituted heterocyclyl.
[0006] In this patent, preferably X is O, R3 is methyl, R4a and R4b are H; L is
NHCONH; A is phenyl; Ar is phenyl; but in this application no example describes the
substitution of Ar and its effect on the pharmacokinetics.
[0007] One subject of the present invention are two compounds included in the
preceding application that correspond to the formula (I):


in which: j
X represents CI or F.
[0008] The compounds of formula (I) may exist in the form of bases or acid-addition
salts. Such addition salts are part of the invention.
[0009] The compounds in their two tautomeric forms indicated below belong to the
invention:

[0010] These salts may be prepared with pharmaceutically acceptable acids, but the
salts of other acids that are of use, for example, for the purification or isolation of the
compounds of formula (I) are also part of the invention. Among the salts that can be
used, mention may especially be made of the hydrochloride.
[0011] The compounds of formula (I) may also exist in the form of hydrates or
solvates, namely in the form of associations or combinations with one or more water
molecules or with a solvent. Such hydrates and solvates are also part of the invention.
[0012] Among the compounds of formula (I) that are subjects of the invention,
mention may especially be made of the following compounds:
4-{3-[3-(2-chloro-4-trifluoromethylphenyl)ureido]-5-fluorobenzylamino}-1H-
pyrazole-3-carboxamide and its hydrochloride;
4-{3-[3-(2-fluoro-4-trifluoromethylphenyl)ureido]-5-fluorobenzylamino}-1H-

pyrrazole-3-carboxamide and its hydrochloride.
[0013] In accordance with the invention, it is possible to prepare the compounds of
general formula (I) according to the process that follows.
[0013] In the schemes which follow, the initial compounds and the reactants, when
their method of preparation is not described, are commercially available or are ;
described in the literature, or else may be prepared according to methods which are
described therein or which are known to a person skilled in the art.
[0014] According to another of its aspects, another subject of the invention are the!
compounds of formulae:

in which X represents F or CI.
These compounds are of use as synthetic intermediates of the compounds of formula
(I).
[0015] The following examples describe the preparation of the compounds ini
accordance with the invention. These examples are not limiting and serve only to
illustrate the present invention.
Process for synthesis of the examples


4-{3-[3-(2-Chloro-4-trifluoromethylphenyl)ureido]-5-fluorobenzylamino}-
1 H-pyrazole-3-carboxamide hydrochloride

A suspension of 1.64 g (3.48 mmol) of 4-{3-[3-(2-chloro-4-
trifluoromethylphenyl)ureido]-5-fluorobenzylamino}-1H-pyrazole-3-carboxamide in
50 ml of ethanol is stirred at ambient temperature under an argon atmosphere. Then

35 ml (35 mmol) of a solution of hydrochloric acid in diethyl ether (1 N) are added
dropwise. The reaction medium becomes a clear solution. After stirring for 10 hours at
ambient temperature, the solvents are evaporated using a rotary evaporator under
reduced pressure. The residue obtained is stirred in 50 ml of diethyl ether for
30 minutes.
After filtration and drying in an oven, 1.8 g of 4-{3-[3-(2-chloro-4-
trifluoromethylphenyl)ureido]-5-fluorobenzylamino}-1H-pyrazole-3-carboxamide
hydrochloride are obtained in the form of pale yellow crystals.
MS: Retention time Tr (min) = 1.01; [M+H]+ m/z = 471; [M-H]- m/z = 469
1H NMR (400 MHz, DMSO-d) δ ppm 4.32 (s, 2 H) 6.89 (broad d, J=9.6 Hz, 1 H) 7.16
(broad s, 1 H) 7.18 - 7.61 (m, 4 H) 7.68 (broad d, J=8.9 Hz, 1 H) 7.86 (broad s, 1 H) 8.44 (d,|
J=8.9 Hz, 1 H) 8.81 (broad m, 1 H) 10.17 (broad m, 1 H)
Melting point (Kofler) = 177°C
4-{3-[3-{2-Chioro-4-trifluoromethylphenyl)ureido]-5-fluorobenzylamino}-
1 H-pyrazole-3-carboxamide

A solution of 5.9 g (9.5 mmol) of 4-{3-[3-(2-chloro-4i
trifluoromethylphenyl)ureido]-5-fluorobenzylamino}-1H-pyrazole-3-(2,4-
dimethoxybenzylamide) and 4.5 g (24 mmol) of para-toluenesulphonic acid in 400 ml
of toluene is heated at reflux for 2 hours. After settling, the toluene solution is
separated from a yellow gum. The gum is diluted in 150 ml of methanol and 500 ml off
ethyl acetate. Then 500 ml of water are added. Next the solution is cooled, then
basified with 100 ml of an aqueous solution of potassium hydroxide (10 N). Aftelr
settling, the aqueous phase is extracted with a solution of 400 ml of ethyl acetate and
100 ml of methanol. The organic phases are recombined and washed with 100 ml of
a saturated sodium chloride solution, dried over magnesium sulphate and
concentrated under reduced pressure to give a pale yellow solid which is purified over

200 g of silica, eluted with an 80/10/10 (by volume) solution of
dichloromethane/methanol/acetonitrile: 1.77 g of 4-{3-[3-(2-chloro-4-
trifluoromethylphenyl)ureido]-5-fluorobenzylamino}-1H-pyrazole-3-carboxamide are
obtained in the form of a white solid.
MS: Retention time Tr (min) = 4.29; [M+H]+ m/z = 471; [M-H]m/z = 469
1H NMR (400 MHz, DMSO-d) δ ppm 4.19 (d, J=6.7 Hz, 2 H) δ.74 (broad m, 1 H) 6.80
(broad d, J=9.6Hz, 1 H) 6.95 - 7.11 (m, 3 H) 7.24 (broad unresolved m, 1 H) 7.42 (dt, J=11.3,
2.3 Hz, 1 H) 7.67 (broad d, /=8.9 Hz, 1H) 7.86 (broad s, 1 H) 8.44 (d, J=8.9 Hz, 1 H) 8.70
(broad s 1 H) 9.92 (broad s, 1 H) 12.57 (broad s, 1 H)
Melting point (Kofler): 220°C
4-{3-[3-(2-Chloro-4-trifluoromethylphenyl)ureido]-5-fluorobenzylamino}-
1 H-pyrazole-3-(2,4-dimethoxybenzylamide)

A solution of 10 g (32 mmol) of 4-amino-1H-pyrazole-3-(2,4-
dimethoxybenzylamide) hydrochloride and 5.9 ml (35.2 mmol) of
diisopropylethylamine in 300 ml of tetrahydrofuran is stirred at ambient temperature
under an argon atmosphere. 3.85 g (35 mmol) of magnesium sulphate and 12.70 g
(35.2 mmol) of 1-(2-chloro-4-trifluoromethylphenyl)-3-(3-fluoro-5-formylphenyl)urea
are added. The reaction mixture is then heated at reflux for 14 hours. The mixture is
then cooled to 25 °C, then 10.08 g (160 mmol) of sodium cyanoborohydride are slowly
added. After stirring for 12 hours at ambient temperature, the mixture is concentrated
to dryness using a rotary evaporator. The gum obtained is stirred with 300 ml of water
and 500 ml of an aqueous solution of sodium hydroxide (1N). This suspension is
stirred for 30 minutes with 1 L of dichloromethane. After filtration through No. 3

sintered glass, the solid obtained is rinsed with 2 x 500 ml of water then dried in an
oven under vacuum at 40°C. The solid is then recrystallized in 800 ml of methanol, at
high temperature, to give 8.3 g of 4-{3-[3-(2-chloro-4-trifluoromethylphenyl)ureido]-5-
f!uorobenzylamino}-1 H-pyrazole-3-(2,4-dimethoxybenzylamide) in the form of a white
powder.
MS: Retention time Tr (min) = 4.97; [M+H]+m/z = 621; [M-H]m/z = 619
'H NMR (400 MHz, DMSO-d δ ppm 3.73 (s, 3 H) 3.81 (s, 3 H) 4.19 (d, J=6.1 Hz, 21
H) 4.33 (d, J=6.7Hz, 2 H) δ.71 (broad t, ,J=6.7 Hz, 1 H) 6.46 (dd, J=8.3, 2.3 Hz, 1 H) 6.55 (d,j
J=2.3 Hz, 1 H) 6.80 (broad d,J=9.6 Hz, 1 H) 7.04 (broad s, 1 H) 7.08 (s, 1 H) 7.10 (d, J=8.9
Hz, 1 H) 7.43 (dt, J=11.4, 2.3 Hz, 1 H) 7.67(broad d, J=8.9 Hz, 1 H) 7.86 (broad s, 1 H) 7.94
(broad t, J=6.7 Hz, 1 H) 8.45 (d, J=8.9 Hz, 1 H) 8.62 (broad unresolved m, 1 H) 9.79 (broad
unresolved m, 1 H) 12.60 (broad unresolved m, 1 H)
Melting point (Kofler):168°C
1-(2-Chloro-4-trifluoromethylphenyl)-3-(3-fluoro-5-formylphenyl)urea

A solution of 33.15 g (92.68 mmol) of 1-(2-chloro-4-trifluoromethylphenyl)-3-(3-
cyano-5-fluorophenyl)urea in 600 ml of tetrahydrofuran is stirred at -10°C under ani
argon atmosphere. Next, 230 ml of a solution of diisobutylaluminium hydride at a
concentration of 20% in toluene are added with a regular "dropwise" action. The
reaction mixture is stirred at ambient temperature for 12 hours. Next, an additional
80 ml of diisobutylaluminium hydride are added at -10°C. After stirring for 14 hours at
ambient temperature, the reaction medium is concentrated to dryness using a rotary
evaporator in order to give a thick oil to which 500 g of ice and 100 ml of 100% acetic
acid are slowly added, while stirring. The suspension obtained is filtered. The solid is

then diluted in 2 x 800 ml of ethyl acetate and the organic solution is washed with
600 ml of a saturated sodium chloride solution, dried over magnesium sulphate,
filtered, concentrated to dryness using a rotary evaporator and dried in an oven under
vacuum at 40 °C in order to give 29.57 g of 1-(2-chloro-4-trifluoromethylphenyl)-3-(3-
fluoro-5-formylphenyl)urea in the form of a pale yellow powder.
MS: Retention time Tr (min) = 4.86; [M+H]+m/z = 361; [M-H]-m/z = 359
1H NMR (400 MHz, DMSO-d) δ ppra 7.35 (ddd, J=8.4, 2.3, 1.8 Hz, 1 H) 7.67 - 7.75
(m, 2 H) 7.78 (t, J=1.8 Hz, 1 H) 7.88 (d, J=1.8 Hz, 1 H) 8.45 (d, J=8.8 Hz, 1 H) 8.72 (broad s,
1 H) 9.98 (d, J=1.8 Hz, 1 H) 10.07 (broad s, 1 H)
1-(2-Chloro-4-trifluoromethylphenyl)-3-(3-cyano-5-fluorophenyl)urea

A solution of 15 g (110.2mmol) of 5-fluoro-3-cyanoaniline in 150 ml of:
tetrahydrofuran is stirred at ambient temperature under an argon atmosphere. Next,
17.5 ml (121.22mmol) of 2-chloro-4-trifluoromethylphenylisocyanate are added. The!
reaction medium is heated at reflux for 3 hours then concentrated to dryness using a
rotary evaporator. The solid residue obtained is recrystallized at high temperature in
60 ml of ethyl acetate in order to give 33.25 g of 1-(2-chloro-4-trifluoromethylphenyl)-j
3-(3-cyano-5-fluorophenyl)urea in the form of a white solid.
MS: Retention time Tr (min) = 4.97; [M+H]+: m/z = 356
Melting point (Kofler): 286°C
5-Fluoro-3-cyanoaniline is a commercial product.
4-Amino-1 H-pyrazole-3-(2,4-dimethoxybenzylamide) hydrochloride


A suspension of 6.12 g (20 mmol) of 4-nitro-1H-pyrazole-3-(2,4-
dimethoxybenzylamide) in 340 ml of ethanol is stirred at ambient temperature. Next,
15.8 g (70 mmol) of tin chloride dihydrate are added over 5 minutes. The reaction
mixture is stirred for 14 hours at ambient temperature, then concentrated to dryness
using a rotary evaporator. The residue obtained is stirred with 330 ml of an aqueous
solution saturated with sodiumhydrogen carbonate and 300 ml of dichloromethane.
After settling, the organic phase is extracted with 2 x 150 ml of dichloromethane. The!
organic phases are recombined, washed with 150 ml of a saturated sodium chloride!
solution and dried over magnesium sulphate. After concentrating using a rotary;
evaporator, 4.57 g of 4-amino-1H-pyrazole-3-(2,4-dimethoxybenzylamide)i
hydrochloride are obtained in the form of a solid of purplish colour.
MS: EI: [M]*. m/z = 276; base peak m/z = 151
'H NMR (400 MHz, DMSO-d) 8 ppm 3.73 (s, 3 H) 3.81 (s, 3 H) 4.31 (d, J=6.2 Hz, 1
H) 4.56 (broad unresolved m, 2 H) 6.46 (dd, J=8.3, 2.9 Hz, 1 H) 6.55 (d, J=2.9 Hz, 1 H) 7.06
7.13 (m, 2 H) 7.84 (t, J=6.2 Hz, 1 H) 12.50 (broad unresolved m, 1 H)
Melting point (Buchi): 186°C
4-Nitro-1H-pyrazole-3-(2,4-dimethoxybenzylamide)

A solution of 14.65 g (76.4 mmol) of 1 -(3-dimethylaminopropyl)-3-
ethylcarbodiimide dihydrate and 10.32g (76.4 mmol) of 1-hydroxybenzotriazole in
50 ml of dimethylformamide is stirred at ambient temperature. 11.7 g (70.03 mmol) of
2,4-dimethoxybenzylamine are added, then 10.2 g of 4-nitro-3-pyrazole carboxylic acid
are added in small portions. After 16 hours of stirring at ambient temperature, the

reaction medium is poured into 500 ml of water. The suspension is filtered, then
washed with 2 x 250 ml of water. The solid obtained is dried in an oven under vacuum
at 40°C in order to give 18.58 g of 4-nitro-1H-pyrazole-3-(2,4-dimethoxybenzylamide)
in the form of a white solid.
'H NMR (400 MHz, BMSO-d) δ ppm 3.75 (s, 3 H) 3.80 (s, 3 H) 4.35 (d, J=5.9 Hz, 2
H) 6.50 (dd, J=8.3, 2.4 Hz, 1 H) 6.56 (d, J=2.4 Hz, 1 H) 7.21 (d, J=8.3 Hz, 1 H) 8.71 (broad s,
1 H) 8.88 (broad t, J=5.9 Hz, 1 H) 14.13 (broad unresolved m, 1 H)
MS (ES+/-) Retention time Tr (min) = 3.23; [M+H]+ m/z = 307; [M-H]- m/z = 305
Melting point (Kofler):192°C
4-nitro-3-pyrazolecarboxylic acid is a commercial product.
Example 2
4-{3-[3-(2-Fluoro-4-trifluoromethylphenyl)ureido]-5-fluorobenzylamino}-
1 H-pyrazole-3-carboxamide hydrochloride

A suspension of 1.85 g (0.40 mmol) of 4-{3-[3-(2-fluoro-4t
trifluoromethylphenyl)ureido]-5-fluorobenzylamino}-1H-pyrazole-3-carboxamide irji
i
40 ml of ethanol is stirred at ambient temperature under an argon atmosphere. Theiji
20 ml (40 mmol) of a solution of hydrochloric acid in diethyl ether (1 N) are added
dropwise. The reaction medium becomes a clear solution. After stirring for 12 hours at
ambient temperature, the solvents are evaporated using a rotary evaporator under
reduced pressure. The residue obtained is stirred in 200 ml of diethyl ether for
30 minutes.
After filtration and drying in an oven, 1.75 g of 4-{3-[3-(2-fiuoro-4-
trifiuoromethylphenyl)ureido]-5-fluorobenzylamino}-1H-pyrazole-3-carboxamide

hydrochloride are obtained in the form of pale yellow crystals.
'H NMR (400 MHz, DMSO-tfc) 8 ppm 4.24 (s, 2 H) 6.83 (d, J=9.0 Hz, 1 H) 7.08 (s
1H) 7.18 (br. s., 1H) 7.23 (s, 1 H) 7.35 (br. s., 2 H) 7.42 (dt, J=11.3, 2.0 Hz, 1 H) 7.54 (d,J=8.8
Hz, 1 H) 7.69 (dd, J=11.2, 1.5 Hz, 1 H) 8.41 (t, J=8.2 Hz, 1 H) 8.99 (s, 1 H) 9.54 (s, 1 H)
MS: Retention time Tr (min) = 0.97; [M+H]+m/z = 455
Melting point (Kofler):184°C (with decomposition)
4-{3-[3-(2-Fluoro-4-trifluoromethylphenyl)ureido]-5-fluorobenzylamino}-
1 H-pyrazole-3-carboxamide

A solution of 22.3 g (36.89 mmol) of 4-{3-[3-(2-fluoro-4-
trifluoromethylphenyl)ureido]-5-fluorobenzylamino}-1H-pyrazole-3-(2,4-
dimethoxybenzylamide) and 17.54g (92.22 mmol) of para-toluenesulphonic acid in
600 ml of toluene is heated at reflux for 14 hours. After settling, the toluene solution is
separated from a yellow gum. The gum is stirred for 2 hours in 280 ml of water and
100 ml of sodium hydroxide (10 N). The suspension is filtered. The solid obtained is
rinsed with 3 x 350 ml of water then dried in order to give a cream solid which is
purified over 350 g of silica, eluted with a 95/2.5/2.5 (by volume) solution of
dichloromethane/methanol/acetonitrile: 3.85 g of 4-{3-[3-(2-fluoro-4-i
trifluoromethylphenyl)ureido]-5-fluorobenzylamino}-1H-pyrazole-3-carboxamide are!
obtained in the form of a white solid.
>H NMR (400 MHz, DMSO-Je) δ ppm 4.18 (d, J=5.9 Hz, 2 H) δ.68 - 5.77 (m, 1 H)
6.80 (d, J=8.8 Hz, 1H) 7.01 - 7.08 (br. s., 1 H) 7.04 (s, 1 H) 7.06 (s, 1 H) 7.24 (br. s., 1 H) 7.40
(dt, J=11.4, 2.0 Hz, 1 H) 7.54 (d, J=8.3 Hz, 1 H) 7.69 (d, J=11.5 Hz, 1 H) 8.41 (t, J=8.4 Hz, 1
H) 8.92 (br. s., 1 H) 9.40 (br. s., 1 H) 12.55 (br. s., 1 H)

MS: Retention time Tr (min) = 4.09; [M+H]+ m/z = 455; [M-H]- m/z = 453
Melting point (Kofler): 233°C
4-{3-[3-(2-Fluoro-4-trifluoromethylphenyl)ureido]-5-fluorobenzylamino}-
1 H-pyrazole-3-(2,4-dimethoxybenzylamide)

A solution of 11.40 g (36.45 mmol) of 4-amino-1 H-pyrazole-3-(2,4-
dimethoxybenzylamide) hydrochloride and of 6.65 ml (40.09 mmol) of
diisopropylethylamine in 360 ml of tetrahydrofuran is stirred at ambient temperature!
under an argon atmosphere. 4.35 g (36.45 mmol) of magnesium sulphate and 13.80 gj
(35.2 mmol) of 1-(2-fluoro-4-trifluoromethylphenyl)-3-(3-fluoro-5-formylphenyl)urea are!
added. The reaction mixture is then heated at reflux for 14 hours. The mixture is then
cooled to 25°C, then 11.46 g (182.25 mmol) of sodium cyanoborohydride are added
slowly. After stirring for 72 hours at ambient temperature, the mixture is concentrated
to dryness using a rotary evaporator. The gum obtained is stirred with 400 ml of water}
and 500 ml of a solution of sodium hydroxide (1N). This suspension is stirred fori
1 hour, then filtered through No. 3 sintered glass, the solid obtained is rinsed with
3 x 500 ml of water, then dried in an oven under vacuum at 40°C in order to give
22.45 g of 4-{3-[3-(2-fluoro-4-trifluoromethylphenyl)ureido]-5-fluorobenzylamino}-1 H-
pyrazole-3-(2,4-dimethoxybenzylamide) in the form of a pinkish solid.
'H NMR (500 MHz, DMSO-de) δ ppm 3.73 (s, 3 H) 3.81 (s, 3 H) 4.18 (d, J=6.3 Hz, 2
H) 4.33 (s, 2 H)-5.70 (t, J=6.2 Hz, 1 H) 6.47 (d, J=8.2 Hz, 1 H) 6.55 (s, 1 H) 6.79 (d, J=9.1 Hzl
1 H) 7.05 (s, 1 H) 7.07 - 7.12 (m, 2 H) 7.41 (d, J=11.3 Hz, 1 H) 7.53 (d, J=8.2 Hz, 1 H) 7.68 (d,
J=11.3 Hz, 1 H) 7.95 (br. s., 1 H)-8.40 (t, J=8.2 Hz, 1 H) 9.25 (br. s., 2 H) 12.52 (br. s., 1 H)
MS: Retention time Tr (min) = 4.79; [M+H>m/z = 605; [M-H]-m/z = 603

Melting point (Kofler):152°C
1-(2-Fluoro-4-trifluoromethylphenyl)-3-(3-f]uoro-5-formylphenyl)urea

A solution of 11.90 g (34.87 mmol) of 1-(2-fluoro-4-trifluoromethylphenyl)-3-(3-
cyano-5-fluorophenyl)urea in 150 ml of tetrahydrofuran is stirred at -2°C under an
argon atmosphere. Next, 86 ml of a solution of diisobutylaluminium hydride at a
concentration of 20% in hexane are added with a regular "dropwise" action. The
reaction mixture is stirred at ambient temperature for 12 hours. Next, an additional
60 ml of diisobutylaluminium hydride are added at -2°C. After stirring for 3 hours at
ambient temperature, the reaction medium is concentrated to dryness using a rotary
evaporator in order to give a thick oil to which 500 g of ice and 300 ml of 100% acetic
acid are slowly added, while stirring. The suspension obtained is filtered. The solid
obtained is washed with 4 x 150 ml of water, centrifuged and dried in an oven under
vacuum at 40°C in order to give 13.95 g of 1-(2-fluoro-4-trifluoromethylphenyl)-3-(3-
fluoro-5-formylphenyl)urea in the form of a pale yellow solid.
'H NMR (400 MHz, DMSO-de) 8ppm 7.33 - 7.38 (m, 1 H) 7.57 (d, J=8.8 Hz, 1 H)l
7.69 - 7.75 (m, 2 H)-7.79 (s, 1 H) 8.41 (t, J=8.3 Hz, 1 H) 9.05 (br. s., 1 H) 9.65 (s, 1 H) 9.98 (d,i
J=1.7 Hz, 1H)
MS: Retention time Tr (min) = 4.62; [M+H]+ m/z = 345; [M-H]- m/z = 343
Melting point (Kofler): 247°C
1-(2-Fluoro-4-trifluoromethylphenyl)-3-(3-cyano-5-fluorophenyl)urea


A solution of 6.15 g (45.18 mmol) of 5-fluoro-3-cyanoaniline in 90 ml of
tetrahydrofuran is stirred at ambient temperature under an argon atmosphere. 4.3 ml
(36.14 mmol) of diphosgene are added dropwise then 30 ml (135.54 mmol) of
triethylamine. After refluxing in the reaction mixture for 3 hours, a solution of 7.10 g
(39.64 mmol) of 4-amino-3-fluorothfluoromethylbenzene in 10 ml of tetrahydrofuran is
slowly added. The reflux is maintained for an additional 2 hours. The medium is then
stirred into 100 ml of water, then extracted with 100 ml of ethyl acetate. The organic
phase is washed with 100 ml of a saturated sodium chloride solution, dried over
magnesium sulphate and concentrated to dryness using a rotary evaporator. The solid
residue obtained is recrystallized at high temperature in 90 ml of acetonitrile in order
to give 10.35g of 1-(2-fluoro-4-trifluoromethylphenyl)-3-(3-cyano-5-fluorophenyl)urea
in the form of a cream solid.
'H NMR (400 MHz, DMSO-de) δ ppm 7.47 (d, J=8.3 Hz, 1 H) 7.57 (d, J=8.6 Hz, 1 H)|
7.69 (s, 1 H) 7.70 - 7.75 (m, 2 H) 8.37 (t, J=8.4 Hz, 1 H) 9.17 (br. s., 1 H) 9.63 (br. s., 1 H)
MS: Retention time Tr (min) = 1.1; [M+H]+: m/z 341.
Melting point (Kofler): 253°C
The products of the invention are useful as inhibitors of one or more reactions
catalysed by a kinase. KDR and/or Tie2 are kinases for which the products of the
invention will be particularly useful as inhibitors.
The reasons for which these kinases are chosen are given below.
KDR
KDR (Kinase insert Domain Receptor), also known as VEGF-R2 (Vascular
Endothelial Growth Factor Receptor 2), is mainly expressed in endothelial cells. This
receptor binds the proangiogenic growth factor VEGF, and thus serves as a
transduction signal mediator via the activation of its intracellular kinase domain. The
direct inhibition of the kinase activity of VEGF-R2 makes it possible to reduce the
phenomenon of angiogenesis in the presence of exogenous VEGF (Vascular

Endothelial Growth Factor) (Strawn ef al., Cancer Research, 1996, vol. 56, p.3540>
3545). This process has especially been demonstrated using VEGF-R2 mutants
(Millauer ef a/., Cancer Research, 1996, vol. 56, p.1615-1620). The VEGF-R2 receptor .
appears to have no other function in adults than that associated with the angiogenic
activity of VEGF. In addition to this central role in the dynamic angiogenic process,
recent results suggest that the expression of VEGF contributes towards the survival cjf
tumour cells after chemotherapy and radiotherapy, underlining the potential synergy of
KDR inhibitors with other agents (Lee et al. Cancer Research, 2000, vol. 60, p.5565-
5570).
Tie2
Tie-2 (TEK) is a member of a family of tyrosine kinase receptors, which is
specific to endothelial cells. Tie2 is the first receptor with tyrosine kinase activity for
which both the agonist (angiopoietin 1 or Ang1), which stimulates trie
autophosphorylation of the receptor and cell signalling [S. Davis et al. (1996) Cell 8:7,
1161-1169], and the antagonist (angiopoietin 2 or Ang2) [P.C. Maisonpierre et al. (1997)
Science 277, 55-60] are known. Angiopoietin 1 can have a synergistic effect with VE0F
in the final stages of neoangiogenesis [Asahara T., Circ. Res. (1998) 233-240]. Knock-
out experiments and transgenic manipulations of the expression of Tie2 or of Ang1 leid
to animals that present vascularization defects [D.J. Dumont et al. (1994) Genes Dev. |8,
1897-1909 and C. Suri (1996) Cell 87, 1171-1180]. The binding of Ang1 to its receptor
leads to autophosphorylation of the kinase domain of Tie2, which is essential for
neovascularization and also for the recruitment and interaction of blood vessels with the
pericytes and smooth muscle cells; these phenomena contribute towards the maturation
and stability of the newly formed blood vessels [P.C. Maisonpierre ef al. (1997) Scienpe
277, 55-60]. Lin et al. (1997) J. Clin. Invest. 100, 8: 2072-2078 and Lin P. (1998) PNAS
95, 8829-8834 have shown an inhibition of tumour growth and vascularization, and albo
a reduction in lung metastases, during adenoviral infections or injections of the
extracellular domain of Tie-2 (Tek) into models of melanoma and breast tumour
xenografts.
For the reasons that follow, the Tie2 inhibitors may be used in situations in which
neovascularization or angiogenesis takes place inappropriately, i.e. in cancers: in
general, but also in particular cancers such as Kaposi's sarcoma or infantile
haemoangioma, rheumatoid arthritis, osteoarthritis and/or its associated pain,
inflammatory diseases of the intestine such as haemorrhagic rectocolitis or Crohn's
disease, eye pathologies such as age-related macular degeneration, diabetic
retinopathies, chronic inflammation and psoriasis.

Angiogenesis is a process of generation of new blood capillaries from prel
existing blood vessels. Tumour angiogenesis (formation of new blood vessels), which i$
essential for tumour growth, is also one of the essential factors of metastasic
dissemination (Oncogene. 2003 May 19; 22(20):3172-9; Nat. Med. 1995 Jan; 1(1):27f
31).
This neovascularization is due to the migration and then the proliferation and
differentiation of endothelial cells under the influence of angiogenic factors secreted by
cancer cells and stromal cells (Recent Prog. Horm. Res. 2000; 55:15-35; 35-6).
The angiopoietin 1/Tie2 receptor system plays a predominant role in the
maturation of blood vessels by allowing the recruitment of periendothelial cells to
stabilize the vessels (Cell. 1996 Dec. 27; 87(7): 1161-9, Recent Prog. Horm. Res. 2004;
59:51-71). Thus, it has been shown that the administration of the soluble recombinant
form of the extracellular domain of the Tie-2 receptor (exTek) inhibits tumoijir
angiogenesis in models of murine tumours, and also metastatic growth (Pengnian Lin,
Jake A. Buxton, Ann Acheson, Czeslaw Radziejewski, Peter C. Maisonpierre, George
D. Yancopoulos, Keith M. Channon, Laura P. Hale, Mark W. Dewhirst, Samuel E.
George and Kevin G. Peters, Proc. Natl Acad. Sci. USA. 1998 Jul 21; 95(15): 8829-34;
Cecilia Melani, Antonella Stoppacciaro, Chiara Foroni, Federica Felicetti, Alessandra
Care and Mario P. Colombo, Cancer Immunol Immunother. 2004 Jul; 53(7): 600-8). In
endothelial cells in culture, stimulation of Tie-2 activates the PI3 kinase pathway, of p42/
p44 pathways involved in cell proliferation and migration; of the synthesis of PAF (Cell
Signal. 2006 Apr 14; ahead of print), a pathway involved in pro-inflammatory activity.
Stimulation of Tie2 stimulates the Akt pathway and inhibits apoptosis (Laura M DeBusk,
Dennis E Hallahan, Pengnian Charles Lin, Exp. Cell Res. 2004 Aug. 1; 298(1): 167-77),
a transduction pathway known for its importance in cell survival.
The addition of exTek (soluble receptor of Tie2) inhibits the formation lof
pseudotubules of endothelial cells on Matrigel (Cecilia Melani, Antonella Stoppacciaro,
Chiara Foroni, Federica Felicetti, Alessandra Care and Mario P. Colombo, Cancer
Immunol Immunother. 2004 Jul; 53(7): 600-8). These studies indicate that the Tie-
2/angiopoietin system is necessary during the first stages of formation of vascular buds
in adult tissues and that one function of the Tie-2 receptor is to increase the survival! of
endothelial cells during the formation of blood vessels. Furthermore, angiopoietin-1
stimulates the proliferation of lymphatic endothelial cells and also lymphangiogenesis
(development of new lymphatic vessels), a favoured access pathway for metastatic
growth (Tohru Morisada, Yuichi Oike, Yoshihiro Yamada, Takashi Urano, Masaki Akao,
Yoshiaki, Kubota, Hiromitsu Maekawa, Yoshishige Kimura, Masako Ohmura, Takeshi
Miyamoto, Shiro Nozawa, Gou Young Koh, Kari Alitalo and Toshio Suda, Blood. 2005

Jun 15; 105(12): 4649-56).
Among the enzymes that have a role in angiogenesis, mention may also be
made of PDGFR(3 (Guzman and Laurence H. Hurley, Univ. of Arizona, Molecular
Cloning of the Human PDGFR-beta Promoter and Targeting the G-Quadruplex-
Forming Region to Control Gene Expression; Biomedical Drug Discovery, Genomics/
Gentics, Therapeutic), FGFR1 (Somaia Elbauomy Elsheikh, Andrew R Green,1
Maryou BK Lambros, Nicholas C Turner, Matthew J Grainge,3 Des Powe,1 Ian O Ellis,
and Jorge S Reis-Filho, FGFR1 amplification in breast carcinomas: a chromogenic in
situ hybridization analysis; FLT1 (Shibuya M (2007), "Vascular endothelial growth
factor receptor-1 (VEGFR-1/FIM): a dual regulator for angiogenesis", Angiogenesis, 9
(4): 225-30; discussion 231 and VEGFR3 (Tamela, T. et al., Blocking VEGFR-3
suppresses angiogenic sprouting and vascular network formation, Nature 454, 656-
660 (20058). j
Angiogenesis processes also play a predominant role in the progression pf
numerous solid tumours. Furthermore, it has been shown that the probability of onset ft
metastases increases very greatly as the vascularization of the primary tumojjr
i
increases (Br. J. Cancer. 2002 May 20; 86(10): 1566-77).
The potential role of proangiogenic agents in leukaemias and lymphomas has also mofe
recently been documented. Specifically, it has been reported in general that cell clones
in these pathologies may be either naturally destroyed by the immune system, or revert
to an angiogenic phenotype that favours their survival and then their proliferation. This
change in phenotype is induced by an overexpression of angiogenic factors especially
by the macrophages and/or mobilization of these factors from the extracellular matrix
(Thomas DA, Giles FJ, Cortes J, Albitar M, Kantarjian HM., Acta Haematol., (2001), vol.
207, pp. 106-190).
There is a correlation between the angiogenesis process of bone marrow afid
"extramedullar diseases" in CML (chronic myelomonocytic leukaemia). Various studies
demonstrate that the inhibition of angiogenesis might represent a treatment of choice in
this pathology (Leuk. Res. 2006 Jan; 30(1): 54-9; Histol. Histopathol. 2004 Oct.; 19(|4):
1245-60). Furthermore, it is strongly suggested that activation of the Tie2/angiopoie|tin
system is involved in the development of angiogenesis of bone marrow in the case! of
patients suffering from multiple myeloma (Blood, 2003 Jul 15; 102(2): 638-45).

Determination of the activity of the compounds - experimental protocols
1. KDR
The inhibitory effect of the compounds is determined in an in vitro test Of
phosphorylation of substrate via a scintillation technique (96-well plate of basic Flasjn
Plate type).
The cytoplasmic domain (residues 790 to 1356) of the human KDR enzyme was
cloned in the form of a GST fusion in the pFastBac baculovirus expression vector. Thb
protein was expressed in the SF21 cells, purified and activated by autophosphorylatioh.
The substrate is composed of residues 658 to 850 of the PLCy expressed and purified
in the form of a GST fusion protein.
The KDR kinase activity is measured in the buffer 20mM MOPS, 10mM MgCU,
10mM MnCI2, 1mM DTT, pH = 7.4. The compounds are initially diluted in 100% DMSO,
then prepared as a 10X solution in 30% DMSO/70% buffer. 10 ul of the 10X solution arte
deposited, then 70 ul of buffer containing 150 ng (1.6 pmol) of KDR enzyme at 4°C. The
reaction is initiated by adding 20 u.l of solution containing 2 ug (41 pmol) of PLCty
substrate, 0.5 uCi of f3P[ATP] and 2uM of cold ATP. The plate is agitated. Aft0r
incubation for 30 minutes at 37°C, the incubation buffer is removed, and the wells are
washed three times with 300 [x\ of PBS. The radioactivity in each well is measured usirjg
a Trilux-BWallac radioactivity counter.
The background noise is determined by measuring the radioactivity in four
different wells containing ATP (radiolabeled and cold) and the substrate, in the absence
of enzyme and of compound. The control activity is measured in four different wells
containing all the reagents, but in the absence of compound.
The inhibition of the KDR activity with the compound of the invention is
expressed as a percentage of inhibition of the control activity determined in the absence
of compound.
2. Tie2
The inhibitory effect of the compounds is determined in an in vitro test of
phosphorylation of substrate via a scintillation technique (96-well plate of basic Flash
Plate type).
The coding sequence of human Tie2 corresponding to the amino acids of the
intracellular domain 774-1124 was introduced into a pFastBacGT baculovirus
expression vector in the form of a GST fusion protein. GST-Tie2 was purified and

activated by autophosphorylation. The substrate is composed of residues 658 to 850 pf
the PLCy expressed and purified in the form of a GST fusion protein.
The kinase activity of Tie2 is measured in a MOPS 20mM pH 7.4 buffer,
containing 10mM MgCI2, 10mM MnCI2 and 1mM DTT. The compounds are initial y
diiuted in 100% DMSO, then prepared as a 10X solution in 30% DMSO/70% buffer. 10
uJ of the 10X solution are deposited, then 70 ul of buffer containing 100 ng (1.5 pmol) 0.040 ml.hMO"6 cell: high intrinsic clearance
Demographic information on the donor:
Preparations of cryopreserved human hepatocytes:
From IVT: • IVT, TLN group: man, Caucasian, 25 years old
Preparations of fresh human hepatocytes:
From Biopredic: • HEP200239: woman, unknown, 58 years old
In this test, the intrinsic clearance value of the compound described in example
1 is 0.057 ml/hour/million of cells (value classified as intermediate with a low inter-
individual variability).
The intrinsic clearance value of the compound described in example 2 is
0.055 ml/h/million of cells (value classified as intermediate with a low inter-individual
variability).

[0024] Other tests consisting in measuring the in vivo activity of the compounds of the
invention on colon tumours were carried out.
[0025] This activity of the compounds of the invention on colon tumours was studied
on the B16 melanoma. In comparison, the product from example 19 of application
WO 08/065282 was tested.
The effectiveness of a product can be determined in vivo by various criteria, it
is possible to determine it by the percentage of tumour inhibition %T/C, which
represents the ratio between the average weight of the tumours of the treated group
(T) and the average weight of the tumours of the control group (C) on day 12 or 13 of
treatment. A product is considered to be active when the T/C ratio is less than 42%
and a product has a high antitumour activity when the T/C is less than 10%. (Corbett
TH et al., Cancer Research, 42, 1707-1715 (1982).
In order to demonstrate the effectiveness of a compound, it is also possible to
determine the logio cell kill, which is determined according to the following equation:
logio cell kill = T-C (days)/3.32 x Td
in which T-C represents the delay in growth of the cells, which is the average time, in
days, for the tumours of the treated group (T) and the tumours of the control group (C)
to have reached a predetermined value (750 mg for example), and Td represents the
time, in days, necessary for the volume of the tumour to double in the control animals
[T.H. Corbett et al., Cancer, 40, 2660.2680 (1977); F.M. Schabel et al., Cancer Drug
Development, Part B, Methods in Cancer Research, 17, 3-51, New York, Academic
Press Inc. (1979)]. A product is considered to be active if logio cell kill is greater than
or equal to 0.7. A product is considered to be very active if logio cell kill is greater than
2.8.
The effectiveness of the compounds on solid tumours can be determined
experimentally in the following way:
The animals subjected to the experiment, generally C57BL/6 female mice,
are grafted bilaterally, subcutaneously, with 30 to 60 mg of a B16 (reference of the
tumour) human tumour fragment on day 0. The animals bearing the tumours are
randomized before being subjected to the various treatments and controls. In the case
of treatment of tumours of the present invention, the treatment was started at an early
stage, 3 to 4 days after implantation. The animals which underwent the treatment with
the compounds had a weight of around 20 g. Animals bearing tumours were also
subjected to the same treatments with the excipient alone in order to be able to

dissociate the toxic effect of the excipient from the actual effect of the chemotherapy!
on the tumour. The administrations of the compounds were carried out orally at the
doses indicated in the tables and with the excipients indicated in the tables, according
to a daily double administration. These administrations were carried out over 8 to 11
days, depending on the study, after implantation of the tumour.
The tumours are measured two or three times a week until the tumour!
reaches approximately 2 g or until the death of the animal if the latter occurs before
the tumour reaches 2 g. The animals are autopsied at the time of sacrifice.
The antitumour activity is determined according to the various parameters
recorded.
By way of examples, the following tables give the results obtained with the
compounds of the invention used at their optimum dose.
[0027] Thus, one subject of the invention is medicaments that comprise a compound
of formula (I), or an addition salt of the latter with a pharmaceutically acceptable acid.
[0028] According to another of its aspects, the invention relates to compounds
corresponding to formula (I) for the use thereof as a medicament in the treatment of
cancer. The compounds according to the invention may be used for the preparation of
medicaments, in particular medicaments that inhibit angiogenesis.
[0029] These medicaments find their therapeutic use especially in the treatment of;
cancerous tumours, especially solid tumours.
[0030] According to another of its aspects, the present invention relates to!
pharmaceutical compositions containing, as active principle, a compound according to
the invention. These pharmaceutical compositions contain an effective dose of at least:
one compound according to the invention, or a pharmaceutically acceptable salt of;
said compound, and also at least one pharmaceutically acceptable excipient.
Said excipients are chosen according to the pharmaceutical form and the
desired mode of administration, from the usual excipients known to those skilled in the
art.
[0031] In the pharmaceutical compositions of the present invention for oral,

sublingual, subcutaneous, intramuscular, intravenous, topical, local, intratracheal,
intranasal, transdermal or rectal administration, the active principle of formula (1)
above, or the salt thereof, may be administered in a unit administration form, as a
mixture with standard pharmaceutical excipients, to animals and to human beings for
the treatment of the above disorders or diseases.
[0032] The appropriate unit administration forms include oral forms such as tablets,
soft or hard gel capsules, powders, granules and oral solutions or suspensions,
sublingual or buccal administration forms and transdermal, subcutaneous,
intramuscular or intravenous administration forms.
[0033] By way of example, a unit administration form of a compound according to the;
invention in the form of a tablet may comprise the following components:
Compound according to the invention 50.0 mg
Mannitol 223.75 mg
Croscaramellose sodium 6.0 mg
Cornstarch 15.0 mg
Hydroxypropyl methyl cellulose 2.25 mg
Magnesium stearate 3.0 mg
[0034] When given orally, the dose of active principle administered per day can reach
1200 mg/kg, in one or more intakes.
[0035] There may be particular cases where higher or lower dosages are appropriate;
such dosages do not depart from the context of the invention. Depending on the usuail
practice, the dosage appropriate for each patient is determined by the physician
depending on the method of administration and the weight and response of said
patient.
[0036] According to another of its aspects, the present invention also relates to a
method for treating the pathologies indicated above, which comprises the
administration, to a patient, of an effective dose of a compound according to the
invention, or a pharmaceutically acceptable salt thereof.

CLAIMS
1. Compound corresponding to the formula (I):

in which:
X represents chlorine or fluorine,
in the form of base or acid-addition salt.
2. Compound of formula (I) according to Claim 1, characterized in that X
represents chlorine, in the form of base or of acid-addition salt.
3. Process for preparing a compound of formula (I) according to either one of
Claims 1 and 2, characterized in that the following compound:

is reacted with the compound:

in the presence of diisopropylethylamine in an inert medium, then in a second step, the
amine is deprotected by para-toluenesulphonic acid.
4. Process according to Claim 3, characterized in that the inert medium is an
apolar aprotic medium, preferably tetrahydrofuran.

5. Compound of formula:

where X represents chlorine or fluorine.
6. Compound of formula:

where X represents chlorine or fluorine.
7. Medicament, characterized in that it comprises a compound of formula (1)
according to either one of Claims 1 and 2, or an addition salt of this compound with a
pharmaceutically acceptable acid.
8. Pharmaceutical composition, characterized in that it comprises a compound of
formula (I) according to either one of Claims 1 and 2, or a pharmaceutically acceptably
salt of this compound, and also at least one pharmaceutically acceptable excipient.
9. Compound corresponding to the formula (I) according to either one of Claims fl
and 2 for its application as a medicament in the treatment of cancer.

ABSTRACT

The invention relates to pyrazole derivatives of the general formula (I), where X is chlorine or fluorine. The
invention also relates to a method for preparing same and to the therapeutic use thereof.