FLUOROPHENYL PYRAZOL COMPOUNDS
This invention relates to compounds or pharmaceutically acceptable salts thereof,
and therapeutic use thereof. Compounds of this invention are inhibitors of methionine
aminopeptidase 2 (MetAP2).
Obesity is a complex medical disorder resulting in excessive accumulation of
adipose tissue mass. Today obesity is a world-wide public health concern that is
associated with cardiovascular disease, diabetes, certain cancers, osteoarthritis, and other
undesired health outcomes. The treatment of obesity strives to reduce excess body
weight, improve obesity-related morbidity, and maintain long-term weight reduction.
Approved drugs for treating obesity offer particularly unsatisfactory efficacy for severely
obese subjects. There is a need for alternative treatment options to induce desired weight
loss in a patient.
Increased expression of the MetAP-2 gene has been historically associated with
various forms of cancer; however, WO 2010/065879 reports molecules inhibiting the
enzymatic activity of MetAP-2 for use in treating obesity. There is a need for novel
inhibitors of MetAP-2 for use in treating a condition associated with MetAP-2 increased
expression.
The present invention provides novel compounds which are MetAP2 inhibitors.
MetAP2 inhibitor compounds are desired to provide treatments for MetAP2 mediated
conditions, such as obesity.
The present invention provides a compound of the Formula I below:
wherein X is selected from the group consisting
R is selected from the group consisting of H and ¾ ;
R is selected from the group consisting of H, CH , F, CI, OCH , C(0)OH, C(0)NH 2 and
or a pharmaceutically acceptable salt thereof.
In an embodiment of the invention, X is selected from the group consisting of
In an embodiment of the invention X is . In another embodiment of
the invention, X is
In an embodiment of the invention R is selected from the group consisting of H,
F, and CH3. In an embodiment R is . In an embodiment of the invention R
is CH .
In a preferred embodiment, the compound is is [3-[[3-(4-Fluorophenyl)-lHpyrazol-
4-yl]oxy]phenyl]methylurea, or a pharmaceutically acceptable salt thereof.In a
preferred embodiment the compound is [6-[[3-(4-Fluorophenyl)-lH-pyrazol-4-yl]oxy]-5-
methyl-2-pyridyl]methylurea or a pharmaceutically acceptable salt thereof.
The present invention also provides a pharmaceutical composition comprising a
compound of Formula I as described above or a pharmaceutically acceptable salt thereof
together with one or more pharmaceutically acceptable carriers, diluents or excipients.
The present invention also provides a method for treating obesity in a mammal.
The method comprises administering to the mammal in need of treatment a compound as
described above for Formula I, or a pharmaceutically acceptable salt thereof. The
invention provides a method for inducing desired weight loss in a mammal in need
thereof, comprising administering an effective amount of a compound of Formula I . The
invention provides a method for therapeutic weight weight loss in a mammal in need
thereof, comprising administering an effective amount of a compound of Formula I .
The present invention provides a compound according to Formula I or a
pharmaceutically acceptable salt thereof as described above for use in therapy.
In yet another form, the present invention provides a compound as described
above according to Formula I, a pharmaceutically acceptable salt thereof, or
pharmaceutical composition for use in the treatment of obesity in a mammal in need
thereof. Preferably the mammal is a human.
The present invention provides use of a compound according to Formula I, or a
pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the
treatment of obesity. The present invention provides the use of a compound according to
Formula I, or a pharmaceutically acceptable salt thereof, for use in the manufacture of a
medicament for use in providing therapeutic weight loss.
Compounds of the present invention can be provided as a pharmaceutically
acceptable salt. "Pharmaceutically-acceptable salt" refers to salts of the compound of the
invention considered to be acceptable for clinical and/or veterinary use. Pharmaceutically
acceptable salts and common methodology for preparing them are well known in the art.
See, e.g., P. Stahl, et a , Handbook of Pharmaceutical Salts: Properties, Selection and
Use, (VCHA/Wiley-VCH, 2002); S.M. Berge, etal, "Pharmaceutical Salts," Journal of
Pharmaceutical Sciences, Vol. 66, No. 1, January 1977.
The abbreviations used herein are defined according to Aldrichimica Acta, Vol.
17, No. 1, 1984. Other abbreviations are defined as follows: "BSA" refers to Bovine
Serum Albumin; "DMF' refers to ,-dimethylformamide; "DIO" refers to diet induced
obese; "HEC" refers to hydroxy ethyl cellulose; "HEPES" refers to 4-(2-hydroxyethyl)-lpiperazineethanesulfonic
acid; "HFD" refers to high fat diet; "IC50" refers to the
concentration of an agent that produces 50% of the maximal inhibitory response possible
for that agent; and "THF" refers to tetrahydrofuran.
The intermediates described in the following Schemes and preparations may
contain a number of nitrogen, hydroxy, and acid protecting groups such as esters. The
variable protecting group may be the same or different in each occurrence depending on
the particular reaction conditions and the particular transformations to be performed. The
protection and deprotection conditions are well known to the skilled artisan and are
described in the literature. See. e.g., Greene and Wuts, Protective Groups in Organic
Synthesis, (T. Greene and P. Wuts, eds., 2d ed. 1991).
In the schemes below, all substituents unless otherwise indicated, are as
previously defined. The reagents and starting materials are generally readily available to
one of ordinary skill in the art. Others may be made by standard techniques of organic
and heterocyclic chemistry which are analogous to the syntheses of known structurallysimilar
compounds and the procedures described in the Preparations and Examples which
follow including any novel procedures.
A compound of Formula I can be prepared in accordance with reactions as
depicted in Scheme 1. Scheme 1 (Step 1) depicts the alkylation of a 2-bromo-l-(4-
fluorophenyl)ethanone (1) with a substituted aryl or pyridyl hydroxy nitrile (2) as denoted
by X with X defined previously. The bromo (1) reagent is reacted with the aryl or
heteroaryl hydroxy compound (2) under basic conditions well known in the art using an
inorganic base such as potassium carbonate in a polar aprotic solvent such as acetonitrile
or acetone at room temperature or refluxing conditions to give compound 3, Step 1.
Compound 3 is alkylated with ,-dimethylformamide dimethyl acetal in a non-polar
solvent such as toluene under refluxing conditions or microwave irradiation to give
compound 4, Step 2, the substituted dimethylamino prop-2-ene-l-one compound (4).
Compound 4 can be isolated or carried on in-situ to Step 3 to form the pyrazole
compound (5). The pyrazole can be formed with hydrazine hydrate in a non-polar solvent
such as toluene under microwave irradiation or alternatively with hydrazine
hydrochloride using an organic base such as triethylamine in a polar protic solvent such
as ethanol at refluxing conditions to give the substituted cyclized pyrazole (5). For A =
nitrile, the nitrile on the pyridyl or phenyl (5) can be reduced to the amine (6) using
reducing conditions well known in the art such as lithium aluminum hydride in a polar
aprotic solvent such as THF or under hydrogen conditions using Raney nickel in a
methanol ammonia solution to give the amine (6, Step 4). The methyl urea compounds of
Formula I can be formed from the amine (6) using phenylurethane in a polar aprotic
solvent such as THF and an organic base such as diisopropylethylamine while heating to
about 80 °C under microwave irradiation conditions to give compounds of Formula I
(Step 5). Alternatively, an inorganic base such as potassium cyanide and an acid such as
acetic acid in a polar protic solvent such as methanol with heating to about 80 °C under
microwave irradiation conditions can be used to convert the amine (6) to the methyl urea
compounds of Formula I (Step 5). For A = Br, the bromide can be converted to the nitrile
(7, Step 6) under palladium conditions well known in the art using ZnCN and a
palladium(O) catalyst such as [l,l'-bis(diphenylphosphino)
ferrocene]dichloropalladium (II) with zinc and zinc acetate in dimethylacetamide with
heating to about 160 °C to give the nitrile compound (7, Step 6) which can be carried on
to compounds of Formula I as described above for Steps 4 and 5. Alternatively, the
nitrile (7) can be formed using copper cyanide and a base such as N-methyl pyrrolidine
and heating to about 200 °C followed by the addition of ethylene diamine to give
compound (7, Step 6).
Scheme 2
(2) For A = Br
Alternatively, in Scheme 2, for A = Br of compound 2, the bromide (2) can be
converted to a nitrile using copper cyanide in a polar solvent such as Nmethylpyrrolidone
and heated to about 200 °C under microwave irradiation to give a
nitrile. (8). This hydroxy nitrile can then be reacted with compound (1) in Scheme 1,
Step 1, as described in Scheme 1 and carried on to give compounds of Formula I as
described above for Scheme 1, Steps 2-5.
The following preparations and examples further illustrate the invention and
represent typical synthesis of the compound of Formula (I). Unless noted to the contrary,
the compounds illustrated herein are named and numbered using Accelrys Draw 4.0,
IUPACNAME ACDLABS or Symyx /Draw 4.0.
Preparation 1
4-Fluoro-3-hydroxy-benzonitrile
Add lithium chloride (1.7 g, 40 mmol) to a stirred solution of 4-fluoro-3-
methoxybenzonitrile (2.0 g, 13 mmol) in DMF (26 mL). Stir the resulting mixture at 160
°C for 5 hours and then cool to room temperature with stirring for 12 hours. Dilute the
mixture with ethyl acetate (50 mL) and wash with water (2 x 40 mL). Acidify the
isolated aqueous phase with 1M HCl and extract with ethyl acetate (3 x 60 mL). Wash
the combined organic extracts with water (2 x 100 mL), dry over Na2S04, and
concentrate under reduced pressure to give the title compound as a yellow solid (0.68 g,
38%): ES/MS (m/z) 138 (M+H), which is directly used without further purification.
Preparation 2
5-Hydroxy-2-methyl-benzonitrile
Add a solution of sodium nitrile (2.1 g, 3 1 mmol) in water (15 mL) drop wise at 5
°C to a stirred solution of 5-amino-2-methylbenzonitrile (4.0 g, 3 1 mmol) in 33% sulfuric
acid (45 mL) and keep the temperature below 5 °C. In a separate flask, add concentrated
sulfuric acid (30 mL) cautiously to a stirred solution of sodium sulfate (21.7 g, 153 mmol)
in water (15 mL) and heat the mixture to reflux. Add the prepared diazonium solution to
the refluxing mixture in portions and continue refluxing for 2 hours. Cool the mixture
slowly to room temperature and stir overnight. Extract the mixture with ethyl acetate (2 x
100 mL) and wash the combined organic extracts with water (2 x 150 mL) and 10%
NaOH water solution (3 x 100 mL). Acidify the combined NaOH extracts with
concentrated HC1 and then extract with ethyl acetate (2 x 200 mL). Dry the organic
extracts over Na2S0 4 and concentrate under reduced pressure to give the title compound
as a yellow solid (2.4 g, 60%): ES/MS (m/z) 134 (M+H), which is used directly without
further purification.
Preparation 3
3-Cyano-5-hydroxy-benzoic acid
Add copper cyanide (0.90 g, 10 mmol) to a stirred solution of 3-bromo-5-
hydroxybenzoic acid (2.2 g, 10 mmol) in N-methylpyrrolidone (12 mL). Heat the mixture
to 200 °C under microwave irradiation for 2 hours. Pour the mixture into 1M HC1 water
solution (100 mL) and extract with ethyl acetate (3 x 50 mL). Combine the organic
extracts, wash with brine, dry over Na2S0 4 and concentrate under reduced pressure to
give the title compound as a brown solid (1.5 g, 92%). ES/MS (m/z) 164 (M+H), which
is used directly without further purification.
Preparation 4
Methyl 3-cyano-5-hydroxy-benzoate
Add concentrated sulfuric acid (0.42 mL, 7.9 mmol) into a stirred solution of 3-
cyano-5 -hydroxy-benzoic acid (3.2 g, 19.6 mmol) in methanol (50 mL). Stir the mixture
at 70 °C for 20 hours and then stir at room temperature for 2 days. Evaporate the solvent
under reduced pressure and add saturated NaHCC>3 water solution (100 mL) into the
residue. Filter the precipitated solid and wash it with cold water (20 mL). Dissolve the
solid in ethyl acetate (100 mL), dry over Na2S0 4, and concentrate under reduced pressure
to give the title compound as a grey solid (2.9 g, 82%): ES/MS (m/z) 178 (M+H), which
is used directly without further purification.
Preparation 5
2-Cyano-5-methylpyridine 1-oxide
Add m-chloroperoxybenzoic acid (8.0 g, 46 mmol) in batches to a stirred solution
of 5-methylpicolinonitrile (4.0 g, 34 mmol) in dichloromethane (60 mL). Stir the mixture
under refluxing conditions for 24 hours and cool to room temperature. Wash the mixture
with saturated Na2S2 0 water solution (3 x 20 mL) and brine (3 x 20 mL), dry over
Na2S0 4 and concentrate under reduced pressure to give a residue. Purify the residue by
silica gel flash chromatography, eluting with a gradient solvent of 50-100% ethyl acetate
in petroleum ether to give the title compound as a yellow solid (4.2 g, 92%). ES/MS
(m/z) 135 (M+H).
The following compound is prepared essentially by the method of Preparation 5.
Table 1
ES/MS
Prep.
Chemical Name Structure (m/z)
No
(M+H)
0 11 2-Cyano-3- 6
methylpyridine 1-oxide 135
Preparation 7
-3-methyl-2-pyridyl) acetate
Heat a mixture of 2-cyano-5-methylpyridine 1-oxide (3.7 g, 28 mmol) in acetic
acid anhydride (40 mL) at 145 °C with stirring for 64 hours and cool to room
temperature. Evaporate the solvent under reduced pressure to give a residue and purify
the residue by silica gel flash chromatography, eluting with a gradient solvent of 0-50%
ethyl acetate in petroleum ether to give the title compound as a yellow solid (4.0 g, 82%)
ES/MS (m/z) 135 (M-C2H 0+H).
The following compound is prepared essentially by the method of Preparation 7.
Table 2
Preparation 9
6-Hydroxy-5-methyl-pyridine-2-carbonitrile
Add sodium hydroxide (2.1 g, 53 mmol) to a stirred solution of (6-cyano-3-
methyl-2-pyridyl) acetate (3.4 g, 19 mmol) in methanol (100 mL) and water (100 mL).
Stir the mixture at room temperature for 2 hours and evaporate the solvent under reduced
pressure. Add citric acid (5.6 g, 29 mmol) and partition the resulting mixture between
ethyl acetate (150 mL) and water (120 mL). Isolate the organic phase, wash with brine (3
x 50 mL), dry over Na2S04 and concentrate under reduced pressure to give the title
compound as a white solid (2.5 g, 97%), which is used directly without further
purification. ES/MS (m/z) 135 (M+H).
The following compound is prepared essentially by the method of Preparation 9.
Table 3
Preparation 11
3-[2-(4-Fluorophenyl)-2-oxo-ethoxy]benzonitrile
Add potassium carbonate (12.0 g, 86.8 mmol) to a stirred solution of 2-bromo-l-
(4-fluorophenyl)ethanone (9.0 g, 41.5 mmol) and 3-hydroxybenzonitrile (5.0 g, 42.0
mmol) in acetonitrile (50 mL). Stir the mixture under refluxing condition for 2 hours.
Filter off the solid and concentrate the filtrate under reduced pressure to give the title
compound as a white solid (10.8 g, 103% crude), which is used directly without further
purification. ES/MS (m/z) 256 (M+H).
The following compound is prepared essentially by the method of Preparation 11.
Table 4
ES/MS
Prep.
Chemical Name Structure (m/z)
No
(M+H)
3-Fluoro-5-[2-(4-
12 fluorophenyl)-2-oxo- 274
ethoxyjbenzonitrile
Preparation 13
4-Fluoro-3-[2-(4-fluorophenyl)-2-oxo-ethoxy]benzonitrile
Add potassium carbonate (1.0 g, 7.5 mmol) to a stirred solution of 2-bromo-l-(4-
fluorophenyl)ethanone (1.1 g, 5.0 mmol) and 4-fluoro-3-hydroxybenzonitrile (0.68 g, 5.0
mmol) in acetone (30 mL). Stir the mixture at room temperature for 1 hour. Filter off the
solid and concentrate the filtrate under reduced pressure to give a residue. Purify the
residue by silica gel flash chromatography, eluting with ethyl acetate: petroleum ether
(1:5) to give the title compound as a yellow solid (0.99 g, 73%). ES/MS (m/z) 274
(M+H).
The following compounds are prepared essentially by the method of Preparation
Table 5
Preparation 20
2-[(6-Bromo-2-pyridyl)oxy] -1-(4-fluorophenyl)ethanone
Add silver(I) carbonate (1.7 g, 6.2 mmol) to a stirred solution of 2-bromo-l-(4-
fluorophenyl)ethanone (0.87 g, 4.0 mmol) and 6-bromopyridin-2-ol (0.70 g, 4.0 mmol) in
toluene (10 mL). Heat the mixture to 105 °C under microwave irradiation with stirring
for 12 hours. Filter off the solid and concentrate the filtrate under reduced pressure to
give a residue. Purify the residue by silica gel flash chromatography, eluting with ethyl
acetate: hexanes (1:1) to give the title compound as a light yellow solid (0.74 g, 60%).
ES/MS (m/z) ( Br/ Br): 309.9/312.0 [M+H].
The following compound is prepared essentially by the method of Preparation 20.
Table 6
Preparation 22
2-[(5-Bromo-3-pyridyl)oxy]-l-(4-fluorophenyl)ethanone
Add potassium carbonate (8.0 g, 57 mmol) to a stirred solution of 2-bromo-l-(4-
fluorophenyl)ethanone (6.2 g, 29 mmol) and 3-bromo-5-hydroxypyridine (5.0 g, 29
mmol) in acetone (100 mL). Stir the mixture at room temperature for 1 hour. Filter off
the solid and concentrate the filtrate under reduced pressure to give a residue. Purify the
residue by silica gel flash chromatography, eluting with a gradient solvent of 10-30%
ethyl acetate in petroleum ether to give the title compound as a yellow solid (3.6 g, 40%).
ES/MS (m/z) ( Br/ Br): 309.8/311.8 [M+H].
The following compound is prepared essentially by the method of Preparation
Table 7
Preparation 24
6-[2-(4-Fluorophenyl)-2-oxo-ethoxy]-5-methyl-pyridine-2-carbonitrile
Add potassium carbonate (5.0 g, 36 mmol) to a stirred solution of 2-bromo-l-(4-
fluorophenyl)ethanone (3.9 g, 18 mmol) and 6-hydroxy-5-methyl-pyridine-2-carbonitrile
(2.4 g, 18 mmol) in acetone (100 mL). Stir the mixture at room temperature for 2 hours
and partition between ethyl acetate (150 mL) and water (70 mL). Isolate the organic
layer, wash with brine (3 x 60 mL), dry over Na2S04 and concentrate under reduced
pressure to give a residue. Purify the residue by silica gel flash chromatography, eluting
with dichloromethane to give the title compound as a white solid (4.0 g, 79%). ES/MS
(m/z) 271 (M+H).
The following compound is prepared essentially by the method of Preparation 24.
Table 8
Preparation 26
(E)-2-[(6-Bromo-2-pyridyl)oxy]-3-(dimethylamino)-l-(4-fluorophenyl)prop-2-en-l-one
Add N,N -dimethylformamide dimethyl acetal (0.30 g, 2.5 mmol) to a stirred
solution of 2-[(6-bromo-2-pyridyl)oxy]-l-(4-fluorophenyl)ethanone (0.70 g, 2.3 mmol) in
toluene (8 mL). Heat the mixture to 120 °C under microwave irradiation with stirring for
3 h and evaporate the solvent under reduced pressure to give a residue. Purify the residue
by silica gel flash chromatography, eluting with ethyl acetate: hexanes (1:1) to give the
title compound as a yellow solid (0.54 g, 66% yield). ES/MS (m/z) ( Br/ Br):
365.0/367.0 [M+H].
The following compound is prepared essentially by the method of Preparation 26.
Table 9
Preparation 28
(E)-2-[(5-Bromo-3-pyridyl)oxy]-3-(dimethylamino)-l-(4-fluorophenyl)prop-2-en-l-one
Add N,N -dimethylformamide dimethyl acetal (2.8 g, 23 mmol) to a stirred
solution of 2-[(5-bromo-3-pyridyl)oxy]-l-(4-fluorophenyl)ethanone (3.6 g, 12 mmol) in
toluene (50 mL). Stir the mixture under refluxing conditions for 18 hours and evaporate
the solvent under reduced pressure to give the crude title compound as a yellow gummy
solid (4.2 g, 99%), which is used directly without further purification. ES/MS (m/z)
( Br/ Br): 364.9/366.8 [M+H].
The following compound is prepared essentially by the method of Preparation 28.
Table 10
Preparation 30
6-[(E)-2-(Dimethylamino)-l-(4-fluorobenzoyl)vinyloxy]-5-methyl-pyridine-2-carbonitrile
Add N,N -dimethylformamide dimethyl acetal (4.4 g, 37 mmol) to a stirred
solution of 6-[2-(4-fluorophenyl)-2-oxo-ethoxy]-5-methyl-pyridine-2-carbonitrile (1.0 g,
3.7 mmol) in toluene (70 mL). Stir the mixture under refluxing conditions for 40 hours
and evaporate the solvent under reduced pressure to give the crude title compound as a
brown oil (1.2 g, 100%), which is used directly without further purification. ES/MS (m/z)
326 (M+H).
The following compound is prepared essentially by the method of Preparation 30.
Table 11
Preparation 32
3-[[3-(4-Fluorophenyl)-lH-pyrazol-4-yl]oxy]benzonitrile
Add N,N -dimethylformamide dimethyl acetal (1.30 g, 10.9 mmol) to a stirred
solution of 3-[2-(4-fluorophenyl)-2-oxo-ethoxy]benzonitrile (2.55 g, 10.0 mmol) in
toluene (10 mL). Heat the mixture to 120 °C under microwave irradiation with stirring
for 1 hour to give 3-[(E)-2-(dimethylamino)-l-(4-fluorobenzoyl)vinyloxy]benzonitrile:
mass spectrum (m/z): 3 11(M+H), which is directly used without isolation. Add hydrazine
hydrate (0.80 mL, 12.6 mmol) to the reaction and heat the resulting mixture to 120 °C
under microwave irradiation with stirring for 3 hours. Evaporate the solvent and purify
the residue by silica gel flash chromatography, eluting with ethyl acetate: hexanes (1:1) to
give the title compound as a light yellow sticky oil (1.82 g, 65% yield for two steps).
ES/MS (m/z) 280 (M+H).
The following compound is prepared essentially by the method of Preparation 32.
Table 12
Preparation 34
4-Fluoro-3-[[3-(4-fluorophenyl)-lH-pyrazol-4-yl]oxy]benzonitrile
Add N,N -dimethylformamide dimethyl acetal (0.86 g, 7.2 mmol) to a stirred
solution of 4-fluoro-3-[2-(4-fluorophenyl)-2-oxo-ethoxy]benzonitrile (0.79 g, 2.9 mmol)
in toluene (30 mL). Stir the mixture under refluxing condition for 12 hours and evaporate
the solvent under reduced pressure to give a residue. Purify the residue by silica gel flash
chromatography, eluting with ethyl acetate: petroleum ether (1:2) to give 3-[(E)-2-
(dimethylamino)-l-(4-fluorobenzoyl)vinyloxy]-4-fluoro-benzonitrile (0.72 g, 76% yield):
mass spectrum (m/z): 329(M+H). Add hydrazine monohydrochloride (0.26 g, 3.8 mmol)
and triethylamine (0.8 mL, 5.7 mmol) to a stirred solution of 3-[(E)-2-(dimethylamino)-l-
(4-fluorobenzoyl)vinyloxy]-4-fluoro-benzonitrile (0.63 g, 1.9 mmol) in ethanol (20 mL)
and stir the mixture under refluxing condition for 12 hours. Evaporate the solvent and
purify the residue by silica gel flash chromatography, eluting with ethyl acetate:
petroleum ether (1:2) to give the title compound as a white solid (21 mg, 3.7%). ES/MS
(m/z) 298 (M+H).
The following compounds are prepared essentially by the method of Preparation
34.
Table 13
ES/MS
Prep.
Chemical Name Structure (m/z)
No
(M+H)
F
2-Bromo-5-[[3-(4-
( Br/8 1
fluorophenyl) -1HBr)
35 pyrazol-4-
357.9/35 yl]oxy]benzonitrile
9.8
Br
F
3-[[3-(4-Fluorophenyl)-
36 lH-pyrazol-4-yl]oxy]-4- 294
methyl-benzonitrile
F
3-[[3-(4-Fluorophenyl)-
37 lH-pyrazol-4-yl]oxy]-4- 310
methoxy-benzonitrile
H
Preparation 4 1
-(4-Fluorophenyl)-lH-pyrazol-4-yl]oxy]phenyl]methanamine
Add 1M solution of lithium aluminum hydride in THF (1.2 mL, 1.2 mmol)
dropwise to a stirred solution of 3-[[3-(4-fluorophenyl)-lH-pyrazol-4-yl]oxy]benzonitrile
(0.28 g, 0.80 mmol) in THF (5 mL) at 0 °C. Slowly warm to room temperature and stir
for 2 hours. Partition the mixture between ethyl acetate (30 mL) and water (10 mL).
Isolate the organic phase, wash with brine, dry over Na2S0 4, and concentrate under
reduced pressure to give a residue. Purify the residue by silica gel flash chromatography,
eluting with methanol: dicholomethane: ammonium hydroxide (1:9:0.5) to give the title
compound as a white solid (0.12 g, 53%). ES/MS (m/z) 284 (M+H).
Alternate Preparation 4 1
Add Raney nickel (50 mg, 0.57 mmol) to a stirred solution of 3-[[3-(4-
fluorophenyl)-lH-pyrazol-4-yl]oxy]benzonitrile (1.0 g, 3.6 mmol) in 7.0 M ammonium
solution in methanol (60 mL). Degas the reaction vessel three times with hydrogen and
stir the mixture under hydrogen atmosphere for 20 hours. Filter off the solid and
concentrate the filtrate under reduced pressure to give the title compound as a brown oil
(1.0 g, 99%), which is used directly without further purification. ES/MS (m/z) 284
(M+H).
The following compounds are prepared essentially by the method of Alternate
Preparation 41.
Table 14

Methyl 3-
(aminomethyl)-5-[[3-(4-
48 fluorophenyl)-1H- 342
pyrazol-4-
yl]oxy]benzoate
Preparation 49
5-[[3-(4-Fluorophenyl)-lH-pyrazol-4-yl]oxy]-2-(4-methylpiperazin-l-yl)benzonitrile
Under an N2 atmosphere, stir a mixture of N -methylpiperazine (40 mg, 0.40
mmol), 2-bromo-5-[[3-(4-fluorophenyl)-lH-pyrazol-4-yl]oxy]benzonitrile (120 mg, 0.34
mmol), tris(dibenzylideneacetone)dipalladium(0) (37 mmol, 0.04 mmol), 4,5-
bis(diphenylphosphino) -9,9-dimethylxanthene (19 mg, 0.04 mmol), and sodium tbutoxide
(64 mg, 0.67 mmol) in 1,4-dioxane (3 mL) at 100 °C for 20 hours. Filter off the
solid through a pad of diatomaceous earth and evaporate the solvent under reduced
pressure to give a residue. Purify the residue by silica gel flash chromatography, eluting
with methanol: dichloromethane (1:10) to give the title compound as a white solid (90
mg, 71%). ES/MS (m/z) 378 (M+H).
Preparation 50
2-Bromo-6-[[3-(4-fluorophenyl)-1H-pyrazol-4-yl]oxy]pyridine
Add hydrazine monohydrochloride (0. 11 g, 1.6 mmol) to a stirred solution of (E)-
2-[(6-bromo-2-pyridyl)oxy]-3-(dimethylamino)-l-(4-fluorophenyl)prop-2-en-l-one (0.53
g, 1.5 mmol) in ethanol (6 mL). Heat the resulting mixture to 100 °C under microwave
irradiation with stirring for 3 hours and evaporate the solvent under reduced pressure to
give a residue. Purify the residue by silica gel flash chromatography, eluting with a
gradient solvent of 8-25% ethyl acetate in hexanes to give the title compound as a white
solid (0.47 g, 97%). ES/MS (m/z) ( Br/ Br): 334.0/335.9 [M+H].
The following compound is prepared essentially by the method of Preparation 50.
Table 15
Preparation 52
3-Bromo-5-[[3-(4-fluorophenyl)- 1H-pyrazol-4-yl]oxy]pyridine
Add hydrazine hydrate (2.9 g, 58 mmol) to a stirred solution of (E)-2-[(5-bromo-
3-pyridyl)oxy]-3-(dimethylamino)-l-(4-fluorophenyl)prop-2-en-l-one (4.2 g, 12 mmol)
in acetic acid (10 mL). Stir the mixture at room temperature for 2 hours and pour into ice
water (200mL). Extract with ethyl acetate (3 x 50 mL) and wash the combined organic
extracts with brine (2 x 50 mL). Dry over Na2S04 and evaporate the solvent under
reduced pressure to give a residue. Purify the residue by silica gel flash chromatography,
eluting with a gradient solvent of 0-40% ethyl acetate in petroleum ether to give the title
compound as a yellow solid (2.4 g, 62%). ES/MS (m/z) ( Br/ Br): 333.9/335.8 [M+H].
The following compound is prepared essentially by the method of Preparation 52.
Table 16
Preparation 54
6-[[3-(4-Fluorophenyl)-lH-pyrazol-4-yl]oxy]pyridine-2-carbonitrile
Add zinc cyanide (0.22 g, 1.9 mmol), [l,l'-bis(diphenylphosphino)
ferrocene]dichloropalladium(II) (0.11 g, 0.13 mmol), zinc (50 mg, 0.76 mmol) and zinc
acetate (0.13 g, 0.71 mmol) to a stirred solution of 2-bromo-6-[[3-(4-fluorophenyl)-lHpyrazol-
4-yl]oxy]pyridine (0.47 g, 1.4 mmol) in dimethylacetamide (8 mL). Heat the
resulting mixture to 160 °C under microwave irradiation with stirring for 2.5 hours. Filter
off the solid and partition the filtrate between diethyl ether (50 mL) and water (20 mL).
Isolate the organic phase and extract the aqueous phase with diethyl ether (2 x 20 mL).
Combine all the organic extracts, wash with brine, dry over Na2S04 and concentrate
under reduced pressure to give a residue. Purify the residue by silica gel flash
chromatography, eluting with a gradient solvent of 7-35% ethyl acetate in hexanes to
give the title compound as a white solid (54 mg, 14%). ES/MS (m/z) 281 (M+H).
The following compound is prepared essentially by the method of Preparation 54.
Table 17
Preparation 56
5-[[3-(4-Fluorophenyl)-lH-pyrazol-4-yl]oxy]pyridine-3-carbonitrile
Add copper cyanide (0.92 g, 10 mmol) to a stirred solution of 3-bromo-5-[[3-(4-
fluorophenyl)-lH-pyrazol-4-yl]oxy]pyridine (2.3 g, 6.9 mmol) in N -methylpyrrolidone (1
mL). Purge the reaction vessel with nitrogen and heat the mixture to 200 °C under
microwave irradiation with stirring for 30 minutes. Pour the mixture into a stirred
solution of ethylenediamine (40 mL) in water (160 mL) and stir for 15 min. Extract the
mixture with ethyl acetate (3 x 100 mL) and combine all the organic phases. Wash with
brine (2 x 100 mL), dry over Na2S04 and concentrate under reduced pressure to give a
residue. Purify the residue by silica gel flash chromatography, eluting with a gradient
solvent of 0-50% ethyl acetate in petroleum ether to give the title compound as a yellow
solid (1.7 g, 86%). ES/MS (m/z) 281 (M+H).
The following compound is prepared essentially by the method of Preparation 56.
Table 18
Preparation 58
6-[[3-(4-Fluorophenyl)-lH-pyrazol-4-yl]oxy]-5-methyl-pyridine-2-carbonitrile
Add hydrazine hydrate (0.74 g, 15 mmol) to a stirred solution of 6-[(E)-2-
(dimethylamino)-l-(4-fluorobenzoyl)vinyloxy]-5-methyl-pyridine-2-carbonitrile (1.2 g,
3.7 mmol) in acetic acid (10 mL). Stir the mixture at room temperature for 1 hour and
pour into ice water (100 mL). Collect the precipitate by filtration and wash with cold
water (3 x 10 mL). Dissolve the solid in ethyl acetate (50 mL), dry over Na2S04 and
evaporate the solvent under reduced pressure to give the crude title compound as a yellow
solid (0.80 g, 74%), which is used directly without further purification. ES/MS (m/z) 295
(M+H).
The following compounds are prepared essentially by the method of Preparation
58.
Table 19
Preparation 60
[6-[[3-(4-Fluorophenyl)-lH-pyrazol-4-yl]oxy]-2-pyridyl]methanamine
Add Raney nickel (0.3 mL, 3.5 mmol) to a stirred solution of 6-[[3-(4-
fluorophenyl)-lH-pyrazol-4-yl]oxy]pyridine-2-carbonitrile (50 mg, 0.18 mmol) in 7.0 M
ammonium solution in methanol (7 mL). Degas the reaction vessel three times with
hydrogen and stir the mixture under a hydrogen atmosphere for 18 hours. Filter off the
solid through a pad of iato ac o s earth and concentrate the filtrate under reduced
pressure to give the title compound as a brown oil (50 mg, 100%), which is used directly
without further purification. ES/MS (m/z) 285 (M+H).
The following compounds are prepared essentially by the method of Preparation
60.
Table 20
ES/MS Prep.
Chemical Name Structure (m/z)
No
(M+H)
[2-[[3-(4-Fluorophenyl)-
6 1 lH-pyrazol-4-yl]oxy]-4- 285
pyridy1]methanamine
[5-[[3-(4-Fluorophenyl)-
62 lH-pyrazol-4-yl]oxy]-3- 285
pyridy1]methanamine
Preparation 66
Methyl 3-[[3-(4-fluorophenyl)-lH-pyrazol-4-yl]oxy]-5-(ureidomethyl)benzoate
Methyl 3-[[3-(4-fluorophenyl)-lH-pyrazol-4-yl]oxy]-5-(ureidomethyl)benzoate is
prepared substantially as described by Example 1 to give the title compound as a white
solid. ES/MS (m/z) 341 (M+H).
Example 1
-(4-Fluorophenyl)-lH-pyrazol-4-yl]oxy]phenyl]methylurea
Add phenylurethane (70 mg, 0.51 mmol) to a stirred solution of [3-[[3-(4-
fluorophenyl)-lH-pyrazol-4-yl]oxy]phenyl]methanamine (0.12 g , 0.42 mmol) in THF (5
mL) and then add diisopropylethylamine (0.16 mL, 0.92 mmol). Heat the resulting
mixture to 80 °C under microwave irradiation with stirring for 6 hours. Partition between
ethyl acetate (30 mL) and water (10 mL). Isolate the organic phase, wash with brine, dry
over Na2SC>4 and concentrate under reduced pressure to give a residue. Purify the residue
by silica gel flash chromatography, eluting with methanol: dicholomethane (1:9) to give
the title compound as a white solid (0.12 g, 87%). ES/MS (m/z) 327 (M+H).
Alternate Example 1
Add potassium cyanate (0.22 g, 2.7 mmol) to a stirred solution of [3-[[3-(4-
fluorophenyl)- lH-pyrazol-4-yl]oxy]phenyl]methanamine (0.75 g, 2.5 mmol) in a mixture
of methanol (5 mL) and acetic acid (0.1 mL). Heat the resulting mixture to 80 °C under
microwave irradiation with stirring for 1 hour. Evaporate the solvent under reduced
pressure and purify the residue by silica gel flash chromatography, eluting with methanol:
dicholomethane (1:9) to give the title compound as a white solid (0.68 g, 83%). ES/MS
(m/z) 327 (M+H).
The following compounds are prepared essentially by the method of Alternate
Example 1.
Table 2 1
Example 9
-[[3-(4-Fluorophenyl)-lH-pyrazol-4-yl]oxy]-5-(ureidomethyl)benzoic acid
Add lithium hydroxide (0.18 g, 4.4 mmol) to a stirred solution of methyl 3-[[3-(4-
fluorophenyl)-lH-pyrazol-4-yl]oxy]-5-(ureidomethyl)benzoate (0.15 g, 0.44 mmol) in
methanol (15 mL) and water (4 mL). Stir the mixture at room temperature for 20 hours
and evaporate the solvent under reduced pressure to give a residue. Purify the residue by
reverse phase flash chromatography, eluting with acetonitrile and 0.05% NH4HCO 3 in
water in a gradient 0-35% to give the title compound as a white solid (95 mg, 58%).
ES/MS (m/z 371 (M+H).
Example 10
3-[[3-(4-Fluorophenyl)- 1H-pyrazol-4-yl]oxy] -5-(ureidomethyl)benzamide
Add 1-[bis(dimethylamino)methylene] -1H-1,2,3 -triazolo[4,5-bjpyridinium 3-oxid
hexafluorophosphate (67 mg, 0.18 mmol) and diisopropylethylamine (0.12 mL, 0.68
mmol) to a stirred solution of 3-[[3-(4-fluorophenyl)-lH-pyrazol-4-yl]oxy]-5-
(ureidomethyl)benzoic acid (50 mg, 0.14 mmol) and ammonium chloride (36 mg, 0.68
mmol) in DMF (1 mL). Stir the mixture at room temperature for 2 hours. Purify the
mixture by reverse phase flash chromatography, eluting with acetonitrile and 0.05%
NH4HCO 3 in water in a gradient 0-35% to give the title compound as a white solid (35
mg, 70%). ES/MS (m/z) 370 (M+H).
Example 11
[6-[[3-(4-Fluorophenyl)-lH-pyrazol-4-yl]oxy]-2-pyridyl]methylurea
Add potassium cyanate (16 mg, 0.20 mmol) to a stirred solution of [6-[[3-(4-
fluorophenyl)-lH-pyrazol-4-yl]oxy]-2-pyridyl]methanamine (50 mg, 0.17 mmol) in a
mixture of methanol (5 mL) and acetic acid (0.1 mL). Heat the resulting mixture to 80 °C
under microwave irradiation with stirring for 1 hour. Evaporate the solvent under
reduced pressure and purify the residue by silica gel flash chromatography, eluting with a
gradient solvent of 0-10% methanol in dicholomethane to give the title compound as a
white solid (30 mg, 55%). ES/MS (m/z) 328 (M+H).
The following compounds are prepared essentially by the method of Example 11.
Table 22
F
[4-[[3-(4-fluorophenyl)-
14 lH-pyrazol-4-yl]oxy]-2- 328
pyridyljmethylurea
H H NH2
F
[6-[[3-(4-Fluorophenyl)-
lH-pyrazol-4-yl]oxy]-5-
15 342
methyl-2-
pyridyljmethylurea
[6-[[3-(4-Fluorophenyl)-
lH-pyrazol-4-yl]oxy]-3-
16 methyl-2- 342
pyridyljmethylurea
Assays
Enzymatic activity assay of MetAP2 and MetAPl
The compounds exemplified herein are tested essentially as described below and
exhibit an IC50 for the human and mouse MetAP2 assay of lower than 500 nM and are
considered selective for MetAP2 with a MetAPl value greater than 30 .
Full length MetAP2 (human and mouse) and MetAPl (human) proteins are
generated from Sf9 cells using procedure similar to that described in Biochemistry 2003,
42, 5035-5042. MetAP2 and MetAPl are purified in the presence of 5 mM MnCl2 and 2
mM C0Q2 respectively, and stored at -78 °C before use.
Compound inhibition of the catalytic activity of human and mouse MetAP2 in the
present invention is monitored by the formation of the product peptide (Gly-Lys-Val-Lys-
Val-Gly-Val-Asn-Gly) from the substrate peptide (Met-Gly-Lys-Val-Lys-Val-Gly-ValAsn-
Gly) via LC/MS detection. The reaction is typically conducted by incubating the
enzyme, testing compound and substrate (150 ) in a 100 ΐ assay buffer (50 mM
HEPES, 100 mM NaCl, 50 mg/mL BSA, 0.17 mM Triton X-100 at pH 7.5) for 40 min.
After the reaction is stopped by the addition of 200 ΐ acetonitrile, the levels of product
and remaining substrate are quantified on a mass spectrometer. The activity of human
MetAPI is monitored by the formation of the fluorescent product rhodmine-methionine
from the substrate methionine-rhodamine-methionine on a spectrophotometer with the
excitation light at 460 nm and emission light at 535 nm. The reaction is typically
conducted by incubating the enzyme, testing compound and substrate (50 ) in a 100 ΐ
assay buffer (50 mM HEPES, 100 mM NaCl, 0.1% BSA, 0.05% Tween-20, 50 
CoCl2) for 60 min. IC50 value (concentrations of testing compound that provides a50%
inhibition of MetAP2 activity) are calculated typically from a 10-point dose titration
curve using the 4-parameter equation. For Example 1, the IC50 value for hMetAP2 is 11
nM and mMetAP2 is 36 nM. Example 1 has an ICso>30 for MetAPl, demonstrating
selective MetAP2 inhibition as compared with MetAPl. The IC50 for the human and
mouse MetAP2 assay using exemplified compounds is lower than 500 nM and IC50
value for hMetAPl is >30 , demonstrating selective MetAP2 inhibition as compared
with MetAPl.
Therapeutic Weight Loss Effect Measurement of Compounds.
To determine the therapeutic weight loss effects and improvement of metabolic
parameters, compounds from the invention are tested in the high fat diet (HFD) feeding
induced obese mouse model (DIO mice). In this model, C57/B16J male mouse is fed with
the 60% HFD (D12492i, Research Diets) for 16 ~ 28 weeks to establish obesity with
body weight reaching around 50 g. The mice will gradually increase the body weight to
about 50 g and maintain that weight in this obese state. Test compound (via the vehicle
of 0.5% HEC plus 0.25% Tween-80 at 5 mL/kg) is administered orally to the obese DIO
mice once or twice daily throughout the study duration. The dose-dependent weight loss
of obese DIO mice for Example 1 of the oral treatment at 30 mg/kg twice a day is about
6%, 10%, and 14% weight loss compared to the vehicle group at day 7, day 14, and day
2 1 respectively. The data support that the compound of Example 1 is associated with
desired weight loss and offers therapeutic weight loss effect.
The exemplified compounds of the present invention can be readily formulated
into pharmaceutical compositions in accordance with accepted practices known in the art
such as found in Remington's "Pharmaceutical Sciences", Gennaro, Ed., Mack Publishing
Co. Easton Pa. 1990 such as tablets, solid or gel filled capsules, powders, suspensions, or
solutions. The composition can also include one or more pharmaceutically acceptable
carriers, excipients, and diluents.
Preferred pharmaceutical compositions are formulated as a tablet or capsule for
oral administration. The tablet or capsule can include a compound of the present
invention in an amount effective to treat obesity.
The pharmaceutical composition is administered to a patient in amounts effective
to treat obesity. An appropriate amount or dose effective to treat a patient can be
determined by a health care provider.
What is claimed is:
1. A com ound which is
R is selected from the group consisting of H and C¾;
selected from the group consisting of H, CH3, F, CI, OCH 3, C(0)OH,
or a pharmaceutically acceptable salt thereof.
2. A compound or pharmaceutically acceptable salt thereof, as
claimed by Claim 1 wherein X is or
A compound or pharmaceutically acceptable salt thereof, as
claimed by any one of Claims 1 to 2 wherein X is
A compound or salt as claimed by any one of Claims 1 to 3
wherein R is selected from the group consisting of H, F, and CH .
A compound or salt as claimed by any one of Claims 1 to 3
wherein R is
A compound as claimed by any one of Claims 1 to 4 wherein the
compound is [3-[[3-(4-Fluorophenyl)-lH-pyrazol-4-
yl]oxy]phenyl]methylurea, or a pharmaceutically acceptable salt
thereof.
7. A compound or salt as claimed by any one of Claims 1 to 2
wherein X is
8. A compound or salt as claimed by any one of Claims 1, 2, or 7
wherein R is CH
9. A compound as claimed by any one of Claims 1, 2, 7 or 8 wherein
the compound is [6-[[3-(4-Fluorophenyl)-lH-pyrazol-4-yl]oxy]-5-
methyl-2-pyridyl]methylurea or a pharmaceutically acceptable salt
thereof.
10. A method of providing desired weight loss in a mammal in need
thereof, comprising administering an effective amount of a
compound as claimed by any one of Claims 1 to 9, or a
pharmaceutically acceptable salt thereof.
11. A method of treating obesity in a mammal in need thereof,
comprising administering an effective amount of a compound as
claimed by any one of Claims 1 to 9, or a pharmaceutically
acceptable salt thereof.
A method for therapeutic weight loss in a mammal in need thereof,
comprising administering an effective amount of a compound as
claimed by any one of Claims 1 to 9, or a pharmaceutically
acceptable salt thereof.
A compound, or pharmaceutically acceptable salt thereof, as
claimed by any one of Claims 1 to 9 for use in therapy.
A compound, or a pharmaceutically acceptable salt thereof, as
claimed by any one of Claims 1 to 9 for use in the treatment of
obesity.
A compound, or a pharmaceutically acceptable salt thereof, as
claimed by any one of Claims 1 to 9 for use in therapeutic weight
loss.
A pharmaceutical composition comprising a compound as claimed
by any one of Claims 1 to 9 or a pharmaceutically acceptable salt
thereof, and at least one pharmaceutically acceptable carrier,
diluent, or excipient.
20