COMPOUNDS USEFUL FOR INHIBITING CHK1.

The present invention relates to an aminopyrazole compound, or a
pharmaceutically acceptable salt thereof, that inhibits Chkl and is useful for treating
cancers characterized by defects in deoxyribonucleic acid (DNA) replication,
chromosome segregation, and/or cell division.
Chkl is a protein kinase that lies downstream from Atm and/or Atr in the DNA
damage checkpoint signal transduction pathway. In mammalian cells, Chkl is
phosphorylated in response to agents tha cause DNA damage including ionizing
radiation (IR), ultraviolet (UV) light, and hydroxyurea. This phosphorylation which
activates Chkl in mammalian ce ls is dependent on Air. Chkl plays a role in the Atr
dependent DNA damage checkpoint leading to arrest in S phase and at G2M. Chkl
phosphorylates and inactivates Cdc25A, the dual-specificity phosphatase that normally
dephosphorylates cyclin E/Cdk2, halting progression through S-phase. Chkl also
phosphorylates and inactivates Cdc25C, the dual specificity phosphatase that
dephosphorylates cyclin B/Cdc2 (also known as Cdkl) arresting cell cycle progression at
the boundary of G2 and mitosis (Furnari et al., Science, 277:1495-7, 1997). In both cases,
regulation of Cdk activity induces a cell cycle arrest to prevent cells from entering mitosis
in the presence of DNA damage or unreplicated DNA
Various inhibitors of C k l have been reported. In addition, WO 2005/121121
discloses certain aminopyrazole compounds asserted to be modulators of glucose
metabolism.
However, there is still a need for Chk l inhibitors that are potent inhibitors of the
cell cycle checkpoints that can act effectively as potentiators of DNA damaging agents.
The present invention provides compounds that are potent inhibitors of Chk l , which may
be beneficial for the treatment of cancer. The compounds potently abrogate a Chkl
mediated cell cycle arrest induced by treatment with DNA damaging agents in tissue
culture and in vivo. Additionally, the compounds of the present invention provide
inhibition of Chk2, which may be beneficial for the treatment of cancer. Furthermore, the
compounds of the present invention inhibit cell proliferation of cancer cells by a
mechanism dependent on Chkl inhibition. Such new compounds could address the need
for safe and effecti ve treatments of cancer.
T e present invention provides a compound which is (/?)-[5-(2-methoxy-6-
methyi-pyridm-3-yi)-2H-pyrazo or a
pharmaceutically acceptable salt thereof. Preferred embodiments are (i?)-[5-(2-methoxy-
6-methyl-pyridin-3-yl)-2H-pyrazol-3-ylj-[6-(piperidm-3-yloxy)-pyrazi ( ?)-
[5-(2-methoxy-6-methyl-pyridin-3-yl)-2H-pyrazo1-3-yl]-[6-{piperidin-3-yb
2-yl]-amine methane sulfonic acid salt, (i?)-[5-(2-methoxy-6-methyl-pyridin-3-yl)-2Hpyrazol-
3-yl]-[6-(piperidin-3-yloxy)-pyrazin-2-ylj-amine acetic acid salt, (/?)-[5-(2-
methoxy-6-methyl-pyrkim-3-y3)-2H-py
amine hemioxalate salt, and (^)-[5-(2-methoxy-6-methyl-pyridin-3-yl)-2H-pyrazol-3-yl]-
[6-(piperidm-3-yloxy)-pyrazin-2-yi]-amine hemisuccinate salt.
As a particular embodiment, the present invention provides the compound which
is (i?)-[5-(2-methoxy-6-methyl-pyridm-3-y3)-2H-pyrazo3-3-yl]-[6-(piperidin-3-yloxy)-
pyrazin-2-yl]-amine.
The present invention provides the methane sulfonic aci , acetic acid,
hemioxalate, and hemisuccinate salts of (i?)-[5-(2-methoxy-6-methyl-pyridin-3-yl)-2Hpyrazol-
3-yl]-[6-(piperidin-3-yloxy)-pyrazin-2-yl]-amine.
Another embodiment is a hydrate of (ii)~[5-(2-methoxy-6-methyl-pyridin-3-yl)-
2H-pyrazol-3 -yl] -[6-(piperi din-3 -yloxy)-pyrazin-2-yl] -amine.
The present invention provides (/?)-[5-(2-methoxy-6-methyl-pyridin-3-yl)-2Hpyrazol-
3-yl]-[6-(piperidin-3-yloxy)-pyrazin-2-yl]-amine hydrate in crystalline form.
The present invention also provides (i?)-[5-(2~methoxy-6-methyl-pyridin-3-yl)~
2 -pyrazol-3-yl]-[6-(piperidir -3-yloxy)-pyrazin-2-y ]-amine hydrate in crystalline form
characterized by a X-ray powder diffraction pattern having peaks at 2± 0.2 at 5.17 in
combination with one or more of the peaks selected from the group consisting of 5.73,
17.7 1 and 20. 12.
The present invention provides a pharmaceutical composition comprising (R}-[5-
(2-methoxy-6-methyl-pyridin-3 -yl)-2H-pyrazol-3 -yl]-[6-(piperidin-3-yloxy)-pyrazin-2-
yl] -amine, or a pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier, diluent, or excipient.
The present invention provides a pharmaceutical composition comprising (i?)-[5-
(2-methoxy-6-methyl-pyridin-3-yl)-2H-pyrazol-3-yl]-[6-(piperidin-3-yloxy)-pyrazin-2-
yl]-amine, or a p armace ica v acceptable sa t thereof, together with a pharmaceutically
acceptable carrier, diluent, or excipient and optionally other therapeutic ingredients.
The present invention provides a method of treating cancer, comprising
administering to a patient in need thereof an effective amount of (i?)-[5-(2-methoxy-6-
meAyl-pyridm-3-y1)-2H-pyrazol-3-yl]-[6-(piperidin-3-yloxy)-pyrazin-2-yl]-am or a
pharmaceutically acceptable salt thereof. In addition, the present invention also provides
a method of treating cancer, comprising administering to a patient in need thereof an
effective amount of (/?)-[5-(2-meihoxy-6-methyl-pyridin-3-yl)-2H-pyrazol-3-yl]-[6-
(piperidin-3-yloxy)-pyrazin-2-yl]-amine, or a pharmaceutically acceptable salt thereof,
and ionizing radiation. Furthermore, the present invention provides a method of treating
cancer, comprising administering to a patient in need thereof an effective amount of (R)-
[5-(2-methoxy-6-methyl-pyridin-3-yi)-2H-pyrazo1-3-yl]-[6-(pi
2-yl]-amine, or a pharmaceutically acceptable salt thereof and one or more chemotherapy
agents.
The present invention provides the use o i ?)-[5-(2-methoxy-6-methyi-pyridin-3-
yl)-2H-pyrazoI-3-yl]-[6-(piperidin-3-yloxy)-pyrazin-2-yl]-amine, or a pharmaceutically
acceptable salt thereof, for the manufacture of a medicament for the treatment of cancer.
In addition, the present invention also provides the use of (^)-[5-(2-methoxy-6-methylpyridin-
3-yl)-2H-pyrazol-3-yl]-[6-(piperidin-3-yloxy)-pyraz:in-2-yl]-amine, or a
pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the
treatment of cancer wherein said treatment comprises combination therapy with ionizing
radiation. Furthermore, the present invention provides the use of (7?)-[5-(2-methoxy-6-
methyI-pyridm-3-yl)-2H-pyrazQ]-3-yl] or a
pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the
treatment of cancer by combination therapy wherein said combination therapy treatment
comprises administration of said medicament and administration of one or more
chemotherapy agents to the same patient.
The present invention provides (i?)-[5-(2-methoxy-6-methyl-pyridin-3-yl)-2Hpyrazol-
3-y3]-[6-(piperidin-3-yloxy)-pyraz:in-2-yl]-amine, or a pharmaceutically
acceptable salt thereof, for use in therapy. In addition, the present invention also provides
( )-[5-(2-methoxy-6-methyl pyridin 3-yl)-2H-pyrazol-3-yl]-[6-(piperidii -3-yloxy)-
pyrazin-2-yl]-amine, or a pharmaceutically acceptable salt thereof, and ionizing radiation
for use i therapy. Furthermore, the present invention provides (fl)-[5-(2-meihoxy-6-
methyi-pyridm-3-yl)-2H-pyrazol-3-yl]-[6-^ or a
pharmaceutically acceptable salt thereof, and one or more chemotherapy agents for use in
therapy.
The present invention provides (/?)-[5-(2-methoxy-6-methyl-pyridin-3-yl)-2Hpyrazol-
3-yl]-[6-(piperidin-3-yloxy)-pyrazin-2-yl]-amme, or a pharmaceutically
acceptable salt thereof, for use in the treatment of cancer. In addition, the present
invention also provides ( )-[5-(2-met oxy-6-methyl-pyridin-3-yl)-2 -pyrazol-3-yl]-[6-
(piperidm-3-yloxy)-pyrazin-2-yl]-amine, or a pharmaceutically acceptable salt thereof,
and ionizing radiation for use in the treatment of cancer. Furthermore, the present
invention provides (i?)-[5-(2-methoxy-6-methyl-pyridin-3-yl)-2H-pyrazol-3-yl]-[6-
(piperidm-3-yloxy)-pyrazm-2-yl]-amme, or a pharmaceutically acceptable salt thereof
and one or more chemotherapy agents for use in the treatment of cancer.
The present invention provides use of (^)-[5-(2-methoxy-6-methyl-pyridin-3-yl)-
2 -pyrazol-3-yl]-[6-(piperi dir -3-yloxy)-pyrazin-2-yl]-amine, or a pharmaceutically
acceptable salt thereof, for the manufacture of a medicament for the treatment of cancer,
wherein the medicament is to be administered simultaneously, separately, or sequentially
with ionizing radiation.
The present invention provides use of (i?)-[5-(2-meihoxy-6-methyl-pyridin-3-yl)-
2H-pyrazol-3-yl]-[6-(piperidin-3-yloxy)-pyrazin-2-yl]-amine, or a pharmaceutically
acceptable salt thereof, for the manufacture of a medicament for the treatment of cancer,
wherein the medicament also comprises one or more chemotherapy agents or is to be
administered simultaneously, separately, or sequentially with one or more chemotherapy
agents.
The present invention provides (i?)-[5-(2-methoxy-6-methyl-pyridin-3-yl)-2Hpyrazo3-
3-yl]-[6-(piperidin-3-yloxy)-pyrazin-2-yl]-amme, or a pharmaceutically
acceptable salt thereof, for use i simultaneous, separate, or sequential combination with
ionizing radiation in the treatment of cancer.
The present invention provides (i?)-[5-(2-methoxy-6-methyl-pyridin-3-yi)-2Hpyrazol-
3-yl]-[6-(piperidiri-3-yloxy)-pyrazin-2-yl]-amine, or a pharmaceutically
acceptable salt thereof, for use in simultaneous, separate, or sequential combination with
one or more chemotherapy agents in the treatment of cancer.
Furthermore, the present invention provides preferred embodiments of the
methods and uses as described herein, in which the one or more chemotherapy agents is
selected from the group consisting of 5-fiuorouracil, hydroxyurea, gemcitabine,
methotrexate, pemeirexed, doxorubicin, etoposide, cisplatin, and taxol. Additionally, the
present invention provides more preferred embodiments of the methods and uses as
described herein, in which two chemotherapy agents are selected from the group
consisting of 5-fluorouracil, hydroxyurea, gemcitabine, methotrexate, pemeirexed,
doxorubicin, etoposide, cisplatin, and taxol Also, the present invention provides even
more preferred embodiments of the methods and uses as described herein, i which the
chemotherapy agent is selected from the group consisting of 5-fluorouracil, hydroxyurea,
gemcitabine, methotrexate, pemeirexed, doxorubicin, etoposide, cisplatin, and taxol
Preferred embodiments of the methods and uses described herein are cancers selected
from the group consisting of bladder cancer, colon cancer, gastric cancer, liver cancer,
lung cancer, mammary cancer, melanoma, ovarian cancer, pancreatic cancer,
mesothelioma, renal cancer, and uterine cancer.
As used above, and throughout the description of the invention, the following
terms, unless otherwise indicated, shall be understood to have the following meanings:
"Pharmaceutically acceptable salt" or "pharmaceutically acceptable salts" refers to
the relatively non-toxic, inorganic and organic salts of compounds of the present
invention.
The compounds of the present invention are capable of reaction, for example, with
a number of inorganic and organic acids to form pharmaceutically acceptable salts. Such
pharmaceutically acceptable salts and common methodology for preparing t em are well
known in the art. See, e.g., P. Stahl, et aL, Handbook of Pharmaceutical Salts: Properties,
Selection, and Use, (VCHA/Wiiey-VCH, 2002); .M. Berge, ei aL, "Pharmaceutical
Salts", Journal of Pharmaceutical Sciences, Vol. 66, No. 1, January 1977.
The compounds of the present invention are preferably formulated as
pharmaceutical compositions using one or more pharmaceutically acceptable carriers,
diluents, or excipie s and administered by a variety of routes. Preferably, such
compositions are for oral, subcutaneous, or intravenous administration. Such
pharmaceutical compositions and processes for preparing them are well known in the art.
See, e.g.. Remington: The Science and Practice of Pharmacy (A. Gennaro, el aL, eds.,
st ed., Mack Publishing Co., 2005).
The terms "treatment," "treat," "treating," and the like, are meant to include
slowing or reversing the progression of a disorder. These terms also include alleviating,
ameliorating, attenuating, eliminating, or reducing one or more symptoms of a disorder or
condition, even if the disorder or condition is not actually eliminated and even if
progression of the disorder or condition is not itself slowed or reversed.
"Therapeutically effective amount" or "effective amount" means the amount of the
compound, or pharmaceutically acceptable salt thereof, of the present invention or
pharmaceutical composition containing a compound,, or pharmaceutically acceptable salt
thereof, of the present invention that will elicit the biological or medical response of or
desired therapeutic effect on a tissue, system, animal, mammal, or human that is being
sought by the researcher, veterinarian, medical doctor, or other clinician.
The amount of compound of the present invention actually administered will be
determined by a physician under the relevant circumstances, including the condition to be
treated, the chosen route of administration, the actual compound of the present invention
administered, the age, weight, and response of the individual patient, and the severity of
the patient's symptoms. Dosages per day normally fa l within the range of about 0.1 to
about 10 mg kg of body weight. In some instances dosage levels below the lower limit of
the aforesaid range may be more than adequate, while in other cases still larger doses may
be employed.
The compounds of the present invention may be prepared by a variety of
procedures known in the art, as well as those described in the Preparations and Examples
below. The specific synthetic steps for each of the routes described may be combined in
different ways to prepare the compounds of the present invention.
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 an d
heterocyclic chemistry, techniques which are analogous to the syntheses of known
structurally similar compounds and the procedures described in the Preparations and
Examples which follow, including any novel procedures. The following Preparations and
Examples are provided to illustrate the invention in further detail and represent typical
syntheses of the compounds. The names of the compounds of the present invention are
generally provided by ISIS Draw 2.5 SP2 with Autonom add-in.
As used herein, the following terms have the meanings indicated: "BCA" refers to
bicmchoninic acid; "BSA" refers to bovine serum albumin; "DMSO" refers to
dimethylsulfoxide; "DPBS" refers to dibasic phosphate-buffered saline; "DTT" refers to
dithiothreitol; "EtOAc" refers to ethyl acetate;"FBS" refers to fetal bovine serum;
"HEPES" refers to N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid; "MEM" refers
to minimum essential medium; "MeOH" refers to methanol; "PBS" refers to phosphatebuffered
saline; "PI" refers to propid u iodide; "RNAase" refers to ribonuelease A;
"RPMI" refers to Rosweli Park Memorial Institue; "TBST" refers to tris-buffered saline
Tween-20; "THF" refers to tetrahydrofuran; "TR-FRET" refers to time resolved
fluorescent energy transfer; "Tris" refers to tris(hydroxymethyl) ammomethane; "Triton-
X" refers to 4-(l,l,3 3-tetramethyibutyi)phenyi-poiyethylene glycol toctylphenoxypolyethoxyethanol
polyethylene glycol -oeiy pheny ether; and "Tween-
20" refers to poiysorbate 20.
ier/-Butyl (i?)-3-(6-chloropyrazin-2-yl)oxypiperidine-l-carboxylate
Sodium hydride (225.6 g, 5.64 mol) is dispersed into THF (3 L) and the
temperature is lowered to 0 - 5 °C. A solution of (i?)-3-hydroxy-l-boc piperidine (891.6
g, 4.43 mol) in THF (3 L) is added over 1 h while maintaining the temperature between 0
- 5 °C. The reaction is stirred for I h. 2,6-Dichloropyrazine (600 g, 4.03 mol) as a
solution in THF (3 L) is added dropwise over 1.5 h maintaining the same temperature.
The reaction is stirred for 2 h at 25 - 30 °C, and then poured onto ice. The mixture is
diluted with water and extracted with ethyl acetate. The extracts are dried over anhydrous
sodium sulfate, filtered, arsd concentrated. The residual oil is triturated with 5%
dichloromethane in hexane to give the product as a white solid. The solid is collected by
filtration and dried to give 38 g crude material. The crude product is retriturated with
5% dichloromethane in hexanes to give a white solid in quantitative yield. ES/MS m 'z
Preparation 2
2-Methoxy-6-methyl-nicotimc acid methyl ester
To a stirred solution of 2-chloro-6-methyl-nicotinic acid methyl ester ( 10 4 g,
56.52 mmol) in MeOH under nitrogen is added a solution of sodium (2.58 g, 3 04
mmol) in methanol (80.0 mL) (sodium metal is dissolved in methanol under a nitrogen
atmosphere) at room temperature. The reaction mixture is refluxed overnight. The
reaction is cooled to room temperature an the pH is adjusted to pH = 7 with acetic acid
The reaction mixture is diluted with ethyl acetate ( 00 mL) and water (30 mL). The
organic layer is separated and the aqueous layer is extracted with ethyl acetate ( x 75
mL). The combined orgaiiic extracts are dried over a SC>4, filtered, an concentrated to
give crude product. Yield: 7.25 g (71%). MR (400 MHz, CDCI 3) , 8.066 - 8.047
(d, ./ 7.6 Hz, 1H), 6.782 - 6.764 (d, J 7.2 Hz, i . 4.029 is. 3H), 3.879 (s, 3H), 2.483
(s, 3H); ES/MS m/z 82.2 • i .
Preparation 3
5-(2-Methoxy-6-methyl-pyridin-3-yl)-2H-pyrazol-3-ylamine
«-BuLi (1.2 M, 96.0 mL, 5.6 mmol) is added to a solution of acetonitrile (6.08
mL, 5.4 mmol) in THF (300 mL) at - 78 °C and allowed to stir for 30 min at - 78 °C.
2-Methoxy-6-methyl-nicotmic acid methyl ester (20 g, 105.1 mmol) in THF (200 mL) is
added and stirred at - 78 °C for another 30 rrrin. The reaction mixture is quenched at -78
°C with water (500 mL) and washed with EtOAc (2 x 250 mL). The aqueous layer is
separated and evaporated. This is co-distilled twice with toluene to obtain 3-(2-methoxy-
6-methyl-pyridm-3-y1)-3-oxo-propioniirile. Yield = 2 1.4 g (crude). ES/MS m z 191.1
[M+Hf.
A solution of 3-(2-methoxy-6-niethyl-pyridin-3-yl)-3-oxo-propionitrile (21 g,
1 0.4 mmol) in ethanol (200 mL) is placed in a sealed tube. Hydrazine hydrate (32.1 mL,
662.4 mmol) and acetic acid (2 1 0 mL) are added and the reaction heated at 100 C for 2
h. The solvent is evaporated off and the reaction mixture is diluted with EtOAc (500 mL)
and saturated sodium bicarbonate solution (100 mL). The organic layer is separated and
the aqueous layer is extracted with EtOAc (2 x 250 mL). The combined organic extracts
are dried over a2S0 4, filtered, and concentrated to give the crude product which is taken
into the next step without any further purification. Yield 16.5 g (73%). Ή NMR (400
MHz, DMSO- 6 11.50 (bs, ), 7.90 (d, J 7.6 Hz, H), 6.86 (d, J = 7.6 Hz, 1H), 5.88
(s, i f ! . 4.64 (s, 2H), 3.91 (s, 3H), 2.38 (s, 3H).
5-Ainino-3 -(2 -methoxy-6-methyl-pyridin-3-yl)-pyrazole- 1-carboxylic acid r -butyl
ester
A solution of 5-(2-methoxy-6-methyl-pyridin-3-yl)-2H-pyrazol-3-ylamine (16.0 g
78.3 mmol) in THF (200 mL) is added slowly to a stirred suspension of NaH (60% in
mineral oil, 3.4 g, 85.0 mmol) in THF (200 mL) at 0 °C. After 5 min at 0 °C, ά -tertbutyldicarbonate
(19.8 mL, 86 mmol) is added slowly to the reaction mixture and stirred
at 0 °C for 30 mm. The reaction mixture is quenched with ice-water (approximately 250
mL) and the product is extracted into ethyl acetate (2 500 mL) The combined organic
portions are washed with water and saturated NaCl solution (200 mL), dried over
anhydrous a2S0 4, filtered, and concentrated under vacuum to afford crude material.
This material is triturated with hexane twice to obtain 8.5 g (78%) of the title compound.
NMR (400 MHz, DMSO- d6 8.05 (d, J = 7.6 Hz, . 6.90 id. ./ 7.2 Hz, i f •. 6.28
(s, 2H), 5 85 (s, 1H), 3 90 (s, 3H), 2 40 (s, 3H), 1.56 (s, 9H).
Preparation 5
( ?)-3- {6-[2- r t-Butoxycarbonyl-5-(2-memoxy-6-
ylamino]-pyrazin-2-yloxy}-piperidme-l-carboxylic acid r t-butyl ester
A mixture of 5-ammo-3-(2-methoxy-6-metliy3-pyridin-3-yl)-pyrazole-l -
carboxylic acid tert-bxityl ester (50.0 g, 164.5 mmol). tert-b y (J¾)-3-(6-chIoropyrazin-2-
yl)oxypiperidine- -carboxylate (56.6 g, 180.9 mmol), 4,5-bis-diphenylphosphanyl-9,9-
dimethyl-9H-xantbene (14.2 g, 24.6 mmol) and Cs2C0 3 (85.5 g, 263 mmol) in 1,4-
dioxane (1.4 L) is equally divided into two side by side round bottom flasks and both are
purged with argon for 2 h . Pd(OAc) 2 (5.4 g 24.6 mmol) is added (half to each vessel)
and purging continues for 1 h. The reactions are then heated at 90-95 C for 1 h. The
reaction mixtures are cooled to room temperature, combined, and diluted with ethyl
acetate ( 1 L). The mixture is then filtered through diatomaeeous earth, washed with ethyl
acetate and the filtrate is concentrated. The crude product is purified on silica gel with
15% EtOAc/hexane as eluent to provide 55 g (57% yield) of a white powder. The 55 g of
purified product is combined with 15 g of similarly prepared and purified material
(obtained from 20 g of 5-ammo-3-(2-methoxy-6-methyl~pyridin-3-yl)-pyrazole-l -
carboxylic acid ter/-buty l ester). The combined 70 g of material is dissolved in a 4:1
mixture of THF and methanol ( 1.4 L) and treated with QuadraSil™ AP ( 140 g) for 2 h.
The reaction mixture is filtered through diatomaeeous earth and washed with ethyl acetate
(4 00 mL). The filtrate is again stirred with QuadraSil™ AP (140 g) for h and
fi ltered as above. The solvent is evaporated to give the title compound as a white solid.
Yield = 70 g (47%). ES/MS m/z 582.5 [M+H]+.
Example 1
(/?)-r5~(2-Methoxy~6~methy]-pyri
razin-2-y ] -amine
To a stirred solution of (i?)-3-{6-[2-^ri-butoxycarbonyl-5-(2~methoxy-6-methylpyridin-
3-yl)-2H-pyrazol-3-ylamino]-pyrazin-2-yloxy}-piperidine-l-carboxylic acid iertbutyl
ester ( 3.0 g, 22.3 mmol) in dichloromethane (150 ml.) is added a solution of
trifluoroaeetic acid (12.4 mL, 167 mmol) in dichloromethane (20 mL) over a period of 5
mi at 0 °C. The reaction is allowed to warm to room temperature and stirred for 3 h.
The reaction is diluted with dichloromethane (1000 mL), followed by addition of
saturated sodium bicarbonate solution (250 mL) and then stirred for 4 h. The organic
portion is separated an d dried over anhydrous sodium sulfate, filtered, and evaporated.
The resulting material is ciystallized from isopropanol to obtain the desired product.
Yield = 7.2 g (85%). ί MR (400 MHz, DMSO- ) 12.40 (s, H ). 9.71 (s, H ). 8.02
(d, 7.6 Hz, I}. 7 97 (s, . 7 46 (s, 1H), 6.93 (d, 7.6 Hz, I). 6.91 (s, I). 4 94-
4.86 (m, 1H), 3.97 (s, 3H), 3.20-3.13 (m, 1H), 2.83-2.75 (m, 1H), 2.57 (dd, j 2.0. 8.4
Hz, IH), 2.53-2.45 (m obscured, 1H), 2.42 (s, 3H), 2.15-2.05 (m, 1H), 1.71-1.63 (m, 1H),
1.60-1 .49 (m, HI). 1.49-1.40 (m,lH); ES/MS m/z 382.5 [V! • IS .
(i?)-[5-(2-Methoxy-6-methyl-pyridin-3-yl)-2H-pyrazol-3-y3]-[6-(piperidin-3-yloxy)-
pyrazin--2--yl] -amine methane sulfonic acid salt
Methane sulfonic acid (0.247 g, 2.57 mmol) is added to a stirred solution of (i?)-
[5-(2-methoxy-6-methyl-pyridm-3-yl)-2H-py^
2-yl]-amine (0.982 g, 2.57 mmol) in dichloromethane (25 mL) at 0 °C, The reaction is
aliowed to warm to room temperature and agitated for 45 min. The solvent is evaporated,
and the resulting salt is washed with ether ( 0 mL) and peniane ( 10 mL) sequentially to
obtain the desired product. Yield 1.13 9 g (92.6%). Ή NMR (400 MHz, DM80- TicUn~3-yl)-2H^yrazol-3-yl]-[6-(piperidin-3-yloxy)-pyrazin-2~yl]-amine.
3. The compound according to Claim 1 which is (i?)-[5-(2-methoxy-6-
methyl-pyridin-3-yl)-2H-pyrazol-3-yl]-[6-{piperidm-3-yloxy)-pyrazin-2-yl]-a^
methane sulfonic acid salt.
4. The compound according to Claim 1 which is (ii)-[5-(2-methoxy-6-
methyl-pyridin-3-yl)-2H-pyrazol-3-yl]-[6-(piperidm-3-yloxy)-pyrazin-2-yl]-am acetic
acid salt.
5. The compound according to Claim 1 which is (i?)-[5-(2-methoxy-6-
methyl-pyridin-3-yl)-2H-pyTazol-3-yl]-[6-(piperidin-3-yloxy)-pyrazin-2-yl]-amine
hemioxalate sail
6. The compound according to Claim 1 which is (i?)-[5-(2-meihoxy-6-
methyi-pyridin-3-yl)-2H-pyrazoI-3-yl]-[6-(p^
hemisuccinaie salt.
7. A pharmaceutical composition comprising the compoursd or salt according
to any one of Claims 1-6, and a pharmaceutically acceptable carrier, diluent, or excipient.
8. A method of treating cancer, comprising administering to a patient in need
thereof an effective amount of the compound or salt according to any one of Claims 1-6.
9. A method of treating cancer, comprising administering to a patient in need
thereof an effective amount of the compound or salt according to any one of Claims 1-6
and ionizing radiation.
10. A method of treating cancer, comprising administering to a patient in need
thereof an effective amount of the compound or salt according to any one of Claims 1-6
and one or more chemotherapy agents.
. The method according to Claim 0, wherein the one or more
chemotherapy agents is selected from the group consisting of 5-fluorouracil, hydroxyurea,
gemcitabine, methotrexate, pemetrexed, doxorubicin, etoposide, cisplatin, and taxol.
2. The method according to any one of Claims 8- 1, wherein the cancer is
selected from the group consisting of bladder cancer, colon cancer, gastric cancer, liver
cancer lung cancer, mammary cancer melanoma, ovarian cancer, pancreatic cancer,
mesothelioma, renal cancer, and uterine cancer.
13. The compound or salt according to any one of Claims 1-6 for use in
therapy.
14. The compound or salt according to any one of Claims 1-6 for use in the
treatment of cancer.
5. The compound or salt according to any one of Claims 1-6 for use in
simultaneous, separate or sequential combination with ionizing radiation in the treatment
of cancer.
16. The compound or salt according to any one of Claims 1-6 for use in
simultaneous, separate, or sequential combination with one or more chemotherapy agents
in the treatment of cancer.
17. The compound or salt for use according to Claim 16, wherein the one or
more chemotherapy agents is selected from the group consisting of 5-fluorouracil,
hydroxyurea, gemcitabine, methotrexate, pemetrexed, doxorubicin, etoposide. cisplatm,
and taxol.
18. The compound or salt for use according to any one of Claims 14-17,
wherein the cancer is selected from the group consisting of bladder cancer, colon cancer,
gastric cancer, liver cancer, lung cancer, mammary cancer, melanoma, ovarian cancer,
pancreatic cancer, mesothelioma, renal cancer, and uterine cancer.
19. A pharmaceutical composition comprising the compound or salt according
to any one of Claims 1-6, together with a pharmaceutically acceptable carrier, diluent, or
excipient and optionally other therapeutic ingredients.