U.S. patent application number 14/672477 was filed with the patent office on 2015-09-24 for pyrazolylaminopyridines as inhibitors of fak.
The applicant listed for this patent is GlaxoSmithKline LLC. Invention is credited to Jerry Leroy ADAMS, Thomas H. Faitg, Neil W. Johnson, Hong Lin, Xin Peng.
Application Number | 20150265589 14/672477 |
Document ID | / |
Family ID | 42132177 |
Filed Date | 2015-09-24 |
United States Patent
Application |
20150265589 |
Kind Code |
A1 |
ADAMS; Jerry Leroy ; et
al. |
September 24, 2015 |
PYRAZOLYLAMINOPYRIDINES AS INHIBITORS OF FAK
Abstract
The present invention relates to a compound of formula (I):
##STR00001## or a pharmaceutically acceptable salt thereof, wherein
R.sup.1, R.sup.2, R.sup.3, R.sup.11, R.sup.12, R.sup.13, Q, Z, and
p are as described herein. Compounds of the present invention are
useful for the treatment of cancers.
Inventors: |
ADAMS; Jerry Leroy;
(Collegeville, PA) ; Faitg; Thomas H.;
(Collegeville, PA) ; Johnson; Neil W.;
(Collegeville, PA) ; Lin; Hong; (Collegeville,
PA) ; Peng; Xin; (Collegeville, PA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
GlaxoSmithKline LLC |
Wilmington |
DE |
US |
|
|
Family ID: |
42132177 |
Appl. No.: |
14/672477 |
Filed: |
March 30, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14047688 |
Oct 7, 2013 |
9012479 |
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14672477 |
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13126064 |
Apr 26, 2011 |
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PCT/US09/62163 |
Oct 27, 2009 |
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14047688 |
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61242423 |
Sep 15, 2009 |
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61178517 |
May 15, 2009 |
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61108568 |
Oct 27, 2008 |
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Current U.S.
Class: |
514/341 |
Current CPC
Class: |
A61K 31/4439 20130101;
A61P 35/00 20180101; A61P 43/00 20180101; A61K 31/337 20130101;
G01N 33/57423 20130101; A61K 31/4375 20130101; C07D 401/12
20130101; A61K 31/47 20130101; C07D 401/14 20130101 |
International
Class: |
A61K 31/4439 20060101
A61K031/4439 |
Claims
[0338] 1. The method of inhibiting FAK activity in a mammal in need
thereof, which comprises administering to such mammal a
therapeutically effective amount of the compound:
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-N-(methyloxy)benzamide; or a pharmaceutically
acceptable salt thereof.
2. The method of claim 1 wherein the mammal is a human.
3. A method of treating or lessening the severity of cancer in a
mammal in need thereof, which comprises administering to such
mammal a therapeutically effective amount of the compound:
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-N-(methyloxy)benzamide; or a pharmaceutically
acceptable salt thereof.
4. A method according to claim 3 wherein said cancer is selected
from: skin, breast, brain, cervical carcinomas, testicular
carcinomas, astrocytic, colorectal, endometrial, esophageal,
gastric, head and neck, hepatocellular, laryngeal, lung, oral,
ovarian, prostate, thyroid carcinomas, sarcomas, lung, lymphomas,
mesothelioma, Gastrointestinal, stomach, pancreas, small bowel,
large bowel, kidney, bladder and urethra, liver, bone, Ewing's
sarcoma, malignant lymphoma, multiple myeloma, malignant giant cell
tumor chordoma, brain, glioblastoma multiform, spinal cord, uterus,
cervical, ovarian, squamous cell carcinoma, and hematologic
cancers.
5. A method according to claim 4 wherein said cancer is brain
cancer.
6. A method according to claim 4 wherein said cancer is
mesothelioma.
7. A method according to claim 4 wherein said cancer is pancreatic
cancer.
Description
[0001] This application is a Continuation of U.S. application Ser.
No. 14/047,688 filed 7 Oct. 2013, which is a continuation of U.S.
application Ser. No. 13/126,064 filed 26 Apr. 2011, which is a 371
of International Application No. PCT/US09/62163 filed 27 Oct. 2009,
which claims priority from U.S. Provisional Application No.
61/242,423 filed 15 Sep. 2009, which claims priority from U.S.
Provisional Application No. 61/178,517 filed 15 May 2009; which
claims priority from U.S. Provisional Application No. 61/108,568
filed 27 Oct. 2008, which are incorporated herein in their
entirety.
AREA OF THE INVENTION
[0002] This invention relates to a class of pyrazolylaminopyridines
that inhibit Focal Adhesion Kinase (FAK), as well as compositions
thereof. Compounds of the present invention are useful in the
treatment of proliferative diseases including, but not limited to
cancer.
BACKGROUND OF THE INVENTION
[0003] Tyrosine kinases play an important role in the regulation of
many cell processes including cell proliferation, cell survival,
and cell migration. It is known that certain tyrosine kinases
become activated by mutation or are abnormally expressed in many
human cancers. For example, the epidermal growth factor receptor
(EGFR) is found mutated and/or overexpressed in breast, lung,
brain, squamous cell, gastric, and other human cancers. Selective
inhibitors of the tyrosine kinase activity of EGFR have been shown
to be of clinical value in treatment of cancers with mutated and/or
overexpressed EGFR. Thus, selective inhibitors of particular
tyrosine kinases are useful in the treatment of proliferative
diseases such as cancer.
[0004] FAK (encoded by the gene PTK2) is a non-receptor tyrosine
kinase that integrates signals from integrins and growth factor
receptors. FAK has been reported to play a role in the regulation
of cell survival, growth, adhesion, migration, and invasion (McLean
et al 2005, Nat Rev Cancer 5:505-515). Furthermore, FAK is
regulated and activated by phosphorylation on multiple tyrosine
residues. Overexpression of FAK mRNA and/or protein has been
documented in many solid human tumors, including but not limited
to, cancers of the breast, colon, thyroid, lung, ovary, and
prostate; but also including cancers of hematological origin,
including but not limited to leukemia such as acute myeloid
leukemia (AML). (Owens et al. 1995, Cancer Research 55: 2752-2755;
Agochiya et al. 1999, Oncogene 18: 5646-5653; Gabarro-Niecko et al.
2003, Cancer Metastasis Rev. 22:359-374; Recher et al. 2004, Cancer
Research 64:3191-3197; Zhao and Guan, 28:35-49, 2009, Cancer
Metastasis Rev.). More significantly, there is evidence that
phosphorylated FAK is increased in malignant compared to normal
tissues (Grisaru-Granovsky et al. 2005, Int. J. Cancer 113:
372-378) and could represent a prognostic marker of metastasis. FAK
activity is clearly implicated in advanced and metastatic human
cancer (Zhao and Guan, 28:35-49, 2009, Cancer Metastasis Rev.).
[0005] Elimination of FAK by RNAi or expression of a FAK dominant
negative has been shown to induce loss of adhesion and cell death
in human breast and melanoma cell lines, and to augment
docetaxel-mediated apoptosis in ovarian cancer cells (Beviglia et
al 2003, Biochem J. 373:201-210, Smith et al 2005, Melanoma Res.
15:357-362, Halder et al 2005, Clin. Cancer Res. 11:8829-8836).
However, inhibition of FAK in normal human fibroblasts or
immortalized mammary cells (MCF 10A) was found not to cause loss of
attachment or apoptosis (Xu et al. 1996 Cell Growth and Diff
7:413-418). Inhibition of FAK by dominant negative expression has
also been shown to reduce tumor growth and eliminate lung
metastasis of mammary adenocarcinoma cells in a syngeneic rat model
(van Nimwegen et al 2005, Cancer Res. 65:4698-4706). Similarly,
inhibition of FAK by shRNA inhibited lung metastasis and reduced
lethality by 40% in a syngeneic mouse model (Mitra et al 2006,
Oncogene 25: 4429-4440). In this study, transient re-expression of
wild-type, but not kinase-dead FAK, reversed the shRNA phenotypes
Inhibition of FAK by dominant negative expression in mouse 4T 1
carcinoma cells reduced tumor growth and angiogenesis in mice
(Mitra et al 2006, Oncogene 25:5969-5984). Furthermore, loss of FAK
catalytic activity (reconstitution of FAK-/- cells with kinase-dead
FAK) reduced growth of v-Src tumors in mice and decreased
angiogenesis.
[0006] Thus, there is strong evidence to suggest that inhibition of
FAK activity induces apoptosis, loss of adhesion, inhibition of
cell growth and migration, and that such inhibition reduces
angiogenesis. Accordingly, compounds that inhibit FAK activity
would be useful for the treatment of cancer.
SUMMARY OF THE INVENTION
[0007] The present invention relates to compounds of formula
(I):
##STR00002##
[0008] or a salt thereof, wherein:
[0009] R.sup.1 is halo, CF.sub.3, C.sub.1-C.sub.6-alkyl,
isopropenyl, (C.sub.2-C.sub.6-alkylene) C.sub.3-C.sub.6-cycloalkyl,
C.sub.1-C.sub.6-alkoxy, or cyano;
[0010] in R.sup.2 when p is other than 0, each R.sup.2 is
independently F, Cl, CF.sub.3, methyl, methoxy, CH.sub.2CF.sub.3,
--(X).sub.q--C.sub.1-C.sub.4-alkylene-R.sup.4,
--(X--C.sub.1-C.sub.4-alkylene).sub.q-NR.sup.5--C(O)--R.sup.6,
--(X--C.sub.1-C.sub.4-alkylene).sub.q-(NR.sup.5).sub.q--SO.sub.x--R.sup.7-
, --(X--C.sub.1-C.sub.4-alkylene).sub.q-Y--N(R.sup.8).sub.2; a 5-
to 6-membered heterocycloalkyl-(R.sup.9).sub.q group, or a 5- to
6-membered)heteroaryl-(R.sup.10).sub.r group;
[0011] R.sup.3 is independently H, C.sub.3-C.sub.6-cycloalkyl,
C.sub.1-C.sub.6-alkyl, C.sub.1-C.sub.6alkoxy,
C.sub.1-C.sub.6-alkylene-R.sup.4,
O--C.sub.1-C.sub.6-alkylene-R.sup.4, or, the R.sup.3 groups,
together with Z, form a 5- to 6-membered cyclic ring optionally
substituted with methyl, C.sub.1-C.sub.4-alkylene-R.sup.4, or
C.sub.3-C.sub.6-cycloalkyl;
[0012] R.sup.4 is H, -(Q).sub.q-N(R.sup.8).sub.2, OH, SH,
C.sub.1-C.sub.6-alkoxy, C.sub.1-C.sub.6-thioalkyl, or a 5- to
6-membered heterocycloalkyl-(R.sup.9).sub.q group;
[0013] R.sup.5 is H or C.sub.1-C.sub.6-alkyl;
[0014] R.sup.6 is H, C.sub.1-C.sub.6-alkyl, C.sub.1-C.sub.6-alkoxy,
N(R.sup.8).sub.2, or a 5- to 6-membered)
heteroaryl-(R.sup.10).sub.r group;
[0015] R.sup.7 is C.sub.1-C.sub.6-alkyl, phenyl-(R.sup.9).sub.q, or
5- to 6-membered)heteroaryl-(R.sup.10).sub.r
[0016] R.sup.8 is independently H, C.sub.1-C.sub.6-alkyl,
--O--C.sub.1-C.sub.6-alkyl or, together with the nitrogen atom to
which they are attached, form a 5- or 6-membered heterocycloalkyl
group;
[0017] R.sup.9 is H, C.sub.1-C.sub.6-alkyl, C.sub.1-C.sub.6-alkoxy,
-(Q).sub.q-N(R.sup.8).sub.2, -Q-C.sub.1-C.sub.6-alkyl,
--C.sub.1-C.sub.6alkylR.sup.4, or 5- to 6-membered
heterocycloalkyl;
[0018] R.sup.10 is H, C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxy, or -Q-C.sub.2-C.sub.6-alkyl;
[0019] R.sup.11 is C.sub.1-C.sub.6-alkyl, CF.sub.3,
--CH.sub.2CF.sub.3, -(Q).sub.q-C.sub.1-C.sub.4-alkylene-R.sup.4,
-Q-N(R.sup.8).sub.2, phenyl-(R.sup.5).sub.s, a 5- to 6-membered
heterocycloalkyl-(R.sup.9).sub.q group, or a 5- to 6-membered
heteroaryl-(R.sup.10).sub.r group;
[0020] R.sup.12 is H, C.sub.1-C.sub.6-alkyl, F, Cl, CF.sub.3, OH,
CN, nitro, COOH, --COO--C.sub.1-C.sub.6-alkyl,
--Y--N(R.sup.8).sub.2, C.sub.3-C.sub.6-cycloalkyl-R.sup.14,
--(X).sub.q--C.sub.1-C.sub.6-alkylene-R.sup.4,
--(X--C.sub.1-C.sub.6-alkylene).sub.q-NR.sup.5--C(O)--R.sup.6,
--(X--C.sub.1-C.sub.6-alkylene).sub.q-(NR.sup.5).sub.q--SO.sub.x--R.sup.7-
, --(X--C.sub.1-C.sub.6-alkylene).sub.q-Y--N(R.sup.8).sub.2,
heterocycloalkyl-(R.sup.9).sub.q, heteroaryl-(R.sup.10).sub.r, or
phenyl-(R.sup.15).sub.s;
[0021] R.sup.13 is H, F, Cl, C.sub.1-C.sub.6-alkyl, or
C.sub.3-C.sub.6-cycloalkyl; or R.sup.12 and R.sup.13, together with
the carbon atoms to which they are attached, form a fused 5- or
6-membered carbocycloalkyl or heterocycloralkyl group;
[0022] R.sup.14 is independently H, C.sub.1-C.sub.6-alkyl.
--NR.sup.5--SO.sub.2--R.sup.7, --Y--N(R.sup.8).sub.2, or
--(X).sub.q--C.sub.1-C.sub.6-alkylene-R.sup.4;
[0023] R.sup.15 is independently F, Cl, CF.sub.3,
C.sub.1-C.sub.3-alkyl, or C.sub.1-C.sub.3-alkoxy;
[0024] p is 0, 1, 2, or 3; [0025] q is 0 or 1; [0026] r is 0, 1, or
2; [0027] s is 0, 1, 2, or 3; [0028] x is 1 or 2;
[0029] Q is --C(O)--, --S(O)--, or --SO.sub.2--;
[0030] X is NR.sup.5, O, S, --S(O)--, or --SO.sub.2--;
[0031] Y is a bond, SO.sub.2, or C(O); and
[0032] Z is N or CR.sup.5.
[0033] In a further embodiment, the present invention relates to a
composition comprising a) the compound of formula (I) or a
pharmaceutically acceptable salt thereof; and b) a pharmaceutically
acceptable excipient.
[0034] In a further embodiment, the present invention relates to a
method of treating a proliferative disease such as cancer or an
abnormal angeogensis disease such as macular degeneration,
comprising administering to a patient in need thereof a
pharmaceutically effective amount of the compound of formula
(I).
DETAILED DESCRIPTION OF THE INVENTION
[0035] In another aspect, the present invention relates to
compounds of formula (Ia):
##STR00003##
[0036] wherein the various groups are the same as set out above for
formula (I); or a pharmaceutically acceptable salt thereof.
[0037] In another aspect, the present invention is represented by a
compound of formula (Ib):
##STR00004##
[0038] wherein the various groups are the same as set out above for
formula (I); or a pharmaceutically acceptable salt thereof.
[0039] In another aspect of the present invention, Q is C(O) and Z
is N.
[0040] In another aspect of the present invention, R.sup.1 is Cl,
CF.sub.3, or CN;
[0041] In another aspect of the present invention, R.sup.2 is
F;
[0042] In another aspect of the present invention, one R.sup.3 is
methyl and the other R.sup.3 is H;
[0043] In another aspect of the present invention, one R.sup.3 is
methoxy and the other R.sup.3 is H;
[0044] In another aspect of the present invention, R.sup.1 is
C.sub.1-C.sub.6-alkyl;
[0045] In another aspect of the present invention, R.sup.12 is
C.sub.1-C.sub.6-alkyl, hydroxymethyl, or cyclopropyl;
[0046] In another aspect of the present invention, R.sup.13 is
H;
[0047] In another aspect of the present invention, p is 0 or 1;
[0048] As used herein, "halo" refers to fluoro, chloro, or
bromo.
[0049] "C.sub.1-C.sub.6-alkyl" refers to a linear or branched alkyl
group including methyl, ethyl, n-propyl, isopropyl, n-butyl,
isobutyl, t-butyl, n-pentyl, and n-hexyl.
[0050] "C.sub.1-C.sub.6-alkoxy" refers to C.sub.1-C.sub.6-alkyl-O--
groups, including methoxy, ethoxy, n-propoxy, iso-propoxy, and
n-butoxy groups.
[0051] The term "alkylene" (e.g., C.sub.2-C.sub.4-alkylene or
C.sub.1-C.sub.6-alkylene) refers to a linear or branched
hydrocarbon radical having the specified number of carbon atoms.
The group "-alkylene-R.sup.4" refers to a substituted or
unsubstituted alkyl group having the specified number of carbon
atoms; thus, where R.sup.4 is H, "alkylene" is synonymous with
"alkyl"; otherwise, alkylene is a bivalent radical. Examples of
--(X).sub.q--C.sub.2-C.sub.4-alkylene-R.sup.4 include
--CH.sub.2CH.sub.2--N(CH.sub.3).sub.2, --CH.sub.2CH.sub.2--OH,
--CH.sub.2CH(CH.sub.3)--OCH.sub.3,
--N(CH.sub.3)--CH.sub.2CH.sub.2CH.sub.2-piperidinyl;
--O--CH.sub.2CH(CH.sub.3)--OCH.sub.3; and the like.
[0052] C.sub.3-C.sub.6-cycloalkyl refers to a cyclopropyl,
cyclobutyl, cyclopentyl, or cyclohexyl group.
[0053] As used herein, "5- or 6-membered heterocycloalkyl" refers
to a 5- or 6-membered cycloaliphatic group that includes an O, N,
or S heteroatom or a combination thereof. Examples of suitable
heterocycloalkyl groups include pyrrolidinyl, pyrrolidinonyl,
piperidinyl, piperazinyl, oxopiperazinyl, morpholino, and
thiomorpholino groups.
[0054] The R.sup.8 groups may, together with the nitrogen atom to
which they are attached, form a 5- to 6-membered cyclic ring,
examples of which include pyrrolidinyl, pyrrolidinonyl,
piperidinyl, piperazinyl, oxopiperazinyl, morpholino, and
thiomorpholino groups.
[0055] The term "heteroaryl" refers to a 5- or 6-membered aromatic
group containing at least one N, O, or S atom. Examples of suitable
heteroaryl groups include pyridinyl, pyridazinyl, pyrimidinyl,
pyrazinyl, pyrrolyl, furyl, thienyl, pyrazolyl, imidazolyl,
furazanyl, oxazolyl, thiazolyl, isoxazolyl, 1,2,3-triazolyl,
1,2,4-triazolyl, tetrazolyl, and isothiazolyl.
[0056] As used herein, "pharmaceutically acceptable" refers to
those compounds, materials, compositions, and dosage forms which
are, within the scope of sound medical judgment, suitable for use
in contact with the tissues of human beings and animals without
excessive toxicity, irritation, or other problem or
complication.
[0057] The skilled artisan will appreciate that pharmaceutically
acceptable salts of compounds of formula (I) may be prepared. More
particularly, inasmuch as compounds according to formula (I)
contain a basic functional group--and may include an acid
functional group--they are capable of forming pharmaceutically
acceptable salts by treatment with a suitable acid or base.
Suitable acids include pharmaceutically acceptable inorganic acids
and organic acids. Representative pharmaceutically acceptable acids
include hydrogen chloride, hydrogen bromide, nitric acid, sulfuric
acid, sulfonic acid, phosphoric acid, acetic acid, hydroxyacetic
acid, phenylacetic acid, propionic acid, butyric acid, valeric
acid, maleic acid, acrylic acid, fumaric acid, malic acid, malonic
acid, tartaric acid, citric acid, salicylic acid, benzoic acid,
tannic acid, formic acid, stearic acid, lactic acid, ascorbic acid,
p-toluenesulfonic acid, oleic acid, and lauric acid.
[0058] Suitable bases include inorganic bases, such as hydrides,
hydroxides and carbonates of lithium, sodium, potassium, calcium,
magnesium, and zinc, as well as organic bases such as arginine,
choline, diethylenetriamine, dimethylamine, ethylenediamine,
imidazole, lysine, morpholine, proline, and trimethylamine.
[0059] As used herein, the term "a compound of formula (I)" or "the
compound of formula (I)" refers to one or more compounds according
to formula (I). The compound of formula (I) may exist in a
crystalline or noncrystalline form, or as a mixture thereof. The
skilled artisan will appreciate that pharmaceutically acceptable
solvates may be formed for crystalline compounds wherein solvent
molecules are incorporated into the crystalline lattice during
crystallization. The incorporated solvent molecules may be water
molecules or non-aqueous such as ethanol, isopropanol, DMSO, acetic
acid, ethanolamine, and ethyl acetate molecules. Crystalline
lattice incorporated with water molecules are typically referred to
as "hydrates." Hydrates include stoichiometric hydrates as well as
compositions containing variable amounts of water. The present
invention includes all such solvates.
[0060] Certain of the compounds described herein may contain one or
more chiral atoms, or may otherwise be capable of existing as two
enantiomers. The compounds claimed below include mixtures of
enantiomers as well as purified enantiomers or enantiomerically
enriched mixtures. Also included within the scope of the invention
are the individual isomers of the compounds represented by formula
(I), or claimed below, as well as any wholly or partially
equilibrated mixtures thereof. The present invention also covers
the individual isomers of the claimed compounds as mixtures with
isomers thereof in which one or more chiral centers are
inverted.
[0061] Where there are different isomeric forms they may be
separated or resolved one from the other by conventional methods,
or any given isomer may be obtained by conventional synthetic
methods or by stereospecific or asymmetric syntheses.
[0062] While it is possible that, for use in therapy, a compound of
formula (I), as well as salts, solvates and the like, may be
administered as a neat preparation, i.e. no additional carrier, the
more usual practice is to present the active ingredient confected
with a carrier or diluent. Accordingly, the invention further
provides pharmaceutical compositions, which includes a compound of
formula (I) and salts, solvates and the like, and one or more
pharmaceutically acceptable carriers, diluents, or excipients. The
compounds of formula (I) and salts, solvates, etc, are as described
above. The carrier(s), diluent(s) or excipient(s) must be
acceptable in the sense of being compatible with the other
ingredients of the formulation and not deleterious to the recipient
thereof. In accordance with another aspect of the invention there
is also provided a process for the preparation of a pharmaceutical
formulation including admixing a compound of the formula (I), or
salts, solvates etc, with one or more pharmaceutically acceptable
carriers, diluents or excipients.
[0063] It will be appreciated by those skilled in the art that
certain protected derivatives of compounds of formula (I), which
may be made prior to a final deprotection stage, may not possess
pharmacological activity as such, but may, in certain instances, be
administered orally or parenterally and thereafter metabolised in
the body to form compounds of the invention which are
pharmacologically active. Such derivatives may therefore be
described as "prodrugs". Further, certain compounds of the
invention may act as prodrugs of other compounds of the invention.
All protected derivatives and prodrugs of compounds of the
invention are included within the scope of the invention. It will
further be appreciated by those skilled in the art, that certain
moieties, known to those skilled in the art as "pro-moieties" may
be placed on appropriate functionalities when such functionalities
are present within compounds of the invention. Preferred prodrugs
for compounds of the invention include: esters, carbonate esters,
hemi-esters, phosphate esters, nitro esters, sulfate esters,
sulfoxides, amides, carbamates, azo-compounds, phosphamides,
glycosides, ethers, acetals and ketals.
[0064] Pharmaceutical compositions may be presented in unit dose
forms containing a predetermined amount of active ingredient per
unit dose. Such a unit may contain, for example, 0.5 mg to 3500 mg,
preferably 1 mg to 700 mg, more preferably 5 mg to 100 mg of a
compound of the formula (I), depending on the condition being
treated, the route of administration and the age, weight and
condition of the patient, or pharmaceutical compositions may be
presented in unit dose forms containing a predetermined amount of
active ingredient per unit dose. Preferred unit dosage compositions
are those containing a daily dose or sub-dose, as herein above
recited, or an appropriate fraction thereof, of an active
ingredient. Furthermore, such pharmaceutical compositions may be
prepared by any of the methods well known in the pharmacy art.
[0065] Pharmaceutical compositions may be adapted for
administration by any appropriate route, for example by the oral
(including buccal or sublingual), rectal, nasal, topical (including
buccal, sublingual or transdermal), vaginal or parenteral
(including subcutaneous, intramuscular, intravenous or intradermal)
route. Such compositions may be prepared by any method known in the
art of pharmacy, for example by bringing into association a
compound of formal (I) with the carrier(s) or excipient(s).
[0066] Pharmaceutical compositions adapted for oral administration
may be presented as discrete units such as capsules or tablets;
powders or granules; solutions or suspensions in aqueous or
non-aqueous liquids; edible foams or whips; or oil-in-water liquid
emulsions or water-in-oil liquid emulsions.
[0067] Capsules are made by preparing a powder mixture, as
described above, and filling formed gelatin sheaths. Glidants and
lubricants such as colloidal silica, talc, magnesium stearate,
calcium stearate or solid polyethylene glycol can be added to the
powder mixture before the filling operation. A disintegrating or
solubilizing agent such as agar-agar, calcium carbonate or sodium
carbonate can also be added to improve the availability of the
medicament when the capsule is ingested.
[0068] Moreover, when desired or necessary, suitable binders,
lubricants, disintegrating agents and coloring agents can also be
incorporated into the mixture. Suitable binders include starch,
gelatin, natural sugars such as glucose or beta-lactose, corn
sweeteners, natural and synthetic gums such as acacia, tragacanth
or sodium alginate, carboxymethylcellulose, polyethylene glycol,
waxes and the like. Lubricants used in these dosage forms include
sodium oleate, sodium stearate, magnesium stearate, sodium
benzoate, sodium acetate, sodium chloride and the like.
Disintegrators include, without limitation, starch, methyl
cellulose, agar, bentonite, xanthan gum and the like. Tablets are
formulated, for example, by preparing a powder mixture, granulating
or slugging, adding a lubricant and disintegrant and pressing into
tablets. A powder mixture is prepared by mixing the compound,
suitably comminuted, with a diluent or base as described above, and
optionally, with a binder such as carboxymethylcellulose, an
aliginate, gelatin, or polyvinyl pyrrolidone, a solution retardant
such as paraffin, a resorption accelerator such as a quaternary
salt and/or an absorption agent such as bentonite, kaolin or
dicalcium phosphate. The powder mixture can be granulated by tablet
forming dies by means of the addition of stearic acid, a stearate
salt, talc or mineral oil. The lubricated mixture is then
compressed into tablets. The compounds of the present invention can
also be combined with a free flowing inert carrier and compressed
into tablets directly without going through the granulating or
slugging steps. A clear or opaque protective coating consisting of
a sealing coat of shellac, a coating of sugar or polymeric material
and a polish coating of wax can be provided. Dyestuffs can be added
to these coatings to distinguish different unit dosages.
[0069] Oral fluids such as solution, syrups and elixirs can be
prepared in dosage unit form so that a given quantity contains a
predetermined amount of a compound of formula (I). Syrups can be
prepared by dissolving the compound in a suitably flavored aqueous
solution, while elixirs are prepared through the use of a non-toxic
alcoholic vehicle. Suspensions can be formulated by dispersing the
compound in a non-toxic vehicle. Solubilizers and emulsifiers such
as ethoxylated isostearyl alcohols and polyoxy ethylene sorbitol
ethers, preservatives, flavor additive such as peppermint oil or
natural sweeteners or saccharin or other artificial sweeteners, and
the like can also be added.
[0070] Where appropriate, dosage unit pharmaceutical compositions
for oral administration can be microencapsulated. The formulation
can also be prepared to prolong or sustain the release as for
example by coating or embedding particulate material in polymers,
wax or the like.
[0071] Pharmaceutical formulations adapted for parenteral
administration include aqueous and non-aqueous sterile injection
solutions which may contain anti-oxidants, buffers, bacteriostats
and solutes which render the composition isotonic with the blood of
the intended recipient; and aqueous and non-aqueous sterile
suspensions which may include suspending agents and thickening
agents. The pharmaceutical compositions may be presented in
unit-dose or multi-dose containers, for example sealed ampoules and
vials, and may be stored in a freeze-dried (lyophilized) condition
requiring only the addition of the sterile liquid carrier, for
example water for injections, immediately prior to use.
Extemporaneous injection solutions and suspensions may be prepared
from sterile powders, granules and tablets.
[0072] A therapeutically effective amount of a compound of the
present invention will depend upon a number of factors including,
for example, the age and weight of the intended recipient, the
precise condition requiring treatment and its severity, the nature
of the formulation, and the route of administration, and will
ultimately be at the discretion of the attendant prescribing the
medication. However, an effective amount of a compound of formula
(I) for the treatment of cancer will generally be in the range of
0.001 to 100 mg/kg body weight of recipient per day, suitably in
the range of 0.01 to 10 mg/kg body weight per day. For a 70 kg
adult the actual amount per day would suitably be from 7 to 700 mg
and this amount may be given in a single dose per day or in a
number (such as two, three, four, five or six) of sub-doses per day
such that the total daily dose is the same. An effective amount of
a salt or solvate, etc., may be determined as a proportion of the
effective amount of the compound of formula (I) per se. It is
envisaged that similar dosages would be appropriate for treatment
of the other conditions referred to above.
[0073] Treatments
[0074] The compounds and compositions of the invention are used to
treat cellular proliferation diseases. Disease states which can be
treated by the methods and compositions provided herein include,
but are not limited to, cancer, autoimmune disease, fungal
disorders, arthritis, graft rejection, inflammatory bowel disease,
proliferation induced after medical procedures, including, but not
limited to, surgery, angioplasty, and the like. It is appreciated
that in some cases the cells may not be in a hyper or hypo
proliferation state (abnormal state) and still requires treatment.
For example, during wound healing, the cells may be proliferating
"normally", but proliferation enhancement may be desired. Thus, in
one embodiment, the invention herein includes application to cells
or individuals afflicted or impending affliction with any one of
these disorders or states. These compounds may also be used for
treating macular degeration associated with neovacularization, such
as AMD
[0075] The compositions and methods provided herein are
particularly deemed useful for the treatment of cancer including
tumors such as skin, breast, brain, cervical carcinomas, testicular
carcinomas, etc. They are particularly useful in treating
metastatic or malignant tumors. More particularly, cancers that may
be treated by the compositions and methods of the invention
include, but are not limited to tumor types such as astrocytic,
breast, cervical, colorectal, endometrial, esophageal, gastric,
head and neck, hepatocellular, laryngeal, lung, oral, ovarian,
prostate and thyroid carcinomas and sarcomas. More specifically,
these compounds can be used to treat: Cardiac: sarcoma
(angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma),
myxoma, rhabdomyoma, fibroma, lipoma and teratoma; Lung:
bronchogenic carcinoma (squamous cell, undifferentiated small cell,
undifferentiated large cell, adenocarcinoma), alveolar
(bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma,
chondromatous hamartoma, mesothelioma; Gastrointestinal: esophagus
(squamous cell carcinoma, adenocarcinoma, leiomyosarcoma,
lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas
(ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma,
carcinoid tumors, vipoma), small bowel (adenocarcinoma, lymphoma,
carcinoid tumors, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma,
neurofibroma, fibroma), large bowel (adenocarcinoma, tubular
adenoma, villous adenoma, hamartoma, leiomyoma); Genitourinary
tract: kidney (adenocarcinoma, Wilm's tumor (nephroblastoma),
lymphoma, leukemia), bladder and urethra (squamous cell carcinoma,
transitional cell carcinoma, adenocarcinoma), prostate
(adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal
carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial
cell carcinoma, fibroma, fibroadenoma, adenomatoid tumors, lipoma);
Liver: hepatoma (hepatocellular carcinoma), cholangiocarcinoma,
hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma;
Bone: osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant
fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant
lymphoma (reticulum cell sarcoma), multiple myeloma, malignant
giant cell tumor chordoma, osteochronfroma (osteocartilaginous
exostoses), benign chondroma, chondroblastoma, chondromyxofibroma,
osteoid osteoma and giant cell tumors; Nervous system: skull
(osteoma, hemangioma, granuloma, xanthoma, osteitis deformans),
meninges (meningioma, meningiosarcoma, gliomatosis), brain
(astrocytoma, medulloblastoma, glioma, ependymoma, germinoma
(pinealoma), glioblastoma multiform, oligodendroglioma, schwannoma,
retinoblastoma, congenital tumors), spinal cord neurofibroma,
meningioma, glioma, sarcoma); Gynecological: uterus (endometrial
carcinoma), cervix (cervical carcinoma, pre-tumor cervical
dysplasia), ovaries (ovarian carcinoma (serous cystadenocarcinoma,
mucinous cystadenocarcinoma, unclassified carcinoma),
granulosa-thecal cell tumors, Sertoli-Leydig cell tumors,
dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma,
intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma),
vagina (clear cell carcinoma, squamous cell carcinoma, botryoid
sarcoma (embryonal rhabdomyosarcoma), fallopian tubes (carcinoma);
Hematologic: blood (myeloid leukemia (acute and chronic), acute
lymphoblastic leukemia, chronic lymphocytic leukemia,
myeloproliferative diseases, multiple myeloma, myelodysplastic
syndrome), Hodgkin's disease, non-Hodgkin's lymphoma (malignant
lymphoma); Skin: malignant melanoma, basal cell carcinoma, squamous
cell carcinoma, Kaposi's sarcoma, moles dysplastic nevi, lipoma,
angioma, dermatofibroma, keloids, psoriasis; and Adrenal glands:
neuroblastoma. Thus, the term "cancerous cell" as provided herein,
includes a cell afflicted by any one or related of the above
identified conditions.
[0076] In comparison to related 2,4-diaminopyridine derivatives
described elsewhere, the compounds of the present invention contain
a hydroxamic acid ester function on the 4-aminophenyl ring at the
2-position and an aminopyrazole at 2-position on the pyridine ring.
The hydroxamic acid ester function on the phenyl ring, as compared
with the corresponding amide, increases potency against FAK on the
order of 2-5 fold, particularly in vitro, and improves selectivity
for FAK over other enzymes. The pyrazole reduces reactivity in the
cytochrome P450s. Hence the combination of the hydroxamic acid
ester construct on the phenyl ring with an aminopyrazole at 2
position on the pyridine ring provides compounds with enhanced
safety and efficacy over other FAK inhibitors such as the
2,4-diaminopyridine derivatives.
[0077] The instant compounds can be combined with or
co-administered with other therapeutic agents, particularly agents
that may enhance the activity or time of disposition of the
compounds. Combination therapies according to the invention
comprise the administration of at least one compound of the
invention and the use of at least one other treatment method. In
one embodiment, combination therapies according to the invention
comprise the administration of at least one compound of the
invention and surgical therapy. In one embodiment, combination
therapies according to the invention comprise the administration of
at least one compound of the invention and radiotherapy. In one
embodiment, combination therapies according to the invention
comprise the administration of at least one compound of the
invention and at least one supportive care agent (e.g., at least
one anti-emetic agent). In one embodiment, combination therapies
according to the present invention comprise the administration of
at least one compound of the invention and at least one other
chemotherapeutic agent. In one particular embodiment, the invention
comprises the administration of at least one compound of the
invention and at least one anti-neoplastic agent. In yet another
embodiment, the invention comprises a therapeutic regimen where the
FAK inhibitors of this disclosure are not in and of themselves
active or significantly active, but when combined with another
therapy, which may or may not be active as a stand alone therapy,
the combination provides a useful therapeutic outcome.
[0078] By the term "co-administering" and derivatives thereof as
used herein is meant either simultaneous administration or any
manner of separate sequential administration of an FAK inhibiting
compound, as described herein, and a further active ingredient or
ingredients, known to be useful in the treatment of cancer,
including chemotherapy and radiation treatment. The term further
active ingredient or ingredients, as used herein, includes any
compound or therapeutic agent known to or that demonstrates
advantageous properties when administered to a patient in need of
treatment for cancer. Preferably, if the administration is not
simultaneous, the compounds are administered in a close time
proximity to each other. Furthermore, it does not matter if the
compounds are administered in the same dosage form, e.g. one
compound may be administered topically and another compound may be
administered orally.
[0079] Typically, any anti-neoplastic agent that has activity
versus a susceptible tumor being treated may be co-administered in
the treatment of specified cancers in the present invention.
Examples of such agents can be found in Cancer Principles and
Practice of Oncology by V. T. Devita and S. Hellman (editors),
6.sup.th edition (Feb. 15, 2001), Lippincott Williams & Wilkins
Publishers. A person of ordinary skill in the art would be able to
discern which combinations of agents would be useful based on the
particular characteristics of the drugs and the cancer involved.
Typical anti-neoplastic agents useful in the present invention
include, but are not limited to, anti-microtubule agents such as
diterpenoids and vinca alkaloids; platinum coordination complexes;
alkylating agents such as nitrogen mustards, oxazaphosphorines,
alkylsulfonates, nitrosoureas, and triazenes; antibiotic agents
such as anthracycline, actinomycins and bleomycins; topoisomerase
II inhibitors such as epipodophyllotoxins; antimetabolites such as
purine and pyrimidine analogues and anti-folate compounds;
topoisomerase I inhibitors such as camptothecins; hormones and
hormonal analogues; signal transduction pathway inhibitors;
non-receptor tyrosine kinase angiogenesis inhibitors;
immunotherapeutic agents; proapoptotic agents; and cell cycle
signaling inhibitors.
[0080] Typically, any chemotherapeutic agent that has activity
against a susceptible neoplasm being treated may be utilized in
combination with the compounds the invention, provided that the
particular agent is clinically compatible with therapy employing a
compound of the invention. Typical anti-neoplastic agents useful in
the present invention include, but are not limited to: alkylating
agents, anti-metabolites, antitumor antibiotics, antimitotic
agents, topoisomerase I and II inhibitors, hormones and hormonal
analogues; retinoids, signal transduction pathway inhibitors
including inhibitors of cell growth or growth factor function,
angiogenesis inhibitors, and serine/threonine or other kinase
inhibitors; cyclin dependent kinase inhibitors; antisense therapies
and immunotherapeutic agents, including monoclonals, vaccines or
other biological agents.
[0081] Signal transduction pathway inhibitors are those inhibitors
which block or inhibit a chemical process which evokes an
intracellular change. As used herein this change is cell
proliferation or differentiation or survival. Signal transduction
pathway inhibitors useful in the present invention include, but are
not limited to, inhibitors of receptor tyrosine kinases,
non-receptor tyrosine kinases, SH2/SH3 domain blockers,
serine/threonine kinases, phosphatidyl inositol-3-OH kinases,
myoinositol signaling, and Ras oncogenes. Signal transduction
pathway inhibitors may be employed in combination with the
compounds of the invention in the compositions and methods
described above.
[0082] Receptor kinase angiogenesis inhibitors may also find use in
the present invention. Inhibitors of angiogenesis related to VEGFR
and TIE-2 are discussed above in regard to signal transduction
inhibitors (both are receptor tyrosine kinases). Other inhibitors
may be used in combination with the compounds of the invention. For
example, anti-VEGF antibodies, which do not recognize VEGFR (the
receptor tyrosine kinase), but bind to the ligand; small molecule
inhibitors of integrin (alpha.sub.v beta.sub.3) that inhibit
angiogenesis; endostatin and angiostatin (non-RTK) may also prove
useful in combination with the compounds of the invention. One
example of a VEGFR antibody is bevacizumab) (AVASTIN.RTM.).
[0083] Several inhibitors of growth factor receptors are under
development and include ligand antagonists, antibodies, tyrosine
kinase inhibitors, anti-sense oligonucleotides and aptamers. Any of
these growth factor receptor inhibitors may be employed in
combination with the compounds of the invention in any of the
compositions and methods/uses described herein. Trastuzumab
(Herceptin.RTM.) is an example of an anti-erbB2 antibody inhibitor
of growth factor function. One example of an anti-erbB1 antibody
inhibitor of growth factor function is cetuximab (Erbitux.TM.,
C225). Bevacizumab (Avastin.RTM.) is an example of a monoclonal
antibody directed against VEGFR. Examples of small molecule
inhibitors of epidermal growth factor receptors include but are not
limited to lapatinib (Tykerb.TM.) and erlotinib (TARCEVA.RTM.).
Imatinib mesylate (GLEEVEC.RTM.) is one example of a PDGFR
inhibitor. Examples of VEGFR inhibitors include pazopanib, ZD6474,
AZD2171, PTK787, sunitinib and sorafenib. Pazopanib and the
compounds of Formula I an their salts are of particular
interest.
[0084] Anti-microtubule or anti-mitotic agents are phase specific
agents active against the microtubules of tumor cells during M or
the mitosis phase of the cell cycle. Examples of anti-microtubule
agents include, but are not limited to, diterpenoids and vinca
alkaloids.
[0085] Diterpenoids, which are derived from natural sources, are
phase specific anti-cancer agents that operate at the G.sub.2/M
phases of the cell cycle. It is believed that the diterpenoids
stabilize the .beta.-tubulin subunit of the microtubules, by
binding with this protein. Disassembly of the protein appears then
to be inhibited with mitosis being arrested and cell death
following. Examples of diterpenoids include, but are not limited
to, paclitaxel and its analog docetaxel.
[0086] Paclitaxel,
5.beta.,20-epoxy-1,2.alpha.,4,7.beta.,10.beta.,13.alpha.-hexa-hydroxytax--
11-en-9-one 4,10-diacetate 2-benzoate 13-ester with
(2R,3S)--N-benzoyl-3-phenylisoserine; is a natural diterpene
product isolated from the Pacific yew tree Taxus brevifolia and is
commercially available as an injectable solution TAXOL.RTM.. It is
a member of the taxane family of terpenes. It was first isolated in
1971 by Wani et al. J. Am. Chem, Soc., 93:2325. 1971), who
characterized its structure by chemical and X-ray crystallographic
methods. One mechanism for its activity relates to paclitaxel's
capacity to bind tubulin, thereby inhibiting cancer cell growth.
Schiff et al., Proc. Natl, Acad, Sci. USA, 77:1561-1565 (1980);
Schiff et al., Nature, 277:665-667 (1979); Kumar, J. Biol, Chem,
256: 10435-10441 (1981). For a review of synthesis and anticancer
activity of some paclitaxel derivatives see: D. G. I. Kingston et
al., Studies in Organic Chemistry vol. 26, entitled "New trends in
Natural Products Chemistry 1986", Attaur-Rahman, P. W. Le Quesne,
Eds. (Elsevier, Amsterdam, 1986) pp 219-235.
[0087] Paclitaxel has been approved for clinical use in the
treatment of refractory ovarian cancer in the United States
(Markman et al., Yale Journal of Biology and Medicine, 64:583,
1991; McGuire et al., Ann. lntem, Med., 111:273, 1989) and for the
treatment of breast cancer (Holmes et al., J. Nat. Cancer Inst.,
83:1797, 1991.) It is a potential candidate for treatment of
neoplasms in the skin (Einzig et. al., Proc. Am. Soc. Clin. Oncol.,
20:46) and head and neck carcinomas (Forastire et. al., Sem.
Oncol., 20:56, 1990). The compound also shows potential for the
treatment of polycystic kidney disease (Woo et. al., Nature,
368:750. 1994), lung cancer and malaria. Treatment of patients with
paclitaxel results in bone marrow suppression (multiple cell
lineages, Ignoff, R. J. et. al, Cancer Chemotherapy Pocket Guide,
1998) related to the duration of dosing above a threshold
concentration (50 nM) (Kearns, C. M. et. al., Seminars in Oncology,
3(6) p. 16-23, 1995).
[0088] Docetaxel, (2R,3S)--
N-carboxy-3-phenylisoserine,N-tert-butyl ester, 13-ester with
5.beta.-20-epoxy-1,2.alpha.,4,7.beta.,10.beta.,13.alpha.-hexahydroxytax-1-
1-en-9-one 4-acetate 2-benzoate, trihydrate; is commercially
available as an injectable solution as TAXOTERE.RTM.. Docetaxel is
indicated for the treatment of breast cancer. Docetaxel is a
semisynthetic derivative of paclitaxel q.v., prepared using a
natural precursor, 10-deacetyl-baccatin III, extracted from the
needle of the European Yew tree. The dose limiting toxicity of
docetaxel is neutropenia.
[0089] Vinca alkaloids are phase specific anti-neoplastic agents
derived from the periwinkle plant. Vinca alkaloids act at the M
phase (mitosis) of the cell cycle by binding specifically to
tubulin. Consequently, the bound tubulin molecule is unable to
polymerize into microtubules. Mitosis is believed to be arrested in
metaphase with cell death following. Examples of vinca alkaloids
include, but are not limited to, vinblastine, vincristine, and
vinorelbine.
[0090] Vinblastine, vincaleukoblastine sulfate, is commercially
available as VELBAN.RTM. as an injectable solution. Although, it
has possible indication as a second line therapy of various solid
tumors, it is primarily indicated in the treatment of testicular
cancer and various lymphomas including Hodgkin's Disease; and
lymphocytic and histiocytic lymphomas. Myelosuppression is the dose
limiting side effect of vinblastine.
[0091] Vincristine, vincaleukoblastine, 22-oxo-, sulfate, is
commercially available as ONCOVIN.RTM. as an injectable solution.
Vincristine is indicated for the treatment of acute leukemias and
has also found use in treatment regimens for Hodgkin's and
non-Hodgkin's malignant lymphomas. Alopecia and neurologic effects
are the most common side effect of vincristine and to a lesser
extent myelosupression and gastrointestinal mucositis effects
Occur.
[0092] Vinorelbine,
3',4'-didehydro-4'-deoxy-C'-norvincaleukoblastine[R--(R*,R*)-2,3-dihydrox-
ybutanedioate (1:2)(salt)], commercially available as an injectable
solution of vinorelbine tartrate (NAVELBINE.RTM.), is a
semisynthetic vinca alkaloid. Vinorelbine is indicated as a single
agent or in combination with other chemotherapeutic agents, such as
cisplatin, in the treatment of various solid tumors, particularly
non-small cell lung, advanced breast, and hormone refractory
prostate cancers. Myelosuppression is the most common dose limiting
side effect of vinorelbine.
[0093] Platinum coordination complexes are non-phase specific
anti-cancer agents, which are interactive with DNA. The platinum
complexes enter tumor cells, undergo, aquation and form intra- and
interstrand crosslinks with DNA causing adverse biological effects
to the tumor. Examples of platinum coordination complexes include,
but are not limited to, cisplatin and carboplatin.
[0094] Cisplatin, cis-diamminedichloroplatinum, is commercially
available as PLATINOL.RTM. as an injectable solution. Cisplatin is
primarily indicated in the treatment of metastatic testicular and
ovarian cancer and advanced bladder cancer. The primary dose
limiting side effects of cisplatin are nephrotoxicity, which may be
controlled by hydration and diuresis, and ototoxicity.
[0095] Carboplatin, platinum, diammine
[1,1-cyclobutane-dicarboxylate(2-)-O,O'], is commercially available
as PARAPLATIN.RTM. as an injectable solution. Carboplatin is
primarily indicated in the first and second line treatment of
advanced ovarian carcinoma. Bone marrow suppression is the dose
limiting toxicity of carboplatin.
[0096] Alkylating agents are non-phase anti-cancer specific agents
and strong electrophiles. Typically, alkylating agents form
covalent linkages, by alkylation, to DNA through nucleophilic
moieties of the DNA molecule such as phosphate, amino, sulfhydryl,
hydroxyl, carboxyl, and imidazole groups. Such alkylation disrupts
nucleic acid function leading to cell death. Examples of alkylating
agents include, but are not limited to, nitrogen mustards such as
cyclophosphamide, melphalan, and chlorambucil; alkyl sulfonates
such as busulfan; nitrosoureas such as carmustine; and triazenes
such as dacarbazine.
[0097] Cyclophosphamide,
2-[bis(2-chloroethyl)amino]tetrahydro-2H-1,3,2-oxazaphosphorine
2-oxide monohydrate, is commercially available as an injectable
solution or tablets as CYTOXAN.RTM.. Cyclophosphamide is indicated
as a single agent or in combination with other chemotherapeutic
agents, in the treatment of malignant lymphomas, multiple myeloma,
and leukemias. Alopecia, nausea, vomiting and leukopenia are the
most common dose limiting side effects of cyclophosphamide.
[0098] Melphalan, 4-[bis(2-chloroethyl)amino]-L-phenylalanine, is
commercially available as an injectable solution or tablets as
ALKERAN.RTM.. Melphalan is indicated for the palliative treatment
of multiple myeloma and non-resectable epithelial carcinoma of the
ovary. Bone marrow suppression is the most common dose limiting
side effect of melphalan.
[0099] Chlorambucil, 4-[bis(2-chloroethyl)amino]benzenebutanoic
acid, is commercially available as LEUKERAN.RTM. tablets.
Chlorambucil is indicated for the palliative treatment of chronic
lymphatic leukemia, and malignant lymphomas such as lymphosarcoma,
giant follicular lymphoma, and Hodgkin's disease. Bone marrow
suppression is the most common dose limiting side effect of
chlorambucil.
[0100] Busulfan, 1,4-butanediol dimethanesulfonate, is commercially
available as MYLERAN.RTM. TABLETS. Busulfan is indicated for the
palliative treatment of chronic myelogenous leukemia. Bone marrow
suppression is the most common dose limiting side effects of
busulfan.
[0101] Carmustine, 1,3-[bis(2-chloroethyl)-1-nitrosourea, is
commercially available as single vials of lyophilized material as
BiCNU.RTM.. Carmustine is indicated for the palliative treatment as
a single agent or in combination with other agents for brain
tumors, multiple myeloma, Hodgkin's disease, and non-Hodgkin's
lymphomas. Delayed myelosuppression is the most common dose
limiting side effects of carmustine.
[0102] Dacarbazine,
5-(3,3-dimethyl-1-triazeno)-imidazole-4-carboxamide, is
commercially available as single vials of material as
DTIC-Dome.RTM.. Dacarbazine is indicated for the treatment of
metastatic malignant melanoma and in combination with other agents
for the second line treatment of Hodgkin's Disease. Nausea,
vomiting, and anorexia are the most common dose limiting side
effects of dacarbazine.
[0103] Antibiotic anti-neoplastics are non-phase specific agents,
which bind or intercalate with DNA. Typically, such action results
in stable DNA complexes or strand breakage, which disrupts ordinary
function of the nucleic acids leading to cell death. Examples of
antibiotic anti-neoplastic agents include, but are not limited to,
actinomycins such as dactinomycin, anthrocyclins such as
daunorubicin and doxorubicin; and bleomycins.
[0104] Dactinomycin, also know as Actinomycin D, is commercially
available in injectable form as COSMEGEN.RTM.. Dactinomycin is
indicated for the treatment of Wilm's tumor and rhabdomyosarcoma.
Nausea, vomiting, and anorexia are the most common dose limiting
side effects of dactinomycin.
[0105] Daunorubicin,
(8S-cis-)-8-acetyl-10-[(3-amino-2,3,6-trideoxy-.alpha.-L-lyxo-hexopyranos-
yl)oxy]-7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12naphthacenedio-
ne hydrochloride, is commercially available as a liposomal
injectable form as DAUNOXOME.RTM. or as an injectable as
CERUBIDINE.RTM.. Daunorubicin is indicated for remission induction
in the treatment of acute nonlymphocytic leukemia and advanced HIV
associated Kaposi's sarcoma. Myelosuppression is the most common
dose limiting side effect of daunorubicin.
[0106] Doxorubicin,
(8S,10S)-10-[(3-amino-2,3,6-trideoxy-.alpha.-L-lyxo-hexopyranosyl)oxy]-8--
glycoloyl, 7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12
naphthacenedione hydrochloride, is commercially available as an
injectable form as RUBEX.RTM. or ADRIAMYCIN RDF.RTM.. Doxorubicin
is primarily indicated for the treatment of acute lymphoblastic
leukemia and acute myeloblastic leukemia, but is also a useful
component in the treatment of some solid tumors and lymphomas.
Myelosuppression is the most common dose limiting side effect of
doxorubicin.
[0107] Bleomycin, a mixture of cytotoxic glycopeptide antibiotics
isolated from a strain of Streptomyces verticillus, is commercially
available as BLENOXANE.RTM.. Bleomycin is indicated as a palliative
treatment, as a single agent or in combination with other agents,
of squamous cell carcinoma, lymphomas, and testicular carcinomas.
Pulmonary and cutaneous toxicities are the most common dose
limiting side effects of bleomycin.
[0108] Topoisomerase II inhibitors include, but are not limited to,
epipodophyllotoxins.
[0109] Epipodophyllotoxins are phase specific anti-neoplastic
agents derived from the mandrake plant. Epipodophyllotoxins
typically affect cells in the S and G.sub.2 phases of the cell
cycle by forming a ternary complex with topoisomerase II and DNA
causing DNA strand breaks. The strand breaks accumulate and cell
death follows. Examples of epipodophyllotoxins include, but are not
limited to, etoposide and teniposide.
[0110] Etoposide, 4'-demethyl-epipodophyllotoxin
9[4,6-0-(R)-ethylidene-.beta.-D-glucopyranoside], is commercially
available as an injectable solution or capsules as VePESID.RTM. and
is commonly known as VP-16. Etoposide is indicated as a single
agent or in combination with other chemotherapy agents in the
treatment of testicular and non-small cell lung cancers.
Myelosuppression is the most common side effect of etoposide. The
incidence of leucopenia tends to be more severe than
thrombocytopenia.
[0111] Teniposide, 4'-demethyl-epipodophyllotoxin
9[4,6-0-(R)-thenylidene-13-D-glucopyranoside], is commercially
available as an injectable solution as VUMON.RTM. and is commonly
known as VM-26. Teniposide is indicated as a single agent or in
combination with other chemotherapy agents in the treatment of
acute leukemia in children. Myelosuppression is the most common
dose limiting side effect of teniposide. Teniposide can induce both
leucopenia and thrombocytopenia.
[0112] Antimetabolite neoplastic agents are phase specific
anti-neoplastic agents that act at S phase (DNA synthesis) of the
cell cycle by inhibiting DNA synthesis or by inhibiting purine or
pyrimidine base synthesis and thereby limiting DNA synthesis.
Consequently, S phase does not proceed and cell death follows.
Examples of antimetabolite anti-neoplastic agents include, but are
not limited to, fluorouracil, methotrexate, cytarabine,
mecaptopurine, thioguanine, and gemcitabine.
[0113] 5-fluorouracil, 5-fluoro-2,4-(1H,3H) pyrimidinedione, is
commercially available as fluorouracil. Administration of
5-fluorouracil leads to inhibition of thymidylate synthesis and is
also incorporated into both RNA and DNA. The result typically is
cell death. 5-fluorouracil is indicated as a single agent or in
combination with other chemotherapy agents in the treatment of
carcinomas of the breast, colon, rectum, stomach and pancreas.
Myelosuppression and mucositis are dose limiting side effects of
5-fluorouracil. Other fluoropyrimidine analogs include 5-fluoro
deoxyuridine (floxuridine) and 5-fluorodeoxyuridine
monophosphate.
[0114] Cytarabine, 4-amino-1-.beta.-D-arabinofuranosyl-2
(1H)-pyrimidinone, is commercially available as CYTOSAR-U.RTM. and
is commonly known as Ara-C. It is believed that cytarabine exhibits
cell phase specificity at S-phase by inhibiting DNA chain
elongation by terminal incorporation of cytarabine into the growing
DNA chain. Cytarabine is indicated as a single agent or in
combination with other chemotherapy agents in the treatment of
acute leukemia. Other cytidine analogs include 5-azacytidine and
2',2'-difluorodeoxycytidine (gemcitabine). Cytarabine induces
leucopenia, thrombocytopenia, and mucositis.
[0115] Mercaptopurine, 1,7-dihydro-6H-purine-6-thione monohydrate,
is commercially available as PURINETHOL.RTM.. Mercaptopurine
exhibits cell phase specificity at S-phase by inhibiting DNA
synthesis by an as of yet unspecified mechanism. Mercaptopurine is
indicated as a single agent or in combination with other
chemotherapy agents in the treatment of acute leukemia.
Myelosuppression and gastrointestinal mucositis are expected side
effects of mercaptopurine at high doses. A useful mercaptopurine
analog is azathioprine.
[0116] Thioguanine, 2-amino-1,7-dihydro-6H-purine-6-thione, is
commercially available as TABLOID.RTM.. Thioguanine exhibits cell
phase specificity at S-phase by inhibiting DNA synthesis by an as
of yet unspecified mechanism. Thioguanine is indicated as a single
agent or in combination with other chemotherapy agents in the
treatment of acute leukemia. Myelosuppression, including
leucopenia, thrombocytopenia, and anemia, is the most common dose
limiting side effect of thioguanine administration. However,
gastrointestinal side effects occur and can be dose limiting. Other
purine analogs include pentostatin, erythrohydroxynonyladenine,
fludarabine phosphate, and cladribine.
[0117] Gemcitabine, 2'-deoxy-2',2'-difluorocytidine
monohydrochloride (.beta.-isomer), is commercially available as
GEMZAR.RTM.. Gemcitabine exhibits cell phase specificity at S-phase
and by blocking progression of cells through the G1/S boundary.
Gemcitabine is indicated in combination with cisplatin in the
treatment of locally advanced non-small cell lung cancer and alone
in the treatment of locally advanced pancreatic cancer.
Myelosuppression, including leucopenia, thrombocytopenia, and
anemia, is the most common dose limiting side effect of gemcitabine
administration.
[0118] Methotrexate,
N-[4[[(2,4-diamino-6-pteridinyl)methyl]methylamino]benzoyl]-L-glutamic
acid, is commercially available as methotrexate sodium.
Methotrexate exhibits cell phase effects specifically at S-phase by
inhibiting DNA synthesis, repair and/or replication through the
inhibition of dyhydrofolic acid reductase which is required for
synthesis of purine nucleotides and thymidylate. Methotrexate is
indicated as a single agent or in combination with other
chemotherapy agents in the treatment of choriocarcinoma, meningeal
leukemia, non-Hodgkin's lymphoma, and carcinomas of the breast,
head, neck, ovary and bladder. Myelosuppression (leucopenia,
thrombocytopenia, and anemia) and mucositis are expected side
effect of methotrexate administration.
[0119] Camptothecins, including, camptothecin and camptothecin
derivatives are available or under development as Topoisomerase I
inhibitors. Camptothecins cytotoxic activity is believed to be
related to its Topoisomerase I inhibitory activity. Examples of
camptothecins include, but are not limited to irinotecan,
topotecan, and the various optical forms of
7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20-camptoth-
ecin described below.
[0120] Irinotecan HCl,
(4S)-4,11-diethyl-4-hydroxy-9-[(4-piperidinopiperidino)
carbonyloxy]-1H-pyrano[3',4',6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)--
dione hydrochloride, is commercially available as the injectable
solution CAMPTOSAR.RTM..
[0121] Irinotecan is a derivative of camptothecin which binds,
along with its active metabolite SN-38, to the topoisomerase I-DNA
complex. It is believed that cytotoxicity occurs as a result of
irreparable double strand breaks caused by interaction of the
topoisomerase I:DNA:irintecan or SN-38 ternary complex with
replication enzymes. Irinotecan is indicated for treatment of
metastatic cancer of the colon or rectum. The dose limiting side
effects of irinotecan HCl are myelosuppression, including
neutropenia, and GI effects, including diarrhea.
[0122] Topotecan HCl,
(S)-10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy-1H-pyrano[3',4',6,7]-
indolizino[1,2-b]quinoline-3,14-(4H,12H)-dione monohydrochloride,
is commercially available as the injectable solution HYCAMTIN.RTM..
Topotecan is a derivative of camptothecin which binds to the
topoisomerase I-DNA complex and prevents religation of singles
strand breaks caused by Topoisomerase I in response to torsional
strain of the DNA molecule. Topotecan is indicated for second line
treatment of metastatic carcinoma of the ovary and small cell lung
cancer. The dose limiting side effect of topotecan HCl is
myelosuppression, primarily neutropenia.
[0123] The following schemes illustrate how compounds of the
present invention can be prepared. The specific solvents and
reaction conditions referred to are also illustrative and are not
intended to be limiting.
Schemes Compounds of formula (I) may be prepared by the methods
outlined in Scheme 1 below. Compounds of formula (II) and (III) are
commercially available or may be synthesized using techniques
conventional in the art. The L group for compound (III) represents
a leaving group such as F or Cl. The compounds of formula (II) and
(III) may be reacted under reflux or microwave conditions to afford
intermediate (IV). The addition reaction is typically done using a
polar, protic solvents such as n-butanol or iso-propanol.
Alternatively, metal catalyzed coupling reaction conditions may be
used. When compound (II) includes a functional group in need of
protection, for example, a hydroxyl or amino group, an appropriate
protecting group is advantageously used. Compounds of formula (IV)
may then be reacted with an aminopyrazole (V), which is
commercially available or which may be synthesized using techniques
conventional in the art, to afford a compound of formula (I). The
reaction is typically carried out in the presence of a metal
catalyst, such as a palladium salt, along with an appropriate
phosphine ligand. Alternatively, the reaction can be carried out
with a catalytic amount of an acid such as hydrochloric or
trifluoroacetic acid and in a suitable solvent such as water,
1,4-dioxane, or iso-propanol or a combination thereof; the reaction
is advantageously carried out at an advanced temperature, for
example, under refluxing conditions, or by using a microwave
apparatus. The acid catalyst is typically present in an amount of
10-30 mol % with respect to the compound of formula (I).
##STR00005##
[0124] Compounds of formula (VIII) may be conveniently prepared by
the methods outlined in Scheme 1, but starting with an appropriate
anthranilamide (VI), as outlined in Scheme 2.
##STR00006##
[0125] Compound (VI) may contain additional substituents. For
example, as shown in Scheme 3, benzoxazine (IX), which is either
commercially available or synthesized using techniques conventional
in the art, can be ring-opened with an amine to form benzamide (X),
which can then undergo addition with compound (III) to yield the
compound of formula (XI).
##STR00007##
[0126] A compound of formula (XII) may be prepared by reacting a
compound of formula (II) with a compound of formula (XIII). This
reaction can be carried out as described in Scheme 1. Compounds of
formula (XII) may then be reacted with a compound of formula (XIV)
to give compounds of formula (I). The reaction may be carried out
in inert solvent, in the presence of a metal catalyst and
appropriate ligand.
##STR00008##
[0127] Certain compounds of formula (I) can also be prepared as
outlined in Scheme 5. The amino group of the compound of formula
(XV) can first be reacted with diketene followed by acylation and
treatment with a hydrazine. Compound of formula (XVI) can then be
obtained by treatment with acid, then reacted with a compound of
formula (II) to give a compound of formula (I). This last reaction
can be carried out as described in Scheme 1.
##STR00009##
[0128] Compounds of formula (V) can be made by the condensation of
a substituted hydrazine (XVIII) with the appropriate cyano-ketone
(XVII), for example, according to the procedures of Honma, T. et
al. J. Med. Chem. 2002, Vol. 44 (26), 4628-4640 or Adachi, I. et
al. Chemical & Pharmaceutical Bulletin 1987, 35(8), 3235-52 as
outlined in Scheme 6.
##STR00010##
##STR00011##
[0129] A compound of formula (XXI) can also be prepared as outlined
in Scheme 7. The nitrile of formula (XIV) can be hydrolyzed to a
carboxylic acid of formula (XX) and then coupled with an amine to
give compounds of formula (XXI).
Experimentals
Biochemical Assay for FAK Activity
[0130] Assay 1:
[0131] GST-tagged (glutathione S-transferase-tagged) FAK was
purchased from Invitrogen (PV3832) (www.invitrogen.com). The
activity of FAK was measured by monitoring the phosphorylation of a
peptide substrate (Ac-RRRRRRSETDDYAEIID-NH.sub.2; (SEQ ID NO: 1)
i.e.
Ac-Arg-Arg-Arg-Arg-Arg-Ser-Glu-Thr-Asp-Asp-Tyr-Ala-Glu-Ile-Ile-Asp-NH.sub-
.2) in the presence of a radio-labeled ATP. To measure inhibitors
of FAK, compounds were first prepared as a 10.times. stock in 10%
DMSO. A small portion of each solution (4 .mu.L) was added to a
96-well plate (Corning, 3884). A 6-nM GST-FAK solution was prepared
in 1.1.times. reaction buffer containing 44 mM HEPES, pH=7.2, 11 mM
MgCl.sub.2, 2.2 mM MnCl.sub.2, 1.1 mM DTT and 0.011% Tween-20.
Then, 20 .mu.L of the 6 nM GST-FAK solution were pre-incubated with
the compounds for 30 min at room temperature. The reaction was
initiated by adding 16 .mu.L of substrates (62.5 .mu.M peptide, 5
.mu.M ATP and .about.0.02 mCi/mL .sup.33P-.gamma.-ATP) prepared in
the above reaction buffer. The reaction was allowed to proceed for
90 min before being quenched with 40 .mu.L of 1% H.sub.3PO.sub.4. A
portion of the reaction mixture (60 .mu.L) was transferred to a
phospho-cellulose filter plate (Millipore; www.millipore.com,
MAPHNOB50) and incubated for 20 minutes. The plate was filtrated,
washed three times using 150 .mu.L of 0.5% H.sub.3PO.sub.4 and
dried at 50.degree. C. for 30 min. After the addition of 60 .mu.L
Microscint-20 to the plate, radioactivity was measured using a
TopCount (PerkinElmer; www.PerkinElmer.com).
[0132] Assay 2:
[0133] Flag-His-TEV-FAK1 was prepared in-house. Full length Human
FAK was expressed using baculovirus in Sf9 cells with N-terminal
FLAG-6.times. His tags followed by a TEV cleavage site
(FLAG-6.times. His-TEV-huFAK). The activity of FAK was measured by
monitoring the phosphorylation of LANCE Ultra
NH.sub.2-(ULight)-CSETDDYAEIID-COOH (SEQ ID NO: 2) (C=cysteine
S=serine, E=glutamic acid, T=threonine, D=aspartic acid,
Y=tyrosine, A=alanine, I=isoluecine) substrate (purchased from
Perkin Elmer Life Sciences). To measure inhibitors of FAK,
compounds were first prepared as a 100.times. stock in 100% DMSO. A
small portion of each compound solution (50 nL) was added to a
black 384-well low-volume microtiter plate (Greiner 784076). A 1.2
nM Flag-His-TEV-FAK1 solution was prepared in 1.times. reaction
buffer containing 40 mM Tris/Tris-HCL, 10 mM MgCl2, 1 mM CHAPS, at
a pH of 7.5, with 1 mM DTT added. 2.5 ul of the 1.2 nM Flag-FAK
solution was added to the plates and pre-incubated with the
compounds for 30 min at room temperature. Then, 2.5 .mu.L of
substrate solution (0.1 .mu.M of P2 FAK-tide specific substrate
(Lance Ultra NH2-(ULight)-CSETDDYAEIID-COOH (SEQ ID NO: 2) from
Perkin Elmer), 10 .mu.M ATP and the 1.times. reaction buffer
described above), was added to the plate to initiate the reaction.
After incubating for 120 minutes at room temperature, the reaction
is quenched by adding 5 uL of 20 mM EDTA and 5 nMEu-Anti-pTyr
antibody in 1.times. LANCE detection buffer. After a 30 minute
incubation at room temperature, the plate is read on a Perkin Elmer
Viewlux with a 320-340 nm excitation filter and measuring emission
at 615 nm and 665 nm. The ratio of 665 nm/615 nm is used for data
normalization.
[0134] The following Table A provides specific data for compounds
of the below Examples as run in one or both of the foregoing
assays. These data were generated in at least one run in the noted
assay; repeats assay runs may have given or may give readouts that
vary to some degree from these data.
TABLE-US-00001 TABLE A Assay 2 Assay 1 FAK H Example FAK TRF 31284A
No. PXC.sub.50 TRF PXC.sub.50 1 8.7 7.8 2 8.8 8 3 9.2 8.3 4 9 8.2 5
9 8.1 6 8.9 8.3 7 9.1 8 8 9.2 8.3 9 9 8.2 10 9.1 8 11 8.7 8.2 12
8.6 8.3 13 9.3 8.7 14 8.6 7.7 15 8.6 7.7 16 8.9 7.7 17 8.9 7.4 18 8
19 8.3 7.2 20 8 7.4 21 8.5 7.2 22 7.5 6.6 23 7.1 6.7 24 8.2 7.2 25
8.6 7.5 26 9.4 8.4 27 8.7 8.1 28 9.4 8.6 29 9.3 8.7 30 9.4 8.8 31
8.4 32 9.4 9 33 9.4 34 9.3 36 9.4 8.8 37 8.4 7.7 38 6.6 6 39 9.4
8.3 40 8.9 8.6 41 9.4 8.7 42 7.9 7.3 43 8.3 7.6 44 8.2 7.2 45 8 7.2
46 7.9 47 8.6 49 7.2 50 7.9 51 7.7 52 8.2 53 7.4 54 7.5 55 8 56 7.8
58 7.3 59 9.2 8.3 60 7.9 61 7.6 62 63 7.3 64 7.1 65 7.8 66 8.7 7.5
67 7.2 8.3 69 8.6 70 8.2 71 8.2 72 8.6 73 8.9
Chemistry Examples
[0135] The following chemistry examples are for illustrative
purposes only and are not intended to limit the scope of the
present invention. The compounds were named using ACD Name software
(Advanced Chemistry Development, www.acdlabs.com). All compounds
have pIC.sub.50 of greater than 6.5 for the above-described
biochemical assay.
[0136] A PE Sciex API 150 single quadrupole mass spectrometer (PE
Sciex, Thornhill, Ontario, Canada) was operated using electrospray
ionization in the positive ion detection mode. The nebulizing gas
was generated from a zero air generator (Balston Inc., Haverhill,
Mass.; www.parker.com) and delivered at 65 psi and the curtain gas
was high purity nitrogen delivered from a Dewar liquid nitrogen
vessel at 50 psi. The voltage applied to the electrospray needle
was 4.8 kV. The orifice was set at 25 V and mass spectrometer was
scanned at a rate of 0.5 scan/sec using a step mass of 0.2 amu and
collecting profile data.
[0137] Method A, LCMS. Samples are introduced into the mass
spectrometer using a CTC PAL autosampler (LEAP Technologies,
Carrboro, N.C.) equipped with a hamilton 10 uL syringe which
performed the injection into a Valco 10-port injection valve. The
HPLC pump was a Shimadzu LC-10ADvp (Shimadzu Scientific
Instruments, Columbia, Md.) operated at 0.3 mL/min and a linear
gradient 4.5% A to 90% B in 3.2 min. with a 0.4 min. hold. The
mobile phase was composed of 100% (H.sub.2O 0.02% TFA) in vessel A
and 100% (CH.sub.3CN 0.018% TFA) in vessel B. The stationary phase
is Aquasil (C18) and the column dimensions are 1 mm.times.40 mm.
Detection was by UV at 214 nm, evaporative light-scattering (ELSD)
and MS.
[0138] Method B, LCMS. Alternatively, an Agilent 1100 analytical
HPLC system with an LC/MS was used and operated at 1 mL/min and a
linear gradient 5% A to 100% B in 2.2 min with a 0.4 min hold. The
mobile phase was composed of 100% (H.sub.2O 0.02% TFA) in vessel A
and 100% (CH.sub.3CN 0.018% TFA) in vessel B. The stationary phase
was Zobax (C8) with a 3.5 um partical size and the column
dimensions were 2.1 mm.times.50 mm. Detection was by UV at 214 nm,
evaporative light-scattering (ELSD) and MS.
[0139] Method B, LCMS. Alternatively, an MDSSCIEX API 2000 equipped
with a capillary column of (50.times.4.6 mm, 5 .mu.m) was used.
HPLC was done on Agilent-1200 series UPLC system equipped with
column Zorbax SB-C18 (50.times.4.6 mm, 1.8 .mu.m) eluting with
CH.sub.3CN: ammonium acetate buffer. The reactions were performed
in the microwave (CEM, Discover).
[0140] 1H-NMR (hereinafter "NMR") spectra were recorded at 400 MHz
using a Bruker AVANCE 400 MHz instrument, with ACD Spect manager
ver 10 using for reprocessing. Multiplicities indicated are:
s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet, dd=doublet
of doublets, dt=doublet of triplets etc. and br indicates a broad
signal.
[0141] Analytical HPLC: Products were analyzed by Agilent 1100
Analytical Chromatography system, with 4.5.times.75 mm Zorbax
XDB-C18 column (3.5 m) at 2 mL/min with a 4 min gradient from 5%
CH.sub.3CN (0.1% formic acid) to 95% CH.sub.3CN (0.1% formic acid)
in H.sub.2O (0.1% formic acid) and a 1 min hold.
[0142] Preparative HPLC: Products were purified using a Gilson
preparative chromatography system with a 75.times.30 mm I. D. YMC
CombiPrep ODS-A column (5 m) (www.waters.com) at 50 mL/min with a
10 min gradient from 5% CH.sub.3CN (0.1% formic acid) to 95%
CH.sub.3CN (0.1% formic acid) in H.sub.2O (0.1% formic acid) and a
2 min hold; alternatively, products were purified using an Agilent
1100 Preparative Chromatography system, with 100.times.30 mm Gemini
C18 column (5 m) at 60 mL/min with a 10 min gradient from 5%
CH.sub.3CN (0.1% formic acid) to 95% CH.sub.3CN (0.1% formic acid)
in H.sub.2O (0.1% formic acid) and a 2 min hold.
[0143] Preparative normal phase chromatography was carried out
using an Analogix IntelliFlash 280 System with SuperFlash Sepra Si
50 columns. Alternatively, reverse-phase HPLC was performed on
Agilent using Zorbax SB-C18 column (21.2.times.250 mm, 7 .mu.m)
eluting with CH.sub.3CN: ammonium acetate buffer (10 .mu.M) at pH
6.8.
EXAMPLES
Example 1
1a) 2-[(2,5-Dichloro-4-pyridinyl)amino]-N-methylbenzamide
##STR00012##
[0145] A 150-mL sealed tube was charged with
2,5-dichloro-4-iodopyridine (3.5 g, 12.78 mmol),
2-amino-N-methylbenzamide (1.919 g, 12.78 mmol) and tripotassium
phosphate (8.14 g, 38.3 mmol) in 1,4-dioxane (100 mL). The reaction
mixture was degassed with nitrogen for 10 min.
Bis(2-diphenylphosphinophenyl)ether (DPEPhos, 0.688 g, 1.278 mmol)
and Pd(OAc).sub.2 (0.115 g, 0.511 mmol) were added and the reaction
mixture was heated in a 120.degree. C. oil bath over night. The
reaction mixture was filtered through celite, which was washed with
dioxane. The solvent was evaporated to dryness and the solid was
washed with EtOH (10 mL.times.3) to give 2.14 g (56%) of product as
an off white solid.
1b)
2-[5-Chloro-2-(2-methyl-5-phenyl-2H-pyrazol-3-ylamino)-pyridin-4-ylami-
no]-N-methyl-benzamide
##STR00013##
[0147] A 50-mL sealed tube was charged with Pd(OAc).sub.2 (18 mg,
0.08 mmol) and 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (BINAP,
50 mg, 0.08 mmol) in 1,4-dioxane (10 mL). The mixture was degassed
using bubbling nitrogen for 40 min and heated at 50.degree. C. for
1 h. Then the mixture was cooled to room temperature whereupon
2-[(2,5-dichloro-4-pyridinyl)amino]-N-methyl-benzamide (300 mg,
1.01 mmol), 2-methyl-5-phenyl-2H-pyrazol-3-ylamine (704 mg, 4.06
mmol) and cesium carbonate (960 mg, 2.96 mmol) were added under an
inert atmosphere. The tube was sealed and heated at 120.degree. C.
overnight. The reaction mixture was concentrated under reduced
pressure and the crude product was purified by column
chromatography (silica gel, eluted with dichloromethane-methanol
(DCM-MeOH) 99:1 followed by purification by preparatory TLC to
afford the desired compound as a off white solid (25 mg, 5%).
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 2.76 (d, 3H, J=4.52
Hz), 3.68 (s, 3H), 6.69 (s, 1H), 6.85 (s, 1H), 7.07-7.14 (m, 1H),
7.25-7.31 (m, 1H), 7.36-7.42 (m, 2H), 7.46-7.53 (m, 1H), 7.63 (d,
1H, J=8.08 Hz), 7.69 (d, 1H, J=7.4 Hz), 7.75 (d, 2H, J=7.16 Hz),
8.04 (s, 1H), 8.69 (brs, 1H), 8.81 (s, 1H), 10.13 (s, 1H). LC-MS
calculated for C.sub.23H.sub.21ClN.sub.6O (M+H) 433.15. found
433.3. HPLC purity 96% at .lamda.=200 nm and 99% at .lamda.=260
nm.
Example 2
2-({5-Chloro-2-[(1-methyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amino)-N-meth-
ylbenzamide
##STR00014##
[0149] A mixture of
2-[(2,5-dichloro-4-pyridinyl)amino]-N-methylbenzamide (100 mg,
0.338 mmol), 5-amino-1-methyl-1H-pyrazole (65.6 mg, 0.675 mmol),
Cs.sub.2CO.sub.3 (220 mg, 0.675 mmol),
tris(dibenzylideneacetone)dipalladium(0) (Pd.sub.2(dba).sub.3, 61.8
mg, 0.068 mmol) and 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene
(Xantphos, 48.8 mg, 0.084 mmol) was heated at 150.degree. C. in a
microwave oven for 30 min. The reaction mixture was filtered, and
the filtrate was concentrated, and the resulting crude product was
purified by reverse phase HPLC and the product was treated with 2 N
HCl to give 34 mg of product as an HCl salt (24%). LCMS (ES)
m/z=357.1 (M+H); 1H NMR (400 MHz, methanol-d4) .delta. ppm 8.01 (s,
1H), 7.76-7.74 (m, 1H), 7.62-7.61 (m, 2H), 7.55 (d, J=2 Hz, 1H),
7.42-7.39 (m, 1H), 6.52 (s, 1H), 6.33 (d, J=2 Hz, 1H), 3.76 (s,
3H), 2.91 (s, 3H).
Example 3
2-({5-Chloro-2-[(1-ethyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amino)-N-methy-
lbenzamide
##STR00015##
[0151] The title compound was prepared substantially as described
in Example 2 except using 5-amino-1-ethyl-1H-pyrazole instead of
5-amino-1-methyl-1H-pyrazole. LCMS (ES) m/z=371.1 (M+H); 1H NMR
(400 MHz, METHANOL-d4) .delta. ppm 8.06 (m, 1H), 7.77-7.61 (m, 4H),
7.44-7.40 (m, 1H), 6.61 (m, 1H), 6.40 (s, 1H), 4.17-4.10 (m, 2H),
2.91 (s, 3H), 1.40 (tr, J=7.2 Hz, 3H).
Example 4
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyrid-
inyl)amino]-N-methylbenzamide
##STR00016##
[0153] The title compound was prepared following the procedure in
Example 2 except using 5-amino-1-isopropyl-1H-pyrazole instead of
5-amino-1-methyl-1H-pyrazole. LCMS (ES) m/z=399.2 (M+H); 1H NMR
(400 MHz, methanol-d4) .delta. ppm 8.06 (s, 1H), 7.78-7.76 (m, 1H),
7.65-7.63 (m, 2H), 7.45-7.38 (m, 1H), 6.64 (s, 1H), 6.29 (s, 1H),
4.70-4.55 (m, 1H), 2.91 (s, 3H), 2.34 (s, 1H), 1.47 (d, J=6.8 Hz,
6H).
Example 5
2-({5-Chloro-2-[(1,3-dimethyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amino)-N--
methylbenzamide
##STR00017##
[0155] The title compound was prepared following the procedure in
Example 2 except using 5-amino-1-methyl-3-methyl-1H-pyrazole
instead of 5-amino-1-methyl-1H-pyrazole. LCMS (ES) m/z=371.1 (M+H);
1H NMR (400 MHz, methanol-d4) .delta. ppm 8.12 (s, 1H), 7.78-7.76
(d, J=7.6 Hz, 1H), 7.66-7.64 (m, 2H), 7.45-7.41 (m, 1H), 6.67 (s,
1H), 6.38 (s, 1H), 3.80 (s, 3H), 2.91 (s, 3H), 2.35 (s, 3H).
Example 6
2-({5-Chloro-2-[(3-cyclopropyl-1-methyl-1H-pyrazol-5-yl)amino]-4-pyridinyl-
}amino)-N-methylbenzamide
##STR00018##
[0157] The title compound was prepared following the procedure in
Example 2 except using 5-amino-3-cyclopropyl-1-methyl-1H-pyrazole
instead of 5-amino-1-methyl-1H-pyrazole. LCMS (ES) m/z=397.1 (M+H);
1H NMR (400 MHz, methanol-d4) .delta. ppm 8.06 (s, 1H), 7.78-7.76
(m, 1H), 7.64-7.62 (m, 2H), 7.44-7.40 (m, 1H), 6.57 (s, 1H), 6.15
(s, 1H), 3.71 (s, 3H), 2.91 (s, 3H), 1.97-1.90 (m, 1H), 1.04-1.00
(m, 2H), 0.79-0.76 (m, 2H).
Example 7
2-({5-Chloro-2-[(1,3-dimethyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amino)-5--
fluoro-N-methylbenzamide
##STR00019##
[0158] 7a) 2-Amino-5-fluoro-N-methylbenzamide
##STR00020##
[0160] 6-Fluoro-2H-3,1-benzoxazine-2,4(1H)-dione (200 mg, 1.104
mmol) was dissolved in dry tetrahydrofuran (THF) (10 mL), at which
time methyl amine (3.31 mL, 6.63 mmol) was added. The reaction was
stirred at room temperature for 1 h, then concentrated under
vacuum. The crude product was purified on silica (Biotage, 40%
EtOAc/hexene) to afford the title compound (120 mg, 65%) as a white
solid. LC-MS (ES) m/z=169.1 (M+H).sup.+
7b)
2-({5-Chloro-2-[(1,3-dimethyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amino-
)-5-fluoro-N-methylbenzamide
[0161] The title compound was prepared as a white solid by first
reacting 2-amino-5-fluoro-N-methylbenzamide with
2,5-dichloro-4-iodopyridine to form
2-[(2,5-dichloro-4-pyridinyl)amino]-5-fluoro-N-methylbenzamide
substantially according to the procedure of Intermediate 1, then
reacting this intermediate with 1,3-dimethyl-1H-pyrazol-5-amine
substantially according to the procedure of Example 2. LC-MS (ES)
m/z=389.1 (M+H).sup.+, .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta.
8.01 (s, 1H), 7.60 (m, 1H), 7.51 (m, 1H), 7.38 (m, 1H), 6.39 (s,
1H), 6.18 (s, 1H), 3.69 (s, 3H), 2.89 (s, 3H), 2.24 (s, 3H)
Example 8
2-({5-Chloro-2-[(1,3-dimethyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amino)-3--
fluoro-N-methylbenzamide
##STR00021##
[0162] 8a) 2-Amino-3-fluoro-N-methylbenzamide
##STR00022##
[0164] 2-Amino-3-fluorobenzonitrile (3.8 g, 27.9 mmol) was
dissolved in ethanol, water (15 mL) and THF (0.3 mL), potassium
hydroxide (7.83 g, 140 mmol) was added. The mixture was heated at
85.degree. C. for 12 h, cooled and filtered. The filtrate was
concentrated and the residue was dissolved in dichloromethane (DCM,
50 mL). Then diisopropylethylamine (19.50 mL, 112 mmol) was added,
followed by methyl amine (20.94 ml, 41.9 mmol) and
bromo-tris-pyrrolidino phosphoniumhexafluorophosphate (PyBrOP,
21.79 g, 41.9 mmol). The reaction was stirred at room temperature
for 2 h, washed with brine and dried over MgSO.sub.4. The solvent
was removed and the residue was purified by chromatography on
silica gel (20% EtOAc/Hex) to give 1.5 g (35% yield) of
intermediate a.
8b).
2-({5-Chloro-2-[(1,3-dimethyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amin-
o)-3-fluoro-N-methylbenzamide
[0165] The title compound was prepared as a white solid according
to the procedure of Example 7, except using
2-amino-3-fluoro-N-methylbenzamide in place of
2-amino-5-fluoro-N-methylbenzamide: LC-MS (ES) m/z=389.1
(M+H).sup.+, .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 8.01 (s,
1H), 7.50 (m, 3H), 6.14 (s, 1H), 5.87 (m, 1H), 3.65 (s, 3H), 2.89
(s, 3H), 2.22 (s, 3H)
[0166] The 2-{[5-chloro-2-(amino
pyrazole)-4-pyridinyl]amino}-benzamide compounds illustrated in
Table 1 were prepared from various
2-[(2,5-dichloro-4-pyridinyl)amino]-methylbenzamides and
amino-pyrazoles substantially according to the procedure of Example
7. In the following tables, the dashed lines indicate the points of
attachment. Thus, for Example 9, the compound corresponds to the
following structure:
##STR00023##
TABLE-US-00002 TABLE 1 ##STR00024## Ex. Name R.sub.a R.sub.b Data 9
2-({5-Chloro-2-[(1,3- dimethyl-1H-pyrazol-5- yl)amino]-4-
pyridinyl}amino)-4,5- difluoro-N- methylbenzamide ##STR00025##
##STR00026## LC-MS (ES) m/z = 407.1 (M + H).sup.+, .sup.1H NMR
(CD.sub.3OD, 400 MHz) .delta. 8.04 (s, 1H), 7.71 (m, 1H), 7.61 (m,
1H), 6.63 (s, 1H), 6.28 (s, 1H), 3.72 (s, 3H), 2.89 (s, 3H), 2.72
(s, 3H) 10 2-({5-Chloro-2-[(1-ethyl- 1H-pyrazol-5-yl)amino]-
4-pyridinyl}amino)-4,5- difluoro-N- methylbenzamide ##STR00027##
##STR00028## LC-MS (ES) m/z = 460.1 (M + H).sup.+, .sup.1H NMR
(CD.sub.3OD, 400 MHz) .delta. 8.00 (s, 1H), 7.70 (m, 1H), 7.59 (m,
2H), 6.56 (s, 1H), 6.33 (s, 1H), 4.11 (m, 2H), 2.90 (m, 3H), 1.38
(m, 3H) 11 5-Chloro-2-({5-chloro-2- [(1,3-dimethyl-1H-
pyrazol-5-yl)amino]-4- pyridinyl}amino)-N- methylbenzamide
##STR00029## ##STR00030## LC-MS (ES) m/z = 407.0 (M + H).sup.+,
.sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 8.03 (s, 1H), 7.78 (m,
1H), 7.61 (m, 2H), 6.56 (s, 1H), 6.23 (s, 1H), 3.70 (s, 3H), 2.90
(s, 3H), 2.26 (s, 3H) 12 5-Chloro-2-({5-chloro-2-
[(1-ethyl-1H-pyrazol-5- yl)amino]-4- pyridinyl}amino)-N-
methylbenzamide ##STR00031## ##STR00032## LC-MS (ES) m/z = 407.0 (M
+ H).sup.+, .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 8.00 (s, 1H),
7.78 (m, 1H), 7.59 (m, 3H), 6.50 (s, 1H), 6.32 (m, 1H), 4.10 (m,
2H), 2.89 (s, 3H), 1.37 (s, 3H) 13 2-({5-Chloro-2-[(1-ethyl-
3-methyl-1H-pyrazol-5- yl)amino]-4- pyridinyl}amino)-N-
methylbenzamide ##STR00033## ##STR00034## LCMS (ES) m/z = 384.8 (M
+ H); 1H NMR (400 MHz, METHANOL-d4) .delta. ppm 8.01 (s, 1H),
7.76-7.74 (m, 1H), 7.62-7.61 (m, 2H), 7.41-7.37 (m, 1H), 6.59 (s,
1H), 6.20 (s, 1H), 4.07-4.01 (m, 2H), 2.91 (s, 3H), 2.27 (s, 3H),
1.35 (tr, J = 7.2 Hz, 3H). 14 2-[(5-Chloro-2-{[3-(1,1-
dimethylethyl)-1-methyl- 1H-pyrazol-5-yl]amino}-
4-pyridinyl)amino]-N- methylbenzamide ##STR00035## ##STR00036##
LCMS (ES) m/z = 412.8 (M + H); 1H NMR (400 MHz, METHANOL-d4)
.delta. ppm 8.05 (s, 1H), 7.75-7.72 (m, 1H), 7.60-7.58 (m, 2H),
7.40-7.35 (m, 1H), 6.52 (s, 1H), 6.27 (s, 1H), 3.70 (s, 3H), 2.91
(s, 3H), 1.30 (s, 9H). 15 2-[(5-Chloro-2-{[1-ethyl-
3-(hydroxymethyl)-1H- pyrazol-5-yl]amino}-4- pyridinyl)amino]-N-
methylbenzamide ##STR00037## ##STR00038## LCMS (ES) m/z = 401.1 (M
+ H); 1H NMR (400 MHz, METHANOL-d4) .delta. ppm 7.99 (s, 1H),
7.75-7.73 (m, 1H), 7.61-7.59 (m, 2H), 7.39-7.35 (m, 1H), 6.58 (s,
1H), 6.31 (s, 1H), 4.56 (s, 2H), 4.08-4.03 (m, 2H), 2.91 (s, 3H),
1.36 (tr, J = 7.2 Hz, 6H). 16 2-[(5-Chloro-2-{[1-(2-
hydroxyethyl)-3-methyl- 1H-pyrazol-5-yl]amino}-
4-pyridinyl)amino]-N- methylbenzamide ##STR00039## ##STR00040##
LCMS (ES) m/z = 401.1 (M + H); 1H NMR (400 MHz, METHANOL-d4)
.delta. ppm 8.07 (s, 1H), 7.78-7.76 (m, 1H), 7.65-7.63 (m, 2H),
7.43-7.39 (m, 1H), 6.69 (s, 1H), 6.31 (s, 1H), 4.18 (tr, J = 5.2
Hz, 2H), 3.85 (tr, J = 5.2 Hz, 2H), 2.91 (s, 3H), 2.32 (s, 3H). 17
Ethyl 5-{[5-chloro-4-({2- [(methylamino)carbonyl] phenyl}amino)-2-
pyridinyl]amino}-1- ethyl-1H-pyrazole-3- carboxylate ##STR00041##
##STR00042## LCMS (ES) m/z = 443.1 (M + H); 1H NMR (400 MHz,
METHANOL-d4) .delta. ppm 8.02 (s, 1H), 7.75-7.74 (m, 1H), 7.62-7.60
(m, 2H), 7.40-7.36 (m, 1H), 6.82 (s, 1H), 6.59 (s, 1H), 4.41-4.36
(m, 2H), 4.19-4.14 (m, 2H), 2.91 (s, 3H), 1.43-1.37 (m, 6H). 18
5-{[5-Chloro-4-({2- [(methylamino)carbonyl] phenyl}amino)-2-
pyridinyl]amino}-1- ethyl-1H-pyrazole-3- carboxylic acid
##STR00043## ##STR00044## LCMS (ES) m/z = 415.1 (M + H); 1H NMR
(400 MHz, METHANOL-d4) .delta. ppm 8.02 (s, 1H), 7.78-7.76 (m, 1H),
7.65-7.63 (m, 2H), 7.44-7.40 (m, 1H), 6.83 (s, 1H), 6.59 (s, 1H),
4.20-4.14 (m, 2H), 2.91 (s, 3H), 1.42 (tr, J = 6.8 Hz, 3H). 19
5-{[5-Chloro-4-({2- [(methylamino)carbonyl] phenyl}amino)-2-
pyridinyl]amino}-1- ethyl-N-(methyloxy)-1H- pyrazole-3-carboxamide
##STR00045## ##STR00046## LCMS (ES) m/z = 444.1 (M + H); 1H NMR
(400 MHz, METHANOL-d4) .delta. ppm 8.01 (s, 1H), 7.76-7.74 (m, 1H),
7.63-7.62 (m, 2H), 7.41-7.37 (m, 1H), 6.74 (s, 1H), 6.57 (s, 1H),
4.16-4.13 (m, 2H), 3.81 (s, 3H), 2.91 (s, 3H), 1.42 (tr, J = 6.8
Hz, 3H). 20 5-{[5-Chloro-4-({2- [(methylamino)carbonyl]
phenyl}amino)-2- pyridinyl]amino}-1- ethyl-N-methyl-1H-
pyrazole-3-carboxamide ##STR00047## ##STR00048## LCMS (ES) m/z =
428.1 (M + H); 1H NMR (400 MHz, METHANOL-d4) .delta. ppm 8.01 (s,
1H), 7.76-7.74 (m, 1H), 7.62-7.61 (m, 2H), 7.41-7.38 (m, 1H), 6.71
(s, 1H), 6.55 (s, 1H), 4.16-4.10 (q, J = 7.2 Hz, 2H), 2.92 (s, 3H),
2.91 (s, 3H), 1.42 (tr, J = 7.2 Hz, 3H). 21 2-[(5-Chloro-2-{[3-
methyl-1-(2,2,2- trifluoroethyl)-1H- pyrazol-5-yl]amino}-4-
pyridinyl)amino]-N- methylbenzamide ##STR00049## ##STR00050## LCMS
(ES) m/z = 439.1 (M + H); 1H NMR (400 MHz, METHANOL-d4) .delta. ppm
8.00 (8, 1H), 7.76-7.74 (m, 1H), 7.62-7.58 (m, 2H), 7.40-7.36 (m,
1H), 6.61 (s, 1H), 6.25 (s, 1H), 4.82-4.76 (q, J = 8.4 Hz, 2H),
2.91 (s, 3H), 2.27 (s, 3H). 22 2-[(5-Chloro-2-{[1-(1-
methyl-4-piperidinyl)- 1H-pyrazol-5-yl]amino}-
4-pyridinyl)amino]-N- methylbenzamide ##STR00051## ##STR00052##
LCMS (ES) m/z = 440.1 (M + H); 1H NMR (400 MHz, METHANOL-d4)
.delta. ppm 8.04 (s, 1H), 7.75-7.73 (m, 1H), 7.61-7.57 (m, 3H),
7.42-7.38 (m, 1H), 6.45 (s, 1H), 6.33 (s, 1H), 4.60-4.50 (m, 1H),
3.67-3.63 (m, 2H), 3.25-3.15 (m, 2H), 2.91 (s, 3H), 2.90 (s, 3H),
2.40-2.12 (m, 4H). 23 2-{[5-Chloro-2-({1-[2-
(dimethylamino)ethyl]-3- methyl-1H-pyrazol-5- yl}amino)-4-
pyridinyl]amino}-N- methylbenzamide ##STR00053## ##STR00054## LCMS
(ES) m/z = 428.1 (M + H); 1H NMR (400 MHz, METHANOL-d4) .delta. ppm
8.06 (s, 1H), 7.76-7.74 (m, 1H), 7.62-7.61 (m, 2H), 7.43-7.38 (m,
1H), 6.59 (s, 1H), 6.18 (s, 1H), 4.41-4.38 (t, J = 5.6 Hz, 2H),
3.63-3.60 (t, J = 5.6 Hz, 2H), 2.98 (s, 6H), 2.91 (s, 3H), 2.25 (s,
3H). 24 5-{[5-Chloro-4-({2- [(methylamino)carbonyl]
phenyl}amino)-2- pyridinyl]amino}-N-[2- (dimethylamino)ethyl]-1-
ethyl-1H-pyrazole-3- carboxamide ##STR00055## ##STR00056## LCMS
(ES) m/z = 485.2 (M + H); 1H NMR (400 MHz, METHANOL-d4) .delta. ppm
8.03 (s, 1H), 7.76-7.74 (m, 1H), 7.62-7.60 (m, 2H), 7.40-7.35 (m,
1H), 6.76 (s, 1H), 6.56 (s, 1H), 4.16-4.14 (q, J = 7.2 Hz, 2H),
3.77 (tr, J = 6.0 Hz, 2H), 3.38 (tr, J = 6.0 Hz, 2H), 2.99 (s, 6H),
2.91 (s, 3H), 1.42 (tr, J = 7.2 Hz, 3H). 25 5-{[5-Chloro-4-({2-
[(methylamino)carbonyl] phenyl}amino)-2- pyridinyl]amino}-N-[2-
(dimethylamino)ethyl]-1- ethyl-N-methyl-1H- pyrazole-3-carboxamide
##STR00057## ##STR00058## LCMS (ES) m/z = 499.2 (M + H); 1H NMR
(400 MHz, METHANOL-d4) .delta. ppm 8.04 (s, 1H), 7.76-7.74 (m, 1H),
7.62-7.60 (m, 2H), 7.40-7.35 (m, 1H), 6.70 (s, 1H), 6.52 (s, 1H),
4.16-4.10 (m, 2H), 3.95-3.90 (m, 1H), 3.58-3.40 (m, 5H), 3.20-3.15
(m, 1H), 3.03 (s, 6H), 2.91 (s, 3H), 1.42 (tr, J = 7.2 Hz, 3H). 26
2-({5-Chloro-2-[(1,3- dimethyl-1H-pyrazol-4- yl)amino]-4-
pyridinyl}amino)-N- methylbenzamide ##STR00059## ##STR00060## LC-MS
(ES) m/z = 371.1 (M + H).sup.+; .sup.1H NMR (CD.sub.3OD, 400 MHz)
.delta. 7.88 (s, 1H), 7.84 (s, 1H), 7.75 (m, 1H), 7.64 (m, 2H),
7.40 (m, 1H), 6.58 (s, 1H), 3.92 (s, 3H), 2.92 (s, 3H), 2.18 (s,
3H) 27 2-({5-Chloro-2-[(1,3- dimethyl-1H-pyrazol-4- yl)amino]-4-
pyridinyl}amino)-N- methylbenzamide ##STR00061## ##STR00062## LC-MS
(ES) m/z = 425.0 (M + H).sup.+, .sup.1H NMR (CD.sub.3OD, 400 MHz)
.delta. 7.98 (s, 1H), 7.90 (s, 1H), 7.76 (m, 1H), 7.63 (m, 2H),
7.40 (m, 1H), 6.58 (s, 1H), 4.00 (s, 3H), 2.91 (s, 3H)
[0167] The
2-{[5-trifluoromethyl-2-(aminopyrazole)-4-pyridinyl]amino}-benz-
amide compounds illustrated in Table 2 were prepared from
2-[(2-chloro-5-trifluoromethyl-4-pyridinyl)amino]-methylbenzamide
and the corresponding amino-pyrazole substantially according to the
procedure of Example 2.
TABLE-US-00003 TABLE 2 ##STR00063## Ex Name Pyr Data 28
2-{[2-[(1,3-Dimethyl-1H-pyrazol- 5-yl)amino]-5-(trifluoromethyl)-4-
pyridinyl]amino}-N- methylbenzamide ##STR00064## LC-MS (ES) m/z =
405.1 (M + H).sup.+ 29 2-{[2-[(1-Ethyl-1H-pyrazol-5-
yl)amino]-5-(trifluoromethyl)-4- pyridinyl]amino}-N-
methylbenzamide ##STR00065## LC-MS (ES) m/z = 405.1 (M + H).sup.+
30 N-Methyl-2-{[2-{[3-methyl-1-(1- methylethyl)-1H-pyrazol-5-
yl]amino}-5-(trifluoromethyl)-4- pyridinyl]amino}benzamide
##STR00066## LC-MS (ES) m/z = 433.2 (M + H).sup.+ 31 2-{[2-({1-[2-
(Dimethylamino)ethyl]-3-methyl- 1H-pyrazol-5-yl}amino)-5-
(trifluoromethyl)-4- pyridinyl]amino}-N- methylbenzamide
##STR00067## LC-MS (ES) m/z = 462.2 (M + H).sup.+ 32
2-{[2-[(1-Ethyl-3-methyl-1H- pyrazol-5-yl)amino]-5-
(trifluoromethyl)-4- pyridinyl]amino}-N- methylbenzamide
##STR00068## LCMS (ES) m/z = 419.2 (M + H) 33
N-Methyl-2-{[2-[(3-methyl-1- phenyl-1H-pyrazol-5-yl)amino]-5-
(trifluoromethyl)-4- pyridinyl]amino}benzamide ##STR00069## LCMS
(ES) m/z = 467.1 (M + H) 34 2-{[2-{[1-(2-hydroxyethyl)-3-
methyl-1H-pyrazol-5-yl]amino}- 5-(trifluoromethyl)-4-
pyridinyl]amino}-N- methylbenzamide ##STR00070## LCMS (ES) m/z =
435.1 (M + H)
Intermediate 1
2-[(2,5-Dichloro-4-pyridinyl)amino]benzoic acid
##STR00071##
[0169] A mixture of 2,5-dichloro-4-iodopyridine (10 g, 36.5 mmol),
2-aminobenzoic acid (4.85 g, 35.4 mmol), DPEPhos
[bis(2-diphenylphosphinophenyl)ether] (1.6 g, 2.97 mmol),
palladium(II) acetate (160 mg, 0.713 mmol) and K.sub.3PO.sub.4 (20
g, 94 mmol) was degassed and heated at 120.degree. C. (oil bath
temp) for 20 h. After 20 h, LCMS showed there was 33% (relative to
the desired product) starting material left. Added another 160 mg
of Pd(OAc).sub.2 to the mixture, and heated to 120.degree. C. for
another 24 h. LCMS showed conversion complete. The mixture was
cooled to room temperature, followed by filtration, and washing
with EtOAc. The solids were acidified to pH=7-8, followed by
filtration. However, the mixture was a paste, and collected solids
could not be dried completely. The solids (11 g) was acidified with
6N HCl to pH=1. The resulting paste was filtered, and washed with
water and TBME. The solid was dried under vacuum over
P.sub.2O.sub.5 for 2 days to give the title compound (7.32 g, 60.2%
yield). MS: M(C.sub.12H.sub.8Cl.sub.2N.sub.2O.sub.2)=283.11,
(M+H).sup.+=283.8; 1H NMR (400 MHz, DMSO) ppm 13.6 (s, 1H) 10.2 (s,
1H) 8.3 (s, 1H) 8.0 (d, 1H) 7.6 (q, 2H) 7.3 (s, 1H) 7.2 (m,
1H).
Example 35
2-({5-Chloro-2-[(1,3-dimethyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amino)ben-
zoic acid
##STR00072##
[0171] A pressure tube was charged with
2-[(2,5-dichloro-4-pyridinyl)amino]benzoic acid (1.0 g, 3.53 mmol),
1,3-dimethyl-1H-pyrazol-5-amine (0.589 g, 5.30 mmol),
(.+-.)-2,2'-Bis(diphenylphosphino)-1,1'-binaphthalene (0.330 g,
0.530 mmol), tris(dibenzylideneacetone)dipalladium(0)
(Pd.sub.2(dba).sub.3, 0.162 g, 0.177 mmol) and sodium tert-butoxide
(0.849 g, 8.83 mmol) in 1,4-dioxane (30 mL). The tube was degassed
with N.sub.2 and sealed and the reaction mixture was heated in an
oil bath at 120.degree. C. for 18 hours. The reaction mixture was
evaporated to dryness under high vacuum. The residue was taken back
in water solution and the pH was adjusted to .about.4 to 5 using
6.0 N hydrochloride acid. The reaction was concentrated to dryness
and the resulting solid was dissolved in MeOH and purified by
reverse-phase HPLC to give the title compound as a solid (285 mg,
21% yield). MS: M(C.sub.17H.sub.16ClN.sub.5O.sub.2)=357.79,
(M+H).sup.+=358, 360.
Example 36
2-({5-Chloro-2-[(1,3-dimethyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amino)-N--
(methyloxy)benzamide
##STR00073##
[0173] A vessel was charged with
3-({5-chloro-2-[(1,3-dimethyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amino)be-
nzoic acid (100 mg, 0.279 mmol),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (53.6
mg, 0.279 mmol) and hydroxybenzotriazole (42.8 mg, 0.279 mmol) in
N,N-dimethylformamide (DMF, 1.0 mL) and the contents were stirred
at room temperature for 30 min. Methoxylamine hydrochloride (23.34
mg, 0.279 mmol) was added to this mixture and stirring continued
for another 10 min. The reaction mixture was cooled to 0.degree. C.
Diisopropylethylamine (DIEA, 0.098 mL, 0.559 mmol) was added and
the reaction mixture was stirred at room temperature overnight. The
final crude material was purified using reverse-phase HPLC,
(Gilson) eluted with CH.sub.3CN/H.sub.2O with 0.1% formic acid to
yield 15 mg (18% yield) MS:
M(C.sub.18H.sub.19ClN.sub.6O.sub.2)=386.84, (M+H).sup.+=387; 1H NMR
(400 MHz, MeOD) ppm 8.16 (s, 1H) 7.93 (s, 1H) 7.50-7.66 (m, 2H)
7.11-7.25 (m, 1H) 6.62 (s, 1H) 5.99 (s, 1H) 3.80 (s, 3H) 3.55-3.70
(s, 3H) 2.11-2.26 (s, 3H).
[0174] The
2-({5-chloro-2-[(1,3-dimethyl-1H-pyrazol-5-yl)amino]-4-pyridiny-
l}amino)-N-alkyl-N-(alkylyloxy)benzamide compounds illustrated in
Table 3 were prepared from
3-({5-chloro-2-[(1,3-dimethyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amino)be-
nzoic acid and amino-alcohols substantially according to the
procedure of Example 36.
TABLE-US-00004 TABLE 3 ##STR00074## Ex Name R.sub.c Data 37
2-({5-chloro-2-[(1,3-dimethyl- 1H-pyrazol-5-yl)amino]-4-
pyridinyl}amino)-N-methyl- N-(methyloxy)benzamide ##STR00075##
LC-MS (ES) m/z = 387 (M + H).sup.+; 1H NMR (400 MHz, MeOD) ppm 8.16
(s, 1 H) 7.93 (s, 1 H) 7.50-7.66 (m, 2H) 7.11- 7.25 (m, 1 H) 6.62
(s, 1 H) 5.99 (s, 1 H) 3.80 (s, 3 H) 3.55-3.70 (s, 3 H) 2.11-2.26
(s, 3H) 38 2-({5-Chloro-2-[(1,3-dimethyl- 1H-pyrazol-5-yl)amino]-4-
pyridinyl}amino)-N-{[2- (dimethylamino)ethyl]oxy} benzamide
##STR00076## LC-MS (ES) m/z = 444, 446 (M + H).sup.+; 1H NMR (400
MHz, MeOD) .delta. ppm 8.50 (s, 1 H) 7.93 (s, 1 H) 7.64-7.71 (m, 1
H) 7.51- 7.61 (m, 2H) 7.21 (ddd, J = 8.0, 6.2, 2.3 Hz, 1 H) 6.58
(s, 1 H) 5.98 (s, 1 H) 4.22-4.31 (m, 2H) 3.61 (s, 3 H) 3.33--3.40
(m, 2H) 2.96 (s, 6 H) 2.19 (s, 3 H)
Example 39
2-({5-Chloro-2-[(1-ethyl-3-methyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amino-
)-N-(methyloxy)benzamide
##STR00077##
[0175] 39a)
2-[(2,5-Dichloro-4-pyridinyl)amino]-N-(methyloxy)benzamide
##STR00078##
[0177] A vessel was charged with
3-[(2,5-dichloro-4-pyridinyl)amino]benzoic acid (1.0 g, 3.53 mmol),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (677
mg, 3.53 mmol) and hydroxybenzotriazole (HOBT) (541 mg, 3.53 mmol)
in N,N-dimethylformamide (DMF, 7.0 mL) and was stirred at room
temperature for 30 min. To this solution methoxylamine
hydrochloride (0.3 g, 3.53 mmol) was added and reaction mixture was
stirred for another 10 min. The reaction mixture was cooled to
0.degree. C. by using an ice bath. To this reaction mixture
diisopropylethylamine (1.2 mL, 7.06 mmol) was added and the mixture
was stirred at room temperature overnight. After concentrating
under vacuum, the residue was worked up by using a saturated
aqueous solution of NaHCO.sub.3 and CH.sub.2Cl.sub.2. The organic
phase was washed with brine then dried over MgSO.sub.4 and
filtered. The CH.sub.2Cl.sub.2 was removed by rotary evaporation.
The crude material was loaded on silica gel column and eluted by
MeOH in CH.sub.2Cl.sub.2 with NH.sub.4OH 0.1%, which gave the
desired product
2-[(2,5-dichloro-4-pyridinyl)amino]-N-(methyloxy)benzamide (850 mg,
2.72 mmol, 77% yield) MS:
M(C.sub.13H.sub.11C.sub.12N.sub.3O.sub.2)=312.15, (M+H).sup.+=312,
314; 1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 9.57 (br. s., 1H)
8.72 (s, 1H) 8.22 (s, 1H) 7.51-7.67 (m, 3H) 7.25 (s, 1H) 7.07-7.21
(m, 1H) 3.92 (s, 3H).
39b)
2-({5-Chloro-2-[(1-ethyl-3-methyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}-
amino)-N-(methyloxy)benzamide
[0178] A 20-mL microwave tube was charged with
2-[(2,5-dichloro-4-pyridinyl)amino]-N-(methyloxy)benzamide (100 mg,
0.320 mmol), 1-ethyl-3-methyl-1H-pyrazol-5-amine (60.1 mg, 0.481
mmol), cesium carbonate (313 mg, 0.961 mmol), 1,4-dioxane (5.0 mL)
and THF (1.0 mL). The reaction mixture was degassed by nitrogen for
10 min. (.+-.)-2,2'-Bis(diphenylphosphino)-1,1'-binaphthalene
(19.95 mg, 0.032 mmol) and Palladium(II) acetate (3.60 mg, 0.016
mmol) in minimum amount of 1,4-dioxane were then added. The tube
was sealed and reaction mixture was heated in microwave oven
160.degree. C. for 40 min. The resulting suspension was cooled to
room temperature and filtered through celite. The filtrate was
evaporated to dryness and the crude reaction mixture was purified
by reverse-phase HPLC to give the title compound as a solid (16 mg,
24% yield) MS: M(C.sub.19H.sub.21ClN.sub.6O.sub.2)=400.86,
(M+H).sup.+=401; 1H NMR (400 MHz, MeOD) .delta. ppm 7.92 (s, 1H)
7.44-7.66 (m, 3H) 7.11-7.25 (m, 1H) 6.61 (s, 1H) 5.99 (s, 1H) 3.98
(q, J=7.3 Hz, 2H) 3.80 (s, 3H) 2.21 (s, 3H) 1.32 (t, J=7.2 Hz,
3H).
[0179] The
2-({5-chloro-2-(pyrazol-5-yl)amino-]-4-pyridinyl}amino)-N-(meth-
yloxy)benzamide compounds illustrated in Table 4 were prepared from
2-[(2,5-dichloro-4-pyridinyl)amino]-N-(methyloxy)benzamide and
amino-pyrazoles substantially according to the procedure of Example
39.
TABLE-US-00005 TABLE 4 ##STR00079## Ex Name Pyr Data 40
2-({5-Chloro-2-[(1-ethyl- 1H-pyrazol-5-yl)amino]-4-
pyridinyl}amino)-N- (methyloxy)benzamide ##STR00080## LC-MS (ES)
m/z = 387, 388 (M + H).sup.+; 1H NMR (400 MHz, MeOD) .delta. ppm 1H
7.90-7.96 (m, 1 H) 7.49-7.66 (m, 3 H) 7.45 (t, J = 2.2 Hz, 1 H)
7.10- 7.22 (m, 1 H) 6.61 (s, 1 H) 6.19 (d, J = 1.8 Hz, 1 H) 4.06
(q, J = 7.1 Hz, 2H) 3.80 (s, 3 H) 1.35 (t, J = 7.2 Hz, 3H)
Intermediate 2
2-[(2,5-Dichloro-4-pyridinyl)amino]-N-(methyloxy)benzamide
##STR00081##
[0181] A solution of 3-[(2,5-dichloro-4-pyridinyl)amino]benzoic
acid (500 mg, 1.766 mmol),
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (EDC) (339 mg, 1.766
mmol) and 1-hydroxybenzotriazole (HOBT) (270 mg, 1.766 mmol) in
N,N-dimethylformamide (3532 .mu.l) was stirred at room temperature
for 30 min. To this solution O-methylhydroxylamine(aminoxy)methane
(148 mg, 1.766 mmol) was then added and stirred for another 10 min.
The reaction mixture was cooled using an ice water bath. Then
diisopropylethylamine (617 .mu.l, 3.53 mmol) was added. After the
addition was finished, the reaction mixture was stirred at room
temperature overnight. The reaction mixture was followed by HPLC
and LCMS. The final crude material was worked up by addition of
saturated aqueous NaHCO.sub.3 and CH.sub.2Cl.sub.2. Organic phase
was washed with brine then dried over MgSO.sub.4. The solution was
filtered and solvent was removed by evaporation. The oil like crude
material was loaded on silica column and eluted with MeOH in
CH.sub.2Cl.sub.2 with NH.sub.4OH 0.1% to give the target compound
2-[(2,5-dichloro-4-pyridinyl)amino]-N-(methyloxy)benzamide (320 mg,
1.025 mmol, 58.0% yield) as a yellow solid; MS;
M(C.sub.13H.sub.11Cl.sub.2N.sub.3O.sub.2)=312.15, (M+H).sup.+=312,
313.9; 1H NMR (400 MHz, CHLOROFORM-d) ppm 9.66 (br. s., 1H) 9.60
(br. s., 1H) 8.20 (s, 1H) 7.49-7.61 (m, 3H) 7.24 (s, 1H) 7.09-7.16
(m, 1H) 3.90 (s, 3H).
Example 41a
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyrid-
inyl)amino]-N-(methyloxy)benzamide
##STR00082##
[0183] A microwave tube was charged with
2-[(2,5-dichloro-4-pyridinyl)amino]-N-(methyloxy)benzamide (70 mg,
0.224 mmol), {3-methyl-1-(1-methylethyl)-1H-pyrazol-5-amine (70 mg,
0.503 mmol) and cesium carbonate (230 mg, 0.706 mmol). The reaction
mixture was degassed with nitrogen for 10 min. At same time, BINAP
(50 mg, 0.080 mmol) and palladium(II) acetate (10 mg, 0.045 mmol)
were added. The reaction mixture was heated in a microwave at
160.degree. C. for 40 min. The crude material was purified on
reverse-phase HPLC (Gilson) eluting with CH.sub.3CN/H.sub.2O with
0.1% formic acid which gave a title compound (15 mg, 15%); MS:
M(C.sub.20H.sub.23ClN.sub.6O.sub.2)=414.89, (M+H).sup.+=415, 416;
1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 9.42 (br. s., 1H) 8.71
(br. s., 1H) 8.02 (s, 1H) 7.54 (br. s., 1H) 7.06 (t, J=7.5 Hz, 1H)
6.48 (s, 1H) 6.32 (br. s., 1H) 5.86 (s, 1H) 4.47 (dt, J=13.4, 6.7
Hz, 1H) 3.92 (s, 3H) 2.26 (s, 3H) 1.41-1.43 (d, J=6.6 Hz, 2H).
Intermediate 3
2-[(2,5-Dichloro-4-pyridinyl)amino]benzonitrile
##STR00083##
[0185] The solution of 2,5-dichloro-4-iodopyridine (100 g, 365
mmol), 2-aminobenzonitrile (43.1 g, 365 mmol) and potassium
triphosphate (233 g, 1095 mmol) in 1,4-dioxane (2.5 L) was degassed
by N.sub.2 stream. To this solution was added DPEPhos (15.73 g,
29.2 mmol) and palladium acetate (3.28 g, 14.60 mmol). The reaction
mixture was stirred at reflux for 18 hour. The solution was
filtered through 0.5 in. celite and 0.2 inch of silica. The
solution was evaporated. Solid was suspended in the diethyl ether
and filtered. Diethyl ether was concentrated, and the resulting
solid was filtered. 2-[(2,5-Dichloro-4-pyridinyl)amino]benzonitrile
(80 g, 288 mmol, 79% yield) was isolated as an orange solid. 1H NMR
(400 MHz, DMSO-d.sub.6) ppm 6.49 (s, 1H) 7.50 (td, J=7.58, 1.01 Hz,
1H) 7.56 (d, J=7.58 Hz, 1H) 7.80 (td, J=7.83, 1.77 Hz, 1H) 7.95
(dd, J=7.83, 1.52 Hz, 1H) 8.26 (s, 1H) 9.05 (brs, 1H); HPLC Rt=2.88
min, MS (ESI): 263.9, 265.9 [M+H].sup.+.
Intermediate 4
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyrid-
inyl)amino]benzonitrile
##STR00084##
[0187] The solution of
2-[(2,5-dichloro-4-pyridinyl)amino]benzonitrile (110 g, 396 mmol),
3-methyl-1-(1-methylethyl)-1H-pyrazol-5-amine (55.1 g, 396 mmol),
and cesium carbonate (387 g, 1187 mmol) in 1,4-dioxane (2.5 L) was
degassed by N.sub.2 stream, and
2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (BINAP) (19.71 g, 31.7
mmol) followed by palladium acetate (3.55 g, 15.83 mmol) were
added. The reaction mixture was heated to reflux for overnight
under N.sub.2. The reaction mixture was filtered and the liquid was
concentrated. Ethyl acetate (1500 mL), followed by 1 M HCl (1000
mL) were added. Layers were separated. Ethyl acetate was washed
with 1 M HCl until no product was observed by HPLC (1000 mL total,
1.times.). HCl phases were combined, and backwashed with ethyl
acetate (3.times.1000 mL), until the product peak was relativity
pure in the HCL layer. The HCl layer was then basified with NaOH
(50 w/w followed by 1 M) to ph .about.4 resulting in a cloudy
solution. Ethyl acetate (2000 mL) was added and layers were
separated. The ethyl acetate was washed with brine and evaporated.
After neutralization--after addition of ethyl acetate--the reaction
mixture was filtered to get some product. Also isolation of product
during evaporation can be done by filtration of white solid, which
comes from the mother liquor. All solids and evaporated products
were combined.
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]benzonitrile (80 g, 207 mmol, 52.4% yield) was isolated
as a yellow solid. 1H NMR (400 MHz, DMSO-d.sub.6) ppm 1.24 (d,
J=6.57 Hz, 6H) 2.08 (s, 3H) 4.34 (quin, J=6.57 Hz, 1H) 5.87 (s, 1H)
5.97 (s, 1H) 7.41 (td, J=7.58, 1.01 Hz, 1H) 7.47 (d, J=8.08 Hz, 1H)
7.75 (td, J=7.83, 1.52 Hz, 1H) 7.90 (dd, J=7.83, 1.52 Hz, 1H) 7.94
(s, 1H) 8.42 (d, J=17.43 Hz, 2H); HPLC Rt=2.36 min, MS (ESI):
[M+H].sup.+=367.1, 368.1.
Intermediate 5
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyrid-
inyl)amino]benzoic acid
##STR00085##
[0189]
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}--
4-pyridinyl)amino]benzonitrile (80 g, 218 mmol) was dissolved in
1,4-dioxane (1.5 L) and 1 M NaOH (1500 mL, 1500 mmol) was added.
The suspension was refluxed overnight. After cooling to RT, ethyl
acetate (1 L) was added and layers were separated. The water layer
was washed with 1 L of ethyl acetate. Both organic layers were
combined and backwashed with 0.1 M NaOH (1 L) until no product was
observed in organic. The organics were then discarded. Combined
aqueous were then washed with 1 L of ethyl acetate. The water layer
was then acidified with acetic acid (very slowly to ph .about.7).
The solid was filtered and
2-[(5-chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]benzoic acid (67 g, 165 mmol, 76% yield) was isolated
as a yellow solid. 1H NMR (400 MHz, DMSO-d.sub.6) ppm 1.28 (d,
J=6.57 Hz, 6H) 2.11 (s, 3H) 4.41 (quin, J=6.57 Hz, 1H) 5.96 (s, 1H)
6.83 (s, 1H) 7.09 (ddd, J=8.02, 5.12, 3.03 Hz, 1H) 7.40 (1H)
7.52-7.61 (m, 2H) 7.91-8.16 (m, 2H) 8.55 (s, 1H) 10.17 (brs, 1H)
13.64 (brs, 1H); HPLC Rt=2.35 min, MS (ESI): [M+H].sup.+=386.1.
Example 41b
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyrid-
inyl)amino]-N-(methyloxy)benzamide
##STR00086##
[0191] To a solution of
2-[(5-chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]benzoic acid (67 g, 174 mmol) and
1-hydroxybenzotriazole (29.3 g, 191 mmol) in N,N-dimethylformamide
(700 mL) was added N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide
(36.6 g, 191 mmol) and the solution was stirred for 30 minutes.
O-Methylhydroxylamine hydrochloride (15.95 g, 191 mmol) was added
and the solution stirred for additional 15 minutes, the cooled down
to the 0.degree. C. and diisopropylethlyamine (91 mL, 521 mmol) was
added dropwise. The reaction mixture was stirred overnight at the
room temperature. Water (4000 mL) was added and the solution was
acidified with acetic acid (20 mL). The solution was extracted
2.times.2 L of ethyl acetate. The organic was washed with water (1
L), brine, and dried over MgSO.sub.4, filtered and evaporated.
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-N-(methyloxy)benzamide (74 g, 164 mmol, 94% yield, 92%
pure) was isolated as a yellow foam. 1H NMR (400 MHz, DMSO-d.sub.6)
ppm 1.27 (d, J=6.57 Hz, 6H) 2.10 (s, 3H) 3.71 (s, 3H) 4.39 (quin,
J=6.51 Hz, 1H) 5.93 (s, 1H) 6.66 (s, 1H) 7.08-7.19 (m, 1H)
7.49-7.64 (m, 3H) 7.98 (s, 1H) 8.50 (s, 1H) 9.50 (s, 1H) 11.93 (s,
1H); HPLC Rt=2.13 min, MS (ESI): [M+H].sup.+=415.1.
Purification of Example 41a and 41b Products
##STR00087##
[0193]
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}--
4-pyridinyl)amino]-N-(methyloxy)benzamide (173.3 g, 63.5% w/w,
265.2 mmoles) was dissolved in ethyl acetate (3.50 L, 20 volumes)
and heated to about 50.degree. C. To this solution was added
Si-thiol (functionalized silica gel) (87 g, 50% loading). The
mixture was held at about 50.degree. C. for 16-20 hours. It was
then filtered off the Si-thiol silica gel. The filter cake was
rinsed with ethyl acetate (2.times.200 mL each) and filtrates were
combined. Then the combined filtrates were washed with 1 M aqueous
ammonium formate at pH 9.4 (5.times.1 L each), washed with water,
brine, and dried over magnesium sulfate. Dried EtOAC was filtered
and stripped to dryness giving a yellow foam. It was dried at
50-55.degree. C. for about 2 hours to a constant weight of 160 g.
This material was slurried in methylene chloride (800 mL, 5
volumes), heated to reflux to afford a solution, and filtered. The
solution was cooled to 20-25.degree. C. The product crystallized
upon cooling. After about 2 hours, the product was collected by
filtration and rinsed with methylene chloride. The white solid was
dried at 50-55.degree. C. for 14-16 hours to a constant weight.
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-N-(methyloxy)benzamide (85.0 g, 204.9 mmoles, 77%
overall yield) was isolated as a white solid. 1H NMR (400 MHz,
DMSO-d.sub.6) ppm 1.27 (d, J=6.57 Hz, 6H) 2.10 (s, 3H) 3.70 (s, 3H)
4.39 (quin, J=6.57 Hz, 1H) 5.92 (s, 1H) 6.66 (s, 1H) 7.02-7.24 (m,
1H) 7.45-7.68 (m, 3H) 7.98 (s, 1H) 8.48 (s, 1H) 9.49 (br. s, 1H)
11.91 (s, 1H). C18 HPLC RT=6.2 minutes (99.0% purity). MS (ESI):
415.0 [M+H].sup.+.
##STR00088##
[0194]
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}--
4-pyridinyl)amino]-N-(methyloxy)benzamide (235.2 g total weight,
228.0 g assayed content, 549.5 mmoles) was slurried in ethyl
acetate (7.1 L, 30 volumes). The mixture was heated to about
50-55.degree. C. to afford a cloudy solution. The cloudy solution
was filtered. To the filtered solution was added 2.0 M HCl in
diethyl ether (210 g, 281 mL, 1.02 equiv.) over 15-20 minutes. Upon
HCl addition, a white slurry was observed. It was stirred at room
temperature for about 16-20 hours. Product was collected by
filtration and rinsed with ethyl acetate (2.times.500 mL each). The
wet cake was dried at 50-55.degree. C./<5 mm Hg for 16-20 hours
to a constant weight.
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-N-(methyloxy)benzamide, monohydrochloride, (245.9 g,
544.7 mmoles, 96% yield) was isolated as a white solid. 1H NMR (400
MHz, DMSO-d.sub.6) ppm 1.32 (d, J=6.57 Hz, 6H) 2.18 (s, 3H) 3.70
(s, 3H) 4.35-4.62 (m, 1H) 6.12 (br. s, 1H) 6.60 (br. s, 1H)
7.19-7.41 (m, 1H) 7.48-7.75 (m, 3H) 8.09 (s, 1H) 9.59-9.99 (m, 2H)
11.98 (br. s, 1H). C18 HPLC RT=6.1 minutes (99.6% purity). MS
(ESI): 414.8 [M+H].sup.+.
Example 42
2-({5-Chloro-2-[(1-ethyl-3-methyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amino-
)-N-ethylbenzamide
##STR00089##
[0196] A vessel was charged with
2-[(2,5-dichloro-4-pyridinyl)amino]-N-ethylbenzamide (100 mg, 0.322
mmol), 1-ethyl-3-methyl-1H-pyrazol-5-amine (60.5 mg, 0.484 mmol),
cesium carbonate (315 mg, 0.967 mmol), 1,4-dioxane (5.0 mL), and
THF (1.0 mL). The reaction mixture was degassed by nitrogen for 10
min at which time
(.+-.)-2,2'-bis(diphenylphosphino)-1,1'-binaphthalene (40.1 mg,
0.064 mmol) and palladium (II) acetate (7.24 mg, 0.032 mmol) in a
minimum amount of 1,4-dioxane were added. The vessel was sealed and
the reaction mixture was heated in microwave oven 160.degree. C.
for 40 min. The resulting suspension was cooled to room temperature
and filtered through celite. The filtrate was evaporated to dryness
and the crude reaction mixture was purified by reverse-phase HPLC
to give the title compound as a solid (45 mg, 30% yield); MS:
M(C.sub.20H.sub.23ClN.sub.6O)=398.89, (M+H).sup.+=399, 401; 1H NMR
(400 MHz, CHLOROFORM-d) .delta. ppm 8.34 (s, 1H) 7.94 (s, 1H) 7.66
(br. s., 1H) 7.51 (d, J=7.8 Hz, 1H) 7.31-7.47 (m, 2H) 6.96-7.17 (m,
1H) 6.58 (s, 1H) 6.19 (t, J=5.2 Hz, 1H) 5.86 (s, 1H) 4.01 (q, J=7.3
Hz, 2H) 3.42-3.62 (m, 2H) 2.24 (s, 3H) 1.37 (t, J=7.3 Hz, 3H) 1.26
(t, J=7.3 Hz, 3H).
[0197] The
2-({5-chloro-2-(1H-pyrazol-5-yl)amino]-4-pyridinyl}amino)-N-eth-
ylbenzamide compounds illustrated in Table 5 were prepared from
2-[(2,5-dichloro-4-pyridinyl)amino]-N-ethylbenzamide and
amino-pyrazoles substantially according to the procedure of Example
42.
TABLE-US-00006 TABLE 5 ##STR00090## Ex Name Pyr Data 43
2-({5-Chloro-2-[(1,3- dimethyl-1H-pyrazol-5- yl)amino]-4-
pyridinyl}amino)-N- ethylbenzamide ##STR00091## LC-MS (ES) m/z =
385 (M + H).sup.+; 1H NMR (400 MHz, MeOD) .delta. ppm 7.91 (s, 1 H)
7.67 (dd, J = 7.8, 1.52 Hz, 1 H) 7.44- 7.58 (m, 2H) 7.14-7.24 (m, 1
H) 6.61 (s, 1 H) 5.98 (s, 1 H) 3.62 (s, 3 H) 3.37- 3.46 (m, 2H)
2.19 (s, 3H) 1.21 (t, J = 7.3 Hz, 3H) 44 2-({5-Chloro-2-[(1-ethyl-
1H-pyrazol-5-yl)amino]-4- pyridinyl}amino)-N- ethylbenzamide
##STR00092## LC-MS (ES) m/z = 415, 416 (M + H).sup.+; 1H NMR (400
MHz, CHLOROFORM- d) .delta. ppm 9.85 (s, 1 H) 8.00 (s, 1 H) 7.46-
7.54 (m, 2H) 7.33-7.44 (m, 2H) 6.88- 7.12 (m, 1 H) 6.72 (br. s., 1
H) 6.52 (s, 1 H) 5.93-6.17 (m, 2H) 4.09 (q, J = 7.2 Hz, 2H)
3.38-3.57 (m, 2H) 1.34-1.49 (m, 3 H) 1.26 (t, J = 7.3 Hz, 3 H) 45
2-[(5-Chloro-2-{[3-methyl- 1-(1-methylethyl)-1H-
pyrazol-5-yl]amino}-4- pyridinyl)amino]-N- ethylbenzamide
##STR00093## LC-MS (ES) m/z = 413 (M + H).sup.+; 1H NMR (400 MHz,
DMSO-d6) .delta. ppm 10.04 (s, 1 H) 8.71 (t, J = 5.4 Hz, 1 H) 8.48
(s, 1 H) 7.97 (s, 1 H) 7.71 (dd, J = 7.8, 1.26 Hz, 1 H) 7.44-7.59
(m, 2H) 7.04-7.19 (m, 1 H) 6.69 (s, 1 H) 5.93 (s, 1 H) 4.30-4.45
(m, 1H) 3.22-3.31 (m, 2H) 2.10 (s, 3 H) 1.29-1.36 (d, J = 8.0 Hz 6
H) 1.12 (t, J = 7.2 Hz, 3H)
Intermediate 6
1-Ethyl-3-[2-(1-pyrrolidinyl)ethyl]-1H-pyrazol-5-amine
##STR00094##
[0198] Int. 6a)
N-{1-Ethyl-3-[2-oxo-2-(1-pyrrolidinyl)ethyl]-1H-pyrazol-5-yl}-2,2,2-trifl-
uoroacetamide
##STR00095##
[0200] Pentafluorophenyl trifluoroacetate (497 mg, 1.773 mmol) was
added dropwise to a stirred solution of
(5-amino-1-ethyl-1H-pyrazol-3-yl) acetic acid (150 mg, 0.887 mmol)
and pyridine (0.143 mL, 1.773 mmol) in DMF (3 mL). The reaction
mixture was stirred for 15 min and pyrrolidine (0.220 mL, 2.66
mmol) was added. The reaction mixture was stirred at 65.degree. C.
for 40 min. The mixture was cooled and quenched with water (5 mL)
and extracted with EtOAc (3.times.). The extract was dried over
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
purified using RP-HPLC to give product (125 mg). MS:
(M+H).sup.+=318.8. .sup.1H NMR (400 MHz, CHLOROFORM-d) ppm 1.32 (t,
J=7.2 Hz, 3H), 1.87-2.06 (m, 4H), 3.46 (t, J=6.8 Hz, 2H), 3.53 (t,
J=6.8 Hz, 2H), 3.66 (s, 2H), 4.07 (q, J=7.2 Hz, 2H), 5.30 (s,
1H).
Int. 6b)
1-ethyl-3-[2-oxo-2-(1-pyrrolidinyl)ethyl]-1H-pyrazol-5-amine
##STR00096##
[0202] To a solution of
N-{1-ethyl-3-[2-oxo-2-(1-pyrrolidinyl)ethyl]-1H-pyrazol-5-yl}-2,2,2-trifl-
uoroacetamide (120 mg, 0.377 mmol) in methanol (1.5 mL) was added
2M HCl (1 mL, 0.377 mmol), and the reaction mixture was stirred at
50.degree. C. for 2 h and concentrated. The residue was neutralized
using saturated NaHCO.sub.3 solution and concentrated. The residue
was dried under high vacuum to give 79 mg and used for next
reaction without further purification. MS: (M+H).sup.+=222.8.
.sup.1H NMR (400 MHz, DMSO-d6) ppm 1.18 (t, J=7.2 Hz, 3H),
1.70-1.85 (m, 4H), 3.24 (t, J=6.8 Hz, 2H), 3.31 (s, 2H), 3.43 (t,
J=6.8 Hz, 2H), 3.78 (q, J=7.2 Hz, 2H), 5.11 (s, 1H).
Int. 6c) 1-ethyl-3-[2-(1-pyrrolidinyl)ethyl]-1H-pyrazol-5-amine
##STR00097##
[0204] To a solution of
1-ethyl-3-[2-oxo-2-(1-pyrrolidinyl)ethyl]-1H-pyrazol-5-amine (600
mg, 1.784 mmol) in tetrahydrofuran (8 mL) cooled with water-ice
bath was added a solution of 2M LAH (1.0 mL, 2.00 mmol) solution
dropwise, and the reaction mixture was stirred for 5 h at rt and 30
min at 50 degree. The reaction mixture was carefully quenched with
methanol followed by water and concentrated. The residue was washed
with DCM/methanol 5 times. The extract was concentrated and the
residue was purified using HPLC under the basic conditions to give
220 mg of product. MS: (M+H).sup.+=208.7. .sup.1H NMR (400 MHz,
CHLOROFORM-d) ppm 1.37 (t, J=7.2 Hz, 3H), 1.79 (m, 4H), 2.56 (m,
4H), 2.71 (m, 2H), 3.42 (m, 2H), 3.94 (q, J=7.2 Hz, 2H), 5.40 (s,
1H).
Example 46
2-{[5-Chloro-2-({1-ethyl-3-[2-(1-pyrrolidinyl)ethyl]-1H-pyrazol-5-yl}amino-
)-4-pyridinyl]amino}-N-methylbenzamide
##STR00098##
[0206] To a 5-mL microwave tube were added
2-[(2,5-dichloro-4-pyridinyl)amino]-N-methylbenzamide (100 mg,
0.338 mmol), 1-ethyl-3-[2-(1-pyrrolidinyl)ethyl]-1H-pyrazol-5-amine
(70.3 mg, 0.338 mmol), cesium carbonate (330 mg, 1.013 mmol), and
1,4-dioxane (2 mL), and the mixture was degassed by bubbling
nitrogen through for 15 min. Palladium (II) acetate (3.79 mg, 0.017
mmol) and BINAP (21.03 mg, 0.034 mmol) were added, and the reaction
mixture was heated at 170.degree. C. with stirring under microwave
conditions for 40 min. The reaction mixture was filtered and
concentrated. The residue was purified by using RP-HPLC under basic
conditions (Gemini 5u C18(2) 110A, AXI. 50.times.30.00 mm 5 micron:
7.3-minute run, 47 mL/min, 40% ACN/H.sub.2O, 0.1% NH.sub.4OH to 90%
ACN/H.sub.2O, 0.1% NH.sub.4OH with UV detection at 254 nm) to give
the title compound (62 mg). MS: (M+H).sup.+=468.1. .sup.1H NMR (400
MHz, DMSO-d.sub.6) ppm 1.21 (t, J=7.2 Hz, 3H), 1.67 (m, 4H), 2.45
(m, 4), 2.60 (m, 4H), 2.77 (d, J=4.0 Hz, 3H), 3.89 (q, J=7.2 Hz,
2H), 6.03 (s, 1H), 6.74 (s, 1H), 7.11 (m, 1H), 7.49 (m, 1H), 7.54
(m, 1H), 7.71 (d, J=7.2 Hz, 1H), 8.00 (s, 1H).
Intermediate 7
2-Amino-N-methoxy-benzamide
##STR00099##
[0208] To a mixture of isatoic anhydride (40 g, 245.39 mmol, 1 eq)
and o-methyl hyroxylamine hydrochloride (30.55 g, 368.09 mmole, 1.5
eq) in EtOH:H.sub.2O (9:1) (1000 mL) was added triethylamine (51.2
mL, 368.09 mmole, 1.5 eq) and resulting mixture was reflux for 4 h.
After completion of reaction solvent was removed under reduced
pressure and residue was diluted with water (500 mL), extracted
with ethyl acetate (3.times.250 mL). Combined organic layer was
dried over sodium sulfate, filtered and concentrated under reduced
pressure. Solid compound so obtained was purified by washing with
diethyl ether and hexane to give the title compound as brown solid
(20 g, 49%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 3.67 (s,
3H), 6.20-6.40 (brs, 2H), 6.44-6.53 (m, 1H), 6.70 (d, 1H, J=7.76
Hz), 7.10-7.19 (m, 1H), 7.30 (d, 1H, J=7.6 Hz), 11.40 (s, 1H).
LC-MS [M+H].sup.+=167.2.
Intermediate 8
2-(2,5-Dichloropyridin-4-ylamino)-N-methoxy-benzamide
(CR637-KS210635-027A1)
##STR00100##
[0210] A mixture of 2,5-dichloro-4-iodo-pyridine (40 g, 146.5
mmole, 1 eq), 2-amino-N-methoxy-benzamide (24.32 g, 146.5 mmole, 1
eq) and K.sub.3PO.sub.4 (77.72 g 366.2 mmole, 2.5 eq) in
1,4-dioxane (600 mL) was degassed with N.sub.2 for 1 h. To this
were added Pd(OAc).sub.2 (0.657 g, 2.93 mmole, 0.02 eq), DPEPhos
(6.31 g, 11.7 mmole, 0.08 eq) and again degassed for 15 min with
N.sub.2. The resulting mixture was stirred at 110.degree. C. for
overnight. After completion of reaction, solid material was
collected by filtration, dissolved in water (500 mL) and extracted
with ethyl acetate (5.times.200 mL). Combined organic layer was
dried over sodium sulfate, filtered and concentrated under reduced
pressure. Solid compound so obtained was purified by washing with
hexane to give the title compound as yellowish solid (40 g, 53%).
.sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 3.63 (s, 3H), 7.06 (s,
1H), 7.15-7.22 (m, 1H), 7.48-7.57 (m, 2H), 7.66-7.67 (d, 1H, J=7.48
Hz), 8.25 (s, 1H), 10.66-11.45 (brs, 1H). LC-MS
[M+H].sup.+=312.3.
Example 47
2-({5-Chloro-2-[(1,5-dimethyl-1H-pyrazol-4-yl)amino]-4-pyridinyl}amino)-N--
(methyloxy)benzamide
##STR00101##
[0212] A microwave tube was charged with
2-[(2,5-dichloro-4-pyridinyl)amino]-N-(methyloxy)benzamide (200 mg,
0.64 mmol), 1,5-dimethyl-1H-pyrazol-4-amine (142 mg, 1.28 mmol),
cesium carbonate (626 mg, 1.92 mmol) and dioxane/THF (3:1 ml). The
reaction mixture was degassed under nitrogen for 10 min and
palladium (II) acetate (5.8 mg, 0.03 mmol) and BINAP (40 mg, 0.06
mmol) were added. The tube was sealed and the mixture was stirred
in an oil bath at 150.degree. C. overnight. The dark brown solution
was filtered thru celite and evaporated. It was dissolved in MeOH
and filtered thru an Acrodisc (Pall Corporation; www.pall.com) and
further purified on preparative Agilent HPLC (5 to 95%
water:acetonitrile with 0.1% formic acid). The dark brown oil
residue was dissolved in DMF and water was slowly added. A tan
precipitate crashed out and was filtered off and dried under vacuum
at 40.degree. C. for 2 hrs to afford the desired product (18 mg,
7.3%) as a tan solid. LC-MS [M+H].sup.+=387.1, 389.1. 1H NMR (400
MHz, DMSO-d.sub.6) .delta. ppm 11.91 (br. s., 1H) 9.41 (br. s., 1H)
7.85-7.98 (m, 2H) 7.49-7.59 (m, 3H) 7.42 (br. s., 1H) 7.07-7.13 (m,
1H) 6.50 (br. s., 1H) 3.69 (d, J=10.36 Hz, 6H) 2.11 (s, 3H).
Example 48
2-({5-Chloro-2-[(1,3-dimethyl-1H-pyrazol-4-yl)amino]-4-pyridinyl}amino)-N--
(methyloxy)benzamide
##STR00102##
[0214] A microwave tube was charged with
2-[(2,5-dichloro-4-pyridinyl)amino]-N-(methyloxy)benzamide (250 mg,
0.8 mmol), 1,3-dimethyl-1H-pyrazol-4-amine (187 mg, 1.68 mmol),
cesium carbonate (783 mg, 2.4 mmol) and dioxane/THF (3:1 ml). The
reaction mixture was degassed under nitrogen for 10 min and
palladium (II) acetate (9 mg, 0.04 mmol) and BINAP (50 mg, 0.08
mmol) were added. The mixture was stirred in a microwave at
140.degree. C. for 40 min. It was evaporated and the residue
dissolved in MeOH was filtered thru celite and thru an Acrodisc and
purified further using preparative Agilent HPLC (5 to 95%
water:acetonitrile with 0.1% formic acid). Fractions were combined
and evaporated. Ether was added to the residue and a tan
precipitate crashed out. It was filtered off and dried under vacuum
at 40.degree. C. for 2 days to afford the desired product (55 mg,
18%) as a tan solid. LC-MS (ES) m/z=387.1, [M+H].sup.+=389.1. 1H
NMR (400 MHz, DMSO-d.sub.6) ppm 11.93 (br. s., 1H) 9.51 (br. s.,
1H) 8.03 (s, 1H) 7.96 (s, 1H) 7.82 (s, 1H) 7.51-7.61 (m, 3H)
7.07-7.15 (m, 1H) 6.68 (s, 1H) 3.70 (d, J=4.29 Hz, 6H) 2.07 (s,
3H).
Example 49
2-[(5-Chloro-2-{[4-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyrid-
inyl)amino]-N-(methyloxy)benzamide
##STR00103##
[0216] A microwave tube was charged with
2-[(2,5-dichloro-4-pyridinyl)amino]-N-(methyloxy)benzamide (100 mg,
0.32 mmol), 4-methyl-1-(1-methylethyl)-1H-pyrazol-5-amine (93.6 mg,
0.67 mmol), cesium carbonate (312.8 mg, 0.96 mmol) and DMF (5 mL).
The reaction mixture was degassed under nitrogen for 10 min and
palladium (II) acetate (3.6 mg, 0.016 mmol) and BINAP (19.9 mg,
0.032 mmol) were added. The reaction mixture was heated in an oil
bath for 6 hours and then in a microwave at 150.degree. C. for 40
min. The solvent was evaporated and the residue dissolved in MeOH.
It was filtered thru celite and thru an Acrodisc to be purified on
preparative Agilent HPLC (5 to 95% water:acetonitrile with 0.1%
formic acid). Fractions were combined and evaporated. The brown oil
residue was diluted in DMF and water was added. A precipitate
crashed out. It was filtered and dried under vacuum at 40.degree.
C. for 6 hrs. LC-MS [M+H].sup.+=415.1. 1H NMR (400 MHz,
DMSO-d.sub.6) ppm 11.93 (s, 1H) 9.48 (br. s., 1H) 8.23 (s, 1H) 7.93
(s, 1H) 7.58 (d, J=7.83 Hz, 1H) 7.50 (d, J=3.79 Hz, 2H) 7.24 (s,
1H) 7.12 (dt, J=7.83, 4.17 Hz, 1H) 6.40 (br. s., 1H) 4.33 (dt,
J=13.14, 6.57 Hz, 1H) 3.71 (s, 3H) 1.78 (s, 3H) 1.26 (d, J=6.57 Hz,
6H).
Example 50
2-({5-Chloro-2-[(1-ethyl-4-methyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amino-
)-N-(methyloxy)benzamide
##STR00104##
[0218] A microwave tube was charged with
2-[(2,5-dichloro-4-pyridinyl)amino]-N-(methyloxy)benzamide (300 mg,
0.96 mmol), 1-ethyl-4-methyl-1H-pyrazol-5-amine (253 mg, 2.01
mmol), cesium carbonate (939 mg, 2.88 mmol) and DMF (7 ml). The
reaction mixture was degassed under nitrogen for 10 min and
palladium (II) acetate (10.8 mg, 0.05 mmol) and BINAP (59.8 mg,
0.096 mmol) were added. The reaction mixture was heated in an oil
bath at 90.degree. C. for 5 hrs and then in a microwave at
150.degree. C. for 50 min. It was evaporated and the residue was
dissolved in MeOH, filtered thru celite and thru an Acrodisc and
purified further on preparative Agilent HPLC (5 to 95%
water:acetonitrile with 0.1% formic acid). Fractions were combined
and evaporated. The brown oil residue was diluted in DMF and water
was added. A precipitate crashed out. It was filtered off and dried
under vacuum at 40.degree. C. for 5 hrs. LC-MS [M+H].sup.+=401.1.
1H NMR (400 MHz, DMSO-d.sub.6) ppm 11.92 (s, 1H) 9.48 (br. s., 1H)
8.27 (s, 1H) 7.94 (s, 1H) 7.59 (d, J=7.58 Hz, 1H) 7.51 (d, J=3.54
Hz, 2H) 7.22 (s, 1H) 7.09-7.16 (m, 1H) 6.40 (s, 1H) 3.85 (q, J=7.33
Hz, 2H) 3.71 (s, 3H) 1.79 (s, 3H) 1.21 (t, J=7.20 Hz, 3H).
Example 51
2-[(5-Chloro-2-{[4-methyl-1-(2-methylpropyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-N-(methyloxy)benzamide
##STR00105##
[0220] A microwave tube was charged with
2-[(2,5-dichloro-4-pyridinyl)amino]-N-(methyloxy)benzamide (300 mg,
0.96 mmol), 4-methyl-1-(2-methylpropyl)-1H-pyrazol-5-amine (309 mg,
2.01 mmol), cesium carbonate (939 mg, 2.88 mmol) and DMF (5 ml).
The reaction mixture was degassed under nitrogen for 10 min, and
palladium (II) acetate (10.8 mg, 0.05 mmol) and BINAP (59.8 mg,
0.096 mmol) were added. The reaction mixture was heated in an oil
bath at 90.degree. C. for 5 hrs and then in a microwave at
150.degree. C. for 40 min. Solvent was evaporated and the residue
was dissolved in MeOH, filtered thru celite and thru an Acrodisc an
purified further using preparative Agilent HPLC (5 to 95%
water:acetonitrile with 0.1% formic acid). Fractions were combined
and evaporated. The brown oil residue was diluted in DMF and water
was added. A precipitate crashed out. It was filtered off and dried
under vacuum at 40.degree. C. for 5 hrs. LC-MS [M+H].sup.+=429.1.
1H NMR (400 MHz, DMSO-d.sub.6) ppm 11.93 (s, 1H) 9.48 (br. s., 1H)
8.26 (s, 1H) 7.94 (s, 1H) 7.58 (d, J=7.33 Hz, 1H) 7.44-7.51 (m, 2H)
7.23 (s, 1H) 7.08-7.16 (m, 1H) 6.38 (s, 1H) 3.70 (s, 3H) 3.65 (d,
J=7.33 Hz, 2H) 2.03 (dt, J=13.71, 6.92 Hz, 1H) 1.77 (s, 3H) 0.75
(d, J=6.57 Hz, 6H).
Example 52
2-[(5-Chloro-2-{[3-(hydroxymethyl)-1-(1-methylethyl)-1H-pyrazol-5-yl]amino-
}-4-pyridinyl)amino]-N-(methyloxy)benzamide
##STR00106##
[0222] A microwave tube was charged with
2-[(2,5-dichloro-4-pyridinyl)amino]-N-(methyloxy)benzamide (188 mg,
0.602 mmol), [5-amino-1-(1-methylethyl)-1H-pyrazol-3-yl]methanol
(140 mg, 0.90 mmol), cesium carbonate (589 mg, 1.81 mmol) and DMF
(5 ml). The reaction mixture was degassed under nitrogen for 10 min
and palladium (II) acetate (6.8 mg, 0.03 mmol) and BINAP (37.5 mg,
0.06 mmol) were added. The reaction mixture was heated in an oil
bath at 90.degree. C. for 5 hrs and then in a microwave at
150.degree. C. for 40 min. It was evaporated and the residue was
dissolved in MeOH, filtered thru celite and thru an Acrodisc and
purified further using preparative Agilent HPLC (5 to 95%
water:acetonitrile with 0.1% formic acid). Fractions were combined
and evaporated. EtOAc was added to the brown oil residue. Then the
mixture was heated and hexane was added dropwise. A light yellow
precipitate crashed out upon sonication. It was filtered off and
dried under vacuum at 40.degree. C. for 12 hrs. LC-MS
[M+H].sup.+=431.2. 1H NMR (400 MHz, DMSO-d.sub.6) ppm 11.93 (br.
s., 1H) 9.54 (br. s., 1H) 8.54 (s, 1H) 7.96-8.01 (m, 1H) 7.52-7.62
(m, 3H) 7.11-7.16 (m, 1H) 6.70 (s, 1H) 6.10 (s, 1H) 4.95 (t, J=5.81
Hz, 1H) 4.43 (dt, J=13.14, 6.57 Hz, 1H) 4.33 (d, J=5.56 Hz, 2H)
3.71 (s, 3H) 1.29 (d, J=6.57 Hz, 6H).
Intermediate 9
2-(2-Chloro-5-cyclopropyl-pyridin-4-ylamino)-N-methyl-benzamide
1
##STR00107##
[0224] A solution of cyclopropylboronic acid (0.38 g, 4.40 mmole,
1.5 eq) in toluene (100 mL) was degassed with N.sub.2 at 50.degree.
C. for 15 min. To this was added Pd(PPh.sub.3).sub.4 (0.17 g, 0.15
mmole, 0.05 eq) and
2-(5-bromo-2-chloro-pyridin-4-ylamino)-N-methyl-benzamide (1 g,
2.93 mmole, 1 eq) and the resulting reaction mixture was again
degassed for 30 min. To this a degassed solution K.sub.3PO.sub.4
(2.49 g, 11.72 mmole, 4 eq) in H.sub.2O (4 mL) was added in one
portion and the resulting reaction mixture was refluxed for
overnight. Solvent was removed under reduced pressure and the
residue was diluted with water (100 mL) and then extracted with DCM
(3.times.75 mL). Combined organic layer was dried over sodium
sulfate, filtered and concentrated under reduced pressure to yield
crude product. Crude compound was purified by column chromatography
over silica gel (60-120 mesh) using 0.5% MeOH-DCM as the eluant to
give the title compound as a pale yellow solid (0.480 g, 54%).
.sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 0.60-0.70 (m, 2H),
0.95-1.05 (m, 2H), 1.60-1.71 (m, 1H), 2.76 (d, 3H, J=4.48 Hz), 7.09
(s, 1H), 7.10-7.18 (m, 1H), 7.49-7.60 (m, 2H), 7.68-7.70 (d, 1H,
J=7.64 Hz), 7.93 (s, 1H), 8.60-8.70 (m, 1H), 10.22 (s, 1H). LC-MS
[M+H].sup.+=302.0.
Example 53
2-[5-Cyclopropyl-2-(2,5-dimethyl-2H-pyrazol-3-ylamino)-pyridin-4-ylamino]--
N-methyl-benzamide
##STR00108##
[0226] To a 10 mL microwave tube were added
2-(2-chloro-5-cyclopropyl-pyridin-4-ylamino)-N-methyl-benzamide
(0.075 g, 0.25 mmol, 1 eq), 2,5-dimethyl-2H-pyrazol-3-ylamine
(0.030 g, 0.27 mmol, 1.1 eq), Cs.sub.2CO.sub.3 (0.23 g, 0.70 mmol,
2.8 eq) and 1,4-dioxane (4 mL) and resulting mixture was degassed
with N.sub.2 for 30 minutes. To this was added Pd.sub.2(dba).sub.3
(0.008 g, 0.007 mmol, 0.03 eq) and xanthphos (0.009 g, 0.014 mmol,
0.06 eq) and reaction mixture was degassed again with N.sub.2 for
another 15 minutes. The resulting reaction mixture was irradiated
in a CEM microwave at 110.degree. C. and 150 W for 40 min. The
progress of reaction was monitored by LCMS. After completion of
reaction, Cs.sub.2CO.sub.3 was removed by filtration and the
filtrate was concentrated under reduced pressure to give a crude
product. The crude compound was purified by column chromatography
over neutral alumina using 0.1% MeOH-DCM as the eluant. Solid
compound so obtained was washed with diethyl ether and pentane to
give the title compound as a yellowish solid (16 mg, 17%).
.sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 0.52-0.59 (m, 2H),
0.89-0.99 (m, 2H), 1.52-1.62 (m, 1H), 2.05 (s, 3H), 2.76-2.77 (d,
3H, J=4.44 Hz), 3.52 (s, 3H), 5.96 (s, 1H), 6.69 (s, 1H), 7.01-7.05
(t, 1H, J=7.48 Hz), 7.42-7.45 (m, 1H), 7.51-7.60 (m, 1H) 7.64-7.66
(d, 1H, J=7.32 Hz), 7.72 (s, 1H), 8.37 (s, 1H), 8.55-8.65 (brs,
1H), 9.96 (s, 1H). LC-MS [M+H].sup.+=377.2.
Example 54
2-[5-Cyclopropyl-2-(2-ethyl-5-methyl-2H-pyrazol-3-ylamino)-pyridin-4-ylami-
no]-N-methylbenzamide
##STR00109##
[0228] To a 10 mL microwave tube were added
2-(2-chloro-5-cyclopropyl-pyridin-4-ylamino)-N-methyl-benzamide
(0.075 g, 0.25 mmol, 1 eq), 2-ethyl-5-methyl-2H-pyrazol-3-ylamine
(0.035 g, 0.27 mmol, 1.1 eq), Cs.sub.2CO.sub.3 (0.23 g, 0.70 mmol,
2.8 eq) and 1,4-dioxane (3 mL) and the resulting mixture was
degassed with N.sub.2 for 30 minutes. To this mixture was added
Pd.sub.2(dba).sub.3 (0.008 g, 0.007 mmol, 0.03 eq) and xanthphos
(0.009 g, 0.015 mmol, 0.06 eq) and this mixture was degassed again
with N.sub.2 for another 10 minutes. The resulting reaction mixture
was irradiated in a CEM microwave at 110.degree. C. and 150 W for
45 min. The progress of the reaction was monitored by LCMS. After
completion of the reaction, Cs.sub.2CO.sub.3 was removed by
filtration and the filtrate was concentrated under reduced pressure
to give a crude product. It was purified using column
chromatography over neutral alumina using 0.2% MeOH-DCM as the
eluant. The solid compound so obtained was washed with diethyl
ether and hexane to give the title compound as a white solid (20
mg, 20%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 0.50-0.62
(m, 2H), 0.88-0.99 (m, 2H), 1.15-1.28 (m, 3H), 1.55-1.65 (m, 1H),
2.07 (s, 3H), 2.76-2.77 (d, 3H, J=4.36 Hz), 3.82-3.95 (m, 2H), 5.95
(s, 1H), 6.68 (s, 1H), 6.99-7.10 (m, 1H), 7.42-7.50 (m, 1H),
7.50-7.55 (m, 1H) 7.64-7.66 (d, 1H, J=7.6 Hz), 7.71 (s, 1H), 8.30
(s, 1H), 8.55-8.70 (brs, 1H), 9.95 (s, 1H). LC-MS
[M+H].sup.+=391.4.
Intermediate 10
2-(5-Bromo-2-chloro-pyridin-4-ylamino)-N-methoxy-benzamide
##STR00110##
[0230] A mixture of 5-bromo-2-chloro-4-iodo-pyridine (5 g, 15.72
mmole, 1 eq), 2-amino-N-methoxy-benzamide (2.61 g, 15.72 mmole, 1
eq) and K.sub.3PO.sub.4 (8.34 g, 39.3 mmole, 2.5 eq), and
1,4-dioxane (30 mL) was degassed with N.sub.2 for 1 h. To this was
added DPEPhos (0.67 g, 1.25 mmole, 0.08 eq) and Pd(OAc).sub.2 (0.07
g, 0.31 mmole, 0.02 eq) and this mixture was degassed again with
N.sub.2 for 30 min. The resulting mixture was refluxed for
overnight. After completion of reaction, solvent was removed under
reduced pressure and residue was diluted with water (100 mL) and
extracted with 5% MeOH-DCM (3.times.100 mL). Combined organic
layers were dried over sodium sulfate, filtered and concentrated
under reduced pressure to give a crude compound. It was purified by
column chromatography over silica gel (60-120 mesh) using 20% Ethyl
acetate-hexane as the eluant to yield the title compound as off
white solid (3.5 g, 62%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6):
.delta. 3.56 (s, 3H), 6.88-6.98 (m, 1H), 7.08 (s, 1H), 7.17-7.25
(m, 1H), 7.26-7.35 (m, 1H), 7.82-7.90 (m, 1H), 8.23 (s, 1H), 12.86
(brs, 1H). LC-MS [M+H].sup.+=356.30.
Intermediate 11
2-(2-Chloro-5-cyclopropyl-pyridin-4-ylamino)-N-methoxy-benzamide
##STR00111##
[0232] In a sealable tube toluene (50 mL) was degassed with N.sub.2
at 50.degree. C. for 15 min and to this
2-(5-bromo-2-chloro-pyridin-4-ylamino)-N-methoxy-benzamide (1.5 g,
4.21 mmole, 1 eq), cyclopropylboronic acid (1.4 g, 16.85 mmole, 4
eq) and Pd(PPh.sub.3).sub.4 (0.24 g, 0.21 mmole, 0.05 eq) were
added and resulting mixture was degassed for 30 min. To this NaBr
(0.44 g, 4.33 mmole, 1.03 eq) and a solution of KF (0.8 g, 13.90
mmole, 3.3 eq) in H.sub.2O (3 mL) were added; again degassed with
N.sub.2 for 15 min. The tube was sealed and the resulting mixture
was heated at 100.degree. C. for 24 h. After completion of
reaction, reaction mixture was allowed to cool at room temperature,
poured into water (100 mL) and extracted with toluene (2.times.50
mL). Combined organic layer was dried over sodium sulfate, filtered
and concentrated under reduced pressure. The crude compound was
purified by washing with 0.5% DCM-Et.sub.2O to give titled compound
as pale yellow solid (0.7 g, 53%). .sup.1H-NMR (400 MHz,
DMSO-d.sub.6): .delta. 0.57-072 (m, 2H), 0.97-1.10 (m, 2H),
1.62-1.75 (m, 1H), 3.68 (s, 3H), 7.02 (s, 1H), 7.11-7.20 (m, 1H),
7.51-7.67 (m, 3H), 7.94 (s, 1H), 9.62 (s, 1H), 11.92 (brs, 1H).
LC-MS [M+H].sup.+=318.2.
Example 55
2-[5-Cyclopropyl-2-(2,5-dimethyl-2H-pyrazol-3-ylamino)-pyridin-4-ylamino]--
N-methoxy-benzamide
##STR00112##
[0234] To a 10 mL microwave tube were added
2-(2-chloro-5-cyclopropyl-pyridin-4-ylamino)-N-methoxy-benzamide
(0.075 g, 0.24 mmol, 1 eq), 2,5-dimethyl-2H-pyrazol-3-ylamine (0.05
g, 0.47 mmol, 2 eq), Cs.sub.2CO.sub.3 (0.23 g, 0.71 mmol, 3 eq),
and 1,4-dioxane (3 mL). The resulting mixture was degassed with
N.sub.2 for 15 min. To this was added Pd.sub.2(dba).sub.3 (0.015 g,
0.014 mmol, 0.06 eq) and xanthphos (0.03 g, 0.06 mmol, 0.25 eq) and
the mixture was again degassed with N.sub.2 for 30 min. The
resulting mixture was irradiated in a CEM microwave at 120.degree.
C., 150 W for 35 min. After completion of reaction, solvent was
removed under reduced pressure and crude compound was purified
using column chromatography over silica gel (100-200 mesh) using 1%
MeOH-DCM as the eluant followed by prep HPLC. Solid compound so
obtained was washed with diethyl ether and pentane to give the
title compound as an off white solid (11 mg, 12%). .sup.1H-NMR (400
MHz, DMSO-d.sub.6): .delta. 0.52-0.60 (m, 2H), 0.82-1.00 (m, 2H),
1.50-1.62 (m, 1H), 2.05 (s, 3H), 3.52 (s, 3H), 3.70 (s, 3H), 5.96
(s, 1H), 6.67 (s, 1H), 6.98-7.10 (m, 1H), 7.40-7.60 (m, 3H), 7.73
(s, 1H), 8.39 (s, 1H), 9.46 (s, 1H), 11.88 (brs, 1H). LC-MS
[M+H].sup.+=393.4.
Example 56
2-[5-Cyclopropyl-2-(2-ethyl-5-methyl-2H-pyrazol-3-ylamino)-pyridin-4-ylami-
no]-N-methoxy-benzamide
##STR00113##
[0236] To a 10 mL microwave tube were added
2-(2-chloro-5-cyclopropyl-pyridin-4-ylamino)-N-methoxy-benzamide
(0.075 g, 0.24 mmol, 1 eq), 2-ethyl-5-methyl-2H-pyrazol-3-ylamine
(0.05 g, 0.36 mmol, 1.5 eq), Cs.sub.2CO.sub.3 (0.23 g, 0.71 mmol, 3
eq), and 1,4-dioxane (3 mL) and the resulting mixture was degassed
with N.sub.2 for 15 min. To this was added Pd.sub.2(dba).sub.3
(0.014 g, 0.014 mmol, 0.06 eq) and xanthphos (0.03 g, 0.06 mmol,
0.25 eq) and this mixture was again degassed with N.sub.2 for
another 10 min. The resulting mixture was irradiated in a CEM
microwave at 120.degree. C., 150 W for 35 min. After the reaction
was finished solvent was removed under reduced pressure and crude
product was purified by column chromatography over silica gel
(100-200 mesh) using 1% MeOH-DCM as the eluant followed by washing
with diethyl ether and pentane and DCM to give the title compound
as an off white solid (5 mg, 5%). .sup.1H-NMR (400 MHz,
DMSO-d.sub.6): .delta. 0.50-0.60 (m, 2H), 0.88-1.00 (m, 2H),
1.15-1.27 (m, 3H), 1.55-1.65 (m, 1H), 2.07 (s, 3H), 3.70 (s, 3H),
3.80-3.95 (m, 2H), 5.95 (s, 1H), 6.64 (s, 1H), 6.90-7.10 (m, 1H),
7.40-7.60 (m, 3H), 7.72 (s, 1H), 8.34 (s, 1H), 9.47 (s, 1H), 11.88
(brs, 1H). LC-MS [M+H].sup.+=407.3.
Intermediate 12
2-(2-Chloro-5-isopropenyl-pyridin-4-ylamino)-N-methyl-benzamide
##STR00114##
[0238] In a tube a solution of isopropenyboronic acid pinacol ester
(3.31 mL, 17.6 mmole, 3 eq) in toluene (200 mL) was degassed with
N.sub.2 at 50.degree. C. for 15 min. To this was added
Pd(PPh.sub.3).sub.4 (0.68 g, 0.59 mmole, 0.1 eq) and
2-(5-bromo-2-chloro-pyridin-4-ylamino)-N-methyl-benzamide (2 g,
5.87 mmole, 1 eq). This mixture was again degassed with N.sub.2 for
30 min. A degassed solution of K.sub.3PO.sub.4 (4.98 g, 23.48
mmole, 4 eq) in H.sub.2O (8 mL) was added to the above mixture in
one portion and the resulting mixture was heated at 110.degree. C.
for overnight. After completion of the reaction, it was allowed to
cool to room temperature and solvent was removed under reduced
pressure to yield crude product. It was purified by column
chromatography over silica gel (60-120 mesh) using 0.5% MeOH-DCM as
the eluant to give the title compound as an off white solid (0.8 g,
45%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 2.06 (s, 3H),
2.73-2.74 (d, 3H, J=4.52 Hz), 5.14 (s, 1H), 5.46 (s, 1H), 7.04-7.15
(m, 2H), 7.45-7.60 (m, 2H), 7.64-7.71 (m, 1H), 7.96 (s, 1H),
8.60-8.72 (brs, 1H), 9.87 (s, 1H). LC-MS [M+H].sup.+=302.2.
Example 57
2-[2-(2,5-Dimethyl-2H-pyrazol-3-ylamino)-5-isopropenyl-pyridin-4-ylamino]--
N-methyl-benzamide
##STR00115##
[0240] To a 10 mL microwave tube was added
2-(2-chloro-5-isopropenyl-pyridin-4-ylamino)-N-methyl-benzamide
(0.075 g, 0.25 mmol, 1 eq), 2,5-dimethyl-2H-pyrazol-3-ylamine
(0.055 g, 0.50 mmol, 2 eq), Cs.sub.2CO.sub.3 (0.24 g, 0.74 mmol, 3
eq) and 1,4-dioxane (3 mL). The resulting mixture was degassed with
N.sub.2 for 30 minutes. Then Pd(OAc).sub.2 (0.015 g, 0.07 mmol,
0.27 eq) and BINAP (0.046 g, 0.074 mmol, 0.3 eq) were added and the
mixture degassed again with N.sub.2 for another 10 minutes. The
resulting reaction mixture was irradiated in a CEM microwave at
110.degree. C. and 150 W for 45 min. The progress of the reaction
was monitored by LCMS. After it was complete, Cs.sub.2CO.sub.3 was
removed by filtration and the filtrate was concentrated under
reduced pressure. The resulting crude product was purified by
column chromatography over neutral alumina using 0.2% MeOH-DCM as
the eluant. This gave the title compound as an off white solid (180
mg, 48%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 2.02-2.10
(m, 6H), 2.73-2.74 (d, 3H, J=4.48 Hz), 3.53 (s, 3H), 5.05 (s, 1H),
5.31 (s, 1H), 5.97 (s, 1H), 6.67 (s, 1H), 6.90-7.05 (m, 1H),
7.4-7.55 (m, 2H), 7.61-7.63 (d, 1H, J=7.04 Hz), 7.76 (s, 1H), 8.48
(s, 1H), 8.54-8.67 (brs, 1H), 9.61 (s, 1H). LC-MS
[M+H].sup.+=377.4.
Example 58
2-[2-(2,5-Dimethyl-2H-pyrazol-3-ylamino)-5-isopropyl-pyridin-4-ylamino]-N--
methyl-benzamide
##STR00116##
[0242] A solution of
2-[2-(2,5-dimethyl-2H-pyrazol-3-ylamino)-5-isopropenyl-pyridin-4-ylamino]-
-N-methyl-benzamide (0.13 g, 0.345 mmole, 1 eq) in ethanol (10 mL)
was degassed with N.sub.2 and to this PtO.sub.2 (0.012 g, 0.052
mmole, 0.15 eq) was added. The resulting mixture was stirred at
room temperature under H.sub.2 atmosphere with balloon pressure for
10 h. After completion of the reaction, it was filtered through a
celite bed which was then washed with ethanol (10 mL). The filtrate
was evaporated under reduced pressure to give a solid residue which
was purified by washing with diethyl ether to give the title
compound as a gray solid (80 mg, 61%). .sup.1H-NMR (400 MHz,
DMSO-d.sub.6): .delta. 1.24-1.26 (d, 6H, J=6.76 Hz), 2.05 (s, 3H),
2.75-2.76 (d, 3H, J=4.48 Hz), 2.89-3.10 (m, 1H), 3.53 (s, 3H), 5.96
(s, 1H), 6.67 (s, 1H), 6.85-7.10 (m, 1H), 7.40-7.50 (m, 2H),
7.65-7.67 (d, 1H, J=7.76 Hz), 7.86 (s, 1H), 8.36 (s, 1H), 8.60-8.70
(brs, 1H), 9.89 (s, 1H). LC-MS [M+H].sup.+=379.2.
Example 59
2-({5-Chloro-2-[(1,5-dimethyl-1H-pyrazol-4-yl)amino]-4-pyridinyl}amino)-N--
methylbenzamide
##STR00117##
[0244] To a 10 mL sealable tube was added
2-[(2,5-dichloro-4-pyridinyl)amino]-N-methylbenzamide (95 mg, 0.321
mmol), 1,5-dimethyl-1H-pyrazol-4-amine (35.7 mg, 0.321 mmol), BINAP
(20 mg, 0.032 mmol), cesium carbonate (314 mg, 0.964 mmol), and
palladium(II) acetate (7.21 mg, 0.032 mmol) in 1,4-dioxane (5 mL).
The reaction vessel was sealed and heated at 150.degree. C. for 1
hr. The reaction mixture was purified using a prep HPLC (0.1%
formic acid, 5 to 95% water:acetonitrile). Fractions were combined
and evaporated.
2-({5-Chloro-2-[(1,5-dimethyl-1H-pyrazol-4-yl)amino]-4-pyridinyl}amino)-N-
-methylbenzamide (16 mg, 0.037 mmol, 11.38% yield) was isolated as
yellow oil LCMS (M+H).sup.+=371.1; 1H NMR (400 MHz, MeOD) ppm 2.17
(s, 3H) 2.91 (s, 3H) 3.78 (s, 3H) 6.47 (s, 1H) 7.13-7.24 (m, 1H)
7.40 (s, 1H) 7.46-7.57 (m, 2H) 7.67 (dd, J=7.83, 1.26 Hz, 1H) 7.82
(s, 1H).
Example 60
2-{[5-Chloro-2-({1-ethyl-3-[(methyloxy)methyl]-1H-pyrazol-5-yl}amino)-4-py-
ridinyl]amino}-N-methylbenzamide
60a) 1-Ethyl-3-[(methyloxy)methyl]-1H-pyrazol-5-amine
##STR00118##
[0246] To a 25 mL round bottom was added
4-(methyloxy)-3-oxobutanenitrile (500 mg, 4.42 mmol), ethyl
hydrazine (655 mg, 4.42 mmol), and 2 M HCl (2.210 mL, 4.42 mmol) in
ethanol (10 mL). The reaction mixture was stirred at the room
temperature overnight. The reaction mixture was then evaporated and
pardoned between 20 mL of ethyl acetate and 20 mL of 1 M
Na.sub.2CO.sub.3. The organic layer was washed with brine, filtered
and evaporated to obtain
1-ethyl-3-[(methyloxy)methyl]-1H-pyrazol-5-amine (450 mg, 2.465
mmol, 55.8% yield) as a yellow oil. The product was used in the
next step without further purification. LCMS (M+H).sup.+=156.1; 1H
NMR (400 MHz, DMSO-d.sub.6) ppm 1.20 (t, J=7.20 Hz, 3H) 3.19 (s,
3H) 3.82 (q, J=7.33 Hz, 2H) 4.11 (s, 2H) 5.11 (s, 2H) 5.22 (s,
1H).
60b)
2-{[5-chloro-2-({1-ethyl-3-[(methyloxy)methyl]-1H-pyrazol-5-yl}amino)-
-4-pyridinyl]amino}-N-methylbenzamide
##STR00119##
[0248] The title compound was prepared substantially as described
in Example 59 except using
1-ethyl-3-[(methyloxy)methyl]-1H-pyrazol-5-amine instead of
5-amino-1-methyl-1H-pyrazole. LCMS (M+H).sup.+=415.1 (M+H); 1H NMR
(400 MHz, DMSO-d.sub.6) ppm 1.24 (t, J=7.20 Hz, 3H) 2.78 (d, J=4.55
Hz, 3H) 3.23 (s, 3H) 3.95 (q, J=7.07 Hz, 2H) 4.25 (s, 2H) 6.20 (s,
1H) 6.77 (s, 1H) 7.05-7.16 (m, 1H) 7.44-7.60 (m, 2H) 7.70 (dd,
J=7.83, 1.26 Hz, 1H) 8.01 (s, 1H) 8.63 (s, 1H) 8.69 (q, J=4.29 Hz,
1H) 10.12 (s, 1H).
Example 61
2-[(5-Chloro-2-{[3-[(ethyloxy)methyl]-1-(1-methylethyl)-1H-pyrazol-5-yl]am-
ino}-4-pyridinyl)amino]-N-methylbenzamide
61a) 3-[(Ethyloxy)methyl]-1-(1-methylethyl)-1H-pyrazol-5-amine
##STR00120##
[0250] To a 25 mL round bottom was added
4-(ethyloxy)-3-oxobutanenitrile (550 mg, 4.33 mmol), isopropyl
hydrazine hydrochloride (478 mg, 4.33 mmol), and HCl (2.163 mL,
4.33 mmol) in Ethanol (10 mL). The reaction mixture was stirred at
the room temperature overnight. The reaction mixture was then
evaporated, and partitioned between 20 mL of ethyl acetate and 20
mL of 1 M Na.sub.2CO.sub.3. The organic layer was washed with
brine, filtered and evaporated.
3-[(Ethyloxy)methyl]-1-(1-methylethyl)-1H-pyrazol-5-amine (520 mg,
2.84 mmol, 65.6% yield) was isolated as yellow oil. The product was
used in the next step without further purification. 1H NMR (400
MHz, DMSO-d.sub.6) ppm 1.09 (t, J=7.07 Hz, 3H) 1.26 (d, J=6.57 Hz,
6H) 3.40 (q, J=6.91 Hz, 2H) 4.16 (s, 2H) 4.32 (quin, J=6.57 Hz, 1H)
5.06 (s, 2H) 5.22 (s, 1H)
61b)
2-[(5-Chloro-2-{[3-[(ethyloxy)methyl]-1-(1-methylethyl)-1H-pyrazol-5--
yl]amino}-4-pyridinyl)amino]-N-methylbenzamide
##STR00121##
[0252] The title compound was prepared substantially as described
in Example 59 except using
3-[(ethyloxy)methyl]-1-(1-methylethyl)-1H-pyrazol-5-amine instead
of 5-amino-1-methyl-1H-pyrazole. LCMS (ES) m/z=443.1 (M+H); 1H NMR
(400 MHz, MeOD) ppm 1.21 (t, J=7.07 Hz, 3H) 1.40 (d, J=6.82 Hz, 6H)
2.91 (s, 3H) 3.56 (q, J=6.91 Hz, 2H) 4.45 (s, 2H) 4.53 (quin,
J=6.69 Hz, 1H) 6.17 (s, 1H) 6.60 (s, 1H) 7.05-7.18 (m, 1H)
7.40-7.56 (m, 2H) 7.62-7.70 (m, 1H) 7.92 (s, 1H)
Example 62
2-{[5-Chloro-2-({1-ethyl-3-[(ethyloxy)methyl]-1H-pyrazol-5-yl}amino)-4-pyr-
idinyl]amino}-N-methylbenzamide
62a) 1-Ethyl-3-[(ethyloxy)methyl]-1H-pyrazol-5-amine
##STR00122##
[0254] To a 25 mL round bottom was added
4-(ethyloxy)-3-oxobutanenitrile (550 mg, 4.33 mmol), ethyl
hydrazine oxalate (641 mg, 4.33 mmol), and HCl (2.163 mL, 4.33
mmol) in ethanol (10 mL). The reaction mixture was stirred at the
room temperature overnight. The reaction mixture was then
evaporated, and partitioned between 20 mL of ethyl acetate and 20
mL of 1 M Na.sub.2CO.sub.3. The organic layer was washed with
brine, filtered and evaporated.
1-Ethyl-3-[(ethyloxy)methyl]-1H-pyrazol-5-amine (420 mg, 2.234
mmol, 51.6% yield) was isolated as yellow oil. The product was used
in the next step without further purification. LCMS
(M+H).sup.+=170.1 (M+H); 1H NMR (400 MHz, DMSO-d.sub.6) ppm 1.08
(t, J=6.95 Hz, 3H) 1.19 (t, J=7.07 Hz, 3H) 3.37-3.43 (m, 2H) 3.81
(q, J=7.24 Hz, 2H) 4.15 (s, 2H) 5.09 (s, 2H) 5.22 (s, 1H).
62b)
2-{[5-Chloro-2-({1-ethyl-3-[(ethyloxy)methyl]-1H-pyrazol-5-yl}amino)--
4-pyridinyl]amino}-N-methylbenzamide
##STR00123##
[0256] The title compound was prepared substantially as described
in Example 59 except using
1-ethyl-3-[(ethyloxy)methyl]-1H-pyrazol-5-amine instead of
5-amino-1-methyl-1H-pyrazole. LCMS (M+H).sup.+=429.1 (M+H); 1H NMR
(400 MHz, MeOD) ppm 1.21 (t, J=7.07 Hz, 3H) 1.34 (t, J=7.33 Hz, 3H)
2.91 (s, 3H) 3.55 (q, J=6.91 Hz, 2H) 4.04 (q, J=7.33 Hz, 2H) 4.43
(s, 2H) 6.22 (s, 1H) 6.66 (s, 1H) 7.10-7.20 (m, 1H) 7.45-7.53 (m,
1H) 7.53-7.58 (m, 1H) 7.66 (dd, J=7.83, 1.52 Hz, 1H) 7.93 (s,
1H).
Example 63
2-({5-Chloro-2-[(1-ethyl-3-methyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amino-
)-N-hydroxy-N-methylbenzamide
63a)
2-({5-Chloro-2-[(1-ethyl-3-methyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}-
amino)benzonitrile
##STR00124##
[0258] To a 50 mL tube was added
2-[(2,5-dichloro-4-pyridinyl)amino]benzonitrile (738 mg, 2.80
mmol), 1-ethyl-3-methyl-1H-pyrazol-5-amine (350 mg, 2.80 mmol),
BINAP (696 mg, 1.118 mmol), cesium carbonate (2733 mg, 8.39 mmol)
and palladium(II) acetate (62.8 mg, 0.280 mmol) in 1,4-dioxane (15
mL). The reaction mixture was heated to 120.degree. C. for 18 hr.
Solid was filtered off and discarded and the solution was
concentrated. Product was then dissolved in 1 M HCl (1 mL) and
extracted with EtOAc. The organic layer was discarded. The water
layer was neutralized with 1 M NaOH (to pH 8) and extracted with
EtoAc. The organic layer was separated, washed with brine, dried
over MgSO.sub.4, filtered and evaporated. The solid was the
dissolved in 20 mL of EtOAc and 20 mL of water and 1 mL of acetic
acid were added. The organic layer was separated, washed with
brine, dried over MgSO.sub.4, filtered and evaporated.
2-({5-Chloro-2-[(1-ethyl-3-methyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amin-
o)benzonitrile (450 mg, 1.275 mmol, 45.6% yield) was isolated as
orange foam. This product was used in the next step without further
purification. LCMS (M+H).sup.+=353.1; 1H NMR (400 MHz,
DMSO-d.sub.6) ppm 1.19 (t, J=7.20 Hz, 3H) 1.91 (s, 3H) 3.84 (q,
J=7.24 Hz, 2H) 5.92 (s, 1H) 6.03 (s, 1H) 7.41 (td, J=7.64, 0.88 Hz,
1H) 7.48 (d, J=8.08 Hz, 1H) 7.76 (td, J=7.83, 1.52 Hz, 1H) 7.91
(dd, J=7.83, 1.52 Hz, 1H) 7.96 (s, 1H) 8.47 (d, J=7.07 Hz, 1H)
11.98 (br. s., 1H)
63b)
2-({5-Chloro-2-[(1-ethyl-3-methyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}-
amino)benzoic acid
##STR00125##
[0260] To a 50 mL round bottom flask was added
2-({5-chloro-2-[(1-ethyl-3-methyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amin-
o)benzonitrile (300 mg, 0.850 mmol), and NaOH 1 M solution (10 ml,
10.00 mmol) in 1,4-dioxane (10 mL). The mixture was heated under
reflux for 18 hours. Ethyl acetate was added (20 mL) and the layers
separated--all product stayed in the water phase. The water phase
was neutralized with 6 N HCl, and 40 mL of ethyl acetate were
added. The organic layer was separated, washed with brine, dried
over MgSO.sub.4, filtered and evaporated.
2-({5-Chloro-2-[(1-ethyl-3-methyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amin-
o)benzoic acid (220 mg, 0.562 mmol, 66.1% yield) was isolated as
off-white solid and used in the next step without further
purification. LCMS (M+H).sup.+=372.1; 1H NMR (400 MHz, MeOD) ppm
1.33 (t, J=7.20 Hz, 4H) 2.22 (s, 3H) 4.00 (q, J=7.33 Hz, 2H) 6.01
(s, 1H) 6.82 (s, 1H) 7.06 (td, J=7.52, 1.14 Hz, 1H) 7.39-7.49 (m,
1H) 7.49-7.55 (m, 1H) 7.87-7.94 (m, 1H) 8.05 (dd, J=7.83, 1.52 Hz,
1H).
63c)
2-({5-Chloro-2-[1-ethyl-3-methyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}a-
mino)-N-hydroxy-N-methylbenzamide
##STR00126##
[0262] A solution of
2-({5-chloro-2-[(1-ethyl-3-methyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amin-
o)benzoic acid (55 mg, 0.148 mmol), HOBT (22.65 mg, 0.148 mmol) and
EDC (28.4 mg, 0.148 mmol) in N,N-dimethylformamide (DMF) (5 mL) was
stirred under nitrogen at room temp for 30 minutes. To this
solution was added N-methylhydroxylamine (12.35 mg, 0.148 mmol) and
the solution was stirred for another 15 minutes. The reaction
mixture was cooled to 5.degree. C. and DIEA (0.052 mL, 0.296 mmol)
was added dropwise. After addition, the reaction mixture was
stirred at room temperature overnight. The reaction mixture was
purified using preparative HPLC (0.1% formic acid, 5 to 95%
water:acetonitrile). Fractions were combined and evaporated.
2-({5-Chloro-2-[(1-ethyl-3-methyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amin-
o)-N-hydroxy-N-methylbenzamide (16 mg, 0.034 mmol, 23.05% yield)
isolated as a white solid. LCMS (M+H).sup.+=401.0 (M+H); 1H NMR
(400 MHz, MeOD) ppm 1.30 (t, J=7.20 Hz, 3H) 2.19 (s, 3H) 3.37 (d,
J=1.52 Hz, 3H) 3.96 (q, J=7.07 Hz, 2H) 5.95 (s, 1H) 6.32 (s, 1H)
7.21-7.33 (m, 1H) 7.42-7.53 (m, 2H) 7.56 (d, J=7.33 Hz, 1H) 7.86
(s, 1H).
Example 64
2-({5-Chloro-2-[(1-ethyl-3-methyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amino-
)-N-(ethyloxy)benzamide
##STR00127##
[0264] The title compound was prepared substantially as described
in Example 63 except using O-ethylhydroxylamine hydrochloride
instead of N-methylhydroxylamine. LCMS (M+H).sup.+=415.1 (M+H); 1H
NMR (400 MHz, MeOD) ppm 1.23-1.42 (m, 6H) 2.21 (s, 3H) 3.87-4.10
(m, 4H) 5.99 (s, 1H) 6.59 (s, 1H) 7.11-7.28 (m, 1H) 7.46-7.71 (m,
3H) 7.92 (s, 1H).
Intermediate 13
6-Chloro-4-iodo-nicotinonitrile
##STR00128##
[0266] To a mixture of THF (100 mL) and hexane (40 mL) under
nitrogen atmosphere was added DIPA (22.21 mL, 158.78 mmole, 1.1
eq). The mixture was cooled to -80.degree. C. and to this was added
n-BuLi (63.57 mL, 158.78 mmole, 1.1 eq) dropwise. After completion
of addition the resulting mixture was allowed to warm and stirred
at -10.degree. C. for 15 min. The reaction mixture was again cooled
to -80.degree. C. and a solution of 6-chloro-nicotinonitrile (20 g,
144.35 mmol, 1 eq) in THF (100 mL) was added dropwise. The
resulting mixture was stirred at -80.degree. C. for 1 h. After 1 h
a solution of iodine (43.96 g, 173.22 mmole, 1.2 eq) in THF (100
mL) was added in one portion. After completion of the reaction, the
reaction was quenched with water (100 mL) and extracted with
diethyl ether (6.times.100 mL). Combined organic layers were washed
with saturated solution of sodium thiosulfate (2.times.100 mL),
dried over sodium sulfate, filtered and concentrated under vacuum
to give crude product. Crude compound was purified by column
chromatography over silica gel (100-200 mesh) using 2% ethyl
acetate-hexane as the eluant to yield the title compound as a
faintly yellow solid (15 g, 39%). .sup.1H-NMR (400 MHz,
DMSO-d.sub.6): .delta. 8.38 (s, 1H), 8.79 (s, 1H). LC-MS calculated
for C.sub.6H.sub.2ClIN.sub.2 (M+H) 264.90. found 264.9.
Intermediate 14
2-(2-Chloro-5-cyanopyridin-4-ylamino)-N-methylbenzamide
##STR00129##
[0268] A mixture of 6-chloro-4-iodo-nicotinonitrile (14 g, 53.03
mmole, 1 eq), 2-amino-N-methoxy-benzamide (7.96 g, 53.03 mmole, 1
eq) and K.sub.3PO.sub.4 (28.14 g, 132.57 mmole, 2.5 eq) in
1,4-dioxane (250 mL) was degassed with N.sub.2 for 1 h. To this
mixture were added Pd(OAC).sub.2 (0.238 g, 1.06 mmole, 0.02 eq) and
DPEPhos (2.28 g, 4.24 mmole, 0.08 eq). The resulting reaction
mixture was degassed with N.sub.2 for another 15 min after which
the resulting reaction mixture was stirred at 110.degree. C.
overnight. After completion of reaction solid material was
collected by filtration, dissolved in water (500 mL), and extracted
with ethyl acetate (5.times.200 mL). Combined organic layers were
dried over sodium sulfate, filtered and concentrated under reduced
pressure to give crude material. It was purified by column
chromatography over silica gel (60-120 mesh) using 0.2% methanolic
ammonia (10% ammonia in MeOH) in dichloromethane as the eluant to
give the title compound as a pale yellow solid (9 g, 59%).
.sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 2.75 (d, 3H, J=4.56
Hz), 7.09 (s, 1H), 7.25-7.35 (m, 1H), 7.53-7.60 (m, 2H), 7.72 (d,
1H, J=7.48 Hz), 8.56 (s, 1H), 8.69-8.79 (brs, 1H), 10.65 (s, 1H).
LC-MS calculated for C.sub.14H.sub.11ClN.sub.4O (M+H) 287.06. found
286.9.
Intermediate 15
5-[4-(2-Hydroxyethyl)-1-piperazinyl]-N-methyl-2-nitrobenzamide
3
##STR00130##
[0270] A solution of 5-fluoro-N-methyl-2-nitrobenzamide,
1-(2-hydroxyethyl)piperazine, and Hunig's base in 20 mL of DMF was
stirred at room temperature over the weekend. The resulting mixture
was rotavaped to dryness, and the residue was purified by flash
column SF40-150 at 2%-10% MeOH/CH.sub.2Cl.sub.2. Product came out
at 7% MeOH/CH.sub.2Cl.sub.2. MS(ES) m/e 309 [M+H].
Intermediate 16
2-amino-5-[4-(2-Hydroxyethyl)-1-piperazinyl]-N-methylbenzamide
4
##STR00131##
[0272] A solution of
5-[4-(2-hydroxyethyl)-1-piperazinyl]-N-methyl-2-nitrobenzamide 3 in
250 mL of MeOH in a 250 mL round bottom flask was stirred at room
temperature under hydrogen balloon over night. The resulted mixture
was filtered to get rid of the Pd catalyst. TLC in 10%
MeOH/CH.sub.2Cl.sub.2 showed no more starting material and a major
product. The solvent was rotavaped to dryness, and the residue was
used without further purification. MS(ES) m/e 279 [M+H].
Example 65
2-({5-Cyano-2-[(1,3-dimethyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amino)-N-m-
ethylbenzamide
##STR00132##
[0274]
2-({5-Cyano-2-[(1,3-dimethyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}ami-
no)-N-methylbenzamide was synthesized substantially according to
the procedure of example 55 using Intermediate 15 and
5-amino-1,3-dimethylpryazole. .sup.1H-NMR (400 MHz, DMSO-d.sub.6):
.delta. 2.08 (d, 3H), 2.77 (d, 3H), 3.53 (s, 3H), 6.01 (s, 1H),
6.59 (s, 1H), 6.01 (s, 1H), 7.18 (t, 1H), 7.54 (t, 2H), 7.72 (d,
1H), 8.33 (s, 1H), 8.71 (d, 1H), 9.15 (s, 1H), 10.33 (s, 1H). LC-MS
(M+H).sup.+=362.0.
Example 66
2-[(2,5-Dichloro-4-pyridinyl)amino]-5-[4-(2-hydroxyethyl)-1-piperazinyl]-N-
-methylbenzamide 6
##STR00133##
[0276] A sealed tube was charged with 2,5-dichloro-4-iodopyridine,
2-amino-5-[4-(2-hydroxyethyl)-1-piperazinyl]-N-methylbenzamide 4,
and cesium carbonate in 1,4-dioxane. The reaction mixture was
degassed by nitrogen for 10 min. At same time BINAP and
palladium(II) acetate were added into it and the reaction mixture
was heated in 120.degree. C. in an oil bath over night.
[0277] TLC in 10% EtOAc/hexane showed no
2,5-dichloro-4-indopyridine. TLC in 10% MeOH/CH.sub.2Cl.sub.2
showed no
2-amino-5-[4-(2-hydroxyethyl)-1-piperazinyl]-N-methylbenzamide.
LCMS showed the reaction had a peak that could be the desired
product. While the reaction mixture temperature was maintained at
around 80.degree. C., it was filtered, and the solid was washed
with THF and CH.sub.3CN. The solid was filtered, dried by vacuum,
and purified by flash column 1-8% MeOH/CH.sub.2Cl.sub.2, to give
the captioned product as a brown oil. MS(ES) m/e 426 [M+2H].
Example 67
2-({5-Chloro-2-[(1-ethyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amino)-5-[4-(2-
-hydroxyethyl)-1-piperazinyl]-N-methylbenzamide 9
##STR00134##
[0279] A sealed tube was charged with
2-[(2,5-dichloro-4-pyridinyl)amino]-5-[4-(2-hydroxyethyl)-1-piperazinyl]--
N-methylbenzamide 6,1-ethyl-1H-pyrazol-5-amine, and cesium
carbonate in 1,4-dioxane. The reaction mixture was degassed by
nitrogen for 10 min. At same time, BINAP and palladium(II) acetate
were added and the reaction mixture was heated to 160.degree. C. in
microwave for 40 minutes. LCMS showed a peak believed to be the
desired product. The solvent was rotavaped to dryness, and the
residue was purified by HPLC to give the captioned product. MS(ES)
m/e 500 [M+H].
Example 68
2-({5-Chloro-2-[(1,3-dimethyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amino)-5--
[4-(2-hydroxyethyl)-1-piperazinyl]-N-methylbenzamide 10
##STR00135##
[0281] A sealed tube was charged with
2-[(2,5-dichloro-4-pyridinyl)amino]-5-[4-(2-hydroxyethyl)-1-piperazinyl]--
N-methylbenzamide 6,1-ethyl-1H-pyrazol-5-amine, and cesium
carbonate in 1,4-dioxane. The reaction mixture was degassed by
nitrogen for 10 min. Then BINAP and palladium(II) acetate were
added into it, and the reaction mixture was heated to 160.degree.
C. in microwave for 40 minutes. LCMS showed the a peak believed to
correspond to the desired product. The solvent was rotavaped to
dryness, and the residue was purified by HPLC to give product.
MS(ES) m/e 500 [M+H].
Example 69
4-Chloro-2-[(5-chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino-
}-4-pyridinyl)amino]-N-(methyloxy)benzamide
##STR00136##
[0282] Step 1:
[0283] To a degassed solution of 2,5-dichloro-4-iodopyridine (4.5
g, 16.43 mmol), 2-amino-4-chlorobenzonitrile (2.507 g, 16.43 mmol)
and potassium triphosphate (10.46 g, 49.3 mmol) in 1,4-dioxane (60
ml) stirred under nitrogen at the room temperature was added
DPEPhos (0.708 g, 1.314 mmol) and palladium acetate (0.148 g, 0.657
mmol). The reaction mixture was stirred at the reflux for 18 hr.
The reaction mixture was filtered. The solution was evaporated.
Ether (50 ml) was added and the formed solid was filtered.
4-chloro-2-[(2,5-dichloro-4-pyridinyl)amino]benzonitrile (2.8 g,
9.38 mmol, 57.1% yield) was isolated as an orange solid. .sup.1H
NMR (400 MHz, DMSO-d.sub.6) ppm 6.70 (s, 1H) 7.53 (dd, J=8.34, 2.02
Hz, 1H) 7.65 (d, J=2.02 Hz, 1H) 7.95 (d, J=8.34 Hz, 1H) 8.28 (s,
1H) 9.12 (br. s., 1H); HPLC Rt=3.50 min, MS (ESI): 298.0, 300.0
[M+H].sup.+.
Step 2.
[0284] A solution of
4-chloro-2-[(2,5-dichloro-4-pyridinyl)amino]benzonitrile (2.8 g,
9.38 mmol), 3-methyl-1-(1-methylethyl)-1H-pyrazol-5-amine (1.305 g,
9.38 mmol) and cesium carbonate (9.17 g, 28.1 mmol) in 1,4-dioxane
(40 mL) was degassed. DPEPhos (0.404 g, 0.750 mmol) followed by
palladium acetate (0.084 g, 0.375 mmol) were added, and the
suspension was refluxed overnight. The solid was filtered, the
reaction mixture was evaporated. The black oil was purified by
flush column chromatography on silica gel (5% EtOAc:DCM). The
combined fractions were evaporated. The resulting oil was dissolved
in dioxane (20 mL) and sodium hydroxide (20 mL, 20.00 mmol) was
added and the reaction mixture was refluxed overnight. The layers
were separated and the organic layer was washed with 20 ml of 1 M
NaOH. The aqueous layers were combined and washed with EtOAc. The
combined organic layers were washed with water, brine and dried
over MgSO.sub.4 and filtered. The solution was evaporated,
suspended in acetonitrile and filtered.
4-Chloro-2-[(5-chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-
-5-yl]amino}-4-pyridinyl)amino]benzoic acid (260 mg, 0.619 mmol,
6.60% yield) was isolated as a yellow solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) ppm 1.28 (d, J=6.57 Hz, 6H) 1.91 (s, 1H) 2.13 (s, 3H)
4.43 (quin, J=6.57 Hz, 1H) 5.97 (s, 1H) 6.78 (s, 1H) 7.05 (dd,
J=8.46, 1.89 Hz, 1H) 7.48 (d, J=1.77 Hz, 1H) 7.96 (d, J=8.34 Hz,
1H) 8.03 (s, 1H) 8.61 (s, 1H); HPLC Rt=2.70 min, MS (ESI): 420.1,
422.0 [M+H].sup.+.
Step 3.
[0285] To the solution of
4-chloro-2-[(5-chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amin-
o}-4-pyridinyl)amino]benzoic acid (260 mg, 0.619 mmol) in
N,N-dimethylformamide (DMF) (20 mL) was added HOBT (114 mg, 0.742
mmol) and EDC (142 mg, 0.742 mmol) and the reaction mixture was
stirred for 30 min. To this solution was added O-methoxylamine
hydrochloride (62.0 mg, 0.742 mmol), and after 30 min the reaction
was cooled to 0.degree. C. and DIEA (0.323 mL, 1.856 mmol) was
added. The reaction mixture was stirred at the room temperature
over the weekend. Water (100 mL) followed by acetic acid (1 mL)
were added and the reaction mixture was extracted with 2.times.50
ml of ethyl acetate. The organic layer was washed with 2.times.50
ml sat KHCO.sub.3, brine, dried over MgSO.sub.4 and evaporated. The
resulting yellow oil was purified by flash column chromatography on
silica gel using DCM:EtOAc (10% to 100%).
4-Chloro-2-[(5-chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amin-
o}-4-pyridinyl)amino]-N-(methyloxy)benzamide (85 mg, 0.180 mmol,
29.1% yield) was isolated as white foam. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) ppm 1.27 (d, J=6.57 Hz, 6H) 2.12 (s, 3H) 3.70 (s, 3H)
4.41 (quin, J=6.57 Hz, 1H) 5.95 (s, 1H) 6.66 (s, 1H) 7.01-7.39 (m,
1H) 7.55-7.88 (m, 2H) 8.02 (s, 1H) 8.60 (s, 1H) 9.76 (br. s., 1H)
12.01 (br. s., 1H); HPLC Rt=2.50 min, MS (ESI): 449.0, 451.1
[M+H].sup.+.
Example 70
4-Chloro-2-[(5-chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino-
}-4-pyridinyl)amino]-N-(methyloxy)benzamide
##STR00137##
[0286] Step 1:
[0287] To a degassed solution of 2,5-dichloro-4-iodopyridine (4.5
g, 16.43 mmol), 2-amino-5-chlorobenzonitrile (2.507 g, 16.43 mmol)
and potassium triphosphate (10.46 g, 49.3 mmol) in 1,4-dioxane (60
ml) stirred under nitrogen at the room temperature was added
DPEPhos (0.708 g, 1.314 mmol) and palladium acetate (0.148 g, 0.657
mmol). The reaction mixture was stirred at reflux for 18 hr. The
reaction mixture was filtered. The reaction mixture was evaporated.
Ether (50 ml) was added and the solid was filtered.
5-chloro-2-[(2,5-dichloro-4-pyridinyl)amino]benzonitrile (1.8 g,
5.43 mmol, 33.0% yield) was isolated as orange solid. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) ppm 6.63 (s, 1H) 7.52 (d, J=8.59 Hz, 1H)
7.81 (dd, J=8.59, 2.53 Hz, 1H) 8.09 (d, J=2.53 Hz, 1H) 8.24 (s, 1H)
9.06 (br. s., 1H); HPLC Rt=3.53 min, MS (ESI): 298.0, 299.9
[M+H].sup.+.
Step 2.
[0288] To a solution of
5-chloro-2-[(2,5-dichloro-4-pyridinyl)amino]benzonitrile (1.8 g,
6.03 mmol) and 3-methyl-1-(1-methylethyl)-1H-pyrazol-5-amine (0.839
g, 6.03 mmol) in 1,4-dioxane (40 mL) was added cesium carbonate
(5.89 g, 18.09 mmol) and the reaction mixture was degassed. DPEPhos
(0.260 g, 0.482 mmol) was added followed by palladium acetate
(0.054 g, 0.241 mmol) and the reaction mixture was heated to reflux
overnight. The suspension was then filtered. The dioxane was
evaporated. Solid was partitioned between 1 M HCl and ethyl
acetate. Layers were separated, and organic layer discarded. The
HCl-containing layer was neutralized and extracted with 2.times.50
mL of ethyl acetate. Organic layers were combined, washed with
brine, dried over MgSO.sub.4, filtered and evaporated.
5-Chloro-2-[(5-chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amin-
o}-4-pyridinyl)amino]benzonitrile (850 mg, 2.118 mmol, 35.1% yield)
was isolated as a yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
ppm 1.25 (d, J=6.57 Hz, 6H) 2.09 (s, 3H) 4.35 (quin, J=6.57 Hz, 1H)
5.89 (s, 1H) 6.03 (s, 1H) 7.46 (d, J=8.84 Hz, 1H) 7.81 (dd, J=8.72,
2.65 Hz, 1H) 7.96 (s, 1H) 8.11 (d, J=2.53 Hz, 1H) 8.42 (s, 1H) 8.54
(s, 1H) HPLC Rt=2.60 min, MS (ESI): 400.8, 403.1 [M+H].sup.+.
Step 3.
[0289] A solution of
5-chloro-2-[(5-chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amin-
o}-4-pyridinyl)amino]benzonitrile (850 mg, 2.118 mmol) in sodium
hydroxide -1 M (20 mL, 20.00 mmol) and 1,4-dioxane (20 mL) was
refluxed overnight. Ethyl acetate was added and the layers were
separated. The organic layer was washed with 1 M NaOH (40 ml).
Combined aqueous layers were washed with ethyl acetate. The organic
layers were combined, evaporated, dissolved in MeOH and evaporated
again.
5-Chloro-2-[(5-chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amin-
o}-4-pyridinyl)amino]benzoic acid (800 mg, 1.903 mmol, 90% yield)
was isolated as a yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
ppm 1.28 (d, J=6.57 Hz, 6H) 2.12 (s, 3H) 4.40 (quin, J=6.51 Hz, 1H)
5.97 (s, 1H) 6.79 (s, 1H) 7.56-7.62 (m, 1H) 7.62-7.69 (m, 1H) 7.94
(d, J=2.27 Hz, 1H) 8.04 (s, 1H) 8.58 (s, 1H) 9.94 (br. s., 1H)
13.97 (br. s., 1H); HPLC Rt=2.65 min, MS (ESI): 420.2, 421.1
[M+H].sup.+.
Step 4.
[0290] To a solution of
5-chloro-2-[(5-chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amin-
o}-4-pyridinyl)amino]benzoic acid (830 mg, 1.975 mmol) in
N,N-dimethylformamide (20 mL) was added HOBT (363 mg, 2.370 mmol)
and EDC (454 mg, 2.370 mmol) and the reaction mixture was stirred
for 30 min. To this solution was added o-methoxylamine
hydrochloride (198 mg, 2.370 mmol) and after 30 min the mixture was
cooled to 0.degree. C. DIEA (1.032 mL, 5.92 mmol) was added. The
reaction mixture was stirred at the room temperature over the
weekend. Water (100 mL) was added followed by acetic acid (1 mL)
and the solution extracted with 2.times.50 ml of ethyl acetate. The
organic layer was washed with 2.times.50 ml sat KHCO.sub.3, brine,
dried over MgSO.sub.4 and evaporated. The product was purified by
flash column chromatography on silica gel using EtOAc:DCM (10% to
100%).
5-Chloro-2-[(5-chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amin-
o}-4-pyridinyl)amino]-N-(methyloxy)benzamide (250 mg, 0.529 mmol,
26.8% yield) was isolated as a white foam. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) ppm 1.27 (d, J=6.57 Hz, 6H) 2.11 (s, 3H) 3.71 (s, 3H)
4.20-4.55 (m, 1H) 5.94 (s, 1H) 6.63 (s, 1H) 7.59 (s, 2H) 7.67 (s,
1H) 7.99 (s, 1H) 8.50 (s, 1H) 9.43 (br. s., 1H) 12.03 (br. s., 1H);
HPLC Rt=2.46 min, MS (ESI): 449.1, 451.1 [M+H].sup.+.
Example 71
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyrid-
inyl)amino]-5-fluoro-N-(methyloxy)benzamide
##STR00138##
[0291] Step 1:
[0292] To a degassed solution of 2,5-dichloro-4-iodopyridine (8 g,
29.2 mmol), 2-amino-5-fluorobenzonitrile (3.98 g, 29.2 mmol) and
potassium triphosphate (18.60 g, 88 mmol) in 1,4-dioxane (100 ml)
stirred under nitrogen at the room temperature was added DPEPhos
(1.258 g, 2.337 mmol) and palladium acetate (0.262 g, 1.168 mmol).
The reaction mixture was stirred at the reflux for 18 hr. The
reaction mixture was filtered. The solvent was evaporated. Ether
(50 ml) was added and the solid was filtered.
2-[(2,5-Dichloro-4-pyridinyl)amino]-5-fluorobenzonitrile (7.09 g,
25.1 mmol, 86% yield) was isolated as an orange solid. .sup.1H NMR
(400 MHz, METHANOL-d.sub.4) ppm 6.44 (s, 1H) 7.46-7.58 (m, 2H) 7.66
(dd, J=8.08, 2.78 Hz, 1H) 8.08 (s, 1H); HPLC Rt=3.23 min, MS (ESI):
382.0, 384.19 [M+H].sup.+.
Step 2.
[0293] To a solution of
2-[(2,5-dichloro-4-pyridinyl)amino]-5-fluorobenzonitrile (7.09 g,
25.1 mmol) and 3-methyl-1-(1-methylethyl)-1H-pyrazol-5-amine (3.5
g, 25.1 mmol) in 1,4-dioxane (100 mL) was added cesium carbonate
(24.58 g, 75 mmol) and the reaction mixture was degassed. DPEPhos
(1.083 g, 2.012 mmol) was added followed by palladium acetate
(0.226 g, 1.006 mmol) and the reaction mixture was heated to reflux
overnight. The suspension was then filtered. The dioxane was
evaporated off. The solid was purified by flash column
chromatography on silica gel (10% DCM:EtOAC). Fractions were
collected and evaporated. The yellow oil was dissolved in diethyl
ether, sonicated and filtered.
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-5-fluorobenzonitrile (1.3 g, 3.21 mmol, 12.76% yield)
was isolated as a white solid; .sup.1H NMR (400 MHz,
METHANOL-d.sub.4) ppm 1.35 (d, J=6.82 Hz, 6H) 2.20 (s, 3H) 4.43
(quin, J=6.69 Hz, 1H) 5.82 (s, 1H) 5.89 (s, 1H) 7.47-7.57 (m, 2H)
7.61-7.74 (m, 1H) 7.88 (s, 1H); HPLC Rt=2.40 min, MS (ESI): 385.2
[M+H].sup.+.
Step 3.
[0294] A solution of
2-[(5-chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-5-fluorobenzonitrile (1.3 g, 3.38 mmol) in sodium
hydroxide -1 M (10 mL, 10.00 mmol) and 1,4-dioxane (10 mL) was
refluxed overnight. Ethyl acetate was added and the layers were
separated. The organic layer was washed with 1 M NaOH (40 ml). The
combined aqueous layers were washed with ethyl acetate, and
neutralized with acetic acid. The product was isolated by
filtration.
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-5-fluorobenzoic acid (1.1 g, 2.72 mmol, 27.2% yield)
was isolated as a yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
ppm 1.27 (d, J=6.57 Hz, 6H) 2.11 (s, 3H) 4.39 (quin, J=6.57 Hz, 1H)
5.95 (s, 1H) 6.70 (s, 1H) 7.51 (td, J=8.40, 3.16 Hz, 1H) 7.61 (dd,
J=9.09, 4.80 Hz, 1H) 7.72 (dd, J=9.22, 3.16 Hz, 1H) 8.00 (s, 1H)
8.53 (s, 1H) 9.66 (br. s., 1H) 13.88 (br. s., 1H); HPLC Rt=2.44
min, MS (ESI): 404.3 [M+H].sup.+.
Step 4.
[0295] To the solution of
2-[(5-chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-5-fluorobenzoic acid (1128 mg, 2.79 mmol) in
N,N-dimethylformamide (DMF) (20 mL) was added HOBT (513 mg, 3.35
mmol) followed by EDC (643 mg, 3.35 mmol) and the reaction mixture
stirred for 30 min. To this solution was added o-methoxylamine
hydrochloride (280 mg, 3.35 mmol) and after 30 min, at 0.degree. C.
DIEA (1.460 mL, 8.38 mmol) was added. The reaction mixture was
stirred at room temperature 24 hr. Water (100 mL) was added
followed by acetic acid (1 mL) and the solution extracted with
2.times.50 ml of ethyl acetate. The organic layer was separated,
washed with 2.times.50 ml sat KHCO.sub.3, brine, dried over
MgSO.sub.4 and evaporated. The resulting oil was suspended in
dichloromethane and filtered.
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-5-fluoro-N-(methyloxy)benzamide (620 mg, 1.361 mmol,
48.7% yield) was isolated as a white solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) ppm 1.26 (d, J=6.57 Hz, 6H) 2.10 (s, 3H) 3.69 (s, 3H)
4.38 (quin, J=6.57 Hz, 1H) 5.92 (s, 1H) 6.50 (s, 1H) 7.41-7.52 (m,
2H) 7.58 (dd, J=8.72, 4.67 Hz, 1H) 7.96 (s, 1H) 8.46 (s, 1H) 9.10
(s, 1H) 11.95 (s, 1H); HPLC Rt=2.24 min, MS (ESI): 433.4
[M+H].sup.+.
Example 72
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyrid-
inyl)amino]-3-fluoro-N-(methyloxy)benzamide
##STR00139##
[0296] Step 1:
[0297] To a degassed solution of 2,5-dichloro-4-iodopyridine (8 g,
29.2 mmol), 2-amino-3-fluorobenzonitrile (3.98 g, 29.2 mmol) and
potassium triphosphate (18.60 g, 88 mmol) in 1,4-dioxane (60 ml)
stirred under nitrogen at the room temperature was added DPEPhos
(1.258 g, 2.337 mmol) and palladium acetate (0.262 g, 1.168 mmol)
The reaction mixture was stirred at reflux for 18 hr. The reaction
mixture was filtered. 3-Methyl-1-(1-methylethyl)-1H-pyrazol-5-amine
(4.07 g, 29.2 mmol) and cesium carbonate (28.6 g, 88 mmol) were
added. The reaction mixture was degassed and palladium acetate
(0.262 g, 1.168 mmol) and DPEPhos (1.258 g, 2.337 mmol) were added.
The reaction mixture was refluxed overnight. The reaction mixture
was filtered and the solid was dissolved in water, heated to
50.degree. C. and stirred for 10 minutes, then filtered again.
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-3-fluorobenzonitrile (6 g, 15.59 mmol, 53.4% yield)
was isolated as an orange solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) d ppm 1.23 (d, J=6.57 Hz, 6H) 2.07 (s, 3H) 4.32
(quin, J=6.57 Hz, 1H) 5.64 (d, J=2.02 Hz, 1H) 5.83 (s, 1H)
7.39-7.58 (m, 1H) 7.63-7.82 (m, 2H) 7.88 (s, 1H) 8.26 (br. s., 1H)
8.41 (br. s., 1H); HPLC Rt=2.35 min, MS (ESI): 385.0
[M+H].sup.+.
Step 2.
[0298] A solution of
2-[(5-chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-3-fluorobenzonitrile (4.5 g, 11.69 mmol) in sodium
hydroxide -1 M (10 mL, 10.00 mmol) and 1,4-dioxane (10 mL) was
refluxed overnight. Ethyl acetate was added and the layers were
separated. The organic layer was washed with 1 M NaOH (40 ml).
Combined aqueous layers were washed with ethyl acetate, and
neutralized with acetic acid. The solid was filtered.
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]ami-
no}-4-pyridinyl)amino]-3-fluorobenzoic acid (3.2 g, 7.53 mmol, 75%
yield) was isolated as a yellow solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) ppm 1.24 (d, J=6.57 Hz, 6H) 2.09 (s, 3H) 4.34 (quin,
J=6.57 Hz, 1H) 5.86 (s, 1H) 5.91 (d, J=5.81 Hz, 1H) 7.35 (td,
J=8.02, 4.93 Hz, 1H) 7.54-7.65 (m, 1H) 7.81 (d, J=7.58 Hz, 1H) 7.94
(s, 1H) 8.45 (s, 1H) 8.88 (br. s., 1H) 13.74 (br. s., 1H); HPLC
Rt=2.36 min, MS (ESI): 404.3 [M+H].sup.+.
Step 3.
[0299] To a solution of
2-[(5-chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-3-fluorobenzoic acid (3.2 g, 7.92 mmol) in
N,N-dimethylformamide (DMF) (50 mL) was added HOBT (1.456 g, 9.51
mmol) and EDC (1.823 g, 9.51 mmol) and the reaction mixture was
stirred for 30 min. To this solution was added O-methoxylamine
hydrochloride (0.794 g, 9.51 mmol). After 30 min the reaction
mixture was cooled to 0.degree. C. and DIEA (4.14 mL, 23.77 mmol)
was added. The reaction mixture was stirred at the room temperature
24 hr. Water (100 mL) was added followed by acetic acid (1 mL) and
the solution was extracted with 2.times.50 ml of ethyl acetate. The
combined organic layers were washed with 2.times.50 ml sat
KHCO.sub.3, brine, dried over MgSO.sub.4 and evaporated. The oil
was suspended in dichloromethane and filtered.
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-3-fluoro-N-(methyloxy)benzamide (1.1 g, 2.414 mmol,
30.5% yield) was isolated as a white solid; .sup.1H NMR (400 MHz,
DMSO-d.sub.6) ppm 1.24 (d, J=6.57 Hz, 6H) 2.08 (s, 3H) 3.68 (s, 3H)
4.33 (quin, J=6.57 Hz, 1H) 5.83 (d, J=5.05 Hz, 1H) 5.84 (s, 1H)
7.30-7.49 (m, 2H) 7.49-7.63 (m, 1H) 7.92 (s, 1H) 8.41 (d, J=4.29
Hz, 2H) 11.85 (s, 1H); HPLC Rt=2.18 min, MS (ESI): 433.3
[M+H].sup.+.
Example 73
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyrid-
inyl)amino]-4-fluoro-N-(methyloxy)benzamide
##STR00140##
[0300] Step 1:
[0301] To a degassed solution of 2,5-dichloro-4-iodopyridine (5 g,
18.26 mmol), 2-amino-4-fluorobenzonitrile (2.485 g, 18.26 mmol) and
potassium triphosphate (11.63 g, 54.8 mmol) in 1,4-dioxane (60 ml)
stirred under nitrogen at the room temperature was added DPEPhos
(0.787 g, 1.460 mmol) and palladium acetate (0.164 g, 0.730 mmol).
The reaction mixture was stirred at the reflux for 18 hr. The
reaction mixture was filtered.
3-Methyl-1-(1-methylethyl)-1H-pyrazol-5-amine (2.54 g, 18.26 mmol)
and cesium carbonate (17.84 g, 54.8 mmol) were added. The reaction
mixture was degassed and palladium acetate (0.164 g, 0.730 mmol)
and DPEPhos (0.787 g, 1.460 mmol) were added. The reaction mixture
was refluxed overnight. The reaction mixture was filtered. NaOH (60
mL, 60.0 mmol) was added and the reaction mixture refluxed
overnight. Ethyl acetate was added and the layers were separated.
The combined organics were washed with 1 M NaOH (40 ml). The
combined aqueous layers were washed with ethyl acetate, and
neutralized with acetic acid.
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-4-fluorobenzoic acid (2.5 g, 6.19 mmol, 33.9% yield)
was isolated by filtration as a yellow solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) ppm 1.29 (d, J=6.57 Hz, 6H) 2.12 (s, 3H) 4.43 (quin,
J=6.57 Hz, 1H) 5.99 (s, 1H) 6.86 (s, 1H) 6.87-6.93 (m, 1H) 7.34
(dd, J=11.62, 2.53 Hz, 1H) 8.03-8.10 (m, 2H) 8.62 (s, 1H) 10.65
(br. s., 1H); HPLC Rt=2.57 min, MS (ESI): 404.2 [M+H].sup.+.
Step 2.
[0302] To a solution of
2-[(5-chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-4-fluorobenzoic acid (2.5 g, 6.19 mmol) in
N,N-dimethylformamide (DMF) (50 mL) was added HOBT (1.138 g, 7.43
mmol) and EDC (1.424 g, 7.43 mmol) and the reaction mixture was
stirred for 30 min. To this solution was added O-methoxylamine
hydrochloride (0.620 g, 7.43 mmol) and after 30 min the mixture was
cooled to 0.degree. C. Then DIEA (3.23 mL, 18.57 mmol) was added.
The reaction mixture was stirred at the room temperature for 24 hr.
Water (100 mL) followed by acetic acid (1 mL) were added and the
solution extracted with 2.times.50 ml of ethyl acetate. The organic
layer was washed with 2.times.50 ml sat KHCO.sub.3, brine, dried
over MgSO.sub.4 and condensed. The resulting oil was purified by
flash column chromatography on silica gel (2:1 DCM:EtOAc).
2-[(5-Chloro-2-{[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-4-fluoro-N-(methyloxy)benzamide (1 g, 2.195 mmol,
35.5% yield) was isolated as a yellow foam. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) ppm 1.28 (d, J=6.57 Hz, 6H) 2.11 (s, 3H) 3.71 (s, 3H)
4.41 (quin, J=6.51 Hz, 1H) 5.97 (s, 1H) 6.74 (s, 1H) 6.87-7.06 (m,
1H) 7.39 (dd, J=11.37, 2.53 Hz, 1H) 7.66 (dd, J=8.46, 6.69 Hz, 1H)
8.03 (s, 1H) 8.57 (s, 1H) 9.96 (br. s., 1H) 11.98 (br. s., 1H);
HPLC Rt=2.36 min, MS (ESI): 433.3 [M+H].sup.+.
Example 74
[0303] Following substantially the procedure of Example 8 the
following compounds can be made starting with either
2,5-dichloro-4-iodopyridine or
2-chloro-4-iodo-5-(trifluoromethyl)pyridine and the appropriately
substituted 5-aminopyrazole.
74(a).
2-({5-Chloro-2-[(1,3-dimethyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}am-
ino)-3-fluoro-N-methylbenzamide
##STR00141##
[0304] 74(b).
2-({5-Chloro-2-[(1-ethyl-3-methyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amin-
o)-3-fluoro-N-methylbenzamide
##STR00142##
[0305] 74(c).
2-({5-Chloro-2-[(1,5-dimethyl-1H-pyrazol-4-yl)amino]-4-pyridinyl}amino)-3-
-fluoro-N-methylbenzamide
##STR00143##
[0306] 74(d).
2-{[2-[(1-Ethyl-3-methyl-1H-pyrazol-5-yl)amino]-5-(trifluoromethyl)-4-pyr-
idinyl]amino}-3-fluoro-N-methylbenzamide
##STR00144##
[0307] 74(e).
2-{[2-[(1,3-Dimethyl-1H-pyrazol-5-yl)amino]-5-(trifluoromethyl)-4-pyridin-
yl]amino}-3-fluoro-N-methylbenzamide
##STR00145##
[0308] 74(f).
2-{[2-{[1-Ethyl-3-(hydroxymethyl)-1H-pyrazol-5-yl]amino}-5-chloro-4-pyrid-
inyl]amino}-3-fluoro-N-methylbenzamide
##STR00146##
[0309] 74(g).
3-Fluoro-2-{[2-{[3-(hydroxymethyl)-1-methyl-1H-pyrazol-5-yl]amino}-5-chlo-
ro-4-pyridinyl]amino}-N-methylbenzamide
##STR00147##
[0310] 74(h).
2-{[2-{[1-Ethyl-3-(2-hydroxyethyl)-1H-pyrazol-5-yl]amino}-5-chloro-4-pyri-
dinyl]amino}-3-fluoro-N-methylbenzamide
##STR00148##
[0311] 74(i).
3-Fluoro-2-{[2-{[3-(2-hydroxyethyl)-1-methyl-1H-pyrazol-5-yl]amino}-5-chl-
oro-4-pyridinyl]amino}-N-methylbenzamide
##STR00149##
[0312] 74(j).
2-{[2-({3-[(Dimethylamino)methyl]-1-methyl-1H-pyrazol-5-yl}amino)-5
chloro-4-pyridinyl]amino}-3-fluoro-N-methylbenzamide
##STR00150##
[0313] 74(k).
2-{[2-({3-[(Dimethylamino)methyl]-1-ethyl-1H-pyrazol-5-yl}amino)-5
chloro-4-pyridinyl]amino}-3-fluoro-N-methylbenzamide
##STR00151##
[0314] 74(l).
2-({5-Chloro-2-[(3-{[ethyl(methyl)amino]methyl}-1-methyl-1H-pyrazol-5-yl)-
amino]-4-pyridinyl}amino)-3-fluoro-N-methylbenzamide
##STR00152##
[0315] 74(m).
2-{[5-chloro-2-({3-[(diethylamino)methyl]-1-methyl-1H-pyrazol-5-yl}amino)-
-4-pyridinyl]amino}-3-fluoro-N-methylbenzamide
##STR00153##
[0317] 74(n).
2-{[2-{[1-Ethyl-3-(hydroxymethyl)-1H-pyrazol-5-yl]amino}-5-(trifluorometh-
yl)-4-pyridinyl]amino}-3-fluoro-N-methylbenzamide
##STR00154##
74(o).
3-Fluoro-2-{[2-{[3-(hydroxymethyl)-1-methyl-1H-pyrazol-5-yl]amino}-
-5-(trifluoromethyl)-4-pyridinyl]amino}-N-methylbenzamide
##STR00155##
[0318] 74(p).
2-{[2-{[1-Ethyl-3-(2-hydroxyethyl)-1H-pyrazol-5-yl]amino}-5-(trifluoromet-
hyl)-4-pyridinyl]amino}-3-fluoro-N-methylbenzamide
##STR00156##
[0319] 74(q).
3-Fluoro-2-{[2-{[3-(2-hydroxyethyl)-1-methyl-1H-pyrazol-5-yl]amino}-5-(tr-
ifluoromethyl)-4-pyridinyl]amino}-N-methylbenzamide
##STR00157##
[0320] 74(r).
2-{[2-({3-[(Dimethylamino)methyl]-1-methyl-1H-pyrazol-5-yl}amino)-5-(trif-
luoromethyl)-4-pyridinyl]amino}-3-fluoro-N-methylbenzamide
##STR00158##
[0321] Example 75
[0322] Following substantially the procedure of Example 72 the
following compounds can be made using the appropriately substituted
5-aminopyrazole.
75(a).
2-({5-Chloro-2-[(1-ethyl-3-methyl-1H-pyrazol-5-yl)amino]-4-pyridiny-
l}amino)-3-fluoro-N-(methyloxy)benzamide
##STR00159##
[0323] 75(b).
2-({5-Chloro-2-[(1,3-dimethyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amino)-3-
-fluoro-N-(methyloxy)benzamide
##STR00160##
[0324] 75(c).
2-[(5-Chloro-2-{[1-ethyl-3-(hydroxymethyl)-1H-pyrazol-5-yl]amino}-4-pyrid-
inyl)amino]-3-fluoro-N-(methyloxy)benzamide
##STR00161##
[0325] 75(d).
2-[(5-Chloro-2-{[3-(hydroxymethyl)-1-methyl-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-3-fluoro-N-(methyloxy)benzamide
##STR00162##
[0326] 75(e).
2-{[5-Chloro-2-({3-[(dimethylamino)methyl]-1-ethyl-1H-pyrazol-5-yl}amino)-
-4-pyridinyl]amino}-3-fluoro-N-(methyloxy)benzamide
##STR00163##
[0327] 75(f).
2-({5-Chloro-2-[(1,5-dimethyl-1H-pyrazol-4-yl)amino]-4-pyridinyl}amino)-3-
-fluoro-N-(methyloxy)benzamide
##STR00164##
[0328] Example 76
[0329] Following substantially the procedure of Example 41a or 41b
the following compound can be made using
3-[(dimethylamino)methyl]-1-ethyl-1H-pyrazol-5-amine.
2-{[5-Chloro-2-({3-[(dimethylamino)methyl]-1-ethyl-1H-pyrazol-5-yl}amino)--
4-pyridinyl]amino}-N-(methyloxy)benzamide
##STR00165##
[0330] Example 77
[0331] Following substantially the procedure of Example 73 the
following compounds can be made using the appropriately substituted
5-aminopyrazole.
77(a).
2-({5-Chloro-2-[(1-ethyl-3-methyl-1H-pyrazol-5-yl)amino]-4-pyridiny-
l}amino)-4-fluoro-N-(methyloxy)benzamide
##STR00166##
[0332] 77(b).
2-({5-Chloro-2-[(1,3-dimethyl-1H-pyrazol-5-yl)amino]-4-pyridinyl}amino)-4-
-fluoro-N-(methyloxy)benzamide
##STR00167##
[0333] 77(c).
2-[(5-Chloro-2-{[1-ethyl-3-(hydroxymethyl)-1H-pyrazol-5-yl]amino}-4-pyrid-
inyl)amino]-4-fluoro-N-(methyloxy)benzamide
##STR00168##
[0334] 77(d).
2-[(5-Chloro-2-{[1-ethyl-3-(2-hydroxyethyl)-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-4-fluoro-N-(methyloxy)benzamide
##STR00169##
[0335] 77(e).
2-[(5-Chloro-2-{[3-(hydroxymethyl)-1-methyl-1H-pyrazol-5-yl]amino}-4-pyri-
dinyl)amino]-4-fluoro-N-(methyloxy)benzamide
##STR00170##
[0336] 77(f).
2-{[5-Chloro-2-({3-[(dimethylamino)methyl]-1-ethyl-1H-pyrazol-5-yl}amino)-
-4-pyridinyl]amino}-4-fluoro-N-(methyloxy)benzamide
##STR00171##
[0337] 77(g).
2-({5-Chloro-2-[(1,5-dimethyl-1H-pyrazol-4-yl)amino]-4-pyridinyl}amino)-4-
-fluoro-N-(methyloxy)benzamide
##STR00172##
Sequence CWU 1
1
2117PRTArtificial SequencePeptide substrate 1Arg Arg Arg Arg Arg
Arg Ser Glu Thr Asp Asp Tyr Ala Glu Ile Ile1 5 10 15
Asp214PRTArtificial SequencePeptide substrate 2Cys Ser Glu Thr Asp
Asp Tyr Ala Glu Ile Ile Asp Cys His1 5 10
* * * * *
References