U.S. patent application number 13/098243 was filed with the patent office on 2012-03-08 for cyclopropyl dicarboxamides and analogs exhibiting anti-cancer and anti-proliferative activities.
This patent application is currently assigned to DECIPHERA PHARMACEUTICALS, LLC. Invention is credited to DANIEL L. FLYNN, MICHAEL D. KAUFMAN.
Application Number | 20120058985 13/098243 |
Document ID | / |
Family ID | 44260801 |
Filed Date | 2012-03-08 |
United States Patent
Application |
20120058985 |
Kind Code |
A1 |
FLYNN; DANIEL L. ; et
al. |
March 8, 2012 |
CYCLOPROPYL DICARBOXAMIDES AND ANALOGS EXHIBITING ANTI-CANCER AND
ANTI-PROLIFERATIVE ACTIVITIES
Abstract
The disclosed compounds are useful in the treatment of mammalian
cancers and especially human cancers. Compounds, pharmaceutical
compositions, and methods of Formula I are disclosed: ##STR00001##
or a pharmaceutically acceptable salt, hydrate, solvate,
enantiomer, stereoisomer, or tautomer thereof.
Inventors: |
FLYNN; DANIEL L.; (LAWRENCE,
KS) ; KAUFMAN; MICHAEL D.; (LAWRENCE, KS) |
Assignee: |
DECIPHERA PHARMACEUTICALS,
LLC
LAWRENCE
KS
|
Family ID: |
44260801 |
Appl. No.: |
13/098243 |
Filed: |
April 29, 2011 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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61329548 |
Apr 29, 2010 |
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Current U.S.
Class: |
514/210.18 ;
514/245; 514/253.01; 514/269; 514/272; 514/349; 544/211; 544/319;
544/321; 544/364; 546/268.1; 546/297 |
Current CPC
Class: |
A61P 25/00 20180101;
A61P 11/06 20180101; A61P 35/02 20180101; C07D 401/12 20130101;
A61P 35/04 20180101; A61P 11/00 20180101; A61P 27/02 20180101; A61K
31/44 20130101; C07D 239/47 20130101; A61P 25/28 20180101; A61P
35/00 20180101; A61P 3/00 20180101; C07D 213/75 20130101; A61K
31/505 20130101; A61P 9/10 20180101; A61P 19/02 20180101; A61P
43/00 20180101; A61P 29/00 20180101 |
Class at
Publication: |
514/210.18 ;
514/245; 514/253.01; 514/269; 514/272; 514/349; 544/211; 544/319;
544/321; 544/364; 546/268.1; 546/297 |
International
Class: |
A61K 31/4427 20060101
A61K031/4427; A61K 31/496 20060101 A61K031/496; A61K 31/505
20060101 A61K031/505; A61K 31/44 20060101 A61K031/44; C07D 251/42
20060101 C07D251/42; C07D 239/47 20060101 C07D239/47; C07D 401/14
20060101 C07D401/14; C07D 401/12 20060101 C07D401/12; C07D 213/72
20060101 C07D213/72; A61P 11/06 20060101 A61P011/06; A61P 35/00
20060101 A61P035/00; A61P 25/00 20060101 A61P025/00; A61P 25/28
20060101 A61P025/28; A61P 3/00 20060101 A61P003/00; A61P 35/04
20060101 A61P035/04; A61P 35/02 20060101 A61P035/02; A61P 27/02
20060101 A61P027/02; A61P 19/02 20060101 A61P019/02; A61K 31/53
20060101 A61K031/53 |
Claims
1. A compound of Formula I, ##STR00030## or a pharmaceutically
acceptable salt, hydrate, solvate, enantiomer, stereoisomer, or
tautomer thereof, wherein X is halogen; Z1 and Z2 are independently
and individually CR2 or N; Z3 is CH or N; with the proviso that
ring B is not a tetrazine; each R1 is independently and
individually halogen, H, C1-C6 alkyl, branched C3-C7 alkyl, C3-C7
cycloalkyl, or --CN; each R2 is individually and independently H,
halogen, C1-C6 alkyl, or cyano; R3 is --C(O)R4,
--C(O)--C6-C10-aryl, --C(O)--C4-C6-heterocyclyl, or
--C(O)--C5-C6-heteroaryl, wherein aryl is phenyl, naphthyl,
tetrahydronaphthyl, indenyl or indanyl; and with the proviso that
when R3 is --C(O)--C4-C6-heterocyclyl, the heterocyclyl does not
have a N bonding hand to --C(O); R4 is C1-C7 alkyl, C3-C8
cycloalkyl, --(CH.sub.2).sub.p--CN, --(CH.sub.2).sub.p--OR6,
--(CH.sub.2).sub.p--NR6(R7),
--(CH.sub.2).sub.p--SO.sub.2--C1-C6-alkyl,
--(CH.sub.2).sub.p--C6-C10-aryl,
--(CH.sub.2).sub.p--C5-C6-heteroaryl, or
--(CH.sub.2).sub.p--C4-C6-heterocyclyl, wherein each alkyl or
alkylene is optionally substituted with one or two C1-C6 alkyl; and
aryl is phenyl, naphthyl, tetrahydronaphthyl, indenyl or indanyl;
each R6 and R7 is individually and independently H, C1-C6 alkyl, or
C3-C8 branched alkyl; each cycloalkyl, aryl, heteroaryl and
heterocyclyl is independently substituted with --(R25).sub.m; each
R25 is individually and independently C1-C6 alkyl, branched C3-C8
alkyl, halogen, --(CH.sub.2).sub.m--CN, --(CH.sub.2).sub.m--OR6,
--(CH.sub.2).sub.m--NR6(R7),
--(CH.sub.2).sub.m--SO.sub.2--C1-C6-alkyl,
--(CH.sub.2).sub.m--C(O)NR6(R7),
--(CH.sub.2).sub.m--C(O)--C4-C6-heterocyclyl, or
--(CH.sub.2).sub.m--C4-C6-heterocyclyl, wherein each alkyl or
alkylene is optionally substituted with one or two C1-C6 alkyl;
each m is individually and independently 0, 1, 2, or 3; and p is 1,
2, or 3.
2. The compound of claim 1, wherein Z1 and Z2 are CR2, and Z3 is
CH.
3. The compound of claim 2, wherein the compound is a compound of
Formula Ic, ##STR00031## or a pharmaceutically acceptable salt,
hydrate, solvate, enantiomer, stereoisomer, or tautomer thereof,
and wherein n is 0, 1, or 2.
4. The compound of claim 3, wherein R3 is --C(O)R4.
5. The compound of claim 4, wherein R4 is C1-C7 alkyl, C3-C8
cycloalkyl, --(CH.sub.2).sub.p--CN, --(CH.sub.2).sub.p--OR6,
--(CH.sub.2).sub.p--NR6(R7),
--(CH.sub.2).sub.p--SO.sub.2--C1-C6-alkyl, or
--(CH.sub.2).sub.p--C4-C6-heterocyclyl, and wherein each alkyl or
alkylene is optionally substituted with one or two C1-C6 alkyl.
6. The compound of claim 4, wherein R4 is C1-C7 alkyl or C3-C8
cycloalkyl, and wherein each alkyl or alkylene is optionally
substituted with one or two C1-C6 alkyl.
7. The compound of claim 3, wherein R3 is --C(O)--C6-C10-aryl,
--C(O)--C4-C6-heterocyclyl, or --C(O)--C5-C6-heteroaryl.
8. The compound of claim 1, wherein Z1 and Z2 are CR2, and Z3 is
N.
9. The compound of claim 8, wherein the compound is a compound of
Formula If, ##STR00032## or a pharmaceutically acceptable salt,
hydrate, solvate, enantiomer, stereoisomer, or tautomer thereof,
and wherein n is 0, 1, or 2.
10. The compound of claim 9, wherein R3 is --C(O)R4.
11. The compound of claim 9, wherein R3 is --C(O)--C6-C10-aryl,
--C(O)--C4-C6-heterocyclyl, or --C(O)--C5-C6-heteroaryl.
12. The compound of claim 1, wherein Z1 is CR2, Z2 is N, and Z3 is
CH.
13. The compound of claim 12, wherein the compound is a compound of
Formula Ij, ##STR00033## or a pharmaceutically acceptable salt,
hydrate, solvate, enantiomer, stereoisomer, or tautomer
thereof.
14. The compound of claim 13, wherein R3 is --C(O)R4.
15. The compound of claim 13, wherein R3 is --C(O)--C6-C10-aryl,
--C(O)--C4-C6-heterocyclyl, or --C(O)--C5-C6-heteroaryl.
16. The compound of claim 1, wherein Z1 is CR2, and Z2 and Z3 are
N.
17. The compound of claim 16, wherein the compound is a compound of
Formula Im, ##STR00034## or a pharmaceutically acceptable salt,
hydrate, solvate, enantiomer, stereoisomer, or tautomer
thereof.
18. The compound of claim 17, wherein R3 is --C(O)R4.
19. The compound of claim 17, wherein R3 is --C(O)--C6-C10-aryl,
--C(O)--C4-C6-heterocyclyl, or --C(O)--C5-C6-heteroaryl.
20. A compound selected from the group consisting of
N-(4-(2-acetamidopyridin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluorophenyl)-
cyclopropane-1,1-dicarboxamide,
N-(4-(2-acetamidopyridin-4-yloxy)-5-chloro-2-fluorophenyl)-N-(4-fluorophe-
nyl)cyclopropane-1,1-dicarboxamide,
N-(4-(2-acetamidopyridin-4-yloxy)-2,5-difluorophenyl)-N-phenylcyclopropan-
e-1,1-dicarboxamide,
N-(4-(2-(2-(dimethylamino)acetamido)pyridin-4-yloxy)-2,5-difluorophenyl)--
N'-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
N-(4-(2-acetamidopyrimidin-4-yloxy)-2,5-difluorophenyl)-N-(4-fluorophenyl-
)cyclopropane-1,1-dicarboxamide,
N-(4-(2-(cyclopropanecarboxamido)pyridin-4-yloxy)-2,5-difluorophenyl)-N-(-
4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
N-(2,5-difluoro-4-(2-propionamidopyridin-4-yloxy)phenyl)-N'-(4-fluorophen-
yl)cyclopropane-1,1-dicarboxamide,
N-(2,5-difluoro-4-((2-(2-methoxyacetamido)pyridin-4-yl)oxy)phenyl)-N'-(4--
fluorophenyl)cyclopropane-1,1-dicarboxamide,
N-(2,5-difluoro-4-(2-isobutyramidopyridin-4-yloxy)phenyl)-N'-(4-fluorophe-
nyl)cyclopropane-1,1-dicarboxamide,
N-(4-(2-(2-cyanoacetamido)pyridin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluo-
rophenyl)cyclopropane-1,1-dicarboxamide,
N-(4-(2-(azetidine-3-carboxamido)pyridin-4-yloxy)-2,5-difluorophenyl)-N'--
(4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
N-(4-(2-(cyclobutanecarboxamido)pyridin-4-yloxy)-2,5-difluorophenyl)-N'-(-
4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
(R)--N-(2,5-difluoro-4-((2-(2-methoxypropanamido)pyridin-4-yl)oxy)phenyl)-
-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
(R)--N-(2,5-difluoro-4-((2-(2-hydroxypropanamido)pyridin-4-yl)oxy)phenyl)-
-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
N-(2,5-difluoro-4-((2-pivalamidopyridin-4-yl)oxy)phenyl)-N-(4-fluoropheny-
l)cyclopropane-1,1-dicarboxamide,
(S)--N-(2,5-difluoro-4-((2-(2-methoxypropanamido)pyridin-4-yl)oxy)phenyl)-
-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
(S)-1-((4-(2,5-difluoro-4-(1-((4-fluorophenyl)carbamoyl)cyclopropanecarbo-
xamido)phenoxy)pyridin-2-yl)amino)-1-oxopropan-2-yl acetate,
N-(2,5-difluoro-4-((2-(2-fluoro-2-methylpropanamido)pyridin-4-yl)oxy)phen-
yl)-N'-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide and
pharmaceutically acceptable salts, solvates, hydrates and tautomers
thereof.
21. A compound selected from
N-(4-(2-(cyclopropanecarboxamido)pyridin-4-yloxy)-2,5-difluorophenyl)-N'--
(4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
N-(2,5-difluoro-4-(2-propionamidopyridin-4-yloxy)phenyl)-N'-(4-fluorophen-
yl)cyclopropane-1,1-dicarboxamide,
N-(2,5-difluoro-4-(2-isobutyramidopyridin-4-yloxy)phenyl)-N'-(4-fluorophe-
nyl)cyclopropane-1,1-dicarboxamide,
N-(4-(2-acetamidopyridin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluorophenyl)-
cyclopropane-1,1-dicarboxamide.
22. A method of treating mammalian disease wherein the disease
etiology or progression is at least partially mediated by a kinase
activity, wherein the kinase is a wildtype form, a mutant oncogenic
form, an aberrant fusion protein form or a polymorph, the method
comprising administering to a mammal in need thereof an effective
amount of a compound of claim 1.
23. The method of claim 22, wherein the disease etiology or
progression is at least partially mediated by the kinase activity
of c-MET, mutant oncogenic forms, aberrant fusion proteins, or
polymorphs thereof.
24. A pharmaceutical composition, comprising a compound of claim 1
and a pharmaceutically acceptable carrier.
25. The composition of claim 24, further comprising an additive
selected from adjuvants, excipients, diluents, or stabilizers.
26. A method of treating cancer, gastrointestinal stromal tumors,
hyperproliferative diseases, metabolic diseases, neurodegenerative
diseases, or diseases characterized by angiogenesis, such as solid
tumors, melanomas, glioblastomas, ovarian cancer, pancreatic
cancer, prostate cancer, lung cancers, breast cancers, renal
cancers, hepatic cancers, cervical carcinomas, metastasis of
primary tumor sites, myeloproliferative diseases, chronic
myelogenous leukemia, leukemias, papillary thyroid carcinoma,
non-small cell lung cancer, mesothelioma, hypereosinophilic
syndrome, colonic cancers, ocular diseases characterized by
hyperproliferation leading to blindness including retinopathies,
diabetic retinopathy, age-related macular degeneration,
hypereosinophilic syndrome, rheumatoid arthritis, asthma, chronic
obstructive pulmonary, mastocytosis, or mast cell leukemia, the
method comprising administering to a patient in need thereof an
effective amount of a compound of claim 1.
27. The method of claim 26, wherein the compound is administered
orally, parenterally, by inhalation, or subcutaneously.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional
Patent Application No. 61/329,548, filed Apr. 29, 2010, entitled
"CYCLOPROPYL DICARBOXAMIDES AND ANALOGS EXHIBITING ANTI-CANCER AND
ANTI-PROLIFERATIVE ACTIVITIES, which is incorporated herein by
reference in its entirety.
FIELD
[0002] The present invention relates to kinase inhibitors
exhibiting novel and unexpected properties useful for the treatment
of various diseases including hyperproliferative diseases and
cancer. More particularly, the invention is concerned with such
compounds, methods of treating diseases, and methods of synthesis
of the compounds. Preferably, the compounds are useful for the
modulation of activity of c-MET kinase, c-MET kinase polymorphs,
c-MET kinase mutants, or c-MET kinase fusion proteins in the
treatment of mammalian diseases, and in particular human
hyperproliferative diseases and human cancers. In some embodiments,
compounds disclosed herein exhibit unexpected selectivity for
modulation of c-MET kinase activity.
BACKGROUND OF THE INVENTION
[0003] c-MET is a receptor tyrosine kinase (RTK) located on
chromosome 7p and activated via its natural ligand hepatocyte
growth factor. c-MET is found mutated in a variety of solid tumors
(Ma, P. C. et al. Cancer Metastasis (2003) 22: 309). Mutations in
the tyrosine kinase domain are associated with hereditary papillary
renal cell carcinomas (Schmidt, L. et al. Nat. Genet. (1997) 16:
68; Schmidt, L. et al. Oncogene (1999) 18: 2343), whereas mutations
in the sema and juxtamembrane domains are often found in small cell
lung cancers (Ma, P. C. et al. Cancer Res. (2003) 63: 6272). Many
activating mutations are also found in breast cancers (Nakopoulou,
et al. Histopath. (2000) 36(4): 313). The panoply of tumor types
for which c-MET mediated growth has been implicated suggests this
is a target ideally suited for modulation by specific c-MET small
molecule inhibitors.
[0004] The TPR-MET oncogene is a transforming variant of the c-MET
RTK and was initially identified after treatment of a human
osteogenic sarcoma cell line transformed by the chemical carcinogen
N-methyl-N'-nitro-N-nitrosoguanidine (Park, M. et al. Cell (1986)
45: 895). The TPR-MET fusion oncoprotein is the result of a
chromosomal translocation, placing the TPR3 locus on chromosome 1
upstream of a portion of the c-MET gene on chromosome 7 encoding
only for the cytoplasmic region. Studies suggest that TPR-MET is
detectable in experimental cancers (e.g., Yu, J. et al. Cancer
(2000) 88: 1801). Dimerization of the M.sub.r 65,000 TPR-MET
oncoprotein through a leucine zipper motif encoded by TPR leads to
constitutive activation of the c-MET kinase (then, Z. et al.
Oncogene (1994) 9: 1691). TPR-MET activates wild-type c-MET RTK and
can activate crucial cellular growth pathways, including the Ras
pathway (Aklilu, F. et al. Arm. J. Physiol. (1996) 271: E277) and
the phosphatidylinositol 3-kinase (PI3K)/AKT pathway (Ponzetto, C.
et al. Mol. Cell. Biol. (1993) 13: 4600). Conversely, in contrast
to c-MET RTK, TPR-MET is ligand independent, lacks the CBL-like SH2
domain binding site in the juxtamembrane region in c-MET, and is
mainly cytoplasmic. c-MET immunohistochemical expression seems to
be associated with abnormal .beta.-catenin expression, a hallmark
feature of epithelial to mesenchymal transition (EMT) and provides
good prognostic and predictive factors in breast cancer
patients.
[0005] In human therapeutics, it is desirable to provide small
molecule inhibitors of a protein target within in a protein family
which do not cross-inhibit closely related protein family members.
These closely related protein family members are often referred to
as `off-targets`, to distinguish them from the essential target of
interest referred to as the `on target` of the inhibitor. A small
molecule which inhibits multiple protein family members, while
being potent against the target of interest, can be limited in its
utility as a human therapeutic due to unintended side effects and
toxicities introduced due to the consequences of inhibition of
these `off targets.`
[0006] Protein kinases constitute an important therapeutic protein
family. There are approximately 518 human protein kinases. While
inhibition of a desired kinase `on target` is desirable for a human
therapeutic, it is also desirable in many cases to provide a
selective kinase inhibitor which does not substantially inhibit
other kinase `off targets` from within this protein family.
Monoclonal antibodies are one approach to providing specific
inhibitors to a specific kinase without inhibiting `off targets.`
Achieving this level of selectivity with small molecule inhibitors,
however, is not as easily achievable nor as straightforward.
Accordingly, there is a need for kinase inhibitors that are
selective for a particular protein kinase. It is theorized that an
unexpected increase in potency for c-MET kinase inhibition or an
unexpected increase in selective c-MET inhibition relative to other
kinases is observed for one or more of the embodiments disclosed
herein.
SUMMARY
[0007] Compounds described herein find utility in the treatment of
mammalian cancers and especially human cancers including, but not
limited to, solid tumors, gastric cancers, melanomas,
glioblastomas, ovarian cancer, pancreatic cancer, prostate cancer,
lung cancers, non small cell lung cancer, breast cancers, kidney
cancers, cervical carcinomas, metastasis of primary tumor sites,
colonic cancers, myeloproliferative diseases, diseases wherein the
etiology or progression is dependent on c-MET kinase activity, or
on the activity of oncogenic forms, aberrant fusion protein forms,
and mutant forms of c-MET kinase.
[0008] Specifically, compounds of Formula I are disclosed which
find utility in the treatment of diseases as described above.
##STR00002##
[0009] In Formula I, X and F are regiochemically oriented with
respect to each other in a mutual para-orientation; X is halogen or
C1-C6 alkyl; and R3 is a non-hydrogen moiety regiochemically
oriented ortho- to the B ring nitrogen. Compounds described herein
exhibit unexpected potency for c-MET kinase inhibition and/or an
unexpected increase in selective c-MET kinase inhibition relative
to other kinases, particularly in comparison to other purported MET
kinase inhibitors.
[0010] In one aspect, compounds of the Formula I are described:
##STR00003##
[0011] and pharmaceutically acceptable salts, hydrates, solvates,
enantiomers, stereoisomers, and tautomers thereof;
[0012] wherein
[0013] X is halogen;
[0014] Z1 and Z2 are independently and individually CR2 or N;
[0015] Z3 is CH or N;
[0016] with the proviso that ring 13 is not a tetrazine;
[0017] each R1 is independently and individually halogen, H, C1-C6
alkyl, branched C3-C7 alkyl, C3-C7 cycloalkyl, or --CN;
[0018] each R2 is individually and independently H, halogen, C1-C6
alkyl, or cyano;
[0019] R3 is --C(O)R4, --C(O)--C6-C10-aryl,
--C(O)--C4-C6-heterocyclyl, or --C(O)--C5-C6-heteroaryl, wherein
[0020] aryl is phenyl, naphthyl, tetrahydronaphthyl, indenyl or
indanyl; and [0021] with the proviso that when R3 is
--C(O)--C4-C6-heterocyclyl, the heterocyclyl does not have a N
bonding hand to --C(O);
[0022] R4 is C1-C7 alkyl, C3-C8 cycloalkyl, --(CH.sub.2).sub.p--CN,
--(CH.sub.2).sub.p--OR6, --(CH.sub.2).sub.p--NR6(R7),
--(CH.sub.2).sub.p--SO.sub.2--C1-C6-alkyl,
--(CH.sub.2).sub.p--C6-C10-aryl,
--(CH.sub.2).sub.p--C5-C6-heteroaryl, or
--(CH.sub.2).sub.p--C4-C6-heterocyclyl, wherein [0023] each alkyl
or alkylene is optionally substituted with one or two C1-C6 alkyl;
and [0024] aryl is phenyl, naphthyl, tetrahydronaphthyl, indenyl or
indanyl;
[0025] each R6 and R7 is individually and independently H, C1-C6
alkyl, or C3-C8 branched alkyl;
[0026] each cycloalkyl, aryl, heteroaryl and heterocyclyl is
independently substituted with --(R25).sub.m;
[0027] each R25 is individually and independently C1-C6 alkyl,
branched C3-C8 alkyl, halogen, --(CH.sub.2).sub.m--CN,
--(CH.sub.2).sub.m--OR6, --(CH.sub.2).sub.m--NR6(R7),
--(CH.sub.2).sub.m--SO.sub.2--C1-C6-alkyl,
--(CH.sub.2).sub.m--C(O)NR6(R7),
--(CH.sub.2).sub.m--C(O)--C4-C6-heterocyclyl, or
--(CH.sub.2).sub.m--C4-C6-heterocyclyl, wherein each alkyl or
alkylene is optionally substituted with one or two C1-C6 alkyl;
[0028] each m is individually and independently 0, 1, 2, or 3;
and
[0029] p is 1, 2, or 3.
[0030] In some embodiments of the compound of Formula I, Z1 and Z2
are CR2, and Z3 is CH.
[0031] In certain embodiments, the compound is a compound of
Formula Ic,
##STR00004##
[0032] or a pharmaceutically acceptable salt, hydrate, solvate,
enantiomer, stereoisomer, or tautomer thereof, and
[0033] wherein n is 0, 1, or 2.
[0034] In certain embodiments of the compound of Formula Ic, R3 is
--C(O)R4.
[0035] In other embodiments of the compound of Formula Ic, R3 is
--C(O)R4 and R4 is C1-C7 alkyl, C3-C8 cycloalkyl,
--(CH.sub.2).sub.p--CN, --(CH.sub.2).sub.p--OR6,
--(CH.sub.2).sub.p--NR6(R7),
--(CH.sub.2).sub.p--SO.sub.2--C1-C6-alkyl, or
--(CH.sub.2).sub.p--C4-C6-heterocyclyl, and wherein each alkyl or
alkylene is optionally substituted with one or two C1-C6 alkyl.
[0036] In some embodiments of the compound of Formula Ic, R3 is
--C(O)R4 and R4 is C1-C7 alkyl or C3-C8 cycloalkyl, and wherein
each alkyl or alkylene is optionally substituted with one or two
C1-C6 alkyl.
[0037] In some embodiments of the compound of Formula Ic, R3 is
--C(O)--C6-C10-aryl, --C(O)--C4-C6-heterocyclyl, or
--C(O)--C5-C6-heteroaryl.
[0038] In some embodiments of the compound of Formula I, Z1 and Z2
are CR2, and Z3 is N.
[0039] In certain embodiments, the compound of Formula I is a
compound of Formula If
##STR00005##
[0040] or a pharmaceutically acceptable salt, hydrate, solvate,
enantiomer, stereoisomer, or tautomer thereof, and
[0041] wherein
[0042] n is 0, 1, or 2.
[0043] In certain embodiments of the compound of Formula If, R3 is
--C(O)R4.
[0044] In other embodiments of the compound of Formula If, R3 is
--C(O)--C6-C10-aryl, --C(O)--C4-C6-heterocyclyl, or
--C(O)--C5-C6-heteroaryl.
[0045] In some embodiments of the compound of Formula I, Z1 is CR2,
Z2 is N, and Z3 is CH.
[0046] In certain embodiments, the compound of Formula I is a
compound of Formula Ij,
##STR00006##
[0047] or a pharmaceutically acceptable salt, hydrate, solvate,
enantiomer, stereoisomer, or tautomer thereof.
[0048] In certain embodiments of the compound of Formula Ij, R3 is
--C(O)R4.
[0049] In other embodiments of the compound of Formula Ij, R3 is
--C(O)--C6-C10-aryl, --C(O)--C4-C6-heterocyclyl, or
--C(O)--C5-C6-heteroaryl.
[0050] In some embodiments of the compound of Formula I, Z1 is CR2,
and Z2 and Z3 are N.
[0051] In certain embodiments, the compound of Formula I is a
compound of Formula Im,
##STR00007##
or a pharmaceutically acceptable salt, hydrate, solvate,
enantiomer, stereoisomer, or tautomer thereof.
[0052] In some embodiments of the compound of Formula Im, R3 is
--C(O)R4.
[0053] In other embodiments of the compound of Formula Im, R3 is
--C(O)--C6-C10-aryl, --C(O)--C4-C6-heterocyclyl, or
--C(O)--C5-C6-heteroaryl.
[0054] In one embodiment, the present invention is directed to a
compound selected from the group consisting of
N-(4-(2-acetamidopyridin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluorophenyl)-
cyclopropane-1,1-dicarboxamide,
N-(4-(2-acetamidopyridin-4-yloxy)-5-chloro-2-fluorophenyl)-N'-(4-fluoroph-
enyl)cyclopropane-1,1-dicarboxamide,
N-(4-(2-acetamidopyridin-4-yloxy)-2,5-difluorophenyl)-N'-phenylcyclopropa-
ne-1,1-dicarboxamide,
N-(4-(2-(2-(dimethylamino)acetamido)pyridin-4-yloxy)-2,5-difluorophenyl)--
N'-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
N-(4-(2-acetamidopyrimidin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluoropheny-
l)cyclopropane-1,1-dicarboxamide,
N-(4-(2-(cyclopropanecarboxamido)pyridin-4-yloxy)-2,5-difluorophenyl)-N-(-
4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
N-(2,5-difluoro-4-(2-propionamidopyridin-4-yloxy)phenyl)-N'-(4-fluorophen-
yl)cyclopropane-1,1-dicarboxamide,
N-(2,5-difluoro-4-((2-(2-methoxyacetamido)pyridin-4-yl)oxy)phenyl)-N'-(4--
fluorophenyl)cyclopropane-1,1-dicarboxamide,
N-(2,5-difluoro-4-(2-isobutyramidopyridin-4-yloxy)phenyl)-N'-(4-fluorophe-
nyl)cyclopropane-1,1-dicarboxamide,
N-(4-(2-(2-cyanoacetamido)pyridin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluo-
rophenyl)cyclopropane-1,1-dicarboxamide,
N-(4-(2-(azetidine-3-carboxamido)pyridin-4-yloxy)-2,5-difluorophenyl)-N'--
(4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
N-(4-(2-(cyclobutanecarboxamido)pyridin-4-yloxy)-2,5-difluorophenyl)-N'-(-
4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
(R)--N-(2,5-difluoro-4-((2-(2-methoxypropanamido)pyridin-4-yl)oxy)phenyl)-
-N'-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
R)--N-(2,5-difluoro-4-((2-(2-hydroxypropanamido)pyridin-4-yl)oxy)phenyl)--
N'-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
N-(2,5-difluoro-4-((2-pivalamidopyridin-4-yl)oxy)phenyl)-N'-(4-fluorophen-
yl)cyclopropane-1,1-dicarboxamide,
(S)--N-(2,5-difluoro-4-((2-(2-methoxypropanamido)pyridin-4-yl)oxy)phenyl)-
-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
(S)-1-((4-(2,5-difluoro-4-(1-((4-fluorophenyl)carbamoyl)cyclopropanecarbo-
xamido)phenoxy)pyridin-2-yl)amino)-1-oxopropan-2-yl acetate,
N-(2,5-difluoro-4-((2-(2-fluoro-2-methylpropanamido)pyridin-4-yl)oxy)phen-
yl)-N'-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide and
pharmaceutically acceptable salts, solvates, hydrates and tautomers
thereof.
[0055] In another embodiment, the present invention is directed to
a compound selected from the group consisting of
N-(4-(2-(cyclopropanecarboxamido)pyridin-4-yloxy)-2,5-difluorophenyl)-N'--
(4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
N-(2,5-difluoro-4-(2-propionamidopyridin-4-yloxy)phenyl)-N'-(4-fluorophen-
yl)cyclopropane-1,1-dicarboxamide,
N-(2,5-difluoro-4-(2-isobutyramidopyridin-4-yloxy)phenyl)-N'-(4-fluorophe-
nyl)cyclopropane-1,1-dicarboxamide,
N-(4-(2-acetamidopyridin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluorophenyl)-
cyclopropane-1,1-dicarboxamide.
[0056] In some embodiments, the present invention comprises a
method of treating mammalian disease wherein the disease etiology
or progression is at least partially mediated by a kinase activity,
wherein the kinase is a wildtype form, a mutant oncogenic form, an
aberrant fusion protein form or a polymorph, the method comprising
administering to a mammal in need thereof an effective amount of a
compound of any of claims 1-21.
[0057] In certain embodiments, the disease etiology or progression
is at least partially mediated by the kinase activity of c-MET,
mutant oncogenic forms, aberrant fusion proteins, or polymorphs
thereof.
[0058] In some embodiments, the invention is directed to a
pharmaceutical composition, comprising a compound of any of claims
1-21 and a pharmaceutically acceptable carrier.
[0059] In certain embodiments, the pharmaceutical composition
further comprises an additive selected from adjuvants, excipients,
diluents, or stabilizers.
[0060] In other embodiments the present invention is directed to a
method of treating cancer, gastrointestinal stromal tumors,
hyperproliferative diseases, metabolic diseases, neurodegenerative
diseases, or diseases characterized by angiogenesis, such as solid
tumors, melanomas, glioblastomas, ovarian cancer; pancreatic
cancer, prostate cancer, lung cancers, breast cancers, renal
cancers, hepatic cancers, cervical carcinomas, metastasis of
primary tumor sites, myeloproliferative diseases, chronic
myelogenous leukemia, leukemias, papillary thyroid carcinoma,
non-small cell lung cancer, mesothelioma, hypereosinophilic
syndrome, colonic cancers, ocular diseases characterized by
hyperproliferation leading to blindness including retinopathies,
diabetic retinopathy, age-related macular degeneration,
hypereosinophilic syndrome, rheumatoid arthritis, asthma, chronic
obstructive pulmonary, mastocytosis, or mast cell leukemia, the
method comprising administering to a patient in need thereof an
effective amount of a compound of any of claims 1-21.
[0061] In some embodiments, the compound is administered orally,
parenterally, by inhalation, or subcutaneously.
[0062] The details of the invention are set forth in the
accompanying description below. Although methods and materials
similar or equivalent to those described herein can be used in the
practice or testing of the present invention, illustrative methods
and materials are now described. Other features, objects, and
advantages of the invention will be apparent from the description
and from the claims. In the specification and the appended claims,
the singular forms also include the plural unless the context
clearly dictates otherwise. Unless defined otherwise, all technical
and scientific terms used herein have the same meaning as commonly
understood by one of ordinary skill in the art to which this
invention belongs.
DETAILED DESCRIPTION
[0063] Throughout this disclosure, various patents, patent
applications and publications are referenced. The disclosures of
these patents, patent applications and publications in their
entireties are incorporated into this disclosure by reference in
order to more fully describe the state of the art as known to those
skilled therein as of the date of this disclosure. This disclosure
will govern in the instance that there is any inconsistency between
the patents, patent applications and publications and this
disclosure.
[0064] For convenience, certain terms employed in the
specification, examples and claims are collected here. Unless
defined otherwise, all technical and scientific terms used in this
disclosure have the same meanings as commonly understood by one of
ordinary skill in the art to which this disclosure belongs. The
initial definition provided for a group or term provided in this
disclosure applies to that group or term throughout the present
disclosure individually or as part of another group, unless
otherwise indicated.
[0065] The compounds of this disclosure include any and all
possible isomers, stereoisomers, enantiomers, diastereomers,
tautomers, pharmaceutically acceptable salts, and solvates thereof,
as well as crystalline polymorphic forms of the disclosed compounds
and any and all possible isomers, stereoisomers, enantiomers,
diastereomers, tautomers, pharmaceutically acceptable salts, and
solvates thereof. Thus, the terms "compound" and "compounds" as
used in this disclosure refer to the compounds of this disclosure
and any and all possible isomers, stereoisomers, enantiomers,
diastereomers, tautomers, pharmaceutically acceptable salts, and
solvates, and crystalline polymorphs thereof.
DEFINITIONS
[0066] The term "alkyl" as used herein refers to a straight chain
alkyl, wherein alkyl chain length is indicated by a range of
numbers. In exemplary embodiments, "alkyl" refers to an alkyl chain
as defined above containing 1, 2, 3, 4, 5, or 6 carbons (i.e.,
C1-C6 alkyl). Examples of an alkyl group include, but are not
limited to, methyl, ethyl, propyl, butyl, pentyl, and hexyl.
[0067] The term "branched alkyl" as used herein refers to an alkyl
chain wherein a branching point in the chain exists, and the total
number of carbons in the chain is indicated by a range of numbers.
In exemplary embodiments, "branched alkyl" refers to an alkyl chain
as defined above containing from 3, 4, 5, 6, 7, or 8 carbons (i.e.,
branched C3-C8 alkyl). Examples of a branched alkyl group include,
but are not limited to, iso-propyl, iso-butyl, secondary-butyl, and
tertiary-butyl.
[0068] The term "alkoxy" as used herein refers to --O-- (alkyl),
wherein "alkyl" is as defined above.
[0069] The term "branched alkoxy" as used herein refers to --O--
(branched alkyl), wherein "branched alkyl" is as defined above.
[0070] The term "alkylene" as used herein refers to an alkyl moiety
interposed between two other atoms. In exemplary embodiments,
"alkylene" refers to an alkyl moiety as defined above containing 1,
2, or 3 carbons. Examples of an alkylene group include, but are not
limited to --CH.sub.2--, --CH.sub.2CH.sub.2--, and
--CH.sub.2CH.sub.2CH.sub.2--. In exemplary embodiments, alkylene
groups are branched.
[0071] The term "alkynyl" as used herein refers to a carbon chain
containing one carbon-carbon triple bond. In exemplary embodiments,
"alkynyl" refers to a carbon chain as described above containing 2
or 3 carbons (i.e., C2-C3 alkynyl). Examples of an alkynyl group
include, but are not limited to, ethyne and propyne.
[0072] The term "aryl" as used herein refers to a cyclic
hydrocarbon, where the ring is characterized by delocalized .pi.
electrons (aromaticity) shared among the ring members, and wherein
the number of ring atoms is indicated by a range of numbers. In
exemplary embodiments, "aryl" refers to a cyclic hydrocarbon as
described above containing 6, 7, 8, 9, or ring atoms (i.e., C6-C10
aryl). Examples of an aryl group include, but are not limited to,
benzene, naphthalene, tetralin, indene, and indane.
[0073] The term "cycloalkyl" as used herein refers to a monocyclic
saturated carbon ring, wherein the number of ring atoms is
indicated by a range of numbers. In exemplary embodiments,
"cycloalkyl" refers to a carbon ring as defined above containing 3,
4, 5, 6, 7, or 8 ring atoms (i.e., C3-C8 cycloalkyl). Examples of a
cycloalkyl group include, but are not limited to, cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and
cyclooctyl.
[0074] The term "halogen" as used herein refers to fluorine,
chlorine, bromine, and iodine.
[0075] The term "heterocycle" or "heterocyclyl" as used herein
refers to a cyclic hydrocarbon, wherein at least one of the ring
atoms is an O, N, or S, wherein the number of ring atoms is
indicated by a range of numbers. Heterocyclyl moieties as defined
herein have C or N bonding hands. For example, in some embodiments,
a ring N atom from the heterocyclyl is the bonding atom to --C(O)
to form an amide, carbamate, or urea. In exemplary embodiments,
"heterocyclyl" refers to a cyclic hydrocarbon as described above
containing 4, 5, or 6 ring atoms (i.e., C4-C6 heterocyclyl).
Examples of a heterocycle group include, but are not limited to,
aziridine, oxirane, thiirane, azetidine, oxetane, thietane,
pyrrolidine, tetrahydrofuran, pyran, thiopyran, thiomorpholine,
thiomorpholine S-oxide, thiomorpholine'S-dioxide, oxazoline,
tetrahydrothiophene, piperidine, tetrahydropyran, thiane,
imidazolidine, oxazolidine, thiazolidine, dioxolane, dithiolane,
piperazine, oxazine, dithiane, and dioxane.
[0076] The term "heteroaryl" as used herein refers to a cyclic
hydrocarbon, where at least one of the ring atoms is an O, N, or S,
the ring is characterized by delocalized it electrons (aromaticity)
shared among the ring members, and wherein the number of ring atoms
is indicated by a range of numbers. Heteroaryl moieties as defined
herein have C or N bonding hands. For example, in some embodiments,
a ring N atom from the heteroaryl is the bonding atom to --C(O) to
form an amide, carbamate, or urea. In exemplary embodiments,
"heteroaryl" refers to a cyclic hydrocarbon as described above
containing 5 or 6 ring atoms (i.e., C5-C6 heteroaryl). Examples of
a heteroaryl group include, but are not limited to, pyrrole, furan,
thiene, oxazole, thiazole, isoxazole, isothiazole, imidazole,
pyrazole, oxadiazole, thiadiazole, triazole, tetrazole, pyridine,
pyrimidine, pyrazine, pyridazine, and triazine.
[0077] The term "substituted" in connection with a moiety as used
herein refers to a further substituent which is attached to the
moiety at any acceptable location on the moiety. Unless otherwise
indicated, moieties can bond through a carbon, nitrogen, oxygen,
sulfur, or any other acceptable atom.
[0078] The term "salts" as used herein embraces pharmaceutically
acceptable salts commonly used to form alkali metal salts of free
acids and to form addition salts of free bases. The nature of the
salt is not critical, provided that it is pharmaceutically
acceptable. Suitable pharmaceutically acceptable acid addition
salts may be prepared from an inorganic acid or from an organic
acid. Exemplary pharmaceutical salts are disclosed in Stahl, P. H.,
Wermuth, C. C., Eds. Handbook of Pharmaceutical Salts: Properties,
Selection and Use; Verlag Helvetica Chimica Acta/Wiley-VCH: Zurich,
2002, the contents of which are hereby incorporated by reference in
their entirety. Specific non-limiting examples of inorganic acids
are hydrochloric, hydrobromic, hydroiodic, nitric, carbonic,
sulfuric and phosphoric acid. Appropriate organic acids include,
without limitation, aliphatic, cycloaliphatic, aromatic,
arylaliphatic, and heterocyclyl containing carboxylic acids and
sulfonic acids, for example formic, acetic, propionic, succinic,
glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic,
glucuronic, maleic, fumaric, pyruvic, aspartic, glutamic, benzoic,
anthranilic, mesylic, stearic, salicylic, p-hydroxybenzoic,
phenylacetic, mandelic, embonic (pamoic), methanesulfonic,
ethanesulfonic, benzenesulionic, pantothenic, toluenesulfonic,
2-hydroxyethanesulfonic, sulfanilic, cyclohexylaminosulfonic,
algenic, 3-hydroxybutyric, galactaric or galacturonic acid.
Suitable pharmaceutically acceptable salts of free acid-containing
compounds disclosed herein include, without limitation, metallic
salts and organic salts. Exemplary metallic salts include, but are
not limited to, appropriate alkali metal (group Ia) salts, alkaline
earth metal (group IIa) salts, and other physiological acceptable
metals. Such salts can be made from aluminum, calcium, lithium,
magnesium, potassium, sodium and zinc. Exemplary organic salts can
be made from primary amines, secondary amines, tertiary amines and
quaternary ammonium salts, for example, tromethamine, diethylamine,
tetra-N-methylammonium, N,N'-dibenzylethylenediamine,
chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine
(N-methylglucamine) and procaine.
[0079] The term "salts" as used herein embraces pharmaceutically
acceptable salts commonly used to form alkali metal salts of free
acids and to form addition salts of free bases. The nature of the
salt is not critical, provided that it is pharmaceutically
acceptable. Suitable pharmaceutically acceptable acid addition
salts may be prepared from an inorganic acid or from an organic
acid. Examples of such inorganic acids are hydrochloric,
hydrobromic, hydroiodic, nitric, carbonic, sulfuric and phosphoric
acid. Appropriate organic acids may be selected from aliphatic,
cycloaliphatic, aromatic, arylaliphatic, and heterocyclyl
containing carboxylic acids and sulfonic acids, for example formic,
acetic, propionic, succinic, glycolic, gluconic, lactic, malic,
tartaric, citric, ascorbic, glucuronic, maleic, fumaric, pyruvic,
aspartic, glutamic, benzoic, anthranilic, mesylic, stearic,
salicylic, p-hydroxybenzoic, phenylacetic, mandelic, embonic
(pamoic), methanesulfonic, ethanesulfonic, benzenesulfonic,
pantothenic, toluenesulfonic, 2-hydroxyethanesulfonic, sulfanilic,
cyclohexylaminosulfonic, algenic, 3-hydroxybutyric, galactaric and
galacturonic acid. Suitable pharmaceutically acceptable salts of
free acid-containing compounds disclosed herein include metallic
salts and organic salts. Exemplary metallic salts include, but are
not limited to, appropriate alkali metal (group Ia) salts, alkaline
earth metal (group IIa) salts, and other physiological acceptable
metals. Such salts can be made from aluminum, calcium, lithium,
magnesium, potassium, sodium and zinc. Exemplary organic salts can
be made from primary amines, secondary amines, tertiary amines and
quaternary ammonium salts, for example, tromethamine, diethylamine,
tetra-N-methylammonium, N,N'-dibenzylethylenediamine,
chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine
(N-methylglucamine) and procaine.
[0080] The terms "administer," "administering" or "administration"
as used herein refer to either directly administering a compound or
pharmaceutically acceptable salt of the compound or a composition
to a subject.
[0081] The term "carrier" as used herein encompasses carriers,
excipients, and diluents, meaning a material, composition or
vehicle, such as a liquid or solid filler, diluent, excipient,
solvent or encapsulating material involved in carrying or
transporting a pharmaceutical agent from one organ, or portion of
the body, to another organ or portion of the body.
[0082] The term "disorder" is used in this disclosure to mean, and
is used interchangeably with, the terms disease, condition, or
illness, unless otherwise indicated.
[0083] The terms "effective amount" and "therapeutically effective
amount" are used interchangeably in this disclosure and refer to an
amount of a compound that, when administered to a subject, is
capable of reducing a symptom of a disorder in a subject. The
actual amount which comprises the "effective amount" or
"therapeutically effective amount" will vary depending on a number
of conditions including, but not limited to, the particular
disorder being treated, the severity of the disorder, the size and
health of the patient, and the route of administration. A skilled
medical practitioner can readily determine the appropriate amount
using methods known in the medical arts.
[0084] The terms "isolated" and "purified" as used herein refer to
a component separated from other components of a reaction mixture
or a natural source. In certain embodiments, the isolate contains
at least about 50%, at least about 55%, at least about 60%, at
least about 65%, at least about 70%, at least about 75%, at least
about 80%, at least about 85%, at least about 90%, at least about
95%, or at least about 98% of the compound or pharmaceutically
acceptable salt of the compound by weight of the isolate.
[0085] The phrase "pharmaceutically acceptable" as used herein
refers to those compounds, materials, compositions, and/or 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, allergic response,
or other problem or complication, commensurate with a reasonable
benefit/risk ratio:
[0086] As used in this disclosure, the term "subject" includes,
without limitation, a human or an animal. Exemplary animals
include, but are not limited to, mammals such as mouse, rat, guinea
pig, dog, cat, horse, cow, pig, monkey, chimpanzee, baboon, or
rhesus monkey.
[0087] The term "treating" as used herein with regard to a subject,
refers to improving at least one symptom of the subject's disorder.
Treating can be curing, improving, or at least partially
ameliorating the disorder.
[0088] The term "hydrate" as used herein refers to a compound
disclosed herein which is associated with water in the molecular
form, i.e., in which the H--OH bond is not split, and may be
represented, for example, by the formula RH.sub.2O, where R is a
compound disclosed herein. A given compound may form more than one
hydrate including, for example, monohydrates (RH.sub.2O),
dihydrates (R2H.sub.2O), trihydrates (R3H.sub.2O), and the
like.
[0089] The term "solvate" as used herein refers to a compound
disclosed herein which is associated with solvent in the molecular
form, i.e., in which the solvent is coordinatively hound, and may
be represented, for example, by the formula R(solvent), where R is
a compound disclosed herein. A given compound may form more than
one solvate including, for example, monosolvates (R(solvent)) or
polysolvates (Rn(solvent)), wherein n is an integer greater than 1)
including, for example, disolvates (R2(solvent)), trisolvates
(R3(solvent)), and the like, or hemisolvates, such as, for example,
Rn/2(solvent), Rn/3(solvent), Rn/4(solvent) and the like, wherein n
is an integer. Solvents herein include mixed solvents, for example,
methanol/water, and as such, the solvates may incorporate one or
more solvents within the solvate.
[0090] The term "acid hydrate" as used herein refers to a complex
that may be formed through association of a compound having one or
more base moieties with at least one compound having one or more
acid moieties or through association of a compound having one or
more acid moieties with at least one compound having one or more
base moieties, said complex being further associated with water
molecules so as to form a hydrate, wherein said hydrate is as
previously defined and R represents the complex herein described
above.
[0091] Structural, chemical and stereochemical definitions are
broadly taken from IUPAC recommendations, and more specifically
from Glossary of Terms used in Physical Organic Chemistry (IUPAC
Recommendations 1994) as summarized by Muller, P. Pure Appl. Chem.
1994, 66, pp. 1077-1184 and Basic Terminology of Stereochemistry
(IUPAC Recommendations 1996) as summarized by Moss, G. P. Pure
Appl. Chem. 1996, 68, pp. 2193-2222.
[0092] Atropisomers are defined as a subclass of conformers which
can be isolated as separate chemical species and which arise from
restricted rotation about a single bond.
[0093] Regioisomers or structural isomers are defined as isomers
involving the same atoms in different arrangements.
[0094] Enantiomers are defined as one of a pair of molecular
entities which are mirror images of each other and
non-superimposable.
[0095] Diastereomers or diastereoisomers are defined as
stereoisomers other than enantiomers. Diastereomers or
diastereoisomers are stereoisomers not related as mirror images.
Diastereoisomers are characterized by differences in physical
properties, and by some differences in chemical behavior towards
achiral as well as chiral reagents.
[0096] The term "tautomer" as used herein refers to compounds
produced by the phenomenon wherein a proton of one atom of a
molecule shifts to another atom. See March, Advanced Organic
Chemistry: Reactions, Mechanisms and Structures, 4th Ed., John
Wiley & Sons, pp. 69-74 (1992). Tautomerism is defined as
isomerism of the general form
G-X--Y.dbd.ZX.dbd.Y--Z-G
where the isomers (called tautomers) are readily interconvertible;
the atoms connecting the groups X, Y and Z are typically any of C,
H, O, or S, and G is a group which becomes an electrofuge or
nucleofuge during isomerization. The most common case, when the
electrofuge is H.sup.+, is also known as "prototropy." Tautomers
are defined as isomers that arise from tautomerism, independent of
whether the isomers are isolable.
[0097] ChemDraw version 8.0 or 10, (CambridgeSoft Corporation,
Cambridge, Mass.) was used to name structures.
[0098] The following abbreviations are used in this disclosure and
have the following definitions: ADP is adenosine diphosphate, ATP
is adenosine triphosphate, dba is dibenzylideneacetone, DIEA is
N,N-diisopropylethylamine, DMA is N,N-dimethylacetamide, DMF is
N,N-dimethylformamide, DMSO is dimethylsulfoxide, DTT is
dithiothreitol, EGTA is ethylene glycol tetraacetic acid, ESI is
electrospray ionization, GST is glutathione S-transferase, "h" is
hour or hours, HATU is
2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate, HEPES is
4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; HPLC is high
pressure. (or performance) liquid chromatography, IC.sub.50 is half
maximal inhibitory concentration, MS is mass spectrometry, min is
minutes, NADH is nicotinamide adenine dinucleotide, NMR is nuclear
magnetic resonance, PBS is phosphate buffered saline, RT is room
temperature, THF is tetrahydrofuran, Tris is
tris(hydroxymethyl)aminomethane, and xantphos is
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene.
Compounds
[0099] In one aspect, compounds of the Formula I are described:
##STR00008##
[0100] and pharmaceutically acceptable salts, hydrates, solvates,
enantiomers, stereoisomers, and tautomers thereof;
[0101] wherein
[0102] X, B, Z1, Z2, Z3, R1, R2, R3, R4, R6, R7, R25, m, n and p
are as defined above for Formula I; and
[0103] each heterocyclyl and heteroaryl individually and
independently has a C or N bonding hand.
[0104] In some embodiments, a ring N atom from the heterocyclyl is
the bonding atom to --C(O) to form an amide, carbamate, or urea. In
other embodiments, a ring N atom from the heteroaryl is the bonding
atom to --C(O) to form an amide, carbamate, or urea.
[0105] In some embodiments, compounds of the Formula I are
compounds of the Formula Ib:
##STR00009##
[0106] and pharmaceutically acceptable salts, hydrates, solvates,
enantiomers, stereoisomers, and tautomers thereof;
[0107] wherein
[0108] X, R1, R2, R3, R4, R6, R7, R25, m, n and p are as defined
above for Formula I; and
[0109] n is 0, 1, or 2;
[0110] In some embodiments, compounds of the Formula I are
compounds of the Formula Ic:
##STR00010##
[0111] and pharmaceutically acceptable salts, hydrates, solvates,
enantiomers, stereoisomers, and tautomers thereof;
[0112] wherein
[0113] R2, R3, R4, R6, R7, R25, m, n, and p are as defined above
for Formula Ib.
[0114] In some embodiments, compounds of the Formula I are
compounds of the Formula Ie:
##STR00011##
[0115] and pharmaceutically acceptable salts, hydrates, solvates,
enantiomers, stereoisomers, and tautomers thereof;
[0116] wherein
[0117] X, R1, R2, R3, R4, R6, R7, R25, m, n, and p are as defined
above for Formula Ib.
[0118] In some embodiments, compounds of the Formula I are
compounds of the Formula If:
##STR00012##
[0119] and pharmaceutically acceptable salts, hydrates, solvates,
enantiomers, stereoisomers, and tautomers thereof;
[0120] wherein
[0121] R2, R3, R4, R6, R7, R25, m, n, and p are as defined above
for Formula Ib.
[0122] In some embodiments, compounds of the Formula I are
compounds of the Formula Ih:
##STR00013##
[0123] and pharmaceutically acceptable salts, hydrates, solvates,
enantiomers, stereoisomers, and tautomers thereof;
[0124] wherein
[0125] X, R1, R2, R3, R4, R6, R7, R25, m, n and p are as defined
above for Formula I.
[0126] In some embodiments, compounds of the Formula I are
compounds of the Formula Ij:
##STR00014##
[0127] and pharmaceutically acceptable salts, hydrates, solvates,
enantiomers, stereoisomers, and tautomers thereof;
[0128] wherein
[0129] R2, R3, R4, R6, R7, R25, m, n and p are as defined above for
Formula I.
[0130] In some embodiments, compounds of the Formula I are
compounds of the Formula Il:
##STR00015##
[0131] and pharmaceutically acceptable salts, hydrates, solvates,
enantiomers, stereoisomers, and tautomers thereof;
[0132] wherein
[0133] X, R1, R2, R3, R4, R6, R7, R25, m, n and p are as defined
above for Formula I.
[0134] In some embodiments, compounds of the Formula I are
compounds of the Formula Im:
##STR00016##
[0135] and pharmaceutically acceptable salts, hydrates, solvates,
enantiomers, stereoisomers, and tautomers thereof;
[0136] wherein
[0137] R2, R3, R4, R6, R7, R25, m, n and p are as defined above for
Formula I.
[0138] The following embodiments are descriptive of Formula I,
Formula Ib, Formula Ie, Formula Ib, and Formula Il.
[0139] In some embodiments, X is halogen. In other embodiments, X
is F or Cl. In further embodiments, X is F.
[0140] In some embodiments, each R1 is individually and
independently halogen. In other embodiments, each R1 is
individually and independently F or Cl. In further embodiments,
each R1 is F.
[0141] In some embodiments, m is 1 and R1 is halogen. In other
embodiments, m is 1 and R1 is F or Cl. In further embodiments, m is
1 and R1 is F.
[0142] In some embodiments, X and each R1 is individually and
independently halogen. In other embodiments, X and each R1 is
individually and independently F or Cl. In further embodiments, X
and each R1 is F.
[0143] In some embodiments, m is 1 and X and each R1 is
individually and independently halogen. In other embodiments, m is
1 and X and each R1 is individually and independently F or Cl. In
further embodiments m is 1 and X and each R1 is F.
[0144] The following embodiments are descriptive of Formula I,
Formula Ib, Formula Ic, Formula Ie, Formula If, Formula Ih, Formula
Ij, Formula Il, and Formula Im.
[0145] In some embodiments, R3 is --C(O)R4 and R4 is C1-C7 alkyl,
C3-C8 cycloalkyl, --(CH.sub.2).sub.p--CN, --(CH.sub.2).sub.p--OR6,
--(CH.sub.2).sub.p--NR6(R7), or
--(CH.sub.2).sub.p--C4-C6-heterocyclyl, wherein each alkyl or
alkylene is optionally substituted with one or two C1-C6 alkyl. In
further embodiments, one alkyl or alkylene is substituted by one
C1-C6 alkyl. In still further embodiments, one alkyl or alkylene is
substituted by one C1-alkyl.
[0146] In some embodiments, R3 is --C(O)--C6-C10-aryl,
--C(O)--C4-C6-heterocyclyl, or --C(O)--C5-C6-heteroaryl.
[0147] In illustrative embodiments, compounds disclosed herein are
as set forth below.
##STR00017##
UTILITY
[0148] Compounds described herein rind utility in the treatment of
mammalian cancers and especially human cancers including, but not
limited to, solid tumors, gastric cancers, melanomas,
glioblastomas, ovarian cancer, pancreatic cancer, prostate cancer,
lung cancers, non small cell lung cancer, breast cancers, kidney
cancers, cervical carcinomas, metastasis of primary tumor sites,
colonic cancers, myeloproliferative diseases, diseases wherein the
etiology or progression is dependent on c-MET kinase activity, or
on the activity of oncogenic forms-, aberrant fusion protein forms,
and mutant forms of c-MET kinase.
Administration of Compounds
[0149] In some embodiments, the compound is administered by a
method selected from the group consisting of oral, parenteral,
inhalation, and subcutaneous.
Treatment Methods
[0150] The disclosed methods also include treating individuals
suffering from a condition selected from the group consisting of
cancer, hyperproliferative diseases, metabolic diseases,
neurodegenerative diseases or diseases characterized by
angiogenesis. These methods comprise administering to such
individuals compounds disclosed herein, and especially those of
section 1, said diseases including, but not limited to, solid
tumors, malignant melanomas, glioblastomas, ovarian cancer,
pancreatic cancer, prostate cancer, lung cancers, breast cancers,
kidney cancers, hepatic cancers, cervical carcinomas, metastasis of
primary tumor sites, myeloproliferative diseases, chronic
myelogenous leukemia, leukemias, papillary thyroid carcinoma,
non-small cell lung cancer, mesothelioma, hypereosinophilic
syndrome, gastrointestinal stromal tumors, colonic cancers, ocular
diseases characterized by hyperproliferation leading to blindness
including various retinopathies, diabetic retinopathy and
age-related macular degeneration and hypereosinophilic syndrome,
rheumatoid arthritis, asthma, chronic obstructive pulmonary
disorder, mastocytosis, mast cell leukemia, a disease caused by
c-MET kinase, oncogenic forms thereof, aberrant fusion proteins
thereof and polymorphs thereof. The administration method is not
critical, and may be from the group consisting of oral, parenteral,
inhalation, and subcutaneous.
Pharmaceutical Preparations
[0151] The compounds disclosed herein may form a part of a
pharmaceutical composition by combining one or more such compounds
with a pharmaceutically acceptable carrier. Additionally, the
compositions may include an additive selected from the group
consisting of adjuvants, excipients, diluents, and stabilizers.
Methods of Making
[0152] The compounds of the invention are available by the general
synthetic methods illustrated in the Schemes below and the
accompanying examples.
[0153] Compounds 1 of the invention are assembled in a step-wise
manner as illustrated in Scheme 1. Beginning with
cyclopropane-1,1-dicarboxylic-acid 2, standard peptide coupling
chemistry familiar to those skilled in the art is employed in the
formation of a new amide bond with amine 3 to yield intermediate 4.
Alternatively, it is recognized that in this case and in others to
follow, a carboxylic acid moiety, such as found in 2 is masked as
an ester or activated as an acid halide, anhydride, mixed
anhydride, or as an activated ester. In the case of activated acid
derivatives it should be understood that these compounds are
optionally isolated as discrete intermediates prior to their union
with amines-3 to form 4. Subsequent coupling of 4 with aniline 5,
either by peptide coupling conditions or via an activated acid
intermediate, yields compounds of the desired formula 1. Using
similar strategies, carboxylic acid 2 is also, in some embodiments,
first coupled with aniline 5 to yield intermediate 6, which is then
in turn coupled with 3 to yield desired compound 1.
##STR00018##
[0154] Non-limiting examples of the strategies described in Scheme
1 are illustrated below. Scheme 2 illustrates the preparation of
compound 10, an example of general formula 1 (wherein R1 is F, Z1,
Z2, and Z3 are CH and R3 is --C(O)CH.sub.3) by the general sequence
of 2.fwdarw.4.fwdarw.1 (Scheme 1). Thus, as indicated below, the
union of 1,1-cyclopropane his-carboxylic acid 2 with amine 7 (an
example of general amine 3) provides the amide/acid 8, an example
of general intermediate 4. Conditions for the transformation
include the in situ activation of bis-acid 2 by treatment with
thionyl chloride in the presence of a tertiary base, such as
triethylamine, followed by reaction with amine 7. Further reaction
of 8 with amine 9 (an example of general intermediate 5) in the
presence of a peptide coupling agent provides bis-amide 10.
Coupling agents for the later transformation include TBTU
(O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
tetrafluoroborate), PyBOP
(benzotriazol-1-yloxy)tripyrrolidinophosphonium
hexafluorophosphate), EDC
(1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride) and
BOP-Cl (bis(2-oxo-3-oxazolidinyl)phosphonic chloride).
##STR00019##
[0155] An example of an alternate route to 10, an example of
general formula 1, is shown in Scheme 3. In this case the
preparation begins with 11, in which one carboxylic acid moiety of
dicarboxylic acid 2 is protected as a methyl ester. Using the
conditions described above (Scheme 1) acid 11 and aniline 9 are
coupled to yield methyl ester 12. Saponification of ester 12 using
standard conditions (e.g., aqueous LiOH), followed by treatment
with thionyl chloride, yields the activated acid chloride
intermediate 13. Acid chloride 13 readily reacts with amine 7 in
the presence of a base such as triethylamine or Hunig's base to
yield example 10.
##STR00020##
[0156] Amines of the general formula 3 that are useful for the
invention are prepared by standard methods familiar to those
skilled in the art. Several non-limiting examples are shown in the
following schemes. A mixture of phenol 14 and benzamide 15, wherein
LG is a leaving group such as a halide or sulfonate, are coupled in
the presence of a base such as potassium tert-butoxide and a polar
aprotic solvent at elevated temperatures (e.g., 100.degree. C.) to
yield 16 (Scheme 4). Protection of the aniline NH.sub.2 of 16 with
the appropriate protecting group (PG) familiar to one skilled in
the art, such as a tert-butoxycarbonyl (BOC) group, followed by
subjection to Hofmann rearrangement conditions results in the
formation of 17. Acylation of 17 with R3-LG 18, followed by removal
of the protecting group yields amine 3. In one instance, the
reagent R3-LG (18) is a carboxylic acid (wherein LG is OH) that is
coupled with the amino moiety of 17 using standard peptide Coupling
agents, as described above. Alternately, reagent R3-LG (18) is an
activated carboxylic acid derivative, such as an acid halide
(wherein LG is halo) that undergoes reaction with amine 17 to
provide 3.
##STR00021##
[0157] A non-limiting example of this synthetic route is shown in
Scheme 5. Thus, the coupling of phenol 14 with 4-chloropicolinamide
19 (an example of intermediate 15, see Scheme 4) wherein Z1, Z2,
and Z3 are CH, and LG is Cl) is effected by heating in the presence
of a base to yield 20. Protection of the aniline moiety of 20 with
a BOC group using conditions familiar to one skilled in the art
affords 21. Amide 21 in turn undergoes a Hofmann rearrangement to
yield aminopyridine 22. Conditions for the Hofmann rearrangement
include bromine in aqueous KOH or addition of oxidants such as lead
tetraacetate or hypervalent iodine reagents such as
bis(trifluoroacetyl)iodobenzene in pyridine. Subsequent acylation
of 22 with acetyl chloride (an example of R3-LG wherein LG is
chloro) in a solution of pyridine yields 23. Removal of the BOC
protecting group in a solution of HCl provides amine 7, an example
of amine 3 wherein Z1, Z2, and Z3 are CH and R3 is
--C(O)CH.sub.3.
##STR00022##
[0158] Alternately, a modified version of the route illustrated in
Scheme 4 is shown in Scheme 6. The synthesis of 16, vide supra, is
followed by its union with carboxylic acid 6 to yield 24 using
either peptide coupling chemistry or an activated acid analog of 6.
Subjecting 24 to Hofmann rearrangement conditions yields 25, which
is then acylated with activated acid 18 to yield compounds of
formula 1.
##STR00023##
[0159] Amines of the general formula 3 are also accessed via 26
wherein Y is a typical leaving group in transition metal mediated
coupling reactions (for example, chloride, bromide, or triflate)
(Scheme 7). Treatment of 26 and amide 27 in an aprotic solvent, for
example 1,4-dioxane, with a catalytic amount of Pd(OAc).sub.2 or
Pd.sub.2(dba).sub.3 and xantphos in the presence of cesium
carbonate at elevated temperatures between 45.degree. C. and
110.degree. C. yields intermediate 3 (see Buchwald, et. al. Org.
Lett. (2000), 2(8): 1101). Similarly, intermediate 28 is assembled
from 26 and 6 using methods described in Scheme 3 and subsequently
reacted with 27 using catalytic palladium and xantphos (vide supra)
to yield compounds of formula 1.
##STR00024##
[0160] Amine 26 is synthesized in a variety of ways, including
those shown below in the following non-limiting examples. As
depicted, in Scheme 8, amino-phenol 14 and 29 (wherein LG is a
leaving group in a nucleophilic substitution reaction, such as a
halide or sulfonate) is coupled upon addition of a base such as
potassium tert-butoxide in a solution of DMA at elevated
temperatures of 80.degree. C. to 100.degree. C.
##STR00025##
[0161] General amine 26 is also accessed via the
1-fluoro-4-nitrobenzene intermediate 30 (Scheme 9). The coupling of
30 with 31 proceeds at temperatures ranging from 0.degree. C. to
80.degree. C. in the presence of a base, for example sodium
hydride. The resultant nitro intermediate 32 is then reduced using
a variety of methods familiar to one skilled in the art to afford
amine 26.
##STR00026##
[0162] A non-limiting example of Scheme 9 is illustrated below for
the synthesis of 36, a specific example of 26 wherein X is F, Y is
Cl, and Z1, Z2, and Z3 are CH (Scheme 10). Addition of
1,2,4-trifluoro-5-nitrobenzene (33) to a solution of
2-chloropyridin-4-ol (34) and sodium hydride in DMF at 0.degree. C.
yields the nitro intermediate 35. The nitro moiety of 35 is
subsequently reduced at RT in the presence of zinc dust and
ammonium chloride in solution of methanol and THF to yield amine
36.
##STR00027##
[0163] A non-limiting example of Scheme 7 is illustrated in Scheme
11, beginning with intermediate 36, prepared in Scheme 10. Thus, 36
readily reacts with acid chloride 13 (see. Scheme 3) in the
presence of triethylamine to yield chloro-pyridine 37.
Chloro-pyridine 37 is then converted to 38, a specific example of 1
wherein R1 is F, X is F, Z1, Z2, and Z3 are CH and R3 is
--C(O)CH.sub.3, upon treatment with acetamide (an example of
R3-NH.sub.2 27 where R3 is acetyl) and cesium carbonate in the
presence of a catalytic amount of palladium acetate and
xantphos.
##STR00028##
[0164] Using the synthetic procedures and methods described herein
and methods known to those skilled in the art, the following
compounds were made:
N-(4-(2-acetamidopyridin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluorop-
henyl)cyclopropane-1,1-dicarboxamide,
N-(4-(2-acetamidopyridin-4-yloxy)-5-chloro-2-fluorophenyl)-N'-(4-fluoroph-
enyl)cyclopropane-1,1-dicarboxamide,
N-(4-(2-acetamidopyridin-4-yloxy)-2,5-difluorophenyl)-N'-phenylcyclopropa-
ne-1,1-dicarboxamide,
N-(4-(2-(2-(dimethylamino)acetamido)pyridin-4-yloxy)-2,5-difluorophenyl)--
N'-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
N-(4-(2-acetamidopyrimidin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluoropheny-
l)cyclopropane-1,1-dicarboxamide,
N-(4-(2-(cyclopropanecarboxamido)pyridin-4-yloxy)-2,5-difluorophenyl)-N'--
(4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
N-(2,5-difluoro-4-(2-propionamidopyridin-4-yloxy)phenyl)-N'-(4-fluorophen-
yl)cyclopropane-1,1-dicarboxamide,
N-(2,5-difluoro-4-((2-(2-methoxyacetamido)pyridin-4-yl)oxy)phenyl)-N'-(4--
fluorophenyl)cyclopropane-1,1-dicarboxamide,
N-(2,5-difluoro-4-(2-isobutyramidopyridin-4-yloxy)phenyl)-N'-(4-fluorophe-
nyl)cyclopropane-1,1-dicarboxamide,
N-(4-(2-(2-cyanoacetamido)pyridin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluo-
rophenyl)cyclopropane-1,1-dicarboxamide,
N-(4-(2-(azetidine-3-carboxamido)pyridin-4-yloxy)-2,5-difluorophenyl)-N'--
(4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
N-(4-(2-(cyclobutanecarboxamido)pyridin-4-yloxy)-2,5-difluorophenyl)-N'-(-
4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
(R)--N-(2,5-difluoro-4-((2-(2-methoxypropanamido)pyridin-4-yl)oxy)phenyl)-
-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
R)--N-(2,5-difluoro-4-((2-(2-hydroxypropanamido)pyridin-4-yl)oxy)phenyl)--
N'-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide,
N-(2,5-difluoro-4-((2-pivalamidopyridin-4-yl)oxy)phenyl)-N'-(4-fluorophen-
yl)cyclopropane-1,1-dicarboxamide, (S)--N-(2,5-d
fluoro-4-((2-(2-methoxypropanamido)pyridin-4-yl)oxy)phenyl)-N-(4-fluoroph-
enyl)cyclopropane-1,1-dicarboxamide,
(S)-1-((4-(2,5-difluoro-4-(1-((4-fluorophenyl)carbamoyl)cyclopropanecarbo-
xamido)phenoxy)pyridin-2-yl)amino)-1-oxopropan-2-yl acetate, and
N-(2,5-difluoro-4-((2-(2-fluoro-2-methylpropanamido)pyridin-4-yl)oxy)phen-
yl)-N'-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide.
EXAMPLES
[0165] The disclosure is further illustrated by the following
examples, which are not to be construed as limiting this disclosure
in scope or spirit to the specific procedures herein described. It
is to be understood that the examples are provided to illustrate
certain embodiments and that no limitation to the scope of the
disclosure is intended thereby. It is to be further understood that
resort may be had to various other embodiments, modifications, and
equivalents thereof which may suggest themselves to those skilled
in the art without departing from the spirit of the present
disclosure and/or scope of the appended claims.
Example A1
[0166] Sodium hydride (60% by weight in mineral oil) (3.08 g, 77
mmol) was placed in a 500 mL round bottom flask flushed with argon.
DMF (140 mL) was added and the mixture was cooled in an ice bath.
2-Chloro-4-hydroxypyridine (7.68 g, 59.3 mmol) was then added
slowly over 45 minutes. After addition of the hydroxypyridine was
complete 2,4,5-trifluoronitrobenzene (10.5 g, 59.3 mmol) was added
as a solution in DMF (29 mL). The mixture was warmed to room
temperature and stirred for 18 hours. The reaction mixture was
concentrated under reduced pressure to remove the majority of DMF
in the mixture, and was then partitioned between ethyl acetate (300
mL) and 10% aqueous lithium chloride (150 mL). A precipitate formed
which was removed via suction filtration and then the layers were
separated. The organic layer was washed with additional 10% aqueous
lithium chloride (2.times.150 mL), saturated aqueous sodium
bicarbonate (150 mL) and brine (150 mL). The organic layer was
dried over magnesium sulfate and evaporated to yield a dark red
solid which was purified by silica gel chromatography (10 to 30%
ethyl acetate/hexane) to give
2-chloro-4-(2,5-difluoro-4-nitrophenoxy)pyridine (13.56 g, 80%
yield) as a yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6):
.delta. 8.45 (dd, 1H), 8.39 (d, 1H), 7.87 (dd, 1H), 7.39 (d, 1H),
7.24 (dd, 1H); MS (ESI) m/z: 287.0 (M+H.sup.+).
[0167] 2-Chloro-4-(2,5-difluoro-4-nitrophenoxy)pyridine (13.06 g,
45.6 mmol) was dissolved in methanol (228 mL) and THF (228 mL) and
cooled in an ice bath. Ammonium chloride (24.37 g, 456 mmol) was
added, followed by zinc dust (29.8 g, 456 mmol), and the mixture
was stirred in an ice bath for 30 minutes. After 30 minutes the ice
bath was removed and the reaction mixture was allowed to warm to
room temperature. After an additional hour of stirring the mixture
was filtered through Celite.RTM., which was washed well with
methanol. The filtrate was concentrated under reduced pressure and
the residue was partitioned between ethyl acetate (200 mL) and
water (100 mL). The organic layer was washed with additional water
(50 mL) and brine (100 mL), dried over magnesium sulfate, and
concentrated to give
4-(2-chloropyridin-4-yloxy)-2,5-difluorobenzenamine (11.60 g, 99%
yield) as a light brown solid. MS (ESI) m/z: 257.0 (M+H.sup.+).
Example A2
[0168] 2-Chloro-4-hydroxypyridine (0.319 g, 2.460 mmol) was
dissolved in DMF (10 mL) under argon and cooled to -15.degree. C.
Sodium hydride (60% in mineral oil) (0.148 g, 3.69 mmol) was added
slowly and the mixture was stirred for 15 minutes.
5-Chloro-2,4-difluoronitrobenzene (0.5 g, 2.58 mmol) was then added
all at once as a solution in DMF (2 mL). The reaction mixture
stirred at -15.degree. C. for 1 hour and then additional
5-chloro-2,4-difluoronitrobenzene (0.075 g) was added. The mixture
stirred at -15.degree. C. for an additional 15 hours and was then
warmed to room temperature and diluted with ethyl acetate (100 mL)
and washed with 10% aqueous lithium chloride (3.times.75 mL) and
brine (75 mL). The organic layer was dried over magnesium sulfate
and evaporated to yield an orange oil, which was then purified by
silica gel chromatography (0 to 30% ethyl acetate/hexane) to give
2-chloro-4-(2-chloro-5-fluoro-4-nitrophenoxy)pyridine (0.64 g, 86%
yield) as a light yellow oil. .sup.1H NMR (400 MHz, DMSO-d.sub.6):
.delta. 8.57 (dd, 1H), 8.36 (dd, 1H), 7.87 (dd, 1H), 7.32 (dd, 1H),
7.19 (m, 11-1); MS (ESI) m/z: 303.0 (M+H.sup.+).
[0169] 2-Chloro-4-(2-chloro-5-fluoro-4-nitrophenoxy)pyridine (0.64
g, 2.112 mmol) was dissolved in methanol (50 mL) and THF (50.0 mL).
Ammonium chloride (1.130 g, 21.12 mmol) was added, followed by zinc
dust (1.381 g, 21.12 mmol). The suspension stirred at room
temperature for 3 hours and was then filtered through Celite.RTM.
and evaporated to yield a brown solid, which was then partitioned
between a 4:1 mixture of ethyl acetate and THF (150 mL) and water
(75 mL). The organic layer was washed with brine, dried over
magnesium sulfate, and evaporated to afford
5-chloro-4-(2-chloropyridin-4-yloxy)-2-fluorobenzenamine (0.505 g,
88% yield) as a thick brown oil. MS (ESI) m/z: 273.0
(M+H.sup.+).
Example A3
[0170] A solution of 4,6-dichloro-pyrimidin-2-ylamine (5 g, 30
mmol) and acetyl chloride (4.7 g, 60 mmol) in acetic acid (200 mL)
was stirred at 80.degree. C. under nitrogen overnight. The solution
was cooled to RT and water (150 mL) was added. The mixture was
extracted with ethyl acetate (3.times.150 mL), and the combined
organics were washed with brine, dried over sodium sulfate and
concentrated to give N-(4,6-dichloro-pyrimidin-2-yl)-acetamide (5.0
g, 79% yield).
[0171] A solution of 4-amino-2,5-difluoro-phenol (3.5 g, 24 mmol),
N-(4,6-dichloro-pyrimidin-2-yl)-acetamide (5.30 g, 24 mmol) and
potassium carbonate (3.4 g, 24 mmol) in DMF (100 mL) was stirred at
50.degree. C. under nitrogen overnight. After cooling to room
temperature the reaction mixture was suspended in water (300 mL)
and extracted with ethyl acetate (3.times.200 mL). The combined
organic layers was washed with brine, dried over sodium sulfate and
concentrated. The crude product was purified by silica gel
chromatography (15%-20% ethyl acetate in petroleum ether) to give
N-[4-(4-amino-2,5-difluoro-phenoxy)-6-chloro-pyrimidin-2-yl]-acetamide
(3.3 g, 44% yield) as a white solid. .sup.1HNMR (400 MHz,
DMSO-d.sub.6): .delta. 10.72 (s, 1H), 7.20-7.24 (dd, J=11.2 Hz,
J=7.6 Hz, 1H), 7.00 (s, 1H), 6.64 (dd, 0.1=12.0 Hz, J=8.4 Hz, 1H),
5.48 (s, 2H), 2.03 (s, 3H).
[0172] A mixture of
N-[4-(4-amino-2,5-difluoro-phenoxy)-6-chloro-pyrimidin-2-yl]-acetamide
(3.3 g, 10.5 mmol) and palladium on carbon (1.0 g, 10%) in methanol
(100 mL) was stirred under H.sub.2 (1 atm) at 15.degree. C. for 4
h. The reaction mixture was filtered and the filtrate was
concentrated under reduced pressure to give
N-[4-(4-amino-2,5-difluoro-phenoxy)-pyrimidin-2-yl]-acetamide (2.4
g, 82% yield) as a pale yellow solid. .sup.1HNMR (400 MHz,
DMSO-d.sub.6): .delta. 10.37 (s, 1H), 8.44 (d, J=5.7 Hz, 1H), 7.15
(dd, J=11.4 Hz, J=4.8 Hz, 1H) 6.71 (d, J=5.7 Hz, 1H), 6.65 (dd,
J=12.3 Hz, J=8.4 Hz, 1H), 5.40 (s, 2H), 1.99 (s, 3H); MS (ESI) m/z:
281.2 [M+H].sup.+.
Example B1
[0173] Cyclopropane-1,1-dicarboxylic acid monomethylester (2 g,
13.88 mmol) was dissolved in DMF (28 mL) and 4-fluoroaniline (1.999
mL, 20.82 mmol) was added, followed by diisopropylethylamine (12.12
mL, 69.4 mmol) and
O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
tetrafluoroborate (8.91 g, 27.8 mmol). The mixture stirred at room
temperature for 15 hours and was then diluted with ethyl acetate
(200 mL) and washed with 10% aqueous lithium chloride (3.times.100
mL) and brine (100 mL). The organic layer was dried over magnesium
sulfate and evaporated to yield the crude product as a brown solid.
It was purified by silica gel chromatography (0 to 20% ethyl
acetate/hexane) to give methyl
1-((4-fluorophenyl)carbamoyl)cyclopropanecarboxylate (3.28 g, 99%
yield) as a light yellow solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6): .delta. 10.32 (s, 1H), 7.60 (m, 2H), 7.12 (m, 2H),
3.66 (s, 3H), 1.38 (m, 4H); MS (ESI) m/z: 238.1 (M+H.sup.+).
[0174] Methyl 1-((4-fluorophenyl)carbamoyl)cyclopropanecarboxylate
(3.28 g, 14.00 mmol) was dissolved in THF (23.34 mL), water (11.67
mL) was added, followed by lithium hydroxide monohydrate (1.763 g,
42.0 mmol), and the mixture stirred at room temperature for 30
minutes. After this time the THF was removed under reduced pressure
and the pH of the water layer was adjusted to .about.5 with 2 M HCl
while the solution was cooled in an ice bath. The precipitate that
formed was dissolved in ethyl acetate (125 mL) and the layers were
separated. The organic layer was washed with water (100 mL) and
brine (100 mL) and then dried over magnesium sulfate. Evaporation
of the solvent yielded
1-((4-fluorophenyl)carbamoyl)cyclopropanecarboxylic acid (2.952 g,
94% yield) as an off white powder. .sup.1H NMR (400 MHz,
DMSO-d.sub.6): .delta. 13.06 (broad s, 1H), 10.56 (s, 1H), 7.60 (m,
2H), 7.12 (m, 2H), 1.39 (s, 4H); MS (ESI) m/z: 224.1
(M+H.sup.1).
[0175] 1-((4-Fluorophenyl)carbamoyl)cyclopropanecarboxylic acid
(1.484 g, 6.65 mmol) was dissolved in thionyl chloride (14 mL, 192
mmol) at 60.degree. C. The reaction mixture stirred for 30 minutes
under argon, and then the solution was cooled to room temperature
and toluene (10 mL) was added. The mixture was concentrated under
reduced pressure. Additional toluene (10 mL) was added, and then
the mixture was again concentrated. This was repeated twice more.
The off-white solid that was obtained,
1((4-fluorophenyl)carbamoyl)cyclopropanecarbonyl chloride, was used
immediately in the next step without purification, assuming a 100%
yield. MS (ESI) m/z (methanol quench): 238.1 (M+H.sup.+).
Example B2
[0176] Cyclopropane-1,1-dicarboxylic acid monomethyl ester (0.4 g,
2.78 mmol) was, dissolved in DMF (5.55 mL) and aniline (0.380 mL,
4.16 mmol) was added, followed by diisopropylethylamine (2.424 mL,
13.88 mmol) and O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
tetrafluoroborate (1.782 g, 5.55 mmol). The reaction mixture was
stirred at room temperature for 18 hours and was then diluted with
ethyl acetate (70 mL) and washed with 10% aqueous lithium chloride
(3.times.40 mL), saturated aqueous ammonium chloride (40 mL),
saturated aqueous sodium bicarbonate (40 mL) and brine (40 mL). The
organic layer was dried over magnesium sulfate and evaporated to
yield a dark brown oil. It was purified by silica gel
chromatography (0 to 20% ethyl acetate/hexane) to yield methyl
1-(phenylcarbamoyl)cyclopropanecarboxylate (0.607 g, 100% yield) as
a light peach solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.
10.29 (s, 1H), 7.58 (d, 2H), 7.28 (t, 2H), 7.04 (t, 1H), 3.66 (s, 3
H), 1.37 (m, 4H); MS (ESI) m/z: 220.1 (M+H.sup.+).
[0177] Methyl 1-(phenylcarbamoyl)cyclopropanecarboxylate (0.607 g,
2.77 mmol) was dissolved in a mixture of THF (3.5 mL) and water
(3.50 mL), lithium hydroxide monohydrate (0.349 g, 8.31 mmol) was
added, and the mixture was stirred at room temperature for 1 hour.
The THF was removed under reduced pressure and additional water (20
mL) was added. The solution was acidified to .about.pH 4 with 2 NI
HCl and the off white solid that precipitated was collected by
suction filtration and washed with additional water to give
1-(phenylcarbamoyl)cyclopropanecarboxylic acid (0.482 g, 85%
yield). MS (ESI) m/z: 206.0 (M+H.sup.+).
[0178] 1-(Phenylcarbamoyl)cyclopropanecarboxylic acid (0.115 g,
0.559 mmol) was dissolved in thionyl chloride (1.224 mL, 16.77
mmol) and heated to 60.degree. C. under argon. After 1 hour the
reaction mixture was cooled to room temperature and evaporated to
dryness under reduced pressure. Toluene (2 mL) was added and
evaporated three times and the pale peach oil that remained,
1-(phenylcarbamoyl)cyclopropanecarbonyl chloride, was used
immediately in the next step assuming a 100% yield. MS (ESI) m/z
(methanol quench): 220.1 (M+H.sup.+).
Example 1
Compound D
[0179] Example A1 (2.136 g, 8.32 mmol) was dissolved in dry THF (63
mL) and triethylamine (1.508 mL, 10.82 mmol) was added. To this
solution was added Example B1 (2.414 g, 9.99 mmol) in dry THF (20
mL). The mixture stirred at room temperature for 30 minutes. The
triethylamine hydrochloride was removed from the reaction mixture
by suction filtration. The filtrate was evaporated to yield an
orange oil which was purified by silica gel chromatography (10% to
50% ethyl acetate/hexane) to yield
N-(4-(2-chloropyridin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluorop-
henyl)cyclopropane-1,1-dicarboxamide (3.819 g, 99% yield) as a
cream-colored solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.
11.13 (s, 1H), 9.73 (s, 1H), 8.30 (d, 1H), 8.13 (dd, 1H), 7.57 (m,
3H), 7.16 (m, 3H), 7.02 (dd, 1H), 1.64 (m, 2H), 1.57 (m, 2H); MS
(ESI) m/z: 462.1 (M+H.sup.+).
[0180]
N-(4-(2-chloropyridin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluorophen-
yl)cyclopropane-1,1-dicarboxamide (3.819 g, 8.27 mmol), acetamide
(2.442 g, 41.3 mmol), cesium carbonate (4.04 g, 12.40 mmol), and
xantphos (0.469 g, 0.810 mmol) were stirred in dry dioxane (59.1
mL) while argon was bubbled through the mixture for 15 minutes.
After this time palladium acetate (0.139 g, 0.620 mmol) was added,
and argon was bubbled through the solution for an additional 10
minutes. The round bottom flask was then fitted with a reflux
condenser, flushed with argon, and heated to 100.degree. C.
gradually from room temperature while under a balloon of argon.
After 3.5 hours at 100.degree. C. the reaction mixture was cooled
to room temperature. The reaction mixture was diluted with a 4:1
mixture of ethyl acetate and THF (300 mL) and water (100 mL). A
bright yellow solid was removed by suction filtration and
discarded. The organic layer was separated from the aqueous and
washed with brine (200 mL). In addition, the aqueous layer was
back-extracted with the ethyl acetate/THF mixture (100 mL), which
was then also washed with brine (50 mL). The combined organic
layers were dried over magnesium sulfate and evaporated to yield a
peach-colored oil. It was stirred in dichloromethane (50 mL) for
1.5 hours, and a white solid formed which was collected by suction
filtration and washed with more dichloromethane to give
N-(4-(2-acetamidopyridin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluorophenyl)-
cyclopropane-1,1-dicarboxamide (3.328 g, 83% yield). .sup.1H NMR
(400 MHz, DMSO-d.sub.6): .delta. 11.00 (s, 1H), 10.59 (s, 1H), 9.79
(s, 1H), 8.19 (d, 1H), 8.07 (dd, 1H), 7.65 (d, 1H), 7.57 (m, 3H),
7.16 (m, 2H), 6.71 (dd, 1H), 2.02 (s, 3H), 1.62 (m, 2H), 1.57 (m,
2H); MS (ESI) m/z: 485.1 (M+H.sup.+).
Example 2
[0181] A solution of Example A2 (0.147 g, 0.538 mmol) in dry THF
(5.38 mL) with triethylamine (0.098 mL, 0.700 mmol) was added to
Example B1 (0.169 g, 0.699 mmol). The mixture was stirred under
argon for 20 minutes at room temperature. The reaction mixture was
then filtered through a frit in order to remove the solid
triethylamine hydrochloride that had precipitated. The filtrate was
concentrated under reduced pressure to yield a pale orange oil
which was purified by silica gel chromatography (0 to 50% ethyl
acetate/hexane) to yield
N-(5-chloro-4-(2-chloropyridin-4-yloxy)-2-fluorophenyl)-N'-(4-fluoropheny-
l)cyclopropane-1,1-dicarboxamide (0.203 g, 79% yield) as a clear
sticky oil. .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 11.04 (s,
1H), 9.77 (s, 1H), 8.30 (m, 2H), 7.58 (m, 3H), 7.16 (t, 2H), 7.07
(d, 1H), 6.96 (dd, 1H), 1.63 (m, 2H), 1.56 (m, 2H); MS (ESI) m/z:
478.1 (M+H.sup.+).
[0182]
N-(5-chloro-4-(2-chloropyridin-4-yloxy)-2-fluorophenyl)-N'-(4-fluor-
ophenyl)cyclopropane-1,1-dicarboxamide (0.200 g, 0.418 mmol),
acetamide (0.124 g, 2.091 mmol), cesium carbonate (0.136 g, 0.418
mmol), and xantphos (0.017 g, 0.029 mmol) were dissolved in dry
dioxane (3 mL) in a 25 mL round bottom flask. Argon was bubbled
through the reaction mixture for 5 minutes, and then palladium
acetate (4.69 mg, 0.021 mmol) was added. The mixture was again
degassed for five minutes, and then the reaction flask was fitted
with a reflux condenser. The system was flushed with argon and then
heated at 100.degree. C. under a balloon of argon for 3 hours. The
reaction mixture was cooled to room temperature and diluted with
water (30 mL) and a 4:1 mixture of ethyl acetate and THF (150 mL).
The layers were separated, and the aqueous layer was washed with
additional ethyl acetate/THF solution. The organic layers were
combined and concentrated to yield a sticky orange oil. Upon
addition of methanol (3 mL) a fine cream-colored precipitate
formed, which was collected by suction filtration and washed with a
small portion of dichloromethane to yield
N-(4-(2-acetamidopyridin-4-yloxy)-5-chloro-2-fluorophenyl)-N'-(4-fl-
uorophenyl)cyclopropane-1,1-dicarboxamide (0.100 g, 47.7% yield).
.sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 10.91 (s, 1H), 10.58
(s, 1H), 9.83 (s, 1H), 8.23 (d, 1H), 8.18 (d, 1H), 7.58 (m, 4H),
7.15 (m, 2H), 6.65 (dd, 1H), 2.02 (s, 3H), 1.61 (m, 2H), 1.56 (m,
2H); MS (ESI) m/z: 501.1 (M+H.sup.+).
Example 3
[0183] Example A 1 (0.12 g, 0.468 mmol) was dissolved in dry THF
(4.68 mL) and triethylamine (0.085 mL, 0.608 mmol) was added. This
solution was added to Example 132 (0.125 g, 0.561 mmol) and the
mixture stirred under argon at room temperature for 2 hours. The
reaction mixture was filtered to remove triethylamine hydrochloride
salt and the filtrate was evaporated to yield a light peach oil
which was purified by silica gel chromatography (10 to 50% ethyl
acetate/hexane) to yield
N-(4-(2-chloropyridin-4-yloxy)-2,5-difluorophenyl)-N'-phenylcyclopropane--
1,1-dicarboxamide (0.164 g, 79% yield) as a clear solid. .sup.1H
NMR (400 MHz, DMSO-d.sub.4): .delta. 11.10 (s, 1H), 9.71 (s, 1H),
8.31 (d, 1H), 8.12 (dd, 1H), 7.60 (dd, 1H), 7.55 (m, 2H), 7.32 (t,
2H), 7.14 (d, 1H), 7.10 (m, 1H), 7.02 (dd, 1H), 1.65 (m, 2H), 1.58
(m, 2H); MS (ESI) m/z: 444.1 (M+H.sup.+).
[0184]
N-(4-(2-chloropyridin-4-yloxy)-2,5-difluorophenyl)-N'-phenylcyclopr-
opane-1,1-dicarboxamide (0.162 g, 0.365 mmol), acetamide (0.108 g,
1.825 mmol), cesium carbonate (0.178 g, 0.548 mmol), and xantphos
(0.021 g, 0.036 mmol) were combined in dry dioxane (2.61 mL) and
argon was bubbled through the mixture for 5 minutes. Palladium
acetate (6.15 mg, 0.027 mmol) was added, and argon was bubbled
through the mixture for an additional 5 minutes. The reaction flask
was fitted with a reflux condenser and a balloon of argon and the
mixture was heated at 100.degree. C. for 20 hours. The reaction
mixture was cooled to room temperature and partitioned between a
4:1 mixture of ethyl acetate and THF (50 mL) and water (50 mL). The
aqueous layer was removed and the organic layer was washed with
additional water (50 mL) and brine (50 mL). The organic layer was
dried over magnesium sulfate and evaporated under reduced pressure
to yield a light peach-colored film. Dichloromethane (10 mL) was
added and after a few minutes solid began to precipitate.
Sonication was used to precipitate out more solid. After sitting
for 30 minutes the bright white solid was collected by suction
filtration and washed with additional dichloromethane to give
N-(4-(2-acetamidopyridin-4-yloxy)-2,5-difluorophenyl)-N'-phenylcyclopropa-
ne-1,1-dicarboxamide (0.099 g, 58% yield). .sup.1H NMR (400 MHz,
DMSO-d.sub.6): .delta. 10.98 (s, 1H), 10.59 (s, 1H), 9.78 (s, 1H),
8.19 (d, 1H), 8.07 (dd, 1H), 7.65 (d, 1H), 7.55 (m, 3H), 7.32 (m,
2H), 7.09 (t, 1H), 6.71 (dd, 1H), 2.02 (s, 3H), 1.63 (m, 2H), 1.57
(m, 2H); MS (ESI) m/z: 467.2 (M+H.sup.+).
Example 4
[0185] 2-Bromoacetamide (1 g, 7.25 mmol) was dissolved in
acetonitrile (10.35 mL) and 2 M dimethylamine in THF (12 mL, 24.00
mmol) was added. The mixture stirred under argon at room
temperature for 48 hours. The reaction mixture was evaporated under
reduced pressure and the residue was re-dissolved in a 1:1 mixture
of dichloromethane and methanol (50 mL). It was neutralized over a
carbonate resin (2 equiv) with gentle shaking for 20 hours. The
reaction mixture was filtered and the filtrate was evaporated to
yield 2-(dimethylamino)acetamide (0.740 g, 100% yield) as a
peach-colored solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.
7.34 (s, 1H), 7.19 (s, 1H), 3.01 (s, 2H), 2.31 (s, 6H).
[0186] 2-(Dimethylamino)acetamide (0.100 g, 0.974 mmol),
N-(4-(2-chloropyridin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluorophenyl)cyc-
lopropane-1,1-dicarboxamide (0.15 g, 0.325 mmol) (as prepared in
Example 1), cesium carbonate (0.159 g, 0.487 mmol), and xantphos
(0.018 g, 0.032 mmol) were combined in dry dioxane (2.5 mL) and
argon was bubbled through the mixture for 5 minutes. Palladium
acetate (5.47 mg, 0.024 mmol) was added and the solution was
degassed for an additional 5 minutes. The reaction flask was fitted
with a reflux condenser and a balloon of argon and heated at
100.degree. C. for 15 hours. The mixture was cooled to room
temperature and then diluted with ethyl acetate (75 mL) and water
(45 mL). The water layer was removed and extracted again with ethyl
acetate (25 mL). The combined organic layers were washed with brine
(50 mL) and dried over magnesium sulfate. Evaporation of the
solvent yielded a lavender-colored oil which was purified by silica
gel chromatography (0 to 7% methanol in dichloromethane) to give
N-(4-(2-(2-(dimethylamino)acetamido)pyridin-4-yloxy)-2,5-difluorophenyl)--
N'-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide (0.0823 g, 48%
yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 11.02 (s, 1H),
9.97 (s, 1H), 9.79 (s, 1H), 8.20 (d, 1H), 8.09 (dd, 1H), 7.65 (d,
1H), 7.57 (m, 3H), 7.16 (m, 2H), 6.76 (dd, 1H), 3.06 (s, 2H), 2.25
(s, 6H), 1.63 (m, 2H), 1.57 (m, 2H); MS (ESI) m/z: 528.2
(M+H.sup.+).
Example 5
[0187] To a solution of Example A3 (300 mg, 1.07 mmol) and
1-(4-fluoro-phenylcarbamoyl)-cyclopropanecarboxylic acid (240 mg,
1.07 mmol) (as prepared in Example B1) in DMF (20 mL) was added
HATU (440 mg, 3.2 mmol) and DIEA (280 mg, 2.1 mmol) in portions.
The reaction mixture was stirred at 60.degree. C. under nitrogen
overnight. After cooling to room temperature water (30 mL) was
added and the solution was extracted with ethyl acetate (3.times.50
mL). The combined organics were washed with brine (3.times.50 mL),
dried over sodium sulfate, and concentrated. The crude product was
purified by preparative HPLC to give
N-(4-(2-acetamidopyrimidin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluoropheny-
l)cyclopropane-1,1-dicarboxamide (42 mg, 8% yield) as a white
solid. .sup.1HNMR (300 MHz, DMSO-d.sub.6): .delta. 10.95 (s, 1H),
10.40 (s, 1H), 9.75 (s, 1H), 8.49-8.51 (d, J=5.7 Hz, 1H), 7.94-8.01
(dd, J=12.3 Hz, J=8.1 Hz, 1H), 7.48-7.60 (m, 3H), 7.05-7.16 (m,
2H), 6.83-6.84 (d, J=5.4 Hz, 1H), 1.92 (s, 3H), 1.53-1.59 (d,
J=19.2 Hz, 4H).
Example 6
[0188]
N-(4-(2-Chloropyridin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluorophen-
yl)cyclopropane-1,1-dicarboxamide (0.25 g, 0.541 mmol) (as prepared
in Example 1), cyclopropanecarboxamide (0.092 g, 1.083 mmol),
xantphos (0.014 g, 0.024 mmol), and cesium carbonate (0.265 g,
0.812 mmol) were dissolved in dry dioxane (5.41 mL) and argon was
bubbled through the mixture for 5 minutes. Pd.sub.2(dba).sub.3
(7.44 mg, 0.00812 mmol) was added and additional argon was bubbled
through the system. It was then fitted with a reflux condenser and
a balloon of argon and heated at 100.degree. C. for 20 hours. The
reaction mixture was cooled to room temperature and then
partitioned between water (40 mL) and ethyl acetate (70 mL). The
layers were separated and the organic layer was washed with brine
(50 mL), dried over magnesium sulfate, and evaporated to yield a
peach-colored solid. It was stirred in dichloromethane (10 mL) and
a cream-colored solid was collected by suction filtration and
washed with additional dichloromethane to yield
N-(4-(2-(cyclopropanecarboxamido)pyridin-4-yloxy)-2,5-difluorophenyl)-N'--
(4-fluorophenyl)cyclopropane-1,1-dicarboxamide (0.238 g, 86%
yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 11.00 (s, 1H),
10.89 (s, 1H), 9.79 (s, 1H), 8.20 (d, 1H), 8.07 (dd, 1H), 7.63 (d,
1H), 7.56 (m, 3H), 7.16 (m, 2H), 6.74 (dd, 1H), 1.95 (quintet, 1H),
1.62 (m, 2H), 1.56 (m, 2H), 0.75 (m, 4H); MS (ESI) m/z: 511.1
(M+H.sup.+).
Example 7
[0189]
N-(4-(2-Chloropyridin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluorophen-
yl)cyclopropane-1,1-dicarboxamide (200 mg, 0.43 mmol) (as prepared
in Example 1), propionamide (95 mg, 1.30 mmol), xantphos (25 mg,
0.043 mmol), and cesium carbonate (280 mg, 0.86 mmol) were
dissolved in dry dioxane (3 mL) and argon was bubbled through the
mixture for 10 minutes. Pd.sub.2(dba).sub.3 (20 mg, 0.022 mmol) was
then added, and the solution was degassed for an additional 10
minutes. The flask was fitted with a balloon of N.sub.2 and slowly
heated to 100.degree. C. and stirred overnight. The reaction
mixture was cooled to room temperature and diluted with a 4:1
mixture of ethyl acetate and THF (60 mL) and water (40 mL). The
organic layer was separated and washed with brine and the aqueous
layer was back extracted with the ethyl acetate/THF solution, which
was then extracted with brine. The combined organic layers were
dried over sodium sulfate and evaporated and the residue was
purified by silica gel chromatography to give
N-(2,5-difluoro-4-(2-propionamidopyridin-4-yloxy)phenyl)-N'-(4-fluorophen-
yl)cyclopropane-1,1-dicarboxamide (130 mg, 60.7% yield).
.sup.1H-NMR (400 MHz, DMSO-d.sub.6): .delta. 11.00 (s, 1H), 10.52
(s, 1H), 9.79 (s, 1H), 8.19 (d, J=5.6 Hz, 1H), 8.10-8.05 (m, 1H),
7.66 (d, J=2.4 Hz, 1H), 7.59-7.53 (m, 3H), 7.16 (t, J=8.8 Hz, 2H),
6.74-6.72 (m, 1H), 2.33 (q, J=7.2 Hz, 2H), 1.64-1.55 (m, 4H), 0.99
(t, J=7.2 Hz, 3H).
Example 8
Reference Compound E
[0190] 1-((4-Fluorophenyl)carbamoyl)cyclopropanecarboxylic acid
(see Example B1, 171 mg, 0.77 mmol),
N-(4-methoxybenzyl)-4-(4-amino-3-fluorophenoxy)pyridin-2-amine (see
PCT Publication No. WO 2008/046003, 200 mg, 0.59 mmol), TBTU (284
mg, 0.88 mol) and DIEA (0.12 mL, 0.73 mmol) were combined in DMF
(1.5 mL) and the resultant mixture was stirred overnight. The
reaction mixture was partitioned between saturated aqueous
NaHCO.sub.3 (20 mL) and EtOAc (20 mL). The organic was washed with
water (10 mL), brine (10 mL), and 5% aqueous lithium chloride
solution (10 mL), and was then dried over MgSO.sub.4 and
concentrated in vacuo. Dichloromethane was added to the residue and
the resultant slurry was filtered. The collected precipitate was
washed with CH.sub.2Cl.sub.2 and dried in vacuo to provide
N-(4-(2-(4-methoxybenzylamino)pyridin-4-yloxy)-2-fluorophenyl)-N'-(4-fluo-
rophenyl)cyclopropane-1,1-dicarboxamide (137 mg) as a white solid.
The filtrate was concentrated and a second crop (46 mg, total yield
57%) was collected. MS (ESI) m/z: 545.1 (M+H.sup.+).
[0191] A mixture of
N-(4-(2-(4-methoxybenzylamino)pyridin-4-yloxy)-2-fluorophenyl)-N'-(4-fluo-
rophenyl)cyclopropane-1,1-dicarboxamide (160 mg, 0.29 mmol) in
CH.sub.2Cl.sub.2 (0.2 mL) was treated with trifluoroacetic acid
(0.4 mL, 5.26 mmol) and the resultant mixture was stirred overnight
at RT. The reaction mixture was concentrated to dryness and the
residue was purified by reverse-phase silica gel chromatography
(25-95% acetonitrile in water, 0.1% TFA). The desired fractions
were partitioned between saturated aqueous NaHCO.sub.3 and EtOAc.
The organics were washed with saturated aqueous NaHCO.sub.3, water,
and brine and were dried over Na.sub.2SO.sub.4 and concentrated in
vacuo to provide
N-(4-(2-aminopyridin-4-yloxy)-2-fluorophenyl)-N'-(4-fluorophenyl)cyclopro-
pane-1,1-dicarboxamide (52 mg, 39% yield). MS (ESI) m/z: 425.1
(M+H.sup.+).
[0192] A solution of
N-(4-(2-aminopyridin-4-yloxy)-2-fluorophenyl)-N'-(4-fluorophenyl)cyclopro-
pane-1,1-dicarboxamide (61 mg, 0.14 mmol) in CH.sub.2Cl.sub.2 (3
mL) was treated with pyridine (0.058 mL, 0.72 mmol) and acetic
anhydride (0.13 mL, 1.4 mmol). The resultant mixture was stirred at
RT for 2 days. The reaction was quenched with saturated aqueous
NaHCO.sub.3 and was further stirred for 2 h. The mixture was
diluted with EtOAc (30 mL) and was washed with saturated aqueous
NaHCO.sub.3 (20 mL), water (20 mL) and brine (20 mL). The mixture
was concentrated in vacuo and purified by silica gel chromatography
to provide
N-(4-(2-acetamidopyridin-4-yloxy)-2-fluorophenyl)-N'-(4-fluorophenyl)cycl-
opropane-1,1-dicarboxamide (46 mg, 69% yield). .sup.1H NMR (400
MHz, DMSO-d.sub.6): .delta.10.56 (s, 1H), 10.54 (s, 1H), 9.96 (s,
1H), 8.18 (d, J=5.8 Hz, 1H), 7.92 (t, 1H), 7.65 (d, J=1.9 Hz, 1H),
7.59 (m, 2H), 7.24 (dd, J=11.2, 2.5 Hz, 1H), 7.15 (m, 2H), 7.01 (m,
1H), 6.68 (dd, J=5.7, 2.3 Hz, 1H), 2.02 (s, 3H), 1.60-1.52 (m, 4H);
MS (ESI) m/z: 467.2 (M+H.sup.+).
Example 9
[0193]
N-(4-(2-Chloropyridin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluorophen-
yl)cyclopropane-1,1-dicarboxamide (3 g, 6.5 mmol) (as prepared in
Example 1), tert-butyl carbamate (2.3 g, 19.5 mmol), Xantphos (0.37
g, 0.65 mmol), and cesium carbonate (4.2 g, 13 mmol) were dissolved
in dry dioxane (50 mL) and argon was bubbled through the mixture
for 10 minutes. Pd.sub.2(dba).sub.3 (0.3 g, 0.33 mmol) was then
added, and the solution was sparged with argon for an additional 10
minutes. The flask was fitted with a reflux condenser and a balloon
of argon and slowly heated to 100.degree. C. and stirred overnight.
The reaction mixture was cooled to RT. It was diluted with ethyl
acetate (100 mL) and water (80 mL). The organic layer was separated
and washed with brine. The aqueous layer was back extracted with
the ethyl acetate, which was then washed with brine. The combined
organic layers were dried over sodium sulfate and evaporated. The
residue was purified by silica gel chromatography to give
tert-butyl
difluoro-4-(1-(4-fluorophenyl)carbamoyl)cyclopropanecarboxamido)phenoxy)p-
yridin-2-ylcarbamate (1.8 g, 51% yield). .sup.1H NMR (400 MHz,
DMSO-d.sub.6): .delta. 11.03 (s, 1H), 9.88 (s, 1H), 9.78 (s, 1H),
8.13-8.05 (m, 2H), 7.59-7.53 (m, 3H), 7.33 (d, J=2.4 Hz, 1H),
7.18-7.13 (m, 2H), 6.63 (d, J=2.4 Hz, 1H), 1.63-1.62 (m, 2H),
1.58-1.56 (m, 2H), 1.40 (s, 9H).
[0194] To a solution of tert-butyl
4-(2,5-difluoro-4-(1-(4-fluorophenylcarbamoyl)cyclopropanecarboxamido)phe-
noxy)pyridin-2-ylcarbamate (1.8 g, 3.3 mmol) in CH.sub.2Cl.sub.2
(100 mL) was added TPA (5 mL) and the mixture was stirred at room
temperature overnight. The reaction mixture was adjusted to pH>7
with saturated NaHCO.sub.3 solution and the separated organic layer
was washed with brine, dried over Na.sub.2SO.sub.4 and concentrated
under reduced pressure to give
N-(4-((2-aminopyridin-4-yl)oxy)-2,5-difluorophenyl)-N'-(4-fluorophenyl)cy-
clopropane-1,1-dicarboxamide (1.2 g, yield 82% yield). .sup.1H-NMR
(400 MHz, DMSO-d.sub.6): .delta. 10.99 (s, 1H), 9.76 (s, 1H), 8.04
(dd, J=12.4, 7.6 Hz, 1H), 7.79 (d, J=6.0 Hz, 1H), 7.58-7.55 (m,
2H), 7.47 (dd, J=10.8, 7.2 Hz, 1H), 7.16 (t, J=8.8 Hz, 2H), 6.16
(dd, J=6.0, 2.4 Hz, 1H), 6.00 (br s, 2H), 5.81 (d, J=2.0 Hz, 1H),
1.65-1.62 (m, 2H), 1.56-1.53 (m, 2H); MS (ESI): m/z 443.1
[M+H].sup.+.
[0195] To a solution of
N-(4-((2-aminopyridin-4-yl)oxy)-2,5-difluorophenyl)-N'-(4-fluorophenyl)cy-
clopropane-1,1-dicarboxamide (130 mg, 0.29 mmol) in 10 mL of
anhydrous tetrahydrofuran was added diisopropylethylamine (75 mg,
0.58 mmol). A solution of methoxyacetyl chloride (34.5 mg, 0.32
mmol) in THF (1.degree. mL) was added drop wise at 0.degree. C. The
resultant reaction mixture was stirred at R.T. for 0.5 h. It was
diluted with ethyl acetate, washed with brine, dried over sodium
sulfate and concentrated. The residue was purified by prep-TLC to
give
N-(2,5-difluoro-4-((2-(2-methoxyacetamido)pyridin-4-yl)oxy)phenyl)-N'-(4--
fluorophenyl)cyclopropane-1,1-dicarboxamide (75 mg, 50% yield).
.sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 9.79 (s, 1H), 8.89 (s,
1H), 8.53 (s, 1H), 8.30 (dd, J=12.0, 7.2 Hz, 1H), 8.17 (d, J=5.6
Hz, 1H), 7.83 (d, J=2.4 Hz, 1H), 7.49-7.45 (m, 2H), 7.07-6.99 (m,
3H), 6.64 (dd, J=5.6, 2.4 Hz, 1H), 3.98 (s, 2H), 3.49 (s, 3H),
1.78-1.66 (m, 4H); MS (ESI): m/z 515.2 [M+H].sup.+.
Example 10
[0196] In degassed dioxane (5 mL) was placed
N-(4-(2-chloropyridin-4-yloxy)-2,5-difluorophenyl)-N-(4-fluorophenyl)cycl-
opropane-1,1-dicarboxamide (250 mg, 0541 mmol) (as prepared in
Example 1), cesium carbonate (353 mg, 1.083 mmol), isobutrylamide
(236 mg, 2.71 mmol) and xantphos (31 mg, 54 .mu.mol). To this was
added tris(dibenzylideneacetone)dipalladium(0) (25 mg, 27 .mu.mol).
The mix was warmed to 100.degree. C. overnight. The mixture was
cooled to room temperature and diluted with ethyl acetate (30 mL)
and filtered to remove solids. The filtrate was washed with aq
NaHCO.sub.3 (30 mL) and brine (30 mL). The organic phase was dried
over Na.sub.2SO.sub.4 and evaporated at reduced pressure to give a
foam. The foam was purified by reverse phase silica gel
chromatography (35-80% acetonitrile/water/0.1% TFA). Fractions
containing product were combined and evaporated at reduced
pressure. The resultant aqueous mixture was then treated with
saturated aq NaHCO.sub.3 (4 mL) and allowed to stand. The solid was
collected by filtration, washed with water (2.times.5 mL) and dried
on high vacuum line at 80.degree. C. overnight to provide
N-(2,5-difluoro-4-(2-isobutyramidopyridin-4-yloxy)phenyl)-N'-(4-fluorophe-
nyl)cyclopropane-1,1-dicarboxamide (116 mg, 41% yield). .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 11.00 (s, 1H), 10.53 (s, 1H), 9.78
(s, 1H), 8.19 (d, 1H), 8.06-8.10 (m, 1H), 7.66 (s, 1H), 7.60-7.53
(m, 3H), 7.15 (t, 2H), 6.75-6.73 (m, 1H), 2.68 (m, 1H), 1.62-1.51
(m, 4H), 1.00 (d, 6H); MS (ES-API) m/z: 513.2 (M+H.sup.+).
Example 11
[0197] To a solution of
N-(4-((2-aminopyridin-4-yl)oxy)-2,5-difluorophenyl)-N'-(4-fluorophenyl)cy-
clopropane-1,1-dicarboxamide (150 mg, 0.34 mmol) (as prepared in
Example 9) and 2-cyanoacetic acid (44 mg, 0.51 mmol) in DMF (2 mL)
was added HATU (258 mg, 0.68 mmol) and DIEA (130 mg, 1 mmol) and
the mixture was stirred at 60.degree. C. under nitrogen overnight.
The reaction mixture was cooled to room temperature, diluted with
ethyl acetate (50 mL) and H.sub.2O (50 mL) and the organic layer
was washed with brine, dried and concentrated under reduced
pressure. The residue was purified by prep-TLC to give
N-(4-(2-(2-cyanoacetamido)pyridin-4-yloxy)-2,5-difluorophenyl)-N--
(4-fluorophenyl)cyclopropane-1,1-dicarboxamide (70 mg, yield
64.5%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6): 10.99 (s, 1H), 10.94
(s, 1H), 9.81 (s, 1H), 8.23 (d, J=5.6 Hz, 1H), 8.08 (dd, J=12.4,
6.8 Hz, 1H), 7.59-7.55 (m, 4H), 7.15 (t, J=8.8 Hz, 2H), 6.80 (dd,
J=6.0, 2.4 Hz, 1H), 3.92 (s, 2H), 1.61-1.56 (m, 4H); MS (ESI): m/z
510.2 [M+H].sup.+.
Example 12
[0198] To a solution of
N-(4-((2-aminopyridin-4-yl)oxy)-2,5-difluorophenyl)-N'-(4-fluorophenyl)cy-
clopropane-1,1-dicarboxamide (440 mg, 1 mmol) (as prepared in
Example 9) and 1-(tert-butoxycarbonyl)azetidine-3-carboxylic acid
(402 mg, 2 mmol) in DMF (5 mL) was added HATU (1.1 g, 3 mmol),
followed by DIEA (516 mg, 4 mmol). The mixture was sparged with
nitrogen and then stirred 50.degree. C. overnight. The reaction
mixture was partitioned between ethyl acetate and water. The
organic layer was washed with brine, dried over sodium sulfate and
concentrated under reduced pressure. The residue was purified by
column on a silica gel chromatography to give tert-butyl
3-(4-(2,5-difluoro-4-(1-(4-fluorophenylcarbamoyl)cyclopropanecarboxamido)-
phenoxy)pyridin-2-ylcarbamoyl)azetidine-1-carboxylate (250 mg, 40%
yield). .sup.1H-NMR (400 MHz, DMSO-d.sub.6): 11.01 (s, 1H), 10.72
(s, 1H), 9.79 (s, 1H), 8.19 (d, J=5.6 Hz, 1H), 8.10-8.05 (m, 1H),
7.66 (s, 1H), 7.58-7.53 (m, 3H), 7.14 (t, J=8.8 Hz, 2H), 6.77-6.75
(m; 1H), 3.90-3.84 (m, 4H), 3.55-5.53 (m, 1H), 1.63-1.53 (m, 4H),
1.33 (s, 9H).
[0199] To a solution of tert-butyl
3-(4-(2,5-difluoro-4-(1-(4-fluorophenylcarbamoyl)cyclopropanecarboxamido)-
phenoxy)pyridin-2-ylcarbamoyl)azetidine-1-carboxylate (220 mg, 0.35
mmol) in CH.sub.2Cl.sub.2 (4 mL) was added TFA (0.2 mL) at
0.degree. C. and the reaction was stirred at room temperature
overnight. Saturated NaHCO.sub.3 solution was added drop wise until
pH>7 and the mixture with extracted with CH.sub.2Cl.sub.2
(2.times.50 mL). The combined organic layers were washed with
brine, dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The residue was purified by HPLC separation to
give
N-(4-(2-(azetidine-3-carboxamido)pyridin-4-yloxy)-2,5-difluorophenyl-
)-N'-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide (16 mg, yield
8.7%). .sup.1H-NMR (400 MHz, MeOH-d.sub.4): .delta. 9.50 (s), 8.46
(d, J=7.2 Hz, 1H), 8.33 (dd, J=7.2 Hz, 12.4 Hz, 1H), 7.53-7.43 (m,
3H), 7.26 (dd, J=7.2 Hz, 2.4 Hz, 1H), 7.10 (t, J=8.4 Hz, 2H), 6.75
(d, J=2.4 Hz, 1H), 4.61-4.54 (m, 1H), 3.63-3.58 (m, 1H), 3.49-3.44
(m, 1H), 3.31-3.29 (m, 1H), 3.27-3.25 (m, 1H), 1.75-1.73 (m, 4H);
MS (ESI): m/z 443.2 [M+H].sup.+.
Example 13
[0200] To a solution of
N-(4-(2-chloropyridin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluorophenyl)cyc-
lopropane-1,1-dicarboxamide (200 mg, 0.45 mmol) (as prepared in
Example 9) and cyclobutanecarboxylic acid (90 mg, 0.9 mmol) in DMF
(3 mL) was added HATU (513 mg, 1.35 mmol), followed by DIEA (516
mg, 4 mmol). The mixture was sparged with nitrogen and stirred
overnight at 60.degree. C. The reaction mixture was partitioned
between ethyl acetate and water. The organic layer was washed with
brine, dried over sodium sulfate and concentrated under reduced
pressure. The residue was purified by silica gel chromatography to
give
N-(4-(2-(cyclobutanecarboxamido)pyridin-4-yloxy)-2,5-difluorophenyl)-N'-(-
4-fluorophenyl)cyclopropane-1,1-dicarboxamide (97 mg, yield 41.1%).
.sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 11.02 (s, 1H), 10.42
(s, 1H), 9.80 (s, 1H), 8.18 (d, J=6.0 Hz, 1H), 8.08 (dd, J=12.4,
6.8 Hz, 1H), 7.68 (s, 1H), 7.60-7.54 (m, 3H), 7.17 (t, J=8.8 Hz,
2H), 6.73 (dd, J=5.6, 2.4 Hz, 1H), 2.18-2.02 (m, 4H), 1.92-1.83 (m,
1H), 1.76-7.69 (m, 1H), 1.62-1.55 (m, 4H); MS (ESI): m/z 525.1
[M+H]
Example 14
[0201] To a solution of (R)-2-methoxy-propionic acid (300 mg, 2.88
mmol) and N-methylmorpholine (432 mg, 4.32 mmol) in anhydrous
CH.sub.2Cl.sub.2 was added drop wise isobutyl chloroformate (588
mg, 4.32 mmol). The resultant mixture was stirred at RT for 1 h.
N-(4-(2-Chloropyridin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluorophenyl)cyc-
lopropane-1,1-dicarboxamide (200 mg, 0.452 mmol) was added and the
resulting mixture was stirred at RT for 12 h. The reaction mixture
was concentrated under vacuo and purified by HPLC separation to
give
(R)--N-(2,5-difluoro-4-((2-(2-methoxypropanamido)pyridin-4-yl)oxy)phenyl)-
-N'-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide (50 mg, 21%).
.sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 9.79 (s, 1H), 9.07 (s,
1H), 8.51 (s, 1H), 8.32-8.27 (m, 1H), 8.16 (d, J=5.6 Hz, 1H), 7.83
(d, J=2 Hz, 1H), 7.48-7.44 (m, 2H), 7.06-6.98 (m, 3H), 6.66-6.64
(m, 1H), 3.82 (q, J=6.8 Hz, 1H), 3.50 (s, 3H), 1.75-1.67 (m, 4H),
1.42 (d, J=6.8 Hz, 3H); MS (ESI): m/s 529.1 [M+H].sup.+.
Example 15
[0202] To a solution of (R)-2-benzyloxy-propionic acid (500 mg,
2.78 mmol) and N-methylmorpholine (416 mg, 4.16 mmol) in anhydrous
CH.sub.2Cl.sub.2 was added isobutyl chloroformate (566 mg, 4.32
mmol) dropwise. The mixture was stirred at 25.degree. C. for 1 h.
N-(4-((2-aminopyridin-4-yl)oxy)-2,5-difluorophenyl)-N'-(4-fluorophenyl)cy-
clopropane-1,1-dicarboxamide (200 mg, 0.452 mmol) was added and the
resulting mixture was stirred at 25.degree. C. for 12 h. The
reaction mixture was concentrated in vacuo to give
(R)--N-(4-((2-(2-(benzyloxy)propanamido)pyridin-4-yl)oxy)-2,5-difluorophe-
nyl)-N'-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide (200 mg,
71.5%). It was used without further purification.
[0203] To a solution of
(R)--N-(4-((2-(2-(benzyloxy)propanamido)pyridin-4-yl)oxy)-2,5-difluorophe-
nyl)-N'-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide (200 mg,
0.331 mmol) in methanol (20 mL) was added Pd(OH).sub.2 (50 mg). The
mixture was then hydrogenated (1 atm) for 12 h. The reaction
mixture was filtered, concentrated and purified by HPLC
chromatography to give
(R)--N-(2,5-difluoro-4-((2-(2-hydroxypropanamido)pyridin-4-yl)oxy)phenyl)-
-N'-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide (20 mg, 11.7%
yield). .sup.1H NMR (400 MHz, CD.sub.3OD): .delta. 8.23-8.18 (m,
2H), 7.69 (s, 1H), 7.54-7.50 (m, 2H), 7.31-7.28 (m, 1H), 7.09-7.05
(m, 2H), 6.86-6.84 (m, 1H), 4.28-4.23 (q, J=6.8 Hz, 1H), 1.76-1.67
(m, 4H), 1.39 (d, J=6.8 Hz, 3H); MS (ESI); m/z 515.1
[M+H].sup.+
Example 16
[0204]
N-(4-(2-chloropyridin-4-yloxy)-2,5-difluorophenyl)-N'-(4-fluorophen-
yl)cyclopropane-1,1-dicarboxamide (160 mg, 0.34 mmol) (as prepared
in Example 1), 2,2-dimethyl-propionamide (100 mg, 1 mmol), xantphos
(40 mg, 0.068 mmol), and cesium carbonate (222 mg, 0.68 mmol) were
dissolved in dry dioxane (2 mL) and argon was bubbled through the
mixture for 10 minutes. Pd(OAc).sub.2 (8 mg, 0.034 mmol) was then
added, and the solution was degassed for an additional 10 minutes.
The mixture was heated to 100.degree. C. and stirred overnight. The
reaction mixture was cooled to RT and diluted with EtOAc (20 mL)
and water (15 mL). The organic layer was separated and washed with
brine. The aqueous layer was extracted with EtOAc and the extracts
were washed with brine. The combined organic layers were dried over
sodium sulfate and concentrated in vacuo. The residue was purified
by prep-TLC to give
N-(2,5-difluoro-4-((2-pivalamidopyridin-4-yl)oxy)phenyl)-N'-(4-fluorophen-
yl)cyclopropane-1,1-dicarboxamide (72 mg, 40% yield). .sup.1H-NMR
(400 MHz, DMSO-d.sub.6): .delta. 8.19-8.17 (m, 2H), 7.69 (s, 1H),
7.53-7.51 (m, 2H), 7.24 (br t, J=7.2 Hz, 1H), 7.10-7.05 (m, 2H),
6.73 (brs, 1H), 1.73-1.69 (m, 4H), 1.27 (s, 9H); MS (ESI); m/z
527.3 [M+H].sup.+.
Example 17
[0205] To a solution of (S)-2-methoxy-propionic acid (400 mg, 3.84
mmol) and N-methylmorpholine (577 mg, 5.77 mmol) in anhydrous
CH.sub.2Cl.sub.2 was added drop wise isobutyl chloroformate (773
mg, 5.77 mmol) and the resultant mixture was stirred at 25.degree.
C. for 1 h.
N-(4-((2-aminopyridin-4-yl)oxy)-2,5-difluorophenyl)-N'-(4-fluorophenyl)cy-
clopropane-1,1-dicarboxamide (200 mg, 0.452 mmol) was added to the
mixture. The result mixture was stirred at 25.degree. C. for 12 h.
The reaction mixture was concentrated under reduced pressure and
purified by HPLC chromatography to give
(S)--N-(2,5-difluoro-4-((2-(2-methoxypropanamido)pyridin-4-yl)oxy)phenyl)-
-N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide (50 mg, 21%
yield). .sup.1H NMR (400 MHz, Methanol-d.sub.4): .delta.: 8.21-8.16
(m, 2H), 7.65 (d, J=2.4 Hz, 1H), 7.55-7.51 (m, 2H), 7.26 (dd,
J=10.4, 6.8 Hz, 1H), 7.10-7.05 (m, 2H), 6.76 (q, J=5.6, 2.4 Hz,
1H), 3.90 (q, J=6.8 Hz, 1H), 3.44, (s, 3H), 1.74-1.69 (m, 4H), 1.38
(d, J=6.8 Hz, 3H) [missing 3 NH]; MS (ESI): m/z 528.9
[M+H].sup.+.
Example 18
[0206] To a solution of (S)-2-Acetoxy-propionic acid (300 mg, 2.278
mmol) and N-methylmorpholine (341 mg, 3.41 mmol) in anhydrous
CH.sub.2Cl.sub.2 was added drop wise isobutyl chloroformate (457
mg, 3.41 mmol). The resultant mixture was stirred at 25.degree. C.
for 1 h.
N-(4-((2-aminopyridin-4-yl)oxy)-2,5-difluorophenyl)-N'-(4-fluorophenyl)cy-
clopropane-1,1-dicarboxamide (250 mg, 0.578 mmol) was added and the
resulting mixture was stirred at 25.degree. C. for 12 h. The
reaction mixture was concentrated under reduced pressure to give
(S)-14(4-(2,5-difluoro-4-(14(4-fluorophenyl)carbamoyl)cyclopropanecarboxa-
mido)phenoxy)pyridin-2-yl)amino)-1-oxopropan-2-yl acetate (180 mg,
55.9% yield) which was without further purification.
[0207] To a solution of
(S)-1-(4-(2,5-difluoro-4-(1-((4-fluorophenyl)carbamoyl)cyclopropanecarbox-
amido)phenoxy)pyridin-2-yl)amino)-1-oxopropan-2-yl acetate (180 mg,
0.323 mmol) in a solution of MeOH--H.sub.2O (3:1, 20 mL) was added
potassium carbonate (111.6 mg, 0.809 mmol), the mixture was stirred
at 25.degree. C. for 12 h. The reaction mixture was concentrated
and purified by prep-TLC to give
(S)--N-(2,5-difluoro-4-((2-(2-hydroxypropanamido)pyridin-4-yl)oxy)phenyl)-
-N'-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide (100 mg, 60%
yield). .sup.1H NMR (400 MHz, Methanol-d.sub.4): .delta. 8.21-8.16
(m, 2H), 7.78 (d, J=2.4 Hz, 1H), 7.54-7.51 (m, 2H), 7.28-7.24 (m,
1H), 7.10-7.05 (m, 2H), 6.76-6.74 (dd, J=6.0, 2.4 Hz, 1H), 4.23 (q,
J=6.8 Hz, 1H), 1.75-1.67 (m, 4H), 1.39 (d, J=6.8 Hz, 3H); MS (ESI):
m/z 515.2 [M+H].sup.+.
Example 19
[0208] To a solution of
N-(4-((2-aminopyridin-4-yl)oxy)-2,5-difluorophenyl)-N'-(4-fluorophenyl)cy-
clopropane-1,1-dicarboxamide (250 mg, 0.565 mmol) and
2-fluoro-2-methylpropanoic acid (108 mg, 1.02 mmol) in anhydrous
CH.sub.2Cl.sub.2 (30 mL) was added HATU (323 mg, 0.85 mmol) and
DIPEA (2.5 mL) under N.sub.2. This reaction mixture was stirred at
room temperature overnight. The solvent was removed in vacuo. The
residue was poured into water (100 mL), extracted with ethyl
acetate (3.times.50 mL), washed with brine, dried over anhydrous
sodium sulfate and concentrated in vacuo. This crude product was
purified by silica gel chromatography to give
N-(2,5-difluoro-4-((2-(2-fluoro-2-methylpropanamido)pyridin-4-yl)oxy-
)phenyl)-N'-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide (128 mg,
43% yield). .sup.1H-NMR (400 MHz, Methanol-d.sub.4): .delta.
8.21-8.17 (m, 2H), 7.73 (d, J=2.4 Hz, 1H), 7.54-7.51 (m, 2H), 7.23
(dd, J=10.8, 7.2 Hz, 1H), 7.08-7.04 (m, 2H), 6.75 (dd, J=5.6, 2.4
Hz, 1H), 1.76-1.69 (m, 4H), 1.62 (s, 3H), 1.56 (s, 3H); MS (ESI):
m/z 531.1 [M+H].sup.+.
Biological Data
[0209] c-MET Kinase Assay
[0210] Activity of e-MET kinase (Seq. ID No. 2) was determined by
following the production of ADP from the kinase reaction through
coupling with the pyruvate kinase/lactate dehydrogenase system
(e.g., Schindler et al. Science 2000, 289, pp. 1938-1942). In this
assay, the oxidation of NADH (thus the decrease at A340 nm) was
continuously monitored spectrophotometrically. The reaction mixture
(100 .mu.l) contained c-MET (c-MET residues: 956-1390, from
Invitrogen, catalogue #PV3143, 6 nM), polyE4Y (1 mg/mL), MgCl.sub.2
(10 mM), pyruvate kinase (4 units), lactate dehydrogenase (0.7
units), phosphoenol pyruvate (1 mM), and NADH (0.28 mM) in 90 mM
Tris buffer containing 0.25 mM DTT, 0.2% octyl-glucoside and 1%
DMSO, pH 7.5. Test compounds were incubated with c-MET (Seq. ID No.
2) and other reaction reagents at 22.degree. C. for 0.5 h before
ATP (100 .mu.M) was added to start the reaction. The absorption at
340 nm was monitored continuously for 2 hours at 30.degree. C. on
Polarstar Optima plate reader (BMG). The reaction rate was
calculated using the 1.0 to 2.0 h time frame. Percent inhibition
was obtained by comparison of reaction rate with that of a control
(i.e., with no test compound). IC.sub.50 values were calculated
from a series of percent inhibition values determined at a range of
inhibitor concentrations using software routines as implemented in
the GraphPad Prism software package.
TABLE-US-00001 c-MET Kinase (Seq ID No. 2)
MSYYHHHHHHDYDIPTTENLYFQGAMLVPRGSPWIPFTMKKRKQIKDLGSELVRYDARV
HTPHLDRLVSARSVSPTTEMVSNESVDYRATFPEDQFPNSSQNGSCRQVQYPLTDMSPILTS
GDSDISSPLLQNTVHIDLSALNPELVQAVQHVVIGPSSLIVHFNEVIGRGHFGCVYHGTLLD
NDGKKIHCAVKSLNRITDIGEVSQFLTEGIIMKDFSHPNVLSLLGICLRSEGSPLVVLPYMKH
GDLRNFIRNETHNPTVKDLIGFGLQVAKGMKYLASKKFVHRDLAARNCMLDEKFTVKVA
DFGLARDMYDKEYYSVHNKTGAKLPVKWMALESLQTQKFTTKSDVWSFGVLLWELMTR
GAPPYPDVNTFDITVYLLQGRRLLQPEYCPDPLYEVMLKCWHPKAEMRPSFSELVSRISAIF
STFIGEHYVHVNATYVNVKCVAPYPSLLSSEDNADDEVDTRPASFWETS.
c-KIT Kinase Assay
[0211] Activity of c-KIT kinase (Seq. ID No. 1) was determined by
following the production of ADP from the kinase reaction through
coupling with the pyruvate kinase/lactate dehydrogenase system
(e.g., Schindler et al. Science 2000, 289, pp. 1938-1942). In this
assay, the oxidation of NADH (thus the decrease at A340 nm) was
continuously monitored spectrophotometrically. The reaction mixture
(100 .mu.l) contained c-KIT (cKIT residues T544-V976, from
ProQinase, 5.4 nM), polyE4Y (1 mg/mL), MgCl.sub.2 (10 mM), pyruvate
kinase (4 units), lactate dehydrogenase (0.7 units), phosphoenol
pyruvate (1 mM), and NADH (0.28 mM) in 90 mM Tris buffer containing
0.2% octyl-glucoside and 1% DMSO, pH 7.5. Test compounds were
incubated with c-KIT (Seq. ID No. 1) and other reaction reagents at
22.degree. C. for less than 2 min before ATP (200 .mu.M) was added
to start the reaction. The absorption at 340 nm was monitored
continuously for 0.5 hours at 30.degree. C. on Polarstar Optima
plate reader (BMG). The reaction rate was calculated using the 0 to
0.5 h time frame. Percent inhibition was obtained by comparison of
reaction rate with that of a control (i.e., with no test compound).
IC.sub.50 values were calculated from a series of percent
inhibition values determined at a range of inhibitor concentrations
using software routines as implemented in the GraphPad Prism
software package.
TABLE-US-00002 c-KIT with N-terminal GST fusion (Seq ID No. 1)
LGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELGLEFPNLPYYIDGDVK
LTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVDIRYGVSRIAYSKDFETLKVDFLSKLP
EMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYMDPMCLDAFPKLVCFKKRIEAI
PQIDKYLKSSKYIWPLQGWQATFGGGDHPPKSDLVPRHNQTSLYKKAGSAAAVLEENLYF
QGTYKYLQKPMYEVQWKVVEEINGNNYVYIDPTQLPYDHKWEFPRNRLSFGKTLGAGAF
GKVVEATAYGLIKSDAAMTVAVKMLKPSAHLTEREALMSELKVLSYLGNHMNIVNLLGA
CTIGGPTLVITEYCCYGDLLNFLRRKRDSFICSKQEDHAEAALYKNLLHSKESSCSDSTNEY
MDMKPGVSYVVPTKADKRRSVRIGSYIERDVTPAIMEDDELALDLEDLLSFSYQVAKGMA
FLASKNCIHRDLAARNILLTHGRITKICDFGLARDIKNDSNYVVKGNARLPVKWMAPESIF
NCVYTFESDVWSYGIFLWELFSLGSSPYPGMPVDSKFYKMIKEGFRMLSPEHAPAEMYDI
MKTCWDADPLKRPTFKQIVQLIEKQISESTNHIYSNLANCSPNRQKPVVDHSVRINSVGST
ASSSQPLLVHDDV.
KDR Kinase Assay
Assay K1
[0212] The activity of KDR kinase was determined by following the
production of ADP from the kinase reaction through coupling with
the pyruvate kinase/lactate dehydrogenase system (e.g., Schindler
et al. Science 2000, 289, pp. 1938-1942). In this assay, the
oxidation of NADH (thus the decrease at A340 nm) was continuously
monitored spectrophotometrically. The reaction mixture (100 .mu.l)
contained KDR (Seq ID No. 3, 1.5 nM to 7.1 nM, nominal
concentration), polyE4Y (1 mg/mL), pyruvate kinase (3.5 units),
lactate dehydrogenase (5.5 units), phosphoenolpyruvate (1 mM), and
NADH (0.28 mM) in 60 mM Iris buffer containing 0.13%
octyl-glucoside, 13 mM MgCl.sub.2, 6.8 mM DTT, and 3.5% DMSO at pH
7.5. The reaction was initiated by adding ATP (0.2 mM, final
concentration). The absorption at 340 nm was continuously monitored
for 3 h at 30.degree. C. on a Polarstar Optima plate reader (BMG)
or instrument of similar capacity. The reaction rate was calculated
using the 1 h to 2 h time frame. Percent inhibition was obtained by
comparison of reaction rate with that of a control (i.e., with no
test compound). IC.sub.50 values were calculated from a series of
percent inhibition values determined at a range of inhibitor
concentrations using software routines as implemented in the
GraphPad Prism software package.
Assay K2
[0213] KDR kinase assay K2 is the same as for assay K1 except that
(1) a nominal concentration of 2.1 nM of enzyme was employed (2)
the reaction was pre-incubated at 30.degree. C. for 2 h prior to
initiation with ATP and (3) 1.0 mM ATP (final concentration) was
used to initiate the reaction.
Assay K3
[0214] KDR kinase assay K3 is the same as for assay K1 except that
(1) a nominal concentration of 1.1 nM of enzyme was employed, (2)
the buffer components per 100 .mu.l reaction mixture were as
follows: 75 mM Tris buffer containing 0.066% octyl-glucoside, 17 mM
MgCl.sub.2, and 1% DMSO at pH 7.5, (3) the final concentration of
DTT was 0.66 mM, (4) the reaction was pre-incubated at 30.degree.
C. for 1 h prior to initiation with ATP, and (5) 1.0 mM ATP (final
concentration) was used to initiate the reaction.
TABLE-US-00003 KDR protein sequence used for screening (Seq. ID No.
3) DPDELPLDEHCERLPYDASKWEFPRDRLKLGKPLGRGAFGQVIEADAFGIDKTATCRTVAVKML
KEGATHSEHRALMSELKILIHIGHHLNVVNLLGACTKPGGPLMVIVEFCKFGNLSTYLRSKRNEF
VPYKVAPEDLYKDFLTLEHLICYSFQVAKGMEFLASRKCIHRDLAARNILLSEKNVVKICDFGLA
RDIYKDPDYVRKGDARLPLKWMAPETIFDRVYTIQSDVWSFGVLLWEIFSLGASPYPGVKIDEEF
CRRLKEGTRMRAPDYTTPEMYQTMLDCWHGEPSQRPTFSELVEHLGNLLQANAQQD
FMS kinase Assay
[0215] Activity of FMS kinase was determined by following the
production of ADP from the kinase reaction through coupling with
the pyruvate kinase/lactate dehydrogenase system (e.g. Schindler et
al. Science 2000, 289, pp. 1938-1942). In this assay, the oxidation
of NADH (thus the decrease at A340 nm) was continuously monitored
spectrophotometrically. The reaction mixture (100 .mu.l) contained
FMS (purchased from Invitrogen or Millipore, 6 nM), polyE4Y (1
mg/mL), MgCl.sub.2 (10 mM), pyruvate kinase (4 units), lactate
dehydrogenase (0.7 units), phosphoenol pyruvate (1 mM) and NADH
(0.28 mM) and ATP (500 mM) in a 90 mM Tris buffer containing 0.2%
octyl-glucoside and 1% DMSO, pH 7.5. The inhibition reaction was
started by mixing serial diluted test compound with the above
reaction mixture. The absorption at 340 nm was monitored
continuously for 4 hours at 30.degree. C. on a Polarstar Optima or
Synergy 2 plate reader. The reaction rate was calculated using the
2 to 3 h time frame. Percent inhibition was obtained by comparison
of reaction rate with that of a control (i.e., with no test
compound). IC.sub.50 values were calculated from a series of
percent inhibition values determined at a range of inhibitor
concentrations using software routine's as implemented in the
GraphPad Prism software package.
EBC-1 Cell Culture
[0216] EBC-1 cells (catalog #JCRB0820) were obtained from the Japan
Health Science Research Resources Bank, Osaka, Japan. Briefly,
cells were grown in DMEM supplemented with 10% characterized fetal
bovine serum (Invitrogen, Carlsbad, Calif.) at 37.degree. C., 5%
CO.sub.2, 95% humidity. Cells were allowed to expand until reaching
70-95% confluency at which point they were subcultured or harvested
for assay use.
EBC-1 Cell Proliferation Assay
[0217] A serial dilution of test compound was dispensed into a
96-well black clear bottom plate (Corning, Corning, N.Y.). For each
cell line, five thousand cells were added per well in 200 .mu.L
complete growth medium. Plates were incubated for 67 hours at
37.degree. C., 5% CO.sub.2, 95% humidity. At the end of the
incubation period 40 .mu.L of a 440 .mu.M solution of resazurin
(Sigma, St. Louis, Mo.) in PBS was added to each well and incubated
for an additional 5 hours at 37.degree. C., 5% CO.sub.2, 95%
humidity. Plates were read on a Synergy2 reader (Biotek, Winooski,
Vt.) using an excitation of 540 nM and an emission of 600 nM. Data
was analyzed using Prism software (GraphPad, San Diego, Calif.) to
calculate IC.sub.50 values.
MKN-45 Cell Culture
[0218] MKN-45 cells (catalog #JCRB0254) were obtained from the
Japan Health Science Research Resources Bank, Osaka, Japan.
Briefly, cells were grown in RPMI 1640 media supplemented with 10%
characterized fetal bovine serum (Invitrogen, Carlsbad, Calif.) at
37.degree. C., 5% CO2, 95% humidity. Cells were allowed to expand
until reaching 70-95% confluency at which point they were
subcultured or harvested for assay use.
MKN-45 Cell Proliferation Assay
[0219] A serial dilution of test compound was dispensed into a
96-well black clear bottom plate (Corning, Corning, N.Y.). Five
thousand cells were added per well in 200 .mu.L complete growth
medium. Plates were incubated for 67 hours at 37.degree. C., 5%
CO.sub.2, 95% humidity. At the end of the incubation period 40
.mu.L of a 440 .mu.M solution of resazurin (Sigma, St. Louis, Mo.)
in PBS was added to each well and plates were incubated for an
additional 5 h at 37.degree. C., 5% CO.sub.2, 95% humidity. Plates
were read on a Synergy2 reader (Biotek, Winooski, Vt.) using an
excitation of 540 nM and an emission of 600 nM. Data was analyzed
using Prism software (GraphPad, San Diego, Calif.) to calculate
IC.sub.50 values.
RON Kinase Assay
[0220] Activity of RON kinase was determined by a radioactive
filtration binding assay where incorporation of .sup.33P from
.sup.33P-.gamma.-ATP to the substrate was measured. In this assay,
detection of .sup.33P was indicative of RON phosphorylation
activity which was directly proportional to the amount of
phosphorylated peptide substrate (KKSRGDYMTMQIG). Initially, the
reaction mixture contained: 400 nM RON, 20 mM HEPES, pH 7.5, 10 mM
MgCl.sub.2, 2 mM MnCl.sub.2, 1 mM EGTA, 0.02% Brij 35, 0.02 mg/ml .
. . BSA, 0.1 mM Na.sub.3VO.sub.4, and 2 mM DTT. The reaction
mixture was incubated with compound at room temperature for 30
minutes. To initiate the reaction, an equal volume of 20 .mu.M ATP
and 0.4 mg/mL peptide substrate were added and then incubated at
room temperature for 2 hours. The final assay conditions were: 200
nM RON, 10 .mu.M ATP, 0.2 mg/mL substrate 20 mM HEPES, pH 7.5, 10
mM MgCl.sub.2, 1 mM EGTA, 0.02% Brij 35, 0.02 mg/mL BSA, 0.1 mM
Na.sub.3VO.sub.4, 2 mM DTT, and 1% DMSO. IC.sub.50 values were
calculated from a series of % Activity values determined at a range
of inhibitor concentrations using software routines as implemented
in the GraphPad Prism software package.
RON Sequence/Protein Information (UniProtKB/Swiss-Prot Entry
Q04912)
[0221] GST tagged recombinant human RON, amino acids 983-1400.
Expressed in insect cells.
FLT1 Kinase Assay
[0222] Activity of FLT1 kinase was determined by a radioactive
filtration binding assay where incorporation of .sup.33P from
.sup.33P-.gamma.-ATP to the substrate is measured. In this assay,
detection of .sup.33P was indicative of FLT1 phosphorylation
activity which was directly proportional to the amount of
phosphorylated peptide substrate poly[Glu:Tyr] (4:1). Initially,
the reaction mixture contained: 400 nM FLT1, 20 mM HEPES, pH 7.5,
10 mM MgCl.sub.2, 2 mM MnCl.sub.2, 1 mM EGTA, 0.02% Brij 35, 0.02
mg mL BSA, 0.1 mM Na.sub.3VO.sub.4, and 2 mM DTT. The reaction
mixture was incubated with compound at room temperature for 30
minutes. To initiate the reaction, an equal volume of 20 .mu.M ATP
and 0.2 mg/mL peptide substrate were added and then incubated at
room temperature for 2 hours. The final assay conditions were: 200
nM FLT1, 10 .mu.M ATP, 0.1 mg/mL substrate 20 mM HEPES, pH 7.5, 10
mM MgCl.sub.2, 1 mM EGTA, 0.02% Brij 35, 0.02 mg/mL BSA, 0.1 mM
Na.sub.3VO.sub.4, 2 mM DTT, and 1% DMSO. IC.sub.50 values were
calculated from a series of % Activity values determined at a range
of inhibitor concentrations using software routines as implemented
in the Graph Pad Prism software package.
FLT1 Sequence/Protein Information (UniProtKB/Swiss-Prot Entry
P17948)
[0223] GST tagged recombinant human FLT1, amino acids 781-1338.
Expressed in insect cells.
RET Kinase Assay
[0224] Activity of RET kinase was determined by following the
production of ADP from the kinase reaction through coupling with
the pyruvate kinase/lactate dehydrogenase system (see e.g.,
Schindler et al. Science (2000) 289: 1938-1942). In this assay, the
oxidation of NADH (thus the decrease at A340 nm) was continuously
monitored spectrophotometrically. The reaction mixture (100 .mu.l)
contained RET (amino acid residues 658-1114, from Invitrogen, 2
nM), polyE4Y (1.5 mg/ml), MgCl.sub.2 (18 mM), pyruvate kinase (4
units), lactate dehydrogenase (0.7 units), phosphoenol pyruvate (1
mM), and NADH (0.28 mM) in 90 mM iris buffer containing 0.2%
octyl-glucoside, 1 mM DTT and 1% DMSO, pH 7.5. Test compounds were
incubated with RET kinase and other reaction reagents at 22.degree.
C. for <2 min before ATP (500 .mu.M) was added to start the
reaction. The absorption at 340 nm was monitored continuously for 3
hours at 30.degree. C. on BioTek Synergy 2 Reader. The reaction
rate was calculated using the 1 to 2 h time frame. Percent
inhibition was obtained by comparison of reaction rate with that of
a control (i.e., with no test compound). IC.sub.50 values were
calculated from a series of percent inhibition values determined at
a range of inhibitor concentrations using software routines as
implemented in the Graph Pad Prism software package.
RET Sequence/Protein Information (UniProtKB/Swiss-Prot Entry
P07949)
[0225] GST tagged recombinant human RET, amino acids 658-1114.
Expressed in insect cells.
[0226] Compounds, of Formula I exhibit inhibitory activity in one
or more of the aforementioned assays when evaluated at
concentrations .ltoreq.10 .mu.M.
[0227] An unexpected increase in potency and/or selectivity is
observed when the central phenyl ring of the compounds disclosed
herein contains a distinct para-di-substitution pattern. In
addition, it is theorized that the presence of certain R3 moieties
on the monocyclic nitrogen-containing heteroaromatic ring work in
concert with the para-di-substitution pattern of the central phenyl
ring to unexpectedly give rise to further improvement in potency
and/or selectivity. It is theorized that the identity and location
of certain moieties on the heteroaromatic ring relative to the
ether oxygen linker and a ring nitrogen atom contribute to these
results. For example, in the compounds described herein, the NH--R3
moiety is regiochemically located meta- to the ether oxygen linker
and ortho- to a ring nitrogen atom.
[0228] The unexpected potency and selectivity of compounds having
the characteristics of Formula I are exemplified in the data
presented in Table 1. Compound F, Compound G, and Compound H are
disclosed in U.S. Patent Publication No. 2008/0319188 A1
(hereinafter "the '188 Application"). The data for Compound F,
Compound G, and Compound H are taken from the values published in
the '188 application (pp. 96-97) and in U.S. Patent Publication No.
2009/0227556 A1 (hereinafter "the '556 application"). The data for
Compound D (Example 1) and Compound E (Example 8) were obtained by
the methods described in the Biological Data section, below.
TABLE-US-00004 TABLE 1 Fold Fold Fold Fold Selectivity Selectivity
MET RON Selectivity Selectivity VEGFR-1 VEGR-1/ VEGFR-2 VEGFR-2/
Inhibitor IC.sub.50 IC.sub.50 RON/MET RET IC.sub.50 RET/MET
IC.sub.50 MET IC.sub.50 MET Compound F 53 nM 17 nM 0.32 130 nM 2.45
88 nM 1.66 240 nM 4.53 Compound G 47 nM 2 nM 0.04 38 nM 0.81 21 nM
0.45 100 nM 2.13 Compound H 4 nM 3 nM 0.75 28 nM 7 14 nM 3.5 10 nM
2.5 Compound D 4 nM 5,000 nM >1,250 >3,300 nM >825 79 nM
19.75 52 nM 13 Compound E 26 nM 100 nM 3.85 43 nM 1.65 160 nM 6.15
122 nM 4.69
[0229] The structures of Compound F, Compound G, Compound H,
Compound D, and Compound E are below.
##STR00029##
[0230] Cyclopropane diamide Compound F, Compound G, and Compound H
were disclosed in the '188 Application as Examples 15, 92, and 91,
respectively. As shown in Table 1, Compound F and Compound G were
reported to inhibit MET kinase biochemical activity with comparable
IC.sub.50 values of 53 nM and 47 nM, respectively.
[0231] The structures of Compound F and Compound G are almost
identical, with the exception that Compound F is mono-fluorinated
in the central phenyl ring whereas Compound G is di-fluorinated
wherein the two fluorines are oriented para- with respect to each
other in the central phenyl ring.
[0232] In contrast, it has unexpectedly been found that Compound D,
disclosed herein, potently inhibits c-MET kinase with an IC.sub.50
value of 4 nM. Compound D is 6.5-fold more potent versus c-MET
kinase than its mono-fluoro analog Compound E (4 nM versus 26 nM;
see Table 1). This 6.5-fold greater potency versus MET kinase is
unexpected in view of the c-MET inhibition data for Compound G
compared to its corresponding mono-fluoro analog Compound F. As
reported in the '188 Application, Compound G and Compound F exhibit
essentially equivalent potency versus c-MET kinase (53 nM versus 47
nM, respectively; 1.1-fold ratio of potency). See the '188
application, pp. 96-97. Compound H is also reported to be a potent
MET kinase inhibitor, with IC.sub.50 of 4 nM. Id. Compound H, like
Compound D, is di-fluorinated, with the two fluorines being
oriented para with respect to each other in the central phenyl
ring. Compound H, however, does not exhibit the selectivity against
c-MET inhibition that has been observed for Compound D.
[0233] It has also been unexpectedly been found that Compound D
exhibits a much higher kinase selectivity versus RON, RET, VEGFR-1,
and VEGFR-2 kinases, compared to Compound E. See Table 1. RON is a
very close kinase of the MET subfamily of kinases, and inhibitors
of MET kinase are often not selective versus RON. Whereas Compound
D is >1,250 fold selective of MET kinase versus RON kinase,
Compound E is only 3.85 fold of MET kinase versus RON kinase. See
Table 1. Also, whereas Compound D is >825 fold selective of MET
kinase versus RET kinase, Compound E is only 1.65 fold selective of
MET kinase versus RET kinase. Additionally, whereas Compound D is
19.75-fold selective of MET kinase versus VEGFR-1 kinase, Compound
E is significantly less selective: 6.15-fold selective of MET
kinase versus VEGFR-1 kinase. Finally, whereas Compound D is
13-fold selective of MET kinase versus VEGFR-2 kinase, Compound E
is significantly less selective: 4.69-fold selective of MET kinase
versus VEGFR-2 kinase.
[0234] In contrast, as illustrated in Table 1, Compound F, Compound
G, and Compound H do not exhibit selectivity of c-MET versus RON
kinase, as evidenced by reported IC.sub.50 values versus RON of 17
nM, 2 nM, and 3 nM, respectively. See Table 1, below and the '556
application, pg. 36. Moreover, as shown in Table 1, the Fold
Selectivity of RON relative to MET kinase is 0.32, 0.04, and 0.75,
respectively, for Compound F, Compound G, and Compound H,
indicating that the compounds are more potent against RON kinase
than against MET kinase. In view of these data, the selectivity of
Compound D versus these `off target` kinases is unexpected. In
summary, it is theorized that the presence of the central phenyl
ring para-di-substitution pattern, in combination with the pyridine
ring acetamide moiety, confers this unexpected MET potency and
selectivity versus these `off targets.`
[0235] Without wishing to be bound by a particular theory, it is
theorized that the presence of certain non-hydrogen R3 moieties on
the monocyclic nitrogen-containing heteroaromatic ring work in
concert with the para-di-substitution pattern of the central phenyl
ring to unexpectedly give rise to further improvement in potency
and/or selectivity. Specifically, one difference in the structure
of Compound D, Compound G, and Compound H resides in the identity
of the substituent in the pyridine ring. In Compound D, this
substituent is --NHC(O)CH3 (an acetamide), whereas in Compound G
and Compound H the substituent is a more extended urea. In summary,
it is theorized that the combination of the presence of the central
phenyl ring para-di-substitution pattern, in combination with the
specific pyridine ring moiety (e.g., acetamide vs. extended urea
moieties), confers this unexpected c-MET potency and high
selectivity versus these `off targets.` Other features which may
also contribute to the unexpected results described herein include:
a para-regiochemical relationship between the oxygen ether linker
and the nitrogen amide atoms attached to the central phenyl ring; a
regiochemical para-relationship between the oxygen ether linker
atom and the ring nitrogen in the nitrogen-containing ring; and
absence of an alkyl spacer between the aromatic ring and the
nitrogen of the cyclopropane carbonyl. While cyclopropane amides
have been reported in the literature as inhibitors of c-MET kinase
activity, compounds having the features discussed above have not
been disclosed.
EQUIVALENTS
[0236] Those skilled in the art will recognize, or be able to
ascertain, using no more than routine experimentation, numerous
equivalents to the specific embodiments described specifically in
this disclosure. Such equivalents are intended to be encompassed in
the scope of the following claims.
Sequence CWU 1
1
41676PRTArtificial Sequencec-KIT with N-terminal GST fusion 1Leu
Gly Tyr Trp Lys Ile Lys Gly Leu Val Gln Pro Thr Arg Leu Leu1 5 10
15Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu Arg Asp
20 25 30Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu Gly Leu Glu
Phe 35 40 45Pro Asn Leu Pro Tyr Tyr Ile Asp Gly Asp Val Lys Leu Thr
Gln Ser 50 55 60Met Ala Ile Ile Arg Tyr Ile Ala Asp Lys His Asn Met
Leu Gly Gly65 70 75 80Cys Pro Lys Glu Arg Ala Glu Ile Ser Met Leu
Glu Gly Ala Val Asp 85 90 95Ile Arg Tyr Gly Val Ser Arg Ile Ala Tyr
Ser Lys Asp Phe Glu Thr 100 105 110Leu Lys Val Asp Phe Leu Ser Lys
Leu Pro Glu Met Leu Lys Met Phe 115 120 125Glu Asp Arg Leu Cys His
Lys Thr Tyr Leu Asn Gly Asp His Val Thr 130 135 140His Pro Asp Phe
Met Leu Tyr Asp Ala Leu Asp Val Val Leu Tyr Met145 150 155 160Asp
Pro Met Cys Leu Asp Ala Phe Pro Lys Leu Val Cys Phe Lys Lys 165 170
175Arg Ile Glu Ala Ile Pro Gln Ile Asp Lys Tyr Leu Lys Ser Ser Lys
180 185 190Tyr Ile Trp Pro Leu Gln Gly Trp Gln Ala Thr Phe Gly Gly
Gly Asp 195 200 205His Pro Pro Lys Ser Asp Leu Val Pro Arg His Asn
Gln Thr Ser Leu 210 215 220Tyr Lys Lys Ala Gly Ser Ala Ala Ala Val
Leu Glu Glu Asn Leu Tyr225 230 235 240Phe Gln Gly Thr Tyr Lys Tyr
Leu Gln Lys Pro Met Tyr Glu Val Gln 245 250 255Trp Lys Val Val Glu
Glu Ile Asn Gly Asn Asn Tyr Val Tyr Ile Asp 260 265 270Pro Thr Gln
Leu Pro Tyr Asp His Lys Trp Glu Phe Pro Arg Asn Arg 275 280 285Leu
Ser Phe Gly Lys Thr Leu Gly Ala Gly Ala Phe Gly Lys Val Val 290 295
300Glu Ala Thr Ala Tyr Gly Leu Ile Lys Ser Asp Ala Ala Met Thr
Val305 310 315 320Ala Val Lys Met Leu Lys Pro Ser Ala His Leu Thr
Glu Arg Glu Ala 325 330 335Leu Met Ser Glu Leu Lys Val Leu Ser Tyr
Leu Gly Asn His Met Asn 340 345 350Ile Val Asn Leu Leu Gly Ala Cys
Thr Ile Gly Gly Pro Thr Leu Val 355 360 365Ile Thr Glu Tyr Cys Cys
Tyr Gly Asp Leu Leu Asn Phe Leu Arg Arg 370 375 380Lys Arg Asp Ser
Phe Ile Cys Ser Lys Gln Glu Asp His Ala Glu Ala385 390 395 400Ala
Leu Tyr Lys Asn Leu Leu His Ser Lys Glu Ser Ser Cys Ser Asp 405 410
415Ser Thr Asn Glu Tyr Met Asp Met Lys Pro Gly Val Ser Tyr Val Val
420 425 430Pro Thr Lys Ala Asp Lys Arg Arg Ser Val Arg Ile Gly Ser
Tyr Ile 435 440 445Glu Arg Asp Val Thr Pro Ala Ile Met Glu Asp Asp
Glu Leu Ala Leu 450 455 460Asp Leu Glu Asp Leu Leu Ser Phe Ser Tyr
Gln Val Ala Lys Gly Met465 470 475 480Ala Phe Leu Ala Ser Lys Asn
Cys Ile His Arg Asp Leu Ala Ala Arg 485 490 495Asn Ile Leu Leu Thr
His Gly Arg Ile Thr Lys Ile Cys Asp Phe Gly 500 505 510Leu Ala Arg
Asp Ile Lys Asn Asp Ser Asn Tyr Val Val Lys Gly Asn 515 520 525Ala
Arg Leu Pro Val Lys Trp Met Ala Pro Glu Ser Ile Phe Asn Cys 530 535
540Val Tyr Thr Phe Glu Ser Asp Val Trp Ser Tyr Gly Ile Phe Leu
Trp545 550 555 560Glu Leu Phe Ser Leu Gly Ser Ser Pro Tyr Pro Gly
Met Pro Val Asp 565 570 575Ser Lys Phe Tyr Lys Met Ile Lys Glu Gly
Phe Arg Met Leu Ser Pro 580 585 590Glu His Ala Pro Ala Glu Met Tyr
Asp Ile Met Lys Thr Cys Trp Asp 595 600 605Ala Asp Pro Leu Lys Arg
Pro Thr Phe Lys Gln Ile Val Gln Leu Ile 610 615 620Glu Lys Gln Ile
Ser Glu Ser Thr Asn His Ile Tyr Ser Asn Leu Ala625 630 635 640Asn
Cys Ser Pro Asn Arg Gln Lys Pro Val Val Asp His Ser Val Arg 645 650
655Ile Asn Ser Val Gly Ser Thr Ala Ser Ser Ser Gln Pro Leu Leu Val
660 665 670His Asp Asp Val 6752474PRTHomo sapiens 2Met Ser Tyr Tyr
His His His His His His Asp Tyr Asp Ile Pro Thr1 5 10 15Thr Glu Asn
Leu Tyr Phe Gln Gly Ala Met Leu Val Pro Arg Gly Ser 20 25 30Pro Trp
Ile Pro Phe Thr Met Lys Lys Arg Lys Gln Ile Lys Asp Leu 35 40 45Gly
Ser Glu Leu Val Arg Tyr Asp Ala Arg Val His Thr Pro His Leu 50 55
60Asp Arg Leu Val Ser Ala Arg Ser Val Ser Pro Thr Thr Glu Met Val65
70 75 80Ser Asn Glu Ser Val Asp Tyr Arg Ala Thr Phe Pro Glu Asp Gln
Phe 85 90 95Pro Asn Ser Ser Gln Asn Gly Ser Cys Arg Gln Val Gln Tyr
Pro Leu 100 105 110Thr Asp Met Ser Pro Ile Leu Thr Ser Gly Asp Ser
Asp Ile Ser Ser 115 120 125Pro Leu Leu Gln Asn Thr Val His Ile Asp
Leu Ser Ala Leu Asn Pro 130 135 140Glu Leu Val Gln Ala Val Gln His
Val Val Ile Gly Pro Ser Ser Leu145 150 155 160Ile Val His Phe Asn
Glu Val Ile Gly Arg Gly His Phe Gly Cys Val 165 170 175Tyr His Gly
Thr Leu Leu Asp Asn Asp Gly Lys Lys Ile His Cys Ala 180 185 190Val
Lys Ser Leu Asn Arg Ile Thr Asp Ile Gly Glu Val Ser Gln Phe 195 200
205Leu Thr Glu Gly Ile Ile Met Lys Asp Phe Ser His Pro Asn Val Leu
210 215 220Ser Leu Leu Gly Ile Cys Leu Arg Ser Glu Gly Ser Pro Leu
Val Val225 230 235 240Leu Pro Tyr Met Lys His Gly Asp Leu Arg Asn
Phe Ile Arg Asn Glu 245 250 255Thr His Asn Pro Thr Val Lys Asp Leu
Ile Gly Phe Gly Leu Gln Val 260 265 270Ala Lys Gly Met Lys Tyr Leu
Ala Ser Lys Lys Phe Val His Arg Asp 275 280 285Leu Ala Ala Arg Asn
Cys Met Leu Asp Glu Lys Phe Thr Val Lys Val 290 295 300Ala Asp Phe
Gly Leu Ala Arg Asp Met Tyr Asp Lys Glu Tyr Tyr Ser305 310 315
320Val His Asn Lys Thr Gly Ala Lys Leu Pro Val Lys Trp Met Ala Leu
325 330 335Glu Ser Leu Gln Thr Gln Lys Phe Thr Thr Lys Ser Asp Val
Trp Ser 340 345 350Phe Gly Val Leu Leu Trp Glu Leu Met Thr Arg Gly
Ala Pro Pro Tyr 355 360 365Pro Asp Val Asn Thr Phe Asp Ile Thr Val
Tyr Leu Leu Gln Gly Arg 370 375 380Arg Leu Leu Gln Pro Glu Tyr Cys
Pro Asp Pro Leu Tyr Glu Val Met385 390 395 400Leu Lys Cys Trp His
Pro Lys Ala Glu Met Arg Pro Ser Phe Ser Glu 405 410 415Leu Val Ser
Arg Ile Ser Ala Ile Phe Ser Thr Phe Ile Gly Glu His 420 425 430Tyr
Val His Val Asn Ala Thr Tyr Val Asn Val Lys Cys Val Ala Pro 435 440
445Tyr Pro Ser Leu Leu Ser Ser Glu Asp Asn Ala Asp Asp Glu Val Asp
450 455 460Thr Arg Pro Ala Ser Phe Trp Glu Thr Ser465
4703315PRTHomo sapiens 3Asp Pro Asp Glu Leu Pro Leu Asp Glu His Cys
Glu Arg Leu Pro Tyr1 5 10 15Asp Ala Ser Lys Trp Glu Phe Pro Arg Asp
Arg Leu Lys Leu Gly Lys 20 25 30Pro Leu Gly Arg Gly Ala Phe Gly Gln
Val Ile Glu Ala Asp Ala Phe 35 40 45Gly Ile Asp Lys Thr Ala Thr Cys
Arg Thr Val Ala Val Lys Met Leu 50 55 60Lys Glu Gly Ala Thr His Ser
Glu His Arg Ala Leu Met Ser Glu Leu65 70 75 80Lys Ile Leu Ile His
Ile Gly His His Leu Asn Val Val Asn Leu Leu 85 90 95Gly Ala Cys Thr
Lys Pro Gly Gly Pro Leu Met Val Ile Val Glu Phe 100 105 110Cys Lys
Phe Gly Asn Leu Ser Thr Tyr Leu Arg Ser Lys Arg Asn Glu 115 120
125Phe Val Pro Tyr Lys Val Ala Pro Glu Asp Leu Tyr Lys Asp Phe Leu
130 135 140Thr Leu Glu His Leu Ile Cys Tyr Ser Phe Gln Val Ala Lys
Gly Met145 150 155 160Glu Phe Leu Ala Ser Arg Lys Cys Ile His Arg
Asp Leu Ala Ala Arg 165 170 175Asn Ile Leu Leu Ser Glu Lys Asn Val
Val Lys Ile Cys Asp Phe Gly 180 185 190Leu Ala Arg Asp Ile Tyr Lys
Asp Pro Asp Tyr Val Arg Lys Gly Asp 195 200 205Ala Arg Leu Pro Leu
Lys Trp Met Ala Pro Glu Thr Ile Phe Asp Arg 210 215 220Val Tyr Thr
Ile Gln Ser Asp Val Trp Ser Phe Gly Val Leu Leu Trp225 230 235
240Glu Ile Phe Ser Leu Gly Ala Ser Pro Tyr Pro Gly Val Lys Ile Asp
245 250 255Glu Glu Phe Cys Arg Arg Leu Lys Glu Gly Thr Arg Met Arg
Ala Pro 260 265 270Asp Tyr Thr Thr Pro Glu Met Tyr Gln Thr Met Leu
Asp Cys Trp His 275 280 285Gly Glu Pro Ser Gln Arg Pro Thr Phe Ser
Glu Leu Val Glu His Leu 290 295 300Gly Asn Leu Leu Gln Ala Asn Ala
Gln Gln Asp305 310 315413PRTUnknownRON kinase peptide substrate
4Lys Lys Ser Arg Gly Asp Tyr Met Thr Met Gln Ile Gly1 5 10
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