U.S. patent number RE46,097 [Application Number 14/544,396] was granted by the patent office on 2016-08-09 for pyrrole inhibitors of erk protein kinase, synthesis thereof and intermediates thereto.
This patent grant is currently assigned to Vertex Pharmaceuticals Incorporated. The grantee listed for this patent is Vertex Pharmaceuticals Incorporated. Invention is credited to Gabriel Martinez Botella, Michael Hale, Francois Maltais, Judith Straub, Qing Tang.
United States Patent |
RE46,097 |
Botella , et al. |
August 9, 2016 |
Pyrrole inhibitors of ERK protein kinase, synthesis thereof and
intermediates thereto
Abstract
The present invention relates to compounds useful of inhibitors
of protein kinases. The invention also provides pharmaceutically
acceptable compositions comprising said compounds and methods of
using the compositions in the treatment of various disease,
conditions, or disorders.
Inventors: |
Botella; Gabriel Martinez
(Wayland, MA), Hale; Michael (Bedford, MA), Maltais;
Francois (Tewksbury, MA), Straub; Judith (Ardmore,
PA), Tang; Qing (Acton, MA) |
Applicant: |
Name |
City |
State |
Country |
Type |
Vertex Pharmaceuticals Incorporated |
Boston |
MA |
US |
|
|
Assignee: |
Vertex Pharmaceuticals
Incorporated (Boston, MA)
|
Family
ID: |
35106760 |
Appl.
No.: |
14/544,396 |
Filed: |
August 23, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
|
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60571309 |
May 14, 2004 |
|
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Reissue of: |
11128870 |
May 13, 2005 |
7354939 |
Apr 8, 2008 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P
37/00 (20180101); A61P 29/00 (20180101); A61P
43/00 (20180101); A61P 15/00 (20180101); A61P
1/18 (20180101); A61P 9/10 (20180101); A61P
1/16 (20180101); A61P 19/08 (20180101); C07F
9/65583 (20130101); A61P 37/06 (20180101); C07D
401/04 (20130101); A61P 25/00 (20180101); A61P
1/04 (20180101); A61P 37/04 (20180101); A61P
17/00 (20180101); A61P 31/12 (20180101); A61P
37/02 (20180101); A61P 35/02 (20180101); A61P
9/00 (20180101); A61P 35/00 (20180101) |
Current International
Class: |
A61K
31/4439 (20060101); C07D 401/04 (20060101); C07F
9/6558 (20060101) |
Field of
Search: |
;514/343 ;546/279.1 |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
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EP |
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02064586 |
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Aug 2002 |
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WO |
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03091246 |
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Nov 2003 |
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WO |
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2004005283 |
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Jan 2004 |
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WO |
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2004009562 |
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Jan 2004 |
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WO |
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2004013125 |
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Feb 2004 |
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WO |
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2004016597 |
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Feb 2004 |
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WO |
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2004083203 |
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Sep 2004 |
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WO |
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Other References
Blanchard et al. "Hetarynic Synthesis and Chemical Transformation
of Dihydrodipyridopyrazines," Tetrahedron 58 (18): 3513-3524
(2002). cited by applicant .
PCT/US2005/016902 International Search Report. cited by
applicant.
|
Primary Examiner: Huang; Evelyn
Attorney, Agent or Firm: Finnegan, Henderson, Farabow,
Garrett & Dunner LLP
Parent Case Text
CROSS REFERENCE TO RELATED APPLICATIONS
This application claims priority to U.S. Provisional Patent
Application 60/571,309 filed May 14, 2004, the entire contents of
which are hereby incorporated herein by reference.
Claims
We claim:
.[.1. A compound of formula I: ##STR00049## or a pharmaceutically
acceptable salt thereof, wherein: R.sup.1 is a C.sub.1-6 aliphatic
group, wherein R.sup.1 is optionally substituted with up to 2
groups independently selected from --OR or --C.sub.1-3 haloalkyl;
each R is independently hydrogen or C.sub.1-4 aliphatic; R.sup.2 is
R, fluoro, or chloro; m is 0, 1, or 2; and R.sup.3 is hydrogen,
C.sub.1-3 aliphatic, fluoro, or chloro; wherein each aliphatic
group is, independently, a saturated or unsaturated straight or
branched hydrocarbon chain or monocyclic non-aromatic
hydrocarbon..].
.[.2. The compound according to claim 1, wherein R.sup.1 is
C.sub.1-4 aliphatic optionally substituted with --OR or --C.sub.1-3
haloalkyl..].
.[.3. The compound according to claim 2, wherein R.sup.1 is
C.sub.1-4 aliphatic optionally substituted with --OH, --CHF.sub.2,
--CH.sub.2F, or --CF.sub.3..].
.[.4. The compound according to claim 3, wherein R.sup.1 is
isopropyl, 2-butyl, cyclopropyl, or ethyl, wherein each moiety is
optionally substituted with --OH or --CF.sub.3..].
.[.5. The compound according to claim 1, wherein R.sup.2 is
hydrogen, C.sub.1-3 aliphatic, or chloro..].
.[.6. The compound according to claim 1, wherein R.sup.3 is
hydrogen, methyl, or chloro..].
.[.7. A compound selected from the group consisting of:
##STR00050## ##STR00051## ##STR00052## ##STR00053## .].
.[.8. A composition comprising a compound according to claim 1 and
a pharmaceutically acceptable carrier, adjuvant, or vehicle..].
.[.9. The compound according to claim 1, wherein: R.sup.1 is
isopropyl or 2-butyl, wherein R.sup.1 is optionally substituted
with one --OH; R.sup.2 is H or Cl; m is 1; and R.sup.3 is Cl or
methyl..].
.Iadd.10. The compound ##STR00054## .Iaddend.
.Iadd.11. A composition comprising a compound according to claim 10
and a pharmaceutically acceptable carrier, adjuvant, or
vehicle..Iaddend.
.Iadd.12. A pharmaceutically acceptable salt of the Compound
##STR00055## .Iaddend.
.Iadd.13. A composition comprising a pharmaceutically acceptable
salt of compound 1-9 according to claim 12 and a pharmaceutically
acceptable carrier, adjuvant, or vehicle..Iaddend.
Description
TECHNICAL FIELD OF INVENTION
The present invention relates to compounds useful as inhibitors of
protein kinases. The invention also provides pharmaceutically
acceptable compositions comprising the compounds of the invention
and methods of using the compositions in the treatment of various
disorders.
BACKGROUND OF THE INVENTION
The search for new therapeutic agents has been greatly aided in
recent years by a better understanding of the structure of enzymes
and other biomolecules associated with target diseases. One
important class of enzymes that has been the subject of extensive
study is protein kinases.
Protein kinases constitute a large family of structurally related
enzymes that are responsible for the control of a variety of signal
transduction processes within the cell. (See, Hardie, G. and Hanks,
S. (1995) The Protein Kinase Facts Book, I and II, Academic Press,
San Diego, Calif.). Protein kinases are thought to have evolved
from a common ancestral gene due to the conservation of their
structure and catalytic function. Almost all kinases contain a
similar 250-300 amino acid catalytic domain. The kinases may be
categorized into families by the substrates they phosphorylate
(e.g., protein-tyrosine, protein-serine/threonine, lipids, etc.).
Sequence motifs have been identified that generally correspond to
each of these kinase families (See, for example, Hanks, S. K.,
Hunter, T., FASEB J., 9:576-596 (1995); Knighton et al., Science,
253:407-414 (1991); Hiles et al., Cell, 70:419-429 (1992); Kunz et
al., Cell, 73:585-596 (1993); Garcia-Bustos et al., EMBO J.,
13:2352-2361 (1994)).
In general, protein kinases mediate intracellular signaling by
effecting a phosphoryl transfer from a nucleoside triphosphate to a
protein acceptor that is involved in a signaling pathway. These
phosphorylation events act as molecular on/off switches that can
modulate or regulate the target protein biological function. These
phosphorylation events are ultimately triggered in response to a
variety of extracellular and other stimuli. Examples of such
stimuli include environmental and chemical stress signals (e.g.,
osmotic shock, heat shock, ultraviolet radiation, bacterial
endotoxin, and H.sub.2O.sub.2), cytokines (e.g., interleukin-1
(IL-1) and tumor necrosis factor a (TNF-a)), and growth factors
(e.g., granulocyte macrophage-colony-stimulating factor (GM-CSF),
and fibroblast growth factor (FGF)). An extracellular stimulus may
affect one or more cellular responses related to cell growth,
migration, differentiation, secretion of hormones, activation of
transcription factors, muscle contraction, glucose metabolism,
control of protein synthesis, and regulation of the cell cycle.
Many diseases are associated with abnormal cellular responses
triggered by protein kinase-mediated events. These diseases include
autoimmune diseases, inflammatory diseases, bone diseases,
metabolic diseases, neurological and neurodegenerative diseases,
cancer, cardiovascular diseases, allergies and asthma, Alzheimer's
disease and hormone-related diseases. Accordingly, there has been a
substantial effort in medicinal chemistry to find protein kinase
inhibitors that are effective as therapeutic agents. However,
considering the lack of currently available treatment options for
the majority of the conditions associated with protein kinases,
there is still a great need for new therapeutic agents that inhibit
these protein targets.
Mammalian cells respond to extracellular stimuli by activating
signaling cascades that are mediated by members of the
mitogen-activated protein (MAP) kinase family, which include the
extracellular signal regulated kinases (ERKs), the p38 MAP kinases
and the c-Jun N-terminal kinases (JNKs). MAP kinases (MAPKs) are
activated by a variety of signals including growth factors,
cytokines, UV radiation, and stress-inducing agents. MAPKs are
serine/threonine kinases and their activation occur by dual
phosphorylation of threonine and tyrosine at the Thr-X-Tyr segment
in the activation loop. MAPKs phosphorylate various substrates
including transcription factors, which in turn regulate the
expression of specific sets of genes and thus mediate a specific
response to the stimulus.
ERK2 is a widely distributed protein kinase that achieves maximum
activity when both Thr183 and Tyr185 are phosphorylated by the
upstream MAP kinase kinase, MEK1 (Anderson et al., 1990, Nature
343, 651; Crews et al., 1992, Science 258, 478). Upon activation,
ERK2 phosphorylates many regulatory proteins, including the protein
kinases Rsk90 (Bjorbaek et al., 1995, J. Biol. Chem. 270, 18848)
and MAPKAP2 (Rouse et al., 1994, Cell 78, 1027), and transcription
factors such as ATF2 (Raingeaud et al., 1996, Mol. Cell Biol. 16,
1247), Elk-1 (Raingeaud et al. 1996), c-Fos (Chen et al., 1993
Proc. Natl. Acad. Sci. USA 90, 10952), and c-Myc (Oliver et al.,
1995, Proc. Soc. Exp. Biol. Med. 210, 162). ERK2 is also a
downstream target of the Ras/Raf dependent pathways (Moodie et al.,
1993, Science 260, 1658) and relays the signals from these
potentially oncogenic proteins. ERK2 has been shown to play a role
in the negative growth control of breast cancer cells (Frey and
Mulder, 1997, Cancer Res. 57, 628) and hyperexpression of ERK2 in
human breast cancer has been reported (Sivaraman et al., 1997, J.
Clin. Invest. 99, 1478). Activated ERK2 has also been implicated in
the proliferation of endothelin-stimulated airway smooth muscle
cells, suggesting a role for this kinase in asthma (Whelchel et
al., 1997, Am. J. Respir. Cell Mol. Biol. 16, 589).
Overexpression of receptor tyrosine kinases such as EGFR and ErbB2
(Arteaga C L, 2002, Semin Oncol. 29, 3-9; Eccles S A, 2001, J
Mammary Gland Biol Neoplasia 6:393-406; Mendelsohn J & Baselga
J, 2000, Oncogene 19, 6550-65), as well as activating mutations in
the Ras GTPase proteins (Nonage M & Siu L L, 2002, Curr Pharm
Des 8, 2231-42; Adjei A A, 2001, J Natl Cancer Inst 93, 1062-74) or
B-Raf mutants (Davies H. et al., 2002, Nature 417, 949-54; Brose et
al., 2002, Cancer Res 62, 6997-7000) are major contributors to
human cancer. These genetic alterations are correlated with poor
clinical prognosis and result in activation of the
Raf-1/2/3-MEK1/2-ERK1/2 signal transduction cascade in a broad
panel of human tumors. Activated ERK (i.e. ERK1 and/or ERK2) is a
central signaling molecule that has been associated with the
control of proliferation, differentiation, anchorage-independent
cell survival, and angiogenesis, contributing to a number of
processes that are important for the formation and progression of
malignant tumors. These data suggest that an ERK1/2 inhibitor will
exert pleiotropic activity, including proapoptotic,
anti-proliferative, anti-metastatic and anti-angiogenic effects,
and offer a therapeutic opportunity against a very broad panel of
human tumors.
There is a growing body of evidence that implicates constitutive
activation of the ERK MAPK pathway in the oncogenic behavior of
select cancers. Activating mutations of Ras are found in .about.30%
of all cancers, with some, such as pancreatic (90%) and colon (50%)
cancer, harboring particularly high mutation rates (ref). Ras
mutations have also been identified in 9-15% of melanomas, but
B-Raf somatic missense mutations conferring constitutive activation
are more frequent and found in 60-66% malignant melanomas.
Activating mutations of Ras, Raf and MEK are able to oncogenically
transform fibroblasts in vitro, and Ras or Raf mutations in
conjunction with the loss of a tumor suppressor gene (e.g.
p16INK4A) can cause spontaneous tumor development in vivo.
Increased ERK activity has been demonstrated in these models and
has also been widely reported in appropriate human tumors. In
melanoma, high basal ERK activity resulting from either B-Raf or
N-Ras mutations or autocrine growth factor activation is well
documented and has been associated with rapid tumor growth,
increased cell survival and resistance to apoptosis. Additionally,
ERK activation is considered a major driving force behind the
highly metastatic behavior of melanoma associated with increased
expression of both extracellular matrix degrading proteases and
invasion-promoting integrins as well as the downregulation of
E-cadherin adhesion molecules that normally mediate keratinocyte
interactions to control melanocyte growth. These data taken
together, indicate ERK as promising therapeutic target for the
treatment of melanoma, a currently untreatable disease.
SUMMARY OF THE INVENTION
It has now been found that compounds of this invention, and
pharmaceutically acceptable compositions thereof, are effective as
inhibitors of ERK protein kinase. These compounds have the general
formula I:
##STR00001## or a pharmaceutically acceptable salt thereof, wherein
m, R.sup.1, R.sup.2, and R.sup.3 are as defined below. These
compounds, and pharmaceutically acceptable compositions thereof,
are useful for treating or lessening the severity of a variety of
disorders, especially proliferative disorders such as cancer.
The compounds provided by this invention are also useful for the
study of kinases in biological and pathological phenomena and the
study of intracellular signal transduction pathways mediated by
such kinases, and the comparative evaluation of new kinase
inhibitors.
DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS OF THE INVENTION
1. General Description of Compounds of the Invention:
The present invention relates to a compound of formula I:
##STR00002## or a pharmaceutically acceptable salt thereof,
wherein: R.sup.1 is a C.sub.1-6 aliphatic group, wherein R.sup.1 is
optionally substituted with up to 2 groups independently selected
from --OR or --C.sub.1-3 haloalkyl; each R is independently
hydrogen or C.sub.1-4 aliphatic; each R.sup.2 is independently R,
fluoro, or chloro; m is 0, 1, or 2; and R.sup.3 is hydrogen,
C.sub.1-3 aliphatic, fluoro, or chloro. 2. Compounds and
Definitions:
Compounds of this invention include those described generally
above, and are further illustrated by the classes, subclasses, and
species disclosed herein. As used herein, the following definitions
shall apply unless otherwise indicated. For purposes of this
invention, the chemical elements are identified in accordance with
the Periodic Table of the Elements, CAS version, Handbook of
Chemistry and Physics, 75.sup.th Ed. Additionally, general
principles of organic chemistry are described in "Organic
Chemistry", Thomas Sorrell, University Science Books, Sausalito:
1999, and "March's Advanced Organic Chemistry", 5.sup.th Ed., Ed.:
Smith, M. B. and March, J., John Wiley & Sons, New York: 2001,
the entire contents of which are hereby incorporated by
reference.
As used herein, the term "prodrug" refers to a derivative of a
parent drug molecule that requires transformation within the body
in order to release the active drug, and that has improved physical
and/or delivery properties over the parent drug molecule. Prodrugs
are designed to enhance pharmaceutically and/or pharmacokinetically
based properties associated with the parent drug molecule. The
advantage of a prodrug lies in its physical properties, such as
enhanced water solubility for parenteral administration at
physiological pH compared to the parent drug, or it enhances
absorption from the digestive tract, or it may enhance drug
stability for long-term storage. In recent years several types of
bioreversible derivatives have been exploited for utilization in
designing prodrugs. Using esters as a prodrug type for drugs
containing carboxyl or hydroxyl function is known in the art as
described, for example, in "The Organic Chemistry of Drug Design
and Drug Interaction" Richard Silverman, published by Academic
Press (1992).
As described herein, compounds of the invention may optionally be
substituted with one or more substituents, such as are illustrated
generally above, or as exemplified by particular classes,
subclasses, and species of the invention. It will be appreciated
that the phrase "optionally substituted" is used interchangeably
with the phrase "substituted or unsubstituted." In general, the
term "substituted", whether preceded by the term "optionally" or
not, refers to the replacement of hydrogen radicals in a given
structure with the radical of a specified substituent. Unless
otherwise indicated, an optionally substituted group may have a
substituent at each substitutable position of the group, and when
more than one position in any given structure may be substituted
with more than one substituent selected from a specified group, the
substituent may be either the same or different at every
position.
Combinations of substituents envisioned by this invention are
preferably those that result in the formation of stable or
chemically feasible compounds. The term "stable", as used herein,
refers to compounds that are not substantially altered when
subjected to conditions to allow for their production, detection,
and preferably their recovery, purification, and use for one or
more of the purposes disclosed herein. In some embodiments, a
stable compound or chemically feasible compound is one that is not
substantially altered when kept at a temperature of 40.degree. C.
or less, in the absence of moisture or other chemically reactive
conditions, for at least a week.
The term "aliphatic" or "aliphatic group", as used herein, means a
straight-chain (i.e., unbranched) or branched, substituted or
unsubstituted hydrocarbon chain that is completely saturated or
that contains one or more units of unsaturation, or a monocyclic
hydrocarbon that is completely saturated or that contains one or
more units of unsaturation, but which is not aromatic (also
referred to herein as "carbocycle" "cycloaliphatic" or
"cycloalkyl"), that has a single point of attachment to the rest of
the molecule. In certain embodiments, aliphatic groups contain 1-6
aliphatic carbon atoms, and in yet other embodiments, aliphatic
groups contain 1-4 aliphatic carbon atoms. In some embodiments,
"cycloaliphatic" (or "carbocycle" or "cycloalkyl") refers to a
monocyclic C.sub.3-C.sub.6 hydrocarbon that is completely saturated
or that contains one or more units of unsaturation, but which is
not aromatic, that has a single point of attachment to the rest of
the molecule. Suitable aliphatic groups include, but are not
limited to, linear or branched, substituted or unsubstituted alkyl,
alkenyl, alkynyl groups and hybrids thereof such as
(cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.
The term "unsaturated", as used herein, means that a moiety has one
or more units of unsaturation.
The terms "haloalkyl", "haloalkenyl" and "haloalkoxy" means alkyl,
alkenyl or alkoxy, as the case may be, substituted with one or more
halogen atoms. The term "halogen" means F, Cl, Br, or I.
The term "aryl" used alone or as part of a larger moiety as in
"aralkyl", "aralkoxy", or "aryloxyalkyl", refers to monocyclic,
bicyclic and tricyclic ring systems having a total of five to
fourteen ring members, wherein at least one ring in the system is
aromatic and wherein each ring in the system contains 3 to 7 ring
members. The term "aryl" may be used interchangeably with the term
"aryl ring".
An aryl (including aralkyl, aralkoxy, aryloxyalkyl and the like) or
heteroaryl (including heteroaralkyl and heteroarylalkoxy and the
like) group may contain one or more substituents. Suitable
substituents on the unsaturated carbon atom of an aryl or
heteroaryl group are selected from halogen; R.sup.o; OR.sup.o;
SR.sup.o; 1,2-methylene-dioxy; 1,2-ethylene-dioxy; phenyl (Ph)
optionally substituted with R.sup.o; --O(Ph) optionally substituted
with R.sup.o; (CH.sub.2).sub.1-2(Ph), optionally substituted with
R.sup.o; CH.dbd.CH(Ph), optionally substituted with R.sup.o;
NO.sub.2; CN; N(R.sup.o).sub.2; NR.sup.oC(O)R.sup.o;
NR.sup.oC(O)N(R.sup.o).sub.2; NR.sup.oCO.sub.2R.sup.o;
--NR.sup.oNR.sup.oC(O)R.sup.o;
NR.sup.oNR.sup.oC(O)N(R.sup.o).sub.2;
NR.sup.oNR.sup.oCO.sub.2R.sup.o; C(O)C(O)R.sup.o;
C(O)CH.sub.2C(O)R.sup.o; CO.sub.2R.sup.o; C(O)R.sup.o;
C(O)N(R.sup.o).sub.2; OC(O)N(R.sup.o).sub.2; S(O).sub.2R.sup.o;
SO.sub.2N(R.sup.o).sub.2; S(O)R.sup.o;
NR.sup.oSO.sub.2N(R.sup.o).sub.2; NR.sup.oSO.sub.2R.sup.o;
C(.dbd.S)N(R.sup.o).sub.2; C(.dbd.NH)--N(R.sup.o).sub.2; or
(CH.sub.2).sub.0-2NHC(O)R.sup.o wherein each independent occurrence
of R.sup.o is selected from hydrogen, optionally substituted
C.sub.1-6 aliphatic, an unsubstituted 5-6 membered heteroaryl or
heterocyclic ring, phenyl, O(Ph), or CH.sub.2(Ph), or,
notwithstanding the definition above, two independent occurrences
of R.sup.o, on the same substituent or different substituents,
taken together with the atom(s) to which each R.sup.o group is
bound, form a 3-8 membered cycloalkyl, heterocyclyl, aryl, or
heteroaryl ring having 0-4 heteroatoms independently selected from
nitrogen, oxygen, or sulfur. Optional substituents on the aliphatic
group of R.sup.o are selected from NH.sub.2,
NH(C.sub.1-4aliphatic), N(C.sub.1-4aliphatic).sub.2, halogen,
C.sub.1-4aliphatic, OH, O(C.sub.1-4aliphatic), NO.sub.2, CN,
CO.sub.2H, CO.sub.2(C.sub.1-4aliphatic), O(haloC.sub.1-4
aliphatic), or haloC.sub.1-4 aliphatic, wherein each of the
foregoing C.sub.1-4aliphatic groups of R.sup.o is
unsubstituted.
An aliphatic or heteroaliphatic group or a non-aromatic
heterocyclic ring may contain one or more substituents. Suitable
substituents on the saturated carbon of an aliphatic or
heteroaliphatic group, or of a non-aromatic heterocyclic ring are
selected from those listed above for the unsaturated carbon of an
aryl or heteroaryl group and additionally include the following:
.dbd.O, .dbd.S, .dbd.NNHR*, .dbd.NN(R*).sub.2, .dbd.NNHC(O)R*,
.dbd.NNHCO.sub.2(alkyl), .dbd.NNHSO.sub.2(alkyl), or .dbd.NR, where
each R* is independently selected from hydrogen or an optionally
substituted C.sub.1-6 aliphatic. Optional substituents on the
aliphatic group of R* are selected from NH.sub.2, NH(C.sub.1-4
aliphatic), N(C.sub.1-4 aliphatic).sub.2, halogen,
C.sub.1-4aliphatic, OH, O(C.sub.1-4 aliphatic), NO.sub.2, CN,
CO.sub.2H, CO.sub.2(C.sub.1-4aliphatic), O(halo C.sub.1-4
aliphatic), or halo(C.sub.1-4 aliphatic), wherein each of the
foregoing C.sub.1-4aliphatic groups of R* is unsubstituted.
Optional substituents on the nitrogen of a non-aromatic
heterocyclic ring are selected from R.sup.+, N(R.sup.+).sub.2,
C(O)R.sup.+, CO.sub.2R.sup.+, C(O)C(O)R.sup.+,
C(O)CH.sub.2C(O)R.sup.+, SO.sub.2R.sup.+, SO.sub.2N
(R.sup.+).sub.2, C(.dbd.S)N(R.sup.+).sub.2,
C(.dbd.NH)--N(R.sup.+).sub.2, or NR.sup.+SO.sub.2R.sup.+; wherein
R.sup.+ is hydrogen, an optionally substituted C.sub.1-6 aliphatic,
optionally substituted phenyl, optionally substituted O(Ph),
optionally substituted CH.sub.2(Ph), optionally substituted
(CH.sub.2).sub.1-2(Ph); optionally substituted CH.dbd.CH(Ph); or an
unsubstituted 5-6 membered heteroaryl or heterocyclic ring having
one to four heteroatoms independently selected from oxygen,
nitrogen, or sulfur, or, notwithstanding the definition above, two
independent occurrences of R.sup.+, on the same substituent or
different substituents, taken together with the atom(s) to which
each R.sup.+ group is bound, form a 3-8-membered cycloalkyl,
heterocyclyl, aryl, or heteroaryl ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, or sulfur. Optional
substituents on the aliphatic group or the phenyl ring of R.sup.+
are selected from NH.sub.2, NH(C.sub.1-4 aliphatic), N(C.sub.1-4
aliphatic).sub.2, halogen, C.sub.1-4 aliphatic, OH, O(C.sub.1-4
aliphatic), NO.sub.2, CN, CO.sub.2H, CO.sub.2(C.sub.1-4 aliphatic),
O(halo C.sub.1-4 aliphatic), or halo(C.sub.1-4 aliphatic), wherein
each of the foregoing C.sub.1-4aliphatic groups of R.sup.+ is
unsubstituted.
Unless otherwise stated, structures depicted herein are also meant
to include all isomeric (e.g., enantiomeric, diastereomeric, and
geometric (or conformational)) forms of the structure; for example,
the R and S configurations for each asymmetric center, (Z) and (E)
double bond isomers, and (Z) and (E) conformational isomers.
Therefore, single stereochemical isomers as well as enantiomeric,
diastereomeric, and geometric (or conformational) mixtures of the
present compounds are within the scope of the invention. Unless
otherwise stated, all tautomeric forms of the compounds of the
invention are within the scope of the invention. Additionally,
unless otherwise stated, structures depicted herein are also meant
to include compounds that differ only in the presence of one or
more isotopically enriched atoms. For example, compounds having the
present structures except for the replacement of hydrogen by
deuterium or tritium, or the replacement of a carbon by a .sup.13C-
or .sup.14C-enriched carbon are within the scope of this invention.
Such compounds are useful, for example, as analytical tools or
probes in biological assays.
3. Description of Exemplary Compounds:
According to one embodiment, the present invention relates to a
compound of formula I wherein said compound is of formula Ia or
Ib:
##STR00003## or a pharmaceutically acceptable salt thereof, wherein
each m, R.sup.1, R.sup.2, and R.sup.3 group is as defined
above.
According to certain embodiments, the R.sup.1 moiety of any of
formulae I, Ia, and Ib, is C.sub.1-4 aliphatic optionally
substituted with --OR or --C.sub.1-3 haloalkyl. In certain
embodiments, the R.sup.1 moiety of any of formulae I, Ia, and Ib is
C.sub.1-4 aliphatic optionally substituted with --OH, --CH.sub.2F,
--CHF.sub.2, or --CF.sub.3. In other embodiments, the R.sup.1
moiety of any of formulae I, la, and lb is C.sub.1-4 aliphatic
optionally substituted with --OH. In yet other embodiments, R.sup.1
is unsubstituted.
According to another embodiment, the R.sup.1 moiety of any of
formulae I, Ia, and lb is isopropyl, 2-butyl, cyclopropyl, or
ethyl, wherein each moiety is optionally substituted with --OH,
--CHF.sub.2, --CH.sub.2F, or --CF.sub.3. In certain embodiments,
the R.sup.1 moiety of any of formulae I, Ia, and Ib is optionally
substituted with --OH or --CF.sub.3.
Another aspect of the present invention relates to a compound of
any of formulae I, Ia, and Ib wherein R.sup.2 is hydrogen,
C.sub.1-3 aliphatic, or chloro. According to yet another aspect,
the present invention relates to a compound of any of formulae I,
Ia, and Ib wherein R.sup.2 is chloro.
In certain embodiments, m is 1.
In other embodiments, the R.sup.3 moiety of any of formulae I, Ia,
and Ib is hydrogen, methyl, or chloro.
Representative compounds of formula I are set forth in Table 1
below.
TABLE-US-00001 TABLE 1 Examples of Compounds of Formula I:
##STR00004## I-1 ##STR00005## I-2 ##STR00006## I-3 ##STR00007## I-4
##STR00008## I-5 ##STR00009## I-6 ##STR00010## I-7 ##STR00011## I-8
##STR00012## I-9 ##STR00013## I-10 ##STR00014## I-11 ##STR00015##
I-12 ##STR00016## I-13 ##STR00017## I-14 ##STR00018## I-15
##STR00019## I-16 ##STR00020## I-17 ##STR00021## I-18
4. General Methods of Providing the Present Compounds:
The compounds of this invention may be prepared or isolated in
general by synthetic and/or pseudo-synthetic methods known to those
skilled in the art for analogous compounds and as illustrated by
the general Schemes I, II and III below and the preparative
examples that follow.
##STR00022## ##STR00023##
General Scheme I above shows a general method for preparing the
compounds of the present invention. At step (a), the pyrrole
compound 1 is iodinated and esterified to form 2. At step (b), the
pyrrole moiety is optionally protected at the --NH-- with a
suitable amino protecting group to form 3. Amino protecting groups
are well known in the art and are described in detail in Protecting
Groups in Organic Synthesis. Theodora W. Greene and Peter G. M.
Wuts, 1991, published by John Wiley and Sons, the entirety of which
is hereby incorporated by reference. The iodo moiety of compound 3
is displaced by an appropriate boronic acid or ester. As depicted
above, bis(pinacolato)diborane is used to form compound 4 however
other boronic esters or acids are amenable to this reaction and
would be apparent to one of ordinary skill in the art.
Because the present compounds relate to a multi-substituted
pyridine moiety, the order of reaction is considered and methods of
activating positions on the pyridine are utilized to direct the
regiochemistry. In step (d) above, the first leaving group L.sup.1
may be displaced by an alcohol, amine or thiol as desired. One of
ordinary skill in the art would recognize that various L.sup.1
leaving groups are amenable to this reaction. Examples of such
groups include, but are not limited to, halogen and activated
ethers. This reaction may be followed, at step (e), by the
replacement of a second leaving group L.sup.2 through either a
metal catalyzed coupling reaction or a nucleophilic displacement to
form compound 7. One of ordinary skill in the art would recognize
that various L.sup.2 leaving groups are amenable to this reaction.
Examples of such groups include, but are not limited to, halogen,
activated ethers, boronic acid, or boronic ester.
At step (f), the protecting group on the pyrrole moiety is removed
by methods suitable for removing the amino protecting group used.
Depending on which amino protecting group is used, the conditions
suitable for removing it may simultaneously saponify or otherwise
provide the carboxylate moiety as depicted above for compound 8. If
the conditions suitable for removing the amino protecting group are
not suitable for providing the carboxylate compound 8, then another
step may be employed. Compounds of formula 1 are prepared from 8 by
coupling the resulting carboxylate with a desired amine as depicted
at step (g). One of ordinary skill in the art would recognize that
a variety of conditions are useful for said coupling reaction and
can include the step of activating the carboxylate moiety of
compound 8 prior to or simultaneously with treatment with the
desired amine. Such conditions include, but are not limited to,
those described in detail in the Examples section below.
##STR00024##
Scheme II above depicts an alternate route to prepare intermediate
compound 6 useful for preparing compounds of the present invention.
At step (a), the N-oxide of compound 9 is prepared by treatment
with peroxide. The N-oxide compound 10 is then treated with nitric
acid to form the nitro compound 11. The L.sup.1 group of 11 is
displaced with the desired amine R.sup.1--NH.sub.2 to form 12 and
then the L.sup.2 group is introduced at step (d) to afford
intermediate 6. Compound 6 may then be utilized to prepare
compounds of the present invention according to the general Scheme
I above and the Examples provided below.
##STR00025##
Scheme III above shows an alternate method for preparing compound 7
from 6. In this method, the L.sup.2 group of the pyridinyl compound
6 is displaced by an appropriate boronic acid or ester derivative
to form 13, wherein R.sup.x and R.sub.y have the meanings as
defined for compounds of formula A infra. This boronate moiety is
then displaced by the L.sup.3 leaving group of the pyrrole depicted
above, wherein L.sup.3 is a suitable leaving group, to form
compound 7. Compound 7 is then used to prepare compounds of the
present invention by methods set forth above in Schemes I and II,
by those described in the Examples section and by methods known to
one of ordinary skill in the art.
One of skill in the art would recognize that a variety of compounds
of the present invention may be prepared according to the general
method of Schemes I, II and III, and the synthetic Examples set
forth below.
According to another embodiment, the present invention relates to a
compound of formula A:
##STR00026## or a salt thereof, wherein: PG is a suitable amino
protecting group; R.sup.z is a suitable carboxylate protecting
group; and R.sup.x and R.sup.y are independently hydrogen or
optionally substituted C.sub.1-6 aliphatic, or: R.sup.x and R.sup.y
are taken together to form an optionally substituted 5-7 membered
ring.
Suitable amino protecting groups are well known in the art and
include those described in detail in Protecting Groups in Organic
Synthesis, Theodora W. Greene and Peter G. M. Wuts, 1991, published
by John Wiley and Sons. In certain embodiments, the PG group of A
is an alkyl or aryl sulfonyl moiety. Examples of such groups
include mesyl, tosyl, nosyl, brosyl, and
2,4,6-trimethylbenzenesulfonyl ("Mts"). Other such groups include
Bn, PMB, Ms, Ts, SiR.sub.3, MOM, BOM, Tr, Ac, CO.sub.2R,
CH.sub.2OCH.sub.2CH.sub.2Si(CH.sub.3).sub.3.
Suitable carboxylate protecting groups are well known in the art
and are described in detail in Protecting Groups in Organic
Synthesis, Theodora W. Greene and Peter G. M. Wuts, 3.sup.rd
Edition, 1999, published by John Wiley and Sons. In certain
embodiments, the R.sup.z group of A is an optionally substituted
C.sub.1-6 aliphatic group or an optionally substituted aryl group.
Examples of suitable R.sup.z groups include methyl, ethyl, propyl,
isopropyl, butyl, isobutyl, benzyl, and phenyl wherein each group
is optionally substituted.
In certain embodiments, one or both of R.sup.x and R.sup.y are
hydrogen.
In other embodiments, R.sup.x and R.sup.y are taken together to
form an optionally substituted 5-6 membered ring. In yet other
embodiments, R.sup.x and R.sup.y are taken together to form a
4,4,5,5-tetramethyldioxaborolane moiety. Other suitable boronate
derivatives contemplated by the present invention include boronic
acid, B(O--C.sub.1-10 aliphatic).sub.2, and B(O-Aryl).sub.2.
According to yet another embodiment, the present invention provides
a compound of formula B:
##STR00027## or a salt thereof, wherein: R.sup.1 is a C.sub.1-6
aliphatic group, wherein R.sup.1 is optionally substituted with up
to 2 groups independently selected from --OR or --C.sub.1-3
haloalkyl; each R is independently hydrogen or C.sub.1-4 aliphatic;
R.sup.3 is hydrogen, C.sub.1-3 aliphatic, fluoro, or chloro; and
L.sup.2 is a suitable leaving group.
In certain embodiments, the present invention provides a compound
of formula B, as defined generally and in classes and subclasses
described above and herein, wherein L.sup.2 is not iodo when
R.sup.3 is chloro and R.sup.1 is isopropyl.
A suitable leaving group is a chemical group that is readily
displaced by a desired incoming chemical moiety. Thus, the choice
of the specific suitable leaving group is predicated upon its
ability to be readily displaced by the incoming chemical moiety of
formula A. Suitable leaving groups are well known in the art, e.g.,
see, "Advanced Organic Chemistry," Jerry March, 5.sup.th Ed., pp.
351-357, John Wiley and Sons, N.Y. Such leaving groups include, but
are not limited to, halogen, alkoxy, sulphonyloxy, optionally
substituted alkylsulphonyl, optionally substituted alkenylsulfonyl,
optionally substituted arylsulfonyl, and diazonium moieties.
Examples of suitable leaving groups include chloro, iodo, bromo,
fluoro, methanesulfonyl (mesyl), tosyl, triflate,
nitro-phenylsulfonyl (nosyl), and bromo-phenylsulfonyl (brosyl). In
certain embodiments, the L.sup.2 moiety of B is iodo.
According to an alternate embodiment, the suitable leaving group
may be generated in situ within the reaction medium. For example,
L.sup.2 in a compound of formula B may be generated in situ from a
precursor of that compound of formula B wherein said precursor
contains a group readily replaced by L.sup.2 in situ. In a specific
illustration of such a replacement, said precursor of a compound of
formula B contains a group (for example, a chloro group or hydroxyl
group) which is replaced in situ by L.sup.2, such as an iodo group.
The source of the iodo group may be, e.g., sodium iodide. Such an
in situ generation of a suitable leaving group is well known in the
art, e.g., see, "Advanced Organic Chemistry," Jerry March, pp.
430-431, 5.sup.th Ed., John Wiley and Sons, N.Y.
According to certain embodiments, the R.sup.1 moiety of formula B,
is C.sub.1-4 aliphatic optionally substituted with --OR or
--C.sub.1-3 haloalkyl. In certain embodiments, the R.sup.1 moiety
of formula B is C.sup.1-4 aliphatic optionally substituted with
--OH, --CH.sub.2F, --CHF.sub.2, or --CF.sub.3. In other
embodiments, the R.sup.1 moiety of formula B is C.sub.1-4 aliphatic
optionally substituted with --OH. In yet other embodiments, R.sup.1
is unsubstituted.
According to another embodiment, the R.sup.1 moiety of formula B is
isopropyl, 2-butyl, cyclopropyl, or ethyl, wherein each moiety is
optionally substituted with --OH or --CF.sub.3.
In other embodiments, the R.sup.3 moiety of formula B is hydrogen,
methyl, or chloro.
A compound of formula B may be prepared from a compound of formula
B':
##STR00028## or a salt thereof, wherein: R.sup.3 is hydrogen,
C.sub.1-3 aliphatic, fluoro, or chloro; and L.sup.1 and L.sup.2 are
each independently a suitable leaving group.
In certain embodiments, the present invention provides a compound
of formula B', as defined generally and in classes and subclasses
described above and herein, wherein L.sup.2 is not a boronate
moiety when R.sup.3 is chloro and R.sup.1 is fluoro.
In certain embodiments, a compound of B' is provided wherein
L.sup.2 is --B(OR.sup.x)(OR.sup.y). In other embodiments, one or
both of R.sup.x and R.sup.y are hydrogen. In other embodiments,
R.sup.x and R.sup.y are taken together to form an optionally
substituted 5-6 membered ring. In yet other embodiments, R.sup.x
and R.sup.y are taken together to form a
4,4,5,5-tetramethyldioxaborolane moiety.
As described above, a suitable leaving group is a chemical group
that is readily displaced by a desired incoming chemical moiety.
Suitable leaving groups are well known in the art, e.g., see,
"Advanced Organic Chemistry," Jerry March, 5.sup.th Ed., pp.
351-357, John Wiley and Sons, N.Y. Such leaving groups include, but
are not limited to, halogen, alkoxy, sulphonyloxy, optionally
substituted alkylsulphonyl, optionally substituted alkenylsulfonyl,
optionally substituted arylsulfonyl, and diazonium moieties.
Examples of suitable leaving groups include chloro, iodo, bromo,
fluoro, methanesulfonyl (mesyl), tosyl, triflate,
nitro-phenylsulfonyl (nosyl), and bromo-phenylsulfonyl (brosyl). In
certain embodiments, the L.sup.1 moiety of B' is halogen. In other
embodiment, the L.sup.1 moiety of B' is an optionally substituted
alkylsulphonyl, optionally substituted alkenylsulfonyl, or
optionally substituted arylsulfonyl group. In other embodiments,
the L.sup.1 moiety of B' is fluoro.
According to an alternate embodiment, the suitable leaving group
may be generated in situ within the reaction medium. For example,
L.sup.1 or L.sup.2 moieties in a compound of formula B'may be
generated in situ from a precursor of that compound of formula B'
wherein said precursor contains a group readily replaced by L.sup.1
or L.sup.2 in situ. Such an in situ generation of a suitable
leaving group is well known in the art, e.g., see, "Advanced
Organic Chemistry," Jerry March, pp. 430-431, 5.sup.th Ed., John
Wiley and Sons, N.Y.
According to another embodiment, the present invention provides a
method for preparing a compound of formula B:
##STR00029## or a salt thereof, comprising the step of reacting a
compound of formula B':
##STR00030## or a salt thereof, with a compound of formula
R.sup.1--NH.sub.2 wherein said reaction is performed in a suitable
medium and wherein: R.sup.1 is a C.sub.1-6 aliphatic group, wherein
R.sup.1 is optionally substituted with up to 2 groups independently
selected from --OR or --C.sub.1-3 haloalkyl; R is hydrogen or
C.sub.1-4 aliphatic; R.sup.3 is hydrogen, C.sub.1-3 aliphatic,
fluoro, or chloro; and L.sup.1 and L.sup.2 are each independently a
suitable leaving group.
In certain embodiments, said reaction is optionally performed in
the presence of a suitable base. One of ordinary skill would
recognize that the displacement of a leaving group by an amino
moiety is achieved either with or without the presence of a
suitable base. Such suitable bases are well known in the art and
include organic and inorganic bases.
A suitable medium is a solvent or a solvent mixture that, in
combination with the combined compounds, may facilitate the
progress of the reaction therebetween. The suitable solvent may
solubilize one or more of the reaction components, or,
alternatively, the suitable solvent may facilitate the agitation of
a suspension of one or more of the reaction components. Examples of
suitable solvents useful in the present invention are a protic
solvent, a halogenated hydrocarbon, an ether, an aromatic
hydrocarbon, a polar or a non-polar aprotic solvent, or any
mixtures thereof. Such mixtures include, for example, mixtures of
protic and non-protic solvents such as benzene/methanol/water;
benzene/water; DME/water, and the like.
These and other such suitable solvents are well known in the art,
e.g., see, "Advanced Organic Chemistry", Jerry March, 5.sup.th
edition, John Wiley and Sons, N.Y.
According to yet another embodiment, one or more reagents may
perform as the suitable solvent. For example, an organic base such
as triethylamine or diisopropylethylamine, if utilized in said
reaction, may serve as the solvent in addition to its role as a
basifying reagent.
In certain embodiments, the present invention provides a compound
of formula B' wherein R.sup.1 and R.sup.3 are as defined generally
and in classes and subclasses described above and herein.
According to another aspect, the present invention provides a
compound of formula C:
##STR00031## or a salt thereof, wherein: PG is a suitable amino
protecting group; R.sup.z is a suitable carboxylate protecting
group; R.sup.1 is a C.sub.1-6 aliphatic group, wherein R.sup.1 is
optionally substituted with up to 2 groups independently selected
from --OR or --C.sub.1-3 haloalkyl; each R is independently
hydrogen or C.sub.1-4 aliphatic; and R.sup.3 is hydrogen, C.sub.1-3
aliphatic, fluoro, or chloro.
As noted above, suitable amino protecting groups are well known in
the art and include those described in detail in Protecting Groups
in Organic Synthesis, Theodora W. Greene and Peter G. M. Wuts,
1991, published by John Wiley and Sons. In certain embodiments, the
PG group of C is an alkyl or aryl sulfonyl moiety. Examples of such
groups include mesyl, tosyl, nosyl, brosyl, and
2,4,6-trimethylbenzenesulfonyl ("Mts").
Suitable carboxylate protecting groups are well known in the art
and are described in detail in Protecting Groups in Organic
Synthesis, Theodora W. Greene and Peter G. M. Wuts, 1991, published
by John Wiley and Sons. In certain embodiments, the R.sup.z group
of C is an optionally substituted C.sub.1-6 aliphatic group or an
optionally substituted aryl group. Examples of suitable R.sup.z
groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl,
benzyl, and phenyl wherein each group is optionally
substituted.
According to certain embodiments, the R.sup.1 moiety of formula C,
is C.sub.1-4 aliphatic optionally substituted with --OR or
--C.sub.1-3 haloalkyl. In certain embodiments, the R.sup.1 moiety
of formula C is C.sub.1-4 aliphatic optionally substituted with
--OH, --CH.sub.2F, --CHF.sub.2, or --CF.sub.3. In other
embodiments, the R.sup.1 moiety of formula C is C.sub.1-4 aliphatic
optionally substituted with --OH. In yet other embodiments, R.sup.1
is unsubstituted.
According to another embodiment, the R.sup.1 moiety of formula C is
isopropyl, 2-butyl, cyclopropyl, or ethyl, wherein each moiety is
optionally substituted with --OH or --CF.sub.3.
In other embodiments, the R.sup.3 moiety of formula C is hydrogen,
methyl, or chloro.
Yet another aspect of the present invention relates to a method for
preparing a compound of formula C:
##STR00032## or a salt thereof, comprising the step of reacting a
compound of formula A:
##STR00033## or a salt thereof, with a compound of formula B:
##STR00034## or a salt thereof, wherein said reaction is performed
in a suitable medium and wherein: PG is a suitable amino protecting
group; L.sup.2 is a suitable leaving group. R.sup.z is a suitable
carboxylate protecting group; R.sup.x and R.sup.y are independently
hydrogen or optionally substituted C.sub.1-6 aliphatic, or: R.sup.x
and R.sup.y are taken together to form an optionally substituted
5-7 membered ring; R.sup.1 is a C.sub.1-6 aliphatic group, wherein
R.sup.1 is optionally substituted with up to 2 groups independently
selected from --OR or --C.sub.1-3 haloalkyl; each R is
independently hydrogen or C.sub.1-4 aliphatic; and R.sup.3 is
hydrogen, C.sub.1-3 aliphatic, fluoro, or chloro.
In certain embodiments, said reaction is performed in the presence
of Ni (II), Pd (O), or Pd(II) where each catalyst may be associated
with a ligand such as ferrocene or phosphine based ligands. In
other embodiments, said reaction is performed in the presence of
Pd(PPh.sub.3).sub.4. A suitable medium is a solvent or a solvent
mixture that, in combination with the combined compounds, may
facilitate the progress of the reaction therebetween. The suitable
solvent may solubilize one or more of the reaction components, or,
alternatively, the suitable solvent may facilitate the agitation of
a suspension of one or more of the reaction components. Examples of
suitable solvents useful in the present invention are a protic
solvent, a halogenated hydrocarbon, an ether, an aromatic
hydrocarbon, a polar or a non-polar aprotic solvent, or any
mixtures thereof. Such mixtures include, for example, mixtures of
protic and non-protic solvents such as benzene/methanol/water;
benzene/water; DME/water, and the like.
These and other such suitable solvents are well known in the art,
e.g., see, "Advanced Organic Chemistry", Jerry March, 5.sup.th
edition, John Wiley and Sons, N.Y.
In certain embodiments, the reaction between compounds A and B to
form C is performed in a mixture of DME and water.
In certain embodiments, the reaction between compounds A and B to
form C is performed at a temperature ranging from about 20.degree.
C. to 150.degree. C. In other embodiments, the reaction between
compounds A and B to form C is performed with microwave irradiation
at a temperature ranging from about 100.degree. C. to 250.degree.
C.
In still other embodiments, the reaction between compounds A and B
to form C is performed at a somewhat basic pH.
According to another embodiment, the present invention provides a
prodrug of a compound of formula 1 wherein said prodrug is of
formula II:
##STR00035## or a pharmaceutically acceptable salt thereof,
wherein: R.sup.1 is a C.sub.1-6 aliphatic group, wherein R.sup.1 is
optionally substituted with up to 2 groups independently selected
from --OR, --OR.sup.4, or --C.sub.1-3 haloalkyl; each R is
independently hydrogen or a C.sub.1-6 aliphatic; each R.sup.2 is
independently R, fluoro, or chloro; m is 0, 1, or 2; R.sup.3 is
hydrogen, C.sub.1-3 aliphatic, fluoro, or chloro; each R.sup.4 is
independently hydrogen, --C(R).sub.2O--R.sup.5, or R.sup.5,
provided that at least one R.sup.4 or R.sup.8 group is other than
hydrogen; each R.sup.5 is independently --C(O)R.sup.6,
--C(O)OR.sup.6, --C(O)-Q-R.sup.6,
--C(O)--(CH.sub.2).sub.n--C(O)OR.sup.6,
--C(O)--(CH.sub.2).sub.n--C(O)N(R.sup.7).sub.2,
--C(O)--(CH.sub.2).sub.n--CH(R.sup.6)N(R.sup.7).sub.2, --P(O)
(OR.sup.6).sub.2; each R.sup.6 is independently hydrogen, an
optionally substituted C.sub.1-6 aliphatic group or an optionally
substituted 5-8 membered saturated, partially unsaturated, or fully
unsaturated ring having 0-4 heteroatoms independently selected from
nitrogen, oxygen, or sulfur; each R.sup.7 is independently
hydrogen, --C(O)R.sup.6, --C(O)OR.sup.6, --S(O).sub.2R.sup.6,
--OR.sup.6, an optionally substituted C.sub.1-6 aliphatic group or
an optionally substituted 5-8 membered saturated, partially
unsaturated, or fully unsaturated ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, or sulfur, or: two
R.sup.6 on the same nitrogen atom taken together with the nitrogen
atom bound thereto form a 4-7 membered saturated, partially
unsaturated, or fully unsaturated ring having 1-3 heteroatoms in
addition to the nitrogen atom, independently selected from
nitrogen, oxygen, or sulfur; each n is 0-6; Q is an optionally
substituted C.sub.1-10 alkylidene chain wherein zero to four
methylene units of Q are independently replaced by --O--, --N(R)--,
--S--, --S(O)--, --S(O).sub.2--, or --C(O)--; and R.sup.8 is
hydrogen or --C(R).sub.2O--R.sup.5.
According to certain embodiments, the R.sup.1 moiety of formula II
is C.sub.1-4 aliphatic optionally substituted with --OR or
--C.sub.1-3 haloalkyl. In certain embodiments, the R.sup.1 moiety
of formula II is C.sub.1-4 aliphatic optionally substituted with
--OH, --CH.sub.2F, --CHF.sub.2, or --CF.sub.3. In other
embodiments, the R.sup.1 moiety of formula II is C.sub.1-4
aliphatic optionally substituted with --OH. In yet other
embodiments, R.sup.1 is unsubstituted.
According to another embodiment, the R.sup.1 moiety of formula II
is isopropyl, 2-butyl, cyclopropyl, or ethyl, wherein each moiety
is optionally substituted with --OH, --CHF.sub.2, --CH.sub.2F, or
--CF.sub.3. According to yet another embodiment, the R.sup.1 moiety
of formula II is isopropyl, 2-butyl, cyclopropyl, or ethyl, wherein
each moiety is optionally substituted with --OH or --CF.sub.3.
Another aspect of the present invention relates to a compound of
formula II wherein each R.sup.2 is independently hydrogen,
C.sub.1-3 aliphatic, or chloro. According to yet another aspect,
the present invention relates to a compound of formula II wherein
R.sup.2 is chloro and m is 1.
In other embodiments, the R.sup.3 moiety of formula II is hydrogen,
methyl, or chloro.
In certain embodiments, R.sup.4 is C(O)-Q-R.sup.6. Still other
embodiments related to a compound of formula II wherein R.sup.4 is
C(O)-Q-R.sup.6 and Q is an optionally substituted C.sub.1-8
alkylidene chain wherein zero to four methylene units of Q are
independently replaced by --O--, --N(R)--, --S--, --S(O)--,
--S(O).sub.2--, or --C(O)-- and R.sup.6 is as defined in general
and in classes and subclasses described above and herein. According
to another embodiment, Q is an optionally substituted C.sub.1-8
alkylidene chain wherein two to four methylene units of Q are
independently replaced by --O--. Such Q groups include
--CH.sub.2OCH.sub.2CH.sub.2O--,
--CH.sub.2OCH.sub.2CH.sub.2OCH.sub.2CH.sub.2O--, and the like.
Yet another aspect of the present invention provides a compound of
formula II wherein R.sup.4 is C(O)--(CH.sub.2).sub.n--CH
(R.sup.6)N(R.sup.7).sub.2. In certain embodiments, n is 0-2. In
other embodiments, R.sup.6 is an optionally substituted C.sub.1-6
aliphatic group. Examples of such R.sup.6 groups include methyl,
benzyl, ethyl, isopropyl, t-butyl, and the like. The R.sup.7 groups
of the C(O)--(CH.sub.2).sub.n--CH(R.sup.6)N(R.sup.7).sub.2 of
formula II include hydrogen and an optionally substituted C.sub.1-6
aliphatic group. Examples of such groups include methyl, benzyl,
ethyl, isopropyl, t-butyl, and the like.
According to one aspect, the present invention provides a compound
of formula II wherein R.sup.8 is hydrogen.
According to one embodiment, R.sup.4 is an L-valine ester.
In certain embodiments of the present invention, the R.sup.4 group
of formula II is --P(O)(OR.sup.6).sub.2. In other embodiments, each
R.sup.6 is independently hydrogen or an optionally substituted
C.sub.1-6 aliphatic group. Examples of such R.sup.6 groups include
methyl, benzyl, ethyl, isopropyl, t-butyl, and the like. In still
other embodiments, the R.sup.4 group of formula II is
--P(O)(OH).sub.2.
In certain embodiments, a compound of formula II provides
improvement with regard to one or more physical or physiological
characteristics. In other embodiments, a compound of formula II
imparts improvement with regard to one or more physical and
physiological characteristics.
Representative compounds of formula II are set forth in Table 2
below.
TABLE-US-00002 TABLE 2 ##STR00036## II-1 ##STR00037## II-2
##STR00038## II-3 ##STR00039## II-4 ##STR00040## II-5 ##STR00041##
II-6 ##STR00042## II-7 ##STR00043## II-8
Methods of preparing such prodrugs include those set Forth in
detail in the Examples section infra and methods known to one or
ordinary skill in the art.
5. Uses, Formulation and Administration
Pharmaceutically Acceptable Compositions
As discussed above, the present invention provides compounds that
are inhibitors of protein kinases, and thus the present compounds
are useful for the treatment of diseases, disorders, and conditions
including, but not limited to cancer, autoimmune disorders,
neurodegenerative and neurological disorders, schizophrenia,
bone-related disorders, liver disease, and cardiac disorders.
Accordingly, in another aspect of the present invention,
pharmaceutically acceptable compositions are provided, wherein
these compositions comprise any of the compounds as described
herein, and optionally comprise a pharmaceutically acceptable
carrier, adjuvant or vehicle. In certain embodiments, these
compositions optionally further comprise one or more additional
therapeutic agents.
It will also be appreciated that certain of the compounds of
present invention can exist in free form for treatment, or where
appropriate, as a pharmaceutically acceptable derivative thereof.
According to the present invention, a pharmaceutically acceptable
derivative includes, but is not limited to, pharmaceutically
acceptable salts, esters, salts of such esters, or any other adduct
or derivative which upon administration to a patient in need is
capable of providing, directly or indirectly, a compound as
otherwise described herein, or a metabolite or residue thereof.
As used herein, the term "pharmaceutically acceptable salt" refers
to those salts which are, within the scope of sound medical
judgment, suitable for use in contact with the tissues of humans
and lower animals without undue toxicity, irritation, allergic
response and the like, and are commensurate with a reasonable
benefit/risk ratio. A "pharmaceutically acceptable salt" means any
non-toxic salt or salt of an ester of a compound of this invention
that, upon administration to a recipient, is capable of providing,
either directly or indirectly, a compound of this invention or an
inhibitorily active metabolite or residue thereof. As used herein,
the term "inhibitorily active metabolite or residue thereof" means
that a metabolite or residue thereof is also an inhibitor of ERK2
protein kinase.
Pharmaceutically acceptable salts are well known in the art. For
example, S. M. Berge et al., describe pharmaceutically acceptable
salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19,
incorporated herein by reference. Pharmaceutically acceptable salts
of the compounds of this invention include those derived from
suitable inorganic and organic acids and bases. Examples of
pharmaceutically acceptable, nontoxic acid addition salts are salts
of an amino group formed with inorganic acids such as hydrochloric
acid, hydrobromic acid, phosphoric acid, sulfuric acid and
perchloric acid or with organic acids such as acetic acid, oxalic
acid, maleic acid, tartaric acid, citric acid, succinic acid or
malonic acid or by using other methods used in the art such as ion
exchange. Other pharmaceutically acceptable salts include adipate,
alginate, ascorbate, aspartate, benzenesulfonate, benzoate,
bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,
cyclopentanepropionate, digluconate, dodecylsulfate,
ethanesulfonate, formate, fumarate, glucoheptonate,
glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate,
hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate,
laurate, lauryl sulfate, malate, maleate, malonate,
methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate,
oleate, oxalate, palmitate, pamoate, pectinate, persulfate,
3-phenylpropionate, phosphate, picrate, pivalate, propionate,
stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, valerate salts, and the like.
Salts derived from appropriate bases include alkali metal, alkaline
earth metal, ammonium and N.sup.+(C.sub.1-4 alkyl).sub.4 salts.
This invention also envisions the quaternization of any basic
nitrogen-containing groups of the compounds disclosed herein. Water
or oil-soluble or dispersable products may be obtained by such
quaternization. Representative alkali or alkaline earth metal salts
include sodium, lithium, potassium, calcium, magnesium, and the
like. Further pharmaceutically acceptable salts include, when
appropriate, nontoxic ammonium, quaternary ammonium, and amine
cations formed using counterions such as halide, hydroxide,
carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and
aryl sulfonate.
As described above, the pharmaceutically acceptable compositions of
the present invention additionally comprise a pharmaceutically
acceptable carrier, adjuvant, or vehicle, which, as used herein,
includes any and all solvents, diluents, or other liquid vehicle,
dispersion or suspension aids, surface active agents, isotonic
agents, thickening or emulsifying agents, preservatives, solid
binders, lubricants and the like, as suited to the particular
dosage form desired. Remington's Pharmaceutical Sciences, Sixteenth
Edition, E. W. Martin (Mack Publishing Co., Easton, Pa., 1980)
discloses various carriers used in formulating pharmaceutically
acceptable compositions and known techniques for the preparation
thereof. Except insofar as any conventional carrier medium is
incompatible with the compounds of the invention, such as by
producing any undesirable biological effect or otherwise
interacting in a deleterious manner with any other component(s) of
the pharmaceutically acceptable composition, its use is
contemplated to be within the scope of this invention. Some
examples of materials which can serve as pharmaceutically
acceptable carriers include, but are not limited to, ion
exchangers, alumina, aluminum stearate, lecithin, serum proteins,
such as human serum albumin, buffer substances such as phosphates,
glycine, sorbic acid, or potassium sorbate, partial glyceride
mixtures of saturated vegetable fatty acids, water, salts or
electrolytes, such as protamine sulfate, disodium hydrogen
phosphate, potassium hydrogen phosphate, sodium chloride, zinc
salts, colloidal silica, magnesium trisilicate, polyvinyl
pyrrolidone, polyacrylates, waxes,
polyethylene-polyoxypropylene-block polymers, wool fat, sugars such
as lactose, glucose and sucrose; starches such as corn starch and
potato starch; cellulose and its derivatives such as sodium
carboxymethyl cellulose, ethyl cellulose and cellulose acetate;
powdered tragacanth; malt; gelatin; talc; excipients such as cocoa
butter and suppository waxes; oils such as peanut oil, cottonseed
oil; safflower oil; sesame oil; olive oil; corn oil and soybean
oil; glycols; such a propylene glycol or polyethylene glycol;
esters such as ethyl oleate and ethyl laurate; agar; buffering
agents such as magnesium hydroxide and aluminum hydroxide; alginic
acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl
alcohol, and phosphate buffer solutions, as well as other non-toxic
compatible lubricants such as sodium lauryl sulfate and magnesium
stearate, as well as coloring agents, releasing agents, coating
agents, sweetening, flavoring and perfuming agents, preservatives
and antioxidants can also be present in the composition, according
to the judgment of the formulator.
Uses of Compounds and Pharmaceutically Acceptable Compositions
In yet another aspect, a method for the treatment or lessening the
severity of cancer, an autoimmune disorder, a neurodegenerative or
neurological disorder, liver disease, or a cardiac disorder is
provided comprising administering an effective amount of a compound
of the present invention, or a pharmaceutically acceptable
composition comprising a compound of the present invention to a
subject in need thereof. In certain embodiments of the present
invention an "effective amount" of the compound or pharmaceutically
acceptable composition is that amount effective for treating or
lessening the severity of a disease, condition, or disorder
selected from cancer, an autoimmune disorder, a neurodegenerative
or neurological disorder, schizophrenia, a bone-related disorder,
liver disease, or a cardiac disorder. The compounds and
compositions, according to the method of the present invention, may
be administered using any amount and any route of administration
effective for treating or lessening the severity of cancer, an
autoimmune disorder, a neurodegenerative or neurological disorder,
schizophrenia, a bone-related disorder, liver disease, or a cardiac
disorder. The exact amount required will vary from subject to
subject, depending on the species, age, and general condition of
the subject, the severity of the infection, the particular agent,
its mode of administration, and the like. The compounds of the
invention are preferably formulated in dosage unit form for ease of
administration and uniformity of dosage. The expression "dosage
unit form" as used herein refers to a physically discrete unit of
agent appropriate for the patient to be treated. It will be
understood, however, that the total daily usage of the compounds
and compositions of the present invention will be decided by the
attending physician within the scope of sound medical judgment. The
specific effective dose level for any particular patient or
organism will depend upon a variety of factors including the
disorder being treated and the severity of the disorder; the
activity of the specific compound employed; the specific
composition employed; the age, body weight, general health, sex and
diet of the patient; the time of administration, route of
administration, and rate of excretion of the specific compound
employed; the duration of the treatment; drugs used in combination
or coincidental with the specific compound employed, and like
factors well known in the medical arts. The term "patient", as used
herein, means an animal, preferably a mammal, and most preferably a
human.
The pharmaceutically acceptable compositions of this invention can
be administered to humans and other animals orally, rectally,
parenterally, intracisternally, intravaginally, intraperitoneally,
topically (as by powders, ointments, or drops), bucally, as an oral
or nasal spray, or the like, depending on the severity of the
infection being treated. In certain embodiments, the compounds of
the invention may be administered orally or parenterally at dosage
levels of about 0.01 mg/kg to about 50 mg/kg and preferably from
about 1 mg/kg to about 25 mg/kg, of subject body weight per day,
one or more times a day, to obtain the desired therapeutic
effect.
Liquid dosage forms for oral administration include, but are not
limited to, pharmaceutically acceptable emulsions, microemulsions,
solutions, suspensions, syrups and elixirs. In addition to the
active compounds, the liquid dosage forms may contain inert
diluents commonly used in the art such as, for example, water or
other solvents, solubilizing agents and emulsifiers such as ethyl
alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl
alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol,
dimethylformamide, oils (in particular, cottonseed, groundnut,
corn, germ, olive, castor, and sesame oils), glycerol,
tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid
esters of sorbitan, and mixtures thereof. Besides inert diluents,
the oral compositions can also include adjuvants such as wetting
agents, emulsifying and suspending agents, sweetening, flavoring,
and perfuming agents.
Injectable preparations, for example, sterile injectable aqueous or
oleaginous suspensions may be formulated according to the known art
using suitable dispersing or wetting agents and suspending agents.
The sterile injectable preparation may also be a sterile injectable
solution, suspension or emulsion in a nontoxic parenterally
acceptable diluent or solvent, for example, as a solution in
1,3-butanediol. Among the acceptable vehicles and solvents that may
be employed are water, Ringer's solution, U.S.P. and isotonic
sodium chloride solution. In addition, sterile, fixed oils are
conventionally employed as a solvent or suspending medium. For this
purpose any bland fixed oil can be employed including synthetic
mono- or diglycerides. In addition, fatty acids such as oleic acid
are used in the preparation of injectables.
The injectable formulations can be sterilized, for example, by
filtration through a bacterial-retaining filter, or by
incorporating sterilizing agents in the form of sterile solid
compositions which can be dissolved or dispersed in sterile water
or other sterile injectable medium prior to use.
In order to prolong the effect of a compound of the present
invention, it is often desirable to slow the absorption of the
compound from subcutaneous or intramuscular injection. This may be
accomplished by the use of a liquid suspension of crystalline or
amorphous material with poor water solubility. The rate of
absorption of the compound then depends upon its rate of
dissolution that, in turn, may depend upon crystal size and
crystalline form. Alternatively, delayed absorption of a
parenterally administered compound form is accomplished by
dissolving or suspending the compound in an oil vehicle. Injectable
depot forms are made by forming microencapsule matrices of the
compound in biodegradable polymers such as
polylactide-polyglycolide. Depending upon the ratio of compound to
polymer and the nature of the particular polymer employed, the rate
of compound release can be controlled. Examples of other
biodegradable polymers include poly(orthoesters) and
poly(anhydrides). Depot injectable formulations are also prepared
by entrapping the compound in liposomes or microemulsions that are
compatible with body tissues.
Compositions for rectal or vaginal administration are preferably
suppositories which can be prepared by mixing the compounds of this
invention with suitable non-irritating excipients or carriers such
as cocoa butter, polyethylene glycol or a suppository wax which are
solid at ambient temperature but liquid at body temperature and
therefore melt in the rectum or vaginal cavity and release the
active compound.
Solid dosage forms for oral administration include capsules,
tablets, pills, powders, and granules. In such solid dosage forms,
the active compound is mixed with at least one inert,
pharmaceutically acceptable excipient or carrier such as sodium
citrate or dicalcium phosphate and/or a) fillers or extenders such
as starches, lactose, sucrose, glucose, mannitol, and silicic acid,
b) binders such as, for example, carboxymethylcellulose, alginates,
gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants
such as glycerol, d) disintegrating agents such as agar-agar,
calcium carbonate, potato or tapioca starch, alginic acid, certain
silicates, and sodium carbonate, e) solution retarding agents such
as paraffin, f) absorption accelerators such as quaternary ammonium
compounds, g) wetting agents such as, for example, cetyl alcohol
and glycerol monostearate, h) absorbents such as kaolin and
bentonite clay, and i) lubricants such as talc, calcium stearate,
magnesium stearate, solid polyethylene glycols, sodium lauryl
sulfate, and mixtures thereof. In the case of capsules, tablets and
pills, the dosage form may also comprise buffering agents.
Solid compositions of a similar type may also be employed as
fillers in soft and hard-filled gelatin capsules using such
excipients as lactose or milk sugar as well as high molecular
weight polyethylene glycols and the like. The solid dosage forms of
tablets, dragees, capsules, pills, and granules can be prepared
with coatings and shells such as enteric coatings and other
coatings well known in the pharmaceutical formulating art. They may
optionally contain opacifying agents and can also be of a
composition that they release the active ingredient(s) only, or
preferentially, in a certain part of the intestinal tract,
optionally, in a delayed manner. Examples of embedding compositions
that can be used include polymeric substances and waxes. Solid
compositions of a similar type may also be employed as fillers in
soft and hard-filled gelatin capsules using such excipients as
lactose or milk sugar as well as high molecular weight polethylene
glycols and the like.
The active compounds can also be in micro-encapsulated form with
one or more excipients as noted above. The solid dosage forms of
tablets, dragees, capsules, pills, and granules can be prepared
with coatings and shells such as enteric coatings, release
controlling coatings and other coatings well known in the
pharmaceutical formulating art. In such solid dosage forms the
active compound may be admixed with at least one inert diluent such
as sucrose, lactose or starch. Such dosage forms may also comprise,
as is normal practice, additional substances other than inert
diluents, e.g., tableting lubricants and other tableting aids such
a magnesium stearate and microcrystalline cellulose. In the case of
capsules, tablets and pills, the dosage forms may also comprise
buffering agents, They may optionally contain opacifying agents and
can also be of a composition that they release the active
ingredient(s) only, or preferentially, in a certain part of the
intestinal tract, optionally, in a delayed manner. Examples of
embedding compositions that can be used include polymeric
substances and waxes.
Dosage forms for topical or transdermal administration of a
compound of this invention include ointments, pastes, creams,
lotions, gels, powders, solutions, sprays, inhalants or patches.
The active component is admixed under sterile conditions with a
pharmaceutically acceptable carrier and any needed preservatives or
buffers as may be required. Ophthalmic formulation, ear drops, and
eye drops are also contemplated as being within the scope of this
invention. Additionally, the present invention contemplates the use
of transdermal patches, which have the added advantage of providing
controlled delivery of a compound to the body. Such dosage forms
can be made by dissolving or dispensing the compound in the proper
medium. Absorption enhancers can also be used to increase the flux
of the compound across the skin. The rate can be controlled by
either providing a rate controlling membrane or by dispersing the
compound in a polymer matrix or gel.
As described generally above, the compounds of the invention are
useful as inhibitors of ERK protein kinases. In one embodiment, the
compounds and compositions of the invention are inhibitors of one
or both of ERK1 and ERK2 protein kinases and thus, without wishing
to be bound by any particular theory, the compounds and
compositions are particularly useful for treating or lessening the
severity of a disease, condition, or disorder where activation of
one or both of ERK1 and ERK2 protein kinases is implicated in the
disease, condition, or disorder. When activation of ERK1 and/or
ERK2 protein kinases is implicated in a particular disease,
condition, or disorder, the disease, condition, or disorder may
also be referred to as "ERK1- or ERK2-mediated disease", condition,
or disease symptom. Accordingly, in another aspect, the present
invention provides a method for treating or lessening the severity
of a disease, condition, or disorder where activation of one or
both of ERK1 and ERK2 protein kinases is implicated in said
disease, condition, or disorder.
The activity of a compound utilized in this invention as an
inhibitor of ERK1 and/or ERK2 protein kinases may be assayed in
vitro, in vivo or in a cell line. In vitro assays include assays
that determine inhibition of either the phosphorylation activity or
ATPase activity of activated ERK1 or ERK2 protein kinases.
Alternate in vitro assays quantitate the ability of the inhibitor
to bind to ERK1 or ERK2 protein kinases. Inhibitor binding may be
measured by radiolabelling the inhibitor prior to binding,
isolating the inhibitor/ERK1 or inhibitor/ERK2 complex and
determining the amount of radiolabel bound. Alternatively,
inhibitor binding may be determined by running a competition
experiment where new inhibitors are incubated with ERK1 or ERK2
protein kinases bound to known radioligands.
The term "measurably inhibit", as used herein means a measurable
change in ERK1 or ERK2 protein kinase activity between a sample
comprising said composition and a ERK1 or ERK2 protein kinase and
an equivalent sample comprising ERK1 or ERK2 protein kinase in the
absence of said composition. Such measurements of protein kinase
activity are known to one of ordinary skill in the art and include
those methods set forth herein below.
According to another embodiment, the invention relates to a method
of inhibiting ERK1 or ERK2 protein kinase activity in a patient
comprising the step of administering to said patient a compound of
the present invention, or a composition comprising said
compound.
The term "ERK-mediated condition" or "disease", as used herein,
means any disease or other deleterious condition in which ERK is
known to play a role. The term "ERK-mediated condition" or
"disease" also means those diseases or conditions that are
alleviated by treatment with an ERK inhibitor. Such conditions
include, without limitation, cancer, stroke, diabetes,
hepatomegaly, cardiovascular disease including cardiomegaly,
Alzheimer's disease, cystic fibrosis, viral disease, autoimmune
diseases, atherosclerosis, restenosis, psoriasis, allergic
disorders including asthma, inflammation, neurological disorders
and hormone-related diseases. The term "cancer" includes, but is
not limited to the following cancers: breast, ovary, cervix,
prostate, testis, genitourinary tract, esophagus, larynx,
glioblastoma, neuroblastoma, stomach, skin, keratoacanthoma, lung,
epidermoid carcinoma, large cell carcinoma, small cell carcinoma,
lung adenocarcinoma, bone, colon, adenoma, pancreas,
adenocarcinoma, thyroid, follicular carcinoma, undifferentiated
carcinoma, papillary carcinoma, seminoma, melanoma, sarcoma,
bladder carcinoma, liver carcinoma and biliary passages, kidney
carcinoma, myeloid disorders, lymphoid disorders, Hodgkin's, hairy
cells, buccal cavity and pharynx (oral), lip, tongue, mouth,
pharynx, small intestine, colon-rectum, large intestine, rectum,
brain and central nervous system, and leukemia.
Accordingly, another embodiment of the present invention relates to
treating or lessening the severity of one or more diseases in which
ERK is known to play a role. Specifically, the present invention
relates to a method of treating or lessening the severity of a
disease or condition selected from cancer, stroke, diabetes,
hepatomegaly, cardiovascular disease including cardiomegaly,
Alzheimer's disease, cystic fibrosis, viral disease, autoimmune
diseases, atherosclerosis, restenosis, psoriasis, allergic
disorders including asthma, inflammation, neurological disorders
and hormone-related diseases, wherein said method comprises
administering to a patient in need thereof a composition according
to the present invention.
According to another embodiment, the present invention relates to a
method of treating a cancer selected from breast, ovary, cervix,
prostate, testis, genitourinary tract, esophagus, larynx,
glioblastoma, neuroblastoma, stomach, skin, keratoacanthoma, lung,
epidermoid carcinoma, large cell carcinoma, small cell carcinoma,
lung adenocarcinoma, bone, colon, adenoma, pancreas,
adenocarcinoma, thyroid, follicular carcinoma, undifferentiated
carcinoma, papillary carcinoma, seminoma, melanoma, sarcoma,
bladder carcinoma, liver carcinoma and biliary passages, kidney
carcinoma, myeloid disorders, lymphoid disorders, Hodgkin's, hairy
cells, buccal cavity and pharynx (oral), lip, tongue, mouth,
pharynx, small intestine, colon-rectum, large intestine, rectum,
brain and central nervous system, and leukemia.
Another embodiment relates to a method of treating melanoma, breast
cancer, colon cancer, or pancreatic cancer in a patient in need
thereof.
It will also be appreciated that the compounds and pharmaceutically
acceptable compositions of the present invention can be employed in
combination therapies, that is, the compounds and pharmaceutically
acceptable compositions can be administered concurrently with,
prior to, or subsequent to, one or more other desired therapeutics
or medical procedures. The particular combination of therapies
(therapeutics or procedures) to employ in a combination regimen
will take into account compatibility of the desired therapeutics
and/or procedures and the desired therapeutic effect to be
achieved. It will also be appreciated that the therapies employed
may achieve a desired effect for the same disorder (for example, an
inventive compound may be administered concurrently with another
agent used to treat the same disorder), or they may achieve
different effects (e.g., control of any adverse effects). As used
herein, additional therapeutic agents that are normally
administered to treat or prevent a particular disease, or
condition, are known as "appropriate for the disease, or condition,
being treated".
For example, chemotherapeutic agents or other anti-proliferative
agents may be combined with the compounds of this invention to
treat proliferative diseases and cancer. Examples of known
chemotherapeutic agents include, but are not limited to, For
example, other therapies or anticancer agents that may be used in
combination with the inventive anticancer agents of the present
invention include surgery, radiotherapy (in but a few examples,
gamma.-radiation, neutron beam radiotherapy, electron beam
radiotherapy, proton therapy, brachytherapy, and systemic
radioactive isotopes, to name a few), endocrine therapy, biologic
response modifiers (interferons, interleukins, and tumor necrosis
factor (TNF) to name a few), hyperthermia and cryotherapy, agents
to attenuate any adverse effects (e.g., antiemetics), and other
approved chemotherapeutic drugs, including, but not limited to,
alkylating drugs (mechlorethamine, chlorambucil, Cyclophosphamide,
Melphalan, Ifosfamide), antimetabolites (Methotrexate), purine
antagonists and pyrimidine antagonists (6-Mercaptopurine,
5-Fluorouracil, Cytarabile, Gemcitabine), spindle poisons
(Vinblastine, Vincristine, Vinorelbine, Paclitaxel),
podophyllotoxins (Etoposide, Irinotecan, Topotecan), antibiotics
(Doxorubicin, Bleomycin, Mitomycin), nitrosoureas (Carmustine,
Lomustine), inorganic ions (Cisplatin, Carboplatin), enzymes
(Asparaginase), and hormones (Tamoxifen, Leuprolide, Flutamide, and
Megestrol), Gleevec.TM., adriamycin, dexamethasone, and
cyclophosphamide. For a more comprehensive discussion of updated
cancer therapies see, http://www.nci.nih.gov/, a list of the FDA
approved oncology drugs at
http://www.fda.gov/cder/cancer/druglistframe.htm, and The Merck
Manual, Seventeenth Ed. 1999, the entire contents of which are
hereby incorporated by reference.
Other examples of agents the inhibitors of this invention may also
be combined with include, without limitation: treatments for
Alzheimer's Disease such as Aricept.RTM. and Excelon.RTM.;
treatments for Parkinson's Disease such as L-DOPA/carbidopa,
entacapone, ropinrole, pramipexole, bromocriptine, pergolide,
trihexephendyl, and amantadine; agents for treating Multiple
Sclerosis (MS) such as beta interferon (e.g., Avonex.RTM. and
Rebif.RTM.), Copaxone.RTM., and mitoxantrone; treatments for asthma
such as albuterol and Singulair.RTM.; agents for treating
schizophrenia such as zyprexa, risperdal, seroquel, and
haloperidol; anti-inflammatory agents such as corticosteroids, TNF
blockers, IL-1 RA, azathioprine, cyclophosphamide, and
sulfasalazine; immunomodulatory and immunosuppressive agents such
as cyclosporin, tacrolimus, rapamycin, mycophenolate mofetil,
interferons, corticosteroids, cyclophosphamide, azathioprine, and
sulfasalazine; neurotrophic factors such as acetylcholinesterase
inhibitors, MAO inhibitors, interferons, anti-convulsants, ion
channel blockers, riluzole, and anti-Parkinsonian agents; agents
for treating cardiovascular disease such as beta-blockers, ACE
inhibitors, diuretics, nitrates, calcium channel blockers, and
statins; agents for treating liver disease such as corticosteroids,
cholestyramine, interferons, and anti-viral agents; agents for
treating blood disorders such as corticosteroids, anti-leukemic
agents, and growth factors; and agents for treating
immunodeficiency disorders such as gamma globulin.
The amount of additional therapeutic agent present in the
compositions of this invention will be no more than the amount that
would normally be administered in a composition comprising that
therapeutic agent as the only active agent. Preferably the amount
of additional therapeutic agent in the presently disclosed
compositions will range from about 50% to 100% of the amount
normally present in a composition comprising that agent as the only
therapeutically active agent.
In an alternate embodiment, the methods of this invention that
utilize compositions that do not contain an additional therapeutic
agent, comprise the additional step of separately administering to
said patient an additional therapeutic agent. When these additional
therapeutic agents are administered separately they may be
administered to the patient prior to, sequentially with or
following administration of the compositions of this invention.
The compounds of this invention or pharmaceutically acceptable
compositions thereof may also be incorporated into compositions for
coating implantable medical devices, such as prostheses, artificial
valves, vascular grafts, stents and catheters. Accordingly, the
present invention, in another aspect, includes a composition for
coating an implantable device comprising a compound of the present
invention as described generally above, and in classes and
subclasses herein, and a carrier suitable for coating said
implantable device. In still another aspect, the present invention
includes an implantable device coated with a composition comprising
a compound of the present invention as described generally above,
and in classes and subclasses herein, and a carrier suitable for
coating said implantable device.
Vascular stents, for example, have been used to overcome restenosis
(re-narrowing of the vessel wall after injury). However, patients
using stents or other implantable devices risk clot formation or
platelet activation. These unwanted effects may be prevented or
mitigated by pre-coating the device with a pharmaceutically
acceptable composition comprising a kinase inhibitor. Suitable
coatings and the general preparation of coated implantable devices
are described in U.S. Pat. Nos. 6,099,562; 5,886,026; and
5,304,121. The coatings are typically biocompatible polymeric
materials such as a hydrogel polymer, polymethyldisiloxane,
polycaprolactone, polyethylene glycol, polylactic acid, ethylene
vinyl acetate, and mixtures thereof. The coatings may optionally be
further covered by a suitable topcoat of fluorosilicone,
polysaccarides, polyethylene glycol, phospholipids or combinations
thereof to impart controlled release characteristics in the
composition.
Another aspect of the invention relates to inhibiting ERK1 or ERK2
protein kinase activity in a biological sample or a patient, which
method comprises administering to the patient, or contacting said
biological sample with a compound of the present invention or a
composition comprising said compound. The term "biological sample",
as used herein, includes, without limitation, cell cultures or
extracts thereof; biopsied material obtained from a mammal or
extracts thereof; and blood, saliva, urine, feces, semen, tears, or
other body fluids or extracts thereof.
Inhibition of ERK1 or ERK2 protein kinase activity in a biological
sample is useful for a variety of purposes that are known to one of
skill in the art. Examples of such purposes include, but are not
limited to, blood transfusion, organ-transplantation, biological
specimen storage, and biological assays.
SYNTHETIC EXAMPLES
As used herein, the term "R.sub.t" refers to the HPLC retention
time, in minutes, associated with the compound. Unless otherwise
indicated, the HPLC method utilized to obtain the reported
retention time is as follows: Column: Agilent/ZORBAX SB--C18/5
.mu.m/3.0.times.150 mm/PN 883975-302/SN USBM001410 Gradient: 10-90%
MeCN over 8 minutes Flow: 1.0 mL/minute Detection: 214 nm and 254
nm
Unless otherwise indicated, each .sup.1H NMR was obtained at 500
MHz in CDCl.sub.3 and compound numbers correspond to those compound
numbers recited in Table 1.
Example 1
Compound 1-9 was prepared as follows:
##STR00044##
2,2,2-Trichloro-1-(4-iodo-1H-pyrrol-2-yl)ethanone: To a stirred
solution of 50 g (235 mmol, 1.0 equiv.) of
2,2,2-trichloro-1-(1H-pyrrol-2-yl)-ethanone in dry dichloromethane
(400 mL) under nitrogen, a solution of iodine monochloride (39 g,
240 mmol, 1.02 equivalents) in of dichloromethane (200 mL) was
added dropwise. The resulting mixture was stirred at room
temperature for 2 hours. The solution was washed with 10% potassium
carbonate, water, 1.0 M sodium thiosulfate, saturated sodium
chloride, dried over sodium sulfate, filtered, and concentrated
under reduced pressure. The solid was recrystallized from
hexanes/methyl acetate to afford the title compound (68.5 g, 86%)
as a colorless solid (86%). MS FIA: 335.8, 337.8 ES-.
4-Iodo-1H-pyrrole-2-carboxylic acid methyl ester: To a stirred
solution of 2,2,2-trichloro-1-(4-iodo-1H-pyrrol-2-yl) ethanone (68
g, 201 mmol, 1.0 equivalent) in dry methanol (400 mL) under
nitrogen, was added a solution of sodium methoxide in methanol
(4.37 M, 54 mL, 235 mmol, 1.2 equivalents) over 10 minutes. The
resulting mixture was stirred at room temperature for 1 hour. The
volatiles were removed under reduced pressure and the crude was
then partitioned between water and tert-butylmethyl ether. The
organic phase was separated, washed two times with water, saturated
sodium chloride, dried over sodium sulfate, filtered and
concentrated under vacuum to afford the title compound (48 g, 96%)
as a colorless solid, that was used directly without further
purification.
4-Iodo-1-(toluene-4-sulfonyl)-1H-pyrrole-2-carboxylic acid methyl
ester: 4-Iodo-1H-pyrrole-2-carboxylic acid methyl ester (24.6 g, 98
mmol, 1.0 equivalent) was dissolved in dichloromethane (150 mL) and
triethylamine (30 mL, 215.6 mmol, 2.2 equivalents).
4-(Dimethylamino)pyridine (1.2 g, 9.8 mmol, 0.1 equivalent) and
p-toluenesulfonylchloride (20.6 g, 107.8 mmol, 1.1 equivalents)
were added and the reaction mixture was stirred for 16 hours at
room temperature. The reaction was quenched with 1 M HCl and the
organic layer was washed with aqueous sodium bicarbonate and brine.
After drying over magnesium sulfate, the solvent was removed under
reduced pressure and the residue was crystallized from
tert-butylmethyl ether, yielding the title compound as a pale
yellow solid (30 g, 75%). R.sub.t(min) 8.259 minutes.
4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-1-(toluene-4-sulfonyl)-1-
H-pyrrole-2-carboxylic acid methyl ester: To a degassed solution of
4-iodo-1-(toluene-4-sulfonyl)-1H-pyrrole-2-carboxylic acid methyl
ester (20 g, 49.4 mmol, 1.0 equivalent) and bis(pinacolato)diborane
(15 g, 65 mmol, 1.3 equivalents) in DMF (200 mL) under nitrogen,
was added dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium
(II) dichloromethane adduct (3.6 g, 4.9 mmol, 0.1 equivalent). The
reaction mixture was then stirred at 80.degree. C. for 18 hours.
After removing the DMF under reduced pressure, the resulting thick
oil residue was suspended in diethyl ether (500 mL) and a solid
precipitated immediately. This solid was removed by filtration and
the filtrate was washed with 1M HCl, water, brine and dried over
MgSO.sub.4. Concentration afforded the title compound as a white
solid and used without further purification (10 g, 50%). LC/MS:
R.sub.t(min) 4.6; 406.4 ES+. MS FIA: 406.2 ES+. .sup.1HNMR .delta.
1.2 (s, 12H), 2.35 (s, 3H), 3.8 (s, 3H), 7.2 (m, 3H), 7.8 (d, 2H),
8.0 (s, 1H).
N,N'-2-(5-Chloro-4-iodo-pyridyl)-isopropylamine:
Method A. (Microwave)
In a 10 mL microwave tube, 5-chloro-2-fluoro-4-iodopyridine (1.0 g,
3.9 mmol, 1.0 equivalent) was dissolved in DMSO (4.0 mL) and then
ispropylamine (0.99 mL, 11.7 mmol, 3.0 equivalents) was added. The
tube was sealed and placed under microwave irradiation for 600 sec
at 150.degree. C.
This reaction was repeated six times. The reaction mixtures were
combined, then diluted in ethyl acetate and washed with water.
After drying over sodium sulfate, the solvent was evaporated to
afford the title compound as a thick brown oil (5.6 g, 80%) which
was used directly without further purification. R.sub.t(min) 4.614;
MS FIA: 296.9 ES+. .sup.1HNMRsssssss .delta. 1.25 (d, 6H), 3.65 (m,
1H), 7.15 (s, 1H), 7.75 (s, 1H).
Method B: (Thermal)
5-Chloro-2-fluoro-4-iodopyridine (400 mg, 1.55 mmol, 1.0
equivalent) was dissolved in ethanol (5.0 mL) and then
isopropylamine (0.66 mL, 7.8 mmol, 5.0 equivalents) was added. The
resulting solution was stirred at 80.degree. C. for 48 hours. The
reaction mixture was then diluted in ethyl acetate and washed with
water. After drying over sodium sulfate, the solvent was evaporated
and a thick brown oil was obtained, which was then purified by
flash chromatography on silica gel eluting with mixtures of
hexanes/ethyl acetate (from 99:1 to 80:20) to afford the title
compound as a pale yellow solid (96 mg, 21%).
4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1-(toluene-4-sulfonyl)-1H-pyrro-
le-2-carboxylic acid methyl ester: To a solution of
N,N'-2-(5-chloro-4-iodo-pyridyl)-isopropylamine (0.53 g, 1.8 mmol,
1.0 equivalent) and
4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1-(toluene-4-sulfonyl)-1-
H-pyrrole-2-carboxylic acid methyl ester (0.78 g, 1.8 mmol, 1.0
equivalent) in DME (4.0 mL) was added a solution of aqueous 2 M
sodium carbonate (1.0 mL) followed by Pd(PPh.sub.3).sub.4 (0.21 mg,
0.18 mmol, 0.1 equivalent). The microwave tube was sealed and the
reaction mixture was irradiated by microwave for 1800 sec. at
170.degree. C. The crude of six reactions were combined and diluted
in ethyl acetate and washed with water. After drying the organic
layer with sodium sulfate, the solvent was removed and the
resulting thick oil was adsorbed on silica gel. The crude was then
purified by flash chromatography on silica, eluting with
hexanes/ethyl acetate mixtures (from 99:1 to 70:30) to afford the
title compound as a yellow solid (3.1 g, 61% over two steps).
R.sub.t(min) 6.556. MS FIA: 448.1 ES+. .sup.1HNMR .delta. 1.45 (d,
6H), 2.5 (s, 3H), 3.81 (s, 3H), 6.8 (s, 1H), 7.35 (s, 1H), 7.4 (d,
2H), 8.0 (m, 3H), 8.3 (s, 1H).
4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1-(2,4,6-trimethylbenzenesulfon-
yl)-1H-pyrrole-2-carboxylic acid methyl ester: To a solution of
N,N'-2-(5-chloro-4-iodopyridyl)-isopropylamine (96 mg, 0.32 mmol,
1.0 equivalent) and
4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1-(2,4,6-trimethylbenzen-
esulfonyl)-1H-pyrrole-2-carboxylic acid methyl ester (152 mg, 0.35
mmol, 1.1 equivalents) in DME (2 mL), was added a solution of
aqueous 2 M sodium carbonate (0.2 mL) followed by
Pd(PPh.sub.3).sub.4 (37 mg, 0.032 mmol, 0.1 equivalent). The
reaction mixture was stirred at 80.degree. C. for 16 hours. The
crude was diluted in ethyl acetate and washed with water. After
drying the organic layer with sodium sulfate, the solvent was
removed and the resulting thick oil was adsorbed on silica gel. The
crude was then purified by flash chromatography on silica, eluting
with hexanes/ethyl acetate mixtures (from 99:1 to 80:20) to afford
the title compound as a yellow solid (65 mg, 43%). R.sub.t(min)
7.290. MS FIA:476.1 ES+.
4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic
acid:
Method A. (Microwave)
A solution of
4-(5-chloro-2-isopropylaminopyridin-4-yl)-1-(toluene-4-sulfonyl)-1H-pyrro-
le-2-carboxylic acid methyl ester (3.1 g, 6.9 mmol, 1.0 equivalent)
in THF (2.0 mL) was added to a solution of lithium hydroxide
monohydrated (710 mg, 17.3 mmol, 2.5 equivalents) in water (3.0
mL). The microwave tube was sealed and the reaction mixture was
irradiated by microwave for 1200 sec. at 150.degree. C. The crude
solution was acidified with aqueous 6N HCl. The solvent was
evaporated off to afford the title compound which was used directly
without further purification. R.sub.t(min): 3.574. FIA MS: 279.9
ES+; 278.2 ES-.
Method B: (Thermal)
A solution of
4-(5-chloro-2-isopropylaminopyridin-4-yl)-1-(2,4,6-trimethylbenzenesulfon-
yl)-1H-pyrrole-2-carboxylic acid methyl ester (0.69 g, 1.4 mmol,
1.0 equivalent) in THF (3.0 mL) was added to a solution of lithium
hydroxide monohydrated (1.19 g, 29 mmol, 20.0 equivalents) in water
(3.0 mL). The mixture was then refluxed for 8 hours. The crude
solution was acidified with aqueous 6N HCl until cloudy, the
organic solvent was partially removed and the product precipitated.
The title compound was isolated by filtration and washed with water
and diethyl ether, yielding a white solid (0.38 g, 96%).
4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic
acid [1-(3-chlorophenyl)-2-hydroxyethyl]amide: To a suspension of
4-(5-chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic
acid (1.93 g, 6.9 mmol, 1.0 equivalent) in DMF (5.0 mL) was added
EDCI (1.45 g, 7.6 mmol, 1.1 equivalents), HOBt (0.94 g, 6.9 mmol,
1.0 equivalent) and (S)-3-chlorophenylglycynol (1.58 g, 7.6 mmol,
1.1 equivalents). Diisopropylethylamine (2.7 mL) was then added and
the resulting mixture was stirred a room temperature overnight. The
mixture was then poured into water and extracted with ethyl
acetate. After drying over sodium sulfate, the solvent was removed
and the crude was adsorbed on silica gel. Purification was effected
by flash chromatography on silica, eluting with mixtures of
hexanes/acetone (from 80:20 to 60:40) to afford the title compound
as white solid (1.9 g, 64%). R.sub.t(min) 4.981 s. FIA MS: 433.1
ES+; 431.2 ES-. .sup.1HNMR (CD.sub.3OD) .delta. 1.31 (d, 6H), 3.85
(m, 3H), 5.15 (t, 1H), 7.01 (s, 1H), 7.25 (m, 3H), 7.4 (s, 1H),
7.45 (s, 1H), 7.7 (s, 1H), 7.95 (s, 1H).
Example 2
Compound 1-9 was also prepared according to following alternate
method:
##STR00045##
2,5-Dichloro-4-nitropyridine N-oxide: To a suspension of
2-chloro-5-chloropyridine (10 g, 0.067 mol) in acetic anhydride (25
mL) was added hydrogen peroxide 30% (25 mL) in small portions. This
mixture was stirred at room temperature for 24 hours and then
heated at 60.degree. C. for 30 hours. After removing the excess of
acetic acid under reduced pressure, the residue was added in small
portions to concentrated sulfuric acid (15 mL). The resulting
solution was added to a mixture of concentrated sulfuric acid (15
mL) and fuming nitric acid (25 mL) and then heated at 100.degree.
C. for 90 minutes. The reaction mixture was poured on ice,
neutralized with solid ammonium carbonate and finally with aqueous
ammonia until a basic pH was obtained and A precipitate formed. The
precipitate was collected by filtration to afford the title
compound as a pale yellow solid (3.1 g), R.sub.t(min) 3.75. MS FIA
shows no peak. .sup.1HNMR (DMSO-d.sub.6) .delta. 8.78 (s, 1H), 9.15
(s, 1H).
4-Bromo-2-chloro-5-N-isopropylpyridin-2-amine N-oxide: To
2,5-dichloro-4-nitropyridine N-oxide (400 mg, 1.9 mmol) was added
acetyl bromide (2 mL) very slowly. The reaction mixture was then
heated at 80.degree. C. for 10 minutes. The solvent was removed
under a stream of nitrogen and the crude product was dried under
high vacuum. The crude material (165 mg, 0.62 mmol) was dissolved
in ethanol (2 mL), iso-propylamine (0.53 mL) added and the
resulting mixture was heated at 80.degree. C. for 2 hours. The
crude solution was then purified by reversed phase HPLC
(acetonitrile/water/TFA 1%) to afford the title compound as a pale
yellow solid (60 mg, 36.6%). R.sub.t(min) 5.275. MS FIA264.8, 266.9
ES+.
4-(5-chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic
acid [1-(3-chlorophenyl)-2-hydroxyethyl]amide (1-9):
4-Bromo-2-chloro-5-N-isopropylpyridin-2-amine N-oxide (25 mg, 0.094
mmol, 1.0 equivalent) and
4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1-(2,4,6-trimethylbenzen-
sulfonyl)-1H-pyrrole-2-carboxylic acid methyl ester (39 mg, 0.094
mmol, 1.0 equivalent) were dissolved in benzene (5 mL) then aqueous
2M Na.sub.2CO.sub.3 (1 mL) and Pd(PPh.sub.3).sub.4 (115.6 mg, 0.1
mmol, 0.2 equivalent) were added and the resulting suspension was
heated at reflux at 80.degree. C. for 16 hours. The reaction
mixture was diluted in ethyl acetate, washed with water and dried
over anhydrous sodium sulfate to afford
4-(5-chloro-2-isopropylamino-pyridin-4-yl)-1-(2,4,6-trimethyl-benz-
enesulfonyl)-1H-pyrrole-2-carboxylic acid methyl ester N-oxide
(R.sub.t(min) 6.859. MS FIA: 492.0 ES+) which was then treated with
a 2 M solution of PCl.sub.3 in dichloromethane (1 mL) at room
temperature. After 10 minutes, the solvent was removed under a
stream of nitrogen and the crude oil was dissolved in methanol (1
mL) and aqueous 1 M NaOH (1 mL). The resulting mixture was heated
at reflux for 16 hours then the crude solution was acidified using
aqueous 1 M HCl and the solvent was removed. The resulting
4-(5-chloro-2-isopropylamino-pyridin-4-yl)-1H-pyrrole-2-carboxylic
acid (R.sub.t(min) 3.527. MS FIA: 279.4 ES+; 278.2 Es-) was
suspended in DMF (3 mL) together with EDCI (36 mg, 0.19 mmol, 2
equivalents), HOBt (26 mg, 0.19 mmol, 2 equivalents),
(S)-3-chlorophenylglycinol HCl salt (59 mg, 0.28 mmol, 3
equivalents) and DIEA (0.12 mL, 0.75 mmol, 8 equivalents). The
resulting mixture was stirred at room temperature for 16 hours. The
reaction mixture was diluted in ethyl acetate, washed with water
and dried over sodium sulfate. After removing the solvent under
reduced pressure, the crude product was purified by reversed phase
HPLC (acetonitrile/water/TFA 1%) to afford the title compound as a
white solid (4.8 mg, 8.1%).
Example 3
Compound 1-3 was prepared as follows:
##STR00046##
2-Chloro-5-methyl-4-nitropyridine N-oxide: The title compound was
prepared in a manner substantially similar to that described by Z.
Talik, A. Puszko, Roczniki Chemii Ann. Soc. Chim. Polonorum 1976,
50, 2209, as follows. To a suspension of 2-chloro-5-methylpyridine
(10 g, 0.078 mol) in acetic anhydride (25 mL) was added hydrogen
peroxide 30% (25 mL) in small portions. This mixture was stirred at
room temperature for 24 hours and then heated at 60.degree. C. for
30 hours. After removing the excess acetic acid under reduced
pressure, the residue was added in small portions to concentrated
sulfuric acid (15 mL). The resulting solution was added to a
mixture of concentrated sulfuric acid (15 mL) and fuming nitric
acid (25 mL) and then heated at 100.degree. C. for 90 minutes. The
reaction mixture was poured onto ice, neutralized with solid
ammonium carbonate and finally with aqueos ammonia until a basic pH
was obtained and a precipitate formed. This precipitate was
collected by filtration to afford the title compound as a pale
yellow solid (9.4 g, 0.050 mol, R.sub.t(min) 3.272, FIA ES+ 188.9,
ES- 188.0).
2-((S)-1-Hydroxymethylpropylamino)-5-methyl-4-nitro-pyridine
N-oxide: 2-Chloro-5-methyl-4-nitropyridine N-oxide (200 mg, 1.06
mmol, 1.0 equivalent) was dissolved in ethanol (1.5 mL).
(S)-2-Aminobutanol (284 mg, 3.2 mmol, 3.0 equivalent) was then
added and the resulting mixture was refluxed for 16 hours. The
crude solution was then purified by reversed phase HPLC
(acetonitrile/water/TFA) to afford the title compound as a brown
solid (146 mg, 0.61 mmol, R.sub.t(min) 3.787; FIA ES+ 241.8, ES-
not observed).
Acetic acid 2-(4-bromo-5-methylpyrinin-2-ylamino)-butyl ester:
2-((S)-1-Hydroxymethylpropylamino)-5-methyl-4-nitro-pyridine
N-oxide (146 mg, 0.61 mmol) was dissolved in acetyl bromide (1.5
mL). The mixture was then heated at 90.degree. C. for 3 hours. The
acteyl bromide was evaporated off under a stream of nitrogen and
the crude material was dissolved in a solution of 2M PCl.sub.3 in
dichloromethane (2 mL). The resulting mixture was stirred at room
temperature for 1 hour and the reaction mixture poured into an
aqueous solution of saturated NaHCO.sub.3 and extracted with ethyl
acetate. The organic layer was washed with water, the solvent was
then dried over Na.sub.2SO.sub.4 and removed under reduced pressure
to afford the title compound as a brown oil (149 mg, (R.sub.t(min)
4.146; FIA ES+ 300.9, ES- not observed).
4-[2-(S)-(1-Acetoxymethylpropylamino)-5-methylpyridin-4-yl]-1-(2,4,6-trim-
e-thylbenzensulfonyl)-1H-pyrrole-2-carboxylic acid methyl ester: To
a solution of acetic acid
2-(4-bromo-5-methylpyrinin-2-ylamino)-butyl ester (149 mg, 0.5
mmol, 1.0 equivalent) and
4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1-(2,4,6-trimethylbenzen-
sulfonyl)-1H-pyrrole-2-carboxylic acid methyl ester (215 mg, 0.50
mmol, 1.0 equivalent) in benzene (5 mL) was added aqueous 2M
Na.sub.2CO.sub.3 (1 mL) and Pd(PPh.sub.3).sub.4 (115.6 mg, 0.1
mmol, 0.2 equivalent). After heating at reflux for 16 hours, the
mixture was poured in water and extracted with ethyl acetate. The
organic extract was dried over Na.sub.2SO.sub.4 and the solvent
removed under reduced pressure to afford the title compound
(R.sub.t(min) 6.684; FIA ES+ 528.3, ES- not observed) which was
carried into the next step.
4-[2-(S)-Hydroxymethylpropylamino)-5-methylpyridin-4-yl]-1H-pyrrole-2-car-
boylic acid [1-(S)-(3-chlorophenylgylcinol]amide (1-3): The crude
4-[2-(S)-(1-acetoxymethylpropylamino)-5-methylpyridin-4-yl]-1-(2,4,6-trim-
ethylbenzensulfonyl)-1H-pyrrole-2-carboxylic acid methyl ester was
dissolved in methanol (1.5 mL) and aquous 1M, NaOH (2 mL) and the
mixture heated at reflux for 16 hours. After acidifying with
aqueous 1M HCl (2.2 mL), the solvent was removed under reduced
pressure and the crude was then suspended in DMF (5 mL). After
adding EDCI (192 mg, 1.0 mmol), HOBt (135 mg, 1.0 mmol) and DIEA
(0.48 mL, 3.0 mmol), the mixture was stirred for 30 minutes at room
temperature before adding. (S)-3-chlorophenylglycinol HCl salt (312
mg, 1.5 mmol) and the reaction mixture was then stirred for 16
hours at room temperature. The reaction mixture was dissolved in
ethyl acetate, washed with water, the organic layer was dried over
Na.sub.2SO.sub.4 and the solvent was removed under reduced
pressure. The crude residue was purified by reversed phase HPLC
(acetonitrile/water/TFA) to afford the title compound as a
colorless solid (29.3 mg, 0.07 mmol, R.sub.t(min) 4.563; FIA ES+
443.1, ES- 441.5, ES+ 443.1, ES- 441.5; .sup.1HNMR (CD.sub.3OD)
.delta. 1.0 (t, 3H), 1.50 (m, 1H), 1.7 (m, 1H), 2.3 (s, 3H), 3.5
(m, 1H), 3.7 (m, 2H), 3.8 (m, 2H), 5.1 (t, 1H), 7.0 (s, 1H), 7.3
(m, 4H), 7.4 (two s, 2H), 7.6 (s, 1H).
Example 4
Characterization Data
Compounds of the present invention were prepared by methods
substantially similar to those described in the above Examples 1-3
and by methods known to one of ordinary skill in the art. The
characterization data for these compounds is summarized in Table 3
below and includes HPLC, MS, and .sup.1H NMR data. Unless specified
otherwise, the .sup.1H NMR data was obtained at 500 MHz and all
reported chemical shifts are ppm. Compound numbers correspond to
the compound numbers listed in Tables 1, 2, and 3. As used herein,
the term "R.sub.t" refers to the retention time, in minutes,
obtained for the designated compound using the HPLC method
described above. Where more than one analytic measurement was
obtained for any given compound, as described herein, only a single
exemplary measurement is provided.
TABLE-US-00003 TABLE 3 Characterization Data for Selected Compounds
of Formula I Cmpd # M + 1 R.sub.t .sup.1H NMR I-1 413.00 2.10
(CD.sub.3OD)1.3(d, 6H), 2.3(s, 3H), 3.7(m, 3H), 5.1(2, 1H), 6.9(s,
1H), 7.2(m, 4H), 7.4(bs, 2H), 7.6(s, 1H) I-2 379.20 2.00
(CD.sub.3OD)1.3(d, 6H), 2.3(s, 3H), 3.85(m, 3H), 5.15(t, 1H), 6.9
9s, 1H), 7.2(t, 1H), 7.3(m, 6H), 7.55(s, 1H) I-3 443.10 2.10
(CD.sub.3OD)1.0(t, 3H), 1.5(m, 1H), 1.7(m, 1H), 2.3(s, 3H), 3.5(m,
1H), 3,7(m, 2H), 3.8(m, 2H), 5.1(t, 1H), 7.0(s, 1H), 7.3(m, 4H),
7.4(two s, 2H), 7.6(s, 1H) I-4 393.30 2.10 (CD.sub.3OD)1.0(t, 3H),
1.3(d, 3H), 1.65(m, 2H), 2.35(s, 3H), 3.6(m, 1H), 3.8(m, 2H),
5.15(t, 1H), 6.95(s, 1H), 7.2(t, 1H), 7.3(2, 3H), 7.4(d, 1H),
7.45(s, 1H), 7.6(s, 1H) I-5 427.20 2.40 (CD.sub.3OD)1.0(t, 3H),
1.2(d, 3H), 1.6(m, 2H), 2.3(s, 3H), 3.75(m, 1H), 3.85(m, 2H),
5.15(t, 1H), 6.95(s, 1H), 7.3(m, 4H), 7.5(two s, 2H), 7.6(s, 1H)
I-6 427.10 2.30 (CD.sub.3OD)0.9(, 3H), 1.25(d, 3H), 1.55(m, 2H),
2.25(s, 3H), 3.7(m, 1H), 3.85(m, 2H), 5.15(t, 1H) I-7 427.10 2.40
(CD.sub.3OD)1.1(d, 6H), 1.9(m, 1H), 2.35(s, 3H), 3.15(d, 1H),
3.8(m, 2H), 5.2(m, 1H), 7.0(s, 1H), 7.3(m, 4H), 7.4(s, 1H), 7.6(s,
1H) I-8 411.10 2.20 (CD.sub.3OD)0.65(m, 2H), 0.95(m, 2H), 2.4(s,
3H), 2.65(m, 1H), 3.8(m, 2H), 5.15(t, 1H), 7.0(s, 1H), 7.2(t, 1H),
7.3(m, 3H), 7.4(s, 1H), 7.45(s, 1H), 7.75(s, 1H) I-9 433.70 2.30
(CD.sub.3OD)1.2(d, 6H), 3.8(m, 2H), 3.85(m, 1H), 5.1(t, 1H), 7.0(s,
1H), 7.2(m, 3H), 7.35(s, 1H), 7.4(7.65(s, 1H), 7.9(s, 1H) I-10
399.90 2.10 (CD.sub.3OD)1.2(d, 6H), 3.8(d, 2H), 3.9(m, 1H), 5.1(t,
1H), 6.6(s, 1H), 7.2(t, 1H), 7.3(m, 5H), 7.45(s, 1H), 7.9(s, 1H)
I-11 415.10 1.90 (CD.sub.3OD)1.25(d, 3H), 3.5(m, 1H), 3.7(m, 1H),
3.8(m, 2H), 3.9(m, 1H), 5.2(t, 1H), 7.2(t, 1H), 7.3(t, 2H), 7.35(m,
2H), 7.45(s, 1H), 7.65(s, 1H), 7.9(s, 1H) I-12 449.00 2.20
(CD.sub.3OD)1.2(d, 3H), 3.5(m, 1H), 3.7(m, 1H), 7.3(m, 3H), 7.4(s,
1H), 7.45(s, 1H), 7.7(s, 1H), 7.9(s, 1H) I-13 462.90 2.20
(CD.sub.3OD)1.0(t, 3H), 1.6(m, 1H), 1.7(m, 1H), 3.55(m, 1H), 3.7(m,
2H), 3.8(m, 2H), 5.15(2, 1H), 7.1(s, 1H), 7.2(m, 3H), 7.4(s, 1H),
7.5(s, 1H), 7.7(s, 1H), 7.9(s, 1H) I-14 429.00 2.00
(CD.sub.3OD)1.0(t, 3H), 1.55(m, 1H), 1,75(m, 1H), 3.5(m, 1H),
3.7(m, 2H), 3.8(m, 2H), 5.1(t, 1H), 7.1(s, 1H), 7.2(d, 1H), 7.3(t,
2H), 7.35(m, 2H), 7.4(s, 1H), 7.7(s, 1H), 7.9(s, 1H) I-15 447.10
2.50 (CD.sub.3OD)1.0(t, 2H), 1.3(d, 3H), 1.7(m, 2H), 3.7(m, 1H),
3.85(m, 2H), 5.15(2, 1H), 7.1(s, 1H), 7.25(m, 3H). 7.4(s, 1H),
7.5(s, 1H), 7.7(s, 1H) I-16 477.00 2.40 (CD.sub.3OD)1.0(t, 314),
1.6(m, 1H), 1.8(m, 1H), 3.6(m, 1H), 3.7(m, 2H), 3.85(m, 2H), 3.9(s,
3H), 5.1(t, 1H), 7.1(s, 1H), 7.25(m, 1H), 7.3(m, 2H), 7.4(2s, 2H),
7.7(s, 1H), 7.95(s, 1H) I-17 433.00 2.30 (CD.sub.3OD): 8.0(s, 1H),
7.7(s, 1H), 7.25-7.5(m, 6H), 5.15(m, 1H), 3.8-3.95(m, 3H), 1.3(d,
6H) I-18 449.00 2.36 (CD.sub.3OD)7.96(s, 1H); 7.7(s, 1H); 7.48(s,
1H); 7.42(s, 1H); 7.32(s, 2H); 7.24(s, 2H); 7.2(s, 1H); 5.15(t,
1H); 3.8-4.0(m. 5H); 3.72(m, 1H); 3.57(m, 1H); 1.3(s, 3H)
Example 5
Synthesis of Prodrugs
Prodrugs of formula II are prepared from the hydroxyl compounds of
formula I by a variety of methods known to one of ordinary skill in
the art. These methods include, but are not limited to, acylation
by a desired carboxylic acid or phosphate formation. When the
hydroxyl moiety of formula 1 is acylated by a desired amino acid,
the amino moiety of the amino acid may be optionally protected by a
suitable amino protecting group as described herein supra.
The preparation of the L-valine prodrug of compound 1-9 is
described in detail below.
##STR00047##
(2S)-(S)-2-(4-(5-chloro-2-(isopropylamino)pyridine-4-yl)-1H-pyrrole-2-car-
boxamido)-2-(3-chlorophenyl)ethyl-2-amino-2-methylbutanoate (II-1):
To a solution of compound 1-9 (1 g, 2.3 mmol, 1.0 equivalent) in
dichloromethane (50 mL) were added DIEA (1.1 mL, 6.9 mmol, 3.0
equivalent) and N--BOC-L-Valine (1.2 g, 5.52 mmol, 2.4
equivalents). PyBOP (2.9 g, 5.75 mmol, 2.5 equivalents.) was then
added slowly and the resulting mixture was stirred at room
temperature for 48 hours. The mixture was then washed with water
and dried over sodium sulfate. The crude solid was adsorbed on
silica and then purified by flash chromatography eluting with
mixtures of hexane/ethyl acetate (from 90:10 to 50:50), yielding
the Boc-protected compound as a white solid (786 mg). This
intermediate (761 mg, 1.2 mmol) was dissolved in dioxane (1 mL) and
treated with a solution of 4 M HCL in dioxane. The resulting
mixture was stirred for 16 hours at room temperature. The solvent
was then removed and the 2.times.HCl salt of the title compound was
obtained as a white solid (571 mg). HPLC R.sub.t: 4.56 minutes. FIA
MS: 531.9 ES+; 529.8 ES-. LC/MS: R.sub.t: 2.07 minutes; 532.0 ES+;
530.1 ES-. .sup.1HNMR (CD.sub.3OD) .delta. 0.9 (dd, 6H), 1.35 (d,
6H), 2.2 (m, 1H), 3.9 (m, 2H), 4.7 (m, 2H), 5.6 (m, 1H), 7.1 (s,
1H), 7.3 (d, 1H), 7.35 (t, 1H), 7.4 (d, 1H), 7.5 (s, 1H), 7.6 (s,
1H), 7.75 (s, 1H), 7.95 (s, 1H).
The preparation of a phosphate prodrug of compound 1-9 is described
in detail below.
##STR00048##
di-tert-Butyl
4-(5-chloro-2-(isopropylamino)pyridine-4-yl)-N-((S)-1-(3-chlorophenyl)-2--
hydroxyethyl)-1H-pyrrole-2-carboxamide phosphate: Compound 1-9 (1
g, 2.3 mmol, 1.0 equivalent) and tetrazole (241 mg, 3.45 mmol, 1.5
equivalents) were dissolved in dichloromethane (5 mL) and
acetonitrile (5 mL) under nitrogen at room temperature.
Di-tert-butyl diisopropyl phosphoamidite (1.1 mL, 3.45 mmol, 1.5
equivalents) was added dropwise and the resulting mixture was
stirred for 16 hours. The reaction mixture was then cooled on an
ice bath, treated with a solution of 6 M tert-butylhydroxyperoxide
(3 mL) and stirred for 20 minutes. The clear solution was diluted
in dichloromethane and small amount of methanol, washed with
Na.sub.2S.sub.2O.sub.3, water and dried over sodium sulfate. The
crude oil was adsorbed on silica gel and was first purified by
flash chromatography eluting with mixtures of hexane/acetone (from
90:10 to 60:40) and then by reversed phase HPLC
(acetonitrile/water/1% TFA), yielding the di-t-butyl ether
intermediate as a white solid (336 mg). HPLC R.sub.t: 6.53 minutes,
MS FIA: 625.0 ES+; 623.1 ES-.
4-(5-Chloro-2-(isopropylamino)pyridine-4-yl)-N-((S)-1-(3-chlorophenyl)-2--
hydroxyethyl)-1H-pyrrole-2-carboxamide phosphate (II-2): The
t-butyl phosphate intermediate (336 mg, 0.54 mmol) was suspended in
dioxane (5 mL) and a solution of 4 M HCL in dioxane was added. The
resulting mixture was stirred at room temperature for 1 hour. The
solvent was then removed and the free phosphate was then dissolved
in a solution of DMSO (15 mL), methanol (50 mL) and water (25 mL)
and treated with a solution of 2 M Na.sub.2CO.sub.3 (0.25 mL).
Methanol was removed under reduced pressure and the aqueous/DMSO
mixture was remove using to a liophilizer to afford the title
compound as an off white solid (187 mg). HPLC R.sub.t: 4.24
minutes. FIA MS: 512.9 ES+; 510.9 ES-. LC/MS: R.sub.t 2.39 minutes;
512.9 ES+; 510.9 ES-. .sup.1HNMR (CD.sub.3OD) .delta. 1.2 (d, 6H),
3.9 (m, 1H), 4.1 (dm, 2H), 5.1 (m, 1H), 6.7 (s, 1H), 7.2 (d, 1H),
7.25 (t, 1H), 7.3 (m, 2H), 7.45 (s, 1H), 7.55 (s, 1H), 7.9 (s,
1H).
Example 6
ERK2 Inhibition Assay
Compounds were assayed for the inhibition of ERK2 by a
spectrophotometric coupled-enzyme assay (Fox et al (1998) Protein
Sci 7, 2249). In this assay, a fixed concentration of activated
ERK2 (10 nM) was incubated with various concentrations of the
compound in DMSO (2.5%) for 10 minutes at 30.degree. C. in 0.1 M
HEPES buffer, pH 7.5, containing 10 mM MgCl.sub.2, 2.5 mM
phosphoenolpyruvate, 200 .mu.M NADH, 150 .mu.g/mL pyruvate kinase,
50 .mu.g/mL lactate dehydrogenase, and 200 .mu.M erktide peptide.
The reaction was initiated by the addition of 65 .mu.M ATP. The
rate of decrease of absorbance at 340 nM was monitored. The
IC.sub.50 was evaluated from the rate data as a function of
inhibitor concentration.
Compounds of the present invention we found to be inhibitors of
ERK2 protein kinase. In certain embodiments, compounds were found
to inhibit ERK2 kinase at <0.1 .mu.M. In other embodiments,
compounds were found to inhibit ERK2 kinase at <0.01 .mu.M.
Example 7
ERK1 Inhibition Assay
Compounds are assayed for the inhibition of ERK1 by a
spectrophotometric coupled-enzyme assay (Fox et al (1998) Protein
Sci 7, 2249). In this assay, a fixed concentration of activated
ERK1 (20 nM) is incubated with various concentrations of the
compound in DMSO (2.0%) for 10 minutes at 30.degree. C. in 0.1 M
HEPES buffer, pH 7.6, containing 10 mM MgCl.sub.2, 2.5 mM
phosphoenolpyruvate, 200 .mu.M NADH, 30 .mu.g/mL pyruvate kinase,
10 .mu.g/mL lactate dehydrogenase, and 150 .mu.M erktide peptide.
The reaction is initiated by the addition of 140 .mu.M ATP (20
.mu.L). The rate of decrease of absorbance at 340 nM is monitored.
The K.sub.i is evaluated from the rate data as a function of
inhibitor concentration.
While we have described a number of embodiments of this invention,
it is apparent that our basic examples may be altered to provide
other embodiments that utilize the compounds and methods of this
invention. Therefore, it will be appreciated that the scope of this
invention is to be defined by the appended claims rather than by
the specific embodiments that have been represented by way of
example.
* * * * *
References