U.S. patent application number 13/018337 was filed with the patent office on 2011-06-16 for anti-inflammatory pyrazolopyrimndines.
This patent application is currently assigned to The Regents of the University of California. Invention is credited to Heinz W. Gschwend, Eric J. Kunkel, Nikolai Sepetov, Kevan M. Shokat, Chao Zhang.
Application Number | 20110144134 13/018337 |
Document ID | / |
Family ID | 36218790 |
Filed Date | 2011-06-16 |
United States Patent
Application |
20110144134 |
Kind Code |
A1 |
Shokat; Kevan M. ; et
al. |
June 16, 2011 |
Anti-Inflammatory Pyrazolopyrimndines
Abstract
The present invention provides anti-inflammatory compounds
useful in the treatment of diseases and conditions in which
inflammation is involved in disease progression or the
manifestation of symptoms of the disease or condition.
Inventors: |
Shokat; Kevan M.; (San
Francisco, CA) ; Sepetov; Nikolai; (Los Gatos,
CA) ; Zhang; Chao; (San Francisco, CA) ;
Gschwend; Heinz W.; (Santa Rosa, CA) ; Kunkel; Eric
J.; (San Mateo, CA) |
Assignee: |
The Regents of the University of
California
Oakland
CA
|
Family ID: |
36218790 |
Appl. No.: |
13/018337 |
Filed: |
January 31, 2011 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11719722 |
Dec 5, 2008 |
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PCT/US05/42524 |
Nov 21, 2005 |
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13018337 |
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60629639 |
Nov 19, 2004 |
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Current U.S.
Class: |
514/262.1 ;
544/262 |
Current CPC
Class: |
A61P 1/00 20180101; A61P
1/02 20180101; A61P 29/00 20180101; A61P 31/04 20180101; A61P 9/04
20180101; A61P 19/02 20180101; A61P 1/04 20180101; A61P 25/00
20180101; A61P 25/24 20180101; A61P 25/28 20180101; A61P 27/02
20180101; A61P 3/10 20180101; A61P 43/00 20180101; A61P 21/04
20180101; A61P 17/06 20180101; A61P 11/00 20180101; A61P 15/00
20180101; A61P 9/00 20180101; C07D 487/04 20130101; A61P 3/04
20180101; A61P 25/16 20180101; A61P 7/04 20180101; A61P 25/06
20180101; A61P 37/02 20180101; A61P 17/00 20180101; A61P 33/02
20180101; A61P 19/04 20180101; A61P 31/00 20180101; A61P 7/06
20180101; A61P 9/14 20180101; A61P 9/10 20180101; A61P 11/06
20180101; A61P 35/00 20180101; A61P 25/22 20180101; A61P 17/02
20180101; A61P 19/10 20180101; A61P 25/14 20180101 |
Class at
Publication: |
514/262.1 ;
544/262 |
International
Class: |
A61K 31/519 20060101
A61K031/519; C07D 487/04 20060101 C07D487/04; A61P 29/00 20060101
A61P029/00; A61P 31/00 20060101 A61P031/00; A61P 9/00 20060101
A61P009/00; A61P 35/00 20060101 A61P035/00; A61P 3/04 20060101
A61P003/04; A61P 3/10 20060101 A61P003/10; A61P 1/00 20060101
A61P001/00; A61P 17/06 20060101 A61P017/06; A61P 19/02 20060101
A61P019/02; A61P 11/06 20060101 A61P011/06; A61P 25/28 20060101
A61P025/28 |
Goverment Interests
STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED
RESEARCH AND DEVELOPMENT
[0002] This invention was made with government support under grant
AI 044009 awarded by the National Institutes of Health. The
Government has certain rights in the invention.
Claims
1. A compound having the formula: ##STR00101## wherein R.sup.1 is
substituted or unsubstituted heteroaryl; R.sup.2, R.sup.3, and
R.sup.4 are independently hydrogen, substituted or unsubstituted
alkyl, substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, or substituted
or unsubstituted heteroaryl; and L.sup.1 is a bond, substituted or
unsubstituted alkylene, substituted or unsubstituted cycloalkylene
or substituted or unsubstituted heteroalkylene.
2. A compound according to claim 1, wherein R.sup.2 is
R.sup.10-substituted C.sub.1-C.sub.20 alkyl, 2 to 20 membered
substituted or unsubstituted heteroalkyl, R.sup.11-substituted
C.sub.3-C.sub.8 cycloalkyl, R.sup.11-substituted C.sub.3-C.sub.8
heterocycloalkyl, R.sup.12-substituted heteroaryl, or
R.sup.13-substituted aryl, wherein R.sup.10 is oxo, --OH, halogen,
--CF.sub.3, --NH.sub.2, 2 to 20 membered substituted or
unsubstituted heteroalkyl, 3 to 7 membered substituted or
unsubstituted heterocycloalkyl, substituted or unsubstituted
heteroaryl, or R.sup.13-substituted aryl, R.sup.11 is oxo, --OH,
halogen, --CF.sub.3, --NH.sub.2, substituted or unsubstituted
alkyl, substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, or substituted
or unsubstituted heteroaryl, R.sup.12 is --OH, halogen, --CF.sub.3,
--NH.sub.2, substituted or unsubstituted alkyl, substituted or
unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted heterocycloalkyl, substituted or
unsubstituted aryl, or substituted or unsubstituted heteroaryl, and
R.sup.13 is --OH; --NH.sub.2, substituted or unsubstituted
cycloalkyl, substituted or unsubstituted heterocycloalkyl,
substituted or unsubstituted heteroaryl, or substituted or
unsubstituted aryl.
3. A compound according to claim 1, wherein R.sup.1 is
R.sup.15-substituted heteroaryl, and further wherein R.sup.15 is
--OH, halogen, --CF.sub.3, --NH.sub.2, substituted or unsubstituted
alkyl, substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, or substituted
or unsubstituted heteroaryl.
4. A compound according to claim 1, wherein R.sup.1 is substituted
or unsubstituted fused-ring heteroaryl.
5. A compound according to claim 1, R.sup.3 and R.sup.4 are
hydrogen
6. A method for treating a disorder, said method comprising
administering to a subject in need thereof a therapeutically
effective amount of a compound according to claim 1.
7. The method of claim 6, wherein R.sup.1 is R.sup.15-substituted
heteroaryl, and further wherein R.sup.15 is --OH, halogen,
--CF.sub.3, --NH.sub.2, substituted or unsubstituted alkyl,
substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, or substituted
or unsubstituted heteroaryl.
8. The method of claim 6, wherein R.sup.1 is substituted or
unsubstituted fused-ring heteroaryl.
9. The method of claim 6, wherein R.sup.3 and R.sup.4 are
hydrogen.
10. The method of claim 6, wherein said disorder is an inflammatory
process that occurs in response to infection, trauma, autoimmune
disease, cardiovascular disease, neoplasia, hyperplasia, addiction,
infection, obesity, cellular degeneration, apoptosis, or
senescence, or differentiation.
11. The method of claim 6, wherein said disorder is selected from
the group consisting of vasculitis, multiple sclerosis, diabetes,
inflammatory bowel disease, psoriasis, rheumatoid arthritis,
Crohn's disease, ulcerative colitis, asthma, stroke,
atherosclerosis, and lupus.
12. The method of claim 6, wherein said disorder is selected from
the group consisting of headaches, bronchitis, menstrual cramps,
tendonitis, gastritis, vascular diseases, uveitis, Sjogren's
disease, sclerodoma, nephrotic syndrome, swelling occurring after
injury, myocardial ischemia, fever, common cold, dysmenorrhea,
emphysema, acute respiratory distress syndrome, chronic obstructive
pulmonary disease, Alzheimer's disease, organ transplant toxicity,
cachexia, allergic reactions, allergic contact hypersensitivity,
cancer, tissue ulceration, peptic ulcers, regional enteritis,
ulcerative colitis, diverticulitis, recurrent gastrointestinal
lesion, gastrointestinal bleeding, coagulation, synovitis, gout,
ankylosing spondylitis, restenosis, periodontal disease,
epidermolysis bullosa, osteoporosis, loosening of artificial joint
implants, aortic aneurysm, periarteritis nodosa, congestive heart
failure, myocardial infarction, cerebral ischemia, head trauma,
spinal cord injury, neuralgia, neuro-degenerative disorders,
autoimmune disorders, Huntington's disease, Parkinson's disease,
migraine, depression, peripheral neuropathy, pain, gingivitis,
cerebral amyloid angiopathy, nootropic or cognition enhancement,
amyotrophic lateral sclerosis, multiple sclerosis, ocular
angiogenesis, corneal injury, macular degeneration, conjunctivitis,
abnormal wound healing, muscle or joint sprains or strains,
tendonitis, skin disorders, myasthenia gravis, polymyositis,
myositis, bursitis, burns, diabetes, tumor invasion, tumor growth,
tumor metastasis, corneal scarring, scleritis, immunodeficiency
diseases, sepsis, premature labor, hypoprothrombinemia, hemophilia,
thyroiditis, sarcoidosis, Behcet's syndrome, hypersensitivity,
kidney disease, Rickettsial infections, Protozoan diseases,
reproductive disorders, and septic shock.
Description
CROSS-REFERENCES TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 11/719,722, filed Dec. 5, 2008, which claims
benefit of PCT/US2005/42524 filed Nov. 21, 2005, which claims the
benefit of U.S. Provisional Patent Application No. 60/629,639,
filed Nov. 19, 2004, each of which is incorporated herein by
reference in its entirety for all purposes.
BACKGROUND OF THE INVENTION
[0003] Protein kinases are involved in a wide variety of cellular
processes, such as growth factor response, cytokine response,
immune response, stress response, and cell cycle regulation, to
specify only a few such processes. Because their improper
regulation is believed to cause various diseases including cancer
and inflammation, protein kinases are important drug targets for
the treatment of these diseases.
[0004] Inflammatory conditions, particularly chronic inflammatory
diseases, are of particular interest to developers of new
pharmaceutical products, because such conditions and diseases are
widespread, and improved therapies for them are needed. These
diseases are caused by action of the immune system, including the
inappropriate activation of T cells, expression of regulatory
cytokines and chemokines, loss of immune tolerance, and the like.
Modulation of the immune response varies with the specific factors
produced and the receptors present on the responding cell. Among
these diseases are autoimmune and/or chronic inflammatory diseases,
which include multiple sclerosis and inflammatory bowel diseases
("IBD," including ulcerative colitis and Crohn's disease), colitis,
diseases of the joints, such as rheumatoid arthritis, diseases
involving the destruction or improper alteration of nucleic acids,
as observed with systemic lupus erythematosus and other diseases
such as psoriasis, insulin dependent diabetes mellitus (IDDM),
Sjogren's disease, myasthenia gravis, thyroid disease, Alzheimer's
disease, uveitis, and cardiovascular disease.
[0005] Thus, there is a need in the art for effective
anti-inflammatory compounds useful in the treatment of disease. The
present invention addresses these and other needs.
BRIEF SUMMARY OF THE INVENTION
[0006] The present invention provides anti-inflammatory compounds
useful in the treatment of diseases and conditions in which
inflammation is involved in disease progression or the
manifestation of symptoms of the disease or condition.
[0007] In one aspect, the present invention provides a compound
represented by the following structural formula:
##STR00001##
[0008] In Formula (I) above, R.sup.1 is selected from substituted
or unsubstituted alkyl, substituted or unsubstituted heteroalkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocycloalkyl, substituted or unsubstituted aryl,
or substituted or unsubstituted heteroaryl.
[0009] R.sup.2, R.sup.3, and R.sup.4 are, independently, hydrogen,
substituted or unsubstituted alkyl, substituted or unsubstituted
heteroalkyl, substituted or unsubstituted cycloalkyl, substituted
or unsubstituted heterocycloalkyl, substituted or unsubstituted
aryl, or substituted or unsubstituted heteroaryl. R.sup.3 and
R.sup.4 are optionally joined with the nitrogen to which they are
attached to form a substituted or unsubstituted heterocycloalkyl or
substituted or unsubstituted heteroaryl.
[0010] L.sup.1 is a bond, substituted or unsubstituted alkylene,
substituted or unsubstituted cycloalkylene, or substituted or
unsubstituted heteroalkylene.
[0011] In another aspect, the present invention provides a method
for treating or preventing a disorder characterized by abnormal
inflammation, said method including administering to a subject a
therapeutically effective amount of a compound of the present
invention.
[0012] In another aspect, the present invention provides
pharmaceutical formulations of a compound of the present invention,
and methods for using such compounds and their pharmaceutical
formulations in the treatment of inflammatory diseases and
conditions.
[0013] In another aspect, the present invention provides methods
for making the compounds and pharmaceutical formulations of the
invention.
[0014] In another aspect, the present invention provides compounds
and pharmaceutical formulations for use in the treatment of an
inflammatory disease or condition.
[0015] In certain embodiments, the compounds of the invention are T
cell activation inhibitors. In another embodiment, the compounds of
the invention are monocyte activation inhibitors. In another
embodiment, the compounds of the invention are gamma interferon
signaling inhibitors.
BRIEF DESCRIPTION OF THE DRAWINGS
[0016] FIG. 1 illustrates four exemplary BioMAP.TM. (BioSeek, Inc.,
Burlingame, Calif.) inflammation model systems useful in detecting
and discriminating modifiers of multiple targets and pathways,
where panel (A) sets forth multiple therapeutically relevant
targets and pathways that are detected and discriminated in BioMAP
models, including a large number of kinases, and panel (B) provides
details on the four exemplary model systems.
[0017] FIG. 2 illustrates the decrease in the number of recruited
leukocytes in a mouse model of peritoneal inflammation resulting
from the administration of an anti-inflammatory pyrazolopyrimidine
of the present invention.
[0018] FIG. 3 illustrates the decrease in macrophage and neutrophil
recruitment in a mouse model of peritoneal inflammation resulting
from the administration of an anti-inflammatory pyrazolopyrimidine
of the present invention.
DETAILED DESCRIPTION OF THE INVENTION
Definitions
[0019] Abbreviations used herein have their conventional meaning
within the chemical and biological arts.
[0020] Where substituent groups are specified by their conventional
chemical formulae, written from left to right, they equally
encompass the chemically identical substituents that would result
from writing the structure from right to left, e.g., --CH.sub.2O--
is equivalent to --OCH.sub.2--.
[0021] The term "alkyl," by itself or as part of another
substituent, means, unless otherwise stated, a straight (i.e.
unbranched) or branched chain, or cyclic hydrocarbon radical, or
combination thereof, which may be fully saturated, or mono- or
polyunsaturated, and can include di- and multivalent radicals,
having the number of carbon atoms designated (i.e. C.sub.1-C.sub.10
means one to ten carbons). Examples of saturated hydrocarbon
radicals include, but are not limited to, groups such as methyl,
ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl,
cyclohexyl, (cyclohexyl)methyl, cyclopropylmethyl, and homologs and
isomers of, for example, n-pentyl, n-hexyl, n-heptyl, n-octyl, and
the like. An unsaturated alkyl group is one having one or more
double bonds or triple bonds or both. Examples of unsaturated alkyl
groups include, but are not limited to, vinyl, 2-propenyl, crotyl,
2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl,
3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, and 3-butynyl, as
well as the higher homologs and isomers. Alkyl groups that are
limited to hydrocarbon groups are termed "homoalkyl".
[0022] The term "alkylene," by itself or as part of another
substituent, means a divalent radical derived from an alkyl, as
exemplified, but not limited, by
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2--. Typically, an alkyl (or
alkylene) group will have from 1 to 24 carbon atoms, with those
groups having 10 or fewer carbon atoms being preferred in the
present invention. A "lower alkyl" or "lower alkylene" is a shorter
chain alkyl or alkylene group, generally having eight or fewer
carbon atoms.
[0023] The term "heteroalkyl," by itself or in combination with
another term, means, unless otherwise stated, a straight or
branched chain, or cyclic hydrocarbon radical, or combinations
thereof, consisting of at least one carbon atom and at least one
heteroatom selected from the group consisting of O, N, P, Si and S,
and wherein the nitrogen and sulfur atoms may optionally be
oxidized and the nitrogen heteroatom may optionally be quaternized.
The heteroatom(s) O, N, P, S and Si may be placed at any interior
position of the heteroalkyl group or at the position at which alkyl
group is attached to the remainder of the molecule. Examples
include, but are not limited to, --CH.sub.2--CH.sub.2--O--CH.sub.3,
--CH.sub.2--CH.sub.2--NH--CH.sub.3,
--CH.sub.2--CH.sub.2--N(CH.sub.3)--CH.sub.3,
--CH.sub.2--S--CH.sub.2--CH.sub.3, --CH.sub.2--CH.sub.2,
--S(O)--CH.sub.3, --CH.sub.2--CH.sub.2--S(O).sub.2--CH.sub.3,
--CH.dbd.CH--O--CH.sub.3, --Si(CH.sub.3).sub.3,
--CH.sub.2--CH.dbd.N--OCH.sub.3,
--CH.dbd.CH--N(CH.sub.3)--CH.sub.3, O--CH.sub.3,
--O--CH.sub.2--CH.sub.3, and --CN. Up to two heteroatoms may be
consecutive, such as, for example, --CH.sub.2--NH--OCH.sub.3 and
--CH.sub.2--O--Si(CH.sub.3).sub.3. Similarly, the term
"heteroalkylene," by itself or as part of another substituent,
means a divalent radical derived from heteroalkyl, as exemplified,
but not limited to, --CH.sub.2--CH.sub.2--S--CH.sub.2--CH.sub.2--
and --CH.sub.2--S--CH.sub.2--CH.sub.2--NH--CH.sub.2--. For
heteroalkylene groups, heteroatoms can also occupy either or both
of the chain termini (e.g., alkyleneoxy, alkylenedioxy,
alkyleneamino, alkylenediamino, and the like). Still further, for
alkylene and heteroalkylene linking groups, no orientation of the
linking group is implied by the direction in which the formula of
the linking group is written. For example, the formula
--C(O).sub.2R'-- represents both --C(O).sub.2R'-- and
--R'C(O).sub.2--. As described above, heteroalkyl groups, as used
herein, include those groups that are attached to the remainder of
the molecule through a heteroatom, such as --C(O)R', --C(O)NR',
--NR'R'', --OR', --SR', and/or --SO.sub.2R'. Where "heteroalkyl" is
recited, followed by recitations of specific heteroalkyl groups,
such as --NR'R'' or the like, it will be understood that the terms
heteroalkyl and --NR'R'' are not redundant or mutually exclusive.
Rather, the specific heteroalkyl groups are recited to add clarity.
Thus, the term "heteroalkyl" should not be interpreted as used
herein as excluding specific heteroalkyl groups, such as --NR'R''
or the like.
[0024] The term "alkoxy" refers to an alkyl attached to the
remainder of the molecule via an oxygen heteroatom. The alkyl
portion of the alkoxy group may be any appropriate length. In some
embodiments, the alkyl portion of the alkoxy group is a
C.sub.1-C.sub.20 alkyl (i.e. C.sub.1-C.sub.20 alkoxy). In some
embodiments, the alkyl portion of the alkoxy group is a
C.sub.1-C.sub.10 alkyl (i.e. C.sub.1-C.sub.10 alkoxy). In some
embodiments, the alkyl portion of the alkoxy group is a
C.sub.1-C.sub.10 alkyl (i.e. C.sub.1-C.sub.10 alkoxy).
[0025] An "alkylesteryl," as used herein, refers to a moiety having
the formula R'--C(O)O--R'', wherein R' is an alkylene moiety and
R'' is an alkyl moiety.
[0026] The terms "cycloalkyl" and "heterocycloalkyl", by themselves
or in combination with other terms, represent, unless otherwise
stated, cyclic versions of "alkyl" and "heteroalkyl", respectively.
Additionally, for heterocycloalkyl, a heteroatom can occupy the
position at which the heterocycle is attached to the remainder of
the molecule. Examples of cycloalkyl include, but are not limited
to, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl,
cycloheptyl, and the like. Examples of heterocycloalkyl include,
but are not limited to, 1-(1,2,5,6-tetrahydropyridyl),
1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl,
3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl,
tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1-piperazinyl,
2-piperazinyl, and the like. The terms "cycloalkylene" and
"heterocycloalkylene" refer to the divalent derivatives of
cycloalkyl and heterocycloalkyl, respectively.
[0027] The terms "halo" or "halogen," by themselves or as part of
another substituent, mean, unless otherwise stated, a fluorine,
chlorine, bromine, or iodine atom. Additionally, terms such as
"haloalkyl," are meant to include monohaloalkyl and polyhaloalkyl.
For example, the term "halo(C.sub.1-C.sub.4)alkyl" is mean to
include, but not be limited to, trifluoromethyl,
2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the
like.
[0028] The term "aryl" means, unless otherwise stated, a
polyunsaturated, aromatic, hydrocarbon substituent which can be a
single ring or multiple rings (preferably from 1 to 3 rings) which
are fused together or linked covalently. The term "heteroaryl"
refers to aryl groups (or rings) that contain from one to four
heteroatoms selected from N, O, and S, wherein the nitrogen and
sulfur atoms are optionally oxidized, and the nitrogen atom(s) are
optionally quaternized. A heteroaryl group can be attached to the
remainder of the molecule through a carbon or heteroatom.
Non-limiting examples of aryl and heteroaryl groups include phenyl,
1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl,
3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl,
2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl,
3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl,
5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl,
3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl,
purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl,
2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl.
Substituents for each of above noted aryl and heteroaryl ring
systems are selected from the group of acceptable substituents
described below. The terms "arylene" and "heteroarylene" refer to
the divalent derivatives of aryl and heteroaryl, respectively.
[0029] For brevity, the term "aryl" when used in combination with
other terms (e.g., aryloxy, arylthioxy, arylalkyl) includes both
aryl and heteroaryl rings as defined above in combination with
another moiety. Thus, the term "arylalkyl" is meant to include
those radicals in which an aryl group is attached to an alkyl group
(e.g., benzyl, phenethyl, pyridylmethyl and the like) including
those alkyl groups in which a carbon atom (e.g., a methylene group)
has been replaced by, for example, an oxygen atom (e.g.,
phenoxymethyl, 2-pyridyloxymethyl, 3-(1-naphthyloxy)propyl, and the
like). However, the term "haloaryl," as used herein is meant to
cover only aryls substituted with one or more halogens.
[0030] The term "oxo" as used herein means an oxygen that is double
bonded to a carbon atom.
[0031] Each of above terms (e.g., "alkyl," "heteroalkyl,"
"cycloalkyl, and "heterocycloalkyl", "aryl," "heteroaryl" as well
as their divalent radical derivatives) are meant to include both
substituted and unsubstituted forms of the indicated radical.
Preferred substituents for each type of radical are provided
below.
[0032] Substituents for alkyl, heteroalkyl, cycloalkyl,
heterocycloalkyl monovalent and divalent derivative radicals
(including those groups often referred to as alkylene, alkenyl,
heteroalkylene, heteroalkenyl, alkynyl, cycloalkyl,
heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl) can be one
or more of a variety of groups selected from, but not limited to:
--OR', .dbd.O, .dbd.NR', .dbd.N--OR', --NR'R'', --SR', -halogen,
--SiR'R''R''', --OC(O)R', --CO.sub.2R', --CONR'R'', --OC(O)NR'R'',
--NR''C(O)R', --NR'--C(O)NR''R''', --NR''C(O).sub.2R',
--NR--C(NR'R''R''').dbd.NR'''', --NR--C(NR'R'').dbd.NR''',
--S(O)R', --S(O).sub.2R', --S(O).sub.2NR'R'', --NRSO.sub.2R', --CN
and --NO.sub.2 in a number ranging from zero to (2m'+1), where m'
is the total number of carbon atoms in such radical. R', R'', R'''
and R'''' each independently refer to hydrogen, substituted or
unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted heterocycloalkyl, substituted or
unsubstituted aryl (e.g., aryl substituted with 1-3 halogens),
substituted or unsubstituted alkyl, alkoxy or thioalkoxy groups, or
arylalkyl groups. When a compound of the invention includes more
than one R group, for example, each of the R groups is
independently selected as are each R', R'', R''' and R'''' groups
when more than one of these groups is present. When R' and R'' are
attached to the same nitrogen atom, they can be combined with the
nitrogen atom to form a 4-, 5-, 6-, or 7-membered ring. For
example, --NR/R'' is meant to include, but not be limited to,
1-pyrrolidinyl and 4-morpholinyl. From the above discussion of
substituents, one of skill in art will understand that the term
"alkyl" is meant to include groups in which carbon atoms are bound
to groups other than hydrogen groups, such as haloalkyl (e.g.,
--CF.sub.3 and --CH.sub.2CF.sub.3) and acyl (e.g., --C(O)CH.sub.3,
--C(O)CF.sub.3, --C(O)CH.sub.2OCH.sub.3), and the like.
[0033] Similar to the substituents described for alkyl radicals
above, exemplary substituents for aryl and heteroaryl groups (as
well as their divalent derivatives) are varied and are selected
from, for example and without limitation: --OR', --NR'R'', --SR',
-halogen, --SiR'R''R''', --OC(O)R', --C(O)R', --CONR'R'',
--OC(O)NR'R'', --NR''C(O)R', --NR'--C(O)NR''R''',
--NR''C(O).sub.2R', --NR--C(NR'R''R''').dbd.NR'',
--NR--C(NR'R'').dbd.NR''', --S(O)R', --S(O).sub.2R',
--S(O).sub.2NR'R'', --NRSO.sub.2R', --CN and --NO.sub.2, --R',
--N.sub.3, --CH(Ph).sub.2, fluoro(C.sub.1-C.sub.4)alkoxy, and
fluoro(C.sub.1-C.sub.4)alkyl, in a number, when present, ranging
from one to the total number of open valences on aromatic ring
system; and where R', R'', R''' and R'''' are independently
selected from hydrogen, substituted or unsubstituted alkyl,
substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl and substituted
or unsubstituted heteroaryl. When a compound of the invention
includes more than one R group, for example, each of the R groups
is independently selected as are each R', R'', R''' and R''''
groups when more than one of these groups is present.
[0034] Two of the substituents on adjacent atoms of an aryl or
heteroaryl ring may optionally form a ring of the formula
--T--C(O)--(CRR').sub.q--U--, wherein T and U are independently
--NR--, --O--, --CRR'-- or a single bond, and q is an integer of
from 0 to 3. Alternatively, two of the substituents on adjacent
atoms of aryl or heteroaryl ring may optionally be replaced with a
substituent of the formula -A-(CH.sub.2).sub.r--B--, wherein A and
B are independently --CRR'--, --O--, --NR--, --S--, --S(O).sub.2--,
--S(O).sub.2NR'-- or a single bond, and r is an integer of from 1
to 4. One of the single bonds of the new ring so formed may
optionally be replaced with a double bond. Alternatively, two of
the substituents on adjacent atoms of an aryl or heteroaryl ring
may optionally be replaced with a substituent of the formula
--(CRR').sub.s--X'--(C''R''').sub.d--, where s and d are
independently integers of from 0 to 3, and X' is --O--, --S--,
--S(O)--, --S(O).sub.2--, or --S(O).sub.2NR'--. The substituents R,
R', R'' and R''' are independently selected from hydrogen,
substituted or unsubstituted alkyl, substituted or unsubstituted
cycloalkyl, substituted or unsubstituted heterocycloalkyl,
substituted or unsubstituted aryl, and substituted or unsubstituted
heteroaryl.
[0035] As used herein, the term "heteroatom" or "ring heteroatom"
is meant to include oxygen (O), nitrogen (N), sulfur (S),
phosphorus (P), and silicon (Si).
[0036] A "substituent group," as used herein, means a group
selected from the following moieties: [0037] (A)-OH, --NH.sub.2,
--SH, --CN, --CF.sub.3, --NO.sub.2, oxo, halogen, unsubstituted
alkyl, unsubstituted heteroalkyl, unsubstituted cycloalkyl,
unsubstituted heterocycloalkyl, unsubstituted aryl, unsubstituted
heteroaryl, and [0038] (B) alkyl, heteroalkyl, cycloalkyl,
heterocycloalkyl, aryl, and heteroaryl, substituted with at least
one substituent selected from: [0039] (i) oxo, --OH, --NH.sub.2,
--SH, --CN, --CF.sub.3, --NO.sub.2, halogen, unsubstituted alkyl,
unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstituted
heterocycloalkyl, unsubstituted aryl, unsubstituted heteroaryl, and
[0040] (ii) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,
and heteroaryl, substituted with at least one substituent selected
from: [0041] (a) oxo, --OH, --NH.sub.2, --SH, --CN, --CF.sub.3,
--NO.sub.2, halogen, unsubstituted alkyl, unsubstituted
heteroalkyl, unsubstituted cycloalkyl, unsubstituted
heterocycloalkyl, unsubstituted aryl, unsubstituted heteroaryl, and
[0042] (b) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,
or heteroaryl, substituted with at least one substituent selected
from oxo, --OH, --NH.sub.2, --SH, --CN, --CF.sub.3, --NO.sub.2,
halogen, unsubstituted alkyl, unsubstituted heteroalkyl,
unsubstituted cycloalkyl, unsubstituted heterocycloalkyl,
unsubstituted aryl, and unsubstituted heteroaryl.
[0043] A "size-limited substituent" or "size-limited substituent
group," as used herein means a group selected from all of the
substituents described above for a "substituent group," wherein
each substituted or unsubstituted alkyl is a substituted or
unsubstituted C.sub.1-C.sub.20 alkyl, each substituted or
unsubstituted heteroalkyl is a substituted or unsubstituted 2 to 20
membered heteroalkyl, each substituted or unsubstituted cycloalkyl
is a substituted or unsubstituted C.sub.4-C.sub.8 cycloalkyl, and
each substituted or unsubstituted heterocycloalkyl is a substituted
or unsubstituted 4 to 8 membered heterocycloalkyl.
[0044] A "lower substituent" or "lower substituent group," as used
herein means a group selected from the substituents described above
for a "substituent group," wherein each substituted or
unsubstituted alkyl is a substituted or unsubstituted
C.sub.1-C.sub.8 alkyl, each substituted or unsubstituted
heteroalkyl is a substituted or unsubstituted 2 to 8 membered
heteroalkyl, each substituted or unsubstituted cycloalkyl is a
substituted or unsubstituted C.sub.5-C.sub.7 cycloalkyl, and each
substituted or unsubstituted heterocycloalkyl is a substituted or
unsubstituted 5 to 7 membered heterocycloalkyl.
[0045] The compounds of the present invention may exist as salts.
Accordingly, the present invention includes such salts. Examples of
applicable salt forms include hydrochlorides, hydrobromides,
sulfates, methanesulfonates, nitrates, maleates, acetates,
citrates, fumarates, tartrates (e.g., (+)-tartrates, (-)-tartrates
or mixtures thereof including racemic mixtures, succinates,
benzoates and salts with amino acids such as glutamic acid. These
salts may be prepared by methods known to those skilled in art.
Also included are base addition salts such as sodium, potassium,
calcium, ammonium, organic amino, or magnesium salt, or a similar
salt. When compounds of the present invention contain relatively
basic functionalities, acid addition salts can be obtained by
contacting the neutral form of such compounds with a sufficient
amount of the desired acid, either neat or in a suitable inert
solvent. Examples of acceptable acid addition salts include those
derived from inorganic acids like hydrochloric, hydrobromic,
nitric, carbonic, monohydrogencarbonic, phosphoric,
monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,
monohydrogensulfuric, hydriodic, or phosphorous acids and the like,
as well as the salts derived organic acids like acetic, propionic,
isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric,
lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic,
citric, tartaric, methanesulfonic, and the like. Also included are
salts of amino acids such as arginate and the like, and salts of
organic acids like glucuronic or galactunoric acids and the like.
Certain specific compounds of the present invention contain both
basic and acidic functionalities that allow the compounds to be
converted into either base or acid addition salts.
[0046] The neutral forms of the compounds are preferably
regenerated by contacting the salt with a base or acid and
isolating the parent compound in the conventional manner. The
parent form of the compound differs from the various salt forms in
certain physical properties, such as solubility in polar
solvents.
[0047] Certain compounds of the present invention can exist in
unsolvated forms as well as solvated forms, including hydrated
forms. In general, the solvated forms are equivalent to unsolvated
forms and are in any event encompassed within the scope of the
present invention. Certain compounds of the present invention may
exist in multiple crystalline or amorphous forms. In general, all
physical forms are equivalent or otherwise suitable for the uses
contemplated by the present invention and are within the scope of
the present invention.
[0048] Certain compounds of the present invention possess
asymmetric carbon atoms (optical centers) or double bonds; the
enantiomers, racemates, diastereomers, tautomers, geometric
isomers, stereoisometric forms that may be defined, in terms of
absolute stereochemistry, as (R)- or (S)- or, as (D)- or (L)- for
amino acids, and individual isomers are encompassed within the
scope of the present invention. The compounds of the present
invention do not include those which are known in art to be too
unstable to synthesize and/or isolate. The present invention
includes compounds in their racemic and optically pure forms.
Optically active (R)- and (S)-, or (D)- and (L)-isomers may be
prepared using chiral synthons or chiral reagents, or resolved from
mixtures using conventional techniques. When the compounds
described herein contain olefinic bonds or other centers of
geometric asymmetry, and unless specified otherwise, it is intended
that the compounds include both E and Z geometric isomers.
[0049] The compounds of the present invention may also contain
unnatural proportions of atomic isotopes at one or more of atoms
that constitute such compounds. For example, the compounds may be
radiolabeled with radioactive isotopes, such as for example tritium
(.sup.3H), iodine-125 (.sup.125I) or carbon-14 (.sup.14C). All
isotopic variations of the compounds of the present invention,
whether radioactive or not, are encompassed within the scope of the
present invention.
[0050] The term "pharmaceutically acceptable salts" is meant to
include salts of active compounds which are prepared with
relatively nontoxic acids or bases, depending on the particular
substituent moieties found on the compounds described herein. When
compounds of the present invention contain relatively acidic
functionalities, base addition salts can be obtained by contacting
the neutral form of such compounds with a sufficient amount of the
desired pharmaceutically acceptable base, either neat or in a
suitable inert solvent. Examples of pharmaceutically acceptable
base addition salts include sodium, potassium, calcium, ammonium,
organic amino, or magnesium salt, or a similar salt. When compounds
of the present invention contain relatively basic functionalities,
acid addition salts can be obtained by contacting the neutral form
of such compounds with a sufficient amount of the desired
pharmaceutically acceptable acid, either neat or in a suitable
inert solvent. Examples of pharmaceutically acceptable acid
addition salts include those derived from inorganic acids like
hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic,
phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,
monohydrogensulfuric, hydriodic, or phosphorous acids and the like,
as well as the salts derived from relatively nontoxic organic acids
like acetic, propionic, isobutyric, maleic, malonic, benzoic,
succinic, suberic, fumaric, lactic, mandelic, phthalic,
benzenesulfonic, p-tolylsulfonic, citric, tartaric,
methanesulfonic, and the like. Also included are salts of amino
acids such as arginate and the like, and salts of organic acids
like glucuronic or galactunoric acids and the like (see, for
example, Berge et al., "Pharmaceutical Salts", Journal of
Pharmaceutical Science, 1977, 66, 1-19). Certain specific compounds
of the present invention contain both basic and acidic
functionalities that allow the compounds to be converted into
either base or acid addition salts.
[0051] In addition to salt forms, the present invention provides
compounds, which are in a prodrug form. Prodrugs of the compounds
described herein are those compounds that readily undergo chemical
changes under physiological conditions to provide the compounds of
the present invention or one or more of their active metabolites.
Additionally, prodrugs can be converted to the compounds of the
present invention by chemical or biochemical methods in an ex vivo
environment. For example, prodrugs can be slowly converted to the
compounds of the present invention when placed in a transdermal
patch reservoir with a suitable enzyme or chemical reagent.
I. Anti-Inflammatory Compounds
[0052] In one aspect, the present invention provides a compound
represented by the following structural formula:
##STR00002##
[0053] In Formula (I) above, R.sup.1 is selected from substituted
or unsubstituted alkyl, substituted or unsubstituted heteroalkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocycloalkyl, substituted or unsubstituted aryl,
or substituted or unsubstituted heteroaryl.
[0054] R.sup.2, R.sup.3, and R.sup.4 are, independently, hydrogen,
substituted or unsubstituted alkyl, substituted or unsubstituted
heteroalkyl, substituted or unsubstituted cycloalkyl, substituted
or unsubstituted heterocycloalkyl, substituted or unsubstituted
aryl, or substituted or unsubstituted heteroaryl. R.sup.3 and
R.sup.4 are optionally joined with the nitrogen to which they are
attached to form a substituted or unsubstituted heterocycloalkyl or
substituted or unsubstituted heteroaryl.
[0055] L.sup.1 is a bond, substituted or unsubstituted alkylene,
substituted or unsubstituted cycloalkylene, or substituted or
unsubstituted heteroalkylene. In some embodiments, where L.sup.1 is
a bond, R.sup.2 is t-butyl, and R.sup.3 and R.sup.4 are hydrogen,
then R.sup.1 is not para-methylphenyl. In some embodiments, where
L.sup.1 is a bond, R.sup.2 is unsubstituted alkyl, and R.sup.2 and
R.sup.3 are hydrogen, then R.sup.1 is not methylphenyl.
[0056] In some embodiments, the compounds of Formula I do not
include those compounds set forth in U.S. Pat. No. 5,593,997, U.S.
Pat. No. 6,383,790, U.S. Pat. No. 5,981,533, U.S. Pat. No.
6,521,417, U.S. Pat. No. 6,921,763, U.S. Pat. No. 6,713,474, U.S.
Pat. No. 6,660,744, USP Application 2002/0156081, USP Application
2003/0073218, USP Application 2005/0085472, each of which are
incorporated by reference in their entirety for all purposes. In
some embodiments, the compounds of the invention are compounds
other than the specific compounds set forth in the aforementioned
patents and published patent applications.
[0057] In some embodiments, the compounds of the invention are the
subset of compounds of the compounds of Formula (I) that do not
displace staurosporine from a protein kinase where the compound is
present at a concentration of less than or equal to 10 .mu.M. In
some embodiments, the compounds of the invention are the subset of
compounds of the compounds of Formula (I) that do not inhibit or
significantly decrease protein kinase activity when contacted with
the kinase at a concentration of less than or equal to 10 .mu.M. In
some related embodiments, the compounds of the invention are the
subset of compounds of the compounds of Formula (I) that do not
decrease protein kinase activity more than 1%, 5%, 10%, 15%, 20%,
25%, 30%, 35%, 40%, 45%, or 50%, relative to the kinase activity in
the absence of the compound. In some related embodiments, the
compound that does not displace staurosporine as described above
and/or does not inhibit or substantially decrease the activity of a
protein kinase as described above, is a compound of Formula (I)
wherein R.sup.1 is a substituted phenyl (e.g. substituted with a
halogen), R.sup.3 and R.sup.4 are hydrogen or unsubstituted
C.sub.1-C.sub.10 alkyl, L.sup.1 is a bond or unsubstituted
C.sub.1-C.sub.10 alkylene, and R.sup.2 is unsubstituted
C.sub.1-C.sub.10 alkyl. In a further related embodiment, R.sup.1 is
a halophenyl, R.sup.3 and R.sup.4 are hydrogen or methyl, L.sup.1
is unsubstituted C.sub.1-C.sub.10 alkylene, and R.sup.2 is
unsubstituted isopropyl. In a still further related embodiment,
R.sup.3 is methyl, R.sub.3 is hydrogen, and L.sup.1 is methylene.
In some embodiments, the compound is compound 76 of Table 1,
below.
[0058] In some embodiments, R.sup.1 is substituted or unsubstituted
cycloalkyl, substituted or unsubstituted heterocycloalkyl,
substituted or unsubstituted aryl, or substituted or unsubstituted
heteroaryl.
[0059] R.sup.1 may also be an unsubstituted C.sub.3-C.sub.8
cycloalkyl, unsubstituted 4 to 8 membered heterocycloalkyl,
unsubstituted aryl, unsubstituted heteroaryl. Additionally, R.sup.1
may be C.sub.3-C.sub.s cycloalkyl, 4 to 8 membered
heterocycloalkyl, aryl, or heteroaryl, that is substituted with a
substituent selected from a halogen (e.g. fluorine and/or
chlorine), hydroxyl, sulfhydryl, cyano, nitro, unsubstituted
C.sub.1-C.sub.10 alkyl, unsubstituted 2 to 10 membered heteroalkyl,
unsubstituted C.sub.3-C.sub.8 cycloalkyl, unsubstituted 4 to 8
membered heterocycloalkyl, unsubstituted aryl, and unsubstituted
heteroaryl. In some embodiments, R.sup.1 is substituted or
unsubstituted aryl (e.g. phenyl), or substituted or unsubstituted
heteroaryl (e.g. benzothiophenyl).
[0060] In some embodiments, R.sup.1 is a substituted phenyl having
the formula:
##STR00003##
[0061] In addition to a substituted phenyl of Formula (II), R.sup.1
may be a substituted or unsubstituted benzodioxolanyl, substituted
or unsubstituted acenaphthenyl, substituted or unsubstituted
naphthyl, substituted or unsubstituted benzothiophenyl, substituted
or unsubstituted cyclopentyl, or substituted or unsubstituted
thienyl. In a related embodiment, the substituted or unsubstituted
naphthyl is a substituted or unsubstituted naphthalen-2-yl.
Alternatively, the substituted or unsubstituted naphthyl may be a
naphthalen-1-yl substituted with a substituent selected from a
hydroxyl, sulfhydryl, cyano, nitro, substituted alkyl, substituted
or unsubstituted heteroalkyl, substituted or unsubstituted
cycloalkyl, substituted or unsubstituted heterocycloalkyl,
substituted or unsubstituted aryl, or substituted or unsubstituted
heteroaryl, substituted thienyl, or unsubstituted cyclopentyl.
[0062] In some embodiments, the naphthalen-1-yl substituent is a
hydroxyl, sulfhydryl, cyano, nitro, substituted C.sub.1-C.sub.20
alkyl, substituted or unsubstituted 2 to 20 membered heteroalkyl,
substituted or unsubstituted C.sub.3-C.sub.8 cycloalkyl,
substituted or unsubstituted 4 to 8 membered heterocycloalkyl,
substituted or unsubstituted aryl, or substituted or unsubstituted
heteroaryl; thienyl substituted with a substituted or unsubstituted
alkyl; or unsubstituted cyclopentyl.
[0063] In other embodiments, R.sup.1 is unsubstituted
benzodioxolanyl or benzodioxolanyl substituted with a substituent
selected from a halogen, hydroxyl, sulfhydryl, cyano, nitro,
unsubstituted C.sub.1-C.sub.10 alkyl, unsubstituted 2 to 10
membered heteroalkyl, unsubstituted C.sub.3-C.sub.8 cycloalkyl,
unsubstituted 4 to 8 membered heterocycloalkyl, unsubstituted aryl,
and unsubstituted heteroaryl.
[0064] Alternatively, R.sup.1 is unsubstituted acenaphthenyl or
acenaphthenyl substituted with a substituent selected from a
halogen, hydroxyl, sulfhydryl, cyano, nitro, unsubstituted
C.sub.1-C.sub.10 alkyl, unsubstituted 2 to 10 membered heteroalkyl,
unsubstituted C.sub.3-C.sub.8 cycloalkyl, unsubstituted 4 to 8
membered heterocycloalkyl, unsubstituted aryl, and unsubstituted
heteroaryl.
[0065] R.sup.1 may also be a unsubstituted naphthalen-2-yl or
naphthalen-2-yl substituted with a substituent selected from a
halogen, hydroxyl, sulfhydryl, cyano, nitro, unsubstituted alkyl,
unsubstituted 2 to 10 membered heteroalkyl, unsubstituted
C.sub.3-C.sub.8 cycloalkyl, unsubstituted 4 to 8 membered
heterocycloalkyl, unsubstituted aryl, and unsubstituted
heteroaryl.
[0066] In some embodiments, R.sup.1 is naphthalen-1-yl substituted
with a substituent selected from a hydroxyl, sulfhydryl, cyano,
nitro, unsubstituted 2 to 10 membered heteroalkyl, unsubstituted
C.sub.3-C.sub.8 cycloalkyl, unsubstituted 4 to 8 membered
heterocycloalkyl, unsubstituted aryl, and unsubstituted heteroaryl.
The naphthalen-1-yl substituent may also be selected from
C.sub.1-C.sub.10 alkyl, 2 to 10 membered heteroalkyl,
C.sub.3-C.sub.8 cycloalkyl, 4 to 8 membered heterocycloalkyl, aryl,
or heteroaryl, that is substituted with a substituent selected from
a halogen, hydroxyl, sulfhydryl, cyano, nitro, unsubstituted
C.sub.1-C.sub.10 alkyl, unsubstituted 2 to 10 membered heteroalkyl,
unsubstituted C.sub.3-C.sub.8 cycloalkyl, unsubstituted 4 to 8
membered heterocycloalkyl, unsubstituted aryl, and unsubstituted
heteroaryl.
[0067] In other embodiments, R.sup.1 is thienyl substituted with an
unsubstituted C.sub.1-C.sub.10 alkyl or a C.sub.1-C.sub.10 alkyl
substituted with a substituent selected from a halogen, hydroxyl,
sulfhydryl, cyano, nitro, unsubstituted C.sub.1-C.sub.10 alkyl,
unsubstituted 2 to 10 membered heteroalkyl, unsubstituted
C.sub.3-C.sub.8 cycloalkyl, unsubstituted 4 to 8 membered
heterocycloalkyl, unsubstituted aryl, and unsubstituted
heteroaryl.
[0068] R.sup.1 may also be an unsubstituted cyclopentyl.
Alternatively, R.sup.1 is unsubstituted benzodioxolanyl or
benzodioxolanyl substituted with a substituent selected from a
halogen and unsubstituted C.sub.1-C.sub.10 alkyl; unsubstituted
acenaphthenyl or acenaphthenyl substituted with a substituent
selected from a halogen and unsubstituted C.sub.1-C.sub.10 alkyl;
unsubstituted naphthalen-2-yl or naphthalen-2-yl substituted with a
substituent selected from a halogen and unsubstituted
C.sub.1-C.sub.10 alkyl; or thienyl substituted with an
unsubstituted C.sub.1-C.sub.10 alkyl or a C.sub.1-C.sub.10 alkyl
substituted with a substituent selected from a halogen and
unsubstituted C.sub.1-C.sub.20 alkyl; or unsubstituted cyclopentyl.
R.sup.1 may additionally be a naphthalen-1-yl substituted with an
unsubstituted 2 to 10 membered heteroalkyl; or C.sub.1-C.sub.10
alkyl, or 2 to 10 membered heteroalkyl substituted with a
substituent selected from a halogen and unsubstituted
C.sub.1-C.sub.20 alkyl.
[0069] In some embodiments, R.sup.1 is unsubstituted
benzodioxolanyl; unsubstituted acenaphthenyl; unsubstituted
naphthalen-2-yl; naphthalen-2-yl substituted with a substituent
selected from a halogen and unsubstituted C.sub.1-C.sub.5 alkyl; or
thienyl substituted with an unsubstituted C.sub.1-C.sub.5 alkyl; or
unsubstituted cyclopentyl. R.sup.1 may additionally be
naphthalen-1-yl substituted with an unsubstituted 2 to 5 membered
heteroalkyl; or C.sub.1-C.sub.5 alkyl or 2 to 5 membered
heteroalkyl, that is substituted with a halogen and unsubstituted
C.sub.1-C.sub.20 alkyl.
[0070] In other embodiments, R.sup.1 is unsubstituted
benzodioxolanyl; unsubstituted acenaphthenyl; unsubstituted
naphthalen-2-yl or naphthalen-2-yl substituted with a halogen;
thienyl substituted with an unsubstituted C.sub.1-C.sub.5 alkyl; or
unsubstituted cyclopentyl.
[0071] In some embodiments, R.sup.1 is R.sup.14-substituted
C.sub.1-C.sub.20 alkyl, 2 to 20 membered unsubstituted heteroalkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocycloalkyl, R.sup.15-substituted heteroaryl,
substituted or unsubstituted fused-ring heteroaryl, or
R.sup.16-substituted aryl. R.sup.14 is oxo, --OH, halogen,
--CF.sub.3, --NH.sub.2, 2 to 20 membered substituted or
unsubstituted heteroalkyl (e.g. alkoxy), 3 to 7 membered
substituted or unsubstituted heterocycloalkyl, substituted or
unsubstituted heteroaryl, or R.sup.16-substituted aryl. R.sup.15 is
--OH, halogen, --CF.sub.3, --NH.sub.2, substituted or unsubstituted
alkyl, substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, or substituted
or unsubstituted heteroaryl. R.sup.16 is substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted heteroaryl, or
substituted or unsubstituted aryl, or substituted or unsubstituted
ortho-benzyloxy (i.e. o-benzyloxy). In some embodiments, if R.sup.1
is not unsubstituted alkyl, then R.sup.1 is optionally 2 to 20
membered substituted heteroalkyl.
[0072] In some embodiments, R.sup.1 is substituted naphthyl. In
some embodiments, R.sup.1 is substituted or unsubstituted
acenaphthenyl. In some embodiments, R.sup.1 is substituted or
unsubstituted thiophenyl-phenyl. In some embodiments, R.sup.1 is
substituted or unsubstituted naphthyl, or substituted thiophenyl,
and R.sup.2 is hydrogen or methyl.
[0073] In some embodiments, R.sup.2 is a substituted or
unsubstituted alkyl, substituted or unsubstituted heteroalkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocycloalkyl, substituted or unsubstituted aryl,
or substituted or unsubstituted heteroaryl.
[0074] R.sup.2 may be selected from a substituted or unsubstituted
C.sub.1-C.sub.20 alkyl, substituted or unsubstituted 2 to 20
membered heteroalkyl, substituted or unsubstituted C.sub.3-C.sub.8
cycloalkyl, substituted or unsubstituted 4 to 8 membered
heterocycloalkyl, substituted or unsubstituted aryl, and
substituted or unsubstituted heteroaryl. In some embodiment,
R.sup.2 is not connected to the remainder of the molecule via
nitrogen (i.e. a nitrogen-carbon bond). In some embodiments,
R.sup.2 is connected to the remainder of the molecule via a
carbon-carbon bond.
[0075] Alternatively, R.sup.2 is unsubstituted C.sub.1-C.sub.20
alkyl; unsubstituted 2 to 20 membered heteroalkyl; unsubstituted
C.sub.3-C.sub.8 cycloalkyl; unsubstituted 4 to 8 membered
heterocycloalkyl; unsubstituted aryl; unsubstituted heteroaryl; or
C.sub.1-C.sub.20 alkyl, 2 to 20 membered heteroalkyl,
C.sub.3-C.sub.8 cycloalkyl, 4 to 8 membered heterocycloalkyl, aryl,
or heteroaryl, that is substituted with a substituent selected from
a halogen, hydroxyl, sulfhydryl, cyano, oxy, nitro, unsubstituted
C.sub.1-C.sub.10 alkyl, unsubstituted 2 to 10 membered heteroalkyl,
unsubstituted C.sub.3-C.sub.8 cycloalkyl, unsubstituted 4 to 8
membered heterocycloalkyl, unsubstituted aryl, and unsubstituted
heteroaryl.
[0076] In some embodiments, R.sup.2 is an unsubstituted
C.sub.1-C.sub.10 alkyl; unsubstituted 2 to 10 membered heteroalkyl;
unsubstituted C.sub.5 cycloalkyl; unsubstituted phenyl;
unsubstituted benzyl; or C.sub.1-C.sub.10 alkyl, 2 to 10 membered
heteroalkyl, C.sub.5 cycloalkyl, phenyl, or benzyl, substituted
with a substituent selected from a halogen, hydroxyl, oxy,
sulfhydryl, cyano, nitro, unsubstituted C.sub.1-C.sub.10 alkyl,
unsubstituted 2 to 10 membered heteroalkyl, unsubstituted
C.sub.3-C.sub.8 cycloalkyl, unsubstituted 4 to 8 membered
heterocycloalkyl, unsubstituted aryl, and unsubstituted
heteroaryl.
[0077] In other embodiments, R.sup.2 is unsubstituted
C.sub.1-C.sub.10 alkyl; unsubstituted 2 to 10 membered heteroalkyl;
unsubstituted C.sub.5 cycloalkyl; unsubstituted phenyl;
unsubstituted benzyl; or C.sub.1-C.sub.10 alkyl or 2 to 10 membered
heteroalkyl, that is substituted with a substituent selected from a
halogen, hydroxyl, and oxy.
[0078] Alternatively, R.sup.2 is a hydrogen, unsubstituted alkyl,
unsubstituted heteroalkyl, unsubstituted cyclokalkyl, unsubstituted
aryl, unsubstituted arylalkyl, or unsubstituted alkylesteryl.
R.sup.2 may also be an unsubstituted C.sub.1-C.sub.20 alkyl. In
some embodiments, R.sup.2 is an unsubstituted C.sub.1-C.sub.10
alkyl. In other embodiments, R.sup.2 is tertiary butyl.
[0079] In some embodiments, R.sup.2 is R.sup.10-substituted
C.sub.1-C.sub.20 alkyl, 2 to 20 membered substituted or
unsubstituted heteroalkyl, W.sup.1-substituted C.sub.3-C.sub.8
cycloalkyl, R.sup.11-substituted C.sub.3-C.sub.8 heterocycloalkyl,
R.sup.12-substituted heteroaryl, or Rn-substituted aryl. R.sup.10
is oxo, --OH, halogen, --CF.sub.3, --NH.sub.2, 2 to 20 membered
substituted or unsubstituted heteroalkyl (e.g. substituted or
unsubstituted alkoxy), 3 to 7 membered substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted heteroaryl, or
R.sup.13-substituted aryl. R.sup.11 is oxo, --OH, halogen,
--CF.sub.3, --NH.sub.2, substituted or unsubstituted alkyl,
substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, or substituted
or unsubstituted heteroaryl. R.sup.12 is --OH, halogen, --CF.sub.3,
--NH.sub.2, substituted or unsubstituted alkyl, substituted or
unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted heterocycloalkyl, substituted or
unsubstituted aryl, or substituted or unsubstituted heteroaryl.
R.sup.13 is --OH, --NH.sub.2, substituted or unsubstituted
cycloalkyl, substituted or unsubstituted heterocycloalkyl,
substituted or unsubstituted heteroaryl, or substituted or
unsubstituted aryl. In some related embodiments, if R.sup.3 and
R.sup.4 are hydrogen and R.sup.1 is 4-methylphenyl, then R.sup.2 is
not 2-ethylacetyl. In other related embodiments, if R.sup.3 and
R.sup.4 are hydrogen and R.sup.1 is methylphenyl or ethylphenyl,
then R.sup.2 is not alkylacetyl.
[0080] In certain embodiments, R.sup.3 and R.sup.4 are
independently selected from a hydrogen, substituted or
unsubstituted alkyl, and substituted or unsubstituted heteroalkyl.
In some embodiments, R.sup.3 and R.sup.4 are, independently, a
hydrogen, substituted or unsubstituted C.sub.1-C.sub.10 alkyl, or
substituted or unsubstituted 2 to 10 membered heteroalkyl. R.sup.3
and R.sup.4 may also be independently selected from a hydrogen,
unsubstituted C.sub.1-C.sub.10 alkyl, and unsubstituted 2 to 10
membered heteroalkyl. Alternatively, R.sup.3 and R.sup.4 are,
independently, a hydrogen or methyl. In some embodiments, R.sup.3
and R.sup.4 are hydrogen.
[0081] In some embodiments, R.sup.3 and R.sup.4 are, independently,
substituted or unsubstituted alkyl, substituted or unsubstituted
heteroalkyl, substituted or unsubstituted cycloalkyl, substituted
or unsubstituted heterocycloalkyl, substituted or unsubstituted
aryl, or substituted or unsubstituted heteroaryl. In some
embodiments, R.sup.3 and R.sup.4 are unsubstituted alkyl.
[0082] In some embodiments, R.sup.1 is phenyl substituted with
halogen, methyl, --OH, --CF.sub.3, --OCH.sub.3, or --NH.sub.2;
R.sup.2 is C.sub.1-C.sub.10 unsubstituted alkyl; R.sup.3 and
R.sup.4 are independently hydrogen or methyl; and L.sup.1 is
methylene. R.sup.1 may also be phenyl substituted with halogen
(e.g. chlorine). In some embodiments, R.sup.1 is
ortho-chlorophenyl. R.sup.1 may also be phenyl substituted with
halogen, --OH, --CF.sub.3, --OCH.sub.3, or --NH.sub.2. In some
embodiments, R.sup.3 and R.sup.4 are hydrogen. In some embodiments,
R.sup.3 is hydrogen and R.sup.4 is methyl.
[0083] L.sup.1 may be a bond, unsubstituted C.sub.1-C.sub.10
alkylene, or C.sub.1-C.sub.10 alkylene substituted with
unsubstituted alkyl, unsubstituted heteroalkyl, unsubstituted
cycloalkyl, unsubstituted heteroalkyl, unsubstituted aryl, or
unsubstituted heteroaryl. In some embodiments, L.sup.1 is
methylene.
[0084] L.sup.1 may also be a bond, substituted or unsubstituted
alkylene, or substituted or unsubstituted heteroalkylene, wherein
if R.sup.1 is a substituted phenyl, substituted or unsubstituted
benzodioxolanyl, or substituted or unsubstituted cyclopentyl, then
L.sup.1 is substituted or unsubstituted alkylene, or substituted or
unsubstituted heteroalkylene.
[0085] In some embodiments, L.sup.1 is a bond or substituted or
unsubstituted alkylene, wherein if R.sup.1 is a substituted phenyl,
substituted or unsubstituted benzodioxolanyl, or unsubstituted
cyclopentyl, then L.sup.1 is substituted or unsubstituted
alkylene.
[0086] In other embodiments, L.sup.1 is a bond or unsubstituted
C.sub.1-C.sub.20 alkylene, wherein if R.sup.1 is a phenyl,
benzodioxolanyl, or cyclopentyl, then L.sup.1 is an unsubstituted
C.sub.1-C.sub.20 alkylene. In other embodiments, L.sup.1 is a bond
or unsubstituted C.sub.1-C.sub.20 alkylene, wherein if R.sup.1 is a
phenyl, benzodioxolanyl, or cyclopentyl, then L.sup.1 is an
unsubstituted C.sub.2-C.sub.20 alkylene. In other embodiments,
L.sup.1 is a bond or unsubstituted C.sub.1-C.sub.20 alkylene,
wherein if R.sup.1 is a phenyl, benzodioxolanyl, or cyclopentyl,
then L.sup.1 is an unsubstituted C.sub.3-C.sub.20 alkylene.
[0087] Alternatively, L.sup.1 is a bond or unsubstituted
C.sub.1-C.sub.10 alkylene, wherein if R.sup.1 is a phenyl,
benzodioxolanyl, or cyclopentyl, then L.sup.1 is an unsubstituted
C.sub.1-C.sub.10 alkylene.
[0088] L.sup.1 may also be a bond or unsubstituted C.sub.1-C.sub.5
alkylene, wherein if R.sup.1 is a phenyl, benzodioxolanyl, or
cyclopentyl, then L.sup.1 is an unsubstituted C.sub.1-C.sub.5
alkylene. Alternatively, L.sup.1 may be a bond or methylene,
wherein if R.sup.1 is a phenyl, benzodioxolanyl, or cyclopentyl,
then L.sup.1 is a methylene.
[0089] In some embodiments, L.sup.1 is substituted or unsubstituted
[1,1]cycloalkylene, such as substituted or unsubstituted
[1,1]-cyclopropylene. In some embodiments, L.sup.1 is unsubstituted
[1,1]cycloalkylene, such as unsubstituted [1,1]-cyclopropylene.
[0090] In an exemplary embodiment, R.sup.5, R.sup.6, R.sup.7,
R.sup.8 and R.sup.9 of Formula (II), above, are independently
selected from a hydrogen; halogen; hydroxyl; sulfhydryl; cyano;
nitro; substituted or unsubstituted alkyl; substituted or
unsubstituted heteroalkyl; substituted or unsubstituted cycloalkyl;
substituted or unsubstituted heterocycloalkyl; substituted or
unsubstituted aryl; and substituted or unsubstituted
heteroaryl.
[0091] In some embodiments, R.sup.5, R.sup.7, and R.sup.9 are
independently selected from a hydrogen; halogen; hydroxyl;
sulfhydryl; cyano; nitro; substituted or unsubstituted alkyl;
substituted or unsubstituted heteroalkyl; substituted or
unsubstituted cycloalkyl; substituted or unsubstituted
heterocycloalkyl; substituted or unsubstituted aryl; and
substituted or unsubstituted heteroaryl. R.sup.6 and R.sup.8 are
independently selected from a hydrogen, halogen, hydroxyl,
sulfhydryl, cyano, nitro, substituted or unsubstituted alkyl,
unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted heterocycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted heteroaryl, and
heteroalkyl substituted with a substituted aryl. In certain
embodiments, R.sup.5, R.sup.6, R.sup.8, and R.sup.9 are not
simultaneously hydrogen.
[0092] In other embodiments, R.sup.5, R.sup.7, and R.sup.9 are,
independently, hydrogen, halogen, hydroxyl, sulfhydryl, cyano,
nitro, substituted or unsubstituted C.sub.1-C.sub.20 alkyl,
substituted or unsubstituted 2 to 20 membered heteroalkyl,
substituted or unsubstituted C.sub.3-C.sub.8 cycloalkyl,
substituted or unsubstituted 4 to 8 membered heterocycloalkyl,
substituted or unsubstituted aryl, or substituted or unsubstituted
heteroaryl.
[0093] Alternatively, R.sup.5, R.sup.7, and R.sup.9 are,
independently, hydrogen; halogen; hydroxyl; sulfhydryl; cyano;
nitro; unsubstituted C.sub.1-C.sub.10 alkyl; unsubstituted 2 to 10
membered heteroalkyl; unsubstituted C.sub.3-C.sub.8 cycloalkyl;
unsubstituted 4 to 8 membered heterocycloalkyl; unsubstituted aryl;
unsubstituted heteroaryl; or C.sub.1-C.sub.10 alkyl, 2 to 20
membered heteroalkyl, C.sub.3-C.sub.8 cycloalkyl, 4 to 8 membered
heterocycloalkyl, aryl, or heteroaryl, that is substituted with a
substituent selected from a halogen, hydroxyl, sulfhydryl, cyano,
nitro, unsubstituted C.sub.1-C.sub.10 alkyl, unsubstituted 2 to 10
membered heteroalkyl, unsubstituted C.sub.3-C.sub.8 cycloalkyl,
unsubstituted 4 to 8 membered heterocycloalkyl, unsubstituted
heteroaryl, unsubstituted aryl, and aryl substituted with a halogen
or unsubstituted heteroaryl.
[0094] R.sup.5, R.sup.7, and R.sup.9 may also be, independently,
hydrogen; halogen; hydroxyl; unsubstituted C.sub.1-C.sub.10 alkyl;
unsubstituted 2 to 10 membered heteroalkyl; unsubstituted
C.sub.5-C.sub.8 cycloalkyl; unsubstituted 5 to 8 membered
heterocycloalkyl; unsubstituted aryl; unsubstituted heteroaryl; or
C.sub.1-C.sub.10 alkyl or 2 to 10 membered heteroalkyl, that is
substituted with a substituent selected from an unsubstituted aryl,
unsubstituted heteroaryl, and aryl substituted with a halogen.
[0095] In another embodiment, R.sup.5, R.sup.7, and R.sup.9 are
independently selected from a hydrogen, halogen, unsubstituted
C.sub.1-C.sub.10 alkyl, and unsubstituted 2 to 10 membered
heteroalkyl. R.sup.5, R.sup.7, and R.sup.9 may also independently
be a hydrogen, halogen, methyl, or methoxy.
[0096] In another embodiment, R.sup.6 and R.sup.8 are,
independently, hydrogen, halogen, hydroxyl, sulfhydryl, cyano,
nitro, substituted or unsubstituted C.sub.1-C.sub.20 alkyl,
unsubstituted 2 to 20 membered heteroalkyl, substituted or
unsubstituted C.sub.3-C.sub.8 cycloalkyl, substituted or
unsubstituted 4 to 8 membered heterocycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted heteroaryl, or 2
to 20 membered heteroalkyl substituted with a substituted aryl.
[0097] In some embodiments, R.sup.6 and R.sup.8 are, independently,
hydrogen; halogen; hydroxyl; sulfhydryl; cyano; nitro;
unsubstituted C.sub.1-C.sub.10 alkyl; unsubstituted 2 to 10
membered heteroalkyl; unsubstituted C.sub.3-C.sub.8 cycloalkyl;
unsubstituted 4 to 8 membered heterocycloalkyl; unsubstituted aryl;
unsubstituted heteroaryl; 2 to 10 membered heteroalkyl substituted
with a substituted aryl; or C.sub.1-C.sub.10 alkyl, C.sub.3-C.sub.8
cycloalkyl, 4 to 8 membered heterocycloalkyl, aryl, or heteroaryl,
substituted with a substituent selected from a halogen, hydroxyl,
sulfhydryl, cyano, nitro, unsubstituted C.sub.1-C.sub.10 alkyl,
unsubstituted 2 to 10 membered heteroalkyl, unsubstituted
C.sub.3-C.sub.8 cycloalkyl, unsubstituted 4 to 8 membered
heterocycloalkyl, unsubstituted aryl, and unsubstituted
heteroaryl.
[0098] In other embodiments, R.sup.6 and R.sup.8 are,
independently, hydrogen; halogen; hydroxyl; unsubstituted
C.sub.1-C.sub.10 alkyl; unsubstituted 2 to 10 membered heteroalkyl;
unsubstituted C.sub.3-C.sub.8 cycloalkyl; unsubstituted 4 to 8
membered heterocycloalkyl; unsubstituted aryl; unsubstituted
heteroaryl; 2 to 10 membered heteroalkyl substituted with a
haloaryl; or C.sub.1-C.sub.10 alkyl substituted with a substituent
selected from an unsubstituted aryl, haloaryl, or unsubstituted
heteroaryl.
[0099] R.sup.6 and R.sup.8 may also be selected from a hydrogen,
halogen, unsubstituted C.sub.1-C.sub.10 alkyl, unsubstituted 2 to
10 membered heteroalkyl, or 2 to 5 membered heteroalkyl substituted
with a haloaryl.
[0100] Alternatively, R.sup.6 and R.sup.8 are independently
selected from a hydrogen, halogen, methyl, methoxy, and methoxy
substituted with a haloaryl.
[0101] In some embodiments, each substituted group described above
in the compounds of Formulae (I) and/or (II) is substituted with at
least one substituent group. Fore example, in some embodiments,
each substituted alkyl, substituted heteroalkyl, substituted
cycloalkyl, substituted heterocycloalkyl, substituted aryl,
substituted heteroaryl, substituted alkylene, substituted
cycloalkylene, and/or substituted heteroalkylene, described above
in the compounds of Formulae (I) and/or (II) are substituted with
at least one substituent group. In other embodiments, at least one
or all of these groups are substituted with at least one
size-limited substituent group. Alternatively, at least one or all
of these groups are substituted with at least one lower substituent
group.
[0102] In other embodiments of the compounds of Formulae (I) and/or
(II), each substituted or unsubstituted alkyl is a substituted or
unsubstituted C.sub.1-C.sub.20 alkyl, each substituted or
unsubstituted heteroalkyl is a substituted or unsubstituted 2 to 20
membered heteroalkyl, each substituted or unsubstituted cycloalkyl
is a substituted or unsubstituted C.sub.3-C.sub.8 cycloalkyl, each
substituted or unsubstituted heterocycloalkyl is a substituted or
unsubstituted 4 to 8 membered heterocycloalkyl, each substituted or
unsubstituted alkylene is a substituted or unsubstituted
C.sub.1-C.sub.20 alkylene, each substituted or unsubstituted
cycloalkylene is a C.sub.3-C.sub.8 cycloalkylene, and/or each
substituted or unsubstituted heteroalkylene is a substituted or
unsubstituted 2 to 20 membered heteroalkylene.
[0103] Alternatively, each substituted or unsubstituted alkyl is a
substituted or unsubstituted C.sub.1-C.sub.8 alkyl, each
substituted or unsubstituted heteroalkyl is a substituted or
unsubstituted 2 to 8 membered heteroalkyl, each substituted or
unsubstituted cycloalkyl is a substituted or unsubstituted
C.sub.5-C.sub.7 cycloalkyl, each substituted or unsubstituted
heterocycloalkyl is a substituted or unsubstituted 5 to 7 membered
heterocycloalkyl, each substituted or unsubstituted alkylene is a
substituted or unsubstituted C.sub.1-C.sub.8 alkylene, each
substituted or unsubstituted cycloalkylene is a C.sub.3-C.sub.6
cycloalkylene, and/or each substituted or unsubstituted
heteroalkylene is a substituted or unsubstituted 2 to 8 membered
heteroalkylene.
[0104] In some embodiments, the compounds of the invention are the
subset of compounds set forth in Table 1 below, with the exception
of compound 13 and/or compound 57. Although compound 57 does not
fall within the scope of Formula (I), in some embodiments compound
57 of Table 1 is useful in the methods of the present invention. In
some embodiments, the compounds of the invention are the subset of
compounds set forth in Table 1 below, with the exception of
compound 57. In some embodiments, the compounds of the invention
are the subset of compounds in Table 1 that are set forth as
"active." In some embodiments, the compounds of the invention do
not include those compounds set forth in Table 2 below. One skilled
in the art, using the methods described herein and/or methods well
known in the art, can easily determine which compounds are useful
as anti-inflammatory compounds (see e.g. section III below)
II. Exemplary Synthesis
[0105] The compounds of the invention are synthesized by an
appropriate combination of generally well known synthetic methods.
Techniques useful in synthesizing the compounds of the invention
are both readily apparent and accessible to those of skill in the
relevant art in view of the disclosure herein. The discussion below
is offered to illustrate certain of the diverse methods available
for use in assembling the compounds of the invention. However, the
discussion is not intended to define the scope of reactions or
reaction sequences that are useful in preparing the compounds of
the present invention.
##STR00004##
[0106] In Scheme 1, R.sup.1 and L.sup.1 are as defined above. In
step (i), carbonyl compound I is reacted with NaH and malononitrile
in THF, followed by (ii) treatment with NaHCO.sub.3, and dimethyl
sulfate in dioxane/H.sub.2O to form the corresponding alkene 2.
Cyclization of 2 is accomplished by refluxing in the presence of
triethylamine and tert-butylhydrazine hydrochloride in ethanol to
from the pyrazole 3. Finally, 3 is reacted with formamide to yield
the pyrazolopyrimidine 4.
##STR00005##
[0107] The amine substituent of 4 may be derivatized by reacting
with the appropriate acyl chloride (R'--C(O)--Cl) in the presence
of pyridine (v) (Scheme 2). The resulting amide may be reduced by
refluxing with LiAlH.sub.4 in dry tetrahydrofuran to form 6. In
scheme 2, R' represents substituted or unsubstituted alkyl,
substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, and
substituted or unsubstituted heteroaryl. In some embodiments, the
--CH.sub.2--R' moiety is R.sup.3, which is defined above.
[0108] For a more detailed discussion of useful synthesis
procedures, see Bishop et al., J. Am. Chem. Soc. 121, 627-631
(1999) and Bishop et al., Current Biology, 8: 257-266 (1998).
III. Methods of Identifying an Anti-Inflammatory
Pyrazolopyrimidine
[0109] In another aspect, the present invention provides a method
for identifying anti-inflammatory pyrazolopyrimidine compounds. The
method includes contacting a candidate anti-inflammatory
pyrazolopyrimidine compound with a cell culture assay. The cell
culture assay may include more than one type of cell, also referred
to herein as a cell culture assay combination. The cell culture
assay combination may include, for example and without limitation,
human endothelial cells in an inflammatory state with or without
peripheral blood mononuclear cells. In one type of cell culture
assay, changes in the expression of at least two different gene
products relevant to an inflammatory state are detected. The change
in the expression of the at least two different gene products is
compared to the expression of those gene products in the absence of
the candidate anti-inflammatory pyrazolopyrimidine compound, and if
the candidate compound alters the expression of one or more of such
gene products in a manner consistent with anti-inflammatory action
(for example, by decreasing the expression of a gene product that
induces an inflammatory state or is characteristic of an
inflammatory state or by increasing the expression of a gene
product that inhibits inflammation), then the candidate compound is
thereby identified as an anti-inflammatory pyrazolopyrimidine
compound. Cell culture assays and methods for analyzing the data
obtained by them useful in this aspect of the invention are
described in PCT publication Nos. 05/023987; 04/094992; 04/094609;
04/022711; 03/023753; and 01/067103; and U.S. Pat. Nos. 6,656,695;
and 6,763,307, each of which is incorporated herein by
reference.
[0110] In some embodiments, the gene products analyzed are the
products of a gene selected from E-selectin, ICAM-1, VCAM-1,
CXLC9/MIG, IL-6, CXCL8/IL-8, HLA-DR, CCL2/MCP-1, P-selectin,
VEGFR2, CD87/uPAR, CD55, CCL26/Eotaxin-3, CD14, CD40, CD69, CD31,
CD38, CD142/TF, IL-1.alpha., M-CSF, CD141/TM, Endothelin-1, LDLR,
CXCL10/IP-10, CD3, and IL-2. Additional gene products that may be
analyzed in such assays are discussed in detail, for example, in
the above cited PCT patent publications and issued US patents, each
of which has been incorporated by reference in their entirety for
all purposes. Other exemplary targets and pathways that may be
detected in such cell culture assays are illustrated in FIGS. 1A
and 1B below.
[0111] The inflammatory state of the cells in the cell culture
assay may be achieved by adding to the culture one or more
inflammatory factors selected from TNF-.alpha., TNF-.beta., IL-1,
IL-2, IL-4, IL-12, IL-13, Staphylococcal Enterotoxin B (SEB),
Staphylococcal Enterotoxin E (SEE), toxic shock syndrome toxin
(TSST), lipopolysaccharide (LPS), anti-CD3 antibody, anti-T cell
receptor antibody, histamine and IFN-.gamma.. The inflammatory
factors are typically added in an amount and for a time sufficient
to induce said inflammatory state. In some embodiments, at least
one, two or three inflammatory factors are added. Additional
inflammatory factors are discussed in detail, for example, in the
above cited PCT patent publications and issued US patents.
[0112] In certain embodiments, activities of a candidate
pyrazolopyrimidine compound are compared to a known
anti-inflammatory compound. Pyrazolopyrimidine compounds with novel
activity profiles or with activity profiles similar to other
anti-inflammatory compounds may thereby be detected.
[0113] In an exemplary embodiment, a Biologically Multiplexed
Activity Profiling system, known under the mark BioMAP.TM. system,
is employed. Useful BioMAP systems are discussed in detail, for
example, in the above cited PCT patent publications and issued US
patents and in Kunkel et al., ASSAY and Drug Development
Technologies, 2: 431-441 (2004), incorporated herein by
reference.
[0114] Methods for selecting cells and culture conditions for
screening candidate anti-inflammatory pyrazolopyrimidines,
identification of an optimized set of discrete parameters to be
measured, the use of BioMAP analysis for rapid identification and
characterization of candidates, and the like, are within the skill
of those having knowledge in the biological and/or biochemical arts
in view of the disclosure herein and that of the above cited PCT
patent publications and issued US patents. In some embodiments, a
large number of cellular pathways, and the rapid identification of
compounds that cause cellular responses, are simultaneously
screened.
[0115] Agents are screened for biological activity by adding an
agent to be screened to at least one and usually a plurality of
assay combinations to form a panel of assay combinations, usually
in conjunction with assay combinations lacking agent. The change in
parameter readout in response to agent is measured, desirably
normalized, and the resulting BioMAP readout may then be evaluated
by comparison to reference BioMAP readouts. The reference BioMAP
readouts may include basal readouts in the presence and absence of
the factors, BioMAP readouts obtained with other agents, which may
or may not include known inhibitors of known pathways. Agents of
interest for analysis include any biologically active molecule with
the capability of modulating, directly or indirectly, the phenotype
of interest of a cell of interest.
[0116] Various methods can be utilized for quantifying the presence
of the selected markers. For measuring the amount of a molecule
that is present, a convenient method is to label a molecule with a
detectable moiety, which may be fluorescent, luminescent,
radioactive, enzymatically active, etc., particularly a molecule
specific for binding to the parameter with high affinity
Fluorescent moieties are readily available for labeling virtually
any biomolecule, structure, or cell type. Immunofluorescent
moieties can be directed to bind not only to specific proteins but
also specific conformations, cleavage products, or site
modifications like phosphorylation. Individual peptides and
proteins can be engineered to autofluoresce, e.g. by expressing
them as green fluorescent protein chimeras inside cells (for a
review see Jones et al. (1999) Trends Biotechnol., 17(12):477-81).
Thus, antibodies can be genetically modified to provide a
fluorescent dye as part of their structure. Applicable methods are
generally known in art, and are discussed in detail, for example,
in the above cited PCT patent publications and issued US
patents.
[0117] In certain embodiments, human umbilical vein endothelial
cells (HUVEC) and/or peripheral blood mononuclear cells (PBMC) are
contacted with different combinations of cytokines to induce an
inflammatory response. Exemplary cytokine combinations are
illustrated in FIG. 1B. The inflammatory response is assessed by
detecting the presence of the gene products shown in FIG. 1B.
IV. Methods for Treating or Preventing a Disorder Characterized by
Inflammation
[0118] In another aspect, the present invention provides a method
for treating or preventing a disorder characterized by
inflammation, said method including administering to a subject a
therapeutically effective amount of a compound of the present
invention.
[0119] In certain embodiments, the inflammatory process that occurs
in response to or is an underlying cause of infection, trauma,
autoimmune disease, cardiovascular disease, neoplasia, hyperplasia,
addiction, infection, obesity, cellular degeneration, apoptosis, or
senescence, or differentiation.
[0120] The disorder may be selected from the group consisting of
vasculitis, multiple sclerosis, diabetes, inflammatory bowel
disease (e.g. Crohn's disease and ulcerative colitis), psoriasis,
arthritis (e.g. rheumatoid arthritis, spondyloarthopathies, gouty
arthritis, osteoarthritis, and juvenile arthritis), asthma, stroke,
atherosclerosis (including atherosclerotic plaque rupture),
restenosis, and lupus (including systemic lupus erythematosus).
[0121] Other disorders associated with abnormal inflammation
include, for example, headaches (e.g. migraine headaches),
bronchitis, menstrual cramps, tendonitis, gastritis, vascular
diseases, uveitis, Sjogren's disease, sclerodoma, nephrotic
syndrome, swelling occurring after injury, myocardial ischemia,
fever (including rheumatic fever and fever associated with
influenza and other viral infections), common cold, dysmenorrhea,
emphysema, acute respiratory distress syndrome, chronic obstructive
pulmonary disease, Alzheimer's disease, organ transplant toxicity,
cachexia, allergic reactions, allergic contact hypersensitivity,
cancer (such as solid tumor cancer including colon cancer, breast
cancer, lung cancer and prostrate cancer; hematopoietic
malignancies including leukemias and lymphomas; Hodgkin's disease;
aplastic anemia, skin cancer and familiar adenomatous polyposis),
tissue ulceration, peptic ulcers, regional enteritis, ulcerative
colitis, diverticulitis, recurrent gastrointestinal lesion,
gastrointestinal bleeding, coagulation, synovitis, gout, ankylosing
spondylitis, restenosis, periodontal disease, epidermolysis
bullosa, osteoporosis, loosening of artificial joint implants,
aortic aneurysm (including abdominal aortic aneurysm and brain
aortic aneurysm), periarteritis nodosa, congestive heart failure,
myocardial infarction, cerebral ischemia, head trauma, spinal cord
injury, neuralgia, neuro-degenerative disorders (acute and
chronic), autoimmune disorders, Huntington's disease, Parkinson's
disease, migraine, depression, peripheral neuropathy, pain
(including low back and neck pain, headache and toothache),
gingivitis, cerebral amyloid angiopathy, nootropic or cognition
enhancement, amyotrophic lateral sclerosis, multiple sclerosis,
ocular angiogenesis, corneal injury, macular degeneration,
conjunctivitis, abnormal wound healing, muscle or joint sprains or
strains, tendonitis, skin disorders (such as psoriasis, eczema,
scleroderma and dermatitis), myasthenia gravis, polymyositis,
myositis, bursitis, burns, diabetes (including types I and II
diabetes, diabetic retinopathy, neuropathy and nephropathy, insulin
dependent diabetes mellitus (IDDM)), tumor invasion, tumor growth,
tumor metastasis, corneal scarring, scleritis, immunodeficiency
diseases (such as AIDS in humans and FLV, FIV in cats), sepsis,
premature labor, hypoprothrombinemia, hemophilia, thyroiditis,
sarcoidosis, Behcet's syndrome, hypersensitivity, kidney disease,
Rickettsial infections (such as Lyme disease, Erlichiosis),
Protozoan diseases (such as malaria, giardia, coccidia),
reproductive disorders (preferably in livestock) and septic
shock.
[0122] Treatment may be conducted in a mammal (e.g. a human, cat,
dog, or livestock), and the treatment method comprises
administering a therapeutically effective amount of a compound of
Formula (I) or a pharmaceutically acceptable salt thereof in
combination with a pharmaceutically acceptable carrier.
V. Anti-Inflammatory Activity
[0123] As discussed above, the present invention provides
anti-inflammatory compounds useful in the treatment of diseases and
conditions in which inflammation is involved in disease progression
or the manifestation of symptoms of the disease or condition.
[0124] In one embodiment, the compounds of the invention are the
subset of compounds of Formula (I) that are T cell activation
inhibitors. T-cell activation inhibitors are those compounds that
strongly inhibit T cell activation in a cell-based assay (e.g. a
BioMAP assay as described above, where, for example, a superantigen
is used to stimulate T cells). The phrase "strongly inhibit T cell
activation," as used herein, refers to compounds that inhibit the
expression of at least one gene product selected from E-selectin,
ICAM-1, VCAM-1, CXLC9/MIG, CXCL8/IL-8, CCL2/MCP-1, CD87/uPAR,
CCL26/Eotaxin-3, CD40, CD69, CD38, CD142/TF, CXCL10/IP-10, or IL-2
by at least 50% in a cell-based assay. T-cell activation inhibitors
may be especially useful in treating the inflammation underlying
arthritis, psoriasis, vasculitis, multiple sclerosis, IBD, asthma,
atherosclerosis, or other inflammatory disease. Illustrative
pyrazolopyrimidine T cell activation inhibitor compounds of this
embodiment include compounds 10, 13, 15, 17, 41, 48, and 54 in
Table 1 below.
[0125] In another embodiment, the compounds of the invention are
the subset of compounds of Formula (I0 that are monocyte activation
and recruitment inhibitors. Monocyte activation inhibitors weakly
inhibit T cell activation, strongly inhibit monocyte activation,
and strongly inhibit MCP-1 and IL-8 production from multiple cell
types including but not limited to endothelial cells, fibroblasts,
and smooth muscle cells. The phrase "weakly inhibit T cell
activation," as used herein, refers to compounds that inhibit the
expression of at least one gene product selected from E-selectin,
ICAM-1, VCAM-1, CXLC9/MIG, CXCL8/IL-8, CCL2/MCP-1, CD87/uPAR,
CCL26/Eotaxin-3, CD40, CD69, CD38, CD142/TF, CXCL10/IP-10, or IL-2
by less than 50% in a cell-based assay. The phrase "strongly
inhibit monocyte activation," as used herein, refers to compounds
that inhibit the expression of at least one gene product selected
from E-selectin, ICAM-1, VCAM-1, CXLC9/MIG, IL-6, CXCL8/IL-8,
CCL2/MCP-1, CD14, CD40, CD69, CD142/TF, IL-1.alpha., M-CSF,
CD141/TM, or CXCL10/IP-10 by at least 50% in a cell-based assay.
The phrase "strongly inhibit strongly inhibit MCP-1," as used
herein, refers to compounds that inhibit the expression of MCP-1
gene product by at least 50% in a cell-based assay. The phrase
"strongly inhibit strongly inhibit IL-8," as used herein, refers to
compounds that inhibit the expression of IL-8 gene product by at
least 50% in a cell-based assay. Monocyte activation inhibitors may
be especially useful in treating the inflammation underlying
multiple diseases including rheumatoid arthritis, ankylosing
spondylitis, psoriatic arthritis, multiple sclerosis, psoriasis,
inflammatory bowel disease, atherosclerosis, diabetes, lupus
nephritis, and vasculitis. Illustrative pyrazolopyrimidine monocyte
activation inhibitor compounds of this embodiment include compounds
30, 31, 36, 59, 62, 63, 64, 65, 72, and 76 in Table 1 below.
[0126] In another embodiment, the compounds of the invention are
the subset of compounds of Formula (I) that are gamma interferon
signaling inhibitors. Gamma interferon signaling inhibitors weakly
inhibit T cell activation (see above) and monocyte activation (see
above) but inhibit IFN-gamma signaling and collagen production. The
phrase "weakly inhibit monocyte activation," as used herein, refers
to compounds that inhibit the expression of at least one gene
product selected from E-selectin, ICAM-1, VCAM-1, CXLC9/MIG, IL-6,
CXCL8/IL-8, CCL2/MCP-1, CD14, CD40, CD69, CD142/TF, IL-1.alpha.,
M-CSF, CD141/TM, or CXCL10/IP-10 by less than 50% in a cell-based
assay. Gamma interferon signaling inhibitors may be especially
useful in treating the inflammation underlying multiple diseases
including rheumatoid arthritis, systemic sclerosis, COPD, asthma,
atherosclerosis, cirrhosis, pulmonary fibrosis, sarcoidosis,
keloids, and renal fibrosis. Illustrative pyrazolopyrimidine
interferon gamma signaling inhibitor compounds of this embodiment
include compounds 7, 8, 9, 11, 12, and 19 in Table 1 below.
VI. Pharmaceutical Compositions
[0127] In another aspect, the present invention provides
pharmaceutical compositions comprising a pharmaceutically
acceptable excipient and a compound of the present invention, such
as a compound within the scope of Formula (I) provided above.
[0128] The compounds of the present invention can be prepared and
administered in a wide variety of oral, parenteral and topical
dosage forms. Oral preparations include tablets, pills, powder,
dragees, capsules, liquids, lozenges, gels, syrups, slurries,
suspensions, etc., suitable for ingestion by the patient. The
compounds of the present invention can also be administered by
injection, that is, intravenously, intramuscularly,
intracutaneously, subcutaneously, intraduodenally, or
intraperitoneally. Also, the compounds described herein can be
administered by inhalation, for example, intranasally.
Additionally, the compounds of the present invention can be
administered transdermally. The anti-inflammatory
pyrazolopyrimidines of this invention can also be administered by
intraocular, intravaginal, and intrarectal routes including via
suppositories, insufflation, powders and aerosol formulations (for
examples of steroid inhalants, see Rohatagi, J Clin. Pharmacol.
35:1187-1193, 1995; Tjwa, Ann. Allergy Asthma Immunol. 75:107-111,
1995). Accordingly, the present invention also provides
pharmaceutical compositions comprising a pharmaceutically
acceptable carrier or excipient and either a compound of Formula
(I), or a pharmaceutically acceptable salt of a compound of Formula
(I).
[0129] The compounds of the present invention may also be
administered using an anti-inflammatory pyrazolopyrimidine coated
stent. The stent may be inserted into a vessel, acting as a
scaffold to provide structural support for the vessel thereby
holding the vessel open and improving blood flow through the
vessel. An anti-inflammatory pyrazolopyrimidine coated stent refers
to a stent that has been coated with anti-inflammatory
pyrazolopyrimidine and optionally additional agents, such as an
appropriate pharmaceutical excipient. The anti-inflammatory
pyrazolopyrimidine coated stent typically allows the
anti-inflammatory pyrazolopyrimidine to be released over time into
surrounding tissue. One skilled in the art understands that the
stent may be adsorbed to, impregnated with, covalently attached to,
or ionically bonded to the anti-inflammatory
pyrazolopyrimidine.
[0130] For preparing pharmaceutical compositions from the compounds
of the present invention, pharmaceutically acceptable carriers can
be either solid or liquid. Solid form preparations include powders,
tablets, pills, capsules, cachets, suppositories, and dispersible
granules. A solid carrier can be one or more substances, which may
also act as diluents, flavoring agents, binders, preservatives,
tablet disintegrating agents, or an encapsulating material. Details
on techniques for formulation and administration are well described
in the scientific and patent literature, see, e.g., the latest
edition of Remington's Pharmaceutical Sciences, Maack Publishing
Co, Easton Pa. ("Remington's").
[0131] In powders, the carrier is a finely divided solid, which is
in a mixture with the finely divided active component. In tablets,
active component is mixed with a carrier having the necessary
binding properties in suitable proportions and compacted in the
shape and size desired.
[0132] The powders and tablets preferably contain from 5% or 10% to
70% of active compound. Suitable carriers are magnesium carbonate,
magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch,
gelatin, tragacanth, methylcellulose, sodium
carboxymethylcellulose, a low melting wax, cocoa butter, and the
like. The term "preparation" is intended to include the formulation
of active compound with encapsulating material as a carrier
providing a capsule in which active component with or without other
carriers, is surrounded by a carrier, which is thus in association
with it. Similarly, cachets and lozenges are included within the
pharmaceutical formulations of the invention. Tablets, powders,
capsules, pills, cachets, and lozenges can be used as solid dosage
forms suitable for oral administration.
[0133] Suitable solid excipients are carbohydrate or protein
fillers and include, but are not limited to, sugars, including
lactose, sucrose, mannitol, or sorbitol; starch from corn, wheat,
rice, potato, or other plants; cellulose such as methyl cellulose,
hydroxypropylmethyl-cellulose, or sodium carboxymethylcellulose;
and gums including arabic and tragacanth; as well as proteins such
as gelatin and collagen. If desired, disintegrating or solubilizing
agents may be added, such as the cross-linked polyvinyl
pyrrolidone, agar, alginic acid, or a salt thereof, such as sodium
alginate.
[0134] Dragee cores are provided with suitable coatings such as
concentrated sugar solutions, which may also contain gum arabic,
talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol,
and/or titanium dioxide, lacquer solutions, and suitable organic
solvents or solvent mixtures. Dyestuffs or pigments may be added to
the tablets or dragee coatings for product identification or to
characterize the quantity of active compound (i.e., dosage).
Pharmaceutical preparations of the invention can also be used
orally using, for example, push-fit capsules made of gelatin, as
well as soft, sealed capsules made of gelatin and a coating such as
glycerol or sorbitol. Push-fit capsules can contain an
anti-inflammatory pyrazolopyrimidine mixed with a filler or binders
such as lactose or starches, lubricants such as talc or magnesium
stearate, and, optionally, stabilizers. In soft capsules, the
anti-inflammatory pyrazolopyrimidines may be dissolved or suspended
in suitable liquids, such as fatty oils, liquid paraffin, or liquid
polyethylene glycol with or without stabilizers.
[0135] For preparing suppositories, a low melting wax, such as a
mixture of fatty acid glycerides or cocoa butter, is first melted
and active component is dispersed homogeneously therein, as by
stirring. The molten homogeneous mixture is then poured into
convenient sized molds, allowed to cool, and thereby to
solidify.
[0136] Liquid form preparations include solutions, suspensions, and
emulsions, for example, water or water/propylene glycol solutions.
For parenteral injection, liquid preparations can be formulated in
solution in aqueous polyethylene glycol solution.
[0137] Aqueous solutions suitable for oral use can be prepared by
dissolving active component in water and adding suitable colorants,
flavors, stabilizers, and thickening agents as desired. Aqueous
suspensions suitable for oral use can be made by dispersing the
finely divided active component in water with viscous material,
such as natural or synthetic gums, resins, methylcellulose, sodium
carboxymethylcellulose, hydroxypropylmethylcellulose, sodium
alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia, and
dispersing or wetting agents such as a naturally occurring
phosphatide (e.g., lecithin), a condensation product of an alkylene
oxide with a fatty acid (e.g., polyoxyethylene stearate), a
condensation product of ethylene oxide with a long chain aliphatic
alcohol (e.g., heptadecaethylene oxycetanol), a condensation
product of ethylene oxide with a partial ester derived from a fatty
acid and a hexitol (e.g., polyoxyethylene sorbitol mono-oleate), or
a condensation product of ethylene oxide with a partial ester
derived from fatty acid and a hexitol anhydride (e.g.,
polyoxyethylene sorbitan mono-oleate). Aqueous suspension can also
contain one or more preservatives such as ethyl or n-propyl
p-hydroxybenzoate, one or more coloring agents, one or more
flavoring agents and one or more sweetening agents, such as
sucrose, aspartame or saccharin. Formulations can be adjusted for
osmolarity.
[0138] Also included are solid form preparations, which are
intended to be converted, shortly before use, to liquid form
preparations for oral administration. Such liquid forms include
solutions, suspensions, and emulsions. These preparations may
contain, in addition to active component, colorants, flavors,
stabilizers, buffers, artificial and natural sweeteners,
dispersants, thickeners, solubilizing agents, and the like.
[0139] Oil suspensions can be formulated by suspending an
anti-inflammatory pyrazolopyrimidine in a vegetable oil, such as
arachis oil, olive oil, sesame oil or coconut oil, or in a mineral
oil such as liquid paraffin; or a mixture of these. The oil
suspensions can contain a thickening agent, such as beeswax, hard
paraffin or cetyl alcohol. Sweetening agents can be added to
provide a palatable oral preparation, such as glycerol, sorbitol or
sucrose. These formulations can be preserved by addition of an
antioxidant such as ascorbic acid. As an example of an injectable
oil vehicle, see Minto, J Pharmacol. Exp. Ther. 281:93-102, 1997.
The pharmaceutical formulations of the invention can also be in the
form of oil-in-water emulsions. The oily phase can be a vegetable
oil or a mineral oil, described above, or a mixture of these.
Suitable emulsifying agents include naturally-occurring gums, such
as gum acacia and gum tragacanth, naturally occurring phosphatides,
such as soybean lecithin, esters or partial esters derived from
fatty acids and hexitol anhydrides, such as sorbitan mono-oleate,
and condensation products of these partial esters with ethylene
oxide, such as polyoxyethylene sorbitan mono-oleate. The emulsion
can also contain sweetening agents and flavoring agents, as in the
formulation of syrups and elixirs. Such formulations can also
contain a demulcent, a preservative, or a coloring agent.
[0140] The anti-inflammatory pyrazolopyrimidines of the invention
can be delivered transdermally, by a topical route, formulated as
applicator sticks, solutions, suspensions, emulsions, gels, creams,
ointments, pastes, jellies, paints, powders, and aerosols.
[0141] The anti-inflammatory pyrazolopyrimidines of the invention
can also be delivered as microspheres for slow release in the body.
For example, microspheres can be administered via intradermal
injection of drug-containing microspheres, which slowly release
subcutaneously (see Rao, J. Biomater Sci. Polym. Ed. 7:623-645,
1995; as biodegradable and injectable gel formulations (see, e.g.,
Gao Pharm. Res. 12:857-863, 1995); or, as microspheres for oral
administration (see, e.g., Eyles, J Pharm. Pharmacol. 49:669-674,
1997). Both transdermal and intradermal routes can in some
instances afford constant or near constant rates of delivery of the
active agents for weeks or months.
[0142] The anti-inflammatory pyrazolopyrimidine pharmaceutical
formulations of the invention can be provided as a salt and can be
formed with many acids, including but not limited to hydrochloric,
sulfuric, acetic, lactic, tartaric, malic, succinic, etc. Salts
tend to be more soluble in aqueous or other protonic solvents that
are the corresponding free base forms. In other cases, the
preparation may be a lyophilized powder in 1 mM-50 mM histidine,
0.1%-2% sucrose, 2%-7% mannitol at a pH range of 4.5 to 5.5, that
is combined with buffer prior to use.
[0143] In another embodiment, the anti-inflammatory
pyrazolopyrimidine formulations of the invention are used for
parenteral administration, such as intravenous (IV) administration
or administration into a body cavity or lumen of an organ. The
formulations for administration will commonly comprise a solution
of the anti-inflammatory pyrazolopyrimidine dissolved in a
pharmaceutically acceptable carrier. Among acceptable vehicles and
solvents that can be employed are water and Ringer's solution, an
isotonic sodium chloride. In addition, sterile fixed oils can
conventionally be 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 can likewise be used in the preparation of injectables.
These solutions are sterile and generally free of undesirable
matter. These formulations may be sterilized by conventional, well
known (for other compounds) sterilization techniques. The
formulations may contain pharmaceutically acceptable auxiliary
substances as required to approximate physiological conditions such
as pH adjusting and buffering agents, toxicity adjusting agents,
e.g., sodium acetate, sodium chloride, potassium chloride, calcium
chloride, sodium lactate and the like. The concentration of
anti-inflammatory pyrazolopyrimidine in these formulations can vary
widely, and will be selected primarily based on fluid volumes,
viscosities, body weight, and the like, in accordance with the
particular mode of administration selected and the patient's needs.
For IV administration, the formulation can be a sterile injectable
preparation, such as a sterile injectable aqueous or oleaginous
suspension. This suspension can be formulated according to the
known art using those suitable dispersing or wetting agents and
suspending agents. The sterile injectable preparation can also be a
sterile injectable solution or suspension in a nontoxic
parenterally-acceptable diluent or solvent, such as a solution of
1,3-butanediol.
[0144] In another embodiment, the anti-inflammatory
pyrazolopyrimidine formulations of the invention can be delivered
by the use of liposomes which fuse with the cellular membrane or
are endocytosed, i.e., by employing ligands attached to the
liposome, or attached directly to the oligonucleotide, that bind to
surface membrane protein receptors of the cell resulting in
endocytosis. By using liposomes, particularly where the liposome
surface carries ligands specific for target cells, or are otherwise
preferentially directed to a specific organ, one can focus the
delivery of the anti-inflammatory pyrazolopyrimidine into the
target cells in vivo. (See, e.g., Al-Muhammed, J Microencapsul.
13:293-306, 1996; Chonn, Curr. Opin. Biotechnol. 6:698-708, 1995;
Ostro, Am. J. Hosp. Pharm. 46:1576-1587, 1989).
[0145] The pharmaceutical preparation is preferably in unit dosage
form. In such form the preparation is subdivided into unit doses
containing appropriate quantities of active component. The unit
dosage form can be a packaged preparation, the package containing
discrete quantities of preparation, such as packeted tablets,
capsules, and powders in vials or ampoules. Also, the unit dosage
form can be a capsule, tablet, cachet, or lozenge itself, or it can
be appropriate number of any of these in packaged form.
[0146] The quantity of active component in a unit dose preparation
may be varied or adjusted from 0.1 mg to 10000 mg, more typically
1.0 mg to 1000 mg, most typically 10 mg to 500 mg, according to the
particular application and the potency of active component. The
composition can, if desired, also contain other compatible
therapeutic agents.
VII. Therapeutic Combinations
[0147] One of ordinary skill in art will appreciate that the
compounds of the invention are useful in treating a diverse array
of diseases. One of ordinary skill in art will also appreciate
that, when using the compounds of the invention in the treatment of
a specific disease, the compounds of the invention may be combined
or otherwise co-administered with various existing therapeutic
agents used for that disease. For example, for the treatment of
rheumatoid arthritis, the compounds of the invention may be
combined or co-administered with agents such as
TNF-.alpha.inhibitors such as anti-TNF monoclonal antibodies and
TNF receptor immunoglobulin molecules (such as Enbrel.R.TM.), low
dose methotrexate, lefunimide, hydroxychloroquine, d-penicilamine,
auranofin or parenteral or oral gold.
[0148] The compounds of the invention can also be used in
combination with existing therapeutic agents for the treatment of
osteoarthritis. Suitable agents to be used in combination include
standard non-steroidal anti-inflammatory agents (hereinafter
NSAIDs) such as piroxicam, diclofenac, propionic acids such as
naproxen, flurbiprofen, fenoprofen, ketoprofen and ibuprofen,
fenamates such as mefenamic acid, indomethacin, sulindac, apazone,
pyrazolones such as phenylbutazone, salicylates such as aspirin,
COX-2 inhibitors such as celecoxib and rofecoxib, analgesics and
intraarticular therapies such as corticosteroids and hyaluronic
acids such as hyalgan and synvisc.
[0149] Active ingredient (i.e. the compounds of Formula (I)) of the
present invention may be administered in combination with
inhibitors of other mediators of inflammation, comprising one or
more members selected from the group consisting essentially of the
classes of such inhibitors and examples thereof which include,
matrix metalloproteinase inhibitors, aggrecanase inhibitors, TACE
inhibitors, leukotriene receptor antagonists, IL-1 processing and
release inhibitors, IL-IRA, H.sub.1-receptor antagonists;
kinin-B.sub.1- and B.sub.1-receptor antagonists; prostaglandin
inhibitors such as PGD-, PGF- PGI.sub.2-, and PGE-receptor
antagonists; thromboxane A.sub.2(TXA2-) inhibitors; 5- and
12-lipoxygenase inhibitors; leukotriene LTC.sub.4-,
LTD.sub.4/LTE.sub.4-, and LTB.sub.4-inhibitors; PAF-receptor
antagonists; gold in the form of an aurothio group together with
various hydrophilic groups; immunosuppressive agents, e.g.,
cyclosporine, azathioprine, and methotrexate; anti-inflammatory
glucocorticoids; penicillamine; hydroxychloroquine; anti-gout
agents, e.g., colchicine, xanthine oxidase inhibitors, e.g.,
allopurinol, and uricosuric agents, e.g., probenecid,
sulfinpyrazone, and benzbromarone.
[0150] The compounds of the present invention may also be used in
combination with anticancer agents such as endostatin and
angiostatin or cytotoxic drugs such as adriamycin, daunomycin,
cis-platinum, etoposide, taxol, taxotere and alkaloids, such as
vincristine, and antimetabolites such as methotrexate.
[0151] The compounds of the present invention may also be used in
combination with anti-hypertensives and other cardiovascular drugs
intended to offset the consequences of atherosclerosis, including
hypertension, myocardial ischemia including angina, congestive
heart failure, and myocardial infarction, selected from
vasodilators such as hydralazine, .beta.-adrenergic receptor
antagonists such as propranolol, calcium channel blockers such as
nifedipine, .alpha..sub.2-adrenergic agonists such as clonidine,
.alpha.-adrenergic receptor antagonists such as prazosin, and
HMG-CoA-reductase inhibitors (anti-hypercholesterolemics) such as
lovastatin or atorvastatin.
[0152] Active ingredient of the present invention may also be
administered in combination with one or more antibiotic,
antifungal, antiprotozoal, antiviral or similar therapeutic
agents.
[0153] The compounds of the present invention may also be used in
combination with CNS agents such as antidepressants (such as
sertraline), anti-Parkinsonian drugs (such as L-dopa, requip,
mirapex, MAOB inhibitors such as selegine and rasagiline, comP
inhibitors such as Tasmar, A-2 inhibitors, dopamine reuptake
inhibitors, NMDA antagonists, nicotine agonists, dopamine agonists
and inhibitors of neuronal nitric oxide synthase), and
anti-Alzheimer's drugs such as donepezil, tacrine, COX-2
inhibitors, propentofylline or metrifonate.
[0154] The compounds of the present invention may also be used in
combination with osteoporosis agents such as roloxifene,
lasofoxifene, droloxifene or fosomax and immunosuppressant agents
such as FK-506 and rapamycin.
[0155] The present invention also relates to the formulation of
active agents of the present invention alone or with one or more
other therapeutic agents which are to form the intended
combination, including wherein said different drugs have varying
half-lives, by creating controlled-release forms of said drugs with
different release times which achieves relatively uniform dosing;
or, in the case of non-human patients, a medicated feed dosage form
in which said drugs used in the combination are present together in
admixture in said feed composition. There is further provided in
accordance with the present invention co-administration in which
the combination of drugs is achieved by the simultaneous
administration of said drugs to be given in combination; including
co-administration by means of different dosage forms and routes of
administration; the use of combinations in accordance with
different but regular and continuous dosing schedules whereby
desired plasma levels of said drugs involved are maintained in the
patient being treated, even though the individual drugs making up
said combination are not being administered to said patient
simultaneously.
[0156] This invention also relates to a method of or a
pharmaceutical composition for treating inflammatory processes and
diseases comprising administering a compound of this invention or
its salt to a mammal including a human, cat, livestock or dog
(preferably a dog), wherein said inflammatory processes and
diseases are defined as above, and said inhibitory compound is used
in combination with one or more other therapeutically active agents
under the following conditions: [0157] A.) where a joint has become
seriously inflamed as well as infected at the same time by
bacteria, fungi, protozoa, and/or virus, said inhibitory compound
is administered in combination with one or more antibiotic,
antifungal, antiprotozoal, and/or antiviral therapeutic agents;
[0158] B.) where a multi-fold treatment of pain and inflammation is
desired, said inhibitory compound is administered in combination
with inhibitors of other mediators of inflammation, comprising one
or more members independently selected from the group consisting
essentially of: [0159] (1) NSAID's; [0160] (2) H.sub.4-receptor
antagonists; [0161] (3) kinin-B.sub.4- and B.sub.4-receptor
antagonists; [0162] (4) prostaglandin inhibitors selected from the
group consisting of PGD-, PGF- PGI.sub.2-, and PGE-receptor
antagonists; [0163] (5) thromboxane A.sub.2 (TXA.sub.2-)
inhibitors; [0164] (6) 5-, 12- and 15-lipoxygenase inhibitors;
[0165] (7) leukotriene LTC.sub.4-, LTD.sub.4/LTE.sub.4-, and
LTB.sub.4-inhibitors; [0166] (8) PAF-receptor antagonists; [0167]
(9) gold in the form of an aurothio group together with one or more
hydrophilic groups; [0168] (10) immunosuppressive agents selected
from the group consisting of cyclosporine, azathioprine, and
methotrexate; [0169] (11) anti-inflammatory glucocorticoids; [0170]
(12) penicillamine; [0171] (13) hydroxychloroquine; [0172] (14)
anti-gout agents including colchicine; xanthine oxidase inhibitors
including allopurinol; and uricosuric agents selected from
probenecid, sulfinpyrazone, and benzbromarone; [0173] C.) where
older mammals are being treated for disease conditions, syndromes
and symptoms found in geriatric mammals, said inhibitory compound
is administered in combination with one or more members
independently selected from the group consisting essentially of:
[0174] (1) cognitive therapeutics to counteract memory loss and
impairment; [0175] (2) anti-hypertensives and other cardiovascular
drugs intended to offset the consequences of atherosclerosis,
hypertension, myocardial ischemia, angina, congestive heart
failure, and myocardial infarction, selected from the group
consisting of: [0176] a. diuretics; [0177] b. vasodilators; [0178]
c. .beta.-adrenergic receptor antagonists; [0179] d. angiotensin-II
converting enzyme inhibitors (ACE-inhibitors), alone or optionally
together with neutral endopeptidase inhibitors; [0180] e.
angiotensin II receptor antagonists; [0181] f. renin inhibitors;
[0182] g. calcium channel blockers; [0183] h. sympatholytic agents;
[0184] i. .alpha..sub.2-adrenergic agonists; [0185] j.
.alpha.-adrenergic receptor antagonists; and [0186] k.
HMG-CoA-reductase inhibitors (anti-hypercholesterolemics); [0187]
(3) antineoplastic agents selected from antimitotic drugs such as
the vinca alkaloids including but not limited to vinblastine and
vincristine; [0188] (4) growth hormone secretagogues; [0189] (5)
strong analgesics; [0190] (6) local and systemic anesthetics; and
[0191] (7) H.sub.2-receptor antagonists, proton pump inhibitors,
and other gastroprotective agents.
[0192] The term "treating", as used herein, refers to reversing,
alleviating, inhibiting the progress of, or preventing the disorder
or condition to which such term applies, or one or more symptoms of
such disorder or condition. The term "treatment", as used herein,
refers to act of treating, as "treating" is defined immediately
above.
[0193] The terms and expressions which have been employed herein
are used as terms of description and not of limitation, and there
is no intention in the use of such terms and expressions of
excluding equivalents of the features shown and described, or
portions thereof, it being recognized that various modifications
are possible within the scope of the invention claimed. Moreover,
any one or more features of any embodiment of the invention may be
combined with any one or more other features of any other
embodiment of the invention, without departing from the scope of
the invention. For example, the features of the anti-inflammatory
compounds of the present invention are equally applicable to the
methods of treating disease states and/or the pharmaceutical
compositions described herein. All publications, patents, and
patent applications cited herein are hereby incorporated by
reference in their entirety for all purposes.
EXAMPLES
Example 1
[0194] Various pyrazolopyrimidine compounds were tested for
anti-inflammatory properties using the methods detailed below.
[0195] Cytokines, Antibodies, and Reagents
[0196] Recombinant human interferon-.gamma. (IFN-.gamma.),
TNF-.alpha., interleukin (IL)-1.beta., and IL-4 were from R&D
Systems (Minneapolis, Minn.). Histamine was from Sigma (St. Louis,
Mo.). Mouse antibodies were obtained from commercial sources:
murine IgG and anti-human vascular endothelial growth factor
receptor-2 (VEGFR2) (mIgG1; Sigma), anti-human tissue factor
(mIgG1; Calbiochem, San Diego, Calif.), anti-human intercellular
adhesion molecule-1 (ICAM-1) (mIgG1; Beckman Coulter, Fullerton,
Calif.), and anti-human E-selectin (mIgG1; HyCult Biotechnology,
Uden, The Netherlands). Mouse antibodies against human vascular
cell adhesion molecule-1 (VCAM-1) (mIgG1), HLA-DR (mIgG2a), CD3
(mIgG1), CD40 (mIgG1), CD69 (mIgG1), MIG (mIgG1), MCP-1 (mIgG1),
CD14 (mIgG1), IL-1.alpha. (mIgG1), P-selectin (mIgG1), DAF
(mIgG2a), urokinase-type plasminogen activator receptor (uPAR)
(mIgG1), and CD38 (mIgG1) were obtained from BD Biosciences (San
Jose, Calif.). Mouse antibodies against eotaxin-3 (mIgG1), IL-8
(mIgG1), and M-CSF (mIgG1) were obtained from R&D Systems.
Staphylococcal enterotoxin B, toxic shock syndrome toxin-1
(staphylococcal enterotoxin F) from S. aureus (collectively called
superantigen; SAG), and lipopolysaccharide (LPS) from Salmonella
enteritidis were obtained from Sigma.
[0197] Human umbilical vein endothelial cells (HUVEC) were cultured
as described in Kunkel et al., 2004, FASEB J 18:1279-1281.
Peripheral blood mononuclear cells (PBMC) were prepared from buffy
coats (Stanford Blood Bank, Stanford, Calif.) by centrifugation
over Hisopaque-1077 (Sigma). Four assay systems, 3C, 4H, SAG, and
LPS, were used. For the 3C system, HUVEC were cultured for 24 h in
microtiter plates (Falcon; BD Biosciences), in the presence of
cytokines IL-1.beta. (1 ng/ml), TNF-.alpha. (5 ng/ml), and
IFN-.gamma. (20 ng/ml). For the 4H system, HUVEC were cultured in
the presence of IL-4 (5 ng/ml) and histamine (10 .mu.M). For the
SAG system, HUVEC were cultured with PBMC (7.5.times.10.sup.4) and
SAG (20 ng/ml). For the LPS system, HUVEC were cultured with PBMC
(7.5.times.10.sup.4) and LPS (2 ng/ml). Compounds were added 1 h
before stimulation and were present during the entire 24-h
stimulation period. Cell-based enzyme-linked immunosorbent assay
(ELISAs) were carried out as described. (See Kunkel et al., 2004,
FASEB J18:1279-1281.)
[0198] Data Analysis
[0199] Mean optical density values for each parameter measured by
ELISA were calculated from triplicate samples per experiment.
Well-to-well coefficients of variance range from 1 to 12%,
depending on the parameter measured, and average 5% across all
controls. Day-to-day variability for a given readout, system, and
treatment is the greatest contributor to the overall variability
(ranging from 10 to 60% of the total variability), but is
controlled for by using a prediction envelope to give the error
boundaries for all the measurements simultaneously, consistent with
our multivariate analysis approach. The envelope estimates the
variability of the measurements around the mean (all data are
centered). By combining similar measurements from multiple
experiments, overall error measures are established while
eliminating the specific bias of each experiment. Extensive studies
have been performed concerning the number of repeats required for
correctly classifying repeated profiles within given confidence
limits leading to the requirement for at least three replicate
wells per treatment and at least three independent repeats
(unpublished observations).
[0200] Function Similarity Maps
[0201] Within each experiment, mean optical density values were
used to generate ratios between treated (e.g., compound or siRNA)
and matched control (e.g., media or dimethyl sulfoxide) parameter
values. These normalized parameter ratios were then log 10
transformed. Log expression ratios were used in all Pearson
correlation calculations. Correlations were visualized in two
dimensions by multidimensional scaling using AT&T GraphViz
software. Distances between compounds are representative of their
similarities, and lines are drawn between compounds whose profiles
are similar at a level not due to chance. Significant correlations
were determined by (a) identifying the number of correlations that
exceed a given threshold in the observed Pearson correlation
distribution of the profiles, (b) calculating the average number of
Pearson correlations that exceed this threshold using correlations
calculated from randomized data made by permuting the empirical
profiles multiple times, (c) reselecting the Pearson correlation
threshold to minimize the false detection rate (FDR) (the FDR
provides the probability that a significant correlation is a false
positive), and (d) applying this cutoff Pearson correlation value
to the correlations between experimental profiles. This ensures
that for a 5% FDR, 95% of the correlations derived from the
experimental profiles are not due to chance.
[0202] The results are presented in Table 1 below. Structure 13 in
the Table is PP1 and not a compound of the invention. A "+" means
that a compound was not active in the assay at a concentration of 5
micromolar. In one embodiment, the compounds of the invention are
compounds other than compound nos. 1-57 in Table 1, below. In one
embodiment, the methods of the invention are practiced with a
compound shown in Table 1 other than PP1. In one embodiment, the
methods of the invention are practiced with a compound in Table 1
that is identified as "active" in the Activity column. In one
embodiment, the methods of the invention are practiced with a
compound other than compound nos. 1-57 in Table 1, below.
TABLE-US-00001 TABLE 1 No Compound Activity 1. ##STR00006## + 2.
##STR00007## + 3. ##STR00008## + 4. ##STR00009## + 5. ##STR00010##
+ 6. ##STR00011## active 7. ##STR00012## active 8. ##STR00013##
active 9. ##STR00014## active 10. ##STR00015## active 11.
##STR00016## active 12. ##STR00017## active 13. ##STR00018## active
14. ##STR00019## + 15. ##STR00020## active 16. ##STR00021## + 17.
##STR00022## active 18. ##STR00023## + 19. ##STR00024## active 20.
##STR00025## + 21. ##STR00026## + 22. ##STR00027## active 23.
##STR00028## + 24. ##STR00029## + 25. ##STR00030## + 26.
##STR00031## + 27. ##STR00032## + 28. ##STR00033## + 29.
##STR00034## + 30. ##STR00035## active 31. ##STR00036## active 32.
##STR00037## + 33. ##STR00038## + 34. ##STR00039## + 35.
##STR00040## + 36. ##STR00041## active 37. ##STR00042## + 38.
##STR00043## + 39. ##STR00044## active 40. ##STR00045## + 41.
##STR00046## active 42. ##STR00047## + 43. ##STR00048## + 44.
##STR00049## + 45. ##STR00050## + 46. ##STR00051## + 47.
##STR00052## active 48. ##STR00053## active 49. ##STR00054## + 50.
##STR00055## active 51. ##STR00056## + 52. ##STR00057## active 53.
##STR00058## active 54. ##STR00059## active 55. ##STR00060## + 56.
##STR00061## + 57. ##STR00062## + 58. ##STR00063## + 59.
##STR00064## active 60. ##STR00065## active 61. ##STR00066## + 62.
##STR00067## active 63. ##STR00068## active 64. ##STR00069## active
65. ##STR00070## active 66. ##STR00071## active 67. ##STR00072##
active 68. ##STR00073## + 69. ##STR00074## + 70. ##STR00075## + 71.
##STR00076## + 72. ##STR00077## active 73. ##STR00078## + 74.
##STR00079## + 75. ##STR00080## + 76. ##STR00081## active 77.
##STR00082## active 78. ##STR00083## active 79. ##STR00084## active
80. ##STR00085## active 81. ##STR00086## active 82. ##STR00087##
active 83. ##STR00088## active 84. ##STR00089## active 85.
##STR00090## active 86. ##STR00091## active 87. ##STR00092## active
88. ##STR00093## active 89. ##STR00094## active
[0203] In Table 1, the term "active" indicates that at least 3
parameters (in the 4 systems tested (3C, 4H, SAG, and LPS))
simultaneously showed activity falling outside a 99% prediction
envelope, as described above.
[0204] Table 2, below, shows certain compounds that did not exhibit
activity in the systems tested at a concentration of 10 micromolar.
In one embodiment, the compounds of the invention are compounds
other than the compounds shown in Table 2. In one embodiment, the
methods of the invention are practiced with a compound other than a
compound shown in Table 2, below.
TABLE-US-00002 TABLE 2 No Compound 1. ##STR00095## 2. ##STR00096##
3. ##STR00097## 4. ##STR00098## 5. ##STR00099## 6. ##STR00100##
Example 2
[0205] The following compounds provide illustrative
characterization data for certain pyrazolopyrimidine compounds. All
starting materials and synthetic reagents were purchased from
commercial suppliers unless otherwise noted. Acid chlorides that
were not readily commercially available were synthesized by
treating the corresponding carboxylic acids with excess oxalyl
chloride and catalytic DMF in diethyl ether, as described in Ward
and Rhee, 1991, Tetrahedron Lett. 32:7165-7166. Useful protocols
were adapted from Hanefeld et al. 1996, J. Chem. Soc., Perkin
Trans. 1, 1996, 1545-1552. One skilled in the art will immediately
recognize that any differences in naming convention is not intended
to limit the illustrative value of the examples disclosed
herein.
[0206]
1-tert-Butyl-3-(2,5-dimethylbenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-y-
lamine. White powder; .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta.
1.79 (s, 9H), 2.20 (s, 3H), 2.25 (s, 3H), 4.22 (s, 2H), 4.94 (s,
2H), 6.81 (s, 1H), 6.98 (d, 1H), 7.08 (d, 1H), 8.23 (s, 1H);
.sup.13C NMR (CDCl.sub.3, 100 MHz) .delta. 19.3, 21.0, 29.2, 33.1,
59.9, 100.9, 128.1, 129.1, 130.9, 133.6, 135.9, 136.3, 140.9,
154.3, 154.6, 157.7; HRMS (EI) molecular ion calculated for
C.sub.18H.sub.23N.sub.5 309.19535, found 309.19386.
[0207]
1-tert-Butyl-3-(3,5-dimethylbenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-y-
lamine. White powder; .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta.
1.80 (s, 9H), 2.25 (s, 6H), 4.20 (s, 2H), 5.00 (s, 2H), 6.80 (s,
2H), 6.88 (s, 1H), 8.22 (s, 1H); .sup.13C NMR (CDCl.sub.3, 100 MHz)
.delta. 21.3, 29.2, 35.1, 59.9, 100.7, 126.2, 129.0, 138.0, 138.9,
141.3, 154.4, 154.7, 157.7; HRMS (EI) molecular ion calculated for
C.sub.18H.sub.23N.sub.5 309.19535, found 309.19439.
[0208]
1-tert-Butyl-3-(3,4-dichlorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-y-
lamine. White powder; .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta.
1.79 (s, 9H), 4.24 (s, 2H), 5.07 (s, 2H), 7.02 (dd, 1H), 7.29 (d,
1H), 7.37 (d, 1H), 8.26 (s, 1H); .sup.13C NMR (CDCl.sub.3, 100 MHz)
.delta. 29.1, 34.3, 60.2, 100.6, 127.6, 130.2, 131.0, 131.4, 133.2,
138.4, 139.2, 154.6, 154.7, 157.5; HRMS (EI) molecular ion
calculated for C.sub.16H.sub.17N.sub.5Cl.sub.2 349.08610, found
349.08621.
[0209]
1-tert-Butyl-3-(4-chlorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylami-
ne. White powder; .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 1.77
(s, 9H), 4.25 (s, 2H), 5.16 (br s, 2H), 7.11 (d, J=8 Hz, 2H), 7.27
(d, J=8 Hz, 2H), 8.22 (s, 1H); .sup.13C NMR (CDCl.sub.3, 100 MHz)
.delta. 29.1, 34.5, 60.0, 100.6, 129.3, 129.6, 133.1, 136.6, 140.1,
154.5, 154.6, 157.6; HRMS (EI) molecular ion calculated for
C.sub.16H.sub.18N.sub.5Cl 315.12507, found 315.12545.
[0210]
1-tert-Butyl-3-(4-methoxybenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylam-
ine. White powder; .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 1.78
(s, 9H), 3.76 (s, 3H), 4.22 (s, 2H), 4.91 (br s, 2H), 6.84 (d, J=9
Hz, 2H), 7.11 (d, J=9 Hz, 2H), 8.22 (s, 1H); .sup.13C NMR
(CDCl.sub.3, 100 MHz) .delta. 29.2, 34.3, 55.2, 59.9, 100.6, 114.6,
129.4, 130.0, 141.4, 154.5, 154.7, 157.6, 158.7; HRMS (EI)
molecular ion calculated for C.sub.17H.sub.21N.sub.5O 311.17461,
found 311.17454.
[0211]
1-tert-Butyl-3-(2-methoxybenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylam-
ine. White powder; .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 1.77
(s, 9H), 3.89 (s, 3H), 4.25 (s, 2H), 5.77 (br s, 2H), 6.89 (m, 2H),
7.18 (m, 2H), 8.20 (s, 1H); .sup.13C NMR (CDCl.sub.3, 100 MHz)
.delta. 27.4, 29.2, 55.5, 59.8, 100.4, 110.8, 121.5, 126.5, 128.0,
130.6, 141.1, 154.3, 154.3, 155.7, 158.0; HRMS.
[0212]
1-tert-Butyl-3-(2-chlorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylami-
ne. White powder; .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 1.78
(s, 9H), 4.38 (s, 2H), 5.41 (br s, 2H), 7.00 (dd, J.sub.1=7 Hz,
J.sub.2=2 Hz, 1H), 7.15 (m, 2H), 7.39 (dd, J.sub.1=8 Hz, J.sub.2=2
Hz, 1H), 8.22 (s, 1H); .sup.13C NMR (CDCl.sub.3, 100 MHz) .delta.
29.1, 31.9, 60.0, 100.7, 127.4, 128.4, 129.6, 130.1, 133.3, 135.6,
139.4, 154.4, 154.5, 157.7; HRMS (EI) molecular ion calculated for
C.sub.16H.sub.18N.sub.5Cl 315.12507, found 315.12449.
[0213]
1-tert-Butyl-3-(2-methylbenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylami-
ne. White powder; .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 1.77
(s, 9H), 2.30 (s, 3H), 4.25 (s, 2H), 5.09 (br s, 2H), 6.98 (d, J=8
Hz, 1H), 7.14 (m, 3H), 8.20 (s, 1H); .sup.13C NMR (CDCl.sub.3, 100
MHz) .delta. 19.8, 29.1, 33.1, 59.9, 100.9, 126.7, 127.3, 128.3,
130.8, 136.1, 136.8, 140.6, 154.3, 154.5, 157.8; HRMS (EI)
molecular ion calculated for C.sub.17H.sub.21N.sub.5 295.17970,
found 295.17922.
[0214]
1-tert-Butyl-3-(4-methylbenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylami-
ne. White powder; .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 1.79
(s, 9H), 2.30 (s, 3H), 4.24 (s, 2H), 4.94 (br s, 2H), 7.08 (d, J=8
Hz, 2H), 7.11 (d, J=8 Hz, 2H), 8.21 (s, 1H); .sup.13C NMR
(CDCl.sub.3, 100 MHz) .delta. 21.0, 29.2, 34.8, 59.9, 100.6, 128.3,
130.0, 135.1, 136.9, 141.3, 154.5, 154.7, 157.7; HRMS (EI)
molecular ion calculated for C.sub.17H.sub.21N.sub.5 295.17970,
found 295.18068.
[0215]
1-tert-Butyl-3-cyclopentylmethyl-1H-pyrazolo[3,4-d]pyrimidin-4-ylam-
ine. White powder; .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 1.28
(m, 2H), 1.53 (m, 2H), 1.65 (m, 2H), 1.71 (m, 2H), 1.73 (s, 9H),
2.28 (m, 1H), 2.86 (d, J=8 Hz, 2H), 5.69 (br s, 2H), 8.25 (s, 1H);
.sup.13C NMR (CDCl.sub.3, 100 MHz) .delta. 25.0, 29.2, 32.4, 35.2,
39.7, 59.7, 100.5, 142.1, 153.0, 154.0, 158.0; HRMS (EI) molecular
ion calculated for C.sub.15H.sub.23N.sub.5 273.19535, found
273.19565.
[0216]
4-Amino-1-tert-butyl-3-(1'-naphthyl)pyrazolo[3,4-d]pyrimidine:
white powder; .sup.1H NMR (270 MHz, CDCl.sub.3) .delta. 1.92 (s,
9H), 5.04 (m, 2H), 7.43-7.73 (m, 4H), 7.92-8.02 (m, 3H), 8.34 (s,
1H); HRMS (EI) molecular ion calculated for C.sub.19H.sub.19N.sub.5
317.16427, found 317.16247.
[0217]
4-Amino-1-tert-butyl-3-(2'-naphthyl)pyrazolo[3,4-d]pyrimidine:
white powder; .sup.1H NMR (270 MHz, CDCl.sub.3) .delta. 1.88 (s,
9H), 5.55 (m, 2H), 7.56-8.00 (m, 6H), 8.16 (s, 1H), 8.39 (s, 1H);
HRMS (EI) molecular ion calculated for C.sub.19H.sub.19N.sub.5
317.16427, found 317.16359.
[0218]
4-Amino-1-tert-butyl-3-(m-phenoxyphenyl)pyrazolo[3,4-d]pyrimidine:
white powder; .sup.1H NMR (270 MHz, CDCl.sub.3) .delta. 1.83 (s,
9H), 5.61 (s, 2H), 7.08-7.49 (m, 9H), 8.35 (s, 1H); HRMS (EI)
molecular ion calculated for C.sub.21H.sub.21N.sub.5O 359.17483,
found 359.17325.
[0219]
4-Amino-1-tert-butyl-3-(m-benzyloxyphenyl)pyrazolo[3,4-d]pyrimidine-
: white powder; .sup.1H NMR (270 MHz, CDCl.sub.3) .delta. 1.85 (s,
9H), 5.17 (s, 2H), 5.55 (s, 2H), 7.10 (d, J) 8 Hz, 1H), 7.27-7.48
(m, 8H), 8.34 (s, 1H); HRMS (EI) molecular ion calculated for
C.sub.22H.sub.23N.sub.5O 373.19049, found 373.18833.
[0220]
4-Amino-1-tert-butyl-3-(m-(2',6'-dichloro)benzyloxyphenyl)pyrazolo[-
3,4-d]pyrimidine: white powder; .sup.1H NMR (270 MHz, CDCl.sub.3)
.delta. 1.85 (s, 9H), 5.36 (s, 2H), 5.74 (s, 2H), 7.11-7.51 (m,
7H), 8.36 (s, 1H); HRMS (EI) molecular ion calculated for
C.sub.22H.sub.21Cl.sub.2N.sub.5O 441.11263, found 441.11050.
[0221] 4-Amino-1-tert-butyl-3-piperonylpyrazolo[3,4-d]pyrimidine:
white powder; .sup.1H NMR (270 MHz, CDCl.sub.3) .delta. 1.83 (s,
9H), 5.70 (s, 2H), 6.05 (s, 2H), 6.96 (d, J) 8 Hz, 1H), 7.13-7.27
(m, 2H), 8.34 (s, 1H); HRMS (EI) molecular ion calculated for
C.sub.16H.sub.17N.sub.5O.sub.2 311.13841, found 311.13777.
[0222]
4-Amino-1-tert-butyl-3-(p-tert-butylphenyl)pyrazolo[3,4-d]pyrimidin-
e: white powder; .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 1.38 (s,
9H), 1.84 (s, 9H), 5.83 (s, 2H), 7.58 (dd, J) 8 Hz, 12 Hz, 4H),
8.33 (s, 1H); HRMS (EI) molecular ion calculated for
C.sub.19H.sub.25N.sub.5 323.21125, found 323.21024.
[0223]
4-Amino-1-tert-butyl-3-(1'-naphthylmethyl)pyrazolo[3,4-d]pyrimidine-
: white powder; .sup.1H NMR (270 MHz, CDCl.sub.3) .delta. 1.85 (s,
9H), 4.76 (s, 2H), 5.04 (s, 2H), 7.19 (d, J) 6 Hz, 1H), 7.39 (t, J)
8 Hz, 1H), 7.55 (t, J) 4 Hz, 2H), 7.79-7.92 (m, 2H), 8.20 (d, J) 8
Hz, 1H), 8.24 (s, 1H); HRMS (EI) molecular ion calculated for
C.sub.20H.sub.21N.sub.5 331.17993, found 331.17951.
[0224]
4-Amino-1-tert-butyl-3-(1'-naphthoxymethyl)pyrazolo[3,4-d]pyrimidin-
e: beige powder; .sup.1H NMR (270 MHz, CDCl.sub.3) .delta. 1.83 (s,
9H), 5.57 (s, 2H), 6.12 (s, 2H), 7.07 (d, J) 7 Hz, 1H), 7.39-7.54
(m, 4H), 7.84 (d, J) 8 Hz, 1H), 8.25 (d, J) 8 Hz, 1H), 8.35 (s,
1H); HRMS (EI) molecular ion calculated for
C.sub.20H.sub.21N.sub.5O 347.17483, found 347.17408.
Example 3
[0225] The following example illustrates the ability of a compound
useful in the methods of the present invention to reduce neutrophil
and monocyte recruitment, as well as to reduce the total number of
leukocytes in a mouse peritoneal inflammation model.
[0226] A single dose (10 mg/kg in 200 .mu.l) of an
anti-inflammatory pyrazolopyrimidine compound of the present
invention (compound 30 in Table 1, above) was administered
intraperionteally to a group of 6 mice while another group of 6
mice received an intraperitoneal injection of vehicle only. Fifteen
minutes later, an inflammatory stimulus (thioglycollate, 200 .mu.l)
was administered intraperitoneally to each mouse. Peritoneal lavage
was collected at 4 and 24 hours subsequent to stimulus
administration.
[0227] Inflammatory infiltrate was recovered by injection of 10 ml
of Ca++/Mg++free PBS, massaging, and withdrawal. Total recovered
cells were counted on a hemacytometer. The results are presented in
FIG. 2 (average+/-SD of 6 mice).
[0228] Neutrophils, monocytes, lymophocytes and eosinophils were
enumerated by analyzing smears of the peritoneal wash stained with
Wright-Geimsa stain. The results are presented in FIG. 3
(average+/-SD of 6 mice).
* * * * *