U.S. patent application number 10/183179 was filed with the patent office on 2003-07-24 for n-heterocyclic inhibitors of tnf-alpha expression.
Invention is credited to Ahmed, Gulzar, Diller, David J., Henderson, Ian, Leftheris, Katerina, Metzger, Axel, Wen, James, Wrobleski, Stephen T..
Application Number | 20030139435 10/183179 |
Document ID | / |
Family ID | 23161582 |
Filed Date | 2003-07-24 |
United States Patent
Application |
20030139435 |
Kind Code |
A1 |
Ahmed, Gulzar ; et
al. |
July 24, 2003 |
N-heterocyclic inhibitors of TNF-alpha expression
Abstract
N-heterocyclic compounds that block cytokine production via
inhibition of p38 kinase are disclosed. In one embodiment,
compounds of the present invention are represented by Formula (I):
1 Methods of production, pharmaceutical compositions and methods of
treating conditions associated with inappropriate p38 kinase
activity or TNF-.alpha. expression utilizing compounds of the
present invention are also disclosed.
Inventors: |
Ahmed, Gulzar; (Yardley,
PA) ; Metzger, Axel; (East Windsor, NJ) ;
Henderson, Ian; (Hopewell, NJ) ; Diller, David
J.; (Hightstown, NJ) ; Wen, James; (Dayton,
NJ) ; Wrobleski, Stephen T.; (Whitehouse Station,
NJ) ; Leftheris, Katerina; (Skillman, NJ) |
Correspondence
Address: |
STEPHEN B. DAVIS
BRISTOL-MYERS SQUIBB COMPANY
PATENT DEPARTMENT
P O BOX 4000
PRINCETON
NJ
08543-4000
US
|
Family ID: |
23161582 |
Appl. No.: |
10/183179 |
Filed: |
June 26, 2002 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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60301020 |
Jun 26, 2001 |
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Current U.S.
Class: |
514/275 ;
514/336; 514/352; 544/330; 544/331; 546/268.1; 546/308 |
Current CPC
Class: |
A61P 17/02 20180101;
A61P 19/06 20180101; A61P 9/04 20180101; A61P 19/02 20180101; C07D
403/12 20130101; A61P 9/00 20180101; A61P 7/04 20180101; A61P 9/10
20180101; C07D 239/50 20130101; A61P 37/02 20180101; A61P 1/18
20180101; A61P 25/16 20180101; A61P 7/06 20180101; C07D 213/85
20130101; A61P 37/04 20180101; A61P 17/06 20180101; A61P 1/04
20180101; A61P 7/02 20180101; C07D 239/48 20130101; A61P 35/04
20180101; A61P 35/00 20180101; A61P 25/28 20180101; A61P 13/12
20180101; A61P 25/02 20180101; A61P 3/10 20180101; A61P 31/18
20180101; A61P 37/06 20180101; A61P 37/08 20180101; A61P 25/00
20180101; A61P 31/00 20180101; A61P 29/00 20180101; A61P 27/02
20180101; A61P 1/02 20180101; A61P 1/16 20180101; A61P 29/02
20180101; A61P 21/04 20180101; A61P 19/10 20180101; A61P 31/12
20180101; A61P 19/08 20180101; C07D 239/47 20130101; A61P 17/00
20180101; A61P 43/00 20180101; C07D 401/12 20130101 |
Class at
Publication: |
514/275 ;
514/352; 514/336; 544/330; 544/331; 546/268.1; 546/308 |
International
Class: |
A61K 031/506; A61K
031/505; C07D 41/02; C07D 43/02; A61K 031/4439 |
Claims
What is claimed is:
1. A compound of Formula (I), 182including isomers, enantiomers,
diastereomers, tautomers, pharmaceutically acceptable salts,
prodrugs and solvates thereof, wherein: one or two of W, Y and X
are .dbd.N--; one of W, Y and X is selected from .dbd.C--CN,
.dbd.C--F, .dbd.C--NO.sub.2, .dbd.C--Br, .dbd.C--NH.sub.2,
.dbd.C--NHC(O)CH.sub.3 and .dbd.C--Cl; the remaining W, Y or X is
.dbd.CH--; V is --NR.sup.5--; Z is halogen or
--N(R.sup.1)(R.sup.2); R.sup.1 and R.sup.2 are the same or
different and are selected from hydrogen, alkyl, substituted alkyl,
aryl, substituted aryl, cycloalkyl, substituted cycloalkyl,
heterocyclyl or substituted heterocyclyl; R.sup.5 is hydrogen or
alkyl; R.sup.6 is 183R.sup.7 is hydrogen, alkyl, substituted alkyl,
alkoxy, or halogen; R.sup.8 is hydrogen, alkyl, alkyloxy or cyano;
R.sup.9 is --C(O)R.sup.10 or unsubstituted or substituted
heterocyclyl; R.sup.10 is --N(R.sup.31)(R.sup.32); R.sup.31 and
R.sup.32 are the same or different and are selected from hydrogen,
alkyl, substituted alkyl, alkoxy, aryl, substituted aryl,
cycloalkyl, substituted cycloalkyl, heterocyclyl or substituted
heterocyclyl; R.sup.11 is hydrogen, halogen, O--R.sup.35 or
--N(R.sup.12)(R.sup.13); R.sup.12 is hydrogen, alkyl, or
substituted alkyl; R.sup.13 is --(CH.sub.2).sub.mR.sup.14;
--N(R.sup.12)(R.sup.13) taken together may form a heterocyclyl or
substituted heterocyclyl; m is 0, 1, 2 or 3; R.sup.14 is hydrogen,
alkyl, substituted alkyl, --C(O)N(R.sup.31)(R.sup.32),
--N(R.sup.33)C(O)R.sup.34, aryl, substituted aryl, cycloalkyl,
substituted cycloalkyl, heterocyclyl, substituted heterocyclyl,
heteroaryl, substituted heteroaryl or 184R.sup.15 is hydrogen,
alkyl or substituted alkyl; R.sup.16 is hydrogen or alkyl; or
R.sup.33 is hydrogen, alkyl, or substituted alkyl; R.sup.34 is
alkyl, substituted alkyl, aryl or substituted aryl; R.sup.35 is
hydrogen or -(lower alkyl)-R.sup.36; R.sup.36 is
N(R.sup.37)(R.sup.38); R.sup.37 is hydrogen, alkyl, or substituted
alkyl; R.sup.38 is -(substituted alkyl)-R.sup.14; and
N(R.sup.37)(R.sup.38) taken together may form a heterocyclyl or
substituted heterocyclyl.
2. A compound of claim 1, including isomers, enantiomers,
diastereomers, tautomers, pharmaceutically acceptable salts,
prodrugs and solvates thereof, wherein: one or two of W, Y and X
are .dbd.N--; one of W, Y and X is selected from .dbd.C--CN,
.dbd.C--F, .dbd.C--NO.sub.2, .dbd.C--Br, .dbd.C--NH.sub.2,
.dbd.C--NHC(O)CH.sub.3 and .dbd.C--Cl; the remaining W, Y or X is
.dbd.CH--; V is --NH--; Z is --N(R.sup.1)(R.sup.2); R.sup.1 and
R.sup.2 are the same or different and are selected from hydrogen,
alkyl or substituted alkyl wherein alkyl is of 1 to 8 carbons;
R.sup.6 is 185R.sup.7 is hydrogen, alkyl of 1 to 4 carbons, alkoxy
of 1 to 4 carbons, or halogen; R.sup.8 is hydrogen; R.sup.9 is
--C(O)R.sup.10 or unsubstituted or substituted heterocyclyl;
R.sup.10 is --NH.sub.2 or unsubstituted or substituted --NH-alkyl,
--NH-alkoxy, --NH-heterocyclyl, --NH-phenyl, or
--NH-CH.sub.2-phenyl wherein alkyl and alkoxy are of 1 to 6
carbons; R.sup.11 is hydrogen, halogen, O--R.sup.35 or
--N(R.sup.12)(R.sup.13), wherein N(R.sup.12)(R.sup.13) taken
together may form a monocyclic heterocyclyl or substituted
heterocyclyl of 5 to 7 atoms containing 1, 2, or 3 additional
nitrogen atoms or wherein R.sup.12 is hydrogen; R.sup.13 is alkyl
of 1 to 4 carbons or 186R.sup.15 and R.sup.16 are independently
selected from hydrogen and methyl; R.sup.35 is hydrogen or -(lower
alkyl)-R.sup.36; R.sup.36 is N(R.sup.37)(R.sup.38); R.sup.37 is
hydrogen, alkyl, or substituted alkyl; R.sup.38 is -(substituted
alkyl)-R.sup.14; and N(R.sup.37)(R.sup.38) taken together may form
a heterocyclyl or substituted heterocyclyl.
3. A compound of claim 2, including isomers, enantiomers,
diastereomers, tautomers, pharmaceutically acceptable salts,
prodrugs and solvates thereof, wherein: one or two of W, Y and X
are .dbd.N--; one of W, Y and X is selected from .dbd.C--CN,
.dbd.C--F, .dbd.C--NO.sub.2, .dbd.C--Br, .dbd.C--NH.sub.2,
.dbd.C--NHC(O)CH.sub.3 and .dbd.C--Cl; the remaining W, Y or X is
.dbd.CH--; V is --NH--; Z is --N(R.sup.1)(R.sup.2); R.sup.1 and
R.sup.2 are the same or different and are selected from hydrogen or
alkyl of 1 to 8 carbons; R.sup.6 is 187R.sup.7 is hydrogen, methyl,
methoxy, Cl, Br, or F; R.sup.8 is hydrogen; R.sup.9 is
--C(O)R.sup.10 or unsubstituted or substituted heterocyclyl;
R.sup.10 is --NH.sub.2, or unsubstituted or substituted --NH-alkyl,
--NH-alkoxy, --NH-phenyl, or --NH--CH.sub.2-phenyl wherein alkyl
and alkoxy are of 1 to 6 carbons; and R.sup.11 is hydrogen,
halogen, O--R.sup.35 or --N(R.sup.12)(R.sup.13), wherein
N(R.sup.12)(R.sup.13) taken together form a monocyclic heterocyclyl
or substituted heterocyclyl of 5 to 7 atoms containing 1, 2, or 3
additional nitrogen atoms.
4. A compound of claim 3, including isomers, enantiomers,
diastereomers, tautomers, pharmaceutically acceptable salts,
prodrugs and solvates thereof, wherein: one of W, Y and X is
.dbd.N--; one of W, Y and X is selected from .dbd.C--CN, .dbd.C--F,
.dbd.C--NO.sub.2, .dbd.C--Br, .dbd.C--NH.sub.2,
.dbd.C--NHC(O)CH.sub.3 and .dbd.C--Cl; the remaining W, Y or X is
.dbd.CH--; V is --NH--; Z is --N(R.sup.1)(R.sup.2); R.sup.1 and
R.sup.2 are the same or different and are selected from hydrogen or
alkyl of 1 to 8 carbons; R.sup.6 is 188R.sup.7 is hydrogen, methyl,
methoxy, Cl, Br, or F; R.sup.8 is hydrogen; R.sup.9 is
--C(O)R.sub.10 or unsubstituted or substituted heterocyclyl;
R.sup.10 is --NH.sub.2, or unsubstituted or substituted --NH-alkyl,
--NH-alkoxy, --NH-phenyl, or --NH--CH.sub.2-phenyl wherein alkyl
and alkoxy are of 1 to 6 carbons; R.sup.11 is hydrogen, halogen,
--O--R.sup.35 or --N(R.sup.12)(R.sup.13), wherein
N(R.sup.12)(R.sup.13) taken together form a monocyclic heterocyclyl
or substituted heterocyclyl of 5 to 7 atoms containing 1, 2, or 3
additional nitrogen atoms; and R.sup.15 and R.sup.16 are
independently selected from hydrogen and methyl.
5. A compound of claim 4, including isomers, enantiomers,
diastereomers, tautomers, pharmaceutically acceptable salts,
prodrugs and solvates thereof, wherein: R.sup.10 is --NH.sub.2,
unsubstituted or substituted --NH--CH.sub.3, --NH--C.sub.2H.sub.5,
--NH--OCH.sub.3, or --NH--OC.sub.2H.sub.5.
6. A compound of claim 4, including isomers, enantiomers,
diastereomers, tautomers, pharmaceutically acceptable salts,
prodrugs and solvates thereof, wherein: R.sup.9 is unsubstituted or
substituted triazole, thiazole, oxadiazole or imidazole.
7. A compound of claim 5, including isomers, enantiomers,
diastereomers, tautomers, pharmaceutically acceptable salts,
prodrugs and solvates thereof, wherein: R.sup.11 is hydrogen,
halogen, --O-(substituted alkyl), --NH-(substituted alkyl) or
189
8. A compound of claim 6, including isomers, enantiomers,
diastereomers, tautomers, pharmaceutically acceptable salts,
prodrugs and solvates thereof, wherein: R.sup.11 is hydrogen,
halogen, --O-(substituted alkyl), --NH-(substituted alkyl) or
190
9. A compound of claim 3, including isomers, enantiomers,
diastereomers, tautomers, pharmaceutically acceptable salts,
prodrugs and solvates thereof, wherein: two of W, Y and X are
.dbd.N--; the remaining W, Y or X is selected from .dbd.C--CN,
.dbd.C--F, .dbd.C--NO.sub.2, .dbd.C--Br, .dbd.C--NH.sub.2,
.dbd.C--NHC(O)CH.sub.3, and .dbd.C--Cl; V is --NH--; Z is
--N(R.sup.1)(R.sup.2); R.sup.1 and R.sup.2 are the same or
different and are selected from hydrogen or alkyl of 1 to 8
carbons; R.sup.6 is 191R.sup.7 is hydrogen, methyl, methoxy, Cl,
Br, or F; R.sup.8 is hydrogen; R.sup.9 is --C(O)R.sup.10 or
unsubstituted or substituted heterocyclyl; R.sup.10 is --NH.sub.2,
or unsubstituted or substituted --NH-alkyl, --NH-alkoxy,
--NH-phenyl, or --NH--CH.sub.2-phenyl wherein alkyl and alkoxy are
of 1 to 6 carbons; R.sup.11 is hydrogen, halogen, --O--R.sup.35 or
--N(R.sup.12)(R.sup.13), wherein N(R.sup.12)(R.sup.13) taken
together may form a monocyclic heterocyclyl or substituted
heterocyclyl of 5 to 7 atoms containing 1, 2, or 3 additional
nitrogen atoms; and R.sup.15 and R.sup.16 are independently
selected from hydrogen and methyl.
10. A compound of claim 9, including isomers, enantiomers,
diastereomers, tautomers, pharmaceutically acceptable salts,
prodrugs and solvates thereof, wherein: R.sup.10 is --NH.sub.2,
unsubstituted or substituted --NH--CH.sub.3, --NH--C.sub.2H.sub.5,
--NH--OCH.sub.3, or --NH--OC.sub.2H.sub.5.
11. A compound of claim 9, including isomers, enantiomers,
diastereomers, tautomers, pharmaceutically acceptable salts,
prodrugs and solvates thereof, wherein: R.sup.9 is unsubstituted or
substituted triazole, thiazole, oxadiazole or imidazole.
12. A compound of claim 10, including isomers, enantiomers,
diastereomers, tautomers, pharmaceutically acceptable salts,
prodrugs and solvates thereof, wherein: R.sup.11 is hydrogen,
halogen, --O-(substituted alkyl), --NH-(substituted alkyl) or
192
13. A compound of claim 11, including isomers, enantiomers,
diastereomers, tautomers, pharmaceutically acceptable salts,
prodrugs and solvates thereof, wherein: R.sup.11 is hydrogen,
halogen, --O-(substituted alkyl), --NH-(substituted alkyl) or
193
14. A pharmaceutical composition comprising as an active
ingredient, a compound, or a prodrug or salt thereof, according to
claim 1, and a pharmaceutically acceptable carrier.
15. A pharmaceutical composition according to claim 14, further
comprising one or more additional active ingredients.
16. A pharmaceutical composition according to claim 15, wherein
said additional active ingredient is an anti-inflammatory
compound.
17. A pharmaceutical composition according to claim 16, wherein
said additional active ingredient is chosen from a steroid and an
NSAID.
18. A method of inhibiting TNF-.alpha. expression in a mammal, the
method comprising administering to the mammal an effective amount
of a composition according to claim 14.
19. A method of treating TNF-.alpha. mediated disorder, the method
comprising administering to a mammal in need of such treatment, an
effective amount of a composition according to claim 14.
20. The method according to claim 19, wherein the TNF-.alpha.
mediated disorder is an inflammatory disorder.
21. The method according to claim 19, wherein the TNF-.alpha.
mediated disorder is chosen from bone resorption, graft vs. host
reaction, atherosclerosis, arthritis, osteoarthritis, rheumatoid
arthritis, gout, psoriasis, topical inflammatory disease states,
adult respiratory distress syndrome, asthma, chronic pulmonary
inflammatory disease, cardiac reperfusion injury, renal reperfusion
injury, thrombus, glomerulonephritis, Chron's disease, ulcerative
colitis, inflammatory bowel disease, multiple sclerosis, endotoxin
shock, osteoporosis, Alzheimer's disease, congestive heart failure
and cachexia.
22. The method according to claim 19, wherein said composition
according to claim 16 is administered with one or more additional
anti-inflammatory or immunosuppressive agents as a single dose form
or as separate dosage forms.
23. A method of treating a condition associated with TNF-.alpha.
expression in a mammal, the method comprising administering to a
mammal in need of such treatment, an effective amount of a
composition according to claim 14.
24. The method according to claim 23, wherein the condition
associated with TNF-.alpha. expression is an inflammatory
disorder.
25. The method according to claim 23, wherein the condition
associated with TNF-.alpha. expression is chosen from bone
resorption, graft vs. host reaction, atherosclerosis, arthritis,
osteoarthritis, rheumatoid arthritis, gout, psoriasis, topical
inflammatory disease states, adult respiratory distress syndrome,
asthma, chronic pulmonary inflammatory disease, cardiac reperfusion
injury, renal reperfusion injury, thrombus, glomerulonephritis,
Crohn's disease, ulcerative colitis, inflammatory bowel disease,
multiple sclerosis, endotoxin shock, osteoporosis, Alzheimer's
disease, congestive heart failure and cachexia.
26. The method according to claim 23 wherein said composition
according to claim 16 is administered with one or more additional
anti-inflammatory or immunosupressive agents as a single dose form
or as separate dosage forms.
27. A method of treating a condition associated with p38 kinase
activity in a mammal, the method comprising administering to a
mammal in need of such treatment, an effective amount of a
composition according to claim 14.
28. The method according to claim 27, wherein the condition
associated with p38 kinase activity is an inflammatory
disorder.
29. The method according to claim 27, wherein the condition
associated with p38 kinase activity is chosen from bone resorption,
graft vs. host reaction, atherosclerosis, arthritis,
osteoarthritis, rheumatoid arthritis, gout, psoriasis, topical
inflammatory disease states, adult respiratory distress syndrome,
asthma, chronic pulmonary inflammatory disease, cardiac reperfusion
injury, renal reperfusion injury, thrombus, glomerulonephritis,
Crohn's disease, ulcerative colitis, inflammatory bowel disease,
multiple sclerosis, endotoxin shock, osteoporosis, Alzheimer's
disease, congestive heart failure and cachexia.
30. The method according to claim 27 wherein said composition
according to claim 14 is administered with one or more additional
anti-inflammatory or immunosuppressive agents as a single dose form
or as separate dosage forms.
31. A compound of claim 1, including isomers, enantiomers,
diastereomers, tautomers, pharmaceutically acceptable salts,
prodrugs and solvates thereof, wherein said compound is selected
from: 19419519619719819920020- 1202203204205206207208209210211212
Description
[0001] This application claims priority from provisional U.S.
Patent Application Serial No. 60/301,020, filed Jun. 26, 2001,
which is incorporated herein by reference in its entirety.
FIELD OF THE INVENTION
[0002] This invention relates to N-heterocyclic compounds that are
effective in blocking cytokine production, and in particular the
expression of TNF-alpha (TNF-.alpha.), via inhibition of p38
kinase. Compounds of the present invention are useful in the
treatment of inflammatory diseases such as, for example, rheumatoid
arthritis.
BACKGROUND OF THE INVENTION
[0003] Overproduction of cytokines such as IL-1 and TNF-.alpha. is
implicated in a wide variety of inflammatory diseases, including
rheumatoid arthritis (RA), psoriasis, multiple sclerosis,
inflammatory bowel disease, endotoxin shock, osteoporosis,
Alzheimer's disease and congestive heart failure, among others
[Henry et al., Drugs Fut., 24:1345-1354 (1999); Salituro et al.,
Curr. Med. Chem., 6:807-823 (1999)]. There is convincing evidence
in human patients that protein antagonists of cytokines, such as,
for example, monoclonal antibody to TNF-.alpha. (Enbrel) [Rankin et
al., Br. J. Rheumatol., 34:334-342 (1995)], soluble TNF-.alpha.
receptor-Fc fusion protein (Etanercept) [Moreland et al., Ann.
Intern. Med., 130:478-486 (1999)] and or IL-1 receptor antagonist
[Bresnihan et al., Arthritis Rheum., 41:2196-2204 (1998)], can
provide effective treatment for chronic inflammatory diseases. As
none of the current treatments for inflammatory diseases provide
complete relief of symptoms, and as most current treatments are
associated with various drawbacks such as side effects, improved
methods for treating inflammatory diseases are desirable.
[0004] TNF-.alpha. is a protein whose synthesis occurs in many cell
types in response to an external stimulus, such as, for example, a
mitogen, an infectious organism, or trauma. Signaling from the cell
surface to the nucleus proceeds via several intracellular mediators
including kinases that catalyze phosphorylation of proteins
downstream in the signaling cascade. Important mediators for the
production of TNF-.alpha. cytokine are the mitogen-activated
protein (MAP) kinases, and in particular, p38 kinase.
[0005] p38 Kinases are activated in response to various stress
stimuli, including, but not limited to, proinflammatory cytokines,
endotoxin, ultraviolet light, and osmotic shock. Activation of p38
requires dual phosphorylation by upstream MAP kinase kinases (MKK3
and MKK6) on threonine and tyrosine within a Thr-Gly-Tyr motif,
characteristic of p38 isozymes.
[0006] Four iso-forms of p38 have been described. The .alpha. and
.beta. forms are expressed in inflammatory cells and are thought to
be key mediators of TNF-.alpha. production. Inhibition of the
enzymes p38.alpha. and .beta. in cells results in reduced levels of
expression of TNF-.alpha., and such inhibitors are effective in
animal models of inflammatory disease.
[0007] Molecular cloning of human p38.alpha. identified two
isozymes, which are the splice variant product of a single gene.
Three additional gene products have subsequently been identified,
p38.beta., p38.gamma., and p38.delta.. p38 kinases phosphorylate
and activate the transcription factors, ATF-2, MAX, CHOP, and
C/ERPb, suggesting a role of p38 kinases in gene regulation. In
addition, p38 kinases phosphorylate other protein kinases, such as
MAPK activated protein kinase-2/3 (MAPKAP-K2/3, or MK2/3), and
MAP-kinase-interacting kinase 1/2 (MNK1/2). Recently, activation of
MK2 has been shown to be essential for LPS-induced TNF-.alpha.
expression [Kotlyarov et al., Nature Cell Biol., 1:94-97 (1999)].
Mice lacking MK2 exhibit a 90% reduction in the production of
TNF-.alpha. and are resistant to shock induced by LPS. The
reduction in TNF-.alpha. amounts is due not to decreased production
of the TNF-.alpha. mRNA, but rather to diminished production of the
TNF-.alpha. protein, suggesting that MK2 regulates biosynthesis of
TNF-.alpha. at a post-transcriptional level.
[0008] Ample evidence indicates that the p38 pathway serves an
important role in inflammatory process mediated by IL-1 and
TNF-.alpha..
[0009] Small molecule inhibitors of p38 are expected to have
several advantages over protein inhibitors of TNF-.alpha. or IL-1.
p38 inhibitors not only block the production of TNF-.alpha. and
IL-1, but also directly interfere with many of their secondary
biological effects. In addition, small molecule inhibitors are
unlikely to induce immune reaction in patients, and are believed
active following oral administration.
[0010] The present invention provides novel compounds that are
potent and selective inhibitors of p38.alpha. and .beta., and as
such, are also potent inhibitors of TNF-.alpha. expression in human
cells. Compounds of the present invention are useful in the
treatment of p38- and TNF-.alpha. expression-mediated inflammatory
and other disorders, including, but not limited to, bone
resorption, graft vs. host reaction, atherosclerosis, arthritis,
osteoarthritis, rheumatoid arthritis, gout, psoriasis, topical
inflammatory disease states, adult respiratory distress syndrome,
asthma, chronic pulmonary inflammatory disease, cardiac reperfusion
injury, renal reperfusion injury, thrombus, glomerulonephritis,
Chron's disease, ulcerative colitis, inflammatory bowel disease,
multiple sclerosis, endotoxin shock, osteoporosis, Alzheimer's
disease, congestive heart failure and cachexia.
SUMMARY OF THE INVENTION
[0011] The compounds of the present invention are effective as
inhibitors of inappropriate p38 activity, especially iso forms
.alpha. and .beta., and in turn, of cytokine production, and in
particular, of cellular TNF-alpha (TNF-.alpha.) expression.
Accordingly, compounds of the invention are useful for the
inhibition, prevention and suppression of various pathologies
associated with such activity, such as, for example, inflammation,
asthma, arthritis, atherosclerosis, multiple sclerosis, psoriasis,
autoimmune diseases, Alzeheimers disease and congestive heart
failure, among others.
[0012] In one embodiment, the principles of the present invention
provide a compound, including isomers, enantiomers, diastereomers,
tautomers, pharmaceutically acceptable salts, prodrugs and solvates
thereof, represented by Formula (I): 2
[0013] wherein:
[0014] to one or two of W, Y and X are .dbd.N--;
[0015] one of W, Y and X is selected from .dbd.C--CN, .dbd.C--F,
.dbd.C--NO.sub.2, .dbd.C--Br, .dbd.C--NH.sub.2,
.dbd.C--NHC(O)CH.sub.3 and .dbd.C--Cl;
[0016] the remaining W, Y or X is .dbd.CH--;
[0017] V is --NR.sup.5--;
[0018] Z is halogen or --N(R.sup.1)(R.sup.2)
[0019] R.sup.1 and R.sup.2 are the same or different and are
selected from hydrogen, alkyl, substituted alkyl, aryl, substituted
aryl, cycloalkyl, substituted cycloalkyl, heterocyclyl or
substituted heterocyclyl;
[0020] R.sup.5 is hydrogen or alkyl;
[0021] R.sup.6 is 3
[0022] R.sup.7 is hydrogen, alkyl, substituted alkyl, alkoxy, or
halogen;
[0023] R.sup.8 is hydrogen, alkyl, alkyloxy or cyano;
[0024] R.sup.9 is --C(O)R.sup.10 or unsubstituted or substituted
heterocyclyl;
[0025] R.sup.10 is --N(R.sup.31)(R.sup.32);
[0026] R.sup.31 and R.sup.32 are the same or different and are
selected from hydrogen, alkyl, substituted alkyl, alkoxy, aryl,
substituted aryl, cycloalkyl, substituted cycloalkyl, heterocyclyl
or substituted heterocyclyl;
[0027] R.sup.11 is hydrogen, halogen, O--R.sup.35 or
--N(R.sup.12)(R.sup.13);
[0028] R.sup.12 is hydrogen, alkyl, or substituted alkyl;
[0029] R.sup.13 is --(CH.sub.2).sub.mR.sup.14;
[0030] --N(R.sup.12)(R.sup.13) taken together may form a
heterocyclyl or substituted heterocyclyl;
[0031] m is 0, 1, 2 or 3;
[0032] R.sup.14 is hydrogen, alkyl, substituted alkyl,
--C(O)N(R.sup.31)(R.sup.32), --N(R.sup.33)C(O)R.sup.34, aryl,
substituted aryl, cycloalkyl, substituted cycloalkyl, heterocyclyl,
substituted heterocyclyl, heteroaryl, substituted heteroaryl or
4
[0033] R.sup.15 is hydrogen, alkyl or substituted alkyl;
[0034] R.sup.16 is hydrogen or alkyl; or
[0035] R.sup.33 is hydrogen, alkyl, or substituted alkyl;
[0036] R.sup.34 is alkyl, substituted alkyl, aryl or substituted
aryl;
[0037] R.sup.35 is hydrogen or -(lower alkyl)-R.sup.36
[0038] R.sup.36 is N(R.sup.37)(R.sup.38)
[0039] R.sup.37 is hydrogen, alkyl, or substituted alkyl;
[0040] R.sup.38 is -(substituted alkyl)-R.sup.14; and
[0041] N(R.sup.37)(R.sup.38) taken together may form a heterocyclyl
or substituted heterocyclyl.
[0042] Preferred compounds of this invention are those of Formula
(I) including a pharmaceutically acceptable salt thereof
[0043] wherein:
[0044] one or two of W, Y and X are .dbd.N--;
[0045] one of W, Y and X is selected from .dbd.C--CN, .dbd.C--F,
.dbd.C--NO.sub.2, .dbd.C--Br, .dbd.C--NH.sub.2,
.dbd.C--NHC(O)CH.sub.3 and .dbd.C--Cl;
[0046] the remaining W, Y or X is .dbd.CH--;
[0047] V is --NH--;
[0048] Z is --N(R.sup.1)(R.sup.2);
[0049] R.sup.1 and R.sup.2 are the same or different and are
selected from hydrogen, alkyl or substituted alkyl wherein alkyl is
of 1 to 8 carbons;
[0050] R.sup.6 is 5
[0051] R.sup.7 is hydrogen, alkyl of 1 to 4 carbons, alkoxy of 1 to
4 carbons, or halogen;
[0052] R.sup.8 is hydrogen;
[0053] R.sup.9 is --C(O)R.sup.10 or unsubstituted or substituted
heterocyclyl;
[0054] R.sup.10 is --NH.sub.2 or unsubstituted or substituted
--NH-alkyl, --NH-alkoxy, --NH-heterocyclyl, --NH-phenyl, or
--NH--CH.sub.2-phenyl wherein alkyl and alkoxy are of 1 to 6
carbons;
[0055] R.sup.11 is hydrogen, halogen, O--R.sup.35 or
--N(R.sup.12)(R.sup.13), wherein N(R.sup.12)(R.sup.13) taken
together may form a monocyclic heterocyclyl or substituted
heterocyclyl of 5 to 7 atoms containing 1, 2, or 3 additional
nitrogen atoms or wherein
[0056] R.sup.12 is hydrogen;
[0057] R.sup.13 is alkyl of 1 to 4 carbons or 6
[0058] R.sup.15 and R.sup.16 are independently selected from
hydrogen and methyl;
[0059] R.sup.35 is hydrogen or -(lower alkyl)-R.sup.36;
[0060] R.sup.36 is N(R.sup.37)(R.sup.38)
[0061] R.sup.37 is hydrogen, alkyl, or substituted alkyl;
[0062] R.sup.38 is -(substituted alkyl)-R.sup.14; and
[0063] N(R.sup.37)(R.sup.38) taken together may form a heterocyclyl
or substituted heterocyclyl.
[0064] The principles of the present invention also provide methods
of inhibiting TNF-.alpha. expression in a mammal, wherein the
methods comprise administering to the mammal an effective amount of
a compound represented by Formula (I), or a prodrug or salt
thereof. As used herein, inhibiting TNF-.alpha. expression is
intended to include inhibiting, suppressing and preventing
conditions associated with inappropriate TNF-.alpha. expression,
including, but not limited to, inflammation, asthma, arthritis,
atherosclerosis, multiple sclerosis, psoriasis, autoimmune
diseases, Alzheimer's disease and congestive heart failure.
[0065] The principles of the present invention further provide
methods of treating p38 kinase and TNF-.alpha. mediated disorders
in a mammal, the methods comprising administering to a mammal in
need of such treatment, an effective amount of a compound
represented by Formula (I), or a prodrug or salt thereof. As used
herein, a p38 kinase mediated disorder means a disorder associated
with inappropriate p38 kinase activity; a TNF-.alpha. mediated
disorder means a disorder associated with inappropriate TNF-.alpha.
expression. Such disorders include, but are not limited to,
inflammation, asthma, arthritis, atherosclerosis, multiple
sclerosis, psoriasis, autoimmune diseases, Alzheimer.quadrature.s
disease and congestive heart failure.
[0066] Accordingly, the compounds of the invention, as well as
prodrugs or salts thereof, may be used in the manufacture of a
pharmaceutical composition or medicament for the prophylactic or
therapeutic treatment of disease states in mammals. The compounds
of the present invention may be administered as pharmaceutical
compositions as a monotherapy, or in combination with, for example,
other anti-inflammatory, e.g. a steroid or NSAID (non-steroidal
anti-inflammatory drug) and/or immunosuppressive agents. Such
combination therapies can involve the administration of the various
pharmaceuticals as a single dosage form or as multiple dosage forms
administered simultaneously or sequentially.
[0067] Any suitable route of administration may be employed for
providing a patient with an effective amount of a compound of the
present invention. Suitable routes of administration may include,
for example, oral, rectal, nasal, buccal, parenteral (such as,
intravenous, intrathecal, subcutaneous, intramuscular,
intrasternal, intrahepatic, intralesional, intracranial,
intra-articular, and intra-synovial), transdermal (such as, for
example, patches), and the like. Due to their ease of
administration, oral dosage forms, such as, for example, tablets,
troches, dispersions, suspensions, solutions, capsules, soft
gelatin capsules, and the like, may be preferred. Administration
may also be by controlled or sustained release means and delivery
devices. Methods for the preparation of such dosage forms are well
known in the art.
[0068] Pharmaceutical compositions incorporating compounds of the
present invention may include excipients, a pharmaceutically
acceptable carrier, in addition to other therapeutic ingredients.
Excipients such as starches, sugars, microcrystalline cellulose,
diluents, lubricants, binders, coloring agents, flavoring agents,
granulating agents, disintegrating agents, and the like may be
appropriate depending upon the route of administration. Because of
their ease of administration, tablets and capsules represent the
most advantageous oral dosage unit forms. If desired, tablets may
be coated by standard aqueous or nonaqueous techniques.
[0069] The compounds of the present invention may be used in the
form of pharmaceutically acceptable salts derived from inorganic or
organic bases, and hydrates thereof. Included among such base salts
are ammonium salts, alkali metal salts, such as sodium and
potassium salts, alkaline earth metal salts, such as calcium and
magnesium salts, salts with organic bases, such as
dicyclohexylamine salts, N-methyl-D-glucamine, and salts with amino
acids such as arginine, lysine, and so forth.
DETAILED DESCRIPTION OF THE INVENTION
[0070] [1] Thus, in a first embodiment, the present invention
provides a novel compound of Formula (I) including isomers,
enantiomers, diastereomers, tautomers, pharmaceutically acceptable
salts, prodrugs and solvates thereof, comprising: 7
[0071] wherein:
[0072] one or two of W, Y and X are .dbd.N--;
[0073] one of W, Y and X is selected from .dbd.C--CN, .dbd.C--F,
.dbd.C--NO.sub.2, .dbd.C--Br, .dbd.C--NH.sub.2,
.dbd.C--NHC(O)CH.sub.3 and .dbd.C--Cl;
[0074] the remaining W, Y or X is .dbd.CH--;
[0075] V is --NR.sup.5--;
[0076] Z is halogen or --N(R.sup.1)(R.sup.2);
[0077] R.sup.1 and R.sup.2 are the same or different and are
selected from hydrogen, alkyl, substituted alkyl, aryl, substituted
aryl, cycloalkyl, substituted cycloalkyl, heterocyclyl or
substituted heterocyclyl;
[0078] R.sup.5 is hydrogen or alkyl;
[0079] R.sup.6 is 8
[0080] R.sup.7 is hydrogen, alkyl, substituted alkyl, alkoxy, or
halogen;
[0081] R.sup.8 is hydrogen, alkyl, alkyloxy or cyano;
[0082] R.sup.9 is --C(O)R.sup.10 or unsubstituted or substituted
heterocyclyl;
[0083] R.sup.10 is --N(R.sup.31)(R.sup.32)
[0084] R.sup.31 and R.sup.32 are the same or different and are
selected from hydrogen, alkyl, substituted alkyl, alkoxy, aryl,
substituted aryl, cycloalkyl, substituted cycloalkyl, heterocyclyl
or substituted heterocyclyl;
[0085] R.sup.11 is hydrogen, halogen, O--R.sup.35 or
--N(R.sup.12)(R.sup.13);
[0086] R.sup.12 is hydrogen, alkyl, or substituted alkyl;
[0087] R.sup.13 is --(CH.sub.2).sub.mR.sup.14;
[0088] --N(R.sup.12)(R.sup.13) taken together may form a
heterocyclyl or substituted heterocyclyl;
[0089] m is 0, 1, 2 or 3;
[0090] R.sup.14 is hydrogen, alkyl, substituted alkyl,
--C(O)N(R.sup.31 )(R.sup.32), --N(R.sup.33)C(O)R.sup.34, aryl,
substituted aryl, cycloalkyl, substituted cycloalkyl, heterocyclyl,
substituted heterocyclyl, heteroaryl, substituted heteroaryl or
9
[0091] R.sup.15 is hydrogen, alkyl or substituted alkyl;
[0092] R.sup.16 is hydrogen or alkyl; or
[0093] R.sup.33 is hydrogen, alkyl, or substituted alkyl;
[0094] R.sup.34 is alkyl, substituted alkyl, aryl or substituted
aryl;
[0095] R.sup.35 is hydrogen or -(lower alkyl)-R.sup.36;
[0096] R.sup.36 is N(R.sup.37)(R.sup.38);
[0097] R.sup.37 is hydrogen, alkyl, or substituted alkyl;
[0098] R.sup.38 is -(substituted alkyl)-R.sup.14; and
[0099] N(R.sup.37)(R.sup.38) taken together may form a heterocyclyl
or substituted heterocyclyl.
[0100] [2] In a preferred embodiment, the present invention
provides a compound of Formula (I) including isomers, enantiomers,
diastereomers, tautomers, pharmaceutically acceptable salts,
prodrugs and solvates thereof,
[0101] wherein:
[0102] one or two of W, Y and X are .dbd.N--;
[0103] one of W, Y and X is selected from .dbd.C--CN, .dbd.C--F,
.dbd.C--
[0104] NO.sub.2, .dbd.C--Br, .dbd.C--NH.sub.2,
.dbd.C--NHC(O)CH.sub.3 and .dbd.C--Cl;
[0105] the remaining W, Y or X is .dbd.CH--;
[0106] V is --NH--;
[0107] Z is --N(R.sup.1)(R.sup.2);
[0108] R.sup.1 and R.sup.2 are the same or different and are
selected from hydrogen, alkyl or substituted alkyl wherein alkyl is
of 1 to 8 carbons;
[0109] R.sup.6 is 10
[0110] R.sup.7 is hydrogen, alkyl of 1 to 4 carbons, alkoxy of 1 to
4 carbons, or halogen;
[0111] R.sup.8 is hydrogen;
[0112] R.sup.9 is --C(O)R.sup.10 or unsubstituted or substituted
heterocyclyl;
[0113] R.sup.10 is --NH.sub.2 or unsubstituted or substituted
--NH-alkyl, --NH-alkoxy, --NH-heterocyclyl, --NH-phenyl, or
--NH--CH.sub.2-phenyl wherein alkyl and alkoxy are of 1 to 6
carbons;
[0114] R.sup.11 is hydrogen, halogen, O--R.sup.35 or
--N(R.sup.12)(R.sup.13), wherein N(R.sup.12)(R.sup.13) taken
together may form a monocyclic heterocyclyl or substituted
heterocyclyl of 5 to 7 atoms containing 1, 2, or 3 additional
nitrogen atoms or wherein
[0115] R.sup.12 is hydrogen;
[0116] R.sup.13 is alkyl of 1 to 4 carbons or 11
[0117] R.sup.15 and R.sup.16 are independently selected from
hydrogen and methyl;
[0118] R.sup.35 is hydrogen or -(lower alkyl)-R.sup.36;
[0119] R.sup.36 is N(R.sup.37)(R.sup.38)
[0120] R.sup.37 is hydrogen, alkyl, or substituted alkyl;
[0121] R.sup.38 is -(substituted alkyl)-R.sup.14; and
[0122] N(R.sup.37)(R.sup.38) taken together may form a heterocyclyl
or substituted heterocyclyl.
[0123] [3] In a more preferred embodiment, the present invention
provides a compound of Formula (I) including isomers, enantiomers,
diastereomers, tautomers, pharmaceutically acceptable salts,
prodrugs and solvates thereof,
[0124] wherein:
[0125] one or two of W, Y and X are .dbd.N--;
[0126] one of W, Y and X is selected from .dbd.C--CN, .dbd.C--F,
.dbd.C--NO.sub.2, .dbd.C--Br, .dbd.C--NH.sub.2,
.dbd.C--NHC(O)CH.sub.3 and .dbd.C--Cl;
[0127] the remaining W, Y or X is .dbd.CH--;
[0128] V is --NH--;
[0129] Z is --N(R.sup.1)(R.sup.2)
[0130] R.sup.1 and R.sup.2 are the same or different and are
selected from hydrogen or alkyl of 1 to 8 carbons;
[0131] R.sup.6 is 12
[0132] R.sup.7 is hydrogen, methyl, methoxy, Cl, Br, or F;
[0133] R.sup.8 is hydrogen;
[0134] R.sup.9 is --C(O)R.sup.10 or unsubstituted or substituted
heterocyclyl;
[0135] R.sup.10 is --NH.sub.2, or unsubstituted or substituted
--NH-alkyl, --NH-alkoxy, --NH-phenyl, or --NH--CH.sub.2-phenyl
wherein alkyl and alkoxy are of 1 to 6 carbons; and
[0136] R.sup.11 is hydrogen, halogen, O--R.sup.35 or
--N(R.sup.2)(R.sup.13), wherein N(R.sup.12)(R.sup.13) taken
together form a monocyclic heterocyclyl or substituted heterocyclyl
of 5 to 7 atoms containing 1, 2, or 3 additional nitrogen
atoms.
[0137] [4] In another preferred embodiment, the present invention
provides a compound of Formula (I) including isomers, enantiomers,
diastereomers, tautomers, pharmaceutically acceptable salts,
prodrugs and solvates thereof,
[0138] wherein:
[0139] one of W, Y and X is .dbd.N--;
[0140] one of W, Y and X is selected from .dbd.C--CN, .dbd.C--F,
.dbd.C--NO.sub.2, .dbd.C--Br, .dbd.C--NH.sub.2,
.dbd.C--NHC(O)CH.sub.3 and .dbd.C--Cl;
[0141] the remaining W, Y or X is .dbd.CH--;
[0142] V is --NH--;
[0143] Z is --N(R.sup.1)(R.sup.2);
[0144] R.sup.1 and R.sup.2 are the same or different and are
selected from hydrogen or alkyl of 1 to 8 carbons;
[0145] R.sup.6 is 13
[0146] R.sup.7 is hydrogen, methyl, methoxy, Cl, Br, or F;
[0147] R.sup.8 is hydrogen;
[0148] R.sup.9 is --C(O)R.sup.10 or unsubstituted or substituted
heterocyclyl;
[0149] R.sup.10 is --NH.sub.2, or unsubstituted or substituted
--NH-alkyl, --NH-alkoxy, --NH-phenyl, or --NH--CH.sub.2-phenyl
wherein alkyl and alkoxy are of 1 to 6 carbons;
[0150] R.sup.11 is hydrogen, halogen, --O--R.sup.35 or
--N(R.sup.12)(R.sup.13) wherein N(R.sup.12)(R.sup.13) taken
together form a monocyclic heterocyclyl or substituted heterocyclyl
of 5 to 7 atoms containing 1, 2, or 3 additional nitrogen atoms;
and
[0151] R.sup.15 and R.sup.16 are independently selected from
hydrogen and methyl.
[0152] [5] In another more preferred embodiment, the present
invention provides a compound of Formula (I) including isomers,
enantiomers, diastereomers, tautomers, pharmaceutically acceptable
salts, prodrugs and solvates thereof,
[0153] wherein:
[0154] R.sup.10 is --NH.sub.2, unsubstituted or substituted
--NH--CH.sub.3, --NH--C.sub.2H.sub.5, --NH--OCH.sub.3, or
--NH--OC.sub.2H.sub.5.
[0155] [6] In another more preferred embodiment, the present
invention provides a compound of Formula (I) including isomers,
enantiomers, diastereomers, tautomers, pharmaceutically acceptable
salts, prodrugs and solvates thereof,
[0156] wherein:
[0157] R.sup.9 is unsubstituted or substituted triazole, thiazole,
oxadiazole or imidazole.
[0158] [7] In another more preferred embodiment, the present
invention provides a compound of Formula (I) including isomers,
enantiomers, diastereomers, tautomers, pharmaceutically acceptable
salts, prodrugs and solvates thereof,
[0159] wherein:
[0160] R.sup.11 is hydrogen, halogen, --O-(substituted alkyl),
--NH-(substituted alkyl) or 14
[0161] [8] In yet another preferred embodiment, the present
invention provides a compound of Formula (I) including isomers,
enantiomers, diastereomers, tautomers, pharmaceutically acceptable
salts, prodrugs and solvates thereof,
[0162] wherein:
[0163] R.sup.11 is hydrogen, halogen, --O-(substituted alkyl),
--NH-(substituted alkyl) or 15
[0164] [9] In yet another preferred embodiment, the present
invention provides a compound of Formula (I) including isomers,
enantiomers, diastereomers, tautomers, pharmaceutically acceptable
salts, prodrugs and solvates thereof,
[0165] wherein:
[0166] two of W, Y and X are .dbd.N--;
[0167] the remaining W, Y or X is selected from .dbd.C--CN,
.dbd.C--F, .dbd.C--NO.sub.2, .dbd.C--Br, .dbd.C--NH.sub.2,
.dbd.C--NHC(O)CH.sub.3 and .dbd.C--Cl;
[0168] V is --NH--;
[0169] Z is --N(R.sup.1)(R.sup.2)
[0170] R.sup.1 and R.sup.2 are the same or different and are
selected from hydrogen or alkyl of 1 to 8 carbons;
[0171] R.sup.6 is 16
[0172] R.sup.7 is hydrogen, methyl, methoxy, Cl, Br, or F;
[0173] R.sup.8 is hydrogen;
[0174] R.sup.9 is --C(O)R.sup.10 or unsubstituted or substituted
heterocyclyl;
[0175] R.sup.10 is --NH.sub.2, or unsubstituted or substituted
--NH-alkyl, --NH-alkoxy, --NH-phenyl, or --NH--CH.sub.2-phenyl
wherein alkyl and alkoxy are of 1 to 6 carbons;
[0176] R.sup.11 is hydrogen, halogen, --O--R or
--N(R.sup.12)(R.sup.13), wherein N(R.sup.12)(R.sup.13) taken
together may form a monocyclic heterocyclyl or substituted
heterocyclyl of 5 to 7 atoms containing 1, 2, or 3 additional
nitrogen atoms; and
[0177] R.sup.15 and R.sup.16 are independently selected from
hydrogen and methyl.
[0178] [10] In yet another more preferred embodiment, the present
invention provides a compound of Formula (I) including isomers,
enantiomers, diastereomers, tautomers, pharmaceutically acceptable
salts, prodrugs and solvates thereof,
[0179] wherein:
[0180] R.sup.10 is --NH.sub.2, unsubstituted or substituted
--NH--CH.sub.3, --NH--C.sub.2H.sub.5, --NH--OCH.sub.3, or
--NH--OC.sub.2H.sub.5.
[0181] [11] In yet another more preferred embodiment, the present
invention provides a compound of Formula (I) including isomers,
enantiomers, diastereomers, tautomers, pharmaceutically acceptable
salts, prodrugs and solvates thereof,
[0182] wherein:
[0183] R.sup.9 is unsubstituted or substituted triazole, thiazole,
oxadiazole or imidazole.
[0184] [12] In yet another more preferred embodiment, the present
invention provides a compound of Formula (I) including isomers,
enantiomers, diastereomers, tautomers, pharmaceutically acceptable
salts, prodrugs and solvates thereof,
[0185] wherein:
[0186] R.sup.11 is hydrogen, halogen, --O-(substituted alkyl),
--NH-(substituted alkyl) or 17
[0187] [13] In yet another more preferred embodiment, the present
invention provides a compound of Formula (I) including isomers,
enantiomers, diastereomers, tautomers, pharmaceutically acceptable
salts, prodrugs and solvates thereof,
[0188] wherein:
[0189] R.sup.11 is hydrogen, halogen, --O-(substituted alkyl),
--NH-(substituted alkyl) or 18
[0190] [14] In a second preferred embodiment, the present invention
provides a pharmaceutical composition comprising as an active
ingredient, a compound of the invention or a prodrug or salt
thereof, and a pharmaceutically acceptable carrier.
[0191] [15] In a preferred embodiment, the present invention to
provides a pharmaceutical composition further comprising one or
more additional active ingredients.
[0192] [16] In a more preferred embodiment, the present invention
provides a pharmaceutical composition wherein the additional active
ingredient is an anti-inflammatory compound or an immunosuppressive
agent.
[0193] [17] In a more preferred embodiment, the present invention
provides a pharmaceutical composition wherein the additional active
ingredient is chosen from a steroid and an NSAID.
[0194] [18] In a third embodiment, the present invention provides a
method of inhibiting TNF-.alpha. expression in a mammal comprising
administering to the mammal an effective amount of the
pharmaceutical composition of the invention.
[0195] [19] In a preferred embodiment, the present invention
provides a method of treating TNF-.alpha. mediated disorder
comprising administering to a mammal in need of such treatment, an
effective amount of a pharmaceutical composition of the
invention.
[0196] [20] In a more preferred embodiment, the present invention
provides a method of treating TNF-.alpha. mediated disorder,
wherein the TNF-.alpha. mediated disorder is an inflammatory
disorder.
[0197] [21] In a more preferred embodiment, the present invention
provides a method of treating TNF-.alpha. mediated disorder,
wherein the TNF-.alpha. mediated disorder is chosen from bone
resorption, graft vs. host reaction, atherosclerosis, arthritis,
osteoarthritis, rheumatoid arthritis, gout, psoriasis, topical
inflammatory disease states, adult respiratory distress syndrome,
asthma, chronic pulmonary inflammatory disease, cardiac reperfusion
injury, renal reperfusion injury, thrombus, glomerulonephritis,
Chron's disease, ulcerative colitis, inflammatory bowel disease,
multiple sclerosis, endotoxin shock, osteoporosis, Alzheimer's
disease, congestive heart failure and cachexia.
[0198] [22] In a more preferred embodiment, the present invention
provides a method of treating TNF-.alpha. mediated disorder wherein
the pharmaceutical composition of the invention is administered
with one or more additional anti-inflammatory or immunosuppressive
agents as a single dose form or as separate dosage forms.
[0199] [23] In an even more preferred embodiment, the present
invention provides a method of treating a condition associated with
TNF-.alpha. expression in a mammal comprising administering to a
mammal in need of such treatment, an effective amount of a
pharmaceutical composition of the invention.
[0200] [24] In an even more preferred embodiment, the present
invention provides a method of treating a condition associated with
TNF-.alpha. expression in a mammal wherein the condition associated
with TNF-.alpha. expression is an inflammatory disorder.
[0201] [25] In a more preferred embodiment, the present invention
provides a method of treating a condition associated with
TNF-.alpha. expression, wherein the condition associated with
TNF-.alpha. expression is chosen from bone resorption, graft vs.
host reaction, atherosclerosis, arthritis, osteoarthritis,
rheumatoid arthritis, gout, psoriasis, topical inflammatory disease
states, adult respiratory distress syndrome, asthma, chronic
pulmonary inflammatory disease, cardiac reperfusion injury, renal
reperfusion injury, thrombus, glomerulonephritis, Crohn's disease,
ulcerative colitis, inflammatory bowel disease, multiple sclerosis,
endotoxin shock, osteoporosis, Alzheimer's disease, congestive
heart failure and cachexia.
[0202] [26] In a more preferred embodiment, the present invention
provides a method of treating a condition associated with
TNF-.alpha. expression wherein the pharmaceutical composition of
the invention is administered with one or more additional
anti-inflammatory or immunosuppressive agents as a single dose form
or as separate dosage forms.
[0203] [27] In another more preferred embodiment, the present
invention provides a method of treating a condition associated with
p38 kinase activity in a mammal comprising administering to a
mammal in need of such treatment, an effective amount of a
pharmaceutical composition of the invention.
[0204] [28] In another more preferred embodiment, the present
invention provides a method of treating a condition associated with
p38 kinase activity in a mammal wherein the condition associated
with p38 activity is an inflammatory disorder.
[0205] [29] In a more preferred embodiment, the present invention
provides a method of treating a condition associated with p38
kinase activity is chosen from bone resorption, graft vs. host
reaction, atherosclerosis, arthritis, osteoarthritis, rheumatoid
arthritis, gout, psoriasis, topical inflammatory disease states,
adult respiratory distress syndrome, asthma, chronic pulmonary
inflammatory disease, cardiac reperfusion injury, renal reperfusion
injury, thrombus, glomerulonephritis, Chron's disease, ulcerative
colitis, inflammatory bowel disease, multiple sclerosis, endotoxin
shock, osteoporosis, Alzheimer's disease, congestive heart failure
and cachexia.
[0206] [30] In a more preferred embodiment, the present invention
provides a method of treating a condition associated with p38
activity wherein the pharmaceutical composition of the invention is
administered with one or more additional anti-inflammatory or
immunosuppressive agents as a single dose form or as separate
dosage forms.
[0207] [31] In a fourth embodiment, the present invention provides
a compound including isomers, enantiomers, diastereomers,
tautomers, pharmaceutically acceptable salts, prodrugs and solvates
selected from: 1920212223242526272829303132333435
Abbreviations & Definitions
[0208] The following terms and abbreviations retain the indicated
meaning throughout this disclosure.
[0209] ATP=adenosine triphosphate
[0210] cDNA=complementary DNA
[0211] DCE=dichloroethylene
[0212] DCM=dichloromethane=methylene chloride=CH.sub.2Cl.sub.2
[0213] DIC=diisopropylcarbodiimide
[0214] DIEA=N,N-diisopropylethylamine
[0215] DMF=N,N-dimethylformamide
[0216] DMSO=dimethyl sulfoxide
[0217] DTT=dithiothreitol
[0218] EDTA=ethylenediaminetetraacetic acid
[0219] EIA=enzyme immunoassay
[0220] ELISA=enzyme-linked immunosorbent assay
[0221] Fmoc=9-fluorenylmethoxycarbonyl
[0222] GST=glutathione S-transferase
[0223] HOBt=1-hydroxybenzotriazole
[0224] LPS=lipopolysaccharide
[0225] MBP=myelin basic protein
[0226] MES=2-(N-morpholino)ethanesulfonic acid
[0227] mRNA=messenger RNA
[0228] PCR=polymerase chain reaction
[0229] Pr.sub.2NEt=dipropylethylamine
[0230] i-Pr.sub.2NEt=diisopropylethylamine
[0231] RPMI=Roswell Park Memorial Institute
[0232] TBS=t-butyldimethylsilyl
[0233] TFA=trifluoroacetic acid
[0234] THF=tetrahydrofuran
[0235] "Alkyl" is intended to include linear or branched
hydrocarbon structures and combinations thereof of 1 to 20 carbons.
"Lower alkyl" means alkyl groups of from 1 to about 10, preferably
from 1 to about 8, and more preferably, from 1 to about 6 carbon
atoms. Examples of such radicals include methyl, ethyl, n-propyl,
isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, pentyl, iso-amyl,
hexyl, octyl and the like.
[0236] "Aryl" means an aromatic hydrocarbon radical of 6 to about
16 carbon atoms, preferably of 6 to about 12 carbon atoms, and more
preferably of 6 to about 10 carbon atoms. Examples of aryl groups
are phenyl, which is preferred, 1-naphthyl and 2-naphthyl.
[0237] "Cycloalkyl" refers to saturated hydrocarbon ring structures
of from 3 to 12 carbon atoms, and preferably from 3 to 6 carbon
atoms. Examples include cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, norbornyl, adamantyl, and the like. "Lower cycloalkyl",
refers to cycloalkyl of 3 to 6 carbons.
[0238] "Heterocyclyl" refers to saturated, partially saturated or
unsaturated monocyclic structures of from 3 to 8 atoms, preferably
5 or 6 atoms, and bicyclic structures of 9 or 10 atoms containing
one or more carbon atoms and from 1 to 4 heteroatoms chosen from O,
N, and S. The point of attachment of the heterocyclyl structure is
at an available carbon or nitrogen atom. Examples include:
imidazole, pyridine, indole, thiophene, benzopyranone, thiazole,
furan, benzimidazole, quinoline, isoquinoline, quinoxaline,
pyrimidine, pyrazine, tetrazole, pyrazole, pyrrolyl, pyridinyl,
pyrazolyl, triazolyl, pyrimidinyl, pyridazinyl, oxazolyl,
thiazolyl, imidazolyl, indolyl, thiophenyl, furanyl, tetrazolyl,
2-pyrrolinyl, 3-pyrrolinyl, pyrrolindinyl, 1,3-dioxolanyl,
imidazolinyl, imidazolidinyl, pyrazolinyl, pyrazolidinyl,
isoxazolyl, isothiazolyl, 1,2,3-oxadiazolyl, 1,2,3-triazolyl,
1,2,4-triazolyl, 1,3,4-thiadiazolyl, 2H-pyranyl, 4H-pyranyl,
piperidinyl, 1,4-dithianyl, thiomorpholinyl, pyrazinyl,
piperazinyl, 1,3,5-triazinyl, 1,2,5-trithianyl, benzo(b)thiophenyl,
benzimidazolyl, quinolinyl, and the like.
[0239] "Alkoxy" means a straight, branched or cyclic hydrocarbon
configuration and combinations thereof, including from 1 to 20
carbon atoms, preferably from 1 to 8 carbon atoms, more preferably
from 1 to about 4 carbon atoms, and an oxygen atom at the point of
attachment. Suitable alkoxy groups include methoxy, ethoxy,
n-propoxy, isopropoxy, n-butoxy, iso-butoxy, s-butoxy, t-butoxy,
cyclopropyloxy, cyclohexyloxy, and the like. "Lower alkoxy" refers
to alkoxy groups having from 1 to 4 carbon atoms. Similarly,
"alkylthio" refers to such groups having a sulfur atom at the point
of attachment.
[0240] "Alkenyl" refers to an unsaturated acyclic hydrocarbon
radical in so much as it contains at least one double bond. "Lower
alkenyl" refers to such radicals containing from about 2 to about
10 carbon atoms, preferably from about 2 to about 8 carbon atoms
and more preferably 2 to about 6 carbon atoms. Examples of suitable
alkenyl radicals include propenyl, buten-1-yl, isobutenyl,
penten-1-yl, 2-methylbuten-1-yl, 3-methylbuten-1-yl, hexen-1-yl,
hepten-1-yl, and octen-1-yl, and the like.
[0241] "Alkynyl" refers to an unsaturated acyclic hydrocarbon
radical containing at least one triple bond. Examples include
ethynyl, propynyl, and the like.
[0242] "Substituted alkyl" means an alkyl wherein one or more
hydrogens, preferably one, two, or three hydrogens, attached to an
aliphotic carbon are replaced with a substituent such as
--N(R.sup.31)(R.sup.32), alkoxy, alkylthio, halogen, cyano,
carboxyl, hydroxyl, --SO.sub.2-alkyl, --CO.sub.2-alkyl,
--C(O)-alkyl, nitro, cycloalkyl, substituted cycloalkyl, aryl,
substituted aryl, heterocyclyl, substituted heterocyclyl,
--C(O)--N(R.sup.31)(R.sup.32), or --NH--C(O)-alkyl. Examples of
such substituent groups include methoxy, ethoxy, propoxy, amino,
methylamino, dimethylamino, phenyl naphthyl, chlorine, fluorine,
and the like.
[0243] "Substituted cycloalkyl" means a cycloalkyl wherein one or
more hydrogens, preferably one, two or three hydrogens, attached to
a ring carbon are replaced with a substituent such as alkyl,
substituted alkyl, --N(R.sup.31)(R.sup.32), alkoxy, alkylthio,
aryl, substituted aryl, halogen, cyano, carboxyl, hydroxyl, nitro,
--SO.sub.2-alkyl, --CO.sub.2-alkyl, --C(O)-alkyl,
--C(O)--N(R.sup.3)R.sup.32), or --NH--C(O)-alkyl. Examples of such
groups include methyl, isopropyl, methoxy, ethoxy, propoxy, amino,
methylamino, dimethylamino, phenyl, chlorine, fluorine and the
like. Also included within this definition are cycloalkyl rings
having a fused aryl, preferably phenyl, or cycloalkyl such as
36
[0244] and the like.
[0245] "Substituted aryl" means an aryl wherein one or more
hydrogens, preferably one, two or three hydrogens, attached to an
aromatic carbon are replaced with a substituent such as alkyl,
substituted alkyl, --N(R.sup.31)(R.sup.32) alkoxy, alkylthio, aryl,
substituted aryl, halogen, cyano, nitro, carboxyl, hydroxyl,
--SO.sub.2-alkyl, --CO.sub.2-alkyl, --C(O)-alkyl,
--C(O)--N(R.sup.3)(R.sup.32), or --NH--C(O)-alkyl. Examples of such
substituents include methyl, isopropyl, methoxy, ethoxy, propoxy,
amino, methylamino, dimethylamino, phenyl, chlorine, fluorine,
--CO.sub.2CH.sub.3, --C(O)--NH.sub.2, and the like.
[0246] "Substituted heterocyclyl" means a heterocyclyl substituted
at one or more available carbon or nitrogen atoms, preferably at
one or two carbon and/or nitrogen atoms, with a substituent such as
alkyl, substituted alkyl, --N(R.sup.31)(R.sup.32), alkoxy,
alkylthio, aryl, substituted aryl, halogen, cyano, nitro, oxo,
carboxyl, hydroxyl, --SO.sub.2-alkyl, --CO.sub.2-alkyl,
--C(O)-alkyl, --C(O)--N(R.sup.31)(R.s- up.32), or --NH--C(O)-alkyl.
Examples of such groups include methyl isopropyl, methoxy, ethoxy,
propoxy, amino, methylamino, dimethylamino, phenyl, chlorine,
fluorine and the like.
[0247] "Halogen" is intended to include for example, F, Cl, Br and
I.
[0248] The term "prodrug" refers to a chemical compound that is
converted to an active agent by metabolic processes in vivo. [See,
e.g., N. Boder and J. J. Kaminski, Ann. Rep. Med. Chem. 22:303
(1987) and H. Bundgarrd, Adv. Drug Delivery Rev., 3:39 (1989)].
With regard to the present invention, a prodrug of a compound of
Formula I is intended to mean any compound that is converted to a
compound of Formula I by metabolic processes in vivo. The use of
prodrugs of compounds of Formula I in any of the methods described
herein is contemplated and is intended to be within the scope of
the invention.
[0249] Terminology related to "protected," "protecting" and/or
"deprotecting" functionalities is used throughout this application.
Such terminology is well understood by persons of skill in the art
and is used in the context of processes which involve sequential
treatment with a series of reagents. In this context, a protecting
group refers to a group which is used to mask a functionality
during a process step in which it would otherwise react, but in
which reaction is undesirable. The protecting group prevents
reaction at that step, but may be subsequently removed to expose
the original functionality. The removal or "deprotection" occurs
after the completion of the reaction or reactions in which the
functionality would interfere. Thus, when a sequence of reagents is
specified, as it is in the processes of the invention, the person
of ordinary skill can readily envision those groups that would be
suitable as "protecting groups" for the functionalities
involved.
[0250] In the case of the present invention, the typical
functionalities that must be protected are amines. Suitable groups
for that purpose are discussed in standard textbooks in the field
of chemistry, such as Protective Groups in Organic Synthesis by T.
W. Greene [John Wiley & Sons, New York, 1991], which is
incorporated herein by reference. Particular attention is drawn to
the chapter entitled "Protection for the Amino Group" (pages
309-405). Preferred protecting groups include BOC and Fmoc.
Exemplary methods for protecting and deprotecting with these groups
are found in Greene and Wuts on pages 318 and 327.
Optical Isomers--Diastereomers--Geometric Isomers
[0251] Some of the compounds described herein contain one or more
asymmetric centers and may thus give rise to enantiomers,
diastereomers, and other stereoisometric forms which may be defined
in terms of absolute stereochemistry as (R)-- or (S)--, or as (D)--
or (L)-- for amino acids. The present invention is meant to include
all such possible diastereomers as well as 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 optically resolved using conventional techniques. When
the compounds described herein contain olefinic double bonds or
other centers of geometric asymmetry, and unless specified
otherwise, it is intended to include both (E)-- and (Z)-- geometric
isomers. Likewise, all tautomeric forms are intended to be
included.
[0252] Compounds of the invention which incorporate chiral diamines
may be resolved into pairs of enantiomers by known techniques.
Where pure enantiomers of starting materials are not commercially
available, they may be obtained by classic resolution, which may
employ, for example, fractional crystallization of diastereomeric
salts. Compounds of the invention may have more than one chiral
center, for example wherein reductive amination of a homochiral
intermediate leads to a mixture of diastereomers. Racemic
intermediates and compounds of the invention may also be resolved
by chromatographic separation, such as for example, HPLC using a
column loaded with a homochiral support, to yield pure isomeric
compounds.
[0253] The configuration of any carbon-carbon double bond appearing
herein is selected for convenience only and is not intended to
designate a particular configuration; thus a carbon-carbon double
bond depicted arbitrarily herein as trans may be cis, trans, or a
mixture of the two in any proportion.
[0254] In view of the above definitions, other chemical terms used
throughout this application can be easily understood by those of
skill in the art. Terms may be used alone or in any combination
thereof. The preferred and more preferred chain lengths of the
radicals apply to all such combinations.
Utility
[0255] The compounds of the present invention have demonstrated
utility as selective inhibitors of inappropriate p38 kinase
activity, and in particular, isoforms p38.alpha. and p38.beta.. As
such, compounds of the present invention have utility in the
treatment of conditions associated with inappropriate p38 kinase
activity. Such conditions include diseases in which cytokine levels
are modulated as a consequence of intracellular signaling via p38,
and in particular, diseases that are associated with an
overproduction of such cytokines as Il-1, Il-4, IL-8, and in
particular, TNF-.alpha..
[0256] As inhibitors of p-38 kinase activity, compounds of the
present invention are useful in the treatment and prevention of
p-38 mediated conditions including, but not limited to,
inflammatory diseases, autoimmune diseases, destructive bone
disorders, proliferative disorders, angiogenic disorders,
infectious diseases, neurodegenerative diseases, viral diseases,
allergies, myocardial ischemia, reperfusion/ischemia in stroke,
heart attacks, organ hypoxia, vascular hyperplasia, cardiac
hypertrophy, thrombin-induced platelet aggregation, and conditions
associated with prostaglandin endoperoxidase synthase-2.
[0257] Inflammatory diseases which may be treated or prevented
include, but are not limited to, acute pancreatitis, chronic
pancreatitis, asthma, allergies and adult respiratory distress
syndrome.
[0258] Autoimmune diseases which may be treated or prevented
include, but are not limited to, glomerulonephritis, rheumatoid
arthritis, systemic lupus erythematosis, scleroderma, chronic
thyroiditis, Grave.quadrature.s disease, autoimmune gastritis,
diabetes, autoimmune hemolytic anemia, autoimmune neutropenia,
thrombocytopenia, atopic dermatitis, chronic active hepatitis,
myasthenia gravis, multiple sclerosis, inflammatory bowel disease,
ulcerative colitis, Crohn's disease, psoriasis, or graft vs. host
disease.
[0259] Destructive bone disorders which may be treated or prevented
include, but are not limited to, osteoporosis, osteoarthritis and
multiple myeloma-related bone disorder.
[0260] Proliferative diseases which may be treated or prevented
include, but are not limited to, acute myelogenous leukemia,
chronic myelogenous leukemia, metastatic melanoma, Kaposi's
sarcoma, and multiple myeloma.
[0261] Infectious diseases which may be treated or prevented
include, but are not limited to, sepsis, septic shock, and
Shigellosis.
[0262] Neurodegenerative diseases which may be treated or prevented
by the compounds of this invention include, but are not limited to,
Alzheimer's disease, Parkinson's disease, cerebral ischemias or
neurodegenerative disease caused by traumatic injury.
[0263] Angiogenic disorders which may be treated or prevented
include solid tumors, ocular neovasculization, infantile
haemangiomas.
[0264] Viral diseases which may be treated or prevented include,
but are not limited to, acute hepatitis infection (including
hepatitis A, hepatitis B and hepatitis C), HIV infection and CMV
retinitis.
[0265] In addition, p38 inhibitors of this invention also exhibit
inhibition of the expression of inducible pro-inflammatory proteins
such as prostaglandin endoperoxide synthase-2 (PGHS-2), also
referred to as cyclooxygenase-2 (COX-2). Accordingly, additional
p38 mediated conditions include edema, analgesia, fever and pain,
such as neuromuscular pain, headache, pain caused by cancer, dental
pain and arthritis pain.
[0266] As a result of their p38 inhibitory activity, compounds of
the present invention have utility in the treatment and prevention
of diseases associated with cytokine production. For example,
compounds of the present invention are useful in the treatment and
prevention of:
[0267] Il-1 mediated diseases such as, for example, rheumatoid
arthritis, osteoarthritis, stroke, endotoxemia and/or toxic shock
syndrome, inflammatory reaction induced by endotoxin, inflammatory
bowel disease, tuberculosis, atherosclerosis, muscle degeneration,
cachexia, psoriatic arthritis, Reiter's syndrome, gout, traumatic
arthritis, rubella arthritis, acute synovitis, diabetes, pancreatic
.beta.-cell disease and Alzheimer's disease;
[0268] IL-8 mediated diseases or conditions such as, for example,
those characterized by massive neutrophil infiltration, such as
psoriasis, inflammatory bowel disease, asthma, cardiac and renal
reperfusion injury, adult respiratory distress syndrome, thrombosis
and glomerulonephritis; and
[0269] TNF-mediated diseases or conditions such as rheumatoid
arthritis, rheumatoid spondylitis, osteoarthritis, gouty arthritis
and other arthritic conditions, sepsis, septic shock syndrome,
adult respiratory distress syndrome, cerebral malaria, chronic
pulmonary inflammatory disease, silicosis, pulmonary sarcoisosis,
bone resorption disease, reperfusion injury, graft vs. host
reaction, allograft rejections, fever and myalgias due to
infection, cachexia secondary to infection, AIDS, ARC or
malignancy, meloid formation, scar tissue formation, Crohn's
disease, ulcerative colitis, pyresis, viral infections, such as
HIV, CMV, influenza and herpes; and veterinary viral infections,
such as lentivirus infections, including, but not limited to equine
infectious anemia virus; or retro virus infections, including
feline immunodeficiency virus, bovine immunodeficiency virus, or
canine immunodeficiency virus.
[0270] The compounds of formula I including a pharmaceutically
acceptable salt or hydrate thereof may be administered by any
suitable route as described previously to treat the above mentioned
diseases and conditions. The method of administration will, of
course, vary depending upon the type of disease being treated. The
amount of active compound administered will also vary according to
the method of administration and the disease being treated. An
effective amount will be within the dosage range of about 0.1 to
about 100 mg/kg, preferably about 0.2 to about 50 mg/kg, in a
single or multiple doses administered at appropriate intervals
throughout the day.
[0271] The IC.sub.50 values (concentration required to inhibit 50%
of specific binding) of compounds of the present invention for
inhibition of p38 activity are below 5 .mu.M. Preferred compounds
have an IC.sub.50 below 1 .mu.M.
Biological Assays
Generation of p38 Kinases
[0272] cDNAs of human p38.alpha., .beta. and .gamma. isozymes were
cloned by PCR. These cDNAs were subcloned in the pGEX expression
vector (Pharmacia). GST-p38 fusion protein was expressed in E. Coli
and purified from bacterial pellets by affinity chromatography
using glutathione agarose. p38 fusion protein was activated by
incubating with constitutively active MKK6. Active p38 was
separated from MKK6 by affinity chromatography. Constitutively
active MKK6 was generated according to Raingeaud et al. [Mol. Cell.
Biol., 1247-1255 (1996)].
[0273] TNF-.alpha. Production by LPS-stimulated PBMCS
[0274] Heparinized human whole blood was obtained from healthy
volunteers. Peripheral blood mononuclear cells (PBMCs) were
purified from human whole blood by Ficoll-Hypaque density gradient
centrifugation and resuspended at a concentration of
5.times.10.sup.6/ml in assay medium (RPMI medium containing 10%
fetal bovine serum). 50 .mu.l of cell suspension was incubated with
50 .mu.l of test compound (4.times. concentration in assay medium
containing 0.2% DMSO) in 96 well-tissue culture plates for 5
minutes at room temperature. 100 .mu.l of LPS (200 ng/ml stock) was
then added to the cell suspension and the plate was incubated for 6
hours at 37.degree. C. Following incubation, the culture medium was
collected and stored at -20.degree. C. TNF.alpha. concentration in
the medium was quantified using a standard ELISA kit
(Pharmingen-San Diego, Calif.). Concentrations of TNF.alpha. and
IC50 values for test compounds (concentration of compound that
inhibited LPS-stimulated TNF.alpha. production by 50%) were
calculated by linear regression analysis.
LPS-induced TNF Production in THP-1 Cells
[0275] Human monocytic THP-1 cells were maintained in RPMI 1640
medium supplemented with 10% fetal bovine serum. Cells (40,000
cells in 80 .mu.l) were added to wells of 96-well flat-bottomed
plates. Tested compounds (10 .mu.l) or vehicle (3% DMSO) were added
to wells. Subsequently, LPS (Sigma, #L7261; 10 .mu.l/well) was
added to the cells for a final concentration of 1 .mu.l/mL. Plates
were incubated overnight at 37.degree. C. and 5% CO.sub.2.
Supernatant (50 .mu.l/well) was harvested for an ELISA assay. TNF
was captured by an anti-human TNF antibody (R&D, #MAB610) which
was pre-absorbed in high binding EIA plates (Costar, #3590).
Captured TNF was recognized by a biotinlated anti-human TNF
polyclonal antibody (R&D, #BAF210). Streptavidin conjugated
with peroxidase was added to each well, and the activity of
peroxidase was quantitated by a peroxide substrate kit (Pierce,
#34062 and #34006).
p38 Assay
[0276] The assays were performed in V-bottomed 96-well plates. The
final assay volume was 60 .mu.l prepared from three 20 .mu.l
additions of enzyme, substrates (MBP and ATP) and test compounds in
assay buffer (50 mM Tris pH 7.5, 10 mM MgCl.sub.2, 50 mM NaCl and 1
mM DTT). Bacterially expressed, activated p38 was pre-incubated
with test compounds for 10 min. Prior to initiation of reaction
with substrates. The reaction was incubated at 25.degree. C. for 45
min. and terminated by adding 5 .mu.l of 0.5 M EDTA to each sample.
The reaction mixture was aspirated onto a pre-wet filtermat using a
Skatron Micro96 Cell Harvester (Skatron, Inc.), then wash with PBS.
The filtermat was then dried in a microwave oven for 1 min.,
treated with MeltilLex A scintillation wax (Wallac), and counted on
a Microbeta scintillation counter Model 1450 (Wallac). Inhibition
data were analyzed by nonlinear least-squares regression using
prizm (Graphpad Software). The final concentration of reagents in
the assays are ATP, 1 .mu.M; [.gamma.-.sup.33P]ATP, 3 nM,; MBP
(Sigma, # M1891), 2 .mu.g/well; p38, 10 nM; and DMSO, 0.3%.
Methods of Synthesis
[0277] General methods of synthesis for compounds of the present
invention are illustrated by the following examples. Compounds of
the invention may be prepared by standard techniques known in the
art, involving both solution and solid phase chemistry. Starting
materials are commercially available or may by readily prepared by
one of skill in the art with known methods, or by methods disclosed
herein. Specific embodiments described are presented by way of
illustration only, and the invention is not limited thereto.
Modifications and variations in any give material or process step
will be readily apparent to one of skill in the art and all are to
be included within the scope of the invention.
[0278] As illustrated in Scheme 1 and Scheme 2, compounds of
Formula I wherein V is --NR.sup.5--; one or two of W, X and Y are
N; and each of Z and R.sup.11 are attached to the core pyrimidine
or pyridine by --N-- or --O--, may be prepared from
trihalopyrimidine by sequential reactions with three different
amines (1, 2, 3), or two different amines (1, 2) and an alcohol,
and subsequent introduction of an additional substituent on the
pyrimidine core. An alternative method of preparation may start
from dihalocyano-methylsulfanyl-pyrimidine (Scheme 2). Preferably,
one of the amines will be an aniline and another will be a diamine
suitably protected on its distal N. The person of skill will
recognize that the amines themselves, the sequence of the three
substitutions, as well as the position of the nitrile may be
varied, and are not limited by the particular example shown in
Scheme 1 or Scheme 2. 37 38
[0279] With respect to Formula I of the invention, Amine 1
corresponds to --N(R.sup.5)(R.sup.6); Amine 2 corresponds to --Z;
and Amine 3 corresponds to --R.sup.11 and such designations are
used interchangeably in the description below.
[0280] Preparation of Amines
3-(4-Methyl-3-nitro-phenyl)-4H-[1,2,4]triazole
[0281] 39
[0282] Hydrogen chloride was bubbled through a solution of
3-nitro-p-tolunitrile (0.49 g, 3 mmol) in 40 mL of ethanol at room
temp for 10 min. The solution was continued stirring at room temp
for 60 min and the solvent was then evaporated under vacuum to
dryness to give a white solid. 40
[0283] The intermediate so obtained was dissolved in 20 mL of
ethanol, neutralized with sodium ethoxide solution and the
resulting precipitate was removed by filtration. To the filtrate
was added at room temp formic hydrazide (0.2 g, 3 mmol) and the
solution was continued stirring at room temp for 2 h. After removal
of volatiles in vacuo, the residue was dissolved in 30 mL of
m-xylene and refluxed at 150.degree. C. for 16 h. Removal of
volatiles in vacuo and purification using flash chromatography
afforded 0.26 g of the final product. (Yield: 43%). MS (m/z) calcd
for C.sub.9H.sub.8N.sub.4O.sub.2 (MH+) 205.2, found, 205.1.
[0284] Coupling of Substituted Pyridines with Amines
2-Chloro-6-[(2,2-dimethyl-propyl)-methyl-amino]-5-fluoro-nicotinonitrile
[0285] 41
[0286] A solution of 2,6-dichloro-3-cyano-5-fluoropyridine (1.0 g,
5.23 mmol), N-methyl -neopentylamine hydrochloride (830 mg, 6.0
mmol) and triethylamine (1.6 mL) in acetonitrile (20 mL) was
stirred at room temp for 4 hours. Then volatiles were removed in
vacuo and the residue was partitioned between ethyl acetate and
water. The organic layer was dried (sodium sulfate) and
concentrated in vacuo to afford the product (1.11 g, 83%).
C.sub.12H.sub.15ClFN.sub.3 MS m/e=256 (M+H)
[0287] Preparation of Substituted Pyrimidines
4,6-Dichloro-2-methylsulfanyl-pyrimidine-5-carbaldehyde
[0288] 42
[0289] To phosphoryl chloride (108 mL) chilled in ice bath was
added dimethylformamide (35 mL). The mixture was allowed to stand
at 20 degree for one hour, then 25 g of
2-methylsulfanyl-pyrimidine-4,6-diol was added slowly. After 30
minutes, the reaction mixture was heated to 100.degree. C. for 6
hours. The reaction mixture was poured onto crushed ice and the
precipitate was collected by filtration. The crude product was
purified with flash chromatography to afford 11.13 g of
4,6-Dichloro-2-methylsulfa- nyl-pyrimidine-5-carbaldehyde (Yield
=32%). .sup.1H-NMR (300 MHz, CDCl.sub.3): .delta.10.50 (s, 1H),
2.76(s, 3H).
4,6-Dichloro-2-methylsulfanyl-pyrimidine-5-carbaldehyde oxime
[0290] 43
[0291] 4,6-Dichloro-2-methylsulfanyl-pyrimidine-5-carbaldehyde
(7.34 g, 33.09 mmol), hydroxylamine hydrochloride (2.31 g, 33.33
mmol), acetic acid (49.6 mL), and water (3.3 mL) were mixed, and
heated to 60.degree. C. for 2 hours. The reaction mixture was
diluted with water and cooled under ice bath. The precipitate was
collected and dried (Yield =6.41 g, 82%). MS (m/z): 238 (M+H).
4,6-Dichloro-2-methylsulfanyl-pyrimidine-5-carbonitrile
[0292] 44
[0293] 4,6-Dichloro-2-methylsulfanyl-pyrimidine-5-carbaldehyde
oxime (7.20 g, 30.38 mmol) was added to neat thionyl chloride
(29.63 g, 245 mmol), then the mixture was heated to reflux for 4
hours. The reaction mixture was poured onto ice-water. The
precipitate of 4,6-dichloro-2-methylsulfan-
yl-pyrimidine-5-carbonitrile was collected and dried (Yield 6.15 g,
92%). .sup.1H-NMR (300 MHz, CDCl.sub.3): .delta.2.75(s, 3H) 45
3-(6-Chloro-5-cyano-2-methylsulfanyl-pyrimidin-4-ylamino)-4,N-dimethyl-ben-
zamide
[0294] 4,6-Dichloro-2-methylsulfanyl-pyrimidine-5-carbonitrile
(2.19 g, 10 mmol), 3-amino-4,N-dimethyl-benzamide(1.64 g, 10 mmol),
and DIEA(1.40 g, 18.8 mmol) were mixed in THF (20 mL). The
resulting mixture was stirred at room temperature for overnight.
The solvent was evaporated and the residue was partitioned between
ethyl acetate and water. The organic layer was concentrated and the
crude product was purified by flash chromatography to obtain
3-(6-chloro-5-cyano-2-methylsulfanyl-pyrimidin-4-
-ylamino)-4,N-dimethyl-benzamide (2.78 g, 80%). MS (m/z): 348
(M+H).
[0295] The following examples illustrate preferred embodiments of
the present invention and do not limit the scope of the present
invention, which is defined in the claims.
[0296] Compounds shown in Tables 1 and 2 have been synthesized
according to the methods described herein and have been tested in
accordance with the protocols described below. These compounds are
provided by way of illustration only, and the invention is not
intended to be limited thereto. Exemplary syntheses of some
compounds are also provided.
1TABLE 1 Ex # m/z R.sub.t 1 46 367 4.67 2 47 420, 422 3 48 433 3.09
4 49 405 2.58 5 50 499 6 51 466 4.47 7 52 452 4.22 8 53 452 4.16 9
54 424 3.56 10 55 438 3.94 11 56 435 12 57 486 13 58 432 14 59 466
15 60 449 16 61 466 17 62 376 18 63 475 7.3 19 64 405 6.2 20 65 335
4.6 21 66 445 22 67 496 4.64 23 68 383 10.3 24 69 357 25 70 399 26
71 396 27 72 500 11.0 28 73 366 29 74 510 4.81 30 75 509 4.84 31 76
509 4.28 32 77 412 6.0 33 78 467 34 79 35 80 405 5.96 36 81 480
4.67 37 82 496 5.06 38 83 494 4.83 39 84 468 4.69 40 85 396 41 86
479 42 87 495 43 88 460 5.51 44 89 494 4.84 45 90 495 4.46 46 91
493 4.20 47 92 422 48 93 399 7.5 49 94 406, 409 5.7 50 95 353 6.0
51 96 367 6.4 52 97 413 7.7 53 98 367 4.53 54 99 389 55 100 403 56
101 387 6.9 57 102 373 7.0 58 103 451 59 104 451 60 105 451 61 106
509 12.5 62 107 495 12.4
[0297]
2TABLE 2 Example #70 108 Example #71 109 Example #72 110 Example
#73 111 Example #74 112 Example #75 113 Example #76 114 Example #77
115 Example #78 116
EXAMPLE 70
[0298] 117
Synthesis of
3-{3-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-5-fluoro-py-
ridin-2-ylamino }-N-methoxy-4-methyl-benzamide
[0299] A mixture of 2-chloro-6-[(2,2-dimethyl
-propyl)-methyl-amino]-5-flu- oro-nicotinonitrile (120 mg, 0.47
mmol), 3-amino-N-methoxy-4-methyl-benzam- ide (120 mg, 0.66 mmol)
and potassium fluoride (30 mg, 0.51 mmol) in DMSO (1 mL) was heated
to 150.degree. C. overnight. The reaction mixture was allowed to
cool down to room temp and then partitioned between water and ethyl
acetate. The organic layer was dried (sodium sulfate) and
concentrated under reduced pressure. The product (7.5 mg, 4%) was
obtained after purification by silica gel chromatography with 30%
EtOAc in hexane as eluent. C.sub.21H.sub.26FN.sub.5O.sub.2 MS
m/e=400 (M+H).
EXAMPLE 71
Synthesis of 3-{3-Cyano-6-[(2,2-dimethyl-propyl)-amino
]-5-fluoro-pyridin-2-ylamino}-N-methoxy-4-methyl-benzamide
[0300] 118
(a) Synthesis of
2-Chloro-6-[(2,2-dimethyl-propyl)-amino]-5-fluoro-nicotin-
onitrile
[0301] 119
[0302] A mixture of 2,6-dichloro-3-cyano-5-fluoropyridine (1.0 g,
5.23 mmol), neopentylamine (530 mg, 6.0 mmol) and triethylamine (1
mL) in acetonitrile (20 mL) was stirred at room temp for 4 h. After
the solvent was removed under reduced pressure, the residue was
partitioned between ethyl acetate and water. The organic layer was
dried (sodium sulfate) and concentrated under reduced pressure to
afford the product (1.11 g, 87%). C.sub.11H.sub.13ClFN.sub.3 MS
m/e=242 (M+H).
(b) Synthesis of
3-{3-Cyano-6-[(2,2-dimethyl-propyl)-amino]-5-fluoro-pyrid-
in-2-ylamino}-N-methoxy-4-methyl-benzamide
[0303] 120
[0304] A mixture of
2-chloro-6-[(2,2-dimethyl-propyl)-amino]-5-fluoro-nico- tinonitrile
(150 mg, 0.62 mmol), 3-amino-N-methoxy-4-methyl-benzamide (150 mg,
0.83 mmol) and potassium fluoride (30 mg, 0.51 mmol) in DMSO (1 mL)
was heated to 150.degree. C. overnight. The reaction mixture was
allowed to cool to room temp and then partitioned between water and
ethyl acetate. The organic layer was dried (sodium sulfate) and
concentrated under reduced pressure. The product (2.5 mg, 1%) was
obtained after purification by silica gel chromatography with 30%
EtOAc in hexane as eluent. C.sub.20H.sub.24FN.sub.5O.sub.2 MS
m/e=386 (M+H)
EXAMPLE 73
Synthesis of
3-[5-Cyano-6-(2,2-dimethyl-propylamino)-3-fluoro-pyridin-2-yl-
amino]-N-methoxy-4-methyl-benzamide
[0305] 121
(a)
3-(3-Cyano-6-chloro-5-fluoro-pyridin-2-ylamino)-N-methoxy-4-methyl-ben-
zamide
[0306] 122
[0307] A mixture of 2,6-dichloro-5-fluoro-nicotinonitrile (830 mg,
4.34 mmol), 3-amino-N-methoxy-4-methyl-benzamide (576 mg, 3.2 mmol)
and triethylamine (0.5 mL) in acetonitrile (10 mL) was heated to
70.degree. C. overnight. Then the solvent was removed under reduced
pressure and the reaction mixture was partitioned between water and
ethyl acetate. The organic layer was dried (sodium sulfate) and
concentrated under reduced pressure. The product (220 mg, 21%) was
isolated after purification by silica gel chromatography.
C.sub.15H.sub.12ClFN.sub.4O.sub.2 MS m/e=335 (M+H).
(b) Synthesis of
3-[5-Cyano-6-(2,2-dimethyl-propylamino)-3-fluoro-pyridin--
2-ylamino]-N-methoxy-4-methyl-benzamide
[0308] 123
[0309] A mixture of
3-(6-chloro-5-cyano-3-fluoro-pyridin-2-ylamino)-N-meth-
oxy-4-methyl-benzamide (52 mg, 0.15 mmol), neopentylamine (0.12 mL)
and potassium fluoride (12 mg) in DMSO (1mL) was heated to
150.degree. C. overnight. The product (1.1 mg, 1.8%) was isolated
after purification by HPLC. C.sub.20H.sub.24FN.sub.5O.sub.2 MS
m/e=386 (M+H)
EXAMPLE 72
Synthesis of
3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-3-fluoro-py-
ridin-2-ylamino}-N-methoxy-4-methyl-benzamide
[0310] 124
(a)
3-(3-Cyano-6-chloro-5-fluoro-pyridin-2-ylamino)-N-methoxy-4-methyl-ben-
zamide
[0311] 125
[0312] A mixture of 2,6-dichloro-5-fluoro-nicotinonitrile (830 mg,
4.34 mmol), 3-amino-N-methoxy-4-methyl-benzamide (576 mg, 3.2 mmol)
and triethylamine (0.5 mL) in acetonitrile (10 mL) was heated to
70.degree. C. overnight. Then the solvent was removed under reduced
pressure and the reaction mixture was partitioned between water and
ethyl acetate. The organic layer was dried (sodium sulfate) and
concentrated under reduced pressure. The product (220 mg, 21%) was
isolated after purification by silica gel chromatography.
C.sub.15H.sub.12ClFN.sub.4O.sub.2 MS m/e=335 (M+H).
(b) Synthesis of
3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-3-fluor-
o-pyridin-2-ylamino}-N-methoxy-4-methyl-benzamide
[0313] 126
[0314] A mixture of
3-(6-chloro-5-cyano-3-fluoro-pyridin-2-ylamino)-N-meth-
oxy-4-methyl-benzamide (55.6 mg, 0.166 mmol),
N-methyl-neopentylamine (70 mg, 0.511 mmol), diisopropylethylamine
(0.1 mL) and potassium fluoride (12 mg) in DMSO (1 mL) was heated
to 150.degree. C. overnight. The product (3.9 mg, 5.9%) was
isolated after purification by HPLC.
C.sub.21H.sub.26FN.sub.5O.sub.2 MS m/e=400 (M+H)
EXAMPLE 52
[0315] 127
Synthesis of
3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylsul-
fanyl-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide
[0316] This compound was prepared according to procedure for the
synthesis of
3-(6-Chloro-5-cyano-2-methylsulfanyl-pyrimidin-4-ylamino)-4,N-dimethyl-
-benzamide. MS (m/z): 413 (M+H).
EXAMPLE 21
Synthesis of
3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methanesu-
lfonyl-pyrimidin-4-ylamino }-4,N-dimethyl-benzamide
[0317] 128
[0318] To a solution of
3-{5-cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]--
2-methylsulfanyl-pyrimidin-4-ylamino }-4,N-dimethyl-benzamide (0.20
g, 0.48 mmol) in acetic acid (8 mL) was added a solution of
potassium permagnate (87 mg, 0.55 mmol) in water (10 mL). The
resulting mixture was heated to 50.degree. C. for 10 minutes. The
reaction mixture was then diluted with water (20 mL), and the
product was extracted with ethyl acetate. The product was obtained
(199 mg) after drying and removing the solvent. MS (m/z): 445
(M+H).
EXAMPLE 40
Synthesis of
3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylami-
no-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide
[0319] 129
[0320]
3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methanesulfonyl-
-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide (44 mg, 0.1 mmol) and
methylamine (1 mL, 1 M in THF) were mixed in p-dioxane (1 mL) in a
sealed tube. The mixture was heated to 80.degree. C. for overnight.
The solvent was removed in vacuo, and the product (23 mg) was
purified by the silica gel column chromatography. MS (m/z): 396
(M+H).
EXAMPLE 36
Synthesis of
3-[3-Cyano-2-[(2,2-dimethyl-propyl)-methyl-amino]6-(1-methyl--
piperidin-4-yloxy)pyridin-4-ylamino]-4,N-dimethyl-benzamide
[0321] 130
[0322] To a portion of 576 mg of 1-methylpiperidine-4-ol (576 mg; 5
mmol) is added 616 mg of potassium tert.-butoxide (5.5 mmol)
followed by 4.0 mL of DMSO. After stirring this mixture at r.t. for
1 h a portion of 1.0 mL of this mixture is added at r.t. to 19 mg
of 3-{5-cyano-6-[(2,2-dimethyl--
propyl)-methyl-amino]-2-methanesulfonyl-pyrimidin-4-ylamino}-4,N-dimethyl--
benzamide (0.043 mmol) on 0.2 mL of DMSO. The mixture was heated at
60.degree. C. for 3 h. At r.t. 5 mL of ethyl acetate is added and
the organic layer is washed with brine (1.times.4 mL). The organic
layer is dried (MgSO.sub.4), volatiles are removed in vacuo and the
product is purified via reversed phase prep. HPLC. (Yield: 18.6 mg;
0.026 mmol; 56 %). MS (m/z): 480 (M+H).
EXAMPLE 41
Synthesis of
3-[5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-(4-methyl-
-[1,4]diazepan-1-yl)-pyrimidin-4-ylamino]-4,N-dimethyl-benzamide
[0323] 131
[0324]
3-[5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-(4-methyl-[1,4]-
diazepan-1-yl)-pyrimidin-4-ylamino]-4,N-dimethyl-benzamide was
synthesized similar to the synthesis of
3-{5-cyano-6-[(2,2-dimethyl-propyl)-methyl-am-
ino]-2-methylamino-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide. MS
(m/z): 479 (M+H).
EXAMPLE 42
Synthesis of
3-[5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-(4-methyl-
-[1,4]diazepan-1-yl)-pyrimidin-4-ylamino]-N-methoxy-4-methyl-benzamide
[0325] 132
(a)
3-(6-Chloro-5-cyano-2-methylsulfanyl-pyrimidin-4-ylamino)-N-methoxy-4--
methyl-benzamide
[0326] 133
[0327] 4,6-Dichloro-2-methylsulfanyl-pyrimidine-5-carbonitrile
(2.19 g, 10 mmol), 3-amino-N-methoxy-4-methyl-benzamide (1.80 g, 10
mmol) and DIEA (1.9 mL) in THF (80 mL) were stirred at room
temperature for overnight. The solvent was removed in vacuo and the
product (3.33 g, 92%) was obtained after purification by silica gel
column chromatography.
(b) Synthesis of
3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methy-
lsulfanyl-pyrimidin-4-ylamino}-N-methoxy-4-methyl-benzamide
[0328] 134
[0329]
3-(6-Chloro-5-cyano-2-methylsulfanyl-pyrimidin-4-ylamino)-N-methoxy-
-4-methyl-benzamide (3.33 g, 9.2 mmol), N-methyl-neopentylamine
hydrochloride (2.05 g, 15 mmol) and DIEA (3.87 g, 30 mmol) in THF
(10 mL) were heated to 60.degree. C. for overnight. The solvent was
removed in vacuo and the product (1.75 g) was obtained after
purification by silica gel column chromatography.
(c) Synthesis of
3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-metha-
nesulfinyl-pyrimidin-4-ylamino}-N-methoxy-4-methyl-benzamide
[0330] 135
[0331] To a solution of
3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]--
2-methylsulfanyl-pyrimidin-4-ylamino}-N-methoxy-4-methyl-benzamide
(0.10 g) in ethanol (5 mL) was added the solution of sodium
periodate (0.2 g) in water (1 mL). The resulting solution was
heated to 70.degree. C. for overnight. The solvent was evaporated
and the residue was partitioned between water and ethyl acetate.
The organic layer was separated, and concentrated, and the residue
was purified by silica gel column chromatography to afford the
sulfoxide product (90 mg). MS (m/z): 445 (M+H).
(d) Synthesis of
3-[5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-(4-me-
thyl-[1,4]diazepan-1-yl)-pyrimidin-4-ylamino]-N-methoxy-4-methyl-benzamide
[0332] 136
[0333]
3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methanesulfinyl-
-pyrimidin-4-ylamino}-N-methoxy-4-methyl-benzamide (40 mg) and
1-methyl-homopiperazine (0.05 mL) in THF (0.5 mL) were heated in
sealed tube at 75.degree. C. for overnight. After the solvent was
removed in vacuo, the residue was purified by silica gel column
chromatography to afford the product (6.8 mg). MS (m/z): 495
(M+H).
EXAMPLE 32
Synthesis of
3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylami-
no-pyrimidin-4-ylamino}-N-methoxy-4-methyl-benzamide
[0334] 137
[0335]
3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methanesulfinyl-
-pyrimidin-4-ylamino}-N-methoxy-4-methyl-benzamide (33 mg) and
methylamine (2.5 mL, 2 M in THF) were heated in a sealed tube at
75.degree. C. for overnight. After the solvent was removed in
vacuo, the residue was purified by silica gel column chromatography
to afford the product (7.3 mg). MS (m/z): 412 (M+H).
EXAMPLE 33
Synthesis of
3-[5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-(pyrrolid-
in-3-ylamino)-pyrimidin-4-ylamino]-N-methoxy-4-methyl-benzamide
[0336] 138
[0337]
3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methanesulfinyl-
-pyrimidin-4-ylamino}-N-methoxy-4-methyl-benzamide (33 mg) and
1-N-Boc-3-(R)-aminopyrrolidine (30 mg), DIEA (0.2 mL) and p-dioxane
(2 mL) were heated in a sealed tube at 75.degree. C. for overnight.
After the removal of the solvent in vacuo, the product was purified
by silica gel column chromatography and treated with TFA/DCM (1:1)
in order to remove the Boc-group. The product was then converted to
hydrochloride salt by treating it with hydrochloric acid (1 M in
ether) (Yield: 20 mg). MS (m/z) : 567 (M+H)
EXAMPLE 24
Synthesis of
3-{5-Amino-6-[(2,2-dimethyl-propyl)-methyl-amino]-pyrimidin-4-
-ylamino}-4,N-dimethyl-benzamide
[0338] 139
[0339]
3-{6-[(2,2-Dimethyl-propyl)-methyl-amino]-5-nitro-pyrimidin-4-ylami-
no}-4,N-dimethyl-benzamide (0.31 g, 0.8 mmol)) was hydrogenated
under 40 psi of hydrogen pressure in the presence of 10% Pd/C for 4
hours in Parr instrument. The catalyst was filtered off through
celite and the filtrate was concentrated to afford the product
(0.24 g, yield 84%). MS (m/z): 357 (M+H).
EXAMPLE 25
Synthesis of
3-{5-(Acetylamino)-6-[(2,2-dimethyl-propyl)-methyl-amino]-pyr-
imidin-4-ylamino}-4,N-dimethyl-benzamide
[0340] 140
[0341]
3-{5-Amino-6-[(2,2-dimethyl-propyl)-methyl-amino]-pyrimidin-4-ylami-
no}-4,N-dimethyl-benzamide (16 mg), acetyl chloride (3.5 mg),
triethylamine (0.02 ml) were stirred in methylene chloride (0.2 mL)
at room temperature for overnight. The product (2.8 mg) was
purified by preparative thin layer chromatography. MS (m/z): 399
(M+H).
EXAMPLE 14
Synthesis of
3-{5-Bromo-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylsul-
fanyl-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide
[0342] 141
[0343] To a solution of
3-{6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methyl-
sulfanyl-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide (0.10 g) in
methylene chloride (2 mL), was added sat. aq. sodium bicarbonate
(0.05 mL) and bromine (0.013 mL). The resulting mixture was stirred
at room temperature for 30 minutes, and ethyl acetate (30 mL) and
magnesium sulfate (1 g) was added. After filtration and
concentration, the residue was purified by silica gel column
chromatography to afford the product (61.9 mg). MS (m/z): 466
(M+H).
EXAMPLE 15
Synthesis of
3-{5-Bromo-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylami-
no-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide
[0344] 142
[0345]
3-{6-[(2,2-Dimethyl-propyl)-methyl-amino]-2-methylamino-pyrimidin-4-
-ylamino}-4,N-dimethyl-benzamide (33 mg), aq. sat. sodium
bicarbonate (0.05 mL) and bromine (14 mg) were stirred in methylene
chloride (1 mL) at room temperature for 4 hours. The product (14
mg) was purified with preparative thin layer chromatography. MS
(m/z): 449 (M+1).
EXAMPLE 11
Synthesis of
3-[5-Bromo-6-(2,2-dimethyl-propylamino)-2-methylamino-pyrimid-
in-4-ylamino]-4,N-dimethyl-benzamide
[0346] 143
[0347]
3-[6-(2,2-Dimethyl-propylamino)-2-methylamino-pyrimidin-4-ylamino]--
4,N-dimethyl-benzamide (35 mg), aq. sat. sodium bicarbonate (0.05
mL) and bromine (21 mg) were stirred in methylene chloride (1 mL)
at room temperature for overnight. The product (4 mg) was purified
with preparative thin layer chromatography. MS (m/z): 435 (M+H)
EXAMPLE 49
Synthesis of
3-{5-Bromo-6-[(2,2-dimethyl-propyl)-amino]-pyrimidin-4-ylamin- o}-4
N-dimethyl-benzamide
[0348] 144
(a) 3-(6-Chloro-pyrimidin-4-ylamino)-4,N-dimethyl-benzamide
[0349] 145
[0350] To a solution of 4,6-dichloropyrimidine (2.0 g, 13.4 mmol)
in p-dioxane (50 mL), 3-amino-4,N-dimethyl-benzamide (3.0 g, 18.3
mmol) and DIEA (2.3 mL) were added. The resulting mixture was
heated to reflux for 4 days. The solvent was removed in vacuo, the
residue was taken in water and ethyl acetate, and the ethyl acetate
layer was separated and concentrated. The product was purified by
silica gel column chromatography using ethyl acetate:hexane (1:1)
as eluent to afford the pale white solid product (1.6 g, yield
43%). MS (m/z): 277 (M+H).
(b) Synthesis of
3-[6-(2,2-Dimethyl-propylamino)-pyrimidin-4-ylamino]-4,N--
dimethyl-benzamide
[0351] 146
[0352] To a solution of
3-(6-chloro-pyrimidin-4-ylamino)-4,N-dimethyl-benz- amide (0.4 g,
1.44 mmol) in DMSO (3 mL), neopentylamine (0.4 mL, 3.39 mmol) was
added. The resulting solution was heated at 110.degree. C. for 4
days. The product was purified by silica gel column chromatography
using ethyl acetate as eluent to afford the product (0.45 g, yield
99%). MS (m/z): 328 (M+H).
(c) Synthesis of
3-{5-Bromo-6-[(2,2-dimethyl-propyl)-amino]-pyrimidin-4-yl-
amino}-4,N-dimethyl-benzamide
[0353] 147
[0354] To a solution of
3-{6-[(2,2-dimethyl-propyl)-amino]-pyrimidin-4-yla-
mino}-4,N-dimethyl-benzamide (0.45 g, 1.37 mmol) in methylene
chloride (10 mL), was added sat. aq. sodium bicarbonate (2 mL) and
bromine (0.07 mL, 1.37 mmol). The resulting mixture was stirred at
room temperature for 18 hours, and the reaction was then diluted
with water (20 mL). The organic layer was separated and the aqueous
layer was extracted with DCM (10 mL). The combined organic layer
was dried (sodium sulfate), filtered and concentrated. The residue
was purified by silica gel column chromatography using 2% methanol
in DCM as eluent to afford the product (314 mg, yield 56%). MS
(m/z): 406 (M+H).
EXAMPLE 2
Synthesis of
3-{5-Bromo-6-[(2,2-dimethyl-propyl)-methyl-amino]-pyrimidin-4-
-ylamino}-4,N-dimethyl-benzamide
[0355] 148
(a) 3-(6-Chloro-pyrimidin-4-ylamino)-4,N-dimethyl-benzamide
[0356] 149
[0357] To a solution of 4,6-dichloropyrimidine (2.0 g, 13.4 mmol)
in p-dioxane (50 mL), 3-amino-4,N-dimethyl-benzamide (3.0 g, 18.3
mmol) and DIEA (2.3 mL) were added. The resulting mixture was
heated to reflux for 4 days. The solvent was removed in vacuo, the
residue was taken in water and ethyl acetate, and the ethyl acetate
layer was separated and concentrated. The product was purified by
silica gel column chromatography using ethyl acetate:hexane (1:1)
as eluent to afford the pale white solid product (1.6 g, yield
43%). MS (m/z): 277 (M+H).
(b) Synthesis of
3-{6-[(2,2-Dimethyl-propyl)-methyl-amino]-pyrimidin-4-yla-
mino}-4,N-dimethyl-benzamide
[0358] 150
[0359] To a solution of
3-(6-chloro-pyrimidin-4-ylamino)-4,N-dimethyl-benz- amide (0.4 g,
1.44 mmol) in DMSO (3 mL), N-methylneopentylamine hydrochloride
(0.4 g, 2.9 mmol) and DIEA (0.5 mL, 2.9 mmol) were added. The
resulting solution was heated at 110.degree.0 C. for 4 days. The
product was purified by silica gel column chromatography using
ethyl acetate as eluent to afford the product (0.46 g, yield 99%).
MS (m/z): 342 (M+H).
(c) Synthesis of
3-{5-Bromo-6-[(2,2-dimethyl-propyl)-methyl-amino]-pyrimid-
in-4-ylamino}-4,N-dimethyl-benzamide
[0360] 151
[0361] To a solution of
3-{6-[(2,2-dimethyl-propyl)-methyl-amino]-pyrimidi-
n-4-ylamino}-4,N-dimethyl-benzamide (0.46 g, 1.34 mmol) in
methylene chloride (10 mL), was added sat. aq. sodium bicarbonate
(2 mL) and bromine (0.07 mL, 1.37 mmol). The resulting mixture was
stirred at room temperature for 18 hours, and the reaction was then
diluted with water (20 mL). The organic layer was separated and the
aqueous layer was extracted with DCM (10 mL). The combined organic
layer was dried (sodium sulfate), filtered and concentrated. The
residue was purified by silica gel column chromatography using 2%
methanol in DCM as eluent to afford the product (250 mg, yield
44%). MS (m/z): 420 (M+H).
EXAMPLE 74
Synthesis of
3-(R)-[5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-(pyrr-
olidin-3-ylamino)-pyrimidin-4-ylamino]-4-N-dimethyl-benzamide
[0362] 152
(a)
3-(R)-[4-[(2,2-Dimethyl-propyl)-methyl-amino]-6-(2-methyl-5-methylcarb-
amoyl-phenylamino)-pyrimidin-2-ylamino]-pyrrolidine-1-carboxylic
acid tert-butyl ester
[0363] 153
[0364] A mixture of
3-(R)-[4-fluoro-6-(2-methyl-5-methylcarbamoyl-phenylam-
ino)-pyrimidin-2-ylamino]-pyrrolidine-1-carboxylic acid tert-butyl
ester (0.36 g, 0.81 mmol), N-methyl-neopentylamine hydrochloride
(411 mg, 3 mmol) and DIEA(0.4 mL) in 1,4-dioxane (0.5 mL) was
stirred at 90.degree. C. overnight. After removing the solvent
under reduced pressure the desired product (81 mg) was purified by
silica gel chromatography. C.sub.28H.sub.43N.sub.7O.sub.3 MS
m/e=526 (M+H).
(b) Synthesis of
3-(R)-[5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-(-
pyrrolidin-3-ylamino)-pyrimidin-4-
[0365] 154
[0366] To a mixture of
3-(R)-[4-[(2,2-dimethyl-propyl)-methyl-amino]-6-(2--
methyl-5-methylcarbamoyl-phenylamino)-pyrimidin-2-ylamino]-pyrrolidine-1-c-
arboxylic acid tert-butyl ester (70 mg, 0.0001 mmol) and sodium
cyanide (0.044 g, 0.89 mmol) in sat. aq. sodium bicarbonate (1 mL)
and methylene chloride (3 mL) at room temp was added bromine (0.045
mL, 0.87 mmol). The resulting mixture was stirred for 16 h at room
temp, then diluted with water and extracted with methylene chloride
(2.times.15 mL). The combined organic layer was dried (sodium
sulfate), filtered and concentrated under reduced pressure. This
product was then treated with a mixture of trifluoroacetic acid and
methylene chloride (1:1 v/v, 1 mL). The resulting solution was
stirred at room temp for 2 h, then the solvent was removed under
reduced pressure and the product was purified by HPLC.
C.sub.24H.sub.34N.sub.8O MS m/e=451 (M+H).
EXAMPLE 75
Synthesis of
3-(R)-[5-Cyano-4-[(2,2-dimethyl-propyl)-methyl-amino]-6-(pyrr-
olidin-3-ylamino)-pyrimidin-2-ylamino]-4-N-dimethyl-benzamide
[0367] 155
[0368] To a mixture of
3-(R)-[6-[(2,2-dimethyl-propyl)-methyl-amino]-2-(2--
methyl-5-methylcarbamoyl-phenylamino)-pyrimidin-4-ylamino]-pyrrolidine-1-c-
arboxylic acid tert-butyl ester (128 mg, 0.0002 mmol) and sodium
cyanide (0.044 g, 0.89 mmol) in sat. aq. sodium bicarbonate (1 mL)
and methylene chloride (3 mL) was added bromine (0.045 mL, 0.87
mmol). The resulting mixture was continued stirring at room temp
for 16 h, then diluted with water and extracted with methylene
chloride (2.times.15 mL). The combined organic layer was dried
(sodium sulfate), filtered and concentrated under reduced pressure.
This product was then treated with a mixture of trifluoroacetic
acid and methylene chloride (1:1 v/v, 1 mL). The resulting solution
was stirred at room temp for 2 h. The solvent was removed under
reduced pressure, and the product was purified by HPLC.
C.sub.24H.sub.34N.sub.8O MS m/e=451 (M+H).
EXAMPLE 62
Synthesis of
3-[5-Cyano-4-[(2,2-dimethyl-propyl)-methyl-amino]-6-(4-methyl-
-[1,4]diazepan-1-yl)-pyrimidin-2-ylamino]-N-methoxy-4-methyl-benzamide
[0369] 156
[0370] To a stirred mixture of
3-[4-[(2,2-dimethyl-propyl)-methyl-amino]-6-
-(4-methyl-[1,4]diazepan-1-yl)-pyrimidin-2-ylamino]-N-methoxy-4-methyl-ben-
zamide (140 mg, 0.29 mmol) and sodium cyanide (0.044 g, 0.89 mmol)
in sat. aq. sodium bicarbonate (1 mL) and methylene chloride (3 mL)
was added bromine (0.045 mL, 0.87 mmol). The resulting mixture was
stirred for 16 h at room temp, then diluted with water and
extracted with methylene chloride (2.times.15 mL). The combined
organic layer was dried (sodium sulfate), filtered and concentrated
under reduced pressure, and the product was purified by HPLC.
C.sub.26H.sub.38N.sub.8O.sub.2 MS m/e=495 (M+H). Deprotection was
performed as described in Example 75.
EXAMPLE 61
Synthesis of
3-[5-Cyano-2-[(2,2-dimethyl-propyl)-methyl-amino]-6-(4-methyl-
-[1,4]diazepan-1-yl)-pyrimidin-4-ylamino]-N-ethoxy-4-methyl-benzamide
[0371] 157
[0372] To a stirred mixture of
3-[2-[(2,2-dimethyl-propyl)-methyl-amino]-6-
-(4-methyl-[1,4]diazepan-1-yl)-pyrimidin-4-ylamino]-N-ethoxy-4-methyl-benz-
amide (50 mg, 0.1 mmol) and sodium cyanide (0.044 g, 0.89 mmol) in
sat. aq. sodium bicarbonate (1 mL) and methylene chloride (3 mL)
was added at room temp bromine (0.045 mL, 0.87 mmol). The resulting
mixture was continued stirring for 16 h at room temp. The reaction
mixture was diluted with water and extracted with methylene
chloride (2.times.15 mL). The combined organic layer was dried
(sodium sulfate), filtered and concentrated under reduced pressure,
and the product was purified by HPLC.
C.sub.27H.sub.40N.sub.8O.sub.2 MS m/e=509 (M+H).
EXAMPLE 43
Synthesis of
4-[(2,2-Dimethyl-propyl)-methyl-amino]-6-[2-methyl-5-(4H-[1,2-
,4]triazol-3-yl)-phenylamino]-2-(pyrrolidin-3(R)-ylamino)-pyrimidine-5-car-
bonitrile
[0373] 158
[0374] To a solution of
2-methyl-5-(4H-[1,2,4]triazol-3-yl)-phenylamine (174 mg, 1 mmol) in
4 mL of DMF was added 0.17 mL of diisopropylethylamine (1 mmol) and
4,6-dichloro-2-methylsulfanyl-pyrimidi- ne-5-carbonitrile (219 mg,
1 mmol). The resulting solution was stirred at room temperature
overnight, then partitioned between ethyl acetate and water. The
organic layer was washed with water, brine and dried over
MgSO.sub.4. Removal of volatiles in vacuo and purification by flash
chromatography gave 78 mg of the product (Yield: 22%). MS (m/z)
calcd for C.sub.15H.sub.12ClN.sub.7S (MH+), 358.1, found,
358.3.
[0375] (b) 159
[0376] To a solution of
4-chloro-2-methylsulfanyl-6-[2-methyl-5-(4H-[1,2,4-
]triazol-3-yl)-phenylamino]-pyrimidine-5-carbonitrile (71 mg, 0.2
mmol ) in 2 mL of n-BuOH was added 137 mg of
N-(2,2-dimethylpropyl)methyl amine HCl salt (1 mmol) and 0.17 mL of
diisopropylethylamine (1 mmol) and the resulting solution was
stirred at 600.degree. C. for 24 h. Removal of volatiles in vacuo
and purification by flash chromatography gave 68 mg of the product
(Yield: 81%). MS (m/z) calcd for C.sub.21H.sub.26N.sub.8S (MH+),
423.2, found, 423.3. 160
[0377] To a solution of
4-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylsul-
fanyl-6-[2-methyl-5-(4H-[1,2,4]triazol-3-yl)-phenylamino]-pyrimidine-5-car-
bonitrile (76 mg, 0.18 mmol) in 3 mL of acetic acid was added 0.5
mL water followed by 50 mg of potassium permanganate (0.36 mmol).
This solution was stirred at 60.degree. C. for 10 minutes, then
diluted with water and extracted with ethyl acetate. The organic
layer was washed with water, brine and dried over MgSO.sub.4.
Removal of volatiles in vacuo gave 58 mg of the crude product which
was used in the next step without further purification (Yield:
71%). MS (m/z) calcd for C.sub.21H.sub.26N.sub.8O.su- b.2S (MH+),
455.2, found, 455.3. 161
[0378] A solution of
4-[(2,2-dimethyl-propyl)-methyl-amino]-2-methanesulfo-
nyl-6-[2-methyl-5-(4H-[1,2,4]triazol-3-yl)-phenylamino]-pyrimidine-5-carbo-
nitrile (10 mg, 0.022 mmol) and
1-tert-butoxycarbony-3(R)-amino-pyrrolidin- e (0.08 g, 0.4 mmol) in
2 mL of acetonitrile was heated with stirring at 80.degree. C. for
18 h. Volatiles were the removed in vacuo and the product was
purified by flash chromatography.
[0379] This purified product was then dissolved in 3 mL of a
solution of 50% TFA in CH.sub.2Cl.sub.2 (v/v) and stirred at room
temp for 30 min. Removal of volatiles in vacuo and purification via
prep. HPLC gave the product as TFA salt. The purified product was
then dissolved in 1N HCl (g) in diethyl ether and evaporated to
give 0.8 mg of the final product. (Yield: 8%). MS (m/z) calcd for
C.sub.24H.sub.32N.sub.10 (MH+), 461.3, found, 461.4.
EXAMPLE 17
Synthesis of
4-[(2,2-Dimethyl-propyl)-methyl-amino]-6-[2-methyl-5-(4H-[1,2-
,4]triazol-3-yl)-phenylamino]-pyrimidine-5-carbonitrile
[0380] 162
[0381] To a solution of
4-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylsul-
fanyl-6-[2-methyl-5-(4H-[1,2,4]triazol-3-yl)-phenylamino]-pyrimidine-5-car-
bonitrile (60 mg, 0.142 mmol) in 3 mL of 50% ethanol/water (v/v)
was added 0.4 mL of 50% Raney-Ni in water solution. The solution
was refluxed under argon for 18 hours. The solution was evaporated
under vacuum. The resulting residue was purified by flash
chromatography to afford 5.6 mg of the final product (Yield: 10%).
MS (m/z) calcd for C.sub.20H.sub.24N.sub.8 (MH+), 377.2, found,
377.4.
EXAMPLE 80
Synthesis of
3-{5-Bromo-2-[2,2-dimethyl-propyl)-methyl-amino]-pyrimidin-4--
ylamino}-4,N-dimethyl-benzamide
[0382] 163
(a)
3-(5-Bromo-2-chloro-pyrimidin-4-ylamino)-4,N-dimethyl-benzamide
[0383] 164
[0384] To a solution of 0.38 mL of 5-Bromo-2,4-dichloror-pyrimidine
(70 mg; 3.1 mmol) in 5 mL of THF at 0.degree. C. is added dropwise
a solution of 0.644 mL of N,N-diisopropylethylamine (478 mg; 3.7
mmol) and of 506 mg of 3-Amino-4,N-dimethyl-benzamide ((3.1 mmol)
in 2 mL of THF. This solution is continued stirring at 0.degree.0
C. for 1 h, then at 25.degree. C. for 30 min. After removal of
volatiles in vacuo the product was purified via silica gel
chromatography (20% ethyl acetate in hexanes) to yield 304 mg of a
white powder (0.85 mmol; 25% yield). MS (m/z): 355 (M+H).
(b) Synthesis of
3-{5-Bromo-2-[2,2-dimethyl-propyl)-methyl-amino]-pyrimidi-
n-4-ylamino}-4,1N-dimethyl-benzamide
[0385] 165
[0386] A mixture of 51 mg of
3-(5-Bromo-2-chloro-pyrimidin-4-ylamino)-4,N-- dimethyl-benzamide
(0.143 mmol), 59 mg of (2,2-dimethylpropyl)-methylamine- ,
hydrochloride (0.430 mmol) and 0.112 mL of
N,N-diisopropylethylamine (83 mg; 0.645 mmol) is heated to
120.degree. C. for 18 h, then allowed to cool to r.t. 5 ml of ethyl
acetate are added and the organic layer is washed with brine
(1.times.5 ml). The aqueous layer is extracted with ethyl acetate
(3.times.5 ml) and the combined organic layers are dried
(MgSO.sub.4). After removal of volatiles in vacuo the product was
purified via silica gel chromatography (20% ethyl acetate in
hexanes) to yield 41 mg of an colorless oil (0.097 mmol; yield:
68%). MS (m/z) : 420 (M+H)
EXAMPLE 4
Synthesis of
3-[5-Bromo-2-(pyrrolidin-3(R)-(ylamino)-pyrimidin-4-ylamino]--
4,N-dimethyl-benzamide
[0387] 166
[0388] A solution of 22 mg of
3-(5-bromo-2-chloro-pyrimidin-4-ylamino)-4,N- -dimethyl-benzamide
(0.06 mmol) and 46 mg of 3-amino-pyrrolidine-1-carboxy- lic acid
tert.-butyl ester (0.247 mmol) is heated at 120.degree. C. in 0.3
mL DMSO for 3 d. After addition of 5 mL of ethyl acetate at r.t.
the organic layer is washed with a sat. solution of NaHCO.sub.3 in
water (3.times.4 mL). The organic layer is dried (MgSO.sub.4),
volatiles are removed in vacuo and the product is purified via
prep. HPLC. (Yield: 5.7 mg; 0.013 mmol; 22%) MS (m/z) : 444
(M.sup.+) ; 455 (M+Na)
[0389] The purified product is dissolved in 1 mL of MeOH and 3 mL
of a 1N solution of HCl in diethyl ether is added. The resulting
solution is stirred at r.t for 30 min, then volatiles are removed
in vacuo and the product is purified via prep. HPLC. (Yield: 4.3
mg; 0.0002 mmol; 0.4%). MS (m/z): 405 (M+H).
EXAMPLE 3
Synthesis of
3-[Bromo-2-(1-methyl-piperidin-4-yloxy)-pyrimidin-4-ylamino]4-
,N-dimethylbenzamide
[0390] 167
[0391] To a portion of 576 mg of 1-methylpiperidine-4-ol (576 mg; 5
mmol) is added 616 mg of potassium tert.-butoxide (5.5 mmol)
followed by 4.0 mL of DMSO. After stirring this mixture at r.t. for
1 h a portion of 1.0 mL of this mixture is added at r.t. to 21 mg
of 3-(5-Bromo-2-chloro-pyrimidi-
n-4-ylamino)-4,N-dimethyl-benzamide (0.06 mmol). The resulting
mixture is heated at 120.degree. C. for 18 h and then allowed to
cool to r.t. A portion of 5 mL of ethyl acetate is added and the
resulting solution is washed with a sat. solution of NaHCO3 in
water (3.times.4 mL). The organic layer is dried (MgSO.sub.4),
volatiles are removed in vacuo and the product is purified via
prep. HPLC. (Yield: 5.7 mg; 0.013 mmol; 22%). MS (m/z): 444
(M.sup.+); 455 (M+Na).
EXAMPLE 53
Synthesis of
3-{5-Cyano-2-[2,2-dimethyl-propyl)-methyl-amino]-pyrimidin-4--
ylamino}-4,N-dimethyl-benzamide
[0392] 168
[0393] A mixture of 15 mg of
3-{5-Bromo-2-[2,2-dimethyl-propyl)-methyl-ami-
no]-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide (0.036 mmol) and
100 mg of CuCN (1.12 mmol) in 1.0 mL of 1-methyl-2-pyrrolidinone is
heated to 140.degree. C. for 18 h. The mixture was allowed to cool
to r.t. and 2.0 mL of MeOH were added. After removing the
precipitate by filtration volatiles were evaporated and the product
was puriefied by prep. HPCL. Yield: 1.7 mg (0.005 mmol; 13%). MS
(m/z): 367 (M+H).
EXAMPLE 1
Synthesis of
3-{5-cyano-4-[-(2,2-dimethyl)(-propyl)-methyl-amino]-pyrimidi-
n-2-ylamino}-4,N-dimethyl-benzamide
[0394] 169
[0395] To a solution of 1.0 g of 2,4-dichloropyrimidine-5-carbonyl
chloride (4.73 mmol) in 5 ml of THF at 0.degree. C. is added
dropwise a 0.5 M solution of NH.sub.3 in 1,4-dioxane. The progress
of the amide formation is followed via HPLC and the solution of
NH.sub.3 in 1,4-dioxane is added until all of the acid chloride is
consumed. Then volatiles are removed in vacuo to yield a white
solid.
[0396] To the crude product from above is added at r.t. 25 mL of
POCl.sub.3 and the resulting mixture is heated to 100.degree. C.
for 4 h. Volatiles are removed in vacuo, the crude product is
absorbed on silica gel and washed off with 20% ethyl acetate in
hexane to give a white solid.
[0397] To 20 mg of the product so obtained in 0.5 mL of THF is
added at r.t. 0.022 mL of N,N-diisopropylethylamine 16.3 mg; 0.126
mmol) followed by 21 mg of 3-amino-4,N-dimethylbenzamide (0.126
mmol). The mixture is stirred at r.t. for 2 h, then 0.5 mL of THF
are added followed by 32 mg of (2,2-dimethyl-propyl)-methyl-amine,
hydrochloride (0.232 mmol) and 0.044 mL of
N,N-diisopropylethylamine (32.6 mg; 0.252 mmol). The resulting
mixture is heated at 60.degree. C. for 18 h. Volatiles are removed
in vacuo and the crude mixture containing two separable
regioisomers is purified via reversed phase prep. HPLC. (Yield: 3.9
mg: 0.011 mmol; 8%). MS (m/z): 367 (M+H).
EXAMPLE 56
Synthesis of
3-{6-[(2,2-Dimethyl-propyl)-methyl-amino]-5-nitro-pyrimidin-4-
-ylamino}-4,N-dimethyl-benzamide
[0398] 170
(a)
(6-Chloro-5-nitro-pyrimidin-4-yl)-(2,2-dimethyl-propyl)-methyl-amine
[0399] 4,6-Dichloro-5-nitro-pyrimidine (0.20 g, 1.0 mmol),
N-methyl-neopentylamine hydrochloride (0.14 g) and DIEA (0.2 mL)
were stirred in acetone (5 mL) at 0.degree. C. for 4 hours. The
solvent was removed in vacuo and the crude product was used for the
next reaction without purification.
(b) Synthesis of
3-{6-[(2,2-Dimethyl-propyl)-methyl-amino]-5-nitro-pyrimid-
in-4-ylamino}-4,N-dimethyl-benzamide
[0400] 171
[0401]
(6-Chloro-5-nitro-pyrimidin-4-yl)-(2,2-dimethyl-propyl)-methyl-amin-
e (1.0 mmol) was dissolved in p-dioxane (2 mL), and
3-amino-4,N-dimethyl-benzamide (0.2 g, 1.2 mmol) and DIEA (0.3 mL)
were added. The resulting mixture was heated to 80.degree. C. for
overnight. The product (0.31 g, yield 80%) was purified by column
silica gel chromatography. MS (m/z): 387 (M+H).
EXAMPLE 26
Synthesis of
N-(2,2-Dimethyl-propyl)-N-methyl-N'-[2-methyl-5-(4H-[1,2,4]tr-
iazol-3-yl)-phenyl]-5-nitro-pyrimidine-4,6-diamine
[0402] 172
[0403] 4,6-Dichloro-5-nitro-pyrimidine (193 mg, 1 mmol) was
dissolved in 4 mL acetone at 0.degree. C. To the solution was added
N-(2,2-dimethylpropyl)methyl amine HCl salt (137 mg, 1 mmol) and
diisopropylethylamine (0.17 mL, 1 mmol). The solution was stirred
at 0.degree. C. for 10 minutes, then room temperature for 3 hours,
and evaporated in vacuo. The crude product was purified by flash
chromatography to afford 180 mg of the product (Yield: 69%). MS
(m/z) calcd for C.sub.10H.sub.15ClN.sub.4O.sub.2 (MH+), 259.1,
found, 259.3. 173
[0404] A solution of
(6-Chloro-5-nitro-pyrimidin-4-yl)-(2,2-dimethyl-propy-
l)-methyl-amine (185 mg, 0.72 mmol), diisopropylethylamine (0.13
mL, 0.72 mmol) and 2-Methyl-5-(4H-[1,2,4]triazol-3-yl)-phenylamine
(126 mg, 0.72 mmol) in 3 mL of n-BuOH was heated with stirring at
80.degree. C. for 18 h. The solvent was then evaporated in vacuo
and the crude product was purified by flash chromatography to
afford 87 mg of the product (30%). MS (m/z) calcd for
C.sub.19H.sub.24N.sub.8O.sub.2 (MH+), 397.2, found, 397.3.
EXAMPLE 28
Synthesis of
N-(2,2-Dimethyl-propyl)-N-methyl-N"-[2-methyl-5-(4H-[1,2,4]tr-
iazol-3-yl)phenyl]-pyrimidine-4,5,6-triamine
[0405] 174
[0406] To a solution of
N-(2,2-dimethyl-propyl)-N-methyl-N'-[2-methyl-5-(4-
H-[1,2,4]triazol-3-yl)phenyl]-5-nitro-pyrimidine-4,6-diamine (20
mg, 0.05 mmol) in 5 mL of methanol was added a catalytic amount of
10% Pd/C. The vessel was placed under a hydrogen atmosphere of 20
psi for 1 h at room temperature. The solution was filtered, and the
filtrate was evaporated under vacuum to afford 4.6 mg of the
product (Yield: 25%). MS (m/z) calcd for C.sub.19H.sub.26N.sub.8
(MH+), 367.2, found, 367.4.
EXAMPLE 12
Synthesis of
3-{2-(3-Dimethylamino-propylamino)-6-[(2,2-dimethyl-propyl)-m-
ethyl-amino]-5-nitro-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide
[0407] 175
[0408] 2-Methylsulfanyl-pyrimidine-4,6-diol (1.58 g, 10 mmol) was
dissolved in 10 mL of con. H.sub.2SO.sub.4 at 0.degree. C. To the
solution was added 0.84 mL of nitric acid drop-wise. The solution
was stirred at 0.degree. C. for 30 minutes, then room temperature
for 2 h. The solution was poured into ice water. The yellowish
solid precipitated out of the solution was collected, washed with
cold water and dried to afford 400 mg of the product (Yield: 20%).
176
[0409] 2-Methylsulfanyl-5-nitro-pyrimidine-4,6-diol (203 mg, 1
mmol) was dissolved in 4 mL of phosphorus oxychloride. The solution
was refluxed at 120.degree. C. for two hours. The solution was
evaporated under vacuum. The oily residue was purified by flash
chromatography to afford 80 mg of the product (Yield: 35%). 177
[0410] 4,6-Dichloro-2-methylsulfanyl-5-nitro-pyrimidine (30 mg,
0.13 mmol) and 3-amino-4,N-dimethyl-benzamide (22 mg, 0.13 mmol)
were dissolved in 2 mL of THF. The solution was stirred at
0.degree. C. for 30 minutes, and evaporated under vacuum. The
product thus obtained was directly used for the next reaction
without purification. 178
[0411] The intermediate so obtained was dissolved in 2 mL of THF.
To the solution was added N-(2,2-dimethylpropyl)methyl amine HCl
salt (36 mg, 0.26 mmol) and diisopropylethylamine (0.05 mL, 0.26
mmol). The solution was stirred at room temperature for 1 h, and
evaporated under vacuum. The residue was purified by flash
chromatography to afford 5.5 mg of the product (Yield: 10%, two
steps). MS (m/z) calcd for C.sub.20H.sub.28N.sub.6O.sub.3S (MH+),
433.2, found, 433.2. 179
[0412]
3-{6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylsulfanyl-5-nitro--
pyrimidin-4-ylamino}-4,N-dimethyl-benzamide (24 mg, 0.06 mmol) was
dissolved in 1 mL acetic acid. To the solution was added two drops
of water and potassium permanganate (18 mg, 0.12 mmol). The
solution was stirred at 60.degree. C. for 10 minutes. The solution
was diluted with water and extracted with ethyl acetate. The
organic layer was washed with water, brine, dried over MgSO.sub.4,
and evaporated under vacuum. The residue was further purified by
flash chromatography to afford 4.2 mg of the product (Yield: 15%).
MS (m/z) calcd for C.sub.2OH.sub.28N.sub.6O.sub- .5S (MH+), 465.2,
found, 465.2.
[0413] (f) 180
[0414] A solution of
3-{6-[(2,2-Dimethyl-propyl)-methyl-amino]-2-methanesu-
lfonyl-5-nitro-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide (4.2 mg,
0.01 mmol) and 3-(Dimethylamino)propylamine (0.2 mL) in 2 mL of
acetonitrile was heated with stirring at 80.degree. C. for 16 h.
The solvent was then evaporated under vacuum and the crude product
was purified by flash chromatography to afford 2.8 mg of the
product (Yield: 64%). MS (m/z) calcd for
C.sub.24H.sub.38N.sub.8O.sub.3 (MH+), 487.3, found, 487.3.
EXAMPLE 5
Synthesis of
3-[6-[(2,2-Dimethyl-propyl)-methyl-amino]-2-(1-methyl-piperid-
in-4-yloxy)-5-nitro-pyrimidin-4-ylamino]-4,N-dimethyl-benzamide
[0415] 181
[0416] 1-Methyl-piperidin-4-ol (56 mg, 0.5 mmol) was dissolved in 2
mL of DMSO. To the solution was added potassium tert-butoxide (56
mg, 0.5 mmol). The solution was stirred at room temperature for 1
h. The solution was then added to a solution of
3-{6-[(2,2-Dimethyl-propyl)-methyl-amino]-
-2-methanesulfonyl-5-nitro-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide
(4 mg, 0.01 mmol) in 0.5 mL of DMSO. The solution was stirred at
80.degree. C. for 3 h, and extracted with ethyl acetate and water.
The organic layer was washed with water, brine, dried over MgSO4,
and evaporated under vacuum. The residue was purified by
semi-preparative hplc column to afford 0.7 mg of the product
(Yield: 16%). MS (m/z) calcd for C.sub.25H.sub.37N.sub.7O.sub.4
(MH+), 500.2, found, 500.1.
[0417] Although the present invention has been described in some
detail by way of illustration and example, for purposes of clarity
and understanding, it will be apparent that certain changes and
modifications may be practiced within the scope of the appended
claims.
* * * * *