U.S. patent application number 12/361409 was filed with the patent office on 2009-05-28 for a2b adenosine receptor antagonists.
Invention is credited to Elfatih Elzein, Rao Kalla, Xiaofen Li, Tim Marquart, Jeff Zablocki.
Application Number | 20090137802 12/361409 |
Document ID | / |
Family ID | 34633233 |
Filed Date | 2009-05-28 |
United States Patent
Application |
20090137802 |
Kind Code |
A1 |
Elzein; Elfatih ; et
al. |
May 28, 2009 |
A2B ADENOSINE RECEPTOR ANTAGONISTS
Abstract
Disclosed are processes for the synthesis of novel compounds
that are A.sub.2B adenosine receptor antagonists, useful for
treating various disease states, including asthma and diarrhea.
Inventors: |
Elzein; Elfatih; (Fremont,
CA) ; Kalla; Rao; (Sunnyvale, CA) ; Marquart;
Tim; (Santa Clara, CA) ; Zablocki; Jeff; (Los
Altos, CA) ; Li; Xiaofen; (Mountain View,
CA) |
Correspondence
Address: |
CV THERAPEUTICS, INC.
3172 PORTER DRIVE
PALO ALTO
CA
94304
US
|
Family ID: |
34633233 |
Appl. No.: |
12/361409 |
Filed: |
January 28, 2009 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
11508592 |
Aug 22, 2006 |
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12361409 |
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|
10719102 |
Nov 21, 2003 |
7125993 |
|
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11508592 |
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|
10431167 |
May 6, 2003 |
6977300 |
|
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10719102 |
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10290921 |
Nov 8, 2002 |
6825349 |
|
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10431167 |
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60348222 |
Nov 9, 2001 |
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Current U.S.
Class: |
544/310 ;
544/312 |
Current CPC
Class: |
C07D 239/545 20130101;
A61P 11/06 20180101; C07D 473/06 20130101; A61P 9/00 20180101; A61P
25/00 20180101; C07D 473/00 20130101; A61P 1/00 20180101; A61P
43/00 20180101; A61P 37/00 20180101; C07D 403/12 20130101; C07D
473/08 20130101 |
Class at
Publication: |
544/310 ;
544/312 |
International
Class: |
C07D 239/545 20060101
C07D239/545; C07D 403/12 20060101 C07D403/12 |
Claims
1. An intermediate of the formula: ##STR00067## wherein: R.sup.1
and R.sup.2 are independently chosen from methyl, ethyl, n-propyl,
2-methylpropyl, and cyclopropylmethyl.
2. The intermediate of claim 1, wherein R.sup.1 is n-propyl and
R.sub.2 is ethyl: ##STR00068## namely
6-[2-(dimethylamino)-1-azavinyl]-1-ethyl-3-propyl-1,3-dihydropyrimidine-2-
,4-dione.
3. An intermediate of the formula: ##STR00069## wherein R.sup.1 is
n-propyl or cyclopropylmethyl.
4. The intermediate of claim 3, wherein R.sup.1 is n-propyl:
##STR00070## namely
N-(6-amino-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[3-(trifluor-
omethyl)-phenyl]methyl}pyrazol-4-yl)carboxamide.
5. An intermediate of the formula: ##STR00071## wherein R.sup.2 is
methyl or ethyl.
6. The intermediate of claim 5, wherein R.sup.2 is ethyl:
##STR00072##
N-(6-amino-1-ethyl-2,4-dioxo(1,3-dihydropyrimidin-5-yl))(1-{{[3-(trifluor-
omethyl)phenyl]methyl}pyrazol-4-yl)carboxamide.
7. An intermediate of the formula: ##STR00073## wherein R.sup.1 and
R.sup.2 are independently chosen from methyl, ethyl, n-propyl,
2-methylpropyl, and cyclopropylmethyl.
Description
[0001] This application is a Divisional of U.S. patent application
Ser. No. 11/508,592, filed Aug. 22, 2006, which is a Divisional of
U.S. patent application Ser. No. 10/719,102, filed Nov. 21, 2003,
which issued Oct. 24, 2006 as U.S. Pat. No. 7,125,993, which is a
Continuation in Part of U.S. patent application Ser. No.
10/431,167, filed May 6, 2003, which issued Dec. 20, 2005, as U.S.
Pat. No. 6,977,300, which is a Continuation in Part of U.S. patent
application Ser. No. 10/290,921, filed Nov. 8, 2002, which issued
Nov. 30, 2004, as U.S. Pat. No. 6,825,349, which claims priority to
U.S. Provisional Patent Application Ser. No. 60/348,222, filed Nov.
9, 2001, the complete disclosures of which are hereby incorporated
by reference.
FIELD OF THE INVENTION
[0002] The present invention relates to A.sub.2B adenosine receptor
antagonists, and to their use in treating mammals for various
disease states, such as gastrointestinal disorders, immunological
disorders, neurological disorders, and cardiovascular diseases due
to both cellular hyperproliferation and apoptosis, and the like.
The invention also relates to methods for the preparation of such
compounds, and to pharmaceutical compositions containing them.
BACKGROUND
[0003] Adenosine is a naturally occurring nucleoside, which exerts
its biological effects by interacting with a family of adenosine
receptors characterized as A.sub.1, A.sub.2A, A.sub.2B, and
A.sub.3, all of which modulate important physiological processes.
For example, A.sub.2A adenosine receptors modulate coronary
vasodilation, A.sub.2B receptors have been implicated in mast cell
activation, asthma, vasodilation, regulation of cell growth,
intestinal function, and modulation of neurosecretion (See
Adenosine A.sub.2B Receptors as Therapeutic Targets, Drug Dev Res
45.198; Feoktistov et al., Trends Pharmacol Sci 19:148-153), and
A.sub.3 adenosine receptors modulate cell proliferation
processes.
[0004] Adenosine A.sub.2B receptors are ubiquitous, and regulate
multiple biological activities. For example, adenosine binds to
A.sub.2B receptors on endothelial cells, thereby stimulating
angiogenesis. Adenosine also regulates the growth of smooth muscle
cell populations in blood vessels. Adenosine stimulates A.sub.2B
receptors on mast cells, thus modulating Type I hypersensitivity
reactions. Adenosine also stimulates gastrosecretory activity by
ligation with A.sub.2B in the intestine.
[0005] While many of these biological effects of adenosine are
necessary to maintain normal tissue homeostasis, under certain
physiological changes it is desirable to modulate its effects. For
example, the binding of A.sub.2B receptors stimulates angiogenesis
by promoting the growth of endothelial cells. Such activity is
necessary in healing wounds, but the hyperproliferation of
endothelial cells promotes diabetic retinopathy. Also, an
undesirable increase in blood vessels occurs in neoplasia.
Accordingly, inhibition of the binding of adenosine to A.sub.2B
receptors in the endothelium will alleviate or prevent
hypervasculation, thus preventing retinopathy and inhibiting tumor
formation.
[0006] A.sub.2B receptors are found in the colon in the basolateral
domains of intestinal epithelial cells, and when acted upon by the
appropriate ligand act to increase chloride secretion, thus causing
diarrhea, which is a common and potentially fatal complication of
infectious diseases such as cholera and typhus. A.sub.2B
antagonists can therefore be used to block intestinal chloride
secretion, and are thus useful in the treatment of inflammatory
gastrointestinal tract disorders, including diarrhea.
[0007] Insensitivity to insulin exacerbates diabetes and obesity.
Insulin sensitivity is decreased by the interaction of adenosine
with A.sub.2B receptors. Thus, blocking the adenosine A.sub.2B
receptors of individuals with diabetes or obesity would benefit
patients with these disorders. It has also been demonstrated that
A.sub.2B-antagonists cause a reduction of blood glucose levels, and
thus would be particularly useful in the treatment of type-II
diabetes.
[0008] Another adverse biological effect of adenosine acting at the
A.sub.2B receptor is the over-stimulation of cerebral IL-6, a
cytokine associated with dementias and Alzheimer's disease.
Inhibiting the binding of adenosine to A.sub.2B receptors would
therefore mitigate those neurological disorders that are produced
by IL-6.
[0009] Type I hypersensitivity disorders, such as asthma, hay
fever, and atopic eczema, are stimulated by binding to
A.sub.2B-receptors of mast cells. Therefore, blocking these
adenosine receptors would provide a therapeutic benefit against
such disorders.
[0010] There are several compounds presently used in the treatment
of asthma. For example, theophylline is an effective antiasthmatic
agent, even though it is a poor adenosine receptor antagonist.
However, considerable plasma levels are needed for it to be
effective. Additionally, theophylline has substantial side effects,
most of which are due to its CNS action, which provide no
beneficial effects in asthma, and to the fact that it
non-specifically blocks all adenosine receptor subtypes.
[0011] Additionally adenosine treatment, such as inhaled adenosine
(or adenosine monophosphate), provokes bronchoconstriction in
asthmatics, but not in the normal population. This process is known
to involve mast cell activation, in that it releases mast cell
mediators, including histamine, PGD2-.beta.-hexosaminidase and
tryptase, and because it can be blocked by specific histamine
H.sub.1 blockers and chromolyn sodium. Accordingly, there is an
intrinsic difference in the way adenosine interacts with mast cells
from asthmatics, and thus A.sub.2B antagonists are particularly
useful in modulating mast cell function or in the activation of
human lung cells.
[0012] Accordingly, it is desired to provide compounds that are
potent A.sub.2B antagonists (i.e., compounds that inhibit the
A.sub.2B adenosine receptor), fully or partially selective for the
A.sub.2B receptor, useful in the treatment of various disease
states related to modulation of the A.sub.2B receptor, for example
cancer, asthma and diarrhea.
SUMMARY OF THE INVENTION
[0013] U.S. Non-Provisional patent application Ser. No. 10/431,167
discloses novel A.sub.2B adenosine receptor antagonists
[0014] A category of preferred compounds that fall within the scope
of this invention has been identified. Preferred compounds of
Formula I include those in which R.sup.1 and R.sup.2 are
independently optionally substituted lower alkyl, especially those
compounds in which R.sup.1 and R.sup.2 are different, and are lower
alkyl optionally substituted by cycloalkyl. More preferred are
those compounds in which X is pyrazol-4-yl, Y is methylene, and Z
is optionally substituted phenyl, especially phenyl substituted
with trifluoromethyl. Even more preferred are those compounds in
which R.sup.1 and R.sup.2 are chosen from ethyl, n-propyl
cyclopropylmethyl, or iso-butyl, especially those in which R.sup.1
is n-propyl and R.sup.2 is ethyl. A preferred Z is
3-trifluoromethylphenyl. Consequently, novel processes for the
preparation of such compounds have been developed.
[0015] Accordingly, in a first aspect, the invention relates to a
process for the preparation of a compound of the formula:
##STR00001##
wherein: [0016] R.sup.1 and R.sup.2 are independently optionally
substituted alkyl; [0017] X is optionally substituted arylene or
optionally substituted heteroarylene; [0018] Y is a covalent bond
or lower alkylene; and [0019] Z is optionally substituted
monocyclic aryl or optionally substituted monocyclic heteroaryl;
comprising; contacting a compound of the formula:
##STR00002##
[0019] in which R.sup.1, R.sup.2, X, Y, and Z are as defined above;
with a base.
[0020] In a preferred embodiment, the compound of formula (3) is
contacted with a base, preferably in a protic solvent. The base is
preferably chosen from sodium hydroxide, potassium hydroxide,
sodium methoxide, sodium ethoxide, and potassium t-butoxide
preferably aqueous sodium hydroxide solution, and the protic
solvent is preferably methanol.
[0021] In a second aspect, the invention relates to a process for
the preparation of a compound of formula (3):
##STR00003##
in which R.sup.1, R.sup.2, X, Y and Z are as defined above;
comprising: contacting a compound of the formula (2);
##STR00004##
with a carboxylic acid of the formula Z-Y--X--CO.sub.2H; in which
X, Y and Z are as defined above.
[0022] In a preferred embodiment, the compound of formula (2) is
contacted with a compound of the formula Z-Y--X--CO.sub.2H in a
polar solvent, preferably methanol, in the presence of a coupling
agent used to form amide bonds, preferably a carbodiimide
derivative. In a more preferred embodiment the carbodiimide
derivative is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide.
Alternatively, the compound of formula (2) is contacted with an
acid halide of the formula Z-Y--X--C(O)L, where L is chloro or
bromo.
[0023] In a third aspect, the invention relates to a process for
the preparation of a compound of formula (3):
##STR00005##
in which R.sup.1, R.sup.2, X, Y and Z are as defined above;
comprising: contacting a compound of the formula (16);
##STR00006##
in which R.sup.2, X, Y and Z are as defined above; with a compound
of the formula R.sup.1L, in which L is a leaving group.
[0024] In a preferred embodiment, the compound of formula (16) is
contacted with an alkyl halide, preferably an alkyl iodide, in the
presence of a base, preferably potassium carbonate, in a polar
solvent, preferably N,N-dimethylformamide.
[0025] In a fourth aspect, the invention relates to a process for
the preparation of a compound of formula (3):
##STR00007##
in which R.sup.1, R.sup.2, X, Y and Z are as defined above;
comprising: contacting a compound of formula (13)
##STR00008##
in which R.sup.1, X, Y and Z are as defined above; with a compound
of the formula R.sup.2L, in which L is a leaving group.
[0026] In a preferred embodiment, the compound of formula (13) is
contacted with an alkyl halide, preferably an alkyl iodide, in the
presence of a base, preferably potassium carbonate, in a polar
solvent, preferably N,N-dimethylformamide.
[0027] In a fifth aspect, the invention relates to a process for
the preparation of a compound of formula (16):
##STR00009##
in which R.sup.2, X, Y and Z are as defined above; comprising:
contacting a compound of formula (15):
##STR00010##
in which R.sup.2 is as defined above; with a carboxylic acid of the
formula Z-Y--X--CO.sub.2H; in which X, Y and Z are as defined
above.
[0028] In a preferred embodiment, the compound of formula (15) is
contacted with a compound of the formula Z-Y--X--CO.sub.2H in a
polar solvent, preferably methanol, in the presence of a coupling
agent used to form amide bonds, preferably a carbodiimide
derivative. In a more preferred embodiment the carbodiimide
derivative is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide.
Alternatively, the compound of formula (15) is contacted with an
acid halide of the formula Z-Y--X--C(O)L, where L is chloro or
bromo.
[0029] In a sixth aspect, the invention relates to a process for
the preparation of a compound of formula (13):
##STR00011##
in which R.sup.1, X, Y and Z are as defined above; comprising:
contacting a compound of formula (12)
##STR00012##
in which R.sup.1 is as defined above; with a carboxylic acid of the
formula Z-Y--X--CO.sub.2H; in which X, Y and Z are as defined
above.
[0030] In a preferred embodiment, the compound of formula (12) is
contacted with a compound of the formula Z-Y--X--CO.sub.2H in a
polar solvent, preferably methanol, in the presence of a coupling
agent used to form amide bonds, preferably a carbodiimide
derivative. In a more preferred embodiment the carbodiimide
derivative is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide.
Alternatively, the compound of formula (12) is contacted with an
acid halide of the formula Z-Y--X--C(O)L, where L is chloro or
bromo.
[0031] In a seventh aspect, the invention relates to a process for
the preparation of a compound of formula (15):
##STR00013##
in which R.sup.2 is as defined above; comprising the steps of: 1)
contacting a compound of formula (4):
##STR00014##
with ethyl cyanoacetate in the presence of a base in a protic
solvent, preferably ethanol/sodium ethoxide; 2) contacting the
product thus formed:
##STR00015##
with a mixture of sodium nitrite in acetic acid/water; and 3)
contacting the product thus formed:
##STR00016##
with a mixture of aqueous ammonia and sodium dithionite.
[0032] In an eighth aspect, the invention relates to a process for
the preparation of a compound of formula (12):
##STR00017##
in which R.sup.1 is as defined above; comprising the steps of: 1)
contacting a compound of the formula:
##STR00018##
with a) hexamethyldisilazane followed by b) R.sup.1L, where R.sup.1
is as defined above and L is a leaving group; 2) contacting the
compound thus formed:
##STR00019##
with a mixture of sodium nitrite in acetic acid/water; and 3)
contacting the product thus formed.
##STR00020##
with a mixture of aqueous ammonia and sodium dithionite.
[0033] In a ninth aspect, the invention relates to a process for
the preparation of a compound of formula (2):
##STR00021##
in which R.sup.1 and R.sup.2 are as defined above; comprising the
steps of: 1) contacting a compound of the formula:
##STR00022##
with ethyl cyanoacetate in the presence of a base in a protic
solvent, preferably ethanol/sodium ethoxide; 2) contacting the
product thus formed:
##STR00023##
with the dimethylacetal of N,N-dimethylformamide; 3) contacting the
product thus formed:
##STR00024##
with a compound of formula R.sup.1L, in which L is a leaving group,
preferably an iodide, in the presence of a base, preferably
potassium carbonate, in a polar solvent, preferably
N,N-dimethylformamide. 4) contacting the product thus formed:
##STR00025##
with aqueous ammonia; 5) contacting the product thus formed:
##STR00026##
with a mixture of sodium nitrite in acetic acid/water; and 6)
contacting the product thus formed:
##STR00027##
with a mixture of aqueous ammonia and sodium dithionite.
[0034] In a tenth aspect, the invention relates to a process for
the preparation of a compound of formula (7)
##STR00028##
comprising the steps of: 1) contacting a compound of the
formula:
##STR00029##
with the dimethylacetal of N,N-dimethylformamide; 2) contacting the
product thus formed:
##STR00030##
with a compound of formula R.sup.2L, in which L is a leaving group,
preferably an iodide, in the presence of a base, preferably
potassium carbonate, in a polar solvent, preferably
N,N-dimethylformamide; and 3) contacting the product thus
formed:
##STR00031##
with aqueous ammonia.
[0035] In an eleventh aspect, the invention relates to a novel
intermediate of the formula:
##STR00032##
in which R.sup.1 is n-propyl, 2-methylpropyl, or cyclopropylmethyl
and R.sup.2 is methyl or ethyl. Particularly preferred is the
compound in which R.sup.1 is n-propyl and R.sup.2 is ethyl:
##STR00033##
namely
6-[2-(dimethylamino)-1-azavinyl]-1-ethyl-3-propyl-1,3-dihydropyrim-
idine-2,4-dione.
[0036] In a twelfth aspect, the invention relates to a novel
intermediate of the formula:
##STR00034##
in which R.sup.1 is n-propyl or cyclopropylmethyl, preferably
n-propyl
##STR00035##
namely
N-(6-amino-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[3-(t-
rifluoromethyl)-phenyl]methyl}pyrazol-4-yl)carboxamide.
[0037] In a thirteenth aspect, the invention relates to a novel
intermediate of the formula:
##STR00036##
in which R.sup.2 is methyl or ethyl, preferably ethyl;
##STR00037##
namely
N-(6-amino-1-ethyl-2,4-dioxo(1,3-dihydropyrimidin-5-yl))(1-{[3-(tr-
ifluoromethyl)phenyl]methyl}pyrazol-4-yl)carboxamide;
[0038] In a fourteenth aspect, the invention relates to a novel
intermediate of the formula:
##STR00038##
in which R.sup.1 is n-propyl, 2-methylpropyl, or cyclopropylmethyl
and R.sup.2 is methyl or ethyl. Particularly preferred is the
intermediate in which R.sup.1 is n-propyl and R.sup.2 is ethyl;
##STR00039##
namely
N-(6-amino-1-ethyl-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(-
1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)carboxamide.
[0039] Other aspects of the invention relates to pharmaceutical
formulations, comprising a therapeutically effective amount of a
compound of Formula I and at least one pharmaceutically acceptable
excipient.
[0040] A further aspect of this invention relates to a method of
using the compounds of Formula I in the treatment of a disease or
condition in a mammal that is treatable by inhibiting an adenosine
receptor characterized as A.sub.2B, comprising administering to a
mammal in need thereof a therapeutically effective dose of a
compound of Formula I. Such diseases include, but are not limited
to, at least one of asthma, inflammatory gastrointestinal tract
disorders, including diarrhea, cardiovascular diseases such as
atherosclerosis, neurological disorders such as senile dementia,
Alzheimer's disease, and Parkinson's disease, and diseases related
to angiogenesis, for example diabetic retinopathy and cancer.
[0041] One preferred group of compounds of Formula I are those in
which R.sup.1 and R.sup.2 are different and are independently lower
alkyl optionally substituted by cycloalkyl. Within this group, a
first preferred class of compounds include those in which R.sup.1
is lower alkyl of 2-4 carbon atoms optionally substituted by
cyclopropyl and R.sup.2 is lower alkyl of 2-4 carbon atoms,
particularly where R.sup.1 and R.sup.2 are chosen from ethyl and
n-propyl, and X is optionally substituted pyrazolen-1,4-yl. Within
this class, a preferred subclass of compounds is where Y is lower
alkylene, preferably methylene, and Z is optionally substituted
phenyl, preferably 3-trifluoromethylphenyl. Most preferred are
those compounds of Formula I in which R.sup.1 is n-propyl and
R.sup.2 is ethyl.
[0042] At present, the preferred compounds are: [0043]
3-ethyl-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,-
3,7-trihydropurine-2,6-dione; [0044]
1-cyclopropylmethyl-3-methyl-8-[1-(phenylmethyl)pyrazol-4-yl]-1,3,7-trihy-
dropurine-2,6-dione; [0045]
1-cyclopropylmethyl-3-methyl-8-{1-[(3-trifluoromethylphenyl)methyl]pyrazo-
l-4-yl)-1,3,7-trihydropurine-2,6-dione; [0046]
1-cyclopropylmethyl-3-ethyl-8-(1-[(3-trifluoromethylphenyl)methyl]pyrazol-
-4-yl}-1,3,7-trihydropurine-2,6-dione; [0047]
1-cyclopropylmethyl-3-methyl-8-{1-[(3-fluorophenyl)methyl]pyrazol-4-yl}-1-
,3,7-trihydropurine-2,6-dione; [0048]
1-cyclopropylmethyl-3-ethyl-8-{1-[(3-fluorophenyl)methyl]pyrazol-4-yl}-1,-
3,7-trihydropurine-2,6-dione; [0049]
1-cyclopropylmethyl-3-ethyl-8-(1-{[6-(trifluoromethyl)(3-pyridyl)]methyl}-
pyrazol-4-yl)-1,3,7-trihydropurine-2,6-dione; [0050]
3-({4-[1-(cyclopropylmethyl)-3-methyl-2,6-dioxo-1,3,7-trihydropurin-8-yl]-
pyrazolyl}methyl)benzenecarbonitrile; [0051]
8-[1-(2-(1H-1,2,3,4-tetraazol-5-yl)ethyl)pyrazol-4-yl]-3-methyl-1-cyclopr-
opylmethyl-1,3,7-trihydropurine-2,6-dione; [0052]
1-(2-methylpropyl)-3-methyl-8-[1-benzylpyrazol-4-yl]-1,3,7-trihydropurine-
-2,6-dione; [0053]
1-(2-methylpropyl)-3-ethyl-8-(1-[(3-fluorophenyl)methyl]pyrazol-4-yl)-1,3-
,7-trihydropurine-2,6-dione; [0054]
1-(2-methylpropyl)-3-methyl-8-(1-[(3-trifluoromethylphenyl)methyl]pyrazol-
-4-yl}-1,3,7-trihydropurine-2,6-dione; [0055]
1-(2-methylpropyl)-3-methyl-8-(1-[(3-fluorophenyl)methyl]pyrazol-4-yl)-1,-
3,7-trihydropurine-2,6-dione; [0056]
3-ethyl-1-(2-methylpropyl)-8-(1-{[6-(trifluoromethyl)(3-pyridyl)]methyl}p-
yrazol-4-yl)-1,3,7-trihydropurine-2,6-dione; [0057]
1-ethyl-3-methyl-8-{1-[(3-fluorophenyl)methyl]pyrazol-4-yl}-1,3,7-trihydr-
opurine-2,6-dione; and [0058]
3-ethyl-1-propyl-8-[1-(2-pyridylmethyl)pyrazol-4-yl]-1,3,7-trihydropurine-
-2,6-dione.
[0059] Particularly preferred is
3-ethyl-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]-methyl}pyrazol-4-yl)-1-
,3,7-trihydropurine-2,6-dione.
DEFINITIONS AND GENERAL PARAMETERS
[0060] As used in the present specification, the following words
and phrases are generally intended to have the meanings as set
forth below, except to the extent that the context in which they
are used indicates otherwise.
[0061] The term "alkyl" refers to a monoradical branched or
unbranched saturated hydrocarbon chain having 1, 2, 3, 4, 5, 6, 7,
8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 carbon atoms.
This term is exemplified by groups such as methyl, ethyl, n-propyl,
iso-propyl, n-butyl, iso-butyl, t-butyl, n-hexyl, n-decyl,
tetradecyl, and the like.
[0062] The term "substituted alkyl" refers to: [0063] 1) an alkyl
group as defined above, having 1, 2, 3, 4 or 5 substituents,
preferably 1 to 3 substituents, selected from the group consisting
of alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl,
acylamino, acyloxy, amino, aminocarbonyl, alkoxycarbonylamino,
azido, cyano, halogen, hydroxy, keto, thiocarbonyl, carboxy,
carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio, thiol,
alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl,
aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy,
hydroxyamino, alkoxyamino, nitro, --SO-alkyl, --SO-aryl,
--SO-heteroaryl, --SO.sub.2-alkyl, SO.sub.2-aryl and
--SO.sub.2-heteroaryl. Unless otherwise constrained by the
definition, all substituents may optionally be further substituted
by 1, 2, or 3 substituents chosen from alkyl, carboxy,
carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF.sub.3,
amino, substituted amino, cyano, and --S(O).sub.nR, where R is
alkyl, aryl, or heteroaryl and n is 0, 1 or 2; or [0064] 2) an
alkyl group as defined above that is interrupted by 1-10 atoms
independently chosen from oxygen, sulfur and NR.sub.a--, where
R.sub.a is chosen from hydrogen, alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclyl. All
substituents may be optionally further substituted by alkyl,
alkoxy, halogen, CF.sub.3, amino, substituted amino, cyano, or
S(O).sub.nR, in which R is alkyl, aryl, or heteroaryl and n is 0, 1
or 2; or [0065] 3) an alkyl group as defined above that has both 1,
2, 3, 4 or 5 substituents as defined above and is also interrupted
by 1-10 atoms as defined above.
[0066] The term "lower alkyl" refers to a monoradical branched or
unbranched saturated hydrocarbon chain having 1, 2, 3, 4, 5, or 6
carbon atoms. This term is exemplified by groups such as methyl,
ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, t-butyl, n-hexyl,
and the like.
[0067] The term "substituted lower alkyl" refers to lower alkyl as
defined above having 1 to 5 substituents, preferably 1, 2, or 3
substituents, as defined for substituted alkyl, or a lower alkyl
group as defined above that is interrupted by 1, 2, 3, 4, or 5
atoms as defined for substituted alkyl, or a lower alkyl group as
defined above that has both 1, 2, 3, 4 or 5 substituents as defined
above and is also interrupted by 1, 2, 3, 4, or 5 atoms as defined
above.
[0068] The term "alkylene" refers to a diradical of a branched or
unbranched saturated hydrocarbon chain, having 1, 2, 3, 4, 5, 6, 7,
8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 carbon atoms,
preferably 1-10 carbon atoms, more preferably 1, 2, 3, 4, 5 or 6
carbon atoms. This term is exemplified by groups such as methylene
(--CH.sub.2--), ethylene (--CH.sub.2CH.sub.2--), the propylene
isomers (e.g., --CH.sub.2CH.sub.2CH.sub.2-- and
--CH(CH.sub.3)CH.sub.2--) and the like.
[0069] The term "lower alkylene" refers to a diradical of a
branched or unbranched saturated hydrocarbon chain, preferably
having from 1, 2, 3, 4, 5, or 6 carbon atoms.
[0070] The term "lower alkylene" refers to a diradical of a
branched or unbranched saturated hydrocarbon chain, preferably
having from 1, 2, 3, 4, 5, or 6 carbon atoms.
[0071] The term "substituted alkylene" refers to: [0072] (1) an
alkylene group as defined above having 1, 2, 3, 4, or 5
substituents selected from the group consisting of alkyl, alkenyl,
alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino,
acyloxy, amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano,
halogen, hydroxy, keto, thiocarbonyl, carboxy, carboxyalkyl,
arylthio, heteroarylthio, heterocyclylthio, thiol, alkylthio, aryl,
aryloxy, heteroaryl, aminosulfonyl, aminocarbonylamino,
heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino,
alkoxyamino, nitro, --SO-alkyl, --SO-aryl, --SO-heteroaryl,
--SO.sub.2-alkyl, SO.sub.2-aryl and --SO.sub.2-heteroaryl. Unless
otherwise constrained by the definition, all substituents may
optionally be further substituted by 1, 2, or 3 substituents chosen
from alkyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy,
halogen, CF.sub.3, amino, substituted amino, cyano, and
S(O).sub.nR, where R is alkyl, aryl, or heteroaryl and n is 0, 1 or
2; or [0073] (2) an alkylene group as defined above that is
interrupted by 1-20 atoms independently chosen from oxygen, sulfur
and NR.sub.a--, where R.sub.a is chosen from hydrogen, optionally
substituted alkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl and
heterocycyl, or groups selected from carbonyl, carboxyester,
carboxyamide and sulfonyl; or [0074] (3) an alkylene group as
defined above that has both 1, 2, 3, 4 or 5 substituents as defined
above and is also interrupted by 1-20 atoms as defined above.
Examples of substituted alkylenes are chloromethylene (--CH(Cl)--),
aminoethylene (--CH(NH.sub.2)CH.sub.2--), methylaminoethylene
(--CH(NHMe)CH.sub.2--), 2-carboxypropylene isomers
(--CH.sub.2CH(CO.sub.2H)CH.sub.2--), ethoxyethyl
(--CH.sub.2CH.sub.2O--CH.sub.2CH.sub.2--), ethylmethylaminoethyl
(--CH.sub.2CH.sub.2N(CH.sub.3)CH.sub.2CH.sub.2--),1-ethoxy-2-(2-ethoxy-et-
hoxy)ethane
(--CH.sub.2CH.sub.2O--CH.sub.2CH.sub.2--OCH.sub.2CH.sub.2--OCH.sub.2CH.su-
b.2--), and the like.
[0075] The term "aralkyl" refers to an aryl group covalently linked
to an alkylene group, where aryl and alkylene are defined herein.
"Optionally substituted aralkyl" refers to an optionally
substituted aryl group covalently linked to an optionally
substituted alkylene group. Such aralkyl groups are exemplified by
benzyl, phenylethyl, 3-(4-methoxyphenyl)propyl, and the like.
[0076] The term "alkoxy" refers to the group R--O--, where R is
optionally substituted alkyl or optionally substituted cycloalkyl,
or R is a group --Y-Z, in which Y is optionally substituted
alkylene and Z is optionally substituted alkenyl, optionally
substituted alkynyl; or optionally substituted cycloalkenyl, where
alkyl, alkenyl, alkynyl, cycloalkyl and cycloalkenyl are as defined
herein. Preferred alkoxy groups are optionally substituted
alkyl-O-- and include, by way of example, methoxy, ethoxy,
n-propoxy, iso-propoxy, n-butoxy, tert-butoxy, sec-butoxy (or
isobutoxy), n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy,
trifluoromethoxy, and the like.
[0077] The term "alkylthio" refers to the group R--S--, where R is
as defined for alkoxy.
[0078] The term "alkenyl" refers to a monoradical of a branched or
unbranched unsaturated hydrocarbon group preferably having from 2
to 20 carbon atoms, more preferably 2 to 10 carbon atoms and even
more preferably 2 to 6 carbon atoms and having 1-6, preferably 1,
double bond (vinyl). Preferred alkenyl groups include ethenyl or
vinyl (--CH.dbd.CH.sub.2), 1-propylene or allyl
(--CH.sub.2CH.dbd.CH.sub.2), isopropylene
(--C(CH.sub.3).dbd.CH.sub.2), bicyclo[2.2.1]heptene, and the like.
In the event that alkenyl is attached to nitrogen, the double bond
cannot be alpha to the nitrogen.
[0079] The term "lower alkenyl" refers to alkenyl as defined above
having from 2 to 6 carbon atoms.
[0080] The term "substituted alkenyl" refers to an alkenyl group as
defined above having 1, 2, 3, 4 or 5 substituents, and preferably
1, 2, or 3 substituents, selected from the group consisting of
alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl,
acylamino, acyloxy, amino, aminocarbonyl, alkoxycarbonylamino,
azido, cyano, halogen, hydroxy, keto, thiocarbonyl, carboxy,
carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio, thiol,
alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl,
aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy,
hydroxyamino, alkoxyamino, nitro, --SO-alkyl, --SO-aryl,
--SO-heteroaryl, --SO.sub.2-alkyl, SO.sub.2-aryl and
--SO.sub.2-heteroaryl. Unless otherwise constrained by the
definition, all substituents may optionally be further substituted
by 1, 2, or 3 substituents chosen from alkyl, carboxy,
carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF.sub.3,
amino, substituted amino, cyano, and --S(O).sub.nR, where R is
alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
[0081] The term "alkynyl" refers to a monoradical of an unsaturated
hydrocarbon, preferably having from 2 to 20 carbon atoms, more
preferably 2 to 10 carbon atoms and even more preferably 2 to 6
carbon atoms and having at least 1 and preferably from 1-6 sites of
acetylene (triple bond) unsaturation. Preferred alkynyl groups
include ethynyl, (--C.ident.CH), propargyl (or prop-1-yn-3-yl,
--CH.sub.2C.ident.CH), and the like. In the event that alkynyl is
attached to nitrogen, the triple bond cannot be alpha to the
nitrogen.
[0082] The term "substituted alkynyl" refers to an alkynyl group as
defined above having 1, 2, 3, 4 or 5 substituents, and preferably
1, 2, or 3 substituents, selected from the group consisting of
alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl,
acylamino, acyloxy, amino, aminocarbonyl, alkoxycarbonylamino,
azido, cyano, halogen, hydroxy, keto, thiocarbonyl, carboxy,
carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio, thiol,
alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl,
aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy,
hydroxyamino, alkoxyamino, nitro, --SO-alkyl, --SO-aryl,
--SO-heteroaryl, --SO.sub.2-alkyl, SO.sub.2-aryl and
--SO.sub.2-heteroaryl. Unless otherwise constrained by the
definition, all substituents may optionally be further substituted
by 1, 2, or 3 substituents chosen from alkyl, carboxy;
carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF.sub.3,
amino, substituted amino, cyano, and --S(O).sub.nR, where R is
alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
[0083] The term "aminocarbonyl" refers to the group --C(O)NRR where
each R is independently hydrogen, alkyl, aryl, heteroaryl,
heterocyclyl or where both R groups are joined to form a
heterocyclic group (e.g., morpholino). Unless otherwise constrained
by the definition, all substituents may optionally be further
substituted by 1-3 substituents chosen from alkyl, carboxy,
carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF.sub.3,
amino, substituted amino, cyano, and --S(O).sub.nR, where R is
alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
[0084] The term "acylamino" refers to the group --NRC(O)R where
each R is independently hydrogen, alkyl, aryl, heteroaryl, or
heterocyclyl. Unless otherwise constrained by the definition, all
substituents may optionally be further substituted by 1-3
substituents chosen from alkyl, carboxy, carboxyalkyl,
aminocarbonyl, hydroxy, alkoxy, halogen, CF.sub.3, amino,
substituted amino, cyano, and --S(O).sub.nR, where R is alkyl,
aryl, or heteroaryl and n is 0, 1 or 2.
[0085] The term "acyloxy" refers to the groups --O(O)C-alkyl,
--O(O)C-cycloalkyl, --O(O)C-aryl, --O(O)C-heteroaryl, and
--O(O)C-heterocyclyl. Unless otherwise constrained by the
definition, all substituents may be optionally further substituted
by alkyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy,
halogen, CF.sub.3, amino, substituted amino, cyano, or
--S(O).sub.nR, where R is alkyl, aryl, or heteroaryl and n is 0, 1
or 2.
[0086] The term "aryl" refers to an aromatic carbocyclic group of 6
to 20 carbon atoms having a single ring (e.g., phenyl) or multiple
rings (e.g., biphenyl), or multiple condensed (fused) rings (e.g.,
naphthyl or anthryl). Preferred aryls include phenyl, naphthyl and
the like.
[0087] The term "arylene" refers to a diradical of an aryl group as
defined above. This term is exemplified by groups such as
1,4-phenylene, 1,3-phenylene, 1,2-phenylene, 1,4'-biphenylene, and
the like.
[0088] Unless otherwise constrained by the definition for the aryl
or arylene substituent, such aryl or arylene groups can optionally
be substituted with from 1 to 5 substituents, preferably 1 to 3
substituents, selected from the group consisting of alkyl, alkenyl,
alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino,
acyloxy, amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano,
halogen, hydroxy, keto, thiocarbonyl, carboxy, carboxyalkyl,
arylthio, heteroarylthio, heterocyclylthio, thiol, alkylthio, aryl,
aryloxy, heteroaryl, aminosulfonyl, aminocarbonylamino,
heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino,
alkoxyamino, nitro, --SO-alkyl, --SO-aryl, --SO-heteroaryl,
--SO.sub.2-alkyl, SO.sub.2-aryl and --SO.sub.2-heteroaryl. Unless
otherwise constrained by the definition, all substituents may
optionally be further substituted by 1-3 substituents chosen from
alkyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy,
halogen, CF.sub.3, amino, substituted amino, cyano, and
--S(O).sub.nR, where R is alkyl, aryl, or heteroaryl and n is 0, 1
or 2.
[0089] The term "aryloxy" refers to the group aryl-O-- wherein the
aryl group is as defined above, and includes optionally substituted
aryl groups as also defined above. The term "arylthio" refers to
the group R--S--, where R is as defined for aryl.
[0090] The term "amino" refers to the group --NH.sub.2.
[0091] The term "substituted amino" refers to the group --NRR where
each R is independently selected from the group consisting of
hydrogen, alkyl, cycloalkyl, carboxyalkyl (for example,
benzyloxycarbonyl), aryl, heteroaryl and heterocyclyl provided that
both R groups are not hydrogen, or a group --Y-Z, in which Y is
optionally substituted alkylene and Z is alkenyl, cycloalkenyl, or
alkynyl, Unless otherwise constrained by the definition, all
substituents may optionally be further substituted by 1-3
substituents chosen from alkyl, carboxy, carboxyalkyl,
aminocarbonyl, hydroxy, alkoxy, halogen, CF.sub.3, amino,
substituted amino, cyano, and --S(O).sub.nR, where R is alkyl,
aryl, or heteroaryl and n is 0, 1 or 2.
[0092] The term "carboxyalkyl" refers to the groups --C(O)O-alkyl
or --C(O)O-cycloalkyl, where alkyl and cycloalkyl, are as defined
herein, and may be optionally further substituted by alkyl,
alkenyl, alkynyl, alkoxy, halogen, CF.sub.3, amino, substituted
amino, cyano, or --S(O).sub.nR, in which R is alkyl, aryl, or
heteroaryl and n is 0, 1 or 2.
[0093] The term "cycloalkyl" refers to carbocyclic groups of from 3
to 20 carbon atoms having a single cyclic ring or multiple
condensed rings. Such cycloalkyl groups include, by way of example,
single ring structures such as cyclopropyl, cyclobutyl,
cyclopentyl, cyclooctyl, and the like, or multiple ring structures
such as adamantanyl, bicyclo[2.2.1]heptane,
1,3,3-trimethylbicyclo[2.2.1]hept-2-yl,
(2,3,3-trimethylbicyclo[2.2.1]hept-2-yl), or carbocyclic groups to
which is fused an aryl group, for example indane, and the like.
[0094] The term "substituted cycloalkyl" refers to cycloalkyl
groups having 1, 2, 3, 4 or 5 substituents, and preferably 1, 2, or
3 substituents, selected from the group consisting of alkyl,
alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl,
acylamino, acyloxy, amino, aminocarbonyl, alkoxycarbonylamino,
azido, cyano, halogen, hydroxy, keto, thiocarbonyl, carboxy,
carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio, thiol,
alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl,
aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy,
hydroxyamino, alkoxyamino, nitro, --SO-alkyl, --SO-aryl,
--SO-heteroaryl, --SO.sub.2-alkyl, SO.sub.2-aryl and
--SO.sub.2-heteroaryl. Unless otherwise constrained by the
definition, all substituents may optionally be further substituted
by 1, 2, or 3 substituents chosen from alkyl, carboxy,
carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF.sub.3,
amino, substituted amino, cyano, and --S(O).sub.nR, where R is
alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
[0095] The term "halogen" or "halo" refers to fluoro, bromo,
chloro, and iodo.
[0096] The term "acyl" denotes a group --C(O)R, in which R is
hydrogen, optionally substituted alkyl, optionally substituted
cycloalkyl, optionally substituted heterocyclyl, optionally
substituted aryl, and optionally substituted heteroaryl.
[0097] The term "heteroaryl" refers to a radical derived from an
aromatic cyclic group (i.e., fully unsaturated) having 1, 2, 3, 4,
5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 carbon atoms and 1, 2, 3
or 4 heteroatoms selected from oxygen, nitrogen and sulfur within
at least one ring. Such heteroaryl groups can have a single ring
(e.g., pyridyl or furyl) or multiple condensed rings (e.g.,
indolizinyl, benzothiazolyl, or benzothienyl). Examples of
heteroaryls include, but are not limited to, [1,2,4]oxadiazole,
[1,3,4]oxadiazole, [1,2,4]thiadiazole, [1,3,4]thiadiazole, pyrrole,
imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine,
indolizine, isoindole, indole, indazole, purine, quinolizine,
isoquinoline, quinoline, phthalazine, naphthylpyridine,
quinoxaline, quinazoline, cinnoline, pteridine, carbazole,
carboline, phenanthridine, acridine, phenanthroline, isothiazole,
phenazine, isoxazole, phenoxazine, phenothiazine, imidazolidine,
imidazoline, and the like as well as N-oxide and N-alkoxy
derivatives of nitrogen containing heteroaryl compounds, for
example pyridine-N-oxide derivatives.
[0098] The term "heteroarylene" refers to a diradical of a
heteroaryl group as defined above. This term is exemplified by
groups such as 2,5-imidazolene, 3,5-[1,2,4]oxadiazolene,
2,4-oxazolene, 1,4-pyrazolene, and the like. For example,
1,4-pyrazolene is:
##STR00040##
where A represents the point of attachment.
[0099] Unless otherwise constrained by the definition for the
heteroaryl or heteroarylene substituent, such heteroaryl or
heterarylene groups can be optionally substituted with 1 to 5
substituents, preferably 1 to 3 substituents selected from the
group consisting of alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl,
cycloalkenyl, acyl, acylamino, acyloxy, amino, aminocarbonyl,
alkoxycarbonylamino, azido, cyano, halogen, hydroxy, keto,
thiocarbonyl, carboxy, carboxyalkyl, arylthio, heteroarylthio,
heterocyclylthio, thiol, alkylthio, aryl, aryloxy, heteroaryl,
aminosulfonyl, aminocarbonylamino, heteroaryloxy, heterocyclyl,
heterocyclooxy, hydroxyamino, alkoxyamino, nitro, --SO-alkyl,
--SO-aryl, --SO-heteroaryl, --SO.sub.2-alkyl, SO.sub.2-aryl and
--SO.sub.2-heteroaryl. Unless otherwise constrained by the
definition, all substituents may optionally be further substituted
by 1-3 substituents chosen from alkyl, carboxy, carboxyalkyl,
aminocarbonyl, hydroxy, alkoxy, halogen, CF.sub.3, amino,
substituted amino, cyano, and --S(O).sub.nR, where R is alkyl,
aryl, or heteroaryl and n is 0, 1 or 2.
[0100] The term "heteroaralkyl" refers to a heteroaryl group
covalently linked to an alkylene group, where heteroaryl and
alkylene are defined herein. "Optionally substituted heteroaralkyl"
refers to an optionally substituted heteroaryl group covalently
linked to an optionally substituted alkylene group. Such
heteroaralkyl groups are exemplified by 3-pyridylmethyl,
quinolin-8-ylethyl, 4-methoxythiazol-2-ylpropyl, and the like.
[0101] The term "heteroaryloxy" refers to the group
heteroaryl-O--.
[0102] The term "heterocyclyl" refers to a monoradical saturated or
partially unsaturated group having a single ring or multiple
condensed rings, having from 1 to 40 carbon atoms and from 1 to 10
hetero atoms, preferably 1, 2, 3 or 4 hetero atoms, selected from
nitrogen, sulfur, phosphorus, and/or oxygen within the ring.
Heterocyclic groups can have a single ring or multiple condensed
rings, and include tetrahydrofuranyl, morpholino, piperidinyl,
piperazino, dihydropyridino, and the like.
[0103] Unless otherwise constrained by the definition for the
heterocyclic substituent, such heterocyclic groups can be
optionally substituted with 1, 2, 3, 4 or 5, and preferably 1, 2 or
3 substituents, selected from the group consisting of alkyl,
alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl,
acylamino, acyloxy, amino, aminocarbonyl, alkoxycarbonylamino,
azido, cyano, halogen, hydroxy, keto, thiocarbonyl, carboxy,
carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio, thiol,
alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl,
aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy,
hydroxyamino, alkoxyamino, nitro, --SO-alkyl, --SO-aryl,
--SO-heteroaryl, SO.sub.2-alkyl, SO.sub.2-aryl and
--SO.sub.2-heteroaryl. Unless otherwise constrained by the
definition, all substituents may optionally be further substituted
by 1-3 substituents chosen from alkyl, carboxy, carboxyalkyl,
aminocarbonyl, hydroxy, alkoxy, halogen, CF.sub.3, amino,
substituted amino, cyano, and --S(O).sub.nR, where R is alkyl,
aryl, or heteroaryl and n is 0, 1 or 2.
[0104] The term "thiol" refers to the group --SH.
[0105] The term "substituted alkylthio" refers to the group
--S-substituted alkyl.
[0106] The term "heteroarylthiol" refers to the group
--S-heteroaryl wherein the heteroaryl group is as defined above
including optionally substituted heteroaryl groups as also defined
above.
[0107] The term "sulfoxide" refers to a group --S(O)R, in which R
is alkyl, aryl, or heteroaryl. "Substituted sulfoxide" refers to a
group --S(O)R, in which R is substituted alkyl, substituted aryl,
or substituted heteroaryl, as defined herein.
[0108] The term "sulfone" refers to a group --S(O).sub.2R, in which
R is alkyl, aryl, or heteroaryl. "Substituted sulfone" refers to a
group --S(O).sub.2R, in which R is substituted alkyl, substituted
aryl, or substituted heteroaryl, as defined herein.
[0109] The term "keto" refers to a group --C(O)--. The term
"thiocarbonyl" refers to a group --C(S)--. The term "carboxy"
refers to a group --C(O)--OH.
[0110] The term "coupling agent used to form amide bonds" refers to
those compounds that are conventionally employed to facilitate
formation of amide bonds through the reaction of a carboxylic acid
and an amine. Examples of such coupling agents are
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide,
1,3-di-t-butylcarbodiimide, 1,3-dicyclohexylcarbodiimide, and the
like.
[0111] The term "leaving group" is used in the conventional manner,
and refers to a moiety that is capable of being displaced by a
nucleophile in a replacement or substitution reaction. Examples of
leaving groups are chloro, bromo, iodo, mesylate, tosylate, and the
like.
[0112] "Optional" or "optionally" means that the subsequently
described event or circumstance may or may not occur, and that the
description includes instances where said event or circumstance
occurs and instances in which it does not.
[0113] The term "compound of Formula I" is intended to encompass
the compounds of the invention as disclosed, and the
pharmaceutically acceptable salts, pharmaceutically acceptable
esters, prodrugs, hydrates and polymorphs of such compounds.
Additionally, the compounds of the invention may possess one or
more asymmetric centers, and can be produced as a racemic mixture
or as individual enantiomers or diastereoisomers. The number of
stereoisomers present in any given compound of Formula I depends
upon the number of asymmetric centers present (there are 2.sup.n
stereoisomers possible where n is the number of asymmetric
centers). The individual stereoisomers may be obtained by resolving
a racemic or non-racemic mixture of an intermediate at some
appropriate stage of the synthesis, or by resolution of the
compound of Formula I by conventional means. The individual
stereoisomers (including individual enantiomers and
diastereoisomers) as well as racemic and non-racemic mixtures of
stereoisomers are encompassed within the scope of the present
invention, all of which are intended to be depicted by the
structures of this specification unless otherwise specifically
indicated.
[0114] "Isomers" are different compounds that have the same
molecular formula.
[0115] "Stereoisomers" are isomers that differ only in the way the
atoms are arranged in space.
[0116] "Enantiomers" are a pair of stereoisomers that are
non-superimposable mirror images of each other. A 1:1 mixture of a
pair of enantiomers is a "racemic" mixture. The term "(.+-.)" is
used to designate a racemic mixture where appropriate.
[0117] "Diastereoisomers" are stereoisomers that have at least two
asymmetric atoms, but which are not mirror-images of each
other.
[0118] The absolute stereochemistry is specified according to the
Cahn-Ingold-Prelog R--S system. When the compound is a pure
enantiomer the stereochemistry at each chiral carbon may be
specified by either R or S. Resolved compounds whose absolute
configuration is unknown are designated (+) or (-) depending on the
direction (dextro- or laevorotary) which they rotate the plane of
polarized light at the wavelength of the sodium D line.
[0119] The term "therapeutically effective amount" refers to that
amount of a compound of Formula I that is sufficient to effect
treatment, as defined below, when administered to a mammal in need
of such treatment. The therapeutically effective amount will vary
depending upon the subject and disease condition being treated, the
weight and age of the subject, the severity of the disease
condition, the manner of administration and the like, which can
readily be determined by one of ordinary skill in the art.
[0120] The term "treatment" or "treating" means any treatment of a
disease in a mammal, including: [0121] (i) preventing the disease,
that is, causing the clinical symptoms of the disease not to
develop; [0122] (ii) inhibiting the disease, that is, arresting the
development of clinical symptoms; and/or [0123] (iii) relieving the
disease, that is, causing the regression of clinical symptoms.
[0124] In many cases, the compounds of this invention are capable
of forming acid and/or base salts by virtue of the presence of
amino and/or carboxyl groups or groups similar thereto. The term
"pharmaceutically acceptable salt" refers to salts that retain the
biological effectiveness and properties of the compounds of Formula
I, and which are not biologically or otherwise undesirable.
Pharmaceutically acceptable base addition salts can be prepared
from inorganic and organic bases. Salts derived from inorganic
bases, include by way of example only, sodium, potassium, lithium,
ammonium, calcium and magnesium salts. Salts derived from organic
bases include, but are not limited to, salts of primary, secondary
and tertiary amines, such as alkyl amines, dialkyl amines, trialkyl
amines, substituted alkyl amines, di(substituted alkyl) amines,
tri(substituted alkyl) amines, alkenyl amines, dialkenyl amines,
trialkenyl amines, substituted alkenyl amines, di(substituted
alkenyl) amines, tri(substituted alkenyl) amines, cycloalkyl
amines, di(cycloalkyl) amines, tri(cycloalkyl) amines, substituted
cycloalkyl amines, disubstituted cycloalkyl amine, trisubstituted
cycloalkyl amines, cycloalkenyl amines, di(cycloalkenyl) amines,
tri(cycloalkenyl) amines, substituted cycloalkenyl amines,
disubstituted cycloalkenyl amine, trisubstituted cycloalkenyl
amines, aryl amines, diaryl amines, triaryl amines, heteroaryl
amines, diheteroaryl amines, triheteroaryl amines, heterocyclic
amines, diheterocyclic amines, triheterocyclic amines, mixed di-
and tri-amines where at least two of the substituents on the amine
are different and are selected from the group consisting of alkyl,
substituted alkyl, alkenyl, substituted alkenyl, cycloalkyl,
substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl,
aryl, heteroaryl, heterocyclic, and the like. Also included are
amines where the two or three substituents, together with the amino
nitrogen, form a heterocyclic or heteroaryl group.
[0125] Specific examples of suitable amines include, by way of
example only, isopropylamine, trimethyl amine, diethyl amine,
tri(iso-propyl) amine, tri(n-propyl) amine, ethanolamine,
2-dimethylaminoethanol, tromethamine, lysine, arginine, histidine,
caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine,
glucosamine, N-alkylglucamines, theobromine, purines, piperazine,
piperidine, morpholine, N-ethylpiperidine, and the like.
[0126] Pharmaceutically acceptable acid addition salts may be
prepared from inorganic and organic acids. Salts derived from
inorganic acids include hydrochloric acid, hydrobromic acid,
sulfuric acid, nitric acid, phosphoric acid, and the like. Salts
derived from organic acids include acetic acid, propionic acid,
glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid,
succinic acid, maleic acid, fumaric acid, tartaric acid, citric
acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic
acid, ethanesulfonic acid, p-toluene-sulfonic acid, salicylic acid,
and the like.
[0127] As used herein, "pharmaceutically acceptable carrier"
includes any and all solvents, dispersion media, coatings,
antibacterial and antifungal agents, isotonic and absorption
delaying agents and the like. The use of such media and agents for
pharmaceutically active substances is well known in the art. Except
insofar as any conventional media or agent is incompatible with the
active ingredient, its use in the therapeutic compositions is
contemplated. Supplementary active ingredients can also be
incorporated into the compositions.
Nomenclature
[0128] The naming and numbering of the compounds of the invention
is illustrated with a representative compound of Formula I in which
R.sup.1 is n-propyl, R.sup.2 is ethyl, X is 1,4-pyrazolenyl, Y is
--CH.sub.2--, and Z is 3-trifluoromethylyphenyl);
##STR00041##
which is named: [0129]
3-ethyl-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,-
3,7-trihydropurine-2,6-dione.
Synthetic Reaction Parameters
[0130] The terms "solvent", "inert organic solvent" or "inert
solvent" mean a solvent inert under the conditions of the reaction
being described in conjunction therewith [including, for example,
benzene, toluene, acetonitrile, tetrahydrofuran ("THF"),
dimethylformamide ("DMF"), chloroform, methylene chloride (or
dichloromethane), diethyl ether, methanol, pyridine and the like].
Unless specified to the contrary, the solvents used in the
reactions of the present invention are inert organic solvents, and
the reactions are carried out under an inert gas, preferably
nitrogen.
[0131] The term "q.s." means adding a quantity sufficient to
achieve a stated function, e.g., to bring a solution to the desired
volume (i.e., 100%).
Preparation of a Compound of Formula I
[0132] One preferred method of preparing compounds of Formula I is
shown in Reaction Scheme I.
##STR00042##
Step 1--Preparation of Formula (2)
[0133] The compound of formula (2) is made from the compound of
formula (1) by a reduction step. Conventional reducing techniques
may be used, for example using sodium dithionite in aqueous ammonia
solution; preferably reduction is carried out with hydrogen and a
metal catalyst. The reaction is carried out at in an inert solvent,
for example methanol, in the presence of a catalyst, for example
10% palladium on carbon catalyst, under an atmosphere of hydrogen,
preferably under pressure, for example at about 30 psi, for about 2
hours. When the reaction is substantially complete, the product of
formula (2) is isolated by conventional means tp provide a compound
of formula (2).
Step 2--Preparation of Formula (3)
[0134] The compound of formula (2) is then reacted with a
carboxylic acid of the formula Z-Y--X--CO.sub.2H in the presence of
a carbodiimide, for example
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride. The
reaction is conducted in a protic solvent, for example methanol,
ethanol, propanol, and the like, preferably methanol, at a
temperature of about 20-30.degree. C., preferably about room
temperature, for about 12-48 hours, preferably about 16 hours. When
the reaction is substantially complete, the product of formula (3)
is isolated conventionally, for example by removal of the solvent
under reduced pressure, and washing the product. Alternatively, the
next step can be carried out without any further purification.
Alternative Preparation of a Compound of Formula (3)
[0135] Alternatively, the carboxylic acid of the formula
Z-Y--X--CO.sub.2H is first converted to an acid halide of the
formula Z-Y--X--C(O)L, where L is chloro or bromo, by reacting with
a halogenating agent, for example thionyl chloride or thionyl
bromide, preferably thiony chloride. Alternatively, oxalyl
chloride, phosphorus pentachloride or phosphorus oxychloride may be
used. The reaction is preferably conducted in the absence of a
solvent, using excess halogenating agent, for example at a
temperature of about 60-80.degree. C., preferably about 70.degree.
C., for about 1-8 hours, preferably about 4 hours. When the
reaction is substantially complete, the product of formula
Z-Y--X--C(O)L is isolated conventionally, for example by removal of
the excess halogenating agent under reduced pressure.
[0136] The product is then reacted with a compound of formula (2)
in an inert solvent, for example acetonitrile, in the presence of a
tertiary base, for example triethylamine. The reaction is conducted
at an initial temperature of about 0 C, and then allowed to warm to
20-30.degree. C., preferably about room temperature, for about
12-48 hours, preferably about 16 hours. When the reaction is
substantially complete, the product of formula (3) is isolated
conventionally, for example by diluting the reaction mixture with
water, filtering off the product, and washing the product with
water followed by ether.
Step 3--Preparation of Formula I
[0137] The compound of formula (3) is then converted into a
compound of Formula I by a cyclization reaction. The reaction is
conducted in a protic solvent, for example methanol, ethanol,
propanol, and the like, preferably methanol, in the presence of a
base, for example potassium hydroxide, sodium hydroxide, sodium
methoxide, sodium ethoxide, potassium t-butoxide, preferably
aqueous sodium hydroxide, at a temperature of about 50-80.degree.
C., preferably about 80.degree. C., for about 1-8 hours, preferably
about 3 hours. When the reaction is substantially complete, the
product of Formula I is isolated conventionally, for example by
removal of the solvent under reduced pressure, acidifying the
residue with an aqueous acid, filtering off the product, then
washing and drying the product.
[0138] The compound of formula (1) may be prepared by various
methods. One preferred method is shown in Reaction Scheme II.
##STR00043##
Step 1--Preparation of Formula (5)
[0139] The compound of formula (4) is either commercially available
or prepared by means well known in the art. It is reacted with
ethyl cyanoacetate in a protic solvent, for example ethanol in the
presence of a strong base, for example sodium ethoxide. The
reaction is carried out at about reflux temperature, for about 4 to
about 24 hours. When the reaction is substantially complete, the
compound of formula (5) thus produced is isolated
conventionally.
Step 2 and 3--Preparation of Formula (7)
[0140] The compound of formula (5) is reacted with the
dimethylacetal of N,N-dimethylformamide in a polar solvent, for
example N,N-dimethylformamide. The reaction is carried out at about
40.degree. C., for about 1 hour. When the reaction is substantially
complete, the compound of formula (6) thus produced is reacted with
a compound of formula R.sup.1Hal, where Hat is chloro, bromo, or
iodo, in the presence of a base, for example potassium carbonate.
The reaction is carried out at about 80.degree. C., for about 4-24
hour. When the reaction is substantially complete, the product of
formula (7) is isolated conventionally, for example by evaporation
of the solvents under reduced pressure, and the residue is used in
the next reaction with no further purification.
Step 4--Preparation of Formula (8)
[0141] The compound of formula (7) is reacted with aqueous ammonia
in a polar solvent, for example suspended in methanol. The reaction
is carried out at about room temperature, for about 1-3 days. When
the reaction is substantially complete, the product of formula (8)
is isolated conventionally, for example by chromatography over a
silica gel column, eluting, for example, with a mixture of
dichloromethane/methanol.
Step 5--Preparation of Formula (1)
[0142] The compound of formula (8) is then mixed with sodium
nitrite in an aqueous acidic solvent, preferably acetic acid and
water, for example 50% acetic acid/water. The reaction is carried
out at a temperature of about 50-90.degree. C., preferably about
70.degree. C., for about 1 hour. When the reaction is substantially
complete, the product of formula (1) is isolated by conventional
means.
[0143] Alternatively, the reaction may be conducted in an aqueous
solvent, for example dimethylformamide and water, and reacted with
a strong acid, for example hydrochloric acid.
[0144] A compound of formula (8) can be prepared from a compound of
formula (10) using a similar method, as shown in Reaction Scheme
IIA.
##STR00044##
Step 2 and 3--Preparation of Formula (7)
[0145] The compound of formula (10) is reacted with the
dimethylacetal of N,N-dimethylformamide in a polar solvent, for
example N,N-dimethylformamide. The reaction is carried out at about
40.degree. C., for about 1 hour. When the reaction is substantially
complete, the compound of formula (6a) thus produced is reacted
with a compound of formula R.sup.2Hal, where Hal is chloro, bromo,
or iodo, in the presence of a base, for example potassium
carbonate. The reaction is carried out at about 80.degree. C., for
about 4-24 hour. When the reaction is substantially complete, the
product of formula (7) is isolated conventionally, for example by
evaporation of the solvents under reduced pressure, and the residue
is used in the next reaction with no further purification.
Step 4--Preparation of Formula (8)
[0146] The compound of formula (7) is reacted with aqueous ammonia
in a polar solvent, for example suspended in methanol. The reaction
is carried out at about room temperature, for about 1-3 days. When
the reaction is substantially complete, the product of formula (8)
is isolated conventionally, for example by chromatography over a
silica gel column, eluting, for example, with a mixture of
dichloromethane/methanol.
[0147] The compound of formula (3) may also be prepared by various
methods. One preferred method is shown in Reaction Scheme III.
##STR00045##
Step 1--Preparation of Formula (10)
[0148] The commercially available compound 6-aminouracil is first
silylated, for example by reaction with excess hexamethyldisilazane
as a solvent in the presence of a catalyst, for example ammonium
sulfate. The reaction is carried out at about reflux temperature,
for about 1-10 hours. When the reaction is substantially complete,
the silylated compound thus produced is isolated conventionally,
and then reacted with a compound of formula R.sup.1Hal, where Hal
is chloro, bromo, or iodo, preferably in the absence of a solvent.
The reaction is carried out at about reflux, for about 4-48 hours,
preferably about 12-16 hours. When the reaction is substantially
complete, the product of formula (10) is isolated by conventional
means.
Step 2--Preparation of Formula (11)
[0149] The compound of formula (10) is then dissolved in an aqueous
acid, for example aqueous acetic acid, and reacted with sodium
nitrite. The reaction is carried out at a temperature of about
20-50.degree. C., preferably about 30.degree. C., over about 30
minutes. When the reaction is substantially complete, the product
of formula (11) is isolated by conventional means, for example by
filtration.
Step 3--Preparation of Formula (12)
[0150] The compound of formula (11) is then reduced to a diamino
derivative. In general, the compound of formula (11) is dissolved
in aqueous ammonia, and then a reducing agent, for example sodium
hydrosulfite, added. The reaction is conducted at a temperature of
about 70.degree. C. When the reaction is substantially complete,
the product of formula (12) is isolated conventionally, for example
by filtration of the cooled reaction mixture.
Step 4--Preparation of Formula (13)
[0151] The compound of formula (12) is then reacted with a
carboxylic acid of the formula Z-Y--X--CO.sub.2H in the presence of
a carbodiimide, for example
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride. The
reaction is conducted at a temperature of about 20-30.degree. C.,
for about 12-48 hours. When the reaction is substantially complete,
the product of formula (13) is isolated conventionally, for example
by filtration of the cooled reaction mixture.
[0152] Alternatively, the carboxylic acid of the formula
Z-Y--X--CO.sub.2H is converted to an acid halide of the formula
Z-Y--X--C(O)L, where L is chloro or bromo, by reacting with a
halogenating agent, for example thionyl chloride or thionyl
bromide; alternatively, phosphorus pentachloride or phosphorus
oxychloride may be used. The reaction is preferably conducted in
the absence of a solvent, using excess halogenating agent, for
example at a temperature of about 60-80.degree. C., preferably
about 70.degree. C., for about 1-8 hours, preferably about 4 hours.
When the reaction is substantially complete, the product of formula
Z-Y--X--C(O)L is isolated conventionally, for example by removal of
the excess halogenating agent under reduced pressure.
[0153] The product of the formula Z-Y--X--C(O)L is then reacted
with a compound of formula (12) in an inert solvent, for example
acetonitrile, in the presence of a tertiary base, for example
triethylamine. The reaction is conducted at an initial temperature
of about 0 C, and then allowed to warm to 20-30.degree. C.,
preferably about room temperature, for about 12-48 hours,
preferably about 16 hours. When the reaction is substantially
complete, the product of formula (13) is isolated conventionally,
for example by diluting the reaction mixture with water, filtering
off the product, and washing the product with water followed by
ether.
Step 5--Preparation of Formula (3)
[0154] The compound of formula (13) is reacted with a compound of
formula R.sup.2Hal, where Hal is chloro, bromo, or iodo, in the
presence of a base, for example potassium carbonate. The reaction
is carried out at about room temperature, for about 4-24 hour,
preferably about 16 hours. When the reaction is substantially
complete, the product of formula (3) is isolated conventionally,
for example by evaporation of the solvents under reduced pressure,
and the residue may be purified conventionally, or may be used in
the next reaction with no further purification.
[0155] Another method of preparing a compound of formula (3) is
shown in Reaction Scheme IV.
##STR00046##
Step 1--Preparation of Formula (14)
[0156] The compound of formula (5) is then mixed with sodium
nitrite in an aqueous acidic solvent, preferably acetic acid and
water, for example 50% acetic acid/water. The reaction is carried
out at a temperature of about 50-90.degree. C., preferably about
70.degree. C., for about 1 hour. When the reaction is substantially
complete, the product of formula (14) is isolated by conventional
means.
[0157] Alternatively, the reaction may be conducted in an aqueous
solvent, for example dimethylformamide and water, and reacted with
a strong acid, for example hydrochloric acid.
Step 3--Preparation of Formula (15)
[0158] The compound of formula (14) is then reduced to a diamino
derivative. In general, the compound of formula (14) is dissolved
in aqueous ammonia, and then a reducing agent, for example sodium
hydrosulfite, added. The reaction is conducted at a temperature of
about 70.degree. C. When the reaction is substantially complete,
the product of formula (15) is isolated conventionally, for example
by filtration of the cooled reaction mixture.
Step 4--Preparation of Formula (16)
[0159] The compound of formula (15) is then reacted with a
carboxylic acid of the formula Z-Y--X--CO.sub.2 H in the presence
of a carbodiimide, for example
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride. The
reaction is conducted at a temperature of about 20-30.degree. C.,
for about 12-48 hours, in an inert solvent, for example methanol.
When the reaction is substantially complete, the product of formula
(16) is isolated conventionally, for example by filtration of the
cooled reaction mixture.
[0160] Alternatively, the carboxylic acid of the formula
Z-Y--X--CO.sub.2H is converted to an acid halide of the formula
Z-Y--X--C(O)L, where L is chloro or bromo, by reacting with a
halogenating agent, for example thionyl chloride or thionyl
bromide; alternatively, phosphorus pentachloride or phosphorus
oxychloride may be used. The reaction is preferably conducted in
the absence of a solvent, using excess halogenating agent, for
example at a temperature of about 60-80.degree. C., preferably
about 70.degree. C., for about 1-8 hours, preferably about 4 hours.
When the reaction is substantially complete, the product of formula
Z-Y--X--C(O)L is isolated conventionally, for example by removal of
the excess halogenating agent under reduced pressure.
[0161] The product of the formula Z-Y--X--C(O)L is then reacted
with a compound of formula (15) in an inert solvent, for example
acetonitrile, in the presence of a tertiary base, for example
triethylamine. The reaction is conducted at an initial temperature
of about 0 C, and then allowed to warm to 20-30.degree. C.,
preferably about room temperature, for about 12-48 hours,
preferably about 16 hours. When the reaction is substantially
complete, the product of formula (16) is isolated conventionally,
for example by diluting the reaction mixture with water, filtering
off the product, and washing the product with water followed by
ether.
Step 5--Preparation of Formula (3)
[0162] The compound of formula (16) is reacted with a compound of
formula R.sup.1Hal, where Hal is chloro, bromo, or iodo, in the
presence of a base, for example potassium carbonate. The reaction
is carried out at about 80.degree. C., for about 4-24 hour,
preferably about 16 hours. When the reaction is substantially
complete, the product of formula (3) is isolated conventionally,
for example by evaporation of the solvents under reduced pressure,
and the residue may be purified conventionally, or may be used in
the next reaction with no further purification.
[0163] An example of a synthesis of a compound of Z-Y--X--CO.sub.2H
in which X is pyrazol-1,4-yl, Y is methylene, and Z is
3-trifluoromethylphenyl, is shown in Reaction Scheme V.
##STR00047##
[0164] Ethyl pyrazole-4-carboxylate is reacted with
1-(bromomethyl)-3-(trifluoromethyl)benzene in acetone in the
presence of potassium carbonate. The product, ethyl
1-{[3-(trifluoromethyl)phenyl]methyl}pyrazole-4-carboxylate, is
then hydrolyzed with potassium hydroxide in methanol, to provide
1-{[3-(trifluoromethyl)phenyl]methyl}pyrazole-4-carboxylic
acid.
Utility, Testing and Administration
General Utility
[0165] The compounds of Formula I are effective in the treatment of
conditions that respond to administration of A.sub.2B adenosine
receptor antagonists. Such conditions include, but are not limited
to, at least one of diarrhea, atherosclerosis, restenosis,
rheumatoid arthritis, diabetes, in particular type-II diabetes,
macular degeneration, diabetic retinopathy, cancer, senile
dementia, Alzheimer's disease, Parkinson's disease, traumatic brain
injury, and Type I hypersensitivity reactions, including asthma,
atopic eczema, and hay fever.
Testing
[0166] Activity testing is conducted as described in those patents
and patent applications referenced above, and in the Examples
below, and by methods apparent to one skilled in the art.
Pharmaceutical Compositions
[0167] The compounds of Formula I are usually administered in the
form of pharmaceutical compositions. This invention therefore
provides pharmaceutical compositions that contain, as the active
ingredient, one or more of the compounds of Formula I, or a
pharmaceutically acceptable salt or ester thereof, and one or more
pharmaceutically acceptable excipients, carriers, including inert
solid diluents and fillers, diluents, including sterile aqueous
solution and various organic solvents, permeation enhancers,
solubilizers and adjuvants. The compounds of Formula I may be
administered alone or in combination with other therapeutic agents.
Such compositions are prepared in a manner well known in the
pharmaceutical art (see, e.g., Remington's Pharmaceutical Sciences,
Mace Publishing Co., Philadelphia, Pa. 17.sup.th Ed. (1985) and
"Modern Pharmaceutics", Marcel Dekker, Inc. 3.sup.rd Ed. (G. S.
Banker & C. T. Rhodes, Eds.).
Administration
[0168] The compounds of Formula I may be administered in either
single or multiple doses by any of the accepted modes of
administration of agents having similar utilities, for example as
described in those patents and patent applications incorporated by
reference, including rectal, buccal, intranasal and transdermal
routes, by intra-arterial injection, intravenously,
intraperitoneally, parenterally, intramuscularly, subcutaneously,
orally, topically, as an inhalant, or via an impregnated or coated
device such as a stent, for example, or an artery-inserted
cylindrical polymer.
[0169] One mode for administration is parental, particularly by
injection. The forms in which the novel compositions of the present
invention may be incorporated for administration by injection
include aqueous or oil suspensions, or emulsions, with sesame oil,
corn oil, cottonseed oil, or peanut oil, as well as elixirs,
mannitol, dextrose, or a sterile aqueous solution, and similar
pharmaceutical vehicles. Aqueous solutions in saline are also
conventionally used for injection, but less preferred in the
context of the present invention. Ethanol, glycerol, propylene
glycol, liquid polyethylene glycol, and the like (and suitable
mixtures thereof), cyclodextrin derivatives, and vegetable oils may
also be employed. The proper fluidity can be maintained, for
example, by the use of a coating, such as lecithin, by the
maintenance of the required particle size in the case of dispersion
and by the use of surfactants. The prevention of the action of
microorganisms can be brought about by various antibacterial and
antifungal agents, for example, parabens, chlorobutanol, phenol,
sorbic acid, thimerosal, and the like.
[0170] Sterile injectable solutions are prepared by incorporating
the compound of Formula I in the required amount in the appropriate
solvent with various other ingredients as enumerated above, as
required, followed by filtered sterilization. Generally,
dispersions are prepared by incorporating the various sterilized
active ingredients into a sterile vehicle which contains the basic
dispersion medium and the required other ingredients from those
enumerated above. In the case of sterile powders for the
preparation of sterile injectable solutions, the preferred methods
of preparation are vacuum-drying and freeze-drying techniques which
yield a powder of the active ingredient plus any additional desired
ingredient from a previously sterile-filtered solution thereof.
[0171] Oral administration is another route for administration of
the compounds of Formula I. Administration may be via capsule or
enteric coated tablets, or the like. In making the pharmaceutical
compositions that include at least one compound of Formula I, the
active ingredient is usually diluted by an excipient and/or
enclosed within such a carrier that can be in the form of a
capsule, sachet, paper or other container. When the excipient
serves as a diluent, in can be a solid, semi-solid, or liquid
material (as above), which acts as a vehicle, carrier or medium for
the active ingredient. Thus, the compositions can be in the form of
tablets, pills, powders, lozenges, sachets, cachets, elixirs,
suspensions, emulsions, solutions, syrups, aerosols (as a solid or
in a liquid medium), ointments containing, for example, up to 10%
by weight of the active compound, soft and hard gelatin capsules,
sterile injectable solutions, and sterile packaged powders.
[0172] Some examples of suitable excipients include lactose,
dextrose, sucrose, sorbitol, mannitol, starches, gum acacia,
calcium phosphate, alginates, tragacanth, gelatin, calcium
silicate, microcrystalline cellulose, polyvinylpyrrolidone,
cellulose, sterile water, syrup, and methyl cellulose. The
formulations can additionally include: lubricating agents such as
talc, magnesium stearate, and mineral oil; wetting agents;
emulsifying and suspending agents; preserving agents such as
methyl- and propylhydroxy-benzoates; sweetening agents; and
flavoring agents.
[0173] The compositions of the invention can be formulated so as to
provide quick, sustained or delayed release of the active
ingredient after administration to the patient by employing
procedures known in the art. Controlled release drug delivery
systems for oral administration include osmotic pump systems and
dissolutional systems containing polymer-coated reservoirs or
drug-polymer matrix formulations. Examples of controlled release
systems are given in U.S. Pat. Nos. 3,845,770; 4,326,525;
4,902,514; and 5,616,345. Another formulation for use in the
methods of the present invention employs transdermal delivery
devices ("patches"). Such transdermal patches may be used to
provide continuous or discontinuous infusion of the compounds of
the present invention in controlled amounts. The construction and
use of transdermal patches for the delivery of pharmaceutical
agents is well known in the art. See, e.g., U.S. Pat. Nos.
5,023,252, 4,992,445 and 5,001,139. Such patches may be constructed
for continuous, pulsatile, or on demand delivery of pharmaceutical
agents.
[0174] The compositions are preferably formulated in a unit dosage
form. The term "unit dosage forms" refers to physically discrete
units suitable as unitary dosages for human subjects and other
mammals, each unit containing a predetermined quantity of active
material calculated to produce the desired therapeutic effect, in
association with a suitable pharmaceutical excipient (e.g., a
tablet, capsule, ampoule). The compounds of Formula I are effective
over a wide dosage range and is generally administered in a
pharmaceutically effective amount. Preferably, for oral
administration, each dosage unit contains from 10 mg to 2 g of a
compound of Formula I, more preferably from 10 to 700 mg, and for
parenteral administration, preferably from 10 to 700 mg of a
compound of Formula I, more preferably about 50-200 mg. It will be
understood, however, that the amount of the compound of Formula I
actually administered will be determined by a physician, in the
light of the relevant circumstances, including the condition to be
treated, the chosen route of administration, the actual compound
administered and its relative activity, the age, weight, and
response of the individual patient, the severity of the patient's
symptoms, and the like.
[0175] For preparing solid compositions such as tablets, the
principal active ingredient is mixed with a pharmaceutical
excipient to form a solid preformulation composition containing a
homogeneous mixture of a compound of the present invention. When
referring to these preformulation compositions as homogeneous, it
is meant that the active ingredient is dispersed evenly throughout
the composition so that the composition may be readily subdivided
into equally effective unit dosage forms such as tablets, pills and
capsules.
[0176] The tablets or pills of the present invention may be coated
or otherwise compounded to provide a dosage form affording the
advantage of prolonged action, or to protect from the acid
conditions of the stomach. For example, the tablet or pill can
comprise an inner dosage and an outer dosage component, the latter
being in the form of an envelope over the former. The two
components can be separated by an enteric layer that serves to
resist disintegration in the stomach and permit the inner component
to pass intact into the duodenum or to be delayed in release. A
variety of materials can be used for such enteric layers or
coatings, such materials including a number of polymeric acids and
mixtures of polymeric acids with such materials as shellac, cetyl
alcohol, and cellulose acetate.
[0177] Compositions for inhalation or insufflation include
solutions and suspensions in pharmaceutically acceptable, aqueous
or organic solvents, or mixtures thereof, and powders. The liquid
or solid compositions may contain suitable pharmaceutically
acceptable excipients as described supra. Preferably the
compositions are administered by the oral or nasal respiratory
route for local or systemic effect. Compositions in preferably
pharmaceutically acceptable solvents may be nebulized by use of
inert gases. Nebulized solutions may be inhaled directly from the
nebulizing device or the nebulizing device may be attached to a
face mask tent, or intermittent positive pressure breathing
machine. Solution, suspension, or powder compositions may be
administered, preferably orally or nasally, from devices that
deliver the formulation in an appropriate manner.
[0178] The following examples are included to demonstrate preferred
embodiments of the invention. It should be appreciated by those of
skill in the art that the techniques disclosed in the examples
which follow represent techniques discovered by the inventor to
function well in the practice of the invention, and thus can be
considered to constitute preferred modes for its practice. However,
those of skill in the art should, in light of the present
disclosure, appreciate that many changes can be made in the
specific embodiments which are disclosed and still obtain a like or
similar result without departing from the spirit and scope of the
invention.
EXAMPLE 1
Preparation of a Compound of Formula (5)
[0179] A. Preparation of a Compound of Formula (5) in which R.sup.2
is Ethyl
##STR00048##
[0180] A solution of sodium ethoxide was prepared from sodium (4.8
g, 226 mmol) and dry ethanol (150 ml). To this solution was added
amino-N-ethylamide (10 g, 113 m mol) and ethyl cyanoacetate (12.8
g, 113 mmol). This reaction mixture was stirred at reflux for 6
hours, cooled, and solvent removed from the reaction mixture under
reduced pressure. The residue was dissolved in water (50 ml), and
the pH adjusted to 7 with hydrochloric acid. The mixture was
allowed to stand overnight at 0.degree. C., and the precipitate
filtered off, washed with water and air-dried, to provide
6-amino-1-ethyl-1,3-dihydropymmidine-2,4-dione, a compound of
formula (5).
[0181] .sup.1H-NMR (DMSO-d6) .delta. 10.29 (s, 1H), 6.79 (s, 2H),
4.51 (s, 1H), 3.74-3.79 (m, 2H), 1.07 (t, 3H, J=7.03 Hz); MS m/z
155.98 (M.sup.+), 177.99 (M.sup.++Na)
B. Preparation of a Compound of Formula (5) in which R.sup.2 is
Methyl
[0182] Similarly, following the procedure of Example 1A, but
replacing amino-N-ethylamide with amino-N-methylamide,
6-amino-1-methyl-1,3-dihydropyrimidine-2,4-dione was prepared.
C. Preparation of a Compound of Formula (5) Varying R.sup.2
[0183] Similarly, following the procedure of Example 1A, but
replacing amino-N-ethylamide with other compounds of formula (4),
other compounds of formula (5) are prepared.
EXAMPLE 2
Preparation of a Compound of Formula (6)
[0184] A. Preparation of a Compound of Formula (6) in which R.sup.2
is Ethyl
##STR00049##
[0185] A suspension of
6-amino-1-ethyl-1,3-dihydropyrimidine-2,4-dione (0.77 g, 5 mmol) in
anhydrous N,N-dimethylacetamide (25 ml) and N,N-dimethylformamide
dimethylacetal (2.7 ml, 20 mmol) and was warmed at 40.degree. C.
for 90 minutes. Solvent was then removed under reduced pressure,
and the residue triturated with ethanol, filtered, and washed with
ethanol, to provide
6-[2-(dimethylamino)-1-azavinyl]-1-ethyl-1,3-dihydropyrimidine-2,4-dione,
a compound of formula (6). .sup.1H-NMR (DMSO-d6) .delta. 10.62 (s,
1H), 8.08 (s, 1H), 4.99 (s, 1H), 3.88-3.95 (m, 2H), 3.13 (s, 3H),
2.99 (s, 3H), 1.07 (t, 3H, J=7.03 Hz); MS m/z 210.86 (M.sup.+),
232.87 (M.sup.++Na)
B. Preparation of a Compound of Formula (6) in which R.sup.2 is
Methyl
[0186] Similarly, following the procedure of Example 2A, but
replacing 6-amino-1-ethyl-1,3-dihydropyrimidine-2,4-dione with
6-amino-1-methyl-1,3-dihydropyrimidine-2,4-dione,
6-[2-(dimethylamino)-1-azavinyl]-1-methyl-1,3-dihydropyrimidine-2,4-dione
was prepared.
C. Preparation of a Compound of Formula (6) Varying R.sup.2
[0187] Similarly, following the procedure of Example 2A, but
replacing 6-amino-1-ethyl-1,3-dihydropyrimidine-2,4-dione with
other compounds of formula (5), other compounds of formula (6) are
prepared.
EXAMPLE 3
Preparation of a Compound of Formula (7)
[0188] A. Preparation of a Compound of Formula (7) in which R.sup.1
is n-Propyl and R.sup.2 is Ethyl
##STR00050##
[0189] A mixture of a solution of
6-[2-(dimethylamino)-1-azavinyl]-1-ethyl-1,3-dihydropyrimidine-2,4-dione
(1.5 g, 7.1 mmol) in dimethylformamide (25 ml), potassium carbonate
(1.5 g, 11 mmol) and n-propyl iodide (1.54 g, 11 mmol) was stirred
at 80.degree. C. for 5 hours. The reaction mixture was cooled to
room temperature, filtered, the solvents were evaporated and the
product of formula (7),
6-[2-(dimethylamino)-1-azavinyl]-1-ethyl-3-propyl-1,3-dihydropyrimidine-2-
,4-dione, was used as such in the next reaction.
B. Preparation of a Compound of Formula (7), Varying R.sup.1 and
R.sup.2
[0190] Similarly, following the procedure of Example 3A, but
replacing
6-[2-(dimethylamino)-1-azavinyl]-1-ethyl-1,3-dihydropyrimidine-2,4-dione
with other compounds of formula (6), the following compounds of
formula (7) were prepared: [0191]
6-[2-(dimethylamino)-1-azavinyl]-1-methyl-3-propyl-1,3-dihydropyrimidine--
2,4-dione. [0192]
6-[2-(dimethylamino)-1-azavinyl]-1-methyl-3-cyclopropylmethyl-1,3-dihydro-
pyrimidine-2,4-dione; [0193]
6-[2-(dimethylamino)-1-azavinyl]-1-ethyl-3-cyclopropylmethyl-1,3-dihydrop-
yrimidine-2,4-dione; [0194]
6-[2-(dimethylamino)-1-azavinyl]-1-methyl-3-(2-methylpropyl)-1,3-dihydrop-
yrimidine-2,4-dione; and [0195]
6-[2-(dimethylamino)-1-azavinyl]-1-ethyl-3-(2-methylpropyl)-1,3-dihydropy-
rimidine-2,4-dione.
C. Preparation of a Compound of Formula (7), Varying R.sup.1 and
R.sup.2
[0196] Similarly, following the procedure of Example 3A, but
replacing
6-[2-(dimethylamino)-1-azavinyl]-1-ethyl-1,3-dihydropyrimidine-2,4-dione
with other compounds of formula (6), other compounds of formula (7)
are prepared.
EXAMPLE 4
Preparation of a Compound of Formula (8)
[0197] A. Preparation of a Compound of Formula (8) in which R.sup.1
is n-Propyl and R.sup.2 is Ethyl
##STR00051##
[0198] A solution of
6-[2-(dimethylamino)-1-azavinyl]-1-ethyl-3-propyl-1,3-dihydropyrimidine-2-
,4-dione (2.1 g) was dissolved in a mixture of methanol (10 ml) and
28% aqueous ammonia solution (20 ml), and stirred for 72 hours at
room temperature. Solvent was then removed under reduced pressure,
and the residue purified by chromatography on a silica gel column,
eluting with a mixture of dichloromethane/methanol (15/1), to
provide 6-amino-1-ethyl-3-propyl-1,3-dihydropyrimidine-2,4-dione, a
compound of formula (8).
[0199] .sup.1H-NMR (DMSO-d6) .delta. 6.80 (s, 2H), 4.64 (s, 1H),
3.79-3.84 (m, 2H), 3.63-3.67 (m, 2H), 1.41-1.51 (m, 2H), 1.09 (t,
3H, J=7.03 Hz), 0.80 (t, 3H, J=7.42 Hz); MS m/z 197.82
(M.sup.+)
B. Preparation of a Compound of Formula (8), Varying R.sup.1 and
R.sup.2
[0200] Similarly, following the procedure of Example 4A, but
replacing
6-[2-(dimethylamino)-1-azavinyl]-1-ethyl-3-propyl-1,3-dihydropyrimidine-2-
,4-dione with other compounds of formula (7), the following
compounds of formula (8) were prepared: [0201]
6-amino-1-methyl-3-propyl-1,3-dihydropyrimidine-2,4-dione; [0202]
6-amino-1-methyl-3-cyclopropylmethyl-1,3-dihydropyrimidine-2,4-dione;
[0203]
6-amino-1-ethyl-3-cyclopropylmethyl-1,3-dihydropyrimidine-2,4-dion-
e; [0204]
6-amino-1-methyl-3-(2-methylpropyl)-1,3-dihydropyrimidine-2,4-di-
one; and [0205]
6-amino-1-ethyl-3-(2-methylpropyl)-1,3-dihydropyrimidine-2,4-dione.
C. Preparation of a Compound of Formula (7) Varying R.sup.1 and
R.sup.2
[0206] Similarly, following the procedure of Example 4A, but
replacing
6-[2-(dimethylamino)-1-azavinyl]-1-ethyl-3-propyl-1,3-dihydropyrimidine-2-
,4-dione with other compounds of formula (7), other compounds of
formula (8) are prepared.
EXAMPLE 5
Preparation of a Compound of Formula (1)
[0207] A. Preparation of a Compound of Formula (1) in which R.sup.1
is n-Propyl and R.sup.2 is Ethyl
##STR00052##
[0208] To a solution of
6-amino-1-ethyl-3-propyl-1,3-dihydropyrimidine-2,4-dione (1.4 g,
7.1 mmol) in a mixture of 50% acetic acid/water (35 ml) was added
sodium nitrite (2 g, 28.4 mmol) in portions over a period of 10
minutes. The mixture was stirred at 70.degree. C. for 1 hour, then
the reaction mixture concentrated to a low volume under reduced
pressure. The solid was filtered off, and washed with water, to
provide
6-amino-1-ethyl-5-nitroso-3-propyl-1,3-dihydropyrimidine-2,4-dione,
a compound of formula (1).
[0209] MS m/z 227.05 (M.sup.+), 249.08 (M.sup.++Na)
B. Preparation of a Compound of Formula (1), Varying R.sup.1 and
R.sup.2
[0210] Similarly, following the procedure of Example 5A, but
replacing 6-amino-1-ethyl-3-propyl-1,3-dihydropyrimidine-2,4-dione
with other compounds of formula (8), the following compounds of
formula (1) were prepared: [0211]
6-amino-1-methyl-5-nitroso-3-propyl-1,3-dihydropyrimidine-2,4-dione;
[0212]
6-amino-1-methyl-3-cyclopropylmethyl-5-nitroso-1,3-dihydropyrimidi-
ne-2,4-dione; [0213]
6-amino-1-ethyl-3-cyclopropylmethyl-5-nitroso-1,3-dihydropyrimidine-2,4-d-
ione; [0214]
6-amino-1-methyl-3-(2-methylpropyl)-5-nitroso-1,3-dihydropyrimidine-2,4-d-
ione; and [0215]
6-amino-1-ethyl-3-(2-methylpropyl)-5-nitroso-1,3-dihydropyrimidine-2,4-di-
one.
C. Preparation of a Compound of Formula (1) Varying R.sup.1 and
R.sup.2
[0216] Similarly, following the procedure of Example 5A, but
replacing 6-amino-1-ethyl-3-propyl-1,3-dihydropyrimidine-2,4-dione
with other compounds of formula (8), other compounds of formula (1)
are prepared.
EXAMPLE 6
Preparation of a Compound of Formula (2)
[0217] A. Preparation of a Compound of Formula (2) in which R.sup.1
is n-Propyl and R.sup.2 is Ethyl
##STR00053##
[0218] To a solution of
6-amino-1-ethyl-5-nitroso-3-propyl-1,3-dihydropyrimidine-2,4-dione
(300 mg) in methanol (10 ml) was added 10% palladium on carbon
catalyst (50 mg), and the mixture was hydrogenated under hydrogen
at 30 psi for 2 hours. The mixture was filtered through celite, and
solvent was removed from the filtrate under reduced pressure, to
provide
5,6-diamino-1-ethyl-3-propyl-1,3-dihydropyrimidine-2,4-dione, a
compound of formula (2).
[0219] MS m/z 213.03 (M.sup.+), 235.06 (M.sup.++Na)
B. Preparation of a Compound of Formula (2), Varying R.sup.1 and
R.sup.2
[0220] Similarly, following the procedure of Example 6A, but
replacing
6-amino-1-ethyl-5-nitroso-3-propyl-1,3-dihydropyrimidine-2,4-dione
with other compounds of formula (1), the following compounds of
formula (2) were prepared: [0221]
5,6-diamino-1-methyl-3-propyl-1,3-dihydropyrimidine-2,4-dione;
[0222]
5,6-diamino-1-methyl-3-cyclopropylmethyl-1,3-dihydropyrimidine-2,4-dione;
[0223]
5,6-diamino-1-ethyl-3-cyclopropylmethyl-1,3-dihydropyrimidine-2,4--
dione; [0224]
5,6-amino-1-methyl-3-(2-methylpropyl)-1,3-dihydropyrimidine-2,4-dione;
and [0225]
5,6-diamino-1-ethyl-3-(2-methylpropyl)-1,3-dihydropyrimidine-2,4-dione.
C. Preparation of a Compound of Formula (2) Varying R.sup.1 and
R.sup.2
[0226] Similarly, following the procedure of Example 6A, but
replacing
6-amino-1-ethyl-5-nitroso-3-propyl-1,3-dihydropyrimidine-2,4-dione
with other compounds of formula (1), other compounds of formula (2)
are prepared.
EXAMPLE 7
Preparation of a Compound of Formula (3)
[0227] A. Preparation of a Compound of Formula (3) in which R.sup.1
is n-Propyl, R.sup.2 is Ethyl, X is 1,4-Pyrazolyl, Y is Methylene,
and Z is 3-Trifluoromethylphenyl
##STR00054##
[0228] To a mixture of
5,6-diamino-1-ethyl-3-propyl-1,3-dihydropyrimidine-2,4-dione (100
mg, 0.47 mmol) and
1-{[3-(trifluoromethyl)phenyl]methyl}pyrazole-4-carboxylic acid
(0.151 g, 0.56 mmol) in methanol (10 ml) was added
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.135
g, 0.7 mmol), and the reaction mixture was stirred overnight at
room temperature. The solvent was removed under reduced pressure,
and the residue purified using Bistag, eluting with 10%
methanol/methylene chloride, to provide
N-(6-amino-1-ethyl-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[3-(-
trifluoromethyl)phenyl]methyl}-pyrazol-4-yl)carboxamide.
[0229] .sup.1H-NMR (DMSO-d6) .delta. 8.59 (s, 1H), 8.02 (s, 1H),
7.59-7.71 (m, 4H), 6.71 (s, 2H), 5.51 (s, 2H), 3.91-3.96 (m, 2H),
3.70-3.75 (m, 2H), 1.47-1.55 (m, 2H), 1.14 (t, 3H, J=7.03 Hz), 0.85
(t, 3H, J=7.42 Hz).
B. Preparation of a Compound of Formula (3), Varying R.sup.1,
R.sup.2, X, Y, and Z
[0230] Similarly, following the procedure of Example 7A or 7B, but
optionally replacing
5,6-diamino-1-ethyl-3-propyl-1,3-dihydropyrimidine-2,4-dione with
other compounds of formula (2), and optionally replacing
1-{[3-(trifluoromethyl)phenyl]methyl}pyrazole-4-carboxylic acid
with other compounds of formula Z-Y--X--CO.sub.2H, the following
compounds of formula (3) were prepared: [0231]
N-(6-amino-1-methyl-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[3--
(trifluoromethyl)phenyl]methyl}-pyrazol-4-yl)carboxamide; [0232]
N-(6-amino-1-methyl-2,4-dioxo-3-cyclopropylmethyl(1,3-dihydropyrimidin-5--
yl))(1-{[3-(trifluoromethyl)phenyl]methyl}-pyrazol-4-yl)carboxamide;
[0233]
N-(6-amino-1-ethyl-2,4-dioxo-3-cyclopropylmethyl(1,3-dihydropyrimi-
din-5-yl))(1-{[3-(trifluoromethyl)phenyl]methyl}-pyrazol-4-yl)carboxamide;
[0234]
N-(6-amino-1-methyl-2,4-dioxo-3-ethyl(1,3-dihydropyrimidin-5-yl))(-
1-{[3-fluorophenyl]methyl}-pyrazol-4-yl)carboxamide; [0235]
N-(6-amino-1-methyl-2,4-dioxo-3-cyclopropylmethyl(1,3-dihydropyrimidin-5--
yl))(1-{[3-fluorophenyl]methyl}-pyrazol-4-yl)carboxamide; [0236]
N-(6-amino-1-ethyl-2,4-dioxo-3-cyclopropylmethyl(1,3-dihydropyrimidin-5-y-
l))(1-{[3-fluorophenyl]methyl}-pyrazol-4-yl)carboxamide; [0237]
N-[6-amino-3-(cyclopropylmethyl)-1-methyl-2,4-dioxo(1,3-dihydropyrimidin--
5-yl)][1-benzylpyrazol-4-yl]carboxamide; [0238]
N-(6-amino-1-methyl-2,4-dioxo-3-cyclopropylmethyl(1,3-dihydropyrimidin-5--
yl))(1-{[3-cyanophenyl]methyl}-pyrazol-4-yl)carboxamide; [0239]
[1-(2-(1H-1,2,3,4-tetraazol-5-yl)ethyl)pyrazol-4-yl]-N-[6-amino-3-(cyclop-
ropylmethyl)-1-methyl-2,4-dioxo(1,3-dihydropyrimidin-5-yl)]carboxamide;
[0240]
N-[6-amino-3-(cyclopropylmethyl)-1-ethyl-2,4-dioxo(1,3-dihydropyri-
midin-5-yl)](1-{[6-(trifluoromethyl)(3-pyridyl)]methyl}pyrazol-4-yl)carbox-
amide; [0241]
N-[6-amino-3-propyl)-1-ethyl-2,4-dioxo(1,3-dihydropyrimidin-5-yl)](1-{(2--
pyridyl)]methyl}pyrazol-4-yl)carboxamide; [0242]
N-[6-amino-3-(2-methylpropyl)-1-methyl-2,4-dioxo(1,3-dihydropyrimidin-5-y-
l)]1-benzylpyrazol-4-yl]carboxamide; [0243]
N-[6-amino-3-(2-methylpropyl)-1-methyl-2,4-dioxo(1,3-dihydropyrimidin-5-y-
l)][1-{[3-fluorophenyl]methyl}pyrazol-4-yl]carboxamide; [0244]
N-[6-amino-3-(2-methylpropyl)-1-ethyl-2,4-dioxo(1,3-dihydropyrimidin-5-yl-
)][1-{[3-fluorophenyl]methyl}pyrazol-4-yl]carboxamide; [0245]
N-[6-amino-3-(2-methylpropyl)-1-methyl-2,4-dioxo(1,3-dihydropyrimidin-5-y-
l)][1-{[3-(trifluoromethyl)phenyl]methyl{pyrazol-4-yl]carboxamide;
and [0246]
N-[6-amino-3-(2-methylpropyl)-1-ethyl-2,4-dioxo(1,3-dihydropyrimid-
in-5-yl)]
(1-{[6-(trifluoromethyl)(3-pyridyl)]methyl}pyrazol-4-yl)carboxam-
ide.
C. Preparation of a Compound of Formula (2) Varying R.sup.1 and
R.sup.2
[0247] Similarly, following the procedure of Example 7A, but
optionally replacing
5,6-diamino-1-ethyl-3-propyl-1,3-dihydropyrimidine-2,4-dione with
other compounds of formula (2), and optionally replacing
1-{[3-(trifluoromethyl)phenyl]methyl}pyrazole-4-carboxylic acid
with other compounds of formula Z-Y--X--CO.sub.2H, other compounds
of formula (3) are prepared.
EXAMPLE 8
Preparation of a Compound of Formula I
[0248] A. Preparation of a Compound of Formula I in which R.sup.1
is n-Propyl, R.sup.2 is Ethyl, X is 1,4-Pyrazolyl, Y is Methylene,
and Z is 3-Trifluoromethylphenyl
##STR00055##
[0249] A mixture of
N-(6-amino-1-ethyl-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[3-(-
trifluoromethyl)phenyl]methyl}pyrazol-3-yl)carboxamide (80 mg, 0.17
mmol), 10% aqueous sodium hydroxide (5 ml), and methanol (5 ml) was
stirred at 100.degree. C. for 2 hours. The mixture was cooled,
methanol removed under reduced pressure, and the residue diluted
with water and acidified with hydrochloric acid. The precipitate
was filtered off, washed with water, then methanol, to provide
3-ethyl-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,-
3,7-trihydropurine-2,6-dione, a compound of Formula I.
[0250] .sup.1H-NMR (DMSO-d6) .delta. 8.57 (s, 1H), 8.15 (s, 1H),
7.60-7.75 (m, 4H), 5.54 (s, 2H), 4.05-4.50 (m, 2H), 3.87-3.91 (m,
2H), 1.55-1.64 (m, 2H), 1.25 (t, 3H, J=7.03 Hz), 0.90 (t, 3H,
J=7.42 Hz); MS m/z 447.2 (M.sup.+).
B. Preparation of a Compound of Formula I, Varying R.sup.1,
R.sup.2, X, Y, and Z
[0251] Similarly, following the procedure of Example 8A, but
replacing
N-(6-amino-1-ethyl-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[3-(-
trifluoromethyl)phenyl]-methyl}pyrazol-3-yl)carboxamide with other
compounds of formula (3), the following compounds of Formula I were
prepared: [0252]
1-cyclopropylmethyl-3-methyl-8-[1-(phenylmethyl)pyrazol-4-yl]-1,3,7-trihy-
dropurine-2,6-dione; [0253]
1-cyclopropylmethyl-3-methyl-8-{1-[(3-trifluoromethylphenyl)methyl]pyrazo-
l-4-yl}-1,3,7-trihydropurine-2,6-dione; [0254]
1-cyclopropylmethyl-3-ethyl-8-{1-[(3-trifluoromethylphenyl)methyl]pyrazol-
-4-yl}-1,3,7-trihydropurine-2,6-dione; [0255]
1-cyclopropylmethyl-3-methyl-8-{1-[(3-fluorophenyl)methyl]pyrazol-4-yl}-1-
,3,7-trihydropurine-2,6-dione; [0256]
1-cyclopropylmethyl-3-ethyl-8-{1-[(3-fluorophenyl)methyl]pyrazol-4-yl}-1,-
3,7-trihydropurine-2,6-dione; [0257]
1-cyclopropylmethyl-3-ethyl-8-(1-{[6-(trifluoromethyl)(3-pyridyl)]methyl}-
pyrazol-4-yl)-1,3,7-trihydropurine-2,6-dione; [0258]
3-({4-[1-(cyclopropylmethyl)-3-methyl-2,6-dioxo-1,3,7-trihydropurin-8-yl]-
pyrazolyl}methyl)benzenecarbonitrile; [0259]
8-[1-(2-(1H-1,2,3,4-tetraazol-5-yl)ethyl)pyrazol-4-yl]-3-methyl-1-cyclopr-
opylmethyl-1,3,7-trihydropurine-2,6-dione; [0260]
1-(2-methylpropyl)-3-methyl-8-[1-benzylpyrazol-4-yl]-1,3,7-trihydropurine-
-2,6-dione; [0261]
1-(2-methylpropyl)-3-ethyl-8-{1-[(3-fluorophenyl)methyl]pyrazol-4-yl}-1,3-
,7-trihydropurine-2,6-dione; [0262]
1-(2-methylpropyl)-3-methyl-8-{1-[(3-trifluoromethylphenyl)methyl]pyrazol-
-4-yl}-1,3,7-trihydropurine-2,6-dione; [0263]
1-(2-methylpropyl)-3-methyl-8-{1-[(3-fluorophenyl)methyl]pyrazol-4-yl)-1,-
3,7-trihydropurine-2,6-dione; [0264]
3-ethyl-1-(2-methylpropyl)-8-(1-{[6-(trifluoromethyl)(3-pyridyl)]methyl}p-
yrazol-4-yl)-1,3,7-trihydropurine-2,6-dione; [0265]
1-ethyl-3-methyl-8-{1-[(3-fluorophenyl)methyl]pyrazol-4-yl}-1,3,7-trihydr-
opurine-2,6-dione; and [0266]
3-ethyl-1-propyl-8-[1-(2-pyridylmethyl)pyrazol-4-yl]-1,3,7-trihydropurine-
-2,6-dione.
C. Preparation of a Compound of Formula I, Varying R.sup.1,
R.sup.2, X, Y, and Z
[0267] Similarly, following the procedure of Example 8A, but
replacing
N-(6-amino-1-ethyl-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[3-(-
trifluoromethyl)phenyl]-methyl)pyrazol-3-yl)carboxamide with other
compounds of formula (3), other compounds of Formula I are
prepared.
EXAMPLE 9
Preparation of a Compound of Formula (10)
[0268] A. Preparation of a Compound of Formula (10) in which
R.sup.1 is n-Propyl
##STR00056##
[0269] A mixture of 6-aminouracil (5.08 g, 40 mmol),
hexamethyldisilazane (50 ml), and ammonium sulfate (260 mg, 1.96
mmol) was refluxed for 12 hours. After cooling, the solid was
filtered off, and solvent was removed from the filtrate under
reduced pressure to provide the trimethylsilylated derivative of
6-aminouracil.
[0270] The product was dissolved in toluene (1.5 ml), and
iodopropane (7.8 ml, 80 mmol) and heated in an oil bath at
120.degree. C. for 2 hours. The reaction mixture was then cooled to
0.degree. C., and saturated aqueous sodium bicarbonate added
slowly. The resulting precipitate was filtered off, and washed
sequentially with water, toluene, and ether, to provide
6-amino-3-propyl-1,3-dihydropyrimidine-2,4-dione, a compound of
formula (10), which was used in the next reaction with no further
purification.
[0271] .sup.1H-NMR (DMSO-d6) .delta. 10.34 (s, 1H), 6.16 (s, 2H),
4.54 (s, 1H), 3.57-3.62 (m, 2H), 1.41-1.51 (m, 2H), 0.80 (t, 3H,
J=7.43 Hz).
B. Preparation of a Compound of Formula (10), Varying R.sup.1
[0272] Similarly, following the procedure of Example 9A, but
replacing iodopropane with other alkyl halides of formula
R.sup.1Hal, other compounds of formula (10) are prepared,
including: [0273]
6-amino-3-cyclopropylmethyl-1,3-dihydropyrimidine-2,4-dione; and
[0274]
6-amino-3-(2-methylpropyl)-1,3-dihydropyrimidine-2,4-dione.
EXAMPLE 10
Preparation of a Compound of Formula (11)
[0275] A. Preparation of a Compound of Formula (10) in which
R.sup.1 is n-Propyl
##STR00057##
[0276] To a solution of
6-amino-3-propyl-1,3-dihydropyrimidine-2,4-dione (5.6 g) in a
mixture of 50% acetic acid/water (160 ml) at 70.degree. C. was
added sodium nitrite (4.5 g) in portions over a period of 15
minutes. The mixture was stirred at 70.degree. C. for 45 minutes,
then the reaction mixture concentrated to a low volume under
reduced pressure. The solid was filtered off, and washed with
water, to provide
6-amino-5-nitroso-3-propyl-1,3-dihydropyrimidine-2,4-dione, a
compound of formula (11).
[0277] .sup.1H-NMR (DMSO-d6) .delta. 11.42 (s, 1H), 7.98 (s, 1H),
3.77-3.81 (m, 2H), 3.33 (s, 1H), 1.55-1.64 (m, 2H), 0.89 (t, 3H,
J=7.43 Hz); MS m/z 198.78 (M.sup.+), 220.78 (M.sup.++Na)
B. Preparation of a Compound of Formula (11), Varying R.sup.1
[0278] Similarly, following the procedure of Example 10A, but
replacing 6-amino-3-propyl-1,3-dihydropyrimidine-2,4-dione with
other compounds of formula (10), other compounds of formula (11)
are prepared, including: [0279]
6-amino-5-nitroso-3-cyclopropylmethyl-1,3-dihydropyrimidine-2,4-di-
one; and [0280]
6-amino-5-nitroso-3-(2-methylpropyl)-1,3-dihydropyrimidine-2,4-dione.
EXAMPLE 11
Preparation of a Compound of Formula (12)
[0281] A. Preparation of a Compound of Formula (12) in which R' is
n-Propyl
##STR00058##
[0282] To a solution of
6-amino-5-nitroso-3-propyl-1,3-dihydropyrimidine-2,4-dione (5.4 g,
27 mmol) in 12.5% aqueous ammonia (135 ml) at 70.degree. C. was
added sodium dithionite (Na.sub.2S.sub.2O.sub.4, 9.45 g, 54 mmol)
in portions over 15 minutes, and the mixture was stirred for 20
minutes. The solution was concentrated under reduced pressure,
cooled to 5.degree. C., the precipitate filtered off, and washed
with cold water, to provide
5,6-diamino-3-propyl-1,3-dihydropyrimidine-2,4-dione, a compound of
formula (12).
[0283] .sup.1H-NMR (DMSO-d6) .delta. 0.81 (t, 3H, J=7.43 Hz),
1.43-1.52 (m, 2H), 3.63-3.67 (m, 2H), 5.56 (s, 2H); MS m/z 184.95
(M.sup.+), 206.96 (M.sup.++Na)
B. Preparation of a Compound of Formula (12), Varying R.sup.1
[0284] Similarly, following the procedure of Example 11A, but
replacing 6-amino-3-propyl-1,3-dihydropyrimidine-2,4-dione with
other compounds of formula (11), other compounds of formula (12)
are prepared, including: [0285]
5,6-diamino-3-cyclopropylmethyl-1,3-dihydropyrimidine-2,4-dione;
and [0286]
5,6-diamino-3-(2-methylpropyl)-1,3-dihydropyrimidine-2,4-dione.
EXAMPLE 12
Preparation of a Compound of Formula (13)
[0287] A. Preparation of a Compound of Formula (13) in which
R.sup.1 is n-Propyl, X is 1,4-Pyrazolyl, Y is Methylene, and Z is
3-Trifluoromethylphenyl
##STR00059##
[0288] To a mixture of
5,6-diamino-3-propyl-1,3-dihydropyrimidine-2,4-dione (2.3 g, 126
mmol) and
1-{[3-(trifluoromethyl)phenyl]methyl}pyrazole-4-carboxylic acid
(3.79 g, 14 mmol) in methanol (50 ml) was added
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.67
g, 14 mmol), and the reaction mixture was stirred for 3 days at
room temperature (although less time is acceptable). The
precipitate was filtered off, and was washed sequentially with
water, and methanol. The product was dried under vacuum to provide
N-(6-amino-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[3-(trifluor-
omethyl)phenyl]methyl}pyrazol-4-yl)carboxamide, a compound of
formula (13).
[0289] .sup.1H-NMR (DMSO-d6) .delta. 10.44 (s, 1H), 8.56 (s, 1H),
8.37 (s, 1H), 8.00 (s, 1H), 7.56-7.71 (m, 3H), 6.02 (s, 1H), 5.49
(s, 2H), 3.62-3.66 (m, 2H), 1.44-1.53 (m, 2H), 0.82 (t, 3H, J=7.43
Hz); MS m/z 458.92 (M.sup.++Na).
B. Alternative Preparation of a Compound of Formula (3) in which
R.sup.1 is n-Propyl, X is 1,4-Pyrazolyl, Y is Methylene, and Z is
3-Trifluoromethylphenyl
[0290] A solution of
1-{[3-(trifluoromethyl)phenyl]methyl}pyrazole-4-carboxylic acid (1
g, 3.7 mmol) in thionyl chloride (1 ml) was heated at 70.degree. C.
for 4 hours. Excess thionyl chloride was distilled off, and the
residue treated with methylene chloride/hexanes. The solvent was
removed under reduced pressure, and the residue dissolved in
acetonitrile. This solution was added to a suspension of
5,6-diamino-3-propyl-1,3-dihydropyrimidine-2,4-dione (2.3 g, 126
mmol) and triethylamine (1 ml) in acetonitrile (20 ml) at 0.degree.
C., and stirred for 16 hours. The reaction mixture was quenched
with water (5 ml), acidified with hydrochloric acid, stirred for 30
minutes, and the precipitate filtered off. The product was washed
with ether, to provide
N-(6-amino-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[3-(trifluor-
omethyl)phenyl]methyl}pyrazol-4-yl)carboxamide, a compound of
formula (13).
C. Preparation of a Compound of Formula (13), Varying R.sup.1, X,
Y, and Z
[0291] Similarly, following the procedure of Example 12A or 12B,
but optionally replacing
6-amino-3-propyl-1,3-dihydropyrimidine-2,4-dione with other
compounds of formula (12), and optionally replacing
1-{[3-(trifluoromethyl)phenyl]methyl}pyrazole-4-carboxylic acid
with other compounds of formula Z-Y--X--CO.sub.2H, other compounds
of formula (13) are prepared, including: [0292]
N-(6-amino-2,4-dioxo-3-cyclopropylmethyl(1,3-dihydropyrimidin-5-yl))(1-{[-
3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)carboxamide; [0293]
N-(6-amino-2,4-dioxo-3-(2-methylpropyl)(1,3-dihydropyrimidin-5-yl))(1-{[3-
-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)carboxamide; [0294]
N-(6-amino-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[3-fluorophe-
nyl]methyl}pyrazol-4-yl)carboxamide; [0295]
N-(6-amino-2,4-dioxo-3-cyclopropylmethyl(1,3-dihydropyrimidin-5-yl))(1-{[-
3-fluorophenyl]methyl}pyrazol-4-yl)carboxamide; [0296]
N-(6-amino-2,4-dioxo-3-(2-methylpropyl)(1,3-dihydropyrimidin-5-yl))(1-{[3-
-fluorophenyl]methyl}pyrazol-4-yl)carboxamide; [0297]
N-(6-amino-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-[1-benzyl]pyr-
azol-4-yl)carboxamide; [0298]
N-(6-amino-2,4-dioxo-3-cyclopropylmethyl(1,3-dihydropyrimidin-5-yl))(1-[1-
-benzyl]pyrazol-4-yl)carboxamide; [0299]
N-(6-amino-2,4-dioxo-3-(2-methylpropyl)(1,3-dihydropyrimidin-5-yl))(1-[1--
benzyl]pyrazol-4-yl)carboxamide; [0300]
N-(6-amino-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[3-cyanophen-
yl]methyl}pyrazol-4-yl)carboxamide; [0301]
N-(6-amino-2,4-dioxo-3-cyclopropylmethyl(1,3-dihydropyrimidin-5-yl))(1-{[-
3-cyanophenyl]methyl}pyrazol-4-yl carboxamide; [0302]
N-(6-amino-2,4-dioxo-3-(2-methylpropyl)(1,3-dihydropyrimidin-5-yl))(1-{[3-
-cyanophenyl]methyl}pyrazol-4-yl)carboxamide; [0303]
N-(6-amino-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[1-(2-(1H-1,-
2,3,4-tetraazol-5-yl)ethyl)pyrazol-4-yl}carboxamide; [0304]
N-(6-amino-2,4-dioxo-3-cyclopropylmethyl(1,3-dihydropyrimidin-5-yl))(1-{[-
-(2-(1H-1,2,3,4-tetraazol-5-yl)ethyl)pyrazol-4-yl)carboxamide;
[0305]
N-(6-amino-2,4-dioxo-3-(2-methylpropyl)(1,3-dihydropyrimidin-5-yl))(1-{[1-
-(2-(1H-1,2,3,4-tetraazol-5-yl)ethyl)pyrazol-4-yl)carboxamide;
[0306]
N-(6-amino-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[6-(trifluor-
omethyl)(3-pyridyl)]methyl}pyrazol-4-yl)carboxamide; [0307]
N-(6-amino-2,4-dioxo-3-cyclopropylmethyl(1,3-dihydropyrimidin-5-yl))(1-{[-
6-(trifluoromethyl)(3-pyridyl)]methyl}pyrazol-4-yl)carboxamide; and
[0308]
N-(6-amino-2,4-dioxo-3-(2-methylpropyl)(1,3-dihydropyrimidin-5-yl))(1-{[6-
-(trifluoromethyl)(3-pyridyl)]methyl}pyrazol-4-yl)carboxamide.
EXAMPLE 13
Preparation of a Compound of Formula (3)
[0309] A. Preparation of a Compound of Formula (3) in which R.sup.1
is n-Propyl, R.sup.2 is Ethyl, X is 1,4-Pyrazolyl, Y is Methylene,
and Z is 3-Trifluoromethylphenyl
##STR00060##
[0310] A mixture of a solution of
N-(6-amino-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[3-(trifluor-
omethyl)-phenyl]methyl}pyrazol-3-yl)carboxamide (872 mg, 2 mmol) in
dimethylformamide (10 ml), potassium carbonate (552 mg, 4 mmol) and
ethyl iodide (0.24 ml, 3 mmol) was stirred at room temperature
overnight. The reaction mixture was filtered, and the solvent was
evaporated from the filtrate under reduced pressure. The residue
was stirred with water for two hours at room temperature, and the
precipitate filtered off, washed with water, and then dissolved in
methanol. The solvent was then removed under reduced pressure to
provide
N-(6-amino-1-ethyl-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[3-(-
trifluoromethyl)phenyl]methyl}pyrazol-4-yl)carboxamide, a compound
of formula (3).
[0311] .sup.1H-NMR (DMSO-d6): .delta. 8.58 (s, 1H), 8.39 (s, 1H),
8.01 (s, 1H), 7.72-7.50 (m, 4H), 6.71 (s, 2H), 5.51 (s, 2H),
4.0-3.82 (m, 2H), 3.77-3.65 (m, 2H), 1.60-1.50 (m, 2H), 1.13 (t,
3H, J=6.8 Hz), 0.84 (t, 3H, J=7.2 Hz); MS m/z 462.9 (M.sup.-)
B. Preparation of a Compound of Formula (13), Varying R.sup.1, X,
Y, and Z
[0312] Similarly, following the procedure of Example 13A, but
replacing
N-(6-amino-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[3-(trifluor-
omethyl)-phenyl]methyl}pyrazol-3-yl)carboxamide with other
compounds of formula (13), other compounds of formula (3) are
prepared, including: [0313]
N-(6-amino-1-methyl-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))-
(1-{[3-(trifluoromethyl)phenyl]methyl}-pyrazol-4-yl)carboxamide;
[0314]
N-(6-amino-1-methyl-2,4-dioxo-3-cyclopropylmethyl(1,3-dihydropyrimidin-5--
yl))(1-{[3-(trifluoromethyl)phenyl]methyl}-pyrazol-4-yl)carboxamide;
[0315]
N-(6-amino-1-ethyl-2,4-dioxo-3-cyclopropylmethyl(1,3-dihydropyrimi-
din-5-yl))(1-{[3-(trifluoromethyl)phenyl]methyl}-pyrazol-4-yl)carboxamide;
[0316]
N-(6-amino-1-methyl-2,4-dioxo-3-ethyl(1,3-dihydropyrimidin-5-yl))(-
1-{[3-fluorophenyl]methyl}-pyrazol-4-yl)carboxamide; [0317]
N-(6-amino-1-methyl-2,4-dioxo-3-cyclopropylmethyl(1,3-dihydropyrimidin-5--
yl))(1-{[3-fluorophenyl]methyl}-pyrazol-4-yl)carboxamide; [0318]
N-(6-amino-1-ethyl-2,4-dioxo-3-cyclopropylmethyl(1,3-dihydropyrimidin-5-y-
l))(1-{[3-fluorophenyl]methyl}-pyrazol-4-yl)carboxamide; [0319]
N-[6-amino-3-(cyclopropylmethyl)-1-methyl-2,4-dioxo(1,3-dihydropyrimidin--
5-yl)][1-benzylpyrazol-4-yl]carboxamide; [0320]
N-(6-amino-1-methyl-2,4-dioxo-3-cyclopropylmethyl(1,3-dihydropyrimidin-5--
yl))(1-{[3-cyanophenyl]methyl}-pyrazol-4-yl)carboxamide; [0321]
[1-(2-(1H-1,2,3,4-tetraazol-5-yl)ethyl)pyrazol-4-yl]-N-[6-amino-3-(cyclop-
ropylmethyl)-1-methyl-2,4-dioxo(1,3-dihydropyrimidin-5-yl)]carboxamide;
[0322]
N-[6-amino-3-(cyclopropylmethyl)-1-ethyl-2,4-dioxo(1,3-dihydropyri-
midin-5-yl)](1-{[6-(trifluoromethyl)(3-pyridyl)]methyl}pyrazol-4-yl)carbox-
amide; [0323]
N-[6-amino-3-propyl)-1-ethyl-2,4-dioxo(1,3-dihydropyrimidin-5-yl)](1-{(2--
pyridyl)]methyl}pyrazol-4-yl)carboxamide; [0324]
N-[6-amino-3-(2-methylpropyl)-1-methyl-2,4-dioxo(1,3-dihydropyrimidin-5-y-
l)][1-benzylpyrazol-4-yl]carboxamide; [0325]
N-[6-amino-3-(2-methylpropyl)-1-methyl-2,4-dioxo(1,3-dihydropyrimidin-5-y-
l)][1-{[3-fluorophenyl]methyl}pyrazol-4-yl]carboxamide; [0326]
N-[6-amino-3-(2-methylpropyl)-1-ethyl-2,4-dioxo(1,3-dihydropyrimidin-5-yl-
)][1-{[3-fluorophenyl]methyl}pyrazol-4-yl]carboxamide; [0327]
N-[6-amino-3-(2-methylpropyl)-1-methyl-2,4-dioxo(1,3-dihydropyrimidin-5-y-
l)][1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl]carboxamide;
and [0328]
N-[6-amino-3-(2-methylpropyl)-1-ethyl-2,4-dioxo(1,3-dihydropyrimid-
in-5-yl)](1-{[6-(trifluoromethyl)(3-pyridyl)]methyl}pyrazol-4-yl)carboxami-
de.
EXAMPLE 14
Preparation of a Compound of Formula I
[0329] A. Preparation of a Compound of Formula I in which R.sup.1
is n-Propyl, R.sup.2 is Ethyl, X is 1,4-Pyrazolyl, Y is Methylene,
and Z is 3-Trifluoromethylphenyl
##STR00061##
[0330] A mixture of
N-(6-amino-1-ethyl-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[3-(-
trifluoromethyl)phenyl]methyl}pyrazol-3-yl)carboxamide (850 mg,
2.34 mmol), 10% aqueous sodium hydroxide (10 ml), and methanol (10
ml) was stirred at 100.degree. C. for 18 hours. The mixture was
cooled, methanol removed under reduced pressure, and the remaining
mixture was acidified with hydrochloric acid to pH 2. The
precipitate was filtered off, washed with water/methanol mixture,
to provide
3-ethyl-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,-
3,7-trihydropurine-2,6-dione, a compound of Formula I.
[0331] .sup.1H-NMR (DMSO-d6) .delta. 8.57 (s, 1H), 8.15 (s, 1H),
7.60-7.75 (m, 4H), 5.54 (s, 2H), 4.05-4.50 (m, 2H), 3.87-3.91 (m,
2H), 1.55-1.64 (m, 2H), 1.25 (t, 3H, J=7.03 Hz), 0.90 (t, 3H,
J=7.42 Hz); MS m/z 447.2 (M.sup.+)
B. Preparation of a Compound of Formula I, Varying R.sup.1,
R.sup.2, X, Y, and Z
[0332] Similarly, following the procedure of Example 14A, but
replacing
N-(6-amino-1-ethyl-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[3-(-
trifluoromethyl)phenyl]methyl}pyrazol-3-yl)carboxamide with other
compounds of formula (13), other compounds of Formula I are
prepared, including those listed in Example 8.
EXAMPLE 15
Preparation of a Compound of Formula (14)
[0333] A. Preparation of a Compound of Formula (14) in which
R.sup.2 is Ethyl
##STR00062##
[0334] To a solution of
6-amino-1-ethyl-1,3-dihydropyrimidine-2,4-dione (5.0 g, 32.3 mmol)
in a mixture of 50% acetic acid/water (50 ml) at 70.degree. C. was
added sodium nitrite (4.45 g, 64.5 mmol) in portions over a period
of 30 minutes. The mixture was stirred at 70.degree. C. for a
further 30 minutes. The reaction mixture was cooled, and the
precipitate filtered off, and washed with water, then methanol, to
provide 6-amino-1-ethyl-5-nitroso-1,3-dihydropyrimidine-2,4-dione,
a compound of formula (14).
[0335] .sup.1H-NMR (DMSO-d6): .delta. 11.52 (s, 1H), 9.16 (s, 1H),
3.83 (q, 2H, J=7.0 Hz), 1.11 (t, 3H, J=7.0 Hz). MS m/z 184.8
(M.sup.+), 206-80 (M.sup.++Na)
B. Preparation of a Compound of Formula (14), Varying R.sup.2
[0336] Similarly, following the procedure of Example 15A, but
replacing 6-amino-1-ethyl-1,3-dihydropyrimidine-2,4-dione with
6-amino-1-methyl-1,3-dihydropyrimidine-2,4-dione,
6-amino-1-methyl-5-nitroso-1,3-dihydropyrimidine-2,4-dione was
prepared.
C. Preparation of a Compound of Formula (14), Varying R.sup.2
[0337] Similarly, following the procedure of Example 15A, but
replacing 6-amino-1-ethyl-1,3-dihydropyrimidine-2,4-dione with
other compounds of formula (5), other compounds of formula (14) are
prepared.
EXAMPLE 16
Preparation of a Compound of Formula (15)
[0338] A. Preparation of a Compound of Formula (15) in which
R.sup.2 is Ethyl
##STR00063##
[0339] To a solution of
6-amino-1-ethyl-5-nitroso-1,3-dihydropyrimidine-2,4-dione (3.9 g,
21.2 mmol) in 14.5% aqueous ammonia (50 ml) at 50.degree. C. was
added sodium dithionite (Na.sub.2S.sub.2O.sub.4, 7.37 g, 42.4 mmol)
in portions over 15 minutes, and the mixture was stirred for 20
minutes. The solution was concentrated under reduced pressure to a
volume of 30 ml, cooled to 5.degree. C., the precipitate filtered
off, and washed with cold water, to provide
5,6-diamino-1-ethyl-1,3-dihydropyrimidine-2,4-dione, a compound of
formula (15).
[0340] .sup.1H-NMR (DMSO-d6): .delta. 10.58 (s, 1H), 6.18 (s, 2H),
3.83 (q, 2H, J=7.2 Hz), 2.82 (s, 2H), 1.10 (t, 3H, J=7.2 Hz).
B. Preparation of a Compound of Formula (15), Varying R.sup.2
[0341] Similarly, following the procedure of Example 16A, but
replacing 6-amino-1-ethyl-5-nitroso-1,3-dihydropyrimidine-2,4-dione
with 6-amino-1-methyl-5-nitroso-1,3-dihydropyrimidine-2,4-dione,
5,6-diamino-1-methyl-1,3-dihydropyrimidine-2,4-dione was
prepared.
C. Preparation of a Compound of Formula (15), Varying R.sup.2
[0342] Similarly, following the procedure of Example 16A, but
replacing 6-amino-1-ethyl-5-nitroso-1,3-dihydropyrimidine-2,4-dione
with other compounds of formula (14), other compounds of formula
(15) are prepared.
EXAMPLE 17
Preparation of a Compound of Formula (16)
[0343] A. Preparation of a Compound of Formula (16) in which
R.sup.2 is Ethyl, X is 1,4-Pyrazolyl, Y is Methylene, and Z is
3-Trifluoromethylphenyl
##STR00064##
[0344] To a mixture of
5,6-diamino-1-ethyl-1,3-dihydropyrimidine-2,4-dione (2 g, 11.76
mmol) and
1-{[3-(trifluoromethyl)phenyl]methyl}pyrazole-4-carboxylic acid
(3.5 g, 12.94 mmol) in methanol (50 ml) was added
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.47
g, 12.94 mmol), and the reaction mixture was stirred for 16 hours
at room temperature. Solvent was removed under reduced pressure,
and the residue was washed with water and methanol. The product was
dried under vacuum to provide
N-(6-amino-1-ethyl-2,4-dioxo(1,3-dihydropyrimidin-5-yl))(1-{[3-(t-
rifluoromethyl)phenyl]methyl}pyrazol-4-yl)carboxamide, a compound
of formula (16).
[0345] .sup.1H-NMR (DMSO-d6): .delta. 10.60 (s, 1H), 8.50 (s, 1H),
8.39 (s, 1H), 8.01 (s, 1H), 7.72-7.50 (m, 4H), 6.69 (s, 2H), 5.50
(s, 2H), 3.87 (q, 2H, J=7.2 Hz), 1.11 (t, 3H, 7.2 Hz); MS m/z 421
(M.sup.-)
B. Preparation of a Compound of Formula (16), Varying R.sup.2, X,
Y, and Z
[0346] Similarly, following the procedure of Example 17A, but
replacing 5,6-diamino-1-ethyl-1,3-dihydropyrimidine-2,4-dione with
5,6-diamino-1-methyl-1,3-dihydropyrimidine-2,4-dione,
N-(6-amino-1-methyl-2,4-dioxo(1,3-dihydropyrimidin-5-yl))(1-{[3-(trifluor-
omethyl)phenyl]methyl}pyrazol-4-yl)carboxamide was prepared.
C. Preparation of a Compound of Formula (16), Varying R.sup.2, X,
Y, and Z
[0347] Similarly, following the procedure of Example 16A, but
replacing 5,6-diamino-1-ethyl-1,3-dihydropyrimidine-2,4-dione with
other compounds of formula (14), other compounds of formula (15)
are prepared.
EXAMPLE 18
Preparation of a Compound of Formula (3)
[0348] A. Preparation of a Compound of Formula (3) in which R.sup.1
is n-Propyl, R.sup.2 is Ethyl, X is 1,4-Pyrazolyl, Y is Methylene,
and Z is 3-Trifluoromethylphenyl
##STR00065##
[0349] A mixture of a solution of
N-(6-amino-1-ethyl-2,4-dioxo(1,3-dihydropyrimidin-5-yl))(1-{[3-(trifluoro-
methyl)phenyl]methyl}pyrazol-3-yl)carboxamide (1.5 g, 3.55 mmol) in
dimethylformamide (30 ml), potassium carbonate (980 mg, 7.1 mmol)
and propyl iodide (724 mg, 4.26 mmol) was stirred at room
temperature overnight. Water was added, and the precipitate
filtered off, to provide
N-(6-amino-1-ethyl-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[3-(-
trifluoromethyl)phenyl]methyl}pyrazol-4-yl)carboxamide, a compound
of formula (3), which was used in the next reaction with no further
purification.
[0350] .sup.1H-NMR (DMSO-d6): .delta. 8.58 (s, 1H), 8.39 (s, 1H),
8.01 (s, 4H), 7.72-7.50 (m, 4H), 6.71 (s, 2H), 5.51 (s, 2H),
4.0-3.82 (m, 2H), 3.77-3.65 (m, 2H), 1.60-1.50 (m, 2H), 1.13 (t,
3H, J=6.8 Hz), 0.84 (t, 3H, J=7.2 Hz); MS m/z 462.9 (M.sup.-)
B. Preparation of a Compound of Formula (3), Varying R.sup.1,
R.sup.2, X, Y, and Z
[0351] Similarly, following the procedure of Example 18A, but
replacing
N-(6-amino-1-ethyl-2,4-dioxo(1,3-dihydropyrimidin-5-yl))(1-{[3-(trifluoro-
methyl)phenyl]-methyl)pyrazol-3-yl)carboxamide with
N-(6-amino-1-methyl-2,4-dioxo(1,3-dihydropyrimidin-5-yl)),
N-(6-amino-1-methyl-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[3--
(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)carboxamide was
prepared.
C. Preparation of a Compound of Formula (3), Varying R.sup.1,
R.sup.2, X, Y, and Z
[0352] Similarly, following the procedure of Example 18A, but
optionally replacing
N-(6-amino-1-ethyl-2,4-dioxo(1,3-dihydropyrimidin-5-yl))(1-{[3--
(trifluoromethyl)phenyl]methyl}pyrazol-3-yl)carboxamide with other
compounds of formula (15), and optionally replacing propyl iodide
with other compounds of formula R.sup.-Hal, other compounds of
formula (3) are prepared.
EXAMPLE 19
Preparation of a Compound of Formula I
[0353] A. Preparation of a Compound of Formula I in which R.sup.1
is n-Propyl, R.sup.2 is Ethyl, X is 1,4-Pyrazolyl, Y is Methylene,
and Z is 3-Trifluoromethylphenyl
##STR00066##
[0354] A mixture of
N-(6-amino-1-ethyl-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1-{[3-(-
trifluoromethyl)phenyl]methyl}pyrazol-3-yl)carboxamide (300 mg, 464
mmol), 20% aqueous sodium hydroxide (5 ml), and methanol (10 ml)
was stirred at 80.degree. C. for 3 hours. The mixture was cooled,
methanol removed under reduced pressure, and the remaining mixture
was acidified with hydrochloric acid to pH 2. The precipitate was
filtered off, washed with water and methanol, to provide
3-ethyl-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,-
3,7-trihydropurine-2,6-dione, a compound of Formula I.
[0355] .sup.1H-NMR (DMSO-d6) .delta. 8.57 (s, 1H), 8.15 (s, 1H),
7.60-7.75 (m, 4H), 5.54 (s, 2H), 4.05-4.50 (m, 2H), 3.87-3.91 (m,
2H), 1.55-1.64 (m, 2H), 1.25 (t, 3H, J=7.03 Hz), 0.90 (t, 3H,
J=7.42 Hz); MS m/z 447.2 (M.sup.+)
EXAMPLE 20
[0356] Hard gelatin capsules containing the following ingredients
are prepared:
TABLE-US-00001 Quantity Ingredient (mg/capsule) Active Ingredient
30.0 Starch 305.0 Magnesium stearate 5.0
The above ingredients are mixed and filled into hard gelatin
capsules.
EXAMPLE 21
[0357] A tablet formula is prepared using the ingredients
below:
TABLE-US-00002 Quantity Ingredient (mg/tablet) Active Ingredient
25.0 Cellulose, microcrystalline 200.0 Colloidal silicon dioxide
10.0 Stearic acid 5.0
The components are blended and compressed to form tablets.
EXAMPLE 22
[0358] A dry powder inhaler formulation is prepared containing the
following components:
TABLE-US-00003 Ingredient Weight % Active Ingredient 5 Lactose
95
The active ingredient is mixed with the lactose and the mixture is
added to a dry powder inhaling appliance.
EXAMPLE 23
[0359] Tablets, each containing 30 mg of active ingredient, are
prepared as follows:
TABLE-US-00004 Quantity Ingredient (mg/tablet) Active Ingredient
30.0 mg Starch 45.0 mg Microcrystalline cellulose 35.0 mg
Polyvinylpyrrolidone 4.0 mg (as 10% solution in sterile water)
Sodium carboxymethyl starch 4.5 mg Magnesium stearate 0.5 mg Talc
1.0 mg Total 120 mg
[0360] The active ingredient, starch and cellulose are passed
through a No. 20 mesh U.S. sieve and mixed thoroughly. The solution
of polyvinylpyrrolidone is mixed with the resultant powders, which
are then passed through a 16 mesh U.S. sieve. The granules so
produced are dried at 50.degree. C. to 60.degree. C. and passed
through a 16 mesh U.S. sieve. The sodium carboxymethyl starch,
magnesium stearate, and talc, previously passed through a No. 30
mesh U.S. sieve, are then added to the granules which, after
mixing, are compressed on a tablet machine to yield tablets each
weighing 120 mg.
EXAMPLE 24
[0361] Suppositories, each containing 25 mg of active ingredient
are made as follows:
TABLE-US-00005 Ingredient Amount Active Ingredient 25 mg Saturated
fatty acid glycerides to 2,000 mg
[0362] The active ingredient is passed through a No. 60 mesh U.S.
sieve and suspended in the saturated fatty acid glycerides
previously melted using the minimum heat necessary. The mixture is
then poured into a suppository mold of nominal 2.0 g capacity and
allowed to cool.
EXAMPLE 25
[0363] Suspensions, each containing 50 mg of active ingredient per
5.0 mL dose are made as follows:
TABLE-US-00006 Ingredient Amount Active Ingredient 50.0 mg Xanthan
gum 4.0 mg Sodium carboxymethyl cellulose (11%) Microcrystalline
cellulose (89%) 50.0 mg Sucrose 1.75 g Sodium benzoate 10.0 mg
Flavor and Color q.v. Purified water to 5.0 mL
[0364] The active ingredient, sucrose and xanthan gum are blended,
passed through a No. 10 mesh U.S. sieve, and then mixed with a
previously made solution of the microcrystalline cellulose and
sodium carboxymethyl cellulose in water. The sodium benzoate,
flavor, and color are diluted with some of the water and added with
stirring. Sufficient water is then added to produce the required
volume.
EXAMPLE 26
[0365] A subcutaneous formulation may be prepared as follows:
TABLE-US-00007 Ingredient Quantity Active Ingredient 5.0 mg Corn
Oil 1.0 mL
EXAMPLE 27
[0366] An injectable preparation is prepared having the following
composition:
TABLE-US-00008 Ingredients Amount Active ingredient 2.0 mg/ml
Mannitol, USP 50 mg/ml Gluconic acid, USP q.s. (pH 5-6) water
(distilled, sterile) q.s. to 1.0 ml Nitrogen Gas, NF q.s.
EXAMPLE 28
[0367] A topical preparation is prepared having the following
composition:
TABLE-US-00009 Ingredients grams Active ingredient 0.2-10 Span 60
2.0 Tween 60 2.0 Mineral oil 5.0 Petrolatum 0.10 Methyl paraben
0.15 Propyl paraben 0.05 BHA (butylated hydroxy anisole) 0.01 Water
q.s. to 100
[0368] All of the above ingredients, except water, are combined and
heated to 60.degree. C. with stirring. A sufficient quantity of
water at 60.degree. C. is then added with vigorous stirring to
emulsify the ingredients, and water then added q.s. 100 g.
EXAMPLE 29
Sustained Release Composition
TABLE-US-00010 [0369] Weight Preferred Most Ingredient Range (%)
Range (%) Preferred Active ingredient 50-95 70-90 75
Microcrystalline cellulose (filler) 1-35 5-15 10.6 Methacrylic acid
copolymer 1-35 5-12.5 10.0 Sodium hydroxide 0.1-1.0 0.2-0.6 0.4
Hydroxypropyl methylcellulose 0.5-5.0 1-3 2.0 Magnesium stearate
0.5-5.0 1-3 2.0
[0370] The sustained release formulations of this invention are
prepared as follows: compound and pH-dependent binder and any
optional excipients are intimately mixed (dry-blended). The
dry-blended mixture is then granulated in the presence of an
aqueous solution of a strong base which is sprayed into the blended
powder. The granulate is dried, screened, mixed with optional
lubricants (such as talc or magnesium stearate), and compressed
into tablets. Preferred aqueous solutions of strong bases are
solutions of alkali metal hydroxides, such as sodium or potassium
hydroxide, preferably sodium hydroxide, in water (optionally
containing up to 25% of water-miscible solvents such as lower
alcohols).
[0371] The resulting tablets may be coated with an optional
film-forming agent, for identification, taste-masking purposes and
to improve ease of swallowing. The film forming agent will
typically be present in an amount ranging from between 2% and 4% of
the tablet weight. Suitable film-forming agents are well known to
the art and include hydroxypropyl, methylcellulose, cationic
methacrylate copolymers (dimethylaminoethyl
methacrylate/methyl-butyl methacrylate copolymers--Eudragit.RTM.
E--Rohm. Pharma), and the like. These film-forming agents may
optionally contain colorants, plasticizers, and other supplemental
ingredients.
[0372] The compressed tablets preferably have a hardness sufficient
to withstand 8 Kp compression. The tablet size will depend
primarily upon the amount of compound in the tablet. The tablets
will include from 300 to 1100 mg of compound free base. Preferably,
the tablets will include amounts of compound free base ranging from
400-600 mg, 650-850 mg, and 900-1100 mg.
[0373] In order to influence the dissolution rate, the time during
which the compound containing powder is wet mixed is controlled.
Preferably the total powder mix time, i.e. the time during which
the powder is exposed to sodium hydroxide solution, will range from
1 to 10 minutes and preferably from 2 to 5 minutes. Following
granulation, the particles are removed from the granulator and
placed in a fluid bed dryer for drying at about 60.degree. C.
EXAMPLE 30
A.sub.2B Adenosine Receptor Assays
Methods
Radioligand Binding for A.sub.2B Adenosine Receptor.
[0374] Human A.sub.2B adenosine receptor cDNA was stably
transfected into HEK-293 cells (referred to as HEK-A2B cells).
Monolayer of HEK-A2B cells were washed with PBS once and harvested
in a buffer containing 10 mM HEPES (pH 7.4), 10 mM EDTA and
protease inhibitors. These cells were homogenized in polytron for 1
minute at setting 4 and centrifuged at 29000 g for 15 minutes at
4.degree. C. The cell pellets were washed once with a buffer
containing 10 mM HEPES (pH7.4), 1 mM EDTA and protease inhibitors,
and were resuspended in the same buffer supplemented with 10%
sucrose. Frozen aliquots were kept at -80.degree. C. Competition
assays were started by mixing 10 nM .sup.3H-ZM241385 (Tocris
Cookson) with various concentrations of test compounds and 50 .mu.g
membrane proteins in TE buffer (50 mM Tris and 1 mM EDTA)
supplemented with 1 Unit/mL adenosine deaminase. The assays were
incubated for 90 minutes, stopped by filtration using Packard
Harvester and washed four times with ice-cold TM buffer (10 mM
Tris, 1 mM MgCl2, pH 7.4). Non specific binding was determined in
the presence of 10 .mu.M ZM241385. The affinities of compounds
(i.e. Ki values) were calculated using GraphPad software.
Radioligand Binding for Other Adenosine Receptors.
[0375] Human A.sub.1, A.sub.2A, A.sub.3 adenosine receptor cDNAs
were stably transfected into either CHO or HEK-293 cells (referred
to as CHO-A1, HEK-A2A, CHO-A3). Membranes were prepared from these
cells using the same protocol as described above. Competition
assays were started by mixing 0.5 nM .sup.3H-CPX (for CHO-A1), 2 nM
.sup.3H-ZM241385 (HEK-A2A) or 0.1 nM .sup.125I-AB-MECA (CHO-A3)
with various concentrations of test compounds and the perspective
membranes in TE buffer (50 mM Tris and 1 mM EDTA fo CHO-A1 and
HEK-A2A) or TEM buffer (50 mM Tris, 1 mM EDTA and 10 mM MgCl.sub.2
for CHO-A3) supplemented with 1 Unit/mL adenosine deaminase. The
assays were incubated for 90 minutes, stopped by filtration using
Packard Harvester and washed four times with ice-cold TM buffer (10
mM Tris, 1 mM MgCl2, pH 7.4). Non specific binding was determined
in the presence of 1 .mu.M CPX (CHO-A1), 1 .mu.M ZM214385 (HEK-A2A)
and 1 .mu.M IB-MECA (CHO-A3). The affinities of compounds (i.e. Ki
values) were calculated using GraphPad software.
cAMP Measurements.
[0376] Monolayer of transfected cells were collected in PBS
containing 5 mM EDTA. Cells were washed once with DMEM and
resuspended in DMEM containing 1 Unit/mL adenosine deaminase at a
density of 100,000-500,000 cells/ml. 100 .mu.l of the cell
suspension was mixed with 25 .mu.l containing various agonists
and/or antagonists and the reaction was kept at 37.degree. C. for
15 minutes. At the end of 15 minutes, 125 .mu.l 0.2N HCl was added
to stop the reaction. Cells were centrifuged for 10 minutes at 1000
rpm. 100 .mu.l of the supernatant was removed and acetylated. The
concentrations of cAMP in the supernatants were measured using the
direct cAMP assay from Assay Design.
[0377] A.sub.2A and A.sub.2B adenosine receptors are coupled to Gs
proteins and thus agonists for A.sub.2A adenosine receptor (such as
CGS21680) or for A.sub.2B adenosine receptor (such as NECA)
increase the cAMP accumulations whereas the antagonists to these
receptors prevent the increase in cAMP accumulations-induced by the
agonists. A.sub.1 and A.sub.3 adenosine receptors are coupled to Gi
proteins and thus agonists for A.sub.1 adenosine receptor (such as
CPA) or for A.sub.3 adenosine receptor (such as IB-MECA) inhibit
the increase in cAMP accumulations-induced by forskolin.
Antagonists to A.sub.1 and A.sub.3 receptors prevent the inhibition
in cAMP accumulations.
[0378] The compounds of the invention were shown to be
A.sub.2B-antagonists by the above tests.
[0379] Compounds of the invention were also tested in a mouse model
for asthma, using the procedures disclosed in U.S. Pat. No.
6,387,913, the relevant portion of which is hereby incorporated by
reference, and shown to be efficacious.
* * * * *