U.S. patent application number 12/912367 was filed with the patent office on 2011-04-28 for substituted dihydroquinazolines as platelet adp receptor inhibitors.
This patent application is currently assigned to Portola Pharmaceuticals, Inc.. Invention is credited to Shawn M. Bauer, Anjali Pandey, Carrol Anna Crew Scarborough, Robert M. Scarborough.
Application Number | 20110098247 12/912367 |
Document ID | / |
Family ID | 39048759 |
Filed Date | 2011-04-28 |
United States Patent
Application |
20110098247 |
Kind Code |
A1 |
Scarborough; Robert M. ; et
al. |
April 28, 2011 |
Substituted Dihydroquinazolines as Platelet ADP Receptor
Inhibitors
Abstract
Compounds of formula I are provided: ##STR00001## wherein
Ar.sup.1 and Ar.sup.2 are substituted aromatic rings and L.sup.1
and L.sup.2 are independent divalent linking groups. The compounds
are useful as platelet ADP receptor inhibitors, for treating
thrombosis and for reducing the likelihood and/or severity of a
secondary ischemic event in a patient.
Inventors: |
Scarborough; Robert M.;
(Half Moon Bay, CA) ; Bauer; Shawn M.; (Pacifica,
CA) ; Pandey; Anjali; (Fremont, CA) ;
Scarborough; Carrol Anna Crew; (Half Moon Bay, CA) |
Assignee: |
Portola Pharmaceuticals,
Inc.
South San Francisco
CA
|
Family ID: |
39048759 |
Appl. No.: |
12/912367 |
Filed: |
October 26, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
11856616 |
Sep 17, 2007 |
7834023 |
|
|
12912367 |
|
|
|
|
60846328 |
Sep 20, 2006 |
|
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|
Current U.S.
Class: |
514/56 ;
514/230.5; 514/266.24; 514/309 |
Current CPC
Class: |
C07D 413/12 20130101;
C07D 409/12 20130101; C07D 239/96 20130101; C07D 417/14 20130101;
C07D 403/12 20130101; A61P 7/02 20180101; C07D 413/14 20130101;
C07D 409/14 20130101; A61P 43/00 20180101; C07D 265/22 20130101;
A61P 9/10 20180101 |
Class at
Publication: |
514/56 ;
514/266.24; 514/230.5; 514/309 |
International
Class: |
A61K 31/727 20060101
A61K031/727; A61K 31/517 20060101 A61K031/517; A61K 31/5355
20060101 A61K031/5355; A61K 31/4725 20060101 A61K031/4725; A61P
7/02 20060101 A61P007/02 |
Claims
1.-68. (canceled)
69. A method for preventing or treating thrombosis and thrombosis
related conditions in a mammal comprising the step of administering
to a mammal a therapeutically effective amount of a compound having
the formula (I): ##STR00150## wherein Y.sup.1 is NH; Y.sup.2 is CO,
each R.sup.1, R.sup.2 and R.sup.3 is independently selected from
the group consisting of H, C.sub.1-6alkyl, C.sub.2-6 alkenyl,
C.sub.2-6 alkynyl, --N(R.sup.7).sub.2, C.sub.1-6alkoxy, halogen,
C.sub.1-6haloalkyl, hydroxyC.sub.1-6alkyl, cyano, --C(O)R.sup.6 ,
C.sub.3-6cycloalkyl, C.sub.3-6cycloalkyl-C.sub.1-6alkyl, aryl and
arylC.sub.1-6alkyl, wherein each of said C.sub.1-6alkyl, C.sub.2-6
alkenyl, C.sub.2-6 alkynyl and aryl portions is optionally
substituted with from 1 to 3 substituents, each independently
selected from the group consisting of C.sub.1-6 alkyl, C.sub.2-6
alkenyl, C.sub.2-6 alkynyl, amino, C.sub.1-6alkoxy, halogen,
hydroxyl, cyano, oxo, thio, C.sub.3-6cycloalkyl, aryl and
heteroaryl; R.sup.4 is H or --(CH.sub.2).sub.mCO.sub.2H; R.sup.6 is
selected from the group consisting of hydroxy, C.sub.1-6alkyl,
C.sub.1-6alkoxy, heterocyclylC.sub.1-6alkoxy and
--N(R.sup.7).sub.2; each R.sup.7 is independently selected from the
group consisting of H, C.sub.1-6alkyl and arylC.sub.1-6alkyl or
optionally, two R.sup.7 groups attached to nitrogen are combined
with the nitrogen atom to form an azetidine, pyrrolidine,
piperidine or morpholine ring; wherein each of said C.sub.1-6 alkyl
and arylC.sub.1-6 alkyl is optionally substituted with from 1 to 3
substituents, each independently selected from the group consisting
of halogen, amino, hydroxyl, C.sub.1-6 alkoxy, cyano, C.sub.1-6
alkyl, C.sub.3-6 cycloalkyl, aryl, and heteroaryl; Ar.sup.1 is an
aromatic ring selected from the group consisting of benzene and
thiophene, each of which is optionally substituted with from 1-2
R.sup.8 substituents, Ar.sup.2 is an aromatic ring selected from
the group consisting of benzene, tetrazole and thiophene, each of
which is optionally substituted with from 1-2 R.sup.8 substituents,
each R.sup.8 is independently selected from the group consisting of
halogen, cyano, hydroxy, C.sub.1-6alkyl, C.sub.2-6alkenyl,
C.sub.2-6alkynyl, C.sub.1-6alkoxy, haloC.sub.1-6alkoxy,
C.sub.3-5cycloalkyl-C.sub.1-6alkoxy, --NR.sup.7,
--C(.dbd.NR.sup.8a)--N(R.sup.8b).sub.2, --C(O)R.sup.8a,
--O(CH.sub.2).sub.mOR.sup.8b, --(CH.sub.2).sub.mOR.sup.8b,
--O(CH.sub.2).sub.mN(R.sup.8b).sub.2 and
--(CH.sub.2).sub.mN(R.sup.8b).sub.2, each R.sup.8a is a member
independently selected from the group consisting of H, hydroxy,
C.sub.1-6alkyl, C.sub.1-6alkoxy and --NR.sup.7; each R.sup.8b is a
member independently selected from the group consisting of H,
C.sub.1-4 alkyl and C.sub.1-4alkanoyl, and optionally, two R.sup.8b
groups attached to nitrogen are combined with the nitrogen atom to
form an azetidine, pyrrolidine, piperidine or morpholine ring;
wherein each of said C.sub.1-4 alkyl and C.sub.1-4 alkanoyl is
optionally substituted with from 1 to 3 substituents, each
independently selected from the group consisting of halogen, amino,
hydroxyl, C.sub.1-6 alkoxy, cyano, C.sub.1-6 alkyl, C.sub.3-6
cycloalkyl, aryl, and heteroaryl; L.sup.1 is a linking group
selected from the group consisting of --CO-- and --CH.sub.2--,
L.sup.2 is a linking group selected from the group consisting of a
--C(R.sup.9).sub.2--, --C(R.sup.9).sub.2CH.sub.2-- and --CO--; each
R.sup.9 is independently selected from the group consisting of H,
C.sub.1-6alkyl, --(CH.sub.2).sub.mC(O)R.sup.6, --C(O)R.sup.6 and
heterocyclyl substituted with from 0 to 2 substituents selected
from the group halogen, cyano, C.sub.1-6alkyl, C.sub.2-6alkenyl,
C.sub.2-6alkynyl, C.sub.1-6alkoxy, haloC.sub.1-6alkyl, oxo and
thio; each subscript m is independently 1, 2 or 3, or
pharmaceutically acceptable salts thereof.
70. A method in accordance with claim 69, wherein said mammal is
human.
71. A method in accordance with claim 69, wherein said compound is
administered in combination with a second therapeutic agent.
72. A method in accordance with claim 71, wherein said second
therapeutic agent is useful for treating a condition or disorder
selected from the group consisting of acute myocardial infarction,
unstable angina, chronic stable angina, transient ischemic attacks,
strokes, peripheral vascular disease, preeclampsia/eclampsia, deep
venous thrombosis, embolism, disseminated intravascular coagulation
and thrombotic cytopenic purpura, thrombotic and restenotic
complications following invasive procedures resulting from
angioplasty, carotid endarterectorny, post CABG (coronary artery
bypass graft) surgery, vascular gram surgery, stent placements and
insertion of endovascular devices and prostheses.
73. A method in accordance with claim 71, wherein said second
therapeutic agent. selected from the group consisting of
antiplatelet compounds, anticoagulants, fibrinolytics,
anti-inflammatory compounds, cholesterol-lowering agents, blood
pressure-lowering agents and serotonin blockers.
74. A method in accordance with claim 71, wherein said second
therapeutic agent is an antiplatelet compound selected from the
group consisting of GPIIB-IIIa antagonists, aspirin,
phosphodiesterase III inhibitors and thromboxane A2 receptor
antagonists.
75. A method in accordance with claim 71, wherein said second
therapeutic agent is an anticoagulant selected from the group
consisting of thrombin inhibitors, Coumadin and heparin.
76. A method in accordance with claim 71, wherein said second
therapeutic agent is an anti-inflammatory compound selected from
the group consisting of non-steroidal anti-inflammatory agents,
cyclooxygenase-2 inhibitors and rheumatoid arthritis agents.
77. A method in accordance with claim 71, wherein said compound is
administered orally, parenterally or topically.
78.-83. (canceled)
84. A method in accordance with claim 71, wherein Ar.sup.1 is
benzene, which is optionally substituted with from 1-2 R.sup.8
substituents.
85. A method in accordance with claim 71, wherein Ar.sup.1 is
thiophene, which is optionally substituted with from 1-2 R.sup.8
substituents.
86. A method in accordance with claim 71, wherein Ar.sup.2 is
tetrazole, which is optionally substituted with from 1-2 R.sup.8
substituents.
87. A method in accordance with claim 71, wherein Ar.sup.2 is
benzene, which is optionally substituted with from 1-2 R.sup.8
substituents.
88. A method in accordance with claim 71, wherein Ar.sup.2 is
thiophene, which is optionally substituted with from 1-2 R.sup.8
substituents.
89. A method in accordance with claim 71, wherein at least one
R.sup.9 is --C(O)R.sup.6.
90. A method in accordance with claim 71, wherein at least one
R.sup.9 is --(CH.sub.2).sub.mC(O)R.sup.6.
91. A method in accordance with claim 71, wherein at least one
R.sup.9 is heterocyclyl.
92. A method in accordance with claim 71, wherein said compound has
the formula: ##STR00151## wherein the subscript n is an integer of
from 0 to 2.
93. A method in accordance with claim 71, wherein said compound has
the formula: ##STR00152## wherein the subscript n is an integer of
from 0 to 2.
94. A method in accordance with claim 71, wherein said compound has
the formula: ##STR00153## wherein the subscript n is an integer of
from 0 to 2.
95. A method in accordance with claim 71, wherein said compound is
selected from the group consisting of:
2-(5-chlorothiophen-2-yl)-2-(5-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-di-
hydroquinazolin-3(4H)-yl)thiophene-2-carboxamido)acetic acid;
2-(5-chlorothiophen-2-yl)-2-(5-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-di-
hydroquinazolin-3(4H)-yl)thiophene-3-carboxamido)acetic acid;
2-(4-(7-(4-fluorobenzylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)be-
nzamido)-3-(thiophen-2-yl)propanoic acid;
2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)b-
enzamido)-3-(thiophen-2-yl)propanoic acid;
N-((1H-tetrazol-5-yl)methyl)-4-(7-(4-fluorobenzylamino)-2,4-dioxo-1,2-dih-
ydroquinazolin-3(4H)-yl)benzamide;
N-((3-chlorophenyl)(1H-tetrazol-5-ypmethyl)-4-(6-fluoro-7-(methylamino)-2-
,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
N-((4-chlorophenyl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(methylamino)--
2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
2-(4-chlorophenyl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroqu-
inazolin-3(4H)-yl)benzamido)acetic acid;
2-(3-chlorophenyl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroqu-
inazolin-3(4H)-yl)benzamido)acetic acid;
N-((1H-tetrazol-5-yl)(m-tolyl)methyl)-4-(6-fluoro-7-(methylamino)-2,4-dio-
xo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)-N-(-
(3-methoxyphenyl)(1H-tetrazol-5-yl)methyl)benzamide;
4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)-N-(-
(3-fluorophenyl)(1H-tetrazol-5-yl)methyl)benzamide;
N-((2-chlorophenyl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(methylamino)--
2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
N-((3,4-dichlorophenyl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(methylami-
no)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
N-((3,5-dichlorophenyl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(methylami-
no)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
N-((1H-tetrazol-5-yl)(3-(trifluoromethoxy)phenyl)methyl)-4-(6-fluoro-7-(m-
ethylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
N-((1H-tetrazol-5-yl)(3-(trifluoromethyl)phenyl)methyl)-4-(6-fluoro-7-(me-
thylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
2-(5-chloro-N-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazoli-
n-3(4H)-yl)benzyl)thiophene-2-carboxamido)acetic acid;
3-(5-chlorothiophen-2-yl)-3-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-di-
hydroquinazolin-3(4H)-yl)benzylamino)propanoic acid;
N-((5-chlorothiophen-2-yl)methyl)-4-(7-(4-fluorobenzylamino)-2,4-dioxo-1,-
2-dihydroquinazolin-3(4H)-yl)benzamide;
2-(4-(7-(4-fluorobenzylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)be-
nzamido)-2-(thiophen-2-yl)acetic acid;
N-((5-chlorothiophen-2-yl)methyl)-4-(6-fluoro-7-(methylamino)-2,4-dioxo-1-
,2-dihydroquinazolin-3(4H)-yl)benzamide;
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-di-
hydroquinazolin-3(4H)-yl)benzamido)acetic acid;
2-(N-((5-chlorothiophen-2-yl)methyl)-4-(6-fluoro-7-(methylamino)-2,4-diox-
o-1,2-dihydroquinazolin-3(4H)-yl)benzamido)acetic acid;
N-((5-chlorothiophen-2-yl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(methyl-
amino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)b-
enzamido)-2-(thiophen-2-yl)acetic acid;
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-di-
hydroquinazolin-3(4H)-yl)benzamido)acetic acid;
2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3
(4H)-yl)benzamido)-2-(5-methylthiophen-2-yl)acetic acid;
2-(5-chlorothiophen-2-yl)-2-(4-(7-(methylamino)-2,4-dioxo-1,2-dihydroquin-
azolin-3(4H)-yl)benzamido)acetic acid;
N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)methyl)-
-4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benz-
amide;
N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-thiadiazol-3-yl)-
methyl)-4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)--
yl)benzamide; ethyl
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-di-
hydroquinazolin-3(4H)-yl)benzamido)acetate;
3-(5-chlorothiophen-2-yl)-3-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-di-
hydroquinazolin-3(4H)-yl)benzamido)propanoic acid;
N-((5-chlorothiophen-2-yl)(5-thioxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)meth-
yl)-4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)b-
enzamide;
2-(5-chlorothiophen-2-yl)-2-(4-(2,4-dioxo-1,2-dihydroquinazolin--
3(4H)-yl)benzamido)acetic acid;
N-((1H-tetrazol-5-yl)(3-(trifluoromethyl)phenyl)methyl)-4-(6-fluoro-7-(me-
thylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
2-morpholinoethyl
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-di-
hydroquinazolin-3(4H)-yl)benzamido)acetate;
N-(1-(5-chlorothiophen-2-yl)-2-hydroxyethyl)-4-(6-fluoro-7-(methylamino)--
2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
2-(5-chlorothiophen-2-yl)-2-(4-(6-iodo-2,4-dioxo-1,2-dihydroquinazolin-3(-
4H)-yl)benzamido)acetic acid;
2-(5-chlorothiophen-2-yl)-2-(4-(6-isopropoxy-2,4-dioxo-1,2-dihydroquinazo-
lin-3(4H)-yl)benzamido)acetic acid;
2-(5-chlorothiophen-2-yl)-2-(4-(2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)-
benzamido)acetic acid;
3-(N-((5-chlorothiophen-2-yl)methyl)-4-(6-fluoro-7-(methylamino)-2,4-diox-
o-1,2-dihydroquinazolin-3(4H)-yl)benzamido)propanoic acid;
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-di-
hydroquinazolin-3(4H)-yl)-2-methylbenzamido)acetic acid;
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-di-
hydroquinazolin-3(4H)-yl)-2-methoxybenzamido)acetic acid;
2-(2-chloro-4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3-
(4H)-yl)benzamido)-2-(5-chlorothiophen-2-yl)acetic acid;
2-(5-chlorothiophen-2-yl)-2-(2-fluoro-4-(6-fluoro-7-(methylamino)-2,4-dio-
xo-1,2-dihydroquinazolin-3(4H)-yl)benzamido)acetic acid;
N-(2-amino-1-(5-chlorothiophen-2-yl)-2-oxoethyl)-4-(6-fluoro-7-(methylami-
no)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
2-(5-chlorothiophen-2-yl)-2-(4-(7-fluoro-6-(methylamino)-2,4-dioxo-1,2-di-
hydroquinazolin-3(4H)-yl)benzamido)acetic acid;
N-((5-chlorothiophen-2-yl)(1H-tetrazol-5-yl)methyl)-4-(7-fluoro-6-(methyl-
amino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)methyl)-
-4-(7-fluoro-6-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benz-
amide;
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo--
1,2-dihydroquinazolin-3(4H)-yl)-3-methoxybenzamido)acetic acid;
N-((5-chlorothiophen-2-yl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(methyl-
amino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)-3-methoxybenzamide;
N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)methyl)-
-4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)-3-m-
ethoxybenzamide;
2-(1-((5-chlorothiophen-2-yl)methyl)-3-(4-(6-fluoro-7-(methylamino)-2,4-d-
ioxo-1,2-dihydroquinazolin-3(4H)-yl)phenyl)-1H-1,2,4-triazol-5-ylthio)acet-
ic acid;
2-(5-chlorothiophen-2-yl)-2-(3-(6-fluoro-7-(methylamino)-2,4-diox-
o-1,2-dihydroquinazolin-3(4H)-yl)benzamido)acetic acid;
N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)methyl)-
-4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benz-
amide,
(S)--N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-
-yl)methyl)-4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(-
4H)-yl)benzamide and
(R)--N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)me-
thyl)-4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl-
)benzamide.
Description
CROSS-REFERENCES TO RELATED APPLICATIONS
[0001] The present application is a Divisional of U.S. patent
application Ser. No. 11/856,616 filed Sep. 17, 2007 and claims
priority to U.S. 60/846,328 filed Sep. 20, 2006 herein incorporated
by reference in its entirety.
STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED
RESEARCH AND DEVELOPMENT
[0002] NOT APPLICABLE
REFERENCE TO A "SEQUENCE LISTING," A TABLE, OR A COMPUTER PROGRAM
LISTING APPENDIX SUBMITTED ON A COMPACT DISK.
[0003] NOT APPLICABLE
BACKGROUND OF THE INVENTION
[0004] Thrombotic complications are a major cause of death in the
industrialized world. Examples of these complications include acute
myocardial infarction, unstable angina, chronic stable angina,
transient ischemic attacks, strokes, peripheral vascular disease,
preeclampsia/eclampsia, deep venous thrombosis, embolism,
disseminated intravascular coagulation and thrombotic cytopenic
purpura. Thrombotic and restenotic complications also occur
following invasive procedures, e.g., angioplasty, carotid
endarterectomy, post CABG (coronary artery bypass graft) surgery,
vascular graft surgery, stent placements and insertion of
endovascular devices and protheses. It is generally thought that
platelet aggregates play a critical role in these events. Blood
platelets, which normally circulate freely in the vasculature,
become activated and aggregate to form a thrombus with disturbed
blood flow caused by ruptured atherosclerotic lesions or by
invasive treatments such as angioplasty, resulting in vascular
occlusion. Platelet activation can be initiated by a variety of
agents, e.g., exposed subendothelial matrix molecules such as
collagen, or by thrombin which is formed in the coagulation
cascade.
[0005] An important mediator of platelet activation and aggregation
is ADP (adenosine 5'-diphosphate) which is released from blood
platelets in the vasculature upon activation by various agents,
such as collagen and thrombin, and from damaged blood cells,
endothelium or tissues. Activation by ADP results in the
recruitment of more platelets and stabilization of existing
platelet aggregates. Platelet ADP receptors mediating aggregation
are activated by ADP and some of its derivatives and antagonized by
ATP (adenosine 5'-triphosphate) and some of its derivatives (Mills,
D. C. B. (1996) Thromb. Hemost. 76:835-856). Therefore, platelet
ADP receptors are members of the family of P2 receptors activated
by purine and/or pyrimidine nucleotides (King, B. F.,
Townsend-Nicholson, A. & Burnstock, G. (1998) Trends Pharmacol.
Sci. 19:506-514).
[0006] Recent pharmacological data using selective antagonists
suggests that ADP-dependent platelet aggregation requires
activation of at least two ADP receptors (Kunapuli, S. P. (1998),
Trends Pharmacol. Sci. 19:391-394; Kunapuli, S. P. & Daniel, J.
L. (1998) Biochem. J. 336:513-523; Jantzen, H. M. et al. (1999)
Thromb. Hemost. 81:111-117). One receptor appears to be identical
to the cloned P2Y.sub.1 receptor, mediates phospholipase C
activation and intracellular calcium mobilization and is required
for platelet shape change. The second platelet ADP receptor
important for aggregation mediates inhibition of adenylyl cyclase.
Molecular cloning of the gene or cDNA for this receptor
(P2Y.sub.12) has recently been reported (Hollopeter, G. et. al.
(2001) Nature 409:202-207). Based on its pharmacological and
signaling properties this receptor has been previously termed
P2Y.sub.ADP (Fredholm, B. B. et al. (1997) TIPS 18:79-82),
P2T.sub.AC (Kunapuli, S. P. (1998), Trends Pharmacol. Sci.
19:391-394) or P2Ycyc (Hechler, B. et al. (1998) Blood 92,
152-159).
[0007] Various directly or indirectly acting synthetic inhibitors
of ADP-dependent platelet aggregation with antithrombotic activity
have been reported. The orally active antithrombotic
thienopyridines ticlopidine and clopidogrel inhibit ADP-induced
platelet aggregation, binding of radiolabeled ADP receptor agonist
2-methylthioadenosine 5'-diphosphate to platelets, and other
ADP-dependent events indirectly, probably via formation of an
unstable and irreversible acting metabolite (Quinn, M. J. &
Fitzgerald, D. J. (1999) Circulation 100:1667-1667). Some purine
derivatives of the endogenous antagonist ATP, e.g., AR-C (formerly
FPL or ARL) 67085MX and AR-C69931MX, are selective platelet
[0008] ADP receptor antagonists which inhibit ADP-dependent
platelet aggregation and are effective in animal thrombosis models
(Humphries et al. (1995), Trends Pharmacol. Sci. 16, 179; Ingall,
A. H. et al. (1999) J. Med. Chem. 42, 213-230). Novel
triazolo[4,5-d]pyrimidine compounds have been disclosed as
P.sub.2T-antagonists (WO 99/05144). Tricyclic compounds as platelet
ADP receptor inhibitors have also been disclosed in WO 99/36425.
Piperazine derivatives are described in WO 02/098856. The target of
these antithrombotic compounds appears to be the platelet ADP
receptor mediating inhibition of adenylyl cyclase or
P2Y.sub.12.
[0009] Despite these compounds, there exists a need for more
effective platelet ADP receptor inhibitors. In particular, there is
a need for platelet ADP receptor inhibitors having antithrombotic
activity that are useful in the prevention and/or treatment of
cardiovascular diseases, particularly those related to
thrombosis.
BRIEF SUMMARY OF THE INVENTION
[0010] The present invention provides compounds having the
formula:
##STR00002##
and pharmaceutically acceptable salts thereof.
[0011] The symbol Y.sup.1 represents a member selected from the
group consisting of N, NH, O, CR.sup.5 and CH.sub.2.
[0012] The symbol Y.sup.2 is selected from the group consisting of
CO, CH.sub.2, CH and N.
[0013] Each symbol R.sup.1, R.sup.2 and R.sup.3 is independently
selected from the group consisting of H, C.sub.1-6alkyl, C.sub.2-6
alkenyl, C.sub.2-6 alkynyl, --NR.sup.7.sub.2, C.sub.1-6alkoxy,
halogen, C.sub.1-6haloalkyl, hydroxyC.sub.1-6alkyl, cyano,
--C(O)R.sup.6 , C.sub.3-6cycloalkyl,
C.sub.3-6cycloalkyl-C.sub.1-6alkyl, aryl and arylC.sub.1-6alkyl,
wherein each of said C.sub.1-6alkyl, C.sub.2-6 alkenyl, C.sub.2-6
alkynyl and aryl portions is optionally substituted with from 1 to
3 substituents, each independently selected from the group
consisting of C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6
alkynyl, amino, C.sub.1-6alkoxy, halogen, hydroxyl, cyano, oxo,
thio, C.sub.3-6cycloalkyl, aryl and heteroaryl.
[0014] The symbol R.sup.4 is H or --(CH.sub.2).sub.mCO.sub.2H.
[0015] The symbol R.sup.5 is selected from H, C.sub.1-6alkyl,
cyano, halogen, haloC.sub.1-6alkyl, aryl, arylC.sub.1-6alkyl and
--C(O)R.sup.6.
[0016] The symbol R.sup.6 is selected from the group consisting of
H, hydroxy, C.sub.1-6alkyl, C.sub.1-6alkoxy,
heterocyclylC.sub.1-6alkoxy and --NR.sup.7.sub.2.
[0017] Each symbol R.sup.7 is independently selected from the group
consisting of H, C.sub.1-6alkyl and arylC.sub.1-6alkyl or
optionally, two R.sup.7 groups attached to nitrogen are combined
with the nitrogen atom to form an azetidine, pyrrolidine,
piperidine or morpholine ring; wherein each of said C.sub.1-6 alkyl
and arylC.sub.1-6 alkyl is optionally substituted with from 1 to 3
substituents, each independently selected from the group consisting
of halogen, amino, hydroxyl, C.sub.1-6 alkoxy, cyano, C.sub.1-6
alkyl, C.sub.3-6 cycloalkyl, aryl, and heteroaryl.
[0018] Each symbol Ar.sup.1 and Ar.sup.2 represents an aromatic
ring selected from the group consisting of benzene, pyridine,
pyrazine, pyrimidine, tetrazole and thiophene, each of which is
optionally substituted with from 1-2 R.sup.8 substituents.
[0019] Each symbol R.sup.8 independently represents a member
selected from the group consisting of halogen, cyano, hydroxy,
C.sub.1-6alkyl, C.sub.2-6alkenyl, C.sub.2-6alkynyl,
C.sub.1-6alkoxy, haloC.sub.1-6alkyl, haloC.sub.1-6alkoxy,
C.sub.3-5cycloalkyl-C.sub.1-6alkoxy, --NR.sup.7,
--C(.dbd.NR.sup.8a)--N(R.sup.8b).sub.2, --C(O)R.sup.8a,
--O(CH.sub.2).sub.mOR.sup.8b, --(CH.sub.2).sub.mOR.sup.8b,
--O(CH.sub.2).sub.mN(R.sup.8b).sub.2 and
--(CH.sub.2).sub.mN(R.sup.8b).sub.2.
[0020] Each symbol R.sup.8a represents a member independently
selected from the group consisting of H, hydroxy, C.sub.1-6alkyl,
C.sub.1-6alkoxy and --NR.sup.7.
[0021] Each symbol R.sup.8b represents a member independently
selected from the group consisting of H, C.sub.1-4 alkyl and
C.sub.1-4alkanoyl, and optionally, two R.sup.8b groups attached to
nitrogen are combined with the nitrogen atom to form an azetidine,
pyrrolidine, piperidine or morpholine ring; wherein each of said
C.sub.1-4 alkyl and C.sub.1-4 alkanoyl is optionally substituted
with from 1 to 3 substituents, each independently selected from the
group consisting of halogen, amino, hydroxyl, C.sub.1-6 alkoxy,
cyano, C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl, aryl, and
heteroaryl.
[0022] The symbol L.sup.1 represents a linking group selected from
the group consisting of a bond, --CO--, --CH.sub.2--, --NHCO-- and
--CH.sub.2CO--.
[0023] The symbol L.sup.2 is a linking group selected from the
group consisting of a bond, --CR.sup.9.sub.2--,
--CR.sup.9.sub.2CH.sub.2-- and --CO--.
[0024] Each symbol R.sup.9 represents a member independently
selected from the group consisting of H, C.sub.1-6alkyl,
hydoxyalkyl, --(CH.sub.2).sub.mC(O)R.sup.6, --C(O)R.sup.6 and
heterocyclyl substituted with from 0 to 2 substituents selected
from the group halogen, cyano, C.sub.1-6alkyl, C.sub.2-6alkenyl,
C.sub.2-6alkynyl, C.sub.1-6alkoxy, haloC.sub.1-6alkyl, oxo and
thio.
[0025] Each subscript m is independently 1, 2 or 3.
[0026] In addition to the compounds provided herein, the present
invention further provides pharmaceutical compositions comprising
those compounds, as well as methods for the use of the compounds in
treating thrombosis as well as preventing the occurrence of a
secondary ischemic event.
BRIEF DESCRIPTION OF THE DRAWINGS
[0027] Not applicable.
DETAILED DESCRIPTION OF THE INVENTION
Definitions
[0028] The terms "treat", "treating" and "treatment" refer to any
method of alleviating or abrogating a disease or its attendant
symptoms.
[0029] The term "therapeutically effective amount" refers to that
amount of the compound being administered sufficient to prevent or
decrease the development of one or more of the symptoms of the
disease, condition or disorder being treated.
[0030] The term " inhibit" refers to the ability of a compound to
decrease the function, or activity, of the associated activity
(e.g., ADP). "Inhibition", as used herein in its various forms, is
meant to include antagonism and partial antagonism of the activity
associated with ADP. Inhibitors of ADP are compounds that, e.g.,
bind to, partially or totally block the enzyme's activity.
[0031] The term "compound" as used herein is intended to encompass
not only the specified molecular entity but also its
pharmaceutically acceptable, pharmacologically active derivatives,
including, but not limited to, salts, prodrug conjugates such as
esters and amides, metabolites, hydrates, solvates and the
like.
[0032] The term "composition" as used herein is intended to
encompass a product comprising the specified ingredients in the
specified amounts, as well as any product which results, directly
or indirectly, from combination of the specified ingredients in the
specified amounts. By "pharmaceutically acceptable" it is meant the
carrier, diluent or excipient must be compatible with the other
ingredients of the formulation and not deleterious to the recipient
thereof.
[0033] The "subject" is defined herein to include animals such as
mammals, including, but not limited to, primates (e.g., humans),
cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice and the
like. In some embodiments, the subject is a human.
[0034] As used herein, the term " ADP -mediated disease or
condition" and the like refers to a disease or condition
characterized by less than or greater than normal, ADP activity. A
ADP-mediated disease or condition is one in which modulation of ADP
results in some effect on the underlying condition or disease
(e.g., a ADP inhibitor or antagonist results in some improvement in
patient well-being in at least some patients).
[0035] The term "alkyl", by itself or as part of another
substituent, means, unless otherwise stated, a straight or branched
chain hydrocarbon radical, having the number of carbon atoms
designated (i.e. C.sub.1-.sub.8 means one to eight carbons).
Examples of alkyl groups include, but are not limited to methyl,
ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl,
n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. The term
"alkenyl" refers to an unsaturated alkyl group is one having one or
more double bonds directly attached to carbon radicals. Similarly,
the term "alkynyl" refers to an unsaturated alkyl group having one
or more triple bonds. Examples of such unsaturated alkyl groups
include, but are not limited to vinyl, 2-propenyl, crotyl,
2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl,
3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and the
higher homologs and isomers. The term "cycloalkyl" refers to
hydrocarbon rings having the indicated number of ring atoms (e.g.,
C.sub.3-6cycloalkyl) and being fully saturated or having no more
than one double bond between ring vertices. When "cycloalkyl" is
used in combination with "alkyl", as in C.sub.3-5 cycloalkyl-alkyl,
the cycloalkyl portion is meant to have from three to five carbon
atoms, while the alkyl portion is an alkylene moiety having from
one to three carbon atoms (e.g., --CH.sub.2--, --CH.sub.2CH.sub.2--
or --CH.sub.2CH.sub.2CH.sub.2--).
[0036] The terms "alkoxy," "alkylamino" and "alkylthio" (or
thioalkoxy) are used in their conventional sense, and refer to
those alkyl groups attached to the remainder of the molecule via an
oxygen atom, an amino group, or a sulfur atom, respectively. For
brevity, the term C.sub.1-6alkylamino is meant to include straight
chain, branched or cyclic alkyl groups or combinations thereof,
such as methyl, ethyl, 2-methylpropyl, cyclobutyl and
cyclopropylmethyl.
[0037] The terms "arylalkyl", "arylalkenyl" and "aryloxyalkyl"
refer to an aryl radical attached directly to an alkyl group, an
alkenyl group, or an oxygen which is attached to an alkyl group,
respectively. For brevity, aryl as part of a combined term as
above, is meant to include heteroaryl as well.
[0038] The terms "halo" or "halogen," by themselves or as part of
another substituent, mean, unless otherwise stated, a fluorine,
chlorine, bromine, or iodine atom. Additionally, terms such as
"haloalkyl," are meant to include monohaloalkyl and polyhaloalkyl.
For example, the term "C.sub.1-.sub.4 haloalkyl" is mean to include
trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl,
3-bromopropyl, and the like.
[0039] The terms "cycloalkyl" and "cycloalkenyl" refer to a
saturated hydrocarbon ring and includes bicyclic and polycyclic
rings. Similarly, cycloalkyl and cycloalkenyl groups having a
heteroatom (e.g. N, O, S or Si) in place of a carbon ring atom may
be referred to as "heterocycloalkyl", "heterocyclyl" and
"heterocycloalkylene," respectively. Accordingly, the term
"heterocyclyl" includes heteroaryl groups or rings. Examples of
cycloalkyl and heterocyclyl groups are, for example, cyclohexyl,
norbornyl, adamantyl, morpholinyl, thiomorpholinyl,
dioxothiomorpholinyl, pyridinyl, oxadiazolyl, thiadiazolyl,
tetrazoyl, thiazoyl and the like. The cycloalkyl and heterocyclyl
moieties may also be optionally substituted with halogen atoms, or
other groups such as nitro, alkyl, alkylamino, carboxyl, alkoxy,
aryloxy and the like. In some embodiments, cycloalkyl and
cycloalkenyl moieties are those having 3 to 12 carbon atoms in the
ring (e.g., cyclohexyl, cyclooctyl, norbornyl, adamantyl, and the
like). In some embodiments, heterocycloalkyl and
heterocycloalkylene moieties are those having 1 to 3 hetero atoms
in the ring (e.g., morpholinyl, thiomorpholinyl,
dioxothiomorpholinyl, piperidinyl and the like). Additionally, the
term "(cycloalkyl)alkyl" refers to a group having a cycloalkyl
moiety attached to an alkyl moiety. Examples are cyclohexylmethyl,
cyclohexylethyl and cyclopentylpropyl.
[0040] As used herein, the term "heteroatom" is meant to include
oxygen (O), nitrogen (N), sulfur (S) and silicon (Si).
[0041] The term "aryl" means, unless otherwise stated, a
polyunsaturated, typically aromatic, hydrocarbon group which can be
a single ring or multiple rings (up to three rings) which are fused
together or linked covalently. Exemplary aryl groups are phenyl (or
benzene), naphthyl, biphenyl and the like. The term "heteroaryl"
refers to aryl groups (or rings) that contain from one to five
heteroatoms selected from N, O, and S, wherein the nitrogen and
sulfur atoms are optionally oxidized, and the nitrogen atom(s) are
optionally quaternized. A heteroaryl group can be attached to the
remainder of the molecule through a heteroatom. Non-limiting
examples of heteroaryl groups include 1-pyrrolyl, 2-pyrrolyl,
3-pyrrolyl, 1-pyrazolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl,
pyrazinyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 3-isoxazolyl,
4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl,
2-furyl, 3-furyl, 2-thienyl (or 2-thiophenyl), 3-thienyl (or
3-thiophenyl), 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrazyl,
4-pyrazyl, 2-pyrimidyl, 4-pyrimidyl, 4-tetrazoyl, 5-tetrazoyl,
5-benzothiazolyl, purinyl, 2-benzimidazolyl, benzopyrazolyl,
5-indolyl, 1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl,
5-quinoxalinyl, 3-quinolyl, and 6-quinolyl. Substituents for each
of the above noted aryl and heteroaryl ring systems are selected
from the group of acceptable substituents described below.
[0042] The term "substituted" refers to the replacement of an atom
or a group of atoms of a compound with another atom or group of
atoms. For example, an atom or a group of atoms may be substituted
with one or more of the following substituents or groups: halo,
nitro, C.sub.1-C.sub.8alkyl, C.sub.1-C.sub.8alkylamino,
hydroxyC.sub.1-C.sub.8alkyl, halo C.sub.1-C.sub.8alkyl, carboxyl,
hydroxyl, C.sub.1-C.sub.8alkoxy, C.sub.1-C.sub.8alkoxy
C.sub.1-C.sub.8alkoxy, haloC.sub.1-C.sub.8alkoxy, thio
C.sub.1-C.sub.8alkyl, aryl, aryloxy, C.sub.3-C.sub.8cycloalkyl,
C.sub.3-C.sub.8cycloalkylC.sub.1-C.sub.8alkyl, aryl, heteroaryl,
arylC.sub.1-C.sub.8alkyl, heteroarylC.sub.1-C.sub.8alkyl,
C.sub.2-C.sub.8alkenyl containing 1 to 2 double bonds,
C.sub.2-C.sub.8alkynyl containing 1 to 2 triple bonds,
C.sub.2-C.sub.8alk(en)(yn)yl groups, cyano, formyl, oxo, thio,
C.sub.1-C.sub.8alkylcarbonyl, arylcarbonyl heteroarylcarbonyl,
carboxy, C.sub.1-C.sub.8alkoxycarbonyl, aryloxycarbonyl,
aminocarbonyl, C.sub.1-C.sub.8alkylaminocarbonyl,
C.sub.1-C.sub.8dialkylaminocarbonyl, arylaminocarbonyl,
diarylaminocarbonyl, arylC.sub.1-C.sub.8alkylaminocarbonyl,
aryloxy, haloC.sub.1-C.sub.8alkoxy, C.sub.2-C.sub.8alkenyloxy,
C.sub.2-C.sub.8alkynyloxy, arylC.sub.1-C.sub.8alkoxy,
aminoC.sub.1-C.sub.8alkyl,
C.sub.1-C.sub.8alkylaminoC.sub.1-C.sub.8alkyl,
C.sub.1-C.sub.8dialkylaminoC.sub.1-C.sub.8alkyl,
arylaminoC.sub.1-C.sub.8alkyl, amino, C.sub.1-C.sub.8dialkylamino,
arylamino, C.sub.1-C.sub.8alkylarylamino,
C.sub.1-C.sub.8alkylcarbonylamino, arylcarbonylamino, azido,
mercapto, C.sub.1-C.sub.8alkylthio, arylthio,
haloC.sub.1-C.sub.8alkylthio, thiocyano, isothiocyano,
C.sub.1-C.sub.8alkylsulfinyl, C.sub.1-C.sub.8alkylsulfonyl,
arylsulfinyl, arylsulfonyl, aminosulfonyl,
C.sub.1-C.sub.8alkylaminosulfonyl,
C.sub.1-C.sub.8dialkylaminosulfonyl and arylaminosulfonyl. When the
term "substituted" appears prior to a list of possible substituted
groups, it is intended that the term apply to every member of that
group.
[0043] The term "unsubstituted" refers to a native compound that
lacks replacement of an atom or a group of atoms.
[0044] The term "pharmaceutically acceptable salts" is meant to
include salts of the active compounds which are prepared with
relatively nontoxic acids or bases, depending on the particular
substituents found on the compounds described herein. When
compounds of the present invention contain relatively acidic
functionalities, base addition salts can be obtained by contacting
the neutral form of such compounds with a sufficient amount of the
desired base, either neat or in a suitable inert solvent. Examples
of pharmaceutically acceptable base addition salts include sodium,
potassium, calcium, ammonium, organic ammonium, zinc or magnesium
salt, or a similar salt. When compounds of the present invention
contain relatively basic functionalities, acid addition salts can
be obtained by contacting the neutral form of such compounds with a
sufficient amount of the desired acid, either neat or in a suitable
inert solvent. Examples of pharmaceutically acceptable acid
addition salts include those derived from inorganic acids like
hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic,
phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,
monohydrogensulfuric, hydriodic, or phosphorous acids and the like,
as well as the salts derived from relatively nontoxic organic acids
like acetic, propionic, isobutyric, malonic, benzoic, succinic,
suberic, fumaric, mandelic, phthalic, benzenesulfonic,
p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like.
Also included are salts of amino acids such as arginate and the
like, and salts of organic acids like glucuronic or galactunoric
acids and the like (see, for example, Berge, S. M., et al,
"Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977,
66, 1-19). Certain specific compounds of the present invention
contain both basic and acidic functionalities that allow the
compounds to be converted into either base or acid addition
salts.
[0045] The neutral forms of the compounds may be regenerated by
contacting the salt with a base or acid and isolating the parent
compound in the conventional manner. The parent form of the
compound differs from the various salt forms in certain physical
properties, such as solubility in polar solvents, but otherwise the
salts are equivalent to the parent form of the compound for the
purposes of the present invention.
[0046] In addition to salt forms, the present invention provides
compounds which are in a prodrug form. Prodrugs of the compounds
described herein are those compounds that readily undergo chemical
changes under physiological conditions to provide the compounds of
the present invention. Additionally, prodrugs can be converted to
the compounds of the present invention by chemical or biochemical
methods in an in vitro environment. For example, prodrugs can be
slowly converted to the compounds of the present invention when
placed in a transdermal patch reservoir with a suitable enzyme or
chemical reagent.
[0047] Certain compounds of the present invention can exist in
unsolvated forms as well as solvated forms, including hydrated
forms. In general, the solvated forms are equivalent to unsolvated
forms and are intended to be encompassed within the scope of the
present invention. Certain compounds of the present invention may
exist in multiple crystalline or amorphous forms. In general, all
physical forms are equivalent for the uses contemplated by the
present invention and are intended to be within the scope of the
present invention.
[0048] Certain compounds of the present invention possess
asymmetric carbon atoms (chiral centers) or double bonds; the
racemates, diastereomers, geometric isomers and individual isomers
(e.g., separate enantiomers) are all intended to be encompassed
within the scope of the present invention.
[0049] The compounds of the present invention may also contain
unnatural proportions of atomic isotopes at one or more of the
atoms that constitute such compounds. For example, the compounds
may be radiolabeled with radioactive isotopes, such as for example
tritium (.sup.3H), iodine-125 (.sup.125I), phosphorous-32
(.sup.32P) or carbon-14 (.sup.14C). All isotopic variations of the
compounds of the present invention, whether radioactive or not, are
intended to be encompassed within the scope of the present
invention.
General
Description of the Embodiments
[0050] Compounds
[0051] In view of the above, the present invention provides, in one
aspect, compounds having the formula:
provides compounds having the formula:
##STR00003##
and pharmaceutically acceptable salts thereof.
[0052] The symbol Y.sup.1 represents a member selected from the
group consisting of N, NH, O, CR.sup.5 and CH.sub.2.
[0053] The symbol Y.sup.2 is selected from the group consisting of
CO, CH.sub.2, CH and N.
[0054] Each symbol R.sup.1, R.sup.2 and R.sup.3 is independently
selected from the group consisting of H, C.sub.1-6alkyl, C.sub.2-6
alkenyl, C.sub.2-6 alkynyl, --NR.sup.7.sub.2, C.sub.1-6alkoxy,
halogen, C.sub.1-6haloalkyl, hydroxyC.sub.1-6alkyl, cyano,
--C(O)R.sup.6 , C.sub.3-6cycloalkyl,
C.sub.3-6cycloalkyl-C.sub.1-6alkyl, aryl and arylC.sub.1-6alkyl,
wherein each of said C.sub.1-6alkyl, C.sub.2-6 alkenyl, C.sub.2-6
alkynyl and aryl portions is optionally substituted with from 1 to
3 substituents, each independently selected from the group
consisting of C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6
alkynyl, amino, C.sub.1-6alkoxy, halogen, hydroxyl, cyano, oxo,
thio, C.sub.3-6cycloalkyl, aryl and heteroaryl.
[0055] The symbol R.sup.4 is H or --(CH.sub.2).sub.mCO.sub.2H.
[0056] The symbol R.sup.5 is selected from H, C.sub.1-6alkyl,
cyano, halogen, haloC.sub.1-6alkyl, aryl, arylC.sub.1-6alkyl and
--C(O)R.sup.6.
[0057] The symbol R.sup.6 is selected from the group consisting of
H, hydroxy, C.sub.1-6alkyl, C.sub.1-6alkoxy,
heterocyclylC.sub.1-6alkoxy and --NR.sup.7.sub.2.
[0058] Each symbol R.sup.7 is independently selected from the group
consisting of H, C.sub.1-6alkyl and arylC.sub.1-6alkyl or
optionally, two R.sup.7 groups attached to nitrogen are combined
with the nitrogen atom to form an azetidine, pyrrolidine,
piperidine or morpholine ring; wherein each of said C.sub.1-6 alkyl
and arylC.sub.1-6 alkyl is optionally substituted with from 1 to 3
substituents, each independently selected from the group consisting
of halogen, amino, hydroxyl, C.sub.1-6 alkoxy, cyano, C.sub.1-6
alkyl, C.sub.3-6 cycloalkyl, aryl, and heteroaryl.
[0059] Each symbol Ar.sup.1 and Ar.sup.2 represents an aromatic
ring selected from the group consisting of benzene, pyridine,
pyrazine, pyrimidine, tetrazole and thiophene, each of which is
optionally substituted with from 1-2 R.sup.8 substituents.
[0060] Each symbol R.sup.8 independently represents a member
selected from the group consisting of halogen, cyano, hydroxy,
C.sub.1-6alkyl, C.sub.2-6alkenyl, C.sub.2-6alkynyl,
C.sub.1-6alkoxy, haloC.sub.1-6alkyl, haloC.sub.1-6alkoxy,
C.sub.3-5cycloalkyl-C.sub.1-6alkoxy, --NR.sup.7,
--C(.dbd.NR.sup.8a)--N(R.sup.8b).sub.2, --C(O)R.sup.8a,
--O(CH.sub.2).sub.mOR.sup.8b, --(CH.sub.2).sub.mOR.sup.8b,
--O(CH.sub.2).sub.mN(R.sup.8b).sub.2 and
--(CH.sub.2).sub.mN(R.sup.8b).sub.2.
[0061] Each symbol R.sup.8a represents a member independently
selected from the group consisting of H, hydroxy, C.sub.1-6alkyl,
C.sub.1-6alkoxy and --NR.sup.7.
[0062] Each symbol R.sup.8b represents a member independently
selected from the group consisting of H, C.sub.1-4 alkyl and
C.sub.1-4alkanoyl, and optionally, two R.sup.8b groups attached to
nitrogen are combined with the nitrogen atom to form an azetidine,
pyrrolidine, piperidine or morpholine ring; wherein each of said
C.sub.1-4 alkyl and C.sub.1-4 alkanoyl is optionally substituted
with from 1 to 3 substituents, each independently selected from the
group consisting of halogen, amino, hydroxyl, C.sub.1-6 alkoxy,
cyano, C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl, aryl, and
heteroaryl.
[0063] The symbol L.sup.1 represents a linking group selected from
the group consisting of a bond, --CO--, --CH.sub.2--, --NHCO-- and
--CH.sub.2CO--.
[0064] The symbol L.sup.2 is a linking group selected from the
group consisting of a bond, --CR.sup.9.sub.2--,
--CR.sup.9.sub.2CH.sub.2-- and --CO--.
[0065] Each symbol R.sup.9 represents a member independently
selected from the group consisting of H, C.sub.1-6alkyl,
hydoxyalkyl, --(CH.sub.2).sub.mC(O)R.sup.6, --C(O)R.sup.6 and
heterocyclyl substituted with from 0 to 2 substituents selected
from the group halogen, cyano, C.sub.1-6alkyl, C.sub.2-6alkenyl,
C.sub.2-6alkynyl, C.sub.1-6alkoxy, haloC.sub.1-6alkyl, oxo and
thio.
[0066] Each subscript m is independently 1, 2 or 3.
[0067] In one group of embodiments, Y.sup.1 is NH and Y.sup.2 is
CO. In one group of embodiments, Y.sup.1 is O and Y.sup.2 is
CH.sub.2. In one group of embodiments, Y.sup.1 is CH and Y.sup.2 is
CH. In one group of embodiments, Y.sup.1 is N and Y.sup.2 is CH. In
one group of embodiments, Y.sup.1 is CH and Y.sup.2 is N. In one
group of embodiments, Y.sup.1 and Y.sup.2 are CH.sub.2
[0068] In another group of embodiments, each Ar.sup.1 and Ar.sup.2
are independently benzene, thiophene or tetrazole. In one group of
embodiments, Ar.sup.1 is benzene. In one group of embodiments,
Ar.sup.1 is thiophene. In one group of embodiments, Ar.sup.2 is
tetrazole. In one group of embodiments, Ar.sup.2 is benzene. In one
group of embodiments, Ar.sup.2 is thiophene. In any of the
embodiments herein, each of the groups comprising Ar.sup.1 and
Ar.sup.2 is optionally substituted with from 1-2 R.sup.8
substituents
[0069] In one group of embodiments, L.sup.1 is a bond. In one group
of embodiments, wherein L.sup.1 is --CO--. In one group of
embodiments, L.sup.1 is --CH.sub.2--. In one group of embodiments,
L.sup.1 is --NHCO--. In one group of embodiments, L.sup.1 is
--CH.sub.2CO--. In one group of embodiments, L.sup.2 is a bond. In
one group of embodiments, L.sup.2 is --CR.sup.9.sub.2--. In one
group of embodiments, L.sup.2 is --CR.sup.9.sub.2CH.sub.2--. In one
group of embodiments, L.sup.2 is --CO--.
[0070] In another group of embodiments, at least one R.sup.9 is H.
In one group of embodiments, at least one R.sup.9 is --C(O)R.sup.6.
In one group of embodiments, at least one R.sup.9 is
--(CH.sub.2).sub.mC(O)R.sup.6. In one group of embodiments, at
least one R.sup.9 is heterocyclyl. In one group of embodiments, at
least one R.sup.9 is independently selected from the group
consisting of:
##STR00004##
each of which is substituted with from 0 to 2 substituents selected
from the group halogen, cyano, C.sub.1-6alkyl, C.sub.2-6alkenyl,
C.sub.2-6 alkynyl, C.sub.1-6alkoxy, haloC.sub.1-6alkyl, oxo and
thio; and wherein the wavy line indicates the point of attachment
to the rest of the molecule. In one group of embodiments, at least
one R.sup.9 is independently selected from the group consisting
of:
##STR00005##
wherein the wavy line indicates the point of attachment to the rest
of the molecule.
[0071] In another group of embodiments, the compound has the
formula:
##STR00006##
wherein the subscript n is an integer of from 0 to 2.
[0072] In another group of embodiments, the compound has the
formula:
##STR00007##
wherein the subscript n is an integer of from 0 to 2.
[0073] In another group of embodiments, the compound has the
formula:
##STR00008##
wherein the subscript n is an integer of from 0 to 2.
[0074] In another group of embodiments, the compound has the
formula:
##STR00009##
wherein the subscript n is an integer of from 0 to 2.
[0075] In another group of embodiments, the compound has the
formula:
##STR00010##
wherein the subscript n is an integer of from 0 to 2.
[0076] In another group of embodiments, the compound has the
formula:
##STR00011##
wherein the subscript n is an integer of from 0 to 2.
[0077] In another group of embodiments, the compound has the
formula:
##STR00012##
wherein the subscript n is an integer of from 0 to 2.
[0078] In another group of embodiments, the compound has the
formula:
##STR00013##
wherein the subscript n is an integer of from 0 to 2.
[0079] In another group of embodiments, the compound has the
formula:
##STR00014##
wherein the subscript n is an integer of from 0 to 2.
[0080] In another group of embodiments, R.sup.1 is H, NHR.sup.7 or
halogen; R.sup.2 is H, C.sub.1-6alkyl, NHR.sup.7 or halogen;
R.sup.3 is H; R.sup.7 is C.sub.1-6alkyl or arylC.sub.1-6alkyl;
R.sup.8 is halogen or alkoxy; and n is 1. In one group of
embodiments, R.sup.1 is F; R.sup.2 is NHR.sup.7 and R.sup.7 is
CH.sub.3 or 4-fluorobenzyl.
[0081] Still other compounds of the present invention are:
[0082]
N-((5-chlorothiophen-2-yl)methyl)-4-(7-(4-fluorobenzylamino)-2,4-di-
oxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
[0083]
2-(4-(7-(4-fluorobenzylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-
-yl)benzamido)-2-(thiophen-2-yl)acetic acid;
[0084]
2-(4-(7-(4-fluorobenzylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-
-yl)benzamido)-3-(thiophen-2-yl)propanoic acid;
[0085]
N-((5-chlorothiophen-2-yl)methyl)-4-(6-fluoro-7-(methylamino)-2,4-d-
ioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
[0086]
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo--
1,2-dihydroquinazolin-3(4H)-yl)benzamido)acetic acid;
[0087]
2-(N-((5-chlorothiophen-2-yl)methyl)-4-(6-fluoro-7-(methylamino)-2,-
4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamido)acetic acid;
[0088]
N-((5-chlorothiophen-2-yl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(-
methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
[0089]
2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H-
)-yl)benzamido)-2-(thiophen-2-yl)acetic acid;
[0090]
2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H-
)-yl)benzamido)-3-(thiophen-2-yl)propanoic acid;
[0091]
N-((1H-tetrazol-5-yl)methyl)-4-(7-(4-fluorobenzylamino)-2,4-dioxo-1-
,2-dihydroquinazolin-3(4H)-yl)benzamide;
[0092]
N-((3-chlorophenyl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(methyla-
mino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
[0093]
N-((4-chlorophenyl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(methyla-
mino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
[0094]
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo--
1,2-dihydroquinazolin-3(4H)-yl)benzamido)acetic acid;
[0095]
2-(4-chlorophenyl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dih-
ydroquinazolin-3(4H)-yl)benzamido)acetic acid;
[0096]
2-(3-chlorophenyl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dih-
ydroquinazolin-3(4H)-yl)benzamido)acetic acid;
[0097]
4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)--
N-((5-methylthiophen-2-yl)(1H-tetrazol-5-yl)methyl)benzamide;
[0098] methyl
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e-
][1,3]oxazin-3(4H)-yl)benzamido)acetate;
[0099]
2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H-
)-yl)benzamido)-2-(5-methylthiophen-2-yl)acetic acid;
2-(5-chlorothiophen-2-yl)-2-(4-(7-(methylamino)-2,4-dioxo-1,2-dihydroquin-
azolin-3(4H)-yl)benzamido)acetic acid;
[0100]
N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)m-
ethyl)-4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-y-
l)benzamide;
[0101]
N-((1H-tetrazol-5-yl)(m-tolyl)methyl)-4-(6-fluoro-7-(methylamino)-2-
,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
[0102]
4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-y-
l)-N-((3-methoxyphenyl)(1H-tetrazol-5-yl)methyl)benzamide;
[0103]
4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-y-
l)-N-((3-fluorophenyl)(1H-tetrazol-5-yl)methyl)benzamide;
[0104]
N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-thiadiazol-3-yl)-
methyl)-4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)--
yl)benzamide;
[0105] ethyl
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-di-
hydroquinazolin-3(4H)-yl)benzamido)acetate;
[0106]
3-(5-chlorothiophen-2-yl)-3-(4-(6-fluoro-7-(methylamino)-2,4-dioxo--
1,2-dihydroquinazolin-3(4H)-yl)benzamido)propanoic acid;
[0107]
N-((2-chlorophenyl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(methyla-
mino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
[0108]
N-((3,4-dichlorophenyl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(met-
hylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
[0109]
N-((3,5-dichlorophenyl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(met-
hylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
[0110]
N-((1H-tetrazol-5-yl)(3-(trifluoromethoxy)phenyl)methyl)-4-(6-fluor-
o-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
[0111]
N-((5-chlorothiophen-2-yl)(5-thioxo-4,5-dihydro-1,2,4-oxadiazol-3-y-
l)methyl)-4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H-
)-yl)benzamide;
[0112]
2-(5-chlorothiophen-2-yl)-2-(4-(2,4-dioxo-1,2-dihydroquinazolin-3(4-
H)-yl)benzamido)acetic acid;
[0113]
N-((1H-tetrazol-5-yl)(3-(trifluoromethyl)phenyl)methyl)-4-(6-fluoro-
-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
[0114] 2-morpholinoethyl
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-di-
hydroquinazolin-3(4H)-yl)benzamido)acetate;
[0115]
N-(1-(5-chlorothiophen-2-yl)-2-hydroxyethyl)-4-(6-fluoro-7-(methyla-
mino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
[0116]
2-(5-chlorothiophen-2-yl)-2-(4-(6-iodo-2,4-dioxo-1,2-dihydroquinazo-
lin-3(4H)-yl)benzamido)acetic acid;
[0117]
2-(5-chlorothiophen-2-yl)-2-(4-(6-isopropoxy-2,4-dioxo-1,2-dihydroq-
uinazolin-3(4H)-yl)benzamido)acetic acid;
[0118]
2-(5-chlorothiophen-2-yl)-2-(4-(2,4-dioxo-1,2-dihydroquinazolin-3(4-
H)-yl)benzamido)acetic acid;
[0119]
3-(N-((5-chlorothiophen-2-yl)methyl)-4-(6-fluoro-7-(methylamino)-2,-
4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamido)propanoic
acid;
[0120]
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo--
1,2-dihydroquinazolin-3(4H)-yl)-2-methylbenzamido)acetic acid;
[0121]
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo--
1,2-dihydroquinazolin-3(4H)-yl)-2-methoxybenzamido)acetic acid;
[0122]
2-(2-chloro-4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinaz-
olin-3(4H)-yl)benzamido)-2-(5-chlorothiophen-2-yl)acetic acid;
[0123]
2-(5-chlorothiophen-2-yl)-2-(2-fluoro-4-(6-fluoro-7-(methylamino)-2-
,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamido)acetic acid;
[0124]
(S)--N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-
-yl)methyl)-4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(-
4H)-yl)benzamide;
[0125]
(R)--N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-
-yl)methyl)-4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(-
4H)-yl)benzamide;
[0126]
N-(2-amino-1-(5-chlorothiophen-2-yl)-2-oxoethyl)-4-(6-fluoro-7-(met-
hylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
[0127]
2-(5-chlorothiophen-2-yl)-2-(4-(7-fluoro-6-(methylamino)-2,4-dioxo--
1,2-dihydroquinazolin-3(4H)-yl)benzamido)acetic acid;
[0128]
N-((5-chlorothiophen-2-yl)(1H-tetrazol-5-yl)methyl)-4-(7-fluoro-6-(-
methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide;
[0129]
N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)m-
ethyl)-4-(7-fluoro-6-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-y-
l)benzamide;
[0130]
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo--
1,2-dihydroquinazolin-3(4H)-yl)-3-methoxybenzamido)acetic acid;
[0131]
N-((5-chlorothiophen-2-yl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(-
methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)-3-methoxybenzamide;
[0132]
N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)m-
ethyl)-4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-y-
l)-3-methoxybenzamide;
[0133]
N-((5-chlorothiophen-2-yl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(-
methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamide;
[0134]
2-(N-((5-chlorothiophen-2-yl)methyl)-4-(6-fluoro-7-(methylamino)-4--
oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamido)acetic acid;
[0135]
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-b-
enzo[e][1,3]oxazin-3(4H)-yl)benzamido)acetic acid;
[0136]
3-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-b-
enzo[e][1,3]oxazin-3(4H)-yl)benzamido)propanoic acid;
[0137]
2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-y-
l)benzamido)-2-phenylacetic acid;
[0138]
2-(5-chlorothiophen-2-yl)-2-(2-(4-(6-fluoro-7-(methylamino)-4-oxo-2-
H-benzo[e][1,3]oxazin-3(4H)-yl)phenyl)acetamido)-N,N-dimethylacetamide;
[0139]
N-(1-(5-chlorothiophen-2-yl)-2-oxo-2-(pyrrolidin-1-yl)ethyl)-2-(4-(-
6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)phenyl)acet-
amide;
[0140]
N-(2-amino-1-(5-chlorothiophen-2-yl)-2-oxoethyl)-2-(4-(6-fluoro-7-(-
methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)phenyl)acetamide;
[0141]
(R)-2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4-
H)-yl)benzamido)-3-phenylpropanoic acid;
[0142]
(S)-2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4-
H)-yl)benzamido)-3-phenylpropanoic acid;
[0143]
4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)--
N-((5-methylthiophen-2-yl)(1H-tetrazol-5-yl)methyl)benzamide;
[0144]
(S)-2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-4-oxo--
2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamido)acetic acid
[0145]
2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-y-
l)benzamido)-2-(thiophen-2-yl)acetic acid;
[0146]
2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-y-
l)benzamido)-3-(thiophen-2-yl)propanoic acid;
[0147]
(S)-2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4-
H)-yl)benzamido)-2-phenylacetic acid;
[0148] methyl
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e-
][1,3]oxazin-3(4H)-yl)benzamido)acetate;
[0149] ethyl
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e-
][1,3]oxazin-3(4H)-yl)benzamido)acetate;
[0150]
2-(5-chlorothiophen-2-yl)-2-(4-(8-methoxy-4-oxo-2H-benzo[e][1,3]oxa-
zin-3(4H)-yl)benzamido)acetic acid;
[0151]
2-(5-chlorothiophen-2-yl)-2-(4-(7-methyl-4-oxo-2H-benzo[e][1,3]oxaz-
in-3(4H)-yl)benzamido)acetic acid;
[0152]
N-(2-amino-1-(5-chlorothiophen-2-yl)-2-oxoethyl)-4-(6-fluoro-7-(met-
hylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamide;
[0153]
N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)m-
ethyl)-4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)b-
enzamide;
[0154]
2-(5-chlorothiophen-2-yl)-2-(4-(6-ethyl-4-oxo-2H-benzo[e][1,3]oxazi-
n-3(4H)-yl)benzamido)acetic acid;
[0155]
N-((5-chlorothiophen-2-yl)(1-methyl-1H-tetrazol-5-yl)methyl)-4-(6-f-
luoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamide;
[0156]
N-((5-chlorothiophen-2-yl)(2-methyl-2H-tetrazol-5-yl)methyl)-4-(6-f-
luoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamide;
[0157]
N-((5-chlorothiophen-2-yl)(5-thioxo-4,5-dihydro-1,2,4-oxadiazol-3-y-
l)methyl)-4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-y-
l)benzamide;
[0158] methyl
2-(5-chlorothiophen-2-yl)-2-(4-(6-methyl-4-oxo-2H-benzo[e][1,3]oxazin-3(4-
H)-yl)benzamido)acetate;
[0159]
2-(5-chlorothiophen-2-yl)-2-(4-(6-methyl-4-oxo-2H-benzo[e][1,3]oxaz-
in-3(4H)-yl)benzamido)acetic acid;
[0160]
2-(5-chlorothiophen-2-yl)-2-(4-(7-ethyl-4-oxo-2H-benzo[e][1,3]oxazi-
n-3(4H)-yl)benzamido)acetic acid;
[0161]
3-(5-chlorothiophen-2-yl)-3-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-b-
enzo[e][1,3]oxazin-3(4H)-yl)benzamido)propanoic acid;
[0162]
N-((3-chlorophenyl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(methyla-
mino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamide;
[0163]
2-(3-chlorophenyl)-2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e]-
[1,3]oxazin-3(4H)-yl)benzamido)acetic acid;
[0164]
N-((5-chlorothiophen-2-yl)(thiazol-2-yl)methyl)-4-(6-fluoro-7-(meth-
ylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamide;
[0165]
2-(5-chlorothiophen-2-yl)-2-(4-(4-oxo-2H-benzo[e][1,3]oxazin-3(4H)--
yl)benzamido)acetic acid;
[0166]
2-(5-chlorothiophen-2-yl)-2-(4-(7-(4-fluorobenzylamino)-4-oxo-2H-be-
nzo[e][1,3]oxazin-3(4H)-yl)benzamido)acetic acid;
[0167]
N-((5-chlorothiophen-2-yl)(4-methylthiazol-2-yl)methyl)-4-(6-fluoro-
-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamide;
[0168]
3-(N-((5-chlorothiophen-2-yl)methyl)-4-(6-fluoro-7-(methylamino)-4--
oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamido)propanoic acid;
[0169]
N-((5-chlorothiophen-2-yl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(-
methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamide;
[0170]
(S)--N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-
-yl)methyl)-4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-
-yl)benzamide;
[0171]
(R)--N-((5-chlorothiophen-2-yl)(1H-tetrazol-5-yl)methyl)-4-(6-fluor-
o-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamide;
[0172]
2-(5-chlorothiophen-2-yl)-2-(4-(7-fluoro-6-(methylamino)-1-oxoisoqu-
inolin-2(1H)-yl)benzamido)acetic acid;
[0173]
N-((5-chlorothiophen-2-yl)(1H-tetrazol-5-yl)methyl)-4-(7-fluoro-6-(-
methylamino)-1-oxoisoquinolin-2(1H)-yl)benzamide;
[0174]
2-(5-chlorothiophen-2-yl)-2-(4-(7-fluoro-6-methoxy-1-oxoisoquinolin-
-2(1H)-yl)benzamido)acetic acid;
[0175]
2-(5-chlorothiophen-2-yl)-2-(4-(6-ethyl-1-oxoisoquinolin-2(1H)-yl)b-
enzamido)acetic acid;
[0176]
2-(4-(6-carbamoyl-1-oxoisoquinolin-2(1H)-yl)benzamido)-2-(5-chlorot-
hiophen-2-yl)acetic acid;
[0177]
2-(5-chlorothiophen-2-yl)-2-(4-(6-cyano-1-oxoisoquinolin-2(1H)-yl)b-
enzamido)acetic acid;
[0178]
2-(5-chlorothiophen-2-yl)-2-(4-(1-oxoisoquinolin-2(1H)-yl)benzamido-
)acetic acid;
[0179]
2-(4-(1-oxoisoquinolin-2(1H)-yl)benzamido)-2-(thiophen-2-yl)acetic
acid;
[0180]
2-(5-chlorothiophen-2-yl)-2-(4-(6-cyano-1-oxoisoquinolin-2(1H)-yl)b-
enzamido)acetic acid;
[0181]
4-(4-carbamoyl-1-oxoisoquinolin-2(1H)-yl)benzamido)-2-(5-chlorothio-
phen-2-yl)acetic acid;
[0182]
2-(5-chlorothiophen-2-yl)-2-(4-(4-cyano-1-oxoisoquinolin-2(1H)-yl)b-
enzamido)acetic acid;
[0183]
2-(5-chlorothiophen-2-yl)-2-(4-(6-methoxy-1-oxoisoquinolin-2(1H)-yl-
)benzamido)acetic acid;
[0184]
2-(4-(4-bromo-1-oxoisoquinolin-2(1H)-yl)benzamido)-2-(5-chlorothiop-
hen-2-yl)acetic acid;
[0185]
2-(5-chloro-N-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroqui-
nazolin-3(4H)-yl)benzyl)thiophene-2-carboxamido)acetic acid;
[0186]
3-(5-chlorothiophen-2-yl)-3-(4-(6-fluoro-7-(methylamino)-2,4-dioxo--
1,2-dihydroquinazolin-3(4H)-yl)benzylamino)propanoic acid;
[0187]
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-b-
enzo[e][1,3]oxazin-3(4H)-yl)benzylamino)acetic acid;
[0188]
3-(5-chlorothiophen-2-yl)-3-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-b-
enzo[e][1,3]oxazin-3(4H)-yl)benzylamino)propanoic acid;
[0189]
1-((5-chlorothiophen-2-yl)methyl)-3-(4-(7-(4-fluorobenzylamino)-2,4-
-dioxo-1,2-dihydroquinazolin-3(4H)-yl)phenyl)urea;
[0190]
2-(3-(4-(7-(4-fluorobenzylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(-
4H)-yl)phenyl)ureido)-2-(thiophen-2-yl)acetic acid;
[0191]
2-(3-(4-(7-(4-fluorobenzylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(-
4H)-yl)phenyl)ureido)-3-(thiophen-2-yl)propanoic acid;
[0192]
2-(5-chlorothiophen-2-yl)-2-(3-(4-(6-fluoro-7-(methylamino)-2,4-dio-
xo-1,2-dihydroquinazolin-3(4H)-yl)phenyl)ureido)acetic acid;
[0193]
2-(5-chlorothiophen-2-yl)-2-(3-(6-fluoro-7-(methylamino)-2,4-dioxo--
1,2-dihydroquinazolin-3(4H)-yl)benzamido)acetic acid;
[0194]
2-(5-chlorothiophen-2-yl)-2-(5-(6-fluoro-7-(methylamino)-2,4-dioxo--
1,2-dihydroquinazolin-3(4H)-yl)thiophene-2-carboxamido)acetic
acid;
[0195]
2-(5-chlorothiophen-2-yl)-2-(5-(6-fluoro-7-(methylamino)-2,4-dioxo--
1,2-dihydroquinazolin-3(4H)-yl)thiophene-3-carboxamido)acetic
acid;
[0196]
2-(5-chlorothiophen-2-yl)-2-(2-(4-(6-fluoro-7-(methylamino)-4-oxo-2-
H-benzo[e][1,3]oxazin-3(4H)-yl)phenyl)acetamido)acetic acid;
[0197]
2-(5-chlorothiophen-2-yl)-2-(4-(7-fluoro-6-(methylamino)-1-oxoisoqu-
inolin-2(1H)-yl)benzylamino)acetic acid;
[0198]
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-4-oxoquina-
zolin-3(4H)-yl)benzamido)acetic acid;
[0199]
2-(5-chlorothiophen-2-yl)-2-(4-(7-fluoro-6-(methylamino)-1-oxophtha-
lazin-2(1H)-yl)benzamido)acetic acid;
[0200] methyl
2-(5-chlorothiophen-2-yl)-2-(2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benz-
o[e][1,3]oxazin-3(4H)-yl)phenyl)acetamido)acetate;
[0201]
2-(5-chlorothiophen-2-yl)-2-(4-(6-ethyl-1-oxo-3,4-dihydroisoquinoli-
n-2(1H)-yl)benzamido)acetic acid; and
[0202]
2-(1-((5-chlorothiophen-2-yl)methyl)-3-(4-(6-fluoro-7-(methylamino)-
-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)phenyl)-1H-1,2,4-triazol-5-ylthi-
o)acetic acid.
[0203] Consistent with the practice of those of skill in the art,
unlabeled bonds (e.g., those with an unlabeled terminus) are meant
to illustrate methyl (CH.sub.3) groups.
[0204] General Synthetic Schemes
##STR00015##
[0205] The starting materials and reagents used in preparing these
compounds generally are either available from commercial suppliers,
such as Aldrich Chemical Co., or are prepared by methods known to
those skilled in the art following procedures set forth in
references such as Fieser and Fieser's Reagents for Organic
Synthesis; Wiley & Sons: New York, 1967-2004, Volumes 1-22;
Rodd's Chemistry of Carbon Compounds, Elsevier Science Publishers,
1989, Volumes 1-5 and Supplementals; and Organic Reactions, Wiley
& Sons: New York, 2005, Volumes 1-65. The following synthetic
reaction schemes are merely illustrative of some methods by which
the compounds of the present invention can be synthesized, and
various modifications to these synthetic reaction schemes can be
made and will be suggested to one skilled in the art having
referred to the disclosure contained in this Application.
[0206] The starting materials and the intermediates of the
synthetic reaction schemes can be isolated and purified if desired
using conventional techniques, including but not limited to,
filtration, distillation, crystallization, chromatography, and the
like. Such materials can be characterized using conventional means,
including physical constants and spectral data.
[0207] Unless specified to the contrary, the reactions described
herein preferably are conducted under an inert atmosphere at
atmospheric pressure at a reaction temperature range of from about
-78.degree. C. to about 150.degree. C., more preferably from about
0.degree. C. to about 125.degree. C., and most preferably and
conveniently at about room (or ambient) temperature, e.g., about
20.degree. C.
[0208] Scheme A describes a method of preparing a compound of
formula I wherein Y.sup.1 is O and the remaining symbols are as
defined hereinbefore.
##STR00016##
[0209] A compound of Formula I can be prepared by reacting a
salicylic acid with acetic anhydride followed by acid chloride
formation with oxalyl chloride which was converted to amides by
reaction with various benzoate compounds. Transesterification under
basic conditions to affords salicylamides 3. The intermediate
salicylamides 3 were cyclized with paraformaldehyde to bicyclic
compounds 4. The ester of compound 4 can be hydrolyzed by
procedures known to one skilled in the art to yield free carboxylic
acid. For example, a method of hydrolysis can be carried out using
aqueous lithium hydroxide with a cosolvent, typically
tetrahydrofuran or dioxane. Formation of the amide linkate can be
accomplished by treating the carboxylic acid 5 a coupling reagent,
for example, HATU in the presence of a tertiary amine base,
followed by addition of the amine to be coupled after a suitable
period of time to give amide 6.
[0210] Scheme B illustrates another preparation of compounds of
formula I with varying Ar.sup.1, Ar.sup.2, L.sup.1 and L.sup.2
groups.
##STR00017##
[0211] A compound of Formula I with varying Ar.sup.1, Ar.sup.2,
L.sup.1 and L.sup.2 groups can be prepared by first synthesizing
the common intermediate 5 in 4 steps (Scheme A). The acid chloride
from Scheme A can be treated with p-methoxybenzylamine followed by
cyclization with paraformaldehyde to afford the bicyclic ring
system. The p-methoxybenzyl functionality can then be cleaved under
acidic conditions to afford common intermediate 5. A variety of
halo-substituted compounds can be coupled with 5 using a copper
catalyzed coupling to give 6 followed by deprotection of any
functional groups if necessary. Completion of the synthesis can be
accomplished by coupling to form an amide using the conditions in
scheme A, or by reductive amination to give a reduced linkage. The
examples provided in detail below illustrate compounds prepared by
the general methods provided.
##STR00018##
[0212] A compound of Formula I with varying Ar.sup.1, Ar.sup.2,
L.sup.1 and L.sup.2 groups can be prepared by starting with a
suitable anthranilic ester 1 and a suitably substituted Ar.sup.1
aniline, 2, and treating one of the compounds with excess phosgene
and a tertiary amine base if needed. After removal of excess
phosgene the activated intermediate is taken up in a solvent such
as dichloromethane, and the remaining reagent and a tertiary amine
base is added. The cyclized product can then be further elaborated
using conditions described in previous schemes to the final
compound 4.
[0213] Compositions
[0214] In another aspect of the invention, pharmaceutical
compositions are provided in which compounds of formulae I, II,
III, IV, V, VI or VII, alone or in combination, are combined with a
pharmaceutically acceptable carrier.
[0215] The pharmaceutical compositions of the invention may be in
the form of solutions or suspensions. In the management of
thrombotic disorders the compounds or pharmaceutical compositions
of the invention may also be in such forms as, for example,
tablets, capsules or elixirs for oral administration,
suppositories, sterile solutions or suspensions or injectable
administration, and the like, or incorporated into shaped
articles.
[0216] Typical adjuvants which may be incorporated into tablets,
capsules and the like include, but are not limited to, binders such
as acacia, corn starch or gelatin, and excipients such as
microcrystalline cellulose, disintegrating agents like corn starch
or alginic acid, lubricants such as magnesium stearate, sweetening
agents such as sucrose or lactose, or flavoring agents. When a
dosage form is a capsule, in addition to the above materials it may
also contain liquid carriers such as water, saline, or a fatty oil.
Other materials of various types may be used as coatings or as
modifiers of the physical form of the dosage unit. Sterile
compositions for injection can be formulated according to
conventional pharmaceutical practice. For example, dissolution or
suspension of the active compound in a vehicle such as an oil or a
synthetic fatty vehicle like ethyl oleate, or into a liposome may
be desired. Buffers, preservatives, antioxidants and the like can
be incorporated according to accepted pharmaceutical practice.
[0217] Additionally, dosage formulations of compounds of formulae
I, II, III, IV, V, VI or VII, or pharmaceutical compositions
containing a compound of the invention, to be used for therapeutic
administration must be sterile. Sterility can be readily
accomplished by filtration through sterile membranes such as 0.2
micron membranes, or by other conventional methods. Formulations
typically will be stored in a solid form, preferably in a
lyophilized form. While the preferred route of administration is
orally, the dosage formulations of compounds of formulae I, II,
III, IV, V, VI or VII, or pharmaceutical compositions of the
invention may also be administered by injection, intravenously
(bolus and/or infusion), subcutaneously, intramuscularly,
colonically, rectally, nasally, transdermally or intraperitoneally.
A variety of dosage forms may be employed as well including, but
not limited to, suppositories, implanted pellets or small
cylinders, aerosols, oral dosage formulations and topical
formulations such as ointments, drops and dermal patches. The
compounds of formulae I, II, III, IV, V, VI or VII, and
pharmaceutical compositions of the invention may also be
incorporated into shapes and articles such as implants which may
employ inert materials such biodegradable polymers or synthetic
silicones as, for example, SILASTIC, silicone rubber or other
polymers commercially available. The compounds and pharmaceutical
compositions of the invention may also be provided in the form of
liposome delivery systems, such as small unilamellar vesicles,
large unilamellar vesicles and multilamellar vesicles. Liposomes
can be formed from a variety of lipids, such as cholesterol,
stearylamine or phosphatidylcholines, used methods well known to
one of skill in the art.
Methods of Treatment/Administration
[0218] In yet another aspect, the present invention provides
methods for preventing or treating thrombosis in a mammal by
administering to the mammal a therapeutically effective amount of a
compound of formulae I, II, III, IV, V, VI or VII, alone or as part
of a pharmaceutical composition of the invention as described
above. Compounds of formulae I, II, III, IV, V, VI or VII, and
pharmaceutical compositions of the invention containing a compound
of formulae I, II, III, IV, V, VI or VII, of the invention are
suitable for use alone or as part of a multi-component treatment
regimen for the prevention or treatment of cardiovascular diseases,
particularly those related to thrombosis. For example, a compound
or pharmaceutical composition of the invention may be used as a
drug or therapeutic agent for any thrombosis, particularly a
platelet-dependent thrombotic indication, including, but not
limited to, acute myocardial infarction, unstable angina, chronic
stable angina, transient ischemic attacks, strokes, peripheral
vascular disease, preeclampsia/eclampsia, deep venous thrombosis,
embolism, disseminated intravascular coagulation and thrombotic
cytopenic purpura, thrombotic and restenotic complications
following invasive procedures, e.g., angioplasty, carotid
endarterectomy, post CABG (coronary artery bypass graft) surgery,
vascular graft surgery, stent placements and insertion of
endovascular devices and protheses.
[0219] Compounds and pharmaceutical compositions of the invention
may also be used as part of a multi-component treatment regimen in
combination with other therapeutic or diagnostic agents in the
prevention or treatment of thrombosis in a mammal. In certain
preferred embodiments, compounds or pharmaceutical compositions of
the invention may be coadministered along with other compounds
typically prescribed for these conditions according to generally
accepted medical practice such as anticoagulant agents,
thrombolytic agents, or other antithrombotics, including platelet
aggregation inhibitors, tissue plasminogen activators, urokinase,
prourokinase, streptokinase, heparin, aspirin, or warfarin. Still
other agents that can be administered with the compounds of the
present invention include antiplatelet compounds, fibrinolytics,
anti-inflammatory compounds, cholesterol-lowering agents,
blood-pressure-lowering agents and serotonin blockers. Suitable
antiplatelet compounds include GPIIB-IIIa antagonists, aspirin,
phosphodiesterase III inhibitors and thromboxane A2 receptor
antagonists. Suitable anticoagulants include thrombin inhibitors,
fXa inhibitors, coumadin (Warfarin), heparin and Lovenox.RTM..
Suitable anti-inflammatory compounds include non-steroidal
anti-inflammatory agents, cyclooxygenase-2 inhibitors and
rheumatoid arthritis agents. Coadministrations of these agents with
the compounds of the invention may also allow for application of
reduced doses of the thrombolytic agents and therefore minimize
potential hemorrhagic side-effects. Compounds and pharmaceutical
compositions of the invention may also act in a synergistic fashion
to prevent reocclusion following a successful thrombolytic therapy
and/or reduce the time to reperfusion.
[0220] In related methods, the compounds of the invention are
useful for the prevention of a secondary ischemic event. In these
methods, compounds of the invention or their pharmaceutical
compositions are administered to a patient who has suffered a
primary ischemic event in an amount sufficient to prevent or reduce
the likely occurrence of a secondary event. Generally, the primary
and/or secondary ischemic event is selected from myocardial
infraction, stable or unstable angina, acute reocclusion after
percutaneous transluminal coronary angioplasty, restenosis,
thrombotic stroke, transient ischemic attack, reversible ischemic
neurological deficit and intermittent claudication.
[0221] The compounds and pharmaceutical compositions of the
invention may be utilized in vivo, ordinarily in mammals such as
primates, (e.g., humans), sheep, horses, cattle, pigs, dogs, cats,
rats and mice, or in vitro. The biological properties, as defined
above, of a compound or a pharmaceutical composition of the
invention can be readily characterized by methods that are well
known in the art such as, for example, by in vivo studies to
evaluate antithrombotic efficacy, and effects on hemostasis and
hematological parameters.
[0222] Subjects (typically mammalian) in need of treatment using
the compounds or pharmaceutical compositions of the invention may
be administered dosages that will provide optimal efficacy. The
dose and method of administration will vary from subject to subject
and be dependent upon such factors as the type of mammal being
treated, its sex, weight, diet, concurrent medication, overall
clinical condition, the particular compound of formulae I, II, III,
IV, V, VI or VII, employed, the specific use for which the compound
or pharmaceutical composition is employed, and other factors which
those skilled in the medical arts will recognize.
[0223] Therapeutically effective dosages may be determined by
either in vitro or in vivo methods. For each particular compound or
pharmaceutical composition of the invention, individual
determinations may be made to determine the optimal dosage
required. The range of therapeutically effective dosages will be
influenced by the route of administration, the therapeutic
objectives and the condition of the patient. For injection by
hypodermic needle, it may be assumed the dosage is delivered into
the bodily fluids. For other routes of administration, the
absorption efficiency must be individually determined for each
compound by methods well known in pharmacology. Accordingly, it may
be necessary for the therapist to titer the dosage and modify the
route of administration as required to obtain the optimal
therapeutic effect.
[0224] The determination of effective dosage levels, that is, the
dosage levels necessary to achieve the desired result, i.e.,
platelet ADP receptor inhibition, will be readily determined by one
skilled in the art. Typically, applications of a compound or
pharmaceutical composition of the invention are commenced at lower
dosage levels, with dosage levels being increased until the desired
effect is achieved. The compounds and compositions of the invention
may be administered orally in an effective amount within the dosage
range of about 0.01 to 1000 mg/kg in a regimen of single or several
divided daily doses. If a pharmaceutically acceptable carrier is
used in a pharmaceutical composition of the invention, typically,
about 5 to 500 mg of a compound of formulae I, II, III, IV, V, VI
or VII, is combined with a pharmaceutically acceptable carrier as
called for by accepted pharmaceutical practice including, but not
limited to, a physiologically acceptable vehicle, carrier,
excipient, binder, preservative, stabilizer, dye, flavor, etc. The
amount of active ingredient in these compositions is such that a
suitable dosage in the range indicated is obtained.
[0225] The following preparations and examples are given to enable
those skilled in the art to more clearly understand and to practice
the present invention. They should not be considered as limiting
the scope of the invention, but merely as being illustrative and
representative thereof.
EXAMPLES
[0226] The starting materials and the intermediates of the
synthetic reaction schemes can be isolated and purified if desired
using conventional techniques, including but not limited to,
filtration, distillation, crystallization, chromatography, and the
like. Such materials can be characterized using conventional means,
including physical constants and spectral data.
[0227] Unless specified to the contrary, the reactions described
herein preferably are conducted under an inert atmosphere at
atmospheric pressure at a reaction temperature range of from about
-78.degree. C. to about 150.degree. C., more preferably from about
0.degree. C. to about 125.degree. C., and most preferably and
conveniently at about room (or ambient) temperature, e.g., about
20.degree. C.
[0228] Referring to the examples that follow, compounds of the
present invention were synthesized using the methods described
herein, or other methods, which are well known in the art.
[0229] The compounds and/or intermediates were characterized by
high performance liquid chromatography (HPLC) using a Waters
Alliance chromatography system with a 2695 Separation Module
(Milford, Mass.). The analytical columns were C-18 SpeedROD RP-18E
Columns from Merck KGaA (Darmstadt, Germany). The enantiomeric
purity of the material was measured by chiral HPLC using an
(R,R)-ULMO column (25 cm, .times.4.6 mm, 5 um) from Regis
Technologies eluting with 75/25 hexane/ethanol with 25 mM ammonium
acetate and 1% triethyl amine. Alternately, characterization was
performed using a Waters Unity (UPLC) system with Waters Acquity
UPLC BEH C-18 2.1 mm.times.15 mm columns. A gradient elution was
used, typically starting with 5% acetonitrile/95% water and
progressing to 95% acetonitrile over a period of 5 minutes for the
Alliance system and 1 minute for the Acquity system. All solvents
contained 0.1% trifluoroacetic acid (TFA). Compounds were detected
by ultraviolet light (UV) absorption at either 220 or 254 nm. HPLC
solvents were from EMD Chemicals, Inc. (Gibbstown, N.J.). In some
instances, purity was assessed by thin layer chromatography (TLC)
using glass backed silica gel plates, such as, for example, EMD
Silica Gel 60 2.5 cm.times.7.5 cm plates. TLC results were readily
detected visually under ultraviolet light, or by employing well
known iodine vapor and other various staining techniques.
[0230] Mass spectrometric analysis was performed on one of two
Agilent 1100 series LCMS instruments with acetonitrile/water as the
mobile phase. One system using TFA as the modifier and measures in
positive ion mode and the other uses either formic acid or ammonium
acetate and measures in both positive and negative ion modes.
[0231] Nuclear magnetic resonance (NMR) analysis was performed on
some of the compounds with a Varian 400 MHz NMR (Palo Alto,
Calif.). The spectral reference was either TMS or the known
chemical shift of the solvent. Some compound samples were run at
elevated temperatures (e.g., 75.degree. C.) to promote increased
sample solubility.
[0232] Preparative separations were carried out using either an
Sq16x or an Sg100c chromatography system and prepackaged silica gel
columns all purchased from Teledyne Isco, (Lincoln, Nebr.).
Alternately, compounds and intermediates were purified by flash
column chromatography using silica gel (230-400 mesh) packing
material, or by HPLC using a C-18 reversed phase column. Typical
solvents employed for the Isco systems and flash column
chromatography were dichloromethane, methanol, ethyl acetate,
hexane, acetone, aqueous hydroxyamine and triethyl amine. Typical
solvents employed for the reverse phase HPLC were varying
concentrations of acetonitrile and water with 0.1% trifluoroacetic
acid.
Example 1
N-((5-chlorothiophen-2-yl)methyl)-4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,-
2-dihydroquinazolin-3(4H)-yl)benzamide (4)
##STR00019## ##STR00020##
[0233] Step 1: 1a:
##STR00021##
[0235] To 3,4-difluorobenzoic acid (120.0 g, 0.7589 mol) in a 2-L
three-necked R. B. flask, was added concentrated H.sub.2SO.sub.4
(96.4%, 1.0 L). The mixture was mechanically stirred at room
temperature for 1.0 hr and then cooled to 0.degree. C. Fuming
HNO.sub.3 (HNO.sub.3 content >90%, 57.30 mL, 1.273 mmol, 1.677
equiv) was added dropwise over 30 minutes. The mixture was stirred
at room temperature for 16 h, slowly poured, in small portions,
over crushed ice (5 Kg) with stirring. 4,5-Difluoro-2-nitrobenzoic
acid precipitated out as a yellow solid. The slurry was stirred at
0.degree. C. for 0.5 hr and then the solid product was collected
via filtration through a Buckner funnel. The pale yellow solid la
was rinsed with chilled water, and then air dried; yield 120 g
(79%).
Step 2: 1b:
##STR00022##
[0237] To a solution of 4,5-difluoro-2-nitrobenzoic acid 1a (40.0
g, 196.947 mmol) in anhydrous MeOH (1.0 L), was added concentrated
H.sub.2SO.sub.4 (60.0 mL) and the clear mixture was refluxed for 2
days. Then methanol was removed in vacuo and the residue was
partitioned between EtOAc and water. The EtOAc extract was washed
with brine, dried over with anhydrous. Na.sub.2SO.sub.4, and
concentrated to yield the methyl 4,5-difluoro-2-nitrobenzoate 1b as
a pale yellow solid, 39.71 g (93%).
Step 3:
##STR00023##
[0239] To a solution of methyl 4,5-difluoro-2-nitrobenzoate 1b
(23.0 g, 106 mmol) in EtOAc (200 mL), was added 10% Pd/C (wet, 50%
water, 6.0 g), and the mixture was shaken on a Parr hydrogenator at
50 PSI for 3 hr. The reaction mixture was then filtered through a
celite pad and the filtrate was concentrated by rotary evaporation
to yield methyl 2-amino-4,5-difluorobenzoate 1c as a colorless
solid, 19.0g (96%).
Step 4:
[0240] To a solution of aniline 1c (4.8 g, 26 mmol) in DCM (26 mL),
was added diisopropylethyl amine (DIPEA) (4.9 mL, 28 mmol) and
methyl 4-isocyanatobenzoate (5.0 g, 28 mmol). The reaction mixture
was stirred for 2 days. A thick and off-white suspension was
formed. The solid was collected through filtration. It contained
the uncyclized urea (1d) and a small amount of the desired
quinazolinedione. To a portion of the solid (6.95 g, 19 mmol) in
dimethylsulfoxide (DMSO) (19 mL), was added methylamine (2M in THF,
19 mL, 38 mmol). The mixture was heated and stirred at 100.degree.
C. until all starting materials had been converted to the desired
product. The mixture was then cooled, diluted with water to a total
volume of 200 mL, and filtered. The solid was dried overnight by
aspiration and in vacuo to afford 1e. .sup.1H NMR (CDCl.sub.3, 400
MHz): .delta. 11.33 (s, 1H), 8.02 (d, 2H), 7.41 (d, 2H), 7.38 (d,
1H), 6.82 (s, 1H), 6.21 (d, 1H), 3.87 (s, 3H), 2.78 (d, 3H).
Step 5:
[0241] To a solution of methyl ester 1e from step 4 in 80 mL of
1,4-dioxane, was added 1 M aqueous lithium hydroxide (38 mL, 38
mmol). The mixture was stirred at room temperature overnight,
acidified with 3 M HCl until precipitation of the solid carboxylic
acid ceased, diluted with water to a total volume of 300 mL, and
filtered. The white solid was dried overnight by aspiration and
then in vacuo to afford if as a dense white powder in quantitative
yield. .sup.1H NMR (CDCl.sub.3, 400 MHz): .delta. 11.29 (s, 1H),
8.01 (d, 2H), 7.38 (m, 3H), 6.82 (s, 1H), 6.22 (d, 1H), 2.78 (s,
3H)
Step 6:
[0242] To a solution of 5-chlorothiophene-2-carboxaldehyde (1g) (3
mL, 28 mmol) in tetrahydrofuran (20 mL), was added sodium
borohydride (1.6 g, 42 mmol). The reaction mixture was stirred for
three hours at room temperature. The reaction was quenched with
saturated ammonium chloride followed by 1 M hydrochloric acid. This
mixture was then extracted with DCM and the organic layer was
concentrated in vacuo to afford alcohol (1h) as a colorless oil
.sup.1H NMR (CDCl.sub.3, 400 MHz): .delta. 6.78 (m, 2H), 4.71 (s,
2H).
Step 7:
[0243] To a solution of alcohol 1h (2.39 g, 16 mmol) in diethyl
ether (50 mL) at 0.degree. C., was added phosphorous tribromide
(1.5 mL, 16 mmol) slowly. The mixture was warmed to room
temperature and stirred for 2 days. The solution was diluted with
water, separated and the aqueous phase was extracted with diethyl
ether. The organic layer was dried over magnesium sulfate, filtered
and concentrated in vacuo to afford a 3:4 mixture of the desired
bromide 1i and the starting alcohol 1h. .sup.1H NMR (DMSO, 400
MHz): .delta. 7.09 (d, 1H), 6.97 (d, 1H), 4.95 (s, 2H). .sup.1H NMR
Minor (DMSO, 400 MHz): .delta.7.07 (d, 1H), 6.93 (d, 1H), 4.62 (s,
2H).
Step 8:
[0244] To the crude mixture from step 7 in DMF (10 mL), was added
sodium azide (2.0 g, 32 mmol). The mixture was stirred overnight,
diluted with water and extracted twice with DCM. The combined
organic layer was dried over magnesium sulfate, filtered and
concentrated. Flash column chromatography (0-20% DCM/hexanes)
afforded the desired alkyl azide (1j) which was used immediately
for the next step (1.02 g, 33%). .sup.1H NMR (DMSO, 400 MHz):
.delta. 7.03 (m, 2H), 4.59 (s, 2H).
Step 9:
[0245] To a solution of azide 1j (1.02 g, 5.9 mmol) in methanol (20
mL), was added tin chloride dihydrate (2.67 g, 11.8 mmol). The
mixture was stirred at room temperature until all of the azide had
been converted to the amine as determined by HPLC. The reaction
mixture was diluted with saturated sodium carbonate (10 mL) and
ethyl acetate was added. The organic layer was dried, filtered and
concentrated in vacuo to afford 1k as a light yellow oil. .sup.1H
NMR (DMSO, 400 MHz): .delta. 6.89 (d, 1H), 6.73 (d, 1H), 3.82 (s,
2H), 2.01 (br s, 2H).
Step 10:
[0246] To a solution of acid 1f (50 mg, 0.15 mmol) in DMF (2mL),
was added DIPEA (40 uL, 0.23 mmol) and
O-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (HATU) (104 mg, 0.28 mmol). After stirring for
10 min, the mixture was treated with amine 1k (41 mg, 0.28 mmol)
and stirred overnight. The mixture was diluted with water (15 mL)
and the resulting solid was isolated to afford the desired amide
(4) as light beige solid. MS found for
C.sub.21H.sub.16ClFN.sub.4O.sub.3S as (M+H).sup.- 459.1.
Example 2
N-((5-chlorothiophen-2-yl)methyl)-4-(6,7-difluoro-4-oxo-2H-benzo[e][1,3]ox-
azin-3(4H)-yl)benzamide (55)
##STR00024##
[0247] Step 1:
[0248] To a solution of trifluoride 2a (15 g, 96 mmol) in
N,N'-dimethylimidazolidinone (DMI) (200 mL), was added sodium
hydroxide (15 g, 382 mmol) slowly. The mixture was heated to
130.degree. C. and stirred until all the starting material was
consumed. The resulting thick beige semisolid was treated with
crushed ice (2 L), acidified with concentrated hydrochloric acid to
pH=2. The solid was collected by filtration and dried under vacuum
overnight to afford (2b) as a white solid (14.2 g). .sup.1H NMR
(DMSO, 400 MHz): .delta. 7.73 (dd, 1H), 7.08 (dd, 1H).
Step 2:
[0249] To a suspension of phenol 2b (5.5 g, 32 mmol) in pyridine
(10 mL), was added acetic anhydride (4.4 mL, 47 mmol) slowly. The
mixture was stirred for at room temperature for 1.5 hrs. The
reaction mixture was slowly poured into 3 M hydrochloric acid (120
mL) and extracted twice with ethyl acetate. The combined organic
layers were dried over magnesium sulfate, filtered and concentrated
to afford acetate 2c as a white solid (5.12 g, 72%). .sup.1H NMR
(DMSO, 400 MHz): .delta. 7.90 (dd, 1H), 7.49 (dd, 1H), 2.22 (s,
3H).
Step 3:
[0250] To a solution of acid 2c (1.16 g, 5.4 mmol) in DCM (20 mL),
was added oxalyl chloride (0.70 mL, 8.1 mmol) followed by several
drops of DMF. After gas evolution ceased, the reaction was checked
by HPLC and determined to be complete. The mixture was concentrated
and used immediately for the next step.
Step 4:
[0251] To a solution of methyl 4-aminobenzoate (0.90 g, 6.0 mmol)
and DIPEA (1.4 mL, 3 eq.) in DCM (10 mL), was added crude acid
chloride from step 3 in DCM (10 mL) dropwise. After the reaction
was found to be complete by HPLC, the mixture was diluted with
water and extracted once with DCM. The combined organic phases were
dried over magnesium sulfate, filtered, then concentrate to afford
a solid (1.34 g, 71% yield) as a mixture of the acetate 2e and the
free phenol 2f.
Step 5:
[0252] To a solution of the crude product from the step 4 ((1.34 g,
3.8 mmol) in methanol (10 mL) and dioxane (10 mL), was added
potassium carbonate (0.53 g, 3.8 mmol). The reaction mixture was
stirred at room temperature for 90 min and then acidified with 1 M
hydrochloric acid. Upon sitting overnight, a precipitate formed
which was filtered and dried overnight by aspiration to afford
phenol 2f as a white solid (0.86 g, 74%). .sup.1H NMR (DMSO, 400
MHz): .delta. 11.86 (s, 1H), 10.57 (s, 1H), 7.96 (d, 2H), 7.94 (m,
1H), 7.83 (d, 2H), 7.00 (dd, 1H), 3.82 (s, 3H).
Step 6:
[0253] Amide 2f (0.86 g, 2.8 mmol) was combined with
paraformaldehyde (0.42 g, 14 mmol) and toluene sulfonic acid (0.21
g, 1.12 mmol), the mixture was then diluted with 30 mL toluene and
heated to reflux for 2 hours under a Dean-Stark trap previously
filled with toluene. The reaction mixture was cooled to rt and
partitioned with sodium bicarbonate and ethyl acetate. The aqueous
phase was extracted again with ethyl acetate. The combined organic
phases were dried over magnesium sulfate, filtered and concentrated
to afford 2g as a light beige solid (0.92 g, quant). .sup.1H NMR
(DMSO, 400 MHz): .delta. 8.02 (d, 2H), 7.91 (dd, 1H), 7.50 (d, 2H),
7.46 (dd, 1H), 5.82 (s, 2H), 3.88 (s, 3H).
Step 7:
[0254] To a solution of difluoride 2g (0.92 g, 2.9 mmol) in DMSO
(10 mL), was added methylamine (2 M in THF, 4.4 mL, 8.8 mmol). The
reaction mixture was heated at 120.degree. C. in a sealable tube
for three hours. It was then cooled and diluted with water (100
mL). The resulting white precipitate was isolated by filtration and
dried to afford 2h (0.81 g, 86%) as a white solid. .sup.1H NMR
(DMSO, 400 MHz): .delta. 7.96 (d, 2H), 7.49 (d, 2H), 7.36 (d, 1H),
6.72 (s, 1H), 6.25 (d, 1H), 5.68 (s, 2H), 3.84 (s, 3H), 2.76 (s,
3H).
Step 8:
[0255] To a solution of methyl ester 2h (200 mg, 0.60 mmol) in
dioxane (5 mL), was added 1 M lithium hydroxide (1.2 mL, 1.2 mmol).
The mixture was stirred until all ester had been consumed (HPLC).
The reaction was concentrated in vacuo, diluted with 1 mL 1M HCl
and 3 mL of water. The resulting white solid was filtered and dried
overnight to afford 2i (178 mg, 94%) as a light beige powder.
.sup.1H NMR (DMSO, 400 MHz): .delta. 7.97 (d, 2H), 7.48 (d, 2H),
7.36 (d, 1H), 7.10 (br s, 1H), 6.71 (s, 1H), 6.24 (d, 1H), 5.68 (s,
2H), 2.76 (s, 3H).
Step 9:
[0256] To a solution of carboxaldehyde 1g (168 uL, 1.58 mmol) and
glycine ethyl ester hydrochloride 2j (200 mg, 1.44 mmol) in
dichloroethane (10 mL), was added triethyl amine (TEA) (270 uL,
1.58 mmol), followed by sodium triacetoxyborohydride (0.46 g, 2.16
mmol). The mixture was stirred at room temperature overnight. The
mixture was diluted with aqueous sodium carbonate/water/ethyl
acetate. The layers separated and the aqueous phase was extracted
with ethyl acetate and the combined organic phases was dried over
magnesium sulfate, filtered, and concentrated in vacuo. The crude
residue was purified by silica gel chromatography (1-10% ethyl
acetate/DCM) to afford the desired amine 2k (73 mg, 22%) as a light
yellow oil (.sup.1H NMR (DMSO, 400 MHz): .delta. 6.93 (d, 1H), 6.81
(d, 1H), 4.10 (q, 2H), 3.85 (s, 2H), 1.18 (t, 3H).
Step 10:
[0257] Compound 2L was synthesized in a manner similar to Example
1, Step 10. MS found for C.sub.25H.sub.23ClFN.sub.3O.sub.5S as
(M+H).sup.+ 532.2.
Step 11:
[0258] Compound 55 was synthesized in a manner similar to Example
1, Step 5. MS found for C.sub.23H.sub.19ClFN.sub.3O.sub.5S as
(M+H).sup.+ 504.1.
Example 3-5
2-(N-((5-chlorothiophen-2-yl)methyl)-4-(6-fluoro-7-(methylamino)-2,4-dioxo-
-1,2-dihydroquinazolin-3(4H)-yl)benzamido)acetic acid (Compound
6)
##STR00025##
[0259]
3-(N-((5-chlorothiophen-2-yl)methyl)-4-(6-fluoro-7-(methylamino)-2,-
4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamido)propanoic acid
(Compound 40)
##STR00026##
[0260]
3-(N-((5-chlorothiophen-2-yl)methyl)-4-(6-fluoro-7-(methylamino)-4--
oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamido)propanoic acid
(Compound 86)
##STR00027##
[0262] Compounds 6, 40 and 86 were made by the procedure similar to
that described for Example 2. MS found for Compound 6 as
(M+H).sup.+: 517.0. MS found for Compound 40 as (M+H).sup.+: 531.0.
MS found for Compound 86 as (M+H).sup.+: 518.0.
Example 6
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dih-
ydroquinazolin-3(4H)-yl)benzamido)acetic acid (5)
##STR00028##
[0263] Step 1:
[0264] To a suspension of amino acid 3a (10 g, 63 mmol) in a
mixture of dioxane (300 mL) and methanol (150 mL), was added
trimethylsilyldiazomethane (2 M in diethyl ether, 48 mL, 96 mmol).
The mixture was stirred at room temperature until all starting acid
had been converted to the methyl ester 3b. The mixture was
concentrated to thick oil which was used immediately for the next
step.
Step 2:
[0265] To a solution of 3b in acetic acid (120 mL) at 0.degree. C.,
was added sulfuryl chloride (4.8 mL, 60 mmol) in three portions.
More sulfuryl chloride (3 mL) was added slowly until all of the
starting thiophene had been consumed. The crude mixture was
concentrated in vacuo, then partitioned with aqueous sodium
bicarbonate (carefully) and DCM. The organic layer was
concentration and the crude residue was purified by silica gel
chromatography (0-5% ethyl acetate/DCM) to afford 3c as a light
yellow oil. The oil was dissolved in diethyl ether (100 mL) and
acidified with 4 M hydrochloric acid in dioxane with vigorous
stirring. The solid was then filtered to afford 3c as a beige HCl
salt (6.24 g, 51%). MS found for C.sub.7H.sub.8ClNO.sub.2S as
(M+H).sup.+ 206.0, 208.0.
Step 3:
[0266] Compound 3d (18) was synthesized in a manner similar to
Example 1, Step 10. .sup.1H NMR (DMSO, 400 MHz): .delta. 11.31 (s,
1H), 9.58 (d, 1H), 7.95 (d, 2H), 7.38 (m, 3H), 7.03 (d, 1H), 6.82
(s, 1H), 6.24 (d, 1H), 5.84 (d, 1H), 3.70 (s, 3H), 2.73 (d,
3H).
Step 4:
[0267] Compound 5 was synthesized by the procedure similar to that
described for Example 1, Step 5. MS found for
C.sub.22H.sub.16ClFN.sub.4O.sub.5S as (M+H).sup.+ 503.0.
Example 7-8
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e]-
[1,3]oxazin-3(4H)-yl)benzamido)acetic acid (Compound 56)
##STR00029##
[0268]
3-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-b-
enzo[e][1,3]oxazin-3(4H)-yl)benzamido)propanoic acid (Compound
57)
##STR00030##
[0270] Compounds 56 and 57 were synthesized by the procedure
similar to that described for Example 6. MS found for Compound 56
as (M+H).sup.+: 490.1. MS found for Compound 57 as (M+H).sup.+:
504.1.
Example 9
3-(5-chlorothiophen-2-yl)-3-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dih-
ydroquinazolin-3(4H)-yl)benzamido)propanoic acid (27)
##STR00031##
[0271] Step 1:
[0272] To a solution of thiophene carboxaldehyde 4a (3 mL, 30 mmol)
in 1,4-dioxane (20 mL), was added malonic acid (3.43 g, 33 mmol)
and ammonium acetate (4.6 g, 60 mmol). The mixture was heated to
reflux overnight. It was cooled to rt and filtered. Upon sitting
overnight, a precipitate formed which was isolated through
filtration and dried in vacuo to afford 4b as a white solid (2.13
g, 34%). NMR (DMSO, 400 MHz): .delta. 7.54 (d, 1H), 7.28 (d, 1H),
7.05 (dd, 1H), 4.81 (dd, 1H), 3.04 (dd, 1H), 2.90 (dd, 1H).
Step 2:
[0273] To a solution of amino acid 4b (1.38 g, 8.1 mmol) DCM (10
mL) and methanol (10 mL), was added trimethylsilyldiazomethane (2 M
in diethyl ether, 6 mL, 12 mmol). The reaction mixture was stirred
at rt for 90 min and concentrated in vacuo. The residue was
purified by silica gel chromatography (0-70% ethyl acetate/DCM) to
afford the desired amino ester 4c as a colorless oil. NMR (DMSO,
400 MHz): .delta. 7.33 (d, 1H), 6.91 (m, 2H), 4.44 (dd, 1H), 3.56
(s, 3H), 2.68 (m, 2H).
Step 3:
[0274] Compound 4d was synthesized in a manner similar to Example
6, Step 2. MS found for C.sub.8H.sub.8ClO.sub.2S as
(M-NH.sub.2).sup.+ 203.0, 205.0.
Step 4:
[0275] Compound 4e was synthesized in a manner similar to Example
1, Step 10. NMR (DMSO, 400 MHz): .delta. 9.10 (d, 1H), 7.98 (d,
1H), 7.88 (d, 2H), 7.36 (m, 3H), 6.93 (d, 1H), 6.77 (s, 1H), 6.25
(d, 1H), 5.58 (dd, 1H), 3.58 (s, 3H), 3.05 (d, 2H), 2.77 (d,
3H).
Step 5:
[0276] Compound 27 was synthesized in a manner similar to Example
1, Step 5. MS found for C.sub.23H.sub.18ClFN.sub.4O.sub.5S as
(M-H).sup.- 515.0, 517.0.
Example 10
3-(5-chlorothiophen-2-yl)-3-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e]-
[1,3]oxazin-3(4H)-yl)benzamido)propanoic acid (Compound 79)
##STR00032##
[0278] Compound 79 was synthesized in a manner similar to Example
9. MS found for Compound 79 as (M+H).sup.+: 504.0.
Example 11
N-((5-chlorothiophen-2-yl)(1H-tetrazol-5y1)methyl)-4-(6-fluoro-7-(methylam-
ino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide (Compound
7)
##STR00033##
[0279] Step 1:
[0280] Aldehyde 1g (3 mL, 28 mmol) was mixed with
trimethylsilylcyanide (4.5 mL, 34 mmol) and ca. 50 mg of ZnI.sub.2.
The resulting brown solution was stirred for 30 min at rt, then
treated with ammonia (7 M in methanol, 20 mL, 140 mmol). The
mixture was heated to 50.degree. C. for 90 min. in a sealable tube.
Ethyl acetate and water were added. The layers separated and the
aqueous phase was extracted with ethyl acetate. The combined
organic phases were concentrated to afford 5a (2.17 g, 45%) as a
light beige solid. (DMSO, 400 MHz): .delta. 7.00 (d, 1H), 6.97 (d,
1H), 5.24 (s, 1H), 3.10 (s, 2H).
Step 2:
[0281] Compound 5b was synthesized in a manner similar to Example
1, Step 10. MS found for C.sub.22H.sub.15ClFN.sub.5O.sub.3S as
(M+H).sup.+ 484.1.
Step 3:
[0282] To a solution of the nitrile 5b (65 mg, 0.14 mmol) in a 1:1
mixture of isopropanol and water (5 mL), was added sodium azide (18
mg, 0.28 mmol) and zinc bromide (16 mg, 0.07 mmol). The mixture was
stirred at 80.degree. C. until complete consumption of the starting
material. The mixture was cooled to rt, concentrated in vacuo, and
purified immediately by prep HPLC to afford compound 7 as a white
solid. MS found for C.sub.22H.sub.16ClF.sub.8O.sub.3S as
(M-H).sup.- 525.0, 527.0.
Examples 12-24
N-((5-chlorothiophen-2-yl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(methyla-
mino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamide (Compound
54)
##STR00034##
[0283]
N-((3-chlorophenyl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(methyla-
mino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide (Compound
11)
##STR00035##
[0284]
N-((4-chlorophenyl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(methyla-
mino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide (Compound
12)
##STR00036##
[0285]
4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)--
N-((5-methylthiophen-2-yl)(1H-tetrazol-5-yl)methyl)benzamide
(Compound 16)
##STR00037##
[0286]
N-((1H-tetrazol-5-yl)(m-tolyl)methyl)-4-(6-fluoro-7-(methylamino)-2-
,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide (Compound 22)
##STR00038##
[0287]
4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-y-
l)-N-((3-methoxyphenyl)(1H-tetrazol-5-yl)methyl)benzamide (Compound
23)
##STR00039##
[0288]
4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-y-
l)-N-((3-fluorophenyl)(1H-tetrazol-5-yl)methyl)benzamide (Compound
24)
##STR00040##
[0289]
N-((2-chlorophenyl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(methyla-
mino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide (Compound
28)
##STR00041##
[0290]
N-((3,4-dichlorophenyl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(met-
hylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide
(Compound 29)
##STR00042##
[0291]
N-((3,5-dichlorophenyl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(met-
hylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide
(Compound 30)
##STR00043##
[0292]
N-((1H-tetrazol-5-yl)(3-(trifluoromethoxy)phenyl)methyl)-4-(6-fluor-
o-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide
(Compound 31)
##STR00044##
[0293]
N-((1H-tetrazol-5-yl)(3-(trifluoromethyl)phenyl)methyl)-4-(6-fluoro-
-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide
(Compound 34)
##STR00045##
[0294]
4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)--
N-((5-methylthiophen-2-yl)(1H-tetrazol-5-yl)methyl)benzamide
(Compound 61)
##STR00046##
[0296]
N-((5-chlorothiophen-2-yl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(-
methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamide (54)
and compounds 11, 12, 16, 22, 23, 24, 28, 29, 30, 31, 34 and 61
were synthesized by the procedure similar to that described for
Example 11. Compound 54. (DMSO, 400 MHz): .delta. 9.76 (s, 1H),
7.94 (d, 2H), 7.47 (d, 2H), 7.35 (d, 1H), 7.04 (s, 2H), 6.88 (d,
1H), 6.71 (s, 1H), 6.25 (d, 1H), 5.64 (s, 2H), 2.74 (s, 3H). MS
found for Compound 54 as (M+H).sup.+: 514.0. MS found for Compound
11 as (M+H).sup.+: 521.3. MS found for Compound 12 as (M+H).sup.+:
521.3. MS found for Compound 16 as (M+H).sup.+: 507.0. MS found for
Compound 22 as (M+H).sup.+: 501.0. MS found for Compound 23 as
(M+H).sup.+: 517.0. MS found for Compound 24 as (M+H).sup.+: 505.0.
MS found for Compound 28 as (M+H).sup.+: 521.0. MS found for
Compound 29 as (M+H).sup.+: 556.0. MS found for Compound 30 as
(M+H).sup.+: 556.0. MS found for Compound 31 as (M+H).sup.+: 571.0.
MS found for Compound 34 as (M+H).sup.+: 555.0. MS found for
Compound 61 as (M+H).sup.+: 492.3.
Example 25
N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)methyl)--
4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benza-
mide (21)
##STR00047## ##STR00048##
[0297] Step 1:
[0298] To a solution of nitrile 5b (100 mg, 0.21 mmol) in DMSO (1
mL), was added hydroxylamine hydrochloride (73 mg, 1.05 mmol) and
DIPEA (183 uL, 1.05 mmol). The reaction mixture was heated to
70.degree. C. for 2 hrs. The reaction mixture was diluted with
water (ca. 10 mL). The resulting solid was isolated through
filtration to afford 6a as a light beige solid in quantitative
yield. (DMSO, 400 MHz): .delta. 9.08 (d, 1H), 7.82 (d, 2H), 7.37
(m, 3H), 6.97 (d, 1H), 6.91 (d, 1H), 6.84 (s, 1H), 6.27 (d, 1H),
5.88 (d, 1H), 5.73 (s, 1H), 2.77 (d, 3H).
Step 2:
[0299] To a solution of hydroxamidine 6a (50 mg, 0.10 mmol) in DMF
(2 mL), was added pyridine (9 uL, 0.11 mmol), followed by ethyl
chloroformate (10 uL, 0.10 mmol). The mixture was stirred at rt
overnight. Water (10 mL) was added and the solid was isolated
through filtration to afford 6b in quantitative yield. (DMSO, 400
MHz): .delta. 11.28 (s, 1H), 9.13 (d, 1H), 7.93 (d, 2H), 7.42 (d,
2H), 7.35 (s, 1H), 7.00 (s, 2H), 6.83 (s, 1H), 6.77 (s, 2H), 6.25
(d, 1H), 5.93 (d, 1H), 3.97 (q, 2H), 2.79 (d, 3H), 1.24 (t,
3H).
Step 3:
[0300] To a solution of carbonate 6b (465 mg, 0.90 mmol) in DMF (5
mL), was added cesium carbonate (350 mg, 1.1 mmol). The mixture was
stirred at rt overnight. The reaction mixture was diluted with
water and 1 M HCl to a total volume of 40 mL and a pH of
approximately 3. The solid was filtered and washed with water, then
purified by prep HPLC to afford compound 21 (107 mg, 22%). (DMSO,
400 MHz): .delta. 11.30 (s, 1H), 9.29 (d, 1H), 7.91 (d, 2H), 7.34
(m, 3H), 6.80 (s, 2H), 6.26 (d, 1H), 6.13 (d, 1H), 2.74 (d, 3H). MS
found for Compound 21 as (M+H).sup.+:543.0.
Example 26
N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-thiadiazol-3-yl)methyl--
4-6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzam-
ide (Compound 25)
##STR00049##
[0302] To a solution of 6a (100 mg, 0.19 mmol) in tetrahydrofuran
(2 mL), was added thiocarbonyldiimidazole (37 mg, 0.21 mmol). The
reaction mixture was stirred at rt for 30 min. Silica gel (1 g) in
20 mL of a 5:1 mixture of chloroform and methanol was added to the
reaction and the suspension was stirred at rt overnight. The
mixture was concentrated and purified by prep HPLC to afford 25 as
a white solid (9 mg, 10%). MS found for
C.sub.23H.sub.16ClFN.sub.6O.sub.4S.sub.2 as (M-H).sup.- 557.0,
559.0.
Example 27
N-((5-chlorothiophen-2-yl)(5-thioxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)methy-
l)-4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)be-
nzamide (Compound 32)
##STR00050##
[0304] To a solution of 6a (100 mg, 0.19 mmol) in DMF (3 mL), was
added thiocarbonyldiimidazole (52 mg, 0.29 mmol) and DBU (113 uL,
0.76 mmol). The mixture was stirred at rt for 4-5 hours, diluted
with water and purified by prep HPLC to afford 32 as a white solid.
MS found for C.sub.23H.sub.16ClFN.sub.6O.sub.4S.sub.2 as
(M-H).sup.- 556.7, 558.8.
Examples 28
N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)methyl)--
4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamid-
e (Compound 71)
##STR00051##
[0306] Compound 71 was made by the procedure similar to that
described for Example 25. Title compound: MS found for
C.sub.23H.sub.18ClFN.sub.4O.sub.5S as (M+H).sup.+530.0.
Example 29
N-((5-chlorothiophen-2-yl)(5-thioxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)methy-
l)-4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benza-
mide (Compound 75)
##STR00052##
[0308] Compound 75 was synthesized by the procedure similar to that
described for Example 27. MS found for Compound 75 as (M+H).sup.+:
545.8.
Example 30
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dih-
ydroquinazolin-3(4H)-yl)-3-methoxybenzamido)acetic acid (51)
##STR00053## ##STR00054##
[0309] Step 1:
[0310] To difluoroanthranalide 1c (5.69 g, 30 mmol), was added
phosgene in toluene ((1.9 M, 160 mL, 300 mmol). The reaction
mixture was stirred at rt overnight, concentrated in vacuo,
evaporated from toluene (20 mL), and dried under vacuum to afford a
light beige solid 8a. The material was found to give the desired
urea when treated with ammonia but was not otherwise
characterized.
Step 2:
[0311] Anhydrous HCl was made by addition of thionyl chloride (1.46
mL, 20.4 mmol) and methanol (10 mL) at 0.degree. C. To this was
added 3-methoxy-4-nitrobenzoic acid 8b (2.00 g, 10.2 mmol). The
mixture was stirred at room temperature overnight, concentrated,
diluted with aqueous sodium bicarbonate, and extracted with
dichloromethane. The organic layer was concentrated in vacuo to
afford methyl ester 8c in quantitative yield. H.sup.1 NMR (400 MHz,
CDCl.sub.3) .delta. 7.82 (d, 1H), 7.75 (s, 1H), 7.68 (d, 1H), 4.02
(s, 3H), 3.99 (s, 3H).
Step 3:
[0312] To solution of 8c in ethyl acetate (20 mL), was added 10%
Pd/C (Degussa, 200 mg). The reaction mixture was stirred under an
atmosphere of hydrogen overnight, filtered through celite and
concentrated to give the aniline 8d as a white solid (1.54 g, 83%
for 2 steps). H.sup.1 NMR (400 MHz, DMSO-d.sub.6) .delta. 3.73 (s,
3H), 3.78 (s, 3H), 5.59 (s, 2H), 6.60 (d, 1H), 7.28 (s, 1H), 7.35
(dd, 1H).
Step 4:
[0313] To a solution of aniline 8d (200 mg, 1.1 mmol) in DCM (10
mL), was added DIPEA (290 uL, 1.66 mmo), followed by carbamoyl
chloride 8a (0.35 g, 1.66 mmol). The mixture was stirred overnight,
concentrated in vacuo, and the crude product (urea and a small
amount of 8e) was used for the next step without further
purification.
Step 5:
[0314] To a solution of crude product from step 4 in DMSO (5 mL),
was added methyl amine (2M in THF, 1.7 mL 3.3 mmol). The mixture
was heated to 100.degree. C. in a sealable tube overnight. The
reaction mixture was diluted with water and the solid was isolated
through filtration to afford 8e as a light beige solid (330 mg, 80%
for 2 steps). MS found for C.sub.18F.sub.16FN.sub.3O.sub.5 as
(M+H).sup.+ 374.2.
Step 6:
[0315] Compound 8f was synthesized in a manner similar to Example
1, Step 5.
Step 7:
[0316] Compound 8g was prepared using a procedure similar to
Example 1, step 10 using 8f and 3c. (DMSO, 400 MHz): .delta. 7.68
(d, 1H), 7.60 (d, 1H), 7.45 (d, 1H), 7.33 (d, 1H), 6.93 (d, 1H),
6.93 (d, 1H), 6.80 (d, 1H), 6.31 (d, 1H), 5.65 (s, 1H), 3.88 (s,
3H), 2.93 (s, 3H).
Step 8:
[0317] Compound 51 was synthesized using a procedure similar to
Example 1, step 5. MS found for Compound 51 as
(M+H).sup.+:533.0.
Example 31
2-(3-chlorophenyl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroqui-
nazolin-3(4H)-yl)benzamido)acetic acid 15
##STR00055##
[0318] Step 1:
[0319] Aminonitrile 7a (prepared from 3-chlorobenzaldehyde using
similar procedure described for 5a) (0.76 g, 3.8 mmol) was
dissolved in 6 M hydrochloric acid (15 mL). The mixture was
refluxed for 2 hrs, cooled to rt, concentrated and used immediately
for the next step.
Step 2:
[0320] Compound 7c was synthesized in a manner similar to Example
30, Step 2. The crude product was used immediately for the next
step.
Step 3:
[0321] Compound 7d was synthesized in a manner similar to Example
1, Step 10.
Step 4:
[0322] Compound 15 was synthesized in a manner similar to Example
1, Step 5. (DMSO, 400 MHz): .delta. 11.27 (s, 1H), 9.17 (dd, 1H),
7.92 (d, 2H), 7.53 (m, 2H), 7.38 (m, 4H), 6.80 (s, 1H), 6.23 (d,
1H), 5.64 (dd, 1H), 2.77 (d, 3H). MS found for Compound 15 as
(M+H).sup.+: 497.0.
Examples 32-38
2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)be-
nzamido)-2-(5-methylthiophen-2-yl)acetic acid (Compound 19)
##STR00056##
[0323]
2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-y-
l)benzamido)-2-phenylacetic acid (Compound 58)
##STR00057##
[0324]
(S)-2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4-
H)-yl)benzamido)-3-phenylpropanoic acid (Compound 60)
##STR00058##
[0325]
2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-y-
l)benzamido)-2-(thiophen-2-yl)acetic acid (Compound 63)
##STR00059##
[0326]
2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-y-
l)benzamido)-3-(thiophen-2-yl)propanoic acid (Compound 64)
##STR00060##
[0327]
(S)-2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4-
H)-yl)benzamido)-2-phenylacetic acid (Compound 65)
##STR00061##
[0328]
2-(3-chlorophenyl)-2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e]-
[1,3]oxazin-3(4H)-yl)benzamido)acetic acid (Compound 81)
##STR00062##
[0330] Compounds 19, 58, 60, 63, 64, 65, and 81 were synthesized by
the procedure similar to that described for Example 31 from the
corresponding commercially available amino acid after
esterification. MS found for Compound 19 as (M+H).sup.+: 483.0. MS
found for Compound 58 as (M+H).sup.+: 450.1. MS found for Compound
60 as (M+H).sup.+: 464.2. MS found for Compound 63 as (M+H).sup.+:
456.0. MS found for Compound 64 as (M+H).sup.+: 470.2. MS found for
Compound 65 as (M+H).sup.+: 448.2. MS found for Compound 81 as
(M+H).sup.+: 484.0.
Example 39
2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benza-
mido)-3-phenylpropanoic acid (Compound 59)
##STR00063##
[0332] The titled compound was made by the procedure similar to
that described for the Examples above from the corresponding
commercially available amino acid. (DMSO, 400 MHz): .delta. 8.71
(d, 1H), 7.82 (d, 2H), 7.40 (d, 2H), 7.13-7.38 (m, 6H), 6.72 (s,
1H), 6.22 (d, 1H), 5.63 (s, 2H), 4.60 (m, 1H), 3.18 (dd, 1H), 3.07
(dd, 1H), 2.78 (s, 3H).
Example 40
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dih-
ydroquinazolin-3(4H)-yl)-2-methylbenzamido)acetic acid (Compound
41)
##STR00064##
##STR00065##
[0333] Step 1:
[0334] Compound 9b was synthesized in a manner similar to Example
30, Step 2. H.sup.1 NMR (400 MHz, CDCl.sub.3) .delta. 7.78 (d, 1H),
7.42 (s, 1H), 7.37 (d, 1H), 3.87 (s, 3H), 2.58 (s, 3H).
Step 2:
[0335] To a solution of aryl bromide 9b (1.00 g, 4.4 mmol) in THF
(20 mL), was added t-butyl carbamate (0.61 g, 5.2 mmol), cesium
carbonate (2.87 g, 8.8 mmol), Xantphos (8.38 g, 0.66 mmol). The
solution was degassed with argon. To this solution, was added
Pd(dba).sub.2Cl.sub.2 and the resulting mixture was refluxed
overnight, diluted with water and extracted twice with DCM. The
combined organic phases was concentrated and purified by silica gel
chromatography (DCM) to afford carbamate 9c (1.08 g, 93%). H.sup.1
NMR (400 MHz, DMSO-d.sub.6) .delta. 9.62 (s, 1H), 7.77 (d, 1H),
7.38 (m, 2H), 3.73 (s, 3H), 2.43 (s, 3H), 1.43 (s, 9H).
Step 3:
[0336] Boc protected amine 9c (1.04 g, 3.9 mmol) was dissolved in 4
M hydrochloric acid in dioxane (10 mL) and stirred at rt for 3
hours. The reaction was diluted slowly with a small amount of
diethyl ether until a filterable solid formed. The solid was
isolated through filtration and washed with diethyl ether to afford
9d as a light beige solid (HCl salt) (482 mg, 61%). H.sup.1 NMR
(400 MHz, DMSO-d.sub.6) .delta. 8.01 (d, 1H), 7.25 (m, 2H), 3.88
(s, 3H), 2.61 (s, 3H).
Step 4:
[0337] Compound 41 was prepared using a procedure similar to
Example 30. MS found for C.sub.23H.sub.18ClFN.sub.4O.sub.5S as
(M-H).sup.- 515.0, 517.0.
Example 41
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dih-
ydroquinazolin-3(4H)-yl)-2-methoxybenzamido)acetic acid (Compound
42)
##STR00066##
[0339] Compound 42 was synthesized using procedures similar to
steps 1-3 of Example 40 and step 4 of Example 30 starting from
methyl 4-amino-2-methoxybenzoate. MS found for Compound 42 as
(M+H).sup.+: 533.0.
Example 42
2-(2-chloro-4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(-
4H)-yl)benzamido)-2-(5-chlorothiophen-2-yl)acetic acid (Compound
43)
##STR00067##
[0341] Compound 43 was synthesized starting from
4-bromo-2-chlorobenzoic acid using a procedure similar to Example
30. MS found for Compound 43 as (M+H).sup.+: 537.0.
Example 43
2-(5-chlorothiophen-2-yl)-2-(2-fluoro-4-(6-fluoro-7-(methylamino)-2,4-diox-
o-1,2-dihydroquinazolin-3(4H)-yl)benzamido)acetic acid (Compound
44)
##STR00068##
[0343] Compound 44 was synthesized starting from
4-bromo-2-fluorobenzoic acid using procedures similar to steps 1-3
of Example 40 and step 4 of Example 30. MS found for Compound 44 as
(M+H).sup.+: 521.0.
Example 44
N-((5-chlorothiophen-2-yl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-methylam-
ino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)-3-methoxybenzamide
(Compound 52)
##STR00069##
[0345] Compound 52 was prepared from the intermediate 8f in Example
30 and 5a in Example 11, using a procedure similar to that
described in Example 11. MS found for Compound 52 as (M+H).sup.+:
557.0.
Example 45
N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)methyl)--
4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)-3-me-
thoxybenzamide (Compound 53)
##STR00070##
[0347] Compound 53 was prepared from the intermediate 8f in Example
30 using a procedure similar to that described in Example 25.
(DMSO, 400 MHz): .delta. 9.26 (d, 1H), 7.55 (s, 1H), 7.48 (d, 1H),
7.30 (d, 1H), 7.21 (d, 1H), 6.86 (d, 2H), 6.73 (s, 1H), 6.17 (d,
1H), 6.08 (d, 1H), 3.70 (s, 3H), 2.72 (d, 3H). MS found for
Compound 53 as (M+H).sup.+: 573.0.
Example 46
2-(5-chlorothiophen-2-yl)-2-(3-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dih-
ydroquinazolin-3(4H)-yl)benzamido)acetic acid (Compound 111)
##STR00071##
[0349] Compound 111 using procedures similar to steps 1-3 of
Example 40 and step 4 of Example 30. (DMSO, 400 MHz): .delta. 8.28
(d, 1H), 7.73 (d, 1H), 7.53 (s, 1H), 7.47 (t, 1H), 7.25 (d, 1H),
7.19 (d, 1H), 6.80 (d, 1H), 6.68 (d, 1H), 6.05 (d, 1H), 6.00 (s,
1H), 5.04 (d, 1H), 2.77 (d, 3H). MS found for Compound 111 as
(M+H).sup.+: 503.0.
Example 47
##STR00072## ##STR00073##
[0350] Step 1:
[0351] To a solution of (R)-phenylglycinol 9a (5.0 g, 34 mmol) and
aldehyde 5a (5.6 g, 41 mmol) in methanol (70 mL) and water (20 mL),
was added sodium bisulfite (4.3 g, 41 mmol) and potassium cyanide
(2.7 g, 41 mmol). The resulting suspension was stirred until only a
small amount of the imine remained. The mixture was concentrated in
vacuo. DCM and water were added. The aqueous layer was extracted
with DCM and the organic layer was concentrated in vacuo. The crude
material was purified by silica gel chromatography (DCM) to afford
9b as a yellow oil (4.62 g, 46%) consisting of a 2:1 ratio of
diastereomers. (DMSO, 400 MHz): .delta. 7.37 (m, 5H), 7.28 (m, 3H),
7.0 (m, 2.5H), 5.04 (t, 1H), 4.88 (t, 0.5H), 4.77 (d, 1H), 3.93 (m,
1H), 3.74 (m, 1.5H), 3.43 (m, 3H).
Step 2:
[0352] Nitrile 9b was converted to hydroxamidine 9c using a
procedure similar to that described in Example 6, Step 1. The crude
material was used for the next step without purification. MS found:
(M+H).sup.+ 326.3, 328.0.
Step 3:
[0353] Hydroxamidine 9c was converted to carbonate 9d using a
procedure similar to that described in Example 6, Step 2. MS found:
(M+H).sup.+ 398.0, 400.0.
Step 4:
[0354] Carbonate 9d was converted to oxadiazolone 9f using a
procedure similar to that described in Example 6, Step 3. At this
stage the two diastereomers can be separated by preparative HPLC.
MS found: (M+H).sup.+ 352.0, 354.0.
Step 5:
[0355] To a solution of oxadiazolone 9f (30 mg, 0.085 mmol) in DCM
(6 mL) and methanol (3 mL) at 0.degree. C., was added lead
tetraacetate (38 mg, 0.085 mmol). The mixture was stirred for 5
min. The reaction mixture was concentrated and purified by prep
HPLC to afford amine 9g as a white solid which was immediately used
for the next step. MS found: (M-H).sup.- 230.2, 232.0.
Step 6:
[0356] Compound 45 was prepared using a procedure similar to that
described in Example 1, Step 10. The enantiomeric purity of the
material was measured by chiral HPLC using an (R,R)-ULMO column (25
cm, .times.4.6 mm, 5 um) from Regis Technologies eluting with 75/25
hexane/ethanol with 25 mM ammonium acetate and 1% triethyl amine
and found to be greater than 90% ee. MS found for Compound 45 as
(M+H).sup.+: 542.8.
Example 48
(R)--N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)met-
hyl-4-6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)be-
nzamide (Compound 46)
##STR00074##
[0358] Compound 46 was prepared using a procedure similar to that
described for Example 47, starting with (S)-phenylglycinol. MS
found: (M-H).sup.- 541.0, 543.0. Chiral purity using above
conditions was found to be greater than 90% ee.
Example 49
2-(5-chlorothiophen-2-yl)-2-(4-(7-fluoro-6-(methylamino)-2,4-dioxo-1,2-dih-
ydroquinazolin-3(4H)-yl)benzamido)acetic acid 48
##STR00075## ##STR00076##
[0359] Step 1:
[0360] To a solution of difluoroarene 1a (3.0 g, 14 mmol) in DMF (5
mL), was added methyl amine (2 M in THF, 24 mL, 48 mmol). The
mixture was stirred at rt until the completion of the reaction. The
mixture was diluted with water and extracted twice with ethyl
acetate. The combined organic layers were dried over magnesium
sulfate, filtered, and concentrated in vacuo to afford 10a as a red
oil which was used immediately for the next step. (DMSO, 400 MHz):
.delta. 7.91 (d, 1H), 7.39 (s, 1H), 6.79 (d, 1H), 3.83 (s, 3H),
2.83 (d, 3H).
Step 2:
[0361] To a solution of 10a in acetonitrile (20 mL), was added
t-butyl pyrocarbonate (3.3 g, 15 mmol) and
N,N-dimethylaminopyridine (0.68 g, 5.6 mmol). The mixture was
stirred for a few minutes, diluted with water and extracted twice
with DCM. The combined organic layer was dried over magnesium
sulfate, filtered, and concentrated to afford carbamate, 10b as
thick yellow syrup. (4.2 g, 91% for two steps). (DMSO, 400 MHz):
.delta. 8.17 (d, 1H), 7.96 (d, 1H), 3.83 (s, 3H), 3.17 (s, 3H),
1.36 (s, 9H).
Step 3:
[0362] To a solution of nitroarene 10b (4.2 g, 12.8 mmol) in
methanol (70 mL), was added palladium on carbon (Degussa, 10%, ca.
1.0 g). The mixture was purged and placed under an atmosphere of
hydrogen. It was stirred overnight, then purged with argon, and
filtered through a short pad of celite. The filtrate was
concentrated to afford aniline 10c as a colorless glass which was
used for the next step. (DMSO, 400 MHz): .delta. 7.59 (d, 1H), 6.85
(s, 2H), 6.57 (d, 1H), 3.77 (s, 3H), 3.01 (s, 3H), 1.24 (s,
9H).
Step 4:
[0363] To a solution of aniline 10c (12.8 mmol based on
theoretical) in DCM (50 mL), was added DIPEA (2.4 mL, 5.1 mmol),
followed by methyl 4-isocyanatobenzoate (2.50 g, 14.1 mmol). The
resulting mixture was stirred over the weekend during which time a
precipitate formed. The crude reaction mixture was filtered and
both the solid and filtrate contained the desired product plus
small amounts of impurities. The filtrate was then concentrated and
purified by silica gel chromatography (0-10% ethyl acetate/DCM) to
afford desired product 10d as a colorless oil (1.2g, 21%). MS
found: (M-H).sup.- 442.3.
Step 5:
[0364] Compound 10e was synthesized in a manner similar to Example
1, Step 5. MS found: (M-H).sup.- 428.3.
Step 6:
[0365] Compound 10f was synthesized in a manner similar to Example
1, Step 6. (DMSO, 400 MHz): .delta. 11.73 (s, 1H), 9.58 (d, 1H),
7.97 (d, 2H), 7.91 (d, 1H), 7.43 (d, 2H), 7.03 (m, 3H), 5.83 (d,
1H), 3.70 (s, 3H), 3.12 (s, 3H), 1.34 (s, 9H).
Step 7:
[0366] Compound 10g was synthesized in a manner similar to Example
1, Step 5 and was used immediately for the next step.
Step 8:
[0367] To a solution of carbamate 10g (135 mg, 0.22 mmol) in DCM (9
mL), was added trifluoroacetic acid (1 mL). The mixture was stirred
at rt for 2 hours, then concentrated and the crude product was
purified by HPLC to afford 48 as a white solid (85 mg, 76%). MS
found: (M-H).sup.- 501.2, 503.0.
Example 50
N-((5-chlorothiophen-2-yl)(1H-tetrazol-5-yl)methyl)-4-(7-fluoro-6-(methyla-
mino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide (Compound
49)
##STR00077##
[0369] Compound 49 was synthesized in a manner similar to Example
11 using intermediate 10e. MS found for Compound 49 as (M+H).sup.+:
527.2.
Example 51
N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)methyl)--
4-7-fluoro-6-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzam-
ide (Compound 50)
##STR00078##
[0371] Compound 50 was synthesized in a manner similar to Example
25 using intermediate 10e from Example 49. MS found for Compound 50
as (M+H).sup.+: 543.0.
Example 52
N-((1H-tetrazol-5-yl)(3-(trifluoromethyl)phenyl)methyl)-4-(6-fluoro-7-(met-
hylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide
(Compound 34)
##STR00079##
[0373] The titled compound was made by the procedure similar to
that described for Example 12. (DMSO, 400 MHz): .delta. 9.25 (d,
1H), 7.88 (d, 2H), 7.72 (s, 1H), 7.67 (d, 1H), 7.50 (d, 1H), 7.42
(d, 1H), 7.34 (s, 1H), 7.24 (d, 2H), 6.70 (s, 1H), 6.48 (d, 1H),
6.17 (d, 1H), 2.68 (d, 3H). MS found for Compound 34 as
(M+H).sup.+: 555.0.
Example 53
2-(5-chlorothiophen-2-yl)-2-(3-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2--
dihydroquinazolin-3(4H)-yl)phenyl)ureido)acetic acid (Compound
110)
##STR00080##
[0374] Step 1:
[0375] To a solution of aniline 11a (336 mg, 1.62 mmol) in DCM (10
mL), was added DIPEA (1.97 mL, 11.34 mmol). This solution was added
to a solution of phosgene (2 M in toluene, 2.43 mL, 4.86 mmol) in
an ice bath. Upon complete addition the ice bath was removed and
the reaction checked by HPLC which showed the formation of the
reactive intermediate. The crude reaction mixture was concentrated,
resuspended in DCM (10 mL), and the mixture was added to a solution
containing amino acid 11b (238 mg, 1.25 mmol) and DIPEA (0.43 mg,
2.5 mmol) in 15 mL of DMF. The reaction was stirred over the
weekend at room temperature, diluted with water and acidified with
1 M HCl to pH=2. The aqueous layer was extracted twice with ethyl
acetate. The combined organic layer was concentrated and purified
by silica gel chromatography (0-10% methanol/DCM) to afford 11c as
a light red oil containing a small amount of DMF. MS found:
(M-H).sup.- 424.0, 426.2.
Step 2:
[0376] Compound 11d was synthesized in a similar manner to Example
49, Step 8. MS found: (M+H).sup.+ 326.0, 328.0.
Step 3:
[0377] Compound 11e was synthesized in a similar manner to Example
29, Step 4 using DMF as solvent. MS found for
C.sub.22H.sub.17ClF.sub.2N.sub.4O.sub.6S as (M-H).sup.- 537.3,
539.0.
Step 4:
[0378] Compound 110 was prepared in a manner similar to Example 1,
Step 4. MS found: (M-H).sup.- 516.0, 518.0.
Examples 54-122
N-((5-chlorothiophen-2-yl)methyl)-4-(7-(4-fluorobenzylamino)-2,4-dioxo-1,2-
-dihydroquinazolin-3(4H)-yl)benzamide (Compound 1)
##STR00081##
[0379] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 535.0.
2-(4-(7-(4-fluorobenzylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)ben-
zamido)-2-(thiophen-2-yl)acetic acid (Compound 2)
##STR00082##
[0380] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 545.0.
2-(4-(7-(4-fluorobenzylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)ben-
zamido)-3-(thiophen-2-yl)propanoic acid (Compound 3)
##STR00083##
[0381] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 559.0.
2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)be-
nzamido)-2-(thiophen-2-yl)acetic acid (Compound 8)
##STR00084##
[0382] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 470.0.
2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)be-
nzamido)-3-(thiophen-2-yl)propanoic acid (Compound 9)
##STR00085##
[0383] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 483.0.
N-((1H-tetrazol-5-yl)methyl)-4-(7-(4-fluorobenzylamino)-2,4-dioxo-1,2-dihy-
droquinazolin-3(4H)-yl)benzamide (Compound 10)
##STR00086##
[0384] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 487.0.
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dih-
ydroquinazolin-3(4H)-yl)benzamido)acetic acid (Compound 13)
##STR00087##
[0385] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 503.0.
2-(4-chlorophenyl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroqui-
nazolin-3(4H)-yl)benzamido)acetic acid (Compound 14)
##STR00088##
[0386] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 497.0.
methyl
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-b-
enzo[e][1,3]oxazin-3(4H)-yl)benzamido)acetate (Compound 18)
##STR00089##
[0387] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 517.0.
2-(5-chlorothiophen-2-yl)-2-(4-(7-(methylamino)-2,4-dioxo-1,2-dihydroquina-
zolin-3(4H)-yl)benzamido)acetic acid (Compound 20)
##STR00090##
[0388] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 485.0.
ethyl
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1-
,2-dihydroquinazolin-3(4H)-yl)benzamido)acetate (Compound 26)
##STR00091##
[0389] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 531.0.
2-(5-chlorothiophen-2-yl)-2-(4-(2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)b-
enzamido)acetic acid (Compound 33)
##STR00092##
[0390] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 456.0.
2-morpholinoethyl
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-di-
hydroquinazolin-3(4H)-yl)benzamido)acetate (Compound 35)
##STR00093##
[0391] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.-617.0.
N-(1-(5-chlorothiophen-2-yl)-2-hydroxyethyl)-4-(6-fluoro-7-(methylamino)-2-
,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide (Compound 36)
##STR00094##
[0392] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 489.0.
2-(5-chlorothiophen-2-yl)-2-(4-(6-iodo-2,4-dioxo-1,2-dihydroquinazolin-3(4-
H)-yl)benzamido)acetic acid (Compound 37)
##STR00095##
[0393] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 581.7.
2-(5-chlorothiophen-2-yl)-2-(4-(6-isopropoxy-2,4-dioxo-1,2-dihydroquinazol-
in-3(4H)-yl)benzamido)acetic acid (Compound 38)
##STR00096##
[0394] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 515.0.
2-(5-chlorothiophen-2-yl)-2-(4-(2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)b-
enzamido)acetic acid (Compound 39)
##STR00097##
[0395] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 422.0.
N-(2-amino-1-(5-chlorothiophen-2-yl)-2-oxoethyl)-4-(6-fluoro-7-(methylamin-
o)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)benzamide (Compound
47)
##STR00098##
[0396] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 502.0.
(S)-2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benz-
o[e][1,3]oxazin-3(4H)-yl)benzamido)acetic acid (Compound 62)
##STR00099##
[0397] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 490.0.
methyl
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-b-
enzo[e][1,3]oxazin-3(4H)-yl)benzamido)acetate (Compound 66)
##STR00100##
[0398] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 504.0.
ethyl
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-be-
nzo[e][1,3]oxazin-3(4H)-yl)benzamido)acetate (Compound 67)
##STR00101##
[0399] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 518.0.
2-(5-chlorothiophen-2-yl)-2-(4-(8-methoxy-4-oxo-2H-benzo[e][1,3]oxazin-3(4-
H)-yl)benzamido)acetic acid (Compound 68)
##STR00102##
[0400] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 473.0.
2-(5-chlorothiophen-2-yl)-2-(4-(7-methyl-4-oxo-2H-benzo[e][1,3]oxazin-3(4H-
)-yl)benzamido)acetic acid (Compound 69)
##STR00103##
[0401] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 457.0.
N-(2-amino-1-(5-chlorothiophen-2-yl)-2-oxoethyl)-4-(6-fluoro-7-(methylamin-
o)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamide (Compound
70)
##STR00104##
[0402] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 489.0.
2-(5-chlorothiophen-2-yl)-2-(4-(6-ethyl-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-
-yl)benzamido)acetic acid (Compound 72)
##STR00105##
[0403] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 471.0.
N-((5-chlorothiophen-2-yl)(1-methyl-1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-
-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamide
(Compound 73)
##STR00106##
[0404] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 528.0.
N-((5-chlorothiophen-2-yl)(2-methyl-2H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-
-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamide
(Compound 74)
##STR00107##
[0405] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 528.0.
methyl
2-(5-chlorothiophen-2-yl)-2-(4-(6-methyl-4-oxo-2H-benzo[e][1,3]oxaz-
in-3(4H)-yl)benzamido)acetate (Compound 76)
##STR00108##
[0406] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 471.0.
2-(5-chlorothiophen-2-yl)-2-(4-(6-methyl-4-oxo-2H-benzo[e][1,3]oxazin-3(4H-
)-yl)benzamido)acetic acid (Compound 77)
##STR00109##
[0407] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 457.0.
2-(5-chlorothiophen-2-yl)-2-(4-(7-ethyl-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-
-yl)benzamido)acetic acid (Compound 78)
##STR00110##
[0408] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 471.0.
N-((3-chlorophenyl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(methylamino)-4-
-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamide (Compound 80)
##STR00111##
[0409] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 508.0.
N-((5-chlorothiophen-2-yl)(thiazol-2-yl)methyl)-4-(6-fluoro-7-(methylamino-
)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamide (Compound 82)
##STR00112##
[0410] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 529.0.
2-(5-chlorothiophen-2-yl)-2-(4-(4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benz-
amido)acetic acid (Compound 83)
##STR00113##
[0411] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 443.0.
2-(5-chlorothiophen-2-yl)-2-(4-(7-(4-fluorobenzylamino)-4-oxo-2H-benzo[e][-
1,3]oxazin-3(4H)-yl)benzamido)acetic acid (Compound 84)
##STR00114##
[0412] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 566.0.
N-((5-chlorothiophen-2-yl)(4-methylthiazol-2-yl)methyl)-4-(5-fluoro-7-(met-
hylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamide (Compound
85)
##STR00115##
[0413] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 544.0.
N-((5-chlorothiophen-2-yl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(methyla-
mino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamide (Compound
87)
##STR00116##
[0414] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 514.0.
(S)--N-((5-chlorothiophen-2-yl)(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)met-
hyl)-4-6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benz-
amide (Compound 88)
##STR00117##
[0415] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 530.0.
(R)--N-((5-chlorothiophen-2-yl)(1H-tetrazol-5-yl)methyl)-4-(6-fluoro-7-(me-
thylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)benzamide
(Compound 89)
##STR00118##
[0416] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 514.0.
2-(5-chlorothiophen-2-yl)-2-(4-(7-fluoro-6-(methylamino)-1-oxoisoquinolin--
2(1H)-yl)benzamido)acetic acid (Compound 90)
##STR00119##
[0417] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 486.0.
N-((5-chlorothiophen-2-yl)(1H-tetrazol-5-yl)methyl)-4-(7-fluoro-6-(methyla-
mino)-1-oxoisoquinolin-2(1H)-yl)benzamide (Compound 91)
##STR00120##
[0418] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 510.0.
2-(5-chlorothiophen-2-yl)-2-(4-(7-fluoro-6-methoxy-1-oxoisoquinolin-2(1H)--
yl)benzamido)acetic acid (Compound 92)
##STR00121##
[0419] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 487.0.
2-(5-chlorothiophen-2-yl)-2-(4-(6-ethyl-1-oxoisoquinolin-2(1H)-yl)benzamid-
o)acetic acid (Compound 93)
##STR00122##
[0420] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 467.0.
2-(4-(6-carbamoyl-1-oxoisoquinolin-2(1H)-yl)benzamido)-2-(5-chlorothiophen-
-2-yl)acetic acid (Compound 94)
##STR00123##
[0421] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 482.0.
2-(5-chlorothiophen-2-yl)-2-(4-(6-cyano-1-oxoisoquinolin-2(1H)-yl)benzamid-
o)acetic acid (Compound 95)
##STR00124##
[0422] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 430.0.
2-(5-chlorothiophen-2-yl)-2-(4-(1-oxoisoquinolin-2(1H)-yl)benzamido)acetic
acid (Compound 96)
##STR00125##
[0423] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 439.0.
2-(4-(1-oxoisoquinolin-2(1H)-yl)benzamido)-2-(thiophen-2-yl)acetic
acid (Compound 97)
##STR00126##
[0424] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 405.0.
2-(5-chlorothiophen-2-yl)-2-(4-(6-cyano-1-oxoisoquinolin-2(1H)-yl)benzamid-
o)acetic acid (Compound 98)
##STR00127##
[0425] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 464.0.
4-(4-carbamoyl-1-oxoisoquinolin-2(1H)-yl)benzamido)-2-(5-chlorothiophen-2--
yl)acetic acid (Compound 99)
##STR00128##
[0426] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 482.0.
2-(5-chlorothiophen-2-yl)-2-(4-(4-cyano-1-oxoisoquinolin-2(1H)-yl)benzamid-
o)acetic acid (Compound 100)
##STR00129##
[0427] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 464.0.
2-(5-chlorothiophen-2-yl)-2-(4-(6-methoxy-1-oxoisoquinolin-2(1H)-yl)benzam-
ido)acetic acid (Compound 101)
##STR00130##
[0428] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 469.0.
2-(4-(4-bromo-1-oxoisoquinolin-2(1H)-yl)benzamido)-2-(5-chlorothiophen-2-y-
l)acetic acid (Compound 102)
##STR00131##
[0429] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 518.0.
2-(5-chloro-N-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dihydroquinazolin-
-3(4H)-yl)benzyl)thiophene-2-carboxamido)acetic acid (Compound
103)
##STR00132##
[0430] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 517.0.
3-(5-chlorothiophen-2-yl)-3-(4-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dih-
ydroquinazolin-3(4H)-yl)benzylamino)propanoic acid (Compound
104)
##STR00133##
[0431] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 503.0.
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e]-
[1,3]oxazin-3(4H)-yl)benzylamino)acetic acid (Compound 105)
##STR00134##
[0432] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 476.0.
3-(5-chlorothiophen-2-yl)-3-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo[e]-
[1,3]oxazin-3(4H)-yl)benzylamino)propanoic acid (Compound 106)
##STR00135##
[0433] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 490.0.
1-((5-chlorothiophen-2-yl)methyl)-3-(4-(7-(4-fluorobenzylamino)-2,4-dioxo--
1,2-dihydroquinazolin-3(4H)-yl)phenyl)urea (Compound 107)
##STR00136##
[0434] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 551.0.
2-(3-(4-(7-(4-fluorobenzylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)-
phenyl)ureido)-2-(thiophen-2-yl)acetic acid (Compound 108)
##STR00137##
[0435] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 559.2.
2-(3-(4-(7-(4-fluorobenzylamino)-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)-
phenyl)ureido)-3-(thiophen-2-yl)propanoic acid (Compound 109)
##STR00138##
[0436] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 574.0.
2-(5-chlorothiophen-2-yl)-2-(5-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dih-
ydroquinazolin-3(4H)-yl)thiophene-2-carboxamido)acetic acid
(Compound 112)
##STR00139##
[0437] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 509.0.
2-(5-chlorothiophen-2-yl)-2-(5-(6-fluoro-7-(methylamino)-2,4-dioxo-1,2-dih-
ydroquinazolin-3(4H)-yl)thiophene-3-carboxamido)acetic acid
(Compound 113)
##STR00140##
[0438] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 509.0.
2-(5-chlorothiophen-2-yl)-2-(2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo-
[e][1,3]oxazin-3(4H)-yl)phenyl)acetamido)acetic acid (Compound
114)
##STR00141##
[0439] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 504.0.
2-(5-chlorothiophen-2-yl)-2-(4-(7-fluoro-6-(methylamino)-1-oxoisoquinolin--
2(1H)-yl)benzylamino)acetic acid (Compound 115)
##STR00142##
[0440] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 472.0.
2-(5-chlorothiophen-2-yl)-2-(4-(6-fluoro-7-(methylamino)-4-oxoquinazolin-3-
(4H)-yl)benzamido)acetic acid (Compound 116)
##STR00143##
[0441] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 487.0.
2-(5-chlorothiophen-2-yl)-2-(4-(7-fluoro-6-(methylamino)-1-oxophthalazin-2-
(1H)-yl)benzamido)acetic acid (Compound 117)
##STR00144##
[0442] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 487.0.
methyl
2-(5-chlorothiophen-2-yl)-2-(2-(4-(6-fluoro-7-(methylamino)-4-oxo-2-
H-benzo[e][1,3]oxazin-3(4H)-yl)phenyl)acetamido)acetate (Compound
118)
##STR00145##
[0443] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 518.0.
2-(5-chlorothiophen-2-yl)-2-(4-(6-ethyl-1-oxo-3,4-dihydroisoquinolin-2(1H)-
-yl)benzamido)acetic acid (Compound 119)
##STR00146##
[0444] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 469.0.
2-(5-chlorothiophen-2-yl)-2-(2-(4-(6-fluoro-7-(methylamino)-4-oxo-2H-benzo-
[e][1,3]oxazin-3(4H)-yl)phenyl)acetamido)-N,N-dimethylacetamide
(Compound 120)
##STR00147##
[0445] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 558.0.
N-(1-(5-chlorothiophen-2-yl)-2-oxo-2-(pyrrolidin-1-yl)ethyl)-2-(4-(6-fluor-
o-7-(methylamino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)phenyl)acetamide
(Compound 121); and
##STR00148##
[0446] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 558.0.
N-(2-amino-1-(5-chlorothiophen-2-yl)-2-oxoethyl)-2-(4-(6-fluoro-7-(methyla-
mino)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)phenyl)acetamide
(Compound 122)
##STR00149##
[0447] was prepared in a manner similar to Examples above. MS
found: (M-H).sup.- 503.0.
Example 123
Pharmacological Assays
[0448] The pharmacological activity of each of the compounds
according to the invention is determined by the following in vitro
assays:
I. Inhibition of ADP-Mediated Platelet Aggregation In Vitro
[0449] 1.
[0450] The effect of testing the compound according to the
invention on ADP-induced human platelet aggregation was assessed in
a 96-well microtiter assay (see generally the procedures in
Jantzen, H. M. et al. (1999) Thromb. Hemost. 81:111-117) or
standard cuvette light transmittance aggregometry using either
human platelet-rich plasma (PRP) or human washed platelets.
[0451] For preparation of human platelet-rich plasma for
aggregation assays, human venous blood was collected from healthy,
drug-free volunteers into 0.38% sodium citrate (0.013 M, pH 7.0
final). Platelet-rich plasma (PRP) is prepared by centrifugation of
whole blood at 160.times.g for 20 minutes at room temperature. The
PRP layer is removed, transferred to a new tube, and the platelet
count is adjusted, if necessary, to achieve a platelet
concentration of .about.3.times.10.sup.8 platelets/ml using
platelet-poor plasma (PPP). PPP is prepared by centrifugation of
the remaining blood sample (after removal of PRP) for 20 minutes at
800.times.g. This preparation of PRP can subsequently be used for
aggregation assays in either a 96-well plate or standard cuvette
aggregometry.
[0452] For preparation of washed platelets, human venous blood is
collected from healthy, drug-free volunteers into ACD (85 mM sodium
citrate, 111 mM glucose, 71.4 mM citric acid) containing PGI.sub.2
(1.25 ml ACD containing 0.2 .mu.M PGI2 final; PGI.sub.2 was from
Sigma, St. Louis, Mo.). Platelet-rich plasma (PRP) is prepared by
centrifugation at 160.times.g for 20 minutes at room temperature.
Washed platelets are prepared by centrifuging PRP for 10 minutes at
730 g and resuspending the platelet pellet in CGS (13 mM sodium
citrate, 30 mM glucose, 120 mM NaCl; 2 ml CGS/10 ml original blood
volume) containing 1 U/ml apyrase (grade V, Sigma, St. Louis, Mo.).
After incubation at 37.degree. C. for 15 minutes, the platelets are
collected by centrifugation at 730 g for 10 minutes and resuspended
at a concentration of 3.times.10.sup.8 platelets/ml in
Hepes-Tyrode's buffer (10 mM Hepes, 138 mM NaCl, 5.5 mM glucose,
2.9 mM KCl, 12 mM NaHCO.sub.3, pH 7.4) containing 0.1% bovine serum
albumin, 1 mM CaCl.sub.2 and 1 mM MgCl.sub.2. This platelet
suspension is kept >45 minutes at 37.degree. C. before use in
aggregation assays.
2.
[0453] For cuvette light transmittance aggregation assays, serial
dilutions (1:3) of test compounds were prepared in 100% DMSO in a
96 well V-bottom plate (final DMSO concentration in the cuvette was
0.6%). The test compound (3 .mu.l of serial dilutions in DMSO) was
preincubated with PRP for 30-45 seconds prior to initiation of
aggregation reactions, which were performed in a ChronoLog
aggregometer by addition of agonist (5 or 10 .mu.M ADP) to 490
.mu.L of PRP at 37.degree. C. In some cases, light transmittance
aggregometry was performed using 490 .mu.L of washed platelets
(prepared as described above) at 37.degree. C., and aggregation was
initiated by addition of 5 .mu.M ADP and 0.5 mg/ml human fibrinogen
(American Diagnostics, Inc., Greenwich, Conn.). The aggregation
reaction is recorded for .about.5 min, and maximum extent of
aggregation is determined by the difference in extent of
aggregation at baseline, compared to the maximum aggregation that
occurs during the five minute period of the assay. Inhibition of
aggregation was calculated as the maximum aggregation observed in
the presence of inhibitor, compared to that in the absence of
inhibitor. IC.sub.50s were derived by non-linear regression
analysis using the Prism software (GraphPad, San Diego,
Calif.).
3.
[0454] Inhibition of ADP-dependent aggregation was also determined
in 96-well flat-bottom microtiter plates using a microtiter plate
shaker and plate reader similar to the procedure described by
Frantantoni et al., Am. J. Clin. Pathol. 94, 613 (1990). All steps
are performed at room temperature. For 96-well plate aggregation
using platelet-rich plasma (PRP), the total reaction volume of 0.2
ml/well includes 180 .mu.l of PRP (.about.3.times.108 platelets/ml,
see above), 6 .mu.l of either serial dilution of test compounds in
20% DMSO or buffer (for control wells), and 10 .mu.l of 20.times.
ADP agonist solution (100 .mu.M). The OD of the samples is then
determined at 450 nm using a microtiter plate reader (Softmax,
Molecular Devices, Menlo Park, Calif.) resulting in the 0 minute
reading. The plates are then agitated for 5 min on a microtiter
plate shaker and the 5 minute reading is obtained in the plate
reader. Aggregation is calculated from the decrease of OD at 450 nm
at t=5 minutes compared to t=0 minutes and is expressed as % of the
decrease in the ADP control samples after correcting for changes in
the unaggregated control samples. IC.sub.50s were derived by
non-linear regression analysis.
[0455] For 96-well plate aggregation using washed platelets, the
total reaction volume of 0.2 ml/well includes in Hepes-Tyrodes
buffer/0.1% BSA: 4.5.times.10.sup.7 apyrase-washed platelets, 0.5
mg/ml human fibrinogen (American Diagnostica, Inc., Greenwich,
Conn.), serial dilutions of test compounds (buffer for control
wells) in 0.6% DMSO. After .about.5 minutes preincubation at room
temperature, ADP is added to a final concentration of 2 .mu.M which
induces submaximal aggregation. Buffer is added instead of ADP to
one set of control wells (ADP-control). The OD of the samples is
then determined at 450 nm using a microtiter plate reader (Softmax,
Molecular Devices, Menlo Park, Calif.) resulting in the 0 minute
reading. The plates are then agitated for 5 min on a microtiter
plate shaker and the 5 minute reading is obtained in the plate
reader. Aggregation is calculated from the decrease of OD at 450 nm
at t=5 minutes compared to t=0 minutes and is expressed as % of the
decrease in the ADP control samples after correcting for changes in
the unaggregated control samples. IC.sub.50s were derived by
non-linear regression analysis.
II. Inhibition of [3H]2-MeS-ADP Binding to Platelets
1. The Ability of Candidate Molecules to Inhibit the Binding of
[3H12-MeS-ADP to the P2Y12 Receptor on Platelets was Determined
Using a Radioligand Binding Assay.
[0456] Utilizing this assay the potency of inhibition of such
compounds with respect to [.sup.3H]2-MeS-ADP binding to whole
platelets is determined. Under the conditions described in
[0457] II (3) below, the binding of [.sup.3H]2-MeS-ADP is solely
due to the interaction of this ligand with the P2Y.sub.12 receptor,
in that all the specific binding measured in this assay is
competable with a P2Y.sub.12 antagonist (i.e., the specific binding
is reduced to background levels by competition with an excess of
P2Y.sub.12 antagonist, with no competition of binding when a
P2Y.sub.1 antagonist is pre-incubated with the platelet
preparation). [.sup.3H]2-MeS-ADP binding experiments are routinely
performed with outdated human platelets collected by standard
procedures at hospital blood banks Apyrase-washed outdated
platelets are prepared as follows (all steps at room temperature,
if not indicated otherwise):
[0458] Outdated platelet suspensions are diluted with 1 volume of
CGS and platelets pelleted by centrifugation at 1900.times.g for 45
minutes. Platelet pellets are resuspended at 3-6.times.10.sup.9
platelets/ml in CGS containing 1 U/ml apyrase (grade V, Sigma, St.
Louis, Mo.) and incubated for 15 minutes at 37.degree. C. After
centrifugation at 730.times.g for 20 minutes, pellets are
resuspended in Hepes-Tyrode's buffer containing 0.1% BSA (Sigma,
St. Louis, Mo.) at a concentration of 6.66.times.10.sup.8
platelets/ml. Binding experiments are performed after >45
minutes resting of the platelets.
2.
[0459] Alternatively, binding experiments are performed with fresh
human platelets prepared as described in section I (Inhibition of
ADP-Mediated Platelet Aggregation in vitro), except that platelets
are resuspended in Hepes-Tyrode's buffer containing 0.1% BSA
(Sigma, St. Louis, Mo.) at a concentration of 6.66.times.10.sup.8
platelets/mil. Very similar results are obtained with fresh and
outdated platelets.
3.
[0460] A platelet ADP receptor binding assay (ARB) using the
tritiated potent agonist ligand [.sup.3H]2-MeS-ADP (Jantzen, H. M.
et al. (1999) Thromb. Hemost. 81:111-117) has been adapted to the
96-well microtiter format. In an assay volume of 0.2 ml
Hepes-Tyrode's buffer with 0.1% BSA and 0.6% DMSO, 1.times.10.sup.8
apyrase-washed platelets are preincubated in 96-well flat bottom
microtiter plates for 5 minutes with serial dilutions of test
compounds before addition of 1 nM [.sup.3H]2-MeS-ADP
([.sup.3H]2-methylthioadenosine-5'-diphosphate, ammonium salt;
specific activity 20-50 Ci/mmole, obtained by custom synthesis from
Amersham Life Science, Inc., Arlington Heights, Ill., or NEN Life
Science Products, Boston, Mass.). Total binding is determined in
the absence of test compounds. Samples for nonspecific binding may
contain 10 .mu.M unlabelled 2-MeS-ADP (RBI, Natick, Mass.). After
incubation for 15 minutes at room temperature, unbound radioligand
is separated by rapid filtration and two washes with cold
(4-8.degree. C.) Binding Wash Buffer (10 mM Hepes pH 7.4, 138 mM
NaCl) using a 96-well cell harvester (Minidisc 96, Skatron
Instruments, Sterling, Va.) and 8.times.12 GF/C glassfiber
filtermats (Printed Filtermat A, for 1450 Microbeta, Wallac Inc.,
Gaithersburg, Md.). The platelet-bound radioactivity on the
filtermats is determined in a scintillation counter (Microbeta
1450, Wallac Inc., Gaithersburg, Md.). Specific binding is
determined by subtraction of non-specific binding from total
binding, and specific binding in the presence of test compounds is
expressed as % of specific binding in the absence of test compound
dilutions. IC.sub.50s were derived by non-linear regression
analysis.
[0461] In the table below, activity in the PRP assay is provided as
follows: +++, IC.sub.50<10 .mu.M; ++, 10
.mu.M<IC.sub.50<30 .mu.M; and +, IC.sub.50>30 .mu.M.
Activity in the ARB assay is provided as follows: +++,
IC.sub.50<0.05 .mu.M; ++, 0.05 .mu.M<IC.sub.50<0.5 .mu.M;
and +, IC.sub.50>0.5 .mu.M.
TABLE-US-00001 TABLE 1 Activity of the compounds in ARB and PRP
assays Compound No. ARB (.mu.M) PRP (.mu.M) 1 ++ + 2 ++ + 3 ++ 4 ++
5 +++ ++ 6 ++ + 7 +++ +++ 8 + 9 + 10 + 11 ++ + 12 + 13 ++ +++ 14 +
15 ++ 16 ++ +++ 18 + 19 ++ +++ 20 ++ 21 +++ +++ 22 + 23 + 24 + 25
++ 26 + 27 ++ + 28 + 29 + 30 + 31 + 32 ++ +++ 33 + 34 + 35 ++ ++ 36
++ 37 ++ 38 + 39 + 40 + 41 ++ ++ 42 + 43 ++ 44 ++ ++ 45 +++ +++ 46
++ ++ 47 ++ +++ 48 ++ 49 + 50 ++ + 51 + 52 ++ 53 ++ + 54 +++ ++ 55
++ + 56 +++ ++ 57 ++ 58 ++ 59 + 60 ++ 61 +++ ++ 62 +++ + 63 + 64 ++
65 ++ 66 ++ 67 ++ 68 + 69 + 70 ++ ++ 71 +++ ++ 72 + 73 ++ 74 ++ +
75 +++ ++ 76 + 77 + 78 ++ 79 ++ +++ 80 ++ + 81 ++ + 82 ++ + 83 + 84
+ 85 ++ 86 + 87 +++ 88 +++ 89 +++ 90 +++ ++ 91 +++ ++ 92 + 93 + 94
+ 95 + 96 + 97 + 98 + 99 + 100 + 101 + 102 + 103 + 104 ++ 105 ++ +
106 ++ + 107 +++ 108 ++ 109 ++ 110 + 111 + 112 ++ 113 + 114 + 115
++ + 116 ++ 117 ++ 118 ++ 119 + 120 + 121 + 122 ++
[0462] It should be understood that the foregoing discussion,
embodiments and examples merely present a detailed description of
certain preferred embodiments. It will be apparent to those of
ordinary skill in the art that various modifications and
equivalents can be made without departing from the spirit and scope
of the invention. All the patents, journal articles and other
documents discussed or cited above are herein incorporated by
reference.
* * * * *