Note: Descriptions are shown in the official language in which they were submitted.
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
TREATMENT OF CARDIOVASCULAR DISEASE AND DYSLIPIDEMIA
USING SECRETORY PHOSPHOLIPASE A2 (SPLA2) INHIBITORS AND
SPLA2 INHIBITOR COMBINATION THERAPIES
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] The subject application claims the benefit of U.S. Application No.
11/874,869, filed October 18, 2007, U.S. Provisional Patent Application No.
60/969,591, filed August 31, 2007, and U.S. Provisional Patent Application
No. 60/915,910, filed May 3, 2007. The disclosures of each of these
applications are incorporated herein by reference in their entirety, including
drawings.
BACKGROUND
[0002] In 2004, it was estimated that over 75 million Americans had one or
more forms of cardiovascular disease (CVD). Coronary heart disease (CHD)
and coronary artery disease (CAD) are the most common types of CVD. CHD
and CAD occur when coronary arteries that supply blood to the heart become
hardened and narrowed due to atherosclerosis. A variety of therapeutic
options are currently employed in the treatment of CVD and conditions
associated with CVD. Many of these therapeutic options function by lowering
cholesterol levels, particularly LDL levels. Among the most popular and
effective of these therapeutic options are statins, a class of compounds that
inhibit cholesterol biosynthesis and prevent the build-up of arterial plaque.
Statin administration has been shown to lower LDL and triglyceride levels and
to substantially reduce coronary events and death frorn CVD. However, statin
therapy alone is insufficient to completely treat CVD. Therefore, there is a
need in the art for more effective methods of treating CVD and conditions
associated with CVD.
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
SUMMARY
[0003] In certain embodiments, methods are provided for treating
dyslipidemia in a subject in need thereof by administering a therapeutically
effective amount of one or more sPLA2 inhibitors. In certain embodiments, the
one or more sPLA2 inhibitors are administered in a composition that further
comprises one or more pharmaceutically acceptable carriers. In certain
embodiments, the one or more sPLA2 inhibitors comprise A-001 or a
pharmaceutically acceptable prodrug, salt, polymorph, co-crystal, or solvate
of
A-001. In certain of these embodiments, the one or more sPLA2 inhibitors
comprise A-002, a prodrug of A-001. In certain embodiments, administration
of one or more sPLA2 inhibitors results in a decrease in cholesterol and/or
triglyceride levels. In certain of these embodiments, administration of one or
more sPLA2 inhibitors results in a decrease in total cholesterol, non-HDL
cholesterol, LDL, LDL particle, small LDL particle, oxidized LDL, and/or ApoB
levels. In certain embodiments, administration of one or more sPLA2
inhibitors results in an increase in HDL levels and/or LDL particle size. In
certain embodiments, administration of one or more sPLA2 inhibitors results in
a decrease in levels of one or more inflammatory markers. In certain of these
embodiments, the inflammatory markers may include, but are not limited to,
sPLA2, CRP, and/or IL-6. In certain embodiments, administration of one or
more sPLA2 inhibitors results in an improvement in HDL/LDL ratio. In certain
embodiments, the one or more sPLA2 inhibitors may be administered to the
subject twice or more per day, and in certain of these embodiments the one or
more sPLA2 inhibitors are administered twice per day. In other embodiments,
the one or more sPLA2 inhibitors may be administered to the subject on a
2
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
once a day basis. In certain embodiments, the one or more sPLA2 inhibitors
may be administered at a dosage of about 50 to about 500 mg.
[0004] In certain embodiments, methods are provided for decreasing
cholesterol levels in a subject in need thereof by administering a
therapeutically effective amount of one or more sPLA2 inhibitors. In certain
embodiments, the one or more sPLA2 inhibitors are administered in a
composition that further comprises one or more pharmaceutically acceptable
carriers. In certain embodiments, the one or more sPLA2 inhibitors comprise
A-001 or a pharmaceutically acceptable prodrug, salt, polymorph, co-crystal,
or solvate of A-001. In certain of these embodiments, the one or more sPLA2
inhibitors comprise A-002, a prodrug of A-001. In certain embodiments,
administration of one or more sPLA2 inhibitors results in a decrease in total
cholesterol, non-HDL cholesterol, LDL, LDL particle, small LDL particle,
oxidized LDL, and/or ApoB levels. In certain embodiments, administration of
one or more sPLA2 inhibitors results in an improvement in HDL/LDL ratio. In
certain embodiments, the one or more sPLA2 inhibitors may be administered
to the subject twice or more per day, and in certain of these embodiments the
one or more sPLA2 inhibitors are administered twice per day. In other
embodiments, the one or more sPLA2 inhibitors may be administered to the
subject on a once a day basis. In certain embodiments, the one or more
sPLA2 inhibitors may be administered at a dosage of about 50 to about 500
mg.
[0005] In certain embodiments, methods are provided for decreasing
triglyceride levels in a subject in need thereof by administering a
therapeutically effective amount of one or more sPLA2 inhibitors. In certain
3
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
embodiments, the one or more sPLA2 inhibitors are administered in a
composition that further comprises one or more pharmaceutically acceptable
carriers. In certain of these embodiments, the one or more sPLA2 inhibitors
comprise A-001 or a pharmaceutically acceptable prodrug, salt, pofyrnorph,
co-crystal, or solvate of A-001. In certain of these embodiments, the one or
more sPLA2 inhibitors comprise A-002, a prodrug of A-001. In certain
embodiments, the one or more sPLA2 inhibitors may be administered to the
subject twice or more per day, and in certain of these embodiments the one or
more sPLA2 inhibitors are administered twice per day. In other embodiments,
the one or niore sPLA2 inhibitors may be administered to the subject on a
once a day basis. In certain embodiments, the one or more sPLA2 inhibitors
may be administered at a dosage of about 50 to about 500 mg.
[0006] In certain embodiments, methods are provided for increasing HDL
levels in a subject in need thereof by administering a therapeutically
effective
amount of one or more sPLA2 inhibitors. In certain embodiments, the one or
more sPLA2 inhibitors are administered in a composition that further
comprises one or more pharmaceutically acceptable carriers. In certain
embodiments, the one or more sPLA2 inhibitors comprise A-001 or a
pharmaceutically acceptable prodrug, salt, polymorph, co-crystal, or solvate
of
A-001. In certain of these embodiments, the one or more sPLA2 inhibitors
comprise A-002, a prodrug of A-001. In certain embodiments, administration
of one or more sPLA2 inhibitors results in an improvement in HDL/LDL ratio.
In certain embodiments, improvement in HDL/LDL ratio is further
accomplished by a reduction in LDL levels. In certain ernbodiments, the one
or more sPLA2 inhibitors may be administered to the subject twice or more per
4
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
day, and in certain of these embodiments the one or more sPLA2 inhibitors
are administered twice per day. In other embodiments, the one or more
sPLA2 inhibitors may be administered to the subject on a once a day basis. In
certain embodiments, the one or more sPLA2 inhibitors may be administered
at a dosage of about 50 to about 500 mg.
[0007] In certain embodiments, methods are provided for treating CVD or a
condition associated with CVD in a subject in need thereof by administering a
therapeutically effective amount of one or more sPLA2 inhibitors. In certain
embodiments, the one or more sPLA2 inhibitors are administered in a
composition that further comprises one or more pharmaceutically acceptable
carriers. In certain embodiments, the one or more sPLA2 inhibitors comprise
A-001 or a pharmaceutically acceptable prodrug, salt, polymorph, co-crystal,
or solvate of A-001. In certain of these embodiments, the one or more sPLA2
inhibitors comprise A-002, a prodrug of A-001. In certain embodiments, the
one or more sPLA2 inhibitors may be administered to the subject twice or
more per day, and in certain of these embodiments the one or more sPLA2
inhibitors are administered twice per day. In other embodiments, the one or
more sPLA2 inhibitors may be administered to the subject on a once a day
basis. In certain embodiments, the one or more sPLA2 inhibitors may be
administered at a dosage of about 50 to about 500 mg. In certain
embodiments, administration of one or more sPLA2 inhibitors results in a
decrease in levels of one or more inflammatory markers. In certain of these
embodiments, the inflammatory markers may include, but are not lirnited to,
sPLA2, CRP, and/or IL-6. In certain embodiments, CVD or a condition
associated with CVD includes, but is not limited to, atherosclerosis,
metabolic
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
syndrome, coronary artery disease, coronary heart disease, cerebrovascular
disease, peripheral vascular disease, and/or conditions associated with
coronary artery disease, coronary heart disease, cerebrovascular disease, or
peripheral vascular disease.
[0008] In certain embodiments, methods are provided for treating metabolic
syndrome in a subject in need thereof by administering a therapeutically
effective amount of one or more sPLA2 inhibitors. In certain embodiments, the
one or more sPLA2 inhibitors are administered in a composition that further
comprises one or more pharmaceutically acceptable carriers. In certain
embodiments, the one or more sPLA2 inhibitors comprise A-001 or a
pharmaceutically acceptable prodrug, salt, polymorph, co-crystal, or solvate
of
A-001. In certain of these embodiments, the one or more sPLA2 inhibitors
comprise A-002, a prodrug of A-001. In certain embodiments, the one or
more sPLA2 inhibitors may be administered to the subject twice or more per
day, and in certain of these embodiments the one or more sPLA2 inhibitors
are administered twice per day. In other embodiments, the one or more
sPLA2 inhibitors may be administered to the subject on a once a day basis. In
certain embodiments, the one or more sPLA2 inhibitors may be administered
at a dosage of about 50 to about 500 mg. In certain embodiments,
administration of one or more sPLA2 inhibitors results in a decrease in levels
of one or more inflammatory markers. In certain of these embodiments, the
inflammatory markers may include, but are not limited to, sPLA2, CRP, and/or
IL-6.
[0009] In certain embodinients, compositions are provided comprising one or
more sPLA2 inhibitors and one or more compounds used in the treatment of
6
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
CVD. In certain embodiments, the composition further comprises one or more
pharmaceutically acceptable carriers. In certain embodiments, the one or
more sPLA2 inhibitors comprise A-001 or a pharmaceutically acceptable
prodrug, salt, polymorph, co-crystal, or solvate of A-001. In certain of these
embodiments, the one or more sPLA2 inhibitors comprise A-002, a prodrug of
A-001. In certain embodiments, the one or more compounds used in the
treatment of CVD comprise one or more statins or statin combination drugs.
In certain of these embodiments, the one or more statins are selected from
the group consisting of atorvastatin, simvastatin, rosuvastatin, lovastatin,
pravastatin, cerivastatin, fluvastatin, mevastatin, pitavastatin, and various
salts, solvates, stereoisomers, and prodrug derivatives thereof. In certain of
these embodiments, the statin combination drugs are selected from the group
consisting of atorvastatin plus ezetimibe, atorvastatin plus amlodipine,
atorvastatin plus CP-529414, atorvastatin plus APA-01, simvastatin plus
ezetimibe, simvastatin plus extended release niacin, simvastatin plus MK-
0524A, lovastatin plus extended release niacin, rosuvastatin plus fenofibrate,
pravastatin plus fenofibrate, and statin plus TAK-457. In certain
embodiments, the one or more compounds used in the treatment of CVD
comprise one or more non-statin compounds selected from the group
consisting of bile acid sequestrants, fibrates, niacin or niacin derivatives,
cholesterol absorption inhibitors, cholesteryl ester transfer protein (CETP)
inhibitors, microsomal triglyceride transfer protein (MTP) inhibitors,
squalene
synthase inhibitors, ACE inhibitors, angiotensin II receptor antagonists, beta-
adrenergic blockers, calcium channel blockers, and antithrombotics.
7
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
[0010] In certain embodiments, methods are provided for treating
dyslipidemia in a subject in need thereof by administering a therapeutically
effective amount of one or more sPLA2 inhibitors and a therapeutically
effective amount of one or more compounds used in the treatment of CVD. In
certain embodiments, the one or more sPLA2 inhibitors comprise A-001 or a
pharmaceutically acceptable prodrug, salt, polymorph, co-crystal, or solvate
of
A-001. In certain of these embodiments, the one or more sPLA2 inhibitors
comprise A-002, a prodrug of A-001. In certain embodiments, the one or
more compounds used in the treatment of CVD comprise one or more statins
or statin combination drugs. In certain of these embodiments, the one or
more statins are selected from the group consisting of atorvastatin,
simvastatin, rosuvastatin, lovastatin, pravastatin, cerivastatin, fluvastatin,
mevastatin, pitavastatin, and various salts, solvates, stereoisomers, and
prodrug derivatives thereof. In certain of these embodiments, the statin
combination drugs are selected from the group consisting of atorvastatin plus
ezetimibe, atorvastatin plus amlodipine, atorvastatin plus CP-529414,
atorvastatin plus APA-01, simvastatin plus ezetimibe, simvastatin plus
extended release niacin, simvastatin plus MK-0524A, lovastatin plus extended
release niacin, rosuvastatin plus fenofibrate, pravastatin plus fenofibrate,
and
statin plus TAK-457. In certain embodiments, the one or more compounds
used in the treatment of CVD comprise one or more non-statin compounds
selected from the group consisting of bile acid sequestrants, fibrates, niacin
or
niacin derivatives, cholesterol absorption inhibitors, cholesteryl ester
transfer
protein (CETP) inhibitors, microsomal triglyceride transfer protein (MTP)
inhibitors, squalene synthase inhibitors, ACE inhibitors, angiotensin II
receptor
8
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
antagonists, beta-adrenergic blockers, calcium channel blockers, and
antithrombotics. In certain embodiments, administration of one or more sPLA2
inhibitors and one or more compounds used in the treatment of CVD results in
a decrease in cholesterol levels and/or triglyceride levels. In certain of
these
embodiments, administration of one or more sPLA2 inhibitors and one or more
compounds used in the treatment of CVD results in a decrease in total
cholesterol, non-HDL cholesterol, LDL, LDL particle, small LDL particle,
oxidized LDL, and/or ApoB levels. In certain embodiments, administration of
one or more sPLA2 inhibitors and one or more compounds used in the
treatment of CVD results in an increase in HDL levels and/or LDL particle
size. In certain embodiments, administration of one or more sPLA2 inhibitors
and one or more compounds used in the treatment of CVD results in an
improvement in HDL/LDL ratio. In certain embodiments, the one or more
sPLA2 inhibitors and the one or more compounds used in the treatment of
CVD may be administered simultaneously. In certain of these embodiments,
the one or more sPLA2 inhibitors and the one or more compounds used in the
treatment of CVD may be administered in a single formulation, while in other
embodiments the compounds may be administered simultaneously in two or
more formulations. In each of these embodiments, the formulation(s) may
further comprise one or more pharmaceutically acceptable carriers. In other
embodiments, the one or more sPLA2 inhibitors and the one or more
compounds used in the treatment of CVD may be administered sequentially.
In certain embodiments, the one or more sPLA2 inhibitors may be
administered to the subject twice or more per day, and in certain of these
embodiments the one or more sPLA2 inhibitors are administered twice per
9
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
day. In other embodiments, the one or more sPLA2 inhibitors may be
administered to the subject on a once a day basis. In certain embodiments,
the one or more sPLA2 inhibitors may be administered at a dosage of about
50 to about 500 mg. In certain embodiments, administration of one or more
sPLA2 inhibitors and one or more compounds used in the treatment of CVD
results in a decrease in levels of one or more inflammatory markers. In
certain of these embodiments, the inflammatory markers may include, but are
not limited to, sPLA2, CRP, and/or IL-6.
[0011] In certain embodiments, methods are provided for decreasing
cholesterol levels in a subject in need thereof by administering a
therapeutically effective amount of one or more sPLA2 inhibitors and a
therapeutically effective amount of one or more compounds used in the
treatment of CVD. In certain embodiments, the one or more sPLA2 inhibitors
comprise A-001 or a pharmaceutically acceptable prodrug, salt, polymorph,
co-crystal, or solvate of A-001. In certain of these embodiments, the one or
more sPLA2 inhibitors comprise A-002, a prodrug of A-001. In certain
embodiments, the one or more compounds used in the treatment of CVD
comprise one or more statins or statin combination drugs. In certain of these
embodiments, the one or more statins are selected from the group consisting
of atorvastatin, simvastatin, rosuvastatin, lovastatin, pravastatin,
cerivastatin,
fluvastatin, mevastatin, pitavastatin, and various salts, solvates,
stereoisomers, and prodrug derivatives thereof. In certain of these
embodiments, the statin combination drugs are selected from the group
consisting of atorvastatin plus ezetimibe, atorvastatin plus amlodipine,
atorvastatin plus CP-529414, atorvastatin plus APA-01, simvastatin plus
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
ezetimibe, simvastatin plus extended release niacin, simvastatin plus MK-
0524A, lovastatin plus extended release niacin, rosuvastatin plus fenofibrate,
pravastatin plus fenofibrate, and statin plus TAK-457. In certain
embodiments, the one or more compounds used in the treatment of CVD
comprise one or more non-statin compounds selected from the group
consisting of bile acid sequestrants, fibrates, niacin or niacin derivatives,
cholesterol absorption inhibitors, cholesteryl ester transfer protein (CETP)
inhibitors, microsomal triglyceride transfer protein (MTP) inhibitors,
squalene
synthase inhibitors, ACE inhibitors, angiotensin II receptor antagonists, beta-
adrenergic blockers, calcium channel blockers, and antithrombotics. In
certain embodiments, administration of one or more sPLA2 inhibitors and one
or more compounds used in the treatment of CVD results in a decrease in
total cholesterol, non-HDL cholesterol, LDL, LDL particle, small LDL particle
levels, oxidized LDL, and/or ApoB levels_ In certain embodiments,
administration of one or more sPLA2 inhibitors and one or more compounds
used in the treatment of CVD results in an increase in LDL particle size. In
certain embodiments, adrninistration of one or more sPLA2 inhibitors and one
or more compounds used in the treatment of CVD results in an improvement
in HDL/LDL ratio. In certain embodiments, the one or more sPLA2 inhibitors
and the one or more compounds used in the treatment of CVD may be
administered simultaneously. In certain of these embodiments, the one or
more sPLA2 inhibitors and the one or more compounds used in the treatment
of CVD may be administered in a single formulation, while in other
embodiments the compounds may be administered simultaneously in two or
more fon-nulations. In each of these embodiments, the formulation(s) may
11
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
further comprise one or more pharmaceutically acceptable carriers. In other
embodiments, the one or more sPLA2 inhibitors and the one or more
compounds used in the treatment of CVD may be administered sequentially.
In certain embodiments, the one or more sPLA2 inhibitors may be
administered to the subject twice or more per day, and in certain of these
embodiments the one or more sPLA2 inhibitors are administered twice per
day. In other embodiments, the one or more sPLA2 inhibitors may be
administered to the subject on a once a day basis. In certain embodiments,
the one or more sPLA2 inhibitors may be administered at a dosage of about
50 to about 500 mg.
[0012] In certain embodiments, methods are provided for decreasing
triglyceride levels in a subject in need thereof by administering a
therapeutically effective amount of one or more sPLA2 inhibitors and a
therapeutically effective amount of one or more compounds used in the
treatment of CVD. In certain embodiments, the one or more sPLA2 inhibitors
comprise A-001 or a pharmaceutically acceptable prodrug, salt, polymorph,
co-crystal, or solvate of A-001. In certain of these embodiments, the one or
more sPLA2 inhibitors comprise A-002, a prodrug of A-001. In certain
embodiments, the one or more compounds used in the treatment of CVD
comprise one or more statins or statin combination drugs. In certain of these
embodiments, the one or more statins are selected from the group consisting
of atorvastatin, simvastatin, rosuvastatin, lovastatin, pravastatin,
cerivastatin,
fluvastatin, mevastatin, pitavastatin, and various salts, solvates,
stereoisomers, and prodrug derivatives thereof. In certain of these
embodiments, the statin combination drugs are selected from the group
12
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
consisting of atorvastatin plus ezetimibe, atorvastatin plus amiodipine,
atorvastatin plus CP-529414, atorvastatin plus APA-01, simvastatin plus
ezetimibe, simvastatin plus extended release niacin, simvastatin plus MK-
0524A, lovastatin plus extended release niacin, rosuvastatin plus fenofibrate,
pravastatin plus fenofibrate, and statin plus TAK-457. In certain
embodiments, the one or more compounds used in the treatment of CVD
comprise one or more non-statin compounds selected from the group
consisting of bile acid sequestrants, fibrates, niacin or niacin derivatives,
cholesterol absorption inhibitors, cholesteryl ester transfer protein (CETP)
inhibitors, microsomal triglyceride transfer protein (MTP) inhibitors,
squalene
synthase inhibitors, ACE inhibitors, angiotensin ll receptor antagonists, beta-
adrenergic blockers, calcium channel blockers, and antithrombotics. In
certain embodiments, the one or more sPLA2 inhibitors and the one or more
compounds used in the treatment of CVD may be administered
simultaneously. In certain of these embodiments, the one or more sPLA2
inhibitors and the one or more compounds used in the treatment of CVD may
be administered in a single formulation, while in other embodiments the
compounds may be administered simultaneously in two or more formulations.
In each of these embodiments, the formulation(s) may further comprise one or
more pharmaceutically acceptable carriers. In other embodiments, the one or
more sPLA2 inhibitors and the one or more compounds used in the treatment
of CVD may be administered sequentially. In certain embodiments, the one
or more sPLA2 inhibitors may be administered to the subject twice or more per
day, and in certain of these embodiments the one or more sPLA2 inhibitors
are administered twice per day. In other embodiments, the one or more
13
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
sPLA2 inhibitors may be administered to the subject on a once a day basis. In
certain embodiments, the one or more sPLA2 inhibitors may be administered
at a dosage of about 50 to about 500 mg.
[0013] In certain embodiments, methods are provided for increasing HDL
levels in a subject in need thereof by administering a therapeutically
effective
amount of one or more sPLA2 inhibitors and a therapeutically effective amount
of one or more compounds used in the treatment of CVD. In certain
embodiments, the one or more sPLA2 inhibitors comprise A-001 or a
pharmaceutically acceptable prodrug, salt, polymorph, co-crystal, or solvate
of
A-001. In certain of these embodiments, the one or more sPLA2 inhibitors
comprise A-002, a prodrug of A-001. In certain of these embodiments, the
one or more compounds used in the treatment of CVD comprise one or more
statins or statin combination drugs. In certain of these embodiments, the one
or more statins are selected from the group consisting of atorvastatin,
simvastatin, rosuvastatin, lovastatin, pravastatin, cerivastatin, fluvastatin,
mevastatin, pitavastatin, and various salts, solvates, stereoisomers, and
prodrug derivatives thereof. In certain of these embodiments, the statin
combination drugs are selected from the group consisting of atorvastatin plus
ezetimibe, atorvastatin plus amlodipine, atorvastatin plus CP-529414,
atorvastatin plus APA-01, simvastatin plus ezetimibe, simvastatin plus
extended release niacin, simvastatin plus MK-0524A, lovastatin plus extended
release niacin, rosuvastatin plus fenofibrate, pravastatin plus fenofibrate,
and
statin plus TAK-457. In certain embodiments, the one or more compounds
used in the treatment of CVD comprise one or more non-statin compounds
selected from the group consisting of bile acid sequestrants, fibrates, niacin
or
14
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
niacin derivatives, cholesterol absorption inhibitors, cholesteryl ester
transfer
protein (CETP) inhibitors, microsomal triglyceride transfer protein (MTP)
inhibitors, squalene synthase inhibitors, ACE inhibitors, angiotensin li
receptor
antagonists, beta-adrenergic blockers, calcium channel blockers, and
antithrombotics. In certain embodiments, administration of one or more sPLA2
inhibitors and one or more compounds used in the treatment of CVD results in
an improvement in HDL/LDL ratio. In certain embodiments, improvement in
HDL/LDL ratio is further accomplished by a reduction in LDL levels. In certain
embodiments, the one or more sPLA2 inhibitors and the one or more
compounds used in the treatnient of CVD may be administered
simultaneously. In certain of these embodiments, the one or more sPLA2
inhibitors and the one or more compounds used in the treatment of CVD may
be administered in a single formulation, while in other embodiments the
compounds may be administered simultaneously in two or more formulations.
In each of these embodiments, the formulation(s) may further comprise one or
more pharmaceutically acceptable carriers. In other embodiments, the one or
more sPLA2 inhibitors and the one or more compounds used in the treatment
of CVD may be administered sequentially. In certain embodiments, the one
or more sPLA2 inhibitors may be administered to the subject twice or more per
day, and in certain of these embodiments the one or more sPLA2 inhibitors
are administered twice per day. In other embodiments, the one or more
sPLA2 inhibitors may be administered to the subject on a once a day basis. In
certain embodiments, the one or more sPLA2 inhibitors may be administered
at a dosage of about 50 to about 500 mg.
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
[0014] In certain embodiments, methods are provided for treating CVD or a
condition associated with CVD in a subject in need thereof by administering a
therapeutically effective amount of one or more sPLA2 inhibitors and one or
more compounds used in the treatment of CVD. In certain embodiments, the
one or more sPLA2 inhibitors comprise A-001 or a pharmaceutically
acceptable prodrug, salt, polymorph, co-crystal, or solvate of A-001. In
certain of these embodiments, the one or more sPLA2 inhibitors comprise A-
002, a prodrug of A-001. In certain embodiments, the one or more
compounds used in the treatment of CVD comprise one or more statins or
statin combination drugs. In certain of these embodiments, the one or more
statins are selected from the group consisting of atorvastatin, simvastatin,
rosuvastatin, lovastatin, pravastatin, cerivastatin, fluvastatin, mevastatin,
pitavastatin, and various salts, solvates, stereoisomers, and prodrug
derivatives thereof. In certain of these embodiments, the statin combination
drugs are selected from the group consisting of atorvastatin plus ezetimibe,
atorvastatin plus amlodipine, atorvastatin plus CP-529414, atorvastatin plus
APA-01, simvastatin plus ezetimibe, simvastatin plus extended release niacin,
simvastatin plus MK-0524A, lovastatin plus extended release niacin,
rosuvastatin plus fenofibrate, pravastatin plus fenofibrate, and statin plus
TAK-457. In certain embodiments, the one or more compounds used in the
treatment of CVD comprise one or more non-statin compounds selected from
the group consisting of bile acid sequestrants, fibrates, niacin or niacin
derivatives, cholesterol absorption inhibitors, cholesteryl ester transfer
protein
(CETP) inhibitors, microsomal triglyceride transfer protein (MTP) inhibitors,
squalene synthase inhibitors, ACE inhibitors, angiotensin 11 receptor
16
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
antagonists, beta-adrenergic blockers, calcium channel blockers, and
antithrombotics. In certain embodiments, administration of one or more sPLA2
inhibitors results in a decrease in levels of one or more inflammatory
markers.
In certain of these embodiments, the inflammatory markers may include, but
are not limited to, sPLA2, CRP, and/or IL-6. In certain embodiments, CVD or
a condition associated with CVD includes, but is not limited to,
atherosclerosis, metabolic syndrome, coronary artery disease, coronary heart
disease, cerebrovascular disease, peripheral vascular disease, and/or
conditions associated with coronary artery disease, coronary heart disease,
cerebrovascular disease, or peripheral vascular disease. In certain
embodiments, the one or more sPLA2 inhibitors and the one or more
compounds used in the treatnient of CVD may be administered
simultaneously. In certain of these embodiments, the one or more sPLA2
inhibitors and the one or more compounds used in the treatrnent of CVD may
be administered in a single formulation, while in other embodiments the
compounds may be administered simultaneously in two or more formulations.
In each of these embodiments, the formulation(s) may further comprise one or
more pharmaceutically acceptable carriers. In other embodiments, the one or
more sPLA2 inhibitors and the one or more compounds used in the treatment
of CVD may be administered sequentially. In certain embodiments, the one
or more sPLA2 inhibitors may be administered to the subject twice or more per
day, and in certain of these embodiments the one or more sPLA2 inhibitors
are administered twice per day. In other embodiments, the one or more
sPLA2 inhibitors may be administered to the subject on a once a day basis. In
17
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
certain embodiments, the one or more sPLA2 inhibitors may be administered
at a dosage of about 50 to about 500 mg.
[0015] In certain embodiments, methods are provided for treating metabolic
syndrome in a subject in need thereof by administering a therapeutically
effective amount of one or more sPLA2 inhibitors and one or more compounds
used in the treatment of CVD. In certain embodiments, the one or more
sPLA2 inhibitors comprise A-001 or a pharmaceutically acceptable prodrug,
salt, polymorph, co-crystal, or solvate of A-001. In certain of these
embodiments, the one or more sPLA2 inhibitors comprise A-002, a prodrug of
A-001. In certain embodiments, the one or more compounds used in the
treatment of CVD comprise one or more statins or statin combination drugs.
In certain of these embodiments, the one or more statins are selected from
the group consisting of atorvastatin, simvastatin, rosuvastatin, lovastatin,
pravastatin, cerivastatin, fluvastatin, mevastatin, pitavastatin, and various
salts, solvates, stereoisomers, and prodrug derivatives thereof. In certain of
these embodiments, the statin combination drugs are selected from the group
consisting of atorvastatin plus ezetimibe, atorvastatin plus amlodipine,
atorvastatin plus CP-529414, atorvastatin plus APA-01, simvastatin plus
ezetimibe, simvastatin plus extended release niacin, simvastatin plus MK-
0524A, lovastatin plus extended release niacin, rosuvastatin plus fenofibrate,
pravastatin plus fenofibrate, and statin plus TAK-457. In certain
embodiments, the one or more compounds used in the treatment of CVD
comprise one or more non-statin compounds selected from the group
consisting of bile acid sequestrants, fibrates, niacin or niacin derivatives,
cholesterol absorption inhibitors, cholesteryl ester transfer protein (CETP)
18
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
inhibitors, microsomal triglyceride transfer protein (MTP) inhibitors,
squalene
synthase inhibitors, ACE inhibitors, angiotensin II receptor antagonists, beta-
adrenergic blockers, calcium channel blockers, and antithrombotics. In
certain embodiments, administration of one or more sPLA2 inhibitors results in
a decrease in levels of one or more inflammatory markers. In certain of these
embodiments, the inflammatory markers may include, but are not limited to,
sPLA2, CRP, and/or IL-G. In certain embodiments, the one or more sPLA2
inhibitors and the one or more compounds used in the treatment of CVD may
be administered simultaneously. In certain of these embodiments, the one or
more sPLA2 inhibitors and the one or more compounds used in the treatment
of CVD may be administered in a single formulation, while in other
embodiments the compounds may be administered simultaneously in two or
more formulations. In each of these embodiments, the formulation(s) may
further comprise one or more pharmaceutically acceptable carriers. In other
embodiments, the one or more sPLA2 inhibitors and the one or more
compounds used in the treatment of CVD may be administered sequentially.
In certain embodiments, the one or more sPLA2 inhibitors may be
administered to the subject twice or more per day, and in certain of these
embodiments the one or more sPLA2 inhibitors are administered twice per
day. In other embodiments, the one or more sPLA2 inhibitors may be
administered to the subject on a once a day basis. In certain embodiments,
the one or more sPLA2 inhibitors may be administered at a dosage of about
50 to about 500 mg.
[0016] In certain embodiments, methods are provided for increasing the
effectiveness of a compound used in the treatment of CVD in a subject by
19
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
administering one or more sPLA2 inhibitors to said subject. In certain
embodiments, the one or more sPLA2 inhibitors comprise A-001 or a
pharmaceutically acceptable prodrug, salt, polymorph, co-crystal, or solvate
of
A-001. In certain of these embodiments, the one or more sPLA2 inhibitors
comprise A-002, a prodrug of A-001. In certain embodiments, the compound
used in the treatment of CVD may be a statin or statin combination drug. In
certain of these embodiments, the statin may be selected from the group
consisting of atorvastatin, simvastatin, rosuvastatin, lovastatin,
pravastatin,
cerivastatin, fluvastatin, mevastatin, pitavastatin, and various salts,
solvates,
stereoisomers, and prodrug derivatives thereof. In certain of these
embodiments, the statin combination drug may be selected from the group
consisting of atorvastatin plus ezetimibe, atorvastatin plus amlodipine,
atorvastatin plus CP-529414, atorvastatin plus APA-01, simvastatin plus
ezetimibe, simvastatin plus extended release niacin, simvastatin plus MK-
0524A, lovastatin plus extended release niacin, rosuvastatin plus fenofibrate,
pravastatin plus fenofibrate, and statin plus TAK-457. In certain
embodiments, the compound used in the treatment of CVD may be a non-
statin compound selected from the group consisting of bile acid sequestrants,
fibrates, niacin or niacin derivatives, cholesterol absorption inhibitors,
cholesteryl ester transfer protein (CETP) inhibitors, microsomal triglyceride
transfer protein (MTP) inhibitors, squalene synthase inhibitors, ACE
inhibitors,
angiotensin il receptor antagonists, beta-adrenergic blockers, calcium channel
blockers, and antithrombotics. In certain embodiments, the one or more
sPLA2 inhibitors may be administered simultaneously with the compound used
in the treatment of CVD. In certain of these embodiments, the one or more
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
sPLA2 inhibitors and the compound used in the treatment of CVD may be
administered in a single formulation, while in other embodiments the
compounds may be administered simultaneously in two or more formulations.
In each of these embodiments, the formulation(s) may further comprise one or
more pharmaceutically acceptable carriers. In other embodiments, the one or
more sPLA2 inhibitors and the compound used in the treatment of CVD may
be administered sequentially. In certain embodiments, the one or more sPLA2
inhibitors may be administered to the subject twice or more per day, and in
certain of these embodiments the one or more sPLA2 inhibitors are
administered twice per day. In other embodiments, the one or more sPLA2
inhibitors may be administered to the subject on a once a day basis. In
certain embodiments, the one or more sPLA2 inhibitors may be administered
at a dosage of about 50 to about 500 mg.
[0097] In certain embodiments, the use of one or more sPLA2 inhibitors for
preparation of a medicament for treating dyslipidemia, treating CVD and
conditions associated with CVD, lowering cholesterol levels, lowering
triglyceride levels, increasing HDL levels, and improving HDL/LDL ratios in a
subject is provided. In certain embodiments, the one or more sPLA2 inhibitors
comprise A-001 or a pharmaceutically acceptable prodrug, salt, polymorph,
co-crystal, or solvate of A-001. In certain of these embodiments, the one or
more sPLA2 inhibitors comprise A-002, a prodrug of A-001. In certain
embodiments, the medicament further coniprises one or more
pharmaceutically acceptable carriers.
[0018] In certain embodiments, the use of one or more sPLA2 inhibitors and
one or more compounds used in the treatment of CVD for preparation of a
21
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
medicament for treating dyslipidemia, treating CVD and conditions associated
with CVD, lowering cholesterol levels, lowering triglyceride levels,
increasing
HDL levels, and/or improving HDL/LDL ratios in a subject is provided. In
certain embodiments, the one or more sPLA2 inhibitors comprise A-001 or a
pharmaceutically acceptable prodrug, salt, polymorph, co-crystal, or solvate
of
A-001. In certain of these embodiments, the one or more sPLA2 inhibitors
comprise A-002, a prodrug of A-001. In certain embodiments, the one or
more compounds used in the treatrnent of CVD comprise one or more statins
or statin combination drugs. In certain of these embodiments, the one or
more statins are selected from the group consisting of atorvastatin,
simvastatin, rosuvastatin, lovastatin, pravastatin, cerivastatin, fluvastatin,
mevastatin, pitavastatin, and various salts, solvates, stereoisomers, and
prodrug derivatives thereof. In certain of these embodiments, the statin
combination drugs are selected from the group consisting of atorvastatin plus
ezetimibe, atorvastatin plus amiodipine, atorvastatin plus CP-529414,
atorvastatin plus APA-01, simvastatin plus ezetimibe, simvastatin plus
extended release niacin, simvastatin plus MK-0524A, lovastatin plus extended
release niacin, rosuvastatin plus fenofibrate, pravastatin plus fenofibrate,
and
statin plus TAK-457. In certain embodiments, the one or more compounds
used in the treatment of CVD comprise one or more non-statin compounds
selected from the group consisting of bile acid sequestrants, fibrates, niacin
or
niacin derivatives, cholesterol absorption inhibitors, cholesteryl ester
transfer
protein (CETP) inhibitors, microsomal triglyceride transfer protein (MTP)
inhibitors, squalene synthase inhibitors, ACE inhibitors, angiotensin 11
receptor
antagonists, beta-adrenergic blockers, calcium channel blockers, and
22
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
antithrombotics. In certain embodiments, the medicament further comprises
one or more pharmaceutically acceptable carriers.
[0019] In addition to the exemplary embodiments described above, further
embodiments and aspects will become apparent by reference to the drawings
and by study of the following descriptions.
BRIEF DESCRIPTION OF DRAWINGS
[0020] Figure 1: Effect of A-002 administration on body weight in mice.
ApoE-'- mice were administered vehicle only, 30 mg/kg A-002, or 90 mg/kg A-
002 twice daily over 16 weeks. Body weight was measured once a week.
[0021] Figure 2: Effect of A-002 administration on plasma total cholesterol
levels in mice. ApoE-'- mice were administered vehicle only, 30 mg/kg A-002,
or 90 mg/kg A-002 twice daily over 16 weeks. Total plasma cholesterol levels
were measured at 0, 4, 8, 12, and 16 weeks.
[0022] Figure 3: Effect of A-002 administration on atherosclerotic plaque
coverage in mice. ApoE-'- mice were administered vehicle only, 30 mg/kg A-
002, or 90 mg/kg A-002 twice daily over 16 weeks. At 16 weeks, aortic
plaque coverage was measured.
[0023] Figure 4: Effect of A-002 administration on angiotensin II-mediated
atherosclerotic plaque formation and aortic aneurysm. ApoE-'- mice were
administered saline plus water, angiotensin II in water vehicle, angiotensin
11
in acacia vehicle, or angiotensin II in acacia vehicle plus 30 mg/kg A-002. At
4 weeks, aortic plaque coverage was measured.
[0024] Figure 5: En face lesions in A-002 and/or statin dosage groups.
ApoE-'- mice on a high fat diet were administered various dosages of A-002,
statin, or A-002 plus statin over twelve weeks, and en face lesion content was
23
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
measured using digital imaging methods. Lesion size is expressed as percent
coverage over entire tissue sample. A: low dose A-002; B: high dose A-002;
C: statin; D: low dose A-002 plus statin; E: high dose A-002 plus statin; and
F:
vehicle only.
[0025] Figure 6: Total plasma cholesterol levels in A-002 and/or statin
dosage groups. ApoE"/- mice on a high fat diet were administered various
dosages of A-002, statin, or A-002 plus statin over twelve weeks, and total
cholesterol levels were measured. A: low dose A-002; B: high dose A-002; C.
statin; D: low dose A-002 plus statin; E: high dose A-002 plus statin; and F:
vehicle only.
[0026] Figure 7: HDL levels in A-002 and/or statin dosage groups. ApoE-'-
mice on a high fat diet were administered various dosages of A-002, statin, or
A-002 plus statin over twelve weeks, and HDL levels were measured. A: low
dose A-002; B: high dose A-002; C: statin; D: low dose A-002 plus statin; E.
high dose A-002 plus statin; and F: vehicle only.
[0027] Figure 8: A-002/simvastatin combination tablet preparation protocol.
[0028] Figure 9: HPLC profiles. A. Simvastatin. B. A-002. C. A-
002/simvastatin combination tablet #19. D. A-002/simvastatin combination
tablet #27.
DETAILED DESCRIPTION
[0029] The following description of the invention is merely intended to
illustrate various embodiments of the invention. As such, the specific
modifications discussed are not to be construed as limitations on the scope of
the invention. It will be apparent to one skilled in the art that various
equivalents, changes, and modifications may be made without departing from
24
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
the scope of the invention, and it is understood that such equivalent
embodiments are to be included herein.
Abbreviations:
[0030] AA, arachidonic acid; ACE, angiotensin converting enzyme; Ang,
angiotensin; ApoB, apolipoprotein B; ARB, Angiotensin Receptor Blocker;
BID, twice daily; BMI, body mass index; CAD, coronary artery disease; CETP,
cholesteryl ester transfer protein; cfm, cubic feet per minute; CHD, coronary
heart disease; CRP, C-reactive protein; CVD, cardiovascular disease; ECG,
electrocardiogram; ERN, extended release niacin; HDL, high density
lipoprotein; HMG-CoA, hydroxymethyl glutaryl coenzyme A; HPLC, high
performance liquid chromatography; ICAM-1, intercellular adhesion molecule
1; IDL, intermediate density lipoprotein; IL, interleukin (e.g., IL-6, 1L-8);
ITT,
intent to treat; LDL, low density lipoprotein; LPA, lysophosphatidic acid; MCP-
1, monocyte chemotactic protein-1; MI, myocardial infarction; MIP-1a,
macrophage inflammatory protein 1 alpha; MTP, microsomal triglyceride
transfer protein; PAD, peripheral artery disease; PAF, platelet activating
factor; PLA2, phospholipase A2; QD, once daily; sPLA2, secretory
phospholipase A2; TEAE, treatment-emergent adverse event; TG, triglyceride;
TIA, transient ischemic attack; TNFa, tumor necrosis factor alpha; VCAM-1,
vascular cell adhesion molecule 1; VLDL, very low density lipoprotein.
[0031] The terms "treat," "treating," or "treatment" as used herein with
regards to a condition refers to preventing the condition, slowing the onset
or
rate of development of the condition, reducing the risk of developing the
condition, preventing or delaying the development of symptoms associated
with the condition, reducing or ending symptoms associated with the
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
condition, generating a complete or partial regression of the condition, or
some combination thereof. For example, with regard to atherosclerosis,
"treatment" may refer to a decrease in the likelihood of developing
atherosclerotic plaque deposits, a decrease in the rate of development of
deposits, a decrease in the number or size of existing deposits, or improved
plaque stability. Likewise, "treatment" with regard to dyslipidemia may refer
to
a decrease in lipid levels, cholesterol levels, and/or triglyceride (TG)
levels.
[0032] The term "subject" as used herein refers to any mammal, preferably a
human.
[0033] In certain embodiments, a "subject in need thereof' refers to a subject
diagnosed with CVD or exhibiting one or more conditions associated with
CVD, a subject who has been diagnosed with or exhibited one or more
conditions associated with CVD in the past, or a subject who has been
deemed at risk of developing CVD or one or more conditions associated with
CVD in the future due to hereditary or environmental factors. "Cardiovascular
disease" or "CVD" as used herein includes, for example, atherosclerosis,
including coronary artery atherosclerosis and carotid artery atherosclerosis,
coronary artery disease (CAD), coronary heart disease (CHD), conditions
associated with CAD and CHD, cerebrovascular disease and conditions
associated with cerebrovascular disease, peripheral vascular disease and
conditions associated with peripheral vascular disease, aneurysm, vasculitis,
venous thrombosis, diabetes mellitus, and metabolic syndrome. "Conditions
associated with CAD and CHD" as used herein include, for example, angina
and myocardial infarction (Ml; heart attack). "Conditions associated with
cerebrovascular disease" as used herein include, for example, transient
26
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
ischemic attack (T1A) and stroke. "Conditions associated with peripheral
vascular disease" as used herein include, for example, claudication.
"Conditions associated with CVD" as used herein include, for example,
dyslipidemia, such as for example hyperlipidemia (elevated lipid levels),
hypercholesterolemia (elevated cholesterol levels), and hypertriglyceridemia
(elevated TG levels), elevated glucose levels, low HDL/LDL ratio, and
hypertension. Therefore, in certain embodiments, a subject in need thereof
may be a subject exhibiting dyslipidemia or a subject that has exhibited
dyslipidemia in the past or has been deemed at risk for developing
dyslipidemia in the future. In certain of these embodiments, the subject may
exhibit elevated cholesterol levels, or may have exhibited elevated
cholesterol
levels in the past or been deemed at risk for developing elevated cholesterol
levels in the future. Likewise, in certain of these embodiments, the subject
may exhibit elevated triglyceride levels, or may have exhibited elevated
triglyceride levels in the past or been deemed at risk for developing elevated
triglyceride levels in the future. In certain embodiments, a subject in need
thereof may have a condition associated with inflammation, may have been
diagnosed with such a condition in the past, or may have been deemed at risk
for developing such a condition in the future. In addition to atherosclerosis
and certain other forms of CVD, conditions associated with inflammation
include, for example, multiple sclerosis (Cunningham 2006), Alzheimer's
disease (Moses 2006), sickle cell (Styles 1996), rheumatoid arthritis, and
osteoarthritis (Jamal 1998). In these embodiments, a subject in need thereof
may exhibit elevated sPLA2 levels, may have exhibited elevated sPLA2 levels
in the past, or may have been deemed at risk for developing elevated sPLA2
27
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
levels. In other embodiments, a subject in need thereof may exhibit sPLA2
levels falling within a normal range. In certain embodiments, a subject in
need thereof may exhibit elevated levels of one or more additional markers
associated with inflammation, including but not limited to CRP, IL-6, MCP-1,
TNFa, IL-8, ICAM-1, VCAM-1, and MIP-la.
[0034] The term "cholesterol level" as used herein refers to blood cholesterol
level, serum cholesterol level, plasma cholesterol level, or cholesterol level
from another biological fluid. A decrease in cholesterol levels as used herein
may refer to a decrease in total cholesterol levels or a decrease in one or
more of total cholesterol, non-HDL cholesterol, LDL, VLDL, and/or IDL levels.
A decrease in LDL as used herein may refer to a decrease in total LDL, a
decrease in LDL particles, a decrease in small LDL particles, a decrease in
oxidized LDL levels, and/or a decrease in ApoB levels. A decrease in VLDL
as used herein may refer to a decrease in total VLDL or to a decrease in the
level of one or more of VLDL subparticles V1 to V6. An improvement in
HDL/LDL ratio as used herein refers to any increase in the ratio of HDL to
LDL, and may be accomplished by decreasing LDL levels, increasing HDL
levels, or some combination thereof. An increase in LDL particle size as used
herein refers to an increase in mean particle size.
[0035] The term "elevated cholesterol level" as used herein refers to a
cholesterol level that is above an accepted normal threshold level, such as
those promulgated by the National Heart Lung and Blood Institute (NHLBI)
National Cholesterol Education Program (NCEP). The accepted norrnal
threshold cholesterol level may vary from subject to subject based on various
risk factors, such as for example a prior history of CVD. In certain
28
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
embodiments, a subject exhibiting elevated cholesterol levels may have a
blood LDL level greater than or equal to 70 mg/dl. In certain of these
embodiments, a subject exhibiting elevated cholesterol levels may have a
blood LDL greater than or equal to 100 mg/dl, in other embodiments greater
than or equal to 130 mg/dl, in other embodiments greater than or equal to 160
mg/dl, and in still other embodiments greater than or equal to 190 mg/di. In
certain embodiments, a subject exhibiting elevated cholesterol levels may
have a blood total cholesterol level greater than or equal to 200 mg/dl. In
certain of these embodiments, a subject exhibiting elevated cholesterol levels
may have a blood total cholesterol greater than or equal to 240 mg/dl.
[0036] The term "triglyceride level" as used herein refers to blood
triglyceride
level, serum triglyceride level, plasma triglyceride level, or triglyceride
level
from another biological fluid. The term "elevated triglyceride level" as used
herein refers to a triglyceride level that is above an accepted normal
threshold
level. The accepted normal threshold triglyceride level may vary from subject
to subject based on various risk factors, such as for example a prior history
of
CVD. In certain embodiments, a subject exhibiting elevated triglyceride levels
may have a blood triglyceride level greater than or equal to 150 mg/dI. In
certain of these embodiments, a subject exhibiting elevated triglyceride
levels
may have a blood triglyceride level greater than or equal to 200 mg/dl, in
other
embodiments greater than or equal to 300 mg/dl, in other embodiments
greater than or equal to 400 mg/dl, and in still other embodiments greater
than
or equal to 500 mg/dl.
29
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
[0037] The term "statin" as used herein refers to any compound that inhibits
HMG-CoA reductase, an enzyme that catalyzes the conversion of HMG-CoA
to mevalonate.
[0038] The term "sPLA2 inhibitor" as used herein refers to any compound or
prodrug thereof that inhibits the activity of sPLA2.
[0039] A"therapeutically effective amount" of a composition as used herein
is an amount of a composition that produces a desired therapeutic effect in a
subject, such as treating a target condition. The precise therapeutically
effective amount is an amount of the composition that will yield the most
effective results in terms of therapeutic efficacy in a given subject. This
amount will vary depending upon a variety of factors, including but not
limited
to the characteristics of the therapeutic composition (including, e.g.,
activity,
pharmacokinetics, pharmacodynamics, and bioavailability), the physiological
condition of the subject (including, e.g., age, body weight, sex, disease type
and stage, medical history, general physical condition, responsiveness to a
given dosage, and other present medications), the nature of the
pharmaceutically acceptable carrier or carriers in the composition, and the
route of administration. One skilled in the clinical and pharmacological arts
will be able to determine a therapeutically effective amount through routine
experimentation, namely by monitoring a subject's response to administration
of a composition and adjusting the dosage accordingly. For additional
guidance, see, e.g., Remington: The Science and Practice of Pharmacy, 21s'
Edition, Univ. of Sciences in Philadelphia (USIP), Lippincott Williams &
Wilkins, Philadelphia, PA, 2005, and Goodman & Gilman's The
Pharmacological Basis of Therapeutics, 11th Edition, McGraw-Hill, New York,
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
NY, 2006, the entire disclosures of which are incorporated by reference
herein.
[0040] A"pharmaceutically acceptable carrier" as used herein refers to a
pharmaceutically acceptable material, composition, or vehicle that is involved
in carrying or transporting a compound of interest from one tissue, organ, or
portion of the body to another tissue, organ, or portion of the body. Such a
carrier may comprise, for example, a liquid, solid, or semi-solid filler,
solvent,
surfactant, diluent, excipient, adjuvant, binder, buffer, dissolution aid,
solvent,
encapsulating material, sequestering agent, dispersing agent, preservative,
lubricant, disintegrant, thickener, emulsifier, antimicrobial agent,
antioxidant,
stabilizing agent, coloring agent, or some combination thereof. Each
component of the carrier must be "pharmaceutically acceptable" in that it must
be compatible with the other ingredients of the composition and must be
suitable for contact with any tissue, organ, or portion of the body that it
may
encounter, meaning that it must not carry a risk of toxicity, irritation,
allergic
response, immunogenicity, or any other complication that excessively
outweighs its therapeutic benefits. Examples of pharmaceutically acceptable
carriers for use in the presently disclosed pharmaceutical compositions
include, but are not limited to, diluents such as microcrystaHine cellulose or
lactose (e.g., anhydrous lactose, lactose fast flo), binders such as gelatin,
polyethylene glycol, wax, microcrystalline cellulose, synthetic gums such as
polyvinylpyrrolidone, or cellulosic polymers such as hydroxypropyl cellulose
(e.g., hydroxypropyl methylcellulose (HPMC)), lubricants such as magnesium
stearate, calcium stearate, stearic acid, or microcrystalline cellulose,
disintegrants such as starches, cross-linked polymers, or celluloses (e.g.,
31
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
croscarmellose sodium (CCNa), fillers such as silicon dioxide, titanium
dioxide, microcrystalline cellulose, or powdered cellulose, surfactants or
emulsifiers such as polysorbates (e.g., Polysorbate 20, 40, 60, or 80; Span
20, 40, 60, 65, or 80), antioxidant agents such as butylated hydroxyanisole
(BHA), butylated hydroxytoluene (BHT), propyl gailate, or ascorbic acid
(either
free acid or salt forms thereof), buffers such as phosphate or citrate
buffers,
sequestering agents such as ethylenediaminetetraacetic acid (EDTA),
ethylene glycol tetraacetic acid (EGTA), or edetate disodium, dispersing
agents such as sodium carboxymethylcelluose, hydroxypropyl
methylcellulose, povidone, or polyvinylpyrrolidone, dissolution aids such as
calcium carbonate, and excipients such as water, saline, dextrose, glycerol,
or
ethanol, citric acid, calcium metabisulfite, lactic acid, malic acid, succinic
acid,
or tartaric acid.
[0041] Metabolic syndrome is a disorder characterized by a group of
metabolic risk factors. These factors include, for example, dyslipidemia,
abdominal obesity, elevated blood pressure (hypertension), insulin resistance
or glucose intolerance, prothrombotic state, and proinflammatory state.
Subjects are generally classified as having metabolic syndrome if they meet
three of the five following criteria: 1) abdominal obesity (waist
circumference
>35 inches in women, >40 inches in men); 2) low HDL levels (<50 mg/dL in
women, <40 mg/dL in men); 3) high blood pressure (>130/85 mm Hg) or
current treatment with antihypertensive medication; 4) hypertriglyceridemia
(TG levels >_150 mgldL); and 5) impaired fasting glucose (blood glucose levels
of >110 mg/dL). Metabolic syndrome is associated with elevated levels of
various inflammatory markers, such as CRP or IL-6. Subjects with metabolic
32
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
syndrome are at increased risk of developing CAD, CHD, conditions
associated with CAD and CHD, and type 2 diabetes,
[0042] Over time, hypercholesterolemia and hypertriglyceridemia can lead to
the development of atheromatous plaques on the inner arterial linings via the
process of atherogenesis, which in turn results in atherosclerosis.
Atherosclerosis leads to significantly reduced blood flow through the
arteries,
which in turn leads to the development of CAD, CHD, and conditions
associated with CAD and CHD.
[0043] Reduction of cholesterol levels, particularly LDL levels, and/or
reduction of TG levels have been shown to delay the onset and decrease the
progression of atherosclerosis, thereby reducing the risk of developing CAD
and CHD. In certain cases, adequate cholesterol and/or TG reduction may be
accomplished through diet adjustment. In other cases, however, adequate
reduction requires the administration of one or more compounds used in the
treatment of CVD. Compounds used in the treatment of CVD include
compounds for lowering cholesterol levels and/or increasing HDL levels, such
as for example statins, bile acid sequestrants such as cholestyramine resin
(Questran0. Prevalite0), colestipol hydrochloride (Colestid0), or colesevelam
hydrochloride (WelCholO. Cholestagel0), fibrates such as bezafibrate
(Bezalip0), ciprofibrate (Modalim0), clofibrate, gemfibrozil (LopidO), or
fenofibrate (Antara0, TriCorO, ABT-335), niacin or niacin derivatives such as
xanthinol niacinate, niacin immediate-release (Niacor0), extended release
forms of niacin (ERN) (Niaspan0, Niaspan MF, or Niaspan CF), or extended
release niacin combinations (e.g., extended release niacin plus the DP-1
antagonist laropiprant (MK-0524), combination known as MK-0524A and
33
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
marketed as Cordaptive ), cholesterol absorption inhibitors such as ezetimibe
(Zetia(@), AVE 5530, or MD-0727, cholesteryl ester transfer protein (CETP)
inhibitors such as JTT-705/R04607381 (R1658), CP-529414 (Torcetrapib ),
or MK-0859, microsomal triglyceride transfer protein (MTP) inhibitors such as
AEGR-733 and AEGR-733 combinations (e.g., AEGR-733 plus ezetimibe),
squalene synthase inhibitors such as lapaquistat acetate (TAK-475) and
lapaquistat acetate combinations (e.g., TAK-475 plus one or more statins),
and other miscellaneous compounds such as dextrothyroxine, ISIS 301012,
cardioprotectants such as MC-1 antibody, glycoprotein Ilb/Illa inhibitors such
as tirofiban hydrochloride (Aggrastat ), TG100-115, AEGR 773, AEGR 427,
stanols, or sterols. In addition to compounds that lower cholesterol levels or
increase HDL levels, compounds used in the treatment of CVD include, for
example, ACE inhibitors such as lisinopril, captopril, enalapril, nitrosated
ACE
inhibitors, or ACE inhibitor combinations (e.g., lisinopril plus MC-1
antibody,
combination referred to as MC-4232), angiotensin II receptor antagonists,
nitrosated angiotensin II receptor antagonists, or angiotensin !I receptor
antagonist combinations (e.g., Angiotensin Receptor Blocker (ARB) plus MC-
1, combination referred to as MC-4262), beta-adrenergic blockers or
nitrosated beta-adrenergic blockers, calcium channel blockers, or
antithrombotics such as aspirin or nitrosated aspirin.
[0044] Among the most well-known and commonly used compounds used in
the treatment of CVD are statins. Statins are compounds that inhibit HMG-
CoA reductase from catalyzing the conversion of HMG-CoA to mevalonate, a
rate-limiting step in the cholesterol biosynthetic pathway. As such, statins
function as potent lipid lowering agent. Statin administration significantly
34
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
decreases blood LDL levels, and moderately decreases blood TG levels. In
addition, it has been proposed that statins may prevent CVD by improving
endothelial function, modulating inflammatory responses, maintaining plaque
stability, and preventing thrombus formation. Examples of statins that may be
used in conjunction with the compositions and methods disclosed herein
include, but are not limited to, atorvastatin or atorvastatin calcium
(marketed
as Lipitor0 or Torvast O; see, e.g., U.S. Patent Nos. 4,681,893 or 5,273,995)
and atorvastatin combinations (e.g., atorvastatin plus amlodipine (marketed
as Norvasc0), combination marketed as Caduet(@, see, e.g., U.S. Patent No.
6,455,574; atorvastatin plus CP-529414 (marketed as Torcetrapib0);
atorvastatin plus APA-01; atorvastatin plus ezetimibe), cerivastatin (marketed
as Lipobay0 or Baycol0), fluvastatin (marketed as Lescol0; U.S. Patent No.
4,739,073). lovastatin (marketed as Mevacor0 or Altocor0; see, e.g., U.S.
Patent No. 4,231,938). lovastatin combinations (e.g., lovastatin plus
Niaspan0, combination marketed as Advicor0), mevastatin, pitavastatin
(marketed as Livalo0 or Pitava0), pravastatin (marketed as Pravachol0,
Mevalotin0, Selektine(@, or Lipostat0; see, e.g., U.S. Patent No. 4,346,227),
pravastatin combinations (e.g., pravastatin plus fenofibrate), rosuvastatin
(marketed as Crestor0), rosuvastatin combinations (e.g., rosuvastatin plus
TriCorO), simvastatin (marketed as ZocorO or LipexO; see, e.g., U.S. Patent
Nos. 4,444,784; 4,916,239; and 4,820,850), and simvastatin combinations
(e.g., simvastatin plus ezetimibe, combination marketed as Vytorin0, see,
e.g., U.S. Patent No. 7,229,982; simvastatin plus Niaspan0, combination
marketed as Simcor0; simvastatin plus MK-0524A, combination referred to as
MK-0524B), as well as various pharmaceutically acceptable salts, solvates,
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
salts, stereoisomers, prodrugs derivatives, or nitroderivatives of the
compounds listed above. In sorne cases, such as for example with
simvastatin, the active form of the statin is a metabolite formed in the body
of
a subject following administration. In other cases, statins are administered
in
their active form.
[0045] Phospholipases A2 (PLA2s) are a family of enzymes that catalyze
hydrolysis of the sn-2 fatty acyl ester bond of phospholipids to produce free
fatty acids and lysophospholipids such as arachidonic acid (AA) and
lysophosphatidylcholine. AA can then be converted into eicosanoids such as
prostagiandins, leukotrienes, thromboxanes, and lipoxins, while
lysophosphatidylcholine can be metabolized to lysophosphatidic acid (LPA) or
platelet-activating factor (PAF). PLA2s have been classified into several
groups based on factors such as cellular localization, amino acid sequence,
molecular mass, and calcium requirement for enzymatic activity (Ramoner
2005).
[0046] Secretory phospholipase A2 (sPLA2) is an extracellular or secreted
subgroup of PLA2 that plays a role in inducing inflammation. Eievated levels
of sPLA2 types IIA, IID, IIE, IIF, Ili, V, and X have been observed in all
stages
of atherosclerosis development and have been implicated in atherogenesis
based on their ability to degrade phospholipid (Kimura-Matsumoto 2007).
sPLA2 type IIA has been found to be expressed at vascular smooth muscle
cells and foam cells in human arteriosclerosis lesions, and this expression
has
been recognized to have a correlation with the development of arteriosclerosis
(Menschikowski 1995; Elinder 1997). Transgenic mice that express high
levels of human type 11A sPLA2 have increased LDL levels, decreased HDL
36
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
levels, and arteriosclerotic lesions (Ivandic 1999; Tietge 2000), and develop
arteriosclerosis at a higher rate compared to normal mice when given a high
fat diet (Ivandic 1999). Treatment with sPLA2 modifies LDL lipoproteins such
that they have higher affinity for extracellular matrix proteins (Camejo 1998;
Sartipy 1999; Hakala 2001), resulting in an increased retention of LDL
particles in the arterial wall. In addition, there is some evidence that sPLA2
remodels HDL, resulting in HDL catabolism (Pruzanski 1998). Increased
expression of sPLA2 type V has been shown to increase arteriosclerosis in
mice, while a deficiency of sPLA2 type V has been shown to reduce
arteriosclerosis (Rosengren 2006; Bostrom 2007).
[0047] sPLA2 expression has also been correlated with an increased risk of
development of CAD. Higher circulating levels of sPLA2, and of sPLA2 type
IIA specifically, have been observed in patients with documented CAD than in
control patients (Kugiyama 1999; Liu 2003; Boekholdt 2005; Chait 2005;
Hartford 2006). In addition, higher circulating levels of sPLA2 were found to
provide an accurate prognostic indicator for development of CAD in healthy
individuals (Mallat 2007). Measurement of sPLA2 activity has been shown to
be an independent predictor of death and new or recurrent myocardial
infarction in subjects with acute coronary syndrome, and provides greater
prognostic accuracy than measuring type IIA concentration only (Mallat 2005).
It has also been proposed that sPLA2 may have detrimental effects in the
setting of ischemic events. This is based largely on the finding of sPLA2
depositions in the necrotic center of infarcted human myocardium (Nijmeijer
2002).
37
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
[0048] As disclosed herein, administration of the sPLA2 inhibitor A-002,
which is a prodrug form of A-001, decreased plasma total cholesterol levels,
increased plasma HDL levels, and decreased atherosclerotic plaque
formation and aortic aneurysm in mice. In humans with CVD, twice daily
administration of A-002 decreased serum LDL, LDL particle, small LDL
particle, total cholesterol, and TG levels. Decreases in LDL were observed in
a diabetic subpopulation, as well as in a subpopulation exhibiting elevated
baseline LDL levels, while decreases in TG levels were observed in a
metabolic syndrome subpopulation. Additionally, administration of A-002 was
found to decrease serum levels of the inflammatory markers sPLA2, CRP, and
IL-6. Surprisingly, similar results were obtained with once a day
administration of A-002, which caused a decrease in LDL, non-HDL
cholesterol, total cholesterol, small LDL particle, oxidized LDL,
triglyceride,
and ApoB levels and an increase in LDL particle size. In addition, once a day
dosing with A-002 prevented the large increase in CRP levels observed in the
placebo group. Based on the experimental results disclosed herein showing
that administration of sPLA2 inhibitors decreases total cholesterol levels,
atherosclerotic plaque formation, and aortic aneurysm in mice, decreases
total cholesterol, non-HDL cholesterol, LDL, LDL particle, small LDL particle,
oxidized LDL particle, TG, and ApoB levels in humans, and increases LDL
particle size in humans, administration of a therapeutically effective amount
of
one or more sPLA2 inhibitors may be used to treat CVD and conditions
associated with CVD, such as for example dyslipidemia, atherosclerosis,
metabolic syndrome, CAD, CHD, and conditions associated with CAD and
CHD.
38
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
[0049] Provided herein in certain embodiments are methods of treating CVD
and conditions associated with CVD in a subject in need thereof by
administering a therapeutically effective amount of one or more sPLA2
inhibitors. In certain of these embodiments, administration of one or more
sPLA2 inhibitors may result in, among other effects, a decrease in cholesterol
levels, a decrease in TG levels, an increase in LDL particle size, and/or an
improvement in HDL/LDL ratio. Therefore, also provided herein are methods
of treating dyslipidemia in a subject in need thereof. In certain embodiments,
one or more sPLA2 inhibitors for use in these methods comprise A-001 or a
pharmaceutically acceptable prodrug, salt, polymorph, co-crystal, or solvate
of
A-001. In certain of these embodiments, the pharmaceutically acceptable
prodrug is A-002. In certain embodiments, administration of one or more
sPLA2 inhibitors may result in a decrease in one or more markers associated
with inflammation. Such markers include, but are not limited to, sPLA2, CRP,
and IL-6. Also provided herein are compositions comprising one or more
sPLA2 inhibitors for use in the methods disclosed herein, such as for example
in the treatment of CVD, conditions associated with CVD, and/or dyslipidemia
or to lower total cholesterol, LDL, non-HDL cholesterol, LDL particle, small
LDL particle, oxidized LDL, ApoB, and/or triglyceride levels or to raise HDL
levels. In certain embodiments, these compositions further comprise one or
more pharmaceutically acceptable carriers.
[0050] Compositions comprising one or more sPLA2 inhibitors for use in the
methods disclosed herein may be administered to a subject on a one-time
basis or in multiple administrations. In those embodiments wherein these
compositions are given in multiple administrations, the compositions may be
39
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
administered at set intervals over a particular time period determined in
advance, or they may be administered indefinitely or until a particular
therapeutic benchmark is reached, such as for example until a subject
exhibits cholesterol levels below a specified threshold. In certain
embodiments, the one or more sPLA2 inhibitors may be administered from
one or more times per day to once every week, once every month, or once
every several months. In certain embodiments, the one or more sPLA2
inhibitors may be administered twice a day, and in other embodiments the one
or more sPLA2 inhibitors may be administered once a day. As disclosed
herein, both twice a day and once a day administration of A-002 resulted in
significant decreases in serum lipid levels.
[0051] In certain embodiments, kits are provided that comprise one or more
sPLA2 inhibitors. In certain embodiments, the one or more sPLA2 inhibitors
comprise A-001 or a pharmaceutically acceptable prodrug, salt, polymorph,
co-crystal, or solvate of A-001, and in certain of these embodiments, the
pharmaceutically acceptable prodrug is A-002. In certain embodiments, the
kit provides instructions for usage, such as dosage or administration
instructions.
[0052] As disclosed herein, twice daily administration of A-002 was shown to
further decrease total cholesterol levels in mice and LDL, LDL particle, and
small LDL particle levels in humans that were receiving statin treatment.
Likewise, A-002 was shown to further decrease serum LDL levels in humans
that were receiving ezetimibe treatment. Similar results were obtained when
A-002 was administered once a day in combination with statins. These
results indicate that administration of an sPLA2 inhibitor unexpectedly causes
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
an additional decrease in cholesterol levels in subjects that are already
being
treated with another compound used in the treatment of CVD, such as a
statin. As further disclosed herein, administration of A-002 in conjunction
with
statins resulted in an unexpected synergistic decrease in total cholesterol
levels and aortic lesion formation in mice and in LDL and small LDL particle
levels in humans. The synergistic decrease in LDL following co-
administration of A-002 and statins was observed in the statin subpopulation
as a whole. In addition, synergism appeared to occur between A-002 and
each of the individual statins within the general statin subpopulation,
although
statistical analysis of this effect was complicated by the limited number of
test
and placebo subjects for each statin. These results indicate that the
synergistic decrease in LDL levels generated by co-administration of A-002
and statins is not limited to one particular statin, but rather occurs across
the
entire range of statins. Administration of A-002 in conjunction with ezetimibe
led to a similar synergistic decrease in LDL levels, indicating that the
synergism between A-002 and compounds used in the treatment of CVD is
not limited to statins. Therefore, methods are provided herein for treating
CVD and conditions associated with CVD, including dyslipidemia and
atherosclerosis, in a subject in need thereof by administering a
therapeutically
effective amount of one or more sPLA2 inhibitors and a therapeutically
effective amount of one or more compounds used in the treatment of CVD,
such as for example statins. Also provided herein are methods of increasing
the effectiveness of a compound used in the treatment of CVD, such as for
example a statin, by administering one or more sPLA2 inhibitors in conjunction
with the compound used in the treatment of CVD.
41
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
[0053] Provided herein in certain embodiments are methods of treating
dyslipidemia in a subject in need thereof by administering a therapeutically
effective aniount of one or more sPLA2 inhibitors and a therapeutically
effective amount of one or more compounds used in the treatment of CVD. In
certain embodiments, the one or more sPLA2 inhibitors comprise A-001 or a
pharmaceutically acceptable prodrug, salt, poiymorph, co-crystal, or solvate
of
A-001, and in certain of these embodiments the pharmaceutically acceptable
prodrug is A-002. In certain embodiments, the one or more compounds used
in the treatment of CVD comprise one or more statins or statin combination
drugs. In certain of these embodiments, the one or more statins are selected
from the group consisting of atorvastatin, simvastatin, rosuvastatin,
lovastatin,
pravastatin, cerivastatin, fluvastatin, mevastatin, and pitavastatin. and the
statin combination drugs are selected from the group consisting of
atorvastatin plus ezetimibe, atorvastatin plus amiodipine, atorvastatin plus
CP-529414, atorvastatin plus APA-01, simvastatin plus ezetimibe, simvastatin
plus extended release niacin, simvastatin plus MK-0524A, lovastatin plus
extended release niacin, rosuvastatin plus fenofibrate, pravastatin plus
fenofibrate, and statin plus TAK-457. In certain embodiments, the one or
more compounds used in the treatment of CVD comprise one or more non-
statin compounds selected from the group consisting of bile acid
sequestrants, fibrates, niacin or niacin derivatives, cholesterol absorption
inhibitors, cholesteryl ester transfer protein (CETP) inhibitors, microsomal
triglyceride transfer protein (MTP) inhibitors, squalene synthase inhibitors,
ACE inhibitors, angiotensin II receptor antagonists, beta-adrenergic blockers,
calcium channel blockers, and antithrombotics. In certain of these
42
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
embodiments, the one or more compounds used in the treatment of CVD
comprise ezetimibe. In certain embodiments, administration of one or more
sPLA2 inhibitors and one or more statins results in a decrease in cholesterol
levels, and in certain of these embodiments the decrease in cholesterol levels
is greater than that observed following administration of one or more sPLA2
inhibitors or one or more statins alone. In certain of these embodiments, the
decrease in cholesterol levels is synergistic, meaning that the decrease is
greater than the expected additive effect of one or more sPLA2 inhibitors and
one or more statins. In certain of these embodiments, administration of one or
more sPLA2 inhibitors and one or more statins results in an improved
HDL/LDL ratio, either by decreasing LDL levels, increasing HDL levels, or
both.
[0054] Provided herein in certain embodiments are methods of treating CVD
or conditions associated with CVD in a subject in need thereof by
administering a therapeutically effective amount of one or more sPLA2
inhibitors and a therapeutically effective amount of one or more compounds
used in the treatment of CVD. In certain embodiments, the one or more
sPLA2 inhibitors comprise A-001 or a pharmaceutically acceptable prodrug,
salt, polymorph, co-crystal, or solvate of A-001, and in certain of these
embodiments, the pharmaceutically acceptable prodrug is A-002_ In certain
embodiments, the one or more compounds used in the treatment of CVD
comprise one or more statins or statin combination drugs. In certain of these
embodiments, the one or more statins are selected from the group consisting
of atorvastatin, simvastatin, rosuvastatin, lovastatin, pravastatin,
cerivastatin,
fluvastatin, mevastatin, and pitavastatin, and the statin combination drugs
are
43
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
selected from the group consisting of atorvastatin plus ezetimibe,
atorvastatin
plus amlodipine, atorvastatin plus CP-529414, atorvastatin plus APA-01,
simvastatin plus ezetimibe, simvastatin plus extended release niacin,
simvastatin plus MK-0524A, lovastatin plus extended release niacin,
rosuvastatin plus fenofibrate, pravastatin plus fenofibrate, and statin plus
TAK-457. In certain embodiments, the one or more compounds used in the
treatment of CVD comprise one or more non-statin compounds selected from
the group consisting of bile acid sequestrants, fibrates, niacin or niacin
derivatives, cholesterol absorption inhibitors, cholesteiyi ester transfer
protein
(CETP) inhibitors, microsomal triglyceride transfer protein (MTP) inhibitors,
squalene synthase inhibitors. ACE inhibitors, angiotensin II receptor
antagonists, beta-adrenergic blockers, calcium channel blockers, and
antithrombotics. In certain of these embodiments, the one or more
compounds used in the treatment of CVD comprise ezetimibe. In certain
embodiments, treatment of CVD or a condition associated with CVD may be
associated with a decrease in cholesterol levels. In other embodiments,
however, treatment of certain forms of CVD may not be associated with a
concomitant decrease in cholesterol levels. For example, sPLA2 inhibitor and
statin administration may result in a decrease in atherosclerotic plaque
formation without a measurable decrease in cholesterol levels via a
mechanism such as inhibition of inflammation. In these embodiments,
treatment of CVD may be associated with a decrease in levels of one or more
inflammatory markers, such as for example sPLA2, CRP, and/or IL-6. In
certain embodiments, administration of a combination of one or more sPLA2
inhibitors and one or more statins results in a greater degree of treatment
than
44
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
that observed following administration of one or more sPLA2 inhibitors or
statins alone. In certain of these embodiments, the degree of treatment is
synergistic, meaning that the degree of treatment is greater than the expected
additive effect of one or more sPLA2 inhibitors and one or more statins.
[0055] Provided herein in certain embodiments are methods for increasing
the effectiveness of a compound used in the treatment of CVD by
administering one or more sPLA2 inhibitors in conjunction with the compound
used in the treatment of CVD. In certain embodiments, the one or more
sPLA2 inhibitors comprise A-001 or a pharmaceutically acceptable prodrug,
salt, polymorph, co-crystal, or solvate of A-001, and in certain of these
embodiments, the pharmaceutically acceptable prodrug is A-002. In certain
embodiments, the one or more compounds used in the treatment of CVD
comprise one or more statins or statin combination drugs. In certain of these
embodiments, the one or more statins are selected from the group consisting
of atorvastatin, simvastatin, rosuvastatin, lovastatin, pravastatin,
cerivastatin,
fluvastatin, mevastatin, and pitavastatin, and the statin combination drugs
are
selected from the group consisting of atorvastatin plus ezetimibe,
atorvastatin
plus amlodipine, atorvastatin plus CP-529414, atorvastatin plus APA-01,
simvastatin plus ezetimibe, simvastatin plus extended release niacin,
simvastatin plus MK-0524A, lovastatin plus extended release niacin,
rosuvastatin plus fenofibrate, pravastatin plus fenofibrate, and statin plus
TAK-457. In certain embodiments, the one or more compounds used in the
treatment of CVD comprise one or more non-statin compounds selected from
the group consisting of bile acid sequestrants, fibrates, niacin or niacin
derivatives, cholesterol absorption inhibitors, cholesteryl ester transfer
protein
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
(CETP) inhibitors, microsomal triglyceride transfer protein (MTP) inhibitors,
squalene synthase inhibitors, ACE inhibitors, angiotensin II receptor
antagonists, beta-adrenergic blockers, calcium channel blockers, and
antithrombotics. In certain of these embodiments, the one or more
compounds used in the treatment of CVD comprise ezetimibe. An increase in
effectiveness of a compound used in the treatment of CVD as used herein
refers to an increase the therapeutic effect of the compound, a decrease in
the dosage of the compound required to obtain a particular level of
therapeutic effect, or some combination thereof. For example, an increase in
effectiveness of a statin as used herein may refer to a greater decrease in
LDL levels following administration of a particular dosage of a statin, a
decrease in the dosage of statin required to bring about a particular decrease
in LDL levels, or some combination thereof.
[0056] Likewise, in certain embodiments methods are provided for increasing
the effectiveness of an sPLA2 inhibitor by administering one or more
compounds used in the treatment of CVD. In certain embodiments, the one
or more sPLA2 inhibitors comprise A-001 or a pharmaceutically acceptable
prodrug, salt, polymorph, co-crystal, or solvate of A-001, and in certain of
these embodiments, the pharmaceutically acceptable prodrug is A-002. In
certain embodiments, the one or more compounds used in the treatment of
CVD comprise one or more statins or statin combination drugs. In certain of
these embodiments, the one or more statins are selected from the group
consisting of atorvastatin, simvastatin, rosuvastatin, lovastatin,
pravastatin,
cerivastatin, fluvastatin, mevastatin, and pitavastatin, and the statin
combination drugs are selected from the group consisting of atorvastatin plus
46
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
ezetimibe, atorvastatin plus amlodipine, atorvastatin plus CP-529414,
atorvastatin plus APA-01, simvastatin plus ezetimibe, simvastatin plus
extended release niacin, simvastatin plus MK-0524A, lovastatin plus extended
release niacin, rosuvastatin plus fenofibrate, pravastatin plus fenofibrate,
and
statin plus TAK-457. In certain embodiments, the one or more compounds
used in the treatment of CVD comprise one or more non-statin compounds
selected from the group consisting of bile acid sequestrants, fibrates, niacin
or
niacin derivatives, cholesterol absorption inhibitors, cholesteryl ester
transfer
protein (CETP) inhibitors, microsomal triglyceride transfer protein (MTP)
inhibitors, squalene synthase inhibitors, ACE inhibitors, angiotensin !I
receptor
antagonists, beta-adrenergic blockers, calcium channel blockers, and
antithrombotics. In certain of these embodiments, the one or more
compounds used in the treatment of CVD comprise ezetimibe.
[0057] In certain embodiments, one or more sPLA2 inhibitors and one or
more compounds used in the treatment of CVD, such as for example one or
more statins, may be administered to a subject separately, i.e., as separate
compositions. In these embodiments, the one or more sPLA2 inhibitors and
the one or more compounds used in the treatment of CVD may be
administered simultaneously or sequentially. Further, the one or more sPLA2
inhibitors and the one or more compounds used in the treatment of CVD may
be administered at different times, and one compound may be administered
more frequently than another. In certain embodiments wherein the one or
more compounds used in the treatment of CVD are statins and wherein the
sPLA2 inhibitors and/or statins are given in multiple adniinistrations, one or
both may be administered anywhere from once or more times per day to once
47
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
every week, once every month, or once every several months. In certain
preferred embodiments, the one or rnore sPLA2 inhibitors and/or statins may
be administered twice a day, and in other preferred embodiments the one or
more sPLA2 inhibitors and/or statins may be administered once a day.
[0058] In certain embodiments, kits are provided that comprise one or more
sPLA2 inhibitors and one or more compounds used in the treatment of CVD.
In certain embodiments, the one or more sPLA2 inhibitors comprise A-001 or a
pharmaceutically acceptable prodrug, salt, polymorph, co-crystal, or solvate
of
A-001, and in certain of these embodiments, the pharmaceutically acceptable
prodrug is A-002. In certain embodiments, the one or more compounds used
in the treatment of CVD comprise one or more statins or statin combination
drugs. In certain of these embodiments, the one or more statins are selected
from the group consisting of atorvastatin, simvastatin, rosuvastatin,
lovastatin,
pravastatin, cerivastatin, fluvastatin, mevastatin, and pitavastatin, and the
statin combination drugs are selected from the group consisting of
atorvastatin plus ezetimibe, atorvastatin plus amiodipine, atorvastatin plus
CP-529414, atorvastatin plus APA-01, simvastatin plus ezetimibe, simvastatin
plus extended release niacin, simvastatin plus MK-0524A, lovastatin plus
extended release niacin, rosuvastatin plus fenofibrate, pravastatin plus
fenofibrate, and statin plus TAK-457. In certain embodiments, the one or
more compounds used in the treatment of CVD comprise one or more non-
statin compounds selected from the group consisting of bile acid
sequestrants, fibrates, niacin or niacin derivatives, cholesterol absorption
inhibitors, cholesteryl ester transfer protein (CETP) inhibitors, microsomal
triglyceride transfer protein (MTP) inhibitors, squalene synthase inhibitors,
48
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
ACE inhibitors, angiotensin II receptor antagonists, beta-adrenergic blockers,
calcium channel blockers, and antithrombotics. Within the kit, the one or
more sPLA2 inhibitors and one or more compounds used in the treatment of
CVD may be divided into separate compartments. For example, the kit may
comprise multiple bottles or packets, wherein each bottle or packet contains
either one or more sPLA2 inhibitors or one or more compounds used in the
treatment of CVD. In other embodiments, the one or more sPLA2 inhibitors
and one or more compounds used in the treatment of CVD may be found in a
single, undivided container. In certain embodiments, the kit provides
instructions for usage, such as dosage or administration instructions.
[0059] Provided herein in certain embodiments are pharmaceutical
compositions comprising a therapeutically effective amount of one or more
sPLA2 inhibitors and a therapeutically effective amount of one or more
compounds used in the treatment of CVD. In certain embodiments, the one
or more sPLA2 inhibitors comprise A-001 or a pharmaceutically acceptable
prodrug, salt, polymorph, co-crystal, or solvate of A-001, and in certain of
these embodiments, the pharmaceutically acceptable prodrug is A-002. In
certain embodiments, the one or more compounds used in the treatment of
CVD comprise one or more statins or statin combination drugs. In certain of
these embodiments, the one or more statins are selected from the group
consisting of atorvastatin, simvastatin, rosuvastatin, lovastatin,
pravastatin,
cerivastatin, fluvastatin, mevastatin, and pitavastatin, and the statin
combination drugs are selected from the group consisting of atorvastatin plus
ezetimibe, atorvastatin plus amlodipine, atorvastatin plus CP-529414,
atorvastatin plus APA-01, simvastatin plus ezetimibe, simvastatin plus
49
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
extended release niacin, simvastatin plus MK-0524A, lovastatin plus extended
release niacin, rosuvastatin plus fenofibrate, pravastatin plus fenofibrate,
and
statin plus TAK-457. In certain embodiments, the one or more compounds
used in the treatment of CVD comprise one or more non-statin compounds
selected from the group consisting of bile acid sequestrants, fibrates, niacin
or
niacin derivatives, cholesterol absorption iniiibitors, cholesteryl ester
transfer
protein (CETP) inhibitors, microsomal triglyceride transfer protein (MTP)
inhibitors, squalene synthase inhibitors, ACE inhibitors, angiotensin 11
receptor
antagonists, beta-adrenergic blockers, calcium channel blockers, and
antithrombotics. In certain of these embodiments, the one or more
compounds used in the treatment of CVD comprise ezetimibe.
[0060] Pharmaceutical compositions comprising one or more sPLA2
inhibitors and one or more compounds used in the treatment of CVD, such as
for example one or more statins, may be administered to a subject on a one-
time basis or in multiple administrations. In those ernbodirnents wherein the
compositions are given in multiple administrations, they may be administered
at set intervals over a particular time period determined in advance, or they
may be administered indefinitely or until a particular therapeutic benchmark
is
reached, such as for example until a subject exhibits cholesterol levels,
triglyceride levels, or inflammatory marker levels that are below a specified
threshold. In certain embodiments. the compositions may be administered
from once or more times per day to once every month or once every several
months. In certain of these embodiments, the compositions are administered
once or twice per day.
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
[0061] Pharmaceutical compositions comprising one or more sPLA2
inhibitors, one or more compounds used in the treatment of CVD, or one or
more sPLA2 inhibitors plus one or more compounds used in the treatment of
CVD as disclosed herein may be delivered to a subject by any administration
pathway known in the art, including but not limited to oral, aerosol, enteral,
nasal, ophthalmic, parenteral, or transdermal (e.g., topical cream or
ointment,
patch). "Parenteral" refers to a route of administration that is generally
associated with injection, including infraorbital, infusion, intraarterial,
intracapsular, intracardiac, intradermal, intramuscular, intraperitoneal,
intrapulmonary, intraspinal, intrasternal, intrathecal, intrauterine,
intravenous,
subarachnoid, subcapsular, subcutaneous, transmucosal, or transtracheal.
sPL.A2 inhibitor, statin, or combined sPLA2 inhibitor and statin
pharmaceutical
compositions as described herein may be administered in any
pharmaceutically acceptable form, including for example in the form of a
solid,
liquid solution, suspension, emulsion, dispersion, micelle, or liposome.
Preparations for injection may include sterile solutions ready for injection,
sterile dry soluble products, such as lyophilized powders, ready to be
combined with a solvent just prior to use, including hypodermic tablets,
sterile
suspensions ready for injection, sterile dry insoluble products ready to be
combined with a vehicle just prior to use, and sterile emulsions. The
solutions
may be either aqueous or nonaqueous. In certain embodiments, the
compositions may comprise one or more pharmaceutically acceptable carriers
or may be administered in conjunction with one or more pharmaceutically
acceptable carriers.
51
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
[0062] In certain embodiments, pharmaceutical compositions comprising one
or more sPLA2 inhibitors, one or more compounds used in the treatment of
CVD, or one or more sPLA2 inhibitors plus one or more compounds used in
the treatment of CVD as described herein may be formed into oral dosage
units, such as for example tablets, pills, or capsules. Such an oral dosage
unit niay comprise the active ingredients (e.g., A-002 and one or more
statins)
and one or more pharmaceutically acceptable carriers. As disclosed herein,
the feasibility of such an oral dosage unit was demonstrated by producing a
batch of tablets comprising A-002 and simvastatin as the active ingredients.
Each tablet was formulated with 250 mg of A-002 and 40 mg of simvastatin.
Pharmaceutically acceptable carriers utilized in formulating the tablets
included anhydrous lactose, lactose fast flo, and microcrystalline cellulose
as
diluents, hydroxypropyl cellulose as a binder, croscarmellose sodium as a
disintegrants, butylated hydroxyanisole as an antioxidant, magnesium
stearate as a lubricant, and polysorbate 80 as a surfactant. Water was used
as a solvent when forrnulating the tablet. The final tablets contained high
concentrations of both active ingredients as determined by HPLC analysis,
indicating that an sPLA2 inhibitor and one or more compounds used in the
treatment of CVD, such as for example one or more statins, may be
formulated into a single oral dosage unit. The specific tablet formulation
disclosed herein is provided as an example only. One of ordinary skill in the
art will recognize that the therapeutically acceptable carriers utilized in
the
formulation may be varied, and that such variations are routine in the art.
Likewise, the active ingredients may vary. As disclosed herein, administration
of A-002 in combination with a variety of statins causes a synergistic
decrease
52
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
in serum lipid levels. Therefore, in certain embodiments, A-002 may be
combined with any statin known in the art, including but not limited to
atorvastatin or atorvastatin calcium, cerivastatin, fluvastatin, lovastatin,
mevastatin, pitavastatin, pravastatin, rosuvastatin, and/or simvastatin. In
certain embodiments, an oral dosage unit may comprise a coating that
surrounds the active ingredients and pharmaceutically acceptable carrier(s).
[0063] In certain embodiments, pharmaceutical compositions comprising one
or more sPLA2 inhibitors, one or more compounds used in the treatment of
CVD, or one or more sPLA2 inhibitors plus one or more compounds used in
the treatment of CVD as disclosed herein may be administered via a time
release delivery vehicle, such as for example a time release oral dosage unit.
A"time release vehicle" as used herein refers to any delivery vehicle that
releases active agent (e.g., A-002 and one or more statins) at some time after
administration or over a period of time following administration rather than
immediately upon administration. Time release may be obtained by a coating
on the vehicle that dissolves over a set timeframe following administration.
In
certain embodiments, the time release vehicle may comprise multiple layers
of coating alternated with multiple layers of active ingredients, such that
each
layer of coating releases a certain volume of active ingredients as it
dissolves.
In other embodiments, sPLA2 inhibitor, statin, or combined sPLA2 inhibitor and
statin pharmaceutical compositions may be administered via an immediate
release delivery vehicle.
[0064] A therapeutically effective amount of an sPLA2 inhibitor or a
compound used in the treatment of CVD for use in the methods or
compositions disclosed herein may be determined for each compound
53
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
individually. For example, statins or statin combination drugs may be
administered or included in a pharmaceutical composition at a dosage that is
well known in the art to decrease cholesterol levels. One of skill in the art
will
recognize that in those ernbodiments wherein one or more compounds used
in the treatment of CVD are combined with one or more sPLA2 inhibitors in a
single composition, the amount of the compound used in the treatment of
CVD that constitutes a therapeutically effective amount may be different than
the amount of the compound that constitutes a therapeutically effective
amount of the compound when administered alone due to, for example,
interactions between the compound and the one or more sPLA2 inhibitors.
For example, the effective dosage of a statin for use in combination therapy
may be lower than the standard dosage for the statin when administered
alone. In this situation, one of skill in the art will be able to readily
determine a
therapeutically effective amount for the combination composition using
methods well known in the art. In certain embodiments, a therapeutically
effective amount of an sPLA2 inhibitor for use alone or in combination with
one
or more compounds used in the treatment of CVD may be from about 5 to
about 10,000 mg/dose. In certain of these embodiments, a therapeutically
effective amount of an sPLA2 inhibitor may be from about 25 to about 5,000
mg/dose, and in certain of these embodiments a therapeutically effective
amount may be from about 50 to about 500 mg/dose.
[0065] In certain embodiments, an sPLA2 inhibitor for use in the
compositions and methods disclosed herein may be an indole-based sPLA2
inhibitor, meaning that the compound contains an indole nucleus having the
structure:
54
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
N
H
[0066] A variety of indole-based sPLA2 inhibitors are known in the art. For
example, indole-based sPLA2 inhibitors that may be used in conjunction with
the present invention include but are not limited to those set forth in U.S.
Patent Nos. 5,654,326 (Bach); 5,733,923 (Bach); 5,919,810 (Bach);
5,919,943 (Bach); 6,175,021 (Bach); 6,177,440 (Bach); 6,274,578 (Denney);
and 6,433,001 (Bach), the entire disclosures of which are incorporated by
reference herein. Methods of making indole-based sPLA2 inhibitors are set
forth in, for example, U.S. Patent Nos. 5,986,106 (Khau); 6,265,591
(Anderson); and 6,380,397 (Anderson), the entire disclosures of which are
incorporated by reference herein. sPLA2 inhibitors for use in the present
invention may be generated using these synthesis methods, or using any
other synthesis method known in the art. In certain embodiments, sPLA2
inhibitors for use in the present invention may be sPLA2 type IIA, type V,
and/or type X inhibitors. Various examples of indole-based sPLA2 inhibitors
are set forth below. These examples are merely provided as illustrations of
the types of inhibitors that may be used in conjunction with the present
invention, and as such are not meant to be limiting. One of ordinary skill in
the art will recognize that a variety of other indole-based sPLA2 inhibitors
may
be used.
[0067] In certain embodiments, sPLA2 inhibitors for use in the current
invention are 1 H-indole-3-glyoxylamide compounds having the structure:
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
X
R4 X
Rs
NH2
.----~"
R6
RZ
N
R7
Ri
wherein:
each X is independently oxygen or sulfur;
R, is selected from the group consisting of (a), (b), and (c), wherein:
(a) is C7-C20 alkyl, C7-C20 alkenyl, C7-C20 alkynyl, carbocyclic
radicals, or heterocyclic radicals;
(b) is a member of (a) substituted with one or more independently
selected non-interfering substituents; and
(c) is the group -(L) -R80, where, -(L)- is a divalent linking group of
1 to 12 atoms selected from carbon, hydrogen, oxygen, nitrogen, and sulfur,
wherein the combination of atoms in -(L)- are selected from the group
consisting of (i) carbon and hydrogen only, (ii) sulfur only, (iii) oxygen
only, (iv)
nitrogen and hydrogen only, (v) carbon, hydrogen, and sulfur only, and (vi)
carbon, hydrogen, and oxygen only; and where R80 is a group selected from
(a) or (b);
R2 is hydrogen, halo, C,-C3 alkyl, C3-C4 cycloalkyl, C3-C4 cycloalkenyl, -O-
(C1-C2 alkyl), -S-(C1-C2 alkyl), or a non-interfering substituent having a
total
of 1 to 3 atoms other than hydrogen;
R4 and R5 are independently selected from the group consisting of hydrogen,
a non-interfering substituent, and -(La)-(acidic group), wherein -(L,)- is an
56
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
acid linker having an acid linker length of 1 to 4; provided that at least one
of
R4 and R5 must be -(L,)-(acidic group);
R6 and R7 are each independently selected from hydrogen, non-interfering
substituents, carbocyclic radicals, carbocyclic radicals substituted with non-
interfering substituents, heterocyclic radicals, and heterocyclic radicals
substituted with non-interfering substituents;
provided that for any of the groups R,, R6, and R7, the carbocyclic radical is
selected from the group consisting of cycloalkyl, cycloalkenyl, phenyl,
naphthyl, norbornanyl, bicycloheptadienyl, tolulyl, xylenyl, indenyl,
stilbenyl,
terphenylyl, diphenylethylenyl, phenyl-cyclohexenly, acenaphthylenyl, and
anthracenyl, biphenyl, bibenzylyl and related bibenzylyl homologues
represented by the formula (bb),
0_(CH2)n__O(bb)
where n is a number from 1 to 8; provided, that for any of the groups Ri, R6,
and R7, the heterocyclic radical is selected from the group consisting of
pyrrolyi, furanyl, thiophenyl, pyrazolyl, imidazolyl, phenylimidazolyl,
triazolyl,
isoxazolyl, oxazolyl, thiazolyl, thiadiazolyl, indolyl, carbazolyl,
norharmanyl,
azaindolyl, benzofuranyl, dibenzofuranyl, thianaphtheneyl, dibenzothiophenyl,
indazolyl, imidazo(1.2-A)pyridinyl, benzotriazolyl, anthranilyl, 1,2-
benzisoxazolyl, benzoxazolyl, benzotriazolyl, purinyl, pryidinyl, dipyridylyl.
phenylpyridinyl, benzylpyridinyl, pyrimidinyl, phenylpyrimidinyl, pyrazinyl,
1,3,5-triazinyl, quinolinyl, phthalazinyl, quinazolinyl, and quinoxalinyl; and
provided that for the groups Rl, R2, R4, R,
, R6, and R7 the non-interfering
substituent is selected from the group consisting of C1-C6 alkyl, C2-C6
57
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
alkenyl, C2-C6 alkynyl, C7-C12 aralkyl, C7-C1Z alkaryl, C3-CII cycloalkyl, C3-
CB
cycloalkenyl, phenyl, tolulyl, xylenyl, biphenyl, Cl-C6 alkoxy, C2-CG
alkenyloxy, C2-C6 alkynyloxy, C2-C12 alkoxyalkyl, C2-C12 alkoxyalkyloxy, C2-
C12 alkylcarbonyl, C2-C12 alkylcarbonylamino, C2-C12 alkoxyamino, C2-C12
alkoxyaminocarbonyl, C2-C12 alkylamino, C,-C6 alkylthio, C2-C12
alkylthiocarbonyl, C1-C6 alkylsulfinyl, C1-C6 alkylsulfonyl, C2-C6 haloalkoxy,
CI-C6 haloalkylsulfonyl, C2-C6 haloalkyl, C1-C6 hydroxyalkyl, -C(O)O(C1-C6
alkyl), -(CH2)n O-(C,-C6 alkyl), benzyloxy, phenoxy, phenylthio, -
(CONHSO2R), -CHO, amino, amidino, bromo, carbamyl, carboxyl,
carbalkoxy, -(CH2)õ -COzH, chloro, cyano, cyanoguanidinyl, fluoro,
guanidino, hydrazide, hydrazino, hydrazido, hydroxy, hydroxyamino, iodo,
nitro, phosphono, -SO3H, thioacetal, thiocarbonyl, and C1-Cr6 carbonyl, where
n is from 1 to 8;
and pharmaceutically acceptable salts, solvates, prodrug derivatives,
racemates, tautomers, or optical isomers thereof.
[0068] In certain of these embodiments, -(L)- has the formula:
Ra~
Z c
I
R82 p
wherein R81 and R82 are each independently selected from the group
consisting of hydrogen, C1-C,o alkyl, carboxy, carbalkoxy, and halo; p is a
number from 1 to 5; and Z is selected from the group consisting of a bond, -
(CHZ)-, -0-, -N(C,-C,o alkyl)-, -NH-, and -S-.
[0069] In certain of these embodiments wherein R4 is -(La)-(acidic group),
the acid linker -(La)- has the formula:
58
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
R83
I
Q -_-_" C
(
R84
wherein 0 is selected from the group consisting of -(CH2)-, -0-, -NH-, and
-S-; and R83 and R84 are each independently selected from the group
consisting of hydrogen, C,-C,o alkyl, aryl, Cl-C1o alkaryl, C,-C,o aralkyl,
hydroxy, and halo.
[0070] In certain of these embodiments wherein R5 is -(La)-(acidic group),
the acid linker -(La)- has the formula:
R 85
Q-~C~phenylene S
I
R86
wherein r is a number from 2 to 7; s is 0 or 1; Q is selected from the group
consisting of -(CH2)-, -0-, -NH-, and -S-; and R85 and R86 are each
independently selected from the group consisting of hydrogen, Cl-C10 alkyl,
aryl, Cl-C1o alkaryl, C,-C,o aralkyl, carboxy, carbalkoxy, and halo.
[0071] In certain embodiments, a 1 H-indole-3-giyoxylamide compound for
use in the present invention is selected from the group consisting of: ((3-(2-
Amino-l,2-dioxoethyl)-2-ethyl-l-(phenylmethyl)-1 H-indol-4-yl)oxy)acetic acid;
[[3-(2-Amino-1,2-dioxoethyl)-2-ethyl-1-(phenylmethyl)-1 H-indol-4-
yi]oxy]acetic
acid methyl ester; ((3-(2-Amino-l,2-dioxoethyl)-2-methyl-l-(phenylmethyl)-1H-
indol-4-yi)oxy)acetic acid; dl-2-((3-(2-Amino-1,2-dioxoethyl)-2-methyl-l-
(phenyimethyl)-1 H-indol-4-yi) oxy)propanoic acid; ((3-(2-Amino-1,2-
dioxoethyl)-1-((1,1'-biphenyl)-2-ylmethyl)-2-methyl-1 H-indol-4-yl)oxy)acetic
59
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
acid; ((3-(2-Amino-1,2-dioxoethyl)-1-((1,1'-biphenyl)-3-ylmethyl)-2-methyl-lH-
indol-4-yl)oxy)acetic acid; ((3-(2-Amino-1,2-dioxoethyl)-1-((1,1'-biphenyl)-4-
ylmethyl)-2-methyl-1 H-indol-4-yl)oxy)acetic acid; ((3-(2-Amino-1,2-
dioxoethyl)-
1-((2,6-dichlorophenyl)methyl)-2-methyl-1 H-indol-4-yi)oxy)acetic acid; ((3-(2-
Amino-l,2-dioxoethyl)-1-(4(-fluorophenyl)methyl)-2-methyl-1 H-indol- 4-
yI)oxy)acetic acid; ((3-(2-Amino-1,2-dioxoethyl)-2-methyl-l-((1-
naphthalenyl)methyl)-1 H-indol- 4-yl)oxy)acetic acid; ((3-(2-Amino-1,2-
dioxoethyl)-1-((3-chlorophenyl)methyl)-2-ethyl-1 H-indol-4-yl)oxy)acetic acid;
((3-(2-Amino-1,2-dioxoethyl)-1-((1,1'-biphenyl)-2-ylmethyl)-2-ethyl-1 H-indol-
4-
yI)oxy)acetic acid; ((3-(2-amino-l,2-dioxoethyl)-1-((1,1'-biphenyl)-2-
ylmethyl)-
2-propyl-1 H-indol-4-yl)oxy)acetic acid; ((3-(2-Amino-1,2-dioxoethyl)-2-
cyclopropyl-1-(phenylmethyl)-1H-indol-4-yl) oxy)acetic acid; ((3-(2-Amino-1,2-
dioxoethyl)-1-((1,1'-biphenyl)-2-ylmethyl)-2-cyclopropyl-1 H-indol-4-
yl)oxy)acetic acid; and 4-((3-(2-Amino-l,2-dioxoethyl)-2-ethyl-l-
(phenylmethyl)-1 H-indol-5-yl)oxy) butanoic acid, or pharmaceutically
acceptable salts, solvates, prodrug derivatives, racemates, tautomers, or
optical isomers thereof.
[0072] In certain embodiments, sPLA2 inhibitors for use in the current
invention are 1 H-indole-3-glyoxylamide compounds having the structure:
x
NH2
R, R6 ,
eN
7
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
wherein:
both X are oxygen;
R, is selected from the group consisting of:
Rlo)t
~CH2) 1-2
and
( H 2C~
CH?)Q 2 /
wherein Rlo is a radical independently selected from halo, C,-C,o alkoxy, -S-
( Cl-Clo alkyl), and Cl-C1o haloalkyl, and t is a nurnber from 0 to 5;
R2 is selected from the group consisting of halo, cyclopropyl, methyl, ethyl,
and propyl;
R4 and R5 are independently selected from the group consisting of hydrogen,
a non-interfering substituent, and -(La)-(acidic group), wherein -(La)- is an
acid linker; provided that the acid linker -(La)- for RG is selected from the
group consisting of:
~H2C-CH2[---
61
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
cl-i 3
Q
and
and
provided that the acid linker -(La)- for R5 is selected from the group
consisting
of:
R84
o
R 85
R84
R85
R84
H
N (
R85
62
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
R84
[cH22 C
i
R85
R 8a
o-C ( CH2)
1-3
R85
R84
I
S-C ( CH2)
1-3
R85
R84
H I
N C (CH2)
l 1-3
R85 and
R84
H2 I
C -C ( CHZ)1 3
R85
wherein R84 and R85 are each independently selected from the group
consisting of hydrogen, Cl-C1o alkyl, aryl, C,-C,o alkaryl, C,-C,a aralkyl,
carboxy, carbalkoxy, and halo; provided that at least one of Ra and R5 must
be -(La)-(acidic group), and (acidic group) on -(La)-(acidic group) of R4 or
R5
is selected from -COzH, -SO3H, or -P(O)(OH)2;
R6and R7 are each independently selected from the group consisting of
63
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
hydrogen and non-interfering substituents, with the non-interfering
substituents being selected from the group consisting of: C,-C6 alkyl, C2-C6
alkenyl, C2-C6 alkynyl, C7-C12 aralkyl, C7-C12 alkaryl, C3-Ca cycloalkyi, C3-
C8
cycloalkenyl, phenyl, tolulyl, xylenyl, biphenyl, Cl-C6 alkoxy, C2-C6
alkenyloxy, C2-C6 alkynyloxy, C2-C12 alkoxyalkyl, CZ-C,z alkoxyalkyloxy, C2-
C1Z alkylcarbonyl, C2-C12 alkylcarbonylamino, C2-C12 alkoxyamino, C2-C12
alkoxyaminocarbonyl, C2-C12 alkylamino, C1-C6 alkylthio, C2-C12
alkylthiocarbonyl, Cl-C6 alkylsulfinyl, C1-C6 alkylsulfonyl, C2-C6 haloalkoxy,
Cl-C6 haloalkylsulfonyl, CZ-C6 haloalkyl, C,-Ce, hydroxyalkyl, -C(O)O(C1-C6
alkyl), -(CHZ)n-O-(C1-Cs alkyl), benzyloxy, phenoxy, phenylthio, -
(CONHSO2R), -CHO, amino, amidino, bromo, carbamyl, carboxyl,
carbalkoxy, -(CH2) n CO2H, chloro, cyano, cyanoguanidinyl, fluoro, guanidino,
hydrazide, hydrazino, hydrazido, hydroxy, hydroxyamino, iodo, nitro,
phosphono, -SO3H, thioacetal, thiocarbonyl, and Cl-C6 carbonyl; wherein n is
from 1 to 8:
and pharmaceutically acceptable salts, solvates, prodrug derivatives,
racemates, tautomers, or optical isomers thereof.
[0073] In certain embodiments, 1 H-indole-3-glyoxylamide compounds for use
in the present invention are selected from the group consisting of: ((3-(2-
Amino-1,2-dioxoethyl)-2-methyl-1-(phenylmethyl)-1 H-indol-4-yl)oxy)acetic
acid; ((3-(2-Amino-1,2-dioxoethyl)-2-rnethyl-1-(phenylrnethyl)-1 H-indol-4-
yl)oxy)acetic acid methyl ester; d1-2-((3-(2-Amino-l,2-dioxoethyl)-2-methyl-1-
(phenylmethyl)-1 H-indol-4-yl) oxy)propanoic acid; dI-2-((3-(2-Arnino-1,2-
dioxoethyl)-2-methyi-1-(phenylmethyl)-1 H-indol-4-yi) oxy)propanoic acid
methyl ester; ((3-(2-Amino-1,2-dioxoethyl)-1-((1,1'-biphenyl)-2-ylmethyl)-2-
64
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
methyl-lH-in dol-4-yl)oxy)acetic acid; ((3-(2-Amino-1,2-dioxoethyl)-1-((1,1'-
biphenyl)-2-ylmethyl)-2-methyl-1 H-in dol-4-yl)oxy)acetic acid methyl ester;
((3-
(2-Amino-l,2-dioxoethyl)-1-((1,1'-biphenyl)-3-ylmethyl)-2-methyl-1 H-in dol-4-
yl)oxy)acetic acid; ((3-(2-Amino-1,2-dioxoethyl)-1-((1,1'-biphenyl)-3-
ylmethyl)-
2-methy!-1 H-in dol-4-yl)oxy)acetic acid methyl ester; ((3-(2-Amino-1,2-
dioxoethyl)-1-((1,1'-biphenyl)-4-ylmethyl)-2-methyl-1 H-in dol-4-yl)oxy)acetic
acid; ((3-(2-Amino-l,2-dioxoethyl)-1-((1,1'-biphenyl)-4-ylmethyl)-2-methyl-lH-
in dol-4-yl)oxy)acetic acid methyl ester; ((3-(2-Amino-l,2-dioxoethyl)-1-((2,6-
dichlorophenyl)methyl)-2-methyl-1 H-in dol-4-yl)oxy)acetic acid; ((3-(2-Amino-
1,2-dioxoethyl)-1-((2,6-dichlorophenyl)methyl)-2-methyl-1 H-in dol-4-
yl)oxy)acetic acid methyl ester; ((3-(2-Amino-l,2-dioxoethyl)-1-(4(-
fluorophenyl)methyl)-2-methyl-1 H-indol- 4-yl)oxy)acetic acid; ((3-(2-Amino-
1, 2-dioxoethyi )-1-(4(-fluorophenyl)methyl)-2-methyl-1 H-indol- 4-
yl)oxy)acetic
acid methyl ester; ((3-(2-Amino-1,2-dioxoethyl)-2-methyl-1-((1-
naphthalenyl)methyl)-1 H-indol- 4-yl)oxy)acetic acid; ((3-(2-Amino-1,2-
dioxoethyl)-2-methyl-1-((1-naphthalenyl)methyl)-1 H-indol- 4-yl)oxy)acetic
acid
methyl ester; ((3-(2-Amino-l,2-dioxoethyl)-1-((3-chlorophenyl)methyl)-2-ethyl-
1H-indol-4 -yl)oxy)acetic acid; ((3-(2-Amino-1,2-dioxoethyl)-1-((3-
chlorophenyl)methyl)-2-ethyl-1 H-indol-4 -yl)oxy)acetic acid methyl ester; ((3-
(2-Amino-l,2-dioxoethyl)-1-((1,1'-biphenyl)-2-ylmethyl)-2-ethyl-1 H-ind ol-4-
yl)oxy)acetic acid; ((3-(2-Amino-1,2-dioxoethyl)-1-((1,1'-biphenyl)-2-
ylmethyl)-
2-ethyl-1 H-ind ol-4-yl)oxy)acetic acid methyl ester; ((3-(2-amino-1,2-
dioxoethyl)-1-((1,1'-biphenyl)-2-ylmethyl)-2-propyl-1 H-in dol-4-yi)oxy)
acetic
acid; ((3-(2-amino-l,2-dioxoethyl)-1-((1,1'-biphenyl)-2-ylmethyl)-2-propyl-1H-
in
dol-4-yl)oxy)acetic acid methyl ester; ((3-(2-Amino-1,2-dioxoethyl)-2-
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
cyclopropyl-1-(phenylmethyl)-1H-indol-4-yl) oxy)acetic acid; ((3-(2-Amino-1,2-
dioxoethyl)-2-cyclopropyl-1-(phenylmethyl)-1 H-indol-4-yl) oxy)acetic acid
methyl ester; ((3-(2-Amino-1,2-dioxoethyl)-1-((1,1'-biphenyl)-2-ylmethyl)-2-
cyclopropyl- 1 H-indol-4-yi)oxy)acetic acid; ((3-(2-Amino-1,2-dioxoethyl)-1-
((1,1'-biphenyl)-2-ylmethyl)-2-cyclopropyl- 1 H-indol-4-yl)oxy)acetic acid
methyl
ester; 4-((3-(2-Amino-1,2-dioxoethyl)-2-ethyl-1-(phenylmethyl)-1 H-indol-5-
yl)oxy) butanoic acid; 4-((3-(2-Amino-1,2-dioxoethyl)-2-ethyl-l-(phenylmethyl)-
1 H-indol-5-yl)oxy) butanoic acid tert-butyl ester, or pharmaceutically
acceptable salts, solvates, prodrug derivatives, racemates, tautomers, or
optical isomers thereof.
[0074] In certain embodiments, sPLA2 inhibitors for use in the current
invention are 1 H-indole-3-glyoxylamide compounds having the structure:
x
x
R4 NH2
Rs
R
2
R
6 N
R7 R,
wherein:
each X is independently oxygen or sulfur;
R, is selected from groups (a), (b), and (c) wherein:
(a) is C7-C20 alkyl, C7-C20 alkenyl, C,-C?o alkynyl, carbocyclic
radical, or heterocyclic radical;
(b) is a member of (a) substituted with one or more independently
selected non-interfering substituents; and
66
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
(c) is the group -(L)-R80, wherein -(L)- is a divalent linking group of
1 to 12 atoms selected from carbon, hydrogen, oxygen, nitrogen, and sulfur;
wherein the combination of atoms in -(L)- are selected from the group
consisting of (i) carbon and hydrogen only, (ii) sulfur only, (iii) oxygen
only, (iv)
nitrogen and hydrogen only, (v) carbon, hydrogen, and sulfur only, and (vi)
and carbon, hydrogen, and oxygen only; and where R80 is a group selected
from (a) or (b);
R2 is selected from the group consisting of hydrogen, halo, C,-C3 alkyl, C3-C4
cycloalkyl, C3-C4 cycloalkenyl, -O-(C1-C2 alkyl), -S-(C1-C2 alkyl), and a
non-interfering substituent having a total of 1 to 3 atoms other than
hydrogen;
R4 and R5 are independently selected from the group consisting of hydrogen,
a non-interfering substituent, and the group -(La)-(acidic group), wherein -
(La)- is an acid linker having an acid linker length of 1 to 4; provided that
at
least one of R4 and R5 is -(La)-(acidic group);
R6 and R7 are each independently selected from the group consisting of
hydrogen, non-interfering substituents, carbocyclic radicals, carbocyclic
radicals substituted with non-interfering substituents, heterocyclic radicals,
and heterocyclic radicals substituted with non-interfering substituents;
and pharmaceutically acceptable salts, solvates, prodrug derivatives,
racemates, tautomers, or optical isomers thereof.
[0075] In certain embodiments, sPLA2 inhibitors for use in the current
invention are methyl ester prodrug derivatives of 1 H-indole-3-glyoxylamide
compounds having the structure:
67
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
X
NH2
aN
R5 R s R7 Ri
wherein:
both X are oxygen;
R, is selected from the group consisting of:
Rlo
~CH2)1-2
and
`H2C<2H C
H2)0-2 \ /
wherein Rlo is a radical independently selected from halo, C1-Clo alkyl, C1-
ClQ alkoxy,-S-( Cl-Clo alkyl), and C,-C,o haloalkyl, and t is a number from 0
to 5;
R2 is selected from the group consisting of halo, cyclopropyl, methyl, ethyl,
and propyl;
R4 and R5 are independently selected from the group consisting of hydrogen,
a non-interfering substituent, and -(La)-(acidic group), wherein -(La)- is an
acid linker; provided that the acid linker -(La)- for R4 is selected from the
group consisting of:
68
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
4O - CH 2 ~-
4S-CH2]--
4N-CH2-}-
f H?C-CH2[-
CH3
and
-~O and
provided that the acid linker -(La)- for R5 is selected from the group
consisting
of:
R84
O`j
R85
[R8a
S- R85
69
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
R84
H_I
N (
R85
R 84
CH2)2 C
I
R85
R84
O-C (CH2)
1-3
R 8s
R84
S-C (CH2) 1-3
R 8,
R8,
H I
N-C (CH2)
1 1-3
R85 and
R 84
H2 I
C -C (CH2)1 3
R85
wherein R84 and R85 are each independently selected from the group
consisting of hydrogen, C,-C,o alkyl, aryl, C1-Clo alkaryl, C1-Clo aralkyl,
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
carboxy, carbalkoxy, and halo; provided that at least one of R4 and R5 must
be -(La)-(acidic group), and (acidic group) on -(L,,)-(acidic group) of R4 or
R5
is selected from -COzH, -SO3H, or -P(O)(OH)2;
R6 and R7 are each independently selected from the group consisting of
hydrogen and non-interfering substituents, with the non-interfering
substituents being selected from the group consisting of: Cj-C6 alkyl, C2-C6
alkenyl, C2-C6 alkynyl, C7-C12 aralkyl, C7-C12 alkaryl, C3-C8 cycloalkyl, C3-
C8
cycloalkenyl, phenyl, tolulyi, xylenyl, biphenyl, C1-C6 alkoxy, C2-C6
alkenyloxy, C2-C6 alkynyloxy, C2-C12 alkoxyalkyl, C2-C12 alkoxyalkyloxy, CZ-
C1Z alkylcarbonyl, C2-C12 alkylcarbonylamino, C2-C12 alkoxyamino, C2-C12
alkoxyaminocarbonyl, C2-C12 alkylamino, C1-C6 alkylthio, C2-C12
alkylthiocarbonyl, Cl-C6 alkylsulfinyl, C1-C6 alkylsulfonyl, CZ-C6 haloalkoxy,
Cl-C6 haloalkylsulfonyl, C2-C6 haloalkyl, C1-C6 hydroxyalkyl, -C(O)O(Cj-C6
alkyl), -(CH2)n O-(C,-C6 alkyl), benzyloxy, phenoxy, phenylthio, -
(CONHSO2R), -CHO, amino, amidino, bromo, carbamyl, carboxyl,
carbalkoxy, -(CH2) n-CO2H, chloro, cyano, cyanoguanidinyl, fluoro, guanidino,
hydrazide, hydrazino, hydrazido, hydroxy, hydroxyamino, iodo, nitro,
phosphono, -SO3H, thioacetal, thiocarbonyl, and Ci-C6 carbonyl; wherein n is
from 1 to 8;
and pharmaceutically acceptable salts, solvates, prodrug derivatives,
racemates, tautomers, or optical isomers thereof.
[0076] In certain embodiments, sPLA2 inhibitors for use in the current
invention are (acyloxy) alkyl ester prodrug derivatives of 1 H-indole-3-
glyoxylamide compounds having the structure:
71
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
x
X
R4 NH2
R5
R2
R
6 N
R; Ri
wherein:
both X are oxygen;
R, is selected from the group consisting of:
R1o
--(CH2)1-2
and
(H2C~
CH2) 0-2
wherein Rlo is a radical independently selected from halo, Cl-C1o alkyl, Cl-
Clo alkoxy,-S-( C,-C,o alkyl), and Cl-C1o haloalkyl, and t is a number from 0
to 5;
R2 is selected from the group consisting of halo, cyclopropyl, methyl, ethyl,
and propyl;
R4 and R5 are independently selected from the group consisting of hydrogen,
a non-interfering substituent, and -(La)-(acidic group), wherein -(L,)- is an
acid linker; provided that the acid linker -(La)- for R4 is selected from the
group consisting of:
72
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
O CH?-}-
- 4 S-CH2~-
4 N CH2~-
-4 H2C -- CH?}---
CH3
and
and
provided that the acid linker -(La)- for R5 is selected from the group
consisting
of:
Ra4
O \ /
R85
[R8a
S Ra5
73
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
R84
HI
N_ R85
R8ic;
( [cH22 C I
R85
R84
O-C (CH2)
1-3
Rss
R84
s-C (CH2)
1-3
R85
R84
N C (CH2)
1-3
R85 and
R84
H2
C -C (CFiz)1 3
R85
wherein R84 and R8, are each independently selected from the group
consisting of hydrogen, Cl-C1o alkyl, aryl, Cl-Clo alkaryl, Cl-C1o aralkyl,
74
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
carboxy, carbalkoxy, and halo; provided that at least one of R4 and R5 must
be -(La)-(acidic group), and (acidic group) on -(La)-(acidic group) of R4 or
R5
is selected from -C02H, -SO3H, or -P(O)(OH)2;
R6 and R7 are each independently selected from the group consisting of
hydrogen and non-interfering substituents, with the non-interfering
substituents being selected from the group consisting of: C1-C6 alkyl, C2-C6
alkenyl, C2-C6 alkynyl, C7--C12 aralkyl, C7-C12 alkaryl, C3-C8 cycloalkyl, C3-
C8
cycloalkenyl, phenyl, tolulyl, xylenyl, biphenyl, C1-C6 alkoxy, C2-C6
alkenyloxy, C2-C6 alkynyloxy, C2-C12 alkoxyalkyl, C2-C12 alkoxyalkyloxy, C2-
C12 alkylcarbonyl, CZ-C,2 alkylcarbonylamino, C2-C12 alkoxyamino, CZ-C,z
alkoxyaminocarbonyl, C2-C12 alkylamino, Cl-C6 alkylthio, C2-C12
alkylthiocarbonyl, C1-C6 alkylsulfinyl, C1-C6 alkylsulfonyl, C2-C6 haloalkoxy,
Cl-C6 haloalkylsulfonyl, C2-C6 haloalkyl, C1-C6 hydroxyalkyl, -C(O)O(C1-C6
alkyl), -(CH2)n-O-(C1-C6 alkyl), benzyloxy, phenoxy, phenylthio, -
(CONHSO2R), -CHO, amino, amidino, bromo, carbamyl, carboxyl,
carbalkoxy, -(CH2) -CO2H, chloro, cyano, cyanoguanidinyl, fluoro, guanidino,
hydrazide, hydrazino, hydrazido, hydroxy, hydroxyamino, iodo, nitro,
phosphono, -SO3H, thioacetal, thiocarbonyl, and Cl-C6 carbonyl; wherein n is
from 1 to 8;
and pharmaceutically acceptable salts, solvates, prodrug derivatives,
racemates, tautomers, or optical isomers thereof.
[0077] In certain embodiments, sPLA2 inhibitors for use in the current
invention are substituted tricyclics having the structure:
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
CORi
R2
`5\ 4 3
6
A Z 2
7
R3 g N
I
CH?R4
wherein:
R3 is selected from the group consisting of-NHNH2 and -NH2;
R2 is selected from the group consisting of -OH and -O(CH2) n,R5; wherein R5
is selected from the group consisting of H, -CO2H. -C02(C1-C4 alkyl), -
SO3H, -S03(C1-C4 alkyl), tetrazolyl, -CN, -NH2, -NHSO2R15, -
CONHS02R15, phenyl, phenyl substituted with -CO2H or -C02(C1-C4)alkyl,
and
O
(I (R6R,)
wherein R6 and R7 are each independently selected from the group consisting
of -OH, -O(C,-Cq)alkyl; R15 is selected from the group consisting of -(Cl-
C6)alkyl and -CF3; and m is 1-3;
R3 is selected from the group consisting of H, -O(C,-Ca)alkyl, halo, -(Cl-
C6)alkyl, phenyl, -(C1-C4)alkylphenyl, phenyl substituted with -(C,-C6)alkyl,
halo, or -CF3, -CH2OSi(Cj-C6)alkyl, furyl, thiophenyl, -(C,-
C6)hydroxyalkyl, and -(CH2),R8; wherein R8 is selected from the group
consisting of H, -CONH2, -NRqR,O, -CN, and phenyl; wherein R9 and Rlo are
each independently -(C,-C4)alkyl or -phenyl(C,-C4)alkyl; and n is 1 to 8;
76
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
R4 is selected from the group consisting of H, -(C; C,4)alkyl, -(C3-
C14)cycloalkyl, pyridyl, phenyl, and phenyl substituted with -(C,-C6)alkyl,
halo,
-CF3, -OCF3, -(C1-C4)alkoxy, -CN, -(C,-C4)alkylthio, phenyl(C,-C4)alkyl, -
(Ca-C4)alkylphenyl, phenyl, phenoxy, or naphthyl;
A is selected from the group consisting of phenyl and pyridyl wherein the
nitrogen is at the 5-, 6-, 7-, or 8-position;
Z is selected from the group consisting of cyclohexenyl, phenyl, pyridyl
wherein the nitrogen is at the 1-, 2-, or 3-position, and a 6-membered
heterocyclic ring having one heteroatom selected from the group consisting of
sulfur and oxygen at the 1-, 2-, or 3-position and nitrogen at the 1-, 2-, 3-,
or
4- position, or wherein one carbon on the heterocyclic ring is optionally
substituted with =0; and wherein one of A or Z is a heterocyclic ring;
and pharmaceutically acceptable salts, solvates, prodrug derivatives,
racemates, tautomers, or optical isomers thereof.
[0078] In certain embodiments, sPLA2 inhibitors for use in the current
invention are substituted tricyclics having the structure:
R2 COR,
5\ 4
~ %R2i
Z 2
8
R3 N
1
CH2R4
wherein:
Z is selected from the group consisting of cyclohexenyl and phenyl;
R21 is a non-interfering substituent;
R, is -NHNH2 or -NH2;
77
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
R2 is selected from the group consisting of -OH and -O(CH2) ,,,R5; wherein R5
is selected from the group consisting of H, -CO2H, -CONH2, -C02(C1 -C4
alkyl), -SO3H,-SO3(C1-C4 alkyl), tetrazolyl, -CN, -NH2, -NHSO2R15, -
CONHSO2R,5, phenyl, phenyl substituted with -CO2H or -CO2(C,-C4)alkyl,
and
0
I I (RsR7)
wherein R6 and R7 are each independently selected from the group consisting
of -OH, -O(Cl-C4)alkyl; Rj;, is selected from the group consisting of -(Cl-
C6)alkyl and -CF3; and m is 1-3;
R3 selected from the group consisting of H, -O(C,-C4)alkyl, halo, -(C,-
C6)alkyl, phenyl, -(C,-C4)alkylphenyl, phenyl substituted with -(Cl-C6)alkyl,
halo, or -CF3, -CHZOSi(C1-C6)alkyl, furyl, thiophenyl, -(C,-C6)hydroxyalkyl,
and -(CH2) õRp,; wherein R8 is selected from the group consisting of H, -
CONH2, -NRyRja, -CN, and phenyl; R9 and R,o are each independently
selected from the group consisting of H, -CF3, phenyl, -(Cl-C4)alkyl, -(C,-
C4)alkylphenyl, and -phenyl(C,-C4)alkyl; and n is 1 to 8;
R4 is selected from the group consisting of H, -(C5-C,4)aikyl, -(C3-
C14)cycloalkyl, pyridyl, phenyl, phenyl substituted with -(Cl-C6)alkyl, halo, -
CF3, -OCF3, -(C,-C4)alkoxy, -CN, -(C,-C4)alkylthio, -phenyi(C1-C4)alkyl, -
(Cl-C4)alkylphenyl, phenyl, phenoxy and naphthyl;
and pharmaceutically acceptable salts, solvates, prodrug derivatives,
racemates, tautomers, or optical isomers thereof.
[0079] In certain embodiments, sPLA2 inhibitors for use in the current
invention are selected from the group consisting of: {9-[(phenyl)methyl]-5-
78
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
carbamoylcarbazol-4-yl}oxyacetic acid; 9-benzyl-5,7-dimethoxy-1,2,3,4-
tetrahydrocarbazole-4-carboxylic acid hydrazide; 9-benzyl-5,7-dimethoxy-
1,2,3,4-tetrahydrocarbazole-4-carboxamide; [9-benzyl-4-carbamoyl-7-
methoxy-1,2,3,4-tetrahydrocarbazol-5-yl]oxyacetic acid; [9-benzyl-4-
carbamoyl-7-methoxycarbazol-5-yl]oxyacetic acid; methyl [9-benzyl-4-
carbamoyl-7-methoxycarbazol-5-yl]oxyacetic acid; 9-benzyl-7-methoxy-5-
cyanomethyloxy-1,2,3,4-tetrahydrocarbazole-4-carboxamide; 9-benzyl-7-
methoxy-5-(1 H-tetrazol-5-yi-methyl)oxy)-1,2,3,4-tetrahydrocarbazole-4-
carboxamide; {9-[(phenyl)methyl]-5-carbamoyl-2-methyl-carbazol-4-
yl}oxyacetic acid; {9-[(3-fluorophenyl)methyl]-5-carbamoyl-2-methylcarbazoi-
4-yl}oxyacetic acid; {9-[(3-methylphenyl)methyl]-5-carbamoyl-2-
methylcarbazol-4-yl}oxyacetic acid; {9-[(phenyl)methyl]-5-carbamoyl-2-(4-
trifluoromethylphenyl)-carbazol-4-yl}oxyacetic acid; 9-benzyl-5-(2-
methanesulfonamido)ethyloxy-7-methoxy-1,2,3,4-tetrahydrocarbazole-4-
carboxamide; 9-benzyl-4-(2-methanesulfonamido)ethyloxy-2-
methoxycarbazole-5-carboxamide; 9-benzyl-4-(2-
trifluoromethanesulfonamido)ethyloxy-2-methoxycarbazole-5-carboxamide; 9-
benzyl-5-methanesulfonamidoylmethyloxy-7-methoxy-1,2,3,4-
tetrahydrocarbazole-4-carboxamide; 9-benzyl-4-
methanesulfonamidoylmethyloxy-carbazole-5-carboxamide; [5-carbamoyl-2-
pentyl-9-(phenylmethyl)carbazol-4-yl]oxyacetic acid; [5-carbamoyl-2-(1-
methylethyl)-9-(phenylmethyl)carbazol-4-yl]oxyacetic acid; [5-carbamoyl-9-
(phenylmethyl)-2-[(tri(-1-methylethyl)silyl)oxymethyl]carbazol-4-yl]oxyacetic
acid; [5-carbamoyl-2-phenyl-9-(phenylmethyl)carbazol-4-yl]oxyacetic acid; [5-
carbamoyl-2-(4-chlorophenyl)-9-(phenylmethyl)carbazol-4-yl]oxyacetic acid;
79
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
[5-carbamoyl-2-(2-furyl)-9-(phenylmethyl)carbazol-4-yi]oxyacetic acid; [5-
carbamoyl-9-(phenylmethyl)-2-[(tri(-1-methylethyl)silyl)oxymethyl]carbazol-4-
yl]oxyacetic acid; {9-[(2-Fluorophenyl)methyl]-5-carbamoylcarbazol-4-
yl}oxyacetic acid; {9-[(2-trifluoromethylphenyl)methyl]-5-carbamoylcarbazol-4-
yl}oxyacetic acid; {9-[(2-benzylphenyl)methyl]-5-carbamoylcarbazol-4-
yl}oxyacetic acid; {9-[(1-naphthyl)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic
acid; {9-[(2-cyanophenyl)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid; {9-
[(3-cyanophenyl)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid; {9-[(3,5-
dimethylphenyl)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid; {9-[(3-
iodophenyi)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid; {9-[(2-
Chlorophenyi)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid; {9-[(213-
difluorophenyl)methyl]-5-carbamoylcarbazol--4-yl}oxyacetic acid; {9-[(2,6-
difluorophenyi)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid; {9-[(2,6-
dichlorophenyl)methyi]-5-carbamoylcarbazol-4-yl}oxyacetic acid; {9-[(2-
biphenyl)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid; {9-[(2-
8iphenyl)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid methyl ester; [9-
Benzyl-4-carbamoyl-1,2,3,4-tetrahydrocarbazol-5-yl]oxyacetic acid; {9-[(2-
Pyridyl)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid; {9-[(3-
Pyridyl)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid; [9-benzyl-4-
carbamoyl-8-methyl-1, 2, 3,4-tetrahydrocarbazol-5-y1]oxyacetic acid; [9-benzyl-
5-carbamoyl-l-methylcarbazol-4-yl]oxyacetic acid; [9-benzyl-4-carbarnoyl-8-
fluoro-12,3,4-tetrahydrocarbazol-5-yl]oxyacetic acid; [9-benzyl-4-carbamoyl-
8-chloro-12,3,4-tetrahydrocarbazol-5-yl]oxyacetic acid; [5-carbamoyl-9-
(phenylmethyl)-2-[[(propen-3-yi)oxy] methyl]carbazol-4-yl]oxyacetic acid; [5-
carbamoyl-9-(phenylmethyl)-2-[(propyloxy)methyl]carbazol-4-yl]oxyacetic
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
acid; 9-benzyl-7-methoxy-5-((carboxamidomethyl)oxy)-1,2,3,4-
tetrahydrocarbazole-4-carboxamide; 9-benzyl-7-methoxy-5-cyanomethyloxy-
carbazole-4-carboxamide; 9-benzyl-7-methoxy-5-((1 H-tetrazol-5-yl-
methyl)oxy)-carbazole-4-carboxamide; 9-benzyl-7-methoxy-5-
((carboxamidomethyl)oxy)-carbazole-4-carboxamide; [9-Benzyl-4-carbamoyl-
1,2,3,4-tetrahydrocarbazole-5-yl]oxyacetic acid; {9-[(phenyi)methyl]-5-
carbamoyl-2-methyl-carbazol-4-yi}oxyacetic acid; {9-[(3-fluorophenyl)methyl]-
5-carbamoyl-2-methylcarbazol-4-yl}oxyacetic acid; {9-[(3-
methylphenyl)methyl]-5-carbamoyl-2-methylcarbazol-4-yl}oxyacetic acid; {9-
[(phenyl)methyl]-5-carbamoyl-2-(4-trifluoromethylphenyl)-carbazol-4-
yl}oxyacetic acid; 9-benzyl-5-(2-methanesuifonamido)ethyloxy-7-methoxy-
1,2,3,4-tetrahydrocarbazole-4-carboxamide; 9-benzyl-4-(2-
methanesulfonamido)ethyloxy-2-methoxycarbazole-5-carboxamide; 9-benzyl-
4-(2-trifluoromethanesulfonamido)ethyloxy-2-methoxycarbazole-5-
carboxarnide; 9-benzyl-5-methanesulfonamidoylmethyloxy-7-methoxy-1,2,3,4-
tetrahydrocarbazole-4-carboxamide; 9-benzyl-4-
methanesulfonamidoylmethyloxy-carbazole-5-carboxamide; [5-carbamoyl-2-
pentyl-9-(phenylmethyl)carbazol-4-yl]oxyacetic acid; [5-carbamoyl-2-(1-
methylethyl)-9-(phenylmethyl)carbazol-4-yl]oxyacetic acid; [5-carbamoyi-9-
(phenylmethyl)-2-[(tri(-1-methylethyl)silyl)oxymethyl]carbazol-4-yl]oxyacetic
acid; [5-carbarnoyl-2-phenyl-9-(phenylmethyl)carbazol-4-yl]oxyacetic acid; [5-
carbamoyl-2-(4-chIorophenyl)-9-(phenylmethyl)carbazo1-4-yl]oxyacetic acid;
[5-carbamoyl-2-(2-furyl)-9-(phenylmethyl)carbazoi-4-yi]oxyacetic acid; [5-
carbamoyi-9-(phenylmethyl)-2-[(tri(-1-methylethyl)silyl)oxymethyl]carbazol-4-
yl]oxyacetic acid; {9-[(3-fluorophenyl)methyl]-5-carbamoylcarbazol-4-
81
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
yI}oxyacetic acid; {9-[(3-chlorophenyl)methyl]-5-carbamoylcarbazol-4-
yl}oxyacetic acid; {9-[(3-phenoxyphenyl)methyl]-5-carbamoylcarbazol-4-
yl}oxyacetic acid; {9-[(2-Fluoropheny!)methyl]-5-carbamoylcarbazol-4-
yl}oxyacetic acid; (9-[(2-trifluoromethylphenyl)methyl]-5-carbamoylcarbazol-4-
yl}oxyacetic acid; {9-[(2-benzylphenyl)methyl]-5-carbamoylcarbazol-4-
y1}oxyacetic acid; {9-[(3-trifluoromethylphenyi)methyl]-5-carbamoylcarbazol-4-
yI}oxyacetic acid; {9-[(1-naphthyl)methyl]-5-carbamoylcarbazol-4-yi}oxyacetic
acid; {9-[(2-cyanophenyl)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid; {9-
[(3-cyanophenyl)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid; {9-[(2-
methylphenyl)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid; {9-[(3-
methylphenyl)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid; {9-[(3,5-
dimethylphenyl)methyl]-5-carbamoylcarbazoi-4-yl}oxyacetic acid; {9-[(3-
iodophenyl)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid; {9-[(2-
Chlorophenyi)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid; {9-[(2,3-
difluorophenyl)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid; {9-[(2,6-
difluorophenyl)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid; {9-[(2,6-
dichlorophenyl)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid; {9-[(3-
trifluoromethoxyphenyl)methyl]-5-carbamoylcarbazol-4-yi}oxyacetic acid; {9-
[(2-biphenyl)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid; {9-[(2-
Siphenyl)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid methyl ester; [9-
Benzyl-4-carbamoyl-1,2,3,4-tetrahydrocarbazole-5-yl]oxyacetic acid; {9-[(2-
Pyridyl)methyi]-5-carbamoylcarbazol-4-yl}oxyacetic acid; {9-[(3-
Pyridyl)methyl]-5-carbamoylcarbazol-4-yl}oxyacetic acid; [9-benzyl-4-
carbamoyl-8-methyl-1, 2, 3,4-tetrahydrocarbazol-5-yl]oxyacetic acid; [9-benzyl-
5-carbamoyl-l-methylcarbazol-4-yl]oxyacetic acid; [9-benzyl-4-carbamoyl-8-
82
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
fluoro-1,2,3,4-tetrahydrocarbazol-5-yl]oxyacetic acid; [9-benzyl-5-carbamoyl-
1-fluorocarbazol-4-yl]oxyacetic acid; [9-benzyl-4-carbamoyl-8-chloro-1,2,3,4-
tetrahydrocarbazol-5-yl]oxyacetic acid; [9-benzyl-5-carbamoyl-1-
chlorocarbazol-4-yi]oxyacetic acid; [9-[(Cyclohexyl)methyl]-5-
carbamoylcarbazol-4-yl]oxyacetic acid; [9-[(Cyclopentyl)methyl]-5-
carbamoylcarbazol-4-yl]oxyacetic acid; [5-carbamoyl-9-(phenylmethyl)-2-(2-
thienyl)carbazol-4-yl]oxyacetic acid; [5-carbamoyl-9-(phenylmethyl)-2-
[[(propen-3-yl)oxy]methyi]carbazol-4-yl]oxyacetic acid; [5-carbamoyi-9-
(phenyimethyl)-2-[(propyloxy)methyl]carbazol-4-yl]oxyacetic acid; 9-benzyl-7-
methoxy-5-((carboxamidomethyl)oxy)-1,2,3,4-tetrahydrocarbazole-4-
carboxamide; 9-benzyl-7-methoxy-5-cyanomethyloxy-carbazole-4-
carboxamide; 9-benzyi-7-methoxy-5-((1H-tetrazol-5-yl-methyl)oxy)-carbazole-
4-carboxamide; 9-benzyl-7-methoxy-5-((carboxamidomethyl)oxy)-carbazole-
4-carboxamide; [9-Benzyl-4-carbamoyl-1,2,3,4-tetrahydrocarbazole-5-
yl]oxyacetic acid; (R,S)-(9-benzyl-4-carbarnoyl-1-oxo-3-thia-12,3.4-
tetrahydrocarbazol-5-yl)oxyacetic acid; (R, S)-(9-benzyl-4-carbamoyl-3-thia-
1,2,3,4-tetrahydrocarbazol-5-yl)oxyacetic acid; 2-(4-oxo-5-carboxamido-9-
benzyl-9H-pyrido[3,4-b]indolyl)acetic acid chloride; [N-benzyl-1-carbamoyl-I-
aza-1,2,3,4-tetrahydrocarbazol-8-yl]oxyacetic acid; 4-methoxy-6-
methoxycarbonyl-10-phenylmethyl-6,7,8,9-tetrahydropyrido[1,2-a]indole; (4-
carboxamido-9-phenylmethyl-4,5-dihydrothiopyrano[3,4-b]indol-5-yl)oxyacetic
acid; 3,4-dihydro-4-carboxamidol-5-methoxy-9-phenylmethylpyrano[3,4-
b]indole; 2-[(2,9 bis-benzyl-4-carbamoyl-1,2,3,4-tetrahydro-betacarbolin-5-
yl)oxy]acetic acid; 2-[4-oxo-5-carboxamido-9-(2-methylbenzyl)-9H-pyrido[3,4-
b]indoly!]acetic acid; 2-[4-oxo-5-carboxamido-9-(3-methylbenzyl)-9H-
83
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-(4-methylbenzyl)-
9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-(4-tert-
butylbenzyl)-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-
pentafluorobenzyl-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-
carboxamido-9-(2-fluorobenzyl)-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-
5-carboxamido-9-(3-fluorobenzyl)-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-
oxo-5-carboxamido-9-(4-fiuorobenzyl)-9H-pyrido[3,4-b]indoiyl]acetic acid; 2-
[4-oxo-5-carboxamido-9-(2,6-difluorobenzyl)-9H-pyrido[3,4-b]indolyl]acetic
acid; 2-[4-oxo-5-carboxamido-9-(3,4-difluorobenzyl)-9H-pyrido[3,4-
b]indolyi]acetic acid; 2-[4-oxo-5-carboxamido-9-(2,5-difluorobenzyl)-9H-
pyrido[3,4-b]indolyi]acetic acid; 2-[4-oxo-5-carboxamido-9-(3,5-
difluorobenzyl)-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-
(2,4-difluorobenzyl)-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-
carboxamido-9-(2,3-difluorobenzyl)-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-
oxo-5-carboxamido-9-[2-(trifluoromethyl)benzyl]-9H-pyrido[3,4-b]indolyl]acetic
acid; 2-[4-oxo-5-carboxamido-9-[2-(trifluoromethyl)benzyl]-9H-pyrido[3,4-
b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-[3-(trifluoromethyl)benzyl]-9H-
pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-[4-
(trifluoromethyl)benzyl]-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-
carboxamido-9-[3,5-bis(trifluoromethyl)benzyl]-9H-pyrido[3,4-b]indolyl]acetic
acid; 2-[4-oxo-5-carboxamido-9-[2,4-bis(trifluoromethyl)benzyl]-9H-pyrido[3,4-
b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-(a-methylnaphthyl)-9H-
pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-(b-methylnaphthyl)-
9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-(3,5-
dimethylbenzyl)-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-
84
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
9-(2,4-dimethylbenzyl)-9H-pyrido[3,4-b]indolyljacetic acid; 2-[4-oxo-5-
carboxamido-9-(2-phenylbenzyl)-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-
5-carboxamido-9-(3-phenylbenzyl)-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-
oxo-5-carboxamido-9-(4-phenylbenzyl)-9H-pyrido[3,4-b]indolyl]acetic acid; 2-
[4-oxo-5-carboxamido-9-(1-fluorenylmethy)-9H-pyrido[3,4-b]indolyl]acetic
acid; 2-[4-oxo-5-carboxamido-9-(2-fluoro-3-methylbenzyl)-9H-pyrido[3,4-
bjindolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-(3-benzoylbenzyl)-9H-
pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-(2-phenoxybenzyl)-
9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-(3-
phenoxybenzyi)-9H-pyrido[3,4-bjindolyl]acetic acid; 2-[4-oxo-5-carboxamido-
9-(4-phenoxybenzyl)-9H-pyrido[3,4-b]indolyljacetic acid; 2-[4-oxo-5-
carboxamido-9-[3-[2-(fluorophenoxy)benzyi]]-9H-pyrido[3,4-b]indolyljacetic
acid; 2-[4-oxo-5-carboxamido-9-[3-[4-(fluorophenoxy)benzyl]]-9H-pyrido[3,4-
b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-[2-fluoro-3-
(trifluoromethyl)benzyl]-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-
carboxamido-9-[2-fluoro-4-(trifluoromethyl)benzyl]-9H-pyrido[3,4-
b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-[2-fluoro-5-
(trifluoromethyl)benzylj-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-
carboxamido-9-[3-fluoro-5-(trifluoromethyl)benzyi]-9H-pyrido[3,4-
b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-[4-fluoro-2-
(trifluoromethyl)benzylj-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-
carboxamido-9-[4-fluoro-3-(trifluoromethyl)benzyl]-9H-pyrido[3,4-
b]indolyljacetic acid; 2-[4-oxo-5-carboxamido-9-[2-fluoro-6-
(trifluoromethyl)benzyij-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-
carboxamido-9-(2,3,6-trifluorobenzyl)-9H-pyrido[3,4-b]indolyl]acetic acid; 2-
[4-
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
oxo-5-carboxamido-9-(2,3,5-trifluorobenzyl)-9H-pyrido[3,4-b]indolyl]acetic
acid; 2-[4-oxo-5-carboxamido-9-(2,4,5-trifluorobenzyl)-9H-pyrido[3,4-
b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-(2,4,6-trifluorobenzyl)-9H-
pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-(2,3,4-
trifluorobenzyl)-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-
(3,4,5-trifluorobenzyl)-9H-pyrido[3,4-b]indolylJacetic acid; 2-[4-oxo-5-
carboxamido-9-[3-(trifluoromethoxyl)benzyl]-9H-pyrido[3,4-b]indolyl]acetic
acid; 2-[4-oxo-5-carboxamido-9-[4-(trifluoromethoxyl)benzyl]-9H-pyrido[3,4-
b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-[4-
methoxy(tetrafluoro)benzyl]-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-
carboxamido-9-(2-methoxybenzyl)-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-
oxo-5-carboxamido-9-(3-methoxybenzy!)-9H-pyrido[3,4-b]indolyl]acetic acid;
2-[4-oxo-5-carboxamido-9-(4-methoxybenzyl)-9H-pyrido[3,4-b]indolyl)acetic
acid; 2-(4-oxo-5-carboxamido-9-(4-ethylbenzyl)-9H-pyrido[3,4-b]indolyl]acetic
acid; 2-[4-oxo-5-carboxamido-9-(4-isopropylbenzyl)-9H-pyrido[3,4-
b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-(3,4,5-trimethoxybenzyl)-9H-
pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-(3,4-
methylenedioxybenzyl)-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-
carboxamido-9-(4-methoxy-3-methylbenzyl)-9H-pyrido[3,4-b]indolyl]acetic
acid; 2-[4-oxo-5-carboxamido-9-(3,5-dimethoxybenzyl)-9H-pyrido[3,4-
b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-(2,5-dimethoxybenzyl)-9H-
pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-(4-ethoxybenzyl)-
9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-carboxamido-9-
(cyclohexylmethyl)-9H-pyrido[3,4-b]indolyi]acetic acid; 2-[4-oxo-5-
carboxamido-9-(cyclopentyimethyl)-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-
86
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
oxo-5-carboxamido-9-ethyl-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-
carboxamido-9-(1-propyl)-9H-pyrido[3,4-b]indolyi]acetic acid; 2-[4-oxo-5-
carboxamido-9-(2-propyl)-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-
carboxamido-9-(1-butyl)-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-
carboxamido-9-(2-butyl)-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-
carboxarnido-9-isobutyl-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-oxo-5-
carboxamido-9-[2-(1-phenylethyl)]-9H-pyrido[3,4-b]indolyl]acetic acid; 2-[4-
oxo-5-carboxamido-9-[3-(1-phenylpropyl)]-9H-pyrido[3,4-b]indolyi]acetic acid;
2-[4-oxo-5-carboxamido-9-[4-(1-phenylbutyl)]-9H-pyrido[3,4-b]indolyl]acetic
acid; 2-[4-oxo-5-carboxamido-9-(1-pentyl)-9H-pyrido[3,4-b]indolyl]acetic acid;
2-[4-oxo-5-carboxamido-9-(1-hexyl)-9H-pyrido[3,4-b]indolyl]acetic acid; 4-[(9-
benzyl-4-carbamoyl-1,2,3,4-tetrahydrocarbazol-6-yl)oxy]butyric acid; 3-[(9-
benzyl-4-carbamoyl-1,2,3,4-tetrahydrocarbazol-6-yl)oxy]propylphosphonic
acid; 2-[(9-benzyl-4-carbamoyl-1,2,3,4-tetrahydrocarbazol-6-
yl)oxy]methylbenzoic acid; 3-[(9-benzyl-4-carbamoyl-7-n-octyl-1,2,3,4-
tetrahydrocarbazol-6-yl)oxy]propylphosphonic acid; 4-[(9-benzyl-4-carbamoyl-
7-ethyl-1,2,3,4-tetrahydrocarbazol-6-yl)oxy]butyric acid; 3-[(9-benzyl-4-
carbamoyl-7-ethyl-12,3,4-tetrahydrocarbazol-6-yl)oxy]propylphosphonic acid;
3-[(9-benzyl-4-carbamoyl-7-ethyl-1, 2, 3,4-tetra hydrocarbazol-6-
yl)oxy]propylphosphonic acid; (S)-(+)-4-[(9-benzyl-4-carbamoyl-7-ethyl-
1,2,3,4-tetrahydrocarbazol-6-yl)oxy]butyric acid; 4-[9-benzyl-4-carbamoyl-6-
(2-cyanoethyl)-1,2,3,4-tetrahydrocarbazol-6-yl]oxybutyric acid; 4-[9-benzyl-4-
carboxarnido-7-(2-phenylethyl)-1,2,3,4-tetrahydrocarbazol-6-yl]oxybutyric
acid; 4-[9-benzyl-4-carboxamidocarbazol-6-yl]oxybutyric acid; methyl 2-[(9-
benzyl-4-carbamoyl-1,2,3,4-tetrahydrocarbazol-6-yl)oxy]methylbenzoate; 4-[9-
87
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
benzyl-4-carbamoyl-7-(2-cyanoethyl)-1, 2, 3,4-tetrahydrocarbazol-6-
yl]oxybutyric acid; 9-benzyl-7-methoxy-5-cyanomethyloxy-1,2,3,4-
tetrahydrocarbazole-4-carboxamide; [9-benzyl-4-carbamoyl-8-methyl-
carbazole-5-yl]oxyacetic acid; and [9-benzyl-4-carbarnoyi-carbazole-5-
yl]oxyacetic acid, or pharmaceutically acceptable salts, soivates, prodrug
derivatives, racemates, tautomers, or optical isomers thereof.
[0080] In certain preferred embodiments, an sPLA2 inhibitor for use in the
present invention is ((3-(2-Amino-1,2-dioxoethyl)-2-ethyl-1-(phenylmethyl)-1 H-
indol-4-yl)oxy)acetic acid, also referred to herein as compound A-001.
Compound A-001, which is also referred to in the art as S-5920 or LY315920,
has the structure:
0
O NHZ
HO
,--._
O
N
CH3
A-001 competitively inhibits sPLA2.
[0081] In certain other preferred embodiments, an sPLA2 inhibitor for use in
the present invention is [[3-(2-Amino-1.2-dioxoethyi)-2-ethyi-1-(phenylmethyl)-
1 H-indol-4-yl]oxy]acetic acid methyl ester, also referred to herein as
compound A-002, Compound A-002 has the structure:
88
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
O
O NH2
i-{ aC ~ O
O
N
C
CH3
Compound A-002, which is sometimes referred to in the art as S-3013 or
LY333013, is a prodrug form of A-001 that is rapidly absorbed and hydrolyzed
to A-001 following administration to a subject_
[0082] In certain other preferred embodiments, an sPLA2 inhibitor for use in
the present invention is {9-[(phenyl)methyl]-5-carbamoylcarbazol-4-
yI}oxyacetic acid, also referred to herein as compound A-003 or LY433771.
Compound A-003 has the structure:
O O NH2
O
+Na-O
N
[0083] In still other preferred embodiments, an sPLA2 inhibitor for use in the
present invention is ((3-(2-amino-1,2-dioxoethyl)-2-mthyl-l-(phenylmethyl)-
89
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
1 H-indol-4-yl)oxy)acetic acid N-morpholino ethyl ester, also referred to
herein
as compound 421079. Compound 421079 has the structure:
0
N 0
O
NH
cw
N
Like A-002, compound 421079 is a prodrug of A-001.
[0084] The following examples are provided to better illustrate the claimed
invention and are not to be interpreted as limiting the scope of the
invention.
To the extent that specific materials are mentioned, it is merely for purposes
of illustration and is not intended to limit the invention. One skilled in the
art
may develop equivalent means or reactants without the exercise of inventive
capacity and without departing from the scope of the invention. It will be
understood that many variations can be made in the procedures herein
described while still remaining within the bounds of the present invention. It
is
the intention of the inventors that such variations are included within the
scope of the invention.
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
Examples
Example 1: Effect of A-002 administration on plasma cholesterol levels and
atherosclerotic plaque formation in mice:
[0085] Male ApoE-'- mice were fed a high fat diet (21 % fat, 0.15 /o
cholesterol, 19.5% casein) ad libitum for two weeks in order to acclimate to
the diet. Plasma cholesterol levels and body weight were measured in order
to obtain baseline levels, and mice were randomized into three groups of 20
mice based on these measurements. After the acclimation period, mice
remained on the high fat diet and were administered A-002 (30 mg/kg or 90
mg/kg) or vehicle only (5% acacia) twice a day for 16 weeks by oral gavage.
[0086] Plasma cholesterol levels and body weight were measured at 4, 8, 127
and 16 weeks after the start of A-002 administration. Comparisons of
measurernents between test and control mice at each time period were
performed using a two-way analysis of variance test (ANOVA) for repeated
measures, followed by post-hoc Bonferroni test for significance.
[0087] At the end of the 16th week, mice were sacrificed and plasma
samples, heart tissue, and aortas from the heart to approximately 3 mm distal
to the iliac bifurcation were collected. Aortas were placed on microscope
slides and stained with Oil Red 0 for scanning and image analysis of
atherosclerotic lesion size. Vessel images were captured using a Microtek
Scanmaker 9600XL scanner (Microtek, Carson, CA) and Photoshop 6.0
software (Adobe Systems Inc., San Jose, CA). Atherosclerotic plaque
coverage was quantified for the entire length of the vessel including the arch
using Image-Pro Plus software. Vessels were also analyzed for plaque
coverage in the descending abdominal aorta, starting in the region of the
first
91
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
intercostal branches to the iliac bifurcation (not including the arch). Plaque
content was expressed as a percent of the aortic surface. For assessment of
significance in atherosclerotic plaque content between test and control mice,
a
one-way analysis of variance test followed by Bonferroni post-hoc test was
used.
[0088] All three groups of mice (control, 30 mg/kg A-002, and 90 mg/kg A-
002) had similar baseline body weights (Figure 1). Body weight increased
over the 16 week test period by approximately 155%, 150%, and 141 % for
control, 30 mg/kg A-002, and 90 mg/kg A-002 mice, respectively (Figure 1)_
There was no statistically significant difference in body weight between the
three groups.
[0089] Baseline total plasma cholesterol levels were not significantly
different
between the three groups (Figure 2). At four weeks, mice receiving A-002 at
either dosage exhibited a significant decrease in total cholesterol. This
effect
remained consistent throughout the remainder of the 16 week test period. At
16 weeks, the change from baseline in total cholesterol was +15%, -10%, and
-12% in control, 30 mg/kg A-002, and 90 mg/kg A-002 mice, respectively
(Figure 2). There was no apparent dose-response effect.
[0090] Control mice exhibited approximately 13% plaque coverage in aortic
tissue at 16 weeks (Figure 3). 30 mg/kg A-002 and 90 mg/kg A-002 mice
exhibited 6.3% and 6.8% plaque coverage, respectively (Figure 3).
Therefore, A-002 treatment resulted in a significant decrease in plaque
content.
Example 2: Effect of A-002 administration on an iq otensin II-mediated
atherosclerotic plaque forrnation and aneurysm:
92
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
[0091] A mouse model of accelerated atherosclerosis was utilized to
determine the effect of A-002 administration on atherosclerosis and aneurysm
formation. ApoE-/- mice acclimated to the same high fat diet utilized in
Example 1 were administered angiotensin II with water, angiotensin with 5%
acacia, or saline infusion and water twice daily for four weeks. Angiotensin
11
has been shown to promote atherosclerosis and aneurysm formation in apoE
deficient mice (Daugherty 2000)- At the end of the test period, plaque
coverage was assessed by three independent reviewers, and their
assessments were averaged to determine plaque coverage.
[0092] Angiotensin II with water or with 5% acacia resulted in similar rates
of
plaque coverage (Figure 4) and aneurysm (Table 1). Administration of A-002
at 30 mg/kg twice daily significantly decreased plaque content and aortic
level
aneurysms in angiotensin I1/acacia mice (Figure 4).
Table 1: Effect of A-002 administration on aneurysm rate:
Treatment group Aneurysm rate {%)
--------_~._ __-- -- ------------ ------
Angiotensin II + 5% acacia 22% (2/9)
--------- --- ----------- ----------------- -----
Angiotensin + water 25% (5/20)
Saline + water 0 /o (0/25)
Angiotensin II + 5% acacia + 30 0% (0/16)
mg/kg A-002
Example 3: Effect of A-002 or A-002 plus statin on lesion formation and
composition in mice:
[0093] Body weight, plasma total cholesterol, and plasma HDL levels were
measured for six groups of twelve ApoE-/- mice. The mice were then placed
on a high fat diet for twelve weeks to allow for formation of significant
fatty
atherosclerotic plaques. The high fat diet was a modified version of the
Western diet (TD.88137. Harlan Teklad, Madison, WI) consisting of casein
(195 g/kg), DL-methionine (3 g/kg), sucrose (341.44 g/kg), corn starch (150
93
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
g/kg), anhydrous milkfat (210 g/kg), cholesterol (1.5 g/kg), cellulose (50
g/kg),
mineral mix AIN-76 (Tekiad; 35 g/kg), calcium carbonate (4 g/kg), vitamin mix
(Teklad #40060; 10 g/kg), ethyoxyquin (antioxidant, 0.04 g/kg), and either
0,0156 g/kg ("low dose") A-002 (Group A), 1.56 g/kg ("high dose") A-002
(Group B), 0.02 g/kg pravastatin (Group C), low dose A-002 plus 0.02 g/kg
pravastatin (Group D), high dose A-002 plus 0.02 g/kg pravastatin (Group E),
or vehicle only (Group F). Mice were administered diets A-F in a blinded
manner, and food was provided ad libitum. The diet was initially prepared
based on the assumption that each mouse weighed 22.5 g and ate 4.5 g/day_
Based on body weight and food consumption, the diet was modified over the
12 week course of treatment to adjust for a body weight of 26 g and a food
intake of 2.5 g/day. The dosages of A-002 were selected to cover a range of
100-fold based on pharmacokinetics in ApoE-/- mice as well as toxicology and
previously observed efficacy in this model.
[0094] After twelve weeks, mice were sacrificed and plasma, heart tissue,
and aortic tissue were collected. En face lesion size was deterrnined via
digital imaging analysis. Percent lesion coverage for each dosage group is
summarized in Table 2 and Figure 5. Plaque content (as measured as en
face lesion) was reduced in mice administered A-002 alone, statin alone, or A-
002 in combination with statin. The reduction in plaque content in mice
administered A-002 plus statin was substantially greater than the reduction in
mice administered either compound alone (Table 2, compare Group D vs.
Groups A and C; Group E vs_ Groups B and C). Further, the reduction in
plaque content in mice administered A-002 plus statin was substantially
greater than the sum of the reduction in plaque content in mice administered
94
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
A-002 alone and mice administered statin alone. (Table 2, compare the
change in mean % en face lesion vs. control for Group D (i.e., -4.210%) with
the sum of the change in mean % en face lesion vs. control for Group A and
Group C (i.e., the sum of -2.241 % and -0.813%, or -3.054%).) Thus, the
administration of A-002 plus statin reduces plaque content in a synergistic
manner.
Table 2: Effect of A-002 and/or statin administration on en face lesion size:
Dosage # of mice Mean % en Change in P-value vs.
group face lesion mean % en control
face lesion
vs. control Low A-002 11 3.386 -2.241 0.0597
(A)
High A-002 12 1.469 -4.158 0.0006
(B)
Statin 12 4.814 -0.813 0.4797
(C)_
Low A-002 10 1.417 -4.210 0.0008
plus statin
(D)
High A-002 10 0.774 4.853 0.0001
plus statin
(E)
- -------
Control 12 5.627 -- --
(F) - - -
[0095] Plasma total cholesterol and HDL levels were measured. Mean levels
of total cholesterol were decreased in mice administered A-002 alone or in
combination with statin (Table 3, Figure 6). Adrninistration of high dosage A-
002 plus statin resulted in a greater decrease in total cholesterol than
administration of high dosage A-002 or statin alone (Table 3, compare Group
E vs. Groups B and C; Figure 6). Mean levels of HDL were increased in mice
administered A-002 at either dosage (Table 4). This increase was greater in
mice administered high dose A-002 plus statin than in mice administered high
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
dose A-002 or statin alone (Table 4 compare Group E vs. Groups B and C;
Figure 7).
Table 3: Effect of A-002 and/or statin administration on plasma total
cholesterol levels
Dosage # of mice Mean total P-value vs.
group cholesterol control
concentration
(mg/di)
----
Low A-002 11 1355.545 0.2296
(A)
----------------
High A-002 12 1301.750 0.0792
(B)
Statin 12 1405.417 0.4647
(C)
Low A-002 10 1399.700 0.4521
plus statin
(D)
High A-002 10 1219.300 0.0152
plus statin
(E)
---
Control 12 1478.083 --
(F)
Table 4: Effect of A-002 and/or statin administration on plasma HDL levels
Dosage # of mice Mean HDL P-value vs.
group concentration control
{mg/dl)
Low A-002 (A) 11 21.000 <0.0001
High A-002 12 17.167 0.0825
(B)
Statin 12 14.500 0.9067
(C)
Low A-002 -----10---- - -- -----21.300--- ---- <0.0001 plus statin
(D)
._._._. ----...--------------
High A-002 10 18.500 0.0123
plus statin
(E)
Control 12 14.667 --
(F)
Example 4: Effect of A-002 or A-002 plus statin administration on serum lipid
levels in humans:
96
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
[0096] 204 human subjects from the United States with CVD, specifically
stable coronary artery disease, were randomized to receive placebo or A-
002 via oral administration twice a day over an eight week administration
period at dosages of 50 mg, 100 mg, 250 mg, or 500 mg. Levels of various
lipids and inflammatory markers were measured at the outset of the trial and
at the end of weeks four and/or eight. For data sets with a normal
distribution, mean levels of lipids or inflammatory markers were analyzed.
For data sets with non-normal distribution, median levels were analyzed.
Administration of A-002 at all dosages tested resulted in a decrease in mean
levels of serum LDL (Table 5), LDL particles (Table 6), small LDL particles
(Table 7), total cholesterol (Table 8), and TG (Table 9), and median levels of
sPLA2 (Table 10). In addition, administration of A-002 decreased median
levels of CRP (Table 11). The magnitude of the observed decreases in LDL
particle, small LDL particle, and CRP levels are particularly noteworthy
because subjects treated with placebo exhibited increases in these markers
over the course of the trial (Tables 6, 7, and 11).
Table 5: Changes in serum LDL concentration in ITT population following A-
002 administration
A-002 Placebo
Baseline # of 154 37
subjects _
Mean 76.8 mg/dl 77.7 mg/dl
[LDL] Week 8 # of 143 36
subjects
observed
Change in -8.2 mg/dl -1.5 mg/dl
mean
[LDL] vs.
baseline
% change -10.68% -1.93%
vs.
97
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
---
baseline
p-value <0.0001 0.5456
vs.
baseline
Table 6: Changes in serum LDL particle concentration in ITT population
following A-002 administration
A-002 Placebo
--
Baseline # of 158 38
subjects
_.-----------
Mean 1031.6 nmol/L 1004.8
[LDL nmol/L
particle]
Week 8 # of 129 33
subjects
observed
_------------
Change in -73.6 nmol/L +47.7 nmol/L
mean
[LDL
particle]
vs.
baseline
% change -7.13% +4.75%
vs.
baseline
Table 7: Changes in serum small LDL particle concentration in ITT population
following A-002 administration A-002 Placebo
Baseline # of 158 38
subjects
Mean 735.8 nmol/L 661.7 nmol/L
[small
LDL
particle]
Week 8 # of 129 33
subjects
observed
----
Change in 61.3 nmol/L +77.0 nmol/L
mean
[small
LDL
particle]
vs.
baseline
% change 8_33% +11.64%
98
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
C ` baseline ~ --- ~ -- ~
Table 8: Changes in serum total cholesterol concentration in ITT population
following A-002 administration
---
A-002 Piacebo
Baseline #of _--- --~ 158 -------- -_39_..----_
subjects
Mean [total 156.8 mg/dl 156.9 mg/dl
cholesterol]
Week 8 # of 146 37
subjects
observed
Change in -12.8 mg/dl -4.7 mg/dl
mean [total
cholesterol]
vs. baseline
% change -8.16% -2.99%
vs. baseline
- -------___ _-.---.__- ---_.-.__----------- ~
p-value vs. <0.0001 0.1130
baseline
Table 9: Changes in serum TG concentration in ITT population following A-
002 administration
A-002 Placebo
Baseline # of 158 39
subjects
__
Mean [TG] 151.5 mgJd1 50.6 mg/dl
Week 8 # of 146 37
subjects
observed
Change in -10.1 mg/dl -0.7 mg/dl
mean [TG]
vs.
baseline
% change -6.67% -0.46%
vs.
baseline
Table 10: Changes in serum sPLA2 concentration in ITT population following
A-002 administration
A-002 Placebo
._~ -- - --- - _ _- --- _
Baselme # of 159 41
---- isubjects__
99
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
Median 2.30 ng/ml 2.20 ng/ml
[sPLA2]
--._____.~ ------- ___ _____.._.__ ---------------_---_____
Week 8 # of 110 29
subjects
observed
-_-____-----
Change in -2.00 ng/ml -0.30 ng/ml
median
[s P LAzI
vs.
baseline
-----------______.__-_-___-----------
% change -86.96% -13.64%
Vs.
baseline
p-value <0.0001 -- - ----0.0207
vs.
baseline
Table 11: Changes in serum CRP concentration in ITT population following A-
002 administration
-------------
A-002 Plac_ebo
Basel i ne # of 155 ------ ------ 38----
subjects
Median 1.50 mg/L 2.00 mg/L
[CRP]
Week 8 # of 143 36
subjects
observed
Change in -0.20 mg/L +0.10 mg/L
median
[CRP] vs.
baseline
% change -13.33% +5.00%
vs.
baseline
p-value 0.0205 0_4478
vs.
baseline
[0097] The median baseline LDL concentration in the 204 subject United
States ITT population was 72.0 mg/dl. A-002 administration resulted in a
significant decrease in mean LDL levels in a 97 subject subpopulation with
baseline LDL levels equal to or higher than the median concentration (Table
100
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
12). In addition, administration of A-002 decreased mean serum LDL levels in
a 53 subject subpopulation with diabetes (Table 13).
Table 12: Changes in serum LDL concentration in subpopulation with baseline
serum LDL concentration greater than or equal to 72 mg/dl following A-002
administration
------- ------- - A-002 __ _ Placebo
Baseline # of 79 18
subjects
--- -------- _.-----__.-___-
Mean 95.4 mg/di 98.0 mg/dl
[LDL]
Week 8 # of 70 16
subjects
Change in -14.0 mg/di -3.5 mg/dl
mean
[LDL] vs.
baseline
% change -14.68% -3.57%
vs.
baseline
p-value <0.0001 0.3428
vs.
baseline
Table 13: Changes in serum LDL concentration in diabetic subpopulation
following A-002 administration
A-002 Placebo
Baseline # of 43 8
sub[ects
Mean 63.7 mg/di 75.1 mg/dl
[LDL]
Week 8 # of 37 8
subjects
Change in -10.6 mg/di -4.3 mg/di
mean
[LDL] vs.
baseline
% change -16.64 /o -5.73%
vs.
baseline
[0098] A decrease in mean small LDL particle levels was observed in the
subset of the US population that was receiving statin treatment during the
trial
101
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
(Table 14), indicating that administration of A-002 plus statin results in a
greater decrease in small LDL particles than administration of statin alone.
Likewise, a decrease in mean serum LDL levels was observed in an 81
subject subpopulation of the statin treatment group exhibiting elevated
baseline LDL levels (i.e., baseline LDL levels greater than or equal to 72
mg/dl) (Table 15).
Table 14: Changes in serum small LDL particle concentration in statin
subpopulation following A-002 administration
A-002 Placebo
Baseline # of 139 33
sub'ects
Mean 735.3 nmol/L 664.4 nmol/L
[small
LDL
particle]-
Week 8 # of 114 27
subjects
observed
----------__---
Change in -66.2 nmol/L +88.1 nmol/L
mean
[small
LDL
particle]
vs.
baseline
% change -9.00%+13.26%
vs.
baseline
Table 15: Changes in serum LDL concentration in statin subpopulation with
baseline serum LDL concentration greater than or equal to 72 mg/di
following A-002 administration
A-002 Placebo
Baseline # of 66 15
sub'ects
Mean 92.2 mg/dl 86.3 mg/dl
[LDL]
Week 8 # of 58 13
subjects
102
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
Change in -15.0 mg/dl -3.8 mg/dl
mean
[LDL] vs.
baseline
% change -16.27% -4.40%
vs.
baseline
p-value <0.0001 0.3792
vs.
baseline
[0099] The combined effect of A-002 and statin administration on LDL
levels was tested in a population of 332 subjects. This population consisted
of the original 204 United States subjects, plus an additional 128 subjects
from the Ukraine with stable CAD that received placebo or A-002 based
according to the same protocol as the United States subjects. A decrease in
mean serum LDL levels was observed in the statin subpopulation at four and
eight weeks (Table 16), indicating that administration of A-002 and statin
results in a greater decrease in serum LDL levels than administration of
statin alone. Similarly, A-002 decreased LDL levels in subjects receiving a
variety of non-statin compounds used in the treatment of CVD, including
ezetimibe, AEGR-733/ezetimibe, colesevelam hydrochloride (WelChol ),
MK-0524A (Cordaptive ), lisinopril/MC-1 antibody (MC-4232), and
Angiotensin Receptor Blocker (ARB)/MC-1 (MC-4262).
Table 16: Changes in serum LDL concentration in statin subpopulation
following A-002 administration
---- A-002___--- Placebo
Baseline # of 156 36
subjects
Mean 76.9 mg/dl 74.5 mg/dl
[LDL] -~. _--- - -
Week 4# of 152 35
sub'ects
Change in -10.8 mg/dl -2.5 mg/dl
mean
103
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
[LDL] vs.
baseline % change -14.04% -3.36%
vs.
baseline
_----------------
Week 8 # of 131 31
sub'ects
Change in -9.6 mg/dl -1.6 mg/dl
mean
[LDL] vs.
baseline
% change -12.48% -2.15%
vs.
baseline
[00100] Comparison of the above statin subpopulation results with results for
subjects that were not receiving statin treatment indicates that
administration
of A-002 plus statin results in a greater decrease in mean serum small LDL
particle and LDL levels than the expected additive effect of A-002 and statin
(compare Table 17 to Table 14 and Table 18 to Table 16). In the United
States population, mean serum small LDL particle levels were reduced at
eight weeks by 9.00% in subjects administered A-002 plus statin (Table 14),
versus a decrease of 3.25% in subjects administered A-002 alone (Table 17)
and an increase of 13.26% in subjects administered statin alone (Table 14).
These results summarized in Table 20. In the combined United States and
Ukraine populations, mean serum LDL levels were decreased by 14.04% at
week four in subjects administered A-002 plus statin (Table 16), versus a
decrease of 6.93% in subjects administered A-002 alone (Table 18) and a
decrease of 3.36% in subjects administered statin alone (Table 16). At week
eight, mean serum LDL levels were decreased by 12.48% in subjects
administered A-002 plus statin (Table 16), versus a decrease of 4.22% in
subjects administered A-002 alone (Table 18) and a decrease of 2.15% in
subjects adrninistered statin alone (Table 16). These results are also
104
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
summarized in Table 20. Therefore, administration of A-002 plus statin
decreases LDL and small LDL particle levels in a synergistic manner.
[00101] This synergism was also observed in subjects from the United
States population that had baseline LDL concentrations of 72 mg/dl or
greater (compare Table 19 to Table 15). Mean serum LDL levels were
decreased at eight weeks by 16.27% in subjects administered A-002 plus
statin (Table 15), versus a decrease of 8.23% in subjects administered A-
002 alone (Table 19) and a decrease of 4.40% in subjects administered
statin alone (Table 15). These results are summarized in Table 20.
Table 17: Changes in serum small LDL particle concentration in non-statin
subpopulation following A-002 administration
- A-002 Placebo
_------- __ -__.-----
Baseiine # of 19 5
subjects
Mean 739.5 nmol/L 644.4 nmol/L
[smal!
LDL
particle]
Week 8 # of 15 6
subjects
observed
Change in -24.0 nmol/L +19.4 nmol/L
mean
[small
LDL
particle]
vs.
baseline
% change -3.25% +3.01%
vs.
baseline
Table 18: Changes in serum LDL levels in non-statin subpopulation following
A-002 administration
----
A-002 Placebo
-- -- - -.._
Baseline ~ # of 102 21
subjects
105
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
Mean 125.6 mg/dl 135.6 mg/di
[LDL]
Week 4 # of ------- 86 -- - -- 22
sub'ects
Change in -8.7 mg/dl +0.2 mg/dl~-
mean
[LDL] vs.
baseline
% change -6.93% +0.15%
vs.
baseline
_..------ --_ ___ _ ------------- -- -___ ---- _,.----------- ------- -----
Week 8 # of 20 5
sub1'ects Change in -5.3 mg/dl --1.3 mg/dl
mean
[LDL] vs.
baseline
% change -4.22% -0.96%
vs.
baseline
Table 19: Changes in serum LDL concentration in non-statin subpopulation
with baseline serum LDL levels greater than or equal to 72 mg/di following A-
002 administration
__ A-002LL Placebo
Baseline # of i 13 3
sub'ects
Mean 111.8 mg/dl 156.3 mg/dl
[LDL]
Week8 #of ~ 12 3
subjects
-- ____--_--
Change in -9.2 mg/dl -2.3 mg/di
mean
[LDL] vs.
baseline
% change -8.23% -1.47%
vs.
baseline
p-value 0.0574 0.8021
vs.
baseline
Table 20: Summary of the synergistic effects of combined A-002 and statin
administration versus administration of A-002 or statin alone
------- _ __ -- --- - _~.
( Observed Observed_ ~ Expected ~ Observed
106
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
___ . _ . __--- ------_ _._..-- - -._. _ -----._,- ,--- ----------. -_ __ ___-
change change change from change
following following administration following
administration administration of A-002 plus administration
of A-002 alone of statin alone statin of A-002 plus
(change for A- statin
002 alone plus
change for
statin alone)
----
Mean serum -3.25% +13.26 /o +10.01% -9.00%
[small LDL (Table 17) (Table 14) (-3.25% plus (Table 14)
particle] at +13.26%)
week 8
Mean serum -6.93% -3.36% -10.29% -14.04%
[LDL] at week (Table 18) (Table 16) (-6.93% plus (Table 16)
4 -3.36%) Mean serum -4.22% -2.15 -6.37% -12.48
[LDL] at week (Table 18) (Table 16) (-4.22% plus (Table 16)
8 -2.15%
Mean serum -8.23% -4.40% -12.63% -16.27%
[LDL] in _ 72 (Table 19) (Table 15) (-8.23% plus (Table 15)
mgldi -4.40%)
subpopulation
at week 8
[00102] Serum LDL data for subjects in the statin subpopulation were
subdivided based on the specific statin each subject was receiving. Statins
with significant representation included atorvastatin, rosuvastatin,
simvastatin,
lovastatin, pravastatin, and fluvastatin, as well as the statin combination
drugs
simvastatin/ezetimibe (Vytorin(@), atorvastatin/ezetimibe,
atorvastatin/amiodipine (Caduet ), lovastatin/extended release niacin
(Advicor ), rosuvastatin/TriCor , rosuvastatin/ABT-335, simvastatin/extended
release niacin (Simcor ), simvastatin/MK-0524A (MK-0524B),
pravastatin/fenofibrate, atorvastatin/APA-01, and TAK-457/statin. Statin and
statin combination dosages varied within each individual statin subgroup. The
number of test and placebo subjects in each individual statin subgroup frorTi
the larger statin subpopulation was too low to allow for detailed statistical
analysis. However, there appeared to be a trend towards the same
107
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
synergistic decrease in LDL levels that was observed in the statin
subpopulation as a whole in several of the statin subgroups, such as for
example with certain dosages of atorvastatin, rosuvastatin, and simvastatin.
These results indicate that the observed synergy between A-002 and statins
is not limited to a particular statin.
[00903]The synergistic effect of A-002 and compounds used in the treatment
of CVD on LDL levels was not limited to statins. For example, 30 subjects
from the ITT population were receiving ezetimibe at a dosage of 10 mg during
the course of the A-002 trial. Administration of A-002 resulted in a decrease
in mean serum LDL levels in the ezetimibe subpopulation after eight weeks
(Table 21), indicating that administration of A-002 plus ezetimibe results in
a
greater decrease in LDL levels than administration of ezetimibe alone (Table
21). As with the statin subpopulation, administration of A-002 in conjunction
with ezetimibe resulted in a synergistic decrease in mean LDL levels that was
greater than the expected additive effect of A-002 and ezetimibe (Table 22).
Table 21: Changes in serum LDL concentration in ezetimibe subpopulation
following A-002 administration
~ A-002 Placebo
Baseline # of 28 12
subjects
Mean 81.0 mg/dl 79.6 mg/dl
[LDL]__
Week 8 # of 28 12
subjects
------------
Change in -15.4 mg/dl -1.0 mg/dl
mean
[LDL] vs.
baseline % change -17.8% -1.7%
vs.
baseline
108
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
Table 22: Summary of the synergistic effects of combined A-002 and
ezetimibe administration versus administration of A-002 or ezetimibe alone
Observed Observed Expected Observed
change change change from change
following following administration following
administration administration of A-002 plus administration
of A-002 alone of ezetimibe ezetimibe of A-002 plus
alone (change for A- ezetimibe
002 alone plus
change for
_ ezetimibe)
Mean serum -4 22% -1.7% -5.92% -17.8%
[LDL] at week (Table 18) (Table 21) (-4.22% plus (Table 21)
8 -1.7%)
[00104] Serum TG, CRP, and IL-6 levels were measured at eight weeks in 86
subjects from the ITT population diagnosed with metabolic syndrome.
Administration of A-002 resulted in a decrease in serum levels of each of
these markers (Tables 23-25). Subjects administered placebo exhibited an
increase in CRP and IL-6 levels (Tables 24-25).
Table 23: Changes in serum TG concentration in metabolic syndrome
subpopulation following A-002 administration
---
A-002 Placebo
Baseline # of 70 16
subjects
Median 171.5 mg/dl 168.0 mg/dl
[TG] Week 8 # of 63 13
subjects
observed
Change in -24.0 mg/di -3.0 mg/dl
median
[TG] vs.
baseline
% change -13.99 /o -1.79%
vs.
baseline
Table 24: Changes in serum CRP concentration in metabolic syndrome
subpopulation following A-002 administration
109
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
----
A-002 Placebo
Baseline # of 69 16
subects_ _ _ _
Median 2.40 mg/L 2.60 mg/di
[CRP]
W ee k 8 # of -- --`-- --- 62 ----- - 13
subjects
observed _--
Change in -0_3 mg/L +0.1 mg/L
median
[CRP] vs.
baseline
__.~-----_ ---------------- __- ------------__.
% change -12.50% +3.84%
vs.
baseline
Table 25: Changes in serum IL-6 concentration in metabolic syndrome
subpopulation following A-002 administration
A-002 Placebo
Baseline # of 68 16
sub'ects
Mean [IL- 5.60 pg/mi 5.17 pg/mi
6] (15.30) (2.60)
Week 8 # of 44 10
subjects
observed
Change in -0_24 pg/mi +0.29 pg/mi
mean [IL-
6] vs.
baseline % change -4.29% +5.61 /o
vs.
baseline
Example 5: A-002 plus statin combination tablet:
[00105] Fixed dose tablets containing A-002 and one or more statins may be
generated using methods known in the art. For example, a fixed dose tablet
containing a therapeutically effective amount of A-002 (e.g., 250 or 500 mg)
and a therapeutically effective amount of a statin (e.g., 10, 20, 40, or 80
mg),
may be generated using a formula such as that set forth in Table 26. One of
ordinary skill in the art will recognize that additional components may be
110
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
added to this generic formulation. For example, a compound such as calcium
carbonate may be added to the formulation to enhance dissolution and
solubility. Likewise, one of skill in the art will recognize that this
formulation is
just one example of a generic A-002/statin formulation, and that the identity
and weight of the recited components within the formulation niay be varied
without undue experimentation.
Table 26: Generic A-002/statin formulation:
Component Typical weight
percenta
------------ _ ___--------~ _.. A-002 Varies depending on
desired dosage
Statin Varies depending on
desired dosage
Anhydrous lactose 20-50%
Lactose monohydrate 20-50%
H droxyprop I celiulose 2-6%
Croscarmellose sodium 0.5-5%
Polysorbate 80 0.1-3%
Microcrystalline cellulose 5-20%
Magnesium stearate ___ 0.25-3%
[00106] To verify the feasibility of including both A-002 and a statin in a
single
formulation, fixed dose tablets containing 250 mg A-002 and 40 mg
simvastatin (marketed as Zocor ) were generated. Unit and batch formulas
for these tablets are set forth in Tables 27 and 28, respectively. All
inactive
ingredients were purchased from Spectrum Chemicals. Butylated
hydroxyanisole was included in the formulation for consistency with the
i-narketed form of simvastatin (Zocor ).
Table 27: Unit formula for A-002/simvastatin tablets:
Component Function _ mgltablet
A-002 Active inqredient 250
-
------
Simvastatin Active ingredient 40
__-
Anhydrous lactose Diluent 119.18
Lactose fast flo Diluent 59.58
_.
Hydroxypropyl cellulose Binder 115
111
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577 H PC)---
Croscarmellose sodium Disintegrant 24.3
(divided into two equal
portions for inner/extra
granular
Polysorbate 80 Surfactant 0.54
_.-.____
-----------_--_..__. --_~ Antioxidant 0.01%
Butylated
hydroxyanisole (BHA)
Microcrystalline Diluent 70.20
cellulose 200 (MCC)
__-
Ma g nesium stearate Lubricant 2.70
Purified water Solvent --
tablet weight 580 mg
Total
Table 28: Batch formula for A-002/simvastatin tablets:
Component Function glbatch
A-002 Active ingredient 259
Simvastatin Active ingredient 41
-----
Anhydrous lactose Diluent 123 __ ---
Lactose fast flo Diluent 62
Hydroxypropyl cellulose Binder 14
(HPC)
Croscarmellose sodium Disintegrant 6.6/8.4
(divided into two equal
portions for inner/extra
_granuiar
Polysorbate 80 Surfactant
Butylated hydroxanisole Antioxidant 0.1
(BHA)
_
Microcrystalline cellulose Diluent 62
200 (MCC) _
---~---.
Magnesium stearate--_~_ Lubricant - 2.5
Purified water I Solvent 93
------------__._.___._~-- --- ---.__._.
Batch size 673 g
[00107] A flow chart detailing one method of preparing the tablet above is set
forth in Figure 8. BHA, polysorbate 80, and purified water were mixed to
create granulation fluid, which was stored overnight. Lactose anhydrous,
lactose fast flo, and the first portion of croscarmellose sodium were weighed
and screened through a coarse mesh, then combined with A-002 and
simvastatin (which can also be sieved if necessary) and dry blended for 1-2
minutes using a mixer at slow speed. In certain embodiments of this protocol,
112
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
microcrystalline cellulose may be included in the dry mix. Hydroxypropyl
cellulose was combined with the granulation fluid, resulting in greatly
increased viscosity. The resultant solution was added slowly to the mixer
containing the A-002/simvastatin mix. During addition, mixing speed was
gradually increased. Granulation end-point was reached when fine granules
formed without the mass becoming wet or sticky.
[00108] The granulation was screened through coarse mesh onto foil lined
trays and placed in an oven at 50 C and ambient humidity for approximately
three hours. After drying (and optional milling and/or sieving), granules were
placed in a low-density polyethylene (LDPE) bag. Approximately one-fourth
of the dried granules were pre-blended with microcrystalline cellulose and
magnesium stearate in a small baggy, and then the remainder of the granules
were combined with the pre-blend in a mixer. The second portion of
croscarmellose sodium was added in, and the entire mixture was mixed for
several minutes. Tablets were prepared by weighing approxirnately 580 mg
of the final blend into a tablet press and compressing at 200-2500 psi (Carver
press). The resultant tablets were weighed individually, and had an average
weight of approximately 0.6 g. Tablets appeared off-white to light tan in
color_
[00109] Two representative tablets (numbers 19 and 27) were analyzed by
reverse phase HPLC. Tablets were placed in a stability chamber at
25 C/60%RH. Representative HPLC results for two of the tablets are set forth
in Figure 9 and summarized in Table 29. Tablets maintained high potency
(i.e., high concentrations of A-002 and simvastatin), thereby validating the
feasibility of generating A-002/statin combination tablets for the treatment
of
various conditions.
113
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
Table 29: HPLC analysis of A-002/simvastatin combination tablets:
Tablet # Weight % A-002 % simvastatin
(RT: 28.5 minutes) (RT: 44.5 minutes)
19 0.64 g 92% 89%
27 0.60 g 88% 86% [00110] A film coating may be applied to combination A-
002/statin tablets
using methods well known in the art. In one such method, the coating
suspension is prepared by adding a film coat mixture such as Opadry YS-1-
18027-A to purified water in a mixing vessel. The mixing speed is reduced to
avoid foaming and the suspension is mixed for 60 or more minutes, until
uniform. After mixing, the suspension is allowed to stand for 60 or more
minutes to deaerate. A coater such as the Accela Coater is set up and the
theoretical amount of coating suspension to be sprayed is calculated based
on the number and size of tablets being coated. Tablets are loaded onto the
coating pan, and the nozzle air pressure (around 80-100 psi), atomizing air
pressure (around 25-45 psi), pattern air pressure (around 20-40 psi), delivery
rate (around 300-400 g/minute), pan speed (around 4-10 rpm), and dew point
setting (around 5-15 C) are verified. The fan is turned on, and gun-to-bed
distance (around 6-10 inches), supply air temperature (around 58 C), exhaust
air temperature (around 45 C), air volume inlet (around 1500 cfm), and
negative pan pressure differential are verified. The tablets are pre-heated,
and a sample number of tablets are weighed to determine the average core
tablet weight. The coating solution is sprayed onto the tablets while being
gently mixed. At various intervals of around 5-15 minutes, the average tablet
weight is recalculated. Once target tablet weight is reached, the supply air
temperature is reduced to around 45"C and the pan is jogged at intervals for
around five minutes to allow the tablets to dry. The supply air temperature is
114
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
reduced to around 30 C and tablets are jogged manually for around ten more
minutes. The coated tablets are then discharged from the coating pan.
Example 6: Once a day dosing of A-002 or A-002 plus statin:
[00111] 135 human subjects over the age of 18 with CVD, specifically stable
CAD, were randomized to receive either placebo or one of two dosages (250
mg or 500 mg) of A-002 via once a day oral administration over an eight week
time period. 89 of the subjects received A-002, while 46 of the subjects
received placebo. Subjects that were receiving statins or other compounds
used in the treatment of CVD at the outset of the trial continued to receive
those therapeutics throughout the trial. 121 of the 135 subjects were on
statins during the trial. There was little variation between dosage groups
with
respect to age, height, weight, or BMI.
[00112] Levels of various lipids and inflammatory markers were measured at
the outset of the trial and at weeks two, four, and/or eight weeks after the
start
of A-002 administration. In addition, plasma A-002 levels were measured at
various intervals_ Lipids that were measured included LDL, small LDL
particle, oxidized LDL, non-HDL cholesterol, total cholesterol, ApoB, and
triglycerides. In addition, levels of the inflammatory marker CRP were
rneasured. Lipid and inflammatory marker levels at each timepoint were
compared to the baseline measurements to determine the effect of A-002
administration. For data sets with a normal distribution, mean levels of
lipids
or inflammatory markers were analyzed. For data sets with non-normal
distribution, median levels were analyzed. LDL particle size was also
assessed.
115
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
[00113] Subjects in the ITT population receiving A-002 at either dosage
exhibited substantial decreases at week eight in rnean serum LDL, non-HDL
cholesterol, and total cholesterol levels (Tables 30-32, respectively) and in
median small LDL particle, oxidized LDL, TG, and ApoB levels (Tables 33-
36). Subjects receiving A-002 also exhibited an increase in mean LDL
particle size (Table 37). In addition, subjects receiving A-002 did not show
the
same increase in median CRP levels that was observed in subjects receiving
placebo (Table 38).
Table 30: Changes in serum LDL concentration in ITT population following A-
002 administration:
A-002 Placebo
Baseline # of 89 46
subjects _
Mean 75.4 mg/di 82.0 mg/dl
[LDL]
------- - ----- ----------- --------------..--
Week 8 # of 81 43
subjects
observed
Change in -7.1 mg/dl -0.9 mg/dl
mean
[LDL] vs.
baseline
% change -8.3% -0.7%
vs.
baseline
Table 31: Changes in serum non-HDL cholesterol concentration in ITT
population following A-002 administration:
__A-002 Placebo
Baseline # of 89 46
sub'ects
--f ---------- -----------.~ __ _ - -
Mean [non- 105.9 mg/dl 112.5 mg/dl
HDL
cholesterol]
------
Week 8 # of 81 43
subjects
observed
Change in -10.5 mg/dl -2.1 mg/dl
116
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
mean [non-
HDL
cholesterol]
vs. baseline
% change -9.9% -0.7%
vs. baseiine
Table 32: Changes in serum total cholesterol concentration in ITT population
following A-002 administration:
A-002 Piacebo
Baseline # of 89 46
subjects
Mean [total 154.9 mg/d) 159.9 mg/dl
cholesterol]
Week 8 # of ----- 81 -------- ---- 43 subjects
observed
Change in -11.5 mg/dl -3.1 mg/dl
mean [total
cholesterol]
vs. baseline
% change -7.3% -1.7%
vs. baseline
Table 33: Changes in serum small LDL particle concentration in ITT
population following A-002 administration:
A-002 Placebo
Baseline # of 89 46
subjects Median 759.0 nmol/L 782.0 nmol/L
[small
LDL
particle] Week 8 # of 81 43
subjects
observed
Change in -74.0 nmol/L +2.0 nmol/L
median
[small
LDL
particle]
vs.
baseline
/o change -9.1% +0.6%
vs.
baseline
117
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
Table 34: Changes in serum oxidized LDL concentration in ITT population
following A-002 administration:
___ -------- -----A-002 Placebo----
- ------ - - - ..-~
Baseline # of 89 46
sub'ects
Median 41.5 U/L 43.3 U/L
[oxidized
LDL]
Week 8# of 81 43
subjects
observed
--____.__----
Change in -1.4 U/L +0.3 U/L
median
[oxidized
LDL] vs.
baseline
% change -3.6% +0.4%
vs.
baseline
Table 35: Changes in serum TG concentration in ITT population following A-
002 administration: ( A-002 Placebo
Baseline # of 88 45
subea cts
Median 137-5 mg/di 135.0 mg/dl
[TG]
Week 8 # of 81 43
subjects
observed
Change in -13.0 mg/dl +3_0 mg/di
median
[TG] vs.
baseline
% change -9.8% +2.6%
vs.
baseline
Table 36: Changes in serum ApoB concentration in ITT population following
A-002 administration:
__------ _. - ~-. ~
A-002 Piacebo
----
Baseline # of 89 46
subects
_--___________._
Median 68.6 mg/dl ~ 62.8 mq/di
118
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577 --
Week 8 # of 81 43
subjects
observed
Change in -8.1 mg/di +2.5 mg/dl
median
[ApoB] vs.
baseline
% change -11.9% +3.5%
vs.
baseline
Table 37: Changes in LDL particle size in ITT population following A-002
administration:
-__-__-- ,----.---
A-002 Placebo
Baseline # of 89 46
subjects
Mean LDL 20.50 nm 20.50 nm
particle
size
Week 8 # of 81 43
subjects
observed
Change in +0.11 nm 0.00 nm
mean LDL
particle
size vs.
baseline
% change +0.6% 0.0%
vs.
baseline
Table 38: Changes in serum CRP concentration in ITT population following A-
002 administration:
A-002 Placebo
__..------- ------------ ----_____ _.--~ _.__ _._._
Baseline # of 89 46
subjects
Median 1.10 mg/L 1.30 mg/L
[CRP]
Week 8 # of - ---- 81------ ------ 43 subjects
observed
Change in +0.10 ng/ml +0.25 ng/mi
median
[CRP] vs.
baseline
119
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
% change +5.4% +25.4%
vs.
baseline
[00114] The decrease in mean serum LDL, non-HDL cholesterol, and total
cholesterol levels and median small LDL particle, oxidized LDL, TG, and
ApoB levels, the increase in mean LDL particle size, and the lower than
expected increase in median CRP levels observed in the ITT population were
also observed at eight weeks in the 121 subject subpopulation that was
receiving statins during the trial (Tables 39-47, respectively), indicating
that
administration of A-002 plus statin results in a greater therapeutic effect on
these markers than administration of statin alone. Likewise, a decrease in
LDL, non-HDL cholesterol, total cholesterol, small LDL particle, oxidized LDL,
and ApoB levels, an increase in LDL particle size, and a lower than expected
increase in CRP levels were observed in 51 subjects within the statin
subpopulation that had baseline LDL levels greater than 70 mg/dl (Tables 48-
55). The number of subjects in the non-statin subpopulation was too low to
allow for a detailed statistical analysis of the combined effect of A-002 and
statins versus the effect of either compound alone. However, once additional
non-statin controls are obtained, it is expected that a synergistic decrease
in
LDL levels similar to that observed following twice daily A-002 administration
(Example 4) will be observed in subjects receiving A-002 once a day.
Table 39: Changes in serum LDL concentration in statin subpopulation
following A-002 administration:
--------- A-002 Placebo
Baseline # of 81 40
subts
------- ----------------
Mean 70.5 mg/dl 75.5 mg/dl
[LDLJ -- -
Week 8 # of 75 37
120
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
------ ----- -- - ---- - I _ _
subjects
o_bserved
Change in -6.7 mg/dl -2.7 mg/dl
mean
[LDL] vs.
baseline
% change -8.2 /o -1.8%
vs.
baseline
Table 40: Changes in serum non-HDL cholesterol concentration in statin
subpopulation following A-002 administration:
A-002 Placebo
Baseline # of -------- ---- 81 40 subjects
Mean [non- 100.4 mg/di 104.8 mg/di
HDL
cholesterol]
Week 8 # of - ------ 75 ----------37
subjects
observed
Change in -10.1 mg/dl -3.2 mg/dl
mean [non-
HDL
cholesterol]
vs. baseline
- ----
% change -9.9% -1.3%
vs. baseline .1 _ - - - - -
Table 41: Changes in serum total cholesterol concentration in statin
subpopulation following A-002 administration:
-----A-002---- -_-__Placebo
Baseline # of 81 40
subjects
Mean [total 149.3 mg/dl 152.6 mg/dl
cholesterol]
Week 8 # of 75 37
subjects
observed
Change in -11.2 mg/di -4-4 mg/dl
mean [total
cholesterol]
vs. baseline
% change -7.3% -2.3%
vs. baseline
121
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
Table 42: Changes in serum small LDL particle concentration in statin
subpopulation following A-002 administration:
-------------------- --- A-002 ---- Placebo
Baseline # of 81 40
subjects Median 743.0 nmol/L 773.5 nmol/L
[small
LDL
particle]
Week 8 # of ----- 75 -------- -------37----_
subjects
observed
Change in -66.0 nmol/L -6.5 nmol/L
median
[small
LDL
particle]
vs.
baseline
% change -8.2% -1.0%
vs.
baseiine
Table 43: Changes in serum oxidized LDL concentration in statin
subpopulation following A-002 administration:
------
A 002 Placebo
Baseline # of 81 40
sub'ects
--- ------- -------- -
Median 41.1 U/L 42.2 U/L
[oxidized
LDL]
Week 8 # of 76 ----- ---- 33 subjects
observed
Change in -1.1 U/L +0.3 U/L
median
[oxidized
LDL] vs.
baseline
----- --- ---- ------ - ----
/a change -2.9% +0.4%
vs.
baseline
Table 44: Changes in serum TG concentration in statin subpopulation
following A-002 administration:
122
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
A-002 Placebo
Baseline # of ---~ ------- 80 39
sub'ects
~
Median 136.5 mg/dl 130.0 mg/dl
[TG]
Week 8 # of 75 37
subjects
observed
Change in -13.0 mg/dl +3.0 mg/dl
median
[TG] vs.
baseline
% change -10_9% +2.6%
vs.
baseline
Table 45: Changes in serum ApoB concentration in statin subpopulation
following A-002 administration:
A-002 Placebo
Baseline # of 81 40
sub'ects_
Median 66.1 mg/dl 58.2 mg/dl
[ApoB]
Week 8 # of 75 37
subjects
observed
Change in -7.9 mg/df +2.5 mg/dl
median
[ApoB] vs.
baseline
% change -11.9% +3.5%
vs.
baseline L
Table 46: Changes in LDL particle size in statin subpopulation following A-002
administration:
__---
A 002 Placebo
Baseline # of 81 40
subjects
- ---- - ------------
Mean LDL 20.50 nm 20.48 nm
particle
size
Week 8 # of 75 37
subjects
observed _ _ _ _
C h a n g in +0.10 nm -0.01 nm-
123
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
mean LDL
particle
size vs.
baseline
_-_
% change +0 5% --- --- 0.0%
vs.
baseline
Table 47: Changes in serum CRP concentration in statin subpopulation
following A-002 administration:
_ --__.-_-. ---------------
A-002 Placebo
Baseline # of 81 40
subjects Median 1.10 mg/L 1.30 mg/L
[CRP]
-_--- ----_, _ ___ _ _..------------ ...---- -_.
Week 8 # of 75 37
subjects
observed
Change in +0.10 mg/L +0.30 mg/L
median
[CRP] vs.
baseline
% change +8.7% +40.6%
vs.
baseline
Table 48: Changes in serum LDL concentration in statin subpopulation with
baseline serum LDL levels greater than 70 mg/dl following A-002
administration:
A-002 Placebo
Baseline # of 31 20
subjects
Mean 91.6 mg/dl 93.2 mg/d!
[LDL] Week 8 # of 30 18
subjects
observed
Change in -13.9 mg/dl -5.0 mg/dl
mean
[LDL] vs.
baseline
---- --- ------------ ------------ -
% change -15.0% -3.5%
vs.
baseline
124
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
Table 49: Changes in serum non-HDL cholesterol concentration in statin
subpopulation with baseline serum LDL levels greater than 70 mg/dl following
A-002 administration:
A-002 Placebo
Baseline # of 31 20
subjects Mean [non- 123.3 mg/dl 126.6 mg/dl
HDL
cholesterol]
Week 8# of 30 18
subjects
observed
Change in -16.7 mg/dl -7.2 mg/dl
mean [non-
HDL
cholesterol]
vs. baseline
% change -13.7 /o -4.1 %
vs. baseline
Table 50: Changes in serum total cholesterol concentration in statin
subpopulation with baseline serum LDL levels greater than 70 mg/di following
A-002 administration:
A-002 Placebo
Baseline # of 31 20
subjects
Mean [total 172.5 mg/dl 176.6 mg/dl
cholesterol]
Week 8 # of 30 18
subjects
observed
Change in -18.0 mg/dl -8.6 mg/d(
mean [total
cholesterol]
vs. baseline
% change -10.5% -4.2%
vs. baseline
Table 51: Changes in serum small LDL particle concentration in statin
subpopulation with baseline serum LDL levels greater than 70 mg/dl following
A-002 administration:
125
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
-------.-
A-002 Placebo
Baseline # of 31 20
sub'ects __ _ _ _
Median 964.0 nmol/L 919.5 nmol/L
[smal!
LDL
particle]
Week 8# of 30 18
subjects
observed
Change in -115.0 nmol/L -70.0 nmol/L
median
[small
LDL
particle]
vs.
baseline
% change -12.3% -7.6%
vs.
baseline
Table 52: Changes in serum oxidized LDL concentration in statin
subpopulation with baseline serum LDL levels greater than 70 mg/dl following
A-002 administration:
A-002 Placebo
Baseline # of 31 20
sub'ects
--------------
Median 47.9 U/L 51.6 U/L
[oxidized
LDL]
Week 8 # of 76 33
subjects
observed
Change in -2.8 U/L +1.3 U/L
median
[oxidized
LDL] vs.
baseline
% change -5.6% +2.5%
VS.
baseline
Table 53: Changes in serum ApoB concentration in statin subpopulation with
baseline serum LDL levels greater than 70 mg/d1 following A-002
administration:
126
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
__------
A-002 Placebo
Baseline # of 31 20
sub-ects
~____
Median 84.6 mg/dl 71.9 mg/dl
[ApoB]
Week 8 # of 30 18
subjects
observed
Change in -13.1 mg/dl +1.1 mg/di
median
[ApoB] vs.
baseline
% change -18.7% +2.1 %
vs.
baseline
Table 54: Changes in LDL particle size in statin subpopulation with baseline
serum LDL levels greater than 70 mg/di following A-002 administration:
_ A-002 Placebo
Baseline # of 31 20
subjects Mean LDL 20.24 nm 20.62 nm
particle
size
Week 8 # of 30 18
subjects
observed
- - - ---------
Change in +0.19 nm +0.02 nm
mean LDL
particle
size vs.
baseline
% change +1.0% +0.1%
vs.
baseline
Table 55: Changes in serum CRP concentration in statin subpopulation with
baseline serum LDL levels greater than 70 mg/di following A-002
administration:
-------
A-002 Placebo
Baseline # of 31 20
subjects
Median 1.10 mg/L 1.30 mg/L
[CRP] Week 8 # of 30 18 LL
127
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
subjects
observed
Change in +0.10 mg/L +0.45 mg/L
median
[CRP] vs.
baseline
lo change +9.1 % +26.0%
vs.
baseline
[00115] As stated above, the foregoing is merely intended to illustrate
various
embodiments of the present invention. The specific modifications discussed
above are not to be construed as limitations on the scope of the invention. It
will be apparent to one skilled in the art that various equivalents, changes,
and modifications may be made without departing from the scope of the
invention, and it is understood that such equivalent embodiments are to be
included herein. All references cited herein are incorporated by reference as
if fully set forth herein.
128
CA 02686157 2009-11-02
WO 2008/137803 PCT/US2008/062577
REFERENCES
1. Boekholdt et al. 2005. Arterioscler Thromb Vasc Biol 25:839-846.
2. Bostrom et al. 2007. Arterioscler Thromb Vasc Biol 27:600-606.
3. Camejo et al. 1998. Atherosclerosis 139:205-222.
4. Chait, A., et al. 2005. J Lipid Res 46:389-403.
5. Daugherty et al. 2000. J Clin Invest 105:1605-1612.
6. Elinder et al. 1997. Arterioscier Thromb Vasc Biol 17:2257-2263.
7, Hakala et al. 2001. Arterioscier Thromb Vasc Biol 21:1053-1058.
8. Hartford et al. 2006. J Cardiol 108:55-62.
9. lvandic et al. 1999. Arterioscler Thromb Vasc Biol 19:1284-1290_
10. Jialal, I. 1998. Clin Chem 44:1827-1832.
11. Kimura-Matsumoto et ai. Atherosclerosis Epub March 10, 2007.
12. Kugiyama et al. 1999. Circulation 100:1280-1284.
13. Liu et al. 2003. Eur Heart J 24:1824-1832.
14. Mallat et al. 2007. Arterioscler Thromb Vasc Biol 27:1177-1183.
15. Mallat et al. 2005. J Am Coll Cardiol 46:1249-1257.
16. Menschikowski et al, 1995. Atherosclerosis 118:173-181.
17. Nijmeijer et al. 2002. Cardiovasc Res 53:138-146.
18. Pruzanski et al. 1998. J Lipid Res 39:2150-2160.
19. Ramoner et al. 2005. Blood 105:3583.
20. Rosengren, B., et al. 2006. Arterioscler Thromb Vasc Biol 26:1579-
1585.
21. Sartipy et al. 1999. J Biol Chem 274:25913-25920.
22. Szmitko et al. 2003. Circulation 108:2041.
23. Tietge et al. 2000. J Biol Chem 275:10077-10084.
129
