U.S. patent application number 10/942438 was filed with the patent office on 2005-03-31 for treatment of insulin resistance syndrome and type 2 diabetes with pde9 inhibitors.
This patent application is currently assigned to Pfizer Inc. Invention is credited to Fryburg, David A., Gibbs, Earl Michael.
Application Number | 20050070557 10/942438 |
Document ID | / |
Family ID | 23318571 |
Filed Date | 2005-03-31 |
United States Patent
Application |
20050070557 |
Kind Code |
A1 |
Fryburg, David A. ; et
al. |
March 31, 2005 |
Treatment of insulin resistance syndrome and type 2 diabetes with
PDE9 inhibitors
Abstract
This invention is directed to a method of treating insulin
resistance syndrome (IRS), hypertension and/or type 2 diabetes in a
mammal comprising administering to said mammal a cGMP PDE9
inhibitor or a pharmaceutical composition thereof. This invention
is also directed to such methods wherein said cGMP PDE9 inhibitor
is used in combination with other agents to treat IRS, hypertension
and/or type 2 diabetes.
Inventors: |
Fryburg, David A.; (East
Lyme, CT) ; Gibbs, Earl Michael; (Oakdale,
CT) |
Correspondence
Address: |
PFIZER INC.
PATENT DEPARTMENT, MS8260-1611
EASTERN POINT ROAD
GROTON
CT
06340
US
|
Assignee: |
Pfizer Inc
|
Family ID: |
23318571 |
Appl. No.: |
10/942438 |
Filed: |
September 15, 2004 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10942438 |
Sep 15, 2004 |
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10283814 |
Oct 29, 2002 |
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60336981 |
Nov 2, 2001 |
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Current U.S.
Class: |
514/262.1 |
Current CPC
Class: |
A61P 3/04 20180101; A61P
3/10 20180101; A61P 9/10 20180101; A61P 3/08 20180101; A61K 45/06
20130101; A61P 7/02 20180101; A61P 5/50 20180101; A61P 43/00
20180101; A61P 3/06 20180101; A61K 31/505 20130101; A61P 19/06
20180101; A61K 2300/00 20130101; A61K 2300/00 20130101; A61K
2300/00 20130101; A61K 2300/00 20130101; A61K 31/5377 20130101;
A61P 9/12 20180101; A61K 31/541 20130101; A61K 31/437 20130101;
A61K 31/519 20130101; A61K 31/505 20130101; A61K 31/541 20130101;
A61K 31/519 20130101; A61K 31/5377 20130101 |
Class at
Publication: |
514/262.1 |
International
Class: |
A61K 031/519 |
Claims
1-4. (cancel).
5. A method of treating type 2 diabetes in a mammal comprising
administering to said mammal a cGMP PDE9 inhibitor, or salt or a
pharmaceutically acceptable salt thereof, or a solvate of said
inhibitor or said salt.
6. A method of claim 5 wherein said cGMP PDE9 inhibitor is a
compound of the formula (I) 212or a pharmaceutically acceptable
salt, thereof, or a solvate of said compound or said salt, wherein:
R.sup.1 is H or (C.sub.1-C.sub.6)alkyl; R.sup.2 is
(C.sub.1-C.sub.6)alkyl, straight chain or branched chain,
(C.sub.3-C.sub.7)cycloalkyl or heteroaryl; R.sup.3 is
(C.sub.1-C.sub.6)alkyl, straight chain or branched chain,
optionally substituted by 1-2 groups each independently selected
from Ar, (C.sub.3-C.sub.7)cycloalkyl, OAr, SAr,
NC(O)(C.sub.1-C.sub.6)alkyl, heteroaryl, xanthene, and naphthalene;
Ar is a group of formula 213wherein R.sup.4, R.sup.5 and R.sup.6
are each independently selected from H, halo, phenoxy, phenyl,
CF.sub.3, OCF.sub.3, S(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkyl, O(C.sub.1-C.sub.6)alkyl, said alkyl
optionally substituted by a heteroaryl group or by a phenyl group,
wherein said phenyl group is optionally substituted by 1-3 groups
selected from halo, CF.sub.3, OCF.sub.3 and (C.sub.1-C.sub.6)alkyl;
or wherein R.sup.4 and R.sup.5 may combine to form a
(C.sub.2-C.sub.3)alkyl link, wherein said link may optionally
incorporate a heteroatom selected from O, S and N; and heteroaryl
is aromatic 5-6 membered heterocycle containing 1-3 heteroatoms,
each independently selected from O, S and N, said heterocycle
optionally substituted by 1-3 substituents, each independently
selected from (C.sub.1-C.sub.6)alkyl, halo and phenyl, said phenyl
optionally substituted by 1-3 groups selected from halo and
(C.sub.1-C.sub.6)alkyl; with the proviso that when R.sup.1 is
--CH.sub.3, R.sup.2 cannot be --CH.sub.2CH.sub.2CH.sub.3.
7. A method of claim 6 wherein R.sup.1 is H or CH.sub.3; R.sup.2 is
(C.sub.3-C.sub.4)alkyl, cyclopentyl or pyridinyl; R.sup.3 is
(C.sub.1-C.sub.3)alkyl, optionally substituted by 1-2 groups
selected from Ar, (C.sub.3-C.sub.7)cycloalkyl and heteroaryl;
R.sup.4, R.sup.5 and R.sup.6 are each independently selected from
H, halo, phenoxy, phenyl, CF.sub.3, OCF.sub.3,
S(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkyl,
O(C.sub.1-C.sub.6)alkyl; said alkyl in the definition of R.sup.4,
R.sup.5 and R.sup.6 is optionally substituted by a heteroaryl group
or by a phenyl group optionally substituted by 1-3 groups selected
from halo, CF.sub.3, OCF.sub.3 and (C.sub.1-C.sub.6)alkyl; or
wherein R.sup.4 and R.sup.5 may combine to form a C.sub.2 alkyl
link, said link incorporating an O atom; and heteroaryl is an
aromatic 5-6 membered heterocycle containing at least 2 nitrogen
atoms, said heterocycle optionally substituted by 1-3 substituents,
each independently selected from (C.sub.1-C.sub.6) alkyl, halo and
phenyl, said phenyl in the definition of heterocycle optionally
substituted by 1-3 groups selected from halo and (C.sub.1-C.sub.6)
alkyl.
8. A method of claim 5 comprising administering to said mammal
5-(3-chloro-benzyl)-3-isopropyl-1,6-dihydro-pyrazolo[4,3-d]pyrimidin-7-on-
e, or a pharmaceutically acceptable salt thereof.
9. A method of treating dyslipidemia in a mammal comprising
administering to said mammal a cGMP PDE9 inhibitor, or a
pharmaceutically acceptable salt thereof, or a solvate of said
inhibitor or said salt.
10. A method of claim 9 wherein said cGMP PDE9 inhibitor is a
compound of the formula (I) 214or a pharmaceutically acceptable
salt, thereof, or a solvate of said compound or said salt, wherein:
R.sup.1 is H or (C.sub.1-C.sub.6)alkyl; R.sup.2 is
(C.sub.1-C.sub.6)alkyl, straight chain or branched chain,
(C.sub.3-C.sub.7)cycloalkyl or heteroaryl; R.sup.3 is
(C.sub.1-C.sub.6)alkyl, straight chain or branched chain,
optionally substituted by 1-2 groups each independently selected
from Ar, (C.sub.3-C.sub.7)cycloalkyl, OAr, SAr,
NC(O)(C.sub.1-C.sub.6)alkyl, heteroaryl, xanthene, and naphthalene;
Ar is a group of formula 215wherein R.sup.4, R.sup.5 and R.sup.6
are each independently selected from H, halo, phenoxy, phenyl,
CF.sub.3, OCF.sub.3, S(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkyl, O(C.sub.1-C.sub.6)alkyl, said alkyl
optionally substituted by a heteroaryl group or by a phenyl group,
wherein said phenyl group is optionally substituted by 1-3 groups
selected from halo, CF.sub.3, OCF.sub.3 and (C.sub.1-C.sub.6)alkyl;
or wherein R.sup.4 and R.sup.5 may combine to form a
(C.sub.2-C.sub.3)alkyl link, wherein said link may optionally
incorporate a heteroatom selected from O, S and N; and heteroaryl
is aromatic 5-6 membered heterocycle containing 1-3 heteroatoms,
each independently selected from O, S and N, said heterocycle
optionally substituted by 1-3 substituents, each independently
selected from (C.sub.1-C.sub.6)alkyl, halo and phenyl, said phenyl
optionally substituted by 1-3 groups selected from halo and
(C.sub.1-C.sub.6)alkyl; with the proviso that when R.sup.1 is
--CH.sub.3, R.sup.2 cannot be --CH.sub.2CH.sub.2CH.sub.3.
11. A method of claim 10 wherein R.sup.1 is H or CH.sub.3; R.sup.2
is (C.sub.3-C.sub.4)alkyl, cyclopentyl or pyridinyl; R.sup.3 is
(C.sub.1-C.sub.3)alkyl, optionally substituted by 1-2 groups
selected from Ar, (C.sub.3-C.sub.7)cycloalkyl and heteroaryl;
R.sup.4, R.sup.5 and R.sup.6 are each independently selected from
H, halo, phenoxy, phenyl, CF.sub.3, OCF.sub.3,
S(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkyl,
O(C.sub.1-C.sub.6)alkyl; said alkyl in the definition of R.sup.4,
R.sup.5 and R.sup.6 is optionally substituted by a heteroaryl group
or by a phenyl group optionally substituted by 1-3 groups selected
from halo, CF.sub.3, OCF.sub.3 and (C.sub.1-C.sub.6)alkyl; or
wherein R.sup.4 and R.sup.5 may combine to form a C.sub.2 alkyl
link, said link incorporating an O atom; and heteroaryl is an
aromatic 5-6 membered heterocycle containing at least 2 nitrogen
atoms, said heterocycle optionally substituted by 1-3 substituents,
each independently selected from (C.sub.1-C.sub.6) alkyl, halo and
phenyl, said phenyl in the definition of heterocycle optionally
substituted by 1-3 groups selected from halo and (C.sub.1-C.sub.6)
alkyl.
12. A method of claim 9 wherein said dyslipidemia is
hypertriglyceridemia.
13. A method of claim 9 comprising administering to said mammal
5-(3-chloro-benzyl)-3-isopropyl-1,6-dihydro-pyrazolo[4,3-d]pyrimidin-7-on-
e, or a pharmaceutically acceptable salt thereof.
14. A method of treating impaired glucose tolerance in a mammal
comprising administering to said mammal a cGMP PDE9 inhibitor, a or
a pharmaceutically acceptable salt thereof, or a solvate of said
inhibitor or said salt.
15. A method of claim 14 wherein said cGMP PDE9 inhibitor is a
compound of the formula (I) 216or a pharmaceutically acceptable
salt, thereof, or a solvate of said compound or said salt, wherein:
R.sup.1 is H or (C.sub.1-C.sub.6)alkyl; R.sup.2 is
(C.sub.1-C.sub.6)alkyl, straight chain or branched chain,
(C.sub.3-C.sub.7)cycloalkyl or heteroaryl; R.sup.3 is
(C.sub.1-C.sub.6)alkyl, straight chain or branched chain,
optionally substituted by 1-2 groups each independently selected
from Ar, (C.sub.3-C.sub.7)cycloalkyl, OAr, SAr,
NC(O)(C.sub.1-C.sub.6)alkyl, heteroaryl, xanthene, and naphthalene;
Ar is a group of formula 217wherein R.sup.4, R.sup.5 and R.sup.6
are each independently selected from H, halo, phenoxy, phenyl,
CF.sub.3, OCF.sub.3, S(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkyl, O(C.sub.1-C.sub.6)alkyl, said alkyl
optionally substituted by a heteroaryl group or by a phenyl group,
wherein said phenyl group is optionally substituted by 1-3 groups
selected from halo, CF.sub.3, OCF.sub.3 and (C.sub.1-C.sub.6)alkyl;
or wherein R.sup.4 and R.sup.5 may combine to form a
(C.sub.2-C.sub.3)alkyl link, wherein said link may optionally
incorporate a heteroatom selected from O, S and N; and heteroaryl
is aromatic 5-6 membered heterocycle containing 1-3 heteroatoms,
each independently selected from O, S and N, said heterocycle
optionally substituted by 1-3 substituents, each independently
selected from (C.sub.1-C.sub.6)alkyl, halo and phenyl, said phenyl
optionally substituted by 1-3 groups selected from halo and
(C.sub.1-C.sub.6)alkyl; with the proviso that when R.sup.1 is
--CH.sub.3, R.sup.2 cannot be --CH.sub.2CH.sub.2CH.sub.3.
16. A method of claim 15 wherein R.sup.1 is H or CH.sub.3; R.sup.2
is (C.sub.3-C.sub.4)alkyl, cyclopentyl or pyridinyl; R.sup.3 is
(C.sub.1-C.sub.3)alkyl, optionally substituted by 1-2 groups
selected from Ar, (C.sub.3-C.sub.7)cycloalkyl and heteroaryl;
R.sup.4, R.sup.5 and R.sup.6 are each independently selected from
H, halo, phenoxy, phenyl, CF.sub.3, OCF.sub.3,
S(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkyl,
O(C.sub.1-C.sub.6)alkyl; said alkyl in the definition of R.sup.4,
R.sup.5 and R.sup.6 is optionally substituted by a heteroaryl group
or by a phenyl group optionally substituted by 1-3 groups selected
from halo, CF.sub.3, OCF.sub.3 and (C.sub.1-C.sub.6)alkyl; or
wherein R.sup.4 and R.sup.5 may combine to form a C.sub.2 alkyl
link, said link incorporating an O atom; and heteroaryl is an
aromatic 5-6 membered heterocycle containing at least 2 nitrogen
atoms, said heterocycle optionally substituted by 1-3 substituents,
each independently selected from (C.sub.1-C.sub.6) alkyl, halo and
phenyl, said phenyl in the definition of heterocycle optionally
substituted by 1-3 groups selected from halo and (C.sub.1-C.sub.6)
alkyl.
17. A method of claim 14 comprising administering to said mammal
5-(3-chloro-benzyl)-3-isopropyl-1,6-dihydro-pyrazolo[4,3-d]pyrimidin-7-on-
e, or a pharmaceutically acceptable salt thereof or of said
prodrug.
18. A method of treating polycystic ovary syndrome in a mammal
comprising administering to said mammal a cGMP PDE9 inhibitor, a or
a pharmaceutically acceptable salt thereof, or a solvate of said
inhibitor or said salt.
19. A method of claim 18 wherein said cGMP PDE9 inhibitor is a
compound of the formula (I) 218or a pharmaceutically acceptable
salt, thereof, or a solvate of said compound or said salt, wherein:
R.sup.1 is H or (C.sub.1-C.sub.6)alkyl; R.sup.2 is
(C.sub.1-C.sub.6)alkyl, straight chain or branched chain,
(C.sub.3-C.sub.7)cycloalkyl or heteroaryl; R.sup.3 is
(C.sub.1-C.sub.6)alkyl, straight chain or branched chain,
optionally substituted by 1-2 groups each independently selected
from Ar, (C.sub.3-C.sub.7)cycloalkyl, OAr, SAr,
NC(O)(C.sub.1-C.sub.6)alkyl, heteroaryl, xanthene, and naphthalene;
Ar is a group of formula 219wherein R.sup.4, R.sup.5 and R.sup.6
are each independently selected from H, halo, phenoxy, phenyl,
CF.sub.3, OCF.sub.3, S(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkyl, O(C.sub.1-C.sub.6)alkyl, said alkyl
optionally substituted by a heteroaryl group or by a phenyl group,
wherein said phenyl group is optionally substituted by 1-3 groups
selected from halo, CF.sub.3, OCF.sub.3 and (C.sub.1-C.sub.6)alkyl;
or wherein R.sup.4 and R.sup.5 may combine to form a
(C.sub.2-C.sub.3)alkyl link, wherein said link may optionally
incorporate a heteroatom selected from O, S and N; and heteroaryl
is aromatic 5-6 membered heterocycle containing 1-3 heteroatoms,
each independently selected from O, S and N, said heterocycle
optionally substituted by 1-3 substituents, each independently
selected from (C.sub.1-C.sub.6)alkyl, halo and phenyl, said phenyl
optionally substituted by 1-3 groups selected from halo and
(C.sub.1-C.sub.6)alkyl; with the proviso that when R.sup.1 is
--CH.sub.3, R.sup.2 cannot be --CH.sub.2CH.sub.2CH.sub.3.
20. A method of claim 19 wherein R.sup.1 is H or CH.sub.3; R.sup.2
is (C.sub.3-C.sub.4)alkyl, cyclopentyl or pyridinyl; R.sup.3 is
(C.sub.1-C.sub.3)alkyl, optionally substituted by 1-2 groups
selected from Ar, (C.sub.3-C.sub.7)cycloalkyl and heteroaryl;
R.sup.4, R.sup.5 and R.sup.6 are each independently selected from
H, halo, phenoxy, phenyl, CF.sub.3, OCF.sub.3,
S(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkyl,
O(C.sub.1-C.sub.6)alkyl; said alkyl in the definition of R.sup.4,
R.sup.5 and R.sup.6 is optionally substituted by a heteroaryl group
or by a phenyl group optionally substituted by 1-3 groups selected
from halo, CF.sub.3, OCF.sub.3 and (C.sub.1-C.sub.6)alkyl; or
wherein R.sup.4 and R.sup.5 may combine to form a C.sub.2 alkyl
link, said link incorporating an O atom; and heteroaryl is an
aromatic 5-6 membered heterocycle containing at least 2 nitrogen
atoms, said heterocycle optionally substituted by 1-3 substituents,
each independently selected from (C.sub.1-C.sub.6) alkyl, halo and
phenyl, said phenyl in the definition of heterocycle optionally
substituted by 1-3 groups selected from halo and (C.sub.1-C.sub.6)
alkyl.
21. A method of claim 18 comprising administering to said mammal
5-(3-chloro-benzyl)-3-isopropyl-1,6-dihydro-pyrazolo[4,3-d]pyrimidin-7-on-
e, or a pharmaceutically acceptable salt thereof.
22. A combination comprising a cGMP PDE9 inhibitor and one or more
of a protein kinase inhibitor; an AMP-activated protein kinase; a
weight loss agent; insulin; a PPAR-.gamma. agonist; a PPAR-.gamma.
antagonist; a PPAR-.alpha. agonist; a dual
PPAR-.gamma./PPAR-.alpha. agonist; a sorbitol dehydrogenase
inhibitor; a glycogen phosphorylase inhibitor; a biguanide; an
HMG-CoA reductase inhibitor; an aldose reductase inhibitor; or a
PDE5 inhibitor.
23. A combination of claim 22 wherein said cGMP PDE9 inhibitor is a
compound of the formula (I) 220or a pharmaceutically acceptable
salt, thereof, or a solvate of said compound or said salt, wherein:
R.sup.1 is H or (C.sub.1-C.sub.6)alkyl; R.sup.2 is
(C.sub.1-C.sub.6)alkyl, straight chain or branched chain,
(C.sub.3-C.sub.7)cycloalkyl or heteroaryl; R.sup.3 is
(C.sub.1-C.sub.6)alkyl, straight chain or branched chain,
optionally substituted by 1-2 groups each independently selected
from Ar, (C.sub.3-C.sub.7)cycloalkyl, OAr, SAr,
NC(O)(C.sub.1-C.sub.6)alkyl, heteroaryl, xanthene, and naphthalene;
Ar is a group of formula 221wherein R.sup.4, R.sup.5 and R.sup.6
are each independently selected from H, halo, phenoxy, phenyl,
CF.sub.3, OCF.sub.3, S(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkyl, O(C.sub.1-C.sub.6)alkyl, said alkyl
optionally substituted by a heteroaryl group or by a phenyl group,
wherein said phenyl group is optionally substituted by 1-3 groups
selected from halo, CF.sub.3, OCF.sub.3 and (C.sub.1-C.sub.6)alkyl;
or wherein R.sup.4 and R.sup.5 may combine to form a
(C.sub.2-C.sub.3)alkyl link, wherein said link may optionally
incorporate a heteroatom selected from O, S and N; and heteroaryl
is aromatic 5-6 membered heterocycle containing 1-3 heteroatoms,
each independently selected from O, S and N, said heterocycle
optionally substituted by 1-3 substituents, each independently
selected from (C.sub.1-C.sub.6)alkyl, halo and phenyl, said phenyl
optionally substituted by 1-3 groups selected from halo and
(C.sub.1-C.sub.6)alkyl; with the proviso that when R.sup.1 is
--CH.sub.3, R.sup.2 cannot be --CH.sub.2CH.sub.2CH.sub.3.
24. A combination of claim 23 wherein R.sup.1 is H or CH.sub.3;
R.sup.2 is (C.sub.3-C.sub.4)alkyl, cyclopentyl or pyridinyl;
R.sup.3 is (C.sub.1-C.sub.3)alkyl, optionally substituted by 1-2
groups selected from Ar, (C.sub.3-C.sub.7)cycloalkyl and
heteroaryl; R.sup.4, R.sup.5 and R.sup.6 are each independently
selected from H, halo, phenoxy, phenyl, CF.sub.3, OCF.sub.3,
S(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6) alkyl,
O(C.sub.1-C.sub.6)alkyl; said alkyl in the definition of R.sup.4,
R.sup.5 and R.sup.6 is optionally substituted by a heteroaryl group
or by a phenyl group optionally substituted by 1-3 groups selected
from halo, CF.sub.3, OCF.sub.3 and (C.sub.1-C.sub.6)alkyl; or
wherein R.sup.4 and R.sup.5 may combine to form a C.sub.2 alkyl
link, said link incorporating an O atom; and heteroaryl is an
aromatic 5-6 membered heterocycle containing at least 2 nitrogen
atoms, said heterocycle optionally substituted by 1-3 substituents,
each independently selected from (C.sub.1-C.sub.6) alkyl, halo and
phenyl, said phenyl in the definition of heterocycle optionally
substituted by 1-3 groups selected from halo and (C.sub.1-C.sub.6)
alkyl.
25. A combination of claim 22 wherein said cGMP PDE9 inhibitor is
5-(3-chloro-benzyl)-3-isopropyl-1,6-dihydro-pyrazolo[4,3-d]pyrimidin-7-on-
e, or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable vehicle, carrier or diluent.
26. A kit comprising: a) a first unit dosage form comprising a cGMP
PDE9 inhibitor, a or a pharmaceutically acceptable salt thereof, or
a solvate of said inhibitor or said salt and a pharmaceutically
acceptable carrier, vehicle or diluent; b) a second unit dosage
form comprising: a protein kinase inhibitor; an AMP-activated
protein kinase; a weight loss agent; insulin; a PPAR-.gamma.
agonist; a PPAR-.gamma. antagonist; a PPAR-.alpha. agonist; a dual
PPAR-.gamma./PPAR-.alpha. agonist; a sorbitol dehydrogenase
inhibitor; a glycogen phosphorylase inhibitor; a biguanide; an
HMG-CoA reductase inhibitor; an aldose reductase inhibitor; or a
PDE5 inhibitor; or solvate of said protein kinase inhibitor,
AMP-activated protein, weight loss agent, insulin, PPAR-.gamma.
agonist, PPAR-.gamma. antagonist, PPAR-.alpha. agonist, dual
PPAR-.gamma./PPAR-.alpha. agonist, sorbitol dehydrogenase
inhibitor, glycogen phosphorylase inhibitor, biguanide, vastatin,
aldose reductase inhibitor or PDE5 inhibitor; or a pharmaceutically
acceptable salt thereof, or a solvate of said inhibitor or said
salt and a pharmaceutically acceptable carrier, vehicle or diluent;
and c) a container.
27. A kit comprising: a) a first unit dosage form comprising a cGMP
PDE9 inhibitor, or a pharmaceutically acceptable salt thereof, or a
solvate of said inhibitor or said salt and a pharmaceutically
acceptable carrier, vehicle or diluent; b) a second unit dosage
form comprising a cGMP PDE5 inhibitor, or a pharmaceutically
acceptable salt thereof, or a solvate of said inhibitor or said
salt and a pharmaceutically acceptable carrier, vehicle or diluent;
c) a third unit dosage form comprising a cGMP PDE11 inhibitor, or a
pharmaceutically acceptable salt thereof, or a solvate of said
inhibitor or said salt and a pharmaceutically acceptable carrier,
vehicle or diluent; and d) a container.
28. A method of treating type 2 diabetes in a mammal comprising
administering to said mammal a compound that increases
intracellular cGMP in said mammal.
29. A method of treating insulin resistance syndrome in a mammal
comprising administering to said mammal a compound that increases
intracellular cGMP in said mammal.
30. A method of treating polycystic ovary syndrome in a mammal
comprising administering to said mammal a compound that increases
intracellular cGMP in said mammal.
31. A method of treating dyslipidemia in a mammal comprising
administering to said mammal a compound that increases
intracellular cGMP in said mammal.
32. A method of treating impaired glucose tolerance in a mammal
comprising administering to said mammal a compound that increases
intracellular cGMP in said mammal.
Description
FIELD OF THE INVENTION
[0001] This invention relates to the use of cGMP PDE9 inhibitors
for the treatment of type 2 diabetes, hyperglycemia, dyslipidemia,
impaired glucose tolerance, type 1 diabetes and/or insulin
resistance syndrome (IRS). This invention also relates to
combinations comprising cGMP PDE9 inhibitors and other agents, said
combinations being useful in treating type 2 diabetes,
hyperglycemia, dyslipidemia, impaired glucose tolerance, type 1
diabetes and/or insulin resistance syndrome.
BACKGROUND OF THE INVENTION
[0002] IRS, as defined herein, means the concomitant existence in a
subject of two or more of: hyperinsulinemia, dyslipidemia,
hypertension, type 2 diabetes or impaired glucose tolerance,
hyperuricemia or gout, a pro-coagulant state, atherosclerosis
and/or truncal obesity. At the center of IRS, also known as
"Syndrome X" and "Metabolic Syndrome" in the biomedical literature,
is the common feature of tissue resistance to the action of
insulin. This impaired biological response to insulin is manifested
in the metabolic and vascular effects of insulin. Although there
are monogenic syndromes of insulin resistance (IR), in which a
definite gene has been identified as the cause of insulin
resistance (such as leprechaunism), these are relatively rare. By
contrast, the more common presentation of the IRS is associated
with obesity (particularly abdominal) and appears to be
polygenic.
[0003] The adaptive response to IR in individuals having IRS
produces compensatory hyperinsulinaemia. As subjects with IRS
become progressively insulin resistant, they manifest varying
degrees of change in clinical parameters, including blood pressure,
and/or increased levels of serum glucose, and/or cholesterol and/or
triglycerides, and/or uric acid, and/or factors that increase
coagulation. Once these clinical parameters have changed enough,
the patient with IRS may differentially manifest well-recognized
clinical conditions or diagnoses.
[0004] 2. These conditions include: type 2 diabetes, hypertension
(high blood pressure), hyperlipidemia or dyslipidemia, particularly
(but not limited to) hypertriglyceridemia, hyperuricemia or gout,
and hypercoagulability (defined as an abnormal, increased tendency
for clots to form, particularly inside blood vessels). These
clinical conditions are well-recognized risk factors for
cardiovascular (coronary artery and cerebrovascular) disease.
[0005] While it is difficult to estimate the prevalence of IRS in
the general populace due to both the diversity of the collective
risk factors associated with the syndrome and the likelihood that
many individuals affected by IRS go undetected because they may
exhibit no exterior symptoms and have no prior history of coronary
heart disease, it is postulated that at a minimum the patient
population at risk for the development of IRS includes individuals
with obesity, particularly truncal (abdominal) obesity. Obesity is
an extremely common problem in the industrialized world and is
associated with the clinical conditions mentioned above. Thus, it
is very likely that the prevalence of IRS is very high. Considering
this potential patient group alone forms an immense population
potentially at risk for the development of complications of IRS.
For example in the United States in 1994, 23% of the population
aged between 20 and 74 had hypertension, which accounted for 5
deaths per 100,000 population (1997). There will be an estimated
154,392,000 patients with diabetes world-wide in the year 2000. Of
these, 15,000,000 will be in the US and 934,000 in the UK. The
burden of disease for ischaemic heart disease for both sexes in the
WHO region estimated for 1998 was 51,948,000 with a mortality of
7,375,000, constituting 13.7% of total mortality and ranking the
highest in the mortality score. The burden of diabetes in both
sexes in the WHO region estimated for 1998 was 11,668,000. Thus
there exists a large medical need for an effective and safe oral
therapy for the treatment of IRS and prevention of the development
of IRS and its clinical consequences.
[0006] Resistance to the effects of insulin is also observed in the
diminished biological response of the endothelium to the vascular
effects of insulin. That is, insulin promotes relaxation of blood
vessels at least in part through the action of nitric oxide (NO).
Nitric oxide generated in the endothelium then stimulates cGMP
production in blood vessels and causes them to relax or dilate.
This opening of the blood vessel allows more blood to flow, which
is particularly important when more blood flow is needed to
critical organs, like the heart. It has been demonstrated that
there is a decreased release of NO from the endothelium of patients
with IR. This decreased release of NO is not only from insulin, but
also from other important vasodilators like acetylcholine. This
so-called "endothelial dysfunction" contributes to the risk factors
for cardiovascular disease which are associated with IRS. The
vascular effect of insulin contributes to the effect of insulin to
regulate metabolism, particularly, but not necessarily limited to,
glucose metabolism.
[0007] NO also has direct effects on glucose uptake by skeletal
muscle. That is, treatment with a NO-donor substance, such as
nitroprusside, or with an analogue of cGMP in vitro increases
glucose uptake (transport by GLUT4 glucose transporters). This
vasodilation-independent pathway is described in G. J. Etgen, D. A.
Fryburg and E. M. Gibbs in Diabetes, 46, 1997 pp. 1915-1919, which
is incorporated herein by reference. Taken together, NO and cGMP
have direct target tissue (skeletal muscle) and vascular actions
that influence, mediate, or mimic the action of insulin.
[0008] Further effects of impaired NO release by the endothelium
include: increases in vascular smooth muscle cell (VSMC) growth,
proliferation and migration which are key steps in atherosclerotic
plaque formation that can lead to stroke; an increase in platelet
aggregation and adhesiveness; an increase of lipid peroxidation and
an effect on the inhibition of cell adhesion molecule expression
including vascular cell adhesion molecule (VCAM-1), intracellular
adhesion molecule (ICAM), E-selectin. Impaired endothelial NO
release also impacts on the activity of inflammatory cytokines such
as tumour necrosis factor-.alpha. (TNF-.alpha.), and the production
of monocyte chemoattractant factor through decreased activity of
the transcriptional activator nuclear factor kappa B. These effects
on the platelet are also cGMP driven.
[0009] Finally, there are examples in which the treatment of
factors contributing to IRS (e.g., obesity) or the treatment of IRS
itself improves many of these clinical conditions which at first
glance appear to be unrelated. For example, dieting alone or
pharmacotherapeutic agents that induce weight loss will decrease
blood pressure, blood glucose and triglycerides. Agents that are
designed to improve insulin sensitivity can also favorably alter
blood pressure, lipids, and blood glucose.
[0010] Successful diagnosis and treatment of patients with IRS with
a PDE9 inhibitor will lead to clinically relevant improvements in
blood'pressure, and/or serum glucose and/or insulin and/or lipids
and/or uric acid, and/or procoagulant factors. This treatment can
occur alone or in combination with other therapeutics that improve
IRS. Improvement in these clinical conditions should reduce the
risk of the development of cardiovascular disease in these patients
as well as other complications of these individual disorders
(including, but not limited to diabetic neuropathy, nephropathy,
and retinopathy).
[0011] While IRS has many manifestations, an important underlying
mechanistic basis for the condition resides in a resistance to both
the vascular and metabolic effects of insulin. It is also
understood that the underlying pathology of vascular resistance in
insulin resistance syndrome, is a diminished amount of NO produced
by the endothelial cells in response to insulin. There is impaired
signaling of insulin for glucose uptake in insulin resistant
individuals.
[0012] Amplification of the cGMP signal, using cGMP PDE9 inhibitors
in patients with IRS enhances the insulin glucose uptake signal and
improves insulin action at key tissues. Enhancing insulin
sensitivity improves clinical parameters of IRS results, inter
alia, in:
[0013] 1. Blood glucose control: In patients with type 2 diabetes
or impaired glucose tolerance, an improvement in insulin
sensitivity results in a decrease in plasma glucose concentrations
(either fasting or after an oral glucose tolerance test or a meal).
In a related manner, as regulated by the patient's pathophysiology,
there will be an improvement in serum insulin concentrations in
either the fasting state or after a glucose load or meal. These
improvements in blood glucose control, should the subjects have
type 2 diabetes, manifest as improvements in measures of long-term
blood glucose control, such as, but not limited to, hemoglobin A1c
(glycosylated hemoglobin) or fructosamine.
[0014] 2. Blood pressure: It is believed that improvement in
insulin sensitivity yields improvements in both systolic and
diastolic blood pressure.
[0015] 3. Lipids: Improvement in insulin resistance yields
improvements in serum lipids, including, but not limited to, serum
cholesterol and triglycerides.
[0016] 4. Uric Acid: Improvement in insulin resistance yields
improvements in serum uric acid.
[0017] 5. Coagulation Factors: It is believed that improvement in
insulin resistance restores normal factors that worsen the
procoagulant state.
[0018] cGMP PDE 9 inhibitors prevent the effect of the
phosphodiesterase 9 enzyme that converts cGMP to inactive GMP thus
increasing the amount of accumulated cGMP. This accumulation
amplifies the vasodilatory, metabolic, and anti-atherogenic effects
of the available nitric oxide and insulin. This amplification
action mitigates the adverse effects associated with IRS and
improve one or more of the associated conditions.
[0019] Diabetes mellitus is characterized by metabolic defects in
production and utilization of carbohydrates, resulting in elevated
blood glucose or hyperglycemia due to the failure to maintain
appropriate blood sugar levels. Research in the treatment of
diabetes has centered on attempts to normalize fasting and
postprandial blood glucose levels. Current treatments include
administration of exogenous insulin, oral administration of drugs
and dietary therapies and exercise regimens.
[0020] Two major forms of diabetes mellitus are recognized. Type 1
diabetes, or insulin-dependent diabetes, is the result of an
absolute deficiency of insulin, the hormone which regulates
carbohydrate utilization. Type 2 diabetes, or non-insulin dependent
diabetes, often occurs with normal, or even elevated levels of
insulin and appears to be the result of the inability of tissues to
respond appropriately to insulin. Complications of type 2 diabetes
include retinopathy, nephropathy, neuropathy, and coronary heart
disease, and are believed to be triggered by excessive protein
glycation, which in turn results from excessive levels of
circulating glucose. Reduction in hyperglycemia by treatment with a
PDE9 inhibitor will lower the level of protein glycation and result
in a diminution in these diabetic complications.
[0021] Polycystic ovary syndrome (PCOS) also known as
Stein-Leventhal syndrome or functional ovarian hyperandrogenism, is
a complex endocrine disorder associated with a long-term lack of
ovulation (anovulation) and an excess of androgens (male sex
hormones, e.g., testosterone) circulating in the blood. The
disorder is characterized by the formation of cysts in the ovaries,
a process related to the failure of the ovary to release an egg
(ovum). In the majority of cases, the ovaries become enlarged. PCOS
afflicts up to 22% of women during their childbearing years,
although only 10% of these women develop symptoms. It is one of the
most frequent causes of infertility in women.
SUMMARY OF THE INVENTION
[0022] This invention is directed to a method of treating IRS in a
mammal comprising administering to said mammal a cGMP PDE9
inhibitor, a prodrug or solvate thereof or a pharmaceutically
acceptable salt of said PDE9 inhibitor, prodrug or solvate. In a
preferred embodiment of this invention, said method comprises
administering a pharmaceutical composition comprising a cGMP PDE9
inhibitor, a prodrug or solvate thereof or a pharmaceutically
acceptable salt of said PDE9 inhibitor, prodrug or solvate.
Preferably, said pharmaceutical composition additionally comprises
a pharmaceutically acceptable vehicle, diluent or carrier.
[0023] This invention is also directed to a method of treating type
2 diabetes in a mammal comprising administering to said mammal a
cGMP PDE9 inhibitor, a prodrug or solvate thereof or a
pharmaceutically acceptable salt of said PDE9 inhibitor, prodrug or
solvate. In a preferred embodiment of this invention, said method
comprises administering a pharmaceutical composition comprising a
cGMP PDE9 inhibitor, a prodrug or solvate thereof or a
pharmaceutically acceptable salt of said PDE9 inhibitor, prodrug or
solvate. Preferably, said pharmaceutical composition additionally
comprises a pharmaceutically acceptable vehicle, diluent or
carrier.
[0024] This invention is also directed to a method of treating type
1 diabetes in a mammal comprising administering to said mammal a
cGMP PDE9 inhibitor, a prodrug or solvate thereof or a
pharmaceutically acceptable salt of said cGMP PDE9 inhibitor,
prodrug or solvate. In a preferred embodiment of this invention,
said method comprises administering a pharmaceutical composition
comprising a cGMP PDE9 inhibitor, a prodrug or solvate thereof or a
pharmaceutically acceptable salt of said cGMP PDE9 inhibitor,
prodrug or solvate. Preferably, said pharmaceutical composition
additionally comprises a pharmaceutically acceptable vehicle,
diluent or carrier.
[0025] This invention is also directed to a method of treating
impaired glucose tolerance in a mammal comprising administering to
said mammal a cGMP PDE9 inhibitor, a prodrug or solvate thereof or
a pharmacetically acceptable salt of said cGMP PDE9 inhibitor,
prodrug or solvate. In a preferred embodiment of this invention,
said method comprises administering a pharmaceutical composition
comprising a cGMP PDE9 inhibitor, a prodrug or solvate thereof or a
pharmaceutically acceptable salt of said cGMP PDE9 inhibitor,
prodrug or solvate. Preferably, said pharmaceutical composition
additionally comprises a pharmaceutically acceptable vehicle,
diluent or carrier.
[0026] This invention is also directed to a method of treating
dyslipidemia such as, but not limited to, hypertriglyceridemia and
high LDL cholesterol, in a mammal comprising administering to said
mammal a cGMP PDE9 inhibitor, a prodrug or solvate thereof or a
pharmacetically acceptable salt of said cGMP PDE9 inhibitor,
prodrug or solvate. Dyslipidemia, where used herein, means an
alteration of the lipid profile in blood. In a preferred embodiment
of this invention, said method comprises administering a
pharmaceutical composition comprising a cGMP PDE9 inhibitor, a
prodrug or solvate thereof or a pharmaceutically acceptable salt of
said cGMP PDE9 inhibitor, prodrug or solvate. Preferably, said
pharmaceutical composition additionally comprises a
pharmaceutically acceptable vehicle, diluent or carrier.
[0027] This invention is also directed to a method of treating
polycystic ovary syndrome in a mammal comprising administering to
said mammal a cGMP PDE9 inhibitor, a prodrug or solvate thereof or
a pharmacetically acceptable salt of said cGMP PDE9 inhibitor,
prodrug or solvate. In a preferred embodiment of this invention,
said method comprises administering a pharmaceutical composition
comprising a cGMP PDE9 inhibitor, a prodrug or solvate thereof or a
pharmaceutically acceptable salt of said cGMP PDE9 inhibitor,
prodrug or solvate. Preferably, said pharmaceutical composition
additionally comprises a pharmaceutically acceptable vehicle,
diluent or carrier.
[0028] In a further embodiment, this invention is directed to a
first combination comprising two active ingredients selected from a
cGMP PDE9 inhibitor, a prodrug, solvate or salt thereof and one or
more, independently selected, protein kinase inhibitor, prodrug,
solvate or salt thereof; an AMP-activated protein kinase activator,
prodrug, solvate or salt thereof; a weight loss agent, prodrug,
solvate or salt thereof; insulin; a PPAR-.gamma. agonist, prodrug,
solvate or salt thereof; a PPAR-.gamma. antagonist, prodrug,
solvate or salt thereof, a PPAR-.alpha. agonist, prodrug, solvate
or salt thereof; a dual PPAR-.gamma./PPAR-.alph- a. agonist,
prodrug, solvate or salt thereof; a sorbitol dehydrogenase
inhibitor, prodrug, solvate or salt thereof; a glycogen
phosphorylase inhibitor, prodrug, solvate or salt thereof; a
biguanide such as metformin, prodrug, solvate or salt thereof; an
HMG-CoA reductase inhibitor, prodrug, solvate or salt thereof; an
aldose reductase inhibitor, prodrug, solvate or salt thereof; a
PDE5 inhibitor, prodrug, solvate or salt thereof; a PDE11
inhibitor, prodrug, solvate or salt thereof; or a CETP inhibitor,
prodrug, solvate or salt thereof. An especially preferred
combination is a combination of a cGMP PDE9 inhibitor, a prodrug,
solvate or salt thereof and a PDE5 inhibitor, a prodrug, solvate or
salt therof. In a further embodiment, the invention is directed to
a pharmaceutical composition comprising said first combination and
a pharmaceutically acceptable vehicle, carrier or diluent. In a
further embodiment, this invention is directed to methods of
treating insulin resistance in a mammal comprising administering to
said mammal said first combination or a pharmaceutical composition
comprising said first combination. In a still further embodiment,
this invention is directed to a method of treating type 2 diabetes
in a mammal comprising administering to said mammal said first
combination or a pharmaceutical composition comprising said first
combination.
[0029] In a still further embodiment, this invention is directed to
a second combination comprising three active ingredients selected
from a cGMP PDE9 inhibitor, a prodrug, solvate or salt thereof; a
cGMP PDE5 inhibitor, a prodrug, solvate or salt thereof; and a cGMP
PDE11 inhibitor, a prodrug, solvate or salt thereof. In a further
embodiment, the invention is directed to a pharmaceutical
composition comprising said second combination and a
pharmaceutically acceptable vehicle, carrier or diluent. In a
further embodiment, this invention is directed to methods of
treating insulin resistance in a mammal comprising administering to
said mammal said second combination or a pharmaceutical composition
comprising said second combination. In a still further embodiment,
this invention is directed to a method of treating type 2 diabetes
in a mammal comprising administering to said mammal said second
combination or a pharmaceutical composition comprising said second
combination.
[0030] This invention is also directed to a kit comprising:
[0031] a) a first unit dosage form comprising a cGMP PDE9
inhibitor, a prodrug or solvate thereof or a pharmaceutically
acceptable salt of said compound, prodrug or solvate and a
pharmaceutically acceptable carrier, vehicle or diluent;
[0032] b) a second unit dosage form comprising:
[0033] a protein kinase inhibitor;
[0034] an AMP-activated protein kinase;
[0035] a weight loss agent;
[0036] insulin;
[0037] a PPAR-.gamma. agonist;
[0038] a PPAR-.gamma. antagonist;
[0039] a PPAR-.alpha. agonist;
[0040] a dual PPAR-.gamma./PPAR-.alpha. agonist;
[0041] a sorbitol dehydrogenase inhibitor;
[0042] a glycogen phosphorylase inhibitor;
[0043] a biguanide such as metformin
[0044] an HMG-CoA reductase inhibitor;
[0045] an aldose reductase inhibitor;
[0046] a PDE5 inhibitor;
[0047] a PDE11 inhibitor; or
[0048] a CETP inhibitor;
[0049] a prodrug or solvate of said protein kinase inhibitor,
AMP-activated protein, weight loss agent, insulin, PPAR-.gamma.
agonist, PPAR-.alpha. agonist, PPAR-.alpha. antagonist, dual
PPAR-.gamma./PPAR-.alpha. agonist, sorbitol dehydrogenase
inhibitor, glycogen phosphorylase inhibitor, biguanide, vastatin,
aldose reductase inhibitor, PDE5 inhibitor, PDE11 inhibitor or CETP
inhibitor; or a pharmaceutically acceptable salt thereof or of said
prodrug or solvate and a pharmaceutically acceptable carrier,
vehicle or diluent; and
[0050] c) a container.
[0051] This invention is also directed to a kit comprising:
[0052] a) a first unit dosage form comprising a cGMP PDE9
inhibitor, a prodrug or solvate thereof or a pharmaceutically
acceptable salt of said compound, prodrug or solvate and a
pharmaceutically acceptable carrier, vehicle or diluent;
[0053] b) a second unit dosage form comprising a cGMP PDE5
inhibitor, a prodrug or solvate thereof or a pharmaceutically
acceptable salt of said compound, prodrug or solvate and a
pharmaceutically acceptable carrier, vehicle or diluent;
[0054] c) a third unit dosage form comprising a cGMP PDE11
inhibitor, a prodrug or solvate thereof or a pharmaceutically
acceptable salt of said compound, prodrug or solvate and a
pharmaceutically acceptable carrier, vehicle or diluent; and
[0055] d) a container.
[0056] A further aspect of the invention provides for a method for
treating IRS as defined above in a polygenic insulin resistant
mammal comprising administering to the mammal an effective amount
of a cGMP PDE9 inhibitor or a pharmaceutical composition thereof.
It is a further aspect of this invention to treat such polygenic
insulin resistant mammals with a combination of a cGMP PDE9
inhibitor and a second compound as defined above, or with a
pharmaceutical composition comprising such a combination and a
pharmaceutically acceptable vehicle, carrier or diluent. In yet
another aspect of this invention, such polygenic insulin resistant
mammals are treated with a kit as described above.
[0057] The suitability of any particular cGMP PDE9 inhibitor can be
readily determined by evaluation of its potency and selectivity
using literature methods followed by evaluation of its toxicity,
absorption, metabolism, pharmacokinetics, etc in accordance with
standard pharmaceutical practice.
[0058] Preferably, the cGMP PDE9 inhibitors have an IC.sub.50 at
less than 100 nanomolar, more preferably, at less than 50
nanomolar, more preferably still at less than 15 nanomolar.
[0059] IC50 values for the cGMP PDE9 inhibitors may be determined
using the PDE9 assay in the Test Methods Section hereinafter.
[0060] It is to be understood that the contents of the above
published patent applications, and in particular the general
formulae and exemplified compounds therein are incorporated herein
in their entirety by reference thereto.
[0061] A preferred group of cGMP PDE9 inhibitors for use in the
methods, compositions, combinations and kits of the instant
invention include compounds of the formula (I) 1
[0062] or a pharmaceutically acceptable salt, solvate or prodrug
thereof,
[0063] wherein:
[0064] R.sup.1 is H or (C.sub.1-C.sub.6)alkyl;
[0065] R.sup.2 is (C.sub.1-C.sub.6)alkyl, straight chain or
branched chain, (C.sub.3-C.sub.7)cycloalkyl or heteroaryl;
[0066] R.sup.3 is (C.sub.1-C.sub.6)alkyl, straight chain or
branched chain, optionally substituted by 1-2 groups each
independently selected from Ar, (C.sub.3-C.sub.7)cycloalkyl, OAr,
SAr, NC(O)(C.sub.1-C.sub.6)alk- yl, heteroaryl, xanthene, and
naphthalene; Ar is a group of formula 2
[0067] wherein R.sup.4, R.sup.5 and R.sup.6 are each independently
selected from H, halo, phenoxy, phenyl, CF.sub.3, OCF.sub.3,
S(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkyl,
O(C.sub.1-C.sub.6)alkyl, said alkyl optionally substituted by a
heteroaryl group or by a phenyl group, wherein said phenyl group is
optionally substituted by 1-3 groups selected from halo, CF.sub.3,
OCF.sub.3 and (C.sub.1-C.sub.6)alkyl; or wherein R.sup.4 and
R.sup.5 may combine to form a (C.sub.2-C.sub.3)alkyl link, wherein
said link may optionally incorporate a heteroatom selected from O,
S and N; and
[0068] heteroaryl is aromatic 5-6 membered heterocycle containing
1-3 heteroatoms, each independently selected from O, S and N, said
heterocycle optionally substituted by 1-3 substituents, each
independently selected from (C.sub.1-C.sub.6)alkyl, halo and
phenyl, said phenyl optionally substituted by 1-3 groups selected
from halo and (C.sub.1-C.sub.6)alkyl;
[0069] with the proviso that when R.sup.1 is --CH.sub.3, R.sup.2
cannot be --CH.sub.2CH.sub.2CH.sub.3.
[0070] A particularly preferred group of compounds within the
preferred group are those compounds wherein R.sup.1 is H or
CH.sub.3. More preferably R.sup.1 is H.
[0071] Another particularly preferred group of compounds within the
preferred group are those compounds wherein R.sup.2 is selected
from (C.sub.3-C.sub.4)alkyl, cyclopentyl and pyridinyl. More
preferably R.sup.2 is 3-pyridinyl.
[0072] Another particularly preferred group of compounds within the
preferred group are those compounds wherein R.sup.3 is
(C.sub.1-C.sub.3)alkyl, optionally substituted by 1-2 groups
selected from: Ar, (C.sub.3-C.sub.7)cycloalkyl and heteroaryl. More
preferably R.sup.3 is (C.sub.1-C.sub.3)alkyl, optionally
substituted by Ar. Most preferably R.sup.3 is C, alkyl substituted
by Ar, where R.sup.4, R.sup.5 and R.sup.6 are each H.
[0073] Another particularly preferred group of compounds within the
preferred group are those compounds wherein R.sup.4, R.sup.5 and
R.sup.6 are each independently selected from: H, halo, phenoxy,
phenyl, CF.sub.3, OCF.sub.3, S(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6) alkyl, O(C.sub.1-C.sub.6)alkyl, said alkyl
optionally substituted by a heteroaryl group or by a phenyl group,
wherein said phenyl group is optionally substituted by 1-3 groups
selected from halo, CF.sub.3, OCF.sub.3 and (C.sub.1-C.sub.6)alkyl;
or wherein R.sup.4 and R.sup.5 may combine to form a C.sub.2 alkyl
link, said link incorporating an O atom. More preferably R.sup.4,
R.sup.5 and R.sup.6 are each independently selected from H, halo,
OCF.sub.3, CF.sub.3, OAr, and O(C.sub.1-C.sub.6)alkyl optionally
substituted by phenyl, optionally substituted by H, halo, phenoxy,
phenyl, CF.sub.3, OCF.sub.3, S(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6) alkyl, O(C.sub.1-C.sub.6) alkyl, said alkyl
optionally substituted by a heteroaryl group or by a phenyl group,
wherein said phenyl group is optionally substituted by H, halo,
CF.sub.3, OCF.sub.3 and (C.sub.1-C.sub.6) alkyl. Yet more
preferably R.sup.4, R.sup.5 and R.sup.6 are each independently
selected from Cl, H, OCF.sub.3, CF.sub.3 and
O(C.sub.1-C.sub.6)alkyl substituted by phenyl. Most preferably,
R.sup.4, R.sup.5 and R.sup.6 are each independently selected from
H, Cl and O(C.sub.1-C.sub.3)alkyl substituted by phenyl.
[0074] Another particularly preferred group of compounds within the
preferred group are those compounds wherein heteroaryl is an
aromatic 5-6 membered heterocycle containing at least 2 nitrogen
atoms, said heterocycle optionally substituted by 1-3 substituents,
each independently selected from (C.sub.1-C.sub.6) alkyl, halo and
phenyl, said phenyl optionally substituted by 1-3 groups selected
from halo and (C.sub.1-C.sub.6) alkyl. More preferably heteroaryl
is an aromatic 5 membered heterocycle containing at least 2
nitrogen atoms, said heterocycle optionally substituted by 1
substituent, each independently selected from (C.sub.1-C.sub.6)
alkyl, halo and phenyl, said phenyl optionally substituted by 1-3
groups selected from halo and (C.sub.1-C.sub.6) alkyl. Yet more
preferably heteroaryl is an aromatic 5 membered heterocycle
containing at least 2 nitrogen atoms, said heterocycle optionally
substituted by phenyl optionally substituted by halo. Most
preferably heteroaryl is an imidazole or an oxadiazole.
[0075] An especially preferred cGMP PDE9 inhibitor is
5-(3-chloro-benzyl)-3-isopropyl-1,6-dihydro-pyrazolo[4,3-d]pyrimidin-7-on-
e, a prodrug thereof or a pharmaceutically acceptable salt thereof
or of said prodrug.
[0076] According to a further aspect the present invention
additionally provides for the use of a PDE9 inhibitor or a
pharmaceutical composition thereof for the treatment of the insulin
resistance syndrome in a subject having type 2 diabetes mellitus or
impaired glucose tolerance or having a family history of diabetes
and at least one of the following conditions: dyslipidemia,
hypertension, hyperuricemia, a pro-coagulant state, atherosclerosis
or truncal obesity.
[0077] According to a further aspect the present invention
additionally provides a method of elevating intracellular cGMP in a
mammal in need thereof comprising administering to said mammal a
PDE9 inhibitor, a prodrug thereof, a pharmaceutically acceptable
salt of said PDE9 inhibitor or of said prodrug, or a pharmaceutical
composition comprising a PDE9 inhibitor. It is particularly
preferred that type 2 diabetes, insulin resistance syndrome or
hypertension is treated thereby.
[0078] According to a further aspect the invention additionally
provides a method of treating hypertension in a mammal comprising
administering to said mammal a PDE9 inhibitor, a prodrug thereof or
a pharmaceutically acceptable salt of said PDE9 inhibitor or of
said prodrug. It is particularly preferred that said PDE9 inhibitor
is
5-(3-chloro-benzyl)-3-isopropyl-1,6-dihydro-pyrazolo[4,3-d]pyrimidin-7-on-
e, a prodrug thereof or a pharmaceutically acceptable salt thereof
or of said prodrug.
DETAILED DESCRIPTION OF THE INVENTION
[0079] The PDE9 inhibitors used in the pharmaceutical compositions
and methods of this invention may be prepared as set forth in the
Examples provided below or by following procedures analogous to
those set forth in U.S. Pat. No. 6,235,742 B1, which is
incorporated herein by reference.
[0080] The pharmaceutically acceptable salts of the cGMP PDE9
inhibitor compounds as disclosed herein for use in the treatment of
the insulin resistance syndrome in accordance with the present
invention which contain a basic centre are, for example, non-toxic
acid addition salts formed with inorganic acids such as
hydrochloric, hydrobromic, hydroiodic, sulphuric and phosphoric
acid, with carboxylic acids or with organo-sulphonic acids.
Examples include the HCl, HBr, HI, sulphate or bisulphate, nitrate,
phosphate or hydrogen phosphate, acetate, benzoate, succinate,
saccarate, fumarate, maleate, lactate, citrate, tartrate,
gluconate, camsylate, methanesulphonate, ethanesulphonate,
benzene-sulphonate, p-toluenesulphonate and pamoate salts. The cGMP
PDE9 inhibitor compounds for use in the present invention can also
provide pharmaceutically acceptable metal salts, in particular
non-toxic alkali and alkaline earth metal salts, with bases.
Examples include the sodium, potassium, aluminium, calcium,
magnesium, zinc and diethanolamine salts. For a review on suitable
pharmaceutical salts see Berge et al, J. Pharm, Sci.,
66,1-19,1977.
[0081] The cGMP PDE9 inhibitor compounds suitable for use in
accordance with the present invention, their pharmaceutically
acceptable salts, and pharmaceutically acceptable solvates of
either entity can be administered alone but, in human therapy will
generally be administered in admixture with a suitable
pharmaceutical excipient diluent or carrier selected with regard to
the intended route of administration and standard pharmaceutical
practice.
[0082] For example, the cGMP PDE9 inhibitor compounds suitable for
use in accordance with the present invention or salts or solvates
thereof can be administered orally, buccally or sublingually in the
form of tablets, capsules (including soft gel capsules),
multi-particulate, gels, films, ovules, elixirs, solutions or
suspensions, which may contain flavouring or colouring agents, for
immediate-, delayed-, modified-, sustained-, dual-,
controlled-release or pulsatile delivery applications. Such
compounds may also be administered via fast dispersing or fast
dissolving dosages forms or in the form of a high energy dispersion
or as coated particles. Suitable pharmaceutical formulations may be
in coated or un-coated form as desired.
[0083] Such solid pharmaceutical compositions, for example, tablets
may contain excipients such as microcrystalline cellulose, lactose,
sodium citrate, calcium carbonate, dibasic calcium phosphate,
glycine and starch (preferably corn, potato or tapioca starch),
disintegrants such as sodium starch glycollate, croscarmellose
sodium and certain complex silicates, and granulation binders such
as polyvinylpyrrolidone, hydroxypropylmethyl cellulose (HPMC),
hydroxypropylcellulose (HPC), hydroxypropyl methylcellulose acetate
succinate (HPMCAS), sucrose, gelatin and acacia. Additionally,
lubricating agents such as magnesium stearate, stearic acid,
glyceryl behenate and talc may be included.
[0084] Solid compositions of a similar type may also be employed as
fillers in gelatin capsules or HPMC capsules. Preferred excipients
in this regard include lactose, starch, a cellulose, milk sugar or
high molecular weight polyethylene glycols. For aqueous suspensions
and/or elixirs, the cGMP PDE9 inhibitor compounds may be combined
with various sweetening or flavouring agents, colouring matter or
dyes, with emulsifying and/or suspending agents and with diluents
such as water, ethanol, propylene glycol and glycerin, and
combinations thereof.
[0085] Modified release and pulsatile release dosage forms may
contain excipients such as those detailed for immediate release
dosage forms together with additional excipients that act as
release rate modifiers, these being coated on and/or included in
the body of the device. Release rate modifiers include, but are not
exclusively limited to, HPMC, HPMCAS, methyl cellulose, sodium
carboxymethylcellulose, ethyl cellulose, cellulose acetate,
polyethylene oxide, Xanthan gum, Carbomer, ammonio methacrylate
copolymer, hydrogenated castor oil, carnauba wax, paraffin wax,
cellulose acetate phthalate, hydroxypropylmethyl cellulose
phthalate, methacrylic acid copolymer and mixtures thereof.
Modified release and pulsatile release dosage forms may contain one
or a combination of release rate modifying excipients. Release rate
modifying excipients maybe present both within the dosage form i.e.
within the matrix, and/or on the dosage form, i.e., upon the
surface or coating.
[0086] Fast dispersing or dissolving dosage formulations (FDDFs)
may contain the following ingredients: aspartame, acesulfame
potassium, citric acid, croscarmellose sodium, crospovidone,
diascorbic acid, ethyl acrylate, ethyl cellulose, gelatin,
hydroxypropylmethyl cellulose, magnesium stearate, mannitol, methyl
methacrylate, mint flavouring, polyethylene glycol, fumed silica,
silicon dioxide, sodium starch glycolate, sodium stearyl fumarate,
sorbitol, xylitol. The terms dispersing or dissolving as used
herein to describe FDDFs are dependent upon the solubility of the
drug substance used i.e. where the drug substance is insoluble a
fast dispersing dosage form can be prepared and where the drug
substance is soluble a fast dissolving dosage form can be
prepared.
[0087] The cGMP PDE9 inhibitor compounds suitable for use in
accordance with the present invention can also be administered
parenterally, for example, intracavernosally, intravenously,
intra-arterially, intraperitoneally, intrathecally,
intraventricularly, intraurethrally, intrasternally,
intracranially, intramuscularly or subcutaneously, or they may be
administered by infusion or needle-free techniques. For such
parenteral administration they are best used in the form of a
sterile aqueous solution which may contain other substances, for
example, enough salts or glucose to make the solution isotonic with
blood. The aqueous solutions should be suitably buffered
(preferably to a pH of from 3 to 9), if necessary. The preparation
of suitable parenteral formulations under sterile conditions is
readily accomplished by standard pharmaceutical techniques
well-known to those skilled in the art.
[0088] For oral and parenteral administration to human patients,
the daily dosage level of the cGMP PDE9 inhibitor compounds for use
in the present invention or salts or solvates thereof will usually
be from 1 to 500 mg (in single or divided doses). A preferred
dosage range is about 1 mg to about 100 mg. For the treatment of
IRS the dosage may by via single dose, divided daily dose, multiple
daily dose, continuous (chronic) daily dosing for a specified
period which may be from one to five or 5 or more, such as up to 10
or more days. Alternatively the treatment of IRS may be affected by
continuous dosing, such as for example, via a controlled release
dosage form wherein such continuous dosage form can be administered
on a daily basis for a number of days or wherein such continuous
dosing can be affected via a slow-release formulation which doses
for more than one day at a time.
[0089] Thus, for example, tablets or capsules of the cGMP PDE9
inhibitor compounds suitable for use in accordance with the present
invention or salts or solvates thereof may contain from 1 mg to 250
mg of active compound for administration singly or two or more at a
time, as appropriate. Preferred tablets or capsules will contain
about 1 mg to about 50 mg of active compound for administration
singly or two or more at a time, as appropriate. The physician in
any event will determine the actual dosage which will be most
suitable for any individual patient and it will vary with the age,
weight and response of the particular patient. The above dosages
are exemplary of the average case. There can, of course, be
individual instances where higher or lower dosage ranges are
merited and such are within the scope of this invention.
[0090] The cGMP PDE9 inhibitor compounds suitable for use in
accordance with the present invention can also be administered
intranasally or by inhalation and are conveniently delivered in the
form of a dry powder inhaler or an aerosol spray presentation from
a pressurised container, pump, spray or nebuliser with the use of a
suitable propellant, e.g. dichlorodifluoromethane,
trichlorofluoromethane, dichlorotetrafluoroethan- e, a
hydrofluoroalkane such as 1,1,1,2-tetrafluoroethane (HFA 134A
[trade mark] or 1,1,1,2,3,3,3-heptafluoropropane (HFA 227EA [trade
mark]), carbon dioxide or other suitable gas. In the case of a
pressurised aerosol, the dosage unit may be determined by providing
a valve to deliver a metered amount. The pressurised container,
pump, spray or nebuliser may contain a solution or suspension of
the active compound, e.g. using a mixture of ethanol and the
propellant as the solvent, which may additionally contain a
lubricant, e.g. sorbitan trioleate. Capsules and cartridges (made,
for example, from gelatin) for use in an inhaler or insufflator may
be formulated to contain a powder mix of a compound of the
invention and a suitable powder base such as lactose or starch.
[0091] Aerosol or dry powder formulations are preferably arranged
so that each metered dose or "puff" contains from 1 to 50 mg of a
compound of the invention for delivery to the patient. The overall
daily dose with an aerosol will be in the range of from 1 to 50 mg
which may be administered in a single dose or, more usually, in
divided doses throughout the day.
[0092] The cGMP PDE9 inhibitor compounds suitable for use in
accordance with the present invention may also be formulated for
delivery via an atomiser. Formulations for atomiser devices may
contain the following ingredients as solubilisers, emulsifiers or
suspending agents: water, ethanol, glycerol, propylene glycol, low
molecular weight polyethylene glycols, sodium chloride,
fluorocarbons, polyethylene glycol ethers, sorbitan trioleate,
oleic acid.
[0093] Alternatively, the cGMP PDE9 inhibitor compounds suitable
for use in accordance with the present invention or salts or
solvates thereof can be administered in the form of a suppository
or pessary, or they may be applied topically in the form of a gel,
hydrogel, lotion, solution, cream, ointment or dusting powder. The
cGMP PDE9 inhibitor compounds suitable for use in accordance with
the present invention or salts or solvates thereof may also be
dermally or transdermally administered, for example, by the use of
a skin patch. They may also be administered by the pulmonary or
rectal routes.
[0094] The compounds may also be administered by the ocular route.
For ophthalmic use, the compounds can be formulated as micronised
suspensions in isotonic, pH adjusted, sterile saline, or,
preferably, as solutions in isotonic, pH adjusted, sterile saline,
optionally in combination with a preservative such as a
benzylalkonium chloride. Alternatively, they may be formulated in
an ointment such as petrolatum.
[0095] For application topically to the skin, the cGMP PDE9
inhibitor compounds suitable for use in accordance with the present
invention or salts or solvates thereof can be formulated as a
suitable ointment containing the active compound suspended or
dissolved in, for example, a mixture with one or more of the
following: mineral oil, liquid petrolatum, white petrolatum,
propylene glycol, polyoxyethylene polyoxypropylene compound,
emulsifying wax and water. Alternatively, they can be formulated as
a suitable lotion or cream, suspended or dissolved in, for example,
a mixture of one or more of the following: mineral oil, sorbitan
monostearate, a polyethylene glycol, liquid paraffin, polysorbate
60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl
alcohol and water.
[0096] The cGMP PDE9 inhibitor compounds suitable for use in
accordance with the present invention may also be used in
combination with a cyclodextrin. Cyclodextrins are known to form
inclusion and non-inclusion complexes with drug molecules.
Formation of a drug-cyclodextrin complex may modify the solubility,
dissolution rate, bioavailability and/or stability property of a
drug molecule. Drug-cyclodextrin complexes are generally useful for
most dosage forms and administration routes. As an alternative to
direct complexation with the drug the cyclodextrin may be used as
an auxiliary additive, e.g. as a carrier, diluent or solubiliser.
Alpha-, beta- and gamma-cyclodextrins are most commonly used and
suitable examples are described in International Patent Application
Publication Nos. WO91/11172, WO94/02518 and WO98/55148.
[0097] Generally, in humans, oral administration is the preferred
route, being the most convenient. In circumstances where the
recipient suffers from a swallowing disorder or from impairment of
drug absorption after oral administration, the drug may be
administered parenterally, sublingually or buccally.
[0098] For veterinary use, a compound, or a veterinarily acceptable
salt thereof, or a veterinarily acceptable solvate or pro-drug
thereof, is administered as a suitably acceptable formulation in
accordance with normal veterinary practice and the veterinary
surgeon will determine the dosing regimen and route of
administration which will be most appropriate for a particular
animal.
[0099] It is to be appreciated that all references herein to
treatment include curative, palliative and prophylactic
treatment.
[0100] The present invention additionally comprises the use of a
combination of a first and a second compound for the treatment of
the insulin resistance syndrome or type 2 diabetes. The first
compound of the combination is a cGMP PDE9 inhibitor compound as
defined herein. The second compound of the combination is a
naturally occurring or synthetic prostaglandin or ester thereof; an
.alpha.-adrenergic receptor antagonist compound (also known as
.alpha.-adrenoreceptors, .alpha.-receptors or .alpha.-blockers); a
nitric oxide donor (also known as NO-donor or NO-agonist); a
potassium channel opener or potassium channel modulator; a
dopaminergic agent; a vasodilator agent; a thromboxane A2 agonist;
an ergot alkaloid; a compound which modulates the action of a
naturetic factor, particularly a compound which modulates the
action of atrial naturetic factor (also known as atrial naturetic
peptide), B type and C type naturetic factors; an angiotensin
receptor antagonist; a substrate for NO-synthase; a calcium channel
blocker; an antagonist of endothelin receptors; an inhibitor of
endothelin converting enzyme; a cholesterol lowering agent such as
an HMG-CoA reductase inhibitor; an antiplatelet or antithrombotic
agent; an insulin sensitizing agent such as a glitazone; an insulin
secretagogue such as a sulfonylurea; an acetylcholinesterase
inhibitor; an estrogen receptor modulator; a PDE5 inhibitor; a
PDE11 inhibitor; a neuropeptide Y (NPY) inhibitor, preferably an
NPY5 inhibitor and even more preferably an NPY1 inhibitor, said NPY
inhibitor having an IC50 of less than 100 nM, and more preferably
an IC50 of less than 50 nM; a vasoactive intestinal protein (VIP)
or a VIP mimetic, more particularly a VIP which is mediated by one
or more of the VIP receptor subtypes VPAC1, VPAC or PACAP
(pituitary adenylate cyclase activating peptide); a VIP receptor
agonist; a VIP analogue or fragment; an .alpha.-adrenoreceptor
antagonist/VIP combination (e.g., Invicorp.RTM., Aviptadil); a
serotonin receptor agonist, antagonist or modulator, more
particularly, a modulator for 5HT1A; a testosterone replacement
agent; estrogen; a combination of estrogen and medroxyprogesterone;
a combination of estrogen and medroxyprogesterone acetate (MPA); a
combination of estrogen and a methyl testosterone hormone
replacement therapy agent (e.g., HRT); a modulator of transporters
for noradrenaline, dopamine or serotonin; a purinergic receptor
agonist or modulator; a neurokinin (NK) receptor antagonist; an
opioid receptor agonist, antagonist or modulator, preferably an
agonist for the ORL-1 receptor; an oxtocin/vasopressin receptor
modulator or agonist, preferably a selective oxytocin agonist or
modulator; a cannabinoid receptor modulator; a central nervous
system (CNS) active agent; an angiotensin-converting enzyme
inhibitor; a combinatino of an angiotensin converting-enzyme
inhibitor and a neutral endopeptidase; L-Dopa; a combination of
L-Dopa and carbidopa; a steroidal anti-inflammatory agent; a
non-steroidal anti-inflammatory agent; a proten kinase C-.beta.
inhibitor; an AMP-activated protein kinase activator; insulin; a
weight loss agent; a dipeptidyl peptidase IV (DPP IV) inhibitor; a
glucagon antagonist; an I kappa B kinase-.beta. (IKK-.beta.)
inhibitor such as salicylate; a PTP1B inhibitor; an agent that
reduces the levels of PTP1B levels using antisense technology; a
glycogen synthase kinase-3 inhibitor; a GLP-1 agonist; a
PPAR-.gamma. agonist; a PPAR-.gamma. antagonist; a PPAR-.alpha.
agonist; a dual PPAR-.alpha./PPAR-.gamma. agonist; a RXR
antagonist; a biguanide such as metformin, a glycogen phosphorylase
inhibitor; a sorbitol dehydrogenase inhibitor (SDI); an aldose
reductase inhibitor (ARI); a soluble guanylate cyclase (sGC)
activator; growth hormone; or a growth hormone secretagogue.
[0101] Any naturally occurring or synthetic prostaglandin or ester
thereof may be used as the second compound of a combination of this
invention. Suitable prostaglandins for use herein include
alprostadil, prostaglandin E.sub.1, prostaglandin E.sub.0,
13,14-dihydroprosta glandin E.sub.1, prostaglandin E.sub.2,
eprostinol, natural synthetic and semi-synthetic prostaglandins and
derivatives thereof including those described in International
Patent Application Publication No. WO00/33825 and U.S. Pat. No.
6,037,346; PGE.sub.0, PGE.sub.1, PGA.sub.1, PGB.sub.1,
PGF.sub..alpha., 19-hydroxy PGA.sub.1, 19-hydroxy--PGB.sub.1,
PGE.sub.2, PGB.sub.2, 19-hydroxy-PGA.sub.2, 19-hydroxy-PGB.sub.2,
PGE.sub.3.alpha., carboprost tromethamine dinoprost, tromethamine,
dinoprostone, lipoprost, gemeprost, metenoprost, sulprostune,
tiaprost and moxisylate.
[0102] The disclosures made in U.S. patents, International patent
applications and all other references mentioned herein are hereby
incorporated by reference.
[0103] Any .alpha.-adrenergic receptor antagonist compound may be
used as the second compound of a combination of this invention.
Suitable .alpha.-adrenergic receptor antagonists for use herein
include the .alpha.-adrenergic receptor blockers described in
International Patent Application Publication No. WO99/30697.
Selective .alpha..sub.1-adrenocep- tor, .alpha..sub.2-adrenoceptor
blockers and non-selective adrenoceptor blockers may also be used
as the second .alpha.-adrenergic receptor antagonist compound of
this invention. Suitable .alpha..sub.1-adrenocepto- r blockers
include phentolamine, phentolamine mesylate, trazodone, alfuzosin,
indoramin, naftopidil, tamsulosin, dapiprazole, phenoxybenzamine,
idazoxan, efaraxan, yohimbine, rauwolfa alkaloids, Recordati
15/2739, SNAP 1069, SNAP 5089, RS17053, SL 89.0591, doxazosin,
terazosin, abanoquil and prazosin. Suitable
.alpha..sub.2-adrenoceptor blockers include those disclosed in U.S.
Pat. No. 6,037,346, dibenamine, tolazoline, trimazosin and
dibenamine. Suitable .alpha.-adrenergic receptors for use as the
second compound of a combination of this invention are also
described in U.S. Pat. Nos. 4,188,390; 4,026,894; 3,511,836;
4,315,007; 3,527,761; 3,997,666; 2,503,059; 4,703,063; 3,381,009;
4,252,721 and 2,599,000. Other suitable .alpha..sub.2-adrenoceptor
blockers include clonidine, papaverine, papaverine hydrochloride,
each of which may optionally be administered in the presence of a
cariotonic agent such as, but not limited to, pirxamine.
[0104] Any nitric oxide donor (NO-donor or NO-agonist) compound may
be used as the second compound of a combination of this invention.
Suitable NO-donor compounds for use herein include organic
nitrates, such as mono-, di- or tri-nitrates; organic nitrate
esters such as glyceryl binitrate (also known as nitroglycerin),
isosorbide 5-mononitrate, isosorbide dinitrate, pentaerythritol
tetranitrate, erythrityl tetranitrate, amylnitrate, a diazenium
diolate (NONOate), and 1,5-pentanedinitrate; sodium nitroprusside
(SNP); 3-morpholinosydnonimine molsidomine; S-nitroso-N-acetyl
penicilliamine (SNAP); S-nitroso-N-glutathione (SNO-GLU);
N-hydroxy-L-arginine; linsidomine; linsidomine chlorohydrate;
(SIN-1) S-nitroso-N-cysteine; L-arginine; ginseng; zizphi fructus;
molsidomine; Re-2047; and nitrosylated maxisylyte derivatives such
as NMI-678-11 and NMI-937 (International Patent Application
Publication No. WO00/12075).
[0105] Any potassium channel opener or modulator may be used as the
second compound of a combination of this invention. Suitable
potassium channel openers/modulators for use herein include
nicorandil, cromokalim, levcromakalim, lemakalim, pinacidil,
cliazoxide, minoxidil, charybdotoxin, glyburide, glipizide,
4-aminipyridine and barium chloride (BaCl.sub.2).
[0106] Any dopaminergic agent may be used as the second compound of
a combination of this invention. Preferred dopaminergic agents
include apomorphine and selective D2, D3 and D2/D.sub.3agonists
such as pramipexole, ropirinol (International Patent Application
Publication No. WO00/23056), L-Dopa, L-Dopa in combination with
carbidopa, PNU95666 (International Patent Application Publication
No. WO00/40226).
[0107] Any vasodilator agent may be used as the second compound of
a combination of this invention. Suitable vasodilator agents for
use herein include nimodepine, pinacidil, cyclandelate,
isoxsuprine, chloroprumazine, haloperidol, Rec 15/2739 and
trazodone.
[0108] Any ergot alkoloid may be used as the second compound of a
combination of this invention. Suitable ergot alkaloids include
those disclosed in U.S. Pat. No. 6,037,346; acetergamine,
brazergoline, bromerguride, cianergoline, delorgotrile,
disulergine, ergonovine maleate, ergotamine tartrate, etisulergine,
lergotrile, lysergide, mesulergine, metergoline, metergotamine,
nicergoline, pergolide, propisergide, proterguride, terguride.
[0109] Any angiotensin receptor antagonist may be used as the
second compound of a combination of this invention. Suitable
angiotensin receptor antagonists include losartan, candersartan,
eprosartan, irbesartan and valsartan.
[0110] Any substrate for NO-synthase may be used as the second
compound of a combination of this invention. Suitable NO-synthase
substrates include, inter alia, L-arginine.
[0111] Any calcium channel blocker may be used as the second
compound of a combination of this invention. Suitable calcium
channel blockers include, amlodipine (amlodipine besylate is also
known as Norvasc.RTM.), bepridil, which may be prepared as
disclosed in U.S. Pat. No. 3,962,238 or U.S. Reissue No. 30,577;
clentiazem, which may be prepared as disclosed in U.S. Pat. No.
4,567,175; diltiazem, which may be prepared as disclosed in U.S.
Pat. No. 3,562, fendiline, which may be prepared as disclosed in
U.S. Pat. No. 3,262,977; gallopamil, which may be prepared as
disclosed in U.S. Pat. No. 3,261,859; mibefradil, which may be
prepared as disclosed in U.S. Pat. No. 4,808,605; prenylamine,
which may be prepared as disclosed in U.S. Pat. No. 3,152,173;
semotiadil, which may be prepared as disclosed in U.S. Pat. No.
4,786,635; terodiline, which may be prepared as disclosed in U.S.
Pat. No. 3,371,014; verapamil, which may be prepared as disclosed
in U.S. Pat. No. 3,261,859; aranipine, which may be prepared as
disclosed in U.S. Pat. No. 4,572,909; barnidipine, which may be
prepared as disclosed in U.S. Pat. No. 4,220,649; benidipine, which
may be prepared as disclosed in European Patent Application
Publication No. 106,275; cilnidipine, which may be prepared as
disclosed in U.S. Pat. No. 4,672,068; efonidipine, which may be
prepared as disclosed in U.S. Pat. No. 4,885,284; elgodipine, which
may be prepared as disclosed in U.S. Pat. No. 4,952,592;
felodipine, which may be prepared as disclosed in U.S. Pat. No.
4,264,611; isradipine, which may be prepared as disclosed in U.S.
Pat. No. 4,466,972; lacidipine, which may be prepared as disclosed
in U.S. Pat. No. 4,801,599; lercanidipine, which may be prepared as
disclosed in U.S. Pat. No. 4,705,797; manidipine, which may be
prepared as disclosed in U.S. Pat. No. 4,892,875; nicardipine,
which may be prepared as disclosed in U.S. Pat. No. 3,985,758;
nifedipine, which may be prepared as disclosed in U.S. Pat. No.
3,485,847; nilvadipine, which may be prepared as disclosed in U.S.
Pat. No. 4,338,322; nimodipine, which may be prepared as disclosed
in U.S. Pat. No. 3,799,934; nisoldipine, which may be prepared as
disclosed in U.S. Pat. No. 4,154,839; nitrendipine, which may be
prepared as disclosed in U.S. Pat. No. 3,799,934; cinnarizine,
which may be prepared as disclosed in U.S. Pat. No. 2,882,271;
flunarizine, which may be prepared as disclosed in U.S. Pat. No.
3,773,939; lidoflazine, which may be prepared as disclosed in U.S.
Pat. No. 3,267,104; lomerizine, which may be prepared as disclosed
in U.S. Pat. No. 4,663,325; bencyclane, which may be prepared as
disclosed in Hungarian Patent No. 151,865; etafenone, which may be
prepared as disclosed in German Patent No. 1,265,758; and
perhexiline, which may be prepared as disclosed in British Patent
No. 1,025,578.
[0112] Any one cholesterol lowering agent may be used as the second
compound of a combination of this invention. Suitable cholesterol
lowering agents include vastatins such as simvastatin, disclosed in
U.S. Pat. No. 4,444,784; pravastatin, disclosed in U.S. Pat. No.
4,346,227; cerivastatin, disclosed in U.S. Pat. No. 5,502,199;
mevastatin, disclosed in U.S. Pat. No. 3,983,140; velostatin,
disclosed in U.S. Pat. No. 4,448,784 and U.S. Pat. No. 4,450,171;
fluvastatin, disclosed in U.S. Pat. No. 4,739,073; compactin,
disclosed in U.S. Pat. No. 4,804,770; lovastatin, disclosed in U.S.
Pat. No. 4,231,938; dalvastatin, disclosed in European Patent
Application Publication No. 738510 A2; fluindostatin, disclosed in
European Patent Application Publication No. 363934 A1;
atorvastatin, disclosed in U.S. Pat. No. 4,681,893; atorvastatin
calcium (atorvastatin calcium is also known as Lipitor.RTM.),
disclosed in U.S. Pat. No. 5,273,995; and dihydrocompactin,
disclosed in U.S. Pat. No. 4,450,171. Other suitable cholesterol
lowering agents include fibrates.
[0113] Any antiplatelet and antithrombotic agent may be used as the
second compound of a combination of this invention. Suitable
antiplatelet and antithrombotic agents include, e.g., tPA, uPA,
warfarin, hirudin and other thrombin inhibitors, heparin and
thromboplastin activating factor inhibitors.
[0114] Any insulin sensitising agent may be used as the second
compound of a combination of this invention. Suitable insulin
sensitizing agents include Avandia.RTM., Actos.RTM. and
hypoglycaemic agents such as, but not limited to, sulfonylureas
such as glipizide, metformin and acarbose.
[0115] Any acetylcholinesterase inhibitor may be used as the second
compound of a combination of this invention. A suitable
acetylcholinesterase inhibitor is, e.g., donezipil.
[0116] Any-estrogen receptor modulator, estrogen agonist or
estrogen antagonist may be used as the second compound of a
combination of this invention. Suitable estrogen receptor
modulators, estrogen agonists or estrogen antagonists include the
compounds disclosed in International Patent Application Publication
No. WO96/21656 and U.S. Pat. No. 5,552,412. Preferred such
compounds include raloxifene, lasofoxifene,
(-)-cis-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,8-tetrahyd-
ronaphthalene-2-ol and pharmaceutically acceptable salts
thereof.
[0117] Any PDE5 or PDE11 inhibitor may be used as the second
compound of a combination of this invention. It is particularly
preferred that a PDE5 inhibitor be used as the second compound of
this invention. Suitable PDE5 inhibitors include the
pyrazolo[4,3-d]pyrimidin-7-ones disclosed in EP-A-0463756; the
pyrazolo[4,3-d]pyrimidin-7-ones disclosed in EP-A-0526004; the
pyrazolo[4,3-d]pyrimidin-7-ones disclosed in published
international patent application WO 93/06104; the isomeric
pyrazolo[3,4-d]pyrimidin-4-ones disclosed in International Patent
Application Publication No. WO93/07149; the quinazolin-4-ones
disclosed in International Patent Application Publication No.
WO93/12095; the pyrido[3,2-d]pyrimidin-4-ones disclosed in
International Patent Application Publication No. WO94/05661; the
purin-6-ones disclosed in International Patent Application
Publication No. WO94/00453; the pyrazolo[4,3-d]pyrimidin-7-ones
disclosed in International Patent Application Publication No.
WO98/49166; the pyrazolo[4,3-d]pyrimidin-7-on- es disclosed in
International Patent Application Publication No. WO99/54333; the
pyrazolo[4,3-d]pyrimidin-4-ones disclosed in EP-A-0995751; the
pyrazolo[4,3-d]pyrimidin-7-ones disclosed in International Patent
Application Publication No. WO00/24745; the
pyrazolo[4,3-d]pyrimidin-4-ones disclosed in EP-A-0995750; the
compounds disclosed in International Patent Application Publication
No. WO95/19978; the compounds disclosed in International Patent
Application Publication No. WO99/24433; he
pyrazolo[4,3-d]pyrimidin-7-ones disclosed in International Patent
Application Publication No. WO01/27112; the
pyrazolo[4,3-d]pyrimidin-7-ones disclosed in International Patent
Application Publication No. WO01/27113; the compounds disclosed in
EP-A-1092718; the compounds disclosed in EP-A-1092719; and the
compounds disclosed in International Patent Application Publication
No. WO93/07124.
[0118] Preferred PDE5 inhibitors for use as a second compound in a
combination of this invention include:
5-[2-ethoxy-5-(4-methyl-1-piperazi-
nylsulphonyl)phenyl]-1-methyl-3-n-propyl-1,6-dihydro-7H-pyrazolo[4,3-d]pyr-
imidin-7-one (sildenafil) also known as
1-[[3-(6,7-dihydro-1-methyl-7-oxo--
3-propyl-1H-pyrazolo[4,3-d]pyrimidin-5-yl)-4-ethQxyphenyl]sulphonyl]-4-met-
hylpiperazine (see EP-A-0463756);
5-(2-ethoxy-5-morpholinoacetylphenyl)-1--
methyl-3-n-propyl-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one
(see EP-A-0526004);
3-ethyl-5-[5-(4-ethylpiperazin-1-ylsulphonyl)-2-n-propoxyp-
henyl]-2-(pyridin-2-yl)methyl-2,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-on-
e (see WO98/49166);
3-ethyl-5-[5-(4-ethylpiperazin-1-ylsulphonyl)-2-(2-met-
hoxyethoxy)pyridin-3-yl]-2-(pyridin-2-yl)methyl-2,6-dihydro-7H-pyrazolo[4,-
3-d]pyrimidin-7-one (see WO99/54333);
6-benzo[1,3]dioxol-5-yl-2-methyl-2,3-
,6,7,12,12a-hexahydro-pyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione
(cialis);
(+)-3-ethyl-5-[5-(4-ethylpiperazin-1-ylsulphonyl)-2-(2-methoxy-- 1
(R)-methylethoxy)pyridin-3-yl]-2-methyl-2,6-dihydro-7H-pyrazolo[4,3-d]py-
rimidin-7-one, also known as
3-ethyl-5-{5-[4-ethylpiperazin-1-ylsulphonyl]-
-2-([(1R)-2-methoxy-1-methylethyl]oxy)pyridin-3-yl}-2-methyl-2,6-dihydro-7-
H-pyrazolo[4,3-d]pyrimidin-7-one (see WO99/54333);
5-[2-ethoxy-5-(4-ethylp-
iperazin-1-ylsulphonyl)pyridin-3-yl]-3-ethyl-2-[2-methoxyethyl]-2,6-dihydr-
o-7H-pyrazolo[4,3-d]pyrimidin-7-one, also known as
1-{6-ethoxy-5-[3-ethyl--
6,7-dihydro-2-(2-methoxyethyl)-7-oxo-2H-pyrazolo[4,3-d]pyrimidin-5-yl]-3-p-
yridylsulphonyl}-4-ethylpiperazine (see WO01/27113, Example 8);
5-[2-iso-butoxy-5-(4-ethylpiperazin-1-ylsulphonyl)pyridin-3-yl]-3-ethyl-2-
-(1-methylpiperidin-4-yl)-2,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one
(see WO1/27113, Example 15);
5-[2-ethoxy-5-(4-ethylpiperazin-1-ylsulphony-
l)pyridin-3-yl]-3-ethyl-2-phenyl-2,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-
-one (see WO01/27113, Example 66);
5-(5-acetyl-2-propoxy-3-pyridinyl)-3-et-
hyl-2-(1-isopropyl-3-azetidinyl)-2,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-
-one (see WO01/27112, Example 124);
5-(5-acetyl-2-butoxy-3-pyridinyl)-3-et-
hyl-2-(1-ethyl-3-azetidirlyl)-2,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-on-
e (see WO01/27112, Example 132);
(6R,12aR)-2,3,6,7,12,12a-hexahydro-2-meth-
yl-6-(3,4-methylenedioxyphenyl)-pyrazino[2',1':6,1]pyrido[3,4-b]indole-1,4-
-dione (IC-351), i.e. the compound of examples 78 and 95 of
published international application WO95/19978, as well as the
compound of examples 1, 3, 7 and 8;
2-[2-ethoxy-5-(4-ethyl-piperazin-1-yl-1-sulphonyl)-phenyl]-
-5-methyl-7-propyl-3H-imidazo[5,1-f][1,2,4]triazin-4-one
(vardenafil) also known as
1-[[3-(3,4-dihydro-5-methyl-4-oxo-7-propylimidazo[5,1-f]-as-tria-
zin-2-yl)-4-ethoxyphenyl]sulphonyl]-4-ethylpiperazine, i.e. the
compound of examples 20, 19, 337 and 336 of published international
application WO99/24433; the compound of example 11 in WO93/07124
(EISAI); and compounds 3 and 14 from Rotella D P, J. Med. Chem.,
2000, 43, 1257.
[0119] Still other type cGMP PDE5 inhibitors useful in conjunction
with the present invention include:
4-bromo-5-(pyridylmethylamino)-6-[3-(4-chl-
orophenyl)-propoxy]-3(2H)pyridazinone;
1-[4-[(1,3-benzodioxol-5-ylmethyl)a- miono]-6-chloro-2-q
uinozolinyl]-4-piperidine-carboxylic acid, monosodium salt;
(+)-cis-5,6a,7,9,9,9a-hexahydro-2-[4-(trifluoromethyl)-phenylmethyl-
-5-methyl-cyclopent-4,5]imidazo[2,1-b]purin-4(3H)one;
furazlocillin;
cis-2-hexyl-5-methyl-3,4,5,6a,7,8,9,9a-octahydrocyclopent[4,5]-imidazo[2,-
1-b]purin-4-one;
3-acetyl-1-(2-chlorobenzyl)-2-propylindole-6-carboxylate;
3-acetyl-1-(2-chlorobenzyl)-2-propylindole-6-carboxylate;
4-bromo-5-(3-pyridylmethylamino)-6-(3-(4-chlorophenyl)
propoxy)-3-(2H)pyridazinone;
1-methyl-5(5-morpholinoacetyl-2-n-propoxyphe-
nyl)-3-n-propyl-1,6-dihydro-7H-pyrazolo(4,3-d)pyrimidin-7-one;
1-[4-[(1,3-benzodioxol-5-ylmethyl)amino]-6-chloro-2-quinazolinyl]-4-piper-
idinecarboxylic acid, monosodium salt; Pharmaprojects No. 4516
(Glaxo Wellcome); Pharmaprojects No. 5051 (Bayer); Pharmaprojects
No. 5064 (Kyowa Hakko; see WO 96/26940); Pharmaprojects No. 5069
(Schering Plough); GF-196960 (Glaxo Wellcome); E-8010 and E-4010
(Eisai); Bay-38-3045 & 38-9456 (Bayer) and Sch-51866; selected
from: sildenafil,
5-(2-ethoxy-5-morpholinoacetylphenyl)-1-ethyl-3-n-propyl-1,6-dihydro-7H-p-
yrazolo[4,3-d]pyrimidin-7-one,
3-ethyl-5-[5-(4-ethylpiperazin-1-ylsulphony-
l)-2-n-propoxyphenyl]-2-(pyridin-2-yl)methyl-2,6-dihydro-7H-pyrazolo[4,3-d-
]pyrimidin-7-one,
3-ethyl-5-[5-(4-ethylpiperazin-1-ylsulphonyl)-2-(2-metho-
xyethoxy)pyridin-3-yl]-2-(pyridin-2-yl)methyl-2,6-dihydro-7H-pyrazolo[4,3--
d]pyrimidin-7-one,
5-[2-ethoxy-5-(4-ethylpiperazin-1-ylsulphonyl)pyridin-3-
-yl]-3-ethyl-2-(2-methoxyethyl)-2,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7--
one and
1-[[3-(3,4-dihydro-5-methyl-4-oxo-7-propylimidazolo[5,1-f]-as-triz-
in-2-yl)-4-ethoxyphenyl]sulphonyl]-4-ethylpiperzine or a
pharmaceutically acceptable salt, solvate, pro-drug or polymorph
thereof.
[0120] More preferred cGMP PDE5 inhibitors for use as the second
compound in a combination of this invention include sildenafil,
sildenafil citrate (also known as Viagra.RTM.;
5-(5-acetyl-2-butoxy-3-pyridinyl)-3-ethyl-2-(-
1-ethyl-3-azetidinyl)-2,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one;
2-[2-ethoxy-5-(4-ethyl-piperazin-1-yl-1-sulphonyl)-phenyl]-5-methyl-7-pro-
pyl-3H-imidazo[5,1-f][1,2,4]triazin-4-one (vardenafil);
6-benzo[1,3]dioxol-5-yl-2-methyl-2,3,6,7,12,12a-hexahydro-pyrazino[1',2':-
1,6]pyrido[3,4-b]indole-1,4-dione (cialis); and
5-[2-ethoxy-5-(4-ethylpipe-
razin-1-ylsulphonyl)pyridin-3-yl]-3-ethyl-1-(2-methoxyethyl)-1,6-dihydro-7-
H-pyrazolo[4,3-d]pyrimidin-7-one.
[0121] Any melanocortin receptor agonist, melanocortin receptor
modulator or melanocortin receptor enhancer may be used as the
second compound of a combination of this invention. Suitable
melanocortin receptor agonists, modulators or enhancers include
melanotan II; PT-14; PT-141; and compounds disclosed in
International Patent Application Publication Nos. WO99/64002,
WO00/74679, WO99/55679, WO01/05401, WO00/58361, WO01/14879,
WO01/13112 and WO99/54358.
[0122] Any serotonin receptor agonist, antagonist or modulator may
be used as the second compound in a combination of this invention.
It is particularly preferred to use agonists, antagonists or
modulators of 5HT1A. Suitable such agonists, antagonists or
modulators include VML 670; 5HT2A; 5HT2C; 5HT3; and 5HT6 receptors,
including those described in International Patent Application
Publication Nos. WO99/02159, WO0/02550 and WO00/28993.
[0123] Any testosterone replacement agent may be used as the second
compound in a combination of this invention. Suitable testosterone
replacement agents include dehydroandrostendione, testosternone
(Tostrelle), dihydrotestosterone and testosterone implants.
[0124] Any hormone replacement therapy (HRT) agent may be used as
the second compound of a combination of this invention. Suitable
HRT agents include Premarin.RTM., Cenestin.RTM.,
Oestrofeminal.RTM., Equin.RTM., Estrace.RTM., Estrofem.RTM.,
Elleste Solo.RTM., Estring.RTM., Eastraderm TTS.RTM., Eastraderm
Matrix.RTM., Dermestril.RTM., Premphase.RTM., Preempro.RTM.,
Prempak.RTM., Premique.RTM., Estratest.RTM., Estratest HS.RTM. and
Livial.RTM. (tibolone).
[0125] Any modulator of transporters for noradrenaline, dopamine
and/or serotonin may be used as the second compound of a
combination of this invention. Suitable such modulators include
bupropion and GW-320659.
[0126] Any neurokinin (NK) receptor antagonist may be used as the
second compound of a combination of this invention. Suitable NK
receptor antagonists include those described in International
Patent Application Publication No. WO99/64008.
[0127] Any angiotensin converting enzyme inhibitor (ACE inhibitor)
may be used as the second compound of a combination of this
invention. Suitable ACE inhibitors include, but are not limited to:
alacepril, which may be prepared as disclosed in U.S. Pat. No.
4,248,883; benazepril, which may be prepared as disclosed in U.S.
Pat. No. 4,410,520; captopril, which may be prepared as disclosed
in U.S. Pat. Nos. 4,046,889 and 4,105,776; ceronapril, which may be
prepared as disclosed in U.S. Pat. No. 4,452,790; delapril, which
may be prepared as disclosed in U.S. Pat. No. 4,385,051; enalapril,
which may be prepared as disclosed in U.S. Pat. No. 4,374,829;
fosinopril, which may be prepared as disclosed in U.S. Pat. No.
4,337,201; imadapril, which may be prepared as disclosed in U.S.
Pat. No. 4,508,727; lisinopril, which may be prepared as disclosed
in U.S. Pat. No. 4,555,502; moveltopril, which may be prepared as
disclosed in Belgian Patent No. 893,553; perindopril, which may be
prepared as disclosed in U.S. Pat. No. 4,508,729; quinapril, which
may be prepared as disclosed in U.S. Pat. No. 4,344,949; ramipril,
which may be prepared as disclosed in U.S. Pat. No. 4,587,258;
spirapril, which may be prepared as disclosed in U.S. Pat. No.
4,470,972; temocapril, which may be prepared as disclosed in U.S.
Pat. No. 4,699,905; and trandolapril, which may be prepared as
disclosed in U.S. Pat. No. 4,933,361.
[0128] Any compound which is a combined inhibitor of
angiotensin-converting enzyme and neutral endopeptidase may be used
as the second compound of a combination of this invention. A
suitable such combined inhibitor is, e.g., omapatrilat.
[0129] Any protein kinase C-.beta. inhibitor may be used as the
second compound of a combination of this invention. A suitable
protein kinase C-.beta. inhibitor is, e.g., LY333531.
[0130] Any activator of AMP-activated protein kinase may be used as
the second compound of a combination of this invention. A suitable
such activator is, e.g., 5-amino-4-imidazolecarboxamide
ribonucleoside.
[0131] Any weight loss agent may be used as the second compound of
a combination of this invention. Suitable weight loss agents
include sibutramine and orlistat.
[0132] Any dipeptidyl peptidase IV (DPPIV) inhibitor may be used as
the second compound of a combination of this invention. Suitable
DPPIV inhibitors include NVP DPP728 and P32/98.
[0133] Any glucagon antagonist may be used as the second compound
of a combination of this invention. A suitable glucagon antagonist
is, e.g., NNC25-2504.
[0134] Any IKK-.beta. inhibitor may be used as the second compound
of a combination of this invention. A suitable IKK-.beta. inhibitor
is, e.g., salicylate.
[0135] Any PTP1B inhibitor may be used as the second compound of a
combination of this invention. A suitable PTP1 B inhibitor is,
e.g., PTP112.
[0136] Any glycogen synthase kinase-3 (GSK-3) inhibitor may be used
as the second compound of a combination of this invention. A
suitable GSK-3 inhibitor is, e.g., Chir98014.
[0137] Any GLP-1 agonist may be used as the second compound of a
combination of this invention. Suitable GLP-1 agonists include
GLP1, NN-2211 and exendin 4.
[0138] Any PPAR-.gamma. agonist may be used as the second compound
of a combination of this invention. Suitable PPAR-.gamma. agonists
include Rezulin.RTM., Avandia.RTM., Actos.RTM. or CS011.
[0139] Any PPAR-.gamma. antagonist may be used as the second
compound of a combination of this invention. A suitable
PPAR-.gamma. antagonist is, e.g., bisphenol A diglycidyl ether
(BADGE).
[0140] Any PPAR-.alpha. agonist may be used as the second compound
of a combination of this invention. A suitable PPAR-.alpha. agonist
is, e.g., fenofibrate.
[0141] Any dual PPAR-.alpha./PPAR-.gamma. agonist may be used as
the second compound of a combination of this invention. Suitable
such dual agonists include farglitazar, GW1929, DRF2725, AZ242 and
KRP 297.
[0142] Any RXR antagonist may be used as the second compound of a
combination of this invention. A suitable RXR antagonist is, e.g.,
HX531.
[0143] Any glycogen phosphorylase inhibitor may be used as the
second compound of a combination of this invention. A suitable
glycogen phosphorylase inhibitor is, e.g., CP-316819.
[0144] Any sorbitol dehydrogenase inhibitor (SDI) may be used as
the second compound of a combination of this invention. Suitable
SDIs include those dislcosed in International Patent Application
Publication No. WO00/59510. A particuarly preferred SDI is
1R-(4-(4-(4,6-dimethyl)-[1,3,5-
]triazin-2-yl)-2R,6S-dimethyl-piperazin-1-yl)-pyrimidin-2-yl)-ethanol.
[0145] Any aldose reductase inhibitor (ARI) may be used as the
second compound of a combination of this invention. Suitable ARIs
include zopolrestat, epalrestat, ponalrestat, zenarestat or
fidarestat.
[0146] Other suitable ARIs for use as the second compound in a
combination of this invention include compounds of the Formula ARI
3
[0147] prodrugs thereof and pharmaceutically acceptable salts of
said compounds and said prodrugs, wherein:
[0148] A is S, SO or SO.sub.2;
[0149] R.sup.1 and R are each independently hydrogen or methyl;
[0150] R.sup.3 is Het.sup.1, --CHR.sup.4Het.sup.1 or
NR.sup.6R.sup.7;
[0151] R.sup.4 is hydrogen or (C.sub.1-C.sub.3)alkyl;
[0152] R.sup.6 is (C.sub.1-C.sub.6)alkyl, aryl or Het.sup.2;
[0153] R.sup.7 is Het.sup.3;
[0154] Het.sup.1 is pyridyl, pyrimidyl, pyrazinyl, pyridazinyl,
quinolyl, isoquinolyl, quinazolyl, quinoxalyl, phthalazinyl,
cinnolinyl, naphthyridinyl, pteridinyl, pyrazinopyrazinyl,
pyrazinopyridazinyl, pyrimidopyridazinyl, pyrimidopyrimidyl,
pyridopyrimidyl, pyridopyrazinyl, pyridopyridazinyl, pyrrolyl,
furanyl, thienyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl,
isoxazolyl, isothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl,
tetrazolyl, indolyl, benzofuranyl, benzothienyl, benzimidazolyl,
benzoxazolyl, benzothiazolyl, indazolyl, benzisoxazolyl,
benzisothiazolyl, pyrrolopyridyl, furopyridyl, thienopyridyl,
imidazolopyridyl, oxazolopyridyl, thiazolopyridyl, pyrazolopyridyl,
isoxazolopyridyl, isothiazolopyridyl, pyrrolopyrimidyl,
furopyrimidyl, thienopyrimidyl, imidazolopyrimidyl,
oxazolopyrimidyl, thiazolopyrimidyl, pyrazolopyrimidyl,
isoxazolopyrimidyl, isothiazolopyrimidyl, pyrrolopyrazinyl,
furopyrazinyl, thienopyrazinyl, imidazolopyrazinyl,
oxazolopyrazinyl, thiazolopyrazinyl, pyrazolopyrazinyl,
isoxazolopyrazinyl, isothiazolopyrazinyl, pyrrolopyridazinyl,
furopyridazinyl, thienopyridazinyl, imidazolopyridazinyl,
oxazolopyridazinyl, thiazolopyridazinyl, pyrazolopyridazinyl,
isoxazolopyridazinyl or isothiazolopyridazinyl; Het.sup.1 is
optionally substituted with up to a total of four substituents each
independently selected from halo, formyl,
(C.sub.1-C.sub.6)alkoxycarbonyl,
(C.sub.1-C.sub.6)alkylenyloxycarbonyl,
(C.sub.1-C.sub.4)alkoxy-(C.sub.1-C- .sub.4)alkyl,
C(OH)R.sup.12R.sup.13, (C.sub.1-C.sub.4)alkylcarbonylamido,
(C.sub.3-C.sub.7)cycloalkylcarbonylamido, phenylcarbonylamido,
benzyl, phenyl, naphthyl, imidazolyl, pyridyl, triazolyl,
benzimidazolyl, oxazolyl, isoxazolyl, thiazolyl, oxadiazolyl,
thiadiazolyl, tetrazolyl, thienyl, benzothiazolyl, pyrrolyl,
pyrazolyl, quinolyl, isoquinolyl, benzoxazolyl, pyridazinyl,
pyridyloxy, pyridylsulfonyl, furanyl, phenoxy, thiophenoxy,
(C.sub.1-C.sub.4)alkylsulfenyl, (C.sub.1-C.sub.4)alkylsulfon- yl,
(C.sub.3-C.sub.7)cycloalkyl, (C.sub.1-C.sub.6)alkyl optionally
substituted with up to three fluoro, or (C.sub.1-C.sub.4)alkoxy
optionally substituted with up to five fluoro; said benzyl, phenyl,
naphthyl, imidazolyl, pyridyl, triazolyl, benzimidazolyl, oxazolyl,
isoxazolyl, thiazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl,
thienyl, benzothiazolyl, pyrrolyl, pyrazolyl, quinolyl,
isoquinolyl, benzoxazolyl, pyridazinyl, pyridyloxy,
pyridylsulfonyl, furanyl, phenoxy, thiophenoxy, in the definition
of substituents for Het.sup.1 are optionally substituted with up to
three substituents independently selected from hydroxy, halo,
hydroxy-(C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkoxy-(C-
.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.6)alkylsulfenyl,
(C.sub.1-C.sub.6)alkylsulfinyl, (C.sub.1-C.sub.6)alkylsulfonyl,
(C.sub.1-C.sub.6)alkyl optionally substituted with up to five
fluoro and (C.sub.1-C.sub.6)alkoxy optionally substituted with up
to five fluoro; said imidazolyl, oxazolyl, isoxazolyl, thiazolyl
and pyrazolyl in the definition of substituents for Het.sup.1 are
optionally substituted with up to two substituents independently
selected from hydroxy, halo, C.sub.1-C.sub.6)alkyl,
hydroxy-(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy-(C.sub.1-C.sub.4)alkyl,
C.sub.1-C.sub.4)alkyl-phe- nyl optionally substituted in the phenyl
portion with one Cl, Br, OMe, Me or SO.sub.2-phenyl wherein said
SO.sub.2-phenyl is optionally substituted in the phenyl portion
with one Cl, Br, OMe, Me, (C.sub.1-C.sub.4)alkyl optionally
substituted with up to five fluoro, or (C.sub.1-C.sub.4)alkoxy
optionally substituted with up to three fluoro;
[0155] R.sup.12 and R.sup.13 are each independently hydrogen or
(C.sub.1-C.sub.4)alkyl;
[0156] Het.sup.2 and Het.sup.3 are each independently imidazolyl,
pyridyl, triazolyl, benzimidazolyl, oxazolyl, isoxazolyl,
thiazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, thienyl,
benzothiazolyl, pyrrolyl, pyrazolyl, quinolyl, isoquinolyl,
benzoxazolyl, pyridazinyl, pyridyloxy, pyridylsulfonyl, furanyl,
phenoxy, thiophenoxy; Het.sup.2 and Het.sup.3 are each
independently optionally substituted with up to a total of four
substituents each independently selected from halo, formyl,
(C.sub.1-C.sub.6)alkoxycarbonyl,
(C.sub.1-C.sub.6)alkylenyloxycarbonyl,
(C.sub.1-C.sub.4)alkoxy-(C.sub.1-C.sub.4)alkyl,
C(OH)R.sup.18R.sup.19, (C.sub.1-C.sub.4)alkylcarbonylamido,
(C.sub.3-C.sub.7)cycloalkylcarbonyla- mido, phenylcarbonylamido,
phenyl, naphthyl, imidazolyl, pyridyl, triazolyl, benzimidazolyl,
oxazolyl, isoxazolyl, thiazolyl, oxadiazolyl, thiadiazolyl,
tetrazolyl, thienyl, benzothiazolyl, pyrrolyl, pyrazolyl, quinolyl,
isoquinolyl, benzoxazolyl, pyridazinyl, pyridyloxy,
pyridylsulfonyl, furanyl, phenoxy, thiophenoxy,
(C.sub.1-C.sub.4)alkylsul- fenyl, (C.sub.1-C.sub.4)alkylsulfonyl,
(C.sub.3-C.sub.7)cycloalkyl, (C.sub.1-C.sub.4)alkyl optionally
substituted with up to three fluoro or (C.sub.1-C.sub.4)alkoxy
optionally substituted with up to five fluoro; said phenyl,
naphthyl, imidazolyl, pyridyl, triazolyl, benzimidazolyl, oxazolyl,
isoxazolyl, thiazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl,
thienyl, benzothiazolyl, pyrrolyl, pyrazolyl, quinolyl,
isoquinolyl, benzoxazolyl, pyridazinyl, pyridyloxy,
pyridylsulfonyl, furanyl, phenoxy, thiophenoxy, in the definition
of substituents for Het.sup.2 and Het.sup.3 are optionally
substituted with up to three substituents independently selected
from hydroxy, halo, hydroxy-(C.sub.1-C.sub.4)alkyl- ,
(C.sub.1-C.sub.4)alkoxy-(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkyl optionally substituted with up to five
fluoro and (C.sub.1-C.sub.4)alkoxy optionally substituted with up
to five fluoro; said imidazolyl, oxazolyl, isoxazolyl, thiazolyl
and pyrazolyl in the definition of substituents for Het.sup.2 and
Het.sup.3 are optionally substituted with up to two substituents
independently selected from hydroxy, halo,
hydroxy-(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy-(C.sub.1-C.sub.4)- alkyl,
(C.sub.1-C.sub.4)alkyl optionally substituted with up to five
fluoro and (C.sub.1-C.sub.4)alkoxy optionally substituted with up
to three fluoro; and
[0157] R.sup.18 and R.sup.19 are each independently hydrogen or
(C.sub.1-C.sub.4)alkyl, provided that when R.sup.3 is
NR.sup.6R.sup.7, then A is SO.sub.2. A particularly preferred
compound of the Formula ARI for use as the second compound of a
combination of this invention is
6-(5-chloro-3-methyl-benzofuran-2-sulfonyl)-2H-pyridazin-3-one.
[0158] Any soluble guanylate cyclase (sGC) activator may be used as
the second compound of a combination of this invention. Suitable
sGC activators include BAY 41-2272 and BAY 41-8543.
[0159] Any growth hormone secretagogue may be used as the second
compound of a combination of this invention. Suitable growth
hormone secretagogues include those disclosed in U.S. Pat. Nos.
6,124,264; 6,110,932; 6,278,000; and 6,251,902. A particularly
preferred growth hormone secretagogue is
2-amino-N-(2-(3a(R)-benzyl-2-methyl-3-oxo-2,3,3a,4,6,7-he-
xahydro-pyrazolo-[4,3c]pyridin-5-yl)-1(R)-benzyloxymethyl-2-oxo-ethyl)-iso-
butyramide.
[0160] Particularly preferred compounds for use as the second
compound in the combinations and pharmaceutical compositions for
use according to the present invention include compounds selected
from the following classes of compounds: insulin sensitizing
agents, PDE5 inhibitors, protein kinase C-.beta. inhibitors,
AMP-activated protein kinase activators, insulin, weight loss
agents, PPAR-.gamma. agonists, PPAR-.alpha. agonists, dual
PPAR-.gamma./PPAR-.alpha. agonists, sorbitol dehydrogenase
inhibitors and aldose reductase inhibitors, each as described
above.
PDE9 Inhibitor--Test Methods
[0161] Phosphodiesterase (PDE) Inhibitory Activity
[0162] Preferred PDE compounds suitable for use in accordance with
the present invention are potent and cGMP PDE9 inhibitors. In vitro
PDE inhibitory activities against cyclic guanosine
3',5'-monophosphate (cGMP) and cyclic adenosine 3',5'-monophosphate
(cAMP) phosphodiesterases are determined by measurement of their
IC.sub.50 values (the concentration of compound required for 50%
inhibition of enzyme activity).
[0163] Phosphodiesterase 9 can be generated from full length human
recombinant clones transfected into SF9 cells as described in
Fisher et al., Journal of Biological Chemistry, 1998, 273,
15559-15564.
[0164] Assays are performed either using a modification of the
"batch" method of W. J. Thompson et al. (Biochem., 1979, 18, 5228)
or using a scintillation proximity assay for the direct detection
of AMP/GMP using a modification of the protocol described by
Amersham plc under product code TRKQ7090/7100. In summary, the
effect of PDE9 inhibitors is investigated by assaying a fixed
amount of enzyme in the presence of varying inhibitor
concentrations and low substrate, (cGMP in a 3:1 ratio unlabelled
to [.sup.3H]-labeled at a concentration of about 1/3 K.sub.m) such
that IC.sub.50.congruent.K.sub.i. The final assay volume is made up
to 100 .mu.l with assay buffer [20 mM Tris-HCl pH 7.4, 5 mM
MgCl.sub.2, 1 mg/ml bovine serum albumin]. Reactions are initiated
with enzyme, incubated for 30-60 minutes at 30.degree. C. to give
<30% substrate turnover and terminated with 50 .mu.l yttrium
silicate SPA beads (containing 3 mM of the respective unlabelled
cyclic nucleotide for PDEs 9 and 11). Plates are re-sealed and
shaken for 20 minutes, after which the beads are allowed to settle
for 30 minutes in the dark and then counted on a TopCount plate
reader (Packard, Meriden, Conn.) Radioactivity units are converted
to percent activity of an uninhibited control (100%), plotted
against inhibitor concentration and inhibitor IC.sub.50 values
obtained using the `Fit Curve` Microsoft Excel extension.
[0165] Effect of Specific PDE9 Inhibitors on Insulin Resistance
Syndrome in Animals--Effects on Plasma Glucose, Triglyceride,
Insulin, and cGMP Levels in ob/ob Mice.
[0166] Biological Data
[0167] Experimental Protocol
[0168] Test Compounds:
[0169] The PDE9 inhibitor compounds to be tested are solubilized in
10% DMSO/0.1% pluronics and dosed via oral gavage using mouse oral
feeding needles (20 gauge, Popper & Sons, Inc., New Hyde Park,
N.Y.). A volume of 4 ml/kg weight is administered for each dose.
Compounds are tested at doses ranging from 1-50 mg/kg.
[0170] Experimental Animals:
[0171] Male ob/ob mice obtained from Jackson Laboratories (Bar
Harbor, Me.) are used in the studies at 6 to 10 weeks of age. Mice
are housed five per cage and allowed free access to D11 mouse chow
(Purina, Brentwood, Mo.) and water.
[0172] Experimental Protocol:
[0173] Mice are allowed to acclimate to the Pfizer animal
facilities for one week prior to the start of the study. On day
one, retro-orbital blood samples are obtained and plasma glucose is
determined as described hereinafter. Mice are then sorted into
groups of five such that mean plasma glucose concentrations for
each group do not differ. On day one, mice are dosed with vehicle
or a test PDE9 inhibitor compound only in the afternoon.
Subsequently, mice are dosed twice a day on day 2-4 in the morning
and in the afternoon. On day five, the mice receive an a.m. dose
and are bled 3 hours later for plasma preparation for glucose and
triglyceride analysis as described below. Terminal plasma samples
are collected on day five following the retro-orbital sinus bleed
as described below. Body weight is measured on days one and five of
the study, and food consumption is assessed over the five day
period.
[0174] Terminal Bleed and Tissue Collection:
[0175] On the morning of the last day of the study mice are dosed
with test compound or vehicle at approximately 8:00 am. Three hours
after dosing, 25 .mu.L of blood is obtained via the retro-orbital
sinus and is added to 100 .mu.L of 0.025 percent heparinized-saline
in Denville Scientific microtubes. The tubes are spun at the
highest setting in a Beckman Microfuge 12 for 2 minutes. Plasma is
collected for plasma glucose and triglyceride determination. The
mice are then sacrificed by decapitation and about one milliliter
of blood is collected in Becton-Dickinson Microtainer brand plasma
separator tubes with lithium heparin. The tubes are spun in a
Beckman Microfuge 12 at the maximum setting for five minutes.
Plasma is collected in 1.5 ml Eppendorf tubes and snap frozen in
liquid nitrogen. Plasma samples are stored at -800.degree. C. until
analyzed.
[0176] Metabolite and Hormone Analysis:
[0177] Plasma glucose and triglycerides are measured using the
Alcyon Clinical Chemistry Analyzer (Abbott Laboratories, Abbott
Park, Ill.) using kits supplied by Abbott. Plasma cGMP is measured
using the Biotrak enzyme-immunoassay system by Amersham
(Piscataway, N.J.). Via a similar technique the plasma insulin is
assessed by the Mercodia ELISA Insulin kit by ALPCO (Uppsala,
Sweden). All assays are conducted according to instructions
provided by the manufacturers.
[0178] Results
[0179] Table 1 illustrates the changes in plasma glucose,
triglyceride, and insulin levels over a five day period observed
with Compound A,
5-(3-chloro-benzyl)-3-isopropyl-1,6-dihydro-pyrazolo[4,3-d]pyrimidin-7-on-
e.
[0180] Taken together, these experimental results in the
hyperglycemic, insulin-resistant ob/ob mouse suggest that selective
PDE9 inhibition improves metabolic parameters associated with
IRS.
1TABLE 1 Plasma Glucose Plasma Triglyceride Plasma Insulin
Treatment (mg/dl) (mg/dl) (pmol/ml) Vehicle 370 .+-. 23 207 .+-. 9
12.0 .+-. 1.5 Compound A 304 .+-. 17 155 .+-. 8 8.2 .+-. 1.5 (10
mg/kg)
[0181] The data in Table 1 are presented as mean.+-.standard error
of the mean.
[0182] Table 2 illustrates the elevation of plasma cGMP produced by
five day treatment with
5-(3-chloro-benzyl)-3-isopropyl-1,6-dihydro-pyrazolo[4-
,3-d]pyrimidin-7-one.
2 TABLE 2 Plasma cGMP Treatment (pmol/ml) Vehicle 9.8 .+-. 0.5
Compound A 16.8 .+-. 3.1 (10 mg/kg)
[0183] The data in Table 2 are presented as mean.+-.standard error
of the mean.
[0184] General Procedure for the Preparation of Examples 1 to 77
4
[0185] The carboxylic acids (80 .mu.mol) were dissolved in a 3.75%
solution of triethylamine in dimethylacetamide (400 .mu.l) and
administered to a 96 well plate. Carbonyldiimidazole (13 mg, 80
.mu.mol) dissolved in pyridine (212 .mu.l) was then added into each
well, and the plates were left to stand at room temperature for 2
hours. A solution of 4-amino-5-isopropyl-2H-pyrazole-3-carboxylic
acid amide (13.5 mg, 80 .mu.mol) dissolved in dimethylacetamide
(100 .mu.l) was then added, and the plates were sealed and heated
to 70.degree. C. in an oven under nitrogen. This was maintained for
18 hours, upon which the plates were removed and allowed to cool to
room temperature (2 hours). The solvent was removed using a GENEVAC
(45.degree. C., 0.15 mbar) over 5.5 hours. A solution of potassium
t-butoxide (268 mg, 240 .mu.mol) in isopropylalcohol (0.5 ml) was
added to each well, and the plates were sealed and transferred to
an oven at 110.degree. C. under nitrogen. This was maintained for
15 hours, upon which the plates were removed and allowed to cool to
room temperature (2 hours). The solvent was again removed using the
GENEVAC (45.degree. C., 0.15 mbar) over 5.5 hours, and a solution
of p-toluenesulfonic acid (30 mg, 160 .mu.l) in isopropylalcohol
(0.5 ml) was added to each well. The plates were left to stand at
room temperature for 18 hours, and the solvent was removed using
the GENEVAC (45.degree. C., 0.15 mbar) over 5.5 hours. The residues
were dissolved in dimethylsulfoxide (450 .mu.l per well) and each
compound was purified by preparative HPLC. The compounds were
characterised by LC-MS analysis.
Preparative HPLC Conditions
[0186] Column: Phenomenex Luna C18, 5 .mu.m, 150.times.10 mm id
[0187] Temperature: Ambient
[0188] Eluent A: 0.05% Diethylamine (aqueous)
[0189] Eluent B: Acetonitrile
[0190] Sample solvent: 90% dimethylsulfoxide in water
[0191] Initial pump conditions: A % 90, B % 10, flow 6
ml/minute
[0192] Detection: Gilston 119 uv detector -225 nm
[0193] Injection volume -600 .mu.l
3 Gradient Timetable Time (min) A % B % Flow (ml/min) 0.0 95 5 6
0.2 95 5 6 7.0 5 95 6 9.0 5 95 6 9.1 95 5 6 10.5 95 5 6
LC-MS Conditions
[0194] Column: Phenomenex Luna C18, 5 .mu.m, 30.times.4.6 mm
id.
[0195] Temperature: 40.degree. C.
[0196] Eluent A: 0.05% Diethylamine (aqueous)
[0197] Eluent B: Acetonitrile
[0198] Initial pump conditions: A % 90, B % 10, flow 3
ml/minute
[0199] Injection volume -5 .mu.l
[0200] Detection: Start range 210 nm, End range 280 nm, Range
interval 5 nm, threshold 0.1 mAU, peakwidth 0.4 min.
4 Gradient Timetable Time (min) A % B % Flow (ml/min) Pressure
(bar) 0.0 90 10 3 400 2.2 5 95 3 400 2.4 5 95 3 400 2.5 90 10 3
400
[0201] ELSD: Sedere Dedex 55, Temperature: 40.degree. C., Gas Flow:
2.3 bar
[0202] MS: Platform LC, ES+ Cone voltage: 26 v, Capillary: 4.08
kv
[0203] ES- Cone voltage: -24 v, Capillary: -3.58 kv
[0204] Blanket gas: 500 l/min, Temperature: 130.degree. C.
5 Molecular Retention Time Example No. Compound Weight (min.) 1
55-(3-Chloro-benzyl)-3-- isopropyl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 302.1 1.95 2 65-(3-Ethoxy-benzyl)-3-isopropyl-
1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 312.2 1.85 3
75-Cyclohexylmethyl-3-isop- ropyl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 274.2 1.92 4 83-Isopropyl-5-(3-phenoxy-benzyl)-
1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 360.2 2.02 5
93-Isopropyl-5-(2-trifluor- omethyl-
benzyl)-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 302.1 1.85 6
105-(3-Chloro-benzyl)-3-isopropyl- 1,6-dihydro-pyrazolo[4,- 3-
d]pyrimidin-7-one 336.1 1.96 7 115-(4-Chloro-benzyl)-3-- isopropyl-
1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 302.1 1.91 8
125-(4-Benzyloxy-benzyl)-3-isopropyl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 374.2 2.05 9 135-Biphenyl-4-ylmethyl-3--
isopropyl- 1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 344.2 2.04
10 145-[2-(2-Chloro-phenyl)-ethyl]-3- isopropyl-1,6-dihydro-pyrazo-
lo[4,3- d]pyrimidin-7-one 316.1 1.93 11
153-Isopropyl-5-(2,4,6-trifluoro- benzyl)-1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 322.1 1.89 12 165-(3,5-Bis-trifluoromet-
hyl-benzyl)- 3-isopropyl-1,6-dihydro-
pyrazolo[4,3-d]pyrimidin-7-one 404.1 2.07 13
173-Isopropyl-5-(3-trifluoromethoxy-
benzyl)-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 352.1 1.97 14
185-(4-Butoxy-benzyl)-3-isopropyl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 340.2 2.08 15 193-Isopropyl-5-(3-methyl-
-butyl)-1,6- dihydro-pyrazolo[4,3-d]pyrimidin-7- one 248.2 1.80 16
203-Isopropyl-5-(4-methylsulfanyl- benzyl)-1,6-dihydro-pyrazolo[4-
,3- d]pyrimidin-7-one 314.2 1.83 17
215-Ethyl-3-isopropyl-1,6-dihydro- pyrazolo[4,3-d]pyrimidin-7-one
206.1 1.42 18 225-Isobutyl-3-isopropyl-1,6-dihydro-
pyrazolo[4,3-d]pyrimidin-7-one 234.2 1.63 19
233-Isopropyl-5-(4-methoxy-benzyl)- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 298.1 1.77 20 245-(2,5-Dimethyl-benzyl)- -3-
isopropyl-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 296.2 1.93 21
255-Benzhydryl-3-isopropyl-1,6- dihydro-pyrazolo[4,3-d]pyrim-
idin-7- one 344.2 2.07 22 263-Isopropyl-5-(9H-xanthen-9-yl- )-
1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 358.1 2.08 23
275-(2-Fluoro-3-trifluoromethyl- benzyl)-3-isopropyl-1,6-dihydro-
pyrazolo[4,3-d]pyrimidin-7-one 354.1 1.95 24
285-(2,4-Difluoro-benzyl)-3-isopropyl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 304.1 1.82 25 295-(2,3-Difluoro-benzyl)-
-3-isopropyl- 1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 304.1
1.86 26 305-(4-Fluoro-benzyl)-3-isopropyl-
1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 286.1 1.80 27
315-[2-(2-Imidazol-1-yl-e- thoxy)- benzyl]-3-isopropyl-1,6-dihydro-
pyrazolo[4,3-d]pyrimidin-7-one 378.2 1.44 28
325-(5-Fluoro-2-trifluoromethyl- benzyl)-3-isopropyl-1,6-dihydro-
pyrazolo[4,3-d]pyrimidin-7-one 354.1 1.98 29
335-(2,6-Dichloro-benzyl)-3-isopropyl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 336.1 1.97 30 345-(2-Chloro-6-methyl-benzyl)-3-
isopropyl-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 332.1 2.03 31
353-Isopropyl-5-(2-methox- y-benzyl)- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 298.1 1.76 32 365-Cyclopentyl-3-isopropyl-1,6-
dihydro-pyrazolo[4,3-d]pyrimid- in-7- one 246.2 1.83 33
375-(3,4-Difluoro-benzyl)-3-isopro- pyl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 304.1 1.83 34 38N-[1R-1-(3-Isopropyl-7-oxo-6,7-
dihydro-1H-pyrazolo[4,3- d]pyrimidin-5-yl)-2-phenyl-ethyl]-
acetamide 339.2 1.73 35 395-(4-Methyl-benzyl)-3-isopropyl-
1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 282.2 1.83 36
403-Isopropyl-5-[2-(4-met- hoxy- phenyl)-1-phenyl-ethyl]-1,6-
dihydro-pyrazolo[4,3-d]pyrimidin-7- one 388.2 2.13 37
413-Isopropyl-5-napthalen-1- ylmethyl-1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 318.2 2.05 38 425-Cyclopentylmethyl-3-isopropyl-
1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 260.2 1.77 39
435-(2,6-Difluoro-benzyl)- -3-isopropyl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 304.1 1.83 40 445-(3-Methyl-benzyl)-3-isopropyl-
1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 282.2 1.84 41
455-(2,4-Dimethyl-benzyl)- -3- isopropyl-1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 296.2 1.99 42 465-(3-Fluoro-benzyl)-3-isopropyl-
1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 286.1 1.82 43
475-(2,3,6-Trifluoro-benz- yl)-3-
isopropyl-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 322.1 1.91 44
485-(4-Chloro-phenoxymethyl)-3- isopropyl-1,6-dihydro-pyr-
azolo[4,3- d]pyrimidin-7-one 318.1 1.97 45
493-Isopropyl-5-(4-phenoxy-benzyl)- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 360.2 2.05 46 505-(2-Chloro-6-fluoro-be- nzyl)-3-
isopropyl-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 320.1 1.88 47
515-(2-Benzyloxy-benzyl)-3-isopropyl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 374.2 2.04 48 523-Isopropyl-5-(4-methyl-
cyclohexylmethyl)-1,6-dihydro- pyrazolo[4,3-d]pyrimidin-7-one 288.2
2.02 49 533-Isopropyl-5-(1R-1-phenyl-propyl)-
1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 296.2 2.09 50
545-{2-[3-(4-Chloro-pheny- l)- [1,2,4]oxadiazol-5-yl]-ethyl}-3-
isopropyl-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 384.1 2.05 51
553-Isopropyl-5-pentyl-1,- 6-dihydro-
pyrazolo[4,3-d]pyrimidin-7-one 248.2 1.83 52
563-Isopropyl-5-(2-phenoxy-benzyl)- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 360.2 2.03 53 575-(3,5-Dimethyl-benzyl)- -3-
isopropyl-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 296.2 1.97 54
585-(4-Cyclopentyloxy-3-methoxy- benzyl)-3-isopropyl-1,6-dih- ydro-
pyrazolo[4,3-d]pyrimidin-7-one 382.2 1.95 55
593-Isopropyl-5-napthalen-2- ylmethyl-1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 318.2 1.97 56 605-(2,5-Dichloro-
phenylsulfanylmethyl)-3-isopropyl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 368.0 2.09 57 613-Isopropyl-5-(1S-1-phe-
nyl-ethyl)- 1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 282.2 1.95
58 623-Isopropyl-5-(2-methyl-butyl)-1,6- dihydro-pyrazolo[4,3-d]p-
yrimidin-7- one 248.2 1.78 59 635-(2,5-Difluoro-benzyl)-3--
isopropyl- 1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 304.1 1.86
60 645-Benzyl-3-isopropyl-1,6-dihydro- pyrazolo[4,3-d]pyrimidin-7--
one 268.1 1.77 61 653-Isopropyl-5-(4-methyl-pentyl)-
1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 262.2 1.94 62
665-(2-Cyclohexyl-ethyl)-3-isopropyl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 288.2 2.07 63 675-(2-Chloro-4-fluoro-be- nzyl)-3-
isopropyl-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 320.1 1.92 64
683-Isopropyl-5-[2-(4-trifluoromethyl- phenyl)-ethyl]-1,6-dihydro-
pyrazolo[4,3-d]pyrimidin-7-one 350.1 2.03 65
695-(2-Ethoxy-benzyl)-3-isopropyl- 1,6-dihydro-pyrazolo[4,- 3-
d]pyrimidin-7-one 312.2 1.88 66 703-Isopropyl-5-phenoxy-
methyl-1,6- dihydro-pyrazolo[4,3-d]pyrimidin-7- one 284.1 1.85 67
715-(3-Methoxy-benzyl)-3-isopropyl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 298.1 1.76 68 725-(3-Trifluoromethyl-be- nzyl)-3-
isopropyl-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 336.1 1.99 69
735-(4-Isopropyl-benzyl)-3-isopropyl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 310.2 2.04 70
745-(3,5-Difluoro-benzyl)-3-isopropyl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 304.1 1.89 71 755-(2,5-Dimethoxy-benzyl- )-3-
isopropyl-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 328.2 1.80 72
765-(2,3-Dimethyl-benzyl)-3- isopropyl-1,6-dihydro-pyrazolo- [4,3-
d]pyrimidin-7-one 312.2 2.04 73
775-(3,4-Dichloro-benzyl)-3-isopropyl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 336.1 2.02 74 785-(4-Trifluoromethyl-be- nzyl)-3-
isopropyl-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 336.1 1.93 75
795-(2-Methyl-benzyl)-3-isopropyl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 282.2 1.85 76 805-(2-Fluoro-benzyl)-3-isopropyl-
1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 286.1 1.80 77
813-Isopropyl-5-(4-phenyl- -butyl)-1,6-
dihydro-pyrazolo[4,3-d]pyrimidin-7- one 310.2 1.98
[0205] General Procedure for the Preparation of Examples 78 to 159
82
[0206] The carboxylic acids (80 .mu.mol) were dissolved in a 3.75%
solution of triethylamine in dimethylacetamide (400 .mu.l) and
administered to a 96 well plate. Carbonyldiimidazole (13 mg, 80
.mu.mol) dissolved in pyridine (212 .mu.l) was then added into each
well, and the plates were left to stand at room temperature for 2
hours. A solution of 5-substituted-4-amino-pyrazole3-carboxamide
(80 .mu.mol) dissolved in dimethylacetamide (100 .mu.l) was then
added, and the plates were sealed and heated to 70.degree. C. in an
oven under nitrogen. This was maintained for 18 hours, upon which
the plates were removed and allowed to cool to room temperature (2
hours). The solvent was removed using a GENEVAC (30.degree. C.,
0.15 mbar) over 11 hours. A solution of potassium t-butoxide (268
mg, 240 .mu.mol) in isopropylalcohol (0.5 ml) was added to each
well, and the plates were sealed and transferred to an oven at
110.degree. C. under nitrogen. This was maintained for 15 hours,
upon which the plates were removed and allowed to cool to room
temperature (2 hours). The solvent was again removed using the
GENEVAC (30.degree. C., 0.15 mbar) over 11 hours, and a solution of
p-toluenesulfonic acid (30 mg, 160 .mu.l) in isopropylalcohol (0.5
ml) was added to each well. The plates were left to stand at room
temperature for 18 hours, and the solvent was removed using the
GENEVAC (30.degree. C., 0.15 mbar) over 11 hours. The residues were
dissolved in dimethylsulfoxide (450 .mu.l per well) and each
compound was purified by preparative HPLC. The compounds were
characterised by LC-MS analysis.
Preparative HPLC Conditions
[0207] Column: Phenomenex Luna C18, 5 .mu.m, 150.times.10 mm id
[0208] Temperature: Ambient
[0209] Eluent A: 0.05% Diethylamine (aqueous)
[0210] Eluent B: Acetonitrile
[0211] Sample solvent: 90% dimethylsulfoxide in water
[0212] Initial pump conditions: A % 90, B % 10, flow 6
ml/minute
[0213] Detection: Gilston 119 uv detector -225 nm
[0214] Injection volume -600 .mu.l
6 Gradient Timetable Time (min) A % B % Flow (ml/min) 0.0 95 5 6
0.2 95 5 6 7.0 5 95 6 9.0 5 95 6 9.1 95 5 6 10.5 95 5 6
LC-MS Conditions
[0215] Column: Phenomenex Luna C18, 5 .mu.m, 30.times.4.6 mm
id.
[0216] Temperature: 40.degree. C.
[0217] Eluent A: 0.05% Diethylamine (aqueous)
[0218] Eluent B: Acetonitrile
[0219] Initial pump conditions: A % 90, B % 10, flow 3
ml/minute
[0220] Injection volume -5 .mu.l
[0221] Detection: Start range 210 nm, End range 280 nm, Range
interval 5 nm, threshold 0.1 mAU, peakwidth 0.4 min.
7 Gradient Timetable Time (min) A % B % Flow (ml/min) Pressure
(bar) 0.0 90 10 3 400 2.2 5 95 3 400 2.4 5 95 3 400 2.5 90 10 3
400
[0222] ELSD: Sedere Dedex 55, Temperature: 40.degree. C., Gas Flow:
2.3 bar
[0223] MS: Platform LC, ES+ Cone voltage: 26 v, Capillary: 4.08
kv
[0224] ES- Cone voltage: -24 v, Capillary: -3.58 kv
[0225] Blanket gas: 500 l/min, Temperature: 130.degree. C.
8 78 835-(2,6-Dichloro-benzyl)-3-pyridin-3-
yl-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 371.0 1.24 79
845-Cyclopropylmethyl-3-pyridin-3-yl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 267.1 0.92 80 855-(2,6-Difluoro-benzyl)-
-3-pyridin-3- yl-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 339.1
1.10 81 865-(4-Methyl-cyclohexylmethyl)-3-
pyridin-3-yl-1,6-dihydro- pyrazolo[4,3-d]pyrimidin-7-one 323.2 1.46
82 875-(3-Chloro-benzyl)-3-pyridin-3-yl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 337.1 1.23 83
885-(2-Ethoxy-benzyl)-3-pyridin-3-yl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 347.1 1.24 84 895-(2-Phenoxy-benzyl)-3--
pyridin-3- yl-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 395.1
1.46 85 905-(2,3,5-Trifluoro-benzyl)-3-pyridin-
3-yl-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 357.1 1.18 86
915-(3-Fluoro-4-trifluoromethyl-
benzyl)-3-pyridin-3-yl-1,6-dihydro- pyrazolo[4,3-d]pyrimidin-7-one
389.1 1.37 87 925-(5-Fluoro-2-trifluoromethyl-
benzyl)-3-pyridin-3-yl-1,6-dihydro- pyrazolo[4,3-d]pyrimidin-7-one
389.1 1.29 88 935-(5-Bromo-2-methoxy-benzyl)-3-
pyridin-3-yl-1,6-dihydro- pyrazolo[4,3-d]pyrimidin-7-one 411.0 1.30
89 945-(2-Benzyloxy-benzyl)-3-pyridin-3-
yl-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 409.2 1.42 90
953-Butyl-5-(2-methyl-ben- zyl)-1,6-
dihydro-pyrazolo[4,3-d]pyrimidin-7- one 296.2 1.44 91
963-Butyl-5-(2-methoxy-benzyl)-1,6-
dihydro-pyrazolo[4,3-d]pyrimidin- -7- one 312.2 1.36 92
973-Butyl-5-(2-chloro-benzyl)-1,6-
dihydro-pyrazolo[4,3-d]pyrimidin-7- one 316.1 1.45 93
983-Butyl-5-(2-fluoro-benzyl)-1,6-
dihydro-pyrazolo[4,3-d]pyrimidin-7- one 300.14 1.34 94
993-Butyl-5-(2-chloro-6-fluoro- benzyl)-1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 334.1 1.47 95
1003-Butyl-5-(2,6-dichloro-benzyl)-1,6- dihydro-pyrazolo[4,3-d]py-
rimidin-7- one 350.1 1.56 96 1015-Butoxy-3-butyl-1,6-dihyd- ro-
pyrazolo[4,3-d]pyrimidin-7-one 354.2 1.72 97
1023-Butyl-5-cyclopropylmethyl-1,6-
dihydro-pyrazolo[4,3-d]pyrimidin-7- one 246.2 1.14 98
1033-Butyl-5-(2,6-difluoro-benzyl)-1,6-
dihydro-pyrazolo[4,3-d]pyrimidin-7- one 318.1 1.38 99
1043-Butyl-5-(2-ethoxy-benzyl)-1,6-
dihydro-pyrazolo[4,3-d]pyrimidin-7- one 326.2 1.48 100
1053-Butyl-5-(2-benzyloxy-benzyl)-1,6-
dihydro-pyrazolo[4,3-d]pyrimidin-7- one 388.2 1.67 101
1065-(2,4,5-Trifluoro-benzyl)-3-
isopropyl-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 322.1 1.36
102 1075-(2,4-Dichloro-benzy- l)-3-isopropyl-
1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 336.1 1.54 103
1085-(5-Bromo-2-methoxy-benzyl)-3-
isopropyl-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 376.1 1.46
104 1095-(2,3,6-Trichloro-benzyl)-3- isopropyl-1,6-dihydro-pyrazol-
o[4,3- d]pyrimidin-7-one 370.0 1.59 105
1105-(3-Benzyloxy-benzyl)-3-isopropyl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 374.2 1.59 106 1113-isopropyl-5-propyl--
1,6-dihydro- pyrazolo[4,3-d]pyrimidin-7-one 220.1 0.96 107
1125-(2-Trifluoromethoxy-benzyl)-3-
isopropyl-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 352.1 1.49
108 1133-tert-Butyl-5-(3-chl- oro-benzyl)-1,6-
dihydro-pyrazolo[4,3-d]pyrimidin-7- one 316.1 1.61 109
1143-tert-Butyl-5-(5-bromo-2-methoxy-
benzyl)-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 390.1 1.68 110
1153-Isobutyl-5-(2,4,5-trifluoro-benzyl)- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 336.1 1.42 111
1163-Isobutyl-5-(2-methyl-benzyl)-1,6-
dihydro-pyrazolo[4,3-d]pyrimid- in-7- one 296.2 1.40 112
1175-Cyclopentylmethyl-3-isobutyl- -1,6-
dihydro-pyrazolo[4,3-d]pyrimidin-7- one 274.2 1.39 113
1183,5-Diisobutyl-1,6-dihydro- pyrazolo[4,3-d]pyrimidin-7-one 248.2
1.20 114 1193-Isobutyl-5-(2-methoxy-benzyl)-
1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 312.2 1.32 115
1203-Isobutyl-5-(2-chloro-benzyl)-1,6-
dihydro-pyrazolo[4,3-d]pyrimid- in-7- one 316.1 1.41 116
1213-Isobutyl-5-(2-fluoro-benzyl)- -1,6-
dihydro-pyrazolo[4,3-d]pyrimidin-7- one 300.1 1.31 117
1223-Isobutyl-5-(2-chloro-6-fluoro-
benzyl)-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 334.1 1.44 118
1233-Isopropyl-5-(2-meth- yl-butyl)-1,6-
dihydro-pyrazolo[4,3-d]pyrimidin-7- one 262.2 1.35 119
1243-Isobutyl-5-(2-trifluoromethyl- benzyl)-1,6-dihydro-pyrazo-
lo[4,3- d]pyrimidin-7-one 350.1 1.53 120
1253-Isobutyl-5-(2,4-dichloro-benzyl)- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 350.1 1.61 121 1263-Isobutyl-5-(2,6-dic-
hloro-benzyl)- 1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 350.1
1.54 122 1275-(4-Butoxy-benzyl)-3-Isopropyl-
1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 354.2 1.69 123
1285-Cyclopropylmethyl-3-isobutyl-1,6-
dihydro-pyrazolo[4,3-d]pyrimid- in-7- one 246.2 1.08 124
1293-Isobutyl-5-(2,6-difluoro-ben- zyl)- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 318.1 1.34 125
1303-Isobutyl-5-(3-chloro-benzyl)-1,6-
dihydro-pyrazolo[4,3-d]pyrim- idin-7- one 316.1 1.44 126
1313-Isobutyl-5-(2-ethoxy-benzy- l)-1,6-
dihydro-pyrazolo[4,3-d]pyrimidin-7- one 326.2 1.44 127
1323-Isobutyl-5-(2-phenoxy-benzyl)- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 374.2 1.65 128 1333-Isobutyl-5-(2,3,5-t-
rifluoro-benzyl)- 1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 336.1
1.43 129 1343-Isobutyl-5-(5-bromo-2-methoxy-
benzyl)-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 390.1 1.53 130
1353-Isobutyl-5-(2-benzyloxy-benzyl)- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 388.2 1.64 131 1365-[2-(2-Imidazol-1-yl-
-ethoxy)- benzyl]-3-isobutyl-1,6-dihydro-
pyrazolo[4,3-d]pyrimidin-7-one 392.2 1.12 132
1373-Isobutyl-5-(2,3,6-trichloro- benzyl)-1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 384.0 1.66 133
1383-Isobutyl-5-(3-benzyloxy-benzyl)- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 388.2 1.64 134 1395-(2,3-Dihydro-benzof- uran-5-
ylmethyl)-3-isobutyl-1,6-dihydro- pyrazolo[4,3-d]pyrimidin-7-one
324.2 1.25 135 1403-Cyclopentyl-5-(2,4,5-trifluoro-
benzyl)-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 348.1 1.51 136
1413-Cyclopentyl-5-(2-methyl-benzyl)- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 308.2 1.52 137 1423-Cyclopentyl-5-isobu- tyl-1,6-
dihydro-pyrazolo[4,3-d]pyrimidin-7- one 260.2 1.33 138
1433-Cyclopentyl-5-(2-methoxy- benzyl)-1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 324.2 1.44 139 1443-Cyclopentyl-5-(2-ch-
loro-benzyl)- 1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 328.1
1.52 140 1453-Cyclopentyl-5-(2-fluoro-benzyl)-
1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 312.1 1.42 141
1463-Cyclopentyl-5-(2-chloro-6-fluoro-
benzyl)-1,6-dihydro-pyrazolo[4- ,3- d]pyrimidin-7-one 346.1 1.53
142 1473-Cyclopentyl-5-(2-methyl-butyl)- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 274.2 1.49 143 1483-Cyclopentyl-5-(2-tr-
ifluoromethyl- benzyl)-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one
362.1 1.62 144 1493-Cyclopentyl-5-(2,4-dichloro-
benzyl)-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 362.1 1.70 145
1503-Cyclopentyl-5-(2,6-dichloro- benzyl)-1,6-dihydro-pyrazolo[4-
,3- d]pyrimidin-7-one 362.1 1.61 146
1515-(4-Butoxy-benzyl)-3-cyclopentyl- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 366.2 1.80 147 1523-Cyclopentyl-5-cyclo-
propylmethyl- 1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 258.2
1.22 148 1533-Cyclopentyl-5-(2,6-difluoro-
benzyl)-1,6-dihydro-pyraz- olo[4,3- d]pyrimidin-7-one 330.1 1.44
149 1543,5-Dicyclopentyl-1,6-dihydro-
pyrazolo[4,3-d]pyrimidin-7-one 272.2 1.52 150
1553-Cyclopentyl-5-(3-chloro-benzyl)- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 328.1 1.55 151 1563-Cyclopentyl-5-(2,5-dimethoxy-
benzyl)-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 354.2 1.41 152
1573-Cyclopentyl-5-(2-et- hoxy-benzyl)- 1,6-dihydro-pyrazolo[4,3-
d]pyrimidin-7-one 338.2 1.55 153 1583-Cyclopentyl-5-(2-phenoxy-
benzyl)-1,6-dihydro-pyrazolo- [4,3- d]pyrimidin-7-one 386.2 1.75
154 1593-Cyclopentyl-5-(3-fluoro-4-
trifluoromethyl-benzyl)-1,6-dihydro- pyrazolo[4,3-d]pyrimidin-7-one
380.1 1.67 155 1603-Cyclopentyl-5-(2-benzyloxy-
benzyl)-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 400.2 1.73 156
1613-Cyclopentyl-5-[2-(2- -imidazo-1-yl-
ethoxy)-benzyl]-1,6-dihydro- pyrazolo[4,3-d]pyrimidin-7-one 404.2
1.20 157 1623-Cyclopentyl-5-(3-benzyloxy-
benzyl)-1,6-dihydro-pyrazolo[4,3- d]pyrimidin-7-one 400.2 1.73 158
1633-Cyclopentyl-5-propyl-1,6-dihydro- pyrazolo[4,3-d]pyrimidin--
7-one 246.2 1.19 159 1643-Cyclopentyl-5-(2,3-dihydro-
benzofuran-5-ylmethyl)-1,6- dihydro-pyrazolo[4,3-d]pyrimidin-7- one
336.2 1.35
EXAMPLE 160
3-Cyclopentyl-5-(2-trifluoromethoxy-benxyl)-1,6-dihydro-pyrazolo[4,3-d]pyr-
imidin-7-one
[0226] 165
[0227]
5-Cyclopentyl-4-[2-(2-trifluoromethoxy-phenyl)-acetylamino]-1H-pyra-
zole-3-carboxylic acid amide (120 mg, 0.303 mmol) and potassium
tert-butoxide (102 mg, 0.909 mmol) were suspended in
isopropylalcohol (5 ml) and the reaction was heated to reflux,
under nitrogen, for 18 hours. The reaction mixture was concentrated
under reduced pressure and the residue was partitioned between
ethyl acetate (20 ml) and water (20 ml). The aqueous phase was
removed, acidified to pH 2 with 2N HCl, and extracted with ethyl
acetate (2.times.15 ml). The combined organic extracts were washed
with saturated sodium carbonate solution (3.times.10 ml), dried
over MgSO.sub.4, concentrated under reduced pressure and the
residue was purified by flash column chromatography on silica gel
eluting with dichloromethane: methanol (95:5, by volume) to give
3-cyclopentyl-5-(2-trifluoromethoxy-benxyl)-1,6-dihydro-pyrazolo[4,3-d]py-
rimidin-7-one (21 mg) as an off-white solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6): .delta.=7.36-7.41 (2H, m), 7.29-7.36 (2H, m),
3.97-4.03 (2H, brs), 2.39-2.45 (1H, m, partially masked by
solvent), 1.82-1.94 (2H, m), 1.66-1.79 (2H, m), 1.58-1.65 (2H, m),
1.49-1.58 (2H, m) ppm. LRMS (electrospray): m/z [M-H]+377.
EXAMPLE 161
3-Isobutyl-5-(2-trifluoromethoxy-benxyl)-116-dihydro-pyrazolo[4,3-d]pyrimi-
din-7-one
[0228] 166
[0229]
5-Isobutyl-4-[2-(2-trifluoromethoxy-phenyl)-acetylamino]-1H-pyrazol-
e-3-carboxylic acid amide (140 mg, 0.365 mmol) and potassium
tert-butoxide (123 mg, 1.09 mmol) were suspended in
isopropylalcohol (6 ml) and the reaction was heated to reflux,
under nitrogen, for 18 hours. The reaction mixture was concentrated
under reduced pressure and the residue was partitioned between
ethyl acetate (20 ml) and water (20 ml). The aqueous phase was
removed, acidified to pH 2 with 2N HCl, and extracted with ethyl
acetate (2.times.15 ml). The combined organic extracts were washed
with saturated sodium carbonate solution (3.times.10 ml), dried
over MgSO.sub.4, concentrated under reduced pressure and the
residue was purified by flash column chromatography on silica gel
eluting with dichloromethane:methanol (95:5, by volume) to give
3-isobutyl-5-(2-trifluoromethoxy-benxyl)-1,6-dihydro-pyrazolo[4,3-d]pyrim-
idin-7-one (27 mg) as an off-white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta.=8.64-8.74 (1H, brs), 7.22-7.41 (4H, m,
partially masked by solvent), 4.15 (2H, s), 2.79-2.84 (2H, d),
2.13-2.23 (1H, m), 0.92-1.00 (6H, d) ppm. LRMS (electrospray): m/z
[M+H].sup.+ 367, [M-H].sup.+ 365.
EXAMPLE 162
3-Pyridin-3-yl-5-(2-trifluoromethoxy-benxyl)-1,6-dihydro-pyrazolo[4,3-d]py-
rimidin-7-one
[0230] 167
[0231]
5-Pyridin-3-yl-4-[2-(2-trifluoromethoxy-phenyl)-acetylamino]-1H-pyr-
azole-3-carboxylic acid amide (345 mg, 0.85 mmol) and potassium
tert-butoxide (286 mg, 2.55 mmol) were suspended in
isopropylalcohol (5 ml) and the reaction was heated to 55.degree.
C. under nitrogen for 18 hours. The reaction mixture was
concentrated under reduced pressure and the residue was partitioned
between ethyl acetate (20 ml) and water (20 ml). The aqueous phase
was removed, acidified to pH 2 with 2N HCl, and extracted with
ethyl acetate (2.times.15 ml) and dichloromethane (2.times.15 ml).
The combined organic extracts were dried over MgSO.sub.4,
concentrated under reduced pressure and the residue was purified by
flash column chromatography on silica gel eluting with a solvent
gradient of dichloromethane:methanol (99:1 changing to 95:5, by
volume). The product was triturated with methanol (3 ml),
dichloromethane (3 ml) and diethylether (3 ml) to give
3-pyridin-3-yl-5-(2-trifluorometho-
xy-benxyl)-1,6-dihydro-pyrazolo[4,3-d]pyrimidin-7-one (13 mg) as an
off-white solid. .sup.1H NMR (400 MHz, CD.sub.3OD): .delta.=9.34
(1H, brs), 8.57-8.61 (1H, d), 8.43-8.48 (1H, m), 7.32-7.47 (5H, m),
4.18 (2H, s) ppm. LRMS (electrospray): m/z [M-H].sup.+ 386.
Preparation 1
4-Methyl-3-oxo-pentanoic Acid Ethyl Ester
[0232] 168
[0233] Sodium pellets (3.39 g, 148 mmol) were dissolved in ethanol
(100 ml) under nitrogen at room temperature and a solution of
diethyloxalate (20 ml, 147 mmol) in 3-methyl-2-butanone (18.9 ml,
177 mmol) was added dropwise at room temperature over 30 minutes.
The reaction was diluted with ethanol (100 ml), heated to
60.degree. C. and stirred at this temperature for 2 hours. After
cooling to room temperature the reaction was poured onto ice-cold
2N HCl (200 ml) and extracted with diethylether (300 ml) and ethyl
acetate (300 ml). The combined organic extracts were dried over
MgSO.sub.4, concentrated under reduced pressure and the residue was
purified by flash column chromatography on silica gel eluting with
a solvent gradient of pentane: ethyl acetate (99:1 changing to
95:5, by volume) to give 4-methyl-3-oxo-pentanoic acid ethyl ester
(23.8 g) as a yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3):
.delta.=14.40-14.80 (1H, brs), 6.40 (1H, s), 4.30-4.39 (2H, quart),
2.60-2.71 (1H, quin), 1.35-1.40 (3H, t), 1.15-1.20 (6H, d) ppm.
LRMS (electrospray): m/z [M-H].sup.+ 185.
Preparation 2
5-Isopropyl-1H-pyrazol-3-carboxylic Acid Ethyl Ester
[0234] 169
[0235] Hydrazine hydrate (6.6 ml, 134 mmol) was added to a solution
of 4-methyl-3-oxo-pentanoic acid ethyl ester (23.8 g, 188 mmol) in
ethanol (100 ml) at room temperature under nitrogen. The reaction
was allowed to proceed at room temperature for 18 hours, and the
solvent was removed under reduced pressure. The residue was
partitioned between dichloromethane (300 ml) and water (300 ml) and
the aqueous phase was removed. The organic phase was washed with
water (2.times.200 ml), dried over MgSO.sub.4 and concentrated
under reduced pressure. The residue was purified by flash column
chromatography on silica gel eluting with a solvent gradient of
pentane:ethyl acetate (4:1 changing to 2:1, by volume) to give
5-isopropyl-1H-pyrazol-3-carboxylic acid ethyl ester (18.9 g) as a
white solid. .sup.1H NMR (400 MHz, CDCl.sub.3): .delta.=10.80-10.95
(1H, brs), 6.61 (1H, s), 4.33-4.40 (2H, quart), 2.98-3.08 (1H,
quin), 1.35-1.41 (3H, t), 1.24-1.32 (6H, d) ppm. LRMS
(electrospray): m/z [M-H].sup.+ 181.
Preparation 3
5-Isopropyl-1H-pyrazol-3-carboxylic Acid
[0236] 170
[0237] 5-Isopropyl-1H-pyrazol-3-carboxylic acid ethyl ester (18.9
g, 104 mmol) and 1M NaOH solution (260 ml, 259 mmol) were dissolved
in 1,4-dioxan (300 ml), the reaction was heated to 50.degree. C.
under nitrogen and stirred for 3 hours. The reaction mixture was
cooled, adjusted to pH 2 using concentrated hydrochloric acid and
the solvent was removed under reduced pressure. The residual solid
was azeotroped with toluene (2.times.30 ml), dissolved in ethyl
acetate (500 ml) and washed with water (200 ml). The aqueous phase
was removed, extracted with ethyl acetate (2.times.200 ml) and the
combined organic extracts were dried over MgSO.sub.4. The solvent
was removed under reduced pressure and the residue was azeotroped
with dichloromethane (2.times.50 ml) to give
5-isopropyl-1H-pyrazol-3-carboxylic acid (14.7 g) as a white solid.
.sup.1H NMR (400 MHz, DMSO-D6): .delta.=12.50-13.30 (2H, brs), 6.42
(1H, s), 2.84-2.94 (1H, quin), 1.15-1.19 (6H, d) ppm. LRMS
(electrospray): m/z [M-H].sup.+ 153.
Preparation 4
5-Isopropyl-4-nitro-1H-pyrazol-3-carboxylic Acid
[0238] 171
[0239] 5-Isopropyl-1H-pyrazol-3-carboxylic acid (5 g, 32.5 mmol)
was added portionwise to concentrated sulfuric acid (25 ml) at room
temperature with stirring. The reaction mixture was then heated to
60.degree. C. and concentrated nitric acid (70%, 6 ml, 90 mmol) was
added dropwise, keeping the temperature at 60.degree. C. The
reaction was then stirred at 60.degree. C. for 3 hours, cooled to
room temperature and poured onto 50 ml of ice with stirring. After
15 minutes the white precipitate was isolated by filtration, washed
with water and dried under reduced pressure to give
5-isopropyl-4-nitro-1H-pyrazol-3-carboxylic acid (5.2 g) as a white
solid. .sup.1H NMR (400 MHz, DMSO-D6): .delta.=13.86-13.93 (1H,
brs), 13.50-13.80 (1H, brs), 3.39-3.52 (1H, m), 1.18-1.30 (6H, d)
ppm. LRMS (electrospray): m/z [M-H].sup.+ 198.
Preparation 5
5-Isopropyl-4-nitro-1H-pyrazol-3-carboxylic Acid Amide
[0240] 172
[0241] Oxalyl chloride (6.8 ml, 77.6 mmol) was added dropwise to a
suspension of 5-isopropyl-4-nitro-1H-pyrazol-3-carboxylic acid
(5.15 g, 25.9 mmol) in dichloromethane (80 ml) containing
dimethylformamide (0.1 ml) under nitrogen at 0.degree. C. The
reaction was stirred at 0.degree. C. for 1 hours, allowed to warm
to room temperature and stirred for a further 2 hours. The solvent
was removed under reduced pressure, the residue was dissolved in
toluene (100 ml) and ammonia gas was bubbled into the solution for
2 hours. The reaction was stirred under nitrogen at room
temperature for 18 hours, concentrated under reduced pressure and
the residue was dissolved in hot methanol (300 ml). The resultant
precipitate was filtered and the filtrate was concentrated under
reduced pressure. The residue was azeotroped with water (300 ml),
concentrated to approximately 80 ml under reduced pressure and the
precipitate was isolated by filtration. This was washed with water
and dried under to give 5-isopropyl-4-nitro-1H-pyrazol-3-carboxylic
acid amide (3.1 g) as an orange solid. .sup.1H NMR (400 MHz,
DMSO-D6): .delta.=7.94-7.99 (1H, brs), 7.68-7.72 (1H, brs),
3.45-3.55 (1H, m), 1.24-1.30 (6H, d) ppm. LRMS (electrospray): m/z
[M+Na].sup.+ 221, [M-H].sup.+ 197.
Preparation 6
4-Amino-5-isopropyl-1H-pyrazol-3-carboxylic Acid Amide
[0242] 173
[0243] 5-Isopropyl-4-nitro-1H-pyrazol-3-carboxylic acid amide (3 g,
15.1 mmol) and 10% palladium on carbon (500 mg) in ethanol (30 ml)
were stirred under hydrogen (50 psi) at room temperature for 18
hours. The reaction mixture was filtered and the solid was washed
with methanol (50 ml), dichloromethane (50 ml), ethanol (50 ml) and
ethyl acetate (50 ml). The filtrate was concentrated under reduced
pressure and the residue was purified by flash column
chromatography on silica gel eluting with dichloromethane:methanol
(9:1, by volume) to give 4-amino-5-isopropyl-1H--
pyrazol-3-carboxylic acid amide (2.6 g) as an off-white solid.
.sup.1H NMR (400 MHz, DMSO-D6): .delta.=12.20-12.30 (1H, brs),
7.02-7.14 (1H, brs), 6.85-6.95 (1H, brs), 4.30-4.46 (2H, brs),
2.90-3.00 (1H, m), 1.15-1.21 (6H, d) ppm. LRMS (electrospray): m/z
[M-H].sup.+ 167, [2M-H].sup.+ 335. Anal. Found C, 49.86; H, 7.21;
N, 33.07. C.sub.7H.sub.12N.sub.4O requires C, 49.99; H, 7.19; N,
33.31%.
Preparation 7
3-Oxo-heptanoic Acid Ethyl Ester
[0244] 174
[0245] Sodium pellets (3.82 g, 166 mmol) were dissolved in ethanol
(100 ml) under nitrogen at room temperature and a solution of
diethyloxalate (22.6 ml, 166 mmol) hexan-2-one (20 g, 198 mmol) was
added dropwise at room temperature over 30 minutes. The reaction
was diluted with ethanol (100 ml), heated to 60.degree. C. and
stirred at this temperature for 2 hours. After cooling to room
temperature the reaction was poured onto ice-cold 2N HCl (200 ml)
and extracted with diethylether (300 ml) and ethyl acetate (300
ml). The combined organic extracts were dried over MgSO.sub.4,
concentrated under reduced pressure and the residue was purified by
flash column chromatography on silica gel eluting with a solvent
gradient of pentane: ethyl acetate (99:1 changing to 95:5, by
volume) to give 3-oxo-heptanoic acid ethyl ester (30.3 g) as an
orange oil. .sup.1H NMR (400 MHz, CDCl.sub.3): .delta.=14.30-14.80
(1H, brs), 6.37 (1H, s), 4.30-4.39 (2H, quart), 2.43-2.50 (2H, t),
1.59-1.62 (2H, quin), 1.31-1.40 (5H, t+m), 0.86-0.97 (3H, t) ppm.
LRMS (electrospray): m/z [M-H]+199.
Preparation 8
5-Butyl-1H-Pyrazol-3-carboxylic Acid Ethyl Ester
[0246] 175
[0247] Hydrazine hydrate (7.75 ml, 157 mmol) was added dropwise to
a solution of 3-oxo-heptanoic acid ethyl ester (30 g, 150 mmol) in
ethanol (100 ml) at room temperature under nitrogen. The reaction
was heated to 50.degree. C. and allowed to proceed at this
temperature for 18 hours, and the solvent was removed under reduced
pressure. The residue was partitioned between dichloromethane (300
ml) and water (300 ml) and the aqueous phase was removed. The
organic phase was washed with water (2.times.200 ml), dried over
MgSO.sub.4 and concentrated under reduced pressure. The residue was
purified by flash column chromatography on silica gel eluting with
pentane: ethyl acetate (8:1, by volume) to give
5-butyl-1H-pyrazol-3-carboxylic acid ethyl ester (24 g) as a yellow
oil. .sup.1H NMR (400 MHz, CDCl.sub.3): .delta.=10.58-10.78 (1H,
brs), 6.62 (1H, s), 4.35-4.40 (2H, quart), 2.63-2.70 (2H, t),
1.60-1.67 (2H, quin), 1.35-1.42 (5H, t+m), 0.90-0.96 (3H, t) ppm.
LRMS (electrospray): m/z [M-H].sup.+ 195.
Preparation 9
5-Butyl-1H-pyrazol-3-carboxylic Acid
[0248] 176
[0249] 5-Butyl-1H-pyrazol-3-carboxylic acid ethyl ester (24 g, 122
mmol) and 1M NaOH solution (305 ml, 306 mmol) were dissolved in
1,4-dioxan (300 ml), the reaction was heated to 55.degree. C. under
nitrogen and stirred for 2 hours. The reaction mixture was cooled,
adjusted to pH 2 using concentrated hydrochloric acid and the
solvent was removed under reduced pressure. The residual solid was
dissolved in ethyl acetate (300 ml) and washed with water (300 ml).
The aqueous phase was removed, extracted with ethyl acetate (300
ml) and the combined organic extracts were dried over MgSO.sub.4.
The solvent was removed under reduced pressure and the residue was
azeotroped with dichloromethane (2.times.50 ml) to give
5-butyl-1H-pyrazol-3-carboxylic acid (22.6 g) as a white solid.
.sup.1H NMR (400 MHz, DMSO-D6): .delta.=12.50-13.00 (2H, brs), 6.41
(1H, s), 2.47-2.57 (2H, t), 1.46-1.56 (2H, quin), 1.19-1.29 (2H,
sext), 1.15-1.19 (3H, t) ppm. LRMS (electrospray): m/z [M-H]+167.
Anal. Found C, 57.01; H, 7.23; N, 16.50.
C.sub.8H.sub.12N.sub.2O.sub.2 requires C, 57.13; H, 7.19; N,
16.66%.
Preparation 10
5-Butyl-4-nitro-1H-pyrazol-3-carboxylic Acid
[0250] 177
[0251] 5-Butyl-1H-pyrazol-3-carboxylic acid (22.6 g, 134 mmol) was
added portionwise to concentrated sulfuric acid (100 ml) at room
temperature with stirring. The reaction mixture was then heated to
60.degree. C. and concentrated nitric acid (70%, 23.7 ml, 376 mmol)
was added dropwise, keeping the temperature at 60.degree. C. The
reaction was then stirred at 60.degree. C. for 3 hours, cooled to
room temperature and poured onto 50 ml of ice with stirring. After
15 minutes the pale yellow precipitate was isolated by filtration,
washed with water and dried under reduced pressure to give
5-butyl-4-nitro-1H-pyrazol-3-carboxylic acid (21.9 g) as a pale
yellow solid. .sup.1H NMR (400 MHz, DMSO-D6): .delta.=2.83-2.92
(2H, t), 1.56-1.64 (2H, quin), 1.22-1.36 (2H, sext), 0.84-0.90 (3H,
t) ppm. LRMS (electrospray): m/z [M-H].sup.+ 212. Anal. Found C,
30.19; H, 5.41; N, 13.12. C.sub.8H.sub.11N.sub.3O.sub.4. 6 mol
H.sub.2O requires C, 29.91; H, 7.22; N, 13.08%.
Preparation 11
5-Butyl-4-nitro-1H-pyrazol-3-carboxylic Acid Amide
[0252] 178
[0253] Oxalyl chloride (12.3 ml, 141 mmol) was added dropwise to a
suspension of 5-butyl-4-nitro-1H-pyrazol-3-carboxylic acid (10 g,
46.9 mmol) in dichloromethane (100 ml) containing dimethylformamide
(0.5 ml) under nitrogen at 0.degree. C. The reaction was stirred at
0.degree. C. for 1 hours, allowed to warm to room temperature and
stirred for a further 2 hours. The solvent was removed under
reduced pressure, the residue was dissolved in toluene (100 ml) and
ammonia gas was bubbled into the solution for 2 hours. The reaction
was stirred under nitrogen at room temperature for 18 hours,
concentrated under reduced pressure and the residue was purified by
flash column chromatography on silica gel eluting with
dichloromethane:methanol (9:1, by volume) to give
5-butyl-4-nitro-1H-pyrazol-3-carboxylic acid amide (3.1 g) as an
orange solid. .sup.1H NMR (400 MHz, DMSO-D6): .delta.=7.87-7.96
(1H, brs), 7.57-7.66 (1H, brs), 2.83-2.90 (2H, t), 1.56-1.63 (2H,
quin), 1.24-1.36 (2H, sext), 0.84-0.92 (3H, t) ppm. LRMS
(electrospray): m/z [M-H].sup.+ 211. Anal. Found C, 44.66; H, 5.56;
N, 25.50. C.sub.8H.sub.12N.sub.4O.sub- .3. 0.23 mol H.sub.2O
requires C, 44.41; H, 5.80; N, 25.90%.
Preparation 12
4-Amino-5-butyl-1H-pyrazol-3-carboxylic Acid Amide
[0254] 179
[0255] 5-butyl-4-nitro-1H-pyrazol-3-carboxylic acid amide (3.1 g,
17.0 mmol) and 10% palladium on carbon (600 mg) in ethanol (50 ml)
were stirred under hydrogen (50 psi) at room temperature for 18
hours. The reaction mixture was filtered and the solid was washed
with methanol (50 ml), dichloromethane (50 ml), ethanol (50 ml) and
ethyl acetate (50 ml). The filtrate was concentrated under reduced
pressure and the residue was purified by flash column
chromatography on silica gel eluting with a solvent gradient of
dichloromethane:methanol (95:5 changing to 90:10, by volume) to
give 4-amino-5-butyl-1H-pyrazol-3-carboxylic acid amide (2.37 g) as
an an orange solid. .sup.1H NMR (400 MHz, DMSO-D6):
.delta.=12.24-12.32 (1H, brs), 7.02-7.14 (1H, brs), 6.80-6.95 (1H,
brs), 4.28-4.46 (2H, brs), 2.39-2.50 (2H, t, partially masked by
solvent), 1.45-1.56 (2H, quin), 1.22-1.35 (2H, sext), 0.83-0.90
ppm. LRMS (electrospray): m/z [M-H].sup.+ 181, [2M-H].sup.+ 363.
Anal. Found C, 52.58; H, 7.80; N, 30.56. C.sub.8H.sub.14N.sub.4O
requires C, 52.73; H, 7.74; N, 30.75%.
Preparation 13
4,4-Dimethyl-3-oxo-pentanoic Acid Ethyl Ester
[0256] 180
[0257] Sodium pellets (4.6 g, 200 mmol) were dissolved in ethanol
(165 ml) under nitrogen at room temperature and a solution of
diethyloxalate (27.2 ml, 200 mmol) in tert-butyl-methyl ketone
(20.1 g, 200 mmol) was added dropwise at room temperature over 15
minutes. The reaction was diluted with ethanol (100 ml), heated to
60.degree. C. and stirred at this temperature for 2 hours. After
cooling to room temperature the reaction was stirred for 64 hours,
poured onto ice-cold 2N HCl (200 ml) and extracted with
diethylether (3.times.200 ml). The combined organic extracts were
washed with water, dried over MgSO.sub.4 and concentrated under
reduced pressure to give 4,4-dimethyl-3-oxo-pentanoic acid ethyl
ester (36.7 g) as a yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3):
.delta.=6.48 (1H, s), 4.26-4.37 (2H, quart), 1.29-1.38 (3H, t),
1.17 (9H, s) ppm. LRMS (electrospray): m/z [M+Na].sup.+ 223,
[M-H]+199.
Preparation 14
5-tert-Butyl-1H-pyrazol-3-carboxylic Acid Ethyl Ester
[0258] 181
[0259] Hydrazine hydrate (9.5 ml, 180 mmol) was added to a solution
of 4,4-dimethyl-3-oxo-pentanoic acid ethyl ester (36.7 g, 180 mmol)
in ethanol (188 ml) at room temperature under nitrogen. The
reaction was allowed to proceed at room temperature for 2 hours,
and the solvent was removed under reduced pressure. The residue was
partitioned between dichloromethane (500 ml) and water (400 ml) and
the aqueous phase was removed. The organic phase was washed with
brine (200 ml), dried over MgSO.sub.4 and concentrated under
reduced pressure to give 5-tert-butyl-1H-pyrazol-3-carboxylic acid
ethyl ester (30.6 g) as a white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta.=14.50-14.90 (1H, brs), 6.45 (1H, s), 4.25-4.31
(2H, quart), 1.27-1.36 (3H, t), 1.16 (9H, s) ppm. LRMS
(thermospray): m/z [M+H].sup.+ 197. Anal. Found C, 61.12; H, 8.20;
N, 14.28. C.sub.10H.sub.16N.sub.2O.sub.2 requires C, 61.20; H,
8.22; N, 14.27%.
Preparation 15
5-tert-Butyl-1H-pyrazol-3-carboxylic Acid
[0260] 182
[0261] 5-tert-Butyl-1H-pyrazol-3-carboxylic acid ethyl ester (20 g,
100 mmol) and 1M NaOH solution (250 ml, 250 mmol) were dissolved in
1,4-dioxan (300 ml), the reaction was heated to 60.degree. C. under
nitrogen and stirred for 2.5 hours. The reaction was then cooled to
room temperature and stirred for a further 18 hours. The reaction
mixture was adjusted to pH 2 using concentrated hydrochloric acid,
extracted with ethyl acetate (4.times.200 ml) and the combined
organic extracts washed with brine (100 ml). The organic phase was
dried over MgSO.sub.4 and the solvent was removed under reduced
pressure to give 5-tert-butyl-1H-pyrazol-3-carboxylic acid (14.7 g)
as a pale yellow solid. .sup.1H NMR (400 MHz, DMSO-D6):
.delta.=12.80-12.88 (2H, brs), 6.41 (1H, s), 1.11 (9H, s) ppm. LRMS
(electrospray): m/z [M+H].sup.+ 169, [M+Na].sup.+ 191, [M-H].sup.+
167.
Preparation 16
5-tert-Butyl-4-nitro-1H-pyrazol-3-carboxylic Acid
[0262] 183
[0263] 5-tert-Butyl-1H-pyrazol-3-carboxylic acid (5 g, 29.7 mmol)
was added portionwise to concentrated sulfuric acid (25 ml) at room
temperature with stirring. The reaction mixture was then heated to
60.degree. C. and concentrated nitric acid (70%, 5.15 ml) was added
dropwise, keeping the temperature at 60.degree. C. The reaction was
then stirred at 60.degree. C. for 2.5 hours, cooled to room
temperature and poured onto 50 ml of ice with stirring. After 15
minutes the white precipitate was isolated by filtration, washed
with water and dried under reduced pressure to give
5-tert-butyl-4-nitro-1H-pyrazol-3-carboxylic acid (6.0 g) as a
white solid. .sup.1H NMR (400 MHz, DMSO-D6): .delta.=13.50-13.88
(2H, brs), 1.13 (9H, s) ppm. LRMS (electrospray): m/z [M+Na].sup.+
236, [M-H].sup.+ 212.
Preparation 17
5-tert-Butyl-4-nitro-1H-pyrazol-3-carboxylic Acid Amide
[0264] 184
[0265] Oxalyl chloride (10.2 ml, 117 mmol) was added dropwise to a
suspension of 5-tert-butyl-4-nitro-1H-pyrazol-3-carboxylic acid (6
g, 28 mmol) in dichloromethane (55 ml) containing dimethylformamide
(0.1 ml) under nitrogen at 0.degree. C. The reaction was stirred at
0.degree. C. for 0.5 hours, allowed to warm to room temperature and
stirred for a further 1.5 hours. The solvent was removed under
reduced pressure, the residue was azeotroped with dichloromethane
(50 ml) and the residue was dissolved in dichloromethane (100 ml).
Ammonia gas was bubbled into the solution for 45 minutes and the
reaction was stirred under nitrogen at room temperature for 18
hours, concentrated under reduced pressure and the residue was
dissolved in ethyl acetate (250 ml). After washing with water (100
ml) and brine (100 ml) the organic phase was filtered, the filtrate
was dried over MgSO.sub.4 and concentrated under reduced pressure
to give 5-tert-butyl-4-nitro-1H-pyrazol-3-carboxylic acid amide
(4.0 g) as a light brown solid. .sup.1H NMR (400 MHz, DMSO-D6):
.delta.=12.90-13.08 (1H, brs), 7.78-7.86 (1H, brs), 7.49-7.60 (1H,
brs), 1.30 (9H, s) ppm. LRMS (electrospray): m/z [M+H].sup.+ 213,
[M+Na].sup.+ 235, [M-H].sup.+ 211.
Preparation 18
4-Amino-5-tert-butyl-1H-pyrazol-3-carboxylic Acid Amide
[0266] 185
[0267] 5-tert-Butyl-4-nitro-1H-pyrazol-3-carboxylic acid amide (4.6
g, 21 mmol) and 10% palladium on carbon (300 mg) in ethanol (80 ml)
was stirred under hydrogen (60 psi) at room temperature for 18
hours. The reaction mixture was filtered and the filtrate was
concentrated under reduced pressure. The residue was pre-absorbed
onto silica gel and purified by flash column chromatography eluting
with a solvent gradient of dichloromethane: methanol (100:0
changing to 95:5 then 90:10, by volume) to give
4-amino-5-tert-butyl-1H-pyrazol-3-carboxylic acid amide (2.96 g) as
an off-white solid, which was a mixture of rotamers. .sup.1H NMR
(400 MHz, DMSO-D6): .delta.=12.10-12.20 (0.75H, brs), 11.75-11.85
(0.25H, brs), 7.04-7.16 (1.5H, brs), 6.88-6.96 (0.5H, brs),
4.27-4.59 (2H, 2xbrs), 1.12 (9H, s) ppm. LRMS (electrospray): m/z
[M+H].sup.+ 183, [M+Na].sup.+ 205, [M-H]+181. Anal. Found C, 52.45;
H, 7.84; N, 30.62. C.sub.8H.sub.14N.sub.4O requires C, 52.73; H,
7.74; N, 30.75%.
Preparation 19
5-Methyl-3-oxo-hexanoic Acid Ethyl Ester
[0268] 186
[0269] Sodium pellets (4.6 g, 200 mmol) were dissolved in ethanol
(165 ml) under nitrogen at room temperature and a solution of
diethyloxalate (13.5 ml, 100 mmol) in isobutylmethyl ketone (30 ml,
200 mmol) was added dropwise at room temperature over 20 minutes.
The reaction was heated to 60.degree. C. and stirred at this
temperature for one hour. After cooling to room temperature the
reaction was poured onto ice-cold 2N HCl (200 ml) and extracted
with diethylether (4.times.200 ml). The combined organic extracts
were washed with water, dried over MgSO.sub.4 and concentrated
under reduced pressure to give 5-methyl-3-oxo-hexanoic acid ethyl
ester (20 g) as a yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3):
.delta.=14.40-14.70 (1H, brs), 6.27 (1H, s), 4.25-4.32 (2H, quart),
2.26-2.31 (2H, d), 2.02-2.18 (1H, m), 1.29-1.34 (3H, t), 0.89-0.94
(6H, d) ppm. LRMS (thermospray): m/z [M+NH.sub.4].sup.+ 218.
Preparation 20
5-Isobutyl-1H-pyrazol-3-carboxylic Acid Ethyl Ester
[0270] 187
[0271] Hydrazine hydrate (5.7 ml, 115 mmol) was added to a solution
of 5-methyl-3-oxo-hexanoic acid ethyl ester (22 g, 110 mmol) in
ethanol (113 ml) at room temperature under nitrogen. The reaction
was allowed to proceed at room temperature for 18 hours, and the
solvent was removed under reduced pressure. The residue was
partitioned between dichloromethane (400 ml) and water (400 ml) and
the aqueous phase was removed. The organic phase was washed with
brine (200 ml), water (200 ml), dried over MgSO.sub.4 and
concentrated under reduced pressure. The residue was purified by
flash column chromatography on silica gel eluting with a solvent
gradient of pentane:ethyl acetate (1:0 changing to 6:1, 5:1, 4:1,
3:1, 2:1 and finally 1:1, by volume) to give
5-isobutyl-1H-pyrazol-3-carboxylic acid ethyl ester (16.5 g) as a
white solid. .sup.1H NMR (400 MHz, CDCl.sub.3): .delta.=11.60-12.60
(1H, brs), 6.53 (1H, s), 4.26-4.35 (2H, quart), 2.48-2.54 (2H, d),
1.80-1.90 (1H, m), 1.25-1.31 (3H, t), 0.81-0.88 (6H, d) ppm. LRMS
(thermospray): m/z [M+H].sup.+ 197, [2M+H].sup.+ 393. Anal. Found
C, 61.49; H, 8.30; N, 14.24. C.sub.10H.sub.16N.sub.2O.sub.2
requires C, 61.20; H, 8.22; N, 14.27%.
Preparation 21
5-Isobutyl-1H-pyrazol-3-carboxylic Acid
[0272] 188
[0273] 5-Isobutyl-1H-pyrazol-3-carboxylic acid ethyl ester (16.2 g,
83 mmol) and 1M NaOH solution (173 ml, 173 mmol) were dissolved in
1,4-dioxan (260 ml) and the reaction was stirred at room
temperature under nitrogen for 64 hours. The reaction mixture was
adjusted to pH 7 using concentrated hydrochloric acid, and
concentrated under reduced pressure. Water (500 ml) was added, the
slurry was adjusted to pH 1 with concentrated hydrochloric acid and
the aqueous phase was extracted with ethyl acetate (5.times.300
ml). The combined organic extracts were dried over MgSO.sub.4 and
the solvent was removed under reduced pressure to give
5-isobutyl-1H-pyrazol-3-carboxylic acid (10 g) as a white solid.
.sup.1H NMR (400 MHz, DMSO-D6): .delta.=12.72-12.90 (1H, brs), 6.39
(1H, s), 2.39-2.43 (2H, d), 1.77-1.86 (1H, m), 0.78-0.83 (6H, d)
ppm. LRMS (electrospray): m/z [M+Na].sup.+ 191, [2M+Na].sup.+ 359,
[M-H].sup.+ 167, [2M-H].sup.+ 335.
Preparation 22
5-Isobutyl-4-nitro-1H-pyrazol-3-carboxylic Acid
[0274] 189
[0275] 5-Isobutyl-1H-pyrazol-3-carboxylic acid (5 g, 29.7 mmol) was
added portionwise to concentrated sulfuric acid (25 ml) at room
temperature with stirring. The reaction mixture was then heated to
60.degree. C. and concentrated nitric acid (70%, 5.15 ml) was added
dropwise, keeping the temperature at 60.degree. C. The reaction was
then stirred at 60.degree. C. for 3 hours, cooled to room
temperature and poured onto 50 ml of ice with stirring. The
resultant white precipitate was isolated by filtration, washed with
water and dried under reduced pressure to give
5-isobutyl-4-nitro-1H-pyrazol-3-carboxylic acid (6.4 g) as a white
solid. .sup.1H NMR (400 MHz, DMSO-D6): .delta.=2.71-2.76 (2H, d),
1.88-2.00 (1H, m), 0.80-0.87 (6H, d) ppm. LRMS (thermospray): m/z
[M+NH.sub.4].sup.+ 231, [M-H].sup.+ 212. LRMS (electrospray): m/z
[M-H].sup.+ 212, [2M-H].sup.+ 425. Anal. Found C, 42.54; H, 5.18;
N, 18.63. C.sub.8H.sub.11N.sub.3O.sub.4. 0.7 mol H.sub.2O requires
C, 42.55; H, 5.54; N, 18.61%.
Preparation 23
5-Isobutyl-4-nitro-1H-pyrazol-3-carboxylic Acid Amide
[0276] 190
[0277] Oxalyl chloride (10 ml, 115 mmol) was added dropwise to a
suspension of 5-isobutyl-4-nitro-1H-pyrazol-3-carboxylic acid (5.6
g, 26 mmol) in dichloromethane (70 ml) containing dimethylformamide
(0.1 ml) under nitrogen at 0.degree. C. The reaction was stirred at
0.degree. C. for 0.5 hours, allowed to warm to room temperature and
stirred for a further 2 hours. The solvent was removed under
reduced pressure, the residue was azeotroped with dichloromethane
(3.times.50 ml) and the residue was dissolved in toluene (100 ml).
Ammonia gas was bubbled into the solution for 2 hours and the
reaction was stirred under nitrogen at room temperature for 18
hours, concentrated under reduced pressure and the residue was
suspended in methanol (250 ml). After filtration, the filtrates
were concentrated under reduced pressure, the residue was dissolved
in ethyl acetate (400 ml) and washed with water (50 ml). The
organic phase was filtered, the filtrate was dried over MgSO.sub.4
and concentrated under reduced pressure. The filtered solid and
residue from the filtrates were combined to give
5-isobutyl-4-nitro-1H-pyrazol-3-carbo- xylic acid amide (4.8 g) as
an off-white solid. .sup.1H NMR (400 MHz, DMSO-D6):
.delta.=13.61-13.81 (1H, brs), 7.80-7.96 (1H, brs), 7.50-7.66 (1H,
brs), 2.70-2.76 (2H, d), 1.90-2.01 (1H, m), 0.83-0.88 (6H, d) ppm.
LRMS (electrospray): m/z [M+Na].sup.+ 235, [2M+Na].sup.+ 447,
[M-H].sup.+ 211, [2M-H].sup.+ 423. Anal. Found C, 45.12; H, 5.68;
N, 26.31. C.sub.8H.sub.12N.sub.4O.sub.3 requires C, 45.28; H, 5.70;
N, 26.40%.
Preparation 24
4-Amino-5-isobutyl-1H-pyrazol-3-carboxylic Acid Amide
[0278] 191
[0279] 5-Isobutyl-4-nitro-1H-pyrazol-3-carboxylic acid amide (4.7
g, 22 mmol) and 10% palladium on carbon (300 mg) in ethanol (80 ml)
was stirred under hydrogen (60 psi) at room temperature for 4
hours, and held under nitrogen for 64 hours. The reaction mixture
was filtered and the filtrate was concentrated under reduced
pressure of dichloromethane:methanol (100:0 changing to 95:5 then
90:10, by volume) to give
4-amino-5-isobutyl-1H-pyrazol-3-carboxylic acid amide (3.8 g) as an
off-white solid, which was a mixture of rotamers. .sup.1H NMR (400
MHz, DMSO-D6): .delta.=12.20-12.28 (1H, brs), 7.00-7.10 (1.34H,
brs), 6.80-6.85 (0.66H, brs), 4.27-4.40 (2H, brs), 2.27-2.36 (2H,
d), 1.78-1.88 (1H, m), 0.77-0.84 (6H, d) ppm. LRMS (electrospray):
m/z [M+H].sup.+ 183, [M+Na].sup.+ 205. Anal. Found C, 52.27; H,
7.78; N, 30.59. C.sub.8H.sub.14N.sub.4O requires C, 52.73; H, 7.76;
N, 30.75%.
Preparation 25
1-Cyclopentylethanone
[0280] 192
[0281] Concentrated sulfuric acid (22.4 ml, 420 mmol) was slowly
added to a solution of chromium trioxide (26.3 g, 263 mmol)
dissolved in water (50 ml) at room temperature. After 10 min this
solution was added to 1-cyclopentylethanol (20 g, 175 mmol)
dissolved in acetone (450 ml) maintaining the temperature below
35.degree. C. The addition was continued until a bright orange
colour persisted for 10 minutes. The reaction mixture was quenched
with isopropyl alcohol to destroy excess chromic acid and it was
then neutralised to pH 5 with the portionwise addition of sodium
bicarbonate. After filtration the filtrate was concentrated under
reduced pressure (to 50 ml) and extracted with diethylether
(3.times.300 ml). The combined organic extracts were dried over
MgSO.sub.4 and concentrated under reduced pressure to give
1-cyclopentylethanone (16.7 g) as a colourless oil. .sup.1H NMR
(400 MHz, CDCl.sub.3): .delta. 2.80-2.90 (1H, quin), 2.18 (3H, s),
1.53-1.86 (8H, 2xm) ppm.
Preparation 26
3-Cyclopentyl-3-oxo-propionic Acid Ethyl Ester
[0282] 193
[0283] Sodium pellets (3.1 g, 135 mmol) were dissolved in ethanol
(100 ml) under nitrogen at room temperature and a solution of
diethyloxalate (18.4 ml, 135 mmol) and 1-cyclopentylethanone (16.7
g, 149 mmol) was added dropwise at room temperature over 30
minutes. The reaction was diluted with ethanol (100 ml), heated to
60.degree. C. and stirred at this temperature for 2 hours. After
cooling to room temperature the reaction was poured onto ice-cold
2N HCl (200 ml) and extracted with diethylether (300 ml) and ethyl
acetate (300 ml). The combined organic extracts were dried over
MgSO.sub.4, concentrated under reduced pressure and the residue was
purified by flash column chromatography on silica gel eluting with
pentane: ethyl acetate (6:1, by volume) to give
3-cyclopentyl-3-oxo-propionc acid ethyl ester (23.8 g) as an orange
oil. .sup.1H NMR (400 MHz, CDCl.sub.3): .delta.=14.38-14.65 (1H,
brs), 6.83 (1H, s), 4.30-4.39 (2H, quart), 2.82-2.92 (1H, quin),
1.83-1.96 (2H, m), 1.57-1.83 (6H, 2xm), 1.33-1.40 (3H, t) ppm. LRMS
(electrospray): m/z [M-H].sup.+ 211.
Preparation 27
5-Cyclopentyl-1H-pyrazol-3-carboxylic Acid Ethyl Ester
[0284] 194
[0285] Hydrazine hydrate (5.8 ml, 117 mmol) was added to a solution
of 3-cyclopentyl-3-oxo-propionc acid ethyl ester (23.7 g, 112 mmol)
in ethanol (100 ml) at room temperature under nitrogen. The
reaction was allowed to proceed at room temperature for 18 hours,
then heated to 50.degree. C. and held at this temperature for 4
hours. The solvent was removed under reduced pressure and the
residue was partitioned between dichloromethane (300 ml) and water
(300 ml) and the aqueous phase was removed. The organic phase was
washed with water (2.times.200 ml), dried over MgSO.sub.4 and
concentrated under reduced pressure. The residue was purified by
flash column chromatography on silica gel eluting with a solvent
gradient of pentane:ethyl acetate (4:1, by volume) to give
5-cyclopentyl-1H-pyrazol-3-carboxylic acid ethyl ester (17.1 g) as
a pale yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3):
.delta.=10.40-10.60 (1H, brs), 6.58 (1H, s), 4.30-4.38 (2H, quart),
3.01-3.10 (1H, quin), 2.00-2.10 (2H, m), 1.56-1.80 (6H, 2xm),
1.33-1.39 (3H, t) ppm. LRMS (electrospray): m/z [M+H].sup.+ 209,
[M+Na].sup.+ 231. Anal. Found C, 63.40; H, 7.75; N, 13.41.
C.sub.11H.sub.16N.sub.2O.sub.2 requires C, 63.44; H, 7.74; N,
13.45%.
Preparation 28
5-Cyclopentyl-1H-pyrazol-3-carboxylic Acid
[0286] 195
[0287] 5-Cyclopentyl-1H-pyrazol-3-carboxylic acid ethyl ester (17.1
g, 82 mmol) and 1M NaOH solution (205 ml, 205 mmol) were dissolved
in 1,4-dioxan (300 ml) and the reaction was heated to 50.degree. C.
under nitrogen and stirred for 3 hours. The reaction mixture was
cooled, adjusted to pH 2 using concentrated hydrochloric acid and
the solvent was removed under reduced pressure. The residual solid
was azeotrped with toluene (2.times.30 ml), dissolved in ethyl
acetate (500 ml) and washed with water (200 ml). The aqueous phase
was removed, extracted with ethyl acetate (2.times.200 ml) and the
combined organic extracts were dried over MgSO.sub.4. The solvent
was removed under reduced pressure and the residue was azeotrped
with dichloromethane (2.times.50 ml) to give
5-cyclopentyl-1H-pyrazol-3-carboxylic acid (13 g) as a white solid.
.sup.1H NMR (400 MHz, DMSO-D6): .delta.=12.75-12.88 (2H, brs), 6.43
(1H, s), 2.97-3.08 (1H, quin), 1.91-2.02 (2H, m), 1.50-1.76 (6H,
2xm) ppm. LRMS (electrospray): m/z [M-H].sup.+ 179. Anal. Found C,
59.72; H, 6.74; N, 15.37. C.sub.9H.sub.12N.sub.2O.sub.2 requires C,
59.99; H, 6.71; N, 15.55%.
Preparation 29
5-Cyclopentyl-4-nitro-1H-pyrazol-3-carboxylic Acid
[0288] 196
[0289] 5-Cyclopentyl-1H-pyrazol-3-carboxylic acid (13 g, 72.1 mmol)
was added portionwise to concentrated sulfuric acid (75 ml) at room
temperature with stirring. The reaction mixture was then heated to
60.degree. C. and concentrated nitric acid (70%, 12.7 ml, 202 mmol)
was added dropwise, keeping the temperature at 60.degree. C. The
reaction was then stirred at 60.degree. C. for 3 hours, cooled to
room temperature and poured onto 50 ml of ice with stirring. After
15 minutes the precipitate was isolated by filtration, washed with
water and dried under reduced pressure to give
5-cyclopentyl-4-nitro-1H-pyrazol-3-carboxylic acid (7.1 g) as a
yellow solid. .sup.1H NMR (400 MHz, DMSO-D6): .delta.=14.00-14.41
(1H, brs), 13.28-13.85 (1H, brs), 3.20-3.56 (1H, brs, partially
masked by solvent), 1.96-2.10 (2H, m), 1.54-1.80 (6H, 2xm) ppm.
LRMS (electrospray): m/z [M-H].sup.+ 224, [2M-H].sup.+ 449. Anal.
Found C, 43.83; H, 5.35; N, 16.94. C.sub.9H.sub.11N.sub.3O.sub.4.
1.2 mol H.sub.2O requires C, 43.80; H, 5.47; N, 17.02%.
Preparation 30
5-Cyclopentyl-4-nitro-1H-pyrazol-3-carboxylic Acid Amide
[0290] 197
[0291] Oxalyl chloride (7.65 ml, 87.7 mmol) was added dropwise to a
suspension of 5-isopropyl-4-nitro-1H-pyrazol-3-carboxylic acid
(6.58 g, 29.2 mmol) in dichloromethane (100 ml) containing
dimethylformamide (0.5 ml) under nitrogen at 0.degree. C. The
reaction was stirred at 0.degree. C. for 1 hours, allowed to warm
to room temperature and stirred for a further 2 hours. The solvent
was removed under reduced pressure, the residue was azeotroped with
dichloromethane (2.times.50 ml) and dissolved in toluene (100 ml).
Ammonia gas was bubbled into the solution for 2 hours and the
reaction was stirred under nitrogen at room temperature for 18
hours, concentrated under reduced pressure and purified by flash
column chromatography on silica gel eluting with a solvent gradient
of dichloromethane:methanol (95:5 changing to 90:10, by volume) to
give 5-cyclopentyl-4-nitro-1H-pyrazol-3-carboxylic acid amide (5.48
g) as a yellow solid. .sup.1H NMR (400 MHz, DMSO-D6):
.delta.=13.67-13.79 (1H, brs), 7.88-8.03 (1H, brs), 7.59-7.77 (1H,
brs), 3.46-3.60 (1H, quin), 1.97-2.11 (2H, m), 1.58-1.81 (6H, 2xm)
ppm. LRMS (electrospray): m/z [M-H].sup.+ 223, [2M-H].sup.+ 447.
Anal. Found C, 56.12; H, 7.39; N, 27.55.
C.sub.9H.sub.12N.sub.4O.sub.3. 0.2 mol acetone requires C, 56.01;
H, 7.44; N, 27.22%.
Preparation 31
4-Amino-5-cyclopentyl-1H-pyrazol-3-carboxylic Acid Amide
[0292] 198
[0293] 5-cyclopentyl-4-nitro-1H-pyrazol-3-carboxylic acid amide
(4.48 g, 20 mmol) and 10% palladium on carbon (800 mg) in ethanol
(50 ml) were stirred under hydrogen (50 psi) at room temperature
for 18 hours. The reaction mixture was filtered through arbocel and
the solid was washed with ethanol (50 ml), methanol (50 ml),
dichloromethane (50 ml), and ethyl acetate (50 ml). The filtrate
was concentrated under reduced pressure and the residue was
purified by flash column chromatography on silica gel eluting with
dichloromethane: methanol (9:1, by volume) to give
4-amino-5-cyclopentyl-1H-pyrazol-3-carboxylic acid amide (4.0 g) as
an off-white solid which was a mixture of rotamers. .sup.1H NMR
(400 MHz, DMSO-D6): .delta.=12.20-12.31 (0.75H, brs), 11.78-11.87
(0.25H, brs), 7.02-7.18 (1.5H, brs), 6.80-6.93 (0.5H, brs),
4.22-4.56 (2H, 2xbrs), 2.92-3.02 (1H, quin), 1.79-1.96 (2H, m),
1.48-1.78 (6H, 2xm) ppm. LRMS (electrospray): m/z [M-H].sup.+ 193.
Anal. Found C, 56.12; H, 7.39; N, 27.55. C.sub.9H.sub.14N.sub.4O.
0.2 mol acetone requires C, 56.01; H, 7.44; N, 27.22%.
Preparation 32
(3-Benzyloxy-phenyl)-acetic Acid Benzyl Ester
[0294] 199
[0295] 3-Hydroxy-phenyl-acetic acid (15.3 g, 101 mmol), benzyl
bromide (36.2 g, 202 mmol) and potassium carbonate (29.2 g, 202
mmol) were suspended in dimethylformamide (300 ml) and the reaction
was heated to reflux under nitrogen for 44 hours. The reaction
mixture was cooled, filtered and the filtrate was concentrated
under reduced pressure. The residue was partitioned between ethyl
acetate (200 ml) and water (200 ml), and the aqueous phase was
extracted with ethyl acetate (2.times.200 ml). The combined organic
extracts were washed with brine (200 ml), dried over
Na.sub.2SO.sub.4 and the solvent was removed under reduced
pressure. The residue was purified by flash column chromatography
on silica gel eluting with pentane:ethyl acetate (95:5, by volume)
to give (3-benzyloxy-phenyl)-acetic acid benzyl ester (10.7 g) as a
white solid.
Preparation 33
(3-Benzyloxy-phenyl)-acetic Acid
[0296] 200
[0297] 1N Sodium hydroxide solution (35 ml, 35 mmol) was added to a
solution of (3-benzyloxy-phenyl)-acetic acid benzyl ester (5.3 g,
16 mmol) in methanol (350 ml) at room temperature under nitrogen.
The reaction was heated to reflux for 2 hours, and the solvent was
removed under reduced pressure. The residue was dissolved in water
(500 ml) and extracted with ether (3.times.350 ml). The aqueous
phase was acidified to pH 1 with concentrated hydrochloric acid and
the resultant precipitate was isolated by filtration and dried
under vacuum to give (3-benzyloxy-phenyl)-acetic acid (3.08 g) as a
white solid. mp 127-129.degree. C. .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta.=7.26-7.43 (5H, m), 7.20-7.26 (1H, m, partially
masked by solvent), 6.84-6.96 (3H, m+s), 5.04 (2H, s), 3.62 (2H, s)
ppm. LRMS (electrospray): m/z [M-H].sup.+ 241. Anal. Found C,
74.21; H, 5.82. C.sub.15H.sub.14O requires C, 74.36; H, 5.82%.
Preparation 34
(4-hydroxy-3-methoxy-phenyl)-acetic Acid Methyl Ester
[0298] 201
[0299] Concentrated sulfuric acid (12 ml) was added to a solution
of (4-hydroxy-3-methyoxy-phenyl)-acetic acid (22.5 g, 123 mmol) in
methanol (450 ml) at room temperature, and the reaction was heated
to 90.degree. C. for 2.45 hours. The reaction was then cooled to
room temperature and stirred for 18 hours, and the solvent was
removed under reduced pressure. The residue was suspended in ice
water (300 ml) and extracted with diethylether (2.times.300 ml).
The combined organic extracts were washed with saturated sodium
bicarbonate solution (2.times.100 ml), brine (100 ml), dried over
Na.sub.2SO.sub.4 and concentrated under reduced pressure. The
residue was purified by flash column chromatography on silica gel
eluting with a solvent gradient of cyclohexane:ethyl acetate (80:20
changing to 70:30, 60:40 and finally 1:1, by volume) to give
(4-hydroxy-3-methoxy-phenyl)-acetic acid methyl ester (23 g) as a
yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3): .delta.=6.82-6.85
(1H, d), 6.80 (1H, s), 6.76-6.79 (1H, d), 5.49 (1H, s), 3.86 (3H,
s), 3.66 (3H, s), 3.53 (2H, s) ppm. LRMS (electrospray): m/z
[M+Na].sup.+ 219.
Preparation 35
(4-Cyclopentyloxy-3-methoxy-phenyl)-acetic Acid Methyl Ester
[0300] 202
[0301] Cyclopentanol (7.7 ml, 85 mmol) and triphenylphosphine (28
g, 107 mmol) were added to a solution of
(4-hydroxy-3-methyoxy-phenyl)-acetic acid methyl ester (14 g, 71
mmol) in tetrahydrofuran (280 ml) under nitrogen at 0.degree. C.
Diethylazodicarboxylate (15.7 ml, 100 mmol) was then added dropwise
and the reaction was allowed to warm to room temperature and
stirred for 44 hours. The solvent was removed under reduced
pressure, pentane (200 ml) was added and the suspension was
filtered. The filtrate was concentrated under reduced pressure and
purified by flash column chromatography on silica gel eluting with
a solvent gradient of cyclohexane:ethyl acetate (90:10 changing to
85:15, by volume) to give
(4-cyclopentyloxy-3-methoxy-phenyl)-acetic acid methyl ester (12.4
g) as a colourless oil. .sup.1H NMR (400 MHz, CD.sub.3OD):
.delta.=6.79-6.85 (2H, m), 6.73-6.79 (1H, d), 4.73-4.79 (1H, brs),
3.79 (3H, s), 3.64 (3H, s), 3.53 (2H, s), 1.74-1.89 (6H, m),
1.56-1.67 (2H, m) ppm. LRMS (electrospray): m/z [M+Na].sup.+ 287.
Anal. Found C, 68.01; H, 7.74. C.sub.15H.sub.20O.sub.4 requires C,
68.16; H, 7.63%.
Preparation 36
(4-Cyclopentyloxy-3-methoxy-phenyl)-acetic Acid
[0302] 203
[0303] Sodium hydroxide (4.75 g, 119 mmol) was added to a solution
of (4-cyclopentyloxy-3-methoxy-phenyl)-acetic acid methyl ester
(12.4, 46.9 mmol) in methanol)100 ml)/water (100 ml) and the
reaction was stirred at room temperature for 3.5 hours. The
methanol was removed under reduced pressure and the aqueous phase
was washed with diethylether (100 ml) then acidified to pH 2 using
concentrated hydrochloric acid. This was then extracted with ethyl
acetate (2.times.200 ml) and the combined organic extracts were
washed with brine (100 ml), dried over Na.sub.2SO.sub.4 and
concentrated under reduced pressure to give
(4-cyclopentyloxy-3-methoxy-p- henyl)-acetic acid (11.1 g) as a
white solid. .sup.1H NMR (400 MHz, CD.sub.3OD): .delta.=6.87 (1H,
s), 6.81-6.86 (1H, d), 6.76-6.80 (1H, d), 4.75-4.79 (1H, brs), 3.78
(3H, s), 3.49 (2H, s), 1.71-1.89 (6H, m), 1.56-1.64 (2H, m) ppm.
LRMS (electrospray): m/z [M-H].sup.+ 249, [2M-H].sup.+ 499. Anal.
Found C, 67.15; H, 7.25. C.sub.14H.sub.18O.sub.4 requires C, 67.18;
H, 7.25%.
Preparation 37
2,4-Dimethyl-phenyl-acetic Acid
[0304] 204
[0305] 2,4-Dimethylbenzylcyanide (70 g, 0.48 mol) was mixed with
water (134 ml) and concentrated sulfuric acid (106 ml, 1.98 mol)
was added slowly. The reaction was heated to reflux for 3 hours,
then cooled to room temperature over 18 hours. The mixture was
poured onto crushed ice (500 ml), stirred for one hour and the
resulting precipitate was isolated by filtration. After washing
with water the solid was dissolved in 1.2M sodium hydroxide
solution (500 ml), extracted with dichloromethane (2.times.250 ml)
and the aqueous phase was treated with decolourising carbon (2 g)
at reflux for 10 min and filtered hot through hyflo supercel. The
filtrate was then acidified with concentrated hydrochloric acid and
the resulting precipitate was isolated by filtration, washed with
water and dried under vacuum to give 2,4-dimethyl-phenyl-acetic
acid (52.6 g) as a white solid. .sup.1H NMR (250 MHz,
CD.sub.3OD/D.sub.2O): .delta.=6.88-7.03 (3H, m), 3.48-3.68 (2H, s),
2.23 (6H, s) ppm.
Preparation 38
Benzene Sulfonic Acid 2-chloro-ethyl Ester
[0306] 205
[0307] 2-Chloroethanol (1168 g, 975 mol) and benzene sulfonyl
chloride (2780 g, 2015 mol) were stirred together at -5.degree. C.
and pyridine (2158 g, 2200 mol) was added over a 3 hours period,
maintaining the temperature below 0.degree. C. The reaction was
stirred for a further 3 hours at -5.degree. C. to 0.degree. C. and
was then allowed to warm to room temperature over 18 hours. After
pouring into a mixture of ice (10 liters) and water (10 liters) the
reaction was stirred for 15 minutes, extracted with ether (10
liters) and the organic phase was washed with 5N HCl (2.times.2
liters) and water (2.times.4 liters). It was then dried over
MgSO.sub.4 and concentrated under reduced pressure to give benzene
sulfonic acid 2-chloro-ethyl ester (1921 g) as an orange oil.
.sup.1H NMR (250 MHz, CDCl.sub.3): .delta.=7.78-8.02 (2H, m),
7.58-7.78 (3H, m), 4.20-4.45 (2H, t), 3.60--3.81 (2H, t) ppm.
Preparation 39
2-Hydroxy-phenyl-acetic Acid Ethyl Ester
[0308] 206
[0309] 2-Hydroxy-phenyl-acetic acid (30.4 g, 0.2 mol) was dissolved
in chloroform (200 ml) and thionyl chloride (50 ml, 0.2 mol) was
added. The reaction was gently refluxed for 2 hours, upon which the
mixture was concentrated under reduced pressure. The residue was
slowly poured into ethanol (200 ml) maintaining a temperature of
10.degree. C. to 20.degree. C. The solvent was removed under
reduced pressure and the residue was purified by thermal
distillation to give 2-hydroxy-phenyl-acetic acid ethyl ester (31.6
g) as a yellow oil. Bp 146-150.degree. C. .nu..sub.max (thin film)
1710 cm.sup.1 (C.dbd.O, ester).
Preparation 40
[2-(2-Chloro-ethoxy)-phenyl]-acetic Acid Ethyl Ester
[0310] 207
[0311] 50% Sodium hydride in mineral oil (8.11 g, 169 mmol) was
added portionwise to a solution of 2-hydroxy-phenyl-acetic acid
ethyl ester (30.4 g, 169 mmol) in dimethylformamide (100 ml). After
the initial effervescence had ended the reaction was heated to
100.degree. C. for 10 minutes and was cooled to room temperature. A
solution of benzene sulfonic acid 2-chloro-ethyl ester (37.2 g, 169
mmol) in dimethylformamide (5 ml) was then added and the reaction
was heated to 100.degree. C. for one hour, and allowed to cool to
room temperature over 18 hours. The reaction mixture was
partitioned between diethylether (300 ml) and water (300 ml) and
the organic phase was removed and washed with water (100 ml), dried
over MgSO.sub.4 and the solvent was removed under reduced pressure.
The residue was purified by thermal distillation to give
[2-(2-chloro-ethoxy)-phenyl]-acetic acid ethyl ester (22.0 g) as a
pale yellow oil. Bp 170.degree. C. to 180.degree. C. .nu..sub.max
(thin film) 1735 cm.sup.-1 (C.dbd.O, ester); no O--H stretch. Anal.
Found C, 59.35; H, 6.29. C.sub.12H.sub.15ClO.sub.3 requires C,
59.38; H, 6.23%.
Preparation 41
[2-(2-Imidazol-1-yl-ethoxy)-phenyl]-acetic Acid
[0312] 208
[0313] [2-(2-Imidazol-1-yl-ethoxy)-phenyl]-acetic acid ethyl ester
(3.5 g, 113 mmol) was stirred in 50% aqueous hydrochloric acid (20
ml) at 100.degree. C. for 6 hours. After cooling to room
temperature the solvent was removed under reduced pressure and the
residue was recrystallised from isopropylalcohol to give
[2-(2-Imidazol-1-yl-ethoxy)-phenyl]-acetic acid (2.73 g) as a white
solid. Mp 146-147.degree. C. .nu..sub.max (thin film) 3410 (O--H),
1722 cm.sup.-1 (C.dbd.O, acid). Anal. Found C, 54.89; H, 5,25; N,
9.80. C.sub.13H.sub.14N.sub.2O.sub.3. 1 mol HCl requires C, 55.22;
H, 5.35; N, 9.91%.
Preparation 42
5-Cyclopentyl-4-[2-(2-trifluoromethoxy-phenyl)-acetylamino]-1H-pyrazole-3--
carboxylic Acid Amide
[0314] 209
[0315] Carbonyldiimidazole (84 mg, 0.515 mmol) was added to a
solution of 2-triflouoromethyoxy-phenyl-acetic acid (113 mg, 0.515
mmol) in tetrahydrofuran (4 ml) under nitrogen at room temperature,
and the mixture was stirred for 3 hours.
4-Amino-5-cyclopentyl-1H-pyrazol-3-carbo- xylic acid amide (100 mg,
0.515 mmol) was then added and the reaction was stirred for 18
hours. The reaction mixture was diluted with water (20 ml),
acidified to pH 2 with 2N HCl and extracted with ethyl acetate
(2.times.20 ml). The combined organic extracts were dried over
MgSO.sub.4 and concentrated under reduced pressure to give
5-cyclopentyl-4-[2-(2-tri-
fluoromethoxy-phenyl)-acetylamino]-1H-pyrazole-3-carboxylic acid
amide (120 mg) as an off-white solid. LRMS (electrospray): m/z
[M+H].sup.+ 397, [M-H].sup.+ 395.
Preparation 43
5-Isobutyl-4-[2-(2-trifluoromethoxy-phenyl)-acetylamino]-1H-pyrazole-3-car-
boxylic Acid Amide
[0316] 210
[0317] Carbonyldiimidazole (84 mg, 0.515 mmol) was added to a
solution of 2-triflouoromethyoxy-phenyl-acetic acid (113 mg, 0.515
mmol) in tetrahydrofuran (4 ml) under nitrogen at room temperature,
and the mixture was stirred for 3 hours.
4-Amino-5-isobutyl-1H-pyrazol-3-carboxyl- ic acid amide (100 mg,
0.515 mmol) was then added and the reaction was stirred for 18
hours. The reaction mixture was diluted with water (20 ml),
acidified to pH2 with 2N HCl and extracted with ethyl acetate
(2.times.20 ml). The combined organic extracts were dried over
MgSO.sub.4 and concentrated under reduced pressure to give
5-isobutyl-4-[2-(2-triflu-
oromethoxy-phenyl)-acetylamino]-1H-pyrazole-3-carboxylic acid amide
(142 mg) as an off-white solid. LRMS (electrospray): m/z
[M+H].sup.+ 385, [M-H]+383.
Preparation 44
5-Pyridine-3-yl-4-[2-(2-trifluoromethoxy-phenyl)-acetylamino]-1H-pyrazole--
3-carboxylic Acid Amide
[0318] 211
[0319] Carbonyldiimidazole (144 mg, 0.886 mmol) was added to a
solution of 2-triflouoromethyoxy-phenyl-acetic acid (195 mg, 0.886
mmol) in tetrahydrofuran (5 ml) under nitrogen at room temperature,
and the mixture was stirred for one hour.
4-Amino-5-cyclopropyl-1H-pyrazol-3-carb- oxylic acid amide (180 mg,
0.886 mmol) was then added and the reaction was stirred for 18
hours. The reaction mixture was diluted with brine (20 ml) and
extracted with ethyl acetate (2.times.20 ml). The combined organic
extracts were dried over MgSO.sub.4 and concentrated under reduced
pressure to give
5-pyridine-3-yl-4-[2-(2-trifluoromethoxy-phenyl)-acetyla-
mino]-1H-pyrazole-3-carboxylic acid amide (345 mg) as an off-white
solid. LRMS (electrospray): m/z [M+Na].sup.+ 428, [M-H].sup.+
404.
* * * * *