U.S. patent application number 10/730403 was filed with the patent office on 2004-10-21 for aromatic sulfone hydroxamic acids and their use as protease inhibitors.
Invention is credited to Barta, Thomas E., Becker, Daniel P., Bedell, Louis J., Boehm, Terri L., Carroll, Jeffery N., DeCrescenzo, Gary A., Fobian, Yvette M., Freskos, John N., Getman, Daniel P., Hockerman, Susan L., Howard, Carol P., Kassab, Darren J., Kolodziej, Stephen A., Li, Madeleine H., McDonald, Joseph J., Mischke, Deborah A., Rico, Joseph G., Stehle, Nathan W., Tollefson, Michael B., Vernier, William F., Villamil, Clara I..
Application Number | 20040209914 10/730403 |
Document ID | / |
Family ID | 25535599 |
Filed Date | 2004-10-21 |
United States Patent
Application |
20040209914 |
Kind Code |
A1 |
Barta, Thomas E. ; et
al. |
October 21, 2004 |
Aromatic sulfone hydroxamic acids and their use as protease
inhibitors
Abstract
This invention is directed to aromatic sulfone hydroxamates
(also known as aromatic sulfone hydroxamic acids) and salts thereof
that, inter alia, tend to inhibit matrix metalloproteinase (also
known as matrix metalloprotease or MMP) activity and/or aggrecanase
activity. This invention also is directed to a treatment method
that comprises administering such a compound or salt in an
MMP-inhibiting and/or aggrecanase-inhibiting effective amount to an
animal, particularly a mammal having (or disposed to having) a
pathological condition associated with MMP activity and/or
aggrecanase activity.
Inventors: |
Barta, Thomas E.; (Wilmette,
IL) ; Becker, Daniel P.; (Glenview, IL) ;
Bedell, Louis J.; (Prospect Heights, IL) ; Boehm,
Terri L.; (Ballwin, MO) ; Carroll, Jeffery N.;
(St. Louis, MO) ; DeCrescenzo, Gary A.; (St.
Charles, MO) ; Fobian, Yvette M.; (Wildwood, MO)
; Freskos, John N.; (Clayton, MO) ; Getman, Daniel
P.; (Chesterfield, MO) ; Hockerman, Susan L.;
(Kirkwood, MO) ; Howard, Carol P.; (Fenton,
MO) ; Kassab, Darren J.; (O'Fallon, MO) ;
Kolodziej, Stephen A.; (Ballwin, MO) ; Li, Madeleine
H.; (Vernon Hills, IL) ; McDonald, Joseph J.;
(Wildwood, MO) ; Mischke, Deborah A.; (Defiance,
MO) ; Rico, Joseph G.; (O'Fallon, MO) ;
Stehle, Nathan W.; (Grafton, WI) ; Tollefson, Michael
B.; (Dardenne Prairie, MO) ; Vernier, William F.;
(St. Louis, MO) ; Villamil, Clara I.; (Glenview,
IL) |
Correspondence
Address: |
HARNESS, DICKEY, & PIERCE, P.L.C
7700 BONHOMME, STE 400
ST. LOUIS
MO
63105
US
|
Family ID: |
25535599 |
Appl. No.: |
10/730403 |
Filed: |
December 8, 2003 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10730403 |
Dec 8, 2003 |
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09989943 |
Nov 21, 2001 |
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6683093 |
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09989943 |
Nov 21, 2001 |
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09570731 |
May 12, 2000 |
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6750228 |
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Current U.S.
Class: |
514/316 ;
514/326; 546/188; 546/207; 546/229 |
Current CPC
Class: |
A61P 27/16 20180101;
A61P 19/02 20180101; C07D 335/02 20130101; A61P 17/02 20180101;
A61P 25/00 20180101; A61P 41/00 20180101; A61P 9/04 20180101; C07D
233/56 20130101; C07D 409/12 20130101; C07D 231/12 20130101; A61P
1/14 20180101; A61P 35/00 20180101; A61P 29/00 20180101; A61P 43/00
20180101; C07D 405/12 20130101; A61P 9/10 20180101; C07D 401/12
20130101; A61P 11/00 20180101; A61P 1/02 20180101; A61P 27/02
20180101; C07D 401/14 20130101; A61P 25/28 20180101; A61P 7/04
20180101; A61P 19/08 20180101; C07D 249/08 20130101; C07D 413/14
20130101; C07D 309/08 20130101; A61P 17/06 20180101; A61P 3/00
20180101; A61P 7/00 20180101; C07D 417/14 20130101; A61P 1/16
20180101; A61P 1/04 20180101; A61P 7/02 20180101; A61P 9/14
20180101; A61P 37/02 20180101; C07D 211/66 20130101; C07D 401/06
20130101; A61P 17/00 20180101; C07D 405/14 20130101; A61P 31/04
20180101; C07D 409/14 20130101 |
Class at
Publication: |
514/316 ;
514/326; 546/188; 546/207; 546/229 |
International
Class: |
A61K 031/4545; A61K
031/453; C07D 41/02; C07D 45/02 |
Claims
1. A compound or a salt thereof, wherein: the compound corresponds
in structure to the Formula X: 599E is selected from the group
consisting of a bond, --C(O)--, and --S--; Y is selected from the
group consisting of hydrogen, alkyl, alkoxy, haloalkyl, aryl,
arylalkyl, cycloalkyl, heteroaryl, hydroxy, aryloxy, arylalkoxy,
heteroaryloxy, heteroarylalkyl, perfluoroalkoxy,
perfluoroalkylthio, trifluoromethylalkyl, alkenyl, heterocyclyl,
cycloalkyl, trifluoromethyl, alkoxycarbonyl, and aminoalkyl,
wherein: the aryl, heteroaryl, arylalkyl, or heterocyclyl
optionally is substituted with up to 2 substituents independently
selected from the group consisting of alkylcarbonyl, halo, nitro,
arylalkyl, aryl, alkoxy, trifluoroalkyl, trifluoroalkoxy, and
amino, wherein: the amino nitrogen optionally is substituted with
up to 2 substituents independently selected from the group
consisting of alkyl and arylalkyl; and R is selected from the group
consisting of hydrogen, cyano, perfluoroalkyl, trifluoromethoxy,
trifluoromethylthio, haloalkyl, trifluoromethylalkyl,
arylalkoxycarbonyl, aryloxycarbonyl, hydroxy, halo, alkyl, alkoxy,
nitro, thiol, hydroxycarbonyl, aryloxy, arylthio, arylalkyl, aryl,
arylcarbonylamino, heteroaryloxy, heteroarylthio, heteroarylalkyl,
cycloalkyl, heterocylyloxy, heterocylylthio, heterocylylamino,
cycloalkyloxy, cycloalkylthio, heteroarylalkoxy,
heteroarylalkylthio, arylalkoxy, arylalkylthio, arylalkylamino,
heterocylyl, heteroaryl, arylazo, hydroxycarbonylalkoxy,
alkoxycarbonylalkoxy, alkylcarbonyl, arylcarbonyl,
arylalkylcarbonyl, alkylcarbonyloxy, arylalkylcarbonyloxy,
hydroxyalkyl, hydroxyalkoxy, alkylthio, alkoxyalkylthio,
alkoxycarbonyl, aryloxyalkoxyaryl, arylthioalkylthioaryl,
aryloxyalkylthioaryl, arylthioalkoxyaryl, hydroxycarbonylalkoxy,
hydroxycarbonylalkylthio, alkoxycarbonylalkoxy,
alkoxycarbonylalkylthio, amino, aminocarbonyl, and aminoalkyl,
wherein: the amino nitrogen optionally is substituted with: up two
substituents that are independently selected from the group
consisting of alkyl, aryl, heteroaryl, arylalkyl, cycloalkyl,
arylalkoxycarbonyl, alkoxycarbonyl, arylcarbonyl,
arylalkylcarbonyl, heteroarylcarbonyl, heteroarylalkylcarbonyl, and
alkylcarbonyl, or two substituents such that the two substituents,
together with the amino nitrogen, form a 5- to 8-member
heterocyclyl or heteroaryl ring that: contains from zero to two
additional heteroatoms that are independently selected from the
group consisting of nitrogen, oxygen, and sulfur, optionally is
substituted with up to two substituents independently selected from
the group consisting of aryl, alkyl, heteroaryl, arylalkyl,
heteroarylalkyl, hydroxy, alkoxy, alkylcarbonyl, cycloalkyl,
heterocylylalkyl, alkoxycarbonyl, hydroxyalkyl, trifluoromethyl,
benzofused heterocylylalkyl, hydroxyalkoxyalkyl,
arylalkoxycarbonyl, hydroxycarbonyl, aryloxycarbonyl, benzofused
heterocylylalkoxy, benzofused cycloalkylcarbonyl,
heterocyclylalkylcarbonyl, and cycloalkylcarbonyl, the
aminocarbonyl nitrogen is: unsubstituted, the reacted amine of an
amino acid, substituted with one or two substituents independently
selected from the group consisting of alkyl, hydroxyalkyl,
hydroxyheteroarylalkyl, cycloalkyl, arylalkyl,
trifluoromethylalkyl, heterocylylalkyl, benzofused
heterocylylalkyl, benzofused cycloalkyl, and N,N-dialkylsubstituted
alkylamino-alkyl, or substituted with two substituents such that
the two substituents, together with the aminocarbonyl nitrogen,
form a 5- to 8-member heterocyclyl or heteroaryl ring that
optionally is substituted with up to two substituents independently
selected from the group consisting of alkyl, alkoxycarbonyl, nitro,
heterocylylalkyl, hydroxy, hydroxycarbonyl, aryl, arylalkyl,
heteroaralkyl, and amino, wherein the amino nitrogen optionally is
substituted with: two substituents independently selected from the
group consisting of alkyl, aryl, and heteroaryl; or two
substituents such that the two substituents, together with the
amino nitrogen, form a 5- to 8-member heterocyclyl or heteroaryl
ring, and the aminoalkyl nitrogen optionally is substituted with:
up to two substituents independently selected from the group
consisting of alkyl, aryl, arylalkyl, cycloalkyl,
arylalkoxycarbonyl, alkoxycarbonyl, and alkylcarbonyl, or two
substituents such that the two substituents, together with the
aminoalkyl nitrogen, form a 5- to 8-member heterocyclyl or
heteroaryl ring.
2. A compound or salt according to claim 1, wherein R is halo.
3. A compound or salt according to claim 1, wherein the compound
corresponds in structure to Formula XA: 600
4. A compound or salt according to claim 3, wherein the salt is a
pharmaceutically acceptable salt.
5. A compound or salt according to claim 3, wherein Y is selected
from the group consisting of aryl, arylalkyl, cycloalkyl,
heteroaryl, aryloxy, arylalkoxy, heteroaryloxy, heteroarylalkyl,
heterocyclyl, and cycloalkyl, wherein: the aryl, heteroaryl,
arylalkyl, or heterocyclyl optionally is substituted with up to 2
substituents independently selected from the group consisting of
alkylcarbonyl, halo, nitro, arylalkyl, aryl, alkoxy,
trifluoroalkyl, trifluoroalkoxy, and amino, wherein: the amino
nitrogen optionally is substituted with up to 2 substituents
independently selected from the group consisting of alkyl and
arylalkyl.
6. A compound or salt according to claim 3, wherein E is a
bond.
7. A compound or salt according to claim 3, wherein E is
--C(O)--.
8. A compound or salt according to claim 3, wherein E is --S--.
9-13 (canceled)
14. A compound or a salt thereof, wherein: the compound corresponds
in structure to Formula X: 601E is selected from the group
consisting of a bond, --C(O)--, and --S--; and Y is selected from
the group consisting of cycloalkyl, 2,3-dihydroindolyl,
heterocyclyl, aryl, heteroaryl, arylalkyl, and heteroarylalkyl,
wherein: any such substituent optionally is substituted with one or
more optionally substituted substituents independently selected
from the group consisting of halogen, hydroxy, keto, alkyl,
haloalkyl, hydroxyalkyl, alkenyl, alkoxy, alkylcarbonyl,
haloalkoxy, alkylthio, alkoxyalkyl, alkoxycarbonylalkyl,
cycloalkyl, cycloalkylalkyl, cycloalkyloxy, cycloalkylalkoxy,
cycloalkylalkoxyalkyl, aryl, arylalkyl, arylalkoxy, heterocyclyl,
heterocyclylalkyl, heteroaryl, heteroarylcarbonyl,
heterocyclylcarbonylalkyl, alkylsulfonyl, amino, aminoalkyl, and
aminocarbonyl, wherein: any such substituent optionally is
substituted with one or more substituents independently selected
from the group consisting of halogen, nitro, alkyl, haloalkyl,
alkoxy, haloalkoxy, and alkylcarbonyl, and the nitrogen of the
amino, aminoalkyl, or aminocarbonyl optionally is substituted with
up to two substituents independently selected from the group
consisting of alkyl and cycloalkylalkyl; and R is selected from the
group consisting of hydrogen and halogen.
15. A compound or salt according to claim 14, wherein the compound
corresponds in structure to Formula XA: 602
16-25 (canceled).
26. A compound or salt according to claim 15, wherein E is
--C(O)--.
27-28 (canceled).
29. A compound or salt according to claim 26, wherein: Y is
selected from the group consisting of heterocyclyl, aryl,
heteroaryl, and arylmethyl, wherein: any such substituent
optionally is substituted with one or more substituents
independently selected from the group consisting of halogen,
hydroxy, C.sub.1-C.sub.6-alkyl, halo-C.sub.1-C.sub.6-alkyl,
hydroxy-C.sub.1-C.sub.6-alkyl, C.sub.2-C.sub.6-alkenyl,
C.sub.1-C.sub.6-alkoxy, C.sub.1-C.sub.6-alkylcarbonyl,
halo-C.sub.1-C.sub.6-alkoxy, C.sub.1-C.sub.6-alkylthio,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxycarbo- nyl-C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyl, aryl, aryl-C.sub.1-C.sub.6-alkyl,
aryl-C.sub.1-C.sub.6-alkoxy, heterocyclyl,
heterocyclyl-C.sub.1-C.sub.6-alkyl, heteroaryl, heteroarylcarbonyl,
heterocyclylcarbonyl-C.sub.1-C.sub.6-alkyl, amino, and
amino-C.sub.1-C.sub.6-alkyl, wherein: any such substituent
optionally is substituted with one or more substituents
independently selected from the group consisting of halogen, nitro,
C.sub.1-C.sub.6-alkyl, halo-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxy, and C.sub.1-C.sub.6-alkylcarbonyl, and the
nitrogen of the amino or amino-C.sub.1-C.sub.6-alkyl optionally is
substituted with up to two substituents independently selected from
the group consisting of C.sub.1-C.sub.6-alkyl and
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alky- l.
30. A compound or salt according to claim 29, wherein Y is phenyl
optionally substituted with one or more substituents independently
selected from the group consisting of halogen, hydroxy,
C.sub.1-C.sub.6-alkyl, halo-C.sub.1-C.sub.6-alkyl,
hydroxy-C.sub.1-C.sub.6-alkyl, C.sub.2-C.sub.6-alkenyl,
C.sub.1-C.sub.6-alkoxy, C.sub.1-C.sub.6-alkylcarbonyl,
halo-C.sub.1-C.sub.6-alkoxy, C.sub.1-C.sub.6-alkylthio,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxycarbo- nyl-C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyl, aryl, aryl-C.sub.1-C.sub.6-alkyl,
aryl-C.sub.1-C.sub.6-alkoxy, heterocyclyl,
heterocyclyl-C.sub.1-C.sub.6-alkyl, heteroaryl, heteroarylcarbonyl,
heterocyclylcarbonyl-C.sub.1-C.sub.6-alkyl, amino, and
amino-C.sub.1-C.sub.6-alkyl, wherein: any such substituent
optionally is substituted with one or more substituents
independently selected from the group consisting of halogen, nitro,
C.sub.1-C.sub.6-alkyl, halo-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxy, and C.sub.1-C.sub.6-alkylcarbonyl, and the
nitrogen of the amino or amino-C.sub.1-C.sub.6-alkyl optionally is
substituted with up to two substituents independently selected from
the group consisting of C.sub.1-C.sub.6-alkyl and
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alky- l.
31. A compound or salt according to claim 29, wherein Y is thienyl
optionally substituted with one or more substituents independently
selected from the group consisting of halogen, hydroxy,
C.sub.1-C.sub.6-alkyl, halo-C.sub.1-C.sub.6-alkyl,
hydroxy-C.sub.1-C.sub.6-alkyl, C.sub.2-C.sub.6-alkenyl,
C.sub.1-C.sub.6-alkoxy, C.sub.1-C.sub.6-alkylcarbonyl,
halo-C.sub.1-C.sub.6-alkoxy, C.sub.1-C.sub.6-alkylthio,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxycarbo- nyl-C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyl, aryl, aryl-C.sub.1-C.sub.6-alkyl,
aryl-C.sub.1-C.sub.6-alkoxy, heterocyclyl,
heterocyclyl-C.sub.1-C.sub.6-alkyl, heteroaryl, heteroarylcarbonyl,
heterocyclylcarbonyl-C.sub.1-C.sub.6-alkyl, amino, and
amino-C.sub.1-C.sub.6-alkyl, wherein: any such substituent
optionally is substituted with one or more substituents
independently selected from the group consisting of halogen, nitro,
C.sub.1-C.sub.6-alkyl, halo-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxy, and C.sub.1-C.sub.6-alkylcarbonyl, and the
nitrogen of the amino or amino-C.sub.1-C.sub.6-alkyl optionally is
substituted with up to two substituents independently selected from
the group consisting of C.sub.1-C.sub.6-alkyl and
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alky- l.
32-33 (canceled).
34. A compound or salt according to claim 26, wherein: Y is
selected from the group consisting of aryl, heteroaryl, arylmethyl,
and heteroarylmethyl, wherein: any such substituent optionally is
substituted with one or more substituents independently selected
from the group consisting of halogen, C.sub.1-C.sub.6-alkyl,
hydroxy-C.sub.1-C.sub.6-alk- yl, C.sub.2-C.sub.6-alkenyl,
C.sub.1-C.sub.6-alkoxy,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyl-- C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyloxy,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alkoxy,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
heterocyclyl-C.sub.1-C.sub.6-alkyl, amino, and
amino-C.sub.1-C.sub.6-alky- l, wherein: the nitrogen of the amino
or amino-C.sub.1-C.sub.6-alkyl optionally is substituted with up to
two substituents independently selected from the group consisting
of C.sub.1-C.sub.6-alkyl.
35. A compound or salt according to claim 34, wherein Y is phenyl
or phenylmethyl, wherein: the phenyl or phenylmethyl optionally is
substituted with one or more substituents independently selected
from the group consisting of halogen, C.sub.1-C.sub.6-alkyl,
hydroxy-C.sub.1-C.sub.6-alkyl, C.sub.2-C.sub.6-alkenyl,
C.sub.1-C.sub.6-alkoxy,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloal- kyloxy,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alkoxy,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
heterocyclyl-C.sub.1-C.sub.6-alkyl, amino, and
amino-C.sub.1-C.sub.6-alky- l, wherein: the nitrogen of the amino
or amino-C.sub.1-C.sub.6-alkyl optionally is substituted with up to
two substituents independently selected from the group consisting
of C.sub.1-C.sub.6-alkyl.
36-40 (canceled).
41. A compound or salt according to claim 34, wherein Y is thienyl
or thienylmethyl, wherein: the thienyl or thienylmethyl optionally
is substituted with one or more substituents independently selected
from the group consisting of halogen, C.sub.1-C.sub.6-alkyl,
hydroxy-C.sub.1-C.sub.6-alkyl, C.sub.2-C.sub.6-alkenyl,
C.sub.1-C.sub.6-alkoxy,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloal- kyloxy,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alkoxy,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
heterocyclyl-C.sub.1-C.sub.6-alkyl, amino, and
amino-C.sub.1-C.sub.6-alky- l, wherein: the nitrogen of the amino
or amino-C.sub.1-C.sub.6-alkyl optionally is substituted with up to
two substituents independently selected from the group consisting
of C.sub.1-C.sub.6-alkyl.
42-46 (canceled).
47. A compound or salt according to claim 15, wherein E is a
bond.
48-49 (canceled).
50. A compound or salt according to claim 47, wherein: Y is
selected from the group consisting of aryl, 2,3-dihydroindolyl,
heterocyclyl, and heteroaryl, wherein: any such substituent
optionally is substituted with one or more substituents
independently selected from the group consisting of halogen, keto,
hydroxy, C.sub.1-C.sub.6-alkyl, C.sub.1-C.sub.6-alkoxy,
halo-C.sub.1-C.sub.6-alkyl, halo-C.sub.1-C.sub.6-alkoxy, aryl,
aminocarbonyl, and C.sub.1-C.sub.6-alkylsulfonyl, wherein: any such
substituent optionally is substituted with one or more substituents
independently selected from the group consisting of halogen,
halo-C.sub.1-C.sub.6-alkyl, and halo-C.sub.1-C.sub.6-alkoxy, and
the nitrogen of the aminocarbonyl optionally is substituted with up
to 2 substituents independently selected from the group consisting
of C.sub.1-C.sub.6-alkyl.
51. A compound or salt according to claim 50, wherein Y is phenyl
optionally substituted with one or more substituents independently
selected from the group consisting of halogen, keto, hydroxy,
C.sub.1-C.sub.6-alkyl, C.sub.1-C.sub.6-alkoxy,
halo-C.sub.1-C.sub.6-alkyl- , halo-C.sub.1-C.sub.6-alkoxy, aryl,
aminocarbonyl, and C.sub.1-C.sub.6-alkylsulfonyl, wherein: any such
substituent optionally is substituted with one or more substituents
independently selected from the group consisting of halogen,
halo-C.sub.1-C.sub.6-alkyl, and halo-C.sub.1-C.sub.6-alkoxy, and
the nitrogen of the aminocarbonyl optionally is substituted with up
to 2 substituents independently selected from the group consisting
of C.sub.1-C.sub.6-alkyl.
52. A compound or salt according to claim 47, wherein: Y is
selected from the group consisting of heteroaryl, aryl, and
heterocyclyl, wherein: any such substituent optionally is
substituted with one or more substituents independently selected
from the group consisting of halogen, C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxy, and aryl, wherein: the aryl optionally is
substituted with one or more substituents independently selected
from the group consisting of halo-C.sub.1-C.sub.6-alkyl.
53. A compound or salt according to claim 50, wherein Y is phenyl
optionally substituted with one or more substituents independently
selected from the group consisting of halogen,
C.sub.1-C.sub.6-alkyl, C.sub.1-C.sub.6-alkoxy, and aryl, wherein:
the aryl optionally is substituted with one or more substituents
independently selected from the group consisting of
halo-C.sub.1-C.sub.6-alkyl.
54. A compound or salt according to claim 15, wherein E is
--S--.
55-56 (canceled).
57. A compound or salt according to claim 54, wherein: Y is
selected from the group consisting of cycloalkyl, aryl, arylmethyl,
and heteroaryl, wherein: any such substituent optionally is
substituted with one or more substituents independently selected
from the group consisting of halogen, halo-C.sub.1-C.sub.6-alkyl,
and halo-C.sub.1-C.sub.6-alkoxy.
58. A compound or salt according to claim 54, wherein: Y is
heteroaryl.
59. A method for treating a pathological condition in an animal,
wherein: the method comprises administering a compound recited in
claim 1 (or a pharmaceutically acceptable salt thereof) to the
animal in an amount effective to treat the condition; the condition
is treatable by inhibiting matrix metalloprotease activity; and the
condition is selected from the group consisting of tissue
destruction, a fibrotic disease, matrix weakening, defective injury
repair, a cardiovascular disease, a pulmonary disease, a kidney
disease, and a central nervous system disease.
60. A method according to claim 59, wherein the compound
corresponds in structure to Formula XA: 603
61. A method according to claim 59, wherein the condition is
selected from the group consisting of osteoarthritis, rheumatoid
arthritis, septic arthritis, tumor invasion, tumor metastasis,
tumor angiogenesis, a decubitis ulcer, a gastric ulcer, a corneal
ulcer, periodontal disease, liver cirrhosis, fibrotic lung disease,
otosclerosis, atherosclerosis, multiple sclerosis, dilated
cardiomyopathy, epidermolysis bullosa, aortic aneurysm, weak injury
repair, an adhesion, scarring, congestive heart failure, coronary
thrombosis, emphysema, proteinuria, and Alzheimer's disease.
62. A method according to claim 59, wherein the condition is
selected from the group consisting of rheumatoid arthritis,
osteoarthritis, septic arthritis, corneal ulceration, epidermal
ulceration, gastric ulceration, tumor metastasis, tumor invasion,
tumor angiogenesis, periodontal disease, proteinuria, Alzheimer's
disease, coronary thrombosis, bone disease, and defective injury
repair.
63 (canceled).
64. A method for treating a pathological condition in an animal,
wherein: the condition is treatable by inhibiting matrix
metalloprotease-2, matrix metalloprotease-9, and/or matrix
metalloprotease-13 activity; and the method comprising
administering a compound recited in claim 1 (or a
pharmaceutically-acceptable salt thereof) to the animal in an
amount effective to inhibit matrix metalloprotease-2, matrix
metalloprotease-9, and/or matrix metalloprotease-13.
65. A method according to claim 64, wherein the compound
corresponds in structure to Formula XA: 604
66. A method according to claim 64, wherein the compound inhibits
matrix metalloprotease-13 selectively over both matrix
metalloprotease-1 and matrix metalloprotease-14.
67-68 (canceled).
69. A method for treating a pathological condition in an animal,
wherein: the method comprises administering a compound recited in
claim 1 (or a pharmaceutically-acceptable salt thereof) to the
animal in an amount effective to or treat the condition, and the
condition is treatable by inhibiting with TNF-.alpha. convertase
activity.
70-71 (canceled).
72. A method for treating a pathological condition in an animal,
wherein: the condition is treatable by inhibiting aggrecanase
activity; and the method comprises administering a compound of
claim 1 (or a pharmaceutically-acceptable salt thereof) to the
animal in an amount effective to or treat the condition.
73. A method according to claim 72, wherein the compound
corresponds in structure to Formula XA: 605
74-78 (canceled).
79. A method for treating a pathological condition in an animal,
wherein: the method comprises administering a compound recited in
claim 14 (or a pharmaceutically acceptable salt thereof) to the
animal in an amount effective to treat the condition; the condition
is treatable by inhibiting matrix metalloprotease activity; and the
condition is selected from the group consisting of tissue
destruction, a fibrotic disease, matrix weakening, defective injury
repair, a cardiovascular disease, a pulmonary disease, a kidney
disease, and a central nervous system disease.
80. A method according to claim 79, wherein the compound
corresponds in structure to Formula XA: 606
81. A method according to claim 79, wherein Y is selected from the
group consisting of heterocyclyl, aryl, heteroaryl, and arylmethyl,
wherein: any such substituent optionally is substituted with one or
more substituents independently selected from the group consisting
of halogen, hydroxy, C.sub.1-C.sub.6-alkyl,
halo-C.sub.1-C.sub.6-alkyl, hydroxy-C.sub.1-C.sub.6-alkyl,
C.sub.2-C.sub.6-alkenyl, C.sub.1-C.sub.6-alkoxy,
C.sub.1-C.sub.6-alkylcarbonyl, halo-C.sub.1-C.sub.6-alkoxy,
C.sub.1-C.sub.6-alkylthio,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxycarbo- nyl-C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyl, aryl, aryl-C.sub.1-C.sub.6-alkyl,
aryl-C.sub.1-C.sub.6-alkoxy, heterocyclyl,
heterocyclyl-C.sub.1-C.sub.6-alkyl, heteroaryl, heteroarylcarbonyl,
heterocyclylcarbonyl-C.sub.1-C.sub.6-alkyl, amino, and
amino-C.sub.1-C.sub.6-alkyl, wherein: any such substituent
optionally is substituted with one or more substituents
independently selected from the group consisting of halogen, nitro,
C.sub.1-C.sub.6-alkyl, halo-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxy, and C.sub.1-C.sub.6-alkylcarbonyl, and the
nitrogen of the amino or amino-C.sub.1-C.sub.6-alkyl optionally is
substituted with up to two substituents independently selected from
the group consisting of C.sub.1-C.sub.6-alkyl and
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alky- l.
82. A method according to claim 79, wherein Y is selected from the
group consisting of aryl, heteroaryl, arylmethyl, and
heteroarylmethyl, wherein: any such substituent optionally is
substituted with one or more substituents independently selected
from the group consisting of halogen, C.sub.1-C.sub.6-alkyl,
hydroxy-C.sub.1-C.sub.6-alkyl, C.sub.2-C.sub.6-alkenyl,
C.sub.1-C.sub.6-alkoxy, C.sub.1-C.sub.6-alkoxy-C-
.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyloxy,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6- -alkoxy,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-
-alkyl, heterocyclyl-C.sub.1-C.sub.6-alkyl, amino, and
amino-C.sub.1-C.sub.6-alkyl, wherein: the nitrogen of the amino or
amino-C.sub.1-C.sub.6-alkyl optionally is substituted with up to
two substituents independently selected from the group consisting
of C.sub.1-C.sub.6-alkyl.
83. A method according to claim 79, wherein the condition is
selected from the group consisting of osteoarthritis, rheumatoid
arthritis, septic arthritis, tumor invasion, tumor metastasis,
tumor angiogenesis, a decubitis ulcer, a gastric ulcer, a corneal
ulcer, periodontal disease, liver cirrhosis, fibrotic lung disease,
otosclerosis, atherosclerosis, multiple sclerosis, dilated
cardiomyopathy, epidermolysis bullosa, aortic aneurysm, weak injury
repair, an adhesion, scarring, congestive heart failure, coronary
thrombosis, emphysema, proteinuria, and Alzheimer's disease.
84. A method for treating a pathological condition in an animal,
wherein: the condition is treatable by inhibiting matrix
metalloprotease-2, matrix metalloprotease-9, and/or matrix
metalloprotease-13 activity; and the method comprising
administering a compound recited in claim 14 (or a
pharmaceutically-acceptable salt thereof) to the animal in an
amount effective to inhibit matrix metalloprotease-2, matrix
metalloprotease-9, and/or matrix metalloprotease-13.
85. A method according to claim 84, herein the compound corresponds
in structure to Formula XA: 607
86. A method according to claim 84, wherein Y is selected from the
group consisting of heterocyclyl, aryl, heteroaryl, and arylmethyl,
wherein: any such substituent optionally is substituted with one or
more substituents independently selected from the group consisting
of halogen, hydroxy, C.sub.1-C.sub.6-alkyl,
halo-C.sub.1-C.sub.6-alkyl, hydroxy-C.sub.1-C.sub.6-alkyl,
C.sub.2-C.sub.6-alkenyl, C.sub.1-C.sub.6-alkoxy,
C.sub.1-C.sub.6-alkylcarbonyl, halo-C.sub.1-C.sub.6-alkoxy,
C.sub.1-C.sub.6-alkylthio,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxycarbo- nyl-C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyl, aryl, aryl-C.sub.1-C.sub.6-alkyl,
aryl-C.sub.1-C.sub.6-alkoxy, heterocyclyl,
heterocyclyl-C.sub.1-C.sub.6-alkyl, heteroaryl, heteroarylcarbonyl,
heterocyclylcarbonyl-C.sub.1-C.sub.6-alkyl, amino, and
amino-C.sub.1-C.sub.6-alkyl, wherein: any such substituent
optionally is substituted with one or more substituents
independently selected from the group consisting of halogen, nitro,
C.sub.1-C.sub.6-alkyl, halo-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxy, and C.sub.1-C.sub.6-alkylcarbonyl, and the
nitrogen of the amino or amino-C.sub.1-C.sub.6-alkyl optionally is
substituted with up to two substituents independently selected from
the group consisting of C.sub.1-C.sub.6-alkyl and
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alky- l.
87. A method according to claim 84, wherein Y is selected from the
group consisting of aryl, heteroaryl, arylmethyl, and
heteroarylmethyl, wherein: any such substituent optionally is
substituted with one or more substituents independently selected
from the group consisting of halogen, C.sub.1-C.sub.6-alkyl,
hydroxy-C.sub.1-C.sub.6-alkyl, C.sub.2-C.sub.6-alkenyl,
C.sub.1-C.sub.6-alkoxy, C.sub.1-C.sub.6-alkoxy-C-
.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-Cycloalkyloxy,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6- -alkoxy,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-
-alkyl, heterocyclyl-C.sub.1-C.sub.6-alkyl, amino, and
amino-C.sub.1-C.sub.6-alkyl, wherein: the nitrogen of the amino or
amino-C.sub.1-C.sub.6-alkyl optionally is substituted with up to
two substituents independently selected from the group consisting
of C.sub.1-C.sub.6-alkyl.
88. A method according to claim 84, wherein the compound inhibits
matrix metalloprotease-13 selectively over both matrix
metalloprotease-1 and matrix metalloprotease-14.
89-90 (canceled).
91. A method for treating a pathological condition in an animal,
wherein: the method comprises administering a compound recited in
claim 14 (or a pharmaceutically-acceptable salt thereof) to the
animal in an amount effective to treat the condition, and the
condition is treatable by inhibiting TNF-.alpha. convertase
activity.
92-95 (canceled).
96. A method for treating a pathological condition in an animal,
wherein: the condition is treatable by inhibiting aggrecanase
activity; and the method comprises administering a compound of
claim 14 (or a pharmaceutically-acceptable salt thereof) to the
animal in an amount effective to treat the condition.
97. A method according to claim 96, herein the compound corresponds
in structure to Formula XA: 608
98. A method according to claim 96, wherein Y is selected from the
group consisting of heterocyclyl, aryl, heteroaryl, and arylmethyl,
wherein: any such substituent optionally is substituted with one or
more substituents independently selected from the group consisting
of halogen, hydroxy, C.sub.1-C.sub.6-alkyl,
halo-C.sub.1-C.sub.6-alkyl, hydroxy-C.sub.1-C.sub.6-alkyl,
C.sub.2-C.sub.6-alkenyl, C.sub.1-C.sub.6-alkoxy,
C.sub.1-C.sub.6-alkylcarbonyl, halo-C.sub.1-C.sub.6-alkoxy,
C.sub.1-C.sub.6-alkylthio,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxycarbo- nyl-C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyl, aryl, aryl-C.sub.1-C.sub.6-alkyl,
aryl-C.sub.1-C.sub.6-alkoxy, heterocyclyl,
heterocyclyl-C.sub.1-C.sub.6-alkyl, heteroaryl, heteroarylcarbonyl,
heterocyclylcarbonyl-C.sub.1-C.sub.6-alkyl, amino, and
amino-C.sub.1-C.sub.6-alkyl, wherein: any such substituent
optionally is substituted with one or more substituents
independently selected from the group consisting of halogen, nitro,
C.sub.1-C.sub.6-alkyl, halo-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxy, and C.sub.1-C.sub.6-alkylcarbonyl, and the
nitrogen of the amino or amino-C.sub.1-C.sub.6-alkyl optionally is
substituted with up to two substituents independently selected from
the group consisting of C.sub.1-C.sub.6-alkyl and
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alky- l.
99. A method according to claim 96, wherein Y is selected from the
group consisting of aryl, heteroaryl, arylmethyl, and
heteroarylmethyl, wherein: any such substituent optionally is
substituted with one or more substituents independently selected
from the group consisting of halogen, C.sub.1-C.sub.6-alkyl,
hydroxy-C.sub.1-C.sub.6-alkyl, C.sub.2-C.sub.6-alkenyl,
C.sub.1-C.sub.6-alkoxy, C.sub.1-C.sub.6-alkoxy-C-
.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyloxy,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6- -alkoxy,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-
-alkyl, heterocyclyl-C.sub.1-C.sub.6-alkyl, amino, and
amino-C.sub.1-C.sub.6-alkyl, wherein: the nitrogen of the amino or
amino-C.sub.1-C.sub.6-alkyl optionally is substituted with up to
two substituents independently selected from the group consisting
of C.sub.1-C.sub.6-alkyl.
100-104 (canceled).
105. A pharmaceutical composition comprising a compound recited in
claim 1 or a pharmaceutically acceptable salt thereof.
106. A pharmaceutical composition according to claim 105, wherein
the compound corresponds in structure to Formula XA: 609
107. A pharmaceutical composition comprising a compound recited in
claim 14 or a pharmaceutically acceptable salt thereof.
108. A pharmaceutical composition according to claim 107, wherein
the compound corresponds in structure to Formula XA: 610
109. A pharmaceutical composition according to claim 107, wherein Y
is selected from the group consisting of heterocyclyl, aryl,
heteroaryl, and arylmethyl, wherein: any such substituent
optionally is substituted with one or more substituents
independently selected from the group consisting of halogen,
hydroxy, C.sub.1-C.sub.6-alkyl, halo-C.sub.1-C.sub.6-alkyl,
hydroxy-C.sub.1-C.sub.6-alkyl, C.sub.2-C.sub.6-alkenyl,
C.sub.1-C.sub.6-alkoxy, C.sub.1-C.sub.6-alkylcarbonyl,
halo-C.sub.1-C.sub.6-alkoxy, C.sub.1-C.sub.6-alkylthio,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxycarbo- nyl-C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyl, aryl, aryl-C.sub.1-C.sub.6-alkyl,
aryl-C.sub.1-C.sub.6-alkoxy, heterocyclyl,
heterocyclyl-C.sub.1-C.sub.6-alkyl, heteroaryl, heteroarylcarbonyl,
heterocyclylcarbonyl-C.sub.1-C.sub.6-alkyl, amino, and
amino-C.sub.1-C.sub.6-alkyl, wherein: any such substituent
optionally is substituted with one or more substituents
independently selected from the group consisting of halogen, nitro,
C.sub.1-C.sub.6-alkyl, halo-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxy, and C.sub.1-C.sub.6-alkylcarbonyl, and the
nitrogen of the amino or amino-C.sub.1-C.sub.6-alkyl optionally is
substituted with up to two substituents independently selected from
the group consisting of C.sub.1-C.sub.6-alkyl and
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alky- l.
110. A pharmaceutical composition according to claim 107, wherein Y
is selected from the group consisting of aryl, heteroaryl,
arylmethyl, and heteroarylmethyl, wherein: any such substituent
optionally is substituted with one or more substituents
independently selected from the group consisting of halogen,
C.sub.1-C.sub.6-alkyl, hydroxy-C.sub.1-C.sub.6-alk- yl,
C.sub.2-C.sub.6-alkenyl, C.sub.1-C.sub.6-alkoxy,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyl-- C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyloxy,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alkoxy,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
heterocyclyl-C.sub.1-C.sub.6-alkyl, amino, and
amino-C.sub.1-C.sub.6-alky- l, wherein: the nitrogen of the amino
or amino-C.sub.1-C.sub.6-alkyl optionally is substituted with up to
two substituents independently selected from the group consisting
of C.sub.1-C.sub.6-alkyl.
111. A compound or salt according to claim 14, wherein the compound
corresponds in structure to the following formula: 611
112. A method according to claim 59, wherein the condition is
osteoarthritis.
113. A method according to claim 79, wherein the condition is
osteoarthritis.
Description
CROSS-REFERENCE TO RELATED PATENT APPLICATIONS
[0001] This patent claims priority as a continuation-in-part to
U.S. patent application Ser. No. 09/570,731 (filed May 12, 2000),
which, in turn, claims priority to U.S. patent application Ser. No.
09/311,837 (filed May 14, 1999) and Ser. No. 09/256,948 (filed Feb.
24, 1999), which, in turn, claim priority to U.S. patent
application Ser. No. 09/191,129 (filed Nov. 13, 1998), Ser. No.
09/186,410 (filed Nov. 5, 1998), 60/066,007 (filed Nov. 14, 1997),
60/095,347 (filed Aug. 4, 1998), 60/095,501 (filed Aug. 6, 1998),
and 60/101,080 (filed Sep. 18, 1998). The entire texts of the
above-referenced patent applications are incorporated by reference
into this patent.
FIELD OF THE INVENTION
[0002] This invention is directed generally to proteinase (also
known as "protease") inhibitors, and, more particularly, to
aromatic sulfone hydroxamate compounds (also known as "aromatic
sulfone hydroxamic acid compounds") and salts thereof (particularly
pharmaceutically acceptable salts) that, inter alia, inhibit matrix
metalloproteinase (also known as "matrix metalloprotease" or "MMP")
and/or aggrecanase activity. This invention also is directed to
pharmaceutical compositions of such compounds and salts, and
methods of using such compounds and salts to prevent or treat
conditions associated with MMP and/or aggrecanase activity,
particularly pathological conditions.
BACKGROUND OF THE INVENTION
[0003] Connective tissue is a required component of all mammals. It
provides rigidity, differentiation, attachments, and, in some
cases, elasticity. Connective tissue components include, for
example, collagen, elastin, proteoglycans, fibronectin, and
laminin. These biochemicals make up (or are components of)
structures, such as skin, bone, teeth, tendon, cartilage, basement
membrane, blood vessels, cornea, and vitreous humor.
[0004] Under normal conditions, connective tissue turnover and/or
repair processes are in equilibrium with connective tissue
production. Degradation of connective tissue is carried out by the
action of proteinases released from resident tissue cells and/or
invading inflammatory or tumor cells.
[0005] Matrix metalloproteinases, a family of zinc-dependent
proteinases, make up a major class of enzymes involved in degrading
connective tissue. Matrix metalloproteinases are divided into
classes, with some members having several different names in common
use. Examples are: MMP-1 (also known as collagenase 1, fibroblast
collagenase, or EC 3.4.24.3); MMP-2 (also known as gelatinase A, 72
kDa gelatinase, basement membrane collagenase, or EC 3.4.24.24),
MMP-3 (also known as stromelysin 1 or EC 3.4.24.17),
proteoglycanase, MMP-7 (also known as matrilysin), MMP-8 (also
known as collagenase II, neutrophil collagenase, or EC 3.4.24.34),
MMP-9 (also known as gelatinase B, 92 kDa gelatinase, or EC
3.4.24.35), MMP-10 (also known as stromelysin 2 or EC 3.4.24.22),
MMP-1 I (also known as stromelysin 3), MMP-12 (also known as
metalloelastase, human macrophage elastase or HME), MMP-13 (also
known as collagenase 111), and MMP-14 (also known as MT1-MMP or
membrane MMP). See, generally, Woessner, J. F., "The Matrix
Metalloprotease Family" in Matrix Metalloproteinases, pp.1-14
(Edited by Parks, W. C. & Mecham, R. P., Academic Press, San
Diego, Calif. 1998).
[0006] Excessive breakdown of connective tissue by MMPs is a
feature of many pathological conditions. Inhibition of MMPs
therefore provides a control mechanism for tissue decomposition to
prevent and/or treat these pathological conditions. Such
pathological conditions generally include, for example, tissue
destruction, fibrotic diseases, pathological matrix weakening,
defective injury repair, cardiovascular diseases, pulmonary
diseases, kidney diseases, liver diseases, and diseases of the
central nervous system. Specific examples of such conditions
include, for example, rheumatoid arthritis, osteoarthritis, septic
arthritis, multiple sclerosis, a decubitis ulcer, corneal
ulceration, epidermal ulceration, gastric ulceration, tumor
metastasis, tumor invasion, tumor angiogenesis, periodontal
disease, liver cirrhosis, fibrotic lung disease, emphysema,
otosclerosis, atherosclerosis, proteinuria, coronary thrombosis,
dilated cardiomyopathy, congestive heart failure, aortic aneurysm,
epidermolysis bullosa, bone disease, Alzheimer's disease, and
defective injury repair (e.g., weak repairs, adhesions such as
post-surgical adhesions, and scarring).
[0007] Matrix metalloproteinases also are involved in the
biosynthesis of tumor necrosis factors (TNFs). Tumor necrosis
factors are implicated in many pathological conditions.
TNF-.alpha., for example, is a cytokine that is presently thought
to be produced initially as a 28 kD cell-associated molecule. It is
released as an active, 17 kD form that can mediate a large number
of deleterious effects in vitro and in vivo. TNF-.alpha. can cause
and/or contribute to the effects of inflammation (e.g. rheumatoid
arthritis), autoimmune disease, graft rejection, multiple
sclerosis, fibrotic diseases, cancer, infectious diseases (e.g.,
malaria, mycobacterial infection, meningitis, etc.), fever,
psoriasis, cardiovascular diseases (e.g., post-ischemic reperfusion
injury and congestive heart failure), pulmonary diseases,
hemorrhage, coagulation, hyperoxic alveolar injury, radiation
damage, and acute phase responses like those seen with infections
and sepsis and during shock (e.g., septic shock and hemodynamic
shock). Chronic release of active TNF-.alpha. can cause cachexia
and anorexia. TNF-.alpha. also can be lethal.
[0008] Inhibiting TNF (and related compounds) production and action
is an important clinical disease treatment. Matrix
metalloproteinase inhibition is one mechanism that can be used. MMP
(e.g., collagenase, stromelysin, and gelatinase) inhibitors, for
example, have been reported to inhibit TNF-.alpha. release. See,
e.g., Gearing et al. Nature 376, 555-557 (1994). See also, McGeehan
et al. See also, Nature 376, 558-561 (1994). MMP inhibitors also
have been reported to inhibit TNF-.alpha. convertase, a
metalloproteinase involved in forming active TNF-.alpha.. See,
e.g., WIPO Int'l Pub. No. WO 94/24140. See also, WIPO Int'l Pub.
No. WO 94/02466. See also, WIPO Int'l Pub. No. WO 97/20824.
[0009] Matrix metalloproteinases also are involved in other
biochemical processes in mammals. These include control of
ovulation, postpartum uterine involution, possibly implantation,
cleavage of APP (.beta.-amyloid precursor protein) to the ainyloid
plaque, and inactivation of (.alpha..sub.1-protease inhibitor
(.alpha..sub.1-PI). Inhibiting MMPs therefore may be a mechanism
that may be used to control of fertility. In addition, increasing
and maintaining the levels of an endogenous or administered serine
protease inhibitor (e.g., .alpha..sub.1-PI) supports the treatment
and prevention of pathological conditions such as emphysema,
pulmonary diseases, inflammatory diseases, and diseases of aging
(e.g., loss of skin or organ stretch and resiliency).
[0010] Numerous metalloproteinase inhibitors are known. See,
generally, Brown, P. D., "Synthetic Inhibitors of Matrix
Metalloproteinases," in Matrix Metalloproteinases, pp. 243-61
(Edited by Parks, W. C. & Mecham, R. P., Academic Press, San
Diego, Calif. 1998).
[0011] Metalloproteinase inhibitors include, for example, natural
biochemicals, such as tissue inhibitor of metalloproteinase (TIMP),
.alpha.2-macroglobulin, and their analogs and derivatives. These
are high-molecular-weight protein molecules that form inactive
complexes with metalloproteinases.
[0012] A number of smaller peptide-like compounds also have been
reported to inhibit metalloproteinases. Mercaptoamide peptidyl
derivatives, for example, have been reported to inhibit angiotensin
coniierting enzyme (also known as ACE) in vitro and in vivo. ACE
aids in the production of angiotensin II, a potent pressor
substance in mammals. Inhibiting ACE leads to lowering of blood
pressure.
[0013] A wide variety of thiol compounds have been reported to
inhibit MMPs. See, e.g., WO95/12389. See also, WO96/11209. See
also, U.S. Pat. No. 4,595,700. See also, U.S. Pat. No.
6,013,649.
[0014] A wide variety of hydroxamate compounds also have been
reported to inhibit MMPs. Such compounds reportedly include
hydroxamates having a carbon backbone. See, e.g., WIPO Int'l Pub.
No. WO 95/29892. See also, WIPO Int'l Pub. No. WO 97/24117. See
also, WIPO Int'l Pub. No. WO 97/49679. See also, European Patent
No. EP 0 780 386. Such compounds also reportedly include
hydroxamates having peptidyl backbones or peptidomimetic backbones.
See, e.g, WIPO Int'l Pub. No. WO 90/05719. See also, WIPO Int'l
Pub. No. WO 93/20047. See also, WIPO Int'l Pub. No. WO 95/09841.
See also, WIPO Int'l Pub. No. WO 96/06074. See also, Schwartz et
al., Progr. Med. Chem., 29:271-334(1992). See also, Rasmussen et
al., PharmacoL Ther., 75(1): 69-75 (1997). See also, Denis et al.,
Invest New Drugs, 15(3): 175-185 (1997). Sulfamato hydroxamates
have additionally been reported to inhibit MMPs. See, WIPO Int'l
Pub. No. WO 00/46221. And various aromatic sulfone hydroxamates
have been reported to inhibit MMPs. See, WIPO Int'l Pub. No. WO
99/25687. See also, WIPO Int'l Pub. No. WO 00/50396. See also, WIPO
Int'l Pub. No. WO 00/69821.
[0015] It is often advantageous for an MMP inhibitor drug to target
a certain MMP(s) over another MMP(s). For example, it is typically
preferred to inhibit MMP-2, MMP-3, MMP-9, and/or MMP-13
(particularly MMP-13) when treating and/or preventing cancer,
inhibiting of metastasis, and inhibiting angiogenesis. It also is
typically preferred to inhibit MMP-13 when preventing and/or
treating osteoarthritis. See, e.g., Mitchell et al., J Clin.
Invest., 97:761-768 (1996). See also, Reboul et al., J Clin.
Invest., 97:2011-2019 (1996). Normally, however, it is preferred to
use a drug that has little or no inhibitory effect on MMP-1 and
MMP-14. This preference stems from the fact that both MMP-1 and
MMP-14 are involved in several homeostatic processes, and
inhibition of MMP-1 and/or MMP-14 consequently tends to interfere
with such processes.
[0016] Many known MMP inhibitors exhibit the same or similar
inhibitory effects against each of the MMPs. For example,
batimastat (a peptidomimetic hydroxamate) has been reported to
exhibit IC.sub.50 values of from about 1 to about 20 nM against
each of MMP-1, MMP-2, MMP-3, MMP-7, and MMP-9. Marimastat (another
peptidomimetic hydroxamate) has been reported to be another
broad-spectrum MMP inhibitor with an enzyme inhibitory spectrum
similar to batimastat, except that Marimastat reportedly exhibited
an IC.sub.50 value against MMP-3 of 230 nM. See Rasmussen et al.,
Pharmacol. Ther., 75(1): 69-75 (1997).
[0017] Meta analysis of data from Phase I/II studies using
Marimastat in patients with advanced, rapidly progressive,
treatment-refractory solid tumor cancers (colorectal, pancreatic,
ovarian, and prostate) indicated a dose-related reduction in the
rise of cancer-specific antigens used as surrogate markers for
biological activity. Although Marimastat exhibited some measure of
efficacy via these markers, toxic side effects reportedly were
observed. The most common drug-related toxicity of Marimastat in
those clinical trials was musculoskeletal pain and stiffness, often
commencing in the small joints in the hands, and then spreading to
the arms and shoulder. A short dosing holiday of 1-3 weeks followed
by dosage reduction reportedly permits treatment to continue. See
Rasmussen et al., Pharmacol. Ther., 75(1): 69-75 (1997). It is
thought that the lack of specificity of inhibitory effect among the
MMPs may be the cause of that effect.
[0018] Another enzyme implicated in pathological conditions
associated with excessive degradation of connective tissue is
aggrecanase, particularly aggrecanase-1 (also known as ADAMTS-4).
Specifically, articular cartilage contains large amounts of the
proteoglycan aggrecan. Proteoglycan aggrecan provides mechanical
properties that help articular cartilage in withstanding
compressive deformation during joint articulation. The loss of
aggrecan fragments and their release into synovial fluid caused by
proteolytic cleavages is a central pathophysiological event in
osteoarthritis and rheumatoid arthritis. It has been reported that
two major cleavage sites exist in the proteolytically sensitive
interglobular domains at the N-terminal region of the aggrecan core
protein. One of those sites has been reported to be cleaved by
several matrix metalloproteases. The other site, however, has been
reported to be cleaved by aggrecanase-1. Thus, inhibiting excessive
aggrecanase activity provides a method for preventing or treating
inflammatory conditions. See generally, Tang, B. L., "ADAMTS: A
Novel Family of Extracellular Matrix Proteases," Int'l Journal of
Biochemistry & Cell Biology, 33, pp. 33-44 (2001). Such
diseases reportedly include, for example, osteoarthritis,
rheumatoid arthritis, joint injury, reactive arthritis, acute
pyrophosphate arthritis, and psoriatic arthritis. See, e.g.,
European Patent Application Publ. No. EP 1 081 137 A1.
[0019] In addition to inflammatory conditions, there also is
evidence that inhibiting aggrecanase may be used for preventing or
treating cancer. For example, excessive levels of aggrecanase-1
reportedly have been observed with a ghoma cell line. It also has
been postulated that the enzymatic nature of aggrecanase and its
similarities with the MMPs would support tumor invasion,
metastasis, and angiogenesis. See Tang, Int'l Journal of
Biochemistry & Cell Biology, 33, pp. 33-44 (2001).
[0020] Various hydroxamate compounds have been reported to inhibit
aggrecanase-1. Such compounds include, for example, those described
in European Patent Application Publ. No. EP 1 081 137 A1. Such
compounds also include, for example, those described in WIPO PCT
Int'l Publ. No. WO 00/09000. Such compounds further include, for
example, those described in WIPO PCT Int'l Publ. No. WO
00/59874.
[0021] In view of the importance of hydroxamate compounds and salts
thereof in the prevention or treatment of several NMP- and/or
aggrecanase-related pathological conditions and the lack of enzyme
specificity exhibited by at least some of the hydroxamates that
have been in clinical trials, there continues to be a need for
hydroxamates having greater enzyme inhibition specificity
(preferably toward MMP-2, MMP-9, MMP-13, and/or aggrecanase, and
particularly toward MMP-13 and/or aggrecanase), while exhibiting
little or no inhibition of MMP activity essential to normal bodily
function (e.g., tissue turnover and repair). The following
disclosure describes hydroxamate compounds and salts thereof that
tend to exhibit such desirable activities.
SUMMARY OF THE INVENTION
[0022] This invention is directed to compounds that inhibit MMP
(particularly MMP-2, MMP-9, and/or MMP-13) and/or aggrecanase
activity, while generally exhibiting relatively little or no
inhibition against MMP activity essential to normal bodily function
(particularly MMP-1 and MMP-14 activity). This invention also is
directed to a method for inhibiting MMP and/or aggrecanase
activity, particularly pathological activity. Such a method is
particularly suitable to be used with mammals, such as humans,
other primates (e.g., monkeys, chimpanzees. etc.), companion
animals (e.g., dogs, cats, horses. etc.), farm animals (e.g.,
goats, sheep, pigs, cattle, etc.), laboratory animals (e.g., mice,
rats, etc.), and wild and zoo animals (e.g., wolves, bears, deer,
etc.).
[0023] Briefly, therefore, the invention is directed in part to a
compound or salt thereof. The compound has a structure
corresponding to Formula X: 1
[0024] The variables Z, R, E, and Y are described in more detail
below.
[0025] The present invention also is directed to treatment methods
that comprise administering a compound described above (or
pharmaceutically-acceptable salt thereof) in an effective amount to
a host mammal having a condition associated with pathological
metalloprotease and/or aggrecanase activity. A contemplated
compound or salt thereof tends to exhibit, for example, inhibitory
activity of one or more matrix metalloprotease (MMP) enzymes (e.g.,
MMP-2, MMP-9 and MMP-13), while exhibiting substantially less
inhibition of MMP-1 and/or MMP-14. By "substantially less" it is
meant that a contemplated compound exhibits an IC.sub.50 value
ratio against one or more of MMP-2, MMP-9, or MMP-13 as compared to
its IC.sub.50 value against MMP-1 and/or MMp-14 (e.g., IC.sub.50
MMP-13:IC.sub.50 MMP-1) that is less than about 1:10, preferably
less than about 1:100, and most preferably less than about 1:1000
in the in vitro inhibition assay described in the Example section
below.
[0026] In one embodiment, the process comprises administering an
above-described compound or pharmaceutically acceptable salt
thereof to the host animal in an amount effective to prevent or
treat the condition. Such a condition may be, for example, tissue
destruction, a fibrotic disease, pathological matrix weakening,
defective injury repair, a cardiovascular disease, a pulmonary
disease, a kidney disease, and a central nervous system disease.
Specific examples of such conditions include osteoarthritis,
rheumatoid arthritis, septic arthritis, tumor invasion, tumor
metastasis, tumor angiogenesis, a decubitis ulcer, a gastric ulcer,
a corneal ulcer, periodontal disease, liver cirrhosis, fibrotic
lung disease, otosclerosis, atherosclerosis, multiple sclerosis,
dilated cardiomyopathy, epidermolysis bullosa, aortic aneurysm,
weak injury repair, an adhesion, scarring, congestive heart
failure, coronary thrombosis, emphysema, proteinuria, and
Alzheimer's disease.
[0027] In another embodiment, the prevention or treatment method
comprises administering an above-described compound or
pharmaceutically acceptable salt thereof to the host animal in an
amount effective to inhibit matrix metalloprotease-2, matrix
metalloprotease-9, and/or matrix metalloprotease-13 activity.
[0028] In another embodiment, the prevention or treatment method
comprises administering an above-described compound or
pharmaceutically acceptable salt, thereof to the host animal in an
amount effective to prevent or treat a condition associated with
TNF-.alpha. convertase activity. Examples of such a condition
include inflammation, a pulmonary disease, a cardiovascular
disease, an autoimmune disease, graft rejection, a fibrotic
disease, cancer, an infectious disease, fever, psoriasis,
hemorrhage, coagulation, radiation damage, acute-phase responses of
shock and sepsis, anorexia, and cachexia.
[0029] In another embodiment, the prevention or treatment method
comprises administering an above-described compound or
pharmaceutically acceptable salt thereof to the host animal in an
amount effective to prevent or treat a condition associated with
aggrecanase activity. Such a condition may be, for example, an
inflammatory disease or cancer.
[0030] This invention additionally is directed, in part, to
pharmaceutical compositions comprising the above-described
compounds or pharmaceutically acceptable salts thereof, and the use
of those compositions in the above-described prevention or
treatment processes.
[0031] This invention further is directed, in part, to the use of
an above-described compound or pharmaceutically acceptable salt
thereof for production of a medicament for use in the treatment of
a condition related to MMP activity. As noted above, such a
condition may be, for example, tissue destruction, a fibrotic
disease, pathological matrix weakening, defective injury repair, a
cardiovascular disease, a pulmonary disease, a kidney disease, and
a central nervous system disease.
[0032] Further benefits of Applicants' invention will be apparent
to one skilled in the art reading this patent.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0033] This detailed description of preferred embodiments is
intended only to acquaint others skilled in the art with
Applicants' invention, its principles, and its practical
application so that others skilled in the art may adapt and apply
the invention in its numerous forms, as they may be best suited to
the requirements of a particular use. This detailed description and
its specific examples, while indicating the preferred embodiments
of this invention, are intended for purposes of illustration only.
This invention, therefore, is not limited to the preferred
embodiments described in this patent, and may be variously
modified.
A. Compounds of this Invention
[0034] In accordance with this invention, Applicants have found
that certain aromatic sulfone hydroxamates tend to be effective
toward inhibiting MMPs, particularly those associated with
excessive (or otherwise pathological) breakdown of connective
tissue. Specifically, Applicants have found that these hydroxamates
tend to be effective for inhibiting MMP-2 MMP-9, and/or MMP-13,
which can be particularly destructive to tissue if present or
generated in abnormally excessive quantities or concentrations.
Applicants also have discovered that many of these hydroxamates
tend to be effective toward inhibiting pathological aggrecanase
activity. Applicants have further discovered that these
hydroxamates tend to be selective toward inhibiting aggrecanase
and/or MMPs associated with pathological condition conditions, and
tend to avoid excessive inhibition of MMPs (particularly MMP-1 and
MMP-14) essential to normal bodily function (e.g., tissue turnover
and repair). Applicants have found, for example, that these
hydroxamates tend to be particularly active toward inhibiting
MMP-2, MMP-9, MMP-13, and/or aggrecanase activity in in vitro
assays that are generally predictive of in vivo activity, while
exhibiting minimal inhibition toward MMP-1 and/or MMP-14 in such
assays. Examples of such in vitro assays are discussed in the
example section below. Compounds (or salts) that are particularly
useful as selective MMP inhibitors exhibit, for example, an in
vitro IC.sub.50 value against one or more of MMP-2, MMP-9, and
MMP-13 that is no greater than about 0.1 times the IC.sub.50 value
against MMP-1 and/or MMP-14, more preferably no greater than about
0.01 times the IC.sub.50 value against MMP-1 and/or MMP-14, and
even more preferably 0.001 times the IC.sub.50 value against MMP-1
and/or MMP-14.
[0035] Without being bound by theory, the advantages of the
selectivity of a contemplated compound can be appreciated by
considering the roles of the various MMP and aggrecanase enzymes.
For example, inhibition of MMP-1 is believed to be undesirable due
to the role of MMP-1 as a housekeeping enzyme (i.e., helping to
maintain normal connective tissue turnover and repair). Inhibition
of MMP-1 can lead to toxicities or side effects such as such as
joint or connective tissue deterioration and pain. On the other
hand, MMP-13 is believed to be intimately involved in the
destruction of joint components in diseases such as osteoarthritis.
Thus, potent and selective inhibition of MMP-13 is typically highly
desirable because such inhibition can have a positive effect on
disease progression in a patient (in addition to having an
anti-inflammatory effect).
[0036] Another advantage of the compounds and salts of this
invention is their tendency to be selective with respect to tumor
necrosis factor release and/or tumor necrosis factor receptor
release. This provides the physician with another factor to help
select the best drug for a particular patient. Without being bound
by theory, it is believed that there are multiple factors to this
type of selectivity to be considered. The first is that presence of
tumor necrosis factor can be desirable for the control of cancer in
the organism, so long as TNF is not present in a toxic excess.
Thus, uncontrolled inhibition of release of TNF can be
counterproductive and actually can be considered an adverse side
effect even in cancer patients. In addition, selectivity with
respect to inhibition of the release of the tumor necrosis factor
receptor can also be desirable. The presence of that receptor can
be desirable for maintaining a controlled tumor necrosis level in
the mammal by binding excess TNF.
[0037] Briefly, therefore, this invention is directed, in part, to
a compound or salt thereof (particularly a pharmaceutically
acceptable salt thereof). The compound has a structure
corresponding to Formula X: 2
[0038] In some preferred embodiments:
[0039] Z is --C(O)--, --N(R.sup.6)--, --O--, --S--, --S(O)--,
--S(O).sub.2--, or --N(S(O).sub.2R.sup.7)--. In some particularly
preferred embodiments, Z is --O--. In other particularly preferred
embodiments, Z is --N(R.sup.6)--.
[0040] R.sup.6 is hydrogen, formyl, sulfonic-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxycarbonyl-C.sub.1-C.sub.6-alkyl,
carboxy-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkylcarbonyl-C.sub.1-C.su- b.6-alkyl,
R.sup.8R.sup.9-aminocarbonyl-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxycarbonyl-C.sub.1-C.sub.6-alkylcarbonyl,
carboxy-C.sub.1-C.sub.6-alkylcarbonyl,
C.sub.1-C.sub.6-alkylcarbonyl-C.su- b.1-C.sub.6-alkylcarbonyl,
C.sub.1-C.sub.6-alkoxycarbonyl, carboxy,
C.sub.1-C.sub.6-alkylcarbonyl, R.sup.8R.sup.9-aminocarbonyl,
aryl-C.sub.1-C.sub.6-alkyl, arylcarbonyl,
bis(C.sub.1-C.sub.6-alkoxy-C.su-
b.1-C.sub.6-alkyl)-C.sub.1-C.sub.6-alkyl, C.sub.1-C.sub.6-alkyl,
halo-C.sub.1-C.sub.6-alkyl, trifluoromethyl-C.sub.1-C.sub.6-alkyl,
perfluoro-C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyl, heteroarylcarbonyl,
heterocyclylcarbonyl, aryl, heterocyclyl, heteroaryl,
C.sub.3-C.sub.8-cycloalkyl-C.sub.1-C.sub.6-alkyl,
aryloxy-C.sub.1-C.sub.6- -alkyl,
heteroaryloxy-C.sub.1-C.sub.6-alkyl, heteroaryl-C.sub.1-C.sub.6-al-
koxy-C.sub.1-C.sub.6-alkyl, heteroarylthio-C.sub.1-C.sub.6-alkyl,
arylsulfonyl, C.sub.1-C.sub.6-alkylsulfonyl,
C.sub.5-C.sub.6-heteroarylsu- lfonyl,
carboxy-C.sub.1-C.sub.6-alkyl, aminocarbonyl,
C.sub.1-C.sub.6-alkylimino(R.sup.10)carbonyl,
arylimino(R.sup.10)carbonyl- ,
C.sub.5-C.sub.6-heterocyclylimino(R.sup.10)carbonyl,
arylthio-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkylthio-C.sub.1-C.sub.6- -alkyl,
arylthio-C.sub.3-C.sub.6-alkenyl, C.sub.1-C.sub.4-alkylthio-C.sub.-
3-C.sub.6-alkenyl,
C.sub.5-C.sub.6-heteroaryl-C.sub.1-C.sub.6-alkyl,
halo-C.sub.1-C.sub.6-alkylcarbonyl,
hydroxy-C.sub.1-C.sub.6-alkylcarbonyl- ,
thiol-C.sub.1-C.sub.6-alkylcarbonyl, C.sub.3-C.sub.6-alkenyl,
C.sub.3-C.sub.6-alkynyl, aryloxycarbonyl,
R.sup.8R.sup.9-aminoimino(R.sup- .10)methyl,
R.sup.8R.sup.9-amino-C.sub.1-C.sub.5-alkylcarbonyl,
hydroxy-C.sub.1-C.sub.5-alkyl, R.sup.8R.sup.9-aminocarbonyl,
R.sup.8R.sup.9-aminocarbonyl-C.sub.1-C.sub.6-alkylcarbonyl,
hydroxyaminocarbonyl, R.sup.8R.sup.9-aminosulfonyl,
R.sup.8R.sup.9-aminosulfonyl-C.sub.1-C.sub.6-alkyl,
R.sup.8R.sup.9-amino-C.sub.1-C.sub.6-alkylsulfonyl, or
R.sup.8R.sup.9-amino-C.sub.1-C.sub.6-alkyl.
[0041] In some particularly preferred embodiments, R.sup.6 is
C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyl,
C.sub.3-C.sub.8-cycloalkyl-C.sub.1-C.sub.6-al- kyl,
C.sub.1-C.sub.6-alkylsulfonyl, C.sub.3-C.sub.6-alkenyl, or
C.sub.3-C.sub.6-alkynyl.
[0042] R.sup.7 is aryl-C.sub.1-C.sub.6-alkyl, aryl, heteroaryl,
heterocyclyl, C.sub.1-C.sub.6-alkyl, C.sub.3-C.sub.6-alkynyl,
C.sub.3-C.sub.6-alkenyl, carboxy-C.sub.1-C.sub.6-alkyl, or
hydroxy-C.sub.1-C.sub.6-alkyl.
[0043] R.sup.8 and R.sup.9 are independently selected from the
group consisting of hydrogen, hydroxy, C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkylcarbonyl, arylcarbonyl, aryl,
aryl-C.sub.1-C.sub.6-alkyl, heteroaryl,
heteroaryl-C.sub.1-C.sub.6-alkyl, C.sub.2-C.sub.6-alkynyl,
C.sub.2-C.sub.6-alkenyl, thiol-C.sub.1-C.sub.6-a- lkyl,
C.sub.1-C.sub.6-alkylthio-C.sub.1-C.sub.6-alkyl, cycloalkyl,
cycloalkyl-C.sub.1-C.sub.6-alkyl,
heterocyclyl-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
aryl-C.sub.1-C.sub.6-alkoxy- -C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkoxy-C.su-
b.1-C.sub.6-alkyl, hydroxy-C.sub.1-C.sub.6-alkyl,
carboxy-C.sub.1-C.sub.6-- alkyl, carboxyaryl-C.sub.1-C.sub.6-alkyl,
aminocarbonyl-C.sub.1-C.sub.6-al- kyl,
aryloxy-C.sub.1-C.sub.6-alkyl, heteroaryloxy-C.sub.1-C.sub.6-alkyl,
arylthio-C.sub.1-C.sub.6-alkyl,
heteroarylthio-C.sub.1-C.sub.6-alkyl, a sulfoxide of any said thio
substituents, a sulfone of any said thio substituents,
trifluoromethyl-C.sub.1-C.sub.6-alkyl, halo-C.sub.1-C.sub.6-alkyl,
alkoxycarbonylamino-C.sub.1-C.sub.6-alkyl, and
amino-C.sub.1-C.sub.6-alkyl. Here, the amino-C.sub.1-C.sub.6-alkyl
nitrogen optionally is substituted with up to 2 substituents
independently selected from the group consisting of
C.sub.1-C.sub.6-alkyl, aryl-C.sub.1-C.sub.6-alkyl, cycloalkyl, and
C.sub.1-C.sub.6-alkylcarbonyl. Preferably, no greater than one of
R.sup.8 and R.sup.9 is hydroxy.
[0044] Alternatively, R.sup.8 and R.sup.9, together with the atom
to which they are bonded, form a 5- to 8-membered heterocyclic or
heteroaryl ring containing up to 2 heteroatoms independently
selected from the group consisting of nitrogen, oxygen, and
sulfur.
[0045] R.sup.10 is hydrogen, hydroxy, C.sub.1-C.sub.6-alkyl, aryl,
aryl-C.sub.1-C.sub.6-alkyl, heteroaryl,
heteroaryl-C.sub.1-C.sub.6-alkyl, C.sub.2-C.sub.6-alkynyl,
C.sub.2-C.sub.6-alkenyl, thiol-C.sub.1-C.sub.6-a- lkyl,
C.sub.1-C.sub.6-alkylthio-C.sub.1-C.sub.6-alkyl, cycloalkyl,
cycloalkyl-C.sub.1-C.sub.6-alkyl,
heterocyclyl-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
aryl-C.sub.1-C.sub.6-alkoxy- -C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkoxy-C.su-
b.1-C.sub.6-alkyl, hydroxy-C.sub.1-C.sub.6-alkyl,
carboxy-C.sub.1-C.sub.6-- alkyl, carboxyaryl-C.sub.1-C.sub.6-alkyl,
aminocarbonyl-C.sub.1-C.sub.6-al- kyl,
aryloxy-C.sub.1-C.sub.6-alkyl, heteroaryloxy-C.sub.1-C.sub.6-alkyl,
arylthio-C.sub.1-C.sub.6-alkyl,
heteroarylthio-C.sub.1-C.sub.6-alkyl, a sulfoxide of any said thio
substituents, a sulfone of any said thio substituents,
trifluoromethyl-C.sub.1-C.sub.6-alkyl, halo-C.sub.1-C.sub.6-alkyl,
alkoxycarbonylamino-C.sub.1-C.sub.6-alkyl, and
amino-C.sub.1-C.sub.6-alkyl. Here, the amino-C.sub.1-C.sub.6-alkyl
nitrogen optionally is substituted with up to 2 substituents
independently selected from the group consisting of
C.sub.1-C.sub.6-alkyl, aryl-C.sub.1-C.sub.6-alkyl, cycloalkyl, and
C.sub.1-C.sub.6-alkylcarbonyl.
[0046] E is a bond, --C(O)--, or --S--.
[0047] Y is hydrogen, alkyl, alkoxy, haloalkyl, aryl, arylalkyl,
cycloalkyl, heteroaryl, hydroxy, aryloxy, arylalkoxy,
heteroaryloxy, heteroarylalkyl, perfluoroalkoxy,
perfluoroalkylthio, trifluoromethylalkyl, alkenyl, heterocyclyl,
cycloalkyl, trifluoromethyl, alkoxycarbonyl, or aminoalkyl. Here,
the aryl, heteroaryl, arylalkyl, or heterocyclyl optionally is
substituted with up to 2 substituents independently selected from
the group consisting of alkylcarbonyl, halo, nitro, arylalkyl,
aryl, alkoxy, trifluoroalkyl, trifluoroalkoxy, and amino. The
amino, in turn, optionally is substituted with up to 2 substituents
independently selected from the group consisting of alkyl and
arylalkyl. In some particularly preferred embodiments, Y comprises
a cyclic structure, i.e., Y is optionally substituted aryl,
arylalkyl, cycloalkyl, heteroaryl, aryloxy, arylalkoxy,
heteroaryloxy, heteroarylalkyl, heterocyclyl, or cycloalkyl. In one
such embodiment, Y is optionally substituted phenyl. In another
such embodiment, Y is optionally substituted phenylmethyl. In still
another such embodiment, Y is optionally substituted heteraryl. And
in still yet another such embodiment, Y is optionally substituted
heteroarylmethyl.
[0048] R is hydrogen, cyano, perfluoroalkyl, trifluoromethoxy,
trifluoromethylthio, haloalkyl, trifluoromethylalkyl,
arylalkoxycarbonyl, aryloxycarbonyl, hydroxy, halo, alkyl, alkoxy,
nitro, thiol, hydroxycarbonyl, aryloxy, arylthio, arylalkyl, aryl,
arylcarbonylamino, heteroaryloxy, heteroarylthio, heteroarylalkyl,
cycloalkyl, heterocylyloxy, heterocylylthio, heterocylylamino,
cycloalkyloxy, cycloalkylthio, heteroarylalkoxy,
heteroarylalkylthio, arylalkoxy, arylalkylthio, arylalkylamino,
heterocylyl, heteroaryl, arylazo, hydroxycarbonylalkoxy,
alkoxycarbonylalkoxy, alkylcarbonyl, arylcarbonyl,
arylalkylcarbonyl, alkylcarbonyloxy, arylalkylcarbonyloxy,
hydroxyalkyl, hydroxyalkoxy, alkylthio, alkoxyalkylthio,
alkoxycarbonyl, aryloxyalkoxyaryl, arylthioalkylthioaryl,
aryloxyalkylthioaryl, arylthioalkoxyaryl, hydroxycarbonylalkoxy,
hydroxycarbonylalkylthio, alkoxycarbonylalkoxy,
alkoxycarbonylalkylthio, amino, aminocarbonyl, or aminoalkyl.
[0049] The nitrogen of an R amino may be unsubstituted.
Alternatively, the amino nitrogen may be substituted with up two
substituents that are independently selected from the group
consisting of alkyl, aryl, heteroaryl, arylalkyl, cycloalkyl,
arylalkoxycarbonyl, alkoxycarbonyl, arylcarbonyl,
arylalkylcarbonyl, heteroarylcarbonyl, heteroarylalkylcarbonyl, and
alkylcarbonyl. Alternatively, the amino nitrogen optionally may be
substituted with two substituents such that the two substituents,
together with the amino nitrogen, form a 5- to 8-member
heterocyclyl or heteroaryl ring that: (i) contains from zero to two
additional heteroatoms that are independently selected from the
group consisting of nitrogen, oxygen, and sulfur; and (ii)
optionally is substituted with up to two substituents independently
selected from the group consisting of aryl, alkyl, heteroaryl,
arylalkyl, heteroarylalkyl, hydroxy, alkoxy, alkylcarbonyl,
cycloalkyl, heterocylylalkyl, alkoxycarbonyl, hydroxyalkyl,
trifluoromethyl, benzofused heterocylylalkyl, hydroxyalkoxyalkyl,
arylalkoxycarbonyl, hydroxycarbonyl, aryloxycarbonyl, benzofused
heterocylylalkoxy, benzofused cycloalkylcarbonyl,
heterocyclylalkylcarbonyl, and cycloalkylcarbonyl.
[0050] The nitrogen of an R aminocarbonyl is may be unsubstituted.
Alternatively, the aminocarbonyl nitrogen may be the reacted amine
of an amino acid. Alternatively, the aminocarbonyl nitrogen may be
substituted with up to two substituents independently selected from
the group consisting of alkyl, hydroxyalkyl,
hydroxyheteroarylalkyl, cycloalkyl, arylalkyl,
trifluoromethylalkyl, heterocylylalkyl, benzofused
heterocylylalkyl, benzofused cycloalkyl, and N,N-dialkylsubstituted
alkylamino-alkyl. Alternatively, the aminocarbonyl nitrogen may be
substituted with two substituents such that the two substituents,
together with the aminocarbonyl nitrogen, form a 5- to 8-member
heterocyclyl or heteroaryl ring that optionally is substituted with
up to two substituents independently selected from the group
consisting of alkyl, alkoxycarbonyl, nitro, heterocyclylalkyl,
hydroxy, hydroxycarbonyl, aryl, arylalkyl, heteroaralkyl, and
amino. Here, the amino nitrogen, in turn, optionally is substituted
with: (i) two substituents independently selected from the group
consisting of alkyl, aryl, and heteroaryl; or (ii) two substituents
such that the two substituents, together with the amino nitrogen,
form a 5- to 8-member heterocyclyl or heteroaryl ring.
[0051] The nitrogen of an R aminoalkyl may be unsubstituted.
Alternatively, the aminoalkyl nitrogen may be substituted with up
to two substituents independently selected from the group
consisting of alkyl, aryl, arylalkyl, cycloalkyl,
arylalkoxycarbonyl, alkoxycarbonyl, and alkylcarbonyl.
Alternatively, the aminoalkyl nitrogen may be substituted with two
substituents such that the two substituents, together with the
aminoalkyl nitrogen, form a 5- to 8-member heterocyclyl or
heteroaryl ring.
[0052] In one particularly preferred embodiment, R is halogen
(preferably chloro or fluoro, and even more preferably chloro). In
another particularly preferred embodiment, R is hydrogen so that
the compound corresponds in structure to Formula XA: 3
[0053] In other embodiments directed to compounds corresponding in
structure to Formula X:
[0054] Z is --C(O)--, --N(R.sup.6)--, --O--, --S--, or
--S(O).sub.2--. In one particularly preferred embodiment, Z is
--N(R.sup.6)--. In another particularly preferred embodiment, Z is
--O--.
[0055] R.sup.6 is hydrogen, arylalkoxycarbonyl, alkylcarbonyl,
alkyl, alkoxyalkyl, cycloalkyl, heteroarylcarbonyl, heteroaryl,
cycloalkylalkyl, alkylsulfonyl, haloalkylcarbonyl, alkenyl,
alkynyl, and R.sup.8R.sup.9-aminoalkylcarbonyl.
[0056] In some particularly preferred embodiments, R.sup.6 is
hydrogen, aryl-C.sub.1-C.sub.6-alkoxycarbonyl,
C.sub.1-C.sub.6-alkoxycarbonyl, C.sub.1-C.sub.6-alkyl (preferably
isopropyl), C.sub.1-C.sub.6-alkoxy-C.su- b.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyl, heteroaryl, heteroarylcarbonyl,
halo-C.sub.1-C.sub.6-alkylcarbonyl, or
R.sup.8R.sup.9-amino-C.sub.1-C.sub.6-alkylcarbonyl.
[0057] In other particularly preferred embodiments, R.sup.6 is
C.sub.1-C.sub.6-alkyl (preferably ethyl),
C.sub.1-C.sub.6-alkoxy-C.sub.1-- C.sub.6-alkyl (preferably
methoxyethyl), C.sub.3-C.sub.6-cycloalkyl (preferably cyclopropyl),
C.sub.3-C.sub.8-cycloalkyl-C.sub.1-C.sub.6-alky- l (preferably
cyclopropylmethyl), C.sub.3-C.sub.6-alkenyl (preferably
C.sub.3-alkenyl), C.sub.3-C.sub.6-alkynyl (preferably
C.sub.3-alkynyl), or C.sub.1-C.sub.6-alkylsulfonyl (preferably
methylsulfonyl).
[0058] R.sup.8 and R.sup.9 are independently selected from the
group consisting of hydrogen, alkyl, alkoxy, hydroxyalkyl,
alkoxyalkyl, hydroxyalkoxyalkyl, heteroarylalkyl, cycloalkylalkyl,
heterocyclylcarbonyl, haloalkyl, and aminoalkyl. Here, the
aminoalkyl nitrogen optionally is substituted with up to two
substituents independently selected from the group consisting of
alkyl.
[0059] Alternatively, R.sup.8 and R.sup.9, together with the atom
to which they are bonded, form a 5- to 8-membered heterocyclyl or
heteroaryl containing up to 3 (in many instances, no greater than
2) heteroatoms independently selected from the group consisting of
nitrogen, oxygen, and sulfur. Here, any such heterocyclyl or
heteroaryl (particularly heterocyclyl) optionally is substituted
with one or more substituents independently selected from the group
consisting of hydroxy, keto, carboxy, alkoxyalkyl, hydroxyalkyl,
hydroxyalkoxyalkyl, alkoxycarbonylalkyl, heterocyclylalkyl,
alkoxycarbonyl, and aminoalkyl. The aminoalkyl nitrogen, in turn,
optionally is substituted with up to two substituents independently
selected from the group consisting of alkyl.
[0060] E is a bond, --C(O)--, or --S--.
[0061] Y is cycloalkyl, 2,3-dihydroindolyl, heterocyclyl, aryl,
heteroaryl, arylalkyl, or heteroarylalkyl. Here, the cycloalkyl,
2,3-dihydroindolyl, heterocyclyl, aryl, heteroaryl, arylalkyl, or
heteroarylalkyl optionally is substituted with one or more
substituents independently selected from the group consisting of
halogen, hydroxy, keto, alkyl, haloalkyl, hydroxyalkyl, alkenyl,
alkoxy, alkylcarbonyl, haloalkoxy, alkylthio, alkoxyalkyl,
alkoxycarbonylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkyloxy,
cycloalkylalkoxy, cycloalkylalkoxyalkyl, aryl, arylalkyl,
arylalkoxy, heterocyclyl, heterocyclylalkyl, heteroaryl,
heteroarylcarbonyl, heterocyclylcarbonylalkyl, alkylsulfonyl,
amino, aminoalkyl, and aminocarbonyl. These optional substituents,
in turn, optionally are substituted with one or more substituents
independently selected from the group consisting of halogen, nitro,
alkyl, haloalkyl, alkoxy, haloalkoxy, and alkylcarbonyl.
Additionally, the nitrogen of the amino, aminoalkyl, or
aminocarbonyl optionally is substituted with up to two substituents
independently selected from the group consisting of alkyl and
cycloalkylalkyl.
[0062] In some preferred embodiments, E is --C(O)--, and Y is
heterocyclyl, aryl (particularly phenyl), heteroaryl, or arylmethyl
(particularly phenylmethyl). Here, the heterocyclyl, aryl,
heteroaryl, or arylmethyl optionally is substituted with one or
more substituents independently selected from the group consisting
of halogen, hydroxy, C.sub.1-C.sub.6-alkyl,
halo-C.sub.1-C.sub.6-alkyl, hydroxy-C.sub.1-C.sub.6-alkyl,
C.sub.2-C.sub.6-alkenyl, C.sub.1-C.sub.6-alkoxy,
C.sub.1-C.sub.6-alkylcarbonyl, halo-C.sub.1-C.sub.6-alkoxy,
C.sub.1-C.sub.6-alkylthio,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxycarbo- nyl-C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyl, aryl, aryl-C.sub.1-C.sub.6-alkyl,
aryl-C.sub.1-C.sub.6-alkoxy, heterocyclyl,
heterocyclyl-C.sub.1-C.sub.6-alkyl, heteroaryl, heteroarylcarbonyl,
heterocyclylcarbonyl-C.sub.1-C.sub.6-alkyl, amino, and
amino-C.sub.1-C.sub.6-alkyl. These optional substituents, in turn,
are optionally substituted with one or more substituents
independently selected from the group consisting of halogen, nitro,
C.sub.1-C.sub.6-alkyl, halo-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxy, and C.sub.1-C.sub.6-alkylcarbonyl.
Additionally, the nitrogen of the amino or
amino-C.sub.1-C.sub.6-alkyl optionally is substituted with up to
two substituents independently selected from the group consisting
of C.sub.1-C.sub.6-alkyl and C.sub.3-C.sub.6-cycloalkyl--
C.sub.1-C.sub.6-alkyl.
[0063] In other preferred embodiments, E is --C(O)--, and Y is aryl
(particularly phenyl), heteroaryl, arylmethyl (particularly
phenylmethyl), or heteroarylmethyl. The aryl, heteroaryl,
arylmethyl, or heteroarylmethyl optionally is substituted with one
or more substituents independently selected from the group
consisting of halogen, C.sub.1-C.sub.6-alkyl,
hydroxy-C.sub.1-C.sub.6-alkyl, C.sub.2-C.sub.6-alkenyl,
C.sub.1-C.sub.6-alkoxy, C.sub.1-C.sub.6-alkoxy-C-
.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alkyl,
C.sub.3-C.sub.6-cycloalkyloxy,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6- -alkoxy,
C.sub.3-C.sub.6-cycloalkyl-C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-
-alkyl, heterocyclyl-C.sub.1-C.sub.6-alkyl, amino, and
amino-C.sub.1-C.sub.6-alkyl. And the nitrogen of the amino or
amino-C.sub.1-C.sub.6-alkyl optionally is substituted with up to
two substituents independently selected from the group consisting
of C.sub.1-C.sub.6-alkyl. In some such preferred embodiments, Y is
optionally substituted phenyl. Such compounds include, for example:
4
[0064] In other such preferred embodiments, Y is optionally
substituted heteroaryl. Such compounds include, for example,
compounds wherein Y is optionally substituted thienyl: 5
[0065] In other preferred embodiments, E is a bond, and Y is aryl
(particularly phenyl), 2,3-dihydroindolyl, heterocyclyl, or
heteroaryl. The aryl, 2,3-dihydroindolyl, heterocyclyl, or
heteroaryl optionally is substituted with one or more substituents
independently selected from the group consisting of halogen, keto,
hydroxy, C.sub.1-C.sub.6-alkyl, C.sub.1-C.sub.6-alkoxy,
halo-C.sub.1-C.sub.6-alkyl, halo-C.sub.1-C.sub.6-alkoxy, aryl,
aminocarbonyl, and C.sub.1-C.sub.6-alkylsulfonyl. These optional
substituents, in turn, also are optionally substituted with one or
more substituents independently selected from the group consisting
of halogen, halo-C.sub.1-C.sub.6-alkyl- , and
halo-C.sub.1-C.sub.6-alkoxy. Additionally, the nitrogen of the
aminocarbonyl optionally is substituted with up to 2 substituents
independently selected from the group consisting of
C.sub.1-C.sub.6-alkyl.
[0066] In other preferred embodiments, E is a bond, and Y is
heteroaryl, aryl (particularly phenyl), or heterocyclyl. The
heteroaryl, aryl, or heterocyclyl optionally is substituted with
one or more substituents independently selected from the group
consisting of halogen, C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxy, and aryl. The optional aryl substituent(s),
in turn, optionally is/are substituted with one or more
substituents independently selected from the group consisting of
halo-C.sub.1-C.sub.6-alkyl.
[0067] In other preferred embodiments, E is --S--, and Y is
cycloalkyl, aryl, arylmethyl, or heteroaryl. The cycloalkyl, aryl
(particularly phenyl), arylmethyl (particularly phenylmethyl), or
heteroaryl optionally is substituted with one or more substituents
independently selected from the group consisting of halogen,
halo-C.sub.1-C.sub.6-alkyl, and halo-C.sub.1-C.sub.6-alkoxy.
[0068] In other preferred embodiments, E is --S--, and Y is
heteraryl.
[0069] In the above embodiments, R preferably is halogen
(preferably chloro or fluoro, and even more preferably chloro).
Alternatively, R preferably is hydrogen so that the compound
corresponds in structure to Formula XA (shown above).
B. Preparation of Useful Compounds
[0070] Exemplary chemical transformations that can be useful for
preparing compounds and salts of this invention are described in
detail in, for example, WIPO Int'l Publ. Nos. WO 00/69821
(published Nov. 23, 2000); WO 00/50396 (published Aug. 31, 2000);
and 99/25687 (published May 27, 1999). These references are hereby
incorporated by reference into this patent. The reader also is
referred to the Example section below, which describes the
preparation of numerous compounds and salts of this invention.
C. Salts of the Compounds of this Invention
[0071] The compounds of this invention can be used in the form of
salts derived from inorganic or organic acids. Depending on the
particular compound, a salt of the compound may be advantageous due
to one or more of the salt's physical properties, such as enhanced
pharmaceutical stability in differing temperatures and humidities,
or a desirable solubility in water or oil. In some instances, a
salt of a compound also may be used as an aid in the isolation,
purification, and/or resolution of the compound.
[0072] Where a salt is intended to be administered to a patient (as
opposed to, for example, being used in an in vitro context), the
salt preferably is pharmaceutically acceptable. Pharmaceutically
acceptable salts include salts commonly used to form alkali metal
salts and to form addition salts of free acids or free bases. In
general, these salts typically may be prepared by conventional
means with a compound of this invention by reacting, for example,
the appropriate acid or base with the compound.
[0073] Pharmaceutically-acceptable acid addition salts of the
compounds of this invention may be prepared from an inorganic or
organic acid. Examples of suitable inorganic acids include
hydrochloric, hydrobromic acid, hydroionic, nitric, carbonic,
sulfuric, and phosphoric acid. Suitable organic acids generally
include, for example, aliphatic, cycloaliphatic, aromatic,
araliphatic, heterocyclyl, carboxylic, and sulfonic classes of
organic acids. Specific examples of suitable organic acids include
acetate, trifluoroacetate, formate, propionate, succinate,
glycolate, gluconate, digluconate, lactate, malate, tartaric acid,
citrate, ascorbate, glucuronate, maleate, fumarate, pyruvate,
aspartate, glutamate, benzoate, anthranilic acid, mesylate,
stearate, salicylate, p-hydroxybenzoate, phenylacetate, mandelate,
embonate (pamoate), methanesulfonate, ethanesulfonate,
benzenesulfonate, pantothenate, toluenesulfonate,
2-hydroxyethanesulfonate, sufanilate, cyclohexylaminosulfonate,
algenic acid, b-hydroxybutyric acid, galactarate, galacturonate,
adipate, alginate, bisulfate, butyrate, camphorate,
camphorsulfonate, cyclopentanepropionate, dodecylsulfate,
glycoheptanoate, glycerophosphate, hemisulfate, heptanoate,
hexanoate, nicotinate, 2-naphthalesulfonate, oxalate, palmoate,
pectinate, persulfate, 3-phenylpropionate, picrate, pivalate,
thiocyanate, tosylate, and undecanoate.
[0074] Pharmaceutically-acceptable base addition salts of the
compounds of this invention include, for example, metallic salts
and organic salts. Preferred metallic salts include alkali metal
(group Ia) salts, alkaline earth metal (group IIa) salts, and other
physiological acceptable metal salts. Such salts may be made from
aluminum, calcium, lithium, magnesium, potassium, sodium, and zinc.
Preferred organic salts can be made from tertiary amines and
quaternary amine salts, such as tromethamine, diethylamine,
N,N'-dibenzylethylenediamine, chloroprocaine, choline,
diethanolamine, ethylenediamine, meglumine (N-methylglucamine), and
procaine. Basic nitrogen-containing groups can be quaternized with
agents such as lower alkyl (C.sub.1-C.sub.6) halides (e.g., methyl,
ethyl, propyl, and butyl chlorides, bromides, and iodides), dialkyl
sulfates (e.g., dimethyl, diethyl, dibuytl, and diamyl sulfates),
long chain halides (e.g., decyl, lauryl, myristyl, and stearyl
chlorides, bromides, and iodides), aralkyl halides (e.g., benzyl
and phenethyl bromides), and others.
[0075] Particularly preferred salts of the compounds of this
invention include hydrochloric acid (HCl) salts and
trifluoroacetate (CF.sub.3COOH or TFA) salts.
D. Preventing or Treating Conditions Using the Compounds and Salts
of this Invention
[0076] One embodiment of this invention is directed to a process
for preventing or treating a pathological condition associated with
MMP activity in a mammal (e.g., a human, companion animal, farm
animal, laboratory animal, zoo animal, or wild animal) having or
disposed to having such a condition. Such a condition may be, for
example, tissue destruction, a fibrotic disease, pathological
matrix weakening, defective injury repair, a cardiovascular
disease, a pulmonary disease, a kidney disease, and a central
nervous system disease. Specific examples of such conditions
include osteoarthritis, rheumatoid arthritis, septic arthritis,
tumor invasion, tumor metastasis, tumor angiogenesis, a decubitis
ulcer, a gastric ulcer, a corneal ulcer, periodontal disease, liver
cirrhosis, fibrotic lung disease, otosclerosis, atherosclerosis,
multiple sclerosis, dilated cardiomyopathy, epidermolysis bullosa,
aortic aneurysm, weak injury repair, an adhesion, scarring,
congestive heart failure, coronary thrombosis, emphysema,
proteinuria, and Alzheimer's disease.
[0077] The condition may alternatively (or additionally) be
associated with TNF-.alpha. convertase activity. Examples of such a
condition include inflammation (e.g., rheumatoid arthritis),
autoimmune disease, graft rejection, multiple sclerosis, a fibrotic
disease, cancer, an infectious disease (e.g., malaria,
mycobacterial infection, meningitis, etc.), fever, psoriasis, a
cardiovascular disease (e.g., post-ischemic reperfusion injury and
congestive heart failure), a pulmonary disease, hemorrhage,
coagulation, hyperoxic alveolar injury, radiation damage, acute
phase responses like those seen with infections and sepsis and
during shock (e.g., septic shock, hemodynamic shock, etc.),
cachexia, and anorexia.
[0078] The condition may alternatively (or additionally) be
associated with aggrecanase activity. Examples of such a condition
include inflammation diseases (e.g., osteoarthritis, rheumatoid
arthritis, joint injury, reactive arthritis, acute pyrophosphate
arthritis, and psoriatic arthritis) and cancer.
[0079] In this patent, the phrase "preventing a condition" means
reducing the risk of (or delaying) the onset of the condition in a
mammal that does not have the condition, but is predisposed to
having the condition. In contrast, the phrase "treating a
condition" means ameliorating, suppressing, or eradicating an
existing condition. The pathological condition may be, for example:
(a) the result of pathological MMP and/or aggrecanase activity
itself, (b) affected by MMP activity (e.g., diseases associated
with TNF-.alpha.), and/or (c) affected by aggrecanase activity.
[0080] A wide variety of methods may be used alone or in
combination to administer the hydroxamates and salt thereof
described above. For example, the hydroxamates or salts thereof may
be administered orally, parenterally, by inhalation spray,
rectally, or topically. Oral administration can be advantageous if,
for example, the patient is ambulatory, not hospitalized, and
physically able and sufficiently responsible to take drug at the
required intervals. This may be true even if the person is being
treated with more than one drug for one or more diseases. On the
other hand, IV drug administration can be advantageous in, for
example, a hospital setting where the dose (and thus the blood
levels) can be well controlled. A compound or salt of this
invention also can be formulated for IM administration if desired.
This route of administration may be desirable for administering
prodrugs or regular drug delivery to patients that are either
physically weak or have a poor compliance record or require
constant drug blood levels.
[0081] Typically, a compound (or pharmaceutically acceptable salt
thereof) described in this patent is administered in an amount
effective to inhibit a target MMP(s). The target MMP is/are
typically MMP-2, MMP-9, and/or MMP-13, with MMP-13 often being a
particularly preferred target. The preferred total daily dose of
the hydroxamate or salt thereof (administered in single or divided
doses) is typically from about 0.001 to about 100 mg/kg, more
preferably from about 0.001 to about 30 mg/kg, and even more
preferably from about 0.01 to about 10 mg/kg (i.e., mg hydroxamate
or salt thereof per kg body weight). Dosage unit compositions can
contain such amounts or submultiples thereof to make up the daily
dose. In many instances, the administration of the compound or salt
will be repeated a plurality of times. Multiple doses per day
typically may be used to increase the total daily dose, if
desired.
[0082] Factors affecting the preferred dosage regimen include the
type, age, weight, sex, diet, and condition of the patient; the
severity of the pathological condition; the route of
administration; pharmacological considerations, such as the
activity, efficacy, pharmacokinetic, and toxicology profiles of the
particular hydroxamate or salt thereof employed; whether a drug
delivery system is utilized; and whether the hydroxamate or salt
thereof is administered as part of a drug combination. Thus, the
dosage regimen actually employed can vary widely, and, therefore,
can deviate from the preferred dosage regimen set forth above.
E. Pharmaceutical Compositions Containing the Compounds and Salts
of this Invention
[0083] This invention also is directed to pharmaceutical
compositions comprising a hydroxamate or salt thereof described
above, and to methods for making pharmacetucal compositions (or
medicaments) comprising a hydroxamate or salt thereof described
above.
[0084] The preferred composition depends on the method of
administration, and typically comprises one or more conventional
pharmaceutically acceptable carriers, adjuvants, and/or vehicles.
Formulation of drugs is generally discussed in, for example,
Hoover, John E., Remington's Pharmaceutical Sciences (Mack
Publishing Co., Easton, Pa.: 1975). See also, Liberman, H. A. See
also, Lachman, L., eds., Pharmaceutical Dosage Forms (Marcel
Decker, New York, N.Y., 1980).
[0085] Solid dosage forms for oral administration include, for
example, capsules, tablets, pills, powders, and granules. In such
solid dosage forms, the hydroxamates or salts thereof are
ordinarily combined with one or more adjuvants. If administered per
os, the hydroxamates or salts thereof can be mixed with lactose,
sucrose, starch powder, cellulose esters of alkanoic acids,
cellulose alkyl esters, talc, stearic acid, magnesium stearate,
magnesium oxide, sodium and calcium salts of phosphoric and
sulfuric acids, gelatin, acacia gum, sodium alginate,
polyvinylpyrrolidone, and/or polyvinyl alcohol, and then tableted
or encapsulated for convenient administration. Such capsules or
tablets can contain a controlled-release formulation, as can be
provided in a dispersion of the hydroxamate or salt thereof in
hydroxypropylmethyl cellulose. In the case of capsules, tablets,
and pills, the dosage forms also can comprise buffering agents,
such as sodium citrate, or magnesium or calcium carbonate or
bicarbonate. Tablets and pills additionally can be prepared with
enteric coatings.
[0086] Liquid dosage forms for oral administration include, for
example, pharmaceutically acceptable emulsions, solutions,
suspensions, syrups, and elixirs containing inert diluents commonly
used in the art (e.g., water). Such compositions also can comprise
adjuvants, such as wetting, emulsifying, suspending, flavoring
(e.g., sweetening), and/or perfuming agents.
[0087] "Parenteral administration" includes subcutaneous
injections, intravenous injections, intramuscular injections,
intrasternal injections, and infusion. Injectable preparations
(e.g., sterile injectable aqueous or oleaginous suspensions) can be
formulated according to the known art using suitable dispersing,
wetting agents, and/or suspending agents. Acceptable vehicles and
solvents include, for example, water, 1,3-butanediol, Ringer's
solution, isotonic sodium chloride solution, bland fixed oils
(e.g., synthetic mono- or diglycerides), fatty acids (e.g., oleic
acid), dimethyl acetamide, surfactants (e.g., ionic and non-ionic
detergents), and/or polyethylene glycols.
[0088] Formulations for parenteral administration may, for example,
be prepared from sterile powders or granules having one or more of
the carriers or diluents mentioned for use in the formulations for
oral administration. The hydroxamates or salts thereof can be
dissolved in water, polyethylene glycol, propylene glycol, ethanol,
corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol,
sodium chloride, and/or various buffers.
[0089] Suppositories for rectal administration can be prepared by,
for example, mixing the drug with a suitable nonirritating
excipient that is solid at ordinary temperatures, but liquid at the
rectal temperature and will therefore melt in the rectum to release
the drug. Suitable excipients include, for example, such as cocoa
butter; synthetic mono-, di-, or triglycerides; fatty acids; and/or
polyethylene glycols
[0090] "Topical administration" includes the use of transdermal
administration, such as transdermal patches or iontophoresis
devices.
[0091] Other adjuvants and modes of administration known in the
pharmaceutical art may also be used.
F. Definitions
[0092] The term "alkyl" (alone or in combination with another
term(s)) means a straight-or branched-chain saturated hydrocarbyl
group typically containing from 1 to about 20 carbon atoms, more
typically from about 1 to about 8 carbon atoms, and even more
typically from about 1 to about 6 carbon atoms. Examples of such
groups include methyl, ethyl, n-propyl, isopropyl, n-butyl,
isobutyl, sec-butyl, tert-butyl, pentyl, iso-amyl, hexyl, octyl,
and the like.
[0093] The term "alkenyl" (alone or in combination with another
term(s)) means a straight- or branched-chain hydrocarbyl group
containing one or more double bonds and typically from 2 to about
20 carbon atoms, more typically from about 2 to about 8 carbon
atoms, and even more typically from about 2 to about 6 carbon
atoms. Examples of such groups include ethenyl (vinyl); 2-propenyl;
3-propenyl; 1,4-pentadienyl; 1,4-butadienyl; 1-butenyl; 2-butenyl;
3-butenyl; decenyl; and the like.
[0094] The term "alkynyl" (alone or in combination with another
term(s)) means a straight- or branched-chain hydrocarbyl group
containing one or more triple bonds and typically from 2 to about
20 carbon atoms, more typically from about 2 to about 8 carbon
atoms, and even more typically from about 2 to about 6 carbon
atoms. Examples of such groups include ethynyl, 2-propynyl,
3-propynyl, decynyl, 1-butynyl, 2-butynyl, 3-butynyl, and the
like.
[0095] The term "carbocyclyl" (alone or in combination with another
term(s)) means a saturated cyclic, partially saturated cyclic, or
aryl hydrocarbyl group containing from 3 to 14 carbon ring atoms
("ring atoms" are the atoms bound together to form the ring or
rings of a cyclic group). A carbocyclyl may be a single ring, which
typically contains from 3 to 6 ring atoms. Examples of such
single-ring carbocyclyls include cyclopropanyl, cyclobutanyl,
cyclopentyl, cyclopentenyl, cyclopentadienyl, cyclohexyl,
cyclohexenyl, cyclohexadienyl, and phenyl. A carbocyclyl
alternatively may be 2 or 3 rings fused together, such as
naphthalenyl, tetrahydronaphthalenyl (also known as "tetralinyl"),
indenyl, isoindenyl, indanyl, bicyclodecanyl, anthracenyl,
phenanthrene, benzonaphthenyl (also known as "phenalenyl"),
fluoreneyl, decalinyl, and norpinanyl.
[0096] The term "cycloalkyl" (alone or in combination with another
term(s)) means a saturated cyclic hydrocarbyl group containing from
3 to 14 carbon ring atoms. A cycloalkyl may be a single carbon
ring, which typically contains from 3 to 6 carbon ring atoms.
Examples of single-ring cycloalkyls include cyclopropanyl,
cyclobutanyl, cyclopentyl, and cyclohexyl. A cycloalkyl
alternatively may be 2 or 3 carbon rings fused together, such as,
decalinyl or norpinanyl.
[0097] The term "aryl" (alone or in combination with another
term(s)) means an aromatic carbocyclyl containing from 6 to 14
carbon ring atoms. Examples of aryls include phenyl, naphthalenyl,
and indenyl.
[0098] In some instances, the number of carbon atoms in a
hydrocarbyl group (e.g., alkyl, alkenyl, alkynyl, or cycloalkyl) is
indicated by the prefix "C.sub.x-C.sub.y-", wherein x is the
minimum and y is the maximum number of carbon atoms in the group.
Thus, for example, "C.sub.1-C.sub.6-alkyl" refers to an alkyl group
containing from 1 to 6 carbon atoms. Illustrating further,
C.sub.3-C.sub.6-cycloalkyl means a saturated hydrocarbyl ring
containing from 3 to 6 carbon ring atoms.
[0099] The term "hydrogen" (alone or in combination with another
term(s)) means a hydrogen radical, and may be depicted as --H.
[0100] The term "hydroxy" (alone or in combination with another
term(s)) means --OH.
[0101] The term "nitro" (alone or in combination with another
term(s)) means --NO.sub.2.
[0102] The term "cyano" (alone or in combination with another
term(s)) means --CN, which also may be depicted as or --COOH: 6
[0103] The term "keto" (alone or in combination with another
term(s)) means an oxo radical, and may be depicted as .dbd.O.
[0104] The term "carboxy" (alone or in combination with another
term(s)) means --C(O)--OH, which also may be depicted as: 7
[0105] The term "amino" (alone or in combination with another
term(s)) means --NH.sub.2. The term "monosubstituted amino" (alone
or in combination with another term(s)) means an amino group
wherein one of the hydrogen radicals is replaced by a non-hydrogen
substituent. The term "disubstituted amino" (alone or in
combination with another term(s)) means an amino group wherein both
of the hydrogen atoms are replaced by non-hydrogen substituents,
which may be identical or different.
[0106] The term "halogen" (alone or in combination with another
term(s)) means a fluorine radical (which may be depicted as --F),
chlorine radical (which may be depicted as --Cl), bromine radical
(which may be depicted as --Br), or iodine radical (which may be
depicted as --I). Typically, a fluorine radical or chlorine radical
is preferred.
[0107] If a group is described as being "substituted", at least one
hydrogen on the group is replaced with a non-hydrogen substituent.
Thus, for example, a substituted alkyl group is an alkyl group
wherein at least one hydrogen on the alkyl group is replaced with a
non-hydrogen substituent. It should be recognized that if there are
more than one substitutions on a group, each non-hydrogen
substituent may be identical or different.
[0108] If a group is described as being "optionally substituted",
the group may be either substituted or not substituted.
[0109] The prefix "halo" indicates that the group to which the
prefix is attached is substituted with one or more independently
selected halogen radicals. For example, haloalkyl means an alkyl
group wherein at least one hydrogen radical is replaced with a
halogen radical. Examples of haloalkyls include chloromethyl,
1-bromoethyl, fluoromethyl, difluoromethyl, trifluoromethyl,
1,1,1-trifluoroethyl, and the like. Illustrating further,
"haloalkoxy" means an alkoxy group wherein at least one hydrogen
radical is replaced by a halogen radical. Examples of haloalkoxy
groups include chlormethoxy, 1-bromoethoxy, fluoromethoxy,
difluoromethoxy, trifluoromethoxy (also known as
"perfluoromethyoxy"), 1,1,1,-trifluoroethoxy, and the like. It
should be recognized that if a group is substituted by more than
one halogen radical, those halogen radicals may be identical or
different.
[0110] The prefix "perhalo" indicates that every hydrogen radical
on the group to which the prefix is attached is replaced with
independently selected halogen radicals, i.e., each hydrogen
radical on the group is replaced with a halogen radical. If all the
halogen radicals are identical, the prefix typically will identify
the halogen radical. Thus, for example, the term "perfluoro" means
that every hydrogen radical on the group to which the prefix is
attached is substituted with a fluorine radical. To illustrate, the
term "prefluoroalkyl" means an alkyl group wherein each hydrogen
radical is replaced with a fluorine radical. Examples of
perfluoroalkyl groups include trifluoromethyl (--CF.sub.3),
perfluorobutyl, perfluoroisopropyl, perfluorododecyl,
perfluorodecyl, and the like. To illustrate further, the term
"perfluoroalkoxy" means an alkoxy group wherein each hydrogen
radical is replaced with a fluorine radical. Examples of
perfluoroalkoxy groups include trifluoromethoxy (--O--CF.sub.3),
perfluorobutoxy, perfluoroisopropoxy, perfluorododecoxy,
perfluorodecoxy, and the like.
[0111] The term "carbonyl" (alone or in combination with another
term(s)) means --C(O)--, which also may be depicted as: 8
[0112] This term also is intended to encompass a hydrated carbonyl
group, i.e., --C(OH).sub.2--.
[0113] The term "aminocarbonyl" (alone or in combination with
another term(s)) means --C(O)--NH.sub.2, which also may be depicted
as: 9
[0114] The term "oxy" (alone or in combination with another
term(s)) means an ether group, and may be depicted as --O--.
[0115] The term "alkoxy" (alone or in combination with another
term(s)) means an alkylether group, i.e., --O-alkyl. Examples of
such a group include methoxy (--O--CH.sub.3), ethoxy, n-propoxy,
isopropoxy, n-butoxy, iso-butoxy, sec-butoxy, tert-butoxy, and the
like.
[0116] The term "alkylcarbonyl" (alone or in combination with
another term(s)) means --C(O)-alkyl. For example, "ethylcarbonyl"
may be depicted as: 10
[0117] The term "aminoalkylcarbonyl" (alone or in combination with
another term(s)) means --C(O)-alkyl-NH.sub.2. For example,
"aminomethylcarbonyl" may be depicted as: 11
[0118] The term "alkoxycarbonyl" (alone or in combination with
another term(s)) means --C(O)--O-alkyl. For example,
"ethoxycarbonyl" may be depicted as: 12
[0119] The term "carbocyclylcarbonyl" (alone or in combination with
another term(s)) means --C(O)-carbocyclyl. For example,
"phenylcarbonyl" may be depicted as: 13
[0120] Similarly, the term "heterocyclylcarbonyl" (alone or in
combination with another term(s)) means --C(O)-heterocyclyl.
[0121] The term "carbocyclylalkylcarbonyl" (alone or in combination
with another term(s)) means --C(O)-alkyl-carbocyclyl. For example,
"phenylethylcarbonyl" may be depicted as: 14
[0122] Similarly, the term "heterocyclylalkylcarbonyl" (alone or in
combination with another term(s)) means
--C(O)-alkyl-heterocyclyl.
[0123] The term "carbocyclyloxycarbonyl" (alone or in combination
with another term(s)) means --C(O)--O-carbocyclyl. For example,
"phenyloxycarbonyl" may be depicted as: 15
[0124] The term "carbocyclylalkoxycarbonyl" (alone or in
combination with another term(s)) means
--C(O)--O-alkyl-carbocyclyl. For example, "phenylethoxycarbonyl"
may be depicted as: 16
[0125] The term "thio" or "thia" (alone or in combination with
another term(s)) means a thiaether group, i.e., an ether group
wherein the ether oxygen atom is replaced by a sulfur atom. Such a
group may be depicted as --S--. This, for example,
"alkyl-thio-alkyl" means alkyl-S-alkyl.
[0126] The term "thiol" or "sulfhydryl" (alone or in combination
with another term(s)) means a sulfhydryl group, and may be depicted
as --SH.
[0127] The term "(thiocarbonyl)" (alone or in combination with
another term(s)) means a carbonyl wherein the oxygen atom has been
replaced with a sulfur. Such a group may be depicted as --C(S)--,
and also may be depicted as: 17
[0128] The term "alkyl(thiocarbonyl)" (alone or in combination with
another term(s)) means --C(S)-alkyl. For example,
"ethyl(thiocarbonyl)" may be depicted as: 18
[0129] The term "alkoxy(thiocarbonyl)" (alone or in combination
with another term(s)) means --C(S)--O-alkyl. For example,
"ethoxy(thiocarbonyl)" may be depicted as: 19
[0130] The term "carbocyclyl(thiocarbonyl)" (alone or in
combination with another term(s)) means --C(S)-carbocyclyl. For
example, "phenyl(thiocarbonyl)" may be depicted as: 20
[0131] Similarly, the term "heterocyclyl(thiocarbonyl)" (alone or
in combination with another term(s)) means --C(S)-heterocyclyl.
[0132] The term "carbocyclylalkyl(thiocarbonyl)" (alone or in
combination with another term(s)) means --C(S)-alkyl-carbocyclyl.
For example, "phenylethyl(thiocarbonyl)" may be depicted as: 21
[0133] Similarly, the term "heterocyclylalkyl(thiocarbonyl)" (alone
or in combination with another term(s)) means
--C(S)-alkyl-heterocyclyl.
[0134] The term "carbocyclyloxy(thiocarbonyl)" (alone or in
combination with another term(s)) means --C(S)--O-carbocyclyl. For
example, "phenyloxy(thiocarbonyl)" may be depicted as: 22
[0135] The term "carbocyclylalkoxy(thiocarbonyl)" (alone or in
combination with another term(s)) means
--C(S)--O-alkyl-carbocyclyl. For example,
"phenylethoxy(thiocarbonyl)" may be depicted as: 23
[0136] The term "sulfonyl" (alone or in combination with another
term(s)) means --S(O).sub.2--, which also may be depicted as:
24
[0137] Thus, for example, "alkyl-sulfonyl-alkyl" means
alkyl-S(O).sub.2-alkyl.
[0138] The term "aminosulfonyl" (alone or in combination with
another term(s)) means --S(O).sub.2--NH.sub.2, which also may be
depicted as: 25
[0139] The term "sulfoxido" (alone or in combination with another
term(s)) means --S(O)--, which also may be depicted as: 26
[0140] Thus, for example, "alkyl-sulfoxido-alkyl" means
alkyl-S(O)-alkyl.
[0141] The term "heterocyclyl" (alone or in combination with
another term(s)) means a saturated or partially saturated ring
structure containing a total of 3 to 14 ring atoms. At least one of
the ring atoms is a heteroatom (i.e., oxygen, nitrogen, or sulfur),
with the remaining ring atoms being independently selected from the
group consisting of carbon, oxygen, nitrogen, and sulfur. A
heterocyclyl may be a single ring, which typically contains from 3
to 7 ring atoms, more typically from 3 to 6 ring atoms, and even
more typically 5 to 6 ring atoms. A heterocyclyl alternatively may
be 2 or 3 rings fused together.
[0142] The term "heteroaryl" (alone or in combination with another
term(s)) means an aromatic ring containing from 5 to 14 ring atoms.
At least one of the ring atoms is a heteroatom, with the remaining
ring atoms being independently selected from the group consisting
of carbon, oxygen, nitrogen, and sulfur. A heteroaryl may be a
single ring, which typically contains from 5 to 7 ring atoms, and
more typically from 5 to 6 ring atoms. A heteroaryl alternatively
may be 2 or 3 rings fused together.
[0143] Examples of single-ring heterocyclyls and heteroaryls
include furanyl, dihydrofurnayl, tetradydrofurnayl, thiophenyl
(also known as "thiofuranyl"), dihydrothiophenyl,
tetrahydrothiophenyl, pyrrolyl, isopyrrolyl, pyrrolinyl,
pyrrolidinyl, imidazolyl, isoimidazolyl, imidazolinyl,
imidazolidinyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, triazolyl,
tetrazolyl, dithiolyl, oxathiolyl, oxazolyl, isoxazolyl, thiazolyl,
isothiazolyl, thiazolinyl, isothiazolinyl, thiazolidinyl,
isothiazolidinyl, thiodiazolyl, oxathiazolyl, oxadiazolyl
(including 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl (also known as
"azoximyl"), 1,2,5-oxadiazolyl (also known as "furazanyl"), or
1,3,4-oxadiazolyl), oxatriazolyl (including 1,2,3,4-oxatriazolyl or
1,2,3,5-oxatriazolyl), dioxazolyl (including 1,2,3-dioxazolyl,
1,2,4-dioxazolyl, 1,3,2-dioxazolyl, or 1,3,4-dioxazolyl),
oxathiazolyl, oxathiolyl, oxathiolanyl, pyranyl (including
1,2-pyranyl or 1,4-pyranyl), dihydropyranyl, pyridinyl (also known
as "azinyl"), piperidinyl, diazinyl (including pyridazinyl (also
known as "1,2-diazinyl"), pyrimidinyl (also known as
"1,3-diazinyl"), or pyrazinyl (also known as "1,4-diazinyl")),
piperazinyl, triazinyl (including s-triazinyl (also known as
"1,3,5-triazinyl"), as-triazinyl (also known 1,2,4-triazinyl), and
v-triazinyl (also known as "1,2,3-triazinyl")), oxazinyl (including
1,2,3-oxazinyl, 1,3,2-oxazinyl, 1,3,6-oxazinyl (also known as
"pentoxazolyl"), 1,2,6-oxazinyl, or 1,4-oxazinyl), isoxazinyl
(including o-isoxazinyl or p-isoxazinyl), oxazolidinyl,
isoxazolidinyl, oxathiazinyl (including 1,2,5-oxathiazinyl or
1,2,6-oxathiazinyl), oxadiazinyl (including 1,4,2-oxadiazinyl or
1,3,5,2-oxadiazinyl), morpholinyl, azepinyl, oxepinyl, thiepinyl,
and diazepinyl.
[0144] Examples of heterocyclyl and heteroaryl rings having 2 or 3
rings fused together include, for example, indolizinyl, pyrindinyl,
pyranopyrrolyl, 4H-quinolizinyl, purinyl, naphthyridinyl,
pyridopyridinyl (including pyrido[3,4-b]-pyridinyl,
pyrido[3,2-b]-pyridinyl, or pyrido[4,3-b]-pyridinyl), and
pteridinyl. Other examples of fused-ring heterocyclyls include
benzo-fused heterocyclyls, such as indolyl, isoindolyl (also known
as "isobenzazolyl" or "pseudoisoindolyl"), indoleninyl (also known
as "pseudoindolyl"), isoindazolyl (also known as "benzpyrazolyl"),
benzazinyl (including quinolinyl (also known as "1-benzazinyl") or
isoquinolinyl (also known as "2-benzazinyl")), phthalazinyl,
quinoxalinyl, quinazolinyl, benzodiazinyl (including cinnolinyl
(also known as "1,2-benzodiazinyl") or quinazolinyl (also known as
"1,3-benzodiazinyl")), benzopyranyl (including "chromanyl" or
"isochromanyl"), benzothiopyranyl (also known as "thiochromanyl"),
benzoxazolyl, indoxazinyl (also known as "benzisoxazolyl"),
anthranilyl, benzodioxolyl, benzodioxanyl, benzoxadiazolyl,
benzofuranyl (also known as "coumaronyl"), isobenzofuranyl,
benzothienyl (also known as "benzothiophenyl", "thionaphthenyl", or
"benzothiofuiranyl"), isobenzothienyl (also known as
"isobenzothiophenyl", "isothionaphthenyl", or
"isobenzothiofuiranyl"), benzothiazolyl, benzothiadiazolyl,
benzimidazolyl, benzotriazolyl, benzoxazinyl (including
1,3,2-benzoxazinyl, 1,4,2-benzoxazinyl, 2,3,1-benzoxazinyl, or
3,1,4-benzoxazinyl), benzisoxazinyl (including 1,2-benzisoxazinyl
or 1,4-benzisoxazinyl), tetrahydroisoquinolinyl, carbazolyl,
xanthenyl, and acridinyl.
[0145] As may be seen in the preceding paragraphs, the term
"heteroaryl" includes 6-membered ring substituents such as pyridyl,
pyrazyl, pyrimidinyl, and pyridazinyl; 5-membered ring substituents
such as 1,3,5-, 1,2,4- or 1,2,3-tiiazinyl, imidazyl, furanyl,
thiophenyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, 1,2,3-,
1,2,4-, 1,2,5-, or 1,3,4-oxadiazolyl and isothiazolyl; 6/5-membered
fused ring substituents such as benzothiofuranyl,
isobenzothiofuranyl, benzisoxazolyl, benzoxazolyl, purinyl, and
anthranilyl; and 6/6-membered fused rings such as 1,2-, 1,4-, 2,3-
and 2,1-benzopyronyl, quinolinyl, isoquinolinyl, cinnolinyl,
quinazolinyl, and 1,4-benzoxazinyl.
[0146] A carbocyclyl, heterocyclyl, or heteroaryl optionally can be
substituted with, for example, one or more substituents
independently selected from the group consisting of halogen,
hydroxy, carboxy, keto, alkyl, alkoxy, alkoxyalkyl, alkylcarbonyl
(also known as "alkanoyl"), aryl, arylalkyl, arylalkoxy,
arylalkoxyalkyl, arylalkoxycarbonyl, cycloalkyl, cycloalkylalkyl,
cycloalkylalkoxy, cycloalkylalkoxyalkyl, and
cycloalkylalkoxycarbonyl. More typically, a carbocyclyl or
heterocyclyl may optionally be substituted with, for example, one
or more substituents independently selected from the group
consisting of halogen, --OH, --C(O)--OH, keto,
C.sub.1-C.sub.6-alkyl, C.sub.1-C.sub.6-alkoxy,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkylcarbon- yl, aryl, aryl-C.sub.1-C.sub.6-alkyl,
aryl-C.sub.1-C.sub.6-alkoxy,
aryl-C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
aryl-C.sub.1-C.sub.6-alkoxycarbonyl, cycloalkyl,
cycloalkyl-C.sub.1-C.sub- .6-alkyl,
cycloalkyl-C.sub.1-C.sub.6-alkoxy, cycloalkyl-C.sub.1-C.sub.6-al-
koxy-C.sub.1-C.sub.6-alkyl, and
cycloalkyl-C.sub.1-C.sub.6-alkoxycarbonyl. The alkyl, alkoxy,
alkoxyalkyl, alkylcarbonyl, aryl, arylalkyl, arylalkoxy,
arylalkoxyalkyl, or arylalkoxycarbonyl substituent(s) may further
be substituted with, for example, one or more halogen. The aryls or
cycloalkyls are typically single-ring groups containing from 3 to 6
ring atoms, and more typically from 5 to 6 ring atoms.
[0147] An aryl or heteroaryl optionally can be substituted with,
for example, one or more substituents independently selected from
the group consisting of halogen, --OH, --CN, --NO.sub.2, --SH,
--C(O)--OH, amino, aminocarbonyl, aminoalkyl, alkyl, alkylthio,
carboxyalkylthio, alkylcarbonyl, alkylcarbonyloxy, alkoxy,
alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkoxy, alkoxyalkylthio,
alkoxycarbonylalkylthio, carboxyalkoxy, alkoxycarbonylalkoxy,
carbocyclyl, carbocyclylalkyl, carbocyclyloxy, carbocyclylthio,
carbocyclylalkylthio, carbocyclylamino, carbocyclylalkylamino,
carbocyclylcarbonylamino, carbocyclylcarbonyl, carbocyclylalkyl,
carbonyl, carbocyclylcarbonyloxy, carbocyclyloxycarbonyl,
carbocyclylalkoxycarbonyl, carbocyclyloxyalkoxycarbocyclyl,
carbocyclylthioalkylthiocarbocyclyl,
carbocyclylthioalkoxycarbocyclyl,
carbocyclyloxyalkylthiocarbocyclyl, heterocyclyl,
heterocyclylalkyl, heterocyclyloxy, heterocyclylthio,
heterocyclylalkylthio, heterocyclylamino, heterocyclylalkylamino,
heterocyclylcarbonylamino, heterocyclylcarbonyl,
heterocyclylalkylcarbony- l, heterocyclyloxycarbonyl,
heterocyclylcarbonyloxy, heterocyclylalkoxycarbonyl,
heterocyclyloxyalkoxyheterocyclyl,
heterocyclylthioalkylthioheterocyclyl,
heterocyclylthioalkoxyheterocyclyl- , and
heterocyclyloxyalkylthioheterocyclyl. More typically, an aryl or
heteroaryl may, for example, optionally be substituted with one or
more substituents independently selected from the group consisting
of halogen, --OH, --CN, --NO.sub.2, --SH, --C(O)--OH, amino,
aminocarbonyl, amino-C.sub.1-C.sub.6-alkyl, C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkylthio, carboxy-C.sub.1-C.sub.6-alkylthio,
C.sub.1-C.sub.6-alkylcarbonyl, C.sub.1-C.sub.6-alkylcarbonyloxy,
C.sub.1-C.sub.6-alkoxy,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkyl,
C.sub.1-C.sub.6-alkoxycarbonyl,
C.sub.1-C.sub.6-alkoxycarbonyl-C.sub.1-C.- sub.6-alkoxy,
C.sub.1-C.sub.6-alkoxy-C.sub.1-C.sub.6-alkylthio,
C.sub.1-C.sub.6-alkoxycarbonyl-C.sub.1-C.sub.6-alkylthio,
carboxy-C.sub.1-C.sub.6-alkoxy,
C.sub.1-C.sub.6-alkoxycarbonyl-C.sub.1-C.- sub.6-alkoxy, aryl,
aryl-C.sub.1-C.sub.6-alkyl, aryloxy, arylthio,
aryl-C.sub.1-C.sub.6-alkylthio, arylamino,
aryl-C.sub.1-C.sub.6-alkylamin- o, arylcarbonylamino, arylcarbonyl,
aryl-C.sub.1-C.sub.6-alkylcarbonyl, arylcarbonyloxy,
aryloxycarbonyl, aryl-C.sub.1-C.sub.6-alkoxycarbonyl,
aryloxy-C.sub.1-C.sub.6-alkoxyaryl,
arylthio-C.sub.1-C.sub.6-alkylthioary- l,
arylthio-C.sub.1-C.sub.6-alkoxyaryl,
aryloxy-C.sub.1-C.sub.6-alkylthioa- ryl, cycloalkyl,
cycloalkyl-C.sub.1-C.sub.6-alkyl, cycloalkyloxy, cycloalkylthio,
cycloalkyl-C.sub.1-C.sub.6-alkylthio, cycloalkylamino,
cycloalkyl-C.sub.1-C.sub.6-alkylamino, cycloalkylcarbonylamino,
cycloalkylcarbonyl, cycloalkyl-C.sub.1-C.sub.6-alkylcarbonyl,
cycloalkylcarbonyloxy, cycloalkyloxycarbonyl,
cycloalkyl-C.sub.1-C.sub.6-- alkoxycarbonyl, heteroaryl,
heteroaryl-C.sub.1-C.sub.6-alkyl, heteroaryloxy, heteroarylthio,
heteroaryl-C.sub.1-C.sub.6-alkylthio, heteroarylamino,
heteroaryl-C.sub.1-C.sub.6-alkylamino, heteroarylcarbonylamino,
heteroarylcarbonyl, heteroaryl-C.sub.1-C.sub.6-a- lkylcarbonyl,
heteroaryloxycarbonyl, heteroarylcarbonyloxy, and
heteroaryl-C.sub.1-C.sub.6-alkoxycarbonyl. Here, one or more
hydrogens bound to a carbon in any such group may, for example,
optionally be replaced with halogen. In addition, the cycloalkyl,
aryl, and heteroaryl are typically single-ring groups containing 3
to 6 ring atoms, and more typically 5 or 6 ring atoms.
[0148] In some embodiments, an aryl or heteroaryl optionally is
substituted with one or more substituents independently selected
from the group consisting of cyano, perfluoroalkyl,
trifluoromethoxy, trifluoromethylthio, haloalkyl,
trifluoromethylalkyl, aralkoxycarbonyl, aryloxycarbonyl, hydroxy,
halo, alkyl, alkoxy, nitro, thiol, hydroxycarbonyl, aryloxy,
arylthio, aralkyl, aryl, arylcarbonylamino, heteroaryloxy,
heteroarylthio, heteroaralkyl, cycloalkyl, heterocylyloxy,
heterocylylthio, heterocylylamino, cycloalkyloxy, cycloalkylthio,
heteroaralkoxy, heteroaralkylthio, aralkoxy, aralkylthio,
aralkylamino, heterocylyl, heteroaryl, arylazo,
hydroxycarbonylalkoxy, alkoxycarbonylalkoxy, alkanoyl,
arylcarbonyl, aralkanoyl, alkanoyloxy, aralkanoyloxy, hydroxyalkyl,
hydroxyalkoxy, alkylthio, alkoxyalkylthio, alkoxycarbonyl,
aryloxyalkoxyaryl, arylthioalkylthioaryl, aryloxyalkylthioaryl,
arylthioalkoxyaryl, hydroxycarbonylalkoxy,
hydroxycarbonylalkylthio, alkoxycarbonylalkoxy,
alkoxycarbonylalkylthio, amino, aminocarbonyl, and aminoalkyl.
Here, the amino nitrogen optionally is substituted with:
[0149] (i) up two substituents that are independently selected from
the group consisting of alkyl, aryl, heteroaryl, aralkyl,
cycloalkyl, aralkoxycarbonyl, alkoxycarbonyl, arylcarbonyl,
aralkanoyl, heteroarylcarbonyl, heteroaralkanoyl, and alkanoyl;
or
[0150] (ii) two substituents such that the two substituents,
together with the amino nitrogen, form a 5- to 8-member
heterocyclyl or heteroaryl ring that:
[0151] (a) contains from zero to two additional heteroatoms that
are independently selected from the group consisting of nitrogen,
oxygen, and sulfur;
[0152] (b) optionally is substituted with up to two substituents
independently selected from the group consisting of aryl, alkyl,
heteroaryl, aralkyl, heteroaralkyl, hydroxy, alkoxy, alkanoyl,
cycloalkyl, heterocylylalkyl, alkoxycarbonyl, hydroxyalkyl,
trifluoromethyl, benzofused heterocylylalkyl, hydroxyalkoxyalkyl,
aralkoxycarbonyl, hydroxycarbonyl, aryloxycarbonyl, benzofused
heterocylylalkoxy, benzofused cycloalkylcarbonyl,
heterocyclylalkylcarbon- yl, and cycloalkylcarbonyl.
[0153] The aminocarbonyl nitrogen is:
[0154] (i) unsubstituted;
[0155] (ii) the reacted amine of an amino acid;
[0156] (iii) substituted with one or two substituents independently
selected from the group consisting of alkyl, hydroxyalkyl,
hydroxyheteroaralkyl, cycloalkyl, aralkyl, trifluoromethylalkyl,
heterocylylalkyl, benzofused heterocylylalkyl, benzofused
cycloalkyl, and N,N-dialkylsubstituted alkylaminoalkyl; or
[0157] (iv) substituted with two substituents such that the two
substituents, together with the aminocarbonyl nitrogen, form a 5-
to 8-member heterocyclyl or heteroaryl ring that optionally is
substituted with up to two substituents independently selected from
the group consisting of alkyl, alkoxycarbonyl, nitro,
heterocylylalkyl, hydroxy, hydroxycarbonyl, aryl, aralkyl,
heteroaralkyl, and amino, wherein the amino nitrogen optionally is
substituted with:
[0158] (a) two substituents independently selected from the group
consisting of alkyl, aryl, and heteroaryl; or
[0159] (b) two substituents such that the two substituents,
together with the amino nitrogen, form a 5- to 8-member
heterocyclyl or heteroaryl ring.
[0160] The aminoalkyl nitrogen optionally is substituted with:
[0161] (i) up to two substituents independently selected from the
group consisting of alkyl, aryl, aralkyl, cycloalkyl,
aralkoxycarbonyl, alkoxycarbonyl, and alkanoyl; or
[0162] (ii) two substituents such that the two substituents,
together with the aminoalkyl nitrogen, form a 5- to 8-member
heterocyclyl or heteroaryl ring.
[0163] A prefix attached to a multi-component group only applies to
the first component. To illustrate, the term "alkylcycloalkyl"
contains two components: alkyl and cycloalkyl. Thus, the
C.sub.1-C.sub.6- prefix on C.sub.1-C.sub.6-alkylcycloalkyl means
that the alkyl component of the alkylcycloalkyl contains from 1 to
6 carbon atoms; the C.sub.1-C.sub.6-prefix does not describe the
cycloalkyl component. To illustrate further, the prefix "halo" on
haloalkoxyalkyl indicates that only the alkoxy component of the
alkoxyalkyl group is substituted with one or more halogen radicals.
If halogen substitution may alternatively or additionally occur on
the alkyl component, the group would instead be described as
"halogen-substituted alkoxyalkyl" rather than "haloalkoxyalkyl."
And finally, if the halogen substitution may only occur on the
alkyl component, the group would instead be described as
"alkoxyhaloalkyl."
[0164] If substituents are described as being "independently
selected" from a group, each substituent is selected independent of
the other. Each substituent therefore may be identical to or
different from the other substituent(s).
[0165] When words are used to describe a substituent, the
rightmost-described component of the substituent is the component
that is bound at the location of the replaced hydrogen. To
illustrate, benzene substituted with methoxyethyl has the following
structure: 27
[0166] As can be seen, the ethyl is bound to the benzene, and the
methoxy is the component of the substituent that is the component
furthest from the benzene. As further illustration, benzene
substituted with cyclohexanylthiobutoxy has the following
structure: 28
[0167] When words are used to describe a linking element between
two other elements of a depicted chemical structure, the
rightmost-described component of the substituent is the component
that is bound to the left element in the depicted structure. To
illustrate, if the chemical structure is X-L-Y and L is described
as methylcyclohexanylethyl, the chemical would be
X-ethyl-cyclohexanyl-methyl-Y.
[0168] When a chemical formula is used to describe a substituent,
the dash on the left side of the formula indicates the portion of
the substituent that is bound at the location of the replaced
hydrogen. To illustrate, benzene substituted with --C(O)--OH has
the following structure: 29
[0169] When a chemical formula is used to describe a linking
element between two other elements of a depicted chemical
structure, the leftmost dash of the substituent indicates the
portion of the substituent that is bound to the left element in the
depicted structure. The rightmost dash, on the other hand,
indicates the portion of the substituent that is bound to the right
element in the depicted structure. To illustrate, if the depicted
chemical structure is X-L-Y and L is described as --C(O)--N(H)--,
the chemical would be: 30
[0170] The term "pharmaceutically acceptable" is used adjectivally
in this patent to mean that the modified noun is appropriate for
use as a pharmaceutical product or as a part of a pharmaceutical
product.
[0171] With reference to the use of the words "comprise" or
"comprises" or "comprising" in this patent (including the claims),
Applicants note that unless the context requires otherwise, those
words are used on the basis and clear understanding that they are
to be interpreted inclusively, rather than exclusively, and that
Applicants intend each of those words to be so interpreted in
construing this patent, including the claims below.
EXAMPLES
[0172] The following examples are merely illustrative, and not
intended to be limiting to the remainder of this disclosure in any
way.
[0173] Abbreviations are often used for reagents and solvents in
the specific examples that follow. Those abbreviations include the
following:
[0174] BOC=t-butoxycarbonyl
[0175] DEAD=diethyl azodicarboxylate
[0176] DMF=dimethylfonmamide
[0177] DMPU=1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone
[0178] EtOAc=ethyl acetate
[0179] EDC=1-ethyl-3-[3-(dimethylamino)-propyl]carbodiimide
hydrochloride
[0180] Et.sub.2O=diethyl ether
[0181] HOBT=1-hydroxybenzotriazole
[0182] MeOH=methanol
[0183] MeCl.sub.2=methylene chloride
[0184] MsCl=methanesulfonyl chloride
[0185] NMM=N-methyl morpholine
[0186] THF=tetrahydrofruan
[0187] TsCl=toluenesulfonyl chloride
[0188] THP-O-hydroxylamine=O-tetrahydropyran-hydroxylamine and
O-tetrahydro-2H-pyran-2-yl-hydroxylamine
[0189] The preparation of compounds useful in the synthesis of
compounds of the invention are provided herein below in Preparative
Examples I through XI.
Preparative Example I
Preparation of 1,1-dimethylethyl ester
4-[(hydroxyamino)-carbonyl]-4-[(phe-
noxyphenyl)-sulfonyl]-1-piperidinecarboxylic acid
[0190] 31
[0191] Part A: A solution of 4-(phenoxy)benzenethiol (2.03 g, 10.0
mmol) in DMSO (DMSO; 20 mL) was heated to 65.degree. C. for 5 hr.
The solution remained at ambient temperature for 18 hr. The
solution was extracted with ethyl acetate and the combined organic
layers were washed with H.sub.2O and saturated NaCl and dried over
magnesium sulfate. Concentration in vacuo provided the disulfide as
a yellow oil (2.3 g, quantitative yield).
[0192] Part B: To a solution of ethyl isonipecotate (15.7 g, 0.1
mol) in THF (100 mL) was added a solution of di-tert-butyl
dicarbonate (21.8 g, 0.1 mol) in THF (5 mL) drop-wise over 20 min.
The solution was stirred overnight (about 18.degree. C.) at ambient
temperature and concentrated in vacuo to yield a light oil. The oil
was filtered through silica gel (7:3 ethyl acetate/hexanes) and
concentrated in vacuo to give the BOC-piperidine compound (26.2 g,
quantitative yield) as a clear, colorless oil.
[0193] Part C: To a solution of diisopropylamine (2.8 mL, 20 mmoL)
in THF (30 mL), cooled to -78.degree. C., was added n-butyl lithium
(12.5 mL, 20 mmol) drop-wise. After 15 min, the BOC-piperidine
compound of part B (2.6 g, 10 mmol) in THF (10 mL) was added
drop-wise. After 1.5 hr, the solution was cooled to -60.degree. C.
and the disulfide of part A (2.0 g, 10 mmol) in THF (7 mL). The
solution was stirred at ambient temperature for 2 hr. The solution
was diluted with H.sub.2O and extracted with ethyl acetate. The
organic layer was washed with H.sub.2O and saturated NaCl and dried
over magnesium sulfate. Chromatography (on silica, ethyl
acetate/hexane) provided the sulfide as an oil (1.8 g, 40%).
[0194] Part D: To a solution of the sulfide of part C (1.8 g, 3.95
mmol) in dichloromethane (75 mL) cooled to 0.degree. C., was added
m-chloroperbenzoic acid (1.7 g, 7.9 mmol). The solution was stirred
for 1.5 hr followed by dilution with H.sub.2O and extraction with
dichloromethane. The organic layer was washed with 10 percent
Na.sub.2SO.sub.4, H.sub.2O, and saturated NaCl and dried over
magnesium sulfate. Chromatography (on silica, ethyl acetate/hexane)
provided the sulfone as a solid (1.15 g, 59%).
[0195] Part E: To a solution of the sulfone of part D (800 mg, 1.63
mmol) in THF (9 mL) and ethanol (9 mL) was added NaOH (654 mg, 16.3
mmol) in H.sub.2O (3 mL). The solution was heated at 65.degree. C.
for 18 hr. The solution was concentrated in vacuo and the residue
was dissolved in H.sub.2O. Following acidification with 2N HCl to
pH 4, the solution was extracted with ethyl acetate and the organic
layer was washed with saturated NaCl and dried over magnesium
sulfate. Concentration in vacuo provided the acid as a white foam
(790 mg, quantitative yield). Analytical calculated for
C.sub.23H.sub.27NO.sub.7S: C, 59.86; H, 5.90; N, 3.04; S, 6.95.
Found: C, 59.49; H, 6.37; N, 2.81; S, 6.59.
[0196] Part F: To a solution of the acid of part G (730 mg, 1.58
mmol) in DMF (9 mL) was added HOBT (256 mg, 1.90 mmol) followed by
EDC (424 mg, 2.21 mmol), 4-methylmorpholine (0.521 mL, 4.7 mmol)
and 50 percent aqueous hydroxylamine (1.04 mL, 15.8 mmol). The
solution was stirred for 20 hr and additional
N-hydroxybenzotriazole.H.sub.2O (256 mg), EDC (424 mg) and 50
percent aqueous hydroxylamine (1.04 mL) were added. After an
additional 24 hr of stirring, the solution was diluted with
H.sub.2O and extracted with ethyl acetate and the organic layer was
washed with saturated NaCl and dried over magnesium sulfate.
Reverse phase chromatography (on silica, acetonitrile/H.sub.2O)
provided the title compound as a white solid (460 mg, 61%). HPLC
purity: >99%. Analytical calculated for
C.sub.23H.sub.28N.sub.2O.sub.7S: C, 57.97; H, 5.92; N, 5.88; S,
6.73. Found: C, 57.95; H, 6.02; N, 5.81; S, 6.85.
Preparative Example II
Preparation of
N-hydroxy-4-[[4-(phenylthio)phenyl]sulfonyl]-1-(2-propynyl)-
-4-piperidinecarboxamide, monohydrochloride
[0197] 32
[0198] Part A: To a solution of ethyl isonipecotate (15.7 g, 0.1
mol) in THF (100 mL) was added a solution of di-tert-butyl
dicarbonate (21.8 g, 0.1 mol) in THF (5 mL) drop-wise over 20 min.
The solution was stirred overnight (about 18 hr) at ambient
temperature and concentrated in vacuo to yield a light oil. The oil
was filtered through silica gel (ethyl acetate/hexanes) and
concentrated in vacuo to give the BOC-piperidine compound as a
clear, colorless oil (26.2 g, quantitative yield).
[0199] Part B: A solution of 4-fluorothiophenol (50.29 g, 390 mmol)
in DMSO (500 mL) was heated to 65.degree. C. for 6 hr. The reaction
was quenched into wet ice and the resulting solid was collected by
vacuum filtration to provide the disulfide as a white solid (34.4
g, 68.9%).
[0200] Part C: To a solution of the BOC-piperdine compound of part
A (16 g, 62 mmol) in THF (300 mL) cooled to minus 50.degree. C. was
added lithium diisopropylamide (41.33 mL, 74 mmol) and the solution
was stirred for 1.5 hr at 0.degree. C. To this solution was added
the disulfide of part B (15.77 g, 62 mmol), and the resulting
solution was stirred at ambient temperature for 20 hr. The reaction
was quenched with the addition of H.sub.2O and the solution was
concentrated in vacuo. The aqueous residue was extracted with ethyl
acetate and the organic layer was washed with 0.5N KOH, H.sub.2O,
and saturated NaCl. Chromatography (on silica, hexane/ethyl
acetate) provided the sulfide as an oil (18.0 g, 75%).
[0201] Part D: To a solution of the sulfide of part C (16.5 g, 43
mmol) in dichloromethane (500 mL) cooled to 0.degree. C. was added
3-chloroperbenzoic acid (18.0 g, 86 mmol) and the solution was
stirred for 20 hr. The solution was diluted with H.sub.2O and
extracted with dichloromethane. The organic layer was washed with
10 percent Na.sub.2SO.sub.3, H.sub.2O, and saturated NaCl and dried
over magnesium sulfate. Chromatography (on silica, ethyl
acetate/hexane) provided the sulfone as a solid (10.7 g, 60%).
[0202] Part E: Into a solution of the sulfone of part D (10 g, 24.0
mmol) in ethyl acetate (250 mL) was bubbled HCl gas for 10 min
followed by stirring at ambient temperature for 4 hr. Concentration
in vacuo provided the amine hydrochloride salt as a white solid
(7.27 g, 86%).
[0203] Part F: To a solution of the amine hydrochloride salt of
part E (5.98 g, 17.0 mmol) in DMF (120 mL) was added potassium
carbonate (4.7 g, 34.0 mmol) followed by propargyl bromide (2.02 g,
17.0 mmol) and the solution was stirred for 4 hr at ambient
temperature. The solution was partitioned between ethyl acetate and
H.sub.2O, and the organic layer was washed with H.sub.2O and
saturated NaCl and dried over magnesium sulfate. Chromatography (on
silica, ethyl acetate/hexane) provided the propargyl amine as a
yellow oil (5.2 g, 86%).
[0204] Part G: To a solution of the propargyl amine of part F in
DMF (15 mL) was added thiophenol (0.80 mL, 7.78 mmol) and
CsCO.sub.3 (2.79 g, 8.56 mmol) and the solution was heated to
70.degree. C. for 6 hr. The solution was partitioned between ethyl
ether and H.sub.2O. The organic layer was washed with H.sub.2O and
saturated NaCl, and dried over magnesium sulfate. Chromatography
(on silica, ethyl acetate/hexane) provided the S-phenoxyphenyl
compound as an oil (1.95 g, 56%).
[0205] Part H: To a solution of the S-phenoxyphenyl of part G (1.81
g, 4.06 mmol) in ethanol (21 mL) and H.sub.2O (3.5 mL) was added
KOH (1.37 g, 24.5 mmol) and the solution was heated to 105.degree.
C. for 4.5 hr. The solution was acidified to a pH value of 1 with
concentrated HCl solution and then concentrated to provide the acid
as a yellow residue that was used without additional purification
(1.82 g).
[0206] Part I: To a solution of the acid of part H (1.82 g, 4.06
mmol) in acetonitrile (20 mL) was added
O-tetrahydro-2H-pyran-2-yl-hydroxylamine (723 mg, 6.17 mmol) and
triethylamine (0.67 mL, 4.86 mmol). To this stirring solution was
added EDC (1.18 g, 6.17 mmol) and the solution was stirred for 18
hr. The solution was partitioned between H.sub.2O and ethyl
acetate. The organic layer was washed with H.sub.2O, saturated
NaHCO.sub.3 and saturated NaCl and dried over magnesium sulfate.
Chromatography (on silica, ethyl acetate/hexane) provided the
protected hydroxamate as a white solid (1.32 g, 63%).
[0207] Part J: To a solution of the protected hydroxamate of part 1
(9.65 g, 18.7 mmol) in methanol (148 mL) cooled to 0.degree. C. was
added acetyl chloride (4.0 mL, 56.2 mmol), and the solution was
stirred for 45 min at ambient temperature. Concentration in vacuo
followed by trituration with ethyl ether provided the title
compound as a white solid (8.10 g, 94%). MS(CI) MH.sup.+ calculated
for C.sub.21H.sub.22N.sub.2O.su- b.4S.sub.2: 431, found 431.
Preparative Example III
Preparation of
N-hydroxy-4-[(4-phenoxyphenyl)sulfonyl]-1-(2-propynyl)-4-pi-
peridinecarboxamide, monohydrochloride
[0208] 33
[0209] Part A: A solution of 4-(phenoxy)benzenethiol (2.03 g, 10.0
mmol) in DMSO (20 mL) was heated to 65.degree. C. for 5 hr. The
solution remained at ambient temperature for 18 hr. The solution
was extracted with ethyl acetate and the combined organic layers
were washed with H.sub.2O and saturated NaCl, and dried over
magnesium sulfate. Concentration in vacuo provided the disulfide as
a yellow oil (2.3 g, quantitative yield).
[0210] Part B: To a solution of ethyl isonipecotate (15.7 g, 0.1
mol) in THF (100 mL) was added a solution of di-tert-butyl
dicarbonate (21.8 g, 0.1 mol) in THF (5 mL) dropwise over 20 min.
The solution was stirred overnight at ambient temperature and
concentrated in vacuo to yield a light oil. The oil was filtered
through silica gel (ethyl acetate/hexane) and concentrated in vacuo
to give the BOC-piperidine compound as a clear, colorless oil (26.2
g, quantitative yield).
[0211] Part C: To a solution of diisopropylamine (2.8 mL, 20 mmoL)
in THF (30 mL), cooled to -78.degree. C., was added n-butyl lithium
(12.5 mL, 20 mmol) dropwise. After 15 min, the BOC-piperidine
compound of Part B (2.6 g, 10 mmol) in THF (10 mL) was added
dropwise. After 1.5 hr, the solution was cooled to -60.degree. C.
and the disulfide of Part A (2.0 g, 10 mmol) in THF (7 mL) was
added. The solution was stirred at ambient temperature for 2 hr.
The solution was diluted with H.sub.2O and extracted with ethyl
acetate. The organic layer was washed with H.sub.2O and saturated
NaCl and dried over magnesium sulfate. Chromatography (on silica,
ethyl acetate/hexane) provided the sulfide as an oil (1.8 g,
40%).
[0212] Part D: To a solution of the sulfide of Part C (1.8 g, 3.95
mmol) in dichloromethane (75 mL) cooled to 0.degree. C., was added
m-chloroperbenzoic acid (1.7 g, 7.9 mmol). The solution was stirred
for 1.5 hr followed by dilution with H.sub.2O and extraction with
dichloromethane. The organic layer was washed with 10 percent
Na.sub.2SO.sub.4, H.sub.2O, and saturated NaCl and dried over
magnesium sulfate. Chromatography (on silica, ethyl acetate/hexane)
provided the sulfone as a solid (1.15 g, 59%).
[0213] Part E: Into a solution of the sulfone of Part D (3.56 g,
7.0 mmol) in ethyl acetate (100 mL) cooled to 0.degree. C. was
bubbled HCl gas for 5 min. Concentration in vacuo followed by
trituration with ethyl ether provided the amine hydrochloride salt
as a white solid (3.5 g, quantitative yield). MS(CI) MH.sup.+
calculated for C.sub.20H.sub.23NO.sub.5S: 390, found 390.
[0214] Part F: To a solution of the amine hydrochloride salt of
part E (2.6 g, 6 mmol) and K.sub.2CO.sub.3 (1.66 g, 12 mmol) in DMF
(50 mL) was added propargyl bromide (892 mg, 6 mmol) and the
solution was stirred at ambient temperature for 4 hr. The solution
was diluted with H.sub.2O and extracted with ethyl acetate. The
combined organic layers were washed with saturated NaCl and dried
over magnesium sulfate. Chromatography (on silica, ethyl
acetate/hexane) provided the propargyl amine as a white solid (2.15
g, 82%).
[0215] Part G: To a solution of the propargyl amine of part F (2.15
g, 5 mmol) in THF (30 mL) and ethanol (30 mL) was added NaOH (2.0
g, 50 mmol) and the solution was heated at 65.degree. C. for 48 hr.
The solution was concentrated in vacuo and the aqueous residue was
acidified to a pH value of 5. Vacuum filtration of the resulting
precipitate provided the acid as a white solid (2.04 g,
quantitative yield).
[0216] Part H: To a solution of the acid of part G (559 mg, 1.4
mmol) in dichloromethane (5 mL) was added triethylamine (0.585 mL,
4.2 mmol) and 50 percent aqueous hydroxylamine (0.925 mL, 14.0
mmol) followed by bromotris(pyrrolidino)phosphonium
hexafluourphosphate (PyBroP.RTM.; 718 mg, 1.54 mmol). The solution
was stirred at ambient temperature for 4 hr. The solution was
diluted with H.sub.2O and extracted with dichloromethane. The
organic layer was washed with saturated NaCl and dried over
magnesium sulfate. Reverse phase chromatography (on silica,
acetonitrile/H.sub.2O) provided the hydroxamate as a white solid
(140 mg, 25%). Analytical calculation for
C.sub.21H.sub.22N.sub.2O.sub.5S: C, 60.85; H, 5.37; N, 6.76; S,
7.74. Found: C, 60.47; H, 5.35; N, 6.61; S, 7.46.
[0217] Part I: To a solution of the hydroxamate of part H (121 mg,
0.292 mmol) in methanol (2 mL) cooled to 0.degree. C. was added
acetyl chloride (0.228 mL, 0.321 mmol) in methanol (1 mL). After
stirring at ambient temperature for 30 min, the solution was
concentrated under a stream of N.sub.2. Trituration with ethyl
ether provided the title compound as a white solid (107 mg, 81%).
Analytical calculation for
C.sub.21H.sub.22N.sub.2O.sub.5S.HCl.0.3H.sub.2O: C, 55.27; H, 5.21;
N, 6.14. Found: C, 54.90; H, 5.37; N, 6.07.
Preparative Example IV
Preparation of
4-[(4-fluorophenyl)sulfonyl]tetrahydro-N-[(tetrahydro-2H-py-
ran-2-yl)oxy]-2H-pyran-4-carboxamide
[0218] 34
[0219] Part A: In dry equipment under nitrogen, sodium metal (8.97
g, 0.39 mol) was added to methanol (1000 mL) at 2.degree. C. The
reaction was stirred at ambient temperature for 45 min, at which
time the sodium had dissolved. The solution was chilled to
5.degree. C. and p-fluorothiophenol (41.55 mL, 0.39 mmol) was
added, followed by methyl 2-chloroacetate (34.2 mL, 0.39 mol). The
reaction was stirred at ambient temperature for 4 hr, filtered, and
concentrated in vacuo to give the sulfide as a clear colorless oil
(75.85 g, 97%).
[0220] Part B: To a solution of the sulfide from part A (75.85 g,
0.38 mol) in methanol (1000 mL) were added water (100 mL) and
Oxone.RTM. (720 g, 1.17 mol) at 20.degree. C. An exotherm to
67.degree. C. was noted. After 2 hr, the reaction was filtered and
the cake was washed well with methanol. The filtrate was
concentrated in vacuo. The residue was taken up in ethyl acetate
and washed with brine, dried over MgSO.sub.4, filtered, and
concentrated in vacuo to give the sulfone as a crystalline solid
(82.74 g, 94%).
[0221] Part C: To a solution of the sulfone from part B (28.5 g,
0.123 mol) in N,N-dimethylacetamide (200 mL) were added potassium
carbonate (37.3 g, 0.27 mol), bis-(2-bromoethyl)ether (19.3 mL,
0.147 mol), 4-dimethylaminopyridine (0.75 g, 6 mmol), and
tetrabutylammonium bromide (1.98 g, 6 mmol). The reaction was
stirred overnight (about 18 hr) at ambient temperature. The
reaction was slowly poured into 1N HCl (300 mL), the resultant
solid filtered and the cake washed well with hexanes. The solid was
recrystallized from ethyl acetate/hexanes to give the pyran
compound as a beige solid (28.74 g, 77%). MS (ES+) MH+ calculated
for C.sub.13H.sub.15O.sub.5S.sub.1F.sub.1: 303, found 303.
[0222] Part D: In dry equipment under nitrogen, the pyran compound
from part C (8.0 g, 26.5 mmol) was dissolved in dry tetrahydrofuran
(250 mL) and a solution of potassium trimethylsilonate (10.2 g,
79.5 mmol) in dry tetrahydrofuran (15 mL) was added at ambient
temperature. After 90 min, water (100 mL) was added and the
solution concentrated in vacuo. The residue was taken up in water
and extracted with ethyl acetate to remove unreacted starting
material. The aqueous solution was treated with 6N HCl until
pH.dbd.I. The slurry was extracted with ethyl acetate and the
combined extracts washed with water, dried over Na.sub.2SO.sub.4,
filtered, and concentrated in vacuo. The residue was heated in
diethyl ether, the solid filtered and dried to give the carboxylic
acid as a crystalline solid (5.78 g, 76%). HRMS (ES-) M-H
calculated for C.sub.12H.sub.13O.sub.5S.sub.1F.sub.1: 287.04, found
287.04.
[0223] Part E: In dry equipment under nitrogen, the carboxylic acid
from part D (9.1 g, 31.6 mmol) was dissolved in dry
N,N-dimethylformamide (70 mL) and the remaining reagents were added
to the solution in the following order: N-hydroxybenzotriazole
hydrate (5.1 g, 37.9 mmol), N-methylmorpholine (10.4 mL, 94.8
mmol), O-tetrahydro-2H-pyran-2-yl-hydro- xylamine (11.5 g, 98
mmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiim- ide
hydrochloride (8.48 g, 44.2 mmol). After 3 hr at ambient
temperature, the reaction was concentrated in vacuo. The residue
was taken up in ethyl acetate, washed with water, 5% KHSO.sub.4,
saturated NaHCO.sub.3, brine, dried over Na.sub.2SO.sub.4,
filtered, and concentrated in vacuo. Chromatography (on silica,
ethyl acetate/hexanes) provided the title compound as a crystalline
solid (9.7 g, 80%). HRMS (ES+) MH+ calculated for
C.sub.17H.sub.22NO.sub.6S.sub.1F.sub.1: 388.12, found 388.12.
Preparative Example V
Preparation of
tetrahydro-N-hydroxy-4-[[4-[4-trifluoromethoxy)-phenoxy)phe-
nyl]sulfonyl]-2H-pyran-4-carboxamide
[0224] 35
[0225] Part A: To a solution of the title compound of Preparative
Example TV (3.1 g, 8 mmol) in N,N-dimethylacetamide (20 mL) were
added cesium carbonate (8.8 g, 27 mmol) and
p-(trifluoromethoxy)phenol (2.1 mL, 16 mmol). The slurry was
stirred at 95.degree. C. for 19 hr. The reaction was concentrated
in vacuo. The residue was taken up in ethyl acetate, washed with
brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated in
vacuo. Chromatography (on silica, ethyl acetate/hexanes) provided
the substituted THP-protected hydroxamate as a white foam (4.2 g,
96%). HRMS (ES+) MH+ calculated for
C.sub.24H.sub.26N.sub.1O.sub.8S.su- b.1F.sub.3: 546.14, found
546.14.
[0226] Part B: To a slurry of the THP-protected hydroxamate from
part A (4.0 g, 7.3 mmol) in dioxane (20 mL) were added a 4N HCl
dioxane solution (20 mL) and methanol (20 mL). After 15 min, at
ambient temperature, the reaction was diluted with ethyl acetate
and washed with water, dried over Na.sub.2SO.sub.4, filtered, and
concentrated in vacuo. The product was recrystallized
(acetone/hexanes) to give the title compound as a white solid (2.2
g, 65%). HRMS (ES+) M+NH.sub.4.sup.+ calculated for
C.sub.19H.sub.18N.sub.1O.sub.7S.sub.1F.sub.3: 479.11, found
479.11.
Preparative Example VI
Preparation of
1-cyclopropyl-N-hydroxy-4-[[4-(2-phenoxy-ethoxy)phenyl]sulf-
onyl]-4-piperidine carboxamide, monohydrochloride
[0227] 36
[0228] Part A: To a solution of the product of Preparative Example
II, part E, (14.36 g, 40 mmol) in methanol (50 mL) was added acetic
acid (24.5 g, 400 mmol), a portion (about 2 g) of 4-Angstrom
molecular sieves, (1-ethoxycyclopropyl)-oxytrimethyl silane (25.8
mL, 148 mmol) and sodium cyanoborohydride (7.05 g, 112 mmol). The
solution was heated at reflux for 8 hr. The precipitated solids
were removed by filtration and the filtrate was concentrated in
vacuo. The residue was diluted with H.sub.2O (400 mL) and extracted
with ethyl acetate. The organic layer was washed with saturated
NaCl and dried over MgSO.sub.4, filtered and concentrated in vacuo.
The solid was filtered, washed with H.sub.2O/diethyl ether to give
the desired cyclopropyl amine {ethyl
4-[(4-fluorophenyl-sulfonyl)]-1-
-cyclopropyl-4-piperidinecarboxylate} as a white solid (11.83 g,
81.5%). MS MH.sup.+ calculated for C.sub.17H.sub.22NO.sub.4SF: 356,
found: 356.
[0229] Part B: A solution of the cyclopropyl amine of Part A (2.0
g, 5.6 mmol), ethylene glycol phenyl ether (2.8 mL, 23 mmol), and
cesium carbonate (7.3 g, 23 mmol) in DMAC (10 mL) was heat at
125-135.degree. C. for 18 hr under an atmosphere of nitrogen. The
mixture was concentrated in vacuo, diluted with water, and
extracted with ethyl acetate. The combined ethyl acetate layers
were washed with water and brine, dried over magnesium sulfate,
concentrated in vacuo, dissolved in diethyl ether, precipitated as
the hydrochloride salt, and dried at 40.degree. C. in a vacuum
oven. The solid was dissolved into a mixture of water,
acetonitrile, and ethanol and then the pH was adjusted to 12 with
1N NaOH solution. The mixture was concentrated in vacuo to remove
ethanol and acetonitrile. The solid was isolated by filtration,
washed with water, and dried at 50.degree. C. in a vacuum oven to
afford the ether as a white solid (1.8 g, 68%): MS+ calcd. for
C.sub.25H.sub.31NO.sub.6S 474, found 474. Anal. calcd. for
C.sub.25H.sub.31NO.sub.6S: C, 63.40; H, 6.60; N, 2.96; S, 6.77.
Found: C, 63.35; H, 6.59; N, 2.99; S, 6.61.
[0230] Part C: A mixture of the ether of part B (1.8 g, 3.7 mmol)
and a 50% NaOH aqueous solution (3.0 g, 37 mmol) in THF (32 mL),
EtOH (32 mL), and H.sub.2O (16 mL) was heated at 60.degree. C.
under a N.sub.2 atmosphere for 24 hr. The material was concentrated
in vacuo and triturated with diethyl ether to give a solid. The tan
solid was dissolved into a mixture of water, ethanol, and THF,
precipitated by adjusting the pH to 3 with concentrated
hydrochloric acid, concentrated in vacuo, triturated with water,
and dried at 50.degree. C. in a vacuum oven to give a crude white
solid acid (2.3 g).
[0231] A mixture of the crude white solid acid (2.3 g),
N-hydroxybenzotriazole (1.9 g, 14 mmol), 4-methylmorpholine (1.6
mL, 14 mmol), O-tetrahydro-2H-pyran-2-yl-hydroxylamine (1.1 g, 9.4
mmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride (2.7 g, 14 mmol) in DMF (90 mL) was stirred at
ambient temperature under a nitrogen atmosphere for 2 days. The
mixture was concentrated in vacuo, diluted with water, and
extracted with ethyl acetate. The organic layer was washed with 1N
NaOH solution, water, and brine, dried over magnesium sulfate,
concentrated in vacuo, and purification by flash chromatography
(20:80 to 40:60 ethyl acetate/toluene) to afford the protected
hydroxamate as a white solid: (0.43 g, 21%): MS MH+ calcd. for
C.sub.28H.sub.36N.sub.2O.sub.7S 545, found 545. Anal. calcd. for
C.sub.28H.sub.36N.sub.2O.sub.7S: C, 61.74; H, 6.66; N, 5.14; S,
5.89. Found: C, 61.72; H, 6.75; N, 5.06; S, 5.91.
[0232] Additional compound was isolated by acidifying the aqueous
layer to pH of 3, collecting the solid by filtration, and drying to
give a white solid (0.80 g).
[0233] Part D: To an ambient temperature solution of acetyl
chloride (0.31 mL, 4.4 mmol) in methanol (11 mL) under a nitrogen
atmosphere was added the protected hydroxamate of part C (0.80 g,
1.5 mmol). After stirring for 2.5 hr, the precipitate was collected
by filtration, washed with diethyl ether, and dried at 45.degree.
C. in a vacuum oven to afford the title compound as a white solid
(0.58 g, 79%): MS MH+ calcd. for C.sub.23H.sub.28N.sub.2O.sub.6S
461, found 461. Anal. calcd. for
C.sub.23H.sub.28N.sub.2O.sub.6S.1.5HCl: C, 53.62; H, 5.77; N, 5.44;
S, 6.22. Found: C, 53.47; H, 5.79; N, 5.41; S, 6.16.
Preparative Example VII
Preparation of
N-hydroxy-1-(2-methoxyethyl)-4-[[4-[4-(trifluoro-methoxy)ph-
enoxy]phenyl]sulfonyl]-4-piperidinecarboxamide,
monohydrochloride
[0234] 37
[0235] Part A: To a solution of the product of Preparative Example
II, Part D (30 g, 161 mmol) in dichloromethane (50 mL) cooled to
0.degree. C. was added trifluroacetic acid (25 mL) and the solution
was stirred at ambient temperature for 1 hr. Concentration in vacuo
provided the amine trifluoroacetate salt as a light yellow gel. To
the solution of the trifluoroacetate salt and K.sub.2CO.sub.3 (3.6
g, 26 mmol) in N,N-dimethylformamide (50 mL) cooled to 0.degree. C.
was added 2-bromoethyl methyl ether (19 mL, 201 mmol), and solution
was stirred at ambient temperature for 36 hr. Then,
N,N-dimethylformamide was evaporated under high vacuum and the
residue was diluted with ethyl acetate. The organic layer was
washed with water and dried over MgSO.sub.4. Concentration in vacuo
provided the methoxyethyl amine as a light yellow gel (26.03 g,
86.8%).
[0236] Part B: To a solution of methoxyethyl amine (6.0 g, 16.0
mmol) of Part A and powdered K.sub.2CO.sub.3 (4.44 g, 32 mmol) in
N,N-dimethylformamide (30 mL) was added 4-(trifluoromethoxy)phenol
(5.72 g, 32 mmol) at ambient temperature and the solution was
heated to 90.degree. C. for 25 hr. The solution was concentrated
under high vacuum and the residue was dissolved in ethyl acetate.
The organic layer was washed with 1N NaOH, H.sub.2O and dried over
MgSO.sub.4. Chromatography on silica eluting with ethyl
acetate/hexane provided trifluoromethoxy phenoxyphenyl sulfone as a
light yellow gel (7.81 g, 91.5%).
[0237] Part C: To a solution of trifluoromethoxy phenoxyphenyl
sulfone of Part B (7.81 g, 14.7 mmol) in ethanol (14 mL) and
tetrahydrofuran (14 mL) was added NaOH (5.88 g, 147 mmol) in
H.sub.2O (28 mL) from an addition funnel at ambient temperature.
The solution was then heated to 60.degree. C. for 18 hr. The
solution was concentrated in vacuo and diluted with water. The
aqueous layer was extracted with ether and acidified to pH=2.
Vacuum filtration of white precipitation provided the acid as a
white solid (5.64 g, 73.3%).
[0238] Part D: To a solution of the acid of Part C (5.64 g, 10.8
mmol), N-methyl morpholine (4.8 mL, 43.1 mmol),
1-hydroxybenzotriazole (4.38 g, 32.4 mmol) and O-tetrahydropyranyl
hydroxylamine (2.5 g, 21.6 mmol) in N,N-dimethylformamide (50 mL)
was added 1-[3-(dimethylamino)propyl]-3-eth- ylcarbodiimide
hydrochloride (6.2 g, 32.4 mmol), and the solution was stirred at
ambient temperature for 24 hr. The solution was concentrated under
high vacuum and the residue was dissolved in ethyl acetate. The
organic layer was washed with saturated aqueous NaHCO.sub.3,
H.sub.2O and dried over MgSO.sub.4. Concentration in vacuo and
chromatography on silica eluting with ethyl acetate/hexane provided
the tetrahydropyranyl-protected hydroxamate as a white foam (6.65
g, quantitative yield).
[0239] Part E: To a solution of 4N HCl in dioxane (28 mL, 110 mmol)
was added a solution of the tetrahydropyranyl-protected hydroxamate
of Part D (6.65 g, 11.03 mmol) in methanol (3 mL) and dioxane (9
mL) and was stirred at ambient temperature for 3 hr. Concentration
in vacuo and trituration with diethyl ether provided the title
compound as a white solid (4.79 g, 78.2%). Analytical calculation
for C.sub.22H.sub.25N.sub.2- O.sub.7SF.sub.3.HCl.0.5H.sub.2O: C,
46.85; H, 4.83; N, 4.97; S, 5.69. Found: C, 46.73; H, 4.57; N,
4.82; S, 5.77.
Preparative Example VIII
Preparation of
N-hydroxy-1-[2-(4-morpholinyl)-ethyl]-4-[[4-[4-(trifluorome-
thyl)phenoxy]-phenyl]sulfonyl]-4-piperidinecarboxamide,
dihydrochloride
[0240] 38
[0241] Part A: To a suspension of 4-bromopiperidine hydrobromide
(107.0 g, 0.436 mol) in tetrahydrofuran (1 L) was slowly added
triethylamine (122 mL, 0.872 mol) followed by di-tert-butyl
dicarbonate (100 g, 0.458 mol), which was added in several
portions. The resulting mixture was stirred at ambient temperature
for 22 hr then filtered and concentrated in vacuo. The solids were
washed with hexanes and then collected by filtration to give the
Boc-piperidine compound as an amber oil (124 g, >100%).
[0242] Part B: To a solution of 4-fluorophenol (50.0 g, 0.390 mol)
in acetone (400 mL), degassed with N.sub.2, was added
Cs.sub.2CO.sub.3 (159 g, 0.488 mol). After degassing the resulting
mixture with N.sub.2 for 5 min, the Boc-piperidine compound of Part
A (85.9 g, 0.325 mol) was added. The resulting mixture was stirred
at ambient temperature for 18 hr and then filtered through a pad of
Celite.RTM., washing with acetone. The filtrate was concentrated in
vacuo to provide the sulfide as a tan residue (98.5 g, 97%).
[0243] Part C: To a solution of the sulfide of Part B (8.00 g, 25.7
mmol) in dichloromethane (90 mL) and methanol (15 mL) was added
monoperoxyphthalic acid magnesium salt hexahydrate (19.1 g, 38.6
mmol) in two portions. The resulting mixture was stirred at ambient
temperature for 1.5 hr and then filtered. The filtrate was washed
with saturated NaHCO.sub.3 and then with saturated NaCl. The
combined aqueous layers were extracted with dichloromethane (100
mL). The combined organic layers were dried over Na.sub.2SO.sub.4
and then concentrated in vacuo. The resulting solids were washed
with hexanes then dissolved in dichloromethane and filtered through
a pad of Celite.RTM., washing with dichloromethane. The filtrate
was concentrated in vacuo and recrystallization from ethyl acetate
provided the sulfone as a white crystalline solid (4.45 g,
50%).
[0244] Part D: To a solution of sulfone of Part C (7.00 g, 20.4
mmol) in N,N-dimethylformamide (40 mL) was added Cs.sub.2CO.sub.3
(19.9 g, 61.2 mmol) and .alpha.,.alpha.,.alpha.-trifluoro-p-cresol
(3.97 g, 24.5 mmol). The resulting mixture was heated at 80.degree.
C. for 16 hr. After cooling to ambient temperature, the reaction
mixture was concentrated in vacuo. The resulting residue was
treated with H.sub.2O and the solids were collected by filtration.
The solids were then washed with hexanes then methanol to provide
the biaryl ether as a tan solid (8.60 g, 87%).
[0245] Part E: To a solution of the biaryl ether of Part D (8.59 g,
17.7 mmol) in tetrahydrofuran (100 mL), cooled to 0.degree. C., was
slowly added lithium bis(trimethylsilyl)amide (22.0 mL, 11.0M in
tetrahydrofuran, 22.0 mmol), at such a rate that the temperature of
the reaction never exceeded 1.degree. C. The resulting mixture was
stirred at 0.degree. C. for 1 hr then a solution of methyl
chloroformate (2.05 mL, 26.6 mmol) in tetrahydrofuran (5.0 mL) was
slowly added, at such a rate that the temperature of the reaction
mixture never exceeded 4.degree. C. After the addition was
complete, the mixture was slowly permitted to warm to ambient
temperature. Saturated NH.sub.4Cl (50 mL) was added and the
tetrahydrofuran was removed in vacuo. Water (50 mL) was added to
the residue which was then extracted with ethyl acetate. The
combined organic layers were washed with saturated NaCl and dried
over Na.sub.2SO.sub.4. Recrystallization from methanol provided the
methyl ester as a pale yellow crystalline solid (7.66 g, 80%).
[0246] Part F: To a solution of the methyl ester of Part E (7.66 g,
14.1 mmol) in dioxane (30 mL) and methanol (10 mL) was added a
solution of 4N HCl in dioxane (10 mL, 40 mmol). After stirring at
ambient temperature for 2 hr, additional 4N HCl in dioxane (10 mL,
40 mmol) was added. After stirring at ambient temperature for 2.5
hr, the reaction mixture was concentrated in vacuo to provide the
amine as an off-white solid (6.80 g, >100%).
[0247] Part G: To a suspension of the amine of Part F (3.00 g, 6.25
mmol) in acetonitrile (20 mL) was added K.sub.2CO.sub.3 (3.46 g,
25.0 mmol), 4-(2-chloroethyl)morpholine hydrochloride (1.22 g, 6.56
mmol) and a catalytic amount of NaI. The resulting mixture was
heated at reflux for 22 hr. After cooling to ambient temperature,
the reaction mixture was filtered through a pad of Celite.RTM.,
washing with ethyl acetate. The filtrate was concentrated in vacuo
to provide the morpholinyl ethyl amine as a tan solid (3.45 g,
>100%).
[0248] Part H: To a solution of the morpholinyl ethyl amine of Part
G (3.45 g, 6.25 mmol) in tetrahydrofuran (60 mL) was added
potassium trimethylsilanolate (1.60 g, 12.50 mmol). After stirring
at ambient temperature for 25 hr, H.sub.2O was added. The reaction
mixture was then neutralized (pH 7) with 1N HCl. The
tetrahydrofuran was removed in vacuo and the resulting precipitate
was collected by filtration and washed with diethyl ether to
provide the amino acid as an off-white solid (2.87 g, 85%).
[0249] Part I: To a suspension of the amino acid of Part H (2.87 g,
5.29 mmol) in dichloromethane (25 mL) was added N-methylmorpholine
(1.74 mL, 15.9 mmol), O-(tetrahydropuranyl)hydroxylamine (0.682 g,
5.82 mmol) and PyBroP.RTM. (2.96 g, 6.35 mmol). After stirring at
ambient temperature for 19 hr, additional N-methylmorpholine (0.872
mL, 7.94 mmol), O-(tetrahydropuranyl) hydroxylamine (0.310 g, 2.65
mmol) and PyBroP.RTM. (1.48 g, 3.17 mmol) were added. The resulting
mixture was stirred at ambient temperature for 3 hr and then
concentrated in vacuo. The residue was partitioned between ethyl
acetate and H.sub.2O. The organic layers were washed with saturated
NaCl and dried over Na.sub.2SO.sub.4. Chromatography (on silica,
methanol/chloroform) provided the protected hydroxamate as an
off-white solid (2.62 g, 77%).
[0250] Part J: To a solution of the protected hydroxamate of Part I
(2.62 g, 4.08 mmol) in dioxane (9 mL) and methanol (3 mL) was added
a solution of 4N HCl in dioxane (10 mL, 40.0 mmol). The resulting
mixture was stirred at ambient temperature for 2 hr and then
diethyl ether (20 mL) was added. The resulting solids were
collected by filtration to give the title compound as an off-white
solid (2.31 g, 90%). MS MH.sup.+ calculated for
C.sub.25H.sub.31O.sub.6N.sub.3SF.sub.3: 558, found 558.
Preparative Example IX
Preparation of
1-cyclopropyl-N-hydroxy-4-[[4-[4-(trifluoromethoxy)phenoxy]-
-phenyl]sulfonyl]-4-piperidine-carboxamide, monohydrochloride
[0251] 39
[0252] Part A: To a solution of the product of Preparative Example
VI, Part A, (6.97 g, 19.6 mmol) in DMF (500 mL) was added
K.sub.2CO.sub.3 (3.42 g, 18.0 mmol) and 4-(triflouromethoxy)phenol
(3.7 g, 24.8 mmol). The solution was stirred at 90.degree. C. for
40 hr. The solution was diluted with H.sub.2O (600 mL) and
extracted with ethyl acetate. The organic layer was washed with
water, saturated NaCl and dried over MgSO.sub.4, filtered and
concentrated in vacuo to afford the desired diaryl ether as an oil
(8.5 g, quantitative). HRMS MH.sup.+ calculated for
C.sub.24H.sub.26NSO.sub.6F.sub.3: 514.1511. Found 514.1524.
[0253] Part B: To a solution of diaryl ether from Part A (8.4 g,
16.4 mmol) in ethanol (50 mL) and tetrahydrofuran (50 mL) was added
a solution of NaOH (6.54 g, 164 mmol) in water (20 mL) and the
solution was heated at 60.degree. C. for 18 hr. The solution was
concentrated in vacuo to remove most of organic solvents and the
aqueous residue was acidified to pH=4.0. The resulting precipitate
was filtered to give the desired filtered to give the hydrochloride
salt as a white solid (5.01 g, 63%). HRMS MH.sup.+ calculated for
C.sub.22H.sub.22NSO.sub.6F.sub.3: 486.1198, found 486.1200.
[0254] Part C: To a solution of the hydrochloride salt of Part B
(5.0 g, 10.3 mmol) in DMF (80 mL) were added 1-hydroxybenzotriazole
(1.65 g, 12.3 mmol), N-methyl morpholine (3.4 mL, 30.9 mmol) and
O-tetrahydropyranyl hydroxylamine hydrochloride (1.8 g, 15.4 mmol)
followed by 1-3-(dimethylamino)propyl]-3-ethylcarbodiimide
hydrochloride (1.60 g, 12.3 mmol). The solution was stirred at
ambient temperature for 42 hr. The solution was diluted with
H.sub.2O (400 mL) and extracted with ethyl acetate. The organic
layer was washed with saturated NaCl and dried over MgSO.sub.4,
filtered and concentrated in vacuo. Chromatography on silica gel,
eluting with 30% ethyl acetate/hexane provided the desired
tetrahydropyranyl-protected hydroxamate as a white solid (5.41 g,
89%).
[0255] Part D: To a solution of tetrahydropyranyl-protected
hydroxamate of Part C (5.4 g, 9.2 mmol) in dioxane (80 mL) and
methanol (20 mL) was added 4 N HCl/dioxane (50 mL). The reaction
was stirred at ambient temperature for 2.5 hr, the solution was
concentrated in vacuo. Trituration with diethyl ether afforded the
title compound as a white solid (4.02 g, 81%). HRMS MH.sup.+
calculated for C.sub.22H.sub.23N.sub.2- SO.sub.6F.sub.3: 501.1307,
found 501.1324.
Preparative Example X
Preparation of
1-cyclopropyl-N-hydroxy-4-[[4-[4-(trifluoromethyl)phenoxy]p-
henyl]sulfonyl]-4-piperidinecarboxamide, monohydrochloride
[0256] 40
[0257] Part A: To a solution of the product of Preparative Example
VI, Part A, (5.96 g, 15.0 mmol) in DMF (100 mL) was added
K.sub.2CO.sub.3 (12.34 g, 38.0 mmol) and
.quadrature.-trifluoromethyl phenol (3.65 g, 22.5 mmol). The
solution was stirred 90.degree. C. for 28 hr. The solution was
diluted with H.sub.2O (400 mL) and extracted with ethyl acetate.
The organic layer was washed with water, saturated NaCl and dried
over MgSO.sub.4, filtered and concentrated in vacuo to afford
desired aryl ether as an oil (7.54 g, quantitative)
[0258] Part B: To a solution of aryl ether from Part A (7.54 g,
15.0 mmol) in ethanol (40 mL) and tetrahydrofuran (40 mL) was added
a solution of NaOH (6.06 g, 151.0 mmol) in water (20 mL) and the
solution was heated at 60.degree. C. for 18 hr. The solution was
concentrated in vacuo and the aqueous residue was acidified to
pH=2.0. The resulting precipitate was filtered to give the desired
hydrochloride salt as a white solid (7.98 g, quantitative). MS
MH.sup.+ calculated for C.sub.22H.sub.22NSO.sub.5F.sub.- 3: 470,
found 470.
[0259] Part C: To a solution of the hydrochloride salt of Part B
(7.60 g, 15.0 mmol) in DMF (100 mL) were added
1-hydroxybenzotriazole (2.44 g, 18.0 mmol), N-methyl morpholine
(3.4 mL, 30.9 mmol) and O-tetrahydropyranyl hydroxylamine
hydrochloride (2.63 g, 22.5 mmol) followed by
1-3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (4.02
g, 21.0 mmol). The solution was stirred at ambient temperature for
96 hr. The solution was diluted with H.sub.2O (400 mL) and
extracted with ethyl acetate. The organic layer was washed with
saturated NaCl and dried over MgSO.sub.4, filtered and concentrated
in vacuo. Chromatography on silica eluting with 30% ethyl
acetate/hexane provided the desired tetrahydropyranyl-protected
hydroxamate as a white solid (5.93 g, 69%).
[0260] Part D: To a solution of tetrahydropyranyl-protected
hydroxamate of Part C (3.8 g, 6.7 mmol) in dioxane (100 mL) was
added 4 N HCl/dioxane (30 mL). The reaction was stirred at ambient
temperature for 2 hr, then the solution was concentrated in vacuo.
Trituration with diethyl ether afforded the title compound as a
white solid (3.33 g, 96%). MS MH.sup.+ calculated for
C.sub.22H.sub.23N.sub.2SO.sub.5F.sub.3: 485, found 485.
Preparative Example XI
Preparation of Resin II
[0261] Step 1: Attachment of Compound of Preparative Example IV to
Resin I.
[0262] A 500 mL round-bottomed flask was charged with of resin I
[Floyd et al., Tetrahedron Lett. 1996, 37, 8045-8048] (8.08 g, 9.7
mmol) and 1-methyl-2-pyrrolidinone (50 mL). A magnetic stirring bar
was added, and the resin slurry slowly stirred. A separate solution
of the compound of Part D, Preparative Example IV (5.58 g, 19.4
mmol) in 1-methyl-2-pyrrolidinone (35 mL) was added to the slurry
followed by addition of
benzotriazole-1-yl-oxy-tris-pyrrolidino-phosphonium
hexafluorophosphate (10.1 g, 19.4 mmol) in one portion. Once the
hexafluorophosphate salt had dissolved, 4-methylmorpholine (4.26
mL, 39 mmol) was added dropwise. The reaction slurry was stirred at
room temperature for 24 hr, then the resin was collected in a
sintered-disc funnel and washed with N,N-dimethylformamide,
methanol, methylene chloride and diethyl ether (3.times.30 mL each
solvent). The resin was dried in vacuo to yield 10.99 g
polymer-bound hydroxymate as a tan polymeric solid. Theoretical
loading on polymer was 0.91 mmol/g. FTIR microscopy showed bands at
1693 and 3326 cm.sup.-1 indicative of the hydroxamate carbonyl and
nitrogen-hydrogen stretches, respectively.
[0263] Step 2: Preparation of Resin III:
[0264] Reaction of Resin II with Nucleophiles
[0265] Resin II (50 mg, 0.046 mmol) was weighed into an 8 mL glass
vial, and a 0.5 M solution of a nucleophile in
1-methyl-2-pyrrolidinone (1 mL) was added to the vessel. In the
case of phenol and thiophenol nucleophiles, cesium carbonate (148
mg, 0.46 mmol) was added, and in the case of substituted piperazine
nucleophiles, potassium carbonate (64 mg, 0.46 mmol) was added. The
vial was capped and heated to 70 to 155.degree. C. for 24-48 hr,
then cooled to room temperature. The resin was drained and washed
with 1-methyl-2-pyrrolidinone, 1-methyl-2-pyrrolidinone/water
(1:1), water, 10% acetic acid/water, methanol, and methylene
chloride (3.times.3 mL each solvent).
[0266] Large Scale Preparation of Resin IIIa:
[0267] Resin II (5 g, 0.91 mmol) was weighed into an oven-dried
three-necked round bottom flask fitted with a temperature probe, an
overhead stirring paddle, and a nitrogen inlet. Anhydrous
1-methyl-2-pyrrolidinone (35 mL) was added to the flask followed by
ethyl isonipecotate (7.0 mL, 45.5 mmol). The resin slurry was
stirred slowly with the overhead stirrer, and the mixture was
heated to 80.degree. C. with a heating mantle for 65 hr. The flask
was thereafter cooled to room temperature.
[0268] The resin was collected in a sintered-disk glass funnel and
washed with N,N-dimethylformamide, methanol and methylene chloride
(3.times.30 mL each solvent). The resin was dried in vacuo to
provide 5.86 g of resin IIIa as off-white resin beads. The
theoretical loading of the polymer was 0.81 mmol/g. TFA cleavage
performed on 50 mg of resin Ea as described in step 3 yielded 10.4
mg of off-white solid spectroscopically indistinguishable from a
known sample.
[0269] Step 3: Cleavage of Hydroxamic Acids from the
Polymer-Support
[0270] Resin III was treated with a trifluoroacetic acid/water
mixture (19:1, 1 mL) for 1 hr at room temperature. During that
time, the resin became a deep red color. The resin was then drained
and washed with trifluoroacetic acid/water (19:1) and methylene
chloride (2.times.1 mL each solvent), collecting the combined
filtrates in a tared vial. The volatiles were removed in vacuo,
then a toluene/methylene chloride mixture (2 mL each) was added to
the residue. The mixture was again concentrated in vacuo. The
product was characterized by electrospray mass spectroscopy.
[0271] Step 4: Hydrolysis of Polymer-Bound Ester: Preparation of
Resin IVa
[0272] Resin IIIa (5.8 g, 4.5 mmol) was weighed into a three-necked
round bottomed flask fitted with an overhead stirring paddle.
1,4-Dioxane was added to the flask, and the resin slurry was
stirred for 15 min. Then, a 4 M solution of KOH (5 mL, 20 mmol) was
added, and the mixture was stirred for 44 hr. The resin was
thereafter collected in a sintered-disk glass funnel and washed
with dioxane/water (9:1), water, 10% acetic acid/water, methanol
and methylene chloride (3.times.30 mL each solvent). The resin was
dried in vacuo to yield 5.64 g of resin IVa as off-white polymer
beads. FTIR microscopy showed bands at 1732 and 1704 cm.sup.-1 and
a broad band from 2500-3500 cm.sup.-1. The theoretical loading of
the polymer-bound acid was 0.84 mmol/g.
Examples 1-45
[0273] The following compounds were prepared by parallel synthesis
(resin based synthesis, automated synthesis) using parallel
synthesis from Resin IVa as described previously in Preparative
Example XI the following compounds were prepared:
1 41 MS Example Amine R (M + H) 1 3,5-Dimethylpiperidine 42 508 2
1-(1-phenylethyl)-piperazine 43 585 3 1-(2-phenylethyl)-piperazine
44 585 4 1-(2-chlorophenyl)- piperazine 45 591 5
1-(4-methoxyphenyl)-2- methylpiperazine 46 585 6 1-(5-Chloro-2-
methylphenyl)piperazine 47 605 7 1-(2-methoxyphenyl)- piperazine 48
587 8 1-Acetylpiperazine 49 523 9 1-(2,4-Dimethylphenyl)-
piperazine 50 585 10 N-(2-hydroxyethyl)- piperazine 51 525 11
1-(Ethoxy-carbonylmethyl)- piperazine 52 567 12 1-(2-Fluorophenyl)-
piperazine 53 575 13 1-(2-Furoyl)- piperazine 54 575 14
1-(Cyclopentyl)-piperazine 55 549 15 1-(2-Propyl)- piperazine 56
523 16 N-(2-(1-Piperazino)- acetyl)pyrrolidine 57 592 17
1-(3-Dimethyl- aminopropyl)- piperazine 58 566 18
1-(2-Methoxyethyl)- piperazine 59 539 19 1-(2-Dimethyl-aminoethyl)-
piperazine 60 552 20 1-(2-Ethoxyphenyl)- piperazine 61 601 21
1-(4-Fluorphenyl)- piperazine 62 575 22 1-(2-Pyridyl)- piperazine
63 558 23 2-(1-piperazinyl)-pyrimidine 64 559 24 4-Piperazino-
acetophenone 65 599 25 1-(4-Nitrophenyl)- piperazine 66 602 26
1-(3,5-Dichloropyrid-4- yl)piperazine 67 626 27
4-(2-Methoxyphenyl)- piperidine 68 586 28 N-[2-Nitro-4-
(trifluoromethyl)- phenyl]piperazine 69 670 29
1-[3-(Trifluormethyl)-pyrid- 2-yl]- piperazine 70 626 30
cis-3,5-Dimethyl- morpholine 71 510 31 1-(2,4-Difluorphenyl)-
piperazine 72 593 32 1-(4-Pyridyl)- piperazine 73 558 33
1-(4-Trifluoromethyl- phenyl)-piperazine 74 625 34
1-Allylpiperazine 75 521 35 1-(2-Pyrazinyl)-piperazine 76 559 36
1-[3-Chloro-5- (trifluoromethyl)pyrid-2- yl)]piperazine 77 660 37
1-(2-(4-Morpholino)- ethyl)piperazine 78 594 38
3-Chlorophenyl-piperazine 79 591 39 4-(Hydroxymethyl)- piperidine
80 510 40 cis-2,6-Dimethyl-piperazine 81 509 41 3-Methylpiperidine
82 494 42 1-[4-(Trifluormethyl)- 2-pyrimidyl]- piperazine 83 627 43
1-[4-(Trifluormethyl)- 2-pyridyl]- piperazine 84 626 44
3,5-Dimethyl- piperidine 85 508 45 3,5-Dimethyl- piperidine 86
508
Examples 46-47
Step 12: Further Synthesis of Resin III
[0274] Into a 8 mL glass vial was placed resin II (200 mg, 0.18
mmol) and cesium carbonate (0.98 g mg, 3 mmol) (no cesium carbonate
used with piperidine and pyrrolidine nucleophiles). One mL of a 1.8
M solution of the amine nucleophile to be reacted in
1-methyl-2-pyrrolidinone (1.8 mmol) was added and the vial was
capped and heated to 100.degree. C. for 30 hr. Then the vessel was
cooled to room temperature, and the resin was drained and washed
with 1-methyl-2-pyrrolidinone, 1:1 1-methyl-2-pyrrolidinone/water,
water, 1:9 acetic acid/water, methanol and methylene chloride
(3.times.3 mL each solvent).
[0275] The following hydroxamic acids were synthesized from Resin
III with the indicated amines, followed by release from the polymer
using the reaction conditions in Step 3.
2 87 Example Amine R MS (M + H) 46 1-(2-Methoxyphenyl)- piperidine
88 475 47 4-(4-Methoxybenzoyl)- piperidine 89 503
Example 48
Preparation of
N-hydroxy-4-[[4-(4-methoxyphenoxy)phenyl]sulfonyl]-4-thiane-
carboxamide
[0276] 90
[0277] Step 1: Hydrolysis of methyl
4-[[4-(4-methoxyphenoxy)phenyl]sulfony- l]-4-thianecarboxylate. To
a solution of methyl 4-[[4-(4-methoxyphenoxy)-p-
henyl]sulfonyl]-4-thianecarboxylate (10.0 g, 31 mmol) dissolved in
tetrahydrofuran (150 mL) was added potassium trimethylsilanolate
(12.1 g) and stirred 2 hr. Water was added to the reaction mixture
and extracted with ethyl acetate (2.times.100 mL). The pH value of
the aqueous layer was adjusted to 2 with 2M hydrochloric acid and
extracted with ethyl acetate (2.times.100 mL). The latter organics
were washed with brine, dried over magnesium sulfate, filtered and
the solvent evaporated to afford a pale yellow solid (8.20 g).
[0278] Step 2: Loading on resin. The compound obtained in step 1
(4.0 g, 13.1 mmol) was dissolved in 1-methyl-2-pyrrolidinone (15
mL) and added to a suspension of resin I (6.0 g, 6.6 mmol;
Preparative Example XI) in 1-methyl-2-pyrrolidinone (40 mL). To
this solution were added pyBOP (6.85 g) and N-methylmorpholine (2.9
mL), and the mixture was stirred with overhead stirring 16 hr. The
resin was filtered and washed with dimethylformamide (3.times.50
mL), methanol (3.times.50 mL), dichloromethane (3.times.50 mL) and
ether (3.times.50 mL). The resin was dried in vacuo to provide
resin MT-III (6.79 g).
[0279] Step 3: Aryl fluoride displacement of resin MT-III. A
suspension of resin MT-III (200 mg, 0.17 mmol),
1-methyl-2-pyrrolidinone (2 mL), cesium carbonate (560 mg) and
4-methoxyphenyl (306 mg) were stirred at 105.degree. C. for 16 hr.
The reaction mixture was cooled and the resin filtered. The resin
was washed with dimethylformamide (3.times.5 mL), methanol
(3.times.5 mL), 10% aqueous acetic acid (3.times.5 mL), methanol
(3.times.5 mL) and dichloromethane (3.times.5 mL). To the resin was
added 95% aqueous trifluoroacetic acid and the reaction mixture was
agitated for 1 hr. The resin was drained and washed with
dichloromethane (2.times.1 mL). The solvent was evaporated. The
residue was purified by RPHPLC to provide
N-hydroxy-4-[[4-(4-methoxy-phenoxy)phenyl]sulfonyl]-4-t-
hianecarboxamide (17.9 mg) as a pale yellow oil.
Examples 49-50
[0280] The following hydroxamic acids were prepared by the method
of Example 48 using the appropriate amine.
3 91 MS (ES) Example R Amine m/z 49 4-(4-fluoro-benzoyl)
4-(4-fluorobenzoyl)- 507 piperidyl piperidine (M + H).sup.+ 50
4-(2-methoxy-phenyl) 4-(2-methoxyphenyl)- 491 piperidyl piperidine
(M + H).sup.+
Example 51
Preparation of
N-hydroxy-4-[[4-(4-methoxyphenoxy)phenyl]sulfonyl]-4-thiane-
carboxamide-1,1-dioxide
[0281] 92
[0282] Step 1: Oxidation of Resin MT-III. A suspension of resin
MT-III (2.0 g, 1.72 mmol), m-chloroperbenzoic acid (4.37 g) and
dichloromethane (25 mL) was stirred at room temperature for 20 hr.
The resin was filtered and washed with dichloromethane (3.times.25
mL), dimethylformamide (3.times.25 mL), methanol (3.times.25 mL),
1M aqueous sodium bicarbonate (2.times.25 mL), methanol (3.times.25
mL), dichloromethane (3.times.25 mL) and ether (3.times.25 mL). The
resin was dried in vacuo to afford resin MT-IV (2.16 g).
[0283] Step 2: Aryl fluoride displacement of resin MT-IV.
N-hydroxy-4-[[4-(4-methoxyphenoxy)-phenyl]sulfonyl]-4-thianecarboxamide
1,1-dioxide was prepared by the method of Example 48 using resin
MT-IV in the place of resin MT-III. ES (MS) m/z 473
(M+NH.sub.4).sup.+.
Example 52
[0284] The following hydroxamic acid was prepared by the method of
Example 51 using 4-(4-fluoro-benzoyl)-piperidine as the amine. MS
(ES) m/z 539 (M+H).sup.+. 93
Example 53
Preparation of
N-hydroxy-4-[[4-[4-[(3,5-dimethylpiperidyl)carbonyl]-piperi-
dyl]phenyl]sulfonyl]-4-thianecarboxamide
[0285] 94
[0286] Step 1: Aryl fluoride displacement of Resin MT-III. To a
suspension of resin MT-III (4.06 g, 3.4 mmol) in
1-methyl-2-pyrrolidinone (40 mL) was added ethyl isonipecotate
(5.25 mL), and the mixture was heated to 100.degree. C. for 16 hr.
The cooled reaction mixture was filtered and the resin was washed
with methanol (3.times.25 mL), dichloromethane (1.times.10 mL) and
ether (3.times.25 mL). The resin was dried in vacuo to afford resin
MT-V (4.21 g).
[0287] Step 2: Hydrolysis of resin MT-V. To a suspension of resin
MT-V (4.13 g) in tetrahydrofuran (20 mL) was added 4M aqueous
potassium hydroxide (10 mL) and stirred at room temperature for 5
days. The resin was filtered and washed with methanol (3.times.25
mL), dichloromethane (3.times.25 mL) and ether (3.times.25 mL). The
resin was dried in vacuo to afford resin MT-VI.
[0288] Step 3: Conversion to amide. To a suspension of resin MT-VI
(268 mg) in 1-methyl-2-pyrrolidinone (2 mL) were added
3,5-dimethyl-piperidine (299 .mu.L), pyBOP (587 mg) and
diisopropylethyl amine (393 .mu.L), and mixture was stirred 40 hr.
The resin was filtered and washed with dimethylformamide (3.times.2
mL), methanol (3.times.2 mL), 10% aqueous acetic acid (3.times.2
mL), methanol (3.times.2 mL), dichloromethane (3.times.2 mL) and
glacial acetic acid (1.times.2 mL). The resin was treated with 95%
aqueous trifluoroacetic acid (2 mL) and agitated 1 hr. The resin
was washed with dichloromethane (2 mL) and methanol (2 mL). The
filtrate was evaporated. The residue was purified by RPHPLC to
afford
N-hydroxy-4-[[4-[4-[(3,5-dimethylpiperidyl)carbonyl]piperidyl]phenyl]sulf-
onyl]-4-thianecarboxamide (7.5 mg) MS (ES) m/z 524 (M+H).sup.+.
Example 54
Preparation of
1,1-dimethylethyl-3,6-dihydro-4-[2-(trifluoromethyl)phenyl]- -1
(2H)-pyridinecarboxylate
[0289] 95
[0290] Part A: An oven-dried 1.0 liter flask fitted with a
thermometer and nitrogen inlet was charged with 55 mL of a 2 M
solution of lithium diisopropoylamide in tetrahydrofuran and 50 mL
of tetrahydrofuran. The flask was immersed in a dry ice/acetone
bath. When the temperature of the solution was less than
-70.degree. C., a solution of N-t-butoxycarbonylpiperidinone (20.0
g, 0.1 mole) in 100 mL tetrahydrofuran was added dropwise,
maintaining the temperature less than -65.degree. C. After complete
addition, the flask was stirred with cooling for 20 min. Then a
solution of N-trifluoromethanesulfonimide (38.2 g, 0.107 mole) was
added drop-wise maintaining the temperature less than -65.degree.
C. After complete addition, the dry ice/acetone bath was swapped
with an ice/water bath. The reaction was stirred overnight (about
18 hr), slowly warming to room temperature. After 16 hr, the
solvent was removed in vacuo, and the residue was purified by
column chromatography on neutral alumina, yielding 26.53 g of
product as a yellow oil. Electrospray mass spectroscopy showed m/z
332 (M+H).
[0291] Part B: A three-necked 15 mL round-bottom flask was charged
with the product from Part A (6 g, 18.1 mmol),
o-trifluorobenzeneboronic acid (4.94 g, 26 mmol), lithium chloride
(2.34 g, 55 mmol), 2 M sodium carbonate (26 mL, 52 mmol) and
ethylene glycol dimethyl ether (60 mL). Nitrogen was bubbled
through the solution for 10 min, then palladium
tetrakistriphenylphosphine (1.06 g, 0.92 mmol) was added. The
mixture was heated to reflux for 1.5 hr, then cooled to room
temperature. The solvent was removed in vacuo, then the residue was
partitioned between 100 mL of methylene chloride and 100 mL of 2 M
sodium carbonate with 3 mL concentrated ammonium hydroxide. The
aqueous layer was extracted with an additional 100 mL methylene
chloride, then the combined organic layers were dried over
magnesium sulfate and concentrated to give 8.42 g of crude product
as a dark brown oil. Purification via flash column chromatography
(10% ethyl acetate3/hexanes) yielded 2.76 g of pure product as a
yellow oil. Electrospray mass spectroscopy showed m/z 328
(M+H).
Example 55
Preparation of
1,2,3,6-tetrahydro-4-[2-trifluoromethyl)phenyl]pyridine
[0292] 96
[0293] The title compound of Example 54 (300 mg, 0.92 mmol) was
dissolved in methylene chloride (5 mL) in a 15 mL round-bottom
flask, and 5 mL of trifluoroacetic acid was added dropwise. After
15 min, the solvent was removed in vacuo, and the residue
partitioned between 20 mL of ethyl acetate and 20 mL of 2 M sodium
carbonate. The organic layer was washed with additional 2 M sodium
carbonate, dried over magnesium carbonate and concentrated in vacuo
to yield 195 mg of pure product as a colorless oil. Electrospray
mass spectroscopy showed m/z 228 (M+H).
Example 56
Preparation of 4-[2-(trifluoromethyl)phenyl]piperidine
[0294] 97
[0295] Part A: A solution of the title compound of Example 54 (2.3
g, 7 mmol) in 20 mL ethanol was added to a hydrogenation flask
containing 1 g of 4% palladium on carbon (0.38 mmol). The mixture
was placed under 100 PSI hydrogen and heated to 50.degree. C. for 5
hr. Then the mixture was cooled to room temperature and filtered
through Celite. The filtrate was concentrated in vacuo to give 2.27
g of pure product as a colorless oil. Electrospray mass
spectroscopy showed m/z 330 (M+H).
[0296] Part B: The product from Part A above (2.24 g, 6.8 mmol) was
dissolved in 100 mL methylene chloride, and 100 mL of
trifluoroacetic acid was added dropwise. After 15 min, the solvent
was removed in vacuo, and the residue partitioned between 100 mL of
ethyl acetate and 100 mL of 2 M sodium carbonate. The organic layer
was washed with additional 2 M sodium carbonate, dried over
magnesium carbonate and concentrated in vacuo to yield 1.12 g of
pure product as a colorless oil. Electrospray mass spectroscopy
showed m/z 230 (M+H).
Example 57
General Description for Preparation of Hydroxamic Acids via Aryl
Fluoride Displacement with Amines
[0297] Part A: A 2 dram vial was charged with aryl fluoro compound
of Preparative Example IV (170 mg, 0.44 mmol), 1 ml of
2-methylpyrrolidinone, cesium carbonate (360 mg, 1.1 mmol) and 0.66
mmol of an amine. A small magnetic stirring bar was added, then the
vial was capped and placed in a Pierce Reacti-therm.TM. at
115.degree. C. The reaction progress was followed by analytical
HPLC. When the reaction was greater than 90% complete, the vial was
cooled to room temperature. The reaction mixture was diluted with 5
mL of water, then 1.2 mL of 5% hydrogen chloride/water was added
dropwise. Then, the entire mixture was poured onto a column of
Celite. The column was washed exhaustively with ethyl acetate
(30-40 mL) and the filtrate was collected and concentrated to give
the crude products.
[0298] Part B: The product from above was dissolved in 2 mL
1,4-dioxane and 2 mL of methanol in a 4 dram vial with a small
magnetic stirring bar. A solution of 4 N hydrogen chloride in
1,4-dioxane was carefully added to the reaction, and the mixture
was stirred for 2 hr. Then the solvent was removed in vacuo and the
residue purified by preparative reversed-phase HPLC.
Examples 58-60
[0299] The following hydroxamic acids were prepared using the
method described above in Example 57 with the indicated amine as
the starting material.
4 98 m/z from electrospray mass Example amine R spectroscopy 58
Product of Example 56 99 513.3 (M + H) 59 Product of Example 55 100
511.2 (M + H) 60 4-(2-keto- benzimid- azolinyl)- piperidine 101 501
(M + H)
Examples 61-69
[0300] Using the procedures outlined in Examples 54, 55, and 57 and
other methods outlined above, the following analogs are made from
the indicated boronic acid:
5 102 Example Boronic acid R 61 103 104 62 105 106 63 107 108 64
109 110 65 111 112 66 113 114 67 115 116 68 117 118 69 119 120
Example 70
Preparation of
4-[[4-[4-[(3,5-dimethyl-1-piperidinyl)carbonyl]-1-piperidin-
yl]-phenyl]sulfonyl]-N-hydroxy-1-(2-methoxyethyl)-4-piperidinecarboxamide,
monohydrochloride
[0301] 121
[0302] Part A: To a solution of isonipecotic acid (5.8 g, 44.9
mmol) in water (200 mL) was added sodium carbonate (4.62 g, 44.9
mmol) followed by the drop-wise addition of
di-tert-butyl-dicarbonate (10.1 g, 46.3 mmol) in dioxane (40 mL).
After 4 hr, the solvent was concentrated in vacuo and the solution
was extracted with ethyl ether. The aqueous layer was acidified
with 3N hydrochloric acid to pH=2. The solution was extracted with
ethyl ether and the organic layer was washed with saturated aqueous
sodium chloride and dried over magnesium sulfate. Concentration in
vacuo provided N-Boc-isonipecotic acid as a white solid (9.34 g,
90%).
[0303] Part B: To a solution of the N-Boc-isonipecotic acid of part
A (1.0 g, 4.37 mmol) in dichloromethane (10 mL) was added
1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (853
mg, 4.45 mmol), 1-hydroxybenzotriazole hydrate (620 mg, 4.59 mmol)
3,5-dimethylpiperdine (0.67 mL, 5.03 mmol) and
diisopropylethylamine (1.67 mL, 9.61 mmol) and was stirred for 21
hr. The solution was concentrated in vacuo. The residue was diluted
with ethyl acetate and washed with 1M hydrochloric acid, saturated
sodium bicarbonate and saturated aqueous sodium chloride and dried
over sodium sulfate. Concentration in vacuo provided the amide as a
clear colorless oil (1.21 g, 89%).
[0304] Part C: To a solution of the amide of part B (1.20 g, 3.84
mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (5
mL) and the solution was stirred for 1 hr. Concentration in vacuo
provided an oil which was added directly to a solution of the
compound of Preparative Example VII, Part A (956 mg, 2.56 mmol) in
dimethylacetamide (10 mL). Cesium carbonate (2.92 g, 8.96 mmol) was
added and the solution was heated to 100.degree. C. for 18 hr. The
solution was partitioned between ethyl acetate and water and the
organic layer was washed with water and saturated sodium chloride
and dried over sodium sulfate. Concentration in vacuo provided the
phenylamine as an oil (1.53 g, 68%). MS(CI) MH.sup.+ calculated for
C.sub.30H.sub.47N.sub.3O.sub.6S: 578, found 578.
[0305] Part D: To a solution of the phenylamine of part C (1.5 g,
2.6 mmol) in ethanol (9 mL) and tetrahydrofuran (9 mL) was added
sodium hydroxide (1.02 g, 26 mmol) in water (5 mL) and the solution
was heated to 60.degree. C. for 20 hr. The solution was
concentrated and the residue was diluted with water and acidified
to pH=3 with 3N hydrochloric acid. Vacuum filtration provided the
acid as a beige solid (500 mg, 33%). MS(CI) MH.sup.+ calculated for
C.sub.28H.sub.43N.sub.3O.sub.6S: 550, found 550.
[0306] Part E: To a solution of the acid of part D (492 mg, 0.84
mmol) in N,N-dimethylformamide (10 mL) was added
1-hydroxybenzotriazole hydrate (136 mg, 1.01 mmol),
4-methylmorpholine (0.46 mL, 4.20 mmol), and O-tetrahydropyranyl
hydroxylamine (147 mg, 1.26 mmol). After 1 hr,
1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (225
mg, 1.18 mmol) was added and the solution was stirred for 72 hr at
ambient temperature. The solution was partitioned between ethyl
acetate and water. The organic layer was washed with water and
saturated sodium chloride and dried over sodium sulfate.
Concentration in vacuo provided the protected hydroxamate as an oil
(524 mg, 96%). MS(CI) MH.sup.+ calculated for
C.sub.33H.sub.51N.sub.4O.sub.7S: 649, found 649.
[0307] Part F: To a solution of the protected hydroxamate of part E
(514 mg, 0.79 mmol) in 1,4-dioxane (10 mL) was added 4M
hydrochloric acid in dioxane (10 mL) and the solution was stirred
for 1.5 hr. The solution was concentrated in vacuo and trituration
(ethyl ether) provided the title compound as a white solid (360 mg,
76%). MS(CI) MH.sup.+ calculated for
C.sub.28H.sub.44N.sub.4O.sub.6S: 565, found 565. HRMS calculated
for C.sub.28H.sub.44N.sub.4O.sub.6S: 565.3060, found 565.3070.
Analytical calculation for C.sub.28H.sub.44N.sub.4O.sub.6S
2HCl:2H.sub.2O: C, 49.92; H, 7.48; N, 8.32; S, 4.76; Cl, 10.52.
Found: C, 49.41; H, 7.55; N, 7.85; S, 4.53; Cl, 10.78.
Example 71
Preparation of
4-[[4-[4-[(3,5-dimethyl-1-piperidinyl)carbonyl]-1-piperidin-
yl]-phenyl]sulfonyl]-N-hydroxy-1-(2-methoxyethyl)-4-piperidinecarboxamide
[0308] 122
[0309] A solution of the hydroxamate of Example 70, part F (50 mg,
0.08 mmol) in water (2 mL) was neutralized with saturated sodium
bicarbonate. The aqueous solution was extracted with ethyl acetate.
Concentration in vacuo provided the hydroxamate free base as an
orange solid (35 mg, 75%).
Example 72
Preparation of
rel-4-[[4-[4-[[(3R,5R)-3,5-dimethyl-1-piperidinyl]carbonyl]-
-1-piperidinyl]phenyl]sulfonyl]-N-hydroxy-4-piperidinecarboxamide,
monohydrochloride
[0310] 123
[0311] Part A: To a solution of the N-Boc-isonipecotic acid of
Example 70, Part A (1.0 g, 4.37 mmol) in dichloromethane (10 mL)
was added 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide
hydrochloride (853 mg, 4.45 mmol), 1-hydroxybenzotriazole hydrate
(620 mg, 4.59 mmol) 3,5-dimethylpiperdine (0.67 mL, 5.03 mmol) and
diisopropylethylamine (1.67 mL, 9.61 mmol) and was stirred for 21
hr. The solution was concentrated in vacuo. The residue was diluted
with ethyl acetate and washed with 1M hydrochloric acid, saturated
sodium bicarbonate and saturated sodium chloride and dried over
sodium sulfate. Concentration in vacuo provided the amide as a
clear colorless oil (1.4 g, quantitative yield).
[0312] Part B: To a solution of the amide of part A (1.4 g, 4.49
mmol) in dioxane (10 mL) was added 4M hydrochloric acid in dioxane
(10 mL) and the solution was stirred for 1 hr. Concentration in
vacuo provided a solid that was added directly to a solution of the
compound of Preparative Example II, Part D, (1.24 mg, 2.99 mmol) in
dimethylacetamide (10 mL). Cesium carbonate (3.42 g, 10.5 mmol) was
added and the solution was heated to 100.degree. C. for 20 hr. The
solution was partitioned between ethyl acetate and water and the
organic layer was washed with water and saturated sodium chloride
and dried over sodium sulfate. Concentration in vacuo provided the
phenylamine as a yellow solid (1.90 g, quantitative yield). MS(CI)
MH.sup.+ calculated for C.sub.32H.sub.49N.sub.3O.sub.7S: 620, found
620.
[0313] Part C: To a solution of the phenylamine of part B (1.9 g,
3.0 mmol) in ethanol (10 mL) and tetrahydrofuran (10 mL) was added
sodium hydroxide (1.2 g, 30 mmol) in water (5 mL) and the solution
was heated to 60.degree. C. for 20 hr. The solution was
concentrated and the residue was diluted with water and acidified
to pH=1 with 3N hydrochloric acid. The solution was extracted with
ethyl acetate and washed with 1M hydrochloric acid and saturated
sodium chloride and dried over magnesium sulfate. Concentration in
vacuo provided the acid as a yellow oil (1.9 g, quantitative
yield). MS(CI) MH.sup.+ calculated for
C.sub.30H.sub.45N.sub.3O.sub.7S: 592, found 592.
[0314] Part D: To a solution of the acid of part C (1.87 g, 3.00
mmol) in N,N-dimethylformamide (10 mL) was added
1-hydroxybenzotriazole hydrate (486 mg, 3.6 mmol),
4-methylmorpholine (1.65 mL, 15 mmol), and O-tetrahydropyranyl
hydroxylamine (526 mg, 4.5 mmol). After 1 hr,
1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (805
mg, 4.2 mmol) was added and the solution was stirred for 18 hr at
ambient temperature. The solution was partitioned between ethyl
acetate and water. The organic layer was washed with water and
saturated sodium chloride and dried over sodium sulfate.
Chromatography (on silica, ethyl acetate/hexane) provided the
protected hydroxamate as an oil (1.63 g, 79%).
[0315] Part E: To a solution of the protected hydroxamate of part D
(1.61 g, 2.33 mmol) in dioxane (10 mL) was added 4M hydrochloric
acid in dioxane (10 mL) and the solution was stirred for 45 min.
The solution was concentrated in vacuo and trituration (ethyl
ether) a white solid. Reverse phase chromatography (on silica,
acetonitrile/water (hydrochloric acid)) produced fractions A, B, C
and D. Concentration in vacuo of fraction A provided the title
compound as a white solid (59 mg). MS(CI) MH.sup.+ calculated for
C.sub.25H.sub.38N.sub.4O.sub.5S: 507, found 507.
Example 73
Preparation of rel-1,1-dimethylethyl
4-[[4-[4-[[(3R,5R)-3,5-dimethyl-1-pip-
eridinyl]carbonyl]-1-piperidinyl]-phenyl]sulfonyl]-4-[(hydroxyamino)-carbo-
nyl]-1-piperidinecarboxylate
[0316] 124
[0317] From the reverse phase chromatography of Example 72, Part E,
fraction C was concentrated in vacuo to provide the title compound
as a white solid (49 mg). MS(CI) MH.sup.+ calculated for
C.sub.30H.sub.46N.sub.4O.sub.7S: 607, found 607.
Example 74
Preparation of
rel-4-[[4-[4-[[(3R,5S)-3,5-dimethyl-1-piperidinyl]carbonyl]-
-1-piperidinyl]phenyl]sulfonyl]-N-hydroxy-4-piperidinecarboxamide,
monohydrochloride
[0318] 125
[0319] From the reverse phase chromatography of Example 72, Part E,
fraction B was concentrated in vacuo to provide the title compound
as a white solid (198 mg). MS(CI) MH.sup.+ calculated for
C.sub.25H.sub.38N.sub.4O.sub.5S: 507, found 507.
Example 75
Preparation of rel-1,1-dimethylethyl
4-[[4-[4-[[(3R,5S)-3,5-dimethyl-1-pip-
eridinyl]carbonyl]-1-piperidinyl]-phenyl]sulfonyl]-4-[(hydroxyamino)-carbo-
nyl]-1-piperidinecarboxylate
[0320] 126
[0321] From the reverse phase chromatography of Example 72, Part E,
fraction D was concentrated in vacuo to provide the title compound
as a white solid (242 mg). MS(CI) MH.sup.+ calculated for
C.sub.30H.sub.46N.sub.4O.sub.7S: 607, found 607.
Example 76
Preparation of
4-[[4-[4-[(2,3-dihydro-1H-indol-1-yl)carbonyl]-1-piperidiny-
l]-phenyl]sulfonyl]-N-hydroxy-1-(2-methoxyethyl)-4-piperidinecarboxamide,
monohydrate
[0322] 127
[0323] Part A: To a solution of the N-Boc-isonipecotic acid of
Preparative Example I, Part B (750 mg, 3.27 mmol) in
dichloromethane (3 mL) was added
2-chloro-4,6-dimethoxy-1,3,5-triazine (564 mg, 3.21 mmol). The
solution was cooled to 0.degree. C. and 4-methylmorpholine (0.35
mL, 3.21 mmol) was added. After 2 hr, indoline (0.36 mL, 3.21 mmol)
was added and the solution was stirred for 22 hr at ambient
temperature. The solution was concentrated in vacuo. The residue
was diluted with ethyl acetate and washed with 1M hydrochloric
acid, saturated sodium bicarbonate and saturated sodium chloride
and dried over sodium sulfate. Concentration in vacuo provided the
amide as a pink solid (940 mg, 89%).
[0324] Part B: To a solution of the amide of part A (935 g, 2.83
mmol) in 1,4-dioxane (10 mL) was added 4M hydrochloric acid in
dioxane (10 mL) and the solution was stirred for 1 hr.
Concentration in vacuo provided an oil which was added directly to
a solution of the compound of Preparative Example VII, Part A, (705
mg, 1.89 mmol) in dimethylacetamide (10 mL). Cesium carbonate (2.15
g, 6.61 mmol) was added and the solution was heated to 110.degree.
C. for 18 hr. The solution was partitioned between ethyl acetate
and water and the organic layer was washed with water and saturated
sodium chloride and dried over sodium sulfate. Concentration in
vacuo provided the phenylamine as an orange oil (893 mg, 81%).
MS(CI) MH.sup.+ calculated for C.sub.31H.sub.41N.sub.3O.sub.6S:
584, found 584.
[0325] Part C: To a solution of the phenylamine of part B (885 g,
1.52 mmol) in ethanol (10 mL) and tetrahydrofuran (10 mL) was added
sodium hydroxide (607 mg, 15.2 mmol) in water (5 mL) and the
solution was heated to 60.degree. C. for 20 hr. The solution was
concentrated and the residue was diluted with water and acidified
to pH=1 with 3N hydrochloric acid producing a solid. Vacuum
filtration provided the acid as a beige solid (475 g, 53%). MS(CI)
MH.sup.+ calculated for C.sub.29H.sub.37N.sub.3O.sub- .6S: 556,
found 556.
[0326] Part D: To a solution of the acid of part C (465 g, 0.79
mmol) in N,N-dimethylformamide (10 mL) was added
1-hydroxybenzotriazole hydrate (128 mg, 0.95 mmol),
4-methylmorpholine (0.43 mL, 3.95 mmol), and O-tetrahydropyranyl
hydroxylamine (139 mg, 1.18 mmol). After 1 hr,
1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (212
mg, 1.10 mmol) was added and the solution was stirred for 18 hr at
ambient temperature. The solution was partitioned between ethyl
acetate and water. The organic layer was washed with water and
saturated sodium chloride and dried over sodium sulfate.
Chromatography (on silica, ethyl acetate/methanol) provided the
protected hydroxamate as a yellow oil (305 mg, 60%). MS(CI)
MH.sup.+ calculated for C.sub.34H.sub.46N.sub.4O.sub.7S: 655, found
655.
[0327] Part E: To a solution of the protected hydroxamate of part D
(300 mg, 0.46 mmol) in dioxane (5 mL) was added 4M hydrochloric
acid in dioxane (5 mL) and the solution was stirred for 2 hr. The
resulting solid was collected by vacuum filtration. Washing with
ethyl ether provided the title compound as a white solid (260 mg,
94%). MS(CD) MH.sup.+ calculated for
C.sub.29H.sub.34N.sub.4O.sub.6S: 571, found 571.
[0328] The following compounds were prepared by parallel synthesis
(resin based synthesis, automated synthesis) procedures utilizing
reactions such as acylation and nucleophilic displacement:
Example 77
[0329] 128
Example 78
[0330] 129
Example 79
[0331] 130
Examples: 80-118
[0332]
6 131 Example R.sub.1R.sub.2NH Amine MS (ES) m/z 80 132 Ethyl amine
592 (M + H) 81 133 3-(Aminomethyl) pyridine 655 (M + H) 82 134
Imidazole 615 (M + H) 83 135 3-Amino-1-propanol 622 (M + H) 84 136
Histamine 658 (M + H) 85 137 2-Thiophene methyl amine 660 (M + H)
86 138 Morpholine 634 (M + H) 87 139 2-(Aminomethyl) pyridine 655
(M + H) 88 140 4-(Aminomethyl) pyridine 655 (M + H) 89 141
Ethanolamine 608 (M + H) 90 142 N,N,N-Trimethyl ethylenediamine 649
(M + H) 91 143 1-Methylpiperazine 647 (M + H) 92 144 N,N-Dimethyl
ethylenediamine 635 (M + H) 93 145 Piperazine 633 (M + H) 94 146
Thiomorpholine 650 (M + H) 95 147 N-Propylcyclopropne methylamine
660 (M + H) 96 148 (Aminomethyl) cyclopropane 618 (M + H) 97 149
Dimethylamine 592 (M + H) 98 150 Diethylamine 620 (M + H) 99 151
Piperidine 632 (M + H) 100 152 (R)-(-)-2-Pyrrolidine methanol 648
(M + H) 101 153 Pyrrolidine 618 (M + H) 102 154
1-(2-(2-Hydroxyethoxy) ethyl)piperazine 721 (M + H) 103 155
Isonipecotamide 675 (M + H) 104 156 2-(2-Aminoethoxy) ethanol 652
(M + H) 105 157 3,3'-Iminobis(N,N- dimethylpropylamine) 734 (M + H)
106 158 Bis(2-Methoxy ethyl)amine 680 (M + H) 107 159 4-Hydroxy
piperidine 648 (M + H) 108 160 N-(Carboethoxy methylpiperazine 719
(M + H) 109 161 1-(2-Morpholinoethyl) piperazine 746 (M + H) 110
162 1-(2-Methoxyethyl) piperazine 691 (M + H) 111 163 1-(2-
Dimethylaminoethyl) piperazine 704 (M + H) 112 164
2-Methoxyethylamine 622 (M + H) 113 165 2,2,2-Trifluoroethyl amine
646 (M + H) 114 166 1,2,4-Triazole 616 (M + H) 115 167 Methoxyamine
594 (M + H) 116 168 Ethyl isonipecotate 704 (M + H) 117 169
2-Pyrrolidinone 632 (M + H) 118 170 Isonipecotic acid 676 (M +
H)
Example 119
Preparation of
[0333] 171
[0334] Part A. Preparation of aryl fluoride. To a solution of ethyl
4-[(4-fluorophenyl)sulfonyl]-1-(2-methoxyethyl)-4-piperidinecarboxylate
(57 mmol) in dioxane (90 mL) and water (45 mL) was added LiOH (4.8
g, 3.5 eq). The mixture was stirred at 60.degree. C. overnight,
cooled to room temperature, and concentrated in vacuo. The aqueous
layer was treated with concentrated HCl until the pH was
approximately 4. The solid was collected and dried. Next, the solid
(47.8 mmol) in DMF (100 mL) was added NMM (26.2 mL, 239 mmol) and
(benzotriazol-1-yl)-N,N,N',N'-bis(tetra- methylene)uronium
tetrafluoroborate (32.3 g, 62.1 mmol). The mixture was stirred at
room temperature for 15 min, and O-(tetrahydro-211-pyran-2-yl)-
hydroxylamine (6.71 g, 57.32 mmol) was then added. After 48 hr at
room temperature, the mixture was quenched with sat.
NH.sub.4.sup.+Cl.sup.-, and then extracted with CH.sub.2Cl.sub.2
three times. The combined organic layer was dried and concentrated
in vacuo. The residue was purified over SiO.sub.2 using
hexane/CH.sub.2Cl.sub.2 and then CH.sub.2Cl.sub.2/MeOH to give 20 g
of protected hydroxyamide as an orange oil.
[0335] Part B. Aryl fluoride displacement. A solution of the aryl
fluoride from Part A (0.45 mmol), Cs.sub.2CO.sub.3 (1.35 mmol, 3
eq), and 4-(4-chlorobenzoyle)piperidine (Maybridge Chemical Co.,
England, 0.67 mmol, 1.5 eq) in DMSO (1 mL) was heated to
110.degree. C. for 18-48 hr. The mixture was then cooled, dissolved
in saturated aq. NH.sub.4.sup.+Cl.sup.- (5 mL), and extracted with
dichloromethane (3.times.3 mL). The combined organic layer was
blown down. The crude product was purified by RPHPLC (eluting with
10% to 90% acetonitrile/water), and the pure fractions were
combined and concentrated.
[0336] Part C. Conversion of the THP hydroxamic acid of Part B to
the hydroxamic acid. The residue from Part B was dissolved in 2 mL
of 4M HCl and 1 mL of MeOH, stirred at room temperature for 1 h,
and then blown down. THEO M+H=564.1935; Observed: HI RES
M+H=564.1949.
Example 120
Preparation of
[0337] 172
[0338] Part A. Preparation of 4-(4-n-propylbenzoyl)piperidine. To a
solution of magnesium (147 mmol, 5 eq) in 40 mL of THF at 0.degree.
C. was added 1-bromo-4-(n-propyl)benzene (88.24 mmol, 3 eq). The
solution was allowed to warm to room temperature over approximately
3 hr. The weinreb amide (29.4 mmol, 1 eq) having the following
structure: 173
[0339] was added, and then the mixture was stirred at room
temperature for 18 hr. The mixture was quenched with saturated
NH.sub.4.sup.+Cl.sup.-, and then extracted with CH.sub.2Cl.sub.2
three times. The combined organic layer was washed with saturated
NH.sub.4.sup.+Cl.sup.-, dried over MgSO.sub.4, and concentrated in
vacuo. The residue was purified over 70 g of SiO.sub.2, eluting
with ethylacetate:hexanes (1:10) to ethylacetate:hexanes (1:3). The
piperidine was dissolved in 20 mL of CH.sub.2Cl.sub.2 and 20 mL of
trifluoroacetate. The resulting mixture was stirred at room
temperature for 1 hr, and then concentrated in vacuo. The residue
was treated with 5% NaOH until a solid precipitated out. The solid
was collected and then dissolved in dichloromethane, dried, and
concentrated in vacuo. The residue was recrystallized in MeOH/Ether
to give 5.07 g of 4-(4-n-propylbenzoyl)piperidine.
[0340] Part B. Aryl fluoride displacement. A solution of the aryl
fluoride (0.45 mmol., as prepared in Part A of the preceding
example), Cs.sub.2CO.sub.3 (1.35 mmol, 3 eq), and the
4-(4-n-propylbenzoyl)piperidi- ne prepared in Part A above (0.65
mmol, 1.5 eq) in DMSO (1 mL) was heated to 110.degree. C. for 18-48
hr. The mixture was cooled, dissolved in saturated aqueous
NH.sub.4.sup.+Cl.sup.- (5 mL), and extracted with dichloromethane
(3.times.3 mL). The combined organic layer was blown down. The
crude product was purified by RPHPLC (eluting with 10% to 90%
acetonitrile/water), and the pure fractions were combined and
concentrated.
[0341] Part C. Conversion of the THP hydroxamic acid of Part B to
the hydroxamic acid. The residue from Part C was dissolved in 2 mL
of 4M HCl and 1 mL of MeOH, stirred at room temperature for 1 h,
and then blown down. Theo: M+H=572.2794; Observed: Hi Res
M+H=572.2755;
Example 121
Preparation of
[0342] 174
[0343] Part A: 175
[0344] To a solution of n-butylthiophene (Lancaster, 5.0 g, MW
140.26, 1.1 eq) in tetrahydrofuran (80 ml) at 0.degree. C. was
dripped in 1.6 M n-butyllithium in hexanes (Aldrich, 24 ml, 1.2
eq). The mixture stirred at 0.degree. C. for 0.5 hr under N.sub.2.
The reaction vessel was then cooled to -78.degree. C., and a
solution of the weinreb amide (shown in the reaction above) in
tetrahydrofuran (30 ml) was slowly added. The dry ice bath was
removed, and the reaction was allowed to warm to room temperature.
After 3 hr, the conversion was complete. The reaction was quenched
with water (50 ml), and the organic layer was removed in vacuo.
More water (100 ml) was added, and the mixture was extracted with
diethylether (3.times.100 ml). The organic layers were washed with
water (2.times.) and brine (1.times.), dried over Na.sub.2SO.sub.4,
and concentrated to afford a brown oil that was chromatographed
(ethylacetate:hexanes, 1:9) to afford 7.5 g of a pale yellow solid
(67% crude yield). .sup.1H NMR showed the desired compound.
[0345] Part B: 176
[0346] To a solution of Compound I (7.4 g, MW 351.50, 1.0 eq) in
acetonitrile (10 ml) was added 4 N HCl in dioxane (Pierce, 40 ml).
After 1 hr, the solvent was evaporated, and the residue was
slurried in diethylether to afford a white solid that was collected
and dried for 5.8 g (97% yield). .sup.1H NMR showed the desired
Compound II.
[0347] Part C: 177
[0348] To a solution of Compound II (2.1 g, MW 287.85, 1.5 eq) in
dimethylsulfoxide (Aldrich, 15 ml) was added CsCO.sub.3 (Aldrich,
6.4 g, MW 325.8, 4.0 eq). After stirring for 5 min, Compound III
(2.0 g, MW 401.49, 1.0 eq) was added, and the mixture was stirred
at 90.degree. C. for 24 hr. The mixture was then diluted with water
(15 ml), and extracted with ethylacetate (3.times.100 ml). The
organic layer was washed with water (1.times.), washed with brine
(2.times.), dried over Na.sub.2SO.sub.4, and concentrated to a
crude brown solid which was recrystallized from hot methanol for
1.83 g of an orange crystalline solid (59% yield). .sup.1H NMR
showed the desired Compound IV.
[0349] Part D: 178
[0350] To a solution of Compound IV (1.8 g, MW 632.88, 1.0 eq) in
methylene chloride (5 ml) was added trifluoroacetic acid (10 ml).
The mixture was stirred for 4 hr at room temperature. The mixture
was then concentrated to 1/3 volume, and diethylether was added to
afford a solid, which was collected and dried for 1.4 g tan solid
(88% yield). .sup.1H NMR and LCMS showed the desired Compound
V.
[0351] Part E: 179
[0352] To a solution of Compound V (1.3 g, 2.2 mmol) in
N,N-dimethylformamide (8 ml) was added triethylamine (Aldrich, 1.2
ml, 8.8 mmol), followed by N-hydroxybenzotriazole hydrate (Aldrich,
0.6 g, 4.4 mmol), O-tetrahydro-2H-pyran-2-yl)hydroxylamine (0.4 g,
3.3 mmol), and, lastly,
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (Sigma,
0.9 g, 4.8 mmol). The mixture was stirred for 2 days at room
temperature. The mixture was diluted with water (10 ml), and
extracted with ethylacetate (3.times.75 ml). The organic layers
were combined, and washed with a saturated sodium bicarbonate
solution (1.times.1 50 ml) and brine (1.times.150 ml). The organic
layer was then dried over Na.sub.2SO.sub.4, and concentrated to
afford an orange foam that was recrystallized from methanol to
afford a pale yellow solid (1.2 g, 80% yield). .sup.1H NMR and LCMS
showed the desired Compound VI.
[0353] Part F: 180
[0354] The Compound VI (1.2 g, 1.8 mmol) was treated with methanol
(0.5 ml) and 4 N HCl in dioxane (5 ml) for 1 hr. The solvents were
concentrated to 1/3 the volume via an N.sub.2 stream. Diethylether
was then added to the residue to afford a solid that was collected
and dried to a white solid (1.0 g, 91% yield). .sup.1H NMR showed
the desired Compound VII. HRMS confirmed this finding (theo M=H
592.2515, observed: 592.2498).
Example 122
Preparation of
[0355] 181
[0356] Part A: 182
[0357] To an ice-cold suspension of isopropylphosphonium iodide
(Aldrich, 40.7, MW 432.29, 3.0 eq) at 0.degree. C. in
tetrahydrofuran (240 ml) was slowly added n-butyllithium (Aldrich,
1.6 M, 58.9 ml, 3.0 eq). After 1 hr, a solution of
5-bromo-2-thiophene carboxaldehyde (Aldrich, 6.0 g, MW 191.05, 1.0
eq) in tetrahydrofuran (60 ml) was added in one shot. The ice bath
was removed, and the mixture warmed to ambient temperature and
stirred 2.5 hours. The reaction was quenched with water (110 ml)
followed by 1 N HCl (110 ml). An emulsion developed that was
filtered through a coarse frit funnel. The filtrate was separated
and the organic was washed with brine (200 ml), dried over
Na.sub.2SO.sub.4, and concentrated to afford a black oil.
Purification on silica gel (ethyl acetate/hexanes) gave 3.4 g of a
yellow oil (50% yield). .sup.1H NMR showed desired Compound I.
[0358] Part B: 183
[0359] To a solution of Compound I (2.89 g, MW 217.13, 1.5 eq) in
tetrahydrofuran (25 ml) at -40.degree. C. was dripped 2.0M
isopropylmagnesium chloride in tetrahydrofuran (Aldrich, 6.9 ml,
1.55 eq). The mixture was stirred at -40.degree. C. for 1.5 hr
under N.sub.2. A solution of the weinreb amide (shown in the above
reaction) in tetrahydrofuran (30 ml) was quickly added. The dry ice
bath was removed, and the mixture was allowed to warn to room
temperature and stirred overnight. The reaction was quenched with 1
N HCl (25 ml), followed by water (25 ml). The organic layer was
removed in vacuo. The aqueous residue was extracted with
diethylether (3.times.100 ml). The organic layers were washed with
water (2.times.) and brine (1.times.), dried over Na.sub.2SO.sub.4,
and concentrated to afford a brown oil that was slurried with
hexanes. A solid formed, which was subsequently filtered to afford
1.9 g of gray solid (61% crude yield). .sup.1H NMR showed the
desired Compound II.
[0360] Part C: 184
[0361] To Compound II (1.9 g, MW 349.49, 1.0 eq) was added 4 N HCl
in dioxane (Pierce, 10 ml). After 1 hr, the solvent was evaporated,
and the residue was slurried in diethylether to afford a gray solid
that was collected and dried for 1.4 g (93% yield). .sup.1H NMR
showed the desired Compound III.
[0362] Part D: 185
[0363] To a solution of Compound III (1.4 g, MW 285.83, 1.5 eq) in
dimethylsulfoxide (Aldrich, 10 ml) was addded CsCO.sub.3 (Aldrich,
4.0 g, MW 325.8, 4.0 eq). After 5 min, Compound IV (1.3 g, MW
401.49, 1.0 eq) was added, and the reaction was stirred at
100.degree. C. for 24 hr. The mixture was then diluted with water
(15 ml), and extracted with ethylacetate (3.times.100 ml). The
organic layers were washed with water (1.times.) and brine
(2.times.), dried over Na.sub.2SO.sub.4, and concentrated for a
crude yellow solid, which was recrystallized from hot methanol for
0.98 g of a yellow crystalline solid (50% yield). LCMS (M+H) showed
the desired Compound V.
[0364] Part E: 186
[0365] To a solution of Compound V (0.98 g, MW 630.86, 1.0 eq) in
methylene chloride (4 ml) was added trifluoroacetic acid (4 ml,
TFA). The mixture was stirred for 4 hr at room temperature. The
mixture was then concentrated to 1/3 volume, and diethylether was
added to afford a solid, which was collected and dried for 1.0 g
tan solid (93% yield). .sup.1H NMR and LCMS showed the desired
Compound VI.
[0366] Part F: 187
[0367] To a solution of Compound VI (1.0 g, MW 688.77, 1.0 eq) in
N,N-dimethylformamide (5 ml) was added triethylamine (Aldrich, 0.8
ml, MW 101.19, 4.0 eq) followed by N-hydroxybenzotriazole hydrate
(Aldrich, 0.38 g, MW 135.13, 2.0 eq),)-(tetrahydro-2H-pyran-2-yl),
hydroxylamine (0.25 g, MW 117.16, 1.5 eq), and, lastly,
1-(3-dimethylaminopropyl)-3-ethylcarb- odiimide hydrochloride
(Sigma, 0.59 g, MW 191.76, 2.2 eq). The mixture stirred at ambient
temperature for 18 hr. To work up the reaction was diluted with
water (10 ml) and extracted with ethylacetate (3.times.75 ml). The
organics were combined and washed with a saturated sodium
bicarbonate solution (1.times.150 ml), and brine (1.times.150 ml).
The organic was then dried over Na.sub.2SO.sub.4, and concentrated
to afford a 0.9 g of a brown oil (96% yield). .sup.1H NMR and LCMS
showed the desired Compound VII.
[0368] Part G: 188
[0369] The Compound VII (0.9 g, MW 673.88, 1.0 eq) was treated with
methanol (0.5 ml) and 4 N HCl in dioxane (5 ml) for 1 hr). The
solvents were concentrated to 1/3 the volume via an N.sub.2 stream.
Diethylether was then added to the residue to afford a solid that
was collected and dried for a brown solid (0.32 g, 40% yield).
.sup.1H NMR showed the desired Compound VIII. HRMS confirmed this
observation (theo. M+H 590.2359, observed M+H 590.2364).
Example 123
Preparation of
[0370] 189
[0371] Part A. Preparation of Aryl Fluoride Ester: 190
[0372] To a solution of molecular slieves (7.5 g), ethyl
5-[(4-fluorophenyl(sulfonyl]-4-piperidinecarboxylate, monohydrate
(15 g, 42.6 mmol) in methanol (75 mL), and acetic acid (9 mL) was
added sodium cyano borohydride (7.23 g, 115 mmol). The mixture was
stirred at room temperature for 48 hr. The mixture was then
quenched with sat. NH.sub.4.sup.+Cl.sup.-, and extracted with
CH.sub.2Cl.sub.2 three times. The combined organic layer was dried
and concentrated in vacuo. The residue was then recrystallized
using ethanol and ether to give 12.1 g (32.7 mmol) of the aryl
fluoride ester.
[0373] Part B. Aryl fluoride displacement. A solution of the aryl
fluoride from Part A (0.45 mmol), Cs.sub.2CO.sub.3 (1.35 mmol, 3
eq), and 4-(4-chlorobenzoyle)piperidine (Maybridge, England, 0.67
mmol, 1.5 eq) in DMSO (1 mL) was heated to 1110.degree. C. for
18-48 h. The mixture was cooled, dissolved in saturated aqueous
NH.sub.4.sup.+Cl.sup.- (5 mL), and extracted with dichloromethane
(3.times.3 mL). The combined organic layer was blown down, and the
crude product was purified by crystallization using ethanol and
ether.
[0374] Part C. Converting the ethyl ester to the hydroxamic acid. A
solution of ethyl ester (5 g) in ethanol (8 mL), tetrahydrofuran (4
mL), and 50% aqueous NaOH (2 mL) was heated to 50.degree. C. for
approximately 2 hr (additional ethanol and THF can be added if the
solid was not completely soluble after 1 hr at 50.degree. C.). The
residue was neutralized to a pH of 5-6 with aqueous HCl. The
aqueous layer was concentrated in vacuo, and the resulting solid
was washed with acetonitrile and water, and dried under high
vacuum. A solution of the acid, NMM (3 eq), EDC (1.4 eq), and HOBT
(1.5 eq) in DMF (5 mL) was heated at 40.degree. C. for 2 hr. The
amine was added, and then stirred at room temperature for 18-48 hr.
The reaction mixture was quenched with saturated aqueous
NH.sub.4.sup.+Cl.sup.-, and extracted with dichloromethane. The
combined organic layer was concentrated. The THP amide was purified
over SiO.sub.2 using CH.sub.2Cl.sub.2/methanol/triethy- lamine (the
THP amide may alternatively be purified by reverse-phase
chromatography). The resulting solid was then dissolved in 10 mL of
4M HCl and 10 mL of methanol, and stirred at room temperature until
completion (30 min to 120 min). The mixture was then blown down,
and the resulting solid was re-dissolved in methanol and poured
into isopropyl alcohol. The solid was collected and dried. THEO
M+H=560.1986; observed HI RES M+H=560.1999.
Example 124
Preparation of
[0375] 191
[0376] Part A. Preparation of
4-(4-metholcyclopropylbenzoyl)piperidine. To a solution of
4-bromophenylcyclopropyl ketone (Acros, 20 g, 89 mmol) in THF (75
mL) was added sodium borohydride (2.25 g, 60 mmol) and aluminum
trichloride (3.95 g, 30 mmol) in small portions at -5.degree. C.
The mixture was allowed to warm to room temperature for 18 hr, and
then stirred an additional 3 hr at 40.degree. C. The mixture was
then cooled, quenched with saturated NH.sub.4.sup.+Cl.sup.-, and
extracted with CH.sub.2Cl.sub.2 three times. The combined organic
layer was dried and concentrated in vacuo. The mixture was
chromatographed over 70 g of SiO.sub.2 eluting with EtOAc:Hexane
(0:100 to 10:90) to give 14.55 g (69 mmol) of 4-methyl cyclopropyl
aryl bromide. To a cooled to 0.degree. C. solution of the 4-methyl
cyclopropyl aryl bromide (7.75 g, 36.7 mmol) in 20 mL of THF was
added magnesium (55 mmol, 3 eq), followed by dibromoethane (10 uL)
in small portions, keeping the mixture cold. The solution was
stirred for 3 hr. The weinreb amide described in Example 120 (5 g,
18.4 mmol) was added at 0.degree. C., and the mixture was stirred
at room temperature for 48 hr. The mixture was then quenched with
saturated NH.sub.4.sup.+Cl.sup.-, and extracted with
CH.sub.2Cl.sub.2 three times. The combined organic layer was dried
and concentrated in vacuo. The mixture was chromatographed over 70
g of SiO.sub.2 eluting with EtOAc/Hexane (0:100 to 30:70) to give
5.54 g (16 mmol) of the desired BOC-protected piperidine. The
BOC-protected piperidine was then dissolved in 20 mL of
CH.sub.2Cl.sub.2 and 20 mL of TFA, and stirred at room temperature
for 1 hr. The mixture was concentrated in vacuo, and the residue
was treated with 5% NaOH and water, and then extracted with
CH.sub.2Cl.sub.2 three times. The combined organic layer was dried
and concentrated in vacuo to give 3.47 g (14.3 mmol) of the
4-(4-metholcyclopropylbenzoyl)piperidine.
[0377] Part B. Aryl fluoride displacement. A solution of ethyl
4-[(4-fluorophenyl)sulfonyl]-1-(2-methoxyethyl)-4-piperidinecarboxylate
(0.45 mmol), Cs.sub.2CO.sub.3 (1.35 mmol, 3 eq), and
4-(4-metholcyclopropylbenzoyl)piperidine from Part A in DMSO (1 mL)
was heated to 110.degree. C. for 18-48 h. The mixture was cooled,
dissolved in saturated aqueous NH.sub.4.sup.+Cl.sup.- (5 mL), and
extracted with dichloromethane (3.times.3 mL). The combined organic
layer was blown down, and the crude product was purified by
crystallization using ethanol and ether.
[0378] Part C. Converting the ethyl ester to the hydroxamic acid. A
solution of ethyl ester (5 g) in ethanol (8 mL), tetrahydrofuran (4
mL), and 50% aqueous NaOH (2 mL) was heated to 50.degree. C. for
approximately 2 hr (additional ethanol and THF can be added if the
solid is not completely soluble after 1 hr at 50.degree. C.). The
residue was neutralized to a pH of 5-6 with aqueous HCl. The
aqueous layer was concentrated in vacuo, and the resulting solid
was washed with acetonitrile and water, and dried under high
vacuum. A solution of the acid, NMM (3 eq), EDC (1.4 eq), and HOBT
(1.5 eq) in DMF (5 mL) was heated at 40.degree. C. for 2 hr. The
amine was added, and then stirred at room temperature for 18-48 hr.
The reaction mixture was quenched with saturated aqueous
NH.sub.4.sup.+Cl.sup.-, and extracted with dichloromethane. The
combined organic layer was concentrated. The THP amide was purified
over SiO.sub.2 using CH.sub.2Cl.sub.2/methanol/triethy- lamine (the
THP amide may alternatively be purified by reverse-phase
chromatography). The resulting solid was then dissolved in 10 mL of
4M HCl and 10 mL of methanol, and stirred at room temperature until
completion (30 min to 120 min). The mixture was then blown down,
and the resulting solid was re-dissolved in methanol and poured
into isopropyl alcohol. The solid was collected and dried. THEO
M+H=584.2794; observed HI RES M+H=584.2795.
Examples 125-387
[0379] The following compounds were prepared in a manner similar to
that used in the preceding examples. In the tables that follow, a
generic structure is shown above the table with substituent groups
being illustrated in the table along with available mass spectral
data.
7 192 Example R K MS (ES) m/z 125 193 194 126 195 196 127 197 198
128 199 200 129 201 202 130 203 204 131 205 206 132 207 208 133 209
210 516 134 211 212 135 213 214 136 215 216 600 137 217 218 530 138
219 220 636 139 221 222 596 140 223 --H 502 141 224 225 579 142 226
227 542 143 228 229 614 144 230 231 585 145 232 233 549 146 234 --H
574 147 235 236 564 148 237 238 616 149 239 240 598 151 241 242
540.2160 152 243 244 524.225 153 245 246 544.1673 154 247 248
524.2218 155 249 250 540.2155 156 251 252 560.2387 157 253 254
555.2666 158 255 256 540.2548 159 257 258 554.2698 160 259 260
562.2378 161 261 262 541.2488 162 263 264 163 265 266 593.2131 164
267 268 578.1976 165 269 270 592.2151 166 271 272 600.1865 167 273
274 579.1984 168 275 276 543.2647 169 277 278 528.2550 170 279 280
542.2700 171 281 282 550.2390 172 283 284 529.2505 173 285 286
539.2338 174 287 288 567.2653 175 289 290 560.2249 176 291 292
544.2489 177 293 294 559.2616 178 295 296 587.2933 179 297 298
580.2504 180 299 300 546.1840 181 301 302 569.2797 182 303 304
542.2349 183 305 306 578.1288 184 307 308 607.2381 185 309 310
573.1654 186 311 312 564.1965 187 313 314 576.2552 188 315 316
556.2506 189 317 318 568.2862 190 319 320 590 191 321 322 602.2935
192 323 324 584.1298 193 325 326 534.1860 194 327 328 530.2332 195
329 330 196 331 332 580.2848 197 333 334 594.3011 198 335 336
608.3148 199 337 338 608.3152 200 339 340 582.2997 201 341 342
598.296 202 343 344 612.3124 203 345 346 626.3276 204 347 348
626.3268 205 349 350 600.3107 206 351 352 580.1822 207 353 354
546.1850 208 355 356 514.2382 210 357 358 540.2539 211 359 360
578.2106 212 361 362 558.2667 213 363 364 546.1847 214 365 366
560.2012 215 367 368 570.2608 216 369 370 584.2755 217 371 372
598.2953 218 373 374 586 219 375 376 598.1441 220 377 378 578.1966
221 379 380 548.1988 222 381 382 528.2543 223 383 384 593.2431 224
385 386 578.2359 225 387 388 564.2202 226 389 390 614.3261 227 391
392 614.2932 228 393 394 656.3399 229 395 396 230 397 398 614.3273
231 399 400 602.2901 232 401 402 568.2876 233 403 404 570.2970 234
405 406 584.3166 235 407 408 572.2787 236 409 410 584.3161 237 411
412 596.3014 238 413 414 560.2437 239 415 416 614.2883 240 417 418
629.3014 242 419 420 243 421 422 622.2636 244 423 424 616.3040 245
425 426 602.2876 246 427 428 600.3109 247 429 430 586.2949 248 431
432 572.2778 249 433 434 570.3007 250 435 436 606.2664 252 437 438
580.2147 253 439 440 587.2914 254 441 442 554.5692 256 443 444
541.1650 257 445 446 574.2388 258 447 448 543.2291 259 449 450
500.2019 260 451 452 514.2376 261 453 454 516.1723 262 455 456
518.2130 263 457 458 514.2194 264 459 460 518.2432 265 461 462
514.2375 266 463 464 530.1880 267 465 466 532.2307 268 467 468
528.2557 269 469 470 516.2557 270 471 472 518.1880 271 473 474
536.1979 272 475 476 498.2450 273 477 478 512.2615 274 479 480
532.2061
[0380]
8 481 Ex- am- ple R K MS (ES) m/z 275 482 483 578.2068 276 484 485
594.2005 277 486 487 578.2053
[0381]
9 488 Example R MS (ES) m/z 278 489 463.1704 279 490 499.2304 280
491 281 492 495.4984 282 493 479.1416 283 494 572.2800 284 495
539.2017 285 496 489.2049 286 497 477 287 498 515 288 499 483.1992
289 500 503 290 501 487 291 502 487 292 503 491 293 504 503 294 505
473 295 506 509 296 507 557 297 508 557 298 509 541 299 510 491 300
511 541 301 512 501 302 513 509 303 514 501 304 515 501 305 516 517
306 517 521 307 518 505 308 519 501 309 520 559 310 521 311 522 499
312 523 499 313 524 515 314 525 529 315 526 516 316 527 517 317 528
318 529 517 319 530 320 531 321 532 322 533 323 534 324 535 325 536
326 537 327 538 328 539 329 540 330 541 331 542 332 543 333 544 334
545 335 546 336 547 337 548 338 549 339 550 340 551 341 552 342 553
343 554 344 555 345 556 346 557 437 558 348 559 349 560 350 561 351
562 352 563 353 564 354 565 355 566 356 567 357 568 358 569 359 570
360 571 361 572 362 573 363 574 364 575 365 576 366 577 367 578 368
579 369 580 370 581 371 582 372 583 373 584 374 585 375 586 581 376
587 545.2320 377 588 529.2383 378 589 551.0854 379 590 555.252 380
591 569.2687 381 592 569.2676 382 593 543.2524 383 594 384 595
530.2315 385 596 556.2482 386 597 387 598
Example 388
In Vitro Metalloprotease Inhibition
[0382] Several hydroxamates and salts thereof were assayed for MMP
activity by an in vitro assay generally following the procedures
outlined et al., FEBS Lett., 296(3), 263 (1992).
[0383] Recombinant human MMP-1, MMP-2, MMP-9, MMP-13, and were used
in this assay. These enzymes were prepared in the Assignee's es
following usual laboratory procedures. Specifics for preparing and
e enzymes can be found in the scientific literature describing
these See, e.g., Enzyme Nomenclature (Academic Press, San Diego,
Calif., d the citations therein). See also, Frije et al., J Biol.
Chem., 26(24), (1994).
[0384] The MMP-1 was obtained from MMP-1 expressing transfected
HT-1080 cells provided by Dr. Harold Welgus of Washington
University in St. Louis, Mo. The MMP-1 was activated using
4-aminophenylmercuric acetate (APMA), and then purified over a
hydroxamic acid column.
[0385] The MMP-2 was obtained from MMP-2 expressing transfected
cells provided by Dr. Gregory Goldberg of Washington
University.
[0386] The MMP-9 was obtained from MMP-9 expressing transfected
cells provided by Dr. Gregory Goldberd.
[0387] The MMP-13 was obtained as a proenzyme from a full-length
cDNA clone using baculovirus, as described by V. A. Luckow, "Insect
Cell Expression Technology," Protein Engineering: Principles and
Practice, pp. 183-218 (edited by J. L. Cleland et al., Wiley-Liss,
Inc., 1996). The expressed proenzyme was first purified over a
heparin agarose column, and then over a chelating zinc chloride
column. The proenzyme was then activated by APMA for use in the
assay. Further details on baculovirus expression systems may be
found in, for example, Luckow et al., J. Virol., 67, 4566-79
(1993). See also, O'Reilly et al, Baculovirus Expression Vectors: A
Laboratory Manual (W.H. Freeman and Co., New York, N.Y., 1992). See
also, King et al., The Baculovirus Expression System: A Laboratory
Guide (Chapman & Hall, London, England, 1992).
[0388] The enzyme substrate was a methoxycoumarin-containing
polypeptide having the following sequence:
[0389] MCA-ProLeuGlyLeuDpaAlaArgNH.sub.2
[0390] Here, "MCA" is methoxycoumarin and "Dpa" is
3-(2,4-dinitrophenyl)-L- -2,3-diaminopropionyl alanine. This
substrate is commercially available from Baychem (Redwood City,
Calif.) as product M-1895.
[0391] The subject hydroxamate (or salt thereof) was dissolved at
various concentrations using 1% dimethyl sulfoxide (DMSO) in a
buffer containing 100 mM Tris-HCl, 100 mM NaCl, 10 mM CaCl.sub.2,
and 0.05% polyethyleneglycol (23) lauryl ether at a pH of 7.5.
These solutions were then compared to a control (which contained
equal amount of DMSO/buffer solution, but no hydroxamate compound)
using Microfluor.TM. White Plates (Dynatech, Chantilly, Va.).
Specifically, The MMPs were activated with APMA or trypsin. Then
the various hydroxamate/DMSO/buffer solutions were incubated in
separate plates at room temperature with the activated MMP and 4 um
of the MMP substrate. The control likewise was incubated at room
temperature in separate plates with the MMP and 4 uM of the MMP
substrate. In the absence of inhibitor activity, a fluorogenic
peptide was cleaved at the gly-leu peptide bond of the substrate,
separating the highly fluorogenic peptide from a 2,4-dinitrophenyl
quencher, resulting in an increase of fluorescent intensity
(excitation at 328 nm/emission at 415). Inhibition was measured as
a reduction in fluorescent intensity as a function of inhibitor
concentration using a Perkin Elmer (Norwalk, Conn.) L550 plate
reader. The IC.sub.50's were then calculated from these
measurements. The results are set forth in the following Table
A.
10 Inhibition Table A (nM) Example MMP-13 MMP-2 MMP-1 Number
IC.sub.50(nM) IC.sub.50(nM) IC.sub.50(nM) 4 15.6 2,900 >10000 5
15.6 2,900 >10000 6 18.1 >10000 >10000 7 18.0 4,500
>10000 8 50.0 2,500 >10000 9 12.2 5,600 >10000 10 40.0
6,000 >10000 11 37.0 2,700 >10000 12 6.70 1,400 >10000 13
31.6 3,500 >10000 14 45.0 >10000 >10000 15 28.0 5,500
>10000 16 42.5 4,800 >10000 17 70.0 7,000 >10000 18
>10000 >10000 >10000 19 90.0 10,000 >10000 20 23.5
4,500 >10000 21 6.00 1,600 >10000 22 10.7 3,600 >10000 23
6.40 1,600 >10000 24 6.70 700 >10000 25 4.00 445 >10000 28
10.0 800 >10000 29 20.0 4,500 >10000 30 18.1 >10000
>10000 31 15.8 2,100 >10000 32 30.0 1,750 >10000 33 67.4
6,000 67.4 34 19.3 3,700 >10000 35 26.8 900 >10000 36 70.0
5,400 >10000 37 82.5 >10000 >10000 38 17.9 5,000 >10000
39 19.0 1,050 >10000 40 80.0 5,700 >10000 41 11.4 6,000
>10000 42 20.0 6,500 >10000 44 40.0 5,700 >10000 45 10.0
>10000 >10000 46 20.0 2,000 >10000 47 4.10 562 >10000
48 0.2 0.3 3,000 49 2.00 59.0 >10000 50 50.0 5,000 >10000 51
2.20 0.45 >10000 52 32.6 900 >10000 53 27.8 7,000 >10000
58 28.8 900 >10000 59 110 1,000 >10000 60 11.4 1,200
>10000 70 43.5 2,050 >10000 72 80.0 10,000 >10000 73 9.00
8,300 >10000 74 76.9 10,000 >10000 75 4.80 >10000
>10000 76 32.7 2,700 >10000 77 160 >10000 >10000 78
70.0 >10000 >10000 79 37.3 >10000 >10000 80 70.0
>10000 >10000 81 19.3 >10000 >10000 82 20.0 7,300
>10000 83 90.0 >10000 >10000 84 105 >10000 >10000 85
14.8 9,000 >10000 86 13.8 >10000 >10000 87 130 >10000
>10000 88 19.3 9,000 >10000 89 60.0 >10000 >10000 90
150 >10000 >10000 91 35.0 >10000 >10000 92 50.0
>10000 >10000 93 50.0 >10000 >10000 95 100 >10000
>10000 96 63.1 >10000 >10000 97 59.1 >10000 >1,000
98 50.0 >10000 >10000 99 50.0 >10000 >10000 100 34.9
>10000 >10000 101 40.0 >10000 >10000 102 30.6 9,000
>10000 103 37.3 >10000 >10000 104 90.0 >10000 >10000
105 175 >10000 >10000 106 115 >10000 >10000 107 30.6
7,000 >10000 108 28.6 >10000 >10000 109 60.0 >10000
>10000 110 40.0 >10000 >10000 111 40.0 10,000 >10000
112 48.5 >10000 >10000 113 60.0 10,000 >10000 114 120
>10000 >10000 115 200 >10000 >10000 116 77.0 >10000
>10000 117 65.0 >10000 >10000 118 420 >10000 >10000
119 1.0 200 >10000 120 0.85 126 >10000 (an average of 2 (an
average of 2 experiments) experiments) 121 0.1 58.8 >10000 122
0.1 106.5 123 0.1 46.3 >10000 124 0.4 56.4 >10000 126 11.1
400 127 3.0 80.0 128 5.5 230 129 11.4 260 130 3.0 700 >10000 132
50.0 430 133 1.7 16.1 >10000 134 4.5 427 >10000 135 0.5 8.0
136 50.4 246 >10000 137 0.7 4.5 >10000 138 5.9 1500 >10000
139 1.8 330 >10000 140 18.1 800 >10000 141 1.4 160 >10000
142 6.0 420 >10000 143 2.1 100 >10000 145 210 2100 >10000
146 4.0 200 >10000 147 20.0 145 >10000 148 2.9 80.0 >10000
149 16.9 210 >10000 151 1.3 127.6 >10000 152 0.6 56.3
>10000 153 0.2 30.6 >10000 154 2.4 176.5 >10000 155 1.4
43.8 >10000 156 0.7 1335.9 >10000 157 2.7 781.6 >10000 158
2.4 217.8 >10000 159 0.5 32.2 160 0.4 197.5 >10000 161 0.3
234.7 162 2.7 494.6 >10000 163 3.4 3231.9 >10000 164 5.4
942.3 >10000 165 85.9 1754 166 438 >10000 167 4.7 2949 168
2.1 2181.2 >10000 169 2.6 1061.7 >10000 170 1.3 134.1
>10000 171 1.9 405.4 >10000 172 3.1 649.1 173 0.9 117.3 174
1.1 1069.1 >10000 175 0.7 136.6 >10000 176 0.4 122.3
>10000 177 1.4 166.8 178 3.0 1976.5 179 0.7 161.3 >10000 180
0.3 52.7 >10000 181 2.3 935.7 >10000 182 1.1 115.4 >10000
183 0.7 37.9 >10000 184 1.5 360.2 >10000 185 5.1 87.4
>10000 186 3.5 94.4 >10000 187 2.7 242.4 >10000 188 2.0
249.9 >10000 189 <0.1 258 >10000 190 0.2 23.1 >10000
191 3.0 2286.9 >10000 192 1.3 103.3 >10000 193 0.4 98.7
>10000 194 9.1 1229.7 >10000 195 0.3 462.8 >10000 196 1.0
750.1 >10000 197 1.4 1720.1 >10000 198 12.0 2565.6 199 11.7
3390.0 >10000 200 0.5 1398.8 >10000 201 0.2 6315.4 >10000
202 0.4 1017.6 >10000 203 0.6 816.4 2367 204 0.2 1045.8
>10000 205 <0.1 411.5 >10000 206 1.8 199.4 >10000 207
1.1 4.4 >10000 208 0.1 19.6 >10000 210 1.1 13.1 >10000 211
1.2 122.3 >10000 212 0.2 109.7 >10000 213 0.5 25.8 >10000
214 1.7 159.8 >10000 215 0.9 22.7 >10000 216 1.5 46.4
>10000 217 1.3 270.0 >10000 218 0.2 75.7 >10000 219 4.9
258.2 >10000 220 1.7 289.8 >10000 221 3.4 301.1 >10000 222
1.0 196.6 >10000 223 2.5 80.4 >10000 224 0.4 72.9 >10000
225 0.2 40.8 >10000 226 <0.1 1024 >10000 227 1.4 132.1
>10000 228 19.5 154.6 229 0.2 8.5 >10000 230 0.1 745.0
>10000 231 0.5 39.4 >10000 232 1.3 624.4 >10000 233 1.2
1046.1 >10000 234 7.5 2444.7 >10000 235 0.8 118.0 >10000
236 1.5 1848.4 >10000 237 2.1 1914.8 >10000 238 1.8 62.1 239
0.6 75.8 240 2.8 86.0 242 1.0 87.5 243 0.3 56.0 >10000 244 0.2
15.2 245 1.1 38.6 246 1.0 2712.9 >10000 247 0.3 111.4 >10000
248 0.6 141.0 >10000 249 5.8 >10000 >10000 250 2.1 107.2
252 0.4 14.3 253 1.7 38.7 >10000 254 1.3 132.0 >10000 256 7.5
35.4 257 258 14.1 45.4 259 0.4 0.6 260 0.4 1.2 >10000 261 0.8
1.0 262 1.0 1.7 263 1.5 2.6 >10000 264 0.8 3.1 265 0.5 3.2 266
1.7 4.5 267 0.4 1.7 >10000 268 1.2 5.0 >10000 269 1.4 4.5
>10000 270 1.1 1.9 >10000 271 0.8 1.7 >10000 272 1.3 5.9
>10000 273 2.5 13.4 274 2.1 5.2 >10000 275 183.6 6736.9 276
126.7 2733.4 277 274.5 >10000 279 160 3300 >10000 280 27.1
500 >10000 281 11.4 500 >10000 282 0.7 2.0 >10000 284 33.7
5400 >10000 285 35.0 3100 >10000 287 70.0 >10000 >10000
288 4.4 60.7 >10000 289 6.0 160 >10000 290 0.4 82.0 >10000
291 0.8 160 >10000 292 3.2 35.0 >10000 293 37.3 1400
>10000 294 3.1 120 >10000 295 28.6 300 >10000 296 25.1 210
>10000 297 15.8 250 >10000 298 34.9 240 >10000 299 9.4 106
>10000 300 14.8 240 >10000 301 37 3000 >10000 302 1.9 35
>10000 303 3.1 590 >10000 304 1.6 270 >10000 305 6.0 3300
>10000 306 9.0 800 >10000 307 0.9 145 >10000 308 3.0 1280
>10000 309 22.0 270 >10000 310 6.0 4500 >10000 311 3.7 700
>10000 312 1.2 175 >10000 313 3.0 445 >10000 314 12.2 3700
>10000 315 4.5 700 >10000 316 2.0 700 >10000 317 4.0 23.5
>10000 318 5.7 130 >10000 319 4.0 175 >10000 320 2.3
10,000 >10000 321 200 1400 >10000 322 140 1400 >10000 323
7.0 505 >10000 324 11.3 70.0 >10000 325 11.0 1750 >10000
326 3.0 70.0 >10000 327 5.0 4700 >10000 328 4.5 186 >10000
329 20.0 1800 ND 330 -- -- ND 331 1.2 250 ND 332 1.3 120 ND 333 3.7
600 >10000 334 5.5 440 ND 335 2.7 1500 >10000 336 2.0 34.9 ND
337 1.7 40.0 ND 338 -- -- ND 339 -- -- ND 340 16.5 10,000 >10000
341 -- -- ND 342 2.0 76.9 ND 374 5.6 970 >10000 375 34.4 2663
376 6.4 2185.4 >10000 377 0.4 361.4 >10000 378 0.3 28.4
>10000 379 0.6 1266.4 >10000 380 9.7 2287.6 >10000 381 3.5
639.9 >10000 382 0.3 1305.4 >10000 383 36.9 382.4 384 2.9
52.9 385 3.2 34.6 386 15.2 1901.1 387 4.5 344.4
Example 389
In Vivo Angiogenesis Assay
[0392] The study of angiogenesis depends on a reliable and
reproducible model for the stimulation and inhibition of a
neovascular response. The corneal micropocket assay provides such a
model of angiogenesis in the cornea of a mouse. See, A Model of
Angiogenesis in the Mouse Cornea; Kenyon, B M, et al.,
Investigative Ophthalmology & Visual Science, July 1996, Vol.
37, No. 8.
[0393] In this assay, uniformly sized Hydron.TM. pellets containing
bFGF and sucralfate were prepared and surgically implanted into the
stroma mouse cornea adjacent to the temporal limbus. The pellets
were formed by making a suspension of 20 .mu.L sterile saline
containing 10 .mu.g recombinant bFGF, 10 mg of sucralfate and 10
.mu.L of 12 percent Hydron.TM. in ethanol. The slurry was then
deposited on a 10.times.10 mm piece of sterile nylon mesh. After
drying, the nylon fibers of the mesh were separated to release the
pellets.
[0394] The corneal pocket is made by anesthetizing a 7 week old
C57Bl/6 female mouse, then proptosing the eye with ajeweler's
forceps. Using a dissecting microscope, a central, intrastromal
linear keratotomy of approximately 0.6 mm in length is performed
with a #15 surgical blade, parallel to the insertion of the lateral
rectus muscle. Using a modified cataract knife, a lamellar
micropocket is dissected toward the temporal limbus. The pocket is
extended to within 1.0 mm of the temporal limbus. A single pellet
was placed on the corneal surface at the base of the pocket with a
jeweler's forceps. The pellet was then advanced to the temporal end
of the pocket. Antibiotic ointment was then applied to the eye.
[0395] Mice were dosed on a daily basis for the duration of the
assay. Dosing of the animals was based on bioavailability and
overall potency of the compound. an exemplary dose was 10 or 50
mg/kg (mpk) bid, po. Neovascularization of the corneal stroma
begins at about day three and was permitted to continue under the
influence of the assayed compound until day five. At day five, the
degree of angiogenic inhibition was scored by viewing the
neovascular progression with a slit lamp microscope.
[0396] The mice were anesthetized and the studied eye was once
again proptosed. The maximum vessel length of neovascularization,
extending from the limbal vascular plexus toward the pellet was
measured. In addition, the contiguous circumferential zone of
neovascularization was measured as clock hours, where 30 degrees of
arc equals one clock hour. The area of angiogenesis was calculated
as follows. 1 area = ( 0.4 .times. clock hours .times. 3.14 .times.
vessel length ( in mm ) ) 2
[0397] Five to six mice were utilized for each compound in each
study. The studied mice were thereafter compared to control mice
and the difference in the area of neovascularization was recorded
as an averaged value. Each group of mice so studied constitutes an
"n" value of one, so that "n" values greater than one represent
multiple studies whose averaged result is provided in the table. A
contemplated compound typically exhibits about 25 to about 75
percent inhibition, whereas the vehicle control exhibits zero
percent inhibition.
Example 390
In Vivo PC-3 Tumor Reduction
[0398] PC-3 human pancreatic cancer eclls (ATCC CRL 1435) were
grown to 90% confluence in F12/MEM (Gibco) containing 7% FBS
(Gibco). Cells were mechanically harvested using a rubber scraper,
and then washed twice with cold medium. The resulting cells were
resuspended in cold medium with 30% matrigel (Collaborative
Research) and the cell-containing medium was maintained on ice
until used.
[0399] Balb/c nu/nu mice at 7-9 weeks of age were anesthetized with
avertin [2,2,2-tribromethanol/t-amyl alcohol (1 g/1 mL) diluted
1:60 into phosphate-buffered sline] and 3-5.times.10.sup.6 of the
above cells in 0.2 mL of medium were injected into the left flank
of each mouse. Cells were injected in the morning, whereas dosing
with an inhibitor began at 6 PM. The animals were gavaged BID from
day zero (cell injection day) to day 25-30, at which time the
animals were euthanized and tumors weighed.
[0400] Compounds were dosed at 10 mg/mL in 0.5%
methylcellulose/0.1% polysorbate 80 to provide a 50 mg/kg (mpk)
dose twice each day, or diluted to provide a 10 mg/kg (mpk) dose
twice each day. Tumor measurements began on day 7 and continued
every third or fourth day until completion of the study. Groups of
ten mice were used in each study and nine to ten survived. Each
group of mice so studied constitutes an "n" value of one, so that
"n" values greater than one represent multiple studies whose
averaged result is provided in the table.
Example 391
Tumor Necrosis Factor Assays
[0401] Cell Culture.
[0402] The cells used in the assay are the human moncytic line
U-937 (ATCC CRL-1593). The cells are grown in RPMI w/10% FCS and
PSG supplement (R-10) and are not permitted to overgrow. The assay
is carried out as follows:
[0403] 1. Count, then harvest cells by centrifugation. Resuspend
the pellet in R-10 supplement to a concentration of
1.540.times.10.sup.6 cells/mL.
[0404] 2. Add test compound in 65 uL R-10 to the appropriate wells
of a 96-well flat bottom tissue culture plate. The initial dilution
from a DMSO stock (100 mM compound) provides a 400 uM solution,
from which five additional three-fold serial dilutions are made.
Each dilution of 65 ul (in triplicate) yields final compound test
concentrations of 100 .mu.M, 33.3 .mu.M, 11.1 .mu.M, 3.7 .mu.M, 1.2
.mu.M and 0.4 .mu.M.
[0405] 3. The counted, washed and resuspended cells (200,000
cells/well) in 130 .mu.L are added to the wells.
[0406] 4. Incubation is for 45 min to 1 hr at 37.degree. C. in 5%
CO.sub.2 in a water saturated container.
[0407] 5. R-10 (65 uL) containing 160 ng/mL PMA (Sigma) is added to
each well.
[0408] 6. The test system is incubated at 37.degree. C. in 5%
CO.sub.2 overnight (18-20 hr) under 100% humidity.
[0409] 7. Supernatant, 150 .mu.L, is carefully removed from each
well for use in the ELISA assay.
[0410] 8. For toxicity, a 50 .mu.L aliquot of working solution
containg 5 mL R-10, 5 mL MTS solution [CellTiter 96 AQueous One
Solution Cell Proliferation Assay Cat.#G358/0,1 (Promega Biotech)]
and 250 ul PMS solution are added to each well containing the
remaining supernatant and cells and the cells incubated at
37.degree. C. in 5% CO.sub.2 until the color develops. The system
is excited at 570 nm and read at 630 nm.
[0411] TNF Receptor II ELISA Assay
[0412] 1. Plate 100 .mu.L/well 2 .mu.g/mL mouse anti-human TNFrII
antibody (R&D Systems #MAB226) in 1.times.PBS (pH 7.1, Gibco)
on NUNC-Immuno Maxisorb plate. Incubate the plate at 4.degree. C.
overnight (about 18-20 hr).
[0413] 2. Wash the plate with PBS-Tween (1.times.PBS w/0.05%
Tween).
[0414] 3. Add 200 .mu.L 5% BSA in PBS and block at 37.degree. C. in
a water saturated atmosphere for 2 hr.
[0415] 4. Wash the plate with PBS-Tween.
[0416] 5. Add sample and controls (100 ul of each) to each well.
The standards are 0, 50, 100, 200, 300 and 500 pg recombinant human
TNFrII (R&D Systems #226-B2) in 100 .mu.L 0.5% BSA in PBS. The
assay is linear to between 400-500 pg of standard.
[0417] 6. Incubate at 37.degree. C. in a saturated atmosphere for
1.5 hr.
[0418] 7. Wash the plate with PBS-Tween.
[0419] 8. Add 100 .mu.L goat anti-human TNFrII polyclonal (1.5
.mu.g/mL R&D Systems #AB226-PB in 0.5% BSA in PBS).
[0420] Incubate at 37.degree. C. in a saturated atmosphere for 1
hr.
[0421] 10. Wash the plate with PBS-Tween.
[0422] 11. Add 100 .mu.L anti-goat IgG-peroxidase (1:50,000 in 0.5%
BSA in PBS, Sigma #A5420).
[0423] 11. Incubate at 37.degree. C. in a saturated atmosphere for
1 hr.
[0424] 12. Wash the plate with PBS-Tween.
[0425] 13. Add 10 .mu.L KPL TMB developer, develop at room
temperature (usually about 10 min); then terminate with phosphoric
acid and excite at 450 nm and read at 570 nm.
[0426] TNF.alpha. ELISA Assay.
[0427] Coat Immulon.RTM. 2 plates with 0.1 mL/well of lug/mL
Genzyme mAb in 0.1 M NaHCO3 pH 8.0 buffer overnight (about 18-20
hr) at 4.degree. C., wrapped tightly in Saran.RTM. wrap.
[0428] Flick out coating solution and block plates with 0.3 mL/well
blocking buffer overnight at 4.degree. C., wrapped in Saran.RTM.
wrap.
[0429] Wash wells thoroughly 4.times. with wash buffer and
completely remove all wash buffer. Add 0.1 mL/well of either
samples or rhTNF.alpha. standards. Dilute samples if necessary in
appropriate diluant (e.g. tissue culture medium). Dilute standard
in same diluant. Standards and samples should be in
triplicates.
[0430] Incubate at 37.degree. C. for 1 hr in humified
container.
[0431] Wash plates as above. Add 0.1 mL/well of 1:200 dilution of
Genzyme rabbit anti-hTNFa.
[0432] Repeat incubation.
[0433] Repeat wash. Add 0.1 mL/well of 1 .mu.g/mL Jackson goat
anti-rabbit IgG (H+L)-peroxidase.
[0434] Incubate at 37.degree. C. for 30 min.
[0435] Repeat wash. Add 0.1 mL/well of peroxide-ABTS solution.
[0436] Incubate at room temperature for 5-20 min.
[0437] Read OD at 405 nm.
[0438] 12 Reagents are:
[0439] Genzyme mouse anti-human TNF? monoclonal (Cat.#
80-3399-01)
[0440] Genzyme rabbit anti-human TNF? polyclonal (Cat.#IP-300)
[0441] Genzyme recombinant human TNF? (Cat.#TNF-H).
[0442] Jackson Immunoresearch peroxide-conjugated goat anti-rabbit
IgG (H+L) (Cat.#111-035-144).
[0443] Kirkegaard/Perry peroxide ABTS solution (Cat#50-66-01).
[0444] Immulon 2 96-well microtiter plates.
[0445] Blocking solution is 1 mg/mL gelatin in PBS with 1.times.
thimerasol.
[0446] Wash buffer is 0.5 mL Tween.RTM. 20 in 1 liter of PBS.
Example 392
In Vitro Aggrecanase Inhibition Assay
[0447] Assays for measuring the potency (IC.sub.50) of a compound
toward inhibiting aggrecanase are known in the art.
[0448] One such assay, for example, has been reported in European
Patent Application Publ. No. EP 1 081 137 A1. In that assay,
primary porcine chondrocytes from articular joint cartilage are
isolated by sequential trypsin and collagenase digestion followed
by collagenase digestion overnight and are plated at
2.times.10.sup.5 cells per well into 48 well plates with 5
.mu.Ci/ml.sup.35S (1000 Ci/mmol) sulphur in type 1 collagen coated
plates. Cells are allowed to incorporate label into their
proteoglycan matrix (approximately 1 week) at 37.degree. C. under
an atmosphere of 5% CO.sub.2. The night before initiating the
assay, chondrocyte monolayers are washed 2 times in DMEM/1% PSF/G
and then allowed to incubate in fresh DMEM/1% FBS overnight. The
next morning, chondrocytes are washed once in DMEM/1% PSF/G. The
final wash is allowed to sit on the plates in the incubator while
making dilutions. Media and dilutions are made as described in the
following Table C:
11TABLE C control media DMEM alone IL-1 media DMEM + IL-1 (5 ng/ml)
drug dilutions Make all compound stocks at 10 mM in DMSO. Make a
100 .mu.M stock of each compound in DMEM in 96-well plate. Store in
freezer overnight. The next day, perform serial dilutions in DMEM
with IL-1 to 5 .mu.M, 500 nM, and 50 nM. Aspirate final wash from
wells and add 50 .mu.M of compound from above dilutions to 450
.mu.L of IL-1 media in appropriate wells of the 48 well plates.
Final compound concentrations equal 500 nM, 50 nM, and 5 nM. All
samples completed in triplicate with control and IL-1 alone on each
plate.
[0449] Plates are labeled and only the interior 24 wells of the
plate are used. On one of the plates, several columns are
designated as IL-1 (no drug) and control (no IL-1, no drug). These
control columns are periodically counted to monitor
35S-proteoglycan release. Control and IL-1 media are added to wells
(450/L) followed by compound (50 .mu.L) so as to initiate the
assay. Plates are incubated at 37.degree. C. with 5% CO.sub.2
atmosphere. At 40-50% release (when CPM from IL-1 media is 4-5
times control media) as assessed by liquid scintillation counting
(LSC) of media samples, the assay is terminated (about 9 to about
12 hours). Media is removed from all wells and placed into
scintillation tubes. Scintillate is added and radioactive counts
are acquired (LSC). To solubilize cell layers, 500 .mu.L of papain
digestion buffer (0.2 M Tris, pH 7.0, 5 mM DTT, and 1 mg/ml papain)
is added to each well. Plates with digestion solution are incubated
at 60.degree. C. overnight. The cell layer is removed from the
plates the next day and placed in scintillation tubes. Scintillate
is then added, and samples counted (LSC). The percent of released
counts from the total present in each well is determined. Averages
of the triplicates are made with control background subtracted from
each well. The percent of compound inhibition is based on IL-1
samples as 0% inhibition (100% of total counts).
[0450] Another assay for measuring aggrecanase inhibition has been
reported in WIPO Int'l Publ. No. WO 00/59874. That assay reportedly
uses active aggrecanase accumulated in media from stimulated bovine
cartilage (BNC) or related cartilage sources and purified cartilage
aggrecan monomer or a fragment thereof as a substrate. Aggrecanase
is generated by stimulation of cartilage slices with interleukin-1
(IL-1), tumor necrosis factor alpha (TNF-.alpha.), or other
stimuli. To accumulate BNC aggrecanase in culture media, cartilage
reportedly is first depleted of endogenous aggrecan by stimulation
with 500 ng/ml human recombinant IL-.beta. for 6 days with media
changes every 2 days. Cartilage is then stimulated for an
additional 8 days without media change to allow accumulation of
soluble, active aggrecanase in the culture media. To decrease the
amounts of matrix metalloproteinases released into the media during
aggrecanase accumulation, agents which inhibit MMP-1, -2, -3, and
-9 biosynthesis are included during stimulation. This BNC
conditioned media containing aggrecanase activity is then used as
the source of aggrecanase for the assay. Aggrecanase enzymatic
activity is detected by monitoring production of aggrecan fragments
produced exclusively by cleavage at the Glu373-Ala374 bond within
the aggrecan core protein by Western analysis using the monoclonal
antibody, BC-3 (Hughes, et al., Biochem J, 306:799-804 (1995)).
This antibody reportedly recognizes aggrecan fragments with the
N-terminus, 374ARGSVIL, generated upon cleavage by aggrecanase. The
BC-3 antibody reportedly recognizes this neoepitope only when it is
at the N-terminus and not when it is present internally within
aggrecan fragments or within the aggrecan protein core. Only
products produced upon cleavage by aggrecanase reportedly are
detected. Kinetic studies using this assay reportedly yield a Km of
1.5+/-0.35 .mu.M for aggrecanase. To evaluate inhibition of
aggrecanase, compounds are prepared as 10 mM stocks in DMSO, water,
or other solvents and diluted to appropriate concentrations in
water. Drug (50/L) is added to 50 .mu.L of aggrecanase-containing
media and 50 .mu.L of 2 mg/ml aggrecan substrate and brought to a
final volume of 200 mL in 0.2 M Tris, pH 7.6, containing 0.4 M NaCl
and 40 mM CaCl.sub.2. The assay is run for 4 hr at 37.degree. C.,
quenched with 20 mM EDTA, and analyzed for aggrecanase-generated
products. A sample containing enzyme and substrate without drug is
included as a positive control and enzyme incubated in the absence
of substrate serves as a measure of background. Removal of the
glycosaminoglycan side chains from aggrecan reportedly is necessary
for the BC-3 antibody to recognize the ARGSVIL epitope on the core
protein. Therefore, for analysis of aggrecan fragments generated by
cleavage at the Glu373-Ala374 site, proteoglycans and proteoglycan
fragments are enzymatically deglycosylated with chondroitinase ABC
(0.1 units/110 g GAG) for 2 hr at 37.degree. C. and then with
keratanase (0.1 units/110 g GAG) and keratanase II (0.002 units/10
g GAG) for 2 hr at 37.degree. C. in buffer containing 50 mM sodium
acetate, 0.1 M Tris/HCl, pH 6.5. After digestion, aggrecan in the
samples is precipitated with 5 volumes of acetone and resuspended
in 30 .mu.L of Tris glycine SDS sample buffer (Novex) containing
2.5% beta mercaptoethanol. Samples are loaded and then separated by
SDS-PAGE under reducing conditions with 4-12% gradient gels,
transferred to nitrocellulose and immunolocated with 1:500 dilution
of antibody BC3. Subsequently, membranes are incubated with a
1:5000 dilution of goat anti-mouse IgG alkaline phosphatase second
antibody and aggrecan catabolites visualized by incubation with
appropriate substrate for 10-30 minutes to achieve optimal color
development. Blots are quantitated by scanning densitometry and
inhibition of aggrecanase determined by comparing the amount of
product produced in the presence versus absence of compound.
[0451] The above detailed description of preferred embodiments is
intended only to acquaint others skilled in the art with the
invention, its principles, and its practical application so that
others skilled in the art may adapt and apply the invention in its
numerous forms, as they may be best suited to the requirements of a
particular use. This invention, therefore, is not limited to the
above embodiments, and may be variously modified.
* * * * *