U.S. patent application number 09/964114 was filed with the patent office on 2002-10-24 for sulfonamide interleukin-1beta converting enzyme inhibitors.
This patent application is currently assigned to Warner-Lambert Company. Invention is credited to Albrecht, Hans P., Allen, Hamish John, Brady, Kenneth Dale, Harter, William Glen, Kostlan, Catherine Rose, Roth, Bruce David, Walker, Nigel.
Application Number | 20020156094 09/964114 |
Document ID | / |
Family ID | 21842750 |
Filed Date | 2002-10-24 |
United States Patent
Application |
20020156094 |
Kind Code |
A1 |
Albrecht, Hans P. ; et
al. |
October 24, 2002 |
Sulfonamide interleukin-1beta converting enzyme inhibitors
Abstract
The present invention relates to compounds that are inhibitors
of interleukin-1.beta. converting enzyme that have the Formula (I).
This invention also relates to a method of treatment of stroke,
reperfusion injury, Alzheimer's disease, shigellosis, inflammatory
diseases, and septic shock and to a pharmaceutically acceptable
composition that contains a compound that is an inhibitor of
interleukin-1.beta. converting enzyme.
Inventors: |
Albrecht, Hans P.;
(Gorxheimertal, DE) ; Allen, Hamish John;
(Boylston, MA) ; Brady, Kenneth Dale; (Worcester,
MA) ; Harter, William Glen; (Chelsea, MI) ;
Kostlan, Catherine Rose; (Saline, MI) ; Roth, Bruce
David; (Plymouth, MI) ; Walker, Nigel;
(Dossenheim, DE) |
Correspondence
Address: |
LAHIVE & COCKFIELD
28 STATE STREET
BOSTON
MA
02109
US
|
Assignee: |
Warner-Lambert Company
|
Family ID: |
21842750 |
Appl. No.: |
09/964114 |
Filed: |
September 25, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09964114 |
Sep 25, 2001 |
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09284422 |
Apr 9, 1999 |
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6316415 |
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09284422 |
Apr 9, 1999 |
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PCT/US97/18396 |
Oct 9, 1997 |
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60028313 |
Oct 11, 1996 |
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Current U.S.
Class: |
514/307 ;
514/601; 546/141; 564/80; 564/82 |
Current CPC
Class: |
C07K 5/1019 20130101;
C07K 5/1021 20130101; A61P 19/02 20180101; C07K 5/0812 20130101;
A61P 29/00 20180101; A61P 31/00 20180101; A61K 38/00 20130101; C07K
5/06078 20130101; C07K 5/06086 20130101; C07C 2601/14 20170501;
C07K 5/1024 20130101; A61P 9/00 20180101; C07K 5/06139 20130101;
C07K 5/1016 20130101; A61P 43/00 20180101; C07C 2602/08 20170501;
C07D 217/24 20130101; C07K 5/06104 20130101; C07K 5/0821 20130101;
C07C 311/06 20130101; C07C 311/19 20130101; C07K 5/0606 20130101;
C07K 5/1005 20130101; A61P 25/28 20180101; C07K 5/0804 20130101;
C07K 5/0819 20130101; C07C 2602/42 20170501; C07K 5/06026 20130101;
C07K 5/0815 20130101; C07C 2603/18 20170501 |
Class at
Publication: |
514/307 ;
514/601; 546/141; 564/82; 564/80 |
International
Class: |
C07D 217/22; C07C
311/30; A61K 031/47; A61K 031/18 |
Claims
1. A compound of Formula I 19R.sup.3 is hydrogen, C.sub.1-C.sub.6
alkyl, --(CH.sub.2).sub.n aryl, or --(CH.sub.2).sub.n heteroaryl;
R.sup.4 is C.sub.1-C.sub.6 alkyl, --(CH.sub.2).sub.n aryl, or
--(CH.sub.2).sub.n heteroaryl; R.sup.5 and R.sup.6 are each
independently hydrogen, C.sub.1-C.sub.6 alkyl, --(CH.sub.2).sub.n
aryl, or --(CH.sub.2).sub.n heteroaryl; R.sup.7 is C.sub.1-C.sub.6
alkyl, --(CH.sub.2).sub.n aryl, or --(CH.sub.2).sub.n heteroaryl;
each n is independently 0 to 6; each m is independently 0, 1, 2, or
3; A is alanine, leucine, isoleucine, proline, phenylalanine,
glycine, tyrosine, serine, threonine, tryptophan, cysteine,
methionine, valine, asparagine, glutamine, aspartic acid, lysine,
glutamic acid, arginine, or histidine; each R.sup.Q is
independently hydrogen or C.sub.1-C.sub.6 alkyl; R.sup.2 is
--(CH.sub.2).sub.n--Z; and Z is aryl, heteroaryl, cycloalkyl,
C.sub.1-C.sub.6alkyl, 20 fluorenyl, substituted fluorenyl,
substituted aryl, substituted heteroaryl, or substituted
cycloalkyl, and the pharmaceutically acceptable salts, esters,
amides, and prodrugs thereof.
2. A compound according to claim 1 wherein R.sup.1 is 21
3. A compound according to claim 1 wherein R.sup.1 is 22m is 0, and
R.sup.7 is --(CH.sub.2).sub.n aryl.
4. A compound according to claim 1 wherein R.sup.1 is 23m is 0, and
R.sup.7 is --CH.sub.2 aryl.
5. A compound according to claim 1 wherein R.sup.2 is
--(CH.sub.2).sub.n aryl.
6. A compound according to claim 5 wherein aryl is phenyl or
naphthyl.
7. A compound according to claim 1 wherein R.sup.2is
--(CH.sub.2).sub.n -cycloalkyl.
8. A compound according to claim 1 wherein R.sup.1 24or
--SO.sub.2-phenyl.
9. A compound according to claim 1 wherein R.sup.2 is 25
10. A compound according to claim 1 wherein R.sup.2 is 26
11. A compound of the Formula I 27wherein R.sup.2 is
--CH.sub.2CH.sub.2-- aryl, --CH.sub.2-- cycloalkyl,
--CH.sub.2CH.sub.2-- cycloalkyl, or --CH.sub.2CH.sub.2--
heteroaryl; R.sup.1 is 28R.sup.a is --(CH.sub.2).sub.n aryl or
--(CH.sub.2).sub.n heteroaryl; R.sup.b is aryl or heteroaryl;
R.sup.c is --CH.sub.2 aryl or aryl; R.sup.d is hydrogen or
C.sub.1-C.sub.6 alkyl; R.sup.e is --CH.sub.2 aryl or --CH.sub.2
heteroaryl; and the pharmaceutically acceptable salts, esters,
amides, and prodrugs thereof.
12. A compound according to claim 11 wherein R.sup.1 is 29
13. A compound according to claim 11 wherein R.sup.1 is 30
14. A compound according to claim 11 wherein R.sup.e is
--(CH.sub.2).sub.n aryl.
15. A compound according to claim 14 wherein aryl is phenyl or
naphthyl.
16. A compound according to claim 13 wherein R.sup.b is aryl.
17. A compound according to claim 16 wherein is aryl is phenyl.
18. The compounds:
3-Benzyloxycarbonylamino-4-oxo-5-(2-phenylmethanesulfon-
ylamino)-pentanoic acid;
3-Benzyloxycarbonylamino-4-oxo-5-(3-phenyl-propan-
e-1-sulfonylamino)-pentanoic acid;
3-Benzyloxycarbonylamino-4-oxo-5-phenyl-
methanesulfonyl-amino-pentanoic acid;
5-Benzenesulfonylamino-3-Benzyloxyca- rbonylamino-4-oxo-pentanoic
acid; 3-Benzyloxycarbonylarnino-5-methanesulfo-
nylamino-4-oxo-pentanoic acid;
3-Benzyloxycarbonylamino-5-(naphthalene-1-s-
ulfonylamino)-4-oxo-pentanoic acid;
3-Benzyloxycarbonylamino-5-(2-cyclohex-
yl-ethanesulfonylamino)-4-oxo-pentanoic acid;
3-Benzyloxycarbonylamino-5-(-
2-naphthalen-1-yl-ethanesulfonylamino)-4-oxo-pentanoic acid;
3-Benzyloxycarbonylamino-5-(7,7-dimethyl-2-oxo-bicyclo
[2.2.1]hept-1-(R)-ylmethanesulfonylamino)-4-oxo-pentanoic acid;
3-Benzyloxycarbonylamino-5-(indan-1-ylmethanesulfonylamino)-4-oxo-pentano-
ic acid;
3-Benzyloxycarbonylamino-5-(9-fluoro-9H-fluoren-9-ylmethanesulfon-
ylamino)-4-oxo-pentanoic acid;
3-Benzyloxycarbonylamino-5-(7,7-dimethyl-2-- oxo-bicyclo
[2.2.1]hept-1-(S)-ylmethanesulfonylamino)-4-oxo-pentanoic acid;
3-(2-Acetylamino-3-methyl-butyrylamino)-5-(7,7-dimethyl-2-oxo-bicyc-
lo[2.2.1]hept-1-(S)-ylmethanesulfonylamino)-4-oxo-pentanoic acid;
3-(2-Acetylamino-propylamino)-5-(7,7-dimethyl-2-oxo-bicyclo[2.2.1]hept-1--
(S)-ylmethanesulfonylamino)-4-oxo-pentanoic acid;
3-(1,2,3,4-tetrahydro-1--
oxo-isoquinoline-2-yl)-acetanino-5-benzenesulfonylamino-4-oxo-pentanoic
acid;
(S)-5-(Bicyclo[2.2.1]hept-1-ylmethanesulfonylamino)-4-oxo-3-[2-(1-o-
xo-3,4-dihydro-1 H-isoquinolin-2-yl)-acetylamino]-pentanoic acid;
(S)-4-Oxo-3-[2-(1-oxo-3,4-dihydro-1H-isoquinolin-2-yl)-acetylamino
]-5-(2-phenyl-ethanesulfonylamino)-pentanoic acid; and
4-Oxo-3-[2-(1-oxo-3,4-dihydro-1H-isoquinolin-2-yl)-acetylamino]-5-phenylm-
ethanesulfonylamino-pentanoic acid.
19. A method of inhibiting interleukin-1.beta. converting enzyme,
the method comprising administering to a patient in need of
inhibition of interleukin-1.beta. converting enzyme a
therapeutically effective amount of a compound of claim 1.
20. A method of inhibiting Caspase-4, the method comprising
administering to a patient in need of Caspase-4 inhibition a
Caspase-4 inhibiting amount of a compound of claim 1.
21. A method of treating or preventing stroke, the method
comprising administering to a patient having a stroke or having had
a stroke a therapeutically effective amount of a compound of claim
1.
22. A method of treating inflammatory diseases, the method
comprising administering to a patient having an inflammatory
disease a therapeutically effective amount of a compound of claim
1.
23. The method of claim 22 wherein the inflammatory disease is
arthritis.
24. The method of claim 22 wherein the inflammatory disease
inflammatory bowel disease.
25. A pharmaceutically acceptable composition that contains a
compound of claim 1.
26. A method of inhibiting interleukin-1.beta. converting enzyme,
the method comprising administering to a patient in need of
inhibition of interleukin-1.beta. converting enzyme a
therapeutically effective amount of a compound of claim 11.
27. A method of inhibiting Caspase-4, the method comprising
administering to a patient in need of Caspase-4 inhibition a
Caspase-4 inhibiting amount of a compound of claim 11.
28. A method of treating or preventing stroke, the method
comprising administering to a patient having a stroke or having had
a stroke a therapeutically effective amount of a compound of claim
11.
29. A method of treating inflammatory diseases, the method
comprising administering to a patient having an inflammatory
disease a therapeutically effective amount of a compound of claim
11.
30. The method of claim 29 wherein the inflammatory disease is
arthritis.
31. The method of claim 29 wherein the inflammatory disease is
inflammatory bowel disease.
32. A pharmaceutically acceptable composition that contains a
compound of claim 11.
33. A method of treating septic shock, the method comprising
administering to a patient having septic shock a therapeutically
effective amount of a compound of claim 1.
34. A method of treating septic shock, the method comprising
administering to a patient having septic shock a therapeutically
effective amount of a compound of claim 11.
35. A method of treating reperfusion injury, the method of
comprising administering to a patient having reperfusion injury a
therapeutically effective amount of a compound of claim 1.
36. A method of treating reperfusion injury, the method of
comprising administering to a patient having reperfusion injury a
therapeutically effective amount of a compound of claim 11.
37. A method of treating Alzheimer's disease, the method comprising
administering to a patient having Alzheimer's disease a
therapeutically effective amount of a compound of claim 1.
38. A method of treating Alzheimer's disease, the method comprising
administering to a patient having Alzheimer's disease a
therapeutically effective amount of a compound of claim 11.
39. A method of treating shigellosis, the method comprising
administering to a patient having shigellosis a therapeutically
effective amount of a compound of claim 1.
40. A method of treating shigellosis, the method comprising
administering to a patient having shigellosis a therapeutically
effective amount of a compound of claim 11.
41. A compound of the Formula II 31R.sup.a is --(CH.sub.2).sub.n--
aryl or --(CH.sub.2).sub.n heteroaryl; R.sup.b is aryl or
heteroaryl; R.sup.c is --CH.sub.2 aryl or aryl; R.sup.d is hydrogen
or C.sub.1-C.sub.6 alkyl; R.sup.e is --CH.sub.2 aryl or --CH.sub.2
heteroaryl; and the pharmaceutically acceptable salts, esters,
amides, and prodrugs thereof.
42. A compound according to claim 41 wherein R.sup.1 is 32
43. A compound according to claim 41 wherein R.sup.1 is 33
44. A compound according to claim 41 wherein R.sup.e is
--(CH.sub.2).sub.n aryl.
45. A compound according to claim 41 wherein aryl is phenyl or
naphthyl.
46. A compound according to claim 41 wherein R.sup.b is aryl.
47. A compound according to claim 46 wherein is aryl is phenyl.
48. A method of inhibiting interleukin-1.beta. converting enzyme,
the method comprising administering to a patient in need of
inhibition of interleukin-1.beta. converting enzyme a
therapeutically effective amount of a compound of claim 41.
49. A method of inhibiting Caspase-4, the method comprising
administering to a patient in need of Caspase-4 inhibition a
Caspase-4 inhibiting amount of a compound of claim 41.
50. A method of treating or preventing stroke, the method
comprising administering to a patient having a stroke or having had
a stroke a therapeutically effective amount of a compound of claim
41.
51. A method of treating inflammatory diseases, the method
comprising administering to a patient having an inflammatory
disease a therapeutically effective amount of a compound of claim
41.
52. The method of claim 51 wherein the inflammatory disease is
arthritis.
53. The method of claim 51 wherein the inflammatory disease
inflammatory bowel disease.
54. A method of treating septic shock, the method comprising
administering to a patient having septic shock a therapeutically
effective amount of a compound of claim 41.
55. A method of treating reperfusion injury, the method of
comprising administering to a patient having reperfusion injury a
therapeutically effective amount of a compound of claim 41.
56. A method of treating Alzheimer's disease, the method comprising
administering to a patient having Alzheimer's disease a
therapeutically effective amount of a compound of claim 41.
57. A method of treating shigellosis, the method comprising
administering to a patient having shigellosis a therapeutically
effective amount of a compound of claim 41.
58. The compounds:
3-[2-(2-Benzyloxycarbonylamino-3-methyl-butyrylamino)-p-
ropionylamino ]4-oxo-5-(2-phenyl-ethanesulfonylamino)-pentanoic
acid;
3-[2-(2-Benzyloxycarbonylamino-4-carboxy-butyrylamino)-3-methyl-butyrylam-
ino]-4-oxo-5-(2-phenyl-ethanesulfonylamino)-pentanoic acid;
3-{2-[4-Carboxy-2-(3-phenyl-propionylamino)-butyrylamino]-3-methyl-butyry-
lamino}-4-oxo-5-(2-phenyl-ethanesulfonylamino)-pentanoic acid;
3-[2-(2-Benzyloxycarbonylamino-3-methyl-butyrylamino)
propionylamino]-5-(7,7-dimethyl-2-oxo-bicyclo[2.2.1]hept-1-ylmethanesulfo-
nylamino)-4-oxo-pentanoic acid;
3-[2-(2-Benzyloxycarbonylamino-4-carboxy-b-
utyrylamino)-3-methyl-butyrylamino]-5-(7,7dimethyl-2-oxo-bicyclo[2.2.1]hep-
t-1-ylmethanesulfonylamino)-4-oxo-pentanoic acid;
3-{2-[4-Carboxy-2-(3-phe-
nyl-propionylamino)-butyrylamino]-3-methyl-butyrylamino}-5-(7,7-dimethyl-2-
-oxo-bicyclo[2.2.1]hept-1-ylmethanesulfonylamino)-4-oxo-pentanoic
acid;
3-(2-{2-[2-Acetylamino-3-(4-hydroxy-phenyl)-propionylamino]-4-carboxy-but-
yrylamino}-3-methyl-butyrylamino)-5-(7,7-dimethyl-2-oxo-bicyclo[2.2.1]hept-
-1-ylmethanesulfonylamino)-4-oxo-pentanoic acid; and
3-(2-{2-[2-Acetylamino-3-(4-hydroxy-phenyl)-propionylamino]-4-carboxy-but-
yrylamino}-3-methyl-butyrylamino)-4-oxo-5-(2-phenyl-ethanesulfonylamino)-p-
entanoic acid.
Description
FIELD OF THE INVENTION
[0001] This invention relates to compounds that are inhibitors of
interleukin-1.beta. converting enzyme. This invention also relates
to a method of treatment of stroke, reperfusion injury, Alzheimer's
disease, shigellosis, and inflammatory diseases and to a
pharmaceutically acceptable composition that contains a compound
that is an inhibitor of interleukin-1.beta. converting enzyme
(Caspase-1).
BACKGROUND OF THE INVENTION
[0002] The compounds of the present invention are inhibitors of
interleukin-1.beta. converting enzyme (ICE) and are useful in
treating diseases in which interleukin-1 plays a role.
[0003] ICE acts on pro-interleukin-1.beta. (pro-IL-1.beta.) to
produce interleukin-1.beta. (IL-1.beta.), which is an inflammatory
cytokine. In addition, ICE (Caspase-1) regulates at least four
cytokines. ICE activates IL-.beta. and IL-18, and indirectly
regulates the production of IL-1.varies. and IFN.gamma.. Several
diseases are associated with excessive interleukin-1 activity.
Examples of diseases in which interleukin-1 is involved include,
but are not limited to, inflammatory diseases such as rheumatoid
arthritis and inflammatory bowel disease, and neuroinflammatory
disorders such as stroke. Other diseases include septic shock,
reperfusion injury, Alzheimer's disease, and shigellosis.
[0004] Agents that modulate IL-1.beta. activity have been shown to
have beneficial in vivo effects. For example, compounds that are
interleukin-1 receptor antagonists have been shown to inhibit
ischemic and excitotoxic damage in rat brains. See, for example,
Relton J. K., et al., Brain Research Bulletin, 1992;29:243-246.
Additionally, ICE inhibitors were shown to reduce inflammation and
pyrexia in rats. See Elford P. R., et al., British Journal of
Pharmacology, 1995;115:601-606.
[0005] The compounds of the present invention are also inhibitors
of other cysteine proteases in the ICE family. Many of these
proteases have only recently been described in the literature.
While the nomenclature is still unresolved, the following proteases
are representative members of this class of enzymes; Ich-2 (also
called Tx or ICErel-II), ICErel-III, Ich-I (also called Nedd-2),
CPP-32 (also called apopain and yama), Mch-2, Mch-3 (also called
ICE-lap3, CMH-1), and Ced-3. See Henkart P. A., Immunity,
1996;4:195-201. It is recognized that members of this enzyme family
play key biological roles in both inflammation and apoptosis
(programmed cell death). In particular, Caspase-4 can activate
IL-1.beta. and IL-18. It has been shown that a murine homolog of
Caspase-4 can activate ICE. Thus, inhibition of Caspase-4 will act
to inhibit ICE. See Thornberry N. A., et al., Perspectives in Drug
Discovery and Design, 1994;2:389-399.
[0006] In addition to its effects on IL-1.beta. production, ICE has
been shown to play a role in the production of the inflammatory
mediator interferon-.gamma. (Ghayur, et al., Nature,
1997;386(6625):619-623). ICE processes the inactive proform of
interferon-.gamma. inducing factor (IGIF; Interleukin-18) to active
IGIF, a protein which induces production of interferon-.gamma. by
T-cells and natural killer cells. Interferon-.gamma. has been
implicated in the pathogenesis of diseases such as inflammatory
disorders and septic shock. Therefore, ICE inhibitors would be
expected to have beneficial effects in such disease states by
effects on interferon-.gamma..
[0007] Recently, the nomenclature of these cysteine proteases in
the ICE family (also known as Caspases with ICE being known as
Caspase-1) has been further defined. The following proteases are
representative members of this class of enzymes using the
nomenclature described in Alnemri, et al., Cell, 1996;87:171:
Caspase-2 (also known as Ich-1); Caspase-3 (also known as CPP32,
Yama, and apopain); Caspase-4 (also known as TX, Ich2, and ICE
rel-II); Caspase-5 (also known as ICE rel-III); Caspase-6 (also
known as Mch2); Caspase-7 (also known as Mch3); Caspase-8 (also
known as FLICE and Mch5); Caspase-9 (also known as ICE-LAP6 and
Mch6); Caspase-10 (also known as Mch4).
SUMMARY OF THE INVENTION
[0008] The present invention provides compounds of Formula I 1
[0009] R.sup.3 is hydrogen,
[0010] C.sub.1-C.sub.6 alkyl,
[0011] --(CH.sub.2).sub.n aryl, or
[0012] --(CH.sub.2).sub.n heteroaryl;
[0013] R.sup.4 is C.sub.1-C.sub.6 alkyl,
[0014] --(CH.sub.2).sub.n aryl, or
[0015] --(CH.sub.2).sub.n heteroaryl;
[0016] R.sup.5 and R.sup.6 are each independently hydrogen,
[0017] C.sub.1-C.sub.6 alkyl,
[0018] --(CH.sub.2).sub.n aryl, or
[0019] --(CH.sub.2).sub.n heteroaryl;
[0020] R.sup.7 is C.sub.1-C.sub.6 alkyl,
[0021] --(CH.sub.2).sub.n aryl, or
[0022] --(CH.sub.2).sub.n heteroaryl;
[0023] each n is independently 0 to 6;
[0024] each m is independently 0, 1, 2, or 3;
[0025] A is alanine, leucine, isoleucine, proline, phenylalanine,
glycine, tyrosine, serine, threonine, tryptophan, cysteine,
methionine, valine, asparagine, glutamine, aspartic acid, lysine,
glutamic acid, arginine, or histidine;
[0026] each R.sup.Q is independently hydrogen or C.sub.1-C.sub.6
alkyl;
[0027] R.sup.2 is --(CH.sub.2).sub.n--Z; and
[0028] Z is aryl, heteroaryl, cycloalkyl, C.sub.1-C.sub.6alkyl,
2
[0029] fluorenyl, substituted fluorenyl, substituted aryl,
substituted heteroaryl, or substituted cycloalkyl, and the
pharmaceutically acceptable salts, esters, amides, and prodrugs
thereof.
[0030] In a preferred embodiment of the compounds of Formula I,
R.sup.1 is 3
[0031] In another preferred embodiment of the compounds of Formula
I, R.sup.1 is 4
[0032] m is 0, and R.sup.7 is --(CH.sub.2).sub.n aryl.
[0033] In another preferred embodiment of the compounds of Formula
I, R.sup.1 is 5
[0034] m is 0, and R.sup.7is --CH.sub.2aryl.
[0035] In another preferred embodiment of the compounds of Formula
I, R.sup.2 is --(CH2)n aryl.
[0036] In another preferred embodiment of the compounds of Formula
I, aryl is phenyl or naphthyl.
[0037] In another preferred embodiment of the compounds of Formula
I, R.sup.2 is --(CH.sub.2).sub.n-cycloalkyl.
[0038] In another preferred embodiment of the compounds of Formula
I, R.sup.1 is 6
[0039] or --SO.sub.2-phenyl.
[0040] In another preferred embodiment of the compounds of Formula
I, R.sup.2 is 7
[0041] In another preferred embodiment of Formula I, R.sup.2 is
8
[0042] The present invention also provides compounds of the Formula
I 9
[0043] Wherein R.sup.2 is --CH.sub.2CH.sub.2-- aryl, --CH.sub.2--
cycloalkyl, --CH.sub.2CH.sub.2-- cycloalkyl, or
--CH.sub.2CH.sub.2-- heteroaryl;
[0044] R.sup.1 is 10
[0045] R.sup.a is --(CH.sub.2).sub.n-- aryl or --(CH.sub.2).sub.n
heteroaryl;
[0046] R.sup.b is aryl or heteroaryl;
[0047] R.sup.c is --CH.sub.2 aryl or aryl;
[0048] R.sup.d is hydrogen or C.sub.1-C.sub.6 alkyl;
[0049] R.sup.e is --CH.sub.2 aryl or --CH.sub.2 heteroaryl; and the
pharmaceutically acceptable salts, esters, amides, and prodrugs
thereof.
[0050] In a preferred embodiment of the compounds of Formula I,
R.sup.1 is 11
[0051] In another preferred embodiment of the compounds of Formula
I, R.sup.1 is 12
[0052] In another preferred embodiment of the compounds of Formula
I, R.sup.e is --(CH2).sub.n aryl.
[0053] In another preferred embodiment of the compounds of Formula
I, aryl is phenyl or naphthyl.
[0054] In another preferred embodiment of the compounds of Formula
I, R.sup.b is aryl.
[0055] In a preferred embodiment, the present invention provides
the compounds:
[0056]
3-Benzyloxycarbonylamino-4-oxo-5-(2-phenylethanesulfonylamino)-pent-
anoic acid;
[0057]
3-Benzyloxycarbonylamino-4-oxo-5-(3-phenyl-propane-1-sulfonylamino)-
-pentanoic acid;
[0058]
3-Benzyloxycarbonylamino-4-oxo-5-phenylmethanesulfonyl-amino-pentan-
oic acid;
[0059]
5-Benzenesulfonylamino-3-benzyloxycarbonylamino-4-oxo-pentanoic
acid;
[0060]
3-Benzyloxycarbonylamino-5-methanesulfonylamino-4-oxo-pentanoic
acid;
[0061]
3-Benzyloxycarbonylamino-5-(naphthalene-1-sulfonylamino)-4-oxo-pent-
anoic acid;
[0062]
3-Benzyloxycarbonylarnino-5-(2-cyclohexyl-ethanesulfonylamino)-4-ox-
o-pentanoic acid;
[0063]
3-Benzyloxycarbonylamino-5-(2-naphthalen-1-yl-ethanesulfonylamino)--
4-oxo-pentanoic acid;
[0064]
3-Benzyloxycarbonylamino-5-(7,7-dimethyl-2-oxo-bicyclo[2.2.1]hept-1-
-(R)-ylmethanesulfonylamino)-4-oxo-pentanoic acid;
[0065]
3-Benzyloxycarbonylamino-5-(indan-1-ylmethanesulfonylamino)-4-oxo-p-
entanoic acid;
[0066]
3-Benzyloxycarbonylamino-5-(9-fluoro-9H-fluoren-9-ylmethanesulfonyl-
amino)-4-oxo-pentanoic acid;
[0067]
3-Benzyloxycarbonylanino-5-(7,7-dimethyl-2-oxo-bicyclo[2.2.1]hept-1-
-(S)-ylmethanesulfonylamino)-4-oxo-pentanoic acid;
[0068]
3-[2-(2-Benzyloxycarbonylamino-3-methyl-butyrylamino)-propionylamin-
o ]-4-oxo-5-(2-phenyl-ethanesulfonylamino)-pentanoic acid;
[0069]
3-[2-(2-Benzyloxycarbonylamino-4-carboxy-butyrylamino)-3-methyl-but-
yrylamino ]-4-oxo-5-(2-phenyl-ethanesulfonylamino)-pentanoic
acid;
[0070]
3-{2-[4-Carboxy-2-(3-phenyl-propionylamino)-butyrylamino]-3-methyl--
butyrylamino }-4-oxo-5-(2-phenyl-ethanesulfonylamino)-pentanoic
acid;
[0071]
3-(2-{2-[2-Acetylamino-3-(4-hydroxy-phenyl)-propionylamino]-4-carbo-
xy-butyrylamino}-3-methyl-butyrylamino)-4-oxo-5-(2-phenyl-ethanesulfonylam-
ino)-pentanoic acid;
[0072]
3-(2-Acetylamino-3-methyl-butyrylamino)-5-(7,7-dimethyl-2-oxo-bicyc-
lo [2.2.1]hept-1-(S)-ylmethanesulfonylamino)-4-oxo-pentanoic
acid;
[0073]
3-(2-Acetylamino-propylamino)-5-(7,7-dimethyl-2-oxo-bicyclo[2.2.1]h-
ept-1-(S)-ylmethanesulfonylamino)-4-oxo-pentanoic acid;
[0074]
3-[2-(2-Benzyloxycarbonylamino-3-methyl-butyrylamino)-propionylamin-
o
]-5-(7,7-dimethyl-2-oxo-bicyclo[2.2.1]hept-1-ylmethanesulfonylamino)-4-o-
xo-pentanoic acid;
[0075]
3-{2-[4-Carboxy-2-(3-phenyl-propionylamino)-butyrylamino]-3-methyl--
butyrylamino}-5-(7,7-dimethyl-2-oxo-bicyclo[2.2.1]hept-1-ylmethanesulfonyl-
amino)-4-oxo-pentanoic acid;
[0076]
3-(2-{2-[2-Acetylamino-3-(4-hydroxy-phenyl)-propionylamino]-4-carbo-
xy-butyrylamino}-3-methyl-butyrylamino)-5-(7,7-dimethyl-2-oxo-bicyclo
[2.2.1]hept-1-ylmethanesulfonylamino)-4-oxo-pentanoic acid;
[0077]
3-[2-(2-Benzyloxycarbonylamino-4-carboxy-butyrylamino)-3-methyl-but-
yrylamino]-5-(7,7-dimethyl-2-oxo-bicyclo[2.2.1]hept-1-ylmethanesulfonylami-
no)-4-oxo-pentanoic acid;
[0078]
3-(1,2,3,4-tetrahydro-1-oxo-isoquinoline-2-yl)-acetanino-5-benzenes-
ulfonylamino-4-oxo-pentanoic acid;
[0079]
(S)-5-(Bicyclo[2.2.1]hept-1-ylmethanesulfonylamino)-4-oxo-3-[2-(1-o-
xo-3,4-dihydro-1H-isoquinolin-2-yl)-acetylamino]-pentanoic
acid;
[0080]
(S)-4-Oxo-3-[2-(1-oxo-3,4-dihydro-1H-isoquinolin-2-yl)-acetylamino]-
-5-(2-phenyl-ethanesulfonylamino)-pentanoic acid; and
[0081]
4-Oxo-3-[2-(1-oxo-3,4-dihydro-1H-isoquinolin-2-yl)-acetylamino]-5-p-
henylmethanesulfonylamino-pentanoic acid.
[0082] Also provided is a method of inhibiting interleukin-1.beta.
converting enzyme, the method comprising administering to a patient
in need of inhibition of interleukin-1.beta. converting enzyme a
therapeutically effective amount of a compound of Formula I or
II.
[0083] Also provided is a method of inhibiting Caspase4, the method
comprising administering to a patient in need of Caspase-4
inhibition a Caspase-4 inhibiting amount of a compound of Formula I
or II.
[0084] Also provided is a method of treating stroke, the method
comprising administering to a patient having a stroke or having had
a stroke a therapeutically effective amount of a compound of
Formula I or II.
[0085] Also provided is a method of treating inflammatory diseases,
the method comprising administering to a patient having an
inflammatory disease a therapeutically effective amount of a
compound of Formula I or II.
[0086] In a preferred embodiment, the inflammatory disease is
arthritis.
[0087] In another preferred embodiment, the inflammatory disease is
inflammatory bowel disease.
[0088] Also provided is a pharmaceutically acceptable composition
that contains a compound of Formula I or II.
[0089] Also provided is a method of treating septic shock, the
method comprising administering to a patient having septic shock a
therapeutically effective amount of a compound of Formula I or
II.
[0090] Also provided is a method of treating reperfusion injury,
the method comprising administering to a patient having reperfusion
injury a therapeutically effective amount of a compound of Formula
I or II.
[0091] Also provided is a method of treating Alzheimer's disease,
the method comprising administering to a patient having Alzheimer's
disease a therapeutically effective amount of a compound of Formula
I or II.
[0092] Also provided is a method of treating shigellosis, the
method comprising administering to a patient having shigellosis a
therapeutically effective amount of a compound of Formula I or
II.
[0093] The present invention provides compounds of the Formula II
13
[0094] R.sup.a is --(CH.sub.2).sub.n-- aryl or --(CH.sub.2).sub.n
heteroaryl;
[0095] R.sup.b is aryl or heteroaryl;
[0096] R.sup.c is --CH.sub.2 aryl or aryl;
[0097] R.sup.d is hydrogen or C.sub.1-C.sub.6 alkyl;
[0098] R.sup.e is --CH.sub.2 aryl or --CH.sub.2 heteroaryl; and the
pharmaceutically acceptable salts, esters, amides, and prodrugs
thereof.
[0099] In a preferred embodiment of the compounds of Formula II,
R.sup.1 is 14
[0100] In a preferred embodiment of the compounds of Formula II,
R.sup.1 is 15
[0101] In another preferred embodiment of the compounds of Formula
II, R.sup.e is --(CH.sub.2).sub.n aryl.
[0102] In another preferred embodiment of the compounds of Formula
II, aryl is phenyl or naphthyl.
[0103] In another preferred embodiment of the compounds of Formula
II, R.sup.b is aryl.
DETAILED DESCRIPTION OF THE INVENTION
[0104] The term "alkyl" means a straight or branched chain
hydrocarbon. Representative examples of alkyl groups are methyl,
ethyl, propyl, isopropyl, isobutyl, butyl, tert-butyl, sec-butyl,
pentyl, and hexyl.
[0105] The term "cycloalkyl" means a cyclic alkyl group having 3 to
8 carbons. The cycloalkyl group can be fused to one or more aryl or
heteroaryl groups. Representative examples are cyclopentyl,
cyclohexyl, 1- or 2-indanyl, 1- or 2-tetralinyl, and 9-fluorenyl.
The term "cycloalkyl" includes bicycloalkyl and substituted
bicycloalkyl. Suitable substituents are defined with respect to
aryl below.
[0106] The term "alkoxy" means an alkyl group attached to an oxygen
atom. Representative examples of alkoxy groups include methoxy,
ethoxy, tert-butoxy, propoxy, and isobutoxy.
[0107] The term "halogen" includes chlorine, fluorine, bromine, and
iodine.
[0108] The term "aryl" means an aromatic hydrocarbon.
Representative examples of aryl groups include phenyl, naphthyl,
and biphenyl.
[0109] The term "heteroatom" includes oxygen, nitrogen, sulfur, and
phosphorus.
[0110] The term "heteroaryl" means an aryl group wherein one or
more carbon atom of the aromatic hydrocarbon has been replaced with
a heteroatom. Examples of heteroaryl groups include furan,
thiophene, pyrrole, thiazole, pyridine, pyrimidine, pyrazine,
benzofuran, indole, coumarin, quinoline, isoquinoline, carbazole,
and naphthyridine.
[0111] The aryl or heteroaryl groups may be substituted with one or
more substituents, which can be the same or different. Examples of
suitable substituents include alkyl, alkoxy, thioalkoxy, hydroxy,
halogen, trifluoromethyl, amino, alkylamino, dialkylamino,
--(CH.sub.2).sub.nOH, --NO.sub.2, --CN, --CO.sub.2H,
--CO.sub.2alkyl, --SO.sub.3H, --CHO, --COalkyl, --CONH.sub.2,
--CONH-alkyl, --CONHR.sup.q, --CON(alkyl).sub.2,
--(CH.sub.2).sub.n--NH.sub.2, --(CH.sub.2).sub.n--NH-alkyl,
--NHR.sup.q, or --NHCOR.sup.q, where n is 1 to 5 and R.sup.q is
hydrogen or alkyl. It is intended that the terms "aryl" and
"heteroaryl" include unsubstituted as well as substituted aryl and
heteroaryl groups. It is also intended that the substituents on the
aryl or heteroaryl groups include other cyclic compounds that are
fused to the aryl or heteroaryl groups, typically by adjacent
carbon atoms. For example, a phenyl group may be fused with a
cyclohexane group.
[0112] The symbol "--" means a bond.
[0113] The compounds of Formula I or II can be administered to a
patient either alone or as part of a pharmaceutically acceptable
composition. The compositions can be administered to patients such
as humans and animals either orally, rectally, parenterally
(intravenously, intramuscularly, or subcutaneously),
intracisternally, intravaginally, intraperitoneally,
intravesically, locally (powders, ointments or drops), or as a
buccal or nasal spray.
[0114] Compositions suitable for parenteral injection may comprise
physiologically acceptable sterile aqueous or nonaqueous solutions,
dispersions, suspensions or emulsions, and sterile powders for
reconstitution into sterile injectable solutions or dispersions.
Examples of suitable aqueous and nonaqueous carriers, diluents,
solvents or vehicles include water, ethanol, polyols
(propyleneglycol, polyethyleneglycol, glycerol, and the like),
suitable mixtures thereof, vegetable oils (such as olive oil) and
injectable organic esters such as ethyl oleate. Proper fluidity can
be maintained, for example, by the use of a coating such as
lecithin, by the maintenance of the required particle size in the
case of dispersions and by the use of surfactants.
[0115] These compositions may also contain adjuvants such as
preserving, wetting, emulsifying, and dispensing agents. Prevention
of the action of microorganisms can be ensured by various
antibacterial and antifungal agents, for example, parabens,
chlorobutanol, phenol, sorbic acid, and the like. It may also be
desirable to include isotonic agents, for example sugars, sodium
chloride, and the like. Prolonged absorption of the injectable
pharmaceutical form can be brought about by the use of agents
delaying absorption, for example, aluminum monostearate and
gelatin.
[0116] Solid dosage forms for oral administration include capsules,
tablets, pills, powders, and granules. In such solid dosage forms,
the active compound is admixed with at least one inert customary
excipient (or carrier) such as sodium citrate or dicalcium
phosphate or (a) fillers or extenders, as for example, starches,
lactose, sucrose, glucose, mannitol, and silicic acid, (b) binders,
as for example, carboxymethylcellulose, alignates, gelatin,
polyvinylpyrrolidone, sucrose, and acacia, (c) humectants, as for
example, glycerol, (d) disintegrating agents, as for example,
agar-agar, calcium carbonate, potato or tapioca starch, alginic
acid, certain complex silicates, and sodium carbonate, (e) solution
retarders, as for example paraffin, (f) absorption accelerators, as
for example, quaternary ammonium compounds, (g) wetting agents, as
for example, cetyl alcohol and glycerol monostearate, (h)
adsorbents, as for example, kaolin and bentonite, and (i)
lubricants, as for example, talc, calcium stearate, magnesium
stearate, solid polyethylene glycols, sodium lauryl sulfate, or
mixtures thereof. In the case of capsules, tablets, and pills, the
dosage forms may also comprise buffering agents.
[0117] Solid compositions of a similar type may also be employed as
fillers in soft and hard-filled gelatin capsules using such
excipients as lactose or milk sugar as well as high molecular
weight polyethyleneglycols, and the like.
[0118] Solid dosage forms such as tablets, dragees, capsules,
pills, and granules can be prepared with coatings and shells, such
as enteric coatings and others well known in the art. They may
contain opacifying agents, and can also be of such composition that
they release the active compound or compounds in a certain part of
the intestinal tract in a delayed manner. Examples of embedding
compositions which can be used are polymeric substances and waxes.
The active compounds can also be in micro-encapsulated form, if
appropriate, with one or more of the above-mentioned
excipients.
[0119] Liquid dosage forms for oral administration include
pharmaceutically acceptable emulsions, solutions, suspensions,
syrups, and elixirs. In addition to the active compounds, the
liquid dosage forms may contain inert diluents commonly used in the
art, such as water or other solvents, solubilizing agents and
emulsifiers, as for example, ethyl alcohol, isopropyl alcohol,
ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate,
propyleneglycol, 1,3-butyleneglycol, dimethylformamide, oils, in
particular, cottonseed oil, groundnut oil, corn germ oil, olive
oil, castor oil, and sesame oil, glycerol, tetrahydrofurfuryl
alcohol, polyethyleneglycols, and fatty acid esters of sorbitan or
mixtures of these substances, and the like.
[0120] Besides such inert diluents, the composition can also
include adjuvants, such as wetting agents, emulsifying, and
suspending agents, sweetening, flavoring, and perfuming agents.
[0121] Suspensions, in addition to the active compounds, may
contain suspending agents, as for example, ethoxylated isostearyl
alcohols, polyoxyethylene sorbitol and sorbitan esters,
microcrystalline cellulose, aluminum metahydroxide, bentonite,
agar-agar and tragacanth, or mixtures of these substances, and the
like.
[0122] Compositions for rectal administrations are preferably
suppositories which can be prepared by mixing the compounds of the
present invention with suitable non-irritating excipients or
carriers such as cocoa butter, polyethyleneglycol, or a suppository
wax, which are solid at ordinary temperatures but liquid at body
temperature and therefore, melt in the rectum or vaginal cavity and
release the active component.
[0123] Dosage forms for topical administration of a compound of
this invention include ointments, powders, sprays, and inhalants.
The active component is admixed under sterile conditions with a
physiologically acceptable carrier and any preservatives, buffers,
or propellants as may be required. Ophthalmic formulations, eye
ointments, powders and solutions are also contemplated as being
within the scope of this invention.
[0124] The compounds of the present invention can be administered
to a patient at dosage levels in the range of about 0.1 to about
1,000 mg per day. For a normal human adult having a body weight of
about 70 kg, a dosage in the range of about 0.01 to about 100 mg/kg
of body weight per day is preferable. The specific dosage used,
however, can vary. For example, the dosage can depended on a
numbers of factors including the requirements of the patient, the
severity of the condition being treated, and the pharmacological
activity of the compound being used. The determination of optimum
dosages for a particular patient is well known to those skilled in
the art.
[0125] The term "pharmaceutically acceptable salts, esters, amides,
and prodrugs" as used herein refers to those carboxylate salts,
amino acid addition salts, esters, amides, and prodrugs of the
compounds of the present invention which are, within the scope of
sound medical judgment, suitable for use in contact with the
tissues of patients without undue toxicity, irritation, allergic
response, and the like, commensurate with a reasonable benefit/risk
ratio, and effective for their intended use, as well as the
zwitterionic forms, where possible, of the compounds of the
invention. The term "salts" refers to the relatively non-toxic,
inorganic and organic acid addition salts of compounds of the
present invention. These salts can be prepared in situ during the
final isolation and purification of the compounds or by separately
reacting the purified compound in its free-base form with a
suitable organic or inorganic acid and isolating the salt thus
formed. Representative salts include the hydrobromide,
hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate,
valerate, oleate, palmitate, stearate, laurate, borate, benzoate,
lactate, phosphate, tosylate, citrate, maleate, fumarate,
succinate, tartrate, naphthylate mesylate, glucoheptonate,
lactobionate, and laurylsulphonate salts and the like. These may
include cations based on the alkali and alkaline earth metals, such
as sodium, lithium, potassium, calcium, magnesium, and the like, as
well as nontoxic ammonium, quaternary ammonium, and amine cations
including, but not limited to ammonium, tetramethylammonium,
tetraethylammonium, methylamine, dimethylamine, trimethylamnine,
triethylamine, ethylamine, and the like. (See, for example, Berge
S. M., et al., "Pharmaceutical Salts," J. Pharm. Sci., 1977;66:1-19
which is incorporated herein by reference.)
[0126] Examples of pharmaceutically acceptable, non-toxic esters of
the compounds of this invention include C.sub.1-C.sub.6 alkyl
esters wherein the alkyl group is a straight or branched chain.
Acceptable esters also include C.sub.5-C.sub.7 cycloalkyl esters as
well as arylalkyl esters such as, but not limited to benzyl.
C.sub.1-C.sub.4 alkyl esters are preferred. Esters of the compounds
of the present invention may be prepared according to conventional
methods.
[0127] Examples of pharmaceutically acceptable, non-toxic amides of
the compounds of this invention include amides derived from
ammonia, primary C.sub.1-C.sub.6 alkyl amines, and secondary
C.sub.1-C.sub.6 dialkyl amines wherein the alkyl groups are
straight or branched chain. In the case of secondary amines the
amine may also be in the form of a 5- or 6-membered heterocycle
containing one nitrogen atom. Amides derived from ammonia,
C.sub.1-C.sub.3 alkyl primary amines, and C.sub.1-C.sub.2 dialkyl
secondary amines are preferred. Amides of the compounds of the
invention may be prepared according to conventional methods.
[0128] The term "prodrug" refers to compounds that are rapidly
transformed in vivo to yield the parent compound of the above
formulae, for example, by hydrolysis in blood. A thorough
discussion is provided in T. Higuchi and V. Stella, "Pro-drugs as
Novel Delivery Systems," Vol. 14 of the A.C.S. Symposium Series,
and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche,
American Pharmaceutical Association and Pergamon Press, 1987, both
of which are incorporated herein by reference.
[0129] In addition, the compounds of the present invention can
exist in unsolvated as well as solvated forms with pharmaceutically
acceptable solvents such as water, ethanol, and the like. In
general, the solvated forms are considered equivalent to the
unsolvated forms for the purposes of the present invention.
[0130] The compounds of the present invention can exist in
different stereoisomeric forms by virtue of the presence of
asymmetric centers in the compounds; ie, each asymmetric carbon can
have either the R or S configuration. It is contemplated that all
stereoisomeric forms of the compounds as well as mixtures thereof,
including racemic mixtures, form part of this invention.
[0131] The compounds of the present invention are administered to a
patient in need of ICE inhibition. In general, patients in need of
ICE inhibition are those patients having a disease or condition in
which interleukin-1 plays a role. Examples of such diseases
include, but are not limited to, inflammatory diseases such as
rheumatoid arthritis, inflammatory bowel disease, and
neuroinflammatory disorders such as stroke and septic shock. Other
diseases include reperfusion injury, Alzheimer's disease, and
shigellosis.
[0132] A "therapeutically effective amount" is an amount of a
compound of Formula I or II that when administered to a patient
having a disease that can be treated with a compound of Formula I
or II ameliorates a symptom of the disease. A therapeutically
effective amount of a compound of Formula I or II is readily
determined by one skilled in the art by administering a compound of
Formula I or II to a patient and observing the results.
[0133] The following examples illustrate particular embodiments of
the invention and are not intended to limit the scope of the
specification and claims in any manner.
[0134] Compounds of the current invention can be prepared generally
by converting the appropriate starting sulfonamide 1 to Boc
sulfonamide 2 using a reagent such as di-tert-butyl dicarbonate.
Boc sulfonamide 2 may then be reacted with the appropriately
substituted aspartic acid bromomethylketone .beta. tert-butyl ester
3 in the presence of a base, followed by treatment with acid to
give the desired product 4. 16
[0135] Alternatively, compounds of the current invention can be
prepared generally by reaction of the appropriately substituted
aspartic acid aldehyde 1 with nitromethane in the presence of a
base such as potassium tert-butoxide to give nitro alcohol 2.
Reduction of 2 to the amine 3, followed by reaction with the
appropriate sulfonyl chloride gives 4 which may be oxidized to the
ketone 5 with a reagent such as Dess Martin periodinane or by a
Swern oxidation. Acidic deprotection of the t-butyl ester with HCI
or trifluoroacetic acid gives the desired product 6. 17
EXAMPLE 1a
1,1-Dimethylethyl [(2-phenylethyl)sulfonyl]carbamate
[0136] A solution of di-tert-butyldicarbonate (1.07 g) in methylene
chloride (3 mL) was added dropwise to a solution of
2-phenylethanesulfonamide (0.78 g), triethylamine (0.48 g), and
DMAP (dimethylamino-pyridine) (0.012 g) in methylene chloride (10
mL). The reaction mixture was stirred for 3 hours at ambient
temperature and the solvent was evaporated. The resulting oil was
taken up in ethyl acetate, washed with 5% HCl, water, and then
brine. The organic layer was dried over sodium sulfate, filtered,
and the solvent evaporated. The resulting oil was purified by flash
column chromatography on silica (1% ether/10% hexane/methylene
chloride gradient to 20% ether/10% hexane/methylene chloride) to
yield 1,1-dimethylethyl [(2-phenylethyl)sulfonyl]carbamate (0.87
g), melting point (mp) 99-102.degree. C., which was used in the
next step without further purification.
[0137] The following compounds were prepared according to the
procedure of Example 1a from the corresponding sulfonamides:
EXAMPLE 1b
1,1-Dimethylethyl [(3-phenylpropyl)sulfonyl]carbamate, mp
59-62.degree. C.
EXAMPLE 1c
1,1-Dimethylethyl [(phenylmethyl)sulfonyl]carbamate, mp
90-94.degree. C. (dec).
EXAMPLE 1d
Tert-butyl[benzenesulfonyl]carbamate
[0138] MS (AP-): 256.
[0139] CHN Calculated: C, (51.35%); H, (5.88%), N, (5.44%/0); S,
(12.46%).
[0140] Found: C, (51.41%); H, (5.59%); N, (5.40%); S, (12.44%).
EXAMPLE 1e
Tert-butyl[methanesulfonyl]carbamate
[0141] MS (AP-): 194.
[0142] CHN Calculated: C, (36.91%); H, (6.71%); N, (7.17%); S,
(16.42%). Found: C, (36.96%); H, (6.54%); N, (7.08%); S,
(16.39%).
EXAMPLE 1f
Tert-butyl[naphthalene-1-sulfonyl]carbamate
[0143] MS (AP-): 306. CHN Calculated: C, (58.62%); H, (5.57%); N,
(4.56%); S, (10.43%). Found: C, (58.54%); H, (5.40%); N, (4.44%);
S, (1 0.40%).
EXAMPLE 1g
Tert-butyl[2-cyclohexyl-ethanesulfonyl]carbamate
[0144] MS (AP-): 290.
[0145] CHN Calculated: C, (53.58%); H, (8.65%); N, (4.81%); S
(11.00%).
[0146] Found: C, (53.64%); H, (8.58%); N, (4.89%); S (11.26%).
EXAMPLE 1h
Tert-butyl[2-naphthalen-1-yl -ethanesulfony]carbamate
[0147] NMR (CDCl.sub.3): 7.98 (d, 1H), 7.89 (d, 1H), 7.79 (d, 1H),
7.6-7.3 (m, 4H), 6.85 (br, 1H), 3.83-3.77 (m, 2H), 3.64-3.60 (m,
2H), 1.44 (s, 9H).
Example 1i
Tert-butyl[7,7-dimethyl-2-oxo-bicyclo[2.2.1]hept-1-ylmethanesulfonyl]carba-
mate
[0148] MS (AP+): base peak 276 (parent with loss of isobutylene).
CHN Calculated: C, (54.36%); H, (7.60%); N, (4.23%). Found: C,
(54.64%); H, (7.62%); N, (3.98%).
EXAMPLE 1j
Tert-butyl[indan-1-ylmethanesulfonyl]carbamate
[0149] IR (cm.sup.-1): 3246, 3232, 2980, 2937, 1740, 1436, 1350,
1243, 1137, 9176, 830, 757.
[0150] MS (AP+): 310.
EXAMPLE 1k
Tert-butyl[9-fluoro-9H-fluoren-9-ylmethanesulfonyl]carbamate
[0151] MS (AP-): 376.
[0152] CHN Calculated: C, (60.46%); H, (5.34%); N, (3.71%); S,
(8.50%); F, (5.03%).
[0153] Found: C, (60.19%); H, (5.40%); N, (3.64%); S, (8.33%); F,
(4.89%).
EXAMPLE 2a
3-Benzyloxycarbonylamino-4-oxo-5-(2-phenylethane-sulfonylamino)-pentanoic
acid
[0154] To a solution of 1,1-dimethylethyl
[(2-phenylethyl)sulfonyl]carbama- te (0.28 g) in dry DMF
(dimethylformamide) (2 mL) was added potassium tert-butoxide (0.12
g) and the resulting solution was added dropwise to an ice-cooled
solution of 3-benzyloxycarbonylamino-5-bromo-4-oxo-pentanoi- c acid
1,1-dimethylethyl ester (0.32 g) in DMF (2 mL). The reaction
mixture was stirred at room temperature for 24 hours, poured into
water (100 mL), and the resulting solution was neutralized with
dilute aqueous NH.sub.4Cl. The product was extracted into ether
(3.times.50 mL) and the combined organic layers were washed with
water, dilute Na.sub.2S.sub.2O.sub.3, and then brine. The solution
was dried over sodium sulfate, filtered, and the solvent was
evaporated to give the crude intermediate ester (0.49 g) as a
yellow oil.
[0155] The oil was dissolved in methylene chloride (10 mL) and
trifluoroacetic acid (10 mL) and the resulting solution was stirred
at room temperature for 6 hours. The solvent was evaporated to give
a yellow oil which was purified by column chromatography (silica;
1% acetone/1% formic acid/methylene chloride gradient to 20%
acetone/1% formic acid/methylene chloride) and recrystallized from
ether/hexane to give
3-benzyloxycarbonylamino-4-oxo-5-(2-phenyl-ethanesulfonylamino)-pentanoic
acid (0.04 g), mp 100-101.degree. C. (NMR [CD.sub.3OD], ppm)
7.4-7.1 (m, 10H), 5.1 (s, 2H), 4.5 (t, 1H), 4.35 (d, 1H), 4.15 (d,
1H), 3.3-3.0 (m, 4H), 2.9-2.7 (m, 2H).
[0156] The following compounds were prepared according to the
procedure of Example 2a from the corresponding Boc protected
sulfonamides:
EXAMPLE 2b
3-Benzloxycarbonylamino-4-oxo-5-(3-phenylpropane-1-sulfonylamino)-pentanoi-
c acid, mp 96-104.degree. C.
[0157] (NMR [CD.sub.3OD], ppm) 7.4-7.1 (m, 10H), 5.11 (s, 2H), 4.5
(t, 1H), 4.3 (d, 1H), 4.1 (d, 1H), 3.05-2.6 (m, 6H), 2.2-2.0 (m,
2H).
EXAMPLE 2c
3-Benzyloxycarbonylamino-4-oxo-5-phenylmethanesulfonyl-amino-pentanoic
acid mp 160-164.degree. C.
[0158] (NMR [CD.sub.3OD], ppm) 7.5-7.2 (m, 10H), 5.13 (s, 2H), 4.47
(t, 1H), 4.30 (s, 2H), 4.20 (d, 1H), 4.0 (d, 1H), 2.9-2.7 (m,
2H).
EXAMPLE 2d
5-Benzenesulfonylamino-3-benzyloxycarbonylamino-4-oxo-pentanoic
acid
[0159] MS (AP+): 421.
[0160] CHN Calculated: C, (54.28%); H, (4.79%); N, (6.66%); S,
(7.63%).
[0161] Found: C, (54.19%); H, (4.85%); N, (6.47%); S, (7.36%).
[0162] Water (0.10%).
EXAMPLE 2e
3-Benzyloxycarbonylamino-5-methanesulfonylamino-4-oxo-pentanoic
acid
[0163] NMR (ppm, CD.sub.3OD): 7.4-7.2 (m, 5H), 5.48 (s, 2H), 4.51
(t, 1H), 4.4-4.1 (dd, 2H), 2.9-2.7 (m, 5H).
[0164] CHN Calculated: C, (53.58%); H, (8.65%); N, (4.81%); S,
(11.00%).
[0165] Found: C, (53.64%); H, (8.58%); N, (4.89%); S, (11.26%).
EXAMPLE 2f
3-Benzyloxycarbonylamino-5-(naphthalene-1-sulfonylamino)-4-oxo-pentanoic
acid
[0166] MS (AP+): 471.
[0167] IR (KBr, cm.sup.-1): 3347, 2928, 1717, 1508, 1327, 1162,
1134, 772, 589.
EXAMPLE 2g
3-Benzyloxycarbonylamino-5-(2-cyclohexyl-ethanesulfonylamino)-4-oxo-pentan-
oic acid
[0168] NMR (ppm, CD.sub.3OD): 7.4-7.2 (m, 5H), 5.12 (s, 2H), 4.50
(t, 1H), 4.4-4.2 (d, 1H), 4.2-4.0 (d, 1H), 3.1-2.7 (m, 4H), 1.8-1.6
(m, 8H), 1.4-1.1 (m, 5H), 1.0-0.8 (m, 2H).
EXAMPLE 2h
3-Benzyloxycarbonylamino-5-(2-naphthalen-1-yl-ethanesulfonylamino)-4-oxo-p-
entanoic acid
[0169] NMR (ppm, CD.sub.3OD) 8.11 (d, 1H), 7.88 (d, 1H), 7.77 (d,
1H), 7.6-7.2 (m, 9H), 5.06 (s, 2H), 4.51 (t, 1H), 4.4 (d, 1H), 4.2
(d, 1H), 3.7-3.5 (m, 2H), 3.4-3.3 (m), 2.9-2.7 (m, 2H). IR (KBr,
cm.sup.-1): 3307, 2926, 1735, 1685, 1544, 1398, 1322, 1275, 1136,
778, 698.
EXAMPLE 2i
3-Benzyloxycarbonylamino-5-(7,7-dimethyl-2-oxo-bicyclo[2.2.1]hept-1-(R)-yl-
methanesulfonylamino)-4-oxo-pentanoic acid
[0170] IR (LF+CHCl.sub.3, cm.sup.-1): 3314, 2960, 1730.5, 1525,
1329, 1217, 1146, 1052, 754.
[0171] NMR (CD.sub.3OD, ppm): 7.4-7.2 (m, 5H), 5.12 (s, 2H), 4.5
(t, 1H), 4.4 (d, 1H), 4.15 (d, 1H), 3.45 (d,1H), 3.0 (d, 1H),
2.9-2.6 (m, 2H), 2.5-2.3 (m, 2H), 2.2-1.8 (m, 3H), 1.8-1.6 (m, 1H),
1.5-1.4 (m, 1H), 1.06 (s, 3H), 0.87 (s, 3H).
EXAMPLE 2j
3-Benzyloxycarbonylamino-5-(indan-1-ylmethanesulfonylamino)-4-oxo-pentanoi-
c acid
[0172] IR(KBr, cm.sup.-1): 3314, 2930, 1704, 1530, 1318, 1266,
1145, 1059, 746, 698. MS (AP+): 475.
EXAMPLE 2k
3-Benzyloxycarbonylamino-5-(9-fluoro-9H-fluoren-9-ylmethanesulfonylamino)--
4-oxo-pentanoic acid
[0173] MS (AP+): 521 (parent with loss of F-).
[0174] NMR (F19, CD.sub.3OD, ppm)-77.1.
EXAMPLE 2l
3-Benzyloxycarbonylamino-5-(7.7-dimethyl-2-oxo-bicyclo[2.2.1]hept-1-(S)-yl-
methanesulfonylamino)-4-oxo-pentanoic acid
[0175] MS (AP-): 493.
[0176] IR (KBr, cm.sup.-1): 3374, 2961, 1733, 1522, 1455, 1416,
1330, 1274, 1204, 1179, 1146, 1052, 698.
EXAMPLE 2m
(R)-3-Benzyloxycarbonylamino-5-(7,7-dimethyl-bicyclo[2.2.1]hept-1-ylmethan-
e-sulfonylamino)-4-oxo-pentanoic acid
[0177] MS (APCI-): 479.2
[0178] The following compounds were prepared according to the
procedure of Example 2a from the corresponding di, tri or
tetrapeptide bromomethylketones and
tert-butyl[(2-phenylethyl)sulfonyl]carbamate:
EXAMPLE 3a
3-[2-(2-Benzyloxycarbonylamino-3-methyl-butyrylamino)-propionylamino]-4-ox-
o-5-(2-phenyl-ethanesulfonylamino)-pentanoic acid
[0179] Calculated for
C.sub.29H.sub.38N.sub.4O.sub.9S.sub.1.multidot.0.33 CF.sub.3COOH:
C, 54.26; H, 5.88; N, 8.53. Found: C, 54.26; H, 5.93; N, 8.55.
Example 3b
3-[2-(2-Benzyloxycarbonylamino-4-carboxy-butyrylamino)-3-methyl-butyrylami-
no]-4-oxo-5-(2-phenyl-ethanesulfonylamino)-pentanoic acid
EXAMPLE 3c
3-{2-[4-Carboxy-2-(3-phenyl-propionylamino)-butyrylamino]-3-methyl-butyryl-
amino}-4-oxo-5-(2-phenyl-ethanesulfonylamino)-pentanoic acid
[0180] Calculated for
C.sub.32H.sub.42N.sub.4O.sub.10S.sub.1.multidot.0.24 CF.sub.3COOH:
C, 55.56; H, 6.06; N, 7.98.
[0181] Found: C, 55.56; H, 6.23; N, 8.07.
EXAMPLE 3d
3-2-{2-[2-Acetylamino-3-(4-hydroxy-phenyl)-propionylamino]-4-carboxy-butyr-
ylamino
}-3-methyl-butyrylamino)-4-oxo-5-(2-phenyl-ethanesulfonylamino)-pe-
ntanoic acid
[0182] The following compounds were prepared according to the
procedure of Example 2a from the corresponding protected di, tri or
tetrapeptide bromomethylketones and
tert-butyl[7,7-dimethyl-2-oxo-bicyclo[2.2.1]hept-1-
-ylmethanesulfonyl]carbamate:
EXAMPLE 4a
3-(2-Acetylamino-3-methyl-butyrylamino)-5-(7,7-dimethyl-2-oxo-bicyclo
[2.2.1]hept-1-(S)-ylmethanesulfonylamino)-4-oxo-pentanoic acid
[0183] MS (AP+): 502.
[0184] IR (KBr, cm.sup.-1): 3338, 2965, 1738, 1653, 1540, 1395,
1328, 1148.
EXAMPLE 4b
3-(2-Acetylamino-propylamino)-5-(7,7-dimethyl-2-oxo-bicyclo[2.2.1]hept-1-(-
S)-ylmethanesulfonylamino)-4-oxo-pentanoic acid
[0185] MS (AP-): 472.
Example 4c
3-[2-(2-Benzyloxycarbonylamino-3-methyl-butyrylamino)-propionylamino]-5-(7-
,7-dimethyl-2-oxo-bicyclo[2.2.1]hept-1-ylmethanesulfonylamino)-4-oxo-penta-
noic acid
[0186] Calculated for
C.sub.31H.sub.44N.sub.4O.sub.10S.sub.1.multidot.0.30 CF.sub.3COOH:
C, 54.31; H, 6.39; N, 8.02.
[0187] Found: C, 54.31; H, 6.51; N, 7.80.
EXAMPLE 4d
3-{2-[4-Carboxy-2-(3-phenyl-propionylamino-butyrylamino]-3-methyl-butyrlam-
ino}-5-(7,7-dimethyl-2-oxo-bicyclo[2.2.1]hept-1-ylmethanesulfonylamino)-4--
oxo-pentanoic acid
[0188] Calculated for
C.sub.34H48N.sub.4O.sub.11S.sub.1.multidot.0.43 CF.sub.3COOH: C,
54.36; H, 6.34; N, 7.27.
[0189] Found: C, 54.36; H, 6.57; N, 7.35.
EXAMPLE 4e
3-(2-{2-[2-Acetylamino-3-(4-hydroxy-phenyl)-propionylamino]-4-carboxy-buty-
rylamino}-3-methyl-butyrylamino)-5-(7,7-dimethyl-2-oxo-bicyclo[2.2.1]hept--
1-ylmethanesulfonylamino)-4-oxo-pentanoic acid
EXAMPLE 4f
3-[2-(2-Benzyloxycarbonylamino-4-carboxy-butyrylamino)-3-methyl-butyrylami-
no]-5-(7,7-dimethyl-2-oxo-bicyclo[2.2.1]hept-1-ylmethanesulfonylamino)-4-o-
xo-pentanoic acid
[0190] The compounds of the present invention can also be
synthesized by the following route: 18
EXAMPLE 5a
3-(1,2,3,4,-Tetrahydro-1-oxo-isoquinoline-2-yl)acetamino-5-benzenesulfonyl
amino-4-oxo-pentanoic acid
[0191] Step A
[0192] To a solution of
(1,2,3,4-tetrahydro-1-oxo-isoquinoline-2-yl)acetic acid (2.7 g,
13.0 mMol) prepared according to the procedure of Anderson W. K.,
et al., J. Med. Chem, 1988;31:2097 and H-Asp (OtBu)OMe.times.HCl
(2.9 g, 12.0 mMol) in dimethylformamide (40 mL) was added at
0.degree. C. 1-ethyl-3-(3'-dimethylamino-propyl)
carbodiimide.times.HCl (2.5 g, 13.0 mMol) and triethylamine (4.05
g, 40 mMol). The mixture was stirred at room temperature for 16
hours. Most of the solvent was removed under reduced pressure, the
residue was dissolved in ethyl acetate. The organic phase was
washed successively with aqueous sodium hydrogen-carbonate and
water, dried over sodium sulfate, and concentrated to give 4.5 g of
an amorphous residue.
[0193] The residue was dissolved in 40 mL of dioxane/water (1: 1)
and hydrolyzed in the presence of thymolphthalein by dropwise
addition of 1N NaOH (12.0 mL). After evaporation of most of the
dioxane and dilution with water the aqueous solution was extracted
with ether, acidified with dilute HCl to pH 2-3, and the product
extracted into ethyl acetate. The organic phase was washed with
water, dried over sodium sulfate, and concentrated under reduced
pressure to give 3.4 g of crystalline
N-(1,2,3,4-tetrahydro-1-oxo-isoquinoline-2-yl)-acetyl aspartic
acid, 4tert-butyl ester.
[0194] Step B
[0195] To a solution of
N-(1,2,3,4-tetrahydro-1-oxo-isoquinoline-2-yl)-ace- tyl aspartic
acid, 4-tert-butyl ester (3.1 g, 8.25 mMol) in 50 mL of
tetrahydrofuran was added at -5.degree. C. N-methylmorpholine (1.25
mL, 11.0 mMol) and isobutylchloroformate (1 eq). After 15 minutes
between -5.degree. C. and 0.degree. C. the formed mixed anhydride
was added at -78.degree. C. to a suspension of sodium borohydride
(0.75 g, 20 mMol) in 45 mL of tetrahydrofuran and 15 mL of
methanol. After 2 hours at -40.degree. C. the reaction was quenched
by addition of 5.0 mL of acetic acid. Ethyl acetate/hexane (250 mL,
1:1) and water 30 mL was added. The organic phase was washed
successively with saturated aqueous sodium hydrogen carbonate and
water, dried over sodium sulfate, and concentrated under reduced
pressure. Purification of the residue over silica (elution with
dichloromethane/methanol 20:1) gave
3-(1,2,3,4-tetrahydro-1-oxo-isoq-
uinoline-2-yl)-acetamino-4-hydroxy-pentanoic acid, tert-butyl ester
(2.3 g).
[0196] Step C
[0197] To a solution at -45.degree. C. under nitrogen of
dimethylsulfoxide (3.72 mL, 52.4 mMol) in dichloromethane was added
dropwise via syringe oxalyl chloride (2.5 g, 28.8 mMol) followed by
N-(1,2,3,4-tetrahydro-1-ox-
o-isoquinoline-2-yl)-acetamino-4-hydroxy-pentanoic acid, tert-butyl
ester (6.88 g, 24.0 mMol). After 30 minutes the reaction was
quenched by addition of diisopropylethyl amine (12.4 mL, 72.0 mMol)
and partitioned between ethyl acetate (800 mL) and water (80 mL).
The organic phase was washed successively with 1N sodium hydrogen
sulfate and water, dried over sodium sulfate, and concentrated
under reduced pressure to give 4.8 g of
3-(1,2,3,4-tetrahydro-1-oxo-isoquinoline-2-yl)-acetamino-4-oxo-pentanoic
acid, tert-butyl ester.
[0198] Step D
[0199] To 1.15 g (10.0 mMol) of KOtBu (potassium tertbutoxide) in
dimethylformamide (30 mL) at 0.degree. C. under nitrogen and
nitromethane (1.75 mL, 32.5 mMol) was added
3-(1,2,3,4-tetrahydro-1-oxo-isoquinoline-2-
-yl)-acetamino-4-oxo-pentanoic acid, tert-butyl ester (3.6 g, 10.0
mMol). After 3 hours at 0.degree. C. the reaction was quenched by
addition of 1.5 mL of acetic acid and partitioned between ethyl
acetate (200 mL) and water (20 mL). The organic phase was washed
with saturated aqueous sodium hydrogen carbonate and water, dried
over sodium sulfate, and concentrated under reduced pressure.
Purification of the residue over silica (elution with
dichloromethane/methanol 20:1) gave 2.7 g of
3-(1,2,3,4-tetrahydro-1-
-oxo-isoquinoline-2-yl)-acetamino-4-hydroxy-5-nitro-pentanoic acid,
tert-butyl ester as 1:1 mixture of diastereomers.
[0200] Step E
[0201] A mixture of
3-(1,2,3,4-tetrahydro-1-oxo-isoquinoline-2-yl)-acetami-
no-4-hydroxy-5-nitro-pentanoic acid, tert-butyl ester (2.55 g, 6.05
mMol) and 10% Pd on charcoal (1.5 g) in 100 mL of methanol
containing 5 mL of 10% aqueous acetic acid was hydrogenated at room
temperature for 4 hours. Filtration and evaporation of the solvent
under reduced pressure gave of
3-(1,2,3,4-tetrahydro-1-oxo-isoquinoline-2-yl)-acetamino-5-amino-4-hydrox-
y-pentanoic acid, tert-butyl ester hydroacetate (2.7 g).
[0202] Step F
[0203] To a solution of
3-(1,2,3,4-tetrahydro-1-oxo-isoquinoline-2-yl)-ace-
tamino-5-amino-4-hydroxy-pentanoic acid, tert-butyl ester
hydroacetate (1.31 g, 2.9 mMol) in 15 mL of dichloromethane at
0.degree. C. was added benzenesulfonychloride (0.45 mL, 3.5 mMol)
followed by dropwise addition of N-methylmorpholine (0.8 mL, 9
mMol). The solution was left at room temperature for 16 hours, then
diluted with ethyl acetate (100 mL). The organic phase was washed
successively with sodium hydrogen carbonate and water, dried over
sodium sulfate, and concentrated under reduced pressure.
Chromatography over silica (elution with dichloromethane/methanol
15:1) gave 3-(1,2,3,4-tetrahydro-1-oxo-isoquinol-
ine-2-yl)-acetamino-5-benzenesulfonylamino-4-hydroxy-pentanoic
acid, tert-butyl ester (1.05 g).
[0204] Step G
[0205] To
3-(1,2,3,4-tetrahydro-1-oxo-isoquinoline-2-yl)-acetamino-5-benze-
nesulfonylamino-4-hydroxy-pentanoic acid, tert-butyl ester (0.9 g,
1.7 mMol) in 20 mL of dichloromethane was added 1.1.
1-triacetoxy-1.1-dihydro- -1.2-benziodoxol-3-1H)-one (Dess Martin
periodinane, 1.08 g, 2.5 mMol). After 2 hours at room temperature
the reaction mixture was diluted with ether, filtered, washed with
sodium hydrogen carbonate and water, dried over sodium sulfate, and
concentrated under reduced pressure. Chromatography over silica
(elution with dichloromethane/methanol 15:1) gave
3-(1,2,3,4-tetrahydro-1-oxo-isoquinoline-2-yl)-acetamino-5-benzenesu-
lfonylamino-4-oxo-pentanoic acid, tert-butyl ester (0.45 g).
[0206] Step H
[0207] A solution of
3-(1,2,3,4-tetrahydro-1-oxo-isoquinoline-2-yl)-acetam-
ino-5-benzenesulfonylamino-4-hydroxy-pentanoic acid, tert-butyl
ester (0.44 g, 0.48 mMol) and 15 mL of trifluoroacetic acid in 15
mL of dichloromethane was stirred at room temperature for 1 hour.
The solution was concentrated under reduced pressure.
Crystallization from dichloromethane/ether/hexane gave 0.17 g of
3-(1,2,3,4-tetrahydro-1-oxo-i-
soquinoline-2-yl)-acetamino-5-benzenesulfonylamino-4-oxo-pentanoic
acid.
[0208] The following compounds were also prepared according to the
procedure of Example 5 Steps F-H from the corresponding sulfonyl
chlorides:
EXAMPLE 5b
(S)-5-(Bicyclo[2.2.1]hept-1-ylmethanesulfonylamino)-4-oxo-3-[2-(1-oxo-3,4--
dihydro-1H-isoquinolin-2-yl)-acetylamino]-pentanoic acid
EXAMPLE 5c
(S)-4-Oxo-3-[2-(1-oxo-3,4-dihydro-1H-isoquinolin-2-yl)-acetylamino]-5-(2-p-
henyl-ethanesulfonylamino)-pentanoic acid
EXAMPLE 5d
4-Oxo-3-[2-(1-oxo-3,4-dihydro-1H-isoquinolin-2-yl)-acetylamino]-5-phenylme-
thanesulfonylamino-pentanoic acid
EXAMPLE 6a
3-[2-(2-Benzyloxycarbonylamino-4-tert-butoxycarbonyl-butyrylamino)-3-methy-
l-butyrylamino]-5-bromo-4-oxo-pentanoic acid tert-butyl ester
[0209] A solution of Z--Glu(OtBu)ValAsp(OtBu)--OH (14.9 g, 24.6
mmol) and 4-methylmorpholine (2.7 mL, 24.6 mmol) in 200 mL of THF
at ca. -40.degree. C. (dry ice CH.sub.3CN bath) was treated with
iso-butyl chloroformate (3.2 mL, 24.6 mmol). Solid immediately
formed. The sample was stirred for 15 minutes, then treated with
cold diazomethane (300 mL of an ether solution, freshly prepared
from Diazald). The sample was stirred at room temperature for 2
hours, cooled to 0.degree. C. and quenched by dropwise addition of
a 48% hydrobromic acid-acetic acid solution (35 mL of each). The
ice-bath was removed, the sample was stirred at room temperature
for 30 minutes, then extracted with ethyl acetate-water (500 mL of
each). The organic extract was washed with water, sat. NaHCO.sub.3
and brine solutions, dried (MgSO.sub.4), filtered and concentrated.
The residue was crystallized from dichloromethane-hexanes to give
10.5 g (63%) of 3-[2-(2-benzyloxy-carbony-
lamino-4-tert-butoxycarbonyl-butyrylamino)-3-methyl-butyrylamino]-5-bromo--
4-oxo-pentanoic acid tert-butyl ester
(Z--Glu(OtBu)ValAsp(OtBu)CH2Br) as a white solid.
[0210] Analysis Calculated for C.sub.31H.sub.46BrN.sub.3O.sub.9
(684.636): C, 54.39; H, 6.77; N, 6.14.
[0211] Found: C, 54.24; H, 6.63; N, 6.08.
[0212] Also prepared according to the procedure of Example 6a from
the corresponding peptides were:
EXAMPLE 6b
3-(2-Acetylamino-3-methyl-butyrylamino)-5-bromo-4-oxo-pentanoic
acid tert-butyl ester
EXAMPLE 6c
3-(2-Acetylamino-propylamino)-5-bromo-4-oxo-pentanoic acid
tert-butyl ester
EXAMPLE 6d
3-[2-(3-Phenyl-propionylamino-4-tert-butoxycarbonyl-butyrylamino)-3-methyl-
-butyrlamino]-5-bromo-4-oxo-pentanoic acid tert-butyl ester
EXAMPLE 6e
3-[2-(2-Benzyloxycarbonylamino-3-methyl-butyrylamino)-propionylamino]-5-br-
omo-4-oxo-pentanoic acid
[0213] Inhibition Studies
[0214] Compounds of Formula I and II are inhibitors of ICE as
demonstrated by measurement of K.sub.i (.mu.M) and IC.sub.50
(.mu.M) using the protocol described herein. ICE (0.24 nM final
concentration) is added to 400 .mu.L of HGDE buffer (100 mM HEPES,
20% glycerol, 5 mM DTT, 0.5 mM EDTA) containing 15 .mu.M substrate
(Ac-Tyr-Val-Ala-Asp-AMC; K.sub.M=15 .mu.M) plus vehicle (DMSO) or
inhibitor at concentrations bracketing the K.sub.i. Substrate
hydrolysis is monitored for 300 seconds by observing the
fluorescence of released AMC using excitation at 380 nm and
emission at 460 nm. Mean rates of substrate hydrolysis are
evaluated by linear-regression analysis of the fluorescence vs time
traces. To evaluate K.sub.i, plots of percent inhibition vs
inhibitor concentration are fit by non-linear regression to a
reversible, competitive model: 1 % Inhibition = 100 * [ I ] [ I ] +
K i * ( 1 + [ S ] K M )
[0215] Where the competition factor (1+[S]/K.sub.M)=2.
[0216] ICE Colorimetric Dose-Response (IC.sub.50) Assay
[0217] Diluted inhibitor stocks are prepared by two-fold serial
dilution from a primary stock whose concentration is selected
(based on screening results or on prior attempts at IC.sub.50
evaluation) to achieve approximately 95% inhibition in the most
concentrated well. Aliquots of each dilution are transferred to a
microtitre plate in triplicate.
[0218] ICE enzyme is diluted to approximately 24 nM in HGE buffer
(100 mM HEPES pH 7.5, 0.5 mM EDTA, 20% glycerol, 0.1% Bovine Serum
Albumin (BSA)), and activated by adding dithiothreitol (DTT) to a
final concentration of 5 mM. The activated enzyme is then aliquoted
into wells containing inhibitor or vehicle, and the plate is
preincubated for 60 minutes at ambient temperature. Substrate
(Ac-Tyr-Val-Ala-Asp-pNA) is added to each well to a final
concentration of 50 .mu.M, and plates are placed in the microtitre
plate-reader thermostated to 25.degree. C. Beginning 5 minutes
after addition of substrate, absorbance (405 nm) of wells is
monitored for 1 hour, and activity is calculated as the mean rate
of change in absorbance during this interval.
[0219] Ich-2 (Caspase-4) Colorimetric Dose-Response (IC.sub.50)
Assay
[0220] Inhibition of Ich-2 enzyme is assayed as described above for
ICE, except that enzyme is used at 64 nM, and 60 gM of the
Ich-2-specific substrate Ac-Leu-Glu-Val-Asp-pNA is used instead of
the ICE substrate Ac-Tyr-Val-Ala-Asp-pNA.
[0221] The results of these assays are shown below in Table 1.
1 TABLE 1 Ich-2 IC.sub.50 ICE Ki ICE IC.sub.50 (Caspase-4) Example
Number (.mu.M) (.mu.M) (.mu.M) 2a 11 73 96 2b 32 245 -- 2c 14 168
124 2a 11.0 73.0 445.0 2b 34.5 245.0 8591.0 2c 18.0 168.0 2339.0 2d
37.0 291.0 2e 735.0 1833.0 2f 22.0 174.0 1008.0 2g 33.0 136.0 314.0
2h 16.0 55.6 199.0 2i 30.1 25.0 59.0 2j 65.0 194.0 159.0 2k 22.0 2l
1.4 28.0 935.0 2m 2.8 35.3 3a 0.007 0.072 18.0 3b 0.013 0.025 4.2
3c 0.0051 0.009 3.4 3d 0.0078 0.003 0.9 4a 0.27 7.6 4c 0.001 0.015
8.5 4d 0.0016 0.003 1.6 4e 0.00011 0.002 0.7 4f 0.00061 0.002 3.2
5a 105.0 586.0 5b 20.0 165.0 252.0 5c 45.0 371.0 5d 27.0 234.0
HEPES = 4-(2-hydroxymethyl)-1-piperazine ethane sulfonic acid DTT =
Dithiothreitol EDTA = Ethylene diamine tetra acetic acid AMC =
7-amino-4-methyl coumarin Tyr = Tyrosine Val = Valine Ala = Alanine
Asp = Aspartic Acid pNA = Para nitroaniline LEU = Leucine Glu =
Glutamic acid Me = Methyl t-Bu = Tert butyl
[0222] Calculated for
C.sub.29H.sub.38N.sub.4O.sub.9S.sub.1-0.33CF.sub.3CO- OH: C, 54,26;
H, 5.88; N, 8.53.
[0223] Found: Cm 54,26; H, 5.93; N, 8.55.
EXAMPLE 3b
[0224]
3[2-(2-Benzyloxycarbonylamino-4-carboxy-butyrylarnino)-3-methyl-
bytyrylamino]-4-oxo-5-(2-phenyl-ethanesulfonylamino)-paentanoic
acid
EXAPLE 3c
[0225]
3-{2-[4-Carboxy-2-(3-phenyl-propionylamino)-butyrylamino]-3-methyl--
butyrylamino}-4-oxo-5-(2-phenyl-ehtanesulfonylamino)-pentanoic
acid
[0226] Calculated for
C.sub.32H.sub.42N.sub.4O.sub.10S.sub.1.cndot.0.24CF.- sub.3COOH: C,
55.56; H, 6.06; N, 7.98.
[0227] Found C, 55.56; Hm 6.23; N, 8.07.
EXAMPLE 3d
[0228]
3-(2-{2-[2-Acetylamino-3-(4-hydroxy-phenyl)-propionylamino]-4-carbo-
xy-butyrylamino}-3-methyl -abutyrylamino)-4-oxo-5-(2-jemu;
-ethanesulfonylamino)-pentanoic acid
[0229] The following compounds were prepared according tho the
procedure of Exaple 2a from the corresponding protected di, tro or
terrapeptide bromonethylketones and
tert-butyl[7,7-dimethyl-2-oxo-bucyclo[2.2.1]hept-1-
-ylmethanesulfonyl ]carbamate:
EXAMPLE 4a
[0230]
3-(2-Acetylamino-3-methyl-butyrylamino)-5-(7,7-dimehtyl-2-oxo-bicyc-
lo[2.2.1]hept-1-(S)-ylmethanesulfonylamino)-4-oxo-pentanoic
acid
[0231] MS (AP+): 502.
[0232] IR (KBr,cm.sup.-1): 3338,2965,1738,1653,1540,
1395,1328,1148.
Sequence CWU 1
1
6 1 4 PRT Artificial Sequence UNSURE (4)
(2-phenyl-ethanesulfonylamino) attached to 3' end 1 Tyr Glu Val Asp
1 2 4 PRT Artificial Sequence UNSURE (4)
(7,7-dimethyl-2-oxobycyclo[2.2.1]hept-1- ylmethanesuylfonylamino)
attached to 3'end 2 Tyr Glu Val Asp 1 3 4 PRT Artificial Sequence
UNSURE (1)..(4) Acetyl group attached to 5' end and AMC group
attached to 3' end 3 Tyr Val Ala Asp 1 4 4 PRT Artificial Sequence
UNSURE (1)..(4) Acetyl group attached to 5' end and pNA group
attached to 3'end 4 Tyr Val Ala Asp 1 5 4 PRT Artificial Sequence
UNSURE (1)..(4) Acetyl group attached to 5' end and pNA group
attached to 3' end 5 Leu Glu Val Asp 1 6 4 PRT Artificial Sequence
UNSURE (1)..(4) Acetyl group attached to 5' end and pNA group
attached to 3' end 6 Tyr Val Ala Asp 1
* * * * *