U.S. patent application number 11/273850 was filed with the patent office on 2006-03-30 for peptide-containing alpha-ketoamide cysteine and serine protease inhibitors.
This patent application is currently assigned to Cephalon, Inc.. Invention is credited to Ron Bihovsky, Sankar Chatterjee, John P. Mallamo, Gregory J. Wells.
Application Number | 20060069037 11/273850 |
Document ID | / |
Family ID | 26740934 |
Filed Date | 2006-03-30 |
United States Patent
Application |
20060069037 |
Kind Code |
A1 |
Chatterjee; Sankar ; et
al. |
March 30, 2006 |
Peptide-containing alpha-ketoamide cysteine and serine protease
inhibitors
Abstract
This invention relates to peptide-containing .alpha.-ketoamide
inhibitors of cysteine and serine proteases, methods for making
these compounds, and methods for using the same.
Inventors: |
Chatterjee; Sankar;
(Wynnewood, PA) ; Mallamo; John P.; (Glenmoore,
PA) ; Bihovsky; Ron; (Wynnewood, PA) ; Wells;
Gregory J.; (West Chester, PA) |
Correspondence
Address: |
CEPHALON, INC.
41 MOORES ROAD
PO BOX 4011
FRAZER
PA
19355
US
|
Assignee: |
Cephalon, Inc.
Frazer
PA
|
Family ID: |
26740934 |
Appl. No.: |
11/273850 |
Filed: |
November 15, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10685923 |
Oct 14, 2003 |
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11273850 |
Nov 15, 2005 |
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09879336 |
Jun 12, 2001 |
6703368 |
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10685923 |
Oct 14, 2003 |
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09527540 |
Mar 16, 2000 |
6288231 |
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09879336 |
Jun 12, 2001 |
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09166808 |
Oct 6, 1998 |
6150378 |
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09527540 |
Mar 16, 2000 |
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60061309 |
Oct 7, 1997 |
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Current U.S.
Class: |
514/252.12 ;
514/20.3; 530/330; 530/331 |
Current CPC
Class: |
A61P 11/00 20180101;
C07D 211/96 20130101; A61P 25/08 20180101; C07D 409/14 20130101;
A61P 25/16 20180101; C07K 5/0202 20130101; C07D 213/74 20130101;
A61P 9/10 20180101; A61P 9/12 20180101; C07D 275/06 20130101; C07D
333/62 20130101; A61P 19/02 20180101; C07D 213/70 20130101; A61P
43/00 20180101; C07C 311/46 20130101; C07D 211/76 20130101; C07D
207/08 20130101; C07D 235/06 20130101; C07D 239/42 20130101; C07C
311/48 20130101; A61P 25/28 20180101; C07D 333/34 20130101; C07D
207/20 20130101; C07D 207/48 20130101; C07D 277/40 20130101; C07K
5/06069 20130101; C07D 277/54 20130101; A61P 11/06 20180101; C07D
413/04 20130101; C07C 311/06 20130101; C07D 409/04 20130101; A61P
7/02 20180101; A61P 1/18 20180101 |
Class at
Publication: |
514/018 ;
514/252.12; 514/019; 530/330; 530/331 |
International
Class: |
A61K 38/05 20060101
A61K038/05; A61K 38/04 20060101 A61K038/04; C07K 5/06 20060101
C07K005/06; C07K 5/04 20060101 C07K005/04 |
Claims
1. A compound having the Formula I: ##STR78## wherein: Q has the
formula G-B-(CHR.sup.4).sub.v where R.sup.4 is independently H or
alkyl having from 1 to 4 carbons; v is 0; B is selected from the
group consisting of --C(.dbd.O)--, --OC(.dbd.O)--, --S--, --SO--,
--S(O).sub.2-- and a bond; M is a carbon atom; G is selected from
the group consisting of H, a blocking group, lower alkyl, lower
alkenyl, aryl having from about 6 to about 14 carbons, and
arylalkyl having from about 7 to about 15 carbons, said alkyl and
arylalkyl groups being optionally substituted with one or more J
groups; J is selected from the group consisting of halogen, CN,
nitro, lower alkyl, cycloalkyl, heterocycloalkyl, heteroalkyl,
halogenated alkyl, aryloxyalkyl, alkylthio, alkylsulfonyl, aryl,
heteroaryl, arylalkyl, arylalkyloxy, arylsulfonyl,
heteroarylsulfonyl, alkoxycarbonyl, alkoxyalkyl, acyl, alkoxy,
hydroxy, carboxy, hydroxyalkyl, amino, alkylamino, and aminoalkyl,
said amino group or said amino group of said aminoalkyl or
alkylamino group being optionally substituted with an acyl group,
an alkoxy group, or with 1 to 3 aryl, lower alkyl, cycloalkyl, or
alkoxyalkyl groups; and said aryl, heteroaryl, heterocycloalkyl,
and heteroalkyl groups being further optionally substituted by a
J.sup.1 group; J.sup.1 is selected from the group consisting of
halogen, CN, nitro, lower alkyl, cycloalkyl, heterocycloalkyl,
heteroalkyl, halogenated alkyl, aryloxyalkyl, alkylthio,
alkylsulfonyl, aryl, heteroaryl, arylalkyl, arylalkyloxy,
arylsulfonyl, heteroarylsulfonyl, alkoxycarbonyl, alkoxyalkyl,
acyl, alkoxy, hydroxy, carboxy, hydroxyalkyl, amino, alkylamino,
and aminoalkyl, said amino group or said amino group of said
aminoalkyl or alkylamino group being optionally substituted with an
acyl group, an alkoxy group, or with 1 to 3 aryl, lower alkyl,
cycloalkyl, or alkoxyalkyl groups; and said aryl, heteroaryl,
heterocycloalkyl, and heteroalkyl groups being further optionally
substituted by a J.sup.2 group; J.sup.2 is selected from a group
consisting of halogen, CN, nitro, lower alkyl, halogenated alkyl,
alkylthio, alkylsulfonyl, alkoxycarbonyl, alkoxyalkyl, acyl,
alkoxy, hydroxy, carboxy, hydroxyalkyl, amino, alkylamino, and
aminoalkyl; each Aaa is independently an amino acid; n is 0 or 1;
R.sup.1 is selected from the group consisting of H, alkyl having
from one to about 6 carbons, arylalkyl having from about 7 to about
15 carbons, alkoxyalkyl, and a side chain of a naturally occurring
amino acid in the R or S configuration, said alkyl, arylalkyl, and
alkoxyalkyl groups being optionally substituted with one or more J
groups; R.sup.2 is benzyl; R.sup.3 is selected from the group
consisting of H, alkyl having from one to about 6 carbons,
arylalkyl having from about 7 to about 15 carbons, alkoxyalkyl, a
side chain of a naturally occurring amino acid in the R or S
configuration, C(.dbd.O)R.sup.7, S(.dbd.O).sub.2R.sup.7, a blocking
group, and said alkyl, arylalkyl, and alkoxyalkyl groups being
optionally substituted with one or more J groups; R.sup.7 is
selected from the group consisting of aryl having from about 6 to
about 14 carbons, heteroaryl having from about 5 to about 14 ring
atoms, arylalkyl having from about 7 to about 15 carbons, alkyl
having from 1 to about 10 carbons, said aryl, heteroaryl, arylalkyl
and alkyl groups being optionally substituted with one or more J
groups, heteroalkyl having from 2 to about 7 carbons, alkoxy having
from about 1 to about 10 carbons, and amino optionally substituted
with 1 or more alkyl groups; q is 0 or 1; Z is
C(.dbd.O)C(.dbd.O)NH--X-A.sup.1-K; X is a bond; A.sup.1 is lower
alkylene; K is N(R.sup.10)SO.sub.2R.sup.8 or
SO.sub.2N(R.sup.8)(R.sup.10); R.sup.8 is thiophene, said thiophene
being substituted with one or more J groups; R.sup.10 is selected
from the group consisting of H and lower alkyl; or a
pharmaceutically acceptable salt thereof.
2. The compound of claim 1 wherein n is 0, q is 1, B is a bond, and
G is H.
3. The compound of claim 1 wherein R.sup.1 is the sidechain of a
naturally occurring amino acid.
4. The compound of claim 1 wherein R.sup.3 is
--S(.dbd.O).sub.2R.sup.7.
5. The compound of claim 1 wherein A.sup.1 is
--CH.sub.2--CH.sub.2--, --CH.sub.2--CH(CH.sub.3)-- or
--CH.sub.3(CH)--CH.sub.2--.
6. The compound of claim 1 wherein R.sup.1 is a serine sidechain,
which is optionally capped with a benzyl group.
7. The compound of claim 6 wherein M is a carbon atom in the D
configuration.
8. The compound of claim 1 wherein R.sup.7 is methyl.
9. The compound of claim 1 wherein n is 0, q is 1, R.sup.1 is the
side chain of an amino acid in the D- or L-configuration, R.sup.3
is S(.dbd.O).sub.2R.sup.7, G is H and B is a bond.
10. The compound of claim 1 wherein R.sup.1 is a serine side chain
in the D-configuration in which the hydroxyl group is capped with
benzyl and R.sup.7 is methyl.
11. The compound of claim 1 wherein n and q are each 0; B is
(C.dbd.O); and G is phenyl or lower alkyl, said phenyl or lower
alkyl groups being optionally substituted with one or more J
groups.
12. A composition for inhibiting a serine protease or a cysteine
protease comprising a compound of claim 1 and a pharmaceutically
acceptable carrier.
13. A method for inhibiting a serine protease or a cysteine
protease comprising contacting a protease selected from the group
consisting of serine proteases and cysteine proteases with an
inhibitory amount of a compound of claim 1.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional of U.S. Ser. No. 10/685,923
filed Oct. 14, 2003, now allowed; which is a divisional of U.S.
Ser. No. 09/879,336 filed Jun. 21, 2001, now U.S. Pat. No.
6,703,368; which is a divisional of U.S. Ser. No. 09/527,540 filed
Mar. 16, 2000, now U.S. Pat. No. 6,288,231; which is a divisional
of U.S. Ser. No. 09/166,808 filed Oct. 6, 1998, now U.S. Pat. No.
6,150,378; and claims benefit of U.S. Provisional Application Ser.
No. 60/061,309, filed Oct. 7, 1997, the disclosure of which is
hereby incorporated herein by reference in its entirety.
FIELD OF THE INVENTION
[0002] This invention relates to peptide-containing
.alpha.-ketoamide inhibitors of cysteine and serine proteases,
methods for making these compounds, and methods for using the
same.
BACKGROUND OF THE INVENTION
[0003] Numerous cysteine and serine proteases have been identified
in human tissues. A "protease" is an enzyme which degrades proteins
into smaller components (peptides). The terms "cysteine protease"
and "serine protease" refer to proteases which are distinguished by
the presence therein of a cysteine or serine residue which plays a
critical role in the catalytic process. Mammalian systems,
including humans, normally degrade and process proteins via a
variety of enzymes including cysteine and serine proteases.
However, when present at elevated levels or when abnormally
activated, cysteine and serine proteases may be involved in
pathophysiological processes.
[0004] For example, calcium-activated neutral proteases
("calpains") comprise a family of intracellular cysteine proteases
which are ubiquitously expressed in mammalian tissues. Two major
calpains have been identified; calpain I and calpain II. While
calpain II is the predominant form in many tissues, calpain I is
thought to be the predominant form in pathological conditions of
nerve tissues. The calpain family of cysteine proteases has been
implicated in many diseases and disorders, including
neurodegeneration, stroke, Alzheimer's, amyotrophy, motor neuron
damage, acute central nervous system injury, muscular dystrophy,
bone resorption, platelet aggregation, cataracts and inflammation.
Calpain I has been implicated in excitatory amino-acid induced
neurotoxicity disorders including ischemia, hypoglycemia,
Huntington's Disease, and epilepsy. The lysosomal cysteine protease
cathepsin B has been implicated in the following disorders:
arthritis, inflammation, myocardial infarction, tumor metastasis,
and muscular dystrophy. Other lysosomal cysteine proteases include
cathepsins C, H, L and S. Interleukin-1.beta. converting enzyme
("ICE") is a cysteine protease which catalyzes the formation of
interleukin-1.beta.. Interleukin-1.beta. is an immunoregulatory
protein implicated in the following disorders: inflammation,
diabetes, septic shock, rheumatoid arthritis, and Alzheimer's
disease. ICE has also been linked to apoptotic cell death of
neurons, which is implicated in a variety of neurodegenerative
disorders including Parkinson's disease, ischemia, and amyotrophic
lateral sclerosis (ALS).
[0005] Cysteine proteases are also produced by various pathogens.
The cysteine protease clostripain is produced by Clostridium
histolyticum. Other proteases are produced by Trypanosoma cruzi,
malaria parasites Plasmodium falciparum and P. vinckei and
Streptococcus. Hepatitis A viral protease HAV C3 is a cysteine
protease essential for processing of picornavirus structural
proteins and enzymes.
[0006] Exemplary serine proteases implicated in degenerative
disorders include thrombin, human leukocyte elastase, pancreatic
elastase, chymase and cathepsin G. Specifically, thrombin is
produced in the blood coagulation cascade, cleaves fibrinogen to
form fibrin and activates Factor VIII; thrombin is implicated in
thrombophlebitis, thrombosis and asthma. Human leukocyte elastase
is implicated in tissue degenerative disorders such as rheumatoid
arthritis, osteoarthritis, atherosclerosis, bronchitis, cystic
fibrosis, and emphysema. Pancreatic elastase is implicated in
pancreatitis. Chymase, an enzyme important in angiotensin
synthesis, is implicated in hypertension, myocardial infarction,
and coronary heart disease. Cathepsin G is implicated in abnormal
connective tissue degradation, particularly in the lung.
[0007] Given the link between cysteine and serine proteases and
various debilitating disorders, compounds which inhibit these
proteases would be useful and would provide an advance in both
research and clinical medicine. The present invention is directed
to these, as well as other, important ends.
SUMMARY OF THE INVENTION
[0008] The present invention is directed to selected
peptide-containing .alpha.-ketoamide inhibitors of cysteine and
serine proteases represented by the general formula I: ##STR1##
wherein: [0009] Q has the formula G-B-(CHR.sup.4).sub.v where
R.sup.4 is independently H or alkyl having from 1 to 4 carbons;
[0010] v is 0, 1, or 2; [0011] B is selected from the group
consisting of C(.dbd.O), OC(.dbd.O), S(.dbd.O).sub.m, CH.sub.2, a
bond, NR.sup.5C(.dbd.O), S(.dbd.O).sub.m-A-C(.dbd.O), and
C(.dbd.O)-A-C(.dbd.O), where R.sup.5 is H or lower alkyl; [0012] m
is 0, 1, or 2; [0013] A is lower alkylene or cycloalkylene,
optionally substituted with one or more halogen atoms, aryl, or
heteroaryl groups; [0014] M is a carbon atom; [0015] G is selected
from the group consisting of H, a blocking group, lower alkyl,
lower alkenyl, aryl having from about 6 to about 14 carbons,
heterocyclyl having from about 5 to about 14 ring atoms,
heterocycloalkyl having from about 5 to about 14 ring atoms,
arylalkyl having from about 7 to about 15 carbons, heteroarylalkyl,
and arylheteroalkyl wherein the aryl portion can be unfused or
fused with the heteroalkyl ring, said alkyl, aryl, heterocyclyl,
heterocycloalkyl, arylalkyl, heteroarylalkyl, and arylheteroalkyl
groups being optionally substituted with one or more J groups;
[0016] J is selected from the group consisting of halogen, CN,
nitro, lower alkyl, cycloalkyl, heterocycloalkyl, heteroalkyl,
halogenated alkyl, aryloxyalkyl, alkylthio, alkylsulfonyl, aryl,
heteroaryl, arylalkyl, arylalkyloxy, arylsulfonyl,
heteroarylsulfonyl, alkoxycarbonyl, alkoxyalkyl, acyl, alkoxy,
hydroxy, carboxy, hydroxyalkyl, amino, alkylamino, and aminoalkyl,
said amino group or said amino group of said aminoalkyl or
alkylamino group being optionally substituted with an acyl group,
an alkoxy group, or with 1 to 3 aryl, lower alkyl, cycloalkyl, or
alkoxyalkyl groups; and said aryl, heteroaryl, heterocycloalkyl,
and heteroalkyl groups being further optionally substituted by a J
group; [0017] each Aaa is independently an amino acid which
optionally contains one or more blocking groups; [0018] n is 0, 1,
2, or 3; [0019] R.sup.1 and R.sup.2 are independently selected from
the group consisting of H, alkyl having from one to about 6
carbons, arylalkyl having from about 7 to about 15 carbons,
heteroalkyl in which the ring contains from about 5 to about 14
ring atoms, heteroarylalkyl in which the heteroaryl ring contains
from about 5 to about 14 ring atoms, alkoxyalkyl, a side chain of a
naturally occurring amino acid in the R or S configuration, and
(CH.sub.2).sub.pNH-L, said alkyl, arylalkyl, heteroalkyl,
heteroarylalkyl, and alkoxyalkyl groups being optionally
substituted with one or more J groups; [0020] p is 0, 1, 2, or 3;
[0021] L is selected from the group consisting of alkoxycarbonyl
having from 2 to about 7 carbons, arylalkoxycarbonyl in which the
arylalkoxy group contains about 7 to about 15 carbons, and
S(.dbd.O).sub.2R.sup.6; [0022] R.sup.6 is selected from the group
consisting of lower alkyl, and aryl having from about 6 to about 14
carbons; [0023] R.sup.3 is selected from the group consisting of H,
alkyl having from one to about 6 carbons, arylalkyl having from
about 7 to about 15 carbons, heteroalkyl in which the ring contains
from about 5 to about 14 ring atoms, heteroarylalkyl in which the
heteroaryl ring contains from about 5 to about 14 ring atoms,
alkoxyalkyl, a side chain of a naturally occurring amino acid in
the R or S configuration, (CH.sub.2).sub.pNH-L, C(.dbd.O)R.sup.7,
S(.dbd.O).sub.2R.sup.7, a blocking group, and when combined with
the carbon atom to which R.sup.1 is attached an alkylene group
having from 2 to 5 carbons, said alkylene group being optionally
substituted with a group selected from the group consisting of
aryl, azide, CN, a protected amino group, and OSO.sub.2-aryl, said
alkyl, arylalkyl, heteroalkyl, heteroarylalkyl, and alkoxyalkyl
groups being optionally substituted with one or more J groups;
[0024] R.sup.7 is selected from the group consisting of aryl having
from about 6 to about 14 carbons, heteroaryl having from about 5 to
about 14 ring atoms, arylalkyl having from about 7 to about 15
carbons, alkyl having from 1 to about 10 carbons, said aryl,
heteroaryl, arylalkyl and alkyl groups being optionally substituted
with one or more J groups, heteroalkyl having from 2 to about 7
carbons, alkoxy having from about 1 to about 10 carbons, and amino
optionally substituted with 1 or more alkyl groups; [0025] q is 0
or 1; [0026] Z is selected from the group consisting of
C(.dbd.O)C(.dbd.O)NH--X-A.sup.1-K and ##STR2## [0027] X is a bond
or --O--; [0028] A.sup.1 is the same as A; [0029] K is selected
from the group consisting of N(R.sup.10)Y, ##STR3## [0030] D is a
fused aryl or heteroaryl group; [0031] R.sup.11 is selected from
the group consising of alkoxy, aryloxy, and NHR.sup.12; [0032]
R.sup.12 is selected from the group consisting of H, alkyl, aryl,
and heteroaryl, said alkyl, aryl or heteroaryl groups being
optionally substituted with one or more J groups; [0033] Y is
selected from the group consisting of SO.sub.2R.sup.8,
C(.dbd.O)NHR.sup.9, C(.dbd.S)NHR.sup.9, C(.dbd.NCN)R.sup.11,
C(.dbd.NC(.dbd.O)NHR.sup.10)R.sup.11, and CO.sub.2R.sup.8; [0034]
R.sup.8 is selected from the group consisting of alkyl, alkoxy,
aryl, and heterocyclyl, said alkyl, alkoxy, aryl, or heterocyclyl
groups being optionally substituted with one or more J groups;
[0035] R.sup.9 is selected from the group consisting of H, alkyl,
aryl, and heteroaryl, said alkyl, aryl, or heteroaryl groups being
optionally substituted with one or more J groups; [0036] or an
R.sup.9 alkyl group may be combined with an A.sup.1 alkylene group
to form a N-containing heterocyclic 5- or 6-membered ring; [0037]
R.sup.10 is selected from the group consisting of H and lower
alkyl; [0038] or in the moiety SO.sub.2N(R.sup.8)R.sup.10, R.sup.8
and R.sup.10 may be combined together with the N atom to which they
are attached to form a N-containing heterocyclic 5- or 6-membered
ring; [0039] or where A.sup.1 is an alkylene group, and K is
N(R.sup.10)Y wherein R.sup.10 is alkyl, said R.sup.10 alkyl group
may be combined with said A.sup.1 alkylene group to form a
N-containing heterocyclic 5- or 6-membered ring; or a
pharmaceutically acceptable salt thereof.
[0040] In some preferred embodiments of the compounds of Formula I,
n and v are each 0, q is 1, B is a bond, and G is H. In further
preferred embodiments of the compounds of Formula I, R.sup.1 is the
sidechain of a naturally occurring amino acid. In still further
preferred embodiments of the compounds of Formula I, R.sup.3 is
--S(.dbd.O).sub.2R.sup.7.
[0041] In some preferred embodiments of the compounds of Formula I,
R.sup.2 is benzyl or alkoxyalkyl. In more preferred embodiments, X
is a bond, and Y is SO.sub.2R.sup.8. Preferably, A.sup.1 is
--CH.sub.2--CH.sub.2--, --CH.sub.2--CH(CH.sub.3)--, or
--(CH.sub.3)CH--CH.sub.2--.
[0042] In further preferred embodiments of the compounds of Formula
I, R.sup.1 is a serine sidechain, which is optionally capped with a
benzyl group. Preferably, the carbon to which the serine sidechain
is attached, designated "M" in Formula I, is a carbon atom in the D
configuration.
[0043] In preferred embodiments of the compounds of Formula I,
R.sup.2 is benzyl, R.sup.7 is methyl, and R.sup.8 is substituted
phenyl, unsubstituted phenyl, substituted heteroaryl, or
unsubstituted heteroaryl. In particularly preferred embodiments,
R.sup.8 is aryl, aryl substituted with amino, aryl substituted with
heterocyclomethyl, heteroaryl, alkyl substituted with heteroaryl,
or heteroaryl substituted with alkylthio, haloalkyl, alkyl,
phenylsulfonyl, halogen, aminophenyl, amino, or
dialkylaminoalkyl.
[0044] In further preferred embodiments of the compounds of Formula
I, n, v and q are each 0, B is (C.dbd.O), and G is phenyl or lower
alkyl, said phenyl or lower alkyl groups being optionally
substituted with one or more J groups.
[0045] In more preferred embodiments of the invention, n and v are
each 0, q is 1, R.sup.1 is the side chain of an amino acid in the
D- or L-configuration, R.sup.3 is S(.dbd.O).sub.2R.sup.7, G is H, B
is a bond, R.sup.2 is benzyl or alkoxyalkyl, X is a bond, and Y is
SO.sub.2R.sup.8.
[0046] In other preferred embodiments, A.sup.1 is CH.sub.2CH.sub.2,
CH.sub.2CH(CH.sub.3), or (CH.sub.3)CHCH.sub.2. In more preferred
embodiments, R.sup.1 is a serine side chain in the D-configuration
in which the hydroxyl group is capped with benzyl, R.sup.2 is
benzyl, R.sup.7 is methyl, and R.sup.8 is substituted or
unsubstituted phenyl or substituted or unsubstituted
heteroaryl.
[0047] More preferred are the substituents shown for
R.sup.1-R.sup.4, B, G, Aaa, X, A.sup.1, Y, n, q and v shown for the
compounds in Tables 2, 3, 4 and 5. Especially preferred are the
substituents shown for compounds 9, 13, 17, 22, and 29-151.
[0048] Some especially preferred embodiments of the compounds of
Formula I are shown in Tables 2, 3, 4 and 5, infra, with compounds
9, 13, 17, 22, and 29-151 being particularly preferred.
[0049] Because the peptide-containing .alpha.-ketoamides of the
invention inhibit cysteine proteases and serine proteases, they can
be used in both research and therapeutic settings.
[0050] In a research environment, preferred compounds having
defined attributes can be used to screen for natural and synthetic
compounds which evidence similar characteristics in inhibiting
protease activity. The compounds can also be used in the refinement
of in vitro and in vivo models for determining the effects of
inhibition of particular proteases on particular cell types or
biological conditions.
[0051] In a therapeutic setting, given the connection between
cysteine proteases and certain defined disorders, and serine
proteases and certain defined disorders, compounds of the invention
can be utilized to alleviate, mediate, reduce and/or prevent
disorders which are associated with abnormal and/or aberrant
activity of cysteine proteases and/or serine proteases.
[0052] In preferred embodiments, compositions are provided for
inhibiting a serine protease or a cysteine protease comprising a
compound of the invention and a pharmaceutically acceptable
carrier. In other preferred embodiments, methods are provided for
inhibiting serine proteases or cysteine proteases comprising
contacting a protease selected from the group consisting of serine
proteases and cysteine proteases with an inhibitory amount of a
compound of the invention.
[0053] Methodologies for making the present peptide-containing
.alpha.-ketoamide inhibitors are also disclosed. Other useful
methodologies will be apparent to those skilled in the art, once
armed with the present disclosure. These and other features of the
compounds of the subject invention are set forth in more detail
below.
DETAILED DESCRIPTION
[0054] Disclosed herein are the selected peptide-containing
.alpha.-ketoamides which are represented by the following formula
I: ##STR4## wherein: [0055] Q has the formula G-B-(CHR.sup.4).sub.v
where R.sup.4 is independently H or alkyl having from 1 to 4
carbons; [0056] v is 0, 1, or 2; [0057] B is selected from the
group consisting of C(.dbd.O), OC(.dbd.O), S(.dbd.O).sub.m,
CH.sub.2, a bond, NR.sup.5C(.dbd.O), S(.dbd.O).sub.m-A-C(.dbd.O),
and C(.dbd.O)-A-C(.dbd.O), where R.sup.5 is H or lower alkyl;
[0058] m is 0, 1, or 2; [0059] A is lower alkylene or
cycloalkylene, optionally substituted with one or more halogen
atoms, aryl, or heteroaryl groups; [0060] M is a carbon atom;
[0061] G is selected from the group consisting of H, a blocking
group, lower alkyl, lower alkenyl, aryl having from about 6 to
about 14 carbons, heterocyclyl having from about 5 to about 14 ring
atoms, heterocycloalkyl having from about 5 to about 14 ring atoms,
arylalkyl having from about 7 to about 15 carbons, heteroarylalkyl,
and arylheteroalkyl wherein the aryl portion can be unfused or
fused with the heteroalkyl ring, said alkyl, aryl, heterocyclyl,
heterocycloalkyl, arylalkyl, heteroarylalkyl, and arylheteroalkyl
groups being optionally substituted with one or more J groups;
[0062] J is selected from the group consisting of halogen, CN,
nitro, lower alkyl, cycloalkyl, heterocycloalkyl, heteroalkyl,
halogenated alkyl, aryloxyalkyl, alkylthio, alkylsulfonyl, aryl,
heteroaryl, arylalkyl, arylalkyloxy, arylsulfonyl,
heteroarylsulfonyl, alkoxycarbonyl, alkoxyalkyl, acyl, alkoxy,
hydroxy, carboxy, hydroxyalkyl, amino, alkylamino, and aminoalkyl,
said amino group or said amino group of said aminoalkyl or
alkylamino group being optionally substituted with an acyl group,
an alkoxy group, or with 1 to 3 aryl, lower alkyl, cycloalkyl, or
alkoxyalkyl groups; and said aryl, heteroaryl, heterocycloalkyl,
and heteroalkyl groups being further optionally substituted by a J
group; [0063] each Aaa is independently an amino acid which
optionally contains one or more blocking groups; [0064] n is 0, 1,
2, or 3; [0065] R.sup.1 and R.sup.2 are independently selected from
the group consisting of H, alkyl having from one to about 6
carbons, arylalkyl having from about 7 to about 15 carbons,
heteroalkyl in which the ring contains from about 5 to about 14
ring atoms, heteroarylalkyl in which the heteroaryl ring contains
from about 5 to about 14 ring atoms, alkoxyalkyl, a side chain of a
naturally occurring amino acid in the R or S configuration, and
(CH.sub.2).sub.pNH-L, said alkyl, arylalkyl, heteroalkyl,
heteroarylalkyl, and alkoxyalkyl groups being optionally
substituted with one or more J groups; [0066] p is 0, 1, 2, or 3;
[0067] L is selected from the group consisting of alkoxycarbonyl
having from 2 to about 7 carbons, arylalkoxycarbonyl in which the
arylalkoxy group contains about 7 to about 15 carbons, and
S(.dbd.O).sub.2R.sup.6; [0068] R.sup.6 is selected from the group
consisting of lower alkyl, and aryl having from about 6 to about 14
carbons; [0069] R.sup.3 is selected from the group consisting of H,
alkyl having from one to about 6 carbons, arylalkyl having from
about 7 to about 15 carbons, heteroalkyl in which the ring contains
from about 5 to about 14 ring atoms, heteroarylalkyl in which the
heteroaryl ring contains from about 5 to about 14 ring atoms,
alkoxyalkyl, a side chain of a naturally occurring amino acid in
the R or S configuration, (CH.sub.2).sub.pNH-L, C(.dbd.O)R.sup.7,
S(.dbd.O).sub.2R.sup.7, a blocking group, and when combined with
the carbon atom to which R.sup.1 is attached an alkylene group
having from 2 to 5 carbons, said alkylene group being optionally
substituted with a group selected from the group consisting of
aryl, azide, CN, a protected amino group, and OSO.sub.2-aryl, said
alkyl, arylalkyl, heteroalkyl, heteroarylalkyl, and alkoxyalkyl
groups being optionally substituted with one or more J groups;
[0070] R.sup.7 is selected from the group consisting of aryl having
from about 6 to about 14 carbons, heteroaryl having from about 5 to
about 14 ring atoms, arylalkyl having from about 7 to about 15
carbons, alkyl having from 1 to about 10 carbons, said aryl,
heteroaryl, arylalkyl and alkyl groups being optionally substituted
with one or more J groups, heteroalkyl having from 2 to about 7
carbons, alkoxy having from about 1 to about 10 carbons, and amino
optionally substituted with 1 or more alkyl groups; [0071] q is 0
or 1; [0072] Z is selected from the group consisting of
C(.dbd.O)C(.dbd.O)NH--X-A.sup.1-K and ##STR5## [0073] X is a bond
or --O--; [0074] A.sup.1 is the same as A; [0075] K is selected
from the group consisting of N(R.sup.10)Y, ##STR6## [0076] D is a
fused aryl or heteroaryl group; [0077] R.sup.11 is selected from
the group consising of alkoxy, aryloxy, and NHR.sup.12; [0078]
R.sup.12 is selected from the group consisting of H, alkyl, aryl,
and heteroaryl, said alkyl, aryl or heteroaryl groups being
optionally substituted with one or more J groups; [0079] Y is
selected from the group consisting of SO.sub.2R.sup.8,
C(.dbd.O)NHR.sup.9, C(.dbd.S)NHR.sup.9, C(.dbd.NCN)R.sup.11,
C(.dbd.NC(.dbd.O)NHR.sup.10)R.sup.11, and CO.sub.2R.sup.8; [0080]
R.sup.8 is selected from the group consisting of alkyl, alkoxy,
aryl, and heterocyclyl, said alkyl, alkoxy, aryl, or heterocyclyl
groups being optionally substituted with one or more J groups;
[0081] R.sup.9 is selected from the group consisting of H, alkyl,
aryl, and heteroaryl, said alkyl, aryl, or heteroaryl groups being
optionally substituted with one or more J groups; [0082] or an
R.sup.9 alkyl group may be combined with an A.sup.1 alkylene group
to form a N-containing heterocyclic 5- or 6-membered ring; [0083]
R.sup.10 is selected from the group consisting of H and lower
alkyl; [0084] or in the moiety SO.sub.2N(R.sup.8)R.sup.10, R.sup.8
and R.sup.10 may be combined together with the N atom to which they
are attached to form a N-containing heterocyclic 5- or 6-membered
ring; [0085] or where A.sup.1 is an alkylene group, and K is
N(R.sup.10)Y wherein R.sup.10 is alkyl, said R.sup.10 alkyl group
may be combined with said A.sup.1 alkylene group to form a
N-containing heterocyclic 5- or 6-membered ring; or a
pharmaceutically acceptable salt thereof.
[0086] It is recognized that various stereoisomeric forms of the
compounds of Formula I may exist. Preferred compounds of the
invention have any Aaa groups being .alpha.-amino acids in the
L-configuration. However, racemates and individual enantiomers and
mixtures thereof form part of the present invention.
[0087] The carbon atom designated as "M" in the compounds of
Formula I can exist in either the D or the L configuration. In some
preferred embodiments, M is a carbon atom having the "D"
configuration.
[0088] As used herein, the term "alkyl" includes straight-chain,
and branched hydrocarbon groups such as, for example, methyl,
ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl,
pentyl, 1-ethylpentyl, hexyl, and octyl groups. "Cycloalkyl" groups
are cyclic alkyl groups, such as, for example, cyclopropyl,
methylcyclopentyl, and cyclohexyl groups. Preferred alkyl groups
have 1 to about 10 carbon atoms, most preferably "lower alkyl" of 1
to about 6 carbon atoms. "Alkylene" groups are alkyl groups having
two points of attachment; i.e., non-terminal alkyl groups. "Lower
alkylene" groups are branched or unbranched alkylene groups of 1 to
about 6 carbon atoms such as, for example,
ethylene(--CH.sub.2CH.sub.2--), propylene, butylene, hexylene,
1-methylethylene, 2-methylethylene, and 2-methylpropylene.
"Cycloalkylene" groups are cyclic alkylene groups. "Acyl" groups
are alkylcarbonyl groups. "Aryl" groups are aromatic cyclic
compounds preferably including but not limited to phenyl, tolyl,
naphthyl, anthracyl, phenanthryl, and pyrenyl. Also included within
the definition of "aryl" are ring systems having two aromatic rings
connected by a bond, such as biphenyl. Preferred aryl groups
include phenyl and naphthyl.
[0089] The term "carbocyclic", as used herein, refers to cyclic
groups in which the ring portion is composed solely of carbon
atoms. The term "halogen" refers to F, Cl, Br, and I atoms. The
term "arylalkyl" denotes alkyl groups which bear aryl groups, for
example, benzyl groups. As used herein, "alkoxy" groups are alkyl
groups linked through an oxygen atom. Examples of alkoxy groups
include methoxy (--OCH.sub.3) and ethoxy (--OCH.sub.2CH.sub.3)
groups. In general, the term "oxy" when used as a suffix denotes
attachment through an oxygen atom. Thus, alkoxycarbonyl groups are
carbonyl groups which contain an alkoxy substituent, i.e., groups
of general formula --C(.dbd.O)--O--R, where R is alkyl. The term
"alkoxyalkyl" denotes an alkoxy group attached to an alkyl group.
The term "aryloxy" denotes an aryl group linked through an oxygen
atom, and the term "arylalkyloxy" denotes an arylalkyl group linked
through an oxygen atom.
[0090] The terms "heterocycle", "heterocyclyl", and "heterocyclic"
refer to cyclic groups in which a ring portion includes at least
one heteroatom such as O, N or S. Heterocyclic groups include
"heteroaryl" as well as "heteroalkyl" groups. The term "heteroaryl"
denotes aryl groups having one or more hetero atoms (e.g., O, N, or
S) contained within an aromatic ring. Also included within the
definition of "heteroaryl" are ring systems having two aromatic
rings connected by a bond, where at least one of the rings contains
a hetero atom. Preferred "heteroaryl" groups include pyridyl,
pyrimidyl, pyrrolyl, furyl, thienyl, imidazolyl, triazolyl,
tetrazolyl, quinolyl, isoquinolyl, benzoimidazolyl, thiazolyl,
bipyridyl, pyridylthiophenyl, pyrimidylthiophenyl, benzimidazolyl,
isoxazolylthiophenyl, pyrazolylthiophenyl, phthalimido, and
benzothiazolyl. The term "heterocycloalkyl" denotes a heterocycle
attached through a lower alkyl group. The term "heteroarylalkyl"
denotes a heteroaryl group attached through an alkyl group. As used
herein, the term "heteroalkyl" denotes a heterocyclic group which
contains at least one saturated carbon atom in a heterocyclic ring.
Examples of heteroalkyl groups include piperidine, dihydropyridine,
and tetrahydroisoquinyl groups. The term "arylheteroalkyl" as used
herein denotes a "heteroalkyl" group connected through an aryl
group. One preferred example of an arylheteroalkyl group is
dibenzo-.gamma.-pyranyl.
[0091] As used herein, the term "amino acid" denotes a molecule
containing both an amino group and a carboxyl group. As used herein
the term "L-amino acid" denotes an .alpha.-amino acid having the
L-configuration around the .alpha.-carbon, that is, a carboxylic
acid of general formula CH(COOH)(NH.sub.2)-(side chain), having the
L-configuration. The term "D-amino acid" similarly denotes a
carboxylic acid of general formula CH(COOH)(NH.sub.2)-- (side
chain), having the D-configuration around the .alpha.-carbon. Side
chains of L-amino acids include naturally occurring and
non-naturally occurring moieties. Nonnaturally occurring (i.e.,
unnatural) amino acid side chains are moieties that are used in
place of naturally occurring amino acid sidechains in, for example,
amino acid analogs. See, for example, Lehninger, Biochemistry,
Second Edition, Worth Publishers, Inc, 1975, pages 73-75. One
representative amino acid side chain is the lysyl side chain,
--(CH.sub.2).sub.4--NH.sub.2. Other representative .alpha.-amino
acid side chains are shown below in Table 1. TABLE-US-00001 TABLE 1
CH.sub.3-- HS--CH.sub.2-- HO--CH.sub.2--
HO.sub.2C--CH(NH.sub.2)--CH.sub.2--S--S--CH.sub.2--
C.sub.6H.sub.5--CH.sub.2-- CH.sub.3--CH.sub.2--
HO--C.sub.6H.sub.4--CH.sub.2-- CH.sub.3--S--CH.sub.2--CH.sub.2--
##STR7##
CH.sub.3--CH.sub.2--S--CH.sub.2--CH.sub.2--HO--CH.sub.2--CH.sub-
.2--CH.sub.3--CH(OH)--HO.sub.2C--CH.sub.2--NHC(.dbd.O)--CH.sub.2--
##STR8## ##STR9## ##STR10##
HO.sub.2C--CH.sub.2--CH.sub.2--NH.sub.2C(.dbd.O)--CH.sub.2--CH.sub.2--(CH-
.sub.3).sub.2--CH--(CH.sub.3).sub.2--CH--CH.sub.2--CH.sub.3--CH.sub.2--CH.-
sub.2--H.sub.2N--CH.sub.2--CH.sub.2--CH.sub.2--H.sub.2N--C(.dbd.NH)--NH--C-
H.sub.2--CH.sub.2--CH.sub.2--H.sub.2N--C(.dbd.O)--NH--CH.sub.2--CH.sub.2---
CH.sub.2-- #
CH.sub.3--CH.sub.2--CH(CH.sub.3)--CH.sub.3--CH.sub.2--CH.sub.2--CH.sub.2-
--H.sub.2N--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2-- ##STR11##
##STR12##
[0092] Functional groups present in the compounds of Formula I may
contain blocking groups. Blocking groups are known per se as
chemical functional groups that can be selectively appended to
functionalities, such as hydroxyl groups, amino groups, thio
groups, and carboxyl groups. Protecting groups are blocking groups
which can be readily removed from functionalities. These groups are
present in a chemical compound to render such functionality inert
to chemical reaction conditions to which the compound is exposed.
Any of a variety of protecting groups may be employed with the
present invention. Examples of such protecting groups are the
benzyloxycarbonyl (Cbz; Z), toluenesulfonyl, t-butoxycarbonyl,
methyl ester, and benzyl ether groups. Other preferred protecting
groups according to the invention may be found in Greene, T. W. and
Wuts, P. G. M., "Protective Groups in Organic Synthesis" 2d. Ed.,
Wiley & Sons, 1991, which is hereby incorporated by reference
in its entirety.
[0093] Further blocking groups useful in the compounds of the
present invention include those that bear acyl, aroyl, alkyl,
alkanesulfonyl, arylalkanesulfonyl, or arylsulfonyl substituents on
their amino groups. Other useful blocking groups include alkyl
ethers, e.g., the methyl ether of serine.
[0094] The disclosed compounds of the invention are useful for the
inhibition of cysteine proteases and serine proteases. As used
herein, the terms "inhibit" and "inhibition" mean having an adverse
effect on enzymatic activity. An inhibitory amount is an amount of
a compound of the invention effective to inhibit a cysteine and/or
serine protease.
[0095] Pharmaceutically acceptable salts of the cysteine and serine
protease inhibitors also fall within the scope of the compounds as
disclosed herein. The term "pharmaceutically acceptable salts" as
used herein means an inorganic acid addition salt such as
hydrochloride, sulfate, and phosphate, or an organic acid addition
salt such as acetate, maleate, fumarate, tartrate, and citrate.
Examples of pharmaceutically acceptable metal salts are alkali
metal salts such as sodium salt and potassium salt, alkaline earth
metal salts such as magnesium salt and calcium salt, aluminum salt,
and zinc salt. Examples of pharmaceutically acceptable organic
amine addition salts are salts with morpholine and piperidine.
Examples of pharmaceutically acceptable amino acid addition salts
are salts with lysine, glycine, and phenylalanine.
[0096] Compounds provided herein can be formulated into
pharmaceutical compositions by admixture with pharmaceutically
acceptable nontoxic excipients and carriers. As noted above, such
compositions may be prepared for use in parenteral administration,
particularly in the form of liquid solutions or suspensions; or
oral administration, particularly in the form of tablets or
capsules; or intranasally, particularly in the form of powders,
nasal drops, or aerosols; or dermally, via, for example,
transdermal patches; or prepared in other suitable fashions for
these and other forms of administration as will be apparent to
those skilled in the art.
[0097] The composition may conveniently be administered in unit
dosage form and may be prepared by any of the methods well known in
the pharmaceutical art, for example, as described in Remington's
Pharmaceutical Sciences (Mack Pub. Co., Easton, Pa., 1980).
Formulations for parenteral administration may contain as common
excipients sterile water or saline, polyalkylene glycols such as
polyethylene glycol, oils and vegetable origin, hydrogenated
naphthalenes and the like. In particular, biocompatible,
biodegradable lactide polymer, lactide/glycolide copolymer, or
polyoxyethylene-polyoxypropylene copolymers may be useful
excipients to control the release of the active compounds. Other
potentially useful parenteral delivery systems for these active
compounds include ethylene-vinyl acetate copolymer particles,
osmotic pumps, implantable infusion systems, cyclodextrins and
liposomes. Formulations for inhalation administration contain as
excipients, for example, lactose, or may be aqueous solutions
containing, for example, polyoxyethylene-9-lauryl ether,
glycocholate and deoxycholate, or oily solutions for administration
in the form of nasal drops, or as a gel to be applied intranasally.
Formulations for parenteral administration may also include
glycocholate for buccal administration, a salicylate for rectal
administration, or citric acid for vaginal administration.
Formulations for transdermal patches are preferably lipophilic
emulsions.
[0098] The materials for this invention can be employed as the sole
active agent in a pharmaceutical or can be used in combination with
other active ingredients which could facilitate inhibition of
cysteine and serine proteases in diseases or disorders.
[0099] The concentrations of the compounds described herein in a
therapeutic composition will vary depending upon a number of
factors, including the dosage of the drug to be administered, the
chemical characteristics (e.g., hydrophobicity) of the compounds
employed, and the route of administration. In general terms, the
compounds of this invention may be provided in effective inhibitory
amounts in an aqueous physiological buffer solution containing
about 0.1 to 10% w/v compound for parenteral administration.
Typical dose ranges are from about 1 .mu.g/kg to about 1 g/kg of
body weight per day; a preferred dose range is from about 0.01
mg/kg to 100 mg/kg of body weight per day. Such formulations
typically provide inhibitory amounts of the compound of the
invention. The preferred dosage of drug to be administered is
likely, however, to depend on such variables as the type or extent
of progression of the disease or disorder, the overall health
status of the particular patient, the relative biological efficacy
of the compound selected, and formulation of the compound
excipient, and its route of administration.
[0100] As used herein, the term "contacting" means directly or
indirectly causing at least two moieties to come into physical
association with each other. Contacting thus includes physical acts
such as placing the moieties together in a container, or
administering moieties to a patient. Thus, for example
administering a compound of the invention to a human patient
evidencing a disease or disorder associated with abnormal and/or
aberrant activity of such proteases falls within the scope of the
definition of the term "contacting".
[0101] The invention is further illustrated by way of the following
examples which are intended to elucidate the invention. These
examples are not intended, nor are they to be construed, as
limiting the scope of the disclosure.
EXAMPLES
[0102] General Methods. Thin layer chromatography was performed on
silica gel plates (MK6F 60A, size 1.times.3 in, layer thickness 250
.mu.m, Whatman Inc.). Preparative thin layer chromatography was
performed on silica gel plates (size 20.times.20 in, layer
thickness 1000 micron, Analtech). Preparative column chromatography
was carried out using Merck silica gel, 40-63 .mu.m, 230-400 mesh.
.sup.1H NMR spectra were recorded on a GE QE Plus instrument (300
MHZ) using tetramethylsilane as internal standard. Electrospray
mass spectra were recorded on a VG platform II instrument (Fisons
Instruments).
[0103] Compounds of the invention were prepared following one of
the General Methods A, B, C or D.
Example 1
[0104] Preparation of Compound 9 by General Method A ##STR13##
Preparation of Compound 1
[0105] This compound and related hydroxy acids used in this study
were synthesized following a general procedure of Harbeson et al,
J. Med. Chem. 1994, 37, 2918-2929, which is incorporated herein by
reference in its entirety.
Preparation of Compound 2
[0106] To a cooled (-10.degree. C.) solution of compound 1 (4.30 g,
0.015 mol) in anhydrous methanol (50 mL) was added slowly thionyl
chloride (3.20 mL). After 0.5 hour, the cooling bath was removed,
the mixture was stirred for an additional 16 hours and concentrated
to give a residue which on trituration with ethyl acetate (30 mL)
gave a white solid. The solid was separated by filtration and dried
to give 3.50 g of compound 2 which was used directly in the next
step; MS m/e 210(M+H).
Preparation of Compound 3
[0107] The preparation of this compound is shown in General Method
E.
Preparation of Compound 4
[0108] To a cooled (0.degree. C.) solution of compound 1 (1.00 g,
0.0034 mol) in anhydrous DMF (20 mL) was added NMM (1.40 g, 0.014
mmol) followed by 1-HOBt (0.54 g, 0.0040 mmol) and BOP (1.80 g,
0.0040 mmol). The mixture was stirred for 15 minutes and to it was
added compound 3 (0.75 g, 0.0032 mmol). The cooling bath was
removed and the mixture was stirred for 4 hours, poured into
ice-water (200 mL), and extracted into ethyl acetate (3.times.100
mL). The organic layer was washed with 2% citric acid solution
(2.times.50 mL), 2% sodium bicarbonate solution (2.times.50 mL),
brine (1.times.50 mL), and it was dried over anhydrous sodium
sulfate. Solvent evaporation under reduced pressure gave a crude
solid which was washed several times with n-pentane to produce 1.30
g of compound 4.
[0109] Compound 4: white solid (diastereomeric mixture);
.sup.1H-NMR (DMSO-d.sub.6) .delta. 7.90 and 7.65 (2 sets of t, 1H),
7.75 (d, 2H), 7.55 (q, 2H), 7.15 (m, 6H), 6.55 and 5.80 (2 sets of
d, 1H), 3.90 (m, 2H), 3.30 (d, 1H), 3.10 (m, 2H), 2.75 (m, 2H),
2.50 (m, 3H), 1.20 (s, 9H). MS m/e 478(M+H), 500(M+Na).
Preparation of Compound 5
[0110] To a solution of compound 4 (0.40 g, 0.84 mmol) in
1,4-dioxane (15 mL) was added 4 N HCl in dioxane (15 mL). The
reaction mixture was stirred at room temperature for 2 hours, then
concentrated at reduced pressure to give a residue which was washed
several times with ethyl acetate and dried under vacuum to give
0.30 g of compound 5; .sup.1H-NMR (DMSO-d.sub.6) showed complete
absence of tBoc peak at .delta. 1.20 ppm; MS m/e 378 (M+H). This
material was used directly in the next step.
Preparation of Compound 7
[0111] To a mixture of D-Ser(Bn) (compound 6, 1.00 g, 5 mmol) and 1
N NaOH (10 mL, 10 mmol) at 0.degree. C. was slowly added
methanesulfonyl chloride (0.80 g, 7.69 mmol). After 0.5 hour, the
cooling bath was removed, the mixture was stirred overnight and
acidified (pH .about.2-3) with 2 N HCl. The aqueous layer was
extracted into ethyl acetate (3.times.50 mL). The combined organic
layer was washed with water (1.times.20 mL) and brine (1.times.20
mL), and dried over MgSO.sub.4. Solvent evaporation gave a residue
which was redissolved in methylene chloride (10 mL); addition of
hexanes produced a white solid which was filtered and dried to give
1.02 g of compound 7.
Preparation of Compound 8
[0112] This compound was prepared by coupling compound 7 and
compound 5, using NMM/HOBt/BOP as coupling agents, following the
procedure described above for the preparation of compound 4. In
some of the related examples, EDCI/HOBt were used as coupling
agents.
Preparation of Compound 9
[0113] To a cooled (0.degree. C.) solution of compound 8 (0.31 g,
0.49 mmol) in anhydrous methylene chloride (10 mL) was added
Dess-Martin periodinane reagent (0.425 g, 1.00 mmol). The cooling
bath was removed and the mixture was stirred for an additional 1
hour. The solution was then diluted with methylene chloride (10
mL), and washed with 10% sodium thiosulfate solution (5.times.5
mL), saturated sodium bicarbonate solution (2.times.5 mL), and
brine (1.times.5 mL), and dried over anhydrous sodium sulfate.
Solvent removal under reduced pressure gave a residue which was
washed with n-pentane (10 mL) and dried under vacuum to produce
0.178 g of compound 9; .sup.1H-NMR spectrum revealed a minor amount
of epimerization had taken place.
[0114] Compound 9: white solid; .sup.1H-NMR (DMSO-d.sub.6) .delta.
8.75 (t, 1H), 8.60 and 8.50 (2 doublets, 1H), 7.75 (d, 2H),
7.65-7.00 (a series of m, 15H), 5.25 (broad m, 1H), 4.45 and 4.235
(2 singlets, 2H), 4.15 (m, 1H), 3.35-2.60 (a series of m, 8H), 3.35
and 3.25 (2 singlets, 3H) MS m/e 631(M+H), 653(M+Na).
Example 2
[0115] Preparation of Compound 13 by General Method B ##STR14##
[0116] In General Method B, compound 4, prepared as described
above, was oxidized by Dess-Martin periodinane reagent to generate
compound 10 which on tBoc-deprotection (2 N HCl in dioxane)
produced the amine-salt, compound 11. Coupling (NMM/HOBt/BOP) of
compound 11 with N-phenylsulfonyl-(L)-Pro (compound 12) yielded
compound 13. Purification was achieved by passing a solution of the
crude material in methylene chloride through Sep-Pak.RTM. Vac 6 cc
(1 g) silica cartidge (Waters Corporation, Milford, Mass.), eluting
with methylene chloride, followed by various combinations of
methylene chloride and ethyl acetate. Harbeson et al. (J. Med.
Chem. 1994, 37, 2918-2929) reported that silica gel chromatography
of a ketoamide epimerizes the chiral center at P.sub.1.
[0117] Compound 13: white solid; .sup.1H-NMR (CDCl.sub.3) .delta.
7.90-7.00 (a series of m, 18H), 5.40 and 5.30 (2 multiplets, 1H),
4.10 (m, 1H), 3.50-3.00 (m, 8H), 1.90-1.40 (m, 4H). MS m/e
613(M+H), 635(M+Na).
Example 3
Preparation of Compound 17 by General Method C
[0118] In General Method C, compound 2 was coupled (NMM/HOBt/BOP)
with L-Cbz-Leu to give compound 14 which was hydrolyzed (aq. NaOH)
to compound 15. Coupling (NMM/HOBt/BOP) of compound 15 with
compound 3 gave compound 16 which underwent Dess-Martin oxidation
to generate compound 17. ##STR15##
[0119] Compound 17: white solid; .sup.1H-NMR (CDCl.sub.3) .delta.
7.85 (d, 2H), 7.60-7.00 (a series of m, 15H), 6.60 (d, 1H), 5.40
(m, 1H), 5.20 (q, 1H), 5.10 (s, 2H), 4.10 (broad, 1H), 3.50-3.00 (a
series of m, 6H), 1.65-1.30 (m, 3H), 0.90 (d, 6H). MS m/e 623(M+H),
645(M+Na).
Example 4
Preparation of Compound 22 by General Method D
[0120] In General Method D, compound 7 was coupled (NMM/HOBt/BOP)
with compound 2 to generate compound 18 which underwent Dess-Martin
oxidation to generate compound 19. Hydrolysis (LiOH,
MeOH--H.sub.2O) of compound 19 gave compound 20 which was coupled
(NMM/HOBt/BOP) with compound 21 to give compound 22. Compound 22
was purified by silica gel chromatography.
[0121] Compound 22: white solid; MS m/e 646(M+H), 668(M+Na).
##STR16## Preparation of Intermediates
Example 5
[0122] The preparation of a representative example of an amine
(compound 3), containing a terminal sulfonamide moiety, is shown in
General Method E. ##STR17## Preparation of Compound 24
[0123] To a solution of 1,2-ethylenediamine (compound 23, 10.80 g,
12.00 mL, 0.18 mol) in THF (30 mL) was added slowly BOC-ON (22.10
g, 0.09 mol) in THF (70 mL) over a period of 4 hours. The reaction
mixture was stirred overnight, concentrated on a rotavapor, and
taken up into water (150 mL). The aqueous layer was acidified (pH
.about.5-6) with solid citric acid monohydrate, washed with ether
(3.times.50 mL) and then treated (at 0.degree. C.) with 6 N NaOH
solution to make it basic (pH .about.12-13). The basic solution was
extracted into ethyl acetate (3.times.100 mL), and the combined
ethyl acetate layer was dried (MgSO.sub.4) and concentrated to
generate 7.23 g of monoprotected diamine, compound 24.
[0124] Compound 24: viscous liquid; .sup.1H-NMR (CDCl.sub.3)
.delta. 5.00 (broad, 1H), 3.20 (broad q, 2H), 2.80 (t, 2H), 1.45
(s, 9H), 1.25 (broad, 2H).
Preparation of Compound 25
[0125] A cooled (0-5.degree. C.) solution of compound 24 (0.321 g,
0.002 mol) in methylene chloride (5 mL) was treated sequentially
with triethylamine (0.243 g, 0.33 mL, 0.0024 mol) and
benzenesulfonyl chloride (0.423 g, 0.30 mL, 0.0024 mol). The
ice-bath was removed and the mixture was stirred for an additional
0.5 hour, washed successively with water (2.times.5 mL), cold
(0-5.degree. C.) 0.5 N HCl (1.times.5 mL), 2% NaHCO.sub.3 solution
(1.times.5 mL), and brine (1.times.5 mL). The solution was dried
(MgSO.sub.4) and the solvent was evaporated to give a residue which
was washed several times with n-pentane. A total of 0.60 g of the
sulfonamide derivative, compound 25, was obtained.
[0126] Compound 25: white solid, mp 92-95.degree. C.; R.sub.f (TLC,
5% methanol in methylene chloride) 0.55; .sup.1H-NMR (CDCl.sub.3)
.delta. 7.85 (d, 2H), 7.55 (m, 3H), 5.30 (broad d, 1H), 4.85
(broad, 1H), 3.25 (broad q, 2H), 3.10 (broad q, 2H), 1.40 (s,
9H).
Preparation of Compound 3
[0127] A solution of compound 25 (0.560 g, 0.0019 mol) in
1,4-dioxane (4 mL) was treated with 4 N HCl in dioxane (4 mL). The
mixture was stirred at room temperature for 1 hour and concentrated
at the rotavapor. The residue was washed several times with ethyl
acetate and dried under vacuum to give 0.40 g of compound 3.
[0128] Compound 3: white solid, mp 178-180.degree. C.; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 8.20-8.00 (broad t, 4H), 7.80 (d, 2H), 7.60
(m, 3H), 2.95 (broad q, 2H), 2.80 (broad, 2H).
Example 6
Preparation of Compound 28
[0129] The preparation of a representative example of an
intermediate amine (compound 28) containing a terminal biaryl
sulfonamide moiety is shown in General Method F. ##STR18##
[0130] A mixture of compound 26 (prepared from compound 24 and
5-bromothiophene-2-sulfonyl chloride, following the same general
procedure as described above for the preparation of compound 25,
0.50 g, 1 eqv), dimethoxyethane (10 mL), 2 M Na.sub.2CO.sub.3 (5
eqv), phenylboronic acid (1.40 eqv) and Pd(PPh.sub.3).sub.4 (0.04
eqv) was heated at 135.degree. C. for 2.5 hours. The reaction
mixture was concentrated at the rotavapor, and the residue was
taken up into water (20 mL). The aqueous layer was acidified with
citric acid and extracted into methylene chloride (3.times.20 mL).
The combined organic layer was washed with water (1.times.10 mL)
and brine (1.times.10 mL). It was dried (MgSO.sub.4) and
concentrated to a small volume. Trituration of the residue with
hexanes gave a solid which was separated by filtration and dried
under vacuum to produce 0.37 g of compound 27; .sup.1H-NMR
(CDCl.sub.3) .delta. 7.60-7.20 (a series of m, 7H), 5.35 (broad,
1H), 4.85 (broad,1H), 3.30 (m, 2H), 3.20 (m, 2H), 1.40 (s, 9H). For
a general description of this reaction procedure, see Miyaura et
al., Chem. Rev. 1995, 95, 2457-2483.
[0131] Compound 27 was converted to compound 28 following the
procedure described for the preparation of compound 3.
Example 7
Preparation of Taurine Sulfonamide Intermediate
[0132] The preparation of a representative taurine sulfonamide
intermediate is shown in General Method G. ##STR19##
[0133] The phthalimide of taurine, prepared by a known procedure
(R. Winterbottom et al., J. Amer. Chem. Soc., 1947, 69, 1393-1401)
was converted to its sulfonyl chloride with phosphorous
pentachloride in refluxing benzene. This was allowed to react with
aniline in the presence of pyridine to form the corresponding
sulfonamide. The phthalimide protecting group was then removed by
refluxing with hydrazine and the resulting taurine sulfonamide was
isolated as its hydrochloride.
Example 8
[0134] Syntheses of compounds 29 through 50 in Tables 2 and 3 were
carried out using the designated general methods, as described, and
the appropriate starting materials.
Example 9
Inhibition of Cysteine Protease Activity
[0135] To evaluate inhibitory activity, stock solutions (40 times
concentrated) of each compound to be tested were prepared in 100%
anhydrous DMSO and 5 .mu.l of each inhibitor preparation were
aliquoted into each of three wells of a 96-well plate. Recombinant
human calpain I, prepared by the method of Meyer et al. (Biochem.
J. 1996, 314: 511-519; incorporated herein by reference in its
entirety), was diluted into assay buffer (i.e., 50 mM Tris, 50 mM
NaCl, 1 mM EDTA, 1 mM EGTA, and 5 mM .beta.-mercaptoethanol, pH
7.5, including 0.2 mM Succ-Leu-Tyr-MNA), and 175 .mu.l was
aliquoted into the same wells containing the independent inhibitor
stocks as well as to positive control wells containing 5 .mu.l
DMSO, but no compound. To start the reaction, 20 .mu.l of 50 mM
CaCl.sub.2 in assay buffer was added to all wells of the plate,
excepting three, which were used as background signal baseline
controls. Substrate hydrolysis was monitored every 5 minutes for a
total of 30 minutes. Substrate hydrolysis in the absence of
inhibitor was linear for up to 15 minutes.
[0136] Inhibition of calpain I activity was calculated as the
percent decrease in the rate of substrate hydrolysis in the
presence of inhibitor relative to the rate in its absence.
Comparison between the inhibited and control rates was made within
the linear range for substrate hydrolysis. The IC.sub.50s of
inhibitors (concentration yielding 50% inhibition) were determined
from the percent decrease in rates of substrate hydrolysis in the
presence of five to seven different concentrations of the test
compound. The results were plotted as percent inhibition versus log
inhibitor concentration, and the IC.sub.50 was calculated by
fitting the data to the four-parameter logistic equation shown
below using the program GraphPad Prism (GraphPad Software, Inc.,
San Diego, Calif.). y=d+[(a-d)/(1+(x/c).sup.b)]
[0137] The parameters a, b, c, and d are defined as follows: a is %
inhibition in the absence of inhibitor, b is the slope, c is the
IC.sub.50, and d is the % inhibition at an infinite concentration
of inhibitor.
[0138] Results are presented in Tables 2, 3, 4 and 5 below.
TABLE-US-00002 TABLE 2 Inhibitory Activity of Linear
.alpha.-Ketoamides ##STR20## Cmp. Calpain Prep. No. W R.sub.2 R
IC.sub.50 nM Method MS (M + 1).sup.+ 9 Ms-D-Ser(Bn) Bn Ph 16 A 631
13 PhSO.sub.2-L-Pro Bn Ph (78%)** B 613 17 Cbz-L-Leu Bn Ph 11 C 623
22 Ms-D-Ser(Bn) Bn ##STR21## 42 D 646 29 Ms-D-Ser(Bn) Bn ##STR22##
14 A 714 30 PhSO.sub.2-L-Phe Bn Ph (97%)** B 663 31 Ms-D-Ser(Bn) Bn
##STR23## 29 A 632 32 Ms-D-Ser(Bn) Bn ##STR24## 10 A 704 33
Ms-D-Ser(Bn) Bn ##STR25## 17 A 761 34 Ms-D-Ser(Bn) Bn ##STR26## 25
A 786 (M + 2).sup.+ 35 Ms-D-Ser(Bn) Bn CH.sub.3 91 A 569 36*
Ms-D-Ser(Bn) CH.sub.2OMe ##STR27## 14 A 668 37 Ms-D-Ser(Bn) Bn
##STR28## 18 C 777 38* Ms-D-Ser(Bn) CH.sub.2OMe Ph (100%)* A 585 39
Ms-D-Ser(Bn) Bn ##STR29## 22 C 715, 717 .sup.79Br, .sup.81Br 40
Ms-D-Ser(Bn) Bn ##STR30## 63 D 722 41 Ms-D-Ser(Bn) Bn ##STR31##
(88%)** D 699 42 Ms-D-Ser(Bn) Bn ##STR32## 14 C 712(M).sup.+ 43
Ms-D-Ser(Bn) Bn ##STR33## 11 C 718(M).sup.+ 44 Ms-D-Ser(Bn) Bn
##STR34## 31 D 729 (M + 2).sup.+ 45 Ms-D-Ser(Bn) Bn ##STR35## 64 D
688 46 Ms-D-Ser(Bn) Bn ##STR36## 144 D 660 *2:1 ratio
diastereomers. **Percent inhibition @ 10 .mu.M
[0139] TABLE-US-00003 TABLE 3 Inhibitory Activity of Branched-Chain
.alpha.-Ketoamides ##STR37## Ex. Calpain Prep. MS No. D IC.sub.50
nM Method (M + 1) 47 ##STR38## 26 A 645 48 ##STR39## 43 A 645 49
##STR40## 32 A 646 (M + 2) 50 CH.sub.2CH.sub.2N(CH.sub.3)SO.sub.2Ph
(100%)** C 645
[0140] Compounds listed in Table 4 were prepared by the general
methods A-G described above. TABLE-US-00004 TABLE 4 Inhibitory
Activity of .alpha.-Ketoamides ##STR41## Ex. Calpain MS No. W R
IC.sub.50 nM (M + 1) 51 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2--(3-(2-NH.sub.2-thiazol-4-yl)Ph) 29 729
(Bn) 52 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2--(5-(3-formylphenyl)thiophene-2- 5 741
(Bn) yl) 53 Ms-D-Ser CH.sub.2CH.sub.2NHC(.dbd.N--CN)OPh 15 635 (Bn)
54 Ms-D-Ser CH.sub.2CH.sub.2NHSO.sub.2(5-(3- 12 770 (Bn)
(Me.sub.2NCH.sub.2)phenyl)thiophene-2-yl) 55 Ms-D-Ser
3-Boc-NH-cyclohexane 42 645 (Bn) 56 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(5-(3- 18 812 (Bn)
(morpholinoCH.sub.2)phenyl)thiophene-2-yl) 57 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(4-(MorpholinoCH.sub.2)Ph) 18 730 (Bn) 58
Ms-D-Ser CH.sub.2CH.sub.2NHSO.sub.2(5-(3-(N-Me-piperazinyl- 21 825
(Bn) CH.sub.2)phenyl)thiophene-2-yl) 59 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(5-(3-(HOCH.sub.2)phenyl)thiophene- 35
765 (Bn) 2-yl) (2 diast) (M + Na) 60 Ms-D-Ser
CH.sub.2CH.sub.2SO.sub.2NHPh 47 631 (Bn) 61 Ms-D-Ser
CH.sub.2CH.sub.2SO.sub.2NH(4-CF.sub.3Ph) 32 699 (Bn) 62 Ms-D-Ser
(CH.sub.2).sub.3SO.sub.2NHPh 18 645 (Bn) 63 Ms-D-Ser
(CH.sub.2).sub.3SO.sub.2NH(4-CF.sub.3Ph) 23 713 (Bn) 64 Ms-D-Ser
6-ketopiperidin-3-yl (33)* 545 (Bn) 65 Ms-D-Ser
CH.sub.2CH.sub.2N(Me)SO.sub.2-(5-(3- 19 755 (Bn)
formylphenyl)thiophene-2-yl) 66 Ms-D-Thr
CH.sub.2CH.sub.2NHSO.sub.2(5-pyrid-2-ylthiophene-2-yl) 12 728 (Bn)
67 Ms-D-Ser --N(Me)SO.sub.2-(5-isoxazol-3-yl-thiophene-2-yl) 21 718
(Bn) 68 Ms-(D,L)-
CH.sub.2CH.sub.2NHSO.sub.2(5-pyrid-2-ylthiophene-2-yl) 21 670
Phenylgly 69 Ms-(D,L)- CH.sub.2CH.sub.2NHSO.sub.2Ph 80 587
Phenylgly 70 Ms-D-Thr CH.sub.2CH.sub.2NHSO.sub.2(5-(3- 23 826 (Bn)
(morpholinoCH.sub.2)phenyl)thiophene-2-yl) (Mixture of
diastereomers) 71 Ms-D-Phe
CH.sub.2CH.sub.2NHSO.sub.2(5-pyrid-2-ylthiophene-2-yl) 18 684 72
Ms-D-Ser CH.sub.2CH.sub.2NHSO.sub.2(5-(3-Fluorophenyl)thiophene-2-
18 731 (Bn) yl) (Mixture of diastereomers) 73 Ms-D-Ser
(CH.sub.2).sub.3SO.sub.2NHOCH.sub.3 87 597 (Bn) (M-1) 74 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2-(5-(3-Nitrophenyl)thiophene-2- 15 758
(Bn) yl) (Mixture of diastereomers) 75 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2-(5-(3-Methylphenyl)thiophene-2- 36 727
(Bn) yl) (Mixture of diastereomers) 76 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(5-(3-(AcNH)phenyl)thiophene-2- 11 792
(Bn) yl) (M + Na) (Mixture of diastereomers) 77 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(5-(3-(CH.sub.3CO)phenyl)thiophene- 10
777 (Bn) 2-yl) (M + Na) (Mixture of diastereomers) 78 Ms-D-Ser
1-(4- 48 770 (Bn) (Morpholinomethyl)benzenesulfonyl)pipe
ridin-4-yl) (Mixture of diastereomers) 79 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2((4-(CH.sub.3COPh)piperazin- 11 847 (Bn)
1-yl)CH.sub.2Ph) (Mixture of diastereomers) 80 Ms-D-Ser
(CH.sub.2).sub.3SO.sub.2NH-morpholin-4-yl 169 652 (Bn) (M-1) 81
Ms-D-Ser (CH.sub.2).sub.3SO.sub.2-morpholin-4-yl 124 639 (Bn) 82
Ms-D-Ser CH.sub.2CH.sub.2NHSO.sub.2-(5-(4- 13 765 (Bn)
Methoxyphenyl)thiophene-2-yl) (M + Na) (Mixture of diastereomers)
83 Ms-D-Ser CH.sub.2CH.sub.2CH.sub.2-Saccharin 48 657 (Bn) 84
Ms-D-Ser CH.sub.2CH.sub.2NHSO.sub.2((4-(PhCH.sub.2)piperazin-1- 23
819 (Bn) yl)CH.sub.2Ph) 85 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2((4-(CH.sub.3CO)piperazin-1- 14 771 (Bn)
yl)CH.sub.2Ph) 86 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2-(5-Me.sub.2N-naphth-1-yl) 49 724 (Bn) 87
Ms-D-Ser CH.sub.2CH.sub.2NHSO.sub.2-benzothiophene-2-yl 23 687 (Bn)
88 Cbz-Leu-Leu CH.sub.2CH.sub.2NHSO.sub.2(5-pyrid-2-ylthiophene-2-
33 819 yl) 89 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2((4-Pyrid-2-yl)piperazin- 21 806 (Bn)
1-yl)CH.sub.2Ph) (Mixture of diastereomers) 90 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2-(5-(4- 17 741 (Bn)
formylphenyl)thiophene-2-yl) 91 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(4-(2- 19 758 (Bn)
(MeOCH.sub.2)PyrrolidinylCH.sub.2)Ph) 92 Ms-D-Ser
(CH.sub.2).sub.5NHSO.sub.2(5-pyrid-2-ylthiophene-2- 12 756 (Bn) yl)
(Mixture of diastereomers) 93 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(5-(2- 40 812 (Bn)
(morpholinoCH.sub.2)phenyl)thiophene-2-yl) 94 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(5-(4- 22 812 (Bn)
(morpholinoCH.sub.2)phenyl)thiophene-2-yl) 95 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(5-(3- 30 810 (Bn)
(piperidinylCH.sub.2)phenyl)thiophene-2-yl) 96 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(2-acetamido-4- 23 709 (Bn)
methylthiazol-5-yl) 97 Ms-D-Ser CH.sub.2CH.sub.2NHSO.sub.2(1- 32
671 (Bn) phenylsulfonylpiperidin-4-yl) 98 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2-(5-(2- 24 741 (Bn)
formylphenyl)thiophene-2-yl) 99 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2((CH.sub.3O)CH.sub.3NCH.sub.2Ph) 21 704
(Bn) (Mixture of diastereomers) 100 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(4-ethylpiperazin-1- 21 756 (Bn)
yl)CH.sub.2Ph) (Mixture of diastereomers) 101 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(5-(3- 22 798 (Bn)
(Et.sub.2NCH.sub.2)phenyl)thiophene-2-yl) 102 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(5-(3- 36 838 (Bn)
(Cyclohexyl(Me)NCH.sub.2)phenyl)thiophene -2-yl) 103 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(5-(3- 24 796 (Bn)
(pyrrolidinylCH.sub.2)phenyl)thiophene-2-yl) 104 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(5-(3- 10 738 (Bn)
cyanophenyl)thiophene-2-yl) 105 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(4-(4- 14 816 (Bn)
acetamidophenoxy)CH.sub.2Ph) (M + Na) 106 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(5-(3- 44 782 (Bn)
(azetidinylCH.sub.2)phenyl)thiophene-2-yl) 107 Ms-D-Ser
1-(5-pyridin-2-ylthiophene-2-yl- 23 754 (Bn)
SO.sub.2)Piperidin-4-yl) (Mixture of diastereomers) 108 Ms-D-Ser
CONHCH.sub.2CH.sub.2NHSO.sub.2(5-(3-(N-ethyl-N- 10 784 (Bn)
methylaminomethyl)phenyl)thiophene-2- yl) 109 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(5-(3-(bis(2- 22 858 (Bn)
methoxyethyl)aminomethyl)phenyl)thiophene-2- yl) 110 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(5-(3-cyanophenyl)thiophene-2- 11 738
(Bn) yl) (Mixture of diastereomers) 111 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(4-(3-pyrrolin-1-yl)CH.sub.2Ph) 73 712
(Bn) (Mixture of diastereomers) 112 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2((4-(CH.sub.3SO.sub.2)piperazin-1- 37 807
(Bn) yl)CH.sub.2Ph) 113 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2((4-pyrimid-2-yl)piperazin-1- 24 807 (Bn)
yl)CH.sub.2Ph 114 Ms-D-Ser CH.sub.2CH.sub.2NHSO.sub.2(5-(3- 33 828
(Bn) (thiomorpholinoCH.sub.2)phenyl)thiophene-2-yl) 115 Ms-D-Ser
CH.sub.2CH.sub.2NHSO.sub.2(5-(3-(4- 16 824 (Bn)
ketopiperidinylCH.sub.2)phenyl)thiophene-2-yl) 116 Ms-L-Ser
CH.sub.2CH.sub.2NHSO.sub.2Ph 100 631 (Bn) *Percent inhibition @ 0.1
.mu.M Ms = methylsulfonyl
[0141] TABLE-US-00005 TABLE 5 Inhibitory Activity of Achiral
P.sub.2 Mimetic .alpha.-Ketoamides ##STR42## Ex. Calpain Synthesis
MS No. Q R IC.sub.50 nM Method (M + 1).sup.+ 117 Benzoyl ##STR43##
800 A 563 118 2,6-Dichlorobenzoyl ##STR44## 36 A 631, 633 119
2,6-Dichloro-3- methylbenzoyl ##STR45## 61 A 645 120
2,6-Difluorobenzoyl ##STR46## 20 A 599 121 2,4,6-Trifluorobenzoyl
##STR47## 85 A 618 122 2,3,4,5,6- Pentafluorobenzoyl ##STR48## 28 A
653 123 3,4- Methylenedioxybenzoyl ##STR49## >1000 A 607 124
2,5-Dichlorobenzoyl ##STR50## 68 A 631, 633 125 2-Chloro-5-
methoxybenzoyl ##STR51## 65 A 627 126 3,5-
bis(trifluoromethyl)benzoyl ##STR52## 600 A 699 127
2,6-Dimethylbenzoyl ##STR53## 178 A 591 128 2,6-Dichloronicotinoyl
##STR54## 80 A 634, 636 129 2,6-Dichlorobenzoyl ##STR55## 21 A 658,
660 130 2,6-Dichlorobenzoyl ##STR56## 20 A 687, 689 131
2,6-Dichlorobenzoyl ##STR57## 29 A 729, 731 132 2,6-Dichlorobenzoyl
##STR58## 83 A 723, 725 133 2,6-Dichlorobenzoyl ##STR59## 11 A
655,657 134 2,6-Dichlorobenzoyl ##STR60## 100 A 688, 690 135
2,6-Difluorobenzoyl ##STR61## 22 A 645 (M + Na) 136
2,6-Difluorobenzoyl ##STR62## 62 A 611 (M + Na) 137
2,6-Diethylbenzoyl ##STR63## 145 A 631 (M + Na) 138
2,6-Dimethoxybenzoyl ##STR64## 4000 A 613 139 2-Isopropylbenzoyl
##STR65## 168 A 595 140 2-Chloro-6- fluorobenzoyl ##STR66## 58 A
605 141 2-Fluoro-6- trifluoromethylbenzoyl ##STR67## 58 A 639 142
2,3,4,5,6- Pentafluorobenzoyl ##STR68## 32 A 643 143
2-Methylpropanoyl ##STR69## 1500 A 529 144 3-Methylbutanoyl
##STR70## 590 A 543 145 4-Methylpentanoyl ##STR71## 29 A 557 146
3-Cyclopentylpropanoyl ##STR72## 1000 A 583 147 E-3-Hexenoyl
##STR73## 1000 A 555 148 4-Phenylpentanoyl ##STR74## 87 A 641 (M +
Na) 149 4-Phenylbutanoyl ##STR75## 1500 A 627 (M + Na) 150
4-Methylpentanoyl ##STR76## 15 A 603 (M + Na) 151
3-Cyclopentylpropanoyl ##STR77## 420 A 607
[0142] As those skilled in the art will appreciate, numerous
changes and modifications may be made to the preferred embodiments
of the invention without departing from the spirit of the
invention. It is intended that all such variations fall within the
scope of the invention.
[0143] It is intended that each of the patents, applications, and
printed publications mentioned in this specification be hereby
incorporated by reference in their entirety.
* * * * *