U.S. patent application number 10/855699 was filed with the patent office on 2004-12-02 for caspase inhibitors and uses thereof.
Invention is credited to Brenchley, Guy, Charrier, Jean-Damien, Durrant, Steven, Knegtel, Ronald, Mortimore, Michael, Studley, John R..
Application Number | 20040242494 10/855699 |
Document ID | / |
Family ID | 33490622 |
Filed Date | 2004-12-02 |
United States Patent
Application |
20040242494 |
Kind Code |
A1 |
Brenchley, Guy ; et
al. |
December 2, 2004 |
Caspase inhibitors and uses thereof
Abstract
The present invention provides a compound of formula I: 1
wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4, and R.sup.5 are as
defined herein. The present invention also provides pharmaceutical
compositions and methods using such compositions for treating a
caspase-mediated diseases and processes for preparing the compounds
of the invention.
Inventors: |
Brenchley, Guy; (Grove
Wantage, GB) ; Charrier, Jean-Damien; (Wantage,
GB) ; Durrant, Steven; (Abingdon, GB) ;
Knegtel, Ronald; (Abingdon, GB) ; Mortimore,
Michael; (Burford, GB) ; Studley, John R.;
(Abingdon, GB) |
Correspondence
Address: |
VERTEX PHARMACEUTICALS INC.
130 WAVERLY STREET
CAMBRIDGE
MA
02139-4242
US
|
Family ID: |
33490622 |
Appl. No.: |
10/855699 |
Filed: |
May 27, 2004 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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60473622 |
May 27, 2003 |
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Current U.S.
Class: |
514/340 ;
514/1.5; 514/1.7; 514/1.9; 514/13.2; 514/15.1; 514/15.4; 514/16.4;
514/16.6; 514/16.8; 514/17.8; 514/18.2; 514/19.6; 514/20.2;
514/20.8; 514/3.8; 514/346; 514/4.3; 546/268.1; 546/291 |
Current CPC
Class: |
A61P 9/10 20180101; A61P
17/14 20180101; A61P 25/16 20180101; A61P 31/14 20180101; A61P
37/06 20180101; A61P 31/00 20180101; A61P 31/18 20180101; A61P
11/00 20180101; C07D 213/75 20130101; C07D 213/82 20130101; A61P
21/02 20180101; A61P 21/04 20180101; A61P 21/00 20180101; A61P 7/00
20180101; A61P 1/04 20180101; A61P 17/04 20180101; A61P 3/10
20180101; A61P 9/00 20180101; A61P 25/08 20180101; A61P 7/02
20180101; A61P 17/00 20180101; A61P 29/00 20180101; A61P 31/06
20180101; C07D 417/12 20130101; A61P 25/32 20180101; A61P 35/00
20180101; A61P 17/10 20180101; A61P 19/06 20180101; A61P 9/04
20180101; A61P 25/28 20180101; A61P 37/04 20180101; A61P 27/02
20180101; A61P 31/12 20180101; A61P 31/04 20180101; A61P 19/08
20180101; A61P 27/06 20180101; A61P 25/00 20180101; A61P 17/06
20180101; A61P 25/04 20180101; A61P 11/16 20180101; A61P 33/02
20180101; A61P 37/00 20180101; A61P 1/18 20180101; A61P 7/04
20180101; A61P 5/14 20180101; A61P 25/02 20180101; A61P 19/00
20180101; A61P 43/00 20180101; A61P 25/14 20180101; A61P 35/02
20180101; A61P 13/12 20180101; A61P 15/00 20180101; A61P 19/02
20180101; A61P 7/06 20180101; A61P 31/20 20180101; A61P 37/08
20180101; A61P 1/16 20180101; A61P 17/02 20180101; C07D 405/12
20130101; A61P 35/04 20180101; A61P 11/06 20180101; A61P 19/10
20180101 |
Class at
Publication: |
514/019 ;
514/346; 546/268.1; 546/291 |
International
Class: |
A61K 038/04; A61K
031/4415; C07D 41/02 |
Claims
We claim:
1. A compound of formula I: 73wherein: R.sup.1 is R.sup.6C(O)--,
HC(O)--, R.sup.6SO.sub.2--, R'OC(O)--, (R.sup.6).sub.2NC(O)--,
(R.sup.6)(H)NC(O)--, R.sup.6C(O)C(O)--, R.sup.6--,
(R.sup.6).sub.2NC(O)C(O)--, (R.sup.6)(H)NC(O)C(O)--, or
R.sup.6OC(O)C(O)--; R.sup.2 is hydrogen, --CF.sub.3, -halo,
--OR.sup.7, --NO.sub.2, --OCF.sub.3, --CN, or R.sup.8; R.sup.3 is
hydrogen or (C1--C4)-aliphatic-; R.sup.4 is --COOH or --COOR.sup.8;
R.sup.5 is --CH.sub.2F or --CH.sub.2O-2,3,5,6-tetrafluorophenyl;
R.sup.6 is (C1-C12)-aliphatic-(C3-C10)-cycloaliphatic-,
(C6-C10)-aryl-, (C3-C10)-heterocyclyl-, (C5-C10)-heteroaryl-,
(C3-C10)-cycloaliphatic-(C1- -C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)-aliphatic-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-,
(C5-C10)-heteroaryl(C1-C12)-al- iphatic-, or two R.sup.6 groups
bound to the same atom form together with that atom a 3- to
10-membered aromatic or nonaromatic ring; wherein any ring is
optionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,
(C3-C10)cycloalkyl, or a (C3-C10)heterocyclyl; wherein up to 3
aliphatic carbon atoms may be replaced by a group selected from O,
N, N(R), S, SO, and SO.sub.2; and wherein R.sup.6 is substituted
with up to 6 substituents independently selected from R; R is
halogen, --OR.sup.7, --OC(O)N(R.sup.7).sub.2, --NO.sub.2, --CN,
--CF.sub.3, --OCF.sub.3, --R.sup.7, oxo, thioxo, .dbd.NR.sup.7,
.dbd.N(OR.sup.7), 1,2-methylenedioxy, 1,2-ethylenedioxy,
--N(R.sup.7).sub.2, --SR.sup.7, --SOR.sup.7, --SO.sub.2R.sup.7,
--SO.sub.2N(R.sup.7).sub.2, --SO.sub.3R.sup.7, --C(O)R.sup.7,
--C(O)C(O)R.sup.7, --C(O)C(O)OR.sup.7, --C(O)C(O)N(R.sup.7).sub.2,
--C(O)CH.sub.2C(O)R.sup.7, --C(S)R.sup.7, --C(S)OR.sup.7,
--C(O)OR.sup.7, --OC(O)R.sup.7, --C(O)N(R.sup.7).sub.2,
--OC(O)N(R.sup.7).sub.2, --C(S)N(R.sup.7).sub.2,
--(CH.sub.2).sub.0-2NHC(- O)R.sup.7,
--N(R.sup.7)N(R.sup.7)COR.sup.7, --N(R.sup.7)N(R.sup.7)C(O)OR.s-
up.7, --N(R.sup.7)N(R.sup.7)CON(R.sup.7).sub.2,
--N(R.sup.7)SO.sub.2R.sup.- 7,
--N(R.sup.7)SO.sub.2N(R.sup.7).sub.2, --N(R.sup.7)C(O)OR.sup.7,
--N(R.sup.7)C(O)R.sup.7, --N(R.sup.7)C(S)R.sup.7,
--N(R.sup.7)C(O)N(R.sup- .7).sub.2,
--N(R.sup.7)C(S)N(R.sup.7).sub.2, --N(COR.sup.7)COR.sup.7,
--N(OR.sup.7)R.sup.7, --C(.dbd.NH)N(R.sup.7).sub.2,
--C(O)N(OR.sup.7)R.sup.7, --C(.dbd.NOR.sup.7)R.sup.7,
--OP(O)(OR.sup.7).sub.2, --P(O)(R.sup.7).sub.2,
--P(O)(OR.sup.7).sub.2, or --P(O)(H)(OR.sup.7); two R.sup.7 groups
together with the atoms to which they are bound form a 3- to
10-membered aromatic or non-aromatic ring having up to 3
heteroatoms independently selected from N, N(R), O, S, SO, or
SO.sub.2, wherein the ring is optionally fused to a (C6-C10)aryl,
(C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a (C3-C10)heterocyclyl,
and wherein any ring has up to 3 substituents selected
independently from J.sub.2; or each R.sup.7 is independently
selected from: hydrogen-, (C1-C12)-aliphatic-,
(C3-C10)-cycloaliphatic-,
(C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-, (C6-C10)-aryl-,
(C6-C10)-aryl-(C1-C12)aliphatic-, (C3-C10)-heterocyclyl-,
(C6-C10)-heterocyclyl-(C1-C12)aliphatic-, (C5-C10)-heteroaryl-, or
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; wherein R.sup.7 has up to
3 substituents selected independently from J.sub.2; and J.sub.2 is
halogen, --OR.sup.7, --OC(O)N(R.sup.7).sub.2, --NO.sub.2, --CN,
--CF.sub.3, --OCF.sub.3, --R.sup.7, oxo, thioxo, .dbd.N(R.sup.7),
.dbd.NO(R.sup.7), 1,2-methylenedioxy, 1,2-ethylenedioxy,
--N(R.sup.7).sub.2, --SR.sup.7, --SOR.sup.7, --SO.sub.2R.sup.7,
--SO.sub.2N(R.sup.7).sub.2, --SO.sub.3R.sup.7, --C(O)R.sup.7,
--C(O)C(O)R.sup.7, --(O)C(O)OR.sup.7, --C(O)C(O)N(R.sup.7).sub.2,
--C(O)CH.sub.2C(O)R.sup.7, --C(S)R.sup.7, --C(S)OR.sup.7,
--C(O)OR.sup.7, --OC(O)R.sup.7, --C(O)N(R.sup.7).sub.2,
--OC(O)N(R.sup.7).sub.2, --C(S)N(R.sup.7).sub.2,
--(CH.sub.2).sub.0-2NHC(- O)R.sup.7, --N(R.sup.7)N(R.sup.7)
COR.sup.7, --N(R.sup.7)N(R.sup.7)C(O)OR.- sup.7,
--N(R.sup.7)N(R.sup.7)CON(R.sup.7).sub.2,
--N(R.sup.7)SO.sub.2R.sup- .7,
--N(R.sup.7)SO.sub.2N(R.sup.7).sub.2, --N(R.sup.7)C(O)OR.sup.7,
--N(R.sup.7)C(O)R.sup.7, --N(R.sup.7)C(S)R.sup.7,
--N(R.sup.7)C(O)N(R.sup- .7).sub.2,
--N(R.sup.7)C(S)N(R.sup.7).sub.2, --N(COR.sup.7)COR.sup.7,
--N(OR.sup.7)R.sup.7, --CN, --C(.dbd.NH)N(R.sup.7).sub.2,
--C(O)N(OR.sup.7)R.sup.7, --C(.dbd.NOR.sup.7)R.sup.7,
--OP(O)(OR.sup.7).sub.2, --P(O)(R.sup.7).sub.2,
--P(O)(OR.sup.7).sub.2, or --P(O)(H)(OR.sup.7); and R.sup.8 is
(C1-C12)-aliphatic-(C3-C10)-cycloa- liphatic-, (C6-C10)-aryl-,
(C3-C10)-heterocyclyl-, (C5-C10)-heteroaryl-,
(C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)-aliph- atic-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, or
(C5-C10)-heteroaryl(C1-C12)-aliphatic-, wherein up to 3 aliphatic
carbon atoms may be replaced with a group selected from O, N, N(R),
S, SO, and SO.sub.2; and wherein R.sup.8 is optionally substituted
with up to 6 substituents independently selected from R.
2. A compound of formula I: 74wherein: R.sup.1 is R.sup.6C(O)--,
R.sup.6SO.sub.2--, R.sup.6OC(O)--, (R.sup.6).sub.2NC(O)--,
R.sup.6C(O)C(O)--, R.sup.6--, (R.sup.6).sub.2NC(O)C(O)--, or
R.sup.6OC(O)C(O)--; R.sup.2 is hydrogen, --CF.sub.3, -halo,
--OR.sup.7, --NO.sub.2, --OCF.sub.3, --CN, or R.sup.8; R.sup.3 is
hydrogen or (C1-C4)-aliphatic-; R.sup.4 is --COOH or --COOR.sup.8;
R.sup.5 is --CH.sub.2F or --CH.sub.2O-2,3,5,6-tetrafluorophenyl;
R.sup.6 is (C1-C12)-aliphatic-(C3-C10)-cycloaliphatic-,
(C6-C10)-aryl-, (C3-C10)-heterocyclyl-, (C5-C10)-heteroaryl-,
(C3-C10)-cycloaliphatic-(C1- -C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)-aliphatic-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-,
(C5-C10)-heteroaryl(C1-C12)-al- iphatic-, or two R.sup.6 groups
bound to the same atom form together with that atom a 3- to
10-membered aromatic or nonaromatic ring; wherein any ring is
optionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,
(C3-C10)cycloalkyl, or a (C3-C10)heterocyclyl; wherein up to 3
aliphatic carbon atoms may be replaced by a group selected from O,
N, N(R), S, SO, and SO.sub.2; and wherein R.sup.6 is substituted
with up to 6 substituents independently selected from R; R is
halogen, --OR.sup.7, --OC(O)N(R.sup.7).sub.2, --NO.sub.2, --CN,
--CF.sub.3, --OCF.sub.3, --R.sup.7, oxo, thioxo,
1,2-methylenedioxy, 1,2-ethylenedioxy, --N(R.sup.7).sub.2,
--SR.sup.7, --OR.sup.7, --SO.sub.2R.sup.7,
--SO.sub.2N(R.sup.7).sub.2, --SO.sub.3R.sup.7, --C(O)R.sup.7,
--C(O)C(O)R.sup.7, --C(O)CH.sub.2C(O)R.sup.7, --C(S)R.sup.7,
--C(O)OR.sup.7, --OC(O)R.sup.7, --C(O)N(R.sup.7).sub.2,
--OC(O)N(R.sup.7).sub.2, --C(S)N(R.sup.7).sub.2,
--(CH.sub.2).sub.0-2NHC(- O)R.sup.7,
--N(R.sup.7)N(R.sup.7)COR.sup.7, --N(R.sup.7)N(R.sup.7)C(O)OR.s-
up.7, --N(R.sup.7)N(R.sup.7)CON(R.sup.7).sub.2,
--N(R.sup.7)SO.sub.2R.sup.- 7,
--N(R.sup.7)SO.sub.2N(R.sup.7).sub.2, --N(R.sup.7)C(O)OR.sup.7,
--N(R.sup.7)C(O)R.sup.7, --N(R.sup.7)C(S)R.sup.7,
--N(R.sup.7)C(O)N(R.sup- .7).sub.2,
--N(R.sup.7)C(S)N(R.sup.7).sub.2, --N(COR.sup.7)COR.sup.7,
--N(OR.sup.7)R.sup.7, --C(.dbd.NH)N(R.sup.7).sub.2,
--C(O)N(OR.sup.7)R.sup.7, --C(.dbd.NOR.sup.7)R.sup.7,
--OP(O)(OR.sup.7).sub.2, --P(O)(R.sup.7).sub.2,
--P(O)(OR.sup.7).sub.2, or --P(O)(H)(OR.sup.7); two R.sup.7 groups
together with the atoms to which they are bound form a 3- to
10-membered aromatic or non-aromatic ring having up to 3
heteroatoms independently selected from N, N(R), O, S, SO, or
SO.sub.2, wherein the ring is optionally fused to a (C6-C10)aryl,
(C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a (C3-C10)heterocyclyl,
and wherein any ring has up to 3 substituents selected
independently from J.sub.2; or each R.sup.7 is independently
selected from: hydrogen-, (C1-C12)-aliphatic-,
(C3-C10)-cycloaliphatic-,
(C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-, (C6-C10)-aryl-,
(C6-C10)-aryl-(C1-C12)aliphatic-, (C3-C10)-heterocyclyl-,
(C6-C10)-heterocyclyl-(C1-C12)aliphatic-, (C5-C10)-heteroaryl-, or
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; wherein R.sup.7 has up to
3 substituents selected independently from J.sub.2; and J.sub.2 is
halogen, --OR , --OC(O)N(R.sup.7).sub.2, --NO.sub.2, --CN,
--CF.sub.3, --OCF.sub.3, --R.sup.7, oxo, thioxo,
1,2-methylenedioxy, 1,2-ethylenedioxy, --N(R.sup.7).sub.2,
--SR.sup.7, --SOR.sup.7, --SO.sub.2R.sup.7,
--SO.sub.2N(R.sup.7).sub.2, --SO.sub.3R.sup.7, --C(O)R.sup.7,
--C(O)C(O)R , --C(O)CH.sub.2C(O)R.sup.7, --C(S)R.sup.7,
--C(O)OR.sup.7, --OC(O)R.sup.7, --C(O)N(R.sup.7).sub.2,
--OC(O)N(R.sup.7).sub.2, --C(S)N(R.sup.7).sub.2,
--(CH.sub.2).sub.0-2NHC(- O)R.sup.7,
--N(R.sup.7)N(R.sup.7)COR.sup.7, --N(R.sup.7)N(R.sup.7)C(O)OR.s-
up.7, --N(R.sup.7)N(R.sup.7)CON(R.sup.7).sub.2,
--N(R.sup.7)SO.sub.2R.sup.- 7, --N(R )SO.sub.2N(R.sup.7).sub.2,
--N(R.sup.7)C(O)OR.sup.7, --N(R.sup.7)C(O)R.sup.7,
--N(R.sup.7)C(S)R.sup.7, --N(R.sup.7)C(O)N(R.sup- .7).sub.2,
--N(R.sup.7)C(S)N(R.sup.7).sub.2, --N(COR.sup.7)COR.sup.7,
--N(OR.sup.7)R.sup.7, --CN, --C(.dbd.NH)N(R.sup.7).sub.2,
--C(O)N(OR.sup.7)R.sup.7, --C(.dbd.NOR.sup.7)R.sup.7,
--OP(O)(OR.sup.7).sub.2, --P(O)(R.sup.7).sub.2,
--P(O)(OR.sup.7).sub.2, or --P(O)(H)(OR.sup.7); and R.sup.8 is
(C1-C12)-aliphatic-(C3-C10)-cycloa- liphatic-, (C6-C10)-aryl-,
(C3-C10)-heterocyclyl-, (C5-C10)-heteroaryl-,
(C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)-aliph- atic-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, or
(C5-C10)-heteroaryl(C1-C12)-aliphatic-, wherein up to 3 aliphatic
carbon atoms may be replaced with a group selected from O, N, N(R),
S, SO, and SO.sub.2.
3. The compound according to claim 1 or claim 2, wherein R.sup.5 is
--CH.sub.2O-2,3,5,6-tetrafluorophenyl.
4. The compound according to claim 1 or claim 2, wherein R.sup.5 is
--CH.sub.2F.
5. The compound according to any one of claims 1-4, wherein R.sup.1
is R.sup.6C(O)--.
6. The compound according to any one of claims 1-4, wherein R.sup.1
is R.sup.6SO.sub.2--.
7. The compound according to any one of claims 1-4, wherein R.sup.1
is R.sup.6--.
8. The compound according to any one of claims 1-4, wherein R.sup.1
is (R.sup.6).sub.2NC(O)--.
9. The compound according to any one of claims 1-4, wherein R.sup.1
is (R.sup.6)(H)NC(O)--.
10. The compound according to any one of claims 1-4, wherein
R.sup.1 is (R.sup.6)OC(O)--.
11. The compound according to any one of claims 1-10, wherein
R.sup.6 is (C1-C4)-aliphatic-, (C3-C10)-cycloaliphatic,
(C3-C10)-heterocyclyl, (C5-C10)-heteroaryl, (C6-C10)-aryl-, or
(C6-C10)-aryl-(C1-C12)-; wherein up to 3 aliphatic carbon atoms may
be replaced by a group selected from O, N, N(R), S, SO, and
SO.sub.2; and wherein R.sup.6 is optionally substituted.
12. The compound according to any one of claims 1-10, wherein
R.sup.6 is (C1-C4)-aliphatic-, (C5-C10)-heteroaryl-, or
(C6-C10)-aryl-, wherein the heteroaryl or aryl is optionally
substituted or wherein each R.sup.6 together with the N-atom is a
(C3-C7)-cycloaliphatic group.
13. The compound according to any one of claims 1-10, wherein
R.sup.6 is (C1-C4)-aliphatic- or (C6-C10)-aryl-, wherein the aryl
is optionally substituted or wherein each R.sup.6 together with the
N-atom is a (C3-C7)-cycloaliphatic group.
14. The compound according to any one of claims 1-13, wherein the
aliphatic is (C1-C4)-alkyl-.
15. The compound according to any one of claims 1-14, wherein
R.sup.2 is hydrogen, CF.sub.3, or CH.sub.3.
16. The compound according to any one of claims 1-15, wherein
R.sup.2 is hydrogen or CF.sub.3.
17. The compound according to any one of claims 1-16, wherein
R.sup.3 is (C1-C4)-alkyl-.
18. The compound according to any one of claims 1-17, wherein
R.sup.3 is ethyl.
19. A compound selected from Table 1.
20. A pharmaceutical composition comprising: a) a compound
according to any one of claims 1-19; and b) a pharmaceutically
acceptable carrier, adjuvant or vehicle.
21. A method for treating IL-1 mediated disease, an apoptosis
mediated disease, an inflammatory disease, an autoimmune disease, a
destructive bone disorder, a proliferative disorder, an infectious
disease, a degenerative disease, a disease associated with cell
death, a viral mediated disease, or liver disease in a patient
comprising administering to the patient a therapeutically effective
amount of a compound according to any one of claims 1-19 or a
pharmaceutical composition according to claim 20.
22. A method for treating rheumatoid arthritis, osteoarthritis,
osteoporosis, systemic lupus erythematosus, scleroderma, chronic
thyroiditis, Grave's disease, myasthenia gravis, autoimmune
neutropenia, autoimmune hemolytic anemia, thrombocytopenia,
juvenile rheumatoid arthritis, gout, Behcet's syndrome, Still's
syndrome, macrophage activation syndrome, sarcoidosis, Muckle-Wells
syndrome, familial cold urticaria, chronic infantile neurological
cutaneous and articular syndrome, familial mediterranean fever,
TNFR1-Associated Periodic Syndrome (TRAPS), Hyper-IgD periodic
fever Syndrome (HIDS), Blau's syndrome, psoriasis, atopic
dermatitis, scarring, alopecia, acne vulgaris, pemphigus, asthma,
adult respiratory distress syndrome, cystic fibrosis, emphysema,
chronic bronchitis, chronic obstructive pulmonary disease,
idiopathic pulmonary fibrosis, inflammatory peritonitis,
inflammatory bowel disease, Crohn's disease, ulcerative colitis,
autoimmune gastritis, H.pylori-associated gastric and duodenal
ulcer disease, diabetes, pancreatitis, glomerulonephritis, chronic
active hepatitis, excess dietary alcohol intake disease, renal
disease, polycystic kidney disease, burns, organ apoptosis after
burn injury, haemorrhagic shock, organ failure, endometriosis,
graft vs. host disease, organ transplant rejection, leukemia,
myelodysplastic syndrome, multiple myeloma-related bone disorder,
acute myelogenous leukemia, chronic myelogenous leukemia,
metastatic melanoma, Kaposi's sarcoma, multiple myeloma, chronic
heart disease, acute heart disease, myocardial infarction,
myocardial ischemia, congestive heart failure, atherosclerosis,
coronary artery bypass graft (CABG), a complication associated with
a coronary artery bypass graft, acute coronary syndrome,
Alzheimer's disease, Parkinson's disease, Huntington's disease,
Kennedy's disease, prion disease, cerebral ischemia, epilepsy,
spinal muscular atrophy, amyotrophic lateral sclerosis, multiple
sclerosis, HIV-related encephalitis, traumatic brain injury, spinal
cord injury, neurological damage due to stroke, diabetic
neuropathy, acute and chronic pain, uveitis, retinal disorders,
diabetic retinopathy, glaucoma, keratitis, viral mediated disease,
sepsis, septic shock, shigellosis, hepatitis-B, hepatitis-C,
hepatitis-G, yellow fever, dengue fever, Japanese encephalitis, HIV
infection, tuberculosis, meningitis, Pseudomonas infection,
Acinetobacter infection, or aging in a patient comprising
administering to the patient a therapeutically effective amount of
a compound according to any one of claims 1-19 or a pharmaceutical
composition according to claim 20.
23. The method according to claim 22, wherein the disease is
osteoarthritis, pancreatitis, asthma, adult respiratory distress
syndrome, glomerulonephritis, rheumatoid arthritis, systemic lupus
erythematosus, scleroderma, chronic thyroiditis, Grave's disease,
autoimmune gastritis, insulin-dependent diabetes mellitus (Type I),
autoimmune hemolytic anemia, autoimmune neutropenia,
thrombocytopenia, chronic active hepatitis, myasthenia gravis,
inflammatory bowel disease, Crohn's disease, psoriasis, graft vs.
host disease, osteoporosis, multiple myeloma-related bone disorder,
acute myelogenous leukemia, chronic myelogenous leukemia,
metastatic melanoma, Kaposi's sarcoma, multiple myeloma, sepsis,
septic shock, Shigellosis, cerebral ischemia, myocardial ischemia,
spinal muscular atrophy, or neurological damage due to stroke.
24. The method according to claim 22, wherein said disease is a
complication associated with a coronary artery bypass graft.
25. A method for inhibiting a caspase-mediated function in a
patient comprising administering to the patient a therapeutically
effective amount of a compound according to any one of claims 1-19
or a pharmaceutical composition according to claim 20.
26. The method for decreasing IGIF or IFN-.gamma. production in a
patient comprising administering a pharmaceutically effective
amount of a compound according to any one of claims 1-19 or a
pharmaceutical composition according to claim 20.
27. A method of preserving cells, said method comprising the step
of bathing the cells in a solution of the compound according to any
one of claims 1-19.
28. The method according to claim 27, wherein said cells are in: a)
an organ intended for transplant; or b) a blood product.
29. A method of treating cancer using immunotherapy, wherein said
immunotherapy comprises as a component thereof a compound according
to any one of claims 1-19.
30. The method according to any one of claims 20-29 wherein the
method comprises administering an additional therapeutic agent.
31. A process for preparing a compound of formula (I): 75R.sup.1 is
R.sup.6C(O)--, HC(O)--, R.sup.6SO.sub.2--, R.sup.6OC(O)--,
(R.sup.6).sub.2NC(O)--, (R.sup.6)(H)NC(O)--, R.sup.6C(O)C(O)--,
R.sup.6--, (R.sup.6).sub.2NC(O)C(O)--, (R.sup.6)(H)NC(O)C(O)--, or
R.sup.6C(O)C(O)--; R.sup.2 is hydrogen, --CF.sub.3, -halo,
--OR.sup.7, --NO.sub.2, --OCF.sub.3, --CN, or R.sup.8; R.sup.3 is
hydrogen or (C1-C4)-aliphatic-; R.sup.4 is --COOH or --COOR.sup.8;
R.sup.5 is --CH.sub.2F or --CH.sub.2O-2,3,5,6-tetrafluorophenyl;
R.sup.6 is (C1-C12)-aliphatic-(C3-C10)-cycloaliphatic-,
(C6-C10)-aryl-, (C3-C10)-heterocyclyl-, (C5-C10)-heteroaryl-,
(C3-C0l)-cycloaliphatic-(C1- -C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)-aliphatic-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-,
(C5-C10)-heteroaryl(C1-C12)-al- iphatic-, or two R.sup.6 groups
bound to the same atom form together with that atom a 3- to
10-membered aromatic or nonaromatic ring; wherein any ring is
optionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,
(C3-C10)cycloalkyl, or a (C3-C10)heterocyclyl; wherein up to 3
aliphatic carbon atoms may be replaced by a group selected from O,
N, N(R), S, SO, and SO.sub.2; and wherein R.sup.6 is substituted
with up to 6 substituents independently selected from R; R is
halogen, --OR.sup.7, --OC(O)N(R.sup.7).sub.2, --NO.sub.2, --CN,
--CF.sub.3, --OCF.sub.3, --R.sup.7, oxo, thioxo, .dbd.NR.sup.7,
.dbd.N(OR.sup.7), 1,2-methylenedioxy, 1,2-ethylenedioxy,
--N(R.sup.7).sub.2, --SR.sup.7, --SOR.sup.7, --SO.sub.2R.sup.7,
--SO.sub.2N(R.sup.7).sub.2, --SO.sub.3R.sup.7, --C(O)R.sup.7,
--C(O)C(O)R.sup.7, --C(O)C(O)OR.sup.7, --C(O)C(O)N(R.sup.7).sub.2,
--C(O)CH.sub.2C(O)R.sup.7, --C(S)R.sup.7, --C(S)OR.sup.7,
--C(O)OR.sup.7, --OC(O)R.sup.7, --C(O)N(R.sup.7).sub.2,
--OC(O)N(R.sup.7).sub.2, --C(S)N(R.sup.7).sub.2,
--(CH.sub.2).sub.0-2NHC(- O)R.sup.7,
--N(R.sup.7)N(R.sup.7)COR.sup.7, --N(R.sup.7)N(R.sup.7)C(O)OR.s-
up.7, --N(R.sup.7)N(R.sup.7)CON(R.sup.7).sub.2,
--N(R.sup.7)SO.sub.2R.sup.- 7,
--N(R.sup.7)SO.sub.2N(R.sup.7).sub.2, --N(R.sup.7)C(O)OR.sup.7,
--N(R.sup.7)C(O)R.sup.7, --N(R.sup.7)C(S)R.sup.7,
--N(R.sup.7)C(O)N(R.sup- .7).sub.2,
--N(R.sup.7)C(S)N(R.sup.7).sub.2, --N(COR.sup.7)COR.sup.7,
--N(OR.sup.7)R.sup.7, --C(.dbd.NH)N(R.sup.7).sub.2,
--C(O)N(OR.sup.7)R.sup.7, --C(.dbd.NOR.sup.7)R.sup.7,
--OP(O)(OR.sup.7).sub.2, --P(O)(R.sup.7).sub.2,
--P(O)(OR.sup.7).sub.2, or --P(O)(H)(OR.sup.7); two R.sup.7 groups
together with the atoms to which they are bound form a 3- to
10-membered aromatic or non-aromatic ring having up to 3
heteroatoms independently selected from N, N(R), O, S, SO, or
SO.sub.2, wherein the ring is optionally fused to a (C6-C10)aryl,
(C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a (C3-C10)heterocyclyl,
and wherein any ring has up to 3 substituents selected
independently from J.sub.2; or each R.sup.7 is independently
selected from: hydrogen-, (C1-C12)-aliphatic-,
(C3-C10)-cycloaliphatic-,
(C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-, (C6-C10)-aryl-,
(C6-C10)-aryl-(C1-C12)aliphatic-, (C3-C10)-heterocyclyl-,
(C6-C10)-heterocyclyl-(C1-C12)aliphatic-, (C5-C10)-heteroaryl-, or
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; wherein R.sup.7 has up to
3 substituents selected independently from J.sub.2; and J.sub.2 is
halogen, --OR.sup.7, --OC(O)N(R.sup.7).sub.2, --NO.sub.2, --CN,
--CF.sub.3, --OCF.sub.3, --R.sup.7, oxo, thioxo, .dbd.NR.sup.7,
.dbd.N(OR.sup.7), 1,2-methylenedioxy, 1,2-ethylenedioxy,
--N(R.sup.7).sub.2, --SR.sup.7, --SOR.sup.7, --SO.sub.2R.sup.7,
--SO.sub.2N(R.sup.7).sub.2, --SO.sub.3R.sup.7, --C(O)R.sup.7,
--C(O)C(O)R.sup.7, --C(O)C(O)OR.sup.7, --C(O)C(O)N(R.sup.7).sub.2,
--C(O)CH.sub.2C(O)R.sup.7, --C(S)R.sup.7, --C(S)OR.sup.7,
--C(O)OR.sup.7, --OC(O)R.sup.7, --C(O)N(R.sup.7).sub.2,
--OC(O)N(R.sup.7).sub.2, --C(S)N(R.sup.7).sub.2,
--(CH.sub.2).sub.0-2NHC(- O)R.sup.7,
--N(R.sup.7)N(R.sup.7)COR.sup.7, --N(R.sup.7)N(R.sup.7)C(O)OR.s-
up.7, --N(R.sup.7)N(R.sup.7)CON(R.sup.7).sub.2,
--N(R.sup.7)SO.sub.2R.sup.- 7,
--N(R.sup.7)SO.sub.2N(R.sup.7).sub.2, --N(R.sup.7)C(O)OR.sup.7,
--N(R.sup.7)C(O)R.sup.7, --N(R.sup.7)C(S)R.sup.7,
--N(R.sup.7)C(O)N(R.sup- .7).sub.2, --N(R.sup.7)C(S)N(R ).sub.2,
--N(COR.sup.7)COR.sup.7, --N(OR.sup.7)R.sup.7, --CN,
--C(.dbd.NH)N(R.sup.7).sub.2, --C(O)N(OR.sup.7)R.sup.7,
--C(.dbd.NOR.sup.7)R.sup.7, --OP(O)(OR.sup.7).sub.2,
--P(O)(R.sup.7).sub.2, --P(O)(OR.sup.7).sub.2, or
--P(O)(H)(OR.sup.7); and R.sup.8 is
(C1-C12)-aliphatic-(C3-C10)-cycloa- liphatic-, (C6-C10)-aryl-,
(C3-C10)-heterocyclyl-, (C5-C10)-heteroaryl-,
(C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)-aliph- atic-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, or
(C5-C10)-heteroaryl(C1-C12)-aliphatic-, wherein up to 3 aliphatic
carbon atoms may be replaced with a group selected from O, N, N(R),
S, SO, and SO.sub.2; and wherein R.sup.8 is optionally substituted
with up to 6 substituents independently selected from R;
comprising: (a) reacting a compound of formula (III): 76wherein:
R.sup.9 is --NO.sub.2, --C(O)OR.sup.10, --CN, R.sup.6C(O)N(H)--,
R.sup.6SO.sub.2N(H)--, R.sup.6OC(O)N(H)--,
(R.sup.6).sub.2NC(O)N(H)--, R.sup.6C(O)C(O)N(H)--, R.sup.6N(H)--,
(R.sup.6).sub.2NC(O)C(O)N(H)--, or R.sup.6OC(O)C(O)N(H)--; R.sup.10
is independently hydrogen, (C1-C12)-aliphatic-(C3-C10)-cycloalip-
hatic-, (C6-C10)-aryl-, (C3-C10)-heterocyclyl-,
(C5-C10)-heteroaryl-, (C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)-aliph- atic-,
(C3-C10)-heterocyclyl-(C0-C12)-aliphatic-,
(C5-C10)-heteroaryl(C1-C- 12)-aliphatic-, wherein up to 3 aliphatic
carbon atoms may be replaced with a group selected from O, N(H),
N(R), S, SO, and SO.sub.2; and wherein R.sup.10 is optionally
substituted with up to 6 substituents independently selected from
R; and R, R.sup.2, R.sup.3 and R.sup.6 are as defined above; with a
compound of formula (IV): 77wherein Y is either a carbonyl group or
an OH group; and R.sup.4 and R.sup.5 are as defined above; in the
presence of coupling conditions and a solvent; provided that if Y
is an OH group, then the process further comprises (b) oxidizing
the OH group to provide the compound of formula (I); and provided
that if R.sup.9 is --NO.sub.2, --C(O)OR.sup.10, or --ON, the
process comprises the further step of converting the --NO.sub.2,
--C(O)OR.sup.10, or --CN into R.sup.6C(O)N(H)--,
R.sup.6SO.sub.2N(H)--, R.sup.6OC(O)N(H)--,
(R.sup.6).sub.2NC(O)N(H)--, R.sup.6C(O)C(O)N(H)--, R.sup.6N(H)--,
(R.sup.6).sub.2NC(O)C(O)N(H)--, or R.sup.6OC(O)C(O)N(H)--.
32. The process according to claim 31, wherein the compound of
formula (III): 78wherein R.sup.2 is hydrogen, --CF.sub.3, -halo,
--OR.sup.7, --NO.sub.2, --OCF.sub.3, --CN, or R.sup.8; R.sup.3 is
hydrogen or (C1-C4)-aliphatic-; R.sup.9 is --NO.sub.2,
--C(O)OR.sup.10, --CN, R.sup.6C(O)N(H)--, R.sup.6SO.sub.2N(H)--,
R.sup.6OC(O)N(H)--, (R.sup.6).sub.2NC(O)N(H)--,
R.sup.6C(O)C(O)N(H)--, R.sup.6N(H)--,
(R.sup.6).sub.2NC(O)C(O)N(H)--, or R.sup.6OC(O)C(O)N(H)--; R.sup.6
is (C1-C12)-aliphatic-(C3-C10)-cycloaliphatic-, (C6-C10)-aryl-,
(C3-C10)-heterocyclyl-, (C5-C10)-heteroaryl-,
(C3-C10)-cycloaliphatic-(C1- -C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)-aliphatic-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-,
(C5-C10)-heteroaryl(C1-C12)-al- iphatic-, or two R.sup.6 groups
bound to the same atom form together with that atom a 3- to
10-membered aromatic or nonaromatic ring; wherein any ring is
optionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,
(C3-C10)cycloalkyl, or a (C3-C10)heterocyclyl; wherein up to 3
aliphatic carbon atoms may be replaced by a group selected from O,
N, N(R), S, SO, and SO.sub.2; and wherein R.sup.6 is substituted
with up to 6 substituents independently selected from R; R is
halogen, --OR.sup.7, --OC(O)N(R.sup.7).sub.2, --NO2, --CN,
--CF.sub.3, --OCF.sub.3, --R.sup.7, oxo, thioxo, .dbd.NR.sup.7,
.dbd.N(OR.sup.7), 1,2-methylenedioxy, 1,2-ethylenedioxy,
--N(R.sup.7).sub.2, --SR.sup.7, --SOR.sup.7, --SO.sub.2R.sup.7,
--SO.sub.2N(R.sup.7).sub.2, --SO.sub.3R.sup.7, --C(O)R.sup.7,
--C(O)C(O)R.sup.7, --C(O)C(O)OR.sup.7, --C(O)C(O)N(R.sup.7).sub.2,
--C(O)CH.sub.2C(O)R.sup.7, --C(S)R.sup.7, --C(O)OR.sup.7,
--OC(O)R.sup.7, --C(O)N(R.sup.7).sub.2, --OC(O)N(R.sup.7).sub.2,
--C(S)N(R.sup.7).sub.2, --(CH.sub.2).sub.0-2NHC(- O)R.sup.7,
--N(R.sup.7)N(R.sup.7)COR.sup.7, --N(R.sup.7)N(R.sup.7)C(O)OR.s-
up.7, --N(R.sup.7)N(R.sup.7)CON(R.sup.7).sub.2,
--N(R.sup.7)SO.sub.2R.sup.- 7,
--N(R.sup.7)SO.sub.2N(R.sup.7).sub.2, --N(R.sup.7)C(O)OR.sup.7,
--N(R.sup.7)C(O)R.sup.7, --N(R.sup.7)C(S)R.sup.7,
--N(R.sup.7)C(O)N(R.sup- .7).sub.2,
--N(R.sup.7)C(S)N(R.sup.7).sub.2, --N(COR.sup.7)COR.sup.7,
--N(OR.sup.7)R.sup.7, --C(.dbd.NH)N(R.sup.7).sub.2,
--C(O)N(OR.sup.7)R.sup.7, --C(.dbd.NOR.sup.7)R.sup.7,
--OP(O)(OR.sup.7).sub.2, --P(O)(R.sup.7).sub.2,
--P(O)(OR.sup.7).sub.2, or --P(O)(H)(OR.sup.7); two R.sup.7 groups
together with the atoms to which they are bound form a 3- to
10-membered aromatic or non-aromatic ring having up to 3
heteroatoms independently selected from N, N(R), O, S, SO, or
SO.sub.2, wherein the ring is optionally fused to a (C6-C10)aryl,
(C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a (C3-C10)heterocyclyl,
and wherein any ring has up to 3 substituents selected
independently from J.sub.2; or each R.sup.7 is independently
selected from: hydrogen-, (C1-C12)-aliphatic-,
(C3-C10)-cycloaliphatic-,
(C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-, (C6-C10)-aryl-,
(C6-C10)-aryl-(C1-C12)aliphatic-, (C3-C10)-heterocyclyl-,
(C6-C10)-heterocyclyl-(C1-C12)aliphatic-, (C5-C10)-heteroaryl-, or
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; wherein R.sup.7 has up to
3 substituents selected independently 10 from J.sub.2; and J.sub.2
is halogen, --OR.sup.7, --OC(O)N(R.sup.7).sub.2, --NO.sub.2, --CN,
--CF.sub.3, --OCF.sub.3, 13 R.sup.7, oxo, thioxo, .dbd.NR.sup.7,
.dbd.N(OR.sup.7), 1,2-methylenedioxy, 1,2-ethylenedioxy,
--N(R.sup.7).sub.2, --SR.sup.7, --SOR.sup.7, --SO.sub.2R.sup.7,
--SO.sub.2N(R.sup.7).sub.2, --SO.sub.3R.sup.7, --C(O)R.sup.7,
--C(O)C(O)R.sup.7, --C(O)C(O)OR.sup.7, --C(O)C(O)N(R.sup.7).sub.2,
--C(O)CH.sub.2C(O)R.sup.7, --C(S)R.sup.7, --C(S)OR.sup.7,
--C(O)OR.sup.7, --OC(O)R.sup.7, --C(O)N(R.sup.7).sub.2,
--OC(O)N(R.sup.7).sub.2, --C(S)N(R.sup.7).sub.2,
--(CH.sub.2).sub.0-2NHC(O)R.sup.7, --N(R.sup.7)N(R.sup.7)COR.sup.7,
--N(R.sup.7)N(R.sup.7)C(O)OR.sup.7,
--N(R.sup.7)N(R.sup.7)CON(R.sup.7).sub.2,
--N(R.sup.7)SO.sub.2R.sup.7, --N(R.sup.7) SO.sub.2N(R.sup.7).sub.2,
--N(R.sup.7)C(O)OR.sup.7, --N(R.sup.7)C(O)R.sup.7,
--N(R.sup.7)C(S)R.sup.7, --N(R.sup.7)C(O)N(R.sup- .7).sub.2,
--N(R.sup.7)C(S)N(R.sup.7).sub.2, --N(COR.sup.7)COR.sup.7,
--N(OR.sup.7)R.sup.7, --CN, --C(.dbd.NH)N(R.sup.7).sub.2,
--C(O)N(OR.sup.7)R.sup.7, --C(.dbd.NOR.sup.7)R.sup.7,
--OP(O)(OR.sup.7).sub.2, --P(O)(R.sup.7).sub.2,
--P(O)(OR.sup.7).sub.2, or --P(O)(H)(OR.sup.7); and R.sup.8 is
(C1-C12)-aliphatic-(C3-C10)-cycloa- liphatic-, (C6-C10)-aryl-,
(C3-C10)-heterocyclyl-, (C5-C10)-heteroaryl-,
(C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)-aliph- atic-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, or
(C5-C10)-heteroaryl(C1-C12)-aliphatic-, wherein up to 3 aliphatic
carbon atoms may be replaced with a group selected from O, N, N(R),
S, SO, and SO.sub.2; and wherein R.sup.8 is optionally substituted
with up to 6 substituents independently selected from R; is
prepared by a process comprising: (c) reacting a compound of
formula (V): 79wherein: R.sup.10 is independently hydrogen,
(C1-C12)-aliphatic-(C3-C1-C10)-cycloaliphatic-- , (C6-C10)-aryl-,
(C3-C10)-heterocyclyl-, (C5-C10)-heteroaryl-,
(C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)-aliph- atic-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-,
(C5-C10)-heteroaryl(C1-C- 12)-aliphatic-, wherein up to 3 aliphatic
carbon atoms may be replaced with a group selected from O , N,
N(R), S, SO, and SO.sub.2; and R, R.sup.2, R.sup.3, and R.sup.9 are
as defined above; in a solvent in the presence of deprotecting
conditions.
33. The process according to claim 32, wherein the compound of
formula (V): 80wherein R.sup.2, R.sup.3, R.sup.9, and R.sup.10 are
as defined in claim 32; is prepared by a process comprising: (d)
reacting a compound of formula (VI): 81wherein R.sup.2 and R.sup.9
are as defined in claim 32; with a compound of formula (VII):
82wherein X is a suitable leaving group; and R.sup.3 and R.sup.10
are as defined above; in the presence of a solvent and a base.
34. A process for preparing a compound of formula (VIII):
83wherein: R.sup.2 is --CF.sub.3, --Cl, --OR.sup.7, --NO.sub.2,
--OCF.sub.3, --CN, or R.sup.8; R.sup.3 is hydrogen or
(C1-C4)-aliphatic-; R.sup.9 is --NO.sub.2, --C(O)OR.sup.10, --CN,
R.sup.6C(O)N(H)--, R.sup.6SO.sub.2N(H)--, R.sup.6OC(O)N(H)--,
(R.sup.6).sub.2NC(O)N(H)--, R.sup.6C(O)C(O)N(H)--, R.sup.6N(H)--,
(R.sup.6).sub.2NC(O)C(O)N(H)--, or R.sup.6OC(O)C(O)N(H)--; R.sup.10
is independently hydrogen,
(C1-C12)-aliphatic-(C3-C10)-cycloaliphatic-, (C6-C10)-aryl-,
(C3-C10)-heterocyclyl-, (C5-C10)-heteroaryl-,
(C3-C10)-cycloaliphatic-(C1- -C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)-aliphatic-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-,
(C5-C10)-heteroaryl(C1-C12)-al- iphatic-, wherein up to 3 aliphatic
carbon atoms may be replaced with a group selected from O, N(H),
N(R), S, SO, and SO.sub.2; and wherein R.sup.10 is optionally
substituted with up to 6 substituents independently selected from
R; R.sup.6 is (C1-C12)-aliphatic-(C3-C10)-cyc- loaliphatic-,
(C6-C10)-aryl-, (C3-C10)-heterocyclyl-, (C5-C10)-heteroaryl-,
(C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)-aliphatic-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphat- ic-,
(C5-C10)-heteroaryl(C1-C12)-aliphatic-, or two RE groups bound to
the same atom form together with that atom a 3- to 10-membered
aromatic or nonaromatic ring; wherein any ring is optionally fused
to a (C6-C10)aryl, (C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a
(C3-C10)heterocyclyl; wherein up to 3 aliphatic carbon atoms may be
replaced by a group selected from O, N, N(R), S, SO, and SO.sub.2;
and wherein R.sup.6 is substituted with up to 6 substituents
independently selected from R; R is halogen, --OR.sup.7,
--OC(O)N(R.sup.7).sub.2, --NO.sub.2, --CN, --CF.sub.3, --OCF.sub.3,
--R.sup.7, oxo, thioxo, .dbd.NR.sup.7, .dbd.N(OR.sup.7),
1,2-methylenedioxy, 1,2-ethylenedioxy, --N(R.sup.7).sub.2,
--SR.sup.7, --SOR.sup.7, --SO.sub.2R.sup.7,
--SO.sub.2N(R.sup.7).sub.2, --SO.sub.3R.sup.7, --C(O)R.sup.7,
--C(O)C(O)R.sup.7, --C(O)C(O)OR.sup.7, --C(O)C(O)N(R.sup.7).sub.2,
--C(O)CH.sub.2C(O)R.sup.7, --C(S)R.sup.7, --C(S)OR.sup.7,
--C(O)OR.sup.7, --OC(O)R.sup.7, --C(O)N(R.sup.7).sub.2,
--OC(O)N(R.sup.7).sub.2, --C(S)N(R.sup.7).sub.2,
--(CH.sub.2).sub.0-2NHC(O)R.sup.7, --N(R.sup.7)N(R.sup.7)COR.sup.7,
--N(R.sup.7)N(R.sup.7)C(O)OR.sup.7,
--N(R.sup.7)N(R.sup.7)CON(R.sup.7).sub.2,
--N(R.sup.7)SO.sub.2R.sup.7, --N(R.sup.7)SO.sub.2N(R.sup.7).sub.2,
--N(R.sup.7)C(O)OR.sup.7, --N(R.sup.7)C(O)R.sup.7.sub.1,
--N(R.sup.7)C(S)R.sup.7, --N(R.sup.7)C(O)N(R.sup.7).sub.2,
--N(R.sup.7)C(S)N(R.sup.7).sub.2, --N(COR.sup.7)COR.sup.7,
--N(OR.sup.7)R.sup.7, --C(.dbd.NH)N(R.sup.7).sub- .2,
--C(O)N(OR.sup.7)R.sup.7, --C(.dbd.NOR)R.sup.7,
--OP(O)(OR.sup.7).sub.2, --P(O)(R.sup.7).sub.2, --P(O)(OR.sup.7) or
--P(O)(H)(OR.sup.7); two R.sup.7 groups together with the atoms to
which they are bound form a 3- to 10-membered aromatic or
non-aromatic ring having up to 3 heteroatoms independently selected
from N, N(R), O, S, SO, or SO.sub.2, wherein the ring is optionally
fused to a (C6-C10)aryl, (C5-C10)heteroaryl, (C3-C10)cycloalkyl, or
a (C3-C10)heterocyclyl, and wherein any ring has up to 3
substituents selected independently from J.sub.2; or each R.sup.7
is independently selected from: hydrogen-, (C1-C12)-aliphatic-,
(C3-C10)-cycloaliphatic-, (C3-C10)-cycloaliphatic-(C-
1-C12)-aliphatic-, (C6-C10)-aryl-,
(C6-C10)-aryl-(C1-C12)aliphatic-, (C3-C10)-heterocyclyl-,
(C6-C10)-heterocyclyl-(C1-C12)aliphatic-, (C5-C10)-heteroaryl-, or
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; wherein R.sup.7 has up to
3 substituents selected independently from J.sub.2; and J.sub.2 is
halogen, --OR.sup.7, --OC(O)N(R.sup.7).sub.2, --NO.sub.2, --CN,
--CF.sub.3, --OCF.sub.3, --R.sup.7, oxo, thioxo, .dbd.NR.sup.7,
.dbd.N(OR.sup.7), 1,2-methylenedioxy, 1,2-ethylenedioxy,
--N(R.sup.7).sub.2, --SR.sup.7, --SOR.sup.7, --SO.sub.2R.sup.7,
--SO.sub.2N(R.sup.7).sub.2, --SO.sub.3R.sup.7, --C(O)R.sup.7,
--C(O)C(O)R.sup.7, --C(O)C(O)OR.sup.7, --C(O)C(O)N(R.sup.7).sub.2,
--C(O)CH.sub.2C(O)R.sup.7, --C(S)R.sup.7, --C(S)OR.sup.7,
--C(O)OR.sup.7, --OC(O)R.sup.7, --C(O)N(R.sup.7).sub.2,
--OC(O)N(R.sup.7).sub.2, --C(S)N(R.sup.7).sub.2,
--(CH.sub.2).sub.0-2NHC(O)R.sup.7, --N(R.sup.7)N(R.sup.7)COR.sup.7,
--N(R.sup.7)N(R.sup.7)C(O)OR.sup.7,
--N(R.sup.7)N(R.sup.7)CON(R.sup.7).sub.2,
--N(R.sup.7)SO.sub.2R.sup.7, --N(R.sup.7)SO.sub.2N(R.sup.7).sub.2,
--N(R.sup.7)C(O)OR.sup.7, --N(R.sup.7)C(O)R.sup.7,
--N(R.sup.7)C(S)R.sup.7, --N(R.sup.7)C(O)N(R.sup- .7).sub.2,
--N(R.sup.7)C(S)N(R.sup.7).sub.2, --N(COR.sup.7)COR.sup.7,
--N(OR.sup.7)R.sup.7, --CN, --C(.dbd.NH)N(R.sup.7).sub.2,
--C(O)N(OR.sup.7)R.sup.7, --C(.dbd.NOR.sup.7)R.sup.7,
--OP(O)(OR.sup.7).sub.2, --P(O)(R.sup.7).sub.2,
--P(O)(OR.sup.7).sub.2, or --P(O)(H)(OR.sup.7); and R.sup.8 is
(C1-C12)-aliphatic-(C3-C10)-cycloa- liphatic-, (C6-C10)-aryl-,
(C3-C10)-heterocyclyl-, (C5-C10)-heteroaryl-,
(C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)-aliph- atic-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, or
(C5-C10)-heteroaryl(C1-C12)-aliphatic-, wherein up to 3 aliphatic
carbon atoms may be replaced with a group selected from O, N, N(R),
S, SO, and SO.sub.2; and wherein R.sup.8 is optionally substituted
with up to 6 substituents independently selected from R; comprising
the step of (e) reacting a compound of formula (IX): 84wherein
R.sup.2 and R.sup.9 are as defined above; with a compound of
formula (VII): 85wherein R.sup.3 and R.sup.10 are as defined above;
and X is a suitable leaving group; in the presence of-a solvent and
a base.
35. A process for preparing a compound of formula (I): 86wherein:
R.sup.1 is R.sup.6C(O)--, R.sup.6SO.sub.2--, R.sup.6OC(O)--,
(R.sup.6).sub.2NC(O)--, (R.sup.6)(H)NC(O)--, R.sup.6C(O)C(O)--,
R.sup.6--, (R.sup.6).sub.2NC(O)C(O)--, (R.sup.6)(H)NC(O)C(O)--, or
R.sup.6OC(O)C(O)--; R.sup.2 is hydrogen, --CF.sub.3, -halo,
--OR.sup.7, --NO.sub.2, --OCF.sub.3, --CN, or R.sup.8; R.sup.3 is
hydrogen or (C1-C4)-aliphatic-; R.sup.4 is --COOH or --COOR.sup.8;
R.sup.5 is --CH.sub.2F or --CH.sub.2O-2,3,5,6-tetrafluorophenyl;
R.sup.6 is (C1-C12)-aliphatic-(C3-C10)-cycloaliphatic-,
(C6-C10)-aryl-, (C3-C10)-heterocyclyl-, (C5-C10)-heteroaryl-,
(C3-C10)-cycloaliphatic-(C1- -C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)-aliphatic-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-,
(C5-C10)-heteroaryl(C1-C12)-al- iphatic-, or two R.sup.6 groups
bound to the same atom form together with that atom a 3- to
10-membered aromatic or nonaromatic ring; wherein any ring is
optionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,
(C3-C10)cycloalkyl, or a (C3-C10)heterocyclyl; wherein up to 3
aliphatic carbon atoms may be replaced by a group selected from O,
N, N(R), S, SO, and SO.sub.2; and wherein R.sup.6 is substituted
with up to 6 substituents independently selected from R; R is
halogen, --OR.sup.7, --OC(O)N(R.sup.7).sub.2, --NO.sub.2, --CN,
--CF.sub.3, --OCF.sub.3, --R.sup.7, oxo, thioxo, .dbd.NR.sup.7,
.dbd.N(OR.sup.7), 1,2-methylenedioxy, 1,2-ethylenedioxy,
--N(R.sup.7).sub.2, --SR.sup.7, --SOR.sup.7, --SO.sub.2R.sup.7,
--SO.sub.2N(R.sup.7).sub.2, --SO.sub.3R.sup.7, --C(O)R.sup.7,
--C(O)C(O)R.sup.7, --C(O)C(O)OR.sup.7, --C(O)C(O)N(R.sup.7).sub.2,
--C(O)CH.sub.2C(O)R.sup.7, --C(S)R.sup.7, --C(S)OR.sup.7,
--C(O)OR.sup.7, --OC(O)R.sup.7, --C(O)N(R.sup.7).sub.2,
--OC(O)N(R.sup.7).sub.2, --C(S)N(R.sup.7).sub.2,
--(CH.sub.2).sub.0-2NHC(- O)R.sup.7,
--N(R.sup.7)N(R.sup.7)COR.sup.7, --N(R.sup.7)N(R.sup.7)C(O)OR.s-
up.7, --N(R.sup.7)N(R.sup.7)CON(R.sup.7).sub.2,
--N(R.sup.7)SO.sub.2R.sup.- 7,
--N(R.sup.7)SO.sub.2N(R.sup.7).sub.2, --N(R.sup.7)C(O)OR.sup.7,
--N(R.sup.7)C(O)R.sup.7, --N(R.sup.7)C(S)R.sup.7,
--N(R.sup.7)C(O)N(R.sup- .7).sub.2,
--N(R.sup.7)C(S)N(R.sup.7).sub.2, --N(COR.sup.7)COR.sup.7,
--N(OR.sup.7)R.sup.7, --C(.dbd.NH)N(R.sup.7).sub.2,
--C(O)N(OR.sup.7)R.sup.7, --C(.dbd.NOR.sup.7)R.sup.7,
--OP(O)(OR.sup.7).sub.2, --P(O)(R.sup.7).sub.2,
--P(O)(OR.sup.7).sub.2, or --P(O)(H)(OR.sup.7); two R.sup.7 groups
together with the atoms to which they are bound form a 3- to
10-membered aromatic or non-aromatic ring having up to 3
heteroatoms independently selected from N, N(R), O, S, SO, or
SO.sub.2, wherein the ring is optionally fused to a (C6-C10)aryl,
(C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a (C3-C10)heterocyclyl,
and wherein any ring has up to 3 substituents selected
independently from J.sub.2; or each R.sup.7 is independently
selected from: hydrogen-, (C1-C12)-aliphatic-,
(C3-C10)-cycloaliphatic-,
(C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-, (C6-C10)-aryl-,
(C6-C10)-aryl-(C1-C12)aliphatic-, (C3-C10)-heterocyclyl-,
(C6-C10)-heterocyclyl-(C1-C12)aliphatic-, (C5-C10)-heteroaryl-, or
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; wherein R.sup.7 has up to
3 substituents selected independently from J.sub.2; and J.sub.2 is
halogen, --OR.sup.7, --OC(O)N(R.sup.7).sub.2, --NO.sub.2, --CN,
--CF.sub.3, --OCF.sub.3, --R.sup.7, oxo, thioxo, .dbd.NR.sup.7,
.dbd.N(OR.sup.7), 1,2-methylenedioxy, 1,2-ethylenedioxy,
--N(R.sup.7).sub.2, --SR.sup.7, --SOR.sup.7, --SO.sub.2R.sup.7,
--SO.sub.2N(R.sup.7).sub.2, --SO.sub.3R.sup.7, --C(O)R.sup.7,
--C(O)C(O)R.sup.7, --C(O)C(O)OR.sup.7, --C(O)C(O)N(R.sup.7).sub.2,
--C(O)CH.sub.2C(O)R.sup.7, --C(S)R.sup.7, --C(S)OR.sup.7,
--C(O)OR.sup.7, --OC(O)R.sup.7, --C(O)N(R.sup.7).sub.2,
--OC(O)N(R.sup.7).sub.2, --C(S)N(R.sup.7).sub.2,
--(CH.sub.2).sub.0-2NHC(- O)R.sup.7,
--N(R.sup.7)N(R.sup.7)COR.sup.7, --N(R.sup.7)N(R.sup.7)C(O)OR.s-
up.7, --N(R.sup.7)N(R.sup.7)CON(R.sup.7).sub.2,
-N(R.sup.7)SO.sub.2R.sup.7- , --N(R.sup.7)SO.sub.2N(R.sup.7).sub.2,
--N(R.sup.7)C(O)OR.sup.7, --N(R.sup.7)C(O)R.sup.7,
--N(R.sup.7)C(S)R.sup.7, --N(R.sup.7)C(O)N(R.sup- .7).sub.2,
--N(R.sup.7)C(S)N(R.sup.7).sub.2, --N(COR.sup.7)COR.sup.7,
--N(OR.sup.7)R.sup.7, --CN, --C(.dbd.NH)N(R.sup.7).sub.2,
--C(O)N(OR.sup.7)R.sup.7, --C(.dbd.NOR.sup.7)R.sup.7,
--OP(O)(OR.sup.7).sub.2, --P(O)(R.sup.7).sub.2,
--P(O)(OR.sup.7).sub.2, or --P(O)(H)(OR.sup.7); and R.sup.8 is
(C1-C12)-aliphatic-(C3-C10)-cycloa- liphatic-, (C6-C10)-aryl-,
(C3-C10)-heterocyclyl-, (C5-C10)-heteroaryl-,
(C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)-aliph- atic-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, or
(C5-C10)-heteroaryl(C1-C12)-aliphatic-, wherein up to 3 aliphatic
carbon atoms may be replaced with a group selected from O, N, N(R),
S, SO, and SO.sub.2; and wherein R.sup.8 is optionally substituted
with up to 6 substituents independently selected from R;
comprising: (a) reacting a compound of formula (III): 87wherein:
R.sup.9 is --NO.sub.2, --C(O)OR.sup.10, --CN, R.sup.6C(O)N(H)--,
R.sup.6SO.sub.2N(H)--, R.sup.6OC(O)N(H)--,
(R.sup.6).sub.2NC(O)N(H)--, R.sup.6C(O)C(O)N(H)--, R.sup.6N(H)--,
(R.sup.6).sub.2NC(O)C(O)N(H)--, or R.sup.6OC(O)C(O)N(H)--; and
R.sup.2, R.sup.10 and R.sup.6 are as defined above; with a compound
of formula (X): 88wherein Y is either a carbonyl group or an OH
group; and R.sup.3, R.sup.4 and R.sup.5 are as defined above; in
the presence of coupling conditions and a solvent; provided that if
Y is an OH group, then the process further comprises (b) oxidizing
the OH group to provide the compound of formula (I); and provided
that if R.sup.9 is --NO.sub.2, --C(O)OR.sup.10, or --CN, the
process comprises the further step of converting the --NO.sub.2,
--C(O)OR.sup.10, or --CN into R.sup.6C(O)N(H)--,
R.sup.6SO.sub.2N(H)--, R.sup.6OC(O)N(H)--,
(R.sup.6).sub.2NC(O)N(H)--, R.sup.6C(O)C(O)N(H)--, R.sup.6N(H)--,
(R.sup.6).sub.2NC(O)C(O)N(H)--, or R.sup.6OC(O)C(O)N(H)--.
Description
[0001] This application claims the benefit of U.S. Provisional
Application No. 60/473,622, filed May 27, 2003.
FIELD OF THE INVENTION
[0002] This invention is in the field of medicinal chemistry and
relates to compounds, and pharmaceutical compositions thereof, that
inhibit caspases that mediate cell apoptosis and inflammation. The
invention also relates to processes for preparing these compounds.
The invention further relates to methods of using the compounds and
pharmaceutical compositions of this invention to treat diseases
where caspase activity is implicated.
BACKGROUND OF THE INVENTION
[0003] Apoptosis, or programmed cell death, is a principal
mechanism by which organisms eliminate unwanted cells. The
deregulation of apoptosis, either excessive apoptosis or the
failure to undergo it, has been implicated in a number of diseases
such as cancer, acute inflammatory and autoimmune disorders,
ischemic diseases and certain neurodegenerative disorders (see
generally Science, 1998, 281, 1283-1312; Ellis et al., Ann. Rev.
Cell. Biol., 1991, 7, 663).
[0004] Caspases are a family of cysteine protease enzymes that are
key mediators in the signaling pathways for apoptosis and cell
disassembly (Thornberry, Chem. Biol., 1998, 5, R97-R103). These
signaling pathways vary depending on cell type and stimulus, but
all apoptosis pathways appear to converge at a common effector
pathway leading to proteolysis of key proteins. Caspases are
involved in both the effector phase of the signaling pathway and
further upstream at its initiation. The upstream caspases involved
in initiation events become activated and in turn activate other
caspases that are involved in the later phases of apoptosis.
[0005] Caspase-1, the first identified caspase, is also known as
interleukin converting enzyme or "ICE." Caspase-1 converts
precursor interleukin-1.beta. ("pIL-1.beta.") to the
pro-inflammatory active form by specific cleavage of pIL-1.beta.
between Asp-116 and Ala-117. Besides caspase-1 there are also
eleven other known human caspases, all of which cleave specifically
at aspartyl residues. They are also observed to have stringent
requirements for at least four amino acid residues on the
N-terminal side of the cleavage site.
[0006] The caspases have been classified into three groups
depending on the amino acid sequence that is preferred or primarily
recognized. The group of caspases, which includes caspases 1, 4, 5
and 13, have been shown to prefer hydrophobic aromatic amino acids
at position 4 on the N-terminal side of the cleavage site. Another
group which includes caspases 2, 3 and 7, recognize aspartyl
residues at both positions 1 and 4 on the N-terminal side of the
cleavage site, and preferably a sequence of Asp-Glu-X-Asp. A third
group, which includes caspases 6, 8, 9 and 10, tolerate many amino
acids in the primary recognition sequence, but seem to prefer
residues with branched, aliphatic side chains such as valine and
leucine at position 4.
[0007] The caspases have also been grouped according to their
perceived function. The first subfamily consists of caspases-1
(ICE), 4, 5 and 13. These caspases have been shown to be involved
in pro-inflammatory cytokine processing and therefore play an
important role in inflammation. Caspase-1, the most studied enzyme
of this class, activates the IL-1.beta. precursor by proteolytic
cleavage. This enzyme therefore plays a key role in the
inflammatory response. Caspase-1 is also involved in the processing
of interferon-.gamma. inducing factor (IGIF, also known as IL-18)
which stimulates the production of interferon gamma, a key
immunoregulator that modulates antigen presentation, T-cell
activation and cell adhesion.
[0008] The remaining caspases make up the second and third
subfamilies. These enzymes are of central importance in the
intracellular signaling pathways leading to apoptosis. One
subfamily consists of the enzymes involved in initiating events in
the apoptotic pathway, including transduction of signals from the
plasma membrane. Members of this subfamily include caspases-2, 8, 9
and 10. The other subfamily, consisting of the effector capsases 3,
6 and 7, are involved in the final downstream cleavage events that
result in the systematic breakdown and death of the cell by
apoptosis. Caspases involved in the upstream signal transduction
activate the downstream caspases, which then disable DNA repair
mechanisms, fragment DNA, dismantle the cell cytoskeleton and
finally fragment the cell.
[0009] Knowledge of the four amino acid sequence primarily
recognized by the caspases has been used to design caspase
inhibitors. Reversible tetrapeptide inhibitors have been prepared
having the structure
CH.sub.3CO--[P4]-[P3]-[P2]-CH(R)CH.sub.2CO.sub.2H where P2 to P4
represent an optimal amino acid recognition sequence and R is an
aldehyde, nitrile or ketone capable of binding to the caspase
cysteine sulfhydryl. Rano and Thornberry, Chem. Biol. 4, 149-155
(1997); Mjalli, et al., Bioorg. Med. Chem. Lett. 3, 2689-2692
(1993); Nicholson et al., Nature 376, 37-43 (1995). Irreversible
inhibitors based on the analogous tetrapeptide recognition sequence
have been prepared where R is an acyloxymethylketone
--COCH.sub.2OCOR'. R' is exemplified by an optionally substituted
phenyl such as 2,6-dichlorobenzoyloxy and where R is COCH.sub.2X
where X is a leaving group such as F or Cl. Thornberry et al.,
Biochemistry 33, 3934 (1994); Dolle et al., J Med. Chem. 37,
563-564 (1994).
[0010] The utility of caspase inhibitors to treat a variety of
mammalian disease states associated with an increase in cellular
apoptosis has been demonstrated using peptidic caspase inhibitors.
For example, in rodent models caspase inhibitors have been shown to
reduce infarct size and inhibit cardiomyocyte apoptosis after
myocardial infarction, to reduce lesion volume and neurological
deficit resulting from stroke, to reduce post-traumatic apoptosis
and neurological deficit in traumatic brain injury, to be effective
in treating fulminant liver destruction, and to improved survival
after endotoxic shock. Yaoita et al., Circulation, 97, 276 (1998);
Endres et al., J Cerebral Blood Flow and Metabolism, 18, 238,
(1998); Cheng et al., J. Clin. Invest., 101, 1992 (1998); Yakovlev
et al., J Neuroscience, 17, 7415 (1997); Rodriquez et al., J. Exp.
Med., 184, 2067 (1996); Grobmyer et al., Mol. Med., 5, 585
(1999).
[0011] In general, the peptidic inhibitors described above are very
potent against some of the caspase enzymes. However, this potency
has not always been reflected in cellular models of apoptosis. In
addition peptide inhibitors are typically characterized by
undesirable pharmacological properties such as poor oral
absorption, poor stability and rapid metabolism. Plattner and
Norbeck, in Drug Discovery Technologies, Clark and Moos, Eds.
(Ellis Horwood, Chichester, England, 1990).
[0012] Recognizing the need to improve the pharmacological
properties of the peptidic caspase inhibitors, peptidomimetic
inhibitors have been reported. Amongst these, inhibitors where the
P3 amino acid has been replaced by derivatives of
3-aminopyridin-2-ones and 5-aminopyrimidin-4-ones have been
reported (U.S. Pat. No. 5,756,466 (Bemis et al.); PCT Publication
No. WO 95/35308 (Bemis et al.); Dolle et al. J. Med. Chem. 39,
2438, (1996); Golec et al. Bioorg. Med. Chem. Lett. 7, 2181,
(1997); Semple et al, Biorg. Med. Chem. Lett. 7, 1337, (1997)).
[0013] Due to the inherent problems of the peptidic inhibitors,
there continues to be a need for small molecule, nonpeptide caspase
inhibitors that are potent, stable, and penetrate membranes to
provide effective inhibition of apoptosis in vivo. Such compounds
would be extremely useful in treating the aforementioned diseases
where caspase enzymes play a role.
SUMMARY OF THE INVENTION
[0014] The present invention provides a compound of formula I:
2
[0015] wherein: R.sup.1, R.sup.2, R.sup.3, R.sup.4, and R.sup.5 are
as defined herein.
[0016] The present invention also provides pharmaceutical
compositions comprising a compound of formula I and methods using
such compounds and compositions for treating caspase-mediated
diseases. The present invention also provides processes for
preparing the compounds of formula I.
DETAILED DESCRIPTION OF THE INVENTION
[0017] The present invention provides a compound of formula I:
3
[0018] wherein:
[0019] R.sup.1 is R.sup.6C(O)--, HC(O)--, R.sup.6SO.sub.2--,
R.sup.6OC(O)--, (R.sup.6).sub.2NC(O)--, (R.sup.6) (H)NC(O)--,
R.sup.6C(O)C(O)--, R.sup.6--, (R.sup.6).sub.2NC(O)C(O)--, (R.sup.6)
(H)NC(O)C(O)--, or R.sup.6OC(O)C(O)--;
[0020] R.sup.2 is hydrogen, --CF.sub.3, halo, --OR.sup.7,
--NO.sub.2, --OCF.sub.3, --CN, or R.sup.8;
[0021] R.sup.3 is hydrogen or (C1-C4)-aliphatic-;
[0022] R.sup.4 is --COOH or --COOR.sup.8;
[0023] R.sup.5 is --CH.sub.2F or
--CH.sub.2O-2,3,5,6-tetrafluorophenyl;
[0024] R.sup.6 is (C1-C12)-aliphatic-(C3-C10)-cycloaliphatic-,
(C6-C10)-aryl-, (C3-C10)-heterocyclyl-, (C5-C10)-heteroaryl-,
(C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)-aliph- atic-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-,
(C5-C10)-heteroaryl(C1-C- 12)-aliphatic-, or two R.sup.6 groups
bound to the same atom form together with that atom a 3- to
10-membered aromatic or nonaromatic ring; wherein the ring is
optionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,
(C3-C10)cycloalkyl, or a (C3-C10)heterocyclyl; wherein up to 3
aliphatic carbon atoms may be replaced by a group selected from O,
N, N (R), S,SO, and SO.sub.2; and wherein R.sup.6 is substituted
with up to 6 substituents independently selected from R;
[0025] R is halogen, --OR.sup.7, --OC(O)N(R.sup.7).sub.2,
--NO.sub.2, --CN, --CF.sub.3, --OCF.sub.3, --R.sup.7, oxo, thioxo,
.dbd.NR.sup.7, .dbd.N(OR.sup.7), 1,2-methylenedioxy,
1,2-ethylenedioxy, --N(R.sup.7).sub.2, --SR.sup.7, --SOR.sup.7,
--SO.sub.2R.sup.7, --SO.sub.2N(R.sup.7).sub.2, --SO.sub.3R.sup.7,
--C(O)R.sup.7, --C(O)C(O)R.sup.7, --C(O)C(O)OR.sup.7,
--C(O)C(O)N(R.sup.7).sub.2, --C(O)CH.sub.2C(O)R.sup.7,
--C(S)R.sup.7, --C(S)OR.sup.7, --C(O)OR.sup.7, --OC(O)R.sup.7,
--C(O)N(R.sup.7).sub.2, --OC(O)N(R.sup.7).sub.2,
--C(S)N(R.sup.7).sub.2, --(CH.sub.2).sub.0-2NHC(O)R.sup.7,
--N(R.sup.7)N(R.sup.7)COR.sup.7,
--N(R.sup.7)N(R.sup.7)C(O)OR.sup.7,
--N(R.sup.7)N(R.sup.7)CON(R.sup.7).sub.2, --N(R.sup.7)
SO.sub.2R.sup.7, --N(R.sup.7) SO.sub.2N(R.sup.7).sub.2,
--N(R.sup.7)C(O)OR.sup.7, --N(R.sup.7)C(O)R.sup.7,
--N(R.sup.7)C(S)R.sup.7, --N(R.sup.7)C(O)N(R.sup- .7).sub.2,
--N(R.sup.7)C(S)N(R.sup.7).sub.2, --N(COR.sup.7)COR.sup.7,
--N(OR.sup.7)R.sup.7, --C(.dbd.NH)N(R.sup.7).sub.2,
--C(O)N(OR.sup.7)R.sup.7, --C(.dbd.NOR.sup.7)R.sup.7,
--OP(O)(OR.sup.7).sub.2, --P(O)(R.sup.7).sub.2,
--P(O)(OR.sup.7).sub.2, or --P(O)(H)(OR.sup.7);
[0026] two R.sup.7 groups together with the atoms to which they are
bound form a 3- to 10-membered aromatic or non-aromatic ring having
up to 3 heteroatoms independently selected from N, N(R), O, S,SO,
or SO.sub.2, wherein the ring is optionally fused to a
(C6-C10)aryl, (C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a
(C3-C10)heterocyclyl, and wherein any ring has up to 3 substituents
selected independently from J.sub.2; or
[0027] each R.sup.7 is independently selected from:
[0028] hydrogen-,
[0029] (C1-C12)-aliphatic-,
[0030] (C3-C10)-cycloaliphatic-,
[0031] (C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-,
[0032] (C6-C10)-aryl-,
[0033] (C6-C10)-aryl-(C1-C12)aliphatic-,
[0034] (C3-C10)-heterocyclyl-,
[0035] (C6-C10)-heterocyclyl-(C1-C12)aliphatic-,
[0036] (C5-C10)-heteroaryl-, or
[0037] (C5-C10)-heteroaryl-(C1-C12)-aliphatic-;
[0038] wherein R.sup.7 has up to 3 substituents selected
independently from J.sub.2; and
[0039] J.sub.2 is halogen, --OR.sup.7, --OC(O)N(R.sup.7).sub.2,
--NO.sub.2, --CN, --CF.sub.3, --OCF.sub.3, --R.sup.7, oxo, thioxo,
.dbd.NR.sup.7, .dbd.NOR.sup.7, 1,2-methylenedioxy,
1,2-ethylenedioxy, --N(R.sup.7).sub.2, --SR.sup.7, --SOR.sup.7,
--SO.sub.2R.sup.7, --SO.sub.2N(R.sup.7).sub.2, --SO.sub.3R.sup.7,
--C(O)R.sup.7, --C(O)C(O)R.sup.7, --C(O)C(O)OR.sup.7,
--C(O)C(O)N(R.sup.7).sub.2, --C(O)CH.sub.2C(O)R.sup.7,
--C(S)R.sup.7, --C(S)OR.sup.7, --C(O)OR.sup.7, --OC(O)R.sup.7,
--C(O)N(R.sup.7).sub.2, --OC(O)N(R.sup.7).sub.2,
--C(S)N(R.sup.7).sub.2, --(CH.sub.2).sub.0-2NHC(O)R.sup.7,
--N(R.sup.7)N(R.sup.7)COR.sup.7,
--N(R.sup.7)N(R.sup.7)C(O)OR.sup.7,
--N(R.sup.7)N(R.sup.7)CON(R.sup.7).sub.2,
--N(R.sup.7)SO.sub.2R.sup.7, --N(R.sup.7)SO.sub.2N(R.sup.7).sub.2,
--N(R.sup.7)C(O)OR.sup.7, --N(R.sup.7)C(O)R.sup.7,
--N(R.sup.7)C(S)R.sup.7, --N(R.sup.7)C(O)N(R.sup- .7).sub.2,
--N(R.sup.7)C(S)N(R.sup.7).sub.2, --N(COR.sup.7)COR.sup.7,
--N(OR.sup.7)R.sup.7, --CN, --C(.dbd.NH)N(R.sup.7).sub.2,
--C(O)N(OR.sup.7)R.sup.7, --C(.dbd.NOR.sup.7)R.sup.7,
--OP(O)(OR.sup.7).sub.2, --P(O)(R.sup.7).sub.2,
--P(O)(OR.sup.7).sub.2, or --P(O)(H)(OR.sup.7); and
[0040] R.sup.8 is (C1-C12)-aliphatic-(C3-C10)-cycloaliphatic-,
(C6-C10)-aryl-, (C3-C10)-heterocyclyl-, (C5-C10)-heteroaryl-,
(C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)-aliph- atic-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, or
(C5-C10)-heteroaryl(C1-C12)-aliphatic-, wherein up to 3 aliphatic
carbon atoms may be replaced with a group selected from O, N, N(R),
S, SO, and SO.sub.2; and wherein
[0041] R.sup.8 is optionally substituted with up to 6 substituents
independently selected from R.
[0042] The present invention also provides a compound of formula I:
4
[0043] wherein:
[0044] R.sup.1 is R.sup.6C(O)--, R.sup.6SO.sub.2--, R.sup.6OC(O)--,
(R.sup.6).sub.2NC(O)--, R.sup.6C(O)C(O)--, R.sup.6--,
(R.sup.6).sub.2NC(O)C(O)--, or R.sup.6OC(O)C(O)--;
[0045] R.sup.2 is hydrogen, --CF.sub.3, halo, --OR.sup.7,
--NO.sub.2, --OCF.sub.3, --CN, or R.sup.8;
[0046] R.sup.3 is hydrogen or (C1-C4)-aliphatic-;
[0047] R.sup.4 is --COOH or --COOR.sup.8;
[0048] R.sup.5 is --CH.sub.2F or
--CH.sub.2O-2,3,5,6-tetrafluorophenyl;
[0049] R.sup.6 is (C1-C12)-aliphatic-(C3-C10)-cycloaliphatic-,
(C6-C10)-aryl-, (C3-C10)-heterocyclyl-, (C5-C10)-heteroaryl-,
(C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)-aliph- atic-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-,
(C5-C10)-heteroaryl(C1-C- 12)-aliphatic-, or two R.sup.6 groups
bound to the same atom form together with that atom a 3- to
10-membered aromatic or nonaromatic ring; wherein the ring is
optionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,
(C3-C10)cycloalkyl, or a (C3-C10)heterocyclyl; wherein up to 3
aliphatic carbon atoms may be replaced by a group selected from O,
N(H), N(R), S, SO, and SO.sub.2; and wherein R.sup.6 is substituted
with up to 6 substituents independently selected from R;
[0050] R is halogen, --OR.sup.7, --OC(O)N(R.sup.7).sub.2,
--NO.sub.2, --CN, --CF.sub.3, --OCF.sub.3, --R.sup.7, oxo, thioxo,
1,2-methylenedioxy, 1,2-ethylenedioxy, --N(R.sup.7).sub.2,
--SR.sup.7, --SOR.sup.7.sub.1, --SO.sub.2R.sup.7,
--SO.sub.2N(R.sup.7).sub.2, --SO.sub.3R.sup.7, --C(O)R.sup.7,
--C(O)C(O)R.sup.7, --C(O)CH.sub.2C(O)R.sup.7, --C(S)R.sup.7,
--C(O)OR.sup.7, --OC(O)R.sup.7, --C(O)N(R.sup.7).sub.2,
--OC(O)N(R.sup.7).sub.2, --C(S)N(R.sup.7).sub.2,
--(CH.sub.2).sub.0-2NHC(O)R.sup.7, --N(R.sup.7)N(R.sup.7)COR.sup.7,
--N(R.sup.7)N(R.sup.7)C(O)OR.sup.7,
--N(R.sup.7)N(R.sup.7)CON(R.sup.7).su- b.2,
--N(R.sup.7)SO.sub.2R.sup.7, --N(R.sup.7)SO.sub.2N(R.sup.7).sub.2,
--N(R.sup.7)C(O)OR.sup.7, --N(R.sup.7)C(O)R.sup.7,
--N(R.sup.7)C(S)R.sup.7, --N(R.sup.7)C(O)N(R.sup.7).sub.2,
--N(R.sup.7)C(S)N(R.sup.7).sub.2, --N(COR.sup.7)COR.sup.7,
--N(OR.sup.7)R.sup.7, --C(.dbd.NH)N(R.sup.7).sub.2,
--C(O)N(OR.sup.7)R.sup.7, --C(.dbd.NOR.sup.7)R.sup.7,
--OP(O)(OR.sup.7).sub.2, --P(O)(R ).sub.2, --P(O)(OR.sup.7).sub.2,
or --P(O)(H)(OR.sup.7);
[0051] two R.sup.7 groups together with the atoms to which they are
bound form a 3- to 10-membered aromatic or non-aromatic ring having
up to 3 heteroatoms independently selected from N(H), N(R), O, S,
SO, or SO.sub.2, wherein the ring is optionally fused to a
(C6-C10)aryl, (C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a
(C3-C10)heterocyclyl, and wherein any ring has up to 3 substituents
selected independently from J.sub.2; or
[0052] each R.sup.7 is independently selected from:
[0053] hydrogen-,
[0054] (C1-C12)-aliphatic-,
[0055] (C3-C10)-cycloaliphatic-,
[0056] (C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-,
[0057] (C6-C10)-aryl-,
[0058] (C6-C10)-aryl-(C1-C12)aliphatic-,
[0059] (C3-C10)-heterocyclyl-,
[0060] (C6-C10)-heterocyclyl-(C1-C12)aliphatic-,
[0061] (C5-C10)-heteroaryl-, or
[0062] (C5-C10)-heteroaryl-(C1-C12)-aliphatic-;
[0063] wherein R.sup.7 has up to 3 substituents selected
independently from J.sub.2; and
[0064] J.sub.2 is halogen, --OR.sup.7, --OC(O)N(R.sup.7).sub.2,
--NO.sub.2, --CN, --CF.sub.3, --OCF.sub.3, --R.sup.7, oxo, thioxo,
1,2-methylenedioxy, 1,2-ethylenedioxy, --N(R.sup.7).sub.2,
--SR.sup.7--SOR.sup.7, --SO.sub.2R.sup.7,
--SO.sub.2N(R.sup.7).sub.2, --SO.sub.3R.sup.7, --C(O)R.sup.7,
--C(O)C(O)R.sup.7, --C(O)CH.sub.2C(O)R.sup.7, --C(S)R.sup.7,
--C(O)OR.sup.7, --OC(O)R.sup.7, --C(O)N(R.sup.7).sub.2,
--OC(O)N(R.sup.7).sub.2, --C(S)N(R.sup.7).sub.2,
--(CH.sub.2).sub.0-2NHC(O)R.sup.7, --N(R.sup.7)N(R.sup.7)COR.sup.7,
--N(R.sup.7)N(R.sup.7)C(O)OR.sup.7,
--N(R.sup.7)N(R.sup.7)CON(R.sup.7).su- b.2,
--N(R.sup.7)SO.sub.2R.sup.7, --N(R.sup.7) SO.sub.2N(R.sup.7).sub.2,
--N(R.sup.7)C(O)OR.sup.7, --N(R.sup.7)C(O)R.sup.7,
--N(R.sup.7)C(S)R.sup.7, --N(R.sup.7)C(O)N(R.sup.7).sub.2,
--N(R.sup.7)C(S)N(R.sup.7).sub.2, --N(COR.sup.7)COR.sup.7,
--N(OR.sup.7)R.sup.7, --CN, --C(.dbd.NH)N(R.sup.7).sub.2,
--C(O)N(OR.sup.7)R.sup.7, --C(.dbd.NOR.sup.7)R.sup.7,
--OP(O)(OR.sup.7).sub.2, --P(O)(R.sup.7).sub.2,
--P(O)(OR.sup.7).sub.2, or --P(O)(H)(OR.sup.7); and
[0065] R.sup.8 is (C1-C12)-aliphatic-(C3-C10)-cycloaliphatic-,
(C6-C10)-aryl-, (C3-C10)-heterocyclyl-, (C5-C10)-heteroaryl-,
(C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)-aliph- atic-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, or
(C5-C10)-heteroaryl(C1-C12)-aliphatic-, wherein up to 3 aliphatic
carbon atoms may be replaced with a group selected from O, N(H),
N(R), S, SO, and SO.sub.2.
[0066] Another embodiment of this invention provides a compound
wherein, R.sup.1 is R.sup.6C(O)--, R.sup.6SO.sub.2--, or R.sup.6--.
In a preferred embodiment, R.sup.1 is R.sup.6C(O)--. In another
preferred embodiment, R.sup.1 is R.sup.6SO.sub.2--. In yet another
preferred embodiment, R.sup.1 is R.sup.6--.
[0067] Another embodiment of this invention provides a compound
wherein R.sup.1 is (R.sup.6).sub.2NC(O)--or (R.sup.6)OC(O)--. In a
preferred embodiment, R.sup.1 is (R.sup.6).sub.2NC(O)--. In another
preferred embodiment, R.sup.1 is (R.sup.6)(H)NC(O)--. In yet
another preferred embodiment, R.sup.1 is (R.sup.6)OC(O)--.
[0068] In one embodiment of this invention, each R.sup.6 is
independently (C1-C4)-aliphatic-, (C3-C10)-cycloaliphatic,
(C3-C10)-heterocyclyl, (C5-C10)-heteroaryl, (C6-C10)-aryl-, or
(C6-C10)-aryl-(C1-C12)-(it being understood that optionally up to 3
aliphatic carbon atoms may be replaced by a group selected from O,
N, N(R), S, SO, and SO.sub.2; and wherein R.sup.6 is optionally
substituted with up to 6 substituents independently selected from R
or R.sup.6 is substituted as disclosed in any of the embodiments
herein).
[0069] In another embodiment, each R.sup.6 is independently H,
(C1-C4)-aliphatic- or (C6-C10)-aryl- or each R.sup.6 together with
the N-atom is a (C3-C7)-cycloaliphatic.
[0070] In another embodiment, each R.sup.6 is independently
(C1-C4)-aliphatic-, (C5-C10)-heteroaryl-, or (C6-C10)-aryl-,
wherein the heteroaryl or aryl is optionally substituted or wherein
each R.sup.6 together with the N-atom is a (C3-C7)-cycloaliphatic
group.
[0071] In another embodiment, each R.sup.6 is independently
(C1-C4)-aliphatic- or (C6-C10)-aryl-, wherein the aryl is
optionally substituted or wherein each R.sup.6 together with the
N-atom is a (C3-C7)-cycloaliphatic.
[0072] In yet another embodiment, each R.sup.6 is independently
(C1-C4)-aliphatic-, (C3-C7)-cycloaliphatic, (C6-C10)-aryl-,
(C5-C10)-heteroaryl, wherein the heteroaryl and aryl are
independently and optionally substituted, or each R.sup.6 together
with the N-atom is a (C3-C7)-cycloaliphatic.
[0073] According to a preferred embodiment of this invention,
R.sup.2 is hydrogen, C1-, C2-, C3-, or C4-alkyl-, --CF.sub.3, --C1,
--OR.sup.7, --NO.sub.2, --OCF.sub.3, or --CN. More preferably,
R.sup.2 is hydrogen, C1-alkyl-, C2-alkyl-, or CF.sub.3. More
preferably, R.sup.2 is hydrogen or CF.sub.3.
[0074] According to another preferred embodiment, R.sup.3 is
ethyl.
[0075] According to another preferred embodiment, R.sup.5 is
--CH.sub.2O-2,3,5,6-tetrafluorophenyl.
[0076] According to another preferred embodiment, R.sup.5 is
--CH.sub.2F.
[0077] According to another preferred embodiment, R.sup.8 is
(C1-C12)-alkyl. More preferably, R.sup.8 is (C1-C4)-alkyl.
[0078] According to a preferred embodiment, each R and J.sub.2 are
independently halogen, --OR.sup.7, --OC(O)N(R.sup.7).sub.2,
--NO.sub.2, --CN, --CF.sub.3, --OCF.sub.3, --R.sup.7, oxo,
1,2-methylenedioxy, 1,2-ethylenedioxy, --N(R.sup.7).sub.2,
--C(O)R.sup.7, --C(O)C(O)R.sup.7, --C(O)OR.sup.7, --OC(O)R.sup.7,
--C(O)N(R.sup.7).sub.2, or --OC(O)N(R.sup.7).sub.2.
[0079] As used herein, the carbon atom designations may have the
indicated integer and any intervening integer. For example, the
number of carbon atoms in a (C1-C4)-alkyl group is 1, 2, 3, or 4.
It should be understood that these designation refer to the total
number of atoms in the appropriate group. For example, in a
(C3-C10)-heterocyclyl the total number of carbon atoms and
heteroatoms is 3 (as in aziridine), 4, 5, 6 (as in morpholine), 7,
8, 9, or 10.
[0080] As used herein, an aliphatic group includes straight-chained
and branched groups having the specified number of atoms. If the
number of atoms is unspecified, the aliphatic group has from 1 to
12 carbon atoms. As would be understood, alkenyl and/or alkynyl
aliphatic groups have a minimum of 2 carbon atoms. Preferred
aliphatic groups are alkyl groups (preferably having from 1 to 6
atoms).
[0081] Accordingly, unless otherwise specified, preferred aliphatic
groups of this invention are alkyl groups and have 1, 2, 3, 4, 5,
or 6 carbon atoms. More preferred alkyl groups have 1, 2, 3, or 4
carbon atoms. Preferred alkenyl and alkynyl groups of this
invention have 2, 3, 4, 5, or, 6 carbon atoms and more preferably,
from 2, 3, or 4 carbon atoms.
[0082] Cycloalkyl and cycloalkenyl groups have between 3 and 10
carbon atoms and are monocyclic or bicyclic, including linearly
fused, bridged, or spirocyclic. A cycloaliphatic group is,
preferably, a cycloalkyl or a cylcoalkenyl. More preferred
cycloaliphatic groups are 3-, 4-, 5-, 6-, or 7-membered rings that
are, more preferably, cycloalkyl rings.
[0083] As used herein, "aromatic group" or "aryl" refers to a
6-10-membered ring system that contains at least one aromatic ring.
Example of aromatic rings include phenyl and naphthyl.
[0084] As used herein a "heteroaryl" refers to ring system having
5-10 members and 1, 2, or 3 heteroatoms independently selected from
N, N(R), O, S, SO, and SO.sub.2., wherein at least one ring is
heteroaromatic (e.g., pyridyl, thiophene, or thiazole). Preferred
heteroaryl groups are 5- or 6-membered rings having 1 or 2
heteroatoms. In certain embodiments of this invention, more
preferred heteroaryl groups are those that have contain a ".dbd.N"
group.
[0085] Examples of heteroaryl rings include 2-furanyl, 3-furanyl,
N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl,
benzimidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl,
2-oxazolyl, 4-oxazolyl, 5-oxazolyl, N-pyrrolyl, 2-pyrrolyl,
3-pyrrolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl,
4-pyrimidinyl, 5-pyrimidinyl, pyridazinyl (e.g., 3-pyridazinyl),
2-thiazolyl, 4-thiazolyl, 5-thiazolyl, tetrazolyl (e.g.,
5-tetrazolyl), triazolyl (e.g., 2-triazolyl and 5-triazolyl),
2-thienyl, 3-thienyl, benzofuryl, benzothiophenyl, indolyl (e.g.,
2-indolyl), pyrazolyl (e.g., 2-pyrazolyl), isothiazolyl,
1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl,
1,2,3-triazolyl, 1,2,3-thiadiazolyl, 1,3,4-thiadiazolyl,
1,2,5-thiadiazolyl, purinyl, pyrazinyl, 1,3,5-triazinyl, quinolinyl
(e.g., 2-quinolinyl, 3-quinolinyl, 4-quinolinyl), and isoquinolinyl
(e.g., 1-isoquinolinyl, 3-isoquinolinyl, or 4-isoquinolinyl).
[0086] As used herein a."heterocycle" refers to ring system having
3-10 members and 1, 2, or 3 heteroatoms independently selected from
N, N(R), O, S, SO, and SO.sub.2, wherein no ring is aromatic (e.g.,
piperidine and morpholine). Preferred heterocyclyl groups are 5- or
6-membered rings having 1 or 2 heteroatoms.
[0087] Examples of heterocyclic rings include
3-1H-benzimidazol-2-one, 3-(1-alkyl)-benzimidazol-2-one,
2-tetrahydrofuranyl, 3-tetrahydrofuranyl, 2-tetrahydrothiophenyl,
3-tetrahydrothiophenyl, 2-morpholino, 3-morpholino, 4-morpholino,
2-thiomorpholino, 3-thiomorpholino, 4-thiomorpholino,
1-pyrrolidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl,
1-tetrahydropiperazinyl, 2-tetrahydropiperazinyl,
3-tetrahydropiperazinyl- , 1-piperidinyl, 2-piperidinyl,
3-piperidinyl, 1-pyrazolinyl, 3-pyrazolinyl, 4-pyrazolinyl,
5-pyrazolinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl,
4-piperidinyl, 2-thiazolidinyl, 3-thiazolidinyl, 4-thiazolidinyl,
1-imidazolidinyl, 2-imidazolidinyl, 4-imidazolidinyl,
5-imidazolidinyl, indolinyl, tetrahydroquinolinyl,
tetrahydroisoquinolinyl, benzothiolane, benzodithiane, and
1,3-dihydro-imidazol-2-one.
[0088] Any of these cycloaliphatic, heterocyclyl, and heteroaryl
groups are optionally fused with a 5- or 6-membered aryl or
heteroaryl ring. Furthermore, each of any aliphatic, aryl,
cycloaliphatic, heteroaryl, and heterocyclyl may contain
appropriate substituents (preferably up to 5, more preferable up to
3, and even more preferably, 0 or 1) independently selected from,
for example, carbonyl and R. Preferred substituents (including R
and J.sub.2) are halogen, --OR.sup.7, --NO.sub.2, --CF.sub.3,
--OCF.sub.3, --R.sup.7, oxo, --OR.sup.7, --O-benzyl, --O-phenyl,
1,2-methylenedioxy, 1,2-ethylenedioxy, --N(R.sup.7).sub.2,
--C(O)R.sup.7, --COOR.sup.7 or --CON(R.sup.7).sub.2, wherein
R.sup.7 is defined herein (and is preferably H, (C1-C6)-alkyl, or
(C2-C6)-alkenyl and alkynyl), with (C1-C6)-alkyl being most
preferred). It should be understood that this definition would
include a perfluorinated alkyl group.
[0089] In embodiments of this invention where R is a substituent on
a nitrogen atom, preferred R groups are selected from the group
consisting of --R.sup.7, --SOR.sup.7, --SO.sub.2R.sup.7,
--SO.sub.2N(R.sup.7).sub.2, --SO.sub.3R.sup.7, --C(O)R.sup.7,
--C(O)C(O)R.sup.7, --C(O)C(O)OR.sup.7, --C(O)C(O)N(R.sup.7).sub.2,
--C(O)CH.sub.2C(O)R.sup.7, --C(S)R.sup.7, --C(S)OR.sup.7,
--C(O)OR.sup.7, --C(O)N(R.sup.7).sub.2, --C(S)N(R.sup.7).sub.2,
--(CH.sub.2).sub.0-2NHC(O)R.sup.7, --N(R.sup.7)N(R.sup.7)COR.sup.7,
--N(R.sup.7)N(R.sup.7)C(O)OR.sup.7,
--N(R.sup.7)N(R.sup.7)CON(R.sup.7).sub.2,
--N(R.sup.7)SO.sub.2R.sup.7, --N(R.sup.7)SO.sub.2N(R.sup.7).sub.2,
--N(R.sup.7)C(O)OR.sup.7, --N(R.sup.7)C(O)R.sup.7,
--N(R.sup.7)C(S)R.sup.7, --N(R.sup.7)C(O)N(R.sup- .7).sub.2,
--N(R.sup.7)C(S)N(R.sup.7).sub.2, --N(COR.sup.7)COR.sup.7,
--N(OR.sup.7)R.sup.7, --C(.dbd.NH)N(R.sup.7).sub.2,
--C(O)N(OR.sup.7)R.sup.7, --C(.dbd.NOR.sup.7)R.sup.7,
--OP(O)(OR.sup.7).sub.2, --P(O)(R.sup.7).sub.2,
--P(O)(OR.sup.7).sub.2, and --P(O)(H)(OR.sup.7) wherein R.sup.7 is
defined herein (and is preferably H, (C1-C6)-alkyl, or
(C2-C6)-alkenyl and alkynyl), with (C1-C6)-alkyl being most
preferred). More preferably, such R groups are selected from the
group consisting of --R.sup.7 and --C(O) R.sup.7.
[0090] In preferred compounds of this invention, the
stereochemistry is as depicted below: 5
[0091] Any of the embodiments disclosed herein may be combined to
provide alternative embodiments of this invention. Specific
embodiments of this invention may be selected from the substituents
depicted in the compounds of Table 1
[0092] The compounds of the present invention are broad caspase
inhibitors and have an improved ability over reported compounds to
inhibit apoptosis (see Examples 42 and 43).
[0093] According to a preferred embodiment, this invention provides
a compound of formula Ia or Ib 6
[0094] wherein R.sup.1, R.sup.2, R.sup.3, and R.sup.4 are as
defined in any of the embodiments herein.
[0095] According to a more preferred embodiment, the compound of
formula I of present invention provides a compound of formula II,
selected from Table 1 below: 7
1TABLE 1 Compounds of the invention. Ex. R.sup.1 R.sup.2 R.sup.3
R.sup.5 1 Me(C.dbd.O)-- H Et CH.sub.2O-- 2,3,5,6-tetrafluorophenyl
2 Et(C.dbd.O)-- H Et CH.sub.2O-- 2,3,5,6-tetrafluorophenyl 3
n-Pr(C.dbd.O)-- H Et CH.sub.2O-- 2,3,5,6-tetrafluorophenyl 4
c-Pr(C.dbd.O)-- H Et CH.sub.2O-- 2,3,5,6-tetrafluorophenyl 5
i-Pr(C.dbd.O)-- H Et CH.sub.2O-- 2,3,5,6-tetrafluorophenyl 6
MeOCH.sub.2(C.dbd.O)-- H Et CH.sub.2O-- 2,3,5,6-tetrafluorophenyl 7
2-Furyl(C.dbd.O)-- H Et CH.sub.2O-- 2,3,5,6-tetrafluorophenyl 8
3-Furyl(C.dbd.O)-- H Et CH.sub.2O-- 2,3,5,6-tetrafluorophenyl 9
3-Pyridyl(C.dbd.O)-- H Et CH.sub.2O-- 2,3,5,6-tetrafluorophenyl 10
3- H Et CH.sub.2O-- 2,3,5,6-tetrafluorophenyl Isothiazole(C.dbd.O)
11 Ph(C.dbd.O)-- H Et CH.sub.2O-- 2,3,5,6-tetrafluorophenyl 12
Bn(C.dbd.O)-- H Et CH.sub.2O-- 2,3,5,6-tetrafluorophenyl 13
Me(C.dbd.O)-- CF.sub.3 Et CH.sub.2O-- 2,3,5,6-tetrafluorophenyl 14
EtNH(C.dbd.O)-- H Et CH.sub.2O-- 2,3,5,6-tetrafluorophenyl 15
(Et).sub.2N(C.dbd.O)-- H Et CH.sub.2O-- 2,3,5,6-tetrafluorophenyl
16 Pyrrolidinyl H Et CH.sub.2O-- 2,3,5,6-tetrafluorophenyl
(C.dbd.O)-- 17 MeO(C.dbd.O)-- H Et CH.sub.2O--
2,3,5,6-tetrafluorophenyl 18 Et(SO.sub.2)-- H Et CH.sub.2O--
2,3,5,6-tetrafluorophenyl 19 n-Pr(SO.sub.2)-- H Et CH.sub.2O--
2,3,5,6-tetrafluorophenyl 20 i-Pr(SO.sub.2)-- H Et CH.sub.2O--
2,3,5,6-tetrafluorophenyl 21 Ph(SO.sub.2)-- H Et CH.sub.2O--
2,3,5,6-tetrafluorophenyl 22 Et(SO.sub.2)-- CF.sub.3 Et CH.sub.2O--
2,3,5,6-tetrafluorophenyl 23 Bn(C.dbd.O) H i- CH.sub.2O--
2,3,5,6-tetrafluorophenyl Pr 24 Et(SO.sub.2) H i- CH.sub.2O--
2,3,5,6-tetrafluorophenyl Pr 25 Et(C.dbd.O) H Me CH.sub.2F 26
Ph(C.dbd.O) H Me CH.sub.2F 27 2,6-DiClPh(C.dbd.O) H Me CH.sub.2F 28
Bn(C.dbd.O) H Me CH.sub.2F 29 Et(C.dbd.O) H Et CH.sub.2F 30
Ph(C.dbd.O) H Et CH.sub.2F 31 2,6-DiClPh(C.dbd.O) H Et CH.sub.2F 32
2-Pyridyl(C.dbd.O) H Et CH.sub.2F 33 Bn(C.dbd.O) H Et CH.sub.2F 34
3-MeBn(C.dbd.O) H Et CH.sub.2F 35 Et(C.dbd.O) H n- CH.sub.2F Pr 36
Et(C.dbd.O) H i- CH.sub.2F Bu 37 Bn(C.dbd.O) Me Et CH.sub.2F 38
Thiazol-2-yl H Et CH.sub.2O-- 2,3,5,6-tetrafluorophenyl 39 n-Propyl
H Et CH.sub.2O-- 2,3,5,6-tetrafluorophenyl
[0096] According to another embodiment, the present invention
provides a pharmaceutical composition comprising:
[0097] a) a compound of formula I, as defined herein, or a
pharmaceutically acceptable salt thereof; and
[0098] b) a pharmaceutically acceptable carrier, adjuvant or
vehicle.
[0099] It will be apparent to one skilled in the art that certain
compounds of this invention may exist in tautomeric forms or
hydrated forms, all such forms of the compounds being within the
scope of the invention. Unless otherwise stated, structures
depicted herein are also meant to include all stereochemical forms
of the structure; i.e., the R and S configurations for each
asymmetric center. Therefore, single stereochemical isomers as well
as enantiomeric and diastereomeric mixtures of the present
compounds are within the scope of the invention. Unless otherwise
stated, structures depicted herein are also meant to include
compounds that differ only in the presence of one or more
isotopically enriched atoms. For example, compounds having the
present structures except for the replacement of a hydrogen by a
deuterium or tritium, or the replacement of a carbon by a
.sup.13C-- or .sup.14C-enriched carbon are within the scope of this
invention.
[0100] The compounds of this invention may be prepared in general
by methods known to those skilled in the art for analogous
compounds and by the preparative examples that follow. For the
purposes of illustration, the following Schemes I-III for the
synthesis of the compounds of the present invention are provided.
It should be understood that any protective group depicted in the
schemes may be varied as appropriate in view of compatibility with
other substituents.
[0101] Various protecting groups may be used in the methods of this
invention (see, e.g., T. W. Greene & P. G. M Wutz, "Protective
Groups in Organic Synthesis", 3.sup.rd Edition, John Wiley &
Sons, Inc. (1999) and the earlier editions of this book). Typical
functional groups that must be protected are amines. Any amines and
other functional groups may be protected according to methods known
in the art. Compounds, including amines, may be used with or
without isolation from the reaction mixtures.
[0102] Scheme I 8
[0103] In Scheme I above, the following abbreviations are used: EDC
is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide; HOBt is
1-hydroxybenzotriazole; THF is tetrahydrofuran; TFA is
trifluoroacetic acid; DCM is dichloromethane; DMAP is
4-dimethylaminopyridine. Acid 1 is coupled to amino alcohol 2. Here
the coupling is depicted using EDC/DMAP/HOBt/THF, however, other
suitable conditions may also be used. Depending on the nature of
R.sup.4 and R.sup.5 an amino ketone may be used, in place of the
amino alcohol, thus avoiding the subsequent oxidation step. In the
case of fluoromethyl ketones where R.sup.5 is CH.sub.2F, the amino
alcohol 2 may be obtained according to the method of Revesz et al.,
Tetrahedron Lett. 1994, 35, 9693. In the case of tetrafluorophenoxy
ketones where R.sup.5 is --CH.sub.2O-2,3,5,6-tetrafluo- rophenyl,
amino alcohol 2 may be obtained by methods analogous to those of
Semple et al., Bioorganic and Medicinal Chemistry Letters, 1997, 7,
1337 (Scheme II).
[0104] Finally the hydroxy group in compound 3 is oxidized (e.g.,
with Dess-Martin periodinane) and the resulting compound treated
appropriately according to the nature of R.sup.4. For example, in
product I if R.sup.4 is a carboxylic acid, then R.sup.4 in 3 is
preferably an ester that is hydrolyzed in the final step of the
scheme. If that ester is a t-butyl ester (i.e., if R.sup.4 is
CO.sub.2tBu), treatment with trifluoroacetic acid will give the
acid. The ester is preferably a t-butyl ester when the other
substituents in I are compatible with acidic conditions.
[0105] If R.sup.4 in product I is an ester, the desired ester may
be prepared by esterifying the corresponding acid or by having the
desired ester group already present in compound 2.
[0106] Scheme II 9
[0107] In scheme II above, the following abreviations are used: KF
is potassium fluoride; DMF is N,N-dimethylformamide; ArOH is
2,3,5,6-tetrafluorophenol; THF is tetahydrofuran; MeOH is methanol.
Commercially available bromoketone 4 (R.sup.4.dbd.CO.sub.2tBu) is
reacted with 2,3,5,6-tetrafluorophenol and potassium fluoride to
give phenoxy ketone 5. The ketone is then reduced with, for
example, sodium borohydride to give the alcohol 6, which is
hydrogenated by using, for example, palladium on carbon as catalyst
to give the amino alcohol 2 (R.sup.4.dbd.CO.sub.2tBu,
R.sup.5.dbd.CH.sub.2O-2,3,5,6-tetrafluorophenyl- ).
[0108] Scheme III 10
[0109] In Scheme III the folowing abreviatons are used: Z is a
benzyloxycarbonyl protecting group; MeOH is methanol; DCM is
dichloromethane; TFA is trifluoroacetic acid; DMAP is
4-dimethylaminopyridine; THF is tetrahydrofuran. Pyridone acid
derivatives I can be prepared in chiral form using the synthetic
sequence shown in Scheme III. The starting
(2-oxo-1,2-dihydro-pyridin-3-yl)-carbam- ic acid benzyl ester
(R.sup.2.dbd.H) is prepared using a procedure similar to that
described by Warner et al J. Med. Chem. 1994, 37(19), 3090-3099
Commercially available (R)-tert-butyl-2-hydroxybutyrate
(R.sup.3=ethyl) is treated with trifluoromethanesulphonic anhydride
and 2,6-lutidine in DCM to give the corresponding triflate.
Reaction of the triflate with the anion of
(2-oxo-1,2-dihydro-pyridin-3-yl)-carbamic acid benzyl ester
(prepared by deprotonation with sodium hydride in THF) gives the
N-alkylated pyridone. Removal of the benzyloxycarbonyl protecting
group using hydrogen and palladium on carbon gives the amine. This
is then reacted with an appropriate electrophile, triethylamine and
DMAP in DCM. For example if R.sup.1 is required to be RC.dbd.O (an
amide) then an appropriately substituted acid chloride may be used.
If R.sup.1 is required to be RS(.dbd.O).sub.2 (sulphonamide) then
an appropriately substituted sulfonyl chloride may be used. If
R.sup.1 is RO(C.dbd.O) (carbamate) then an appropriately
substituted chloroformate may be used. If R.sup.1 is RN(C.dbd.O)
(urea) then an appropriately substituted carbamoyl chloride or
isocyanate may be used. The other R.sup.1 groups may be prepared
accordingly. Acid 1 is then prepared by deprotection of the ester
by, for example, using trifluoroacetic acid. The acid is then
coupled to amino alcohol 2 (Scheme 1).
[0110] Therefore, another embodiment of this invention provides a
process for preparing a compound of formula I: 11
[0111] wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4, and R.sup.5, are
as defined in any of the embodiments herein, comprising:
[0112] (a) reacting a compound of formula (III): 12
[0113] wherein:
[0114] R.sup.9 is --NO.sub.2, --C(O)OR.sup.10, R.sup.6C(O)N(H)--,
R.sup.6SO.sub.2N(H)--, R.sup.6OC(O)N(H)--,
(R.sup.6).sub.2NC(O)N(H)--, R.sup.6C(O)C(O)N(H)--, R.sup.6N(H)--,
(R.sup.6).sub.2NC(O)C(O)N(H)--, or R.sup.6OC(O)(O)N(H)--;
[0115] R.sup.10 is independently hydrogen,
(C1-C12)-aliphatic-(C3-C10)-cyc- loaliphatic-, (C6-C10)-aryl-,
(C3-C10)-heterocyclyl-, (C5-C10)-heteroaryl-,
(C3-C10)-cycloaliphatic-(C1-C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)-aliphatic-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphat- ic-,
(C5-C10)-heteroaryl(C.sub.1-C12)-aliphatic-, wherein up to 3
aliphatic carbon atoms may be replaced with a group selected from
O, N(H), N(R), S, SO, and SO.sub.2; and wherein R.sup.10 is
optionally substituted with up to 6 substituents independently
selected from R; and
[0116] R, R.sup.2, R.sup.3 and R.sup.6 are as defined in any of the
embodiments of formula (I) herein;
[0117] with a compound of formula (IV): 13
[0118] wherein Y is either a carbonyl group or an OH group; and
[0119] R.sup.4 and R.sup.5 are as defined in any of the embodiments
of formula (I) herein;
[0120] in the presence of peptide coupling conditions and a
solvent;
[0121] provided that if Y is an OH group, then the process further
comprises (b) oxidizing the OH group to provide the compound of
formula (I); and
[0122] provided that if R.sup.9 is --NO.sub.2, --C(O)OR.sup.10, or
--CN, the process comprises the further step of converting the
--NO.sub.2, --C(O)OR.sup.10, or --CN into R.sup.6C(O)N(H)--,
R.sup.6SO.sub.2N (H)--, R.sup.6OC(O)N(H)--,
(R.sup.6).sub.2NC(O)N(H)--, R.sup.6C(O)C(O)N(H)--, R.sup.6N(H)--,
(R.sup.6).sub.2NC(O)C(O)N(H)--, or R.sup.6OC(O)C(O)N(H)--.
[0123] The coupling conditions may be any known to skilled
practitioners for forming peptidyl bonds. Preferred coupling
conditions are EDC/DMAP/HOBt. A preferred solvent in the above
embodiment is THF.
[0124] In a preferred embodiment, the compound of formula (III):
14
[0125] wherein R.sup.2, R.sup.3, and R.sup.9 are as defined
herein;
[0126] is prepared by a process comprising:
[0127] (c) reacting a compound of formula (V): 15
[0128] wherein R, R.sup.2, R.sup.3, and R.sup.9 are as defined
herein;
[0129] in a solvent in the presence of deprotecting conditions.
[0130] The deprotecting conditions will depend on the specific
protecting group (i.e., R.sup.10). For example, if R.sup.10 is
t-butyl, then preferred deprotecting conditions would include acid
hydrolysis. A preferred acid is TFA. A preferred solvent is DCM.
More preferably the solvent and the hydrolyzing conditions comprise
TFA and DCM. If R.sup.10 is methyl or ethyl, then preferred
deprotecting conditions would be basic (e.g., aqueous NaOH). If
R.sup.10 is benzyl, then the benzyl group could be removed by
hydrogenolysis.
[0131] In a preferred embodiment, the compound of formula (V):
16
[0132] wherein R.sup.2, R.sup.3, R.sup.9, and R.sup.10 are as
defined herein;
[0133] is prepared by a process comprising:
[0134] (d) reacting a compound of formula (VI): 17
[0135] wherein R.sup.2 and R.sup.9 are as defined herein;
[0136] with a compound of formula (VII): 18
[0137] wherein X is a suitable leaving group; and
[0138] R.sup.3 and R.sup.10 are as defined herein;
[0139] in the presence of a solvent and a base.
[0140] Preferably, X is --I, --Br, --Cl, --OH, an alkylsulfonate,
or an aryl sulfonate. When X is --OH, an appropriate leaving group
may be generated in situ (e.g., as in the Mitsunobu reaction).
Preferred sulfonates include --O-trifluoromethanesulfonate,
--O-methanesulfonate, --O-benzenesulfonate, --O-p-toluenesulfonate,
--O-m-nitrobenzenesulfonate- , and --O-p-nitrobenzenesulfonate.
Suitable leaving groups useful in the methods of this invention are
well known in the art. See, e.g., "March's Advanced Organic
Chemistry", 5.sup.th Ed., Ed.: Smith, M. B. and March, J., John
Wiley & Sons, New York (2001).
[0141] Any solvent that is compatible with the generation of anions
may be used. Preferred solvents include DMF, toluene, and THF.
[0142] Suitable bases include any that may remove a proton from the
hydroxy group in (V). Such bases include BuLi, LDA, LHMDS, and NaH.
Preferably, the base is NaH.
[0143] Another embodiment of this invention provides a process for
preparing a compound of formula (VIII): 19
[0144] wherein:
[0145] R.sup.2 is --CF.sub.3, --Cl, --OR.sup.7, --NO.sub.2,
--OCF.sub.3, --CN, or R.sup.8; and
[0146] R.sup.3, R.sup.8, R.sup.9, and R.sup.10 are as defined
herein;
[0147] comprising the step of (e) reacting a compound of formula
(IX): 20
[0148] wherein R.sup.2 and R.sup.9 are as defined herein;
[0149] with a compound of formula (VII): 21
[0150] wherein R.sup.3 and R.sup.10 are as defined herein; and
[0151] X is a suitable leaving group;
[0152] in the presence of a solvent and a base.
[0153] Preferably, X is --I, --Br, --Cl, --OH, an alkylsulfonate,
or an aryl sulfonate. When X is --OH, an appropriate leaving group
may be generated in situ (e.g., as in the Mitsunobu reaction).
Preferred sulfonates include --O-trifluoromethanesulfonate,
--O-methanesulfonate, --O-benzenesulfonate, --O-p-toluenesulfonate,
--O-m-nitrobenzenesulfonate- , and --O-p-nitrobenzenesulfonate.
[0154] Any solvent is compatible with the generation of anions may
be used. Such solvents include DMF, toluene, and THF. Preferably,
the solvent is THF.
[0155] Suitable bases include any that may remove a proton from the
hydroxy group in (V). Such bases include BuLi, LDA, LHMDS, and NaH.
Preferably, the base is NaH.
[0156] Another embodiment of this invention provides a process for
preparing a compound of formula (I): 22
[0157] wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4, and R.sup.5, are
as defined in any of the embodiments herein, comprising:
[0158] (a) reacting a compound of formula (VI or IX): 23
[0159] wherein:
[0160] R.sup.9 is --NO.sub.2, --C(O)OR.sup.10, --CN,
R.sup.6C(O)N(H)--, R.sup.6SO.sub.2N(H)--, R.sup.6OC(O)N(H)--,
(R.sup.6).sub.2NC(O)N(H)--, R.sup.6C(O)C(O)N(H)--, R.sup.6N(H)--,
(R.sup.6).sub.2NC(O)C(O)N(H)--, or R.sup.6OC(O)C(O)N(H)--; and
[0161] R.sup.2, R.sup.3 and R.sup.6 are as defined herein;
[0162] with a compound of formula (X): 24
[0163] wherein Y is either a carbonyl group or an OH group; and
[0164] R.sup.4 and R.sup.5 are as defined herein;
[0165] in the presence of any of the coupling conditions defined
herein and a solvent;
[0166] provided that if Y is an OH group, then the process further
comprises (b) oxidizing the OH group to provide the compound of
formula (I); and
[0167] provided that if R.sup.9 is --NO.sub.2, --C(O)OR.sup.10, or
--CN, the process comprises the further step of converting the
'NO.sub.2, --C(O)OR.sup.10, or --CN into R.sup.6(O)N(H)--,
R.sup.6SO.sub.2N(H)--, R.sup.6OC(O)N(H)--,
(R.sup.6).sub.2NC(O)N(H)--, R.sup.6C(O)C(O)N(H)--, R.sup.6N(H)--,
(R.sup.6).sub.2NC(O)C(O)N(H)--, or R.sup.6OC(O)C(O)N(H)--.
[0168] The compounds of this invention can be assayed for their
ability to inhibit the release of IL-1.beta., caspase activity, or
apoptosis directly. Assays for each of the activities are known in
the art. Selected assays are described below.
[0169] If pharmaceutically acceptable salts of the compounds of
this invention are utilized in these compositions, those salts are
preferably derived from inorganic or organic acids and bases.
Included among such acid salts are the following: acetate, adipate,
alginate, aspartate, benzoate, benzene sulfonate, bisulfate,
butyrate, citrate, camphorate, camphor sulfonate,
cyclopentanepropionate, digluconate, dodecylsulfate,
ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate,
hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide,
hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate,
methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate,
pamoate, pectinate, persulfate, 3-phenyl-propionate, picrate,
pivalate, propionate, succinate, tartrate, thiocyanate, tosylate
and undecanoate. Base salts include ammonium salts, alkali metal
salts, such as sodium and potassium salts, alkaline earth metal
salts, such as calcium and magnesium salts, salts with organic
bases, such as dicyclohexylamine salts, N-methyl-D-glucamine, and
salts with amino acids such as arginine, lysine, and so forth.
[0170] Also, the basic nitrogen-containing groups can be
quaternized with such agents as lower alkyl halides, such as
methyl, ethyl, propyl, and butyl chloride, bromides and iodides;
dialkyl sulfates, such as dimethyl, diethyl, dibutyl and diamyl
sulfates, long chain halides such as decyl, lauryl, myristyl and
stearyl chlorides, bromides and iodides, aralkyl halides, such as
benzyl and phenethyl bromides and others. Water or oil-soluble or
dispersible products are thereby obtained.
[0171] The compounds utilized in the compositions and methods of
this invention may also be modified by appending appropriate
functionalities to enhance selective biological properties. Such
modifications are known in the art and include those which increase
biological penetration into a given biological system (e.g., blood,
lymphatic system, central nervous system), increase oral
availability, increase solubility to allow administration by
injection, alter metabolism and alter rate of excretion.
[0172] Pharmaceutically acceptable carriers that may be used in
these compositions include, but are not limited to, ion exchangers,
alumina, aluminum stearate, lecithin, serum proteins, such as human
serum albumin, buffer substances such as phosphates, glycine,
sorbic acid, potassium sorbate, partial glyceride mixtures of
saturated vegetable fatty acids, water, salts or electrolytes, such
as protamine sulfate, disodium hydrogen phosphate, potassium
hydrogen phosphate, sodium chloride, zinc salts, colloidal silica,
magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based
substances, polyethylene glycol, sodium carboxymethylcellulose,
polyacrylates, waxes, polyethylene-polyoxypropyle- ne-block
polymers, polyethylene glycol and wool fat.
[0173] According to a preferred embodiment, the compositions of
this invention are formulated for pharmaceutical administration to
a mammal, preferably a human being.
[0174] Such pharmaceutical compositions of the present invention
may be administered orally, parenterally, by inhalation spray,
topically, rectally, nasally, buccally, vaginally or via an
implanted reservoir. The term "parenteral" as used herein includes
subcutaneous, intravenous, intramuscular, intra-articular,
intra-synovial, intrasternal, intrathecal, intrahepatic,
intralesional and intracranial injection or infusion techniques.
Preferably, the compositions are administered orally or
intravenously.
[0175] Sterile injectable forms of the compositions of this
invention may be aqueous or oleaginous suspension. These
suspensions may be formulated according to techniques known in the
art using suitable dispersing or wetting agents and suspending
agents. The sterile injectable preparation may also be a sterile
injectable solution or suspension in a non-toxic
parenterally-acceptable diluent or solvent, for example as a
solution in 1,3-butanediol. Among the acceptable vehicles and
solvents that may be employed are water, Ringer's solution and
isotonic sodium chloride solution. In addition, sterile, fixed oils
are conventionally employed as a solvent or suspending medium. For
this purpose, any bland fixed oil may be employed including
synthetic mono- or di-glycerides. Fatty acids, such as oleic acid
and its glyceride derivatives are useful in the preparation of
injectables, as are natural pharmaceutically-acceptable oils, such
as olive oil or castor oil, especially in their polyoxyethylated
versions. These oil solutions or suspensions may also contain a
long-chain alcohol diluent or dispersant, such as carboxymethyl
cellulose or similar dispersing agents which are commonly used in
the formulation of pharmaceutically acceptable dosage forms
including emulsions and suspensions. Other commonly used
surfactants, such as Tweens, Spans and other emulsifying agents or
bioavailability enhancers which are commonly used in the
manufacture of pharmaceutically acceptable solid, liquid, or other
dosage forms may also be used for the purposes of formulation.
[0176] The pharmaceutical compositions of this invention may be
orally administered in any orally acceptable dosage form including,
but not limited to, capsules, tablets, aqueous suspensions or
solutions. In the case of tablets for oral use, carriers which are
commonly used include lactose and corn starch. Lubricating agents,
such as magnesium stearate, are also typically added. For oral
administration in a capsule form, useful diluents include lactose
and dried corn starch. When aqueous suspensions are required for
oral use, the active ingredient is combined with emulsifying and
suspending agents. If desired, certain sweetening, flavoring or
coloring agents may also be added.
[0177] Alternatively, the pharmaceutical compositions of this
invention may be administered in the form of suppositories for
rectal administration. These can be prepared by mixing the agent
with a suitable non-irritating excipient which is solid at room
temperature but liquid at rectal temperature and therefore will
melt in the rectum to release the drug. Such materials include
cocoa butter, beeswax and polyethylene glycols.
[0178] The pharmaceutical compositions of this invention may also
be administered topically, especially when the target of treatment
includes areas or organs readily accessible by topical application,
including diseases of the eye, the skin, or the lower intestinal
tract. Suitable topical formulations are readily prepared for each
of these areas or organs.
[0179] Topical application for the lower intestinal tract can be
effected in a rectal suppository formulation (see above) or in a
suitable enema formulation. Topically-transdermal patches may also
be used.
[0180] For topical applications, the pharmaceutical compositions
may be formulated in a suitable ointment containing the active
component suspended or dissolved in one or more carriers. Carriers
for topical administration of the compounds of this invention
include, but are not limited to, mineral oil, liquid petrolatum,
white petrolatum, propylene glycol, polyoxyethylene,
polyoxypropylene compound, emulsifying wax and water.
Alternatively, the pharmaceutical compositions can be formulated in
a suitable lotion or cream containing the active components
suspended or dissolved in one or more pharmaceutically acceptable
carriers. Suitable carriers include, but are not limited to,
mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters
wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and
water.
[0181] For ophthalmic use, the pharmaceutical compositions may be
formulated as micronized suspensions in isotonic, pH adjusted
sterile saline, or, preferably, as solutions in isotonic, pH
adjusted sterile saline, either with our without a preservative
such as benzylalkonium chloride. Alternatively, for ophthalmic
uses, the pharmaceutical compositions may be formulated in an
ointment such as petrolatum.
[0182] The pharmaceutical compositions of this invention may also
be administered by nasal aerosol or inhalation. Such compositions
are prepared according to techniques well-known in the art of
pharmaceutical formulation and may be prepared as solutions in
saline, employing benzyl alcohol or other suitable preservatives,
absorption promoters to enhance bioavailability, fluorocarbons,
and/or other conventional solubilizing or dispersing agents.
[0183] The above-described compositions are particularly useful in
therapeutic applications relating to an IL-1 mediated disease, an
apoptosis mediated disease, an inflammatory disease, an autoimmune
disease, a destructive bone disorder, a proliferative disorder, an
infectious disease, a degenerative disease, a disease associated
with cell death, or various forms of liver disease. Such diseases
include those related to rheumatology and autoimmunity, such as
rheumatoid arthritis, osteoarthritis, osteoporosis, systemic lupus
erythematosus, scleroderma, chronic thyroiditis, Grave's disease,
myasthenia gravis, autoimmune neutropenia, autoimmune hemolytic
anemia, thrombocytopenia, juvenile rheumatoid arthritis, gout,
Behcet's syndrome, Still's syndrome, macrophage activation
syndrome, and sarcoidosis; auto-inflammatory syndromes, such as
cryopyrin-associated Periodic Syndromes, (including Muckle-Wells
syndrome, familial cold urticaria, chronic infantile neurological
cutaneous and articular syndrome (a.k.a. neonatal onset multisystem
inflammatory disease)), familial Mediterranean fever,
TNFR1-Associated Periodic Syndrome (TRAPS), Hyper-IgD periodic
fever Syndrome (HIDS), and Blau's syndrome; dermatology, such as
psoriasis, atopic dermatitis, scarring, alopecia, acne vulgaris,
and pemphigus; respiratory, such as asthma, adult respiratory
distress syndrome, cystic fibrosis, emphysema, chronic bronchitis,
chronic obstructive pulmonary disease, and idiopathic pulmonary
fibrosis; internal medicine, such as inflammatory peritonitis,
inflammatory bowel disease, Crohn's disease, ulcerative colitis,
autoimmune gastritis, H.pylori-associated gastric and duodenal
ulcer disease, diabetes, pancreatitis, glomerulonephritis, chronic
active hepatitis, excess dietary alcohol intake disease, renal
disease, polycystic kidney disease, burns, organ apoptosis after
burn injury, haemorrhagic shock, organ failure (e.g., hepatic
failure, acute renal failure, and acute respiratory failure), and
endometriosis; transplants, such as graft vs. host disease (GVHD)
and organ transplant rejection; oncology, such as leukemias and
related disorders, myelodysplastic syndrome, multiple
myeloma-related bone disorder, acute myelogenous leukemia, chronic
myelogenous leukemia, metastatic melanoma, Kaposi's sarcoma, and
multiple myeloma; cardiovascular, such as chronic heart disease,
acute heart disease, myocardial infarction, myocardial ischemia,
congestive heart failure, atherosclerosis, coronary artery bypass
graft (CABG), and acute coronary syndrome; the central and
peripheral nervous systems, such as Alzheimer's disease,
Parkinson's disease, Huntington's disease, Kennedy's disease, prion
disease, cerebral ischemia, epilepsy, spinal muscular atrophy,
amyotrophic lateral sclerosis, multiple sclerosis, HIV-related
encephalitis, traumatic brain injury, spinal cord injury,
neurological damage due to stroke, diabetic neuropathy, and acute
and chronic pain; ophthalomology, such as uveitis, retinal
disorders, diabetic retinopathy, glaucoma, and keratitis;
infectious diseases, such as viral mediated disease, sepsis, septic
shock, Shigellosis, hepatitis-B, hepatitis-C, hepatitis-G, yellow
fever, dengue fever, Japanese encephalitis, HIV infection,
tuberculosis, meningitis, Pseudomonas infection, and Acinetobacter
infection; and other diseases, such as aging. The compounds and
compositions are also useful in treating complications associated
with coronary artery bypass grafts. The amount of compound present
in the above-described compositions should be sufficient to cause a
detectable decrease in the severity of the disease or in caspase
activity and/or cell apoptosis, as measured by any of the assays
known in the art.
[0184] According to another embodiment, the compositions of this
invention may further comprise another therapeutic agent. Such
agents include, but are not limited to, thrombolytic agents such as
tissue plasminogen activator and streptokinase. When a second agent
is used, the second agent may be administered either as a separate
dosage form or as part of a single dosage form with the compounds
or compositions of this invention. Accordingly, a combined
preparation for simultaneous, separate, or sequential use is
provided by this invention.
[0185] Dosage levels of between about 0.01 and about 100 mg/kg body
weight per day, preferably between about 0.5 and about 75 mg/kg
body weight per day of the protease inhibitor compounds described
herein are useful in a monotherapy for the prevention and treatment
of a disease involving caspase activity and/or apoptosis.
[0186] Typically, the pharmaceutical compositions of this invention
will be administered from about 1 to about 5 times per day or
alternatively, as a continuous infusion. Such administration can be
used as a chronic or acute therapy. The amount of active ingredient
that may be combined with the carrier materials to produce a single
dosage form will vary depending upon the host treated and the
particular mode of administration. A typical preparation will
contain from about 5% to about 95% active compound (w/w).
Preferably, such preparations contain from about 20% to about 80%
active compound.
[0187] When the compositions of this invention comprise a
combination of a compound of formula I and one or more additional
therapeutic or prophylactic agents, both the compound and the
additional agent should be present at dosage levels of between
about 10 to 100%, and more preferably between about 10 to 80% of
the dosage normally administered in a monotherapy regimen.
[0188] It should also be understood that a specific dosage and
treatment regimen for any particular patient will depend upon a
variety of factors, including the activity of the specific compound
employed, the age, body weight, general health, sex, diet, time of
administration, rate of excretion, drug combination, and the
judgment of the treating physician and the severity of the
particular disease being treated. The amount of active ingredients
will also depend upon the particular compound and other therapeutic
agent, if present, in the composition.
[0189] In a preferred embodiment, the invention provides a method
of treating a mammal, having one of the aforementioned diseases,
comprising the step of administering to said mammal a
pharmaceutically acceptable composition described above. In this
embodiment, if the patient is also administered another therapeutic
agent or caspase inhibitor, it may be delivered together with the
compound of this invention in a single dosage form, or, as a
separate dosage form. When administered as a separate dosage form,
the other caspase inhibitor or agent may be administered prior to,
at the same time as, or following administration of a
pharmaceutically acceptable composition comprising a compound of
this invention.
[0190] In order that this invention be more fully understood, the
following preparative and testing examples are set forth. These
examples are for the purpose of illustration only and are not to be
construed as limiting the scope of the invention in any way.
EXAMPLE 1
(S,S)-3-[2-(3-Acetylamino-2-oxo-2H-pyridin-1-yl)-butyrylamino]-4-oxo-5-(2,-
3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0191] 25
Method A
(S)-2-(3-Benzyloxycarbonylamino-2-oxo-2H-pyridin-1-yl)-butyric acid
tert-butyl ester
[0192] 26
[0193] To a cooled (0.degree. C.) solution of (R)-tert-butyl
hydroxybutyrate (1.03 g, 6.43 mmol) in dichloromethane (25 mL), was
slowly added 2,6-lutidine (1.38 g, 12.9 mmol) and then
trifluoromethanesulfonic anhydride (3.45 g, 12.2 mmol). The
resulting mixture was stirred at 0.degree. C. for 1 hour, then
partitioned between tert-butylmethyl ether (150 mL) and an aqueous
solution of 1M HCl (30 mL). The organic layer was washed with brine
(30 mL), dried (sodium sulfate), filtered and concentrated to
afford the triflate as a light brown oil.
[0194] To a solution of (2-oxo-1,2-dihydro-pyridin-3-yl)-carbamic
acid benzyl ester (P. Warner et al., J. Med. Chem., 37, 19, 1994,
3090-3099)(1.73 g, 7.07 mmol) in dry THF (60 mL) was added sodium
hydride (60% dispersion, 257 mg, 6.43 mmol) and the solution was
stirred at room temperature for 45 minutes. The reaction mixture
was then slowly transferred with a canula onto a solution of the
triflate prepared above in THF (3 mL). The reaction mixture was
stirred at room temperature for 90 minutes and quenched with
aqueous ammonium chloride (10 mL). Most of the solvent was
evaporated and the residue was partitioned between EtOAc and
saturated aqueous NH4Cl. The organic layer was washed with brine
(30 mL), dried (MgSO.sub.4), filtered and evaporated. The residue
was purified by flash chromatography (10% ethyl acetate/hexane) to
afford the title compound as a colourless oil (2.48 g, 100%):
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 0.92(3H, t), 1.45(9H, s),
1.94(1H, m), 2.25(1H, m), 5.23 (2H, s), 5.47 (1H, dd), 6.32 (1H,
t), 7.01 (1H, d), 7.32-7.43 (5H, m), 7.92 (1H, s), 8.06 (1H, br
d).
Method B
(S)-2-(3-Amino-2-oxo-2H-pyridin-1-yl)-butyric acid tert-butyl
ester
[0195] 27
[0196] To a solution of
(S)-2-(3-Benzyloxycarbonylamino-2-oxo-2H-pyridin-1- -yl)-butyric
acid tert-butyl ester (2.48 g, 6.43 mmol) in a mixture of MeOH (15
mL) and EtOAc (15 mL) was added 10% Pd/C (250 mg). The mixture was
degassed and stirred at room temperature for 90 minutes under an
atmosphere of hydrogen (balloon pressure). The reaction mixture was
filtered through a short pad of silica which was then flushed with
MeOH. The combined filtrates were evaporated under reduced pressure
to afford the title compound as a white solid (1.62 g, 100%);
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 0.91 (3H, t), 1.44(9H,
s), 1.91(1H, m), 2.21(1H, m), 4.24 (2H, br s), 5.50 (1H, dd), 6.11
(1H, t), 6.53 (1H, d), 6.77 (1H, d).
Method C
(S)-2-(3-Acetylamino-2-oxo-2H-pyridin-1-yl)-butyric acid tert-butyl
ester
[0197] 28
[0198] To a-cooled (0.degree. C.) solution of
(S)-2-(3-Amino-2-oxo-2H-pyri- din-1-yl)-butyric acid tert-butyl
ester (500 mg, 1.98 mmol) in dichloromethane (5 mL) was added
triethylamine (220 mg, 2.18 mmol) followed by acetic anhydride (202
mg, 1.98 mmol). The reaction mixture was stirred at room
temperature for 12 hours and then partitioned between EtOAc and
aqueous 1M HCl. The organic layer was washed with saturated aqueous
NaHCO.sub.3, brine (30 mL), dried (MgSO.sub.4), filtered and
evaporated. The residue was purified by flash chromatography (40%
ethyl acetate/hexane) to afford the title compound as a colourless
oil (569 mg, 97%): .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 0.87
(3H, t), 1.40(9H, s), 1.91(1H, m), 2.13 (3H, s), 2.19(1H, m), 5.38
(1H, dd), 6.26 (1H, t), 6.99 (1H, d), 8.33 (1H, d), 8.43 (1H, br
s).
Method D
(S)-2-(3-Acetylamino-2-oxo-2H-pyridin-1-yl)-butyric acid
[0199] 29
[0200] A solution of
(S)-2-(3-Acetylamino-2-oxo-2H-pyridin-1-yl)-butyric acid tert-butyl
ester (569 mg, 1.93 mmol) in dichloromethane (5 mL) was cooled to
0.degree. C. Trifluoroacetic acid (5 ml) was added and the
resulting mixture allowed to warm to room temperature and stir for
2 hours. The mixture was then concentrated under reduced pressure
and the residue redisolved in dichloromethane. This process was
repeated several times in order to remove excess trifluoroacetic
acid. The resulting solid was slurried in diethyl ether, filtered
and washed with more diethyl ether. The solid was then dried to
constant weight under vacuum. This gave the title product as a
white solid (327 mg, 71%); .sup.1H NMR (400 MHz, d6-DMSO) .delta.
0.78 (3H, t), 2.02-2.17 (5H, m), 4.98 (1H, dd), 6.29 (1H, t), 7.35
(1H, d), 8.21 (1H, d), 9.30 (1H, s), 13.07 (1H, vbr s).
Method E
(S,S)-3-[2-(3-Acetylamino-2-oxo-2H-pyridin-1-yl)-butyrylamino]-4-hydroxy-5-
-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid tert-butyl ester
[0201] 30
[0202] A stirred mixture of
(S)-2-(3-Acetylamino-2-oxo-2H-pyridin-1-yl)-bu- tyric acid (100 mg,
0.42 mmol), 3-amino-5-(2,3,5,6-tetrafluorophenoxy)-4-h-
ydroxy-pentanoic acid tert-butyl ester (163 mg, 0.462 mmol), HOBt
(62 mg, 0.462 mmol), DMAP (56 mg, 0.462 mmol)and THF (5 mL) was
cooled to 0.degree. C. then EDC (89 mg, 0.462 mmol) was added. The
mixture was allowed to warm to room temperature during 16 h then
concentrated under reduced pressure. The residue was purified by
flash chromatography (50-50% ethyl acetate/hexane) to afford the
title compound as a white foam (221 mg, 92%); .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 0.88-0.93 (3H, m), 1.37-1.38 (9H, 2s),
1.86-1.96 (1H, m), 2.15-2.25 (4H, m), 2.55-2.71 (2H, m), 3.70-4.64
(5H, m), 5.30-5.39 (1H, m), 6.30-6.35 (1H, m), 6.75-6.86 (1H, m),
7.17-7.31 (2H, m), 8.31-8.47 (2H, m).
Method F
(S,S)-3-[2-(3-Acetylamino-2-oxo-2H-pyridin-1-yl)-butyrylamino]-4-oxo-5-(2,-
3,5,6-tetrafluoro-phenoxy)-pentanoic acid tert-butyl ester
[0203] 31
[0204] A stirred solution of
(S,S)-3-[2-(3-Acetylamino-2-oxo-2H-pyridin-1--
yl)-butyrylamino]-4-hydroxy-5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic
acid tert-butyl ester (221 mg, 0.385 mmol) in anhydrous DCM (10 mL)
was treated with
1,1,1-triacetoxy-1,1-dihydro-1,2-benziodoxol-3(1H)-one (212 mg, 0.5
mmol) at 0.degree. C. The resulting mixture was kept at 0.degree.
C. for 2 hr, diluted with ethyl acetate, then poured into a 1:1
mixture of saturated aqueous sodium hydrogen carbonate and
saturated aqueous sodium thiosulfate. The organic layer was removed
and the aqueous layer re-extracted with ethyl acetate. The combined
organic extracts were dried (Magnesium sulfate) and concentrated.
The residue was purified by flash chromatography (50-50% ethyl
acetate/hexane) to afford the title compound as a white solid (187
mg, 85%); .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 0.93 (3H, t),
1.36 (3H, s), 1.95 (1H, m), 2.21 (3H, s), 2.25 (1H, m), 2.73 (2H,
dd), 2.89 (1H, dd), 4.91 (1H, m), 5.04-5.17 (2H, m), 5.47 (1H, m),
6.34 (1H, t), 6.80 (1H, m), 7.19 (1H, m), 7.68 (1H, d), 8.36-8.41
(2H, m).
Method G
(S,S)-3-[2-(3-Acetylamino-2-oxo-2H-pyridin-1-yl)-butyrylamino]-4-oxo-5-(2,-
3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0205] 32
[0206] A solution of
(S,S)-3-[2-(3-Acetylamino-2-oxo-2H-pyridin-1-yl)-buty-
rylamino]-4-oxo-5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
tert-butyl ester (187 mg, 0.327 mmol) in dichloromethane (5 mL) was
cooled to 0.degree. C. Trifluoroacetic acid (5 ml) was added and
the resulting mixture allowed to warm to room temperature and stir
for 2 hours. The mixture was then concentrated under reduced
pressure and the residue redisolved in dichloromethane. This
process was repeated several times in order to remove excess
trifluoroacetic acid. The resulting solid was slurried in diethyl
ether, filtered and washed with more diethyl ether. The solid was
then dried to constant weight under vacuum. This gave the title
product as a white solid (138 mg, 82%); .sup.1H NMR (400 MHz,
d6-DMSO) .delta. 0.78 (3H, t), 1.87-2.13 (5H, m), 2.56-2.78 (2H,
m), 4.62 (1H, m), 5.18-5.29 (2H, m), 5.40 (1H, m), 6.28 (1H, t),
7.37 (1H, d), 7.53-7.66 (1H, m), 8.17-8.21 (1H, m), 8.92 (1H, d),
9.21 (1H, s), 12.51 (1H, br s); .sup.19F NMR (376 MHz, d6-DMSO,
proton-decoupled) .delta. -156.9, -141.1; M+H 516.2, M-H 514.2.
EXAMPLE 2
(S,S)-4-Oxo-3-[2-(2-oxo-3-propionylamino-2H-pyridin-1-yl)-butyrylamino]-5--
(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0207] 33
[0208] Prepared from (S)-2-(3-Amino-2-oxo-2H-pyridin-1-yl)-butyric
acid tert-butyl ester and propionic anhydride according to methods
C-G; white solid; IR (solid) 1584, 1642, 1662, 1717, 1749 cm-1;
.sup.1H NMR (400 MHz, d6-DMSO) .delta. 0.78 (3H, t), 1.04 (3H, t),
1.88-2.11 (2H, m), 2.43 (2H, q), 2.59 (1H, d), 2.75 (1H, dd), 4.61
(1H, m), 5.18-5.29 (2H, 2dd), 5.40 (1H, m), 6.29 (1H, t), 7.37 (1H,
d), 7.58 (1H, m), 8.22 (1H, d), 8.91 (1H, d), 9.08 (1H, s), 12.50
(1H, br s); .sup.19F NMR (376 MHz, d6-DMSO, proton-decoupled)
.delta. -140.6, -140.8, -141.1, -156.8, -157.0; M+H 530.2, M-H
528.3.
EXAMPLE 3
(S,S)-3-[2-(3-Butyrylamino-2-oxo-2H-pyridin-1-yl)
butyrylamino]-4-oxo-5-(2- ,3,5,6-tetrafluoro-phenoxy)-pentanoic
acid
[0209] 34
[0210] Prepared from (S)-2-(3-Amino-2-oxo-2H-pyridin-1-yl)-butyric
acid tert-butyl ester and butyryl chloride according to methods
C-G; beige solid; IR (solid) 1659, 1645, 1509, 1490 cm-.sup.1;
.sup.1H NMR (400 MHz, d6-DMSO) .delta. 0.76-0.80 (3H, m), 0.88 (3H,
t), 1.53-1.58 (2H, m), 1.88-1.93 (1H, m), 2.01-2.09 (1H, m),
2.37-2.41 (2H, m), 2.59 (1H, dd), 2.70-2.81 (1H, m), 4.59-4.63 (1H,
m), 5.20-5.25 (2H, m), 5.38-5.50 (1H, 2.times.m), 7.36-7.38 (1H,
m), 7.55-7.61 (1H, m), 8.21-8.23 (1H, m), 8.61-8.92 (1H,
3.times.d), 9.06-9.10 (1H, m), 12.49 (1H, br s); .sup.19F NMR (376
MHz, d6-DMSO, proton-decoupled) .delta. -140.6, -141.1, -156.9,
-157.0;M+H 544.3, M-H 542.3.
EXAMPLE 4
(S,S)-3-{2-[3-(Cyclopropanecarbonyl-amino)-2-oxo-2H-pyridin-1-yl]-butyryla-
mino}-4-oxo-5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0211] 35
[0212] Prepared from (S)-2-(3-Amino-2-oxo-2H-pyridin-1-yl)-butyric
acid tert-butyl ester and cyclopropanecarbonyl chloride according
to methods C-G; white solid; .sup.1H NMR (400 MHz, d6-DMSO) .delta.
0.74-0.82 (7H, m), 1.93 (1H, m), 2.07 (1H, m), 2.17 (1H, m), 2.59
(1H, d), 2.75 (1H, dd), 4.62 (1H, m), 5.19-5.30 (2H, 2dd), 5.41
(1H, m), 6.27 (1H, t), 7.37 (1H, d), 7.57 (1H, m), 8.17 (1H, d),
8.92 (1H, d), 9.49 (1H, s), 12.51 (1H, br s); M+H 542.2, M-H
540.3.
EXAMPLE 5
(S,S)-3-[2-(3-Isobutyrylamino-2-oxo-2H-pyridin-1-yl)-butyrylamino]-4-oxo-5-
-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0213] 36
[0214] Prepared from (S)-2-(3-Amino-2-oxo-2H-pyridin-1-yl)-butyric
acid tert-butyl ester and isobutyryl chloride according to methods
C-G; white solid; IR (solid) 1664, 1517, 1491 cm-.sup.1; .sup.1H
NMR (400 MHz, d6-DMSO) .delta. 1.75-1.85 (3H, m), 1.05 (6H, d),
1.9-2.1 (2H, m), 2.6-2.9 (3H, m), 4.55-4.62 (1H, m), 5.2-5.35 (2H,
m), 5.4-5.43 (1H, m), 6.25 (1H, t), 7.4-7.45 (1H, m), 7.6-7.7 (1H,
m), 8.2-8.24 (1H, m), 8.8-9.0 (2H, m); M+H 544.3, M-H 542.3.
EXAMPLE 6
(S,S)-3-{2-[3-(2-Methoxy-acetylamino)-2-oxo-2H-pyridin-1-yl]-butyrylamino}-
-4-oxo-5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0215] 37
[0216] Prepared from (S)-2-(3-Amino-2-oxo-2H-pyridin-1-yl)-butyric
acid tert-butyl ester and methoxyacetyl chloride according to
methods C-G; pink solid; .sup.1H NMR (400 MHz, d6-DMSO) .delta.
0.75-0.80 (3H, m), 1.88-1.97 (1H, m), 2.02-2.10 (1H, m), 2.56-2.63
(1H, m), 2.72-2.79 (1H, m), 3.37-3.40 (3H, m), 4.00-4.03 (2H, m),
4.53-4.65 (1H, m), 5.13-5.46 (3H, m), 6.32-6.35 (1H, m), 7.39-7.45
(1H, m), 7.51-7.66 (1H, m), 8.21-8.26 (1H, m), 8.92-8.98 (1H, m),
9.12-9.17 (1H, m), 12.51 (1H, br s); M+H 546.2, M-H 544.2.
EXAMPLE 7
(S,S)-3-(2-{3-[(Furan-2-carbonyl)-amino]-2-oxo-2H-pyridin-1-yl}-butyrylami-
no)-4-oxo-5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0217] 38
[0218] Prepared from (S)-2-(3-Amino-2-oxo-2H-pyridin-1-yl)-butyric
acid tert-butyl ester and 2-furoyl chloride according to methods
C-G; white solid; .sup.1H NMR (400 MHz, d6-DMSO) .delta. 0.81 (3H,
m), 1.95 (1H, m), 2.09 (1H, m), 2.60 (1H, dd), 2.77 (1H, dd), 4.61
(1H, m), 5.19-5.29 (2H, m), 5.42 (1H, m), 6.39 (1H, t), 6.74 (1H,
m), 7.30 (1H, m), 7.46-7.58 (2H, m), 7.95 (1H, m), 8.27 (1H, d),
8.98 (1Hd), 9.16 (1H, s), 12.50 (1H, br s); M+H 568.3, M-H
566.3.
EXAMPLE 8
(S,S)-3-(2-{3-[(Furan-3-carbonyl)-amino]-2-oxo-2H-pyridin-1-yl}-butyrylami-
no)-4-oxo-5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0219] 39
[0220] Prepared from (S)-2-(3-Amino-2-oxo-2H-pyridin-1-yl)-butyric
acid tert-butyl ester and 3-furoyl chloride according to methods
C-G; off-white solid; IR (solid) 1748, 1711, 1663, 1640, 1583,
1517, 1488 cm-.sup.1; .sup.1H NMR (400 MHz, d6-DMSO) .delta. 0.80
(3H, m), 1.90-2.20 (2H, m), 2.60-2.90 (2H, m), 4.65 (1H, m),
5.10-5.60 (3H, m), 6.40 (1H, t), 6.95 (1H, m), 7.40-7.65 (2H, m),
7.85 (1H, s), 8.20 (1H, m), 8.50 (1H, m), 8.90-9.20 (2H, m);
.sup.19F NMR (376 MHz, d6-DMSO, proton-decoupled) .delta. -141.0,
-156.8; M+H 568.2, M-H 566.3.
EXAMPLE 9
(S,S)-4-Oxo-3-(2-{2-oxo-3-[(pyridine-3-carbonyl)-amino]-2H-pyridin-1-yl)-b-
utyrylamino)-5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0221] 40
[0222] Prepared from (S)-2-(3-Amino-2-oxo-2H-pyridin-1-yl)-butyric
acid tert-butyl ester and 3-pyridinecarbonyl chloride according to
methods C-G (isolated as a TFA salt); yellow solid; IR (solid)
1745, 1678, 1650, 1517, 1488; .sup.1H NMR (400 MHz, d6-DMSO)
.delta. 0.80 (3H, m), 1.90-2.30 (2H, m), 2.50-2.90 (2H, m), 4.65
(1H, m), 5.10-5.65 (3H, m), 6.45 (1H, t), 7.40-7.80 (3H, m),
8.10-8.40 (2H, m), 8.85 (1H, s), 8.90-9.20 (2H, m), 9.65 (1H, m);
.sup.19F NMR (376 MHz, d6-DMSO, proton-decoupled) .delta. -141.0,
-156.8; M+H 579.2, M-H 577.3.
EXAMPLE 10
(S,S)-3-(2-{3-[(Isothiazole-3-carbonyl)-amino]-2-oxo-2H-pyridin-1-yl}-buty-
rylamino)-4-oxo-5-(2,3,5,6-tetrafluorophenoxy)-pentanoic acid
[0223] 41
[0224] Prepared from (S)-2-(3-Amino-2-oxo-2H-pyridin-1-yl)-butyric
acid tert-butyl ester and 3-isothiazolecarbonyl chloride according
to methods C-G; pink solid; IR (solid) 1678, 1649, 1516, 1493
cm-.sup.1; .sup.1H NMR (400 MHz, d6-DMSO) .delta. 0.85 (3H, m),
1.85-2.30 (2H, m), 2.50-2.90 (2H, m), 4.20-4.70 (1H, 2m), 5.10-5.60
(3H, m), 6.45 (1H, t), 7.40-7.70 (2H, m), 7.95 (1H, m), 8.40 (1H,
d), 8.95-9.15 (1H, 2m), 9.30 (1H, d), 10.00 (1H, 2s); .sup.19F NMR
(376 MHz, d6-DMSO, proton-decoupled) .delta. -141.0, -156.9; M+H
585.1, M-H 583.2.
EXAMPLE 11
(S,S)-3-[2-(3-Benzoylamino-2-oxo-2H-pyridin-1-yl)-butyrylamino]-4-oxo-5-(2-
,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0225] 42
[0226] Prepared from (S)-2-(3-Amino-2-oxo-2H-pyridin-1-yl)-butyric
acid tert-butyl ester and benzoyl chloride according to methods
C-G; pink solid; IR (solid) 1645, 1509, 1490 cm-.sup.1; .sup.1H NMR
(400 MHz, d6-DMSO) .delta. 0.79-0.85 (3H, m), 1.95-1.99 (1H, m),
2.06-2.10 (1H, m), 2.60 (1H, dd), 2.77 (1H, dd), 4.59-4.63 (1H, m),
5.25 (2H, m), 5.42-5.55 (1H, m), 6.38-6.42 (1H, m), 7.51-7.62 (5H,
m), 7.89-7.91 (2H, m), 8.27-8.31 (1H, m), 8.69-8.99 (1H, m), 9.28
(1H, m); .sup.19F NMR (376 MHz, d6-DMSO, proton-decoupled) .delta.
-140.6, -141.0, -156.9, -157.0; M+H 578.2, M-H 576.2.
EXAMPLE 12
(S,S)-4-Oxo-3-[2-(2-oxo-3-phenylacetylamino-2H-pyridin-1-yl)-butyrylamino]-
-5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0227] 43
[0228] Prepared from (S)-2-(3-Amino-2-oxo-2H-pyridin-1-yl)-butyric
acid tert-butyl ester and phenylacetyl chloride according to
methods C-G; pink solid; IR (solid) 1659, 1635, 1519 cm-.sup.1;
.sup.1H NMR (400 MHz, d6-DMSO) .delta. 0.77 (3H, t), 1.85-1.96 (1H,
m), 2.03-2.07 (1H, m), 2.59 (1H, dd), 2.71-2.77 (1H, m), 3.79 (2H,
s), 4.61-4.66 (1H, m), 5.16-5.29 (2H, m), 5.35-5.44 (1H, m), 6.28
(1H, t), 7.24-7.39 (6H, m), 7:52-7.67 (1H, m), 8.19-8.21 (1H, m),
8.61-8.92 (1H, m), 9.28 (1H, br s); .sup.19F NMR (376 MHz, d6-DMSO,
proton-decoupled) .delta. -140.6, -141.0, -156.90, -157.0; M+H
592.2, M-H 590.2.
EXAMPLE 13
(S,S)-3-[2-(3-Acetylamino-2-oxo-5-trifluoromethyl-2H-pyridin-1-yl)-butyryl-
amino]-4-oxo-5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0229] 44
[0230] Prepared from
(2-oxo-5-trifluoromethyl-1,2-dihydro-pyridin-3-yl)-ca- rbamic acid
benzyl ester according to methods A-G; white solid; IR (solid)
1659, 1514 cm-.sup.1; .sup.1H NMR (400 MHz, d6-DMSO) .delta. 0.79
(3H, t), 2.07-2.33 (5H, m), 2.59-2.79 (2H, m), 4.59-4.63 (1H, m),
5.18-5.29 (2H, m), 5.41-5.45 (1H, m), 7.55-7.62 (1H, m), 7.89 (1H,
s), 8.41-8.43 (1H, m), 9.04 (1H, d), 9.61-9.63 (1H, m); .sup.19F
NMR (376 MHz, d6-DMSO, proton-decoupled) .delta. -61.4, -140.7,
-141.1, -156.8-156.9, -157.02, -157.1; M+H 584.2, M-H 582.2.
EXAMPLE 14
(S,S)-3-{2-[3-(3-Ethyl-ureido)-2-oxo-2H-pyridin-1-yl]-butyrylamino}-4-oxo--
5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0231] 45
[0232] Prepared from (S)-2-(3-Amino-2-oxo-2H-pyridin-1-yl)-butyric
acid tert-butyl ester and ethyl isocyanate according to methods
C-G; pink solid; IR (solid) 1664, 1645, 1550, 1493, 1208 cm.sup.1;
.sup.1H NMR (400 MHz, d6-DMSO) .delta. 0.80 (3H, t), 1.05 (3H, t),
1.80-2.20 (2H, m), 2.50-2.85 (2H, m), 3.15 (2H, m), 4.65 (1H, m),
5.25 (2H, dd), 5.40 (1H, m), 6.25 (1H, t), 7.15 (1H, s), 7.25 (1H,
d), 7.60 (1H, m), 8.05 (1H, m), 8.20 (1H, s), 8.95 (1H, d);
.sup.19F NMR (376 MHz, d6-DMSO, proton-decoupled) .delta. -141.1,
-156.9; M+H 545.2, M-H 543.2.
EXAMPLE 15
(S,S)-3-{2-[3-(3,3-Diethyl-ureido)-2-oxo-2H-pyridin-1-yl]-butyrylamino}-4--
oxo-5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0233] 46
Method H
[0234] To a cooled (0.degree. C.) solution of
(S)-2-(3-Amino-2-oxo-2H-pyri- din-1-yl)-butyric acid tert-butyl
ester (400 mg, 1.59 mmol) in dichloroethane (3 mL) was added
triethylamine (0.254 mL, 1.82 mmol). This solution was added
dropwise to a solution of diphosgene (0.11 mL, 0.91 mmol) in
dichloroethane (7 mL) at 0.degree. C. over 10 minutes. The reaction
mixture was stirred at room temperature for 90 minutes and then
partitioned between EtOAc and aqueous IM HCl. The organic layer was
washed with brine, dried (MgSO.sub.4), filtered and evaporated to
afford the isocyanate as a brown oil.
[0235] To a cooled (0.degree. C.) solution of the isocyanate
prepared above (244 mg, 0.79 mmol) in dichloroethane (4 mL) was
added triethylamine (0.122 mL, 0.87 mmol) followed by diethylamine
(0.082 mL, 0.79 mmol). The reaction mixture was stirred at room
temperature for 3 hours and then partitioned between EtOAc and
aqueous 1M HCl. The organic layer was washed with brine, dried
(MgSO.sub.4), filtered and evaporated to afford a brown oily
residue which was purified by flash column chromatography (50%
ethyl acetate/hexane) to afford the diethylurea as a colourless
oil.
[0236] This intermediate was involved in the sequence described in
methods D-G to afford the title compound; pink solid; IR (solid)
1640, 1512, 1213 cm-1; .sup.1H NMR (400 MHz, d6-DMSO) .delta.
0.75-0.95 (3H, m), 1.10-1.40 (6H, m), 1.90-2.25 (2H, m), 2.60-2.90
(2H, m), 3.30-3.50 (4H, m), 4.75 (1H, m), 5.10-5.60 (3H, m), 6.35
(1H, t), 7.30 (1H, m), 7.75 (1H, m), 7.80 (1H, m), 8.05 (1H, m),
8.95-9.05 (1H, m); .sup.19F NMR (376 MHz, d6-DMSO,
proton-decoupled) .delta. -141.0, -156.9; M+H 573.3, M-H 571.2.
EXAMPLE 16
(S,S)-4-Oxo-3-(2-{2-oxo-3-[(pyrrolidine-1-carbonyl)-amino]-2H-pyridin-1-yl-
}-butyrylamino)-5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0237] 47
[0238] Prepared from (S)-2-(3-Amino-2-oxo-2H-pyridin-1-yl)-butyric
acid tert-butyl ester and pyrrolidine according to methods H, D-G;
pink solid; IR (solid) 1650, 1593, 1512, 1489, 1208 cm-1; .sup.1H
NMR (400 MHz, d6-DMSO) .delta. 0.80 (3H, m), 1.80-2.20 (6H, m),
2.60-2.90 (2H, m), 3.30-3.50 (4H, m), 4.60-4.75 (1H, m), 5.10-5.50
(3H, m), 6.30 (1H, t), 7.35 (1H, m), 7.50-7.75 (2H, m), 8.00 (1H,
m), 8.85-8.95 (1H, m); .sup.19F NMR (376 MHz, d6-DMSO,
proton-decoupled) .delta. -141.1, -156.9; M+H 571.3, M-H 569.3.
EXAMPLE 17
(S,S)-3-[2-(3-Methoxycarbonylamino-2-oxo-2H-pyridin-1-yl)-butyrylamino]-4--
oxo-5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0239] 48
[0240] Prepared from (S)-2-(3-Amino-2-oxo-2H-pyridin-1-yl)-butyric
acid tert-butyl ester and methyl chloroformate according to methods
C-G; pink solid; IR (solid) 1644, 1661, 1709 cm-1; .sup.1H NMR (400
MHz, d6-DMSO) .delta. 0.81 (3H, m), 1.95 (1H, m), 2.09 (1H, m),
2.50-2.98 (2H, m), 3.70 (3H, s), 4.20-5.50 (4H, m), 6.31 (1H, m),
7.40 (1H, m), 7.59 (1H, m), 7.82 (1H, m), 8.20 (1H, s), 8.55-9.00
(1H, d); .sup.19F NMR (376 MHz, d6-DMSO, proton-decoupled) .delta.
-140.6, -141.0, -141.1, -156.80, -156.9, -157.0, -157.1; M+H 532.3,
M-H 530.3.
EXAMPLE 18
(S,S)-3-[2-(3-Ethanesulfonylamino-2-oxo-2H-pyridin-1-yl)-butyrylamino]-4-o-
xo-5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0241] 49
[0242] Prepared from (S)-2-(3-Amino-2-oxo-2H-pyridin-1-yl)-butyric
acid tert-butyl ester and ethanesulfonyl chloride according to
methods C-G; pink solid; .sup.1H NMR (400 MHz, d6-DMSO) .delta.
0.74-0.82 (3H, m), 1.17-1.25 (3H, m), 1.85-2.10 (2H, m), 2.54-2.79
(2H, m), 3.09-3.15 (2H, m), 4.58-4.68 (1H, m), 5.13-5.38 (2H, m),
6.26-6.31 (1H, m), 7.34-7.38 (1H, m), 7.51-7.73 (2H, m), 8.72-8.76
(1H, m), 8.89-8.97 (1H, m), 12.51 (1H, br s); M+H 566.2, M-H
564.2.
EXAMPLE 19
(S,S)-4-Oxo-3-{2-[2-oxo-3-(propane-1-sulfonylamino)-2H-pyridin-1-yl]-butyr-
ylamino}-5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0243] 50
[0244] Prepared from (S)-2-(3-Amino-2-oxo-2H-pyridin-1-yl)-butyric
acid tert-butyl ester and propanesulfonyl chloride according to
methods C-G; pink solid; .sup.1H NMR (400 MHz, d6-DMSO) .delta.
0.74-0.82 (3H, m), 0.88-0.94 (3H, m), 1.63-1.74 (2H, m), 1.85-2.10
(2H, m), 2.56-2.79 (2H, m), 3.06-3.13 (2H, m), 4.58-4.68 (1H, m),
5.13-5.40 (2H, m), 6.26-6.31 (1H, m), 7.34-7.37 (1H, m), 7.50-7.62
(2H, m), 8.71-8.75 (1H, m), 8.90-8.97 (1H, m), 12.53 (1H, br s);
M+H 580.3, M-H 578.3.
EXAMPLE 20
(S,S)-4-Oxo-3-{2-[2-oxo-3-(propane-2-sulfonylamino)-2H-pyridin-1-yl]-butyr-
ylamino}-5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0245] 51
[0246] Prepared from (S)-2-(3-Amino-2-oxo-2H-pyridin-1-yl)-butyric
acid tert-butyl ester and isopropylsulfonyl chloride using methods
similar to C-G; pink solid; IR (solid) 1645, 1518 cm-.sup.1;
.sup.1H NMR (400 MHz, d6-DMSO) .delta. 1.7-1.8 (3H, m), 1.18-1.25
(6H, m), 1.85-2.05 (2H, m), 2.55-2.8 (2H, m), 3.2-3.3 (1H, m),
4.52-4.62 (1H, m), 5.15-5.32 (3H, m), 5.4-5.43 (1H, m), 6.25 (1H,
t), 7.3-7.35 (1H, m), 7.45-7.6 (2H, m), 8.6-8.7 (1H, m), 8.9-9.0
(1H, m); M+H 580.2, M-H 578.2.
EXAMPLE 21
(S,S)-3-2-(3-Benzenesulfonylamino-2-oxo-2H-pyridin-1-yl)-butyrylamino]-4-o-
xo-5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0247] 52
[0248] Prepared from (S)-2-(3-Amino-2-oxo-2H-pyridin-1-yl)-butyric
acid tert-butyl ester and benzenesulfonyl chloride according to
methods C-G; pink solid; .sup.1H NMR (400 MHz, d6-DMSO) .delta.
0.55-0.66 (3H, m), 1.72-1.84 (1H, m), 1.91-2.01 (1H, m), 2.53-2.61
(1H, m), 2.68-2.76 (1H, m), 4.54-4.63 (1H, m), 5.06-5.32 (2H, m),
6.20-6.25 (1H, m), 6.98-7.86 (9H, m), 8.84-8.90 (1H, m), 9.40-9.45
(1H, m), 12.51 (1H, br s); M+H 614.1, M-H 612.1.
EXAMPLE 22
(S,S)-3-[2-(3-Ethanesulfonylamino-2-oxo-5-trifluoromethyl-2H-pyridin-1-yl)-
-butyrylamino]-4-oxo-5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic
acid
[0249] 53
[0250] Prepared from
(2-oxo-5-trifluoromethyl-1,2-dihydro-pyridin-3-yl)-ca- rbamic acid
benzyl ester according to methods A-G; off-white solid; IR (solid)
1664, 1519 cm-.sup.1; .sup.1H NMR (400 MHz, d6-DMSO) .delta.
0.78-0.87 (3H, m), 1.18-1.23 (3H, m), 1.99-2.14 (2H, m), 2.55-2.80
(2H, m), 3.19-3.25 (2H, m), 4.54-4.66 (1H, m), 5.20-5.30 (2H, m),
5.35-5.45 (1H, m), 7.47 (1H, m), 7.55-7.71 (1H, m), 8.01 (1H, s),
9.05 (1H, m), 9.31 (1H, s); .sup.19F NMR (376 MHz, d6-DMSO,
proton-decoupled) .delta. -63.11, -139.6, -157.1, -157.2; M+H
634.1, M-H 632.1.
EXAMPLE 23
(S,S)-3-[3-Methyl-2-(2-oxo-3-phenylacetylamino-2H-pyridin-1-yl)-butyrylami-
no]-4-oxo-5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0251] 54
[0252] Prepared from (2-oxo-1,2-dihydro-pyridin-3-yl)-carbamic acid
benzyl ester according to methods A-G; pink solid; IR (solid) 1644,
1683, 1740, 1791 cm-1; .sup.1H NMR (400 MHz, d6-DMSO) .delta. 0.6
(3H, m), 1.0 (3H, m), 2.2-2.3 (1H, m), 2.5-3.0 (2H, m), 3.7-3.8
(2H, m), 4.1-5.4 (4H, m), 6.2-6.3 (1H, m), 7.2-7.4 (5H, m), 7.5-7.7
(2H, m), 8.1-8.2 (1H, m), 8.7-9.2 (2H, m); .sup.19F NMR (376 MHz,
d6-DMSO, proton-decoupled) .delta. -140.6, -141.0, -156.8, -157.0,
-157.2; M+H 606.3, M-H 604.3.
EXAMPLE 24
(S,S)-3-[2-(3-Ethanesulfonylamino-2-oxo-2H-pyridin-1-yl)-3-methyl-butyryla-
mino]-4-oxo-5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0253] 55
[0254] Prepared from (2-oxo-1,2-dihydro-pyridin-3-yl)-carbamic acid
benzyl ester according to methods A-G; off-white solid; IR (solid)
1595, 1646, 1682, 1,742, 1789 cm-1; .sup.1H NMR (400 MHz, d6-DMSO)
.delta. 0.7 (3H, m), 0.9-1.0 (3H, m), 1.2 (3H, m), 2.3 (1H, m),
2.6-3.0 (2H, m), 3.1 (2H, m), 4.1-5.4 (4H, m), 6.3 (1H, m), 7.3
(1H, m), 7.5-7.7 (2H, m), 8.7-9.2 (2H, m); .sup.19F NMR (376 MHz,
d6-DMSO, proton-decoupled) .delta. -140.6, -141.0, -156.7, -157.0,
-157.1; M+H 580.2, M-H 578.3.
EXAMPLE 25
(S)-5-Fluoro-4-oxo-3-[2-(2-oxo-3-propionylamino-2H-pyridin-1-yl)-propionyl-
amino]-pentanoic acid
[0255] 56
[0256] Prepared from (2-oxo-1,2-dihydro-pyridin-3-yl)-carbamic acid
benzyl ester and 3-Amino-5-fluoro-4-hydroxy-pentanoic acid
tert-butyl ester according to methods A-G; pink solid; IR (solid)
1643, 1658, 1711, 1740 cm-1; .sup.1H NMR (400 MHz, d6-DMSO) .delta.
1.0-1.2 (3H, m), 1.4-1.6 (3H, m), 2.4-3.2 (4H, m), 4.2-4.6 (1.5H,
m), 5.0-5.6 (2.5H, m), 6.3 (1H, m), 7.3 (1H, m), 8.2 (1H, m),
8.3-8.8( 1H, m), 9.1 (1H, m); .sup.19F NMR (376 MHz, d6-DMSO,
proton-decoupled) .delta. -226.8, -226.9, -230.6, -231.4, -232.7,
-232.8; M+H 370.4, M-H 368.3.
EXAMPLE 26
(S)-3-[2-(3-Benzoylamino-2-oxo-2H-pyridin-1-yl)-propionylamino]-5-fluoro-4-
-oxo-pentanoic acid
[0257] 57
[0258] Prepared from (2-oxo-1,2-dihydro-pyridin-3-yl)-carbamic acid
benzyl ester and 3-Amino-5-fluoro-4-hydroxy-pentanoic acid
tert-butyl ester according to methods A-G; pink solid; IR (solid)
1523, 1644 cm-1; .sup.1H NMR (400 MHz, d6-DMSO) .delta. 1.6 (3H,
m), 2.5-3.2 (2H, m), 4.2-4.7 (1.5H, m), 5.0-5.6 (2.5H, m), 6.4 (1H,
m), 7.4-7.6 (3H, m), 7.9 (2H, m), 8.3 (1H, m), 8.5-8.9 (1H, m), 9.3
(1H, m); .sup.19F NMR (376 MHz, d6-DMSO, proton-decoupled) .delta.
-226.7, -226.8, -230.4, -231.3, -232.8, -232.9; M+H 418.3, M-H
416.3.
EXAMPLE 27
(S)-3-{2-[3-(2,6-Dichloro-benzoylamino)-2-oxo-2H-pyridin-1-yl]-propionylam-
ino}-5-fluoro-4-oxo-pentanoic acid
[0259] 58
[0260] Prepared from (2-oxo-1,2-dihydro-pyridin-3-yl)-carbamic acid
benzyl ester and 3-Amino-5-fluoro-4-hydroxy-pentanoic acid
tert-butyl ester according to methods A-G; pink solid; IR (solid)
1521, 1646 cm-1; .sup.1H NMR (400 MHz, d6-DMSO) .delta. 1.5-1.6
(3H, m), 2.5-3.2 (2H, m), 4.2-4.7 (1.5H, m), 5.0-5.5 (2.5H, m),
6.3-6.4 (1H, m), 7.4-7.5 (3H, m), 8.3 (1H, m), 8.5-8.9 (1H, m),
10.2 (1H, m); .sup.19F NMR (376 MHz, d6-DMSO, proton-decoupled)
.delta. -226.7, -226.8, -230.6, -231.4, -232.8, -232.9; M+H 486.3,
M-H 484.3.
EXAMPLE 28
(S)-5-Fluoro-4-oxo-3-[2-(2-oxo-3-phenylacetylamino-2H-pyridin-1-yl)-propio-
nylamino]-pentanoic acid
[0261] 59
[0262] Prepared from (2-oxo-1,2-dihydro-pyridin-3-yl)-carbamic acid
benzyl ester and 3-Amino-5-fluoro-4-hydroxy-pentanoic acid
tert-butyl ester according to methods A-G; pink solid; IR (solid)
1524.2, 1652.4 cm-1; .sup.1H NMR (400 MHz, d6-DMSO) .delta. 1.5
(3H, m), 2.5-3.2 (2H, m), 3.8 (2H, m), 4.2-4.7 (1.5H, m), 5.0-5.5
(2.5H, m), 6.3 (1H, m), 7.2-7.4 (6H, m), 8.2 (1H, m), 8.4-8.9 (1H,
m), 9.3 (1H, m); .sup.19F NMR (376 MHz,d6-DMSO, proton-decoupled)
.delta. -226.7, -226.8, -230.6, -231.5, -232.8, -232.9; M+H 432.3,
M-H 430.3.
EXAMPLE 29
(S)-5-Fluoro-4-oxo-3-[2-(2-oxo-3-propionylamino-2H-pyridin-1-yl)-butyrylam-
ino]-pentanoic acid
[0263] 60
[0264] Prepared from (2-oxo-1,2-dihydro-pyridin-3-yl)-carbamic acid
benzyl ester and 3-Amino-5-fluoro-4-hydroxy-pentanoic acid
tert-butyl ester according to methods A-G; pink solid; IR (solid)
1644, 1585, 1518, 1214 cm-1; .sup.1H NMR (400 MHz, d6-DMSO) .delta.
0.8-0.9 (3H, m), 1.05 (3H, t)i 1.9-2.1 (2H, m), 2.4-2.5 (2H, m),
2.6-2.95 (2H, m), 4.2-4.5(2H, m), 5.1-5.5 (3H, m), 6.3-6.35 (1H,
m), 7.4-7.45 (1H, m), 8.2-8.25 (1H, m), 8.8-8.9 (1H, m), 9.1-9.15
(1H, m); .sup.19F NMR (376 MHz, d6-DMSO, proton-decoupled) .delta.
-226.7, -232.6; M+H 384.3, M-H 382.3.
EXAMPLE 30
(S)-3-[2-(3-Benzoylamino-2-oxo-2H-pyridin-1-yl)-butyrylamino]-5-fluoro-4-o-
xo-pentanoic acid
[0265] 61
[0266] Prepared from (2-oxo-1,2-dihydro-pyridin-3-yl)-carbamic acid
benzyl ester and 3-Amino-5-fluoro-4-hydroxy-pentanoic acid
tert-butyl ester according to methods A-G; pink solid; IR (solid)
1643, 1522, 1204 cm-1; .sup.1H NMR (400 MHz, d6-DMSO) .delta.
0.75-0.85 (3H, m), 1.9-2.2 (2H, m), 2.6-2.9 (2H, m), 4.3-4.7(2H,
m), 5.1-5.6 (2H, m6.4-6.5 (1H, m), 7.5-7.85 (4H, m), 7.9-8.0 (1H,
m), 8.3-8.4 (1H, m), 8.85-8.95 (1H, m), 9.35 (1H, s); M+H 432.3,
M-H 430.3.
EXAMPLE 31
(S)-3-{2-[3-(2,6-Dichloro-benzoylamino)-2-oxo-2H-pyridin-1-yl]-butyrylamin-
o}-5-fluoro-4-oxo-pentanoic acid
[0267] 62
[0268] Prepared from (2-oxo-1,2-dihydro-pyridin-3-yl)-carbamic acid
benzyl ester and 3-Amino-5-fluoro-4-hydroxy-pentanoic acid
tert-butyl ester according to methods A-G; white solid; IR (solid)
1682, 1645, 1580, 1516, 1216 cm-1; .sup.1H NMR (400 MHz, d6-DMSO)
.delta. 0.8-0.9 (3H, m), 1.9-2.1 (2H, m), 2.6-2.85 (2H, m),
4.4-4.7(2H, m), 5.1-5.5 (2H, m), 6.4-6.5 (1H, m), 7.5-7.6 (4H, m),
8.33-8.38 (1H, m), 8.85-8.95 (1H, m), 9.15-9.25 (1H, s); M+H 500.3,
M-H 498.3.
EXAMPLE 32
(S)-5-Fluoro-4-oxo-3-(2-{2-oxo-3-[(pyridine-2-carbonyl)-amino]-2H-pyridin--
1-yl)-butyrylamino)-pentanoic acid
[0269] 63
[0270] Prepared from (2-oxo-1,2-dihydro-pyridin-3-yl)-carbamic acid
benzyl ester and 3-Amino-5-fluoro-4-hydroxy-pentanoic acid
tert-butyl ester according to methods A-G; cream solid; IR (solid)
1685, 1644, 1521 cm-1; .sup.1H NMR (400 MHz, d6-DMSO) .delta.
0.81-0.86 (3H, m), 1.90-2.05 (1H, m), 2.06-2.19 (1H, m), 2.54-2.90
(2H, m), 4.58-4.72 (1H, m), 5.07-5.31 (2H, m), 5.42-5.57 (1H, m),
6.40-6.44 (1H, m), 7.47-7.49 (1H, m), 6.68-7.72 (1H, m), 8.09-8.11
(1H, m), 8.18 (1H, d), 8.45-8.47 (1H, m), 8.73-8.75 (1H, m), 8.87
(1H, dd), 10.74 (1H, s), 12.45 (1H, brd s); .sup.19F NMR (376 MHz,
d6-DMSO, proton-decoupled) .delta. -226.8, -230.4, -230.6, -231.0,
-232.5, -232.6, -232.8, -232.9; M+H 433.4, M-H 431.4.
EXAMPLE 33
(S)-5-Fluoro-4-oxo-3-[2-(2-oxo-3-phenylacetylamino-2H-pyridin-1-yl)-butyry-
lamino]-pentanoic acid
[0271] 64
[0272] Prepared from (2-oxo-1,2-dihydro-pyridin-3-yl)-carbamic acid
benzyl ester and 3-Amino-5-fluoro-4-hydroxy-pentanoic acid
tert-butyl ester according to methods A-G; pink solid; IR (solid)
1644, 1672, 1742, 1785 cm-1; 1H NMR (400 MHz, d6-DMSO) .delta.
0.7-0.8 (3H, m), 1.8-2.2 (2H, m), 2.5-3.2 (2H, m), 3.8 (2H, s),
4.2-4.7 (2H, m), 5.1-5.5 (2H, m), 6.3 (1H, m), 7.2-7.4 (6H, m), 8.2
(1H, m), 8.5-9.4 (2H, m); .sup.19F NMR (376 MHz, d6-DMSO,
proton-decoupled) .delta. -226.7, -226.7, -230.4, -231.2, -232.6,
-232.6; M+H 446.3, M-H 444.3.
EXAMPLE 34
(S)-5-Fluoro-4-oxo-3-{2-[2-oxo-3-(2-m-tolyl-acetylamino)-2H-pyridin-1-yl]--
butyrylamino}-pentanoic acid
[0273] 65
[0274] Prepared from (2-oxo-1,2-dihydro-pyridin-3-yl)-carbamic acid
benzyl ester and 3-Amino-5-fluoro-4-hydroxy-pentanoic acid
tert-butyl ester according to methods A-G; ochre solid; IR (solid)
1644, 1678 cm-1; .sup.1H NMR (400 MHz, d6-DMSO) .delta. 0.7-0.8(3H,
m), 1.8-2.2 (2H, m), 2.3 (3H, s), 2.5-3.2 (2H, m), 3.7-3.8 (2H, s),
4.2-5.5 (4H, m), 6.3 (1H, m), 7.0-7.3 (4H, m), 7.4 (1H, m), 8.2
(1H, m), 8.5-8.9 (1H, m), 9.2-9.3 (1H, m); .sup.19F NMR (376 MHz,
d6-DMSO, proton-decoupled) .delta. -226.7, -226.7, -230.4, -231.2,
-232.6, -232.7; M+H 460.3, M-H 459.4.
EXAMPLE 35
(S)-5-Fluoro-4-oxo-3-[2-(2-oxo-3-propionylamino-2H-pyridin-1-yl)-pentanoyl-
amino]-pentanoic acid
[0275] 66
[0276] Prepared from (2-oxo-1,2-dihydro-pyridin-3-yl)-carbamic acid
benzyl ester and 3-Amino-5-fluoro-4-hydroxy-pentanoic acid
tert-butyl ester according to methods A-G; white solid; .sup.1H NMR
(400 MHz, d6-DMSO) .delta. 0 0.85-0.95 (3H, m), 1.0-1.1 (3H, m),
1.1-1.17 (2H, m), 1.9-2.0 (2H, m), 2.4-2.5 (2H, m), 2.6-2.90 (2H,
m), 4.5-4.65 (1H, m), 5.1-5.5 (3H, m), 6.3-6.35 (1H, m), 7.4-7.43
(1H, m), 8.2-8.23 (1H, m), 8.8-8.9 (1H, m), 9.05-9.1 (1H, m);
.sup.9F NMR (376 MHz, d6-DMSO, proton-decoupled) .delta. -226.7,
-232.6; M+H 398.4, M-H 396.4.
EXAMPLE 36
(S)-5-Fluoro-3-[4-methyl-2-(2-oxo-3-propionylamino-2H-pyridin-1-yl)-pentan-
oylamino]-4-oxo-pentanoic acid
[0277] 67
[0278] Prepared from (2-oxo-1,2-dihydro-pyridin-3-yl)-carbamic acid
benzyl ester and 3-Amino-5-fluoro-4-hydroxy-pentanoic acid
tert-butyl ester according to methods A-G; pink solid; .sup.1H NMR
(400 MHz, d6-DMSO) .delta. 0.85(6H, m), 1.05 (3H, t), 1.30 (1H, m),
1.70-2.10 (2H, 2.times.m), 2.30-3.00 (4H, m), 4.60-4.80 (1H, m),
5.05-5.40 (2H, m), 5.65 (1H, m), 6.35 (1H, m), 7.45 (1H, m), 8.25
(1H, m), 8.95 (1H, m), 9.15 (1H, m); .sup.19F NMR (376 MHz,
d6-DMSO, proton-decoupled) .delta. -226.7, -232.5; M+H 412.3.
EXAMPLE 37
(S)-5-Fluoro-3-[2-(5-methyl-2-oxo-3-phenylacetylamino-2H-pyridin-1-yl)-but-
yrylamino]-4-oxo-pentanoic acid
[0279] 68
[0280] Prepared from
(5-Methyl-2-oxo-1,2-dihydro-pyridin-3-yl)-carbamic acid benzyl
ester and 3-Amino-5-fluoro-4-hydroxy-pentanoic acid tert-butyl
ester according to methods A-G; yellow solid; IR (solid) 1654,
1741, 1785 cm-1; .sup.1H NMR (400 MHz, d6-DMSO) .delta. 0.7-0.8
(3H, m), 1.8-2.2 (5H, m), 2.5-3.2 (2H, m), 3.8 (2H, s), 4.2-5.5
(4H, m), 7.1-7.4 (6H, m), 8.1(1H, m), 8.4-8.9 (1H, m), 9.2-9.4 (1H,
m); .sup.19F NMR (376 MHz, d6-DMSO, proton-decoupled) .delta.
-226.7, -226.7, -227.5, -230.5, -231.3, -232.6, -232.6, -233.4; M+H
460.4, M-H 458.4.
EXAMPLE 38
(S,S)-4-Oxo-3-{2-[2-oxo-3-(thiazol-2-ylamino)-2H-pyridin-1-yl]-butyrylamin-
o}-5-(2,3,5,.6-tetrafluoro-phenoxy)-pentanoic acid
[0281] 69
Method I
3-(Thiazol-2-ylamino)-1H-pyridin-2-one
[0282] 70
[0283] To a solution of 3-Amino-1H-pyridin-2-one (2.0 g, 18.7 mmol)
in water (2 mL) was added 15% HCl (10 mL, 18 mmol) followed by
ammonium thiocyanate (1.5 g, 18 mmol) and the mixture was heated to
reflux for two hours. Upon cooling the intermediate thiourea was
found to precipitate as a red-brown solid. The mixture was filtered
and the solid washed with water (5 mL). To a solution of the
thiourea (1.3 g, 7.7 mmol) in EtOH (20 mL) and water (5 mL) was
added chloroacetaldehyde (2.3 mL), 16.4 mmol) and the mixture was
heated to reflux for four hours. On cooling, the mixture was
diluted with EtOAc (30 mL) and washed with 10% NaHCO.sub.3 and
brine. The organic phase was dried over MgSO.sub.4 and concentrated
in vacuo. The residue was purified by flash column chromatography
(100% EtOAc) to afford the title compound as a pale green solid
(1.43 g, 40%): .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 1.6 (1H,
s), 6.45 (1H, t), 6.75 (1H, s), 7.05-7.10 (1H, m), 7.40-7.42 (1H,
m), 8.35-8.5 (2H, m); M+H 194.1, M-H 192.1.
[0284] This intermediate was involved in the sequence described in
methods A and B-G to afford the example 38 as a white solid; IR
(solid) 1648, 1593, 1517, 1490 cm-1; .sup.1H NMR (400 MHz, d6-DMSO)
.delta. 0.75-0.85 (3H, t), 1.9-2.2 (2H, m), 2.6-2.8 (2H, m),
4.6-4.7 (1H, m), 5.2-5.3 (2H, m), 5.35-5.45 (1H, m), 6.3-6.35 (1H,
m), 6.96-6.98 (1H, m), 7.2-7.3 (2H, m), 7.5-7.65 (1H, m), 8.4-8.43
(1H, m), 8.8-8.9(1H, 2.times.d), 9.9 (1H, br s), 12.5 (1H, brd s);
.sup.19F NMR (376 MHz, d6-DMSO, proton-decoupled) .delta. -141.0,
-156.9; M+H 557.2, M-H 555.2.
EXAMPLE 39
(S,S)-4-Oxo-3-[2-(2-oxo-3-propylamino-2H-pyridin-1-yl)-butyrylamino]-5-(2,-
3,5,6-tetrafluoro-phenoxy)-pentanoic acid
[0285] 71
Method J
(S)-2-[3-(Benzyloxycarbonyl-propyl-amino)-2-oxo-2H-pyridin-1-yl]-butyric
acid tert-butyl ester
[0286] 72
[0287] To a solution of
(S)-2-(3-Benzyloxycarbonylamino-2-oxo-2H-pyridin-1- -yl)-butyric
acid tert-butyl ester (100 mg, 0.26 mmol) in anhydrous DMF (3 mL)
was added NaH (60% dispersion, 10 mg, 0.26 mmol) and the reaction
was stirred at ambient temperature for 30 minutes. Propyliodide (30
.mu.L, 0.31 mmol) was added dropwise and the reaction stirred at
ambient temperature overnight. The mixture was concentrated in
vacuo to a solid and partitioned between EtOAc (10 mL) and water
(10 mL). The organic layer was separated, dried over MgSO.sub.4,
and concentrated in vacuo. The residue was purified by flash column
chromatography (30% EtOAc/hexane) to afford the title compound as a
pale green solid (1.43g, 40%): .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 0.85-0.95 (6H, m), 1.35 (9H, s), 1.55-1.65 (2H, m),
1.85-1.95 (1H, m), 2.20-2.27 (1H, m), 3.6-3.7 (2H, m), 5.15-5.2
(2H, m), 5.5-5.6 (1H, m), 6.25 (1H, t), 7.25-7.45 (7H, m); M+H
429.4.
[0288] This intermediate was involved in the sequence described in
methods C-G and finally subjected to hydrogenolysis as described in
method B to afford example 39 as an off-white solid; IR (solid)
1581, 1517, 1489, 938 cm-1; .sup.1H NMR (400 MHz, d6-DMSO) .delta.
0.80 (3H, t), 0.9 (3H, t), 1.5-1.6 (2H, m), 1.8-2.05 (2H, m),
2.5-2.7 (2H, m), 2.9-3.0 (2H, m), 4.6-4.7 (1H, m), 5.1-5.4 (3H, m),
6.1-6.2 (2H, m), 6.85-6.9 (1H, m), 7.5-7.65 (1H, m), 8.7-8.90 (1H,
3.times.d), 12.5 (1H, brd s); M+H 516.2, M-H 514.2.
EXAMPLE 40
[0289] Enzyme Assays
[0290] The assays for caspase inhibition are based on the cleavage
of a fluorogenic substrate by recombinant, purified human
Caspases-1, -3, or -8. The assays are run in essentially the same
way as those reported by Garcia-Calvo et al. (J. Biol. Chem. 273
(1998), 32608-32613), using a substrate specific for each enzyme.
The substrate for Caspase-1 is
Acetyl-Tyr-Val-Ala-Asp-amino-4-methylcoumarin. The substrate for
Caspases -3 and -8 is
Acetyl-Asp-Glu-Val-Asp-amino-4-methylcoumarin. Both substrates are
known in the art.
[0291] The observed rate of enzyme inactivation at a particular
inhibitor concentration, k.sub.obs, is computed by direct fits of
the data to the equation derived by Thornberry et al. (Biochemistry
33 (1994), 3943-3939) using a nonlinear least-squares analysis
computer program (PRISM 2.0; GraphPad software). To obtain the
second order rate constant, k.sub.inact, k.sub.obs values are
plotted against their respective inhibitor concentrations and
k.sub.inact values are subsequently calculated by computerized
linear regression.
[0292] Inhibition of caspases-1,-3, and -8 activity for selected
compounds of this invention was determined by the above method.
Compounds 1-39 inhibited caspase-1 with a k.sub.inact of
>200,000 (M.sup.-1s.sup.-1), caspase-3 with a k.sub.inact of at
>50,000 (k.sub.inact (M.sup.-1s.sup.-1), and caspase-8 with a
k.sub.inact of at >50,000 (k.sub.inact (M.sup.-1s.sup.-1).
EXAMPLE 41
[0293] Inhibition of IL-1.beta. secretion from Whole Blood
[0294] Human blood is freshly drawn from healthy donors and diluted
1:2 in PBS. To 500 .mu.l of diluted blood 50ml of prediluted test
compound in RPMI medium and 10 ml LPS (5 ng/ml final concentration
on the plate) are added (LPS, Serotype 0111:B4, Sigma L3012). After
stimulation for 18 hours supernatants are collected and assayed for
IL-1.beta. levels using the appropriate ELISA kit (R&D
systems).
[0295] Table 2 below shows inhibition of IL-1.beta. secretion from
human whole blood for selected compounds of this invention as
determined individually by the above methods.
2TABLE 2 Inhibition of IL-1.beta. secretion Compound Number
IC.sub.50 (.mu.M) 1, 2, 3, 5, 7, 10, 11, 14, 17, & 29 <0.5
4, 6, 8, 9, 12, 13, 15, 16, 18, 19, 20, 21, 0.5-5 22, 23, 28., 36,
38, & 39
EXAMPLE 42
[0296] Hypoxia-Induced Apoptosis of Rat Cortical Neurons
[0297] Cortical neurons are dissociated from Wistar rat embryos
(E17) by a modification of the procedure of Rogers et al. 1997,
Brain Res. Bulletin, 44:131. Briefly, cerebral cortices are
isolated aseptically from 15-20 Wistar rat embryos. A cell
suspension is prepared by mincing the cerebral cortices and
digesting them with papain. Cells are washed with ovomucoid enzyme
inhibitor and DNaseI and plated onto Poly-D lysine coated plates in
high glucose DMEM containing 10% heat-inactivated fetal calf serum,
L-glutamine, penicillin and streptomycin. The yield of neurons is
10.times.7 per embryo and they are 80-90% viable as assessed by
Trypan blue exclusion.
[0298] The neurons are cultured in complete medium at 37 .degree.
C. in a normal atmosphere for 48 hours prior to the hypoxia
experiments. For hypoxia, the normal cell medium is replaced by
oxygen-depleted serum-free medium. Cells are incubated in an
atmosphere of 95% N2/5% CO.sub.2 for various lengths of time.
Compounds are dissolved in DMSO at 100 mM then diluted in medium
and added to the culture from time=0. The level of apoptosis is
measured using a Cell Death Detection ELISA kit (Roche) which
detects DNA fragmentation. Plates are read at 405 nm. Controls
included cells cultured in aerobic conditions in serum-containing
medium (+serum) and cells cultured in aerobic conditions in
serum-deprived medium (-serum).
[0299] Table 3 shows the results of the activity of selected
compounds of this invention tested individually in the
Hypoxia-induced apoptosis of rat cortical neurons.
3TABLE 3 Activity in Hypoxia-induced Apoptosis Assay Compound
Number IC.sub.50 (.mu.M) 2, 5, 10, 11, 12, 13, 15, 17, 20, 22, 38,
& <1 39 1, 3, 4, 6, 7, 8, 9, 14, 16, 18, 19, 21, 24, 1-10
29, 30, 31, 34, 35, 36 & 37
EXAMPLE 43
[0300] Anti-Fas Induced Apoptosis Assay
[0301] Cellular apoptosis may be induced by the binding of Fas
ligand (FasL) to its receptor, CD95 (Fas). CD95 is one of a family
of related receptors, known as death receptors, which can trigger
apoptosis in cells via activation of the caspase enzyme cascade.
The process is initiated by the binding of the adapter molecule
FADD/MORT-1 to the cytoplasmic domain of the CD-95 receptor-ligand
complex. Caspase-8 then binds FADD and becomes activated,
initiating a cascade of events that involve the activation of
downstream caspases and subsequent cellular apoptosis. Apoptosis
can also be induced in cells expressing CD95 e.g., the Jurkat E6.1
T cell lymphoma cell line, using an antibody, rather than FasL, to
crosslink the cell surface CD95. Anti-Fas-induced apoptosis is also
triggered via the activation of caspase-8. This provides the basis
of a cell based assay to screen compounds for inhibition of the
caspase-8-mediated apoptotic pathway.
[0302] Experimental Procedure
[0303] Jurkat E6.1 cells are cultured in complete medium consisting
of RPMI-1640 (Sigma No)+10% foetal calf serum (Gibco BRL
No.10099-141)+2mM L-glutamine (Sigma No. G-7513). The cells are
harvested in log phase of growth. 100 ml of cells at 5-8.times.105
cells/ml are transferred to sterile 50 ml Falcon centrifuge tubes
and centrifuged for 5 minutes at 100.times.g at room temperature.
The supernatant is removed and the combined cell pellets
resuspended in 25 ml of complete medium. The cells are counted and
the density adjusted to 2.times.106 cells/ml with complete
medium.
[0304] The test compound is dissolved in dimethyl sulfoxide
(DMSO)(Sigma No. D-2650) to give a 100 mM stock solution. This is
diluted to 400 .mu.M in complete medium, then serially diluted in a
96-well plate prior to addition to the cell assay plate.
[0305] 100 .mu.l of the cell suspension (2.times.106 cells) is
added to each well of a sterile 96-well round-bottomed cluster
plate (Costar No. 3790). 50 .mu.l of compound solution at the
appropriate dilution and 50 .mu.l of anti-Fas antibody, clone CH-11
(Upstate, Cat No.1 544 675) at a final concentration of 10 ng/ml,
are added to the wells. Control wells are set up minus antibody and
minus compound but with a serial dilution of DMSO as vehicle
control. The plates are incubated for 16-18 hrs at 37.degree. C. in
5% CO.sub.2 and 95% humidity.
[0306] Apoptosis of the cells is measured by the quantitation of
DNA fragmentation using a `Cell Death Detection Assay` from Roche
diagnostics, No. 1544 675. After incubation for 16-18 hrs the assay
plates are centrifuged at 100.times.g at room temperature for 5
minutes. 150 .mu.l of the supernatant are removed and replaced by
150 .mu.l of fresh complete medium. The cells are then harvested
and 200 .mu.l of the lysis buffer supplied in the assay kit are
added to each well. The cells are triturated to ensure complete
lysis and incubated for 30 minutes at 4.degree. C. The plates are
then centrifuged at 1900.times.g for 10 minutes and the
supernatants diluted 1:20 in the incubation buffer provided. 100
.mu.l of this solution is then assayed according to the
manufacturer's instructions supplied with the kit. OD405 nm is
measured 20 minutes after addition of the final substrate in a
SPECTRAmax Plus plate reader (Molecular Devices). OD405 nm is
plotted versus compound concentration and the IC50 values for the
compounds are calculated using the curve-fitting program SOFTmax
Pro (Molecular Devices) using the four parameter fit option.
[0307] Selected compounds have been tested in this assay and shown
to inhibit Fas-induced apoptosis of Jurkat cells with IC50 values
between 0.001 .mu.M and 0.15 .mu.M.
4TABLE 4 Activity in FAS-induced Apoptosis Assay Compound Number
IC.sub.50 (.mu.M) 1, 2, 4, 5, 7, 11, 13, 17, 18, 19, 22, 25,
<0.5 27, 29, 30, 31, 32, 33, 34, 35, 37 0.5-2 26, 28, 36
[0308] While we have described a number of embodiments of this
invention, it is apparent that our basic examples may be altered to
provide other embodiments which utilize the compounds and methods
of this invention. Therefore, it will be appreciated that the scope
of this invention is to be defined by the appended claims rather
than by the specific embodiments that have been represented by way
of example above.
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