U.S. patent application number 12/630350 was filed with the patent office on 2010-04-29 for caspase inhibitors and uses thereof.
This patent application is currently assigned to VERTEX PHARMACEUTICALS INCORPORATED. Invention is credited to Jean-Damien Charrier, Steven Durrant, Adam Looker, Michael Mortimore, Michael O'Donnell, Sharn Ramaya, Alistair Rutherford, John R. Studley, Martin Trudeau.
Application Number | 20100105914 12/630350 |
Document ID | / |
Family ID | 34923260 |
Filed Date | 2010-04-29 |
United States Patent
Application |
20100105914 |
Kind Code |
A1 |
Charrier; Jean-Damien ; et
al. |
April 29, 2010 |
CASPASE INHIBITORS AND USES THEREOF
Abstract
The present invention provides a compound of formula I:
##STR00001## wherein the variables are as defined herein. The
present invention also provides processes for preparing the
compounds of formula I, and intermediates thereof, pharmaceutical
compositions comprising those compounds, and methods of using the
compounds and compositions.
Inventors: |
Charrier; Jean-Damien;
(Grove, GB) ; Durrant; Steven; (Abingdon, GB)
; Mortimore; Michael; (Burford, GB) ; O'Donnell;
Michael; (Abingdon, GB) ; Rutherford; Alistair;
(Abingdon, GB) ; Ramaya; Sharn; (Burghfield
Common, GB) ; Studley; John R.; (Abingdon, GB)
; Trudeau; Martin; (Shannon, CA) ; Looker;
Adam; (Cambridge, MA) |
Correspondence
Address: |
VERTEX PHARMACEUTICALS INC.
130 WAVERLY STREET
CAMBRIDGE
MA
02139-4242
US
|
Assignee: |
VERTEX PHARMACEUTICALS
INCORPORATED
CAMBRIDGE
MA
|
Family ID: |
34923260 |
Appl. No.: |
12/630350 |
Filed: |
December 3, 2009 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11069895 |
Feb 28, 2005 |
7652153 |
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12630350 |
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60548610 |
Feb 27, 2004 |
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60629743 |
Nov 19, 2004 |
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60629661 |
Nov 19, 2004 |
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Current U.S.
Class: |
546/112 ;
546/279.1; 548/195; 548/517 |
Current CPC
Class: |
A61P 5/14 20180101; A61P
31/04 20180101; A61P 11/06 20180101; A61P 31/06 20180101; C07D
401/14 20130101; A61P 27/02 20180101; A61P 7/04 20180101; A61P
17/06 20180101; A61P 43/00 20180101; A61P 31/12 20180101; Y02A
50/30 20180101; A61P 11/16 20180101; A61P 31/14 20180101; C07D
209/52 20130101; A61P 19/00 20180101; A61P 7/06 20180101; A61P
31/18 20180101; A61P 35/00 20180101; A61P 17/02 20180101; A61P 1/16
20180101; A61P 9/04 20180101; A61P 17/00 20180101; A61P 25/28
20180101; A61K 38/06 20130101; A61P 3/10 20180101; A61P 7/00
20180101; A61P 11/00 20180101; A61P 25/08 20180101; A61P 1/18
20180101; A61P 37/08 20180101; A61P 37/06 20180101; C07K 5/0202
20130101; A61P 35/02 20180101; A61P 37/00 20180101; A61P 19/08
20180101; Y02A 50/389 20180101; A61P 17/16 20180101; A61P 19/10
20180101; C07D 207/16 20130101; A61P 1/04 20180101; A61P 9/10
20180101; A61P 13/12 20180101; A61P 21/00 20180101; A61P 21/02
20180101; Y02A 50/385 20180101; A61P 37/02 20180101; C07D 401/12
20130101; A61P 9/00 20180101; A61P 25/00 20180101; A61P 25/16
20180101; A61P 9/08 20180101; A61P 25/14 20180101; C07D 405/12
20130101; A61P 25/32 20180101; A61P 19/02 20180101; C07D 417/06
20130101; A61P 1/00 20180101; A61P 31/20 20180101; A61P 17/14
20180101; A61P 21/04 20180101; C07D 417/12 20130101; A61P 17/04
20180101; A61P 31/00 20180101; Y02A 50/387 20180101; A61P 29/00
20180101 |
Class at
Publication: |
546/112 ;
546/279.1; 548/195; 548/517 |
International
Class: |
C07D 405/14 20060101
C07D405/14; C07D 417/14 20060101 C07D417/14; C07D 405/12 20060101
C07D405/12 |
Claims
1-36. (canceled)
37. A process for preparing a compound of formula I: ##STR00382##
wherein Y is: ##STR00383## R.sup.1 is H, C.sub.1-12aliphatic,
C.sub.3-10cycloaliphatic, C.sub.6-10aryl, 5-10 membered
heterocyclyl, 5-10 membered heteroaryl,
(C.sub.3-10cycloalkyl)-(C.sub.1-12aliphatic)-,
cycloalkenyl-(C.sub.1-12aliphatic)-,
(C.sub.6-10aryl)-(C.sub.1-12aliphatic)-, (5-10 membered
heterocyclyl)-(C.sub.1-12aliphatic)-, or (5-10 membered
heteroaryl)-(C.sub.1-12aliphatic)-, wherein any hydrogen atom is
optionally and independently replaced by R.sup.8 and any set of two
hydrogen atoms bound to the same atom is optionally and
independently replaced by carbonyl; Ring A is: ##STR00384##
wherein, in each ring, any hydrogen atom is optionally and
independently replaced by R.sup.4 and any set of two hydrogen atoms
bound to the same atom is optionally and independently replaced by
carbonyl; when Ring A is ##STR00385## then R is R.sup.3C(O)--,
HC(O), R.sup.3SO.sub.2--, R.sup.3OC(O), (R.sup.3).sub.2NC(O),
(R.sup.3)(H)NC(O), R.sup.3C(O)C(O)--, R.sup.3--,
(R.sup.3).sub.2NC(O)C(O), (R.sup.3)(H)NC(O)C(O), or
R.sup.3OC(O)C(O)--; and R.sup.3 is C.sub.1-12aliphatic,
C.sub.3-10cycloaliphatic, C.sub.6-10aryl, 5-10 membered
heterocyclyl, 5-10 membered heteroaryl,
(C.sub.3-10cycloaliphatic)-(C.sub.1-12aliphatic)-,
(C.sub.6-10aryl)-(C.sub.1-12aliphatic)-, (5-10 membered
heterocyclyl)-(C.sub.1-12aliphatic)-, or (5-10 membered
heteroaryl)-(C.sub.1-12aliphatic)-; or two R.sup.3 groups bound to
the same atom form together with that atom a 3-10 membered aromatic
or nonaromatic ring; wherein any ring is optionally fused to an
C.sub.6-10aryl, 5-10 membered heteroaryl, C.sub.3-10cycloalkyl, or
5-10 membered heterocyclyl; wherein up to 3 aliphatic carbon atoms
may be replaced by a group selected from O, N, NR.sup.9, S, SO, and
SO.sub.2, wherein R.sup.3 is substituted with up to 6 substituents
independently selected from R.sup.8'; when Ring A is ##STR00386##
then R is R.sup.3C(O)--, as shown in formula II, ##STR00387## and
R.sup.3 is phenyl, thiophene, or pyridine, wherein each ring is
optionally substituted with up to 5 groups independently selected
from R.sup.8', and wherein at least one position on the phenyl,
thiophene, or pyridine adjacent to bond x is substituted by
R.sup.12, wherein R.sup.12 has no more than 5 straight-chained
atoms; R.sup.4 is halogen, --OR.sup.9, --NO.sub.2--CN --CF.sub.3,
--OCF.sub.1, --R.sup.9, 1,2-methylenedioxy, 1,2-ethylenedioxy,
--N(R.sup.9).sub.2, --SR.sup.9, --SOR.sup.9,
--SO.sub.2R.sup.9--SO.sub.2N(R.sup.9).sub.2, --SO.sub.3R.sup.9,
--C(O)R.sup.9, --C(O)C(O)R.sup.9, --C(O)C(O)OR.sup.9,
--C(O)C(O)N(R.sup.9).sub.2, --C(O)CH.sub.2C(O)R.sup.9,
--C(S)R.sup.9, --C(S)OR.sup.9, --C(O)OR.sup.9, --OC(O)R.sup.9,
--C(O)N(R.sup.9).sub.2, --OC(O)N(R.sup.9).sub.2,
--C(S)N(R.sup.9).sub.2, --(CH.sub.2).sub.0-2NHC(O)R.sup.9,
--N(R.sup.9)N(R.sup.9)COR.sup.9,
--N(R.sup.9)N(R.sup.9)C(O)OR.sup.9,
--N(R.sup.9)N(R.sup.9)CON(R.sup.9).sub.2,
--N(R.sup.9)SO.sub.2R.sup.9, --N(R.sup.9)SO.sub.2N(R.sup.9).sub.2,
--N(R.sup.9)C(O)OR.sup.9, --N(R.sup.9)C(O)R.sup.9,
--N(R.sup.9)C(S)R.sup.9, --N(R.sup.9)C(O)N(R.sup.9).sub.2,
--N(R.sup.9)C(S)N(R.sup.9).sub.2--N(COR.sup.9)COR.sup.9,
--N(OR.sup.9)R.sup.9, --C(.dbd.NH)N(R.sup.9).sub.2,
--C(O)N(OR.sup.9)R.sup.9, --C(.dbd.NOR.sup.9)R.sup.9,
--OP(O)(OR.sup.9).sub.2, --P(O)(R.sup.9).sub.2,
--P(O)(OR.sup.9).sub.2, or --P(O)(H)(OR.sup.9); R.sup.2 is
--C(R.sup.5)(R.sup.6)(R.sup.7), C.sub.6-10aryl, 5-10 membered
heteroaryl, or C.sub.3-7 cycloalkyl; R.sup.5 is H or a C.sub.1-6
straight-chained or branched alkyl; R.sup.6 is H or a C.sub.1-6
straight-chained or branched alkyl; R.sup.7 is --CF.sub.3,
--C.sub.3-7cycloalkyl, C.sub.6-10aryl, 5-10 membered heteroaryl,
heterocycle, or a C.sub.1-6 straight-chained or branched alkyl,
wherein each carbon atom of the alkyl is optionally and
independently substituted with R.sup.10; or R.sup.5 and R.sup.7
taken together with the carbon atom to which they are attached form
a 3-10 membered cycloaliphatic; R.sup.8 and R.sup.8' are each
independently halogen, --OR.sup.9, --NO.sub.2, --CN, --CF.sub.3,
--OCF.sub.3, --R.sup.9, 1,2-methylenedioxy, 1,2-ethylenedioxy,
--N(R.sup.9).sub.2, --SR.sup.9, --SOR.sup.9, --SO.sub.2R.sup.9,
--SO.sub.2N(R.sup.9).sub.2--SO.sub.3R.sup.9, --C(O)R.sup.9,
--C(O)C(O)R.sup.9, --C(O)C(O)OR.sup.9, --C(O)C(O)N(R.sup.9).sub.2,
--C(O)CH.sub.2C(O)R.sup.9, --C(S)R.sup.9, --C(S)OR.sup.9,
--C(O)OR.sup.9, --OC(O)R.sup.9, --C(O)N(R.sup.9).sub.2,
--OC(O)N(R.sup.9).sub.2, --C(S)N(R.sup.9).sub.2,
--(CH.sub.2).sub.0-2NHC(O)R.sup.9, --N(R.sup.9)N(R.sup.9)COR.sup.9,
--N(R.sup.9)N(R.sup.9)C(O)OR.sup.9,
--N(R.sup.9)N(R.sup.9)CON(R.sup.9).sub.2,
--N(R.sup.9)SO.sub.2R.sup.9, --N(R.sup.9)SO.sub.2N(R.sup.9).sub.2,
--N(R.sup.9)C(O)OR.sup.9, --N(R.sup.9)C(O)R.sup.9,
--N(R.sup.9)C(S)R.sup.9, --N(R.sup.9)C(O)N(R.sup.9).sub.2,
--N(R.sup.9)C(S)N(R.sup.9).sub.2, --N(COR.sup.9)COR.sup.9,
--N(OR.sup.9)R.sup.9, --C(.dbd.NH)N(R.sup.9).sub.2,
--C(O)N(OR.sup.9)R.sup.9, --C(.dbd.NOR.sup.9)R.sup.9,
--OP(O)(OR.sup.9).sub.2, --P(O)(R.sup.9).sub.2,
--P(O)(OR.sup.9).sub.2, and --P(O)(H)(OR.sup.9); R.sup.9 is
hydrogen, C.sub.1-12aliphatic, C.sub.3-10cycloaliphatic,
C.sub.6-10aryl, 5-10 membered heterocyclyl, 5-10 membered
heteroaryl, (C.sub.3-10cycloaliphatic)-(C.sub.1-12aliphatic)-,
(C.sub.6-10aryl)-(C.sub.1-12aliphatic)-, (5-10 membered
heterocyclyl)-(C.sub.1-12aliphatic)-, or
heteroaryl-(C.sub.1-12aliphatic)-; wherein any hydrogen atom is
optionally and independently replaced by R.sup.13 and any set of
two hydrogen atoms bound to the same atom is optionally and
independently replaced by carbonyl; R.sup.10 is halogen,
--OR.sup.11, --NO.sub.2, --CN, --CF.sub.3--OCF.sub.3, --R.sup.11,
or --SR.sup.11; wherein R.sup.11 is C.sub.1-4-aliphatic-; R.sup.11
is C.sub.1-4-aliphatic-; R.sup.12 is halogen, --OR.sup.11,
--NO.sub.2--CN --CF.sub.3--OCF.sub.3, --R.sup.11, or --SR.sup.9;
R.sup.13 is --OR.sup.14, --NO.sub.2, --CN, --CF.sub.3, --OCF.sub.3,
--R.sup.14, 1,2-methylenedioxy, 1,2-ethylenedioxy,
--N(R.sup.14).sub.2, --SR.sup.14, --SOR.sup.14,
--SO.sub.7R.sup.14--SO.sub.2N(R.sup.14).sub.2--SO.sub.3R.sup.14,
--C(O)R.sup.14, --C(O)C(O)R.sup.14, --C(O)C(O)OR.sup.14,
--C(O)C(O)N(R.sup.14).sub.2,
--C(O)CH.sub.2C(O)R.sup.14--C(S)R.sup.14, --C(S)OR.sup.14,
--C(O)OR.sup.14, --OC(O)R.sup.14, --C(O)N(R.sup.14).sub.2,
--OC(O)N(R.sup.14).sub.2, --C(S)N(R.sup.14).sub.2,
--(CH.sub.2).sub.0-2NHC(O)R.sup.14,
--N(R.sup.14)N(R.sup.14)COR.sup.14,
--N(R.sup.14)N(R.sup.14)C(O)OR.sup.14,
--N(R.sup.14)N(R.sup.14)CON(R.sup.14).sub.2,
--N(R.sup.14)SO.sub.2R.sup.14,
--N(R.sup.14)SO.sub.2N(R.sup.14).sub.2, --N(R.sup.14)C(O)OR.sup.14,
--N(R.sup.14)C(O)R.sup.14, --N(R.sup.14)C(S)R.sup.14,
--N(R.sup.14)C(O)N(R.sup.14).sub.2,
--N(R.sup.14)C(S)N(R.sup.14).sub.2, --N(COR.sup.14)COR.sup.14,
--N(OR.sup.14)R.sup.14, --C(.dbd.NH)N(R.sup.14).sub.2,
--C(O)N(OR.sup.14)R.sup.14, --C(.dbd.NOR.sup.14)R.sup.14,
--OP(O)(OR.sup.14).sub.2, --P(O)(R.sup.14).sub.2,
--P(O)(OR.sup.14).sub.2, and --P(O)(H)(OR.sup.14); R.sup.14 is
hydrogen, C.sub.1-12aliphatic, C.sub.3-10cycloaliphatic,
C.sub.6-10aryl, 5-10 membered heterocyclyl, 5-10 membered
heteroaryl, (C.sub.3-10cycloaliphatic)-(C.sub.1-12aliphatic),
(C.sub.6-10aryl)-(C.sub.1-12aliphatic)-, (5-10 membered
heterocyclyl)-(C.sub.1-12aliphatic)-, or
heteroaryl-(C.sub.1-12aliphatic)-; comprising reacting a compound
of formula 1: ##STR00388## wherein R.sup.1, R.sup.2, and Ring A are
as defined above; and a compound of formula RX, wherein R is as
defined above and --X is OH or an appropriate derivative or leaving
group, in the presence of conditions for coupling an amine and an
acid (when X is OH) or appropriate acid derivative (when X is an
appropriate leaving group) to provide the compound of formula
I.
38. A process for preparing a compound of formula I: ##STR00389##
wherein Y is: ##STR00390## R.sup.1 is H, C.sub.1-12aliphatic,
C.sub.3-10cycloaliphatic, C.sub.6-10aryl, 5-10 membered
heterocyclyl, 5-10 membered heteroaryl,
(C.sub.3-10cycloalkyl)-(C.sub.1-12aliphatic)-,
cycloalkenyl-(C.sub.1-12aliphatic)-,
(C.sub.6-10aryl)-(C.sub.1-12aliphatic)-, (5-10 membered
heterocyclyl)-(C.sub.1-12aliphatic)-, or (5-10 membered
heteroaryl)-(C.sub.1-12aliphatic)-, wherein any hydrogen atom is
optionally and independently replaced by R.sup.8 and any set of two
hydrogen atoms bound to the same atom is optionally and
independently replaced by carbonyl; Ring A is: ##STR00391##
wherein, in each ring, any hydrogen atom is optionally and
independently replaced by R.sup.4 and any set of two hydrogen atoms
bound to the same atom is optionally and independently replaced by
carbonyl; when Ring A is ##STR00392## then R is R.sup.3C(O)--,
HC(O), R.sup.3SO.sub.2--, R.sup.3OC(O), (R.sup.3).sub.2NC(O),
(R.sup.3)(H)NC(O), R.sup.3C(O)C(O)--, R.sup.3--,
(R.sup.3).sub.2NC(O)C(O), (R.sup.3)(H)NC(O)C(O), or
R.sup.3OC(O)C(O)--; and R.sup.3 is C.sub.1-12aliphatic,
C.sub.3-10-cycloaliphatic, C.sub.6-10aryl, 5-10 membered
heterocyclyl, 5-10 membered heteroaryl,
(C.sub.3-10cycloaliphatic)-(C.sub.1-12aliphatic)-,
(C.sub.6-10aryl)-(C.sub.1-12aliphatic)-, (5-10 membered
heterocyclyl)-(C.sub.1-12aliphatic)-, or (5-10 membered
heteroaryl)-(C.sub.1-12aliphatic)-; or two R.sup.3 groups bound to
the same atom form together with that atom a 3-10 membered aromatic
or nonaromatic ring; wherein any ring is optionally fused to an
C.sub.6-10aryl, 5-10 membered heteroaryl, C.sub.3-10cycloalkyl, or
5-10 membered heterocyclyl; wherein up to 3 aliphatic carbon atoms
may be replaced by a group selected from O, N, NR.sup.9, S, SO, and
SO.sub.2, wherein R.sup.3 is substituted with up to 6 substituents
independently selected from R.sup.8'; when Ring A is ##STR00393##
then R is R.sup.3C(O)--, as shown in formula II, ##STR00394## and
R.sup.3 is phenyl, thiophene, or pyridine, wherein each ring is
optionally substituted with up to 5 groups independently selected
from R.sup.8', and wherein at least one position on the phenyl,
thiophene, or pyridine adjacent to bond x is substituted by
R.sup.12, wherein R.sup.12 has no more than 5 straight-chained
atoms; R.sup.4 is halogen, --OR.sup.9, --NO.sub.2, --CN,
--CF.sub.3, --OCF.sub.3, --R.sup.9, 1,2-methylenedioxy,
1,2-ethylenedioxy, --N(R.sup.9).sub.2, --SR.sup.9, --SOR.sup.9,
--SO.sub.2R.sup.9--SO.sub.2N(R.sup.9).sub.2, --SO.sub.3R.sup.9,
--C(O)R.sup.9, --C(O)C(O)R.sup.9, --C(O)C(O)OR.sup.9,
--C(O)C(O)N(R.sup.9).sub.2, --C(O)CH.sub.2C(O)R.sup.9,
--C(S)R.sup.9, --C(S)OR.sup.9, --C(O)OR.sup.9, --OC(O)R.sup.9,
--C(O)N(R.sup.9).sub.2, --OC(O)N(R.sup.9).sub.2,
--C(S)N(R.sup.9).sub.2, --(CH.sub.2).sub.0-2NHC(O)R.sup.9,
--N(R.sup.9)N(R.sup.9)COR.sup.9,
--N(R.sup.9)N(R.sup.9)C(O)OR.sup.9,
--N(R.sup.9)N(R.sup.9)CON(R.sup.9).sub.2,
--N(R.sup.9)SO.sub.2R.sup.9, --N(R.sup.9)SO.sub.2N(R.sup.9).sub.2,
--N(R.sup.9)C(O)OR.sup.9, --N(R.sup.9)C(O)R.sup.9,
--N(R.sup.9)C(S)R.sup.9, --N(R.sup.9)C(O)N(R.sup.9).sub.2,
--N(R.sup.9)C(S)N(R.sup.9).sub.2, --N(COR.sup.9)COR.sup.9,
--N(OR.sup.9)R.sup.9, --C(.dbd.NH)N(R.sup.9).sub.2,
--C(O)N(OR.sup.9)R.sup.9, --C(.dbd.NOR.sup.9)R.sup.9,
--OP(O)(OR.sup.9).sub.2, --P(O)(R.sup.9).sub.2,
--P(O)(OR.sup.9).sub.2, or --P(O)(H)(OR.sup.9); R.sup.2 is
--C(R.sup.5)(R.sup.6)(R.sup.7), C.sub.6-10aryl, 5-10 membered
heteroaryl, or C.sub.3-7 cycloalkyl; R.sup.5 is H or a C.sub.1-6
straight-chained or branched alkyl; R.sup.6 is H or a C.sub.1-6
straight-chained or branched alkyl; R.sup.7 is --CF.sub.3,
--C.sub.3-7cycloalkyl, C.sub.6-10aryl, 5-10 membered heteroaryl,
heterocycle, or a C.sub.1-6 straight-chained or branched alkyl,
wherein each carbon atom of the alkyl is optionally and
independently substituted with R.sup.10; or R.sup.5 and R.sup.7
taken together with the carbon atom to which they are attached form
a 3-10 membered cycloaliphatic; R.sup.8 and R.sup.8' are each
independently halogen, --OR.sup.9, --NO.sub.2, --CN, --CF.sub.3,
--OCF.sub.3, --R.sup.9, 1,2-methylenedioxy, 1,2-ethylenedioxy,
--N(R.sup.9).sub.2, --SR.sup.9, --SOR.sup.9, --SO.sub.2R.sup.9,
--SO.sub.2N(R.sup.9).sub.2, --SO.sub.3R.sup.9, --C(O)R.sup.9,
--C(O)C(O)R.sup.9, --C(O)C(O)OR.sup.9, --C(O)C(O)N(R.sup.9).sub.2,
--C(O)CH.sub.2C(O)R.sup.9, --C(S)R.sup.9, --C(S)OR.sup.9,
--C(O)OR.sup.9, --OC(O)R.sup.9, --C(O)N(R.sup.9).sub.2,
--OC(O)N(R.sup.9).sub.2, --C(S)N(R.sup.9).sub.2,
--(CH.sub.2).sub.0-2NHC(O)R.sup.9, --N(R.sup.9)N(R.sup.9)COR.sup.9,
--N(R.sup.9)N(R.sup.9)C(O)OR.sup.9,
--N(R.sup.9)N(R.sup.9)CON(R.sup.9).sub.2,
--N(R.sup.9)SO.sub.2R.sup.9, --N(R.sup.9)SO.sub.2N(R.sup.9).sub.2,
--N(R.sup.9)C(O)OR.sup.9, --N(R.sup.9)C(O)R.sup.9,
--N(R.sup.9)C(S)R.sup.9, --N(R.sup.9)C(O)N(R.sup.9).sub.2,
--N(R.sup.9)C(S)N(R.sup.9).sub.2, --N(COR.sup.9)COR.sup.9,
--N(OR.sup.9)R.sup.9, --C(.dbd.NH)N(R.sup.9).sub.2,
--C(O)N(OR.sup.9)R.sup.9, --C(.dbd.NOR.sup.9)R.sup.9,
--OP(O)(OR.sup.9).sub.2, --P(O)(R.sup.9).sub.2,
--P(O)(OR.sup.9).sub.2 and --P(O)(H)(OR.sup.9); R.sup.9 is
hydrogen, C.sub.1-12aliphatic, C.sub.3-10cycloaliphatic,
C.sub.6-10aryl, 5-10 membered heterocyclyl, 5-10 membered
heteroaryl, (C.sub.3-10cycloaliphatic)-(C.sub.1-12aliphatic)-,
(C.sub.6-10aryl)-(C.sub.1-12aliphatic)-, (5-10 membered
heterocyclyl)-(C.sub.1-12aliphatic)-, or
heteroaryl-(C.sub.1-12aliphatic)-; wherein any hydrogen atom is
optionally and independently replaced by R.sup.13 and any set of
two hydrogen atoms bound to the same atom is optionally and
independently replaced by carbonyl; R.sup.12 is halogen,
--OR.sup.11, --NO.sub.2, --CN, --CF.sub.3--OCF.sub.3, --R.sup.11,
or --SR.sup.11; wherein R.sup.11 is C.sub.1-4-aliphatic-; R.sup.11
is C.sub.1-4-aliphatic-; R.sup.12 is halogen, --OR.sup.11,
--NO.sub.2, --CN, --CF.sub.3, --OCF.sub.3, --R.sup.11, or
--SR.sup.9; R.sup.13 is --OR.sup.14, --NO.sub.2, --CN, --CF.sub.3,
--OCF.sub.3, --R.sup.14, 1,2-methylenedioxy, 1,2-ethylenedioxy,
--N(R.sup.14).sub.2, --SR.sup.14, --SOR.sup.14,
--SO.sub.2R.sup.14--SO.sub.2N(R.sup.14).sub.2, --SO.sub.3R.sup.14,
--C(O)R.sup.14, --C(O)C(O)R.sup.14, --C(O)C(O)OR.sup.14,
--C(O)C(O)N(R.sup.14).sub.2,
--C(O)CH.sub.2C(O)R.sup.14--C(S)R.sup.14, --C(S)OR.sup.14,
--C(O)OR.sup.14, --OC(O)R.sup.14, --C(O)N(R.sup.14).sub.2,
--OC(O)N(R.sup.14).sub.2, --C(S)N(R.sup.14).sub.2,
--(CH.sub.2).sub.0-2NHC(O)R.sup.14,
--N(R.sup.14)N(R.sup.14)COR.sup.14,
--N(R.sup.14)N(R.sup.14)C(O)OR.sup.14,
--N(R.sup.14)N(R.sup.14)CON(R.sup.14).sub.2,
--N(R.sup.14)SO.sub.2R.sup.14,
--N(R.sup.14)SO.sub.2N(R.sup.14).sub.2, --N(R.sup.14)C(O)OR.sup.14,
--N(R.sup.14)C(O)R.sup.14, --N(R.sup.14)C(S)R.sup.14,
--N(R.sup.14)C(O)N(R.sup.14).sub.2,
--N(R.sup.14)C(S)N(R.sup.14).sub.2, --N(COR.sup.14)COR.sup.14,
--N(OR.sup.14)R.sup.14, --C(.dbd.NH)N(R.sup.14).sub.2,
--C(O)N(OR.sup.14)R.sup.14, --C(.dbd.NOR.sup.14)R.sup.14,
--OP(O)(OR.sup.14).sub.2, --P(O)(R.sup.14).sub.2,
--P(O)(OR.sup.14).sub.2, and --P(O)(H)(OR.sup.14); R.sup.14 is
hydrogen, C.sub.1-12aliphatic, C.sub.3-10cycloaliphatic,
C.sub.6-10aryl, 5-10 membered heterocyclyl, 5-10 membered
heteroaryl, (C.sub.3-10cycloaliphatic)-(C.sub.1-12aliphatic)-,
(C.sub.6-10aryl)-(C.sub.1-12aliphatic)-, (5-10 membered
heterocyclyl)-(C.sub.1-12aliphatic)-, or
heteroaryl-(C.sub.1-12aliphatic)-; comprising reacting a compound
of formula 7-A: ##STR00395## wherein Ring A and R.sup.1 are as
defined above; and a compound of formula RNHCH(R.sup.2)C(O)X,
wherein X is OH or an appropriate derivative or leaving group, in
the presence of conditions for coupling an amine and an acid (when
X is OH) or appropriate acid derivative (when X is not OH) to
provide the compound of formula I.
39. The process of claim 37, further comprising the step of
reacting the compound of formula I: ##STR00396## wherein Y is:
##STR00397## under hydrolysis conditions, to provide a compound of
formula II ##STR00398##
40. A process for preparing a compound of formula 3-A: ##STR00399##
wherein PG.sub.1 is a suitable carboxylic acid protecting group;
PG.sub.2 is a suitable nitrogen-protecting group; Ring A is:
##STR00400## wherein, in each ring, any hydrogen atom is optionally
and independently replaced by R.sup.4 and any set of two hydrogen
atoms bound to the same atom is optionally and independently
replaced by carbonyl; R.sup.4 is halogen, --OR.sup.9,
--NO.sub.2--CN --CF.sub.3, --OCF.sub.1, --R.sup.9,
1,2-methylenedioxy, 1,2-ethylenedioxy, --N(R.sup.9).sub.2,
--SR.sup.9, --SOR.sup.9,
--SO.sub.2R.sup.9--SO.sub.2N(R.sup.9).sub.2, --SO.sub.3R.sup.9,
--C(O)R.sup.9, --C(O)C(O)R.sup.9, --C(O)C(O)OR.sup.9,
--C(O)C(O)N(R.sup.9).sub.2, --C(O)CH.sub.2C(O)R.sup.9,
--C(S)R.sup.9, --C(S)OR.sup.9, --C(O)OR.sup.9, --OC(O)R.sup.9,
--C(O)N(R.sup.9).sub.2, --OC(O)N(R.sup.9).sub.2,
--C(S)N(R.sup.9).sub.2, --(CH.sub.2).sub.0-2NHC(O)R.sup.9,
--N(R.sup.9)N(R.sup.9)COR.sup.9,
--N(R.sup.9)N(R.sup.9)C(O)OR.sup.9,
--N(R.sup.9)N(R.sup.9)CON(R.sup.9).sub.2,
--N(R.sup.9)SO.sub.2R.sup.9, --N(R.sup.9)SO.sub.2N(R.sup.9).sub.2,
--N(R.sup.9)C(O)OR.sup.9, --N(R.sup.9)C(O)R.sup.9,
--N(R.sup.9)C(S)R.sup.9, --N(R.sup.9)C(O)N(R.sup.9).sub.2,
--N(R.sup.9)C(S)N(R.sup.9).sub.2, --N(COR.sup.9)COR.sup.9,
--N(OR.sup.9)R.sup.9, --C(.dbd.NH)N(R.sup.9).sub.2,
--C(O)N(OR.sup.9)R.sup.9, --C(.dbd.NOR.sup.9)R.sup.9,
--OP(O)(OR.sup.9).sub.2, --P(O)(R.sup.9).sub.2,
--P(O)(OR.sup.9).sub.2, or --P(O)(H)(OR.sup.9); R.sup.9 is
hydrogen, C.sub.1-12aliphatic, C.sub.3-10cycloaliphatic,
C.sub.6-10aryl, 5-10 membered heterocyclyl, 5-10 membered
heteroaryl, (C.sub.3-10cycloaliphatic)-(C.sub.1-12aliphatic)-,
(C.sub.6-10aryl)-(C.sub.1-12aliphatic)-, (5-10 membered
heterocyclyl)-(C.sub.1-12aliphatic), or
heteroaryl-(C.sub.1-12aliphatic)-; wherein any hydrogen atom is
optionally and independently replaced by R.sup.13 and any set of
two hydrogen atoms bound to the same atom is optionally and
independently replaced by carbonyl; comprising: reacting a compound
of formula 2-A: ##STR00401## wherein PG.sub.1 is as defined above;
and a compound of formula 20-A: ##STR00402## wherein PG.sub.2 and
Ring A are as defined above; X is OH or an appropriate leaving
group, under conditions for coupling an amine and a carboxylic acid
(when X is OH) or an amine and an appropriate carboxylic acid (when
X is an appropriate leaving group) to provide the compound of
formula 3-A.
41. The process of claim 40, wherein Ring A is ##STR00403##
42. The process of claim 40, wherein Ring A is ##STR00404##
43. A compound selected from the following: ##STR00405##
##STR00406## wherein PG.sub.1 is a suitable carboxylic acid
protecting group; PG.sub.2 is a suitable nitrogen-protecting group;
Z is a Z-type protecting group; R.sup.1 is H, C.sub.1-12aliphatic,
C.sub.3-10cycloaliphatic, C.sub.6-10aryl, 5-10 membered
heterocyclyl, 5-10 membered heteroaryl,
(C.sub.3-10cycloalkyl)-(C.sub.1-12aliphatic)-,
cycloalkenyl-(C.sub.1-12aliphatic)-,
(C.sub.6-10aryl)-(C.sub.1-12aliphatic)-, (5-10 membered
heterocyclyl)-(C.sub.1-12aliphatic)-, or (5-10 membered
heteroaryl)-(C.sub.1-12aliphatic)-, wherein any hydrogen atom is
optionally and independently replaced by R.sup.8 and any set of two
hydrogen atoms bound to the same atom is optionally and
independently replaced by carbonyl; Ring A is: ##STR00407##
wherein, in each ring, any hydrogen atom is optionally and
independently replaced by R.sup.4 and any set of two hydrogen atoms
bound to the same atom is optionally and independently replaced by
carbonyl; R.sup.4 is halogen, --OR.sup.9, --NO.sub.2--CN,
--CF.sub.3, --OCF.sub.1, --R.sup.9, 1,2-methylenedioxy,
1,2-ethylenedioxy, --N(R.sup.9).sub.2, --SR.sup.9,
--SO.sub.7R.sup.9--SO.sub.2N(R.sup.9).sub.2, --SO.sub.3R.sup.9,
--C(O)R.sup.9, --C(O)C(O)R.sup.9, --C(O)C(O)OR.sup.9,
--C(O)C(O)N(R.sup.9).sub.2, --C(O)CH.sub.2C(O)R.sup.9,
--C(S)R.sup.9, --C(S)OR.sup.9, --C(O)OR.sup.9, --OC(O)R.sup.9,
--C(O)N(R.sup.9).sub.2, --OC(O)N(R.sup.9).sub.2,
--C(S)N(R.sup.9).sub.2, --(CH.sub.2).sub.0-2NHC(O)R.sup.9,
--N(R.sup.9)N(R.sup.9)COR.sup.9,
--N(R.sup.9)N(R.sup.9)C(O)OR.sup.9,
--N(R.sup.9)N(R.sup.9)CON(R.sup.9).sub.2,
--N(R.sup.9)SO.sub.2R.sup.9, --N(R.sup.9)SO.sub.2N(R.sup.9).sub.2,
--N(R.sup.9)C(O)OR.sup.9, --N(R.sup.9)C(O)R.sup.9,
--N(R.sup.9)C(S)R.sup.9, --N(R.sup.9)C(O)N(R.sup.9).sub.2,
--N(R.sup.9)C(S)N(R.sup.9).sub.2, --N(COR.sup.9)COR.sup.9,
--N(OR.sup.9)R.sup.9, --C(.dbd.NH)N(R.sup.9).sub.2,
--C(O)N(OR.sup.9)R.sup.9, --C(.dbd.NOR.sup.9)R.sup.9,
--OP(O)(OR.sup.9).sub.2, --P(O)(R.sup.9).sub.2,
--P(O)(OR.sup.9).sub.2, or --P(O)(H)(OR.sup.9); R.sup.9 is
hydrogen, C.sub.1-12aliphatic, C.sub.3-10cycloaliphatic,
C.sub.6-10aryl, 5-10 membered heterocyclyl, 5-10 membered
heteroaryl, (C.sub.3-10cycloaliphatic)-(C.sub.1-12aliphatic)-,
(C.sub.6-10aryl)-(C.sub.1-12aliphatic)-, (5-10 membered
heterocyclyl)-(C.sub.1-12aliphatic)-, or
heteroaryl-(C.sub.1-12aliphatic)-; wherein any hydrogen atom is
optionally and independently replaced by R.sup.13 and any set of
two hydrogen atoms bound to the same atom is optionally and
independently replaced by carbonyl.
44-51. (canceled)
52. The process of claim 38, further comprising the step of
reacting the compound of ##STR00408## wherein Y is: ##STR00409##
under hydrolysis conditions, to provide a compound of formula II
##STR00410##
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This present application claims the benefit, under 35 U.S.C.
.sctn.119, to U.S. Provisional Application Nos. 60/548,610 filed
Feb. 27, 2004; 60/629,743 filed Nov. 19, 2004; and 60/629,661 filed
Nov. 19, 2004; the entire disclosures of which are incorporated
herein by reference.
FIELD OF THE INVENTION
[0002] This invention relates to compounds, and compositions
thereof, that are useful as caspase inhibitors.
[0003] This invention also relates to processes for preparing these
compounds.
[0004] This invention further relates to pharmaceutical
compositions comprising said compounds and to the use of the
compounds and compositions thereof for the treatment of diseases
and disorders related to caspase-mediated conditions.
BACKGROUND OF THE INVENTION
[0005] Caspases are a family of cysteine protease enzymes that are
key mediators in inflammation. Caspase-1 (ICE) processes
pre-IL-1.beta. to produce the active form of IL-1.beta. [WO
99/47545]. ICE has also been linked to the conversion of pro-IGIF
to IGIF and/or to the production of IFN-.gamma. [Id.]. Both
IL-1.beta. and IFN-.gamma. contribute to the pathology associated
with inflammatory, infectious, and autoimmune diseases (see, e.g.,
WO 99/47545; J. Invest. Dermatology, 120(1), pp. 164-167 (2003);
Br. J. Dermatology, 141, pp. 739-746 (1999); Science, 282, pp.
490-493 (1998); Schweiz. Med. Wochenschr., 130, pp. 1656-1661
(2000)].
[0006] Caspases are also key mediators in the signaling pathways
for apoptosis and cell disassembly [N. A. Thornberry, Chem. Biol.,
5, pp. R97-R103 (1998)]. 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.
[0007] 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 improve survival
after endotoxic shock [H. Yaoita et al., Circulation, 97, pp.
276-281 (1998); M. Endres et al., J. Cerebral Blood Flow and
Metabolism, 18, pp. 238-247, (1998); Y. Cheng et al., J. Clin.
Invest., 101, pp. 1992-1999 (1998); A. G. Yakovlev et al., J.
Neurosci., 17, pp. 7415-7424 (1997); I. Rodriquez et al., J. Exp.
Med., 184, pp. 2067-2072 (1996); Grobmyer et al., Mol. Med., 5, p.
585 (1999)].
[0008] However, due to their peptidic nature, such inhibitors are
typically characterized by undesirable pharmacological properties,
such as poor cellular penetration and cellular activity, poor oral
absorption, poor stability and rapid metabolism [J. J. Plattner and
D. W. Norbeck, in Drug Discovery Technologies, C. R. Clark and W.
H. Moos, Eds. (Ellis Horwood, Chichester, England, 1990), pp.
92-126]. This has hampered their development into effective drugs.
These and other studies with peptidic caspase inhibitors have
demonstrated that an aspartic acid residue is involved in a key
interaction with the caspase enzyme [K. P. Wilson et al., Nature,
370, pp. 270-275 (1994); Lazebnik et al., Nature, 371, p. 346
(1994)].
[0009] Accordingly, peptidyl and non-peptidyl aspartic acid
compounds are useful as caspase inhibitors.
[0010] A need nevertheless exists for compounds that have the
ability to act as caspase inhibitors, particularly with selective
activity against certain caspases.
SUMMARY OF THE INVENTION
[0011] The present invention provides a compound of formula I:
##STR00002##
wherein the variables are as defined herein.
[0012] The present invention also provides processes for preparing
these compounds, compositions, pharmaceutical compositions, and
methods of using such compounds and compositions for inhibiting
caspases. These compounds are particularly useful as selective
caspase-1/capase-8 inhibitors.
DETAILED DESCRIPTION OF THE INVENTION
[0013] The present invention provides a compound of formula I:
##STR00003##
wherein:
[0014] Y is
##STR00004##
[0015] R is R.sup.3C(O)--, HC(O), R.sup.3SO.sub.2--, R.sup.3OC(O),
(R.sup.3).sub.2NC(O), (R.sup.3) (H)NC(O), R.sup.3C(O)C(O)--,
R.sup.3--, (R.sup.3).sub.2NC(O)C(O), (R.sup.3) (H)NC(O)C(O), or
R.sup.3OC(O)C(O)--;
[0016] R.sup.1 is H, aliphatic, cycloaliphatic, aryl, heterocyclyl,
heteroaryl, cycloalkyl-aliphatic-, cycloalkenyl-aliphatic-,
aryl-aliphatic-, heterocyclyl-aliphatic-, or heteroaryl-aliphatic-,
wherein any hydrogen atom is optionally and independently replaced
by R.sup.8 and any set of two hydrogen atoms bound to the same atom
is optionally and independently replaced by carbonyl;
[0017] Ring A is:
##STR00005##
wherein, in each ring, any hydrogen atom is optionally and
independently replaced by R.sup.4 and any set of two hydrogen atoms
bound to the same atom is optionally and independently replaced by
carbonyl;
[0018] R.sup.3 is aliphatic, cycloaliphatic, aryl, heterocyclyl,
heteroaryl, cycloaliphatic-aliphatic-, aryl-aliphatic-,
heterocyclyl-aliphatic-, or heteroaryl-aliphatic-; or two R.sup.3
groups bound to the same atom form together with that atom a 3-10
membered aromatic or nonaromatic ring; wherein any ring is
optionally fused to an aryl, heteroaryl, cycloalkyl, or
heterocyclyl; wherein up to 3 aliphatic carbon atoms may be
replaced by a group selected from O, N, NR.sup.9, S, SO, and
SO.sub.2, wherein R.sup.3 is substituted with up to 6 substituents
independently selected from R.sup.8;
[0019] R.sup.4 is halogen, --OR.sup.9, --NO.sub.2, --CN,
--CF.sub.3, --OCF.sub.3, --R.sup.9, 1,2-methylenedioxy,
1,2-ethylenedioxy, --N(R.sup.9).sub.2, --SR.sup.9, --SOR.sup.9,
--SO.sub.2R.sup.9, --SO.sub.2N(R.sup.9).sub.2, --SO.sub.3R.sup.9,
--C(O)R.sup.9, --C(O)C(O)R.sup.9, --C(O)C(O)OR.sup.9,
--C(O)C(O)N(R.sup.9).sub.2, --C(O)CH.sub.2C(O)R.sup.9,
--C(S)R.sup.9, --C(S)OR.sup.9, --C(O)OR.sup.9, --OC(O)R.sup.9,
--C(O)N(R.sup.9).sub.2, --OC(O)N(R.sup.9).sub.2,
--C(S)N(R.sup.9).sub.2, --(CH.sub.2).sub.0-2NHC(O)R.sup.9,
--N(R.sup.9)N(R.sup.9)COR.sup.9,
--N(R.sup.9)N(R.sup.9)C(O)OR.sup.9,
--N(R.sup.9)N(R.sup.9)CON(R.sup.9).sub.2,
--N(R.sup.9)SO.sub.2R.sup.9, --N(R.sup.9)SO.sub.2N(R.sup.9).sub.2,
--N(R.sup.9)C(O)OR.sup.9, --N(R.sup.9)C(O)R.sup.9,
--N(R.sup.9)C(S)R.sup.9, --N(R.sup.9)C(O)N(R.sup.9).sub.2,
--N(R.sup.9)C(S)N(R.sup.9).sub.2, --N(COR.sup.9)COR.sup.9,
--N(OR.sup.9)R.sup.9, --C(.dbd.NH)N(R.sup.9).sub.2,
--C(O)N(OR.sup.9)R.sup.9, --C(.dbd.NOR.sup.9)R.sup.9,
--OP(O)(OR.sup.9).sub.2, --P(O)(R.sup.9).sub.2,
--P(O)(OR.sup.9).sub.2, or --P(O) (H)(OR.sup.9);
[0020] R.sup.2 is --C(R.sup.5)(R.sup.6)(R.sup.7), aryl, heteroaryl,
or C.sub.3-7 cycloalkyl;
[0021] R.sup.5 is H or a C.sub.1-6 straight-chained or branched
alkyl;
[0022] R.sup.6 is H or a C.sub.1-6 straight-chained or branched
alkyl;
[0023] R.sup.7 is --CF.sub.3, --C.sub.3-7cycloalkyl, aryl,
heteroaryl, heterocycle, or a C.sub.1-6 straight-chained or
branched alkyl, wherein each carbon atom of the alkyl is optionally
and independently substituted with R.sup.10;
[0024] Or R.sup.5 and R.sup.7 taken together with the carbon atom
to which they are attached form a 3-10 membered cycloaliphatic;
[0025] R.sup.8 and R.sup.8' are each independently halogen,
--OR.sup.9, --NO.sub.2, --CN, --CF.sub.3, --OCF.sub.3, --R.sup.9,
1,2-methylenedioxy, 1,2-ethylenedioxy, --N(R.sup.9).sub.2,
--SR.sup.9, --SOR.sup.9, --SO.sub.2R.sup.9,
--SO.sub.2N(R.sup.9).sub.2, --SO.sub.3R.sup.9, --C(O)R.sup.9,
--C(O)C(O)R.sup.9, --C(O)C(O)OR.sup.9, --C(O)C(O)N(R.sup.9).sub.2,
--C(O)CH.sub.2C(O)R.sup.9, --C(S)R.sup.9, --C(S)OR.sup.9,
--C(O)OR.sup.9, --OC(O)R.sup.9, --C(O)N(R.sup.9).sub.2,
--OC(O)N(R.sup.9).sub.2, --C(S)N(R.sup.9).sub.2,
--(CH.sub.2).sub.0-2NHC(O)R.sup.9, --N(R.sup.9)N(R.sup.9)COR.sup.9,
--N(R.sup.9)N(R.sup.9)C(O)OR.sup.9,
--N(R.sup.9)N(R.sup.9)CON(R.sup.9).sub.2,
--N(R.sup.9)SO.sub.2R.sup.9, --N(R.sup.9)SO.sub.2N(R.sup.9).sub.2,
--N(R.sup.9)C(O)OR.sup.9, --N(R.sup.9)C(O)R.sup.9,
--N(R.sup.9)C(S)R.sup.9, --N(R.sup.9)C(O)N(R.sup.9).sub.2,
--N(R.sup.9)C(S)N(R.sup.9).sub.2, --N(COR.sup.9)COR.sup.9,
--N(OR.sup.9)R.sup.9, --C(.dbd.NH)N(R.sup.9).sub.2,
--C(O)N(OR.sup.9)R.sup.9, --C(.dbd.NOR.sup.9)R.sup.9,
--OP(O)(OR.sup.9).sub.2, --P(O)(R.sup.9).sub.2,
--P(O)(OR.sup.9).sub.2, and --P(O) (H)(OR.sup.9);
[0026] R.sup.9 is hydrogen, aliphatic, cycloaliphatic, aryl,
heterocyclyl, heteroaryl, cycloaliphatic-aliphatic-,
aryl-aliphatic-, heterocyclyl-aliphatic-, or heteroaryl-aliphatic-;
wherein any hydrogen atom is optionally and independently replaced
by R.sup.8 and any set of two hydrogen atoms bound to the same atom
is optionally and independently replaced by carbonyl;
[0027] R.sup.10 is halogen, --OR.sup.11, --NO.sub.2, --CN,
--CF.sub.3, --OCF.sub.3, --R.sup.11, or --SR.sup.11; wherein
R.sup.11 is C.sub.1-4-aliphatic-.
[0028] The present invention also provides a compound of formula
II:
##STR00006##
wherein:
[0029] Y is
##STR00007##
[0030] R.sup.1 is H, aliphatic, cycloalkyl (e.g., cyclopentyl),
cycloalkenyl, aryl, heterocyclyl, heteroaryl,
cycloalkyl-aliphatic-cycloalkenyl-aliphatic-, aryl-aliphatic-,
heterocyclyl-aliphatic-, or heteroaryl-aliphatic-, wherein any
hydrogen atom is optionally and independently replaced by R.sup.8
and any set of two hydrogen atoms bound to the same atom is
optionally and independently replaced by carbonyl;
[0031] Ring A is:
##STR00008##
wherein, in each ring, any hydrogen atom is optionally and
independently replaced by R.sup.4 and any set of two hydrogen atoms
bound to the same atom is optionally and independently replaced by
carbonyl (or in an alternative embodiment, carbonyl or (C3-C6)
spirocycle;)
[0032] R.sup.4 is halogen, --OR.sup.9, --NO.sub.2, --CN,
--CF.sub.2, --OCF.sub.2, --R.sup.9, 1,2-methylenedioxy,
1,2-ethylenedioxy, --N(R.sup.9).sub.2, --SR.sup.9, --SOR.sup.9,
--SO.sub.2R.sup.9, --SO.sub.2N(R.sup.9).sub.2, --SO.sub.3R.sup.9,
--C(O)R.sup.9, --C(O)C(O)R.sup.9, --C(O)C(O)OR.sup.9,
--C(O)C(O)N(R.sup.9).sub.2, --C(O)CH.sub.2C(O)R.sup.9,
--C(S)R.sup.9, --C(S)OR.sup.9, --C(O)OR.sup.9, --OC(O)R.sup.9,
--C(O)N(R.sup.9).sub.2, --OC(O)N(R.sup.9).sub.2,
--C(S)N(R.sup.9).sub.2, --(CH.sub.2).sub.0-2NHC(O)R.sup.9,
--N(R.sup.9)N(R.sup.9)COR.sup.9,
--N(R.sup.9)N(R.sup.9)C(O)OR.sup.9,
--N(R.sup.9)N(R.sup.9)CON(R.sup.9).sub.2,
--N(R.sup.9)SO.sub.2R.sup.9, --N(R.sup.9)SO.sub.2N(R.sup.9).sub.2,
--N(R.sup.9)C(O)OR.sup.9, --N(R.sup.9)C(O)R.sup.9,
--N(R.sup.2)C(S)R.sup.9, --N(R.sup.9)C(O)N(R.sup.9).sub.2,
--N(R.sup.9)C(S)N(R.sup.9).sub.2, --N(COR.sup.9)COR.sup.9,
--N(OR.sup.9)R.sup.9, --C(.dbd.NH)N(R.sup.9).sub.2,
--C(O)N(OR.sup.9)R.sup.9, --C(.dbd.NOR.sup.9)R.sup.9,
--OP(O)(OR.sup.9).sub.2, --P(O)(R.sup.9).sub.2,
--P(O)(OR.sup.9).sub.2, or --P(O) (H)(OR.sup.9);
[0033] R.sup.2 is --C(R.sup.5)(R.sup.6)(R.sup.7), aryl, heteroaryl,
or --C.sub.3-7 cycloalkyl;
[0034] R.sup.5 is H or a C.sub.1-6 straight-chained or branched
alkyl;
[0035] R.sup.6 is H or a C.sub.1-6 straight-chained or branched
alkyl;
[0036] R.sup.7 is --CF.sub.3, --C.sub.3-7 cycloalkyl, aryl,
heteroaryl, heterocycle, or a C.sub.1-6 straight-chained or
branched alkyl, wherein each carbon atom of the alkyl is optionally
and independently substituted with R.sup.10;
[0037] (or in an alternative embodiment, R.sup.5 and R.sup.7 taken
together with the carbon atom to which they are attached form a
3-10 membered cycloaliphatic);
[0038] R.sup.3 is phenyl, thiophene, or pyridine, wherein each ring
is optionally substituted with up to 5 groups independently
selected from R.sup.8', and wherein at least one position on the
phenyl, thiophene, or pyridine adjacent to bond x is substituted by
R.sup.12, wherein R.sup.12 has no more than 5 straight-chained
atoms;
[0039] R.sup.8 and R.sup.8' are each independently halogen,
--OR.sup.9, --NO.sub.2, --CN, --CF.sub.3, --OCF.sub.3, --R.sup.9,
1,2-methylenedioxy, 1,2-ethylenedioxy, --N(R.sup.9).sub.2,
--SR.sup.9, --SOR.sup.9, --SO.sub.2R.sup.9,
--SO.sub.2N(R.sup.9).sub.2, --SO.sub.3R.sup.9, --C(O)R.sup.9,
--C(O)C(O)R.sup.9, --C(O)C(O)OR.sup.9, --C(O)C(O)N(R.sup.9).sub.2,
--C(O)CH.sub.2C(O)R.sup.9, --C(S)R.sup.9, --C(S)OR.sup.9,
--C(O)OR.sup.9, --OC(O)R.sup.9, --C(O)N(R.sup.9).sub.2,
--OC(O)N(R.sup.9).sub.2, --C(S)N(R.sup.9).sub.2,
--(CH.sub.2).sub.0-2NHC(O)R.sup.9, --N(R.sup.9)N(R.sup.9)COR.sup.9,
--N(R.sup.9)N(R.sup.9)C(O)OR.sup.9,
--N(R.sup.9)N(R.sup.9)CON(R.sup.9).sub.2,
--N(R.sup.9)SO.sub.2R.sup.9, --N(R.sup.9)SO.sub.2N(R.sup.9).sub.2,
--N(R.sup.9)C(O)OR.sup.9, --N(R.sup.9)C(O)R.sup.9,
--N(R.sup.9)C(S)R.sup.9, --N(R.sup.9)C(O)N(R.sup.9).sub.2,
--N(R.sup.9)C(S)N(R.sup.9).sub.2, --N(COR.sup.9)COR.sup.9,
--N(OR.sup.9)R.sup.9, --C(.dbd.NH)N(R.sup.9).sub.2,
--C(O)N(OR.sup.9)R.sup.9, --C(.dbd.NOR.sup.9)R.sup.9,
--OP(O)(OR.sup.9).sub.2, --P(O)(R.sup.9).sub.2,
--P(O)(OR.sup.9).sub.2, and --P(O) (H)(OR.sup.9);
[0040] R.sup.9 is hydrogen, aliphatic, cycloalkyl, cycloalkenyl,
aryl, heterocyclyl, heteroaryl, cycloaliphatic-aliphatic-,
aryl-aliphatic-, heterocyclyl-aliphatic-, or heteroaryl-aliphatic-;
(in certain embodiments, any hydrogen atom of R.sup.9 is optionally
and independently replaced by R.sup.8 and any set of two hydrogen
atoms bound to the same atom is optionally and independently
replaced by carbonyl; provided that if R.sup.9 is substituted with
a R.sup.8, wherein the R.sup.8 comprises a R.sup.9 substituent,
then that R.sup.9 substituent is not substituted with R.sup.8);
[0041] R.sup.10 is halogen, --OR.sup.11, --NO.sub.2, --CN,
--CF.sub.3, --OCF.sub.3, --R.sup.11, or --SR.sup.11;
[0042] R.sup.11 is C.sub.1-4-aliphatic-; and
[0043] R.sup.12 is halogen, --OR.sup.11, --NO.sub.2, --CN,
--CF.sub.3, --OCF.sub.3, --R.sup.11, --SR.sup.9.
[0044] As used in the definition of R.sup.12, "straight-chained
atoms" refers to atoms that are linearly bound, regardless of
whether those atoms also have atoms bound in a branched fashion.
According to this definition, an ethyl group and a trifluoromethoxy
group each have three straight-chained atoms, and a methyl group
has two straight-chained atoms. In the above embodiment, R.sup.12
has no more than 5 straight-chained atoms. In two other
embodiments, R.sup.12 has no more than 4 straight-chained atoms and
no more than 3 straight-chained atoms. In yet other embodiments,
R.sup.12 has 2 straight-chained atoms or 1 atom.
[0045] As used herein, a position adjacent to the bond x refers to
a position which is located next to the position at which x is
bound. In an aryl ring, this position is often called "the ortho
position" or, in the case of a phenyl ring, it may be called "the
2-position". By way of example, in the structures immediately
below, R.sup.12 is bound to the phenyl, thiophene, and pyridine
rings at "the position adjacent to bond x".
##STR00009##
[0046] In one embodiment of this invention, R is R.sup.3C(O)--.
[0047] In some embodiments, R.sup.3 is optionally substituted
C.sub.6-10aryl or heteroaryl. In other embodiments R.sup.3 is
optionally substituted phenyl. In yet other embodiments, R.sup.3 is
a 8-10 membered optionally substituted heteroaryl (i.e. quinoline,
isoquinoline, or quinazoline) In yet other embodiments, R.sup.3 is
an optionally substituted 5-6 membered heteroaryl (i.e., pyridyl,
pyrimidyl, pyrazinyl, thiophenyl, furanyl, thiazolyl).
[0048] In some embodiments, R.sup.3 is optionally and independently
substituted by 0-5 R.sup.8' groups.
[0049] In one embodiment, the compound of this invention is
represented by formula II:
##STR00010##
wherein: [0050] a)R.sup.3 is phenyl, thiophene, or pyridine; [0051]
b) each ring is optionally substituted with up to 5 groups
independently selected from R.sup.8'; and [0052] c) at least one
position on the phenyl, thiophene, or pyridine adjacent to bond x
is substituted by R.sup.12, wherein R.sup.12 has no more than 5
straight-chained atoms.
[0053] Another embodiment of this invention provides a compound
wherein Y is:
##STR00011##
[0054] In one embodiment of this invention, R.sup.1 is substituted
with up to 3 groups selected independently from carbonyl and
R.sup.8.
[0055] In another embodiment, R.sup.1 is C.sub.1-12aliphatic or
C.sub.3-10cycloalkyl, wherein each R.sup.1 is optionally
substituted with 1-3 groups selected independently from R.sup.8. In
yet another embodiment, R.sup.1 is a straight-chain or branched
C.sub.1-4 alkyl that is optionally substituted with 1-3 groups
selected independently from R.sup.8.
[0056] In one embodiment, R.sup.1 is an unsubstituted,
straight-chain or branched C.sub.1-4 alkyl (e.g., ethyl, isopropyl,
n-propyl, or n-butyl). In another embodiment, R.sup.1 is ethyl.
[0057] In any of these embodiments, R.sup.8 is halogen, --OR.sup.9,
--CN, --CF.sub.3, --OCF.sub.3, or --R.sup.9. In another embodiment
wherein R.sup.8 is --R.sup.9, that R.sup.9 is benzyl.
[0058] In another embodiment, Y is
##STR00012##
[0059] In another embodiment, Ring A is substituted with up to 3
groups (preferably, 1 group) selected independently from carbonyl
and R.sup.4.
[0060] In one embodiment, Ring A is:
##STR00013##
optionally substituted with R.sup.4.
[0061] In yet another embodiment, Ring A is:
##STR00014##
optionally substituted with R.sup.4.
[0062] In another form of this embodiment, Ring A is unsubstituted
proline (i.e., R.sup.4 is hydrogen).
[0063] In yet another embodiment, Ring A is:
##STR00015##
optionally substituted with R.sup.4.
[0064] In one embodiment, Ring A is
##STR00016##
optionally substituted with R.sup.4.
[0065] In any of these embodiments, R.sup.4 is halogen, --OR.sup.9,
--CF.sub.3, --OCF.sub.3, --R.sup.9, or --SR.sup.9. In certain
embodiments R.sup.4 is H.
[0066] In one embodiment, R.sup.2 is a C.sub.3-4 branched alkyl
group.
[0067] In another embodiment, R.sup.5 is H or --CH.sub.3, R.sup.6
is --CH.sub.3, and R.sup.7 is --CH.sub.3.
[0068] In another embodiment, R.sup.12 is --OCF.sub.3, --OCH.sub.3,
--CF.sub.3, --CH.sub.3, --CH.sub.2CH.sub.3, --Cl, or --F.
[0069] In yet another embodiment, R.sup.12 is --CF.sub.3,
--CH.sub.3, --Cl, or --F.
[0070] In yet another embodiment, R.sup.12 is --CH.sub.3, --Cl, or
--F.
[0071] In another embodiment, each R.sup.8', if present, is
independently halogen, --OR.sup.9NO.sub.2, --CN, --CF.sub.3,
--OCF.sub.3, --R.sup.9, 1,2-methylenedioxy, 1,2-ethylenedioxy,
--N(R.sup.9).sub.2, --SR.sup.9, --SOR.sup.9, --SO.sub.2R.sup.9,
--SO.sub.2N(R.sup.9).sub.2, --C(O)R.sup.9,
--C(O)C(O)N(R.sup.9).sub.2, --C(O)N(R.sup.9).sub.2,
--OC(O)N(R.sup.9).sub.2, --(CH.sub.2).sub.0-2NHC(O)R.sup.9,
--N(R.sup.9)SO.sub.2R.sup.9, --N(R.sup.9)SO.sub.2N(R.sup.9).sub.2,
--N(R.sup.9)C(O)OR.sup.9, --N(R.sup.9)C(O)R.sup.9, or
--N(R.sup.9)C(O)N(R.sup.9).sub.2.
[0072] In another embodiment, R.sup.8' is --NH.sub.2,
--N(R.sup.9).sub.2, --N(R.sup.9)C(O)R.sup.9, --OCF.sub.3,
--OR.sup.9, --CF.sub.3, --R.sup.9, --SR.sup.9 or halo. In this
embodiment, halo is, preferably, Cl or F and R.sup.9 is,
preferably, straight or branched C.sub.1-4 alkyl.
[0073] According to one embodiment, this invention provides
compounds of formula III:
##STR00017##
wherein the variables are as defined in any of the embodiments
herein.
[0074] In one form of this embodiment, the compound has the
stereochemistry indicated below:
##STR00018##
wherein the variables are as defined in any of the embodiments
herein.
[0075] In other forms of this embodiment, the compound has the
stereochemistry indicated below:
##STR00019##
wherein the variables are as defined in any of the embodiments
herein.
[0076] According to another embodiment, this invention provides
compound of formula IV:
##STR00020##
wherein the variables are as defined in any of the embodiments
herein.
[0077] In one form of this embodiment, the compound has the
stereochemistry indicated below:
##STR00021##
wherein the variables are as defined in any of the embodiments
herein.
[0078] The embodiments herein may be combined to provide a compound
according to this invention.
[0079] According to one embodiment, the present invention provides
a compound selected from Table 1 below:
TABLE-US-00001 TABLE 1 ##STR00022## ##STR00023## ##STR00024##
##STR00025## ##STR00026## ##STR00027## ##STR00028## ##STR00029##
##STR00030## ##STR00031## ##STR00032## ##STR00033## ##STR00034##
##STR00035## ##STR00036## ##STR00037## ##STR00038## ##STR00039##
##STR00040## ##STR00041## ##STR00042## ##STR00043## ##STR00044##
##STR00045## ##STR00046## ##STR00047## ##STR00048## ##STR00049##
##STR00050## ##STR00051## ##STR00052## ##STR00053## ##STR00054##
##STR00055## ##STR00056## ##STR00057## ##STR00058## ##STR00059##
##STR00060## ##STR00061## ##STR00062## ##STR00063## ##STR00064##
##STR00065## ##STR00066## ##STR00067## ##STR00068## ##STR00069##
##STR00070## ##STR00071## ##STR00072## ##STR00073## ##STR00074##
##STR00075## ##STR00076## ##STR00077## ##STR00078## ##STR00079##
##STR00080## ##STR00081## ##STR00082## ##STR00083## ##STR00084##
##STR00085## ##STR00086## ##STR00087## ##STR00088## ##STR00089##
##STR00090## ##STR00091## ##STR00092## ##STR00093##
##STR00094##
[0080] According to another embodiment, the present invention
provides a compound of formula II selected from Table 2 below:
TABLE-US-00002 TABLE 2 ##STR00095## ##STR00096## ##STR00097##
##STR00098## ##STR00099## ##STR00100## ##STR00101## ##STR00102##
##STR00103## ##STR00104## ##STR00105## ##STR00106## ##STR00107##
##STR00108## ##STR00109## ##STR00110## ##STR00111## ##STR00112##
##STR00113## ##STR00114## ##STR00115## ##STR00116## ##STR00117##
##STR00118## ##STR00119## ##STR00120## ##STR00121## ##STR00122##
##STR00123## ##STR00124## ##STR00125## ##STR00126## ##STR00127##
##STR00128## ##STR00129## ##STR00130## ##STR00131## ##STR00132##
##STR00133## ##STR00134## ##STR00135## ##STR00136## ##STR00137##
##STR00138## ##STR00139## ##STR00140## ##STR00141## ##STR00142##
##STR00143## ##STR00144## ##STR00145## ##STR00146## ##STR00147##
##STR00148## ##STR00149## ##STR00150## ##STR00151## ##STR00152##
##STR00153## ##STR00154## ##STR00155## ##STR00156## ##STR00157##
##STR00158## ##STR00159## ##STR00160##
[0081] In certain embodiments of this invention, the variable
definitions are selected from those depicted in the compounds of
Table 1 and/or Table 2.
[0082] As used herein, a specified number atoms includes any
integer therein. For example, a group having from 1-4 atoms, could
have 1, 2, 3, or 4 atoms.
[0083] 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).
[0084] Cycloalkyl and cycloalkenyl groups have between 3 and 10
carbon atoms and are monocyclic or bicyclic, including linearly
fused, bridged, or spirocyclic.
[0085] As used herein, "aromatic group" or "aryl" refers to a
6-10-membered ring system that contains at least one aromatic ring.
Examples of aromatic rings include phenyl and naphthyl.
[0086] 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.sup.9), O, S, SO, and SO.sub.2, wherein at least one ring is
heteroaromatic (e.g., pyridyl, thiophene, or thiazole).
[0087] 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.sup.9), O, S, SO, and SO.sub.2, wherein no ring is aromatic
(e.g., piperidine and morpholine).
[0088] Further 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).
[0089] Further 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.
[0090] Each of the above aliphatic, aryl, cycloaliphatic,
heteroaryl, and heterocyclyl may contain appropriate substituents
(preferably up to 5) independently selected from, for example,
carbonyl and R.sup.8. Preferred substituents are halogen,
--OR.sup.9, --NO.sub.2, --CF.sub.3, --OCF.sub.3, --R.sup.9, oxo,
--OR.sup.9, --O-benzyl, --O-phenyl, 1,2-methylenedioxy,
1,2-ethylenedioxy, --N(R.sup.9).sub.2, --C(O)R.sup.8, --COOR.sup.9
or --CON(R.sup.9).sub.2, wherein R.sup.9 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.
[0091] 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.
[0092] 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.
[0093] The compounds of this invention may be obtained by any
method, including general, synthetic methods known to those skilled
in the art for analogous compounds (see e.g., WO 99/47545). For the
purposes of illustration, the following Schemes for the synthesis
of the compounds of the present invention are provided.
[0094] 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 DIPEA is diisopropylethylamine DMF is
dimethylformamide TFA is trifluoroacetic acid Z is
benzyloxycarbonyl .sup.1H NMR is nuclear magnetic resonance TLC is
thin layer chromatography
##STR00161##
[0095] Scheme I depicts a general route to prepare the compounds E
and F disclosed in this invention. The amino group of species A,
readily obtained from reduction of the .alpha.-carboxylic group of
aspartic acid (protected with PG.sub.1 as an ester), is coupled to
the carboxylic acid moiety of species B (N-protected with PG.sub.2)
to give species C. PG.sub.1 and PG.sub.2 are orthogonal protecting
groups (i.e., protecting groups where a protecting group may be
selectively removed in the presence of another protecting group.
Ideally, PG.sub.1 should be able to be removed without removing
PG.sub.2 and vice versa). Here, the aspartate part of the molecule
is then manipulated in an
oxidation/ketalisation/deprotection/cyclisation sequence to give
species D. The Ring A portion of D is then functionalized further
to give species E which is part of the disclosed invention.
Deprotection of the ketal gives species F which represent the other
part of the disclosed invention.
[0096] In various embodiments of this invention, PG.sub.2 is a
suitable amine protecting group, including but not limited to, the
amine protecting groups described in T. W. Greene & P. G. M
Wutz, "Protective Groups in Organic Synthesis", 3.sup.rd Edition,
John Wiley & Sons, Inc. (1999 and other editions) ("Greene"). A
"Z" protecting group (benzyloxycarbonyl) is a particularly useful
N-protecting group for use in connection with this invention. In
compounds wherein PG.sub.2 is protecting the nitrogen of a proline,
PG.sub.2 is preferably Z. It should be understood that modified Z
groups ("Z-type protecting groups") employed in connection with the
compounds and processes of this invention would also fall within
the scope of this invention. For example, Z could be substituted at
the CH.sub.2 group or the phenyl group with R.sup.8 (preferably
halo or C.sub.1-6 straight-chained or branched alkyl) to provide a
Z-type protecting group.
[0097] In various embodiments of this invention, PG.sub.1 is a
suitable carboxylic acid protecting group, including but not
limited to the acid protecting groups described in Greene. In
certain embodiments, PG.sub.1 is C.sub.1-6 straight-chained or
branched alkyl group. A t-butyl group is a particularly useful acid
protecting group for use in connection with this invention.
[0098] In Scheme I, compound A is a modified aspartic acid residue.
In addition to compound A, other modified aspartic acid residues,
including the following, have been reported:
##STR00162##
wherein, PG.sub.3 and PG.sub.4 are appropriate protecting groups.
These modified aspartic acids may be prepared by methods known to
skilled practitioners. See, for example, United States Patent
Application Publication US 2002/0042376 (especially page 9,
paragraph [0121] and pages 21-22, paragraph [0250] and the
documents cited at paragraph [0123]) and U.S. Pat. No. 6,235,899.
See also, C. Gros et al. "Stereochemical control in the preparation
of .alpha.-amino N-methylthiazolidine Masked Aldehydes used for
Peptide Aldehyde Synthesis" Tetrahedron, 58, pp. 2673-2680 (2002);
K. T. Chapman, "Synthesis of a Potent Reversible Inhibitor of
Interleukin-.beta. Converting Enzyme" Bioorg. Med. Chem. Letts., 2,
pp. 613-618 (1982); M. D. Mullican et al. "The Synthesis and
Evaluation of Peptidyl Aspartyl Aldehydes as Inhibitors of ICE'" 4,
pp. 2359-2364 (1994); M. H. Chen, et al. "An Efficient
Stereoselective Synthesis of
[3S(1S,9S)]-3-[[[9-(Benzoylamino)octahydro-6,10-Dioxo-6H-pyridazino-(1,2--
a)(1,2)-Diazepin-1-yl]-carbonyl]amino]-4-oxobutanoic acid, an
interleukin converting enzyme (ICE) Inhibitor" 9, pp. 1587-1592
(1999). Accordingly, Scheme I (and also Scheme III below) could be
modified to use these other aspartic acid residues.
##STR00163##
[0099] Scheme II depicts formation of compounds of formula I and
II, wherein Ring A is unsubstituted proline. Here the cyclic acetal
form of a compound of this invention is depicted as formula I and
the aldehyde form is depicted as formula II. Compounds having a
Ring A other than unsubstituted proline could be substituted in the
methods depicted in Scheme I.
[0100] Scheme II depicts the routes utilized to prepare compounds
of formulae I and II. Compounds I can be prepared from compounds 1
by condensation of the amino group in 1 with the suitably
functionalized carboxylic acid (or derivative). In this step,
standard coupling reagents to form amide bonds have been depicted;
other conditions known in the art to form amide bonds can also be
used.
[0101] As known to skilled practitioners, a carboxylic acid
(--C(O)OH) can be coupled to the amine under appropriate conditions
for coupling amines and carboxylic acids. Alternatively, in such
couplings, a carboxylic acid derivative (--C(O)X) may be employed
instead of the carboxylic acid. It should be understood that in the
context of coupling an amine and a carboxylic acid derivative, the
derivative would activate the acid to facilitate coupling to an
amine. Appropriate X groups are essentially leaving groups and are
known to skilled practitioners. "March's Advanced Organic
Chemistry", 5.sup.th Ed., Ed.: Smith, M. B. and March, J., John
Wiley & Sons, New York: 2001.
[0102] Typical conditions for coupling an amine and an acid include
combining a suitable solvent, a carboxylic acid, a base, and a
peptide-coupling reagent. Examples of suitable conditions are
described in US2002/0042376 and WO 01/81330, the entireties of
which are hereby incorporated by reference. In certain embodiments,
the conditions are as described in the Schemes and Examples
herein.
[0103] Examples of appropriate derivatives include, but are not
limited to, compounds of the formula RX wherein X is Cl, F,
OC(.dbd.O)R'' (R'' is aliphatic or aryl), SH, SR, SAr, or SeAr. In
some embodiments R is C(.dbd.O). Suitable conditions for using
these appropriate derivatives are known in the art.
##STR00164##
[0104] Scheme III depicts a possible route to prepare compounds 7
and compounds 1 described in scheme I. Compound 2, readily obtained
from reduction of the .alpha.-carboxylic group of aspartic acid, is
coupled to N-protected proline (or other ring, wherein Ring A is
other than unsubstituted proline) to form 3. Here, the proline is
N-protected with a Z (benzyloxycarbonyl) group. Compounds 3 are
then oxidized into the aldehydes 4 which are acetalized in situ to
give the acetals 5. Acetals can be formed in the presence of
R.sup.1--OH (or a suitable acetal forming reagent), a protic acid
(for example, TsOH), or a Lewis acid, and a suitable solvent.
Examples of suitable acetal forming reagents that form compounds
wherein R.sup.1 is ethyl can be considered ethanol equivalents and
include, but are not limited to, triethylorthoformate or a
diethylacetal, such as a
(CH.sub.3).sub.2C(OCH.sub.2CH.sub.3).sub.2. Preferably, the solvent
is CH.sub.2Cl.sub.2, toluene, or chlorobenzene. Appropriate protic
acids include, but are not limited to, TFA, p-TsOH. Appropriate
Lewis acids include, but are not limited to TiCl.sub.4, MgBr.sub.2,
and ZnCl.sub.2.
[0105] In Scheme III, the oxidation of compounds 3 to compounds 4
is depicted as being done under Swern conditions. Other oxidation
conditions may also be employed to prepare compounds of this
invention. Preferred oxidation conditions are those that a mild and
relatively quick to minimize epimerization at the acid side chain
of the modified aspartic acid residue. In one embodiment, the
oxidation step is a TEMPO oxidation (see Example I-1, Method C,
below). Other oxidation conditions include a Dess-Martin oxidation
and a tetrapropylammonium perruthenate (TPAP) oxidation.
[0106] Aldehydes 4 may be isolated but are preferably carried
through directly to 5 without isolation. Deprotection of the
tert-butyl ester (in 5) is accompanied by spontaneous ring
cyclization to give a mixture of diastereoisomers which were
separated by column chromatography to give the enantiomerically
pure syn ketals 6 and anti ketals (not represented in this scheme).
The deprotection may be done under protic acid or Lewis acid
conditions in an appropriate solvent. Appropriate solvents include,
but are not limited to, toluene, chlorobenzene, and DCM.
Appropriate protic acids include, but are not limited to, TFA,
p-TsOH. Appropriate Lewis acids include, but are not limited to
TiCl.sub.4, MgBr.sub.2, and ZnCl.sub.2. For clarity of the scheme,
only syn ketals are represented in the next steps to form compounds
7 and 1 but the same sequence may be used to form anti ketals.
Compounds 6 are submitted to hydrogenolysis and the resulting
compounds 7 are reacted with Z-protected aminoacids, using
conditions known in the art to prepare amide bonds, to yield
compounds 9. Compounds 7 may be generated and used in situ. If
isolated, it is preferable to use compounds 7 relatively soon after
generation. Compounds 9 are finally submitted to hydrogenolysis to
give compounds I, which can be used directly to prepare compounds
I, as depicted in Scheme II.
[0107] Alternatively, compounds 7 can be used to prepare compounds
I, as depicted in Scheme II. In this preparation, an amino acid
residue and the desired N-terminal group is prepared in one step
(see, Scheme II, reaction (b)).
[0108] As described in connection with Scheme I, aspartic acid
derivatives other than compounds 2 can be employed to obtain
compounds of this invention.
##STR00165##
[0109] Scheme IV depicts formation of compounds of formula III and
IV, wherein Ring A is 2-Aza-bicyclo[2.2.1]-heptane-3-carboxylic
acid. Here the cyclic acetal form of a compound of this invention
is depicted as formula III and the aldehyde form is depicted as
formula IV. Scheme IV depicts the routes utilized to prepare
compounds of formulae III and IV. Compounds III can be prepared
from compounds 11 by condensation of the amino group in 11 under
conditions to provide the desired R group, such as suitably
functionalized carboxylic acid (or derivative), sulfonic acid (or
derivative), chloroformate or carbamoyl chloride (or isocyanate),
for example, under appropriate reaction condition. In this step,
standard coupling reagents to form CO--NH bonds have been depicted;
other conditions known in the art to form CO--NH (or alkyl-N, or
SO.sub.2--N) bonds can also be used to provide the desired compound
comprising R--N. Alternatively, compounds I can be prepared from
compounds 17 by condensation of the amino group in 17 with the
suitably functionalized carboxylic acid (or derivative), sulfonic
acid (or derivative), chloroformate or carbamoyl chloride (or
isocyanate). In this step, standard coupling reagents to form
CO--NH bonds have been depicted; other conditions known in the art
to form CO--NH bonds can also be used.
##STR00166## ##STR00167##
[0110] Scheme V depicts a possible route to prepare compounds 17
and compounds 11 described in scheme III. Compound 2, readily
obtained from reduction of the .alpha.-carboxylic group of aspartic
acid, is coupled to N-protected
2-aza-bicyclo[2.2.1]heptane-3-carboxylic acid 10 (prepared as in
Tetrahedron: Asymmetry, 13, 2002, 25-28) to form 13. Compound 13 is
then oxidized into the aldehyde 14 which is acetalized in situ to
give the acetals 15. Deprotection of the tert-butyl ester is
accompanied by spontaneous ring cyclization to give a mixture of
diastereoisomers which were separated by column chromatography to
give the enantiomerically pure syn ketals 16 and anti ketals (not
represented in this scheme). Alternative Ring A groups are either
commercially available, reported in the literature, or may be
prepared according to methods known in the literature.
[0111] For clarity of the scheme, only syn ketals are represented
in the next steps to form compounds 17 and 11 but the same sequence
may be used to form anti ketals. Compounds 16 are submitted to
hydrogenolysis and the resulting compounds 17 are reacted with
Z-protected aminoacids, using conditions known in the art to
prepare amide bonds, to yield compounds 19.
[0112] Alternatively, compounds 17 can be used to prepare compounds
III, as depicted in Scheme IV. Compounds 19 are finally submitted
to hydrogenolysis to give compounds 11, which can be used directly
to prepare compounds III, as depicted in Scheme IV.
[0113] The R.sup.3COOH used in Scheme II are either commercially
available, reported in the literature, or prepared according to
methods known in the literature. For compound II-30,
2-chloro-3-methoxybenzoic acid was prepared as in J. Org. Chem.,
59, 1994, 2939-2944.
[0114] For compound II-32, 2-chloro-3-trifluoromethoxybenzoic acid
was prepared from 2-amino-3-trifluoromethoxybenzoic acid (prepared
as in J. Org. Chem., 68, 2003, 4693-4699) using a Sandmeyer
replacement of the amino group by a chloro, according to a method
substantially similar to the one reported in J. Org. Chem., 59,
1994, 2939-2944.
[0115] Accordingly, this invention also provides a process for
preparing a compound of this invention.
[0116] In one embodiment is provided a process for preparing a
compound of formula I:
##STR00168##
[0117] wherein Y is:
##STR00169##
and the other variables are as defined in any of the embodiments
herein;
[0118] comprising reacting a compound of formula 1:
##STR00170##
wherein the variables are as defined in any of the embodiments
herein; and a compound of formula RX, wherein X is OH or an
appropriate derivative (i.e., leaving group), in the presence of
conditions for coupling an amine and an acid (when X is OH) or an
amine and an appropriate acid derivative (when X is not OH (i.e., a
leaving group; for example, Cl) to provide the compound of formula
I.
[0119] Another embodiment provides a process for preparing a
compound of formula I:
##STR00171##
wherein Y is:
##STR00172##
and the other variables are as defined in any of the embodiments
herein;
[0120] comprising reacting a compound of formula 7:
##STR00173##
wherein the variables are as defined in any of the embodiments
herein, and a compound of formula RNHCH(R.sup.2)C(O)X, wherein X is
OH or an appropriate derivative, in the presence of conditions for
coupling an amine and an acid (when X is OH) or an appropriate acid
derivative (when X is not OH; for example, X is Cl) to provide the
compound of formula I.
[0121] Yet another embodiment of this invention provides a process
for preparing a compound of formula II:
##STR00174##
wherein the variables are as defined in any of the embodiments
herein, comprising reacting a compound of formula I:
##STR00175##
wherein Y is:
##STR00176##
wherein R and R.sup.1 are each independently as defined in any of
the embodiments herein, under hydrolysis conditions, to provide the
compound of formula II. In certain embodiments, R is
R.sup.3C(.dbd.O). In yet other embodiments, when A is proline, R is
R.sup.3C(.dbd.O). Hydrolysis conditions for converting I to II are
well known to skilled practitioners (see e.g., Greene). Such
conditions include an appropriate solvent (e.g., acetonitrile) and
aqueous acid (e.g., 2M HCl).
[0122] Another embodiment provides a process for preparing a
compound of formula 6-A:
##STR00177##
wherein PG.sub.2 is a suitable nitrogen protecting group and
R.sup.1 is as defined in any of the embodiments herein, comprising
reacting a compound of formula 5-A:
##STR00178##
under suitable ring cyclization conditions, to provide the compound
of formula 6-A. Suitable ring cyclization conditions include an
acid and a suitable solvent; for example, TFA in DCM.
[0123] Another embodiment provides a process for preparing a
compound of formula 5-A:
##STR00179##
comprising reacting a compound of formula 4-A:
##STR00180##
in the presence of R.sup.1--OH (or a suitable acetal forming
reagent), protic or Lewis acid (for example, TsOH), and a suitable
solvent to provide the compound of formula 5-A.
[0124] Another embodiment provides a process for preparing a
compound of formula 4-A:
##STR00181##
comprising reacting a compound of formula 3-A:
##STR00182##
under suitable oxidation conditions (for example, a Swern
oxidation: Mancuso, A. J.; Swern, D. Synthesis, 1981, 165-185) to
provide the compound of formula 4-A. Preferred oxidation conditions
include a TEMPO oxidation (see Example I-1, Method C, below).
[0125] Another embodiment provides a process for preparing a
compound of formula 3-A:
##STR00183##
comprising:
[0126] reacting a compound of formula 2:
##STR00184##
[0127] with a compound of formula 20-A:
##STR00185##
under conditions for coupling an amine and a carboxylic acid (when
X is OH), or an amine and an appropriate carboxylic acid (when X is
not OH), to provide the compound of formula 3-A.
[0128] Another embodiment provides a process for preparing a
compound of formula 6:
##STR00186##
wherein PG.sub.2 is a suitable nitrogen protecting group and
R.sup.1 is as defined in any of the embodiments herein, comprising
reacting a compound of formula 5:
##STR00187##
under suitable cyclization conditions, to provide the compound of
formula 6.
[0129] Another embodiment provides a process for preparing a
compound of formula 5:
##STR00188##
comprising reacting a compound of formula 4:
##STR00189##
in the presence of R.sup.1--OH (or a suitable acetal forming
reagent), protic or Lewis acid (for example, TsOH), and a suitable
solvent to provide the compound of formula 5. Preferably, the
solvent is CH.sub.2Cl.sub.2, toluene, or chlorobenzene.
[0130] Another embodiment provides a process for preparing a
compound of formula 4:
##STR00190##
comprising reacting a compound of formula 3:
##STR00191##
under suitable oxidation conditions (for example a Swern oxidation)
to provide the compound of formula 4. Preferred oxidation
conditions include a TEMPO oxidation (see Example I-1, Method C,
below).
[0131] Another embodiment provides a process for preparing a
compound of formula 3:
##STR00192##
comprising:
[0132] reacting a compound of formula 2:
##STR00193##
[0133] with a compound of formula 20:
##STR00194##
under conditions for coupling an amine and a carboxylic acid (when
X is OH), or an amine and an appropriate carboxylic acid (when X is
not OH), to provide the compound of formula 3.
[0134] Another embodiment provides a process for preparing a
compound of formula 16:
##STR00195##
wherein PG.sub.2 is a suitable nitrogen protecting group and
R.sup.1 is as defined in any of the embodiments herein, comprising
reacting a compound of formula 15:
##STR00196##
under suitable cyclization conditions, to provide the compound of
formula 16.
[0135] Another embodiment provides a process for preparing a
compound of formula 15:
##STR00197##
comprising reacting a compound of formula 14:
##STR00198##
in the presence of R.sup.1--OH (or a suitable acetal forming
reagent), protic or Lewis acid (for example, TsOH), and a suitable
solvent to provide the compound of formula 15.
[0136] Another embodiment provides a process for preparing a
compound of formula 14:
##STR00199##
comprising reacting a compound of formula 13:
##STR00200##
under suitable oxidation conditions (example, a Swern oxidation) to
provide the compound of formula 14.
[0137] Another embodiment provides a process for preparing a
compound of formula 13:
##STR00201##
comprising reacting a compound of formula 2 with a compound of
formula 21:
##STR00202##
under conditions for coupling an amine and a carboxylic acid (when
X is OH), or an amine and an appropriate carboxylic acid (when X is
not OH), to provide the compound of formula 13.
[0138] Another embodiment provides a process for preparing a
compound of formula 22:
##STR00203##
comprising reacting a compound of formula 23:
##STR00204##
in the presence of R.sup.1--OH (or a suitable acetal forming
reagent), protic or Lewis acid (for example, TsOH), and a suitable
solvent to provide the compound of formula 22. Acetal forming
equivalents include, but are not limited to, triethylorthoformate,
a diethylacetal, such as a
(CH.sub.3).sub.2C(OCH.sub.2CH.sub.3).sub.2. Preferably, the solvent
is CH.sub.2Cl.sub.2, toluene, or chlorobenzene.
[0139] Another embodiment provides a process for preparing a
compound of formula 23 comprising reacting a compound of formula
2:
##STR00205##
[0140] under suitable oxidation conditions (example Swern) to
provide the compound of formula 23.
[0141] Another embodiment provides a process for preparing a
compound of formula 5-A
##STR00206##
wherein PG.sub.1 is a suitable carboxylic acid protecting group,
PG.sub.2 is a suitable nitrogen-protecting group, and R.sup.1 is as
defined in any one of claims 1 or 5-9, comprising:
[0142] reacting a compound of formula 20-A:
##STR00207##
[0143] with a compound of formula 22
##STR00208##
under conditions for coupling an amine and a carboxylic acid (when
X is OH), or an amine and an appropriate carboxylic acid (when X is
an appropriate leaving group), to provide the compound of formula
5-A.
[0144] Another embodiment provides a process for preparing a
compound of formula 5:
##STR00209##
comprising reacting a compound of formula 20:
##STR00210##
with a compound of formula 22
##STR00211##
under conditions for coupling an amine and a carboxylic acid (when
X is OH), or an amine and an appropriate carboxylic acid (when X is
not OH), to provide the compound of formula 5.
[0145] Another embodiment provides a process for preparing a
compound of formula 5-A:
##STR00212##
comprising reacting a compound of formula 21:
##STR00213##
with a compound of formula 22
##STR00214##
under conditions for coupling an amine and a carboxylic acid (when
X is OH), or an amine and an appropriate carboxylic acid (when X is
not OH), to provide the compound of formula 5-A.
[0146] In accordance with this invention, the processes may be used
alone or in combination to provide a compound of this
invention.
[0147] Certain specific embodiments of this invention provide
processes for preparing compounds 4 from 3 (in embodiments where
compounds 4 are isolated); 5 from 3 (in embodiments where compounds
4 is not isolated but carried on directly, e.g., generated in
situ); 5 from 4; and 6 from 5 according to the methods disclosed
herein. In a preferred embodiment, compounds 6 are prepared from
compounds 5; compounds 5 are prepared from compounds 4 (whether
isolated or not); and compounds 4 are prepared from 3. Preferably,
compounds 6 are used in the preparation of proline containing
caspase inhibitors. Such proline containing caspase inhibitors
include, but are not limited to, those disclosed in WO 95/35308, WO
99/47545, WO 01/81330, and WO 01/90063 (which are all incorporated
herein by reference). For example, compound IA (and stereoisomers
thereof) of WO 01/90063 (which are specifically incorporated herein
by reference) could be prepared as disclosed herein (see, e.g.,
page 13). For the avoidance of doubt, it should be understood that
such proline containing compounds could be depicted by formula I
except that Ring A is pyrrolidine
##STR00215##
(i.e. is derived from proline).
[0148] The processes for converting compounds 6 to proline
containing caspase inhibitors are preferably as disclosed herein.
The processes for preparing compounds 3 are also preferably as
disclosed herein. However other processes known to skilled
practitioners could be used to convert compounds 6 to proline
containing caspase inhibitors and/or to prepare compounds 3.
[0149] Other embodiments of this invention provide the compounds of
formula 3 to 6, 3-A to 6-A, and 13-16.
[0150] One embodiment of this invention provides the compounds of
formula 4A:
##STR00216##
[0151] Another embodiment of this invention provides the compounds
of formula 4:
##STR00217##
[0152] Another embodiment of this invention provides the compounds
of formula 14:
##STR00218##
[0153] One embodiment of this invention provides the compounds of
formula 5-A:
##STR00219##
[0154] Another embodiment of this invention provides the compounds
of formula 5:
##STR00220##
[0155] Another embodiment of this invention provides the compounds
of formula 15:
##STR00221##
[0156] One embodiment of this invention provides the compounds of
formula 3-A:
##STR00222##
[0157] Another embodiment of this invention provides the compounds
of formula 3:
##STR00223##
[0158] Another embodiment of this invention provides the compounds
of formula 13:
##STR00224##
[0159] In all the above embodiments, the variables are as defined
in any of the embodiments herein. In a preferred form of 3,
PG.sub.2 is Z and PG.sub.1 is C.sub.1-6 straight-chained or
branched alkyl group (preferably a t-butyl group), either alone or
in combination.
[0160] As would be realized by skilled practitioners certain
process steps may be accomplished in discrete steps or in situ. For
example, deprotection and subsequent reaction of an amine may be
accomplished by step-wise (by isolating the amine) or in a one step
procedure (without isolating the amine).
[0161] In certain embodiments, the above processes are conducted as
described herein (e.g., in the schemes, examples, and accompanying
description).
[0162] Compounds such as 3 could be used in processes for preparing
proline containing compounds, such as caspase inhibitors. Proline
containing caspase inhibitors include, but are not limited to,
those disclosed in WO 95/35308, WO 99/47545, WO 01/81330, and WO
01/90063 (which are all incorporated herein by reference). For
example, compound IA (and stereoisomers thereof) of WO 01/90063
(which are specifically incorporated herein by reference) could be
prepared as disclosed herein (see, e.g., page 13).
[0163] 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.
[0164] For example, a carboxylic acid group in a compound of this
invention may be derivatized as, for example, an ester. Preferred
esters would be those derived from:
[0165] a C.sub.1-6 straight-chained or branched alkyl, alkenyl, or
alkynyl, wherein the alkyl, alkenyl, or alkynyl is optionally
substituted with C.sub.6-10aryl, CF.sub.3, Cl, F, OMe, OEt,
OCF.sub.3, CN, or NMe.sub.2;
[0166] a C.sub.1-6 cycloalkyl, wherein 1-2 carbon atoms in the
cycloalkyl is optionally replaced with --O-- or --NR.sup.9--.
[0167] Compounds of this invention having a carbonyl group may be
similarly derivatized as, e.g., an acetal, ketal, oxime
(.dbd.NOR.sup.9), hydrazine (.dbd.NN(R.sup.9).sub.2), thioacetal,
or thioketal.
[0168] Appropriate derivatives of amines are known in the art and
are also included within the scope of this invention.
[0169] Certain of the above derivatives would include the
protective groups known to skilled practitioners (see, e.g.,
Greene). As would be recognized by a skilled practitioner, these
protective groups may also be employed in the processes of this
invention.
[0170] The compounds of this invention may be assayed for their
ability to inhibit apoptosis, the release of IL-1.beta. or caspase
activity directly. Assays for each of the activities are known in
the art. However, as would be recognized by a skilled practitioner,
a prodrug compound of this invention should be active only in
assays where the prodrug moiety would be cleaved, typically in in
vivo assays.
[0171] Assays for caspase activity are described in WO
99/47545.
[0172] According to another embodiment, the present invention
provides a pharmaceutical composition comprising:
[0173] a) a compound of the invention, as defined herein, or a
pharmaceutically acceptable salt thereof; and
[0174] b) a pharmaceutically acceptable carrier, adjuvant or
vehicle.
[0175] It should be understood that compounds and pharmaceutically
acceptable salts thereof are included within this invention are. 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.
[0176] 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.
[0177] 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-polyoxypropylene-block polymers,
polyethylene glycol and wool fat.
[0178] According to a preferred embodiment, the compositions of
this invention are formulated for pharmaceutical administration to
a mammal, preferably a human being.
[0179] 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.
[0180] 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.
[0181] 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.
[0182] 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.
[0183] 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.
[0184] 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.
[0185] 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.
[0186] 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. In one embodiment, the compositions
are as formulated in, e.g., U.S. Pat. No. 6,645,994 and/or U.S.
Pat. No. 6,630,473.
[0187] 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.
[0188] The above-described compounds and 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 (e.g., bacterial
infections, preferably, eye infections), a degenerative disease, a
disease associated with cell death, an excess dietary alcohol
intake disease, a viral mediated disease, retinal disorders,
uveitis, inflammatory peritonitis, osteoarthritis, pancreatitis,
asthma, adult respiratory distress syndrome, glomerulonephritis,
rheumatoid arthritis, systemic lupus erythematosus, scleroderma,
chronic thyroiditis, Grave's disease, autoimmune gastritis,
diabetes, autoimmune hemolytic anemia, autoimmune neutropenia,
thrombocytopenia, chronic active hepatitis, myasthenia gravis,
inflammatory bowel disease, Crohn's disease, psoriasis, atopic
dermatitis, scarring, graft vs. host disease, organ transplant
rejection, organ apoptosis after burn injury, osteoporosis,
leukemias and related disorders, myelodysplastic syndrome, multiple
myeloma-related bone disorder, acute myelogenous leukemia, chronic
myelogenous leukemia, metastatic melanoma, Kaposi's sarcoma,
multiple myeloma, hemorrhagic shock, sepsis, septic shock, burns,
Shigellosis, Alzheimer's disease, Parkinson's disease, Huntington's
disease, Kennedy's disease, prion disease, cerebral ischemia,
epilepsy, myocardial ischemia, acute and chronic heart disease,
myocardial infarction, congestive heart failure, atherosclerosis,
coronary artery bypass graft, spinal muscular atrophy, amyotrophic
lateral sclerosis, multiple sclerosis, HIV-related encephalitis,
aging, alopecia, neurological damage due to stroke, ulcerative
colitis, traumatic brain injury, spinal cord injury, hepatitis-B,
hepatitis-C, hepatitis-G, yellow fever, dengue fever, Japanese
encephalitis, various forms of liver disease, renal disease,
polycystic kidney disease, H. pylori-associated gastric and
duodenal ulcer disease, HIV infection, tuberculosis, meningitis,
toxic epidermal necrolysis, pemphigus, and autoinflammatory
diseases (sometimes referred to as autoinflammatory fever
syndromes) and related syndromes such as Muckle-Wells Syndrome
(MWS), Familial Cold Urticaria (FCU), Familial Mediterranean Fever
(FMF), Chronic Infantile Neurological Cutaneous and Articular
Syndrome (CINCAS), a.k.a. Neonatal Onset Multisystem Inflammatory
Disease (NOMID), TNFR1-Associated Periodic Syndrome (TRAPS), and
Hyper-IgD periodic fever Syndrome (HIDS). The compounds and
compositions are also useful in treating complications associated
with coronary artery bypass grafts. The compounds and compositions
are also useful for decreasing IGIF (also known as IL-18) or
IFN-.gamma. production. The compounds and compositions are also
useful in immunotherapy as a cancer treatment.
[0189] The compounds and compositions may also be used in methods
for preserving cells. These methods would be useful for preserving
organs, particularly those intended for transplant, or blood
products.
[0190] The compounds of this invention are useful as dual caspase-1
and capase-8 inhibitors. Without being bound by theory, the R.sup.2
and R.sup.3 groups of the compounds of this invention appear to be
related to this surprising activity. Bridged A groups of the
compounds of this invention, such as
##STR00225##
also appear to be related to this surprising activity. As such, the
compounds and compositions of this invention are particularly
useful in treating or preventing inflammatory conditions.
[0191] According to another embodiment, the compositions of this
invention may further comprise another therapeutic agent (i.e., one
or more additional agents). Such agents include, but are not
limited to, thrombolytic agents such as tissue plasminogen
activator and streptokinase. When an additional agent is used, the
additional 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.
[0192] The amount of compound present in the compositions of this
invention 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.
[0193] Dosage levels of between about 0.01 and about 50 or about
100 mg/kg body weight per day, preferably between 0.5 and about 75
mg/kg body weight per day and most preferably between about 1 and
about 25 or about 50 mg/kg body weight per day of the active
ingredient compound are useful in a monotherapy.
[0194] Typically, a compound or composition 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.
[0195] When the compositions of this invention comprise a
combination of a compound of this invention 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 about 100%, and more preferably between about
10% to about 80% of the dosage normally administered in a
monotherapy regime.
[0196] Upon improvement of a patient's condition, a maintenance
dose of a compound, composition or combination of this invention
may be administered, if necessary.
[0197] Subsequently, the dosage or frequency of administration, or
both, may be reduced, as a function of the symptoms, to a level at
which the improved condition is retained when the symptoms have
been alleviated to the desired level, treatment should cease.
Patients may, however, require intermittent treatment on a
long-term basis upon any recurrence of disease symptoms.
[0198] As the skilled practitioner will appreciate, lower or higher
doses than those recited above may be required. It should be
understood that a specific dosage and treatment regimens 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, the severity and course of the
particular disease, the patient's disposition to the disease being
treated, and the judgment of the treating physician. The amount of
active ingredients will also depend upon the particular compound
and other therapeutic agent, if present, in the composition.
[0199] In a preferred embodiment, the invention provides a method
of treating a patient, preferably a mammal, having one of the
aforementioned diseases, comprising the step of administering to
said patient a compound or 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.
[0200] The compounds of this invention may also be incorporated
into compositions for coating implantable medical devices, such as
prostheses, artificial valves, vascular grafts, stents and
catheters. Accordingly, the present invention, in another aspect,
includes a composition for coating an implantable device comprising
a compound of the present invention and a carrier suitable for
coating said implantable device. In still another aspect, the
present invention includes an implantable device coated with a
composition comprising a compound of the present invention and a
carrier suitable for coating said implantable device.
[0201] Another aspect of the invention relates to inhibiting
caspase activity in a biological sample, which method comprises
contacting said biological sample with a compound of this invention
or a composition comprising said compound. The term "biological
sample", as used herein, includes, without limitation, cell
cultures or extracts thereof; biopsied material obtained from a
mammal or extracts thereof; and blood, saliva, urine, feces, semen,
tears, or other body fluids or extracts thereof.
[0202] Inhibition of caspase activity in a biological sample is
useful for a variety of purposes that are known to one of skill in
the art. Examples of such purposes include, but are not limited to,
blood transfusion, organ-transplantation, biological specimen
storage, and biological assays.
[0203] The compounds of this invention are useful in methods for
preserving cells, such as may be needed for an organ transplant or
for preserving blood products. Similar uses for caspase inhibitors
have been reported [Schierle et al., Nature Medicine, 5, 97
(1999)]. The method involves treating the cells or tissue to be
preserved with a solution comprising the caspase inhibitor. The
amount of caspase inhibitor needed will depend on the effectiveness
of the inhibitor for the given cell type and the length of time
required to preserve the cells from apoptotic cell death.
[0204] Without being bound by theory, applicants' cyclic acetal
compounds are believed to be prodrugs. That is, the acetal portion
is cleaved in vivo to provide a corresponding acid-aldehyde
compound. As would be recognized by a skilled practitioner,
chemical compounds may be metabolized in vivo, e.g., at a site
other than the prodrug cleavage site. Any such metabolites are
included within the scope of this invention.
[0205] 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 I-1
(S,S,S,R)-1-[(2S)-(3-Methoxy-2-methyl-benzoylamino)-3-methyl-butyryl]-pyrr-
olidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00226##
[0206] Method A
(S)-3-Amino-4-hydroxy-butyric acid tert-butyl ester
##STR00227##
[0208] A solution of (S)-benzyloxycarbonylamino-4-hydroxy-butyric
acid tert-butyl ester (prepared as described in Michel et al,
Helvetica Chimica Acta 1999, 1960)(0.94 g) in ethyl acetate (15 ml)
was hydrogenated over palladium hydroxide/carbon (20% w/w, 160 mg).
The catalyst was removed via filtration through celite.
Concentration of the filtrate in vacuo afforded the subtitle
compound as a colorless oil (486 mg, 91%); .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 1.48 (9H, s), 1.95 (3H, brs), 2.28 (1H, dd),
2.46 (1H, dd), 3.29 (1H, brm), 3.42 (1H, m), 3.60 (1H, m).
Method B
(1S)-2-((S)-2-tert-Butoxycarbonyl-1-hydroxymethyl-ethylcarbamoyl)-pyrrolid-
ine-1-carboxylic acid benzyl ester
##STR00228##
[0210] To a stirred solution of (S)-3-Amino-4-hydroxy-butyric acid
tert-butyl ester (800 mg, 4.57 mmol) and Z-Pro-OH (1.14 g, 4.57
mmol) in THF (30 ml) was added 2-hydroxybenzotriazole hydrate (741
mg, 1.2 eq,), DMAP (698 mg, 1.25 eq.), diisopropylethylamine (1.03
ml, 1.3 eq.) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride (EDC, 1.05 g, 1.2 eq.). The resulting mixture was
stirred at ambient temperature for 18 hours then diluted with ethyl
acetate. The mixture was then washed with water, saturated aqueous
sodium bicarbonate solution and brine, dried over magnesium
sulfate, filtered and concentrated under reduced pressure. The
residue was purified by flash chromatography (60% ethyl
acetate/petrol) to afford the sub-title compound as a colorless
solid (1.483 g, 90%); MS ES (+) 407.3.
Method C
(1S)-2-((S)-2-tert-Butoxycarbonyl-1-formyl-ethylcarbamoyl)-pyrrolidine-1-c-
arboxylic acid benzyl ester
##STR00229##
[0212] A solution of
(1S)-2-((S)-2-tert-Butoxycarbonyl-1-hydroxymethyl-ethylcarbamoyl)-pyrroli-
dine-1-carboxylic acid benzyl ester (10 g) in DCM (100 ml) was
cooled to 0.degree. C. under nitrogen.
2,2,6,6-tetramethylpiperidinyloxy (TEMPO, 38 mg) was then added
followed by trichloroisocyanuric acid (6 g) portionwise over 30
minutes. The mixture was stirred at ambient temperature for 2
hours, then filtered through celite. The filtrate was washed with
water, 1M sodium thiosulfate solution and water. Drying over
magnesium sulfate and concentration under reduced pressure gave the
sub-title compound as a pale yellow oil (9.92 g, 99%); .sup.1H NMR
(400 MHz, d-6 DMSO) .delta. 1.38 (9H, d), 1.79-1.86 (3H, m),
2.08-2.23 (1H, m), 2.36-2.51 (1H, 2.times.dd), 2.61-2.86 (1H,
2.times.dd), 3.88-3.46 (2H, m), 4.24-4.30 (2H, m), 5.05 (2H, quin),
7.28-7.37 (5H, m), 8.59-8.64 (1H, 2.times.d), 9.21 (0.57H, s), 9.37
(0.43H, s).
Method D
(1S)-2-((S)-1-tert-Butoxycarbonylmethyl-2,2-diethoxy-ethylcarbamoyl)-pyrro-
lidine-1-carboxylic acid benzyl ester
##STR00230##
[0214] To a solution of
(1S)-2-((S)-2-tert-Butoxycarbonyl-1-formyl-ethylcarbamoyl)-pyrrolidine-1--
carboxylic acid benzyl ester (4.98 g) in dichloromethane (70 ml)
was added triethyl orthoformate (6.2 mL) and p-toluenesulfonic acid
monohydrate (47 mg). The resulting mixture was stirred at ambient
temperature until no aldehyde remained by TLC. The mixture was
concentrated in vacuo, the re-dissolved in dichloromethane (35 mL).
Saturated aqueous sodium bicarbonate solution (35 mL) was then
added and the organic phase removed. This was washed with water and
brine, dried (magnesium sulfate), filtered and concentrated under
reduced pressure. This gave the sub-title compound as a pale yellow
oil (4.85 g, 82%); .sup.1H NMR (400 MHz, d-6 DMSO) .delta.
1.04-1.11 (6H, m), 1.35-1.37 (9H, m), 1.73-1.89 (3H, m), 2.01-2.49
(3H, m), 3.43-3.52 (6H, m), 4.05-4.29 (3H, m), 4.96-5.06 (2H, m),
7.27-7.38 (5H, m), 7.80 (0.5H, d), 7.88 (0.5H, d).
Method E
(1S)-2-((2R,3S)-2-Ethoxy-5-oxo-tetrahydro-furan-3-ylcarbamoyl)-pyrrolidine-
-1-carboxylic acid benzyl ester 6.1
(1S)-2-((2S,3S)-2-Ethoxy-5-oxo-tetrahydro-furan-3-ylcarbamoyl)-pyrrolidine-
-1-carboxylic acid benzyl ester 6.2
##STR00231##
[0216] A solution of
(1S)-2-((S)-1-tert-Butoxycarbonylmethyl-2,2-diethoxy-ethylcarbamoyl)-pyrr-
olidine-1-carboxylic acid benzyl ester (4.85 g) in dichloromethane
(25 ml) was cooled to 0.degree. C. under nitrogen. Trifluoroacetic
acid (6 ml) was then added and the mixture stirred at 0.degree. C.
for 15 minutes, then warmed to ambient temperature and stirred
until the reaction was complete by TLC. The mixture was then
diluted with dichloromethane (90 ml) and saturated aqueous sodium
bicarbonate solution (130 ml) and stirred for 15 minutes. The
organic phase was then removed and washed with 1:1 saturated
aqueous sodium bicarbonate/brine (100 ml), the combined aqueous
washings was re-extracted with DCM (100 ml) and the combined
organic layers dried (magnesium sulfate), filtered and concentrated
under reduced pressure. This afforded the sub-title compound as a
mixture of epimers at the ketal centre (C2). The epimers were
separated on silica gel, eluting with 30% acetone/petrol.
Syn-isomer 6.1 (white solid); .sup.1H NMR (400 MHz, d-6 DMSO)
.delta. 1.08-1.17 (3H, m), 1.78-2.01 (3H, m), 2.08-2.12 (1H, m),
2.37-2.57 (1H, 2.times.dd), 2.61-2.79 (1H, 2.times.dd), 3.35-3.51
(2H, m), 3.55-3.68 (1H, m), 3.71-3.82 (1H, d), 4.20-4.32 (1H, m),
4.52-4.61 (1H, m), 4.98-5.11 (2H, m), 5.53-5.58 (1H, m), 7.24-7.42
(5h, m), 8.25-8.31 (1H, m); MS ES+377.3 (100%), ES-375.3 (10%);
Anti-isomer 6.2 (colorless oil); .sup.1H NMR (400 MHz, d-6 DMSO)
.delta. 1.08-1.19 (3H, m), 1.78-1.89 (3H, m), 2.10-2.34 (1H, m),
2.92-3.07 (1H, 2.times.dd), 3.36-3.51 (3H, m), 3.62-3.78 (2H, m),
4.12-4.21 (2H, m), 4.97-5.12 (3H, m), 7.28-7.40 (5H, m), 8.51-8.58
(1H, m); MS ES+377.4 (100%), ES-375.3 (10%).
(1S)-2-((2R,3S)-2-Methoxy-5-oxo-tetrahydro-furan-3-ylcarbamoyl)-pyrrolidin-
e-1-carboxylic acid benzyl ester 6.3
(1S)-2-((2S,3S)-2-Methoxy-5-oxo-tetrahydro-furan-3-ylcarbamoyl)-pyrrolidin-
e-1-carboxylic acid benzyl ester 6.4
##STR00232##
[0218] Prepared in a similar manner to that described in methods
A-E, using trimethylorthoformate in step D, to afford the sub-title
compounds as a mixture of epimers 6.3 and 6.4. The epimers were
separated on silica gel eluting with 30% to 40% 2-Butanone/Petrol
to 70% Acetone/Petrol. Syn-isomer 6.3 (viscous colorless oil);
.sup.1H NMR (400 MHz, d-6 DMSO) .delta. 1.77-1.89 (3H, m),
2.07-2.12 (1H, m), 2.32-2.43 (1H, 2.times.d), 2.55-2.61 (1H,
2.times.d), 2.71-2.81 (1H, 2.times.d), 3.39-3.62 (4H, m), 4.21-4.30
(1H, m), 4.57-4.64 (1H, m), 5.01-5.09 (2H, m), 5.42-5.47 (1H, m),
7.27-7.42 (5H, m), 8.24-8.31 (1H, m); Anti-isomer 6.4 (white
solid); .sup.1H NMR (400 MHz, d-6 DMSO) .delta. 1.79-1.90 (3H, m),
2.09-2.21 (1H, m), 2.23-41 (1H, 2.times.d), 2.91-3.05 (1H,
2.times.dd), 3.35-3.71 (5H, m), 4.09-4.21 (2H, m), 4.98-5.19 (3H,
m), 7.28-7.41 (5H, m), 8.51-8.58 (1H, m).
(1S)-2-((2R,3S)-2-Isopropoxy-5-oxo-tetrahydro-furan-3-ylcarbamoyl)-pyrroli-
dine-1-carboxylic acid benzyl ester 6.5
(1S)-2-((2S,3S)-2-Isopropoxy-5-oxo-tetrahydro-furan-3-ylcarbamoyl)-pyrroli-
dine-1-carboxylic acid benzyl ester 6.6
##STR00233##
[0220] Prepared in a similar manner to that described in Methods
A-E, using triisopropylorthoformate in step D, to afford the
sub-title compound as a mixture of epimers 6.5 and 6.6. The epimers
were separated on silica gel eluting with 30% to 40%
2-Butanone/Petrol. Syn-isomer 6.5 (colorless gum); .sup.1H NMR (400
MHz, d-6 DMSO) .delta. 1.07-1.16 (6H, m), 1.81-1.86 (2H, m),
2.37-2.71 (2H, m), 3.35-3.53 (2H, m), 3.86-3.90 (1H, m), 4.18-4.24
(1H, m), 4.46-4.55 (1H, m), 4.95-5.10 (2H, m), 5.63 (1H, d),
7.27-7.38 (5H, m), 8.22-8.30 (1H, m); MS ES+391.3 (100%);
Anti-isomer 6.6 (white solid); .sup.1H NMR (400 MHz, d-6 DMSO)
.delta. 1.07-1.15 (6H, m), 1.78-1.82 (3H, m), 2.07-2.41 (2H, m),
2.87-3.01 (1H, m), 3.35-3.50 (2H, m), 3.74-3.96 (1H, m), 4.07-4.18
(2H, m), 4.95-5.11 (2H, m), 5.22 (1H, 2.times.s), 7.24-7.39 (5H,
m), 8.48-8.53 (1H, m); MS ES+391.4 (100%).
(1S)-2-((2R,3S)-2-Propoxy-5-oxo-tetrahydro-furan-3-ylcarbamoyl)-pyrrolidin-
e-1-carboxylic acid benzyl ester 6.7
(1S)-2-((2S,3S)-2-Propoxy-5-oxo-tetrahydro-furan-3-ylcarbamoyl)-pyrrolidin-
e-1-carboxylic acid benzyl ester 6.8
##STR00234##
[0222] Prepared in a similar manner to that described in methods
A-E, using tripropylorthoformate in step D, to afford the sub-title
compounds as a mixture of epimers 6.7 and 6.8. The epimers were
separated on silica gel eluting with 30% to 40% 2-Butanone/Petrol.
Syn-isomer 6.7 (colorless gum); .sup.1H NMR (400 MHz, d-6 DMSO)
.delta. 0.84-0.93 (3H, m), 1.55 (2H, m), 1.81-1.89 (3H, m),
2.08-2.22 (1H, m), 2.37-2.61 (1H, 2.times.dd), 2.71-2.80 (1H,
2.times.dd), 3.31-3.53 (2H, m), 3.60-3.69 (1H, m), 4.20-4.29 (1H,
m), 4.52-4.61 (1H, m), 4.95-5.11 (2H, m), 5.50 (1H, m), 7.27-7.36
(5H, m), 8.27 (1H, m); Anti-isomer 6.8 (colorless oil); .sup.1H NMR
(400 MHz, d-6 DMSO) .delta. 0.82-0.90 (3H, m), 1.46-1.57 (2H, m),
1.77-1.89 (3H, m), 2.06-2.41 (1H, m), 2.90-3.05 (1H, 2.times.dd),
3.33-3.66 (5H, m), 4.11-4.20 (2H, m), 4.94-5.10 (3H, m), 7.28-7.37
(5H, m), 8.51 (1H, m).
(1S)-2-((2R,3S)-2-Butoxy-5-oxo-tetrahydro-furan-3-ylcarbamoyl)-pyrrolidine-
-1-carboxylic acid benzyl ester 6.9
(1S)-2-((2S,3S)-2-Butoxy-5-oxo-tetrahydro-furan-3-ylcarbamoyl)-pyrrolidine-
-1-carboxylic acid benzyl ester 6.10
##STR00235##
[0224] Prepared in a similar manner to that described in methods
A-E, using tributylorthoformate in step D, to afford the sub-title
compounds as a mixture of epimers 6.9 and 6.10. The epimers were
separated on silica gel eluting with 30% to 40% 2-Butanone/Petrol.
Syn-isomer 6.9 (colorless gum); .sup.1H NMR (400 MHz, d-6 DMSO)
.delta. 0.86-0.92 (3H, m), 1.28-1.37 (2H, m), 1.45-1.54 (2H, m),
1.79-1.88 (3H, m), 2.07-2.21 (1H, m), 2.35-2.78 (2H, m), 3.31-3.54
(2H, m), 3.63-3.70 (1H, m), 4.21-4.29 (1H, m), 4.51-4.61 (1H, m),
4.95-5.09 (2H, m), 5.50 (1H, m), 7.27-7.37 (5H, m), 8.25 (1H, m);
Anti-isomer 6.10 (colorless oil); .sup.1H NMR (400 MHz, d-6 DMSO)
.delta. 0.85-0.93 (3H, m), 1.26-1.36 (2H, m), 1.44-1.56 (2H, m),
1.77-1.90 (3H, m), 2.08-2.40 (1H, m), 2.89-3.05 (1H, 2.times.dd),
3.34-3.70 (5H, m), 4.08-4.19 (2H, m), 4.95-5.10 (3H, m), 7.28-7.39
(5H, m), 8.53 (1H, m).
Method F
{(S)-1-[(1R,3S,4S)-3-((2R,3S)-2-Ethoxy-5-oxo-tetrahydro-furan-3-ylcarbamoy-
l)-2-pyrrolidine-2-carbonyl]-2,2-dimethyl-propyl}-carbamic acid
benzyl ester
##STR00236##
[0226] To a solution of
(1S)-2-((2R,3S)-2-Ethoxy-5-oxo-tetrahydro-furan-3-ylcarbamoyl)-pyrrolidin-
e-1-carboxylic acid benzyl ester 6.1 (4.68 g) in ethyl acetate (160
ml) and DMF (25 ml) was added triethylamine (2.5 g) followed by
palladium hydroxide/carbon (20% w/w, 1 g). The mixture was stirred
under an atmosphere of hydrogen until no starting material was
present by TLC. The catalyst was removed by filtration through
celite. To the filtrate was added
(S)-2-benzyloxycarbonylamino-3,3-dimethyl-butyric acid (4.93 g),
hydroxybenzotriazole hydrate (2.01 g) and
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC,
2.85 g). The resulting mixture was stirred at ambient temperature
overnight. Saturated aqueous sodium bicarbonate solution (180 ml)
was then added and the organic phase removed. This was washed with
saturated aqueous ammonium chloride (180 ml), then brine (180 ml),
dried (magnesium sulfate), filtered and concentrated under reduced
pressure. The crude product was purified on silica gel, eluting
with 40-75% ethyl acetate/petrol. The sub-title compound was
obtained as a white foam (4.02 g, 66%); .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 0.97 (9H, s), 1.14 (3H, t), 1.79-1.94 (3H, m),
2.02-2.10 (1H, m), 2.44 (1H, dd), 2.75 (1H, dd), 3.52-3.66 (2H, m),
3.70-3.79 (2H, m), 4.22 (1H, d), 4.38-4.41 (1H, m), 4.48-4.58 (1H,
m), 5.03 (2H, q), 5.56 (1H, d), 7.26 (1H, d), 7.29-7.40 (5H, m),
8.24 (1H, d); MS ES+490.6 (100%), ES-488.8 (10%).
Method G
(S,S,S,R)-1-[(2S)-(3-Methoxy-2-methyl-benzoylamino)-3-methyl-butyryl]-pyrr-
olidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00237##
[0228] To a solution of
{(S)-1-[(1R,3S,4S)-3-((2R,3S)-2-Ethoxy-5-oxo-tetrahydro-furan-3-ylcarbamo-
yl)-2-pyrrolidine-2-carbonyl]-2,2-dimethyl-propyl}-carbamic acid
benzyl ester (344 mg) in ethyl acetate (20 ml)was added palladium
hydroxide/carbon (20% w/w, 74 mg). The mixture was stirred under an
atmosphere of hydrogen until no starting material was present by
TLC. The catalyst was removed by filtration through celite and the
filtrate concentrated under reduced pressure to give the amine as a
brown foam (260 mg). A portion of this material (153 mg) was
dissolved in THF and 3-methoxy-2-methyl benzoic acid (146 mg),
diisopropylamine (191 .mu.l), hydroxybenzotriazole hydrate (77 mg)
and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride
(EDC, 109 mg) were added. The resulting mixture was stirred at
ambient temperature for 24 hours then diluted with saturated
aqueous sodium bicarbonate. The organic phase was removed and
washed with saturated aqueous ammonium chloride, then brine, dried
(magnesium sulfate), filtered and concentrated under reduced
pressure. The crude product was purified on silica gel, eluting
with ethyl acetate. This gave the sub-title compound as a white
solid (138 mg, 62%); analytical data summarized in Table 3.
[0229] Compounds of formula I-2 to I-58 have been prepared by
methods substantially similar to those described in Example
I-1.
Example I-2
(S,S,S,R)-1-[(2S)-(2-Methoxy-benzoylamino)-3-methyl-butyryl]-pyrrolidine-(-
2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00238##
[0230] Example I-3
(S,S,S,R)-1-[3-Methyl-(2S)-(2-trifluoromethoxy-benzoylamino)-butyryl]-pyrr-
olidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00239##
[0231] Example I-4
(S,S,S,R)-1-[(2S)-(3-Hydroxy-2-methyl-benzoylamino)-3-methyl-butyryl]-pyrr-
olidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00240##
[0232] Example I-5
(S,S,S,R)-1-[(2S)-(3-Amino-2-methyl-benzoylamino)-3-methyl-butyryl]-pyrrol-
idine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00241##
[0233] Example I-6
(S,S,S,R)-1-[(2S)-(2,6-Dichloro-benzoylamino)-3-methyl-butyryl]-pyrrolidin-
e-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00242##
[0234] Example I-7
(S,S,S,R)--N-{(1S)-[(2S)-((2R)-Ethoxy-5-oxo-tetrahydro-furan-(3S)-ylcarbam-
oyl)-pyrrolidine-1-carbonyl]-2-methyl-propyl}-2-methyl-nicotinamide
##STR00243##
[0235] Example I-8
(S,S,S,R)--N-{(1S)-[(2S)-((2R)-Ethoxy-5-oxo-tetrahydro-furan-(3S)-ylcarbam-
oyl)-pyrrolidine-1-carbonyl]-2-methyl-propyl}-4-methyl-nicotinamide
##STR00244##
[0236] Example I-9
(S,S,S,R)-1-{3-Methyl-(2S)-[(3-methyl-thiophene-2-carbonyl)-amino]-butyryl-
}-pyrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00245##
[0237] Example I-10
(S,S,S,R)-2,3-Dichloro-N-{(1S)-[(2S)-((2R)-ethoxy-5-oxo-tetrahydro-furan-(-
3S)-ylcarbamoyl)-pyrrolidine-1-carbonyl]-2-methyl-propyl}-isonicotinamide
##STR00246##
[0238] Example I-11
(S,S,S,R)-3,5-Dichloro-N-{(1S)-[(2S)-((2R)-ethoxy-5-oxo-tetrahydro-furan-(-
3S)-ylcarbamoyl)-pyrrolidine-1-carbonyl]-2-methyl-propyl}-isonicotinamide
##STR00247##
[0239] Example I-12
(S,S,S,R)-1-[(2S)-(3-Methoxy-2-methyl-benzoylamino)-3,3-dimethyl-butyryl]--
pyrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00248##
[0240] Example I-13
(S,S,S,R)-1-[(2S)-(3-Methoxy-2-methyl-benzoylamino)-3-methyl-butyryl]-pyrr-
olidine-(2S)-carboxylic acid
[(2R)-methoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00249##
[0241] Example I-14
(S,S,S,R)-1-[(2S)-(3-Methoxy-2-methyl-benzoylamino)-3-methyl-butyryl]-pyrr-
olidine-(2S)-carboxylic acid
[(2R)-isopropoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00250##
[0242] Example I-15
(S,S,S,R)-1-[(2S)-(3-Methoxy-2-methyl-benzoylamino)-3-methyl-butyryl]-pyrr-
olidine-(2S)-carboxylic acid
[5-oxo-(2R)-propoxy-5-tetrahydro-furan-(3S)-yl]-amide
##STR00251##
[0243] Example I-16
(S,S,S,R)-1-[(2S)-(2-Chloro-benzoylamino)-3-methyl-butyryl]-pyrrolidine-(2-
S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00252##
[0244] Example I-17
(S,S,S,R)-1-[3,3-Dimethyl-(2S)-(2-methyl-benzoylamino)-butyryl]-pyrrolidin-
e-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00253##
[0245] Example I-18
(S,S,S,R)-1-[3-Methyl-2
(S)-(2-trifluoromethyl-benzoylamino)-butyryl]-pyrrolidine-(2S)-carboxylic
acid [(2R)-ethoxy-5-oxo-tetrahydro-furan-3(S)-yl]-amide
##STR00254##
[0246] Example I-19
(S,S,S,R)-1-[2
(S)-(2-Chloro-benzoylamino)-3,3-dimethyl-butyryl]-pyrrolidine-(2S)-carbox-
ylic acid [(2R)-methoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00255##
[0247] Example I-20
(S,S,S,R)-1-[3,3-Dimethyl-(2S)-(2-trifluoromethyl-benzoylamino)-butyryl]-p-
yrrolidine-(2S)-carboxylic acid
[(2R)-isopropoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00256##
[0248] Example I-21
(S,S,S,R)-1-[(2S)-(2-Chloro-benzoylamino)-3,3-dimethyl-butyryl]-pyrrolidin-
e-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00257##
[0249] Example I-22
(S,S,S,R)-1-[3,3-Dimethyl-(2S)-(2-trifluoromethyl-benzoylamino)-butyryl]-p-
yrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00258##
[0250] Example I-23
(S,S,S,R)-1-[(2S)-(2-Chloro-benzoylamino)-3,3-dimethyl-butyryl]-pyrrolidin-
e-(2S)-carboxylic acid
[5-oxo-(2R)-propoxy-tetrahydro-furan-(3S)-yl]-amide
##STR00259##
[0251] Example I-24
(S,S,S,R)-1-[(2S)-(2-Chloro-benzoylamino)-3,3-dimethyl-butyryl]-pyrrolidin-
e-(2S)-carboxylic acid
[(2R)-butoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00260##
[0252] Example I-25
(S,S,S,R)-1-[(2S)-(2-Chloro-3-trifluoromethoxy-benzoylamino)-3,3-dimethyl--
butyryl]-pyrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00261##
[0253] Example I-26
(S,S,S,R)-1-[(2S)-(2-Chloro-benzoylamino)-3-methyl-butyryl]-pyrrolidine-(2-
S)-carboxylic acid
[5-oxo-(2R)-propoxy-tetrahydro-furan-(3S)-yl]-amide
##STR00262##
[0254] Example I-27
(S,S,S,S)-1-[(2S)-(2-Chloro-benzoylamino)-3-methyl-butyryl]-pyrrolidine-(2-
S)-carboxylic acid
[5-oxo-(2S)-propoxy-tetrahydro-furan-(3S)-yl]-amide
##STR00263##
[0255] Example I-28
(S,S,S,S)-1-[(2S)-(2-Chloro-benzoylamino)-3-methyl-butyryl]-pyrrolidine-(2-
S)-carboxylic acid
[(2S)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00264##
[0256] Example I-29
(S,S,S,R)-1-[(2S)-(2-Chloro-benzoylamino)-3-methyl-butyryl]-pyrrolidine-(2-
S)-carboxylic acid
[(2R)-butoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00265##
[0257] Example I-30
(S,S,S,S)-1-[(2S)-(2-Chloro-benzoylamino)-3-methyl-butyryl]-pyrrolidine-(2-
S)-carboxylic acid
[(2S)-butoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00266##
[0258] Example I-31
(S,S,S,R)-1-[(2S)-(2-Chloro-benzoylamino)-3-methyl-butyryl]-pyrrolidine-(2-
S)-carboxylic acid
[(2R)-isopropoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00267##
[0259] Example I-32
(S,S,S,S)-1-[(2S)-(2-Chloro-benzoylamino)-3-methyl-butyryl]-pyrrolidine-(2-
S)-carboxylic acid
[(2S)-isopropoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00268##
[0260] Example I-33
(S,S,S,R)-1-[(2S)-(2-Chloro-3-cyclopropyloxy-benzoylamino)-3,3-dimethyl-bu-
tyryl]-pyrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00269##
[0261] Example I-34
(S,S,S,R)-1-[(2S)-(2-Chloro-3-methyl-benzoylamino)-3,3-dimethyl-butyryl]-p-
yrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00270##
[0262] Example I-35
(S,S,S,R)-1-[(2S)-(2-chloro-3-methoxy-benzoylamino)-3-methyl-butyryl]-pyrr-
olidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00271##
[0263] Example I-36
(S,S,S,R)-1-[(2S)-(2-Chloro-3-ethyl-benzoylamino)-3,3-dimethyl-butyryl]-py-
rrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00272##
[0264] Example I-37
(S,S,S,R)-1-[(2S)-(2-chloro-4-methoxy-benzoylamino)-3-methyl-butyryl]-pyrr-
olidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00273##
[0265] Example I-38
(S,S,S,R)-1-[(2S)-(2-Chloro-3-cyclopropylmethyl-benzoylamino)-3,3-dimethyl-
-butyryl]-pyrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00274##
[0266] Example I-39
(S,S,S,R)-1-[(2S)-(2-Chloro-3-hydroxy-benzoylamino)-3,3-dimethyl-butyryl]--
pyrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00275##
[0267] Example I-40
(S,S,S,R)-1-[(2S)-(2-Chloro-4-acetamido-benzoylamino)-3-methyl-butyryl]-py-
rrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00276##
[0268] Example I-41
(S,S,S,R)-1-[(2S)-(2-Chloro-3-acetamido-benzoylamino)-3,3-dimethyl-butyryl-
]-pyrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00277##
[0269] Example I-42
(S,S,S,R)-1-[(2S)-(2-methyl-3-acetamido-benzoylamino)-3,3-dimethyl-butyryl-
]-pyrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00278##
[0270] Example I-43
(S,S,S,R)-1-[(2S)-(2-Chloro-4-acetamido-benzoylamino)-3,3-dimethyl-butyryl-
]-pyrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00279##
[0271] Example I-44
(S,S,S,R)-1-[(2S)-(2-fluoro-4-acetamido-benzoylamino)-3,3-dimethyl-butyryl-
]-pyrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00280##
[0272] Example I-45
(S,S,S,R)-1-[(2S)-(2-fluoro-4-acetamido-benzoylamino)-3-methyl-butyryl]-py-
rrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00281##
[0273] Example I-46
(S,S,S,R)-1-[(2S)-(2-chloro-4-isopropyloxy-benzoylamino)-3-methyl-butyryl]-
-pyrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00282##
[0274] Example I-47
(S,S,S,R)-1-[(2S)-(2-chloro-4-hydroxy-benzoylamino)-3,3-dimethyl-butyryl]--
pyrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00283##
[0275] Example I-48
(S,S,S,R)-1-[(2S)-(2-chloro-4-methoxymethyl-benzoylamino)-3,3-dimethyl-but-
yryl]-pyrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00284##
[0276] Example I-49
(S,S,S,R)-1-[(2S)-(2-Chloro-4-isobutyrylamino-benzoylamino)-3,3-dimethyl-b-
utyryl]-pyrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00285##
[0277] Example I-50
(S,S,S,R)-1-[(2S)-(2-Chloro-4-acetamido-benzoylamino)-3-cyclohexyl]-pyrrol-
idine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00286##
[0278] Example I-51
(S,S,S,R)-1-[(2S)-(2-Chloro-4-methoxycarbonylamino-benzoylamino)-3,3-dimet-
hyl-butyryl]-pyrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00287##
[0279] Example I-52
(S,S,S,R)-1-[(2S)-(2-Chloro-3-phenoxy-benzoylamino)-3,3-dimethyl-butyryl]--
pyrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00288##
[0280] Example I-53
(S,S,S,R)-1-[(2S)-(2-Chloro-4-thiazolylamino-benzoylamino)-3,3-dimethyl-bu-
tyryl]-pyrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00289##
[0281] Example I-54
(S,S,S,R)-1-[(2S)-(3-Amino-2-chloro-benzoylamino)-3-methyl-butyryl]-pyrrol-
idine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00290##
[0282] Example I-55
(S,S,S,R)-1-[(2S)-(2-Chloro-benzoylamino)-3-thiazol-4-yl-propionyl]-pyrrol-
idine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00291##
[0283] Example I-56
(S,S,S,R)-1-[(2S)-(3-Methoxy-2-methyl-benzoylamino)-3-thiazol-4-yl-propion-
yl]-pyrrolidine-(2S)-carboxylic acid
[0284] [(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00292##
Example I-57
(S,S,S,R)-1-[(2S)-(2-Chloro-3-methoxy-benzoylamino)-3,3-dimethyl-butyryl]--
pyrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00293##
[0285] Example I-58
(S,S,S,R)-1-[(2S)-(2-Chloro-benzoylamino)-3,3-dimethyl-butyryl]-piperidine-
-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00294##
[0286] Example I-59
2-[(2S)-(3-Methoxy-2-methyl-benzoylamino)-3,3-dimethyl-butyryl]-2-(1S,4R)--
aza-bicyclo[2.2.1]heptane-(3S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00295##
[0287] Method H
(1R,3S,4S)-3((S)-2-tert-Butoxycarbonyl-1-hydroxymethyl-ethylcarbamoyl)-2-a-
za-bicyclo[2.2.1]heptane-2-carboxylic acid benzyl ester
##STR00296##
[0289] To a stirred solution of (S)-3-Amino-4-hydroxy-butyric acid
tert-butyl ester (486 mg) and
(1R,3S,4S)-2-Aza-bicyclo[2.2.1]heptane-2,3-dicarboxylic acid 2
benzyl ester (prepared as described in Tararov et al, Tett. Asymm.
2002, 13, 25-28) (767 mg) in THF (18 ml) was added
2-hydroxybenzotriazole hydrate (452 mg), DMAP (426 mg),
diisopropylethylamine (631 l) and
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC,
641 mg). The resulting mixture was stirred at ambient temperature
for 18 hours then diluted with ethyl acetate. The mixture was then
washed with water, saturated aqueous sodium bicarbonate solution
and brine, dried over magnesium sulfate, filtered and concentrated
under reduced pressure. The residue was purified by flash
chromatography (60% ethyl acetate/petrol) to afford the sub-title
compound as a colorless oil (1.1 g, 91%); .sup.1H NMR (400 MHz, d-6
DMSO) .delta. 1.13-1.25 (1H, m), 1.30-1.48 (9H, m), 1.49-1.88 (6H,
m), 2.20-2.52 (2H, m), 3.09-3.34 (2H, m), 3.64 (1H, d), 4.00-4.16
(2H, brm), 4.80 (1H, m), 4.90-5.15 (2H, m), 7.21-7.41 (5H, m),
7.50-7.75 (1H, m); MS ES (+) 433.37.
Method I
(1R,3S,4S)-3-((S)-2-tert-Butoxycarbonyl-1-formyl-ethylcarbamoyl)-2-aza-bic-
yclo[2.2.1]heptane-2-carboxylic acid benzyl ester
##STR00297##
[0291] A solution of
(1R,3S,4S)-3((S)-2-tert-Butoxycarbonyl-1-hydroxymethyl-ethylcarbamoyl)-2--
aza-bicyclo[2.2.1]heptane-2-carboxylic acid benzyl ester (1.1 g) in
DCM (10 ml) was cooled to 0.degree. C. under nitrogen.
2,2,6,6-tetramethylpiperidinyloxy (TEMPO, 4 mg) was then added
followed by trichloroisocyanuric acid (621 mg) portionwise over 30
minutes. The mixture was stirred at ambient temperature for 1 hour,
then filtered through celite. The filtrate was washed with water,
1M sodium thiosulfate solution and brine. Drying over magnesium
sulfate and concentration under reduced pressure gave the sub-title
compound as a colorless oil (698 mg, 64%); .sup.1H NMR (400 MHz,
d-6 DMSO) .delta. 1.16-1.89 (16H, m), 2.30-2.80 (2H, m), 3.68-3.81
(1H, m), 4.19 (1H, brm), 4.39 (1H, m), 4.91-5.16 (2H, m), 7.21-7.43
(5H, m), 8.45 (0.4H, d), 8.60 (0.6, d), 9.19 (0.6H, s), 9.37 (0.4H,
s).
Method J
(1R,3S,4S)-3-((S)-1-tert-Butoxycarbonylmethyl-2,2-diethoxy-ethylcarbamoyl)-
-2-aza-bicyclo[2.2.1]heptane-2-carboxylic acid benzyl ester
##STR00298##
[0293] To a solution of
(1R,3S,4S)-3-((S)-2-tert-Butoxycarbonyl-1-formyl-ethylcarbamoyl)-2-aza-bi-
cyclo[2.2.1]heptane-2-carboxylic acid benzyl ester (698 mg) in
dichloromethane (10 ml) was added triethyl orthoformate (720 mg)
and p-toluenesulfonic acid monohydrate (6 mg). The resulting
mixture was stirred at ambient temperature until no aldehyde
remained by TLC. Saturated aqueous sodium bicarbonate solution was
then added and the organic phase removed. This was washed with
water and brine, dried (magnesium sulfate), filtered and
concentrated under reduced pressure. This gave the sub-title
compound as a pale yellow oil (635 mg, 78%); .sup.1H NMR (400 MHz,
d-6 DMSO) .delta. 0.96-1.15 (6H, m), 1.26-1.84 (16H, m), 2.20-2.50
(2H, m), 3.40-3.81 (5H, m), 4.10-4.28 (2H, m), 4.37 (1H, m),
4.88-5.14 (2H, m), 7.20-7.40 (5H, m), 7.65 (0.5H, d), 7.80 (0.5H,
d).
Method K
(1R,3S,4S)-3-((2R,3S)-2-Ethoxy-5-oxo-tetrahydro-furan-3-ylcarbamoyl)-2-aza-
-bicyclo[2.2.1]heptane-2-carboxylic acid benzyl ester
##STR00299##
[0295] A solution of
(1R,3S,4S)-3-((S)-1-tert-Butoxycarbonylmethyl-2,2-diethoxy-ethylcarbamoyl-
)-2-aza-bicyclo[2.2.1]heptane-2-carboxylic acid benzyl ester (635
mg) in dichloromethane (3 ml) was cooled to 0.degree. C. under
nitrogen. Trifluoroacetic acid (0.7 ml) was then added and the
mixture stirred at 0.degree. C. for 15 minutes, then warmed to
ambient temperature and stirred until the reaction was complete by
TLC. The mixture was then diluted with dichloromethane (10 ml) and
saturated aqueous sodium bicarbonate solution (14 ml). The organic
phase was then removed and washed with 1:1 saturated aqueous sodium
bicarbonate/brine (8 ml), dried (magnesium sulfate), filtered and
concentrated under reduced pressure. This afforded the sub-title
compound as a mixture of epimers at the ketal centre. The epimers
were separated on silica gel, eluting with 30% 2-butanone/petrol.
Syn-isomer (oil) (115 mg, 23%); .sup.1H NMR (400 MHz, d-6 DMSO)
.delta. 0.80-1.91 (10H, m), 2.35-2.79 (2H, m), 3.56 (1H, m),
3.66-3.80 (2H, m), 4.18 (1H, m), 4.59 (1H, m), 4.94-5.11 (2H, m),
5.53 (1H, d), 7.20-7.40 (5H, m), 8.18 (0.5H, d), 8.27 (0.5H, d); MS
ES+403.31 (100%), ES-401.37 (15%); Anti-isomer (oil) (103 mg, 20%);
.sup.1H NMR (400 MHz, d-6 DMSO) .delta. 0.80-1.85 (10H, m),
2.25-2.60 (1H, m), 2.95 (1H, m), 3.42 (1H, m), 3.5-3.75 (2H, m),
4.88-5.15 (3H, m), 7.21-7.40 (5H, m), 8.50 (0.4H, d), 8.59 (0.6H,
d).
Method L
{(S)-1-[(1R,3S,4S)-3-((2R,3S)-2-Ethoxy-5-oxo-tetrahydro-furan-3-ylcarbamoy-
l)-2-aza-bicyclo[2.2.1]heptane-2-carbonyl]-2,2-dimethyl-propyl}-carbamic
acid benzyl ester
##STR00300##
[0297] To a solution of
(1R,3S,4S)-3-((2R,3S)-2-Ethoxy-5-oxo-tetrahydro-furan-3-ylcarbamoyl)-2-az-
a-bicyclo[2.2.1]heptane-2-carboxylic acid benzyl ester (5 g) in
ethyl acetate (160 ml) and DMF (25 ml) was added triethylamine (2.5
g) followed by palladium hydroxide/carbon (20% w/w, 1 g). The
mixture was stirred under an atmosphere of hydrogen until no
starting material was present by TLC. The catalyst was removed by
filtration through celite. To the filtrate was added
(S)-2-benzyloxycarbonylamino-3,3-dimethyl-butyric acid (4.93 g),
hydroxybenzotriazole hydrate (2.01 g) and
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC,
2.85 g). The resulting mixture was stirred at ambient temperature
overnight. Saturated aqueous sodium bicarbonate solution (180 ml)
was then added and the organic phase removed. This was washed with
saturated aqueous ammonium chloride (180 ml), then brine (180 ml),
dried (magnesium sulfate), filtered and concentrated under reduced
pressure. The crude product was purified on silica gel, eluting
with 40-75% ethyl acetate/petrol. The sub-title compound was
obtained as a white foam (5.25 g, 81%); .sup.1H NMR (400 MHz, d-6
DMSO) .delta. 0.85-1.03 (10H, m), 1.07-1.20 (3H, t), 1.30 (1H, m),
1.40 (1H, m), 1.50-1.80 (3H, m), 1.93 (1H, m), 2.40-2.50 (1H, m),
2.78 (1H, m), 3.60 (1H, m), 3.78 (1H, m), 3.89 (1H, s), 4.26 (1H,
d), 4.52 (2H, m), 4.96-5.12 (2H, m), 5.56 (1H, d), 7.10 (1H, d),
7.24-7.40 (5H, m), 8.27 (1H, d); MS ES+516.93 (100%), ES-515.05
(100%).
Method M
(1R,3S,4S)-2-[(S)-2-(3-methoxy-2-methylbenzoylamino)-3,3-dimethyl-butyryl]-
-2-aza-bicyclo[2.2.1]heptane-3-carboxylic acid
((2R,3S)-2-ethoxy-5-oxo-tetrahydro-furan-3-yl)-amide
##STR00301##
[0299] To a solution of
{(S)-1-[(1R,3S,4S)-3-((2R,3S)-2-Ethoxy-5-oxo-tetrahydro-furan-3-ylcarbamo-
yl)-2-aza-bicyclo[2.2.1]heptane-2-carbonyl]-2,2-dimethyl-propyl}-carbamic
acid benzyl ester (370 mg) in ethyl acetate (20 ml)was added
palladium hydroxide/carbon (20% w/w, 74 mg). The mixture was
stirred under an atmosphere of hydrogen until no starting material
was present by TLC. The catalyst was removed by filtration through
celite and the filtrate concentrated under reduced pressure to give
the amine as a brown foam (272 mg). A portion of this material (167
mg) was dissolved in THF and 3-methoxy-2-methyl benzoic acid (146
mg), diisopropylamine (191 l), hydroxybenzotriazole hydrate (77 mg)
and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride
(EDC, 109 mg) were added. The resulting mixture was stirred at
ambient temperature for 24 hours then diluted with saturated
aqueous sodium bicarbonate. The organic phase was removed and
washed with saturated aqueous ammonium chloride, then brine, dried
(magnesium sulfate), filtered and concentrated under reduced
pressure. The crude product was purified on silica gel, eluting
with ethyl acetate. This gave the sub-title compound as a white
solid (121 mg, 52%); .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 1.10
(9H, s), 1.28 (3H, t), 1.43-1.56 (1H, m), 1.79-1.86 (3H, m), 1.99
(1H, brd), 2.29 (3H, s), 2.30-2.37 (1H, m), 2.83 (1H, dd), 3.02
(1H, brs), 3.66-3.74 (1H, m), 3.87 (3H, s), 3.88-3.94 (1H, m), 4.16
(1H, brs), 4.54 (1H, brs), 4.66-4.74 (1H, m), 4.97 (1H, d), 5.46
(1H, d), 6.44 (1H, brd), 6.93 (1H, d), 7.00 (1H, d), 7.22 (1H, t),
7.78 (1H, brd); IR (solid) cm.sup.-1 2960, 1791, 1624, 1505, 1438,
1261, 1115, 975; MS ES+530; ES-528.
[0300] Compounds of formula 1-60 to 1-73 have been prepared by
methods substantially similar to those described in Example
1-59.
Example I-60
2-[(2S)-(2-Chloro-benzoylamino)-3,3-dimethyl-butyryl]-2-(1S,4R)-aza-bicycl-
o[2.2.1]heptane-(35)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00302##
[0301] Example I-61
2-[(2S)-(4-Acetylamino-2-chloro-benzoylamino)-3,3-dimethyl-butyryl]-2-(1S,-
4R)-aza-bicyclo[2.2.1]heptane-(35)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(35)-yl]-amide
##STR00303##
[0302] Example I-62
2-[(2S)-(2-Chloro-4-propionylamino-benzoylamino)-3,3-dimethyl-butyryl]-2-(-
1S,4R)-aza-bicyclo[2.2.1]heptane-(3S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00304##
[0303] Example I-63
2-[(2S)-(2-Chloro-3-isobutyrylamino-benzoylamino)-3,3-dimethyl-butyryl]-2--
(1S,4R)-aza-bicyclo[2.2.1]heptane-(3S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00305##
[0304] Example I-64
2-[(2S)-(2-Fluoro-3-methoxy-benzoylamino)-3,3-dimethyl-butyryl]-2-(1S,4R)--
aza-bicyclo[2.2.1]heptane-(3S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00306##
[0305] Example I-65
2-[(2S)-(2-Fluoro-3-methoxy-benzoylamino)-3-methyl-butyryl]-2-(1S,4R)-aza--
bicyclo[2.2.1]heptane-(3S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00307##
[0306] Example I-66
2-[(2S)-(3-methoxy-2-methyl-benzoylamino)-3,3-dimethyl-butyryl]-2-(1S,4R)--
aza-bicyclo[2.2.1]heptane-(3S)-carboxylic acid
[(2S)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00308##
[0307] Example I-67
2-[(2S)-(2-Chloro-benzoylamino)-3,3-dimethyl-butyryl]-2-(1S,4R)-aza-bicycl-
o[2.2.1]heptane-(3S)-carboxylic acid
[(2S)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00309##
[0308] Example I-70
2-[(2S)-(4-Acetylamino-3-chloro-benzoylamino)-3,3-dimethyl-butyryl]-2-(1S,-
4R)-aza-bicyclo[2.2.1]heptane-(3S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00310##
[0309] Example I-69
2-[(2S)-(3-Chloro-4-propionylamino-benzoylamino)-3,3-dimethyl-butyryl]-2-(-
1S,4R)-aza-bicyclo[2.2.1]heptane-(3S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00311##
[0310] Example I-70
2-[(2S)-(isoquinolin-1-ylcarbonylamino)-3,3-dimethyl-butyryl]-2-(1S,4R)-az-
a-bicyclo[2.2.1]heptane-(3S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00312##
[0311] Example I-71
2-[(2S)-(4-Amino-3-chloro-benzoylamino)-3,3-dimethyl-butyryl]-2-(1S,4R)-az-
a-bicyclo[2.2.1]heptane-(3S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]amide
##STR00313##
[0312] Example I-72
2-[(2S)-(4-Amino-3-chloro-benzoylamino)-3-methyl-butyryl]-2-(1S,4R)-aza-bi-
cyclo[2.2.1]heptane-(3S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00314##
[0313] Example I-73
2-[(2S)-(isoquinolin-1-ylcarbonylamino)-3,3-dimethyl-butyryl]-2-(1S,4R)-az-
a-bicyclo[2.2.1]heptane-(3S)-carboxylic acid
[(2S)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide
##STR00315##
TABLE-US-00003 [0314] TABLE 3 Characterization Data for Selected
Compounds of Formula I (by Compound Number) M + 1 No. (obs)
.sup.1H-NMR I-1 490.1 (DMSO-d.sub.6) 0.94-0.95 (3H, m), 0.98-0.99
(3H, m), 1.13-1.16 (3H, m), 1.80-2.0 (4H, m), 2.10 (3H, s),
2.47-2.51 (2H, m), 2.73 (1H, m), 3.34-3.61 (2H, m), 3.73-3.77 (2H,
m), 3.79 (3H, s), 3.90 (1H, m), 4.39 (1H, m), 4.55 (1H, m), 5.55
(1H, d), 6.83 (1H, d), 6.99 (1H, d), 7.19 (1H, m), 8.27 (1H, d),
8.34 (1H, d) I-2 476.0 (CDCl.sub.3) 1.01-1.15 (6H, m), 1.26 (3H,
t), 1.90-2.29 (5H, m), 2.55-2.59 (1H, m), 2.75-2.83 (1H, m),
3.65-3.98 (3H, m), 4.04 (3H, s), 4.44-4.49 (1H, m), 4.62-4.69 (1H,
m), 4.75-4.80 (1H, m), 5.60 (1H, d), 7.09 (1H, t), 7.19 (1H, d),
7.52 (1H, t), 7.97 (1H, d) I-3 530.0 (CDCl.sub.3) 1.02-1.10 (6H,
m), 1.23-1.34 (3H, m), 1.88-2.19 (5H, m), 2.32-2.44 (2H, m),
2.81-2.89 (1H, m), 3.66-3.72 (2H, m), 3.83-3.98 (2H, m), 4.56-4.73
(2H, m), 4.84-4.90 (1H, m), 5.46 (1H, d), 7.15 (1H, d), 7.35-7.60
(4H, m), 7.99 (1H, d) I-4 476.1 (CDCl.sub.3) 1.02 (3H, d), 1.09
(3H, d), 1.29 (3H, t), 1.93-2.19 (4H, m), 2.29 (3H, s), 2.39 (2H,
dd), 2.84 (1H, dd), 3.66-3.71 (2H, m), 3.88-3.95 (2H, m), 4.63 (1H,
dd), 4.68-4.74 (1H, m), 4.85 (1H, dd), 5.32 (1H, s), 5.47 (1H, d),
6.44 (1H, d), 6.87 (1H, d), 6.98-7.00 (1H, m), 7.07-7.12 (1H, m),
7.36 (1H, d) I-5 475.0 (CDCl.sub.3) 0.95-1.10 (6H, m), 1.31 (3H,
t), 1.93-2.21 (4H, m), 2.25 (3H, s), 2.34-2.41 (2H, m), 2.80-2.88
(1H, m), 3.63-3.75 (4H, m), 3.87-3.93 (2H, m), 4.65-4.75 (2H, m),
4.82-4.88 (1H, m), 5.47 (1H, d), 6.43 (1H, d), 6.74 (1H, d), 6.81
(1H, d), 7.04 (1H, t), 7.40 (1H, d) I-6 514.4 (CDCl.sub.3)
1.03-1.05 (3H, m), 1.09-1.13 (3H, m), 1.22-1.30 (3H, m), 1.95 (1H,
m), 2.14-2.17 (2H, m), 2.44-2.51 (2H, m), 2.79 (1H, m), 3.65-3.68
(2H, m), 3.86-3.90 (2H, m), 4.12 (1H, m), 4.60-4.61 (2H, m), 4.86
(1H, m), 5.47 (1H, m), 6.4 (1H, 2 .times. d), 7.29-7.37 (3H, m),
7.54 (1H, m) I-7 461.1 (DMSO-d.sub.6) 0.95-1.01 (6H, m), 1.13-1.16
(3H, m), 1.80-2.10 (4H, m), 2.45-2.51 (5H, m), 2.74 (1H, m),
3.33-3.59 (2H, m), 3.68 (1H, m), 3.95 (1H, m), 4.38-4.44 (2H, m),
4.55 (1H, m), 5.55 (1H, m), 7.25 (1H, m), 7.62 (1H, m), 8.27 (1H,
m), 8.48 (1H, m), 8.63 (1H, m) I-8 461.1 (CDCl.sub.3) 1.02 (3H, d),
1.08 (3H, d), 1.28 (3H, t), 1.95-2.2 (4H, m), 2.4-2.5 (2H, m), 2.55
(3H, s), 2.8-2.9 (1H, m), 3.7-3.8 (2H, m), 3.85-3.95 (2H, m),
4.7-4.85 (2H, m), 4.9-4.95 (1H, m), 5.55 (1H, d), 6.6-6.65 (1H, m),
7.2-7.25 (1H, m), 7.35-7.4 (1H, m), 8.6 (1H, d), 8.7 (1H, s) I-9
465.6 (DMSO-d.sub.6) 0.93-0.95 (3H, m), 0.99-1.00 (3H, m),
1.13-1.16 (3H, m), 1.80-2.10 (4H, m), 2.40 (3H, s), 2.40-2.47 (2H,
m), 2.73 (1H, m), 3.59-3.61 (2H, m), 3.73-3.75 (2H, m), 4.37-4.43
(2H, m), 4.55 (1H, m), 5.53 (1H, d), 6.97 (1H, m), 7.59 (1H, m),
7.81 (1H, d), 8.28 (1H, d) I-10 515.0 (CDCl.sub.3) 1.01 (3H, d),
1.25 (3H, d), 1.27 (3H, t), 1.97-2.10 (2H, m), 2.14-2.26 (1H, m)
2.38 (2H, dd), 2.84 (1H, dd), 3.67-3.71 (2H, m), 3.81-3.87 (1H, m),
3.90-3.98 (1H, m), 4.58-4.61 (1H, m), 4.65-4.73 (1H, m), 4.86-4.90
(1H, dd), 5.47 (1H, d), 6.78 (1H, d), 7.23 (1H, d), 7.42 (1H, d),
8.40 (1H, d) I-11 515.0 (CDCl.sub.3) 1.01 (3H, d), 1.14 (3H, d),
1.28 (3H, t), 1.96-2.12 (2H, m), 2.17-2.23 (2H, m), 2.38 (2H, dd),
2.83 (1H, dd), 3.66-3.71 (2H, m), 3.82-3.95 (2H, m), 4.59-4.62 (1H,
m), 4.65-4.71 (1H, m), 4.91 (1H, dd), 5.47 (1H, d), 6.54 (1H, br
dd), 7.21 (1H, br dd), 8.57 (2H, s) I-12 504.4 (DMSO-d.sub.6)
0.9-1.08 (9H, s), 1.12 (3H, t), 1.75-2.00 (3H, m), 2.00-2.15 (4H,
m), 2.34-2.50 (1H, m), 2.80 (1H, m), 3.48-3.91 (7H, m), 4.40 (1H,
m), 4.46-4.70 (2H, m), 5.58 (1H, d), 7.81 (1H, d), 7.00 (1H, d),
7.19 (1H, dd), 8.07 (1H, d), 8.27 (1H, d) I-13 476.0 (CDCl.sub.3)
0.98-1.09 (6H, m), 1.90-2.05 (4H, m), 2.35-2.56 (2H, m), 2.70-2.85
(1H, m), 3.49 + 3.55 (3H, 2 .times. s), 3.55-3.67 (1H, m), 3.86
(3H, s), 4.00-4.09 (1H, m), 4.58-4.90 (3H, m), 5.34-5.37 (1H, m),
6.25 + 6.40 (1H, 2 .times. d), 6.90-7.01 (2H, m), 7.18-7.25 (1H,
m), 7.37 + 7.54 (1H, 2 .times. d) I-14 504.0 (CDCl.sub.3) 0.99-1.11
(6H, m), 1.18-1.30 (6H, m), 1.86-2.15 (4H, m), 1.28 + 1.30 (3H, 2
.times. s), 2.36-2.86 (3H, m), 3.56-3.68 (1H, m), 3.86 (3H, s),
3.87-4.05 (2H, m), 4.50-4.84 (3H, m), 5.55 + 5.59 (1H, 2 .times.
d), 6.86-7.01 (2H, m), 7.16-7.23 (1H, m), 7.37 + 7.54 (1H, 2
.times. d) I-15 504.0 (CDCl.sub.3) 0.85-1.11 (9H, m), 1.55-1.73
(2H, m), 1.89-2.20 (4H, m), 2.28 + 2.29 (3H, 2 .times. s),
2.35-2.55 (2H, m), 2.71-2.87 (1H, m), 3.48-3.76 (3H, m), 3.86 (3H,
s), 3.98-4.06 (1H, m), 4.52-4.86 (3H, m), 5.44-5.49 (1H, m), 6.24 +
6.35 (1H, 2 .times. d), 6.88-6.99 (2H, m), 7.14-7.21 (1H, m), 7.41
+ 7.55 (1H, 2 .times. d) I-16 480.5 (CDCl3) 1.0-1.15 (6H, m),
1.3-1.4 (3H, m), 1.9-2.2 (4H, m), 2.4-2.5 (2H, m), 2.8-2.9 (1H, m),
3.7-3.8 (2H, m), 3.9-4.0 (2H, m), 4.65-4.75 (2H, m), 4.88-4.92 (1H,
m), 5.5-5.52 (1H, m), 6.85-6.9 (1H, m), 7.4-7.55 (1H, m), 7.7-7.75
(1H, m) I-17 474.6 (DMSO-d.sub.6) 1.05 (9H, s), 1.15 (3H, t),
1.8-2.1 (4H, m), 2.3 (3H, s), 2.4-2.5 (1H, m), 2.7-2.8 (1H, m),
3.6-3.9 (4H, m), 4.4-4.45 (1H, m), 4.5-4.7 (2H, m), 5.55-5.6 (1H,
m), 7.2-7.4 (4H, m), 8.1 (1H, d), 8.25 (1H, d) I-18 514.5
(DMSO-d.sub.6) 0.9-1.0 (6H, m), 1.15 (3H, t), 1.8-2.1 (4H, m),
2.4-2.5 (1H, m), 2.7-2.8 (1H, m), 3.6-3.85 (3H, m), 3.9-3.95 (1H,
m), 4.4-4.6 (3H, m), 5.55-5.6 (1H, m), 7.4-7.45 (1H, m), 7.6-7.8
(3H, m), 8.22 (1H, d), 8.75 (1H, d) I-19 480.5 (CDCl.sub.3) 1.13
(9H, s), 1.90-2.20 (3H, m), 2.35-2.44 (2H, m), 2.86 (1H, dd), 3.56
(3H, s), 3.72-3.74 (1H, m), 3.90-3.99 (1H, m), 4.62-4.65 (1H, m),
4.69-4.70 (1H, m), 4.90 (1H, d), 5.36 (1H, d), 6.94 (1H, d),
7.28-7.46 (4H, m), 7.71 (1H, dd) I-20 542.5 (CDCl.sub.3) 1.09 (9H,
s), 1.27 (6H, m), 1.93-2.14 (3H, m), 2.34-2.42 (2H, m), 2.79-2.83
(1H, m), 3.71 (1H, m), 3.90-3.94 (1H, m), 4.01-4.04 (1H, m),
4.62-4.67 (2H, m), 4.88-4.91 (1H, m), 5.56 (1H, m), 6.46 (1H, m),
7.40 (1H, m), 7.54-7.62 (3H, m), 7.74 (1H, m) I-21 494.5
(CDCl.sub.3) 1.12 (9H, s), 1.29 (3H, t), 1.90-2.20 (3H, m),
2.36-2.43 (2H, m), 2.85 (1H, dd), 3.67-3.72 (2H, m), 3.90-3.96 (2H,
m), 4.62-4.65 (2H, m) 4.91 (1H, d), 5.46 (1H, d), 6.95 (1H, d),
7.34-7.46 (4H, m), 7.71 (1H, dd) I-22 528.4 (CDCl.sub.3) 1.10 (9H,
s), 1.29 (3H, t), 1.90-2.20 (3H, m), 2.35-2.42 (2H, m), 2.84 (1H,
dd), 3.68-3.72 (2H, m), 3.90-3.95 (2H, m), 4.62-4.80 (2H, m), 4.89
(1H, d), 5.47 (1H, d), 6.45 (1H, d), 7.43 (1H, d), 7.54-7.61 (3H,
m), 7.73 (1H, dd) I-23 508.5 (CDCl.sub.3) 0.95 (3H, t), 1.12 (9H,
s), 1.60-1.70 (2H, m), 1.88-2.20 (3H, m), 2.35-2.45 (2H, m),
2.77-2.85 (1H, m), 3.53-3.61 (1H, m), 3.65-3.75 (1H, m), 3.76-3.84
(1H, m), 3.88-3.96 (1H, m), 4.60-4.73 (2H, m), 4.91 (1H, d), 5.44
(1H, d), 6.96 (1H, d), 7.30-7.50 (4H, m), 7.73 (1H, d) I-24 522.5
(CDCl.sub.3) 0.86 (3H, t), 1.18 (9H, s), 1.21-1.65 (4H, m),
1.85-2.17 (3H, m), 2.36-2.59 (2H, m), 2.68-2.78 (1H, m), 3.44-3.54
(1H, m), 3.56-3.72 (2H, m), 3.98-4.10 (1H, m), 4.56-4.85 (3H, m),
5.44 (1H, d), 6.95-7.02 (1H, m), 7.32-7.74 (5H, m) I-25 578.3
(DMSO-d.sub.6) 0.99-1.21 (12H, m), 1.70-2.00 (3H, m), 2.01-2.17
(1H, m), 2.40-2.51 (1H, m), 2.70-2.80 (1H, m), 3.50-3.88 (4H, m),
4.40 (1H, m), 4.55 (1H, m), 4.65 (1H, m), 5.58 (1H, d), 7.36 (1H,
m), 7.50 (1H, m), 7.61 (1H, m), 8.21 (1H, d), 8.70 (1H, d) I-26
494.5 (CDCl.sub.3) 0.95 (3H, t), 1.05-1.15 (6H, m), 1.55-1.8 (3H,
m), 2.0-2.25 (4H, m), 2.4-2.5 (1H, m), 2.6-2.9 (2H, m), 3.55-3.8
(3H, m), 3.85-3.95 (1H, m), 4.05-4.1 (1H, m), 4.7-4.85 (2H, m),
5.5-5.55 (1H, m), 6.85-6.9 (1H, m), 7.4-7.6 (3H, m), 7.7-7.8 (1H,
m) I-27 494.5 (CDCl.sub.3) 0.95 (3H, t), 1.05-1.15 (6H, m), 1.5-1.7
(3H, m), 2.0-2.2 (4H, m), 2.4-2.6 (2H, m), 2.9-3.1 (1H, m), 3.4-3.5
(1H, m), 3.55-3.7 (2H, m), 4.0-4.1 (1H, m), 4.35-4.5 (2H, m),
4.6-4.75 (1H, m), 4.8-4.9 (0.5H, m), 5.35-5.38 (1H, m), 6.85-6.95
(1H, m), 7.4-7.55 (3H, m), 7.64-7.8 (1.5H, m) I-28 480.3 (CDCl3)
1.02-1.19 (7H, m), 1.22-1.28 (2H, m), 1.90-2.21 (3H, m), 2.32-2.53
(2H, m), 2.95 (1H, 2 .times. dd), 3.44-3.50 (1H, m), 3.59-78 (2H,
m), 3.83-3.92 (1H, m), 4.02-4.09 (1H, m), 4.29-4.41 (1H, m), 5.34
(1H, 2 .times. s), 6.88 (1H, 2 .times. brd d), 7.31-7.42 (4H, m),
7.57 (1H, 2 .times. brd d), 7.70 (1H, 2 .times. dd) I-29 508
(CDCl3) 0.83-0.97 (3H, m), 1.02-1.14 (6H, m), 1.26-1.53 (3H, m),
1.55-1.66 (1H, m), 1.91-2.20 (4H, m), 2.35-2.61 (2H, m), 2.73-2.90
(1H, m), 3.54-3.74 (3H, m), 3.84-3.90 (0.5H, m), 3.99-4.06 (0.5H,
m), 4.61-4.75 (2H, m), 4.77-4.93 (0.5H, m), 5.45-5.51 (1H, m), 6.87
(1H, brd), 7.34-7.45 (4H, m), 7.55 (0.5H, brd), 7.70-7.22 (1H, m)
I-30 508 (400 MHz, CDCl3) 0.87-0.97 (3H, m), 0.99-1.16 (6H, m),
1.27-1.40 (2H, m), 1.48-1.59 (1H, m), 1.91-2.19 (4H, m), 2.30-2.52
(2H, m), 2.90-3.07 (1H, m), 3.39-3.45 (0.5H, m), 3.54-3.71 (2H, m),
3.78-3.82 (0.5H, m), 3.86-3.92 (0.5H, m), 4.04-4.09 (0.5H, m),
4.31-4.35 (1H, m), 4.39-4.43 (1H, m), 4.56-4.59 (0.5H, m),
4.66-4.68 (1H, m), 4.80-4.86 (0.5H, m), 5.32-5.41 (1H, m),
6.87-6.91 (1H, m), 7.31-7.45 (4H, m), 7.55-7.76 (2H, m) I-31 494.4
(CDCl3) 1.04-1.19 (8H, m), 1.25-1.28 (3H, m), 1.92-2.18 (4H, m),
2.32-2.43 (1H, m), 2.62-2.87 (2H, m), 3.59-3.71 (1H, m), 3.85-3.95
(1H, m), 4.00-4.05 (1H, m), 4.60-4.67 (3H, m), 5.60 (1H, 2 .times.
d), 6.88 (1H, brd d), 7.36-7.50 (4H, m), 7.52-7.56 (1H, m), 7.76
(1H, 2 .times. dd) I-32 494.3 (CDCl3) 0.87-1.24 (10H, m), 1.88-2.07
(3H, m), 2.13-2.21 (1H, m), 2.32-2.54 (2H, m), 2.94 (1H, 2 .times.
dd), 3.57-3.68 (1H, m), 3.83-3.87 (1H, m), 4.02-4.09 (1H, m),
4.27-4.30 (1H, m), 4.41 (1H, dd), 4.51-4.69 (1H, m), 5.43 (1H, 2
.times. s), 6.89 (1h, 2 .times. brd d), 7.30-7.45 (4H, m), 7.52
(1H, 2 .times. brd d), 7.70 (1H, 2 .times. dd)
I-33 550.5 (DMSO) 0.70 (2H, m, CH2), 0.89 (2H, m, CH2), 0.95-1.20
(12H, m, CH3, tbutyl), 1.71-2.13 (4H, m, CH2), 2.45 (1H, m, asp
CH2), 2.75 (1H, m, asp CH2), 3.35-3.89 (4H, m, CH2, CH), 3.99 (1H,
m, CH), 4.37 (1H, m, CH), 4.51 (1H, m, CH), 4.65 (1H, m, CH), 5.58
(1H, d, CHO), 6.90 (1H, m, aryl H), 7.35 (1H, m, aryl H), 7.45 (1H,
m, aryl H), 8.25 (1H, d, NH), 8.35 (1H, d, NH) I-34 508.5 DMSO)
0.99-1.21 (12H, m, CH3, tBu), 1.75-2.14 (4H, m, CH2), 2.38 (3H, s,
CH3), 2.40-2.51 (1H, m, asp CH2), 2.70-2.82 (1H, m, asp CH2),
3.37-3.90 (4H, m, CH2, CH), 4.39 (1H, m, CH), 4.55 (1H, m, CH),
4.67 (1H, m, CH), 5.58 (1H, d, CH), 7.15 (1H, m, aryl H), 7.28 (1H,
m, aryl H), 7.38 (1H, m, aryl H), 8.25 (1H, m, NH), 8.38 (1H, m,
NH) I-35 510 CDCl3 1.00 (3H, d), 1.10 (3H, d), 1.27 (3H, t),
1.90-2.19 (4H, m), 2.34-2.45 (2H, m), 2.79-2.87 (1H, m), 3.65-3.71
(2H, m), 3.84-4.93 (2H, m), 3.92 (3H, s), 4.56-4.70 (2H, m),
4.82-4.88 (1H, m), 4.45 (1H, d), 6.69 (1H, d), 6.99 (1H, d), 7.16
(1H, d), 7.27 (1H, t), 7.37 (1H, d) I-36 522.5 (DMSO) 0.95-1.25
(15H, m, tBu, CH3), 1.78-2.13 (4H, m, CH2), 2.43 (1H, m, CH2),
2.65-2.80 (3H, m, CH2), 3.50-3.88 (4H, m, CH2, CH), 4.42 (1H, m,
CH), 4.58 (1H, m, CH), 4.70 (1H, d, CH), 5.58 (1H, d, CH), 7.15
(1H, m, aryl H), 7.27 (1H, m, aryl H), 7.38 (1H, m, aryl H), 8.27
(1H, d, NH), 8.39 (1H, d, NH) I-37 510.5 CDCl3 1.05-1.12 (6H, m),
1.25-1.3 (3H, m), 1.9-2.2 (2H, m), 2.4-2.5 (2H, m), 2.8-2.9 (1H,
m), 3.65-3.75 (2H, m), 3.85 (3H, s), 3.9-4.0 (1H, m), 4.65-4.75
(2H, m), 4.85-4.9 (1H, m), 6.9-6.93 (1H, m), 6.98 (1H, s), 7.05-7.1
(1H, m), 7.4-7.45 (1H, m), 7.75-7.8 (1H, d) I-38 564 CDCl3
0.38-0.42 (2H, m), 0.63-0.71 (2H, m), 1.11 (9H, s), 1.23-1.35 (4H,
m), 1.88-2.20 (3H, m), 2.34-2.45 (2H, m), 2.76-2.87 (1H, m),
3.66-3.75 (2H, m), 3.87-3.96 (4H, m), 4.62-4.73 (2H, m), 4.89 (1H,
d), 5.47 (1H, d), 6.80 (1H, d), 7.00 (1H, d), 7.19-7.29 (2H, m),
7.48 (1H, d) I-39 510 (DMSO) 1.11 (9H, s), 1.28 (3H, t), 1.83-2.22
(3H, m), 2.36-2.43 (2H, m), 2.82-2.87 (1H, m), 3.66-3.76 (2H, m),
3.86-3.97 (2H, m), 4.62-4.71 (2H, m), 4.88 (1H, d), 5.45 (1H, d),
6.31 (1H, s), 6.73 (1H, d), 7.05-7.20 (3H, m), 7.38 (1H, d) I-40
537.4 (CDCl3) 1.06 (6H, dd), 1.28-1.31 (4H, m), 1.91-2.20 (4H, m),
2.23 (3H, s), 2.39 (1H, dd), 2.84 (1H, dd), 3.65-3.72 (2H, m),
3.86-3.94 (2H, m), 4.61-4.73 (2H, m), 4.87 (1H, dd), 5.46 (1H, dd),
7.00-7.04 (1H, m), 7.22 (1H, brd s), 7.38-7.45 (2H, m), 7.73 (1H,
d), 7.80 (1H, brd s) I-41 551.5 (DMSO) 0.95-1.20 (12H, m, tBu,
CH3), 2.75-2.15 (7H, m, CH2, COCH3), 2.42 (1H, m, CH2), 2.77 (1H,
m, CH2), 3.50-3.88 (4H, m, CH2, CH), 4.37 (1H, m, CH), 4.55 (1H, m,
CH), 4.67 (1H, d, CH), 5.58 (1H, d, CH), 7.09 (1H, m, aryl H), 7.32
(1H, m, aryl H), 7.71 (1H, m, aryl H), 8.26 (1H, m, NH), 8.49 (1H,
m, NH), 9.58 (1H, m, NH) I-42 531.6 (DMSO) 0.95-1.20 (12H, m, tBu,
CH3), 1.75-2.17 (10H, m, CH3, COCH3, CH2), 2.45 (1H, m, CH2), 2.77
(1H, m, CH2), 3.48-3.91 (4H, m, CH2, CH), 4.31-4.70 (3H, m, CH),
5.55 (1H, d, CH), 7.04 (1H, m, aryl H), 7.18 (1H, m, aryl H), 7.41
(1H, m, aryl H), 8.20 (1H, d, NH), 8.27 (1H, d, NH), 9.39 (1H, brs,
NH) I-43 551.4 DMSO) 1.04 (9H, s), 1.12-1.17 (3H, m), 1.78-1.95
(4H, m), 2.06 (3H, s), 2.45 (1H, dd), 2.72 (1H, dd), 3.52-3.81 (4H,
m), 4.36-4.39 (1H, m), 4.47-4.54 (1H, m), 4.64 (1H, d), 5.54 (1H,
dd), 7.33-7.35 (1H, m), 7.43-7.46 (1H, m), 7.81 (1H, brd s),
8.21-8.25 (2H, m), 10.23 (1H, brd s) I-44 535.4 (DMSO) 1.02 (9H,
s), 1.14 (3H, t), 1.78-1.98 (4H, m), 2.08 (3H, s), 2.48 (1H, dd),
2.79 (1H, dd), 3.51-3.82 (4H, m), 4.36-4.39 (1H, m), 4.49-4.58 (1H,
m), 4.71 (1H, d), 5.54 (1H, d), 7.31-7.34 (1H, m), 7.65-7.72 (3H,
m), 8.49 (1H, d), 10.38 (1H, s) I-45 521.4 (DMSO) 0.95 (6H, dd),
1.12-1.16 (4H, m), 1.72-1.97 (4H, m), 2.07 (3H, s), 2.48 (1H, dd),
2.73 (1H, dd), 3.51-3.62 (2H, m), 3.71-3.83 (2H, m), 4.35-4.38 (1H,
m), 4.48-4.59 (2H, m), 5.53 (1H, d), 7.29-7.31 (1H, m), 7.59-7.67
(2H, m), 8.01-8.05 (1H, m), 8.28 (1H, d), 10.35 (1H, s) I-46 538.5
(CDCl3 1.1-1.12 (6H, m), 1.3 (3H, m), 1.4 (6H, d), 2.0-2.2 (2H, m),
2.4-2.5 (2H, m), 2.8-2.9 (1H, m), 3.7-3.75 (2H, m), 3.9-4.0 (1H,
m), 4.6-4.75 (3H, m), 4.85-4.95 (1H, m), 6.85-6.9 (1H, m), 6.95
(1H, s), 7.05-7.1 (1H, m), 7.4-7.45 (1H, m), 7.8 (1H, d) I-47 510.5
(CDCl3) 1.15 (9H, m), 1.25 (3H, t), 2.0-2.2 (4H, m), 2.4-2.5 (2H,
m), 2.8-2.9 (1H, m), 3.7-3.85 (2H, m), 3.9-4.0 (1H, m), 4.05-4.1
(1H, m), 4.7-4.8 (1H, m), 4.85 (1H, d), 5.5 (1H, m), 6.5 (1H, d),
6.8 (1H, s), 7.2 (1H, d), 7.4 (1H, d), 7.55 (1H, d) I-48 538.5
(CDCl3) 1.12 (9H, s), 1.29 (3H, t), 1.90-2.20 (3H, m), 2.36-2.43
(2H, m), 2.85 (1H, m), 3.42 (3H, s), 3.68-3.74 (2H, m), 3.91-3.95
(2H, m), 4.48 (2H, s), 4.62-4.75 (2H, m), 4.90 (1H, m), 5.47 (1H,
m), 7.00 (1H, m), 7.31 (1H, m), 7.43-7.54 (2H, m), 7.72 (1H, m)
I-49 579.5 (CDCl3) 1.12 (9H, s), 1.28-1.31 (9H, m), 1.90-2.20 (3H,
m), 2.36-2.43 (2H, m), 2.54 (1H, m), 2.85 (1H, m), 3.68-3.72 (2H,
m), 3.91-3.95 (2H, m), 4.62-4.69 (2H, m), 4.88 (1H, d), 5.47 (1H,
m), 7.14 (1H, d), 7.27 (1H, m), 7.41 (1H, m), 7.50 (1H, d), 7.78
(1H, d), 7.87 (1H, m) I-50 577.3 (DMSO) 1.12-1.16 (7H, m),
1.58-1.81 (5H, m), 1.83-1.92 (5H, m), 2.04-2.08 (4H, m), 2.50 (1H,
dd), 2.75 (1H, dd), 3.57-3.66 (2H, m), 3.72-3.78 (1H, m), 3.82-3.91
(1H, m), 4.33-4.36 (1H, m), 4.46 (1H, t), 4.52-4.61 (1H, m), 5.54
(1H, d), 7.32 (1H, d), 7.43 (1H, dd), 7.81 (1H, d), 8.25 (1H, d),
8.47 (1H, d), 10.22 (1H, s) I-51 567.4 (DMSO) 0.98-1.25 (12H, m,
tBu, CH3), 1.78-2.14 (4H, m, CH2), 2.44 (1H, m, CH2), 2.78 (1H, m,
CH2), 3.50-3.88 (7H, m, CH3, CH2, CH), 4.38 (1H, m, CH), 4.55 (1H,
m, CH), 4.67 (1H, d, CH), 5.58 (1H, d, CH), 7.30-7.42 (2H, m, aryl
H), 7.60 (1H, brs, NH), 8.21 (2H, m, aryl H, NH), 9.99 (1H, brs,
NH). I-52 586.4 (DMSO) 0.95-1.24 (12H, m, tBu, CH3), 1.70-2.13 (4H,
m, CH2), 2.44 (1H, m, CH2), 2.75 (1H, m, CH2), 3.45-3.90 (4H, m,
CH2, CH), 4.37 (1H, m, CH), 4.55 (1H, m, CH), 4.70 (1H, d, CH),
5.57 (1H, d, CH), 6.91 (2H, d, aryl H), 7.06-7.19 (3H, m, aryl H),
7.30-7.45 (3H, m, aryl H), 8.20 (1H, d, NH), 8.55 (1H, d, NH) I-53
578.5 (DMSO) 0.9-1.0 (6H, m), 1.18 (3H, t), 1.8-2.15 (4H, m),
2.4-2.5 (1H, m), 2.7-2.8 (1H, m), 3.6-3.85 (4H, m), 4.4-4.6 (3H,
m), 5.55 (1H, d), 7.05 (1H, d), 7.3-7.35 (2H, m), 7.98 (1H, s), 8.3
(1H, d), 8.45 (1H, d), 10.7 (1H, s) I-54 495.0 (DMSO) 0.94-0.98
(6H, m), 1.13-1.18 (3H, m), 1.80-2.10 (5H, m), 2.50 (1H, m), 2.73
(1H, m), 3.58-3.61 (2H, m), 3.74 (1H, m), 3.9 (1H, m), 4.38-4.41
(2H, m), 4.60 (1H, m), 5.46 (2H, s), 5.54 (1H, m), 6.48 (1H, m),
6.80 (1H, m), 7.04 (1H, m), 8.27 (1H, d), 8.40 (1H, d) I-55 535.0
(CDCl3) 1.25 (3H, t), 1.99-2.01 (3H, s), 2.30-2.39 (1H, m), 2.68
(1H, dd), 2.79 (1H, dd), 3.21-3.27 (1H, m), 3.39 (1H, dd),
3.47-3.51 (2H, m), 3.65-3.75 (1H, m), 3.88-3.94 (1H, m), 4.64-4.68
(1H, m), 4.70-4.78 (1H, m), 5.56 (1H, d), 7.31-7.35 (5H, m),
7.63-7.65 (1H, m), 8.00 (1H, d), 8.76 (1H, d) I-56 545.0 (CDCl3)
1.25 (3H, t), 2.01-2.03 (3H, m), 2.25 (3H, s), 2.30-2.37 (1H, m),
2.65 (1H, dd), 2.80 (1H, dd), 3.27-3.41 (2H, m), 3.47 (1H, dd),
3.65-3.79 (2H, m), 3.85 (3H, s), 3.86-3.90 (1H, m), 4.64-4.67 (1H,
m), 4.71-4.80 (1H, m), 5.18-5.22 (1H, m), 5.54 (1H, d), 6.83 (1H,
d), 6.90-6.97 (2H, m), 7.19 (1H, t), 7.24-7.28 (1H, m), 7.90 (1H,
d), 8.77 (1H, d) I-57 524.0 (CDCl3) 1.12 (9H, s), 1.31 (3H, t),
1.93-2.20 (3H, m), 2.35-2.46 (2H, m), 2.79-2.86 (1H, m), 3.65-3.74
(2H, m), 3.87-3.96 (2H, m), 3.95 (3H, s), 4.65-4.74 (2H, m), 4.89
(1H, d), 5.47 (1H, d), 6.76 (1H, d), 7.03 (1H, d), 7.30 (1H, t),
7.48 (1H, d) I-58 530.4 (CDCl3) 1.10 (9H, s), 1.28 (3H, t),
1.43-1.56 (1H, m), 1.79-1.86 (3H, m), 1.99 (1H, brd), 2.29 (3H, s),
2.30-2.37 (1H, m), 2.83 (1H, dd), 3.02 (1H, brs), 3.66-3.74 (1H,
m), 3.87 (3H, s), 3.88-3.94 (1H, m), 4.16 (1H, brs), 4.54 (1H,
brs), 4.66-4.74 (1H, m), 4.97 (1H, d), 5.46 (1H, d), 6.44 (1H,
brd), 6.93 (1H, d), 7.00 (1H, d), 7.22 (1H, t), 7.78 (1H, brd) I-59
520.5 (CDCl3) 1.13 (9H, s), 1.29 (3H, t), 1.76-1.90 (3H, m), 2.00
(1H, brd), 2.35 (1H, dd), 2.83 (1H, dd), 3.66-3.74 (1H, m),
3.87-3.94 (1H, m), 4.15 (1H, s), 4.54 (1H, brs), 4.62-4.78 (1H, m),
4.99 (1H, d), 5.46 (1H, d), 6.92 (1H, brd), 7.33-7.46 (3H, m), 7.69
(1H, brdd), 7.77 (1H, brd) I-60 577.5 (CDCl3) 1.12 (9H, s),
1.26-1.31 (3H, m), 1.43-1.45 (1H, m), 1.83 (3H, brs), 1.99 (1H,
brd), 2.06 (1H, m), 2.23 (3H, s), 2.34 (1H, brdd), 2.83 (1H, brdd),
3.01 (1H, brs), 3.66-3.74 (1H, m), 3.87-3.95 (1H, m), 4.12-4.19
(1H, m), 4.53 (1H, brs), 4.65-4.76 (1H, m), 4.98 (1H, d), 5.45-5.47
(1H, m), 7.08 (1H, brd), 7.30 (1H, m), 7.37 (1H, brd), 7.73-7.75
(1H, m), 7.80-7.82 (2H, m) I-61 591.5 (CDCl3) 1.14 (9H, s),
1.22-1.30 (6H, m), 1.54-1.57 (1H, m), 1.77-1.85 (3H, m), 1.97 (1H,
d), 2.30-2.45 (3H, m), 2.75-2.84 (1H, m), 3.00 (1H, s), 3.63-3.72
(1H, m), 3.84-3.93 (1H, m), 4.10-4.16 (1H, m), 4.51 (1H, s),
4.64-4.71 (1H, m), 4.96 (1H, d), 5.45 (1H, d), 7.05 (1H, d), 7.26
(1H, s), 7.36 (1H, d), 7.73 (1H, d), 7.80 (1H, d), 7.82 (1H, s)
I-62 605.6 (CDCl3) 1.15 (9H, s), 1.3 (3H, t), 1.35 (6H, d),
1.4-1.55 (3H, m), 1.8-1.95 (3H, m), 2.0-2.1 (1H, m), 2.3-2.4 (1H,
m), 2.65-2.75 (1H, m), 2.8-2.9 (1H, m), 3.05 (1H, s), 3.7-3.8 (1H,
m), 3.9-4.0 (1H, m), 4.2 (1H, s), 4.55 (1H, s), 4.7-4.8 (1H, m),
5.0 (1H, d), 5.5 (1H, d), 6.6 (1H, d), 7.3-7.45 (2H, m), 7.75 (1H,
d), 7.85 (1H, s), 8.55 (1H, d) I-63 534.4 (CDCl3) 1.13 (9H, s),
1.31 (3H, t), 1.42-1.48 (1H, m), 1.56 (1H, brs), 1.77-1.83 (3H, m),
1.99 (1H, brd),
2.35 (1H, dd), 2.83 (1H, dd), 3.01 (1H, brs), 3.67-3.76 (1H, m),
3.88-3.99 (4H, m), 4.14 (1H, brs), 4.52 (1H, brs), 4.65-4.73 (1H,
m), 5.00 (1H, dd), 5.47 (1H, d), 7.10-7.21 (2H, m), 7.34-7.39 (1H,
m), 7.56-7.61 (1H, m), 7.89 (1H, d) I-64 520.5 (CDCl3) 1.03 (3H,
d), 1.10 (3H, d), 1.32 (3H, t), 1.50 (1H, m), 1.59 (1H, m),
1.812-1.84 (3H, m), 2.0 (1H, m), 2.15 (1H, m), 2.36 (1H, m), 2.83
(1H, m), 3.02 (1H, br s), 3.69 (1H, m), 3.90-3.95 (4H, m), 4.13
(1H, br s), 4.40 (1H, br s), 4.67 (1H, m), 4.97 (1H, m), 5.47 (1H,
d), 7.12-7.21 (2H, m), 7.28 (1H, m), 7.59 (1H, m), 7.80 (1H, m)
I-65 530.9 (DMSO) 0.91-2.40 (23H, m), 2.95-3.40 (2H, m), 3.51-3.81
(5H, m), 4.00-4.71 (3H, m), 5.29 (1H, m), 6.80 (1H, d), 7.00 (1H,
d), 7.19 (1H, t), 7.94 (1H, d), 8.48 (1H, d) I-66 522.8 (DMSO)
0.95-1.20 (12H, m), 1.24-1.40 (2H, m), 1.41-2.40 (6H, m), 3.05 (1H,
m), 3.50-3.80 (3H, m), 4.15 (1H, m), 4.60 (1H, m), 4.70 (1H, d),
5.30 (1H, s), 7.28-7.50 (4H, m), 8.35 (1H, d), 8.48 (1H, d) I-67
577.5 .sup.(CDCl3) .delta. 1.10 (9H, s), 1.26-1.33 (3H, m),
1.43-1.45 (1H, m), 1.74-1.83 (2H, m), 2.01 (1H, brd), 2.06 (1H, m),
2.30 (3H, s), 2.37 (1H, brdd), 2.85 (1H, brdd), 2.99 (1H, brs),
3.69-3.76 (1H, m), 3.89-3.97 (1H, m), 4.11-4.31 (2H, m), 4.53 (1H,
brs), 4.65-4.76 (1H, m), 4.95 (1H, d), 5.45-5.47 (1H, m), 6.75 (1H,
brd), 7.67-7.69 (2H, m), 7.78 (1H, brs), 7.92 (1H, m), 8.55 (1H,
brd) I-68 577.5 (CDCl3) 1.10 (9H, s), 1.26-1.33 (3H, m), 1.43-1.45
(1H, m), 1.74-1.83 (2H, m), 2.01 (1H, brd), 2.06 (1H, m), 2.30 (3H,
s), 2.37 (1H, brdd), 2.85 (1H, brdd), 2.99 (1H, brs), 3.69-3.76
(1H, m), 3.89-3.97 (1H, m), 4.11-4.31 (2H, m), 4.53 (1H, brs),
4.65-4.76 (1H, m), 4.95 (1H, d), 5.45-5.47 (1H, m), 6.75 (1H, brd),
7.67-7.69 (2H, m), 7.78 (1H, brs), 7.92 (1H, m), 8.55 (1H, brd)
I-69 591.5 (CDCl3) 1.10 (9H, s), 1.26-1.33 (6H, m), 1.42-1.16 (1H,
m), 1.55-1.83 (4H, m), 2.01 (1H, brd), 2.36 (1H, dd), 2.53 (2H, q),
2.83 (1H, dd), 2.99 (1H, brs), 3.69-3.76 (1H, m), 3.89-3.96 (1H,
m), 4.11 (1H, s), 4.53 (1H, brs), 4.66-4.77 (1H, m), 4.95 (1H, d),
5.48 (1H, d), 6.76 (1H, d), 7.67-7.74 (2H, m), 8.80 (1H, s), 7.90
(1H, d), 8.58 (1H, d) I-70 537.4 (CDCl3) 1.12 (9H, s), 1.23-1.30
(3H, m), 1.36-1.41 (1H, m), 1.73-1.84 (3H, m), 1.98-2.03 (1H, m),
2.33-2.41 (1H, m), 2.75-2.83 (1H, m), 2.96 (1H, brs), 3.65-3.73
(1H, m), 3.84-3.93 (1H, m), 4.11 (1H, brs), 4.56 (1H, s), 4.63-4.71
(1H, m), 4.96-4.99 (1H, m), 5.43-5.46 (1H, m), 7.64-7.72 (2H, m),
7.79-7.87 (3H, m), 8.48-8.52 (1H, m), 8.90 (1H, brd), 9.51 (1H, d)
I-71 535.6 (CDCl3) 1.09 (9H, s), 1.32 (3H, t), 1.41-1.71 (5H, m),
1.76-1.87 (3H, m), 2.00 (1H, brd), 2.37 (1H, dd), 2.83 (1H, dd),
2.98 (1H, brs), 3.68-3.77 (1H, m), 3.89-3.97 (1H, m), 4.11 (1H, s),
4.54 (1H, brs), 4.67-4.74 (1H, m), 4.95 (1H, d), 5.48 (1H, d), 6.64
(1H, brd), 6.78 (1H, d), 7.54 (1H, dd), 7.71 (1H, brd), 7.78 (1H,
d) I-72 521.5 (CDCl3) 1.05 (3H, d), 1.15 (3H, d), 1.35 (3H, t),
1.5-1.6 (1H, m), 1.6-1.7 (1H, m), 1.8-1.9 (2H, s), 2.0-2.05 (1H,
m), 2.15-2.25 (1H, m), 2.35-2.45 (1H, m), 2.8-2.9 (1H, m), 2.95
(1H, s), 3.7-3.8 (1H, m), 3.9-4.0 (1H, m), 4.1 (1H, s), 4.45 (3H,
s), 4.7-4.8 (1H, m), 4.9-4.95 (1H, m), 5.55 (1H, d), 6.7 (1H, d),
6.85 (1H, d), 7.65 (1H, d), 7.75 (1H, d), 7.82 (1H, s) I-73 537.4
(CDCl3) 1.14 (9H, s), 1.25 (3H, t), 1.40-1.46 (1H, m), 1.77-1.89
(3H, m), 1.98-2.02 (1H, m), 2.34 (1H, dd), 2.99-3.05 (1H, m),
3.62-3.69 (1H, m), 3.83-3.91 (1H, m), 4.12 (1H, s), 4.29-4.34 (1H,
m), 4.59 (1H, s), 4.99 (1H, d), 5.38 (1H, s), 7.67-7.76 (2H, m),
7.86 (2H, dd), 8.13 (1H, d), 8.56 (1H, d), 8.96 (1H, d), 9.56 (1H,
d)
Example II-1
(S,S,S)-(3S)-({1-[(2S)-(3-Methoxy-2-methyl-benzoylamino)-3-methyl-butyryl]-
-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00316##
[0315] Method I
[0316]
(S,S,S,R)-1-[(2S)-(3-Methoxy-2-methyl-benzoylamino)-3-methyl-butyry-
l]-pyrrolidine-(2S)-carboxylic acid
[(2R)-ethoxy-5-oxo-tetrahydro-furan-(3S)-yl]-amide (97.6 mg, 0.20
mmol) was dissolved in a mixture of 2M HCl (2 ml) and MeCN (2 ml).
The reaction mixture was stirred at room temperature for 2.5 hours.
The resulting crude mixture was diluted with EtOAc and washed with
water. The aqueous layer was extracted twice with EtOAc. The
combined organic extracts were washed with brine, dried over
magnesium sulfate, filtered and concentrated in vacuo. The residue
was co-evaporated with DCM/Petrol to afford the title compound as a
white solid (81.3 mg, 88% yield).
[0317] Compounds of formula II-2 to II-61 have been prepared by
methods substantially similar to those described in Example
II-1.
Example II-2
(S,S,S)-(3S)-({1-[(2S)-(2-Chloro-benzoylamino)-3-methyl-butyryl]-pyrrolidi-
ne-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00317##
[0318] Example II-3
(S,S,S)-(3S)-({1-[3-Methyl-(2S)-(2-methyl-benzoylamino)-butyryl]-pyrrolidi-
ne-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00318##
[0319] Example II-4
(S,S,S)-(3S)-({1-[(2S)-(2-methoxy-benzoylamino)-3-methyl-butyryl]-pyrrolid-
ine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00319##
[0320] Example II-5
(S,S,S)-(3S)-({1-[3-Methyl-(2S)-(2-trifluoromethoxy-benzoylamino)-butyryl]-
-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00320##
[0321] Example II-6
(S,S,S)-(3S)-({1-[(2S)-(3-Hydroxy-2-methyl-benzoylamino)-3-methyl-butyryl]-
-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00321##
[0322] Example II-7
(S,S,S)-(3S)-({1-[(2S)-(3-Amino-2-methyl-benzoylamino)-3-methyl-butyryl]-p-
yrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00322##
[0323] Example II-8
(S,S,S)-(3S)-({1-[(2S)-(2,3-Dichloro-benzoylamino)-3-methyl-butyryl]-pyrro-
lidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00323##
[0324] Example II-9
(S,S,S)-(3S)-({1-[(2S)-(2-Chloro-3-trifluoromethyl-benzoylamino)-3-methyl--
butyryl}-pyrrolidine-(2S)-carbonyl]-amino)-4-oxo-butyric acid
##STR00324##
[0325] Example II-10
(S,S,S)-(3S)-({1-[(2S)-(3-Chloro-2-methyl-benzoylamino)-3-methyl-butyryl]--
pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00325##
[0326] Example II-11
(S,S,S)-(3S)-({1-[(2S)-(2,4-Dichloro-benzoylamino)-3-methyl-butyryl]-pyrro-
lidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00326##
[0327] Example II-12
(S,S,S)-(3S)-({1-[(2S)-(2,5-Dichloro-benzoylamino)-3-methyl-butyryl]-pyrro-
lidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00327##
[0328] Example II-13
(S,S,S)-(3S)-({1-[(2S)-(2,6-Dichloro-benzoylamino)-3-methyl-butyryl]-pyrro-
lidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00328##
[0329] Example II-14
(S,S,S)-(3S)-({1-[(2S)-(2,6-Methyl-benzoylamino)-3-methyl-butyryl]-pyrroli-
dine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00329##
[0330] Example II-15
(S,S,S)-(3S)-[(1-{3-Methyl-(2S)-[(2-methyl-pyridine-3-carbonyl)-amino]-but-
yryl}-pyrrolidine-(2S)-carbonyl)-amino]-4-oxo-butyric acid
##STR00330##
[0331] Example II-16
(S,S,S)-(3S)-[(1-{3-Methyl-(2S)-[(4-methyl-pyridine-3-carbonyl)-amino]-but-
yryl}-pyrrolidine-(2S)-carbonyl)-amino]-4-oxo-butyric acid
##STR00331##
[0332] Example II-17
(S,S,S)-(3S)-[(1-{3-Methyl-(2S)-[(3-methyl-thiophene-2-carbonyl)-amino]-bu-
tyryl}-pyrrolidine-(2S)-carbonyl)-amino]-4-oxo-butyric acid
##STR00332##
[0333] Example II-18
(S,S,S)-(3S)-[(1-{(2S)-[(2,3-Dichloro-pyridine-4-carbonyl)-amino]-3-methyl-
-butyryl}-pyrrolidine-(2S)-carbonyl)-amino]-4-oxo-butyric acid
##STR00333##
[0334] Example II-19
(S,S,S)-(3S)-[(1-{(2S)-[(3,5-Dichloro-pyridine-4-carbonyl)-amino]-3-methyl-
-butyryl}-pyrrolidine-(2S)-carbonyl)-amino]-4-oxo-butyric acid
##STR00334##
[0335] Example II-20
(S,S,S)-(3S)-({1-[(2S)-(3-Methoxy-2-methyl-benzoylamino)-3,3-dimethyl-buty-
ryl]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00335##
[0336] Example II-21
(S,S,S)-4-Oxo-(3S)-({1-[4,4,4-trifluoro-(2S)-(2-methyl-3-methoxy-benzoylam-
ino)-butyryl]-pyrrolidine-(2S)-carbonyl}-amino)-butyric acid
##STR00336##
[0337] Example II-22
(S,S,S)-(3S)-({1-[(2S)-(5-Methoxy-2-methyl-benzoylamino)-3-methyl-butyryl]-
-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00337##
[0338] Example II-23
(S,S,S)-(3S)-({1-[(2S)-(3-Methoxy-2-methyl-benzoylamino)-3-thiazol-4-yl-pr-
opionyl]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00338##
[0339] Example II-24
(S,S,S)-(3S)-({1-[(2S)-(2-Chloro-benzoylamino)-4,4,4-trifluoro-butyryl]-py-
rrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00339##
[0340] Example II-25
(S,S,S)-(3S)-({1-[(2S)-(2-Chloro-benzoylamino)-3-thiazol-4-yl-propionyl]-p-
yrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00340##
[0341] Example II-26
(S,S,S)-(3S)-({1-[3,3-Dimethyl-(2S)-(2-methyl-benzoylamino)-butyryl]-pyrro-
lidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00341##
[0342] Example II-27
(S,S,S)-(3S)-({1-[3-Methyl-(2S)-(2-trifluoromethyl-benzoylamino)-butyryl]--
pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00342##
[0343] Example II-28
(S,S,S)-(3S)-({1-[(2S)-(2-Chloro-benzoylamino)-3,3-dimethyl-butyryl]-pyrro-
lidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00343##
[0344] Example II-29
(S,S,S)-(3S)-({1-[3,3-Dimethyl-(2S)-(2-trifluoromethyl-benzoylamino)-butyr-
yl]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00344##
[0345] Example II-30
(S,S,S)-(3S)-({1-[(2S)-(2-Chloro-3-methoxy-benzoylamino)-3,3-dimethyl-buty-
ryl]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00345##
[0346] Example II-31
(S,S,S)-(3S)-({1-[(2S)-(2-Fluoro-3-methoxy-benzoylamino)-3,3-dimethyl-buty-
ryl]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00346##
[0347] Example II-32
(S,S,S)-(3S)-({1-[(2S)-(2-Chloro-3-trifluoromethoxy-benzoylamino)-3,3-dime-
thyl-butyryl]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric
acid
##STR00347##
[0348] Example II-33
(S,S,S)-(3S)-({1-[(2S)-(2-Chloro-3-cyclopropyloxy-benzoylamino)-3,3-dimeth-
yl-butyryl]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric
acid
##STR00348##
[0349] Example II-34
(S,S,S)-(3S)-({1-[(2S)-(2-Chloro-3-methyl-benzoylamino)-3,3-dimethyl-butyr-
yl]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00349##
[0350] Example II-35
(S,S,S)-(3S)-({1-[(2S)-(2-chloro-3-methoxy-benzoylamino)-3-methyl-butyryl]-
-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00350##
[0351] Example II-36
(S,S,S)-(3S)-({1-[(2S)-(2-Chloro-3-ethyl-benzoylamino)-3,3-dimethyl-butyry-
l]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00351##
[0352] Example II-37
(S,S,S)-(3S)-({1-[(2S)-(2-chloro-4-methoxy-benzoylamino)-3-methyl-butyryl]-
-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00352##
[0353] Example II-38
(S,S,S)-(3S)-({1-[(2S)-(2-Chloro-3-cyclopropylmethoxy-benzoylamino)-3,3-di-
methyl-butyryl]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric
acid
##STR00353##
[0354] Example II-39
(S,S,S)-(3S)-({1-[(2S)-(2-Chloro-3-hydroxy-benzoylamino)-3,3-dimethyl-buty-
ryl]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00354##
[0355] Example II-40
(S,S,S)-(3S)-({1-[(2S)-(2-chloro-4-acetamido-benzoylamino)-3-methyl-butyry-
l]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00355##
[0356] Example II-41
(S,S,S)-(3S)-({1-[(2S)-(2-Chloro-3-acetamido-benzoylamino)-3,3-dimethyl-bu-
tyryl]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00356##
[0357] Example II-42
(S,S,S)-(3S)-({1-[(2S)-(2-methyl-3-acetamido-benzoylamino)-3,3-dimethyl-bu-
tyryl]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00357##
[0358] Example II-43
(S,S,S)-(3S)-({1-[(2S)-(2-chloro-4-acetamido-benzoylamino)-3,3-dimethyl-bu-
tyryl]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00358##
[0359] Example II-44
(S,S,S)-(3S)-({1-[(2S)-(2-fluoro-4-acetamido-benzoylamino)-3,3-dimethyl-bu-
tyryl]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00359##
[0360] Example II-45
(S,S,S)-(3S)-({1-[(2S)-(2-fluoro-4-acetamido-benzoylamino)-3-methyl-butyry-
l]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00360##
[0361] Example II-46
(S,S,S)-(3S)-({1-[(2S)-(2-chloro-4-isopropyloxy-benzoylamino)-3-methyl-but-
yryl]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00361##
[0362] Example II-47
(S,S,S)-(3S)-({1-[(2S)-(2-chloro-4-hydroxy-benzoylamino)-3,3-dimethyl-buty-
ryl]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00362##
[0363] Example II-48
(S,S,S)-(3S)-({1-[(2S)-(2-chloro-4-methoxymethyl-benzoylamino)-3,3-dimethy-
l-butyryl]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00363##
[0364] Example II-49
(S,S,S)-(3S)-({1-[(2S)-(2-chloro-4-isobutyrylamido-benzoylamino)-3,3-dimet-
hyl-butyryl]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric
acid
##STR00364##
[0365] Example II-50
(S,S,S)-(3S)-({1-[(2S)-(2-chloro-4-acetamido-benzoylamino)-3-cyclohexyl]-p-
yrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00365##
[0366] Example II-51
(S,S,S)-(3S)-({1-[(2S)-(2-chloro-4-methoxycarbonylamino-benzoylamino)-3,3--
dimethyl-butyryl]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric
acid
##STR00366##
[0367] Example II-52
(S,S,S)-(3S)-({1-[(2S)-(2-chloro-3-phenoxy-benzoylamino)-3,3-dimethyl-buty-
ryl]-pyrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00367##
[0368] Example II-53
(S,S,S)-(3S)-({1-[(2S)-(2-chloro-6-amino-benzoylamino)-3-methyl-butyryl]-p-
yrrolidine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00368##
[0369] Example II-54
(S,S,S)-(3S)-({1-[(2S)-(2-chloro-benzoylamino)-3,3-dimethyl-butyryl]-piper-
idine-(2S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00369##
[0370] Example I-55
(3S)-({2-[(2S)-(3-Methoxy-2-methyl-benzoylamino)-3,3-dimethyl-butyryl]-2-(-
1S,4R)-aza-bicyclo[2.2.1]heptane-(3S)-carbonyl}-amino)-4-oxo-butyric
acid
##STR00370##
[0371] Example II-56
(3S)-({2-[(2S)-(2-Chloro-benzoylamino)-3,3-dimethyl-butyryl]-2-(1S,4R)-aza-
-bicyclo[2.2.1]heptane-(3S)-carbonyl}-amino)-4-oxo-butyric acid
##STR00371##
[0372] Example II-57
(3S)-({2-[(2S)-(4-Acetylamino-2-chloro-benzoylamino)-3,3-dimethyl-butyryl]-
-2-(1S,4R)-aza-bicyclo[2.2.1]heptane-(3S)-carbonyl}-amino)-4-oxo-butyric
acid
##STR00372##
[0373] Example II-58
(3S)-({2-[(2S)-(2-Chloro-4-propionylamino-benzoylamino)-3,3-dimethyl-butyr-
yl]-2-(1S,4R)-aza-bicyclo[2.2.1]heptane-(3S)-carbonyl}-amino)-4-oxo-butyri-
c acid
##STR00373##
[0374] Example II-59
(3S)-({2-[(2S)-(2-Chloro-3-isobutyrylamino-benzoylamino)-3,3-dimethyl-buty-
ryl]-2-(1S,4R)-aza-bicyclo[2.2.1]heptane-(3S)-carbonyl}-amino)-4-oxo-butyr-
ic acid
##STR00374##
[0375] Example II-60
(3S)-({2-[(2S)-(2-Fluoro-3-methoxy-benzoylamino)-3,3-dimethyl-butyryl]-2-(-
1S,4R)-aza-bicyclo[2.2.1]heptane-(3S)-carbonyl}-amino)-4-oxo-butyric
acid
##STR00375##
[0376] Example II-61
(3S)-({2-[(2S)-(2-Fluoro-3-methoxy-benzoylamino)-3-methyl-butyryl]-2-(1S,4-
R)-aza-bicyclo[2.2.1]heptane-(3S)-carbonyl}-amino)-4-oxo-butyric
acid
##STR00376##
[0377] Example II-62
(3S)-({2-[(2S)-(4-Acetylamino-3-chloro-benzoylamino)-3,3-dimethyl-butyryl]-
-2-(1S,4R)-aza-bicyclo[2.2.1]heptane-(3S)-carbonyl}-amino)-4-oxo-butyric
acid
##STR00377##
[0378] Example II-63
(3S)-({2-[(2S)-(3-Chloro-4-propionylamino-benzoylamino)-3,3-dimethyl-butyr-
yl]-2-(1S,4R)-aza-bicyclo[2.2.1]heptane-(3S)-carbonyl}-amino)-4-oxo-butyri-
c acid
##STR00378##
[0379] Example II-64
(3S)-({2-[(2S)-(Isoquinolin-1-ylcarbonylamino)-3,3-dimethyl-butyryl]-2-(1S-
,4R)-aza-bicyclo[2.2.1]heptane-(3S)-carbonyl}-amino)-4-oxo-butyric
acid
##STR00379##
[0380] Example II-65
(3S)-({2-[(2S)-(4-Amino-3-chloro-benzoylamino)-3,3-dimethyl-butyryl]-2-(1S-
,4R)-aza-bicyclo[2.2.1]heptane-(3S)-carbonyl}-amino)-4-oxo-butyric
acid
##STR00380##
[0381] Example II-66
(3S)-({2-[(2S)-(4-Amino-3-chloro-benzoylamino)-3-methyl-butyryl]-2-(1S,4R)-
-aza-bicyclo[2.2.1]heptane-(3S)-carbonyl}-amino)-4-oxo-butyric
acid
##STR00381##
[0383] The characterization data for compounds II-1 to II-66 is
summarized in Table 4 below and includes HPLC, LC/MS (observed) and
.sup.1H NMR data. .sup.1H NMR data was obtained at 400 MHz, and was
found to be consistent with structure.
TABLE-US-00004 TABLE 4 Characterization Data for Selected Compounds
of Formula II (According to Compound Number) M + 1 No. (obs)
.sup.1H-NMR II-1 462.1 (DMSO-d.sub.6) 0.82-0.98 (6H, m), 1.89-2.07
(5H, m), 2.10 (3H, s), 3.0 (1H, m), 3.63 (1H, m), 3.79 (3H, s),
3.88 (1H, m), 4.00 (1H, m), 4.25 (1H, m), 4.40-4.44 (2H, m), 5.45
(1H, br s), 6.83 (1H, d), 7.00 (1H, d), 7.19 (1H, t), 7.77 (1H, br
s), 8.32-8.50 (2H, m) II-2 452.0 (DMSO-d.sub.6) 0.95-0.99 (6H, m),
1.87-2.09 (5H, m), 3.00 (1H, m), 3.64 (1H, m), 3.85 (1H, m), 4.04
(1H, m), 4.25 (1H, m), 4.40 (1H, m), 4.47 (1H, m), 5.45 (1H, m),
7.34-7.49 (4H, m), 7.78 (1H, m), 8.40 (1H, m), 8.64 (1H, m) II-3
432.1 (DMSO-d.sub.6) 0.94-0.99 (6H, m), 1.87-2.09 (5H, m), 2.30
(3H, s), 2.90 (1H, m), 3.64 (1H, m), 3.88 (1H, m), 4.03 (1H, m),
4.30 (1H, m), 4.44 (1H, m), 5.45 (1H, m), 7.19-7.33 (4H, m), 7.77
(1H, br s), 8.35-8.40 (2H, m) II-4 448.0 (CD.sub.3OD) 1.05-1.18
(6H, m), 2.00-2.30 (5H, m), 2.52-2.75 (2H, m), 3.66-3.83 (1H, m),
3.92-4.03 (1H, m), 4.26-4.35 (1H, m), 4.45-4.55 (1H, m), 4.61-4.70
(1H, m), 4.78-4.85 (1H, m), 7.13 (1H, t), 7.21 (1H, d), 7.57 (1H,
t), 8.00 (1H, d), 8.69 (1H, d) II-5 502.0 (CD.sub.3OD) 0.98-1.15
(6H, m), 1.95-2.26 (5H, m), 2.54-2.76 (2H, m), 3.73-3.84 (1H, m),
3.99-4.06 (1H, m), 4.21-4.32 (1H, m), 4.45-4.53 (1H, m), 4.60-4.70
(2H, m), 7.30-4.47 (2H, m), 7.54-7.64 (2H, m) II-6 448.1
(DMSO-d.sub.6) 0.92-0.98 (6H, m), 1.85-2.04 (5H, m), 2.07 (3H, s),
3.00 (1H, m), 3.63 (1H, m), 3.87 (1H, m), 4.03 (1H, m), 4.25 (1H,
m), 4.41 (1H, m), 5.45 (1H, m), 6.68 (1H, m), 6.82 (1H, m), 7.01
(1H, m), 7.81 (1H, m), 8.25 (1H, d), 8.40 (1H, m), 9.5 (1H, m) II-7
447.0 (CD.sub.3OD) 1.02-1.18 (6H, m), 1.88-2.28 (5H, m), 2.39 (3H,
s), 2.50-2.78 (2H, m), 3.75-3.83 (1H, m), 4.00-4.10 (1H, m),
4.21-4.32 (1H, m), 4.45-4.52 (1H, m), 4.60-4.65 (2H, m), 7.39-7.54
(3H, m) II-8 446.0 (DMSO-d.sub.6) 0.94-0.99 (6H, m), 1.71-2.12 (4H,
m), 2.33 (1H, br s), 2.67 (1H, br s), 2.94-3.07 (1H, m), 3.61-3.69
(1H, m), 3.82-3.87 (1H, m), 4.03-4.10 (1H, m), 4.19-4.28 (1H, m),
4.30-4.43 (2H, m), 5.42-5.47 (1H, m), 7.28-7.30 (1H, m), 7.37-7.40
(1H, m), 7.68-7.82 (2H, m), 8.77 (1H, d) II-9 519.9 (DMSO-d.sub.6)
0.94-0.99 (6H, m), 1.86-2.09 (5H, m), 3.00 (1H, m), 3.65 (1H, m),
3.84 (1H, m), 4.05 (1H, m), 4.24 (1H, m), 4.40 (1H, m), 4.51 (1H,
m), 5.45 (1H, m), 7.57-7.62 (2H, m), 7.77 (1H, d), 7.90 (1H, m),
8.40 (1H, d), 8.87 (1H, d) II-10 466.0 (DMSO-d.sub.6) 0.93-0.99
(6H, 2 .times. d), 1.77-2.19 (5H, m), 2.29 (3H, s), 2.97 (1H, br
s), 3.62-3.65 (1H, m), 3.85-3.88 (1H, m), 4.00-4.32 (2H, br m),
4.41-4.53 (2H, m), 5.45 (1h, br s), 7.18-7.27 (2H, m), 7.45-7.50
(1H, m), 7.85 (1h, br d), 8.41 (1H, br d), 8.57 (1H, d) II-11 485.9
(DMSO-d.sub.6) 0.82-0.86 (3H, m), 0.93-0.98 (3H, m), 1.87-2.08 (5H,
m), 3.00 (1H, m), 3.64 (1H, m), 3.82 (1H, m), 4.10 (1H, m), 4.30
(1H, m), 4.45 (1H, m), 4.47 (1H, m), 5.44 (1H, d), 7.37 (1H, m),
7.47 (1H, m), 7.65 (1H, m), 7.77 (1H, m), 8.40 (1H, m), 8.72 (1H,
m) II-12 485.9 (DMSO-d.sub.6) 0.94-0.99 (6H, m), 1.91-2.09 (5H, m),
3.00 (1H, m), 3.64 (1H, m), 3.83 (1H, m), 4.03 (1H, m), 4.20 (1H,
m), 4.40 (1H, m), 4.47 (1H, m), 5.45 (1H, m), 7.37 (1H, s),
7.50-7.52 (2H, m), 7.78 (1H, m), 8.44 (1H, m), 8.79 (1H, m) II-13
486.3 (DMSO-d.sub.6) 0.82-0.86 (3H, m), 0.92-0.99 (3H, m),
1.80-1.87 (2H, m), 1.99-2.02 (4H, m), 2.48 (0.5H, m), 2.95 (0.5H,
m), 3.51 (1H, m), 3.80-4.56 (4H, m), 5.00 and 5.47 (1H, 2 .times.
m), 7.37-7.48 (3H, m), 7.76-8.32 (1H, m), 8.95-9.39 (1H, 3 .times.
dd) II-14 446.0 (DMSO-d.sub.6) 0.93-0.99 (6H, m), 1.80-2.09 (5H,
m), 2.17 (6H, d), 2.95 (1H, br s), 3.63-3.65 (1H, m), 3.96-3.99
(1H, m), 4.10 (1H, br s), 4.30 (1H, br s), 4.44 (1H, t), 5.48 (1H,
br s), 7.00 (2H, d), 7.14 (1H, t), 7.78 (1H, br s), 8.50 (1H, br
s), 8.55 (1H, d) II-15 433.1 (DMSO-d.sub.6) 0.91-1.02 (6H, m),
1.80-2.20 (5H, m), 2.66-2.68 (3H, s), 3.00 (1H, m), 3.62-3.85 (3H,
m), 4.10 (1H, m), 4.24 (1H, m), 4.51 (1H, m), 5.72 (1H, m),
7.73-7.76 (2H, m), 8.19 (1H, m), 8.52 (1H, m), 8.75 (1H, d), 8.90
(1H, m) II-16 433.1 (DMSO-d.sub.6) 0.9-1.05 (6H, m), 1.8-2.2 (6H,
m), 2.3-2.4 (1H, m), 2.7-2.75 (1H, m), 2.9-3.0 (1H, m), 3.65-3.75
(1H, m), 3.8-3.9 (1H, m), 4.1-4.15 (1H, m), 4.3-4.4 (1H, m),
4.45-4.65 (1H, m), 7.8-7.9 (1H, m), 8.7-8.8 (2H, d), 8.9.8.95 (1H,
m) II-17 438.0 (DMSO-d.sub.6) 0.83-0.99 (6H, m), 1.80-2.20 (5H, m),
2.40 (3H, s), 3.00 (1H, m), 3.61 (1H, m), 3.81 (1H, m), 4.10 (1H,
m), 4.25 (1H, m), 4.42-4.46 (2H, m), 5.44 (1H, br s), 6.97 (1H, m),
7.34 (1H, m), 7.59 (1H, m), 7.81 (1H, m), 8.49 (1H, m) II-18 487.0
(DMSO-d.sub.6) 0.92-1.00 (6H, m), 1.75-2.08 (5H, m), 2.30-2.34 (1H,
m), 2.99 (1H, dd), 3.62-3.67 (1H, m), 3.78-3.82 (1H, m), 3.78-3.82
(1H, m), 4.05-4.26 (1H, m), 4.38-4.54 (2H, m), 5.44-5.72 (1H, m),
7.37-7.41 (1H, m), 8.41-8.43 (2H, m), 8.97-9.00 (1H, d) II-19 487.0
(DMSO-d.sub.6) 0.94-1.00 (6H, m), 1.77-2.15 (5H, m), 3.02 (1H, dd),
3.61-3.70 (1H, m), 3.80-3.90 (1H, m), 4.03-4.08 (1H, m), 4.52-4.56
(1H, m), 4.95 (2H, br s), 5.45 (1H, s), 8.42 (1H, d), 8.67 (2H, s),
9.17 (1H, d) II-20 476.4 (DMSO-d.sub.6) 0.91-1.11 (9H, m),
1.70-2.14 (7H, m), 2.31 (1H, m), 3.01 (1H, m), 3.50-3.97 (5H, m),
4.00-4.62 (3H, m), 5.50 (1H, m), 6.77 (1H, d), 7.00 (1H, d), 7.18
(1H, dd), 7.50-8.50 (3H, m) II-21 502.1 (DMSO-d.sub.6) 1.80-2.00
(3H, m), 2.11 (4H, overlapping s and m), 2.60-2.80 (2H, m),
3.64-3.69 (1H, m), 3.80 (3H, s), 4.10 (1H, vbrs), 4.30 (1H, vbrs),
5.00 (1H, m), 6.86 (1H, d), 7.03 (1H, d), 7.22 (1H, t), 8.45 (1H,
vbrs), 8.81 (1H, d) II-22 462.4 (DMSO-d.sub.6) 0.93-1.00 (6H, m),
1.70-2.15 (5H, m), 2.22 (3H, s), 2.33 (1H, d), 2.99 (1H, dd),
3.60-3.65 (2H, m), 3.74 (3H, s), 4.04-4.08 (1H, m), 4.21-4.27 (1H,
m), 4.40-4.58 (2H, m), 5.46 (1H, brd d), 6.78-6.81 (1H, m),
6.85-6.91 (1H, m), 7.09-7.14 (1H, m), 8.37 (2H, 2 .times. brd d)
II-23 517.0 (DMSO-d.sub.6) 1.77-2.19 (5H, m), 2.95-3.28 (3H, m),
3.60 (1H, brd d), 3.71-3.78 (4H, m), 4.10-4.42 (6H, m), 4.97 (1H,
brd s), 5.45-72 (1H, m), 6.74 (1H, d), 6.97 (1H, d), 7.10-7.22 (1H,
m), 7.44 (1H, m), 8.37-8.68 (2H, m), 9.05 (1H, brd s) II-24 492.0
(DMSO-d.sub.6) 1.75-1.98 (3H, m), 2.08-2.13 (1H, m), 2.64-2.77 (2H,
m), 2.99 (0.5H, dd), 3.63-3.73 (2H, m), 4.08 (0.5H, brt), 4.20
(0.5H, dd), 4.23-4.49 (3 multiplets, 1H total), 5.00-5.10 (1H, m),
5.42 (0.5H, s), 7.36-7.52 (4H, m), 7.77 (1H, m), 8.30 (0.5H, d),
9.09 (1H, d) II-25 507.0 (DMSO-d.sub.6) 1.79-1.96 (5H, m),
2.94-3.28 (3H, m), 3.58 (1H, brd d), 3.73 (1H, brd d), 4.04-4.59
(2H, m), 4.98-5.02 (1h, m), 5.54-5.74 (2H, m), 7.26-7.46 (5H, m),
8.43 (1H, d), 8.82 (1H, d), 9.39 (1H, brd s) II-26 446.6
(DMSO-d.sub.6) 1.05 (9H, s), 1.15 (3H, t), 1.8-2.1 (4H, m), 2.3
(3H, s), 2.4-2.5 (1H, m), 2.9-3.0 (1H, m), 3.7-3.75 (1H, m),
3.8-3.85 (1H, m), 4.1-4.15 (0.5H, m), 4.25-4.3 (1H, m), 4.4-4.5
(0.5H, m), 4.7-4.75 (1H, m), 5.55-5.6 (1H, m), 7.2-7.4 (4H, m),
7.7-7.75 (1H, m), 8.1-8.15 (1H, m), 8.35-8.4 (1H, m) II-27 486.5
(DMSO-d.sub.6) 0.95-1.05 (6H, m), 1.8-2.1 (4H, m), 2.4-2.5 (1H, m),
3.0-3.1 (1H, m), 3.7-3.75 (1H, m), 3.8-3.85 (1H, m), 4.1-4.15
(0.5H, m), 4.25-4.3 (1H, m), 4.4-4.5 (0.5H, m), 5.55-5.6 (1H, m),
7.4-7.45 (1H, m), 7.6-7.8 (3H, m), 8.4-8.45 (1H, m), 8.75-8.8 (1H,
m) II-28 466.1 (CDCl.sub.3) 1.11-1.16 (9H, m), 1.94-2.22 (4H, m),
2.38-2.50 (2H, m), 2.77-2.87 (1H, m), 3.71-3.79 (1H, m), 3.96-4.06
(1H, m), 4.56-4.67 (2H, m), 4.85-4.91 (1H, m), 6.99-7.02 (1H, m),
7.28-7.45 (3H, m), 7.60-7.84 (2H, m) II-29 500.2 (CDCl.sub.3) 1.07
(9H, s), 1.85-2.19 (2H, m), 2.37-2.40 (2H, m), 2.81-3.07 (1H, m),
3.37 (1H, brs), 4.01 (1H, brs), 4.46-4.67 (2H, m), 4.87 (1H, d),
5.73 (1H, brs), 6.68 (1H, brs), 7.38-7.74 (5H, m) II-30 496.2
(CD.sub.3OD) 1.15 (9H, s), 1.85-2.20 (4H, m), 2.46-2.72 (2H, m),
3.74-3.81 (1H, m), 3.92 (3H, s), 3.93-4.03 (1H, m), 4.20-4.31 (1H,
m), 4.45-4.52 (1H, m), 4.60-4.75 (1H, m), 4.83 (1H, s), 7.00 (1H,
d), 7.15 (1H, d), 7.33 (1H, t) II-31 480.5 (DMSO-d.sub.6) 1.05 (9H,
s), 1.8-2.1 (4H, m), 2.4-2.5 (1H, m), 3.75-3.8 (1H, m), 3.8-3.85
(1H, m), 3.9 (3H, s), 4.1-4.3 (1H, m), 4.7 (1H, d), 5.3-5.5 (0.5H,
br s), 7.1-7.3 (3H, m), 7.7-7.8 (1H, m), 8.0-8.1 (1H, m), 8.35-8.45
(1H, m) II-32 550.3 (DMSO-d.sub.6) 0.91-1.10 (9H, m), 1.70-2.15
(5H, m), 2.60-3.08 (1H, m), 3.60-3.90 (2H, m), 3.98-4.71 (3H, m),
5.40-5.80 (1H, m), 7.30-7.91 (3H, m), 8.30-8.80 (3H, m) II-33 523.3
(DMSO) 0.60-0.90 (4H, m, cyclopropyl CH2), 0.92-1.10 (9H, m, tBu),
1.71-2.21 (5H, m, CH2), 2.65-3.10 (1H, brm, CH2), 3.36-3.50 (1H, m,
CH), 3.60-4.75 (6H, m, CH), 6.92 (1H, d, aryl H), 7.36 (1H, m, aryl
H), 7.45 (1H, m, aryl H), 7.65-8.60 (3H, m, NH, OH) II-34 480.3
(DMSO) 0.99-1.10 (9H, m, tBu), 1.70-2.12 (5H, m, CH2), 2.35 (3H, s,
CH3), 2.60-3.08 (1H, m, CH2), 3.58-3.87 (2H, m, CH), 4.00-4.70 (3H,
m, CH), 5.38-5.79 (1H, m, CH), 7.12 (1H, d, aryl H), 7.24 (1H, m,
aryl H), 7.38 (1H, m, aryl H), 7.69-8.55 (3H, m, NH, OH) II-35 482
CD3OD 1.01-1.15 (6H, m), 1.95-2.22 (5H, m), 2.48-2.69 (2H, m),
3.73-3.80 (1H, m), 4.92 (3H, s), 3.99-4.19 (1H, m), 4.20-4.30 (1H,
m), 4.58-4.67 (2H, m), 7.00 (1H, d), 7.14 (1H, d), 7.31 (1H, t)
II-36 494.4 (DMSO) 0.94-1.08 (9H, s, tBu), 1.19 (3H, t, CH3),
1.70-2.40 (5H, m, CH2), 2.60-3.08 (3H, m, CH2), 3.69 (1H, m, CH),
3.81 (1H, m, CH), 4.04-4.71 (3H, m, CH), 5.40-5.80 (1H, m, CH),
7.14 (1H, m, aryl H), 7.31 (1H, m, aryl H), 7.39 (1H, m, aryl H),
7.70-8.50 (3H, m, NH, OH)
II-37 482.5 (DMSO) 0.9-1.0 (6H, m), 1.85-2.3 (4H, m), 3.0-3.1 (1H,
m), 3.65-3.7 (1H, m), 3.78 (3H, s), 3.8-3.85 (1H, m), 4.1-4.15
(0.5H, m), 4.25-4.3 (0.5H, m), 4.5-4.55 (1H, m), 5.5-5.55 (1H, m),
6.93 (1H, d), 6.98 (1H, s), 7.35 (1H, d), 7.75-7.8 (1H, m), 8.45
(1H, d) II-38 536 (CD3OD) 0.34-0.40 (2H, m), 0.60-0.67 (2H, m),
1.16 (9H, s), 1.25-1.32 (1H, m), 1.93-2.22 (4H, m), 2.50-2.66 (2H,
m), 3.74-3.84 (1H, m), 3.91-4.03 (3H, m), 4.22-4.32 (1H, m),
4.45-4.54 (1H, m), 4.61-4.69 (1H, m), 4.82 (1H, d), 6.99 (1H, d),
7.12 (1H, d), 7.32 (1H, t), 8.40 (1H, d) II-39 482 (CD3OD) 1.12
(9H, s), 1.90-2.22 (4H, m), 2.512.70 (2H, m), 3.75-3.83 (1H, m),
3.97-4.05 (1H, m), 4.23-4.30 (1H, m), 4.46-4.54 (1H, m), 4.63-4.70
(1H, m), 4.83 (1H, d), 6.91 (1H, d), 6.99 (1H, d), 7.17 (1H, t),
8.36 (1H, d) II-40 509.3 (DMSO) 0.93-0.98 (6H, m) 1.71-2.09 (10H,
m), 2.35-2.45 (2H, m), 3.61-3.64 (1H, m), 4.02-4.04 (1H, m),
4.06-4.35 (2H, m), 4.43-4.46 (1H, m), 7.33 (1H, d), 7.43-7.46 (1H,
m), 7.80 (1H, brd s), 8.28-8.49 (2H, m), 10.25 (1H, brd s) II-41
523.3 (DMSO) 0.95-1.08 (9H, s, tBu), 1.70-2.38 (8H, m, COCH3, CH2),
2.58-3.08 (1H, m, CH2), 3.65 (1H, m, CH), 3.82 (1H, m, CH0),
3.95-4.69 (3H, m, CH), 5.40-5.60 (1H, m, CH), 7.09 (1H, m, aryl H),
7.31 (1H, m, aryl H), 7.64-8.60 (4H, m, aryl H, NH), 9.55 (1H, m,
CH) II-42 503.4 (DMSO) 0.91-1.08 (9H, s, tBu), 1.70-2.40 (11H, m,
CH3, COCH3, CH2), 2.60-3.08 (1H, m, CH2), 3.66 (1H, m, CH), 3.87
(1H, m, CH), 4.00-4.65 (3H, m, CH), 5.40-5.78 (1H, m, CH), 7.04
(1H, m, aryl H), 7.18 (1H, m, aryl H), 7.38 (1H, m, aryl H),
7.65-7.88 (1H, m, NH), 8.07-8.70 (2H, m, NH), 9.34 (1H, m, CH)
II-43 523.3 (DMSO) 1.03 (9H, s), 1.71-2.00 (3H, m), 2.07 (3H, s),
2.55-2.73 (1H, m), 2.97 (1H, dd), 3.60-3.67 (1H, m), 3.75-3.82 (1H,
m), 3.98-4.04 (1H, m), 4.19-4.24 (1H, m), 4.37-4.45 (1H, m), 4.63
(1H, d), 5.45 (1H, d), 7.33-7.35 (1H, m), 7.43-7.45 (1H, d),
7.76-7.83 (2H, m), 8.25-8.28 (1H, m), 8.41-8.58 (1H, m), 10.27 (1H,
s) II-44 507.4 (DMSO) 1.01 (9H, 2 .times. s), 1.72-1.99 (4H, m),
2.05-2.09 (4H, m), 2.35-2.57 (2H, m), 2.71-3.00 (1H, brd m),
3.60-3.65 (1H, m), 3.71-3.80 (1H, m), 4.08-4.37 (2H, brd m), 4.70
(1H, d), 7.32 (1H, dd), 7.65-7.80 (3H, m), 8.33-8.52 (1H, brd m),
10.37 (1H, s) II-45 493.4 (DMSO) 0.94 (6H, dd), 1.72-1.99 (10H, m),
2.36-2.52 (2H, m), 3.57-3.68 (1H, m), 3.76-3.88 (1H, m), 4.20-4.43
(2H, m), 4.51-4.55 (1H, m), 7.30 (1H, dd), 7.58-7.77 (3H, m),
8.00-8.04 (1H, m), 10.34 (1H, s) II-46 510.5 (DMSO) 0.95-1.0 (6H,
m), 1.25 (6H, d), 1.85-2.2 (4H, m), 3.0-3.1 (1H, m), 3.9-4.0 (3H,
m), 4.2-4.3 (0.5H, m), 4.4-4.5 (0.5H, m), 4.7-4.8 (1H, m), 6.9-6.95
(1H, d), 6.99 (1H, s), 7.3 (1H, d), 8.3-8.4 (1H, m) II-47 482.5
(DMSO) 1.05 (9H, m), 1.8-2.1 (4H, m), 2.6-2.7 (1H, m), 2.9-3.0 (2H,
m), 3.6-3.7 (2H, m), 3.8-3.9 (1H, m), 4.0-4.1 (1H, m), 4.2-4.3 (1H,
m), 4.6-4.65 (1H, m), 5.5-5.55 (1H, m), 6.75-6.85 (2H, m), 7.35
(1H, d), 7.75 (1H, d), 8.0-8.1 (1H, m), 8.35 (1H, m), 10.25 (1H, s)
II-48 510.5 (DMSO) 1.03 (9H, s), 1.80-2.10 (4H, m), 3.00 (1H, br
s), 3.30 (3H, s), 3.66 (1H, m), 3.81 (1H, m), 4.06 (1H, m), 4.25
(1H, m), 4.44 (2H, s), 4.65 (1H, d), 5.46 1H, br s), 7.29-7.39 (3H,
m), 7.77 (1H, br s), 8.43 (1H, m) II-49 551.5 (DMSO) 1.03 (9H, s),
1.09 (3H, m), 1.11 (3H, m), 1.79-2.15 (4H, m), 2.32 (1H, m), 2.98
(1H, m), 3.51 (1H, m), 3.79 (1H, m), 4.10 (1H, m), 4.23 (1H, m),
4.40-4.65 (2H, m), 5.45-5.73 (1H, m), 7.35 (1H, m), 7.49 (1H, m),
7.76-7.84 (2H, m), 8.23-8.60 (2H, m), 10.11 (1H, s) II-50 493.3
(DMSO) 0.92-1.19 (4H, m), 1.49-1.90 (9H, m), 1.91-1.99 (2H, m),
2.06 (4H, brd s), 2.49-2.52 (2H, m), 3.57-3.68 (1H, m), 3.80-3.90
(1H, m), 4.01-4.28 (2H, m), 4.46 (1H, t), 7.32 (1H, d), 7.43 (1H,
dd), 7.81 (2H, brd s), 8.31-8.78 (1H, m), 8.46 (1H, d), 10.22 (1H,
s) II-51 539.3 (DMSO) 0.90-1.07 (9H, s, tBu), 1.70-2.40 (4H, brm,
CH2), 2.54-3.07 (1H, m, CH2), 3.52-3.88 (5H, m, CH3, CH), 4.00-4.65
(3H, m, CH), 5.40-5.80 (1H, m, CH), 7.30-7.44 (2H, m, aryl H), 7.60
(1H, m, aryl H), 7.67 (1H, br, NH), 8.10-8.70 (2H, m, NH), 10.00
(1H, m, CH) II-52 558.3 (DMSO) 0.91-1.11 (9H, s, tBu), 1.70-2.41
(4H, m, CH2), 2.56-3.09 (1H, m, CH2), 3.60-3.90 (2H, m, CH),
4.14-4.72 (3H, m, CH), 5.38-5.80 (1H, m, CH), 6.98 (2H, m, aryl H),
7.07-7.20 (3H, m, aryl H), 7.31-7.46 (3H, m, aryl H), 7.66-8.67
(3H, m, NH, OH) II-53 467 (DMSO) 0.83-1.04 (6H, m), 1.81-2.08 (5H,
m), 3.34-3.63 (1H, m), 3.84-3.90 (1H, m), 4.00-4.60 (3H, m),
5.29-5.75 (2H, m), 6.53-6.59 (1H, m), 6.70-6.90 (1H, m), 7.20-7.35
(0.5H, m), 7.78 (0.5H. brs), 8.43-8.60 (2H, m) II-55 502.6 (DMSO)
0.96 (1H, s). 1.03 (9H, s), 1.30-1.39 (2H, m), 1.68-1.71 (2H, m),
1.79-1.82 (1H, m), 1.97 (1H, brd), 2.11 (3H, s), 3.79 (3H, s), 3.84
(1H, vbrs), 4.09 (1H, vbrs), 4.56-4.58 (1H, m), 4.67 (1H, d), 6.81
(1H, d), 7.00 (1H, d), 7.19 (1H, t), 7.79 (0.5H, vbrs), 7.93 (1H,
brd), 8.42 (0.5H, vbrs) II-56 492.5 (CDCl3) 1.08-1.14 (9H, m),
1.85-2.05 (4H, m), 2.32-2.45 (1H, m), 2.79-2.85 (1H, m), 3.01-3.07
(1H, m), 4.13-4.17 (1H, m), 4.53-4.70 (1H, m), 4.98 (1H, t), 5.70
and 5.81 (1H total, brs and brd), 6.91-7.00 (1H, m), 7.34-7.44 (3H,
m), 7.67-7.75 (1H, m) II-57 549.5 (DMSO) 1.03 (9H, s), 1.31-1.38
(2H, m), 1.62-1.74 (3H, m), 1.98 (1H, brt), 2.07 (3H, s), 2.36 (1H,
vbrs), 2.83 (1H, vbrs), 3.84 (1H, brs), 4.17 (1H, vbrs), 4.54-4.57
(1H, m), 4.70 (1H, d), 7.34 (1H, d), 7.42-7.45 (1H, m), 8.16 (1H,
t), 8.37 (1H, brs), 10.23 (1H, s) II-58 563.5 (CD3OD) 1.17 (9H, s),
1.21 (3H, t), 1.41-1.55 (2H, m), 1.75-1.90 (3H, m), 2.03-2.19 (1H,
m), 2.37-2.50 (3H, m), 2.58-2.78 (2H, m), 3.87-4.02 (1H, m),
4.20-4.30 (1H, m), 4.55-4.70 (2H, m), 4.91 (1H, obscured), 7.45
(1H, d), 7.51 (1H, d), 7.85 (1H, s), 8.29 (1H, d) II-59 577.5
(DMSO) 1.05 (9H, s), 1.15 (6H, d), 1.35-1.5 (2H, m), 1.75-1.9 (3H,
m), 2.0-2.1 (1H, m), 2.3-2.45 (1H, m), 2.7-2.9 (1H, m), 4.05-4.15
(1H, m), 4.65 (1H, s), 4.7-4.75 (1H, m), 7.15 (1H, d), 7.35 (1H,
t), 7.7 (1H, d), 8.4-8.55 (2H, m), 9.5 (1H, s) II-60 506.5 (DMSO)
1.03 (9H, s), 1.31-1.38 (2H, m), 1.68 (3H, m), 2.30-2.33 (2H, m),
2.67 (0.5H, brs), 2.99 (0.5H, brs), 3.34 (0.5H, brs), 3.76 (3H, s),
4.04 (0.5H, m), 4.58 (1H, s), 4.72 (1H, d), 7.09-7.12 (1H, m),
7.16-7.20 (1H, m), 7.26-7.30 (1H, m), 7.78 (0, 5H, vbrs), 8.02 (1H,
brs), 8.42 (0.5H, vbrs) II-61 492.8 (DMSO) 0.95 (3H, d), 0.10 (3H,
d), 1.17 (1H, m), 1.32 (1H, m), 1.64-1.80 (3H, m), 2.00 (1H, m),
2.30 (1H, br s), 2.67 (0.5H, br s), 2.99 (0.5H, br s), 3.75 (0.5H,
br s), 3.85 (3H, s), 4.06 (0.5H, m), 4.50-4.55 (2H, m), 5.42 (1H,
br s), 7.07 (1H, m), 7.17 (1H, m), 7.26 (1H, m), 7.80 (1H, br s),
8.35 (1H, m) II-62 549.5 DMSO) .delta. 1.04 (9H, s), 1.29-1.34 (2H,
m), 1.59-1.67 (3H, m), 1.91-1.97 (1H, m), 2.13 (3H, s), 2.96 (1H,
vbrs), 3.77 (1H, vbrs), 4.10 (1H, vbrs), 4.72 (1H, s), 4.76 (1H,
d), 7.80-7.83 (1H, m), 7.88-7.91 (1H, m), 8.00-8.02 (1H, m),
8.18-8.24 (12H, m), 8.39 (1H, vbrs), 9.62 (1H, s) II-63 563.5
(DMSO) 1.05 (9H, s), 1.09 (3H, t), 1.19-1.37 (3H, m), 1.47-1.77
(3H, m), 1.91-1.99 (1H, m), 2.28 (0.5H, brdd), 2.48 (2H, q),
2.63-2.74 (1H, m), 3.01 (0.5H, dd), 3.63 (0.5H, s), 3.78-4.37 (2H,
total, m), 4.42-4.59 (1H. m), 4.75 (1H, d), 5.42 (0.5H, d), 7.76
(0.5H, d), 7.80-7.83 (1H, m), 7.87 (1H, d), 8.01 (1H, m), 8.08-8.15
(1H, m), 8.36 (0.5H, d), 9.53 (1H, s) II-64 509.5 (DMSO) 1.07 (9H,
s), 1.34-1.37 (2H, m), 1.64-1.72 (3H, m), 1.95-2.04 (1H, m),
2.31-2.35 (1H, m), 2.65-2.70 (1H, m), 3.01-3.03 (1H, m), 3.99
(0.5H, m), 4.26-4.28 (0.5H, m), 4.68 (1H, s), 4.82 (1H, d), 5.45
(0.5H, s), 7.73-7.86 (3H, m), 8.05-8.08 (2H, m), 8.49 (0.5H, d),
8.57-8.59 (1H, m), 8.69 (0.5H, d), 9.15 (1H, d) II-65 507.5 (DMSO)
1.02 (9H, s), 1.28-1.34 (2H, m), 1.57-1.64 (3H, m), 1.90-1.96 (1H,
m), 3.72-3.80 (1H, m), 4.50 (1H, brs), 4.72-4.74 (1H, m), 5.91 (1H,
s), 6.76 (1H, d), 7.58-7.61 (1H, m), 7.81-7.83 (1H, m) II-66 493.5
/
Example III
Biological Methods
[0384] Compounds of this invention may be tested using the methods
described below. Table 5 lists caspase-1 and caspase-8 enzyme
inhibition data for compounds II-1-II-25. In the Table, compounds
with a Ki of <10 are assigned category A, compounds with a Ki of
10-20 are assigned category B, and compounds with a Ki of 21-30 are
assigned category C.
In Vitro Assays
Enzyme Inhibition
[0385] Ki values for test compounds with caspase-1 and caspase-8
were obtained by the method of Margolin et al. (J. Biol. Chem., 272
pp. 7223-7228 (1997)). Other caspases may be assayed similarly
(see, e.g., WO 99/47545). Assays were performed in 10 mM Tris
(Sigma Corp, St Louis Mo.) pH 7.5, 1 mM Dithiothreitol (DTT,
Research Organic INC, Cleveland, Ohio) and 0.1% CHAPS (Pierce,
Rockford Ill.) at 37.degree. C. For caspase-3, a solution of 8%
glycerol was added to the assay buffer to improve enzyme stability.
A 65 .mu.L aliquot of the assay buffer and 5 .mu.L aliquot of the
appropriate dilutions of inhibitor in DMSO where pipetted into a 96
well plate, treated with 10 .mu.L of caspase, then diluted in assay
buffer (0.5-40 nM active protein by active site titration). A
control containing DMSO but no compound was included for each
determination. The plates were then incubated for 15 minutes at
37.degree. C., before addition of the appropriate substrate (20
.mu.L, final concentration 1-4.times.K.sub.M, final assay volume
100 .mu.L) to initiate the reaction. Reaction rates were measured
at 37.degree. C. either by following the time dependant increase in
absorbance at 405 nM (for the pNA substrates) or in fluorescence
(Ex 390, Em 460) (for the AMC substrates). The rates obtained were
plotted against inhibitor concentration and the data fit to the
Morrison tight-binding equation for competitive inhibitors
(Morrison, J. F., Biochem. Biophys. Acta, 185 pp. 269-286 (1969)).
The substrates used for the individual assays were as follows:
[0386] Caspase-1 Suc-YVAD-pNA (Bachem, King of Prussia, Pa.) (final
concentration in the assay 80 .mu.M);
[0387] Caspase-8 Ac-DEVD-pNA (Bachem, King of Prussia, Pa.) (final
concentration in assay 80 .mu.M).
TABLE-US-00005 TABLE 5 Caspase-1 (C1) and caspase-8 (c8) inhibition
data. Ki C1 Ki C8 Compound (nM) (nM) II-1 A A II-2 A A II-3 A A
II-4 A B II-5 A B II-6 A A II-7 A B II-8 A B II-9 A B II-10 A B
II-11 A C II-12 A B II-13 B B II-14 B A II-15 A C II-16 A C II-17 A
A II-18 A B II-19 B A II-20 A A II-21 A C II-22 A C II-23 A C II-24
A C II-25 A C II-26 A A II-27 A A II-28 A A II-29 A A II-30 A A
II-31 A A II-32 A A II-33 A A II-34 A A II-35 A A II-36 A A II-37 A
B II-38 A A II-39 A A II-40 A B II-41 A A II-42 A B II-43 A A II-44
A A II-45 A A II-46 A C II-47 A A II-48 A A II-49 A A II-50 A C
II-51 A A II-52 A A II-53 A C II-55 A A II-56 A A II-57 A A II-58 A
A II-59 A A II-60 A A II-61 A A II-62 A B II-63 A B II-64 B B II-65
A A II-66 B A
PBMC Cell Assay
[0388] IL-1.beta. Assay with a Mixed Population of Human Peripheral
Blood Mononuclear Cells (PBMC) or Enriched Adherent Mononuclear
Cells
[0389] Processing of pre-IL-1.beta. by ICE may be measured in cell
culture using a variety of cell sources. Human PBMC obtained from
healthy donors provides a mixed population of lymphocyte subtypes
and mononuclear cells that produce a spectrum of interleukins and
cytokines in response to many classes of physiological stimulators.
Adherent mononuclear cells from PBMC provides an enriched source of
normal monocytes for selective studies of cytokine production by
activated cells.
Experimental Procedure:
[0390] An initial dilution series of test compound in DMSO or
ethanol is prepared, with a subsequent dilution into RPMI-10% FBS
media (containing 2 mM L-glutamine, 10 mM HEPES, 50 U and 50 ug/ml
pen/strep) respectively to yield drugs at 4.times. the final test
concentration containing 0.4% DMSO or 0.4% ethanol. The final
concentration of DMSO is 0.1% for all drug dilutions. A
concentration titration which brackets the apparent K.sub.i for a
test compound determined in an ICE inhibition assay is generally
used for the primary compound screen.
[0391] Generally 5-6 compound dilutions are tested and the cellular
component of the assay is performed in duplicate, with duplicate
ELISA determinations on each cell culture supernatant.
PBMC Isolation and IL-1 Assay:
[0392] Buffy coat cells isolated from one pint human blood
(yielding 40-45 ml final volume plasma plus cells) are diluted with
media to 80 ml and LeukoPREP separation tubes (Becton Dickinson)
are each overlaid with 10 ml of cell suspension. After 15 min
centrifugation at 1500-1800.times.g, the plasma/media layer is
aspirated and then the mononuclear cell layer is collected with a
Pasteur pipette and transferred to a 15 ml conical centrifuge tube
(Corning). Media is added to bring the volume to 15 ml, gently mix
the cells by inversion and centrifuge at 300.times.g for 15 min.
The PBMC pellet is resuspended in a small volume of media, the
cells are counted and adjusted to 6.times.10.sup.6 cells/ml.
[0393] For the cellular assay, 1.0 ml of the cell suspension is
added to each well of a 24-well flat bottom tissue culture plate
(Corning), 0.5 ml test compound dilution and 0.5 ml LPS solution
(Sigma #L-3012; 20 ng/ml solution prepared in complete RPMI media;
final LPS concentration 5 ng/ml). The 0.5 ml additions of test
compound and LPS are usually sufficient to mix the contents of the
wells. Three control mixtures are run per experiment, with either
LPS alone, solvent vehicle control, and/or additional media to
adjust the final culture volume to 2.0 ml. The cell cultures are
incubated for 16-18 hr at 37.degree. C. in the presence of 5%
CO.sub.2.
[0394] At the end of the incubation period, cells are harvested and
transferred to 15 ml conical centrifuge tubes. After centrifugation
for 10 min at 200.times.g, supernatants are harvested and
transferred to 1.5 ml Eppendorf tubes. It may be noted that the
cell pellet may be utilized for a biochemical evaluation of
pre-IL-1.beta. and/or mature IL-1.beta. content in cytosol extracts
by Western blotting or ELISA with pre-IL-1.beta. specific
antisera.
Isolation of Adherent Mononuclear Cells:
[0395] PBMC are isolated and prepared as described above. Media
(1.0 ml) is first added to wells followed by 0.5 ml of the PBMC
suspension. After a one hour incubation, plates are gently shaken
and nonadherent cells aspirated from each well. Wells are then
gently washed three times with 1.0 ml of media and final
resuspended in 1.0 ml media. The enrichment for adherent cells
generally yields 2.5-3.0.times.10.sup.5 cells per well. The
addition of test compounds, LPS, cell incubation conditions and
processing of supernatants proceeds as described above.
ELISA:
[0396] Quantikine kits (R&D Systems) may be used for the
measurement of mature IL-1.beta.. Assays are performed according to
the manufacturer's directions. Mature IL-1.beta. levels of about
1-3 ng/ml in both PBMC and adherent mononuclear cell positive
controls are observed. ELISA assays are performed on 1:5, 1:10 and
1:20 dilutions of supernatants from LPS-positive controls to select
the optimal dilution for supernatants in the test panel.
[0397] The inhibitory potency of the compounds can be represented
by an IC.sub.50 value, which is the concentration of inhibitor at
which 50% of mature IL-1.beta. is detected in the supernatant as
compared to the positive controls.
[0398] The skilled practitioner realizes that values obtained in
cell assays may depend on multiple factors. The values may not
necessarily represent fine quantitative results.
[0399] Selected compounds of this invention have been tested for
inhibition of IL-1.beta. release from PBMCs with IC50 values
between 300 nM and 4 .mu.M.
Whole Blood Assay for IL-1.beta. Production
[0400] Whole blood assay IC.sub.50 values for compounds of this
invention may be obtained using the method described below:
Purpose:
[0401] The whole blood assay is a simple method for measuring the
production of IL-1.beta. (or other cytokines) and the activity of
potential inhibitors. The complexity of this assay system, with its
full complement of lymphoid and inflammatory cell types, spectrum
of plasma proteins and red blood cells is an ideal in vitro
representation of human in vivo physiologic conditions.
Materials:
Pyrogen-free Syringes (.about.30 cc)
[0402] Pyrogen-free sterile vacuum tubes containing lyophilized
Na.sub.2EDTA (4.5 mg/10 ml tube) Human whole blood sample
(.about.30-50 cc) 1.5 ml Eppendorf tubes Test compound stock
solutions (.about.25 mM in DMSO or other solvent) Endotoxin--free
sodium chloride solution (0.9%) and HBSS Lipopolysaccharide (Sigma;
Cat.# L-3012) stock solution at 1 mg/ml in HBSS
IL-1.beta. ELISA Kit (R & D Systems; Cat # DLB50)
TNF.alpha. ELISA Kit (R & D Systems; Cat # DTA50)
[0403] Water bath or incubator
Whole Blood Assay Experimental Procedure:
[0404] Set incubator or water bath at 30.degree. C. Aliquot 0.25 ml
of blood into 1.5 ml eppendorf tubes. Note: be sure to invert the
whole blood sample tubes after every two aliquots. Differences in
replicates may result if the cells sediment and are not uniformly
suspended. Use of a positive displacement pipette will also
minimize differences between replicate aliquots.
[0405] Prepare drug dilutions in sterile pyrogen-free saline by
serial dilution. A dilution series which brackets the apparent
K.sub.i for a test compound determined in an ICE inhibition assay
is generally used for the primary compound screen. For extremely
hydrophobic compounds, prepare compound dilutions in fresh plasma
obtained from the same blood donor or in PBS-containing 5% DMSO to
enhance solubility.
[0406] Add 25 .mu.l test compound dilution or vehicle control and
gently mix the sample. Then add 5.0 .mu.l LPS solution (250 ng/ml
stocked prepared fresh: 5.0 ng/ml final concentration LPS), and mix
again. Incubate the tubes at 30.degree. C. in a water bath for
16-18 hr with occasional mixing. Alternatively, the tubes can be
placed in a rotator set at 4 rpm for the same incubation period.
This assay should be set up in duplicate or triplicate with the
following controls: negative control--no LPS; positive control--no
test inhibitor; vehicle control--the highest concentration of DMSO
or compound solvent used in the experiment. Additional saline is
added to all control tubes to normalize volumes for both control
and experimental whole blood test samples.
[0407] After the incubation period, whole blood samples are
centrifuged for 10 minutes at .about.2000 rpm in the microfuge,
plasma is transferred to a fresh microfuge tube and centrifuged at
1000.times.g to pellet residual platelets if necessary. Plasma
samples may be stored frozen at -70.degree. C. prior to assay for
cytokine levels by ELISA.
ELISA:
[0408] R & D Systems (614 McKinley Place N.E. Minneapolis,
Minn. 55413) Quantikine kits may be used for measurement of
IL-1.beta. and TNF-.alpha.. The assays are performed according to
the manufacturer's directions. IL-1.beta. levels of .about.1-5
ng/ml in positive controls among a range of individuals may be
observed. A 1:200 dilution of plasma for all samples is usually
sufficient for experiments for ELISA results to fall on the linear
range of the ELISA standard curves. It may be necessary to optimize
standard dilutions if you observe differences in the whole blood
assay. Nerad, J. L. et al., J. Leukocyte Biol., 52, pp. 687-692
(1992).
[0409] Selected compounds of this invention have been tested for
inhibition of IL-1.beta. release from whole blood with IC50 values
between 1 .mu.M and 40 .mu.M.
In Vivo Assays
[0410] Compounds of this invention may be tested in in vivo assays
such as those described in WO 99/47545.
[0411] WO 99/47545 and all the other documents cited herein are
hereby incorporated by reference.
[0412] 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.
* * * * *