U.S. patent application number 10/347641 was filed with the patent office on 2004-01-15 for peptide analogs as irreversible interleukin-1beta protease inhibitors.
Invention is credited to Ator, Mark A., Awad, Mohamed M.A., Chaturvedula, Prasad V., Dolle, Roland E., Hoyer, Denton W., Lodge, Eric P., Osifo, Irennegbe K., Prouty, Catherine P., Rinker, James M., Ross, Tina Morgan, Salvino, Joseph M., Schmidt, Stanley J., Singh, Jasbir.
Application Number | 20040009923 10/347641 |
Document ID | / |
Family ID | 26733782 |
Filed Date | 2004-01-15 |
United States Patent
Application |
20040009923 |
Kind Code |
A1 |
Dolle, Roland E. ; et
al. |
January 15, 2004 |
Peptide analogs as irreversible interleukin-1beta protease
inhibitors
Abstract
Disclosed are compounds, compositions and methods for inhibiting
interleukin-1.beta. protease activity, the compounds having the
formula (I).
Inventors: |
Dolle, Roland E.; (King of
Prussia, PA) ; Osifo, Irennegbe K.; (West Chester,
PA) ; Schmidt, Stanley J.; (Chester Springs, PA)
; Hoyer, Denton W.; (Exton, PA) ; Ross, Tina
Morgan; (Audubon, PA) ; Chaturvedula, Prasad V.;
(Exton, PA) ; Prouty, Catherine P.; (Wayne,
PA) ; Awad, Mohamed M.A.; (Frazer, PA) ;
Salvino, Joseph M.; (Schwenksville, PA) ; Rinker,
James M.; (Schwenksville, PA) ; Lodge, Eric P.;
(Pottstown, PA) ; Singh, Jasbir; (Gilbertsville,
PA) ; Ator, Mark A.; (Paoli, PA) |
Correspondence
Address: |
FISH & NEAVE
1251 AVENUE OF THE AMERICAS
50TH FLOOR
NEW YORK
NY
10020-1105
US
|
Family ID: |
26733782 |
Appl. No.: |
10/347641 |
Filed: |
January 16, 2003 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10347641 |
Jan 16, 2003 |
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09421954 |
Oct 20, 1999 |
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6576614 |
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09421954 |
Oct 20, 1999 |
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08679350 |
Jul 10, 1996 |
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5985838 |
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08679350 |
Jul 10, 1996 |
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08371723 |
Jan 12, 1995 |
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08371723 |
Jan 12, 1995 |
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08055051 |
Apr 29, 1993 |
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Current U.S.
Class: |
514/20.1 ;
530/330; 530/331 |
Current CPC
Class: |
C07K 5/06191 20130101;
C07D 333/38 20130101; C07C 235/12 20130101; C07C 311/16 20130101;
C07C 233/51 20130101; C07C 311/19 20130101; A61P 31/00 20180101;
C07C 237/36 20130101; C07D 233/64 20130101; C07D 307/68 20130101;
C07D 295/30 20130101; C07C 271/22 20130101; C07D 295/088 20130101;
C07D 333/36 20130101 |
Class at
Publication: |
514/17 ; 514/18;
514/19; 530/330; 530/331 |
International
Class: |
A61K 038/08; A61K
038/06; A61K 038/04; C07K 005/06; C07K 005/04 |
Claims
What is claimed is:
1. A compound of the formula (I) 17wherein n=0-4; Y= 18m=0,1;
R.sub.3=a singularly or multiply substituted aryl wherein aryl is a
phenyl or naphthyl ring wherein the substituents are independently
selected from the group consisting of: (1) H (2) halogen (3) OH (4)
CF.sub.3 (5) NO.sub.2 (6) OR.sub.5 (7) COR.sub.9 (8)
NR.sub.6COR.sub.10 (9) CONR.sub.5R.sub.6 (10)
SO.sub.2NR.sub.5R.sub.6 (11) SO.sub.2R.sub.6 (12) COOR.sub.11
19(14) lower alkyl and lower cycloalkyl R.sub.5=(1) lower straight
chain or branched alkyl, lower cycloalkyl (2)
(CR.sub.6R.sub.7).sub.0-6-aryl (3)
(CR.sub.6R.sub.7).sub.0-6-heteroaryl or (4)
(CR.sub.6R.sub.7).sub.2-6-R.sub.8; R.sub.6 and R.sub.7 are
independently H, lower straight chain or branched alkyl, benzyl,
aryl, cycloalkyl and aryl is defined as above and heteroaryl
includes pyridyl, thienyl, furyl, thiazolyl, imidazolyl, pyrazolyl,
isoxazolyl, benzimidazolyl, pyrazinyl, pyrimiidyl, quinolyl,
isoquinolyl, isothiazolyl, benzofuranyl, isoxazolyl, triazinyl and
tetrazolyl; R.sub.8=(1) OCH.sub.2CH.sub.2OR.sub.6 (2)
OCH.sub.2CH.sub.2NR.sub.6R.sub.- 7 (3)
NR.sub.6CH.sub.2CO.sub.2R.sub.6 20(6) NR.sub.6R.sub.7 wherein
R.sub.6 and R.sub.7 are as above defined; R.sub.9=(1) lower
straight chain or branched alkyl, lower cycloalkyl (2)
(CR.sub.6R.sub.7).sub.0-6-a- ryl; (3)
(CR.sub.6R.sub.7).sub.0-6-heteroaryl; or (4)
(CR.sub.6R.sub.7).sub.0-6-R.sub.8, wherein R.sub.6, R.sub.7 and
R.sub.8 are as above defined; R.sub.10=(1) R.sub.9 (2) OR.sub.11
(3) NR.sub.6R.sub.11, wherein R.sub.11=(1) lower straight chain or
branched alkyl, lower cycloalkyl (2)
(CR.sub.6R.sub.7).sub.1-6-aryl; (3)
(CR.sub.6R.sub.7).sub.1-6-heteroaryl; or (4)
(CR.sub.6R.sub.7).sub.2-6-R.- sub.8, and R.sub.6, R.sub.7 and
R.sub.8 are as above defined; R.sub.4=H or deuterium; R.sub.2=(1)
OR.sub.6 (2) NR.sub.6OR.sub.7 or (3) NR.sub.6R.sub.7, and R.sub.6
and R.sub.7 are as above-defined; A=(1) an amino acid of the
formula (II) 21wherein R.sub.6 and R.sub.7 are as defined above;
R.sub.12 is independently (1) H or (2)
(CR.sub.6R.sub.7).sub.1-6-R.sub.13, and R.sub.6 and R.sub.7 are as
above-defined; R.sub.13=(1) H (2) F (3) CF.sub.3 (4) OH (5)
OR.sub.11 (6) NR.sub.6R.sub.14 (7) cycloalkyl (8) aryl (9)
heteroaryl (10) SH (11) SR.sub.11 (12) CONR.sub.5R.sub.6 (13)
COOR.sub.5 or 22R.sub.14=(1) R.sub.7 (2) COR.sub.10 (3)
SO.sub.2NR.sub.5R.sub.6 or 23or A=(2) an amino acid selected from
the group consisting of 2425R.sub.1 is an acyl group of the formula
(III) 26wherein R.sub.12 is (1) OR.sub.5 (2) NR.sub.5R.sub.6 (3)
R.sub.5 (4) --CH.dbd.CHR.sub.5 27wherein R.sub.15 single bond,
(CH.sub.2).sub.2-6--NR.sub.6--, (CH.sub.2).sub.2-6--O-- and R.sub.5
and R.sub.6 are as above defined; or a sulfonyl group of the
formula (IV) 28wherein R.sub.16 is (1) R.sub.5 29wherein R.sub.5
and R.sub.6 are as above-defined.
2. A compound according to claim 1 selected from the group
consisting of: N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone, N-Benzyloxycarbonyl-L-aspartic
acid 2,6-difluorophenoxymethyl ketone,
N-Benzyloxycarbonyl-L-aspartic acid
2,6-ditrifluoromethylbenzoyloxymethyl ketone,
N-Benzyloxycarbonyl-L-aspartic acid 2,6-dichlorophenoxymethyl
ketone and N-Benzyloxycarbonyl-L-aspartic acid
2-fluoro-4-(N-morpholinyls- ulfonamido)phenoxymethyl ketone.
3. A compound according to claim 1 selected from the group
consisting of: N-Benzyloxycarbonyl-L-aspartic acid
2-chloro-4-(N-thiomorpholinyl sulfonamido)phenoxymethyl ketone,
N-Benzyl oxycarbonyl-L-aspartic acid
2,6-dichloro-3-(2-N-morpholinylethoxy) benzyloxymethyl ketone,
N-Benzyloxycarbonyl-L-aspartic acid 2,6-dimethoxybenzoyloxy methyl
ketone, N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichloro-3-(benzyloxy)ben- zyloxymethyl ketone and
N-Benzyloxycarbonyl-L-aspartic acid
2-acetamido-6-chlorobenzoyloxymethyl ketone.
4. A compound according to claim 1 selected from the group
consisting of: N-Benzyloxycarbonyl-L-aspartic acid
2,6-difluorobenzoyloxymethyl ketone, N-Benzyloxycarbonyl-L-aspartic
acid 3-(N-butylsulfonamido)-2,6-dichlorobe- nzoyloxymethyl ketone,
N-Benzyloxycarbonyl-L-aspartic acid 2,6-dichloro-3-sulfonamido
benzoylmethyl ketone, N-Benzyloxycarbonyl-L-as- partic acid
3-(N-benzylsulfonamido)-2,6-dichlorobenzoyloxymethyl ketone and
N-Benzyloxycarbonyl-L-aspartic acid
3-(N-[2-aminoacetamidoyl]sulfonam-
ido)-2,6-dichlorobenzoyloxymethyl ketone.
5. A compound according to claim 1 selected from the group
consisting of: N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichloro-3-(N-morpholinylsulfonam- ido)benzoyloxymethyl ketone,
N-Methoxycarbonyl-L-alanine-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone, N-(2-thienyl)
carbonyl-L-aspartic acid 2,6-dichlorobenzoyloxy-methyl ketone,
N-Methoxycarbonyl-glycine-L-as- partic acid
2,6-dichlorobenzoyloxymethyl ketone and
N-Methoxycarbonyl-L-phenylalanine-L-aspartic acid
2,6-dichlorobenzoyloxym- ethyl ketone.
6. A compound according to claim 1 selected from the group
consisting of:
N-Methoxycarbonyl-L-.beta.-(2-thienyl)alanine-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone,
N-Methoxycarbonyl-L-valine-L-asparti- c acid
2,6-dichlorobenzoyloxymethyl ketone,
N-Methoxycarbonyl-L-histidine-- L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone,
N-Benzyloxycarbonyl-L-valine-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone and
N-Benzyloxycarbonyl-L-alanine-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone.
7. A compound according to claim 1 selected from the group
consisting of: N-Benzyloxycarbonyl-L-valine-L-alanine-L-aspartic
acid 2,6-dichlorobenzoyloxymethyl ketone,
N-(2-furonyl)carbonyl-L-aspartic acid 2,6-dichlorobenzoyloxymethyl
ketone, N-(2-furonyl)carbonyl-L-asparti- c acid
2,6-dichloro-3-(N-morpholinylsulfonamido)-benzoyloxymethyl ketone,
N-(3-phenylpropionyl)-L-aspartic acid 2,6-dichlorobenzoyloxymethyl
ketone, N-Methoxycarbonyl-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone,
N-(4-(N,N-dimethylaminomethyl)benzoyl-L-aspartic acid
2,6-dichloro-benzyloxymethyl ketone, N-benzyloxycarbonyl-D-aspartic
acid 2,6-dichlorobenzoyloxymethyl ketone,
N-(2-[2,6-dichlorobenzoyloxy])acetyl- -L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone and
N-Benzyloxycarbonyl-L-valine-L-aspartic acid
N-Benzyloxycarbonyl-L-valine- -L-aspartic acid
4-(N,N-diethyl-sulfonamido)-2,3,5,6-tetrafluoro-phenoxyme- thyl
ketone.
8. A pharmaceutical composition for inhibiting interleukin 1.beta.
protease comprising a compound of the formula (I) 30wherein n=0-4;
Y= 31m=0,1; R.sub.3=a singularly or multiply substituted aryl
wherein aryl is a phenyl or naphthyl ring wherein the substituents
are independently selected from the group consisting of: (1) H (2)
halogen (3) OH (4) CF.sub.3 (5) NO.sub.2 (6) OR.sub.5 (7) COR.sub.9
(8) NR.sub.6COR.sub.10 (9) CONR.sub.5R.sub.6 (10)
SO.sub.2NR.sub.5R.sub.6 (11) SO.sub.2R.sub.6 (12) COOR.sub.11
32(14) lower alkyl and lower cycloalkyl R.sub.5=(1) lower straight
chain or branched alkyl, lower cycloalkyl (2)
(CR.sub.6R.sub.7).sub.0-6-aryl (3)
(CR.sub.6R.sub.7).sub.0-6-heteroaryl or (4)
(CR.sub.6R.sub.7).sub.2-6-R.sub.8; R.sub.6 and R.sub.7 are
independently H, lower straight chain or branched alkyl, benzyl,
aryl, cycloalkyl and aryl is defined as above and heteroaryl
includes pyridyl, thienyl, furyl, thiazolyl, imidazolyl, pyrazolyl,
isoxazolyl, benzimidazolyl, pyrazinyl, pyrimidyl, quinolyl,
isoquinolyl, isothiazolyl, benzofuranyl, isoxazolyl, triazinyl and
tetrazolyl; R.sub.8=(1) OCH.sub.2CH.sub.2OR.sub.6 (2)
OCH.sub.2CH.sub.2NR.sub.6R.sub.- 7 (3)
NR.sub.6CH.sub.2CO.sub.2R.sub.6 33(6) NR.sub.6R.sub.7 wherein
R.sub.6 and R.sub.7 are as above defined; R.sub.9=(1) lower
straight chain or branched alkyl, lower cycloalkyl (2)
(CR.sub.6R.sub.7).sub.0-6-a- ryl; (3)
(CR.sub.6R.sub.7).sub.0-6-heteroaryl; or (4)
(CR.sub.6R.sub.7).sub.0-6-R.sub.8, wherein R.sub.6, R.sub.7 and
R.sub.8 are as above defined; R.sub.10=(1) R.sub.9 (2) OR.sub.11
(3) NR.sub.6R.sub.11, wherein R.sub.11=(1) lower straight chain or
branched alkyl, lower cycloalkyl (2)
(CR.sub.6R.sub.7).sub.1-6-aryl; (3)
(CR.sub.6R.sub.7).sub.1-6-heteroaryl; or (4)
(CR.sub.6R.sub.7).sub.2-6-R.- sub.8, and R.sub.6, R.sub.7 and
R.sub.8 are as above defined; R.sub.4=H or deuterium; R.sub.2=(1)
OR.sub.6 (2) NR.sub.6OR.sub.7 or (3) NR.sub.6R.sub.7, and R.sub.6
and R.sub.7 are as above-defined; A=(1) an amino acid of the
formula (II) 34wherein R.sub.6 and R.sub.7 are as defined above;
R.sub.12 is independently (1) H or (2)
(CR.sub.6R.sub.7).sub.1-6-R.sub.13, and R.sub.6 and R.sub.7 are as
above-defined; R.sub.13=(1) H (2) F (3) CF.sub.3 (4) OH (5)
OR.sub.11 (6) NR.sub.6R.sub.14 (7) cycloalkyl (8) aryl (9)
heteroaryl (10) SH (11) SR.sub.11 (12) CONR.sub.5R.sub.6 (13)
COOR.sub.5 or 35R.sub.14=(1) R.sub.7 (2) COR.sub.10 (3)
SO.sub.2NR.sub.5R.sub.6 or 36or A=(2) an amino acid selected from
the group consisting of 3738R.sub.1 is an acyl group of the formula
(III) 39wherein R.sub.12 is (1) OR.sub.5 (2) NR.sub.5R.sub.6 (3)
R.sub.5 (4) --CH.dbd.CHR.sub.5 40wherein R.sub.15=single bond,
(CH.sub.2).sub.2-6--NR.sub.6--, (CH.sub.2).sub.2-6--O-- and R.sub.5
and R.sub.6 are as above defined; or a sulfonyl group of the
formula (IV) 41wherein R.sub.16 is (1) R.sub.5 42wherein R.sub.5
and R.sub.6 are as above-defined.
9. The pharmaceutical composition of claim 8 wherein said compound
is selected from the group consisting of:
N-Benzyloxycarbonyl-L-aspartic acid 2,6-dichlorobenzoyloxy-methyl
ketone, N-Benzyloxycarbonyl-L-aspartic acid
2,6-difluorophenoxymethyl ketone, N-Benzyloxycarbonyl-L-aspartic
acid 2,6-ditrifluoro methyl benzoyloxymethyl ketone,
N-Benzyloxycarbonyl-L-aspartic acid 2,6-dichlorophenoxymethyl
ketone and N-Benzyloxycarbonyl-L-aspartic acid
2-fluoro-4-(N-morpholinylsulfonamido)- phenoxymethyl ketone.
10. The pharmaceutical composition of claim 8 wherein said compound
is selected from the group consisting of:
N-Benzyloxycarbonyl-L-aspartic acid
2-chloro-4-(N-thiomorpholinyl-sulfonamido )phenoxymethyl ketone,
N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichloro-3-(2-N-morpholinylethoxy- )benzyloxymethyl ketone,
N-Benzyloxycarbonyl-L-aspartic acid 2,6-dimethoxybenzoyloxy methyl
ketone, N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichloro-3-(benzyloxy)-benzyloxymethyl ketone and
N-Benzyloxycarbonyl-L-aspartic acid
2-acetamido-6-chlorobenzoyloxymethyl ketone.
11. The pharmaceutical composition of claim 8 wherein said compound
is selected from the group consisting of:
N-Benzyloxycarbonyl-L-aspartic acid 2,6-difluorobenzoyloxymethyl
ketone, N-Benzyloxycarbonyl-L-aspartic acid
3-(N-butylsulfonamido)-2,6-dichlorobenzoyloxymethyl ketone,
N-Benzyloxycarbonyl-L-aspartic acid 2,6-dichloro-3-sulfonamido
benzoylmethyl ketone, N-Benzyloxycarbonyl-L-aspartic acid
3-(N-benzylsulfonamido)-2,6-dichlorobenzoyloxymethyl ketone and
N-Benzyloxycarbonyl-L-aspartic acid
3-(N-[2-aminoacetamidoyl]sulfonamido)-
-2,6-dichlorobenzoyloxymethyl ketone.
12. The pharmaceutical composition of claim 8 wherein said compound
is selected from the group consisting of:
N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichloro-3-(N-morpholinylsulfonamido) benzoyloxymethyl ketone,
N-Methoxycarbonyl-L-alanine-L-aspartic acid
2,6-dichlorobenzoyloxy-methyl ketone, N-(2-thienyl)
carbonyl-L-aspartic acid 2,6-dichlorobenzoyloxy-met- hyl ketone,
N-Methoxycarbonyl-glycine-L-aspartic acid
2,6-dichlorobenzoyloxy-methyl ketone and
N-Methoxycarbonyl-L-phenylalanin- e-L-aspartic acid
2,6-dichloro-benzoyloxymethyl ketone.
13. The pharmaceutical composition of claim 8 wherein said compound
is selected from the group consisting of:
N-Methoxycarbonyl-L-.beta.-(2-thie- nyl)alanine-L-aspartic acid
2,6-di-chlorobenzoyloxymethyl ketone,
N-Methoxycarbonyl-L-valine-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone,
N-Methoxycarbonyl-L-histidine-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone,
N-Benzyloxycarbonyl-L-valine-L-aspar- tic acid
2,6-dichlorobenzoyloxymethyl ketone and N-Benzyloxycarbonyl-L-ala-
nine-L-aspartic acid 2,6-dichlorobenzoyloxymethyl ketone.
14. The pharmaceutical composition of claim 8 wherein said compound
is selected from the group consisting of:
N-Benzyloxycarbonyl-L-valine-L-ala- nine-L-aspartic acid
2,6-di-chlorobenzoyloxymethyl ketone,
N-(2-furonyl)carbonyl-L-aspartic acid 2,6-dichlorobenzoyloxymethyl
ketone, N-(2-furonyl)carbonyl-L-aspartic acid
2,6-dichloro-3-(N-morpholin- ylsulfonamido)-benzoyloxymethyl
ketone, N-(3-phenylpropionyl)-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone, N-Methoxycarbonyl-L-aspartic
acid 2,6-dichlorobenzoyloxymethyl ketone,
N-(4-(N,N-dimethylaminomethyl)b- enzoyl-L-aspartic acid
2,6-dichloro-benzoyloxymethyl ketone,
N-benzyloxycarbonyl-D-aspartic acid 2,6-dichlorobenzoyloxymethyl
ketone, N-(2-[2,6-dichlorobenzoyloxy])acetyl-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone and
N-Benzyloxycarbonyl-L-valine-L-as- partic acid
4-(N,N-diethyl-sulfonamido)-2,3,5,6-tetrafluorophenoxymethyl
ketone.
15. A method of inhibiting interleukin-1.beta. protease activity in
a mammal in need of such treatment comprising administering to said
mammal an effective inhibitory amount of a pharmaceutical
composition comprising a compound of the formula (I) or a
pharmaceutically acceptable salt thereof 43wherein n=0-4; Y=
44m=0,1; R.sub.3=a singularly or multiply substituted aryl wherein
aryl is a phenyl or naphthyl ring wherein the substituents are
independently selected from the group consisting of: (1) H (2)
halogen (3) OH (4) CF.sub.3 (5) NO.sub.2 (6) OR.sub.5 (7) COR.sub.9
(8) NR.sub.6COR.sub.10 (9) CONR.sub.5R.sub.6 (10)
SO.sub.2NR.sub.5R.sub.6 (11) SO.sub.2R.sub.6 (12) COOR.sub.11
45(14) lower alkyl and lower cycloalkyl R.sub.5=(1) lower straight
chain or branched alkyl, lower cycloalkyl (2)
(CR.sub.6R.sub.7).sub.0-6-aryl (3) (CR.sub.6R.sub.7).sub.0-
-6-heteroaryl or (4) (CR.sub.6R.sub.7).sub.2-6-R.sub.8; R.sub.6 and
R.sub.7 are independently H, lower straight chain or branched
alkyl, benzyl, aryl, cycloalkyl and aryl is defined as above and
heteroaryl includes pyridyl, thienyl, furyl, thiazolyl, imidazolyl,
pyrazolyl, isoxazolyl, benzimidazolyl, pyrazinyl, pyrimidyl,
quinolyl, isoquinolyl, isothiazolyl, benzofuranyl, isoxazolyl,
triazinyl and tetrazolyl; R.sub.8=(1) OCH.sub.2CH.sub.2OR.sub.6 (2)
OCH.sub.2CH.sub.2NR.sub.6R.sub.- 7 (3)
NR.sub.6CH.sub.2CO.sub.2R.sub.6 46(6) NR.sub.6R.sub.7 wherein
R.sub.6 and R.sub.7 are as above defined; R.sub.9=(1) lower
straight chain or branched alkyl, lower cycloalkyl (2)
(CR.sub.6R.sub.7).sub.0-6-a- ryl; (3)
(CR.sub.6R.sub.7).sub.0-6-heteroaryl; or (4)
(CR.sub.6R.sub.7).sub.0-6-R.sub.8, wherein R.sub.6, R.sub.7 and
R.sub.8 are as above defined; R.sub.10=(1) R.sub.9 (2) OR.sub.11
(3) NR.sub.6R.sub.11, wherein R.sub.11=(1) lower straight chain or
branched alkyl, lower cycloalkyl (2)
(CR.sub.6R.sub.7).sub.1-6-aryl; (3)
(CR.sub.6R.sub.7).sub.1-6-heteroaryl; or (4)
(CR.sub.6R.sub.7).sub.2-6-R.- sub.8, and R.sub.6, R.sub.7 and
R.sub.8 are as above defined; R.sub.4=H or deuterium; R.sub.2=(1)
OR.sub.6 (2) NR.sub.6OR.sub.7 or (3) NR.sub.6R.sub.7, and R.sub.6
and R.sub.7 are as above-defined; A=(1) an amino acid of the
formula (II) 47wherein R.sub.6 and R.sub.7 are as defined above;
R.sub.12 is independently (1) H or (2)
(CR.sub.6R.sub.7).sub.1-6-R.sub.13, and R.sub.6 and R.sub.7 are as
above-defined; R.sub.13=(1) H (2) F (3) CF.sub.3 (4) OH (5)
OR.sub.11 (6) NR.sub.6R.sub.14 (7) cycloalkyl (8) aryl (9)
heteroaryl (10) SH (11) SR.sub.11 (12) CONR.sub.5R.sub.6 (13)
COOR.sub.5 or 48R.sub.14=(1) R.sub.7 (2) COR.sub.10 (3)
SO.sub.2NR.sub.5R.sub.6 or 49or A=(2) an amino acid selected from
the group consisting of 50R.sub.1 is an acyl group of the formula
(III) 51wherein R.sub.12 is (1) OR.sub.5 (2) NR.sub.5R.sub.6 (3)
R.sub.5 (4) --CH.dbd.CHR.sub.5 52wherein R.sub.15=single bond,
(CH.sub.2).sub.2-6--NR.sub.6--, (CH.sub.2).sub.2-6--O-- and R.sub.5
and R.sub.6 are as above defined; or a sulfonyl group of the
formula (IV) 53wherein R.sub.16 is (1) R.sub.5 54wherein R.sub.5
and R.sub.6 are as above-defined.
16. The method of claim 15 wherein said compound is selected from
the group consisting of: N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichlorobenzoyloxy-methyl ketone,
N-Benzyloxycarbonyl-L-aspartic acid 2,6-difluorophenoxymethyl
ketone, N-Benzyloxycarbonyl-L-aspartic acid 2,6-ditrifluoromethyl
benzoyloxymethyl ketone, N-Benzyloxycarbonyl-L-aspa- rtic acid
2,6-dichlorophenoxymethyl ketone and N-Benzyloxycarbonyl-L-aspar-
tic acid 2-fluoro-4-(N-morpholinylsulfonamido)phenoxymethyl
ketone.
17. The method of claim 15 wherein said compound is selected from
the group consisting of: N-Benzyloxycarbonyl-L-aspartic acid
2-chloro-4-(N-thiomorpholinyl-sulfonamido)phenoxymethyl ketone,
N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichloro-3-(2-N-morpholinylethoxy- )benzyloxymethyl ketone,
N-Benzyloxycarbonyl-L-aspartic acid 2,6-dimethoxybenzoyloxy methyl
ketone, N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichloro-3-(benzyloxy)benzyloxymethyl ketone and
N-Benzyloxycarbonyl-L-aspartic acid
2-acetamido-6-chlorobenzoyloxymethyl ketone.
18. The method of claim 15 wherein said compound is selected from
the group consisting of: N-Benzyloxycarbonyl-L-aspartic acid
2,6-difluorobenzoyloxymethyl ketone, N-Benzyloxycarbonyl-L-aspartic
acid 3-(N-butylsulfonamido)-2,6-dichlorobenzoyloxymethyl ketone,
N-Benzyloxycarbonyl-L-aspartic acid 2,6-dichloro-3-sulfonamido
benzoylmethyl ketone, N-Benzyloxycarbonyl-L-aspartic acid
3-(N-benzylsulfonamido)-2,6-dichlorobenzoyloxymethyl ketone and
N-Benzyloxycarbonyl-L-aspartic acid
3-(N-[2-aminoacetamidoyl]sulfonamido)-
-2,6-dichlorobenzoyloxymethyl ketone.
19. The method of claim 15 wherein said compound is selected from
the group consisting of: N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichloro-3-(N-morpholinylsulfonamido) benzoyloxymethyl ketone,
N-Methoxycarbonyl-L-alanine-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone, N-(2-thienyl)
carbonyl-L-aspartic acid 2,6-dichlorobenzoyloxy-met- hyl ketone,
N-Methoxycarbonyl-glycine-L-aspartic acid
2,6-dichlorobenzoyloxy-methyl ketone and
N-Methoxycarbonyl-L-phenylalanin- e-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone.
20. The method of claim 15 wherein said compound is selected from
the group consisting of:
N-Methoxycarbonyl-L-.beta.-(2-thienyl)alanine-L-aspa- rtic acid
2,6-di-chlorobenzoyloxymethyl ketone, N-Methoxycarbonyl-L-valine-
-L-aspartic acid 2,6-dichlorobenzoyloxymethyl ketone,
N-Methoxycarbonyl-L-histidine-L-aspartic acid
2,6-dichlorobenzoyloxymethy- l ketone,
N-Benzyloxycarbonyl-L-valine-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone and
N-Benzyloxycarbonyl-L-alanine-L-a- spartic acid
2,6-dichlorobenzoyloxymethyl ketone.
21. The method of claim 15 wherein said compound is selected from
the group consisting of:
N-Benzyloxycarbonyl-L-valine-L-alanine-L-aspartic acid
2,6-di-chlorobenzoyloxymethyl ketone,
N-(2-furonyl)carbonyl-L-aspart- ic acid
2,6-dichlorobenzoyloxymethyl ketone, N-(2-furonyl)carbonyl-L-aspar-
tic acid 2,6-dichloro-3-(N-morpholinylsulfonamido)benzoyloxymethyl
ketone, N-(3-phenylpropionyl)-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone, N-Methoxycarbonyl-L-aspartic
acid 2,6-dichlorobenzoyloxymethyl ketone,
N-(4-(N,N-dimethylaminomethyl)benzoyl-L-aspartic acid
2,6-dichloro-benzoyloxymethyl ketone,
N-benzyloxycarbonyl-D-aspartic acid 2,6-dichlorobenzoyloxymethyl
ketone, N-(2-[2,6-dichlorobenzoyloxy])acetyl- -L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone and
N-Benzyloxycarbonyl-L-valine-L-aspartic acid
4-(N,N-diethyl-sulfonamido)-- 2,3,5,6-tetrafluorophenoxymethyl
ketone.
Description
BACKGROUND OF THE INVENTION
[0001] 1. Field of the Invention
[0002] This invention relates to peptide analogs that are
interleukin-1.beta. protease inhibitors. More particularly, the
invention provides .alpha.-substituted methyl ketones derived from
aspartic acid and the closed hemi-ketal forms thereof as inhibitors
of interleukin 1-.beta. protease.
[0003] 2. Reported Developments
[0004] Enzymes involved in the catalytic degradation of proteins by
hydrolyzing peptide bonds are known as proteases or proteinases.
Proteinases are believed to be involved in various disease states
including inflammation, metastasis, tissue damage, bone resorption
and muscle degeneration in dystrophic diseases. Proteinases are
divided into classes according to their catalytic mechanisms, such
as serine-, cystein-, aspartic- and metallo-proteinases. For each
class of proteinases, the catalytic site of the enzyme lies in the
cleft on the surface of the enzymes in which reside the specificity
subsites that bind amino acid side chains and the polypeptide
backbone. In designing proteinase inhibitors, it is important to
optimize the subsite binding characteristics with appropriate amino
acid substrate analogs.
[0005] This invention relates to peptide substrates modified with
affinity labels that inhibit interleukin-1.beta. protease
(hereinafter IL-1.beta. protease).
[0006] These inhibitors are thought to act by alkylating the
cysteine sulfhydryl group (cys 285) within the catalytic site of
IL-1.beta. protease. Affinity labeling has been used since the
1960's to prepare irreversible peptide-based inhibitors which act
to alkylate the active sites of cysteine proteases. A variety of
affinity labels and amino acid sequences have been synthesized to
improve the binding of these modified peptide inhibitors to the
enzyme's active site. These affinity labels include peptidyl
halomethyl ketones, peptidyl diazomethyl ketones, epoxysuccinyl
peptides and peptidyl methylsulphonium salts as reviewed by D. Rich
in Chapter 4 of "Proteinase Inhibitors", Barret, A. J. and
Salvesen, G., eds., Elsevier, 1986. More recently, peptide
acyloxymethyl and aryloxymethyl ketons have also been described as
affinity lables (Krantz, A. et al, Biochemisty, 30, p. 4678-4687,
1991). Current research (see for example European Patent
Application, Pub. No. 015,748 A2; PCT International Publication No.
WO 91/15577; Chapman, K. T., Biorganic & Medicinal Chem. Lett.
1992, 2, 613-618) has been directed towards understanding the
enzyme binding specificity requirements in designing novel small
molecular weight protease inhibitors that are efficacious, safe and
have specificity for IL-1.beta. protease which is believed to play
an important role in many disease states (see Epstein, F. H., New
Engl. Jrl. of Med., 328, p. 106-113, 1993).
[0007] Disease states in which IL-1.beta. protease inhibitors may
be useful as therapeutic agents include: infectious diseases, such
as meningitis and salpingitis; septic shock, respiratory diseases;
inflammatory conditions, such as arthritis, cholangitis, colitis,
encephalitis, endocerolitis, hepatitis, pancreatitis and
reperfusion injury, immune-based diseases, such as
hypersensitivity; auto-immune diseases, such as multiple sclerosis;
bone diseases; and certain tumors.
[0008] It is an object of the present invention to provide novel
peptidyl substrate analogs modified with electronegative leaving
groups that bind at the active site of IL-1.beta. protease and
inhibit IL-1.beta. protease activity. IL-1.beta. protease cleaves a
biologically inactive 34 kD precursor of IL-1.beta. to form the
biologically active 17 kD cytokine. This cleavage occurs at the
peptidyl sequence of Val-His-Asp/-Ala-Pro-Val- .
[0009] It is another object of the present invention to provide
compositions comprising the above-referred to compounds.
[0010] It is a further object of the present invention to provide a
method of use of the composition for the treatment of the
above-identified disease states.
SUMMARY OF THE INVENTION
[0011] According to the present invention, there is provided a
compound of the formula (I) and a pharmaceutically acceptable salt
thereof: 1
[0012] wherein
[0013] n=0-4;
[0014] Y= 2
[0015] m=0,1;
[0016] R.sub.3=a singularly or multiply substituted aryl wherein
aryl is a phenyl or naphthyl ring wherein the substituents are
independently selected from the group consisting of:
[0017] (1) H
[0018] (2) halogen
[0019] (3) OH
[0020] (4) CF.sub.3
[0021] (5) NO.sub.2
[0022] (6) OR.sub.5
[0023] (7) COR.sub.9
[0024] (8) NR.sub.6COR.sub.10
[0025] (9) CONR.sub.5R.sub.6
[0026] (10) SO.sub.2NR.sub.5R.sub.6
[0027] (11) SO.sub.2R.sub.6
[0028] (12) COOR.sub.11 3
[0029] (14) lower alkyl and lower cycloalkyl
[0030] R.sub.5=
[0031] (1) lower straight chain or branched alkyl, lower
cycloalkyl
[0032] (2) (CR.sub.6R.sub.7).sub.0-6-aryl
[0033] (3) (CR.sub.6R.sub.7).sub.0-6-heteroaryl or
[0034] (4) (CR.sub.6R.sub.7).sub.2-6-R.sub.8;
[0035] R.sub.6 and R.sub.7 are independently H, lower straight
chain or branched alkyl, benzyl, aryl, cycloalkyl and aryl is
defined as above and heteroaryl includes pyridyl, thienyl, furyl,
thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, benzimidazolyl,
pyrazinyl, pyrimidyl, quinolyl, isoquinolyl, isothiazolyl,
benzofuranyl, isoxazolyl, triazinyl and tetrazolyl;
[0036] R.sub.8=
[0037] (1) OCH.sub.2CH.sub.2OR.sub.6
[0038] (2) OCH.sub.2CH.sub.2NR.sub.6R.sub.7
[0039] (3) NR.sub.6CH.sub.2CO.sub.2R.sub.6 4
[0040] (6) NR.sub.6R.sub.7 wherein R.sub.6 and R.sub.7 are as above
defined;
[0041] R.sub.9=
[0042] (1) lower straight chain or branched alkyl, lower
cycloalkyl
[0043] (2) (CR.sub.6R.sub.7).sub.0-6-aryl;
[0044] (3) (CR.sub.6R.sub.7).sub.0-6-heteroaryl; or
[0045] (4) (CR.sub.6R.sub.7).sub.0-6-R.sub.8, wherein R.sub.6,
R.sub.7 and R.sub.8 are as above defined;
[0046] R.sub.10=
[0047] (1) R.sub.9
[0048] (2) OR.sub.11
[0049] (3) NR.sub.6R.sub.11,
[0050] wherein
[0051] R.sub.11=
[0052] (1) lower straight chain or branched alkyl, lower
cycloalkyl
[0053] (2) (CR.sub.6R.sub.7).sub.1-6-aryl;
[0054] (3) (CR.sub.6R.sub.7).sub.1-6-heteroaryl; or
[0055] (4) (CR.sub.6R.sub.7).sub.2-6-R.sub.8, and R.sub.6, R.sub.7
and R.sub.8 are as above defined;
[0056] R.sub.4=
[0057] H or deuterium;
[0058] R.sub.2=
[0059] (1) OR.sub.6
[0060] (2) NR.sub.6OR.sub.7 or
[0061] (3) NR.sub.6R.sub.7, and R.sub.6 and R.sub.7 are as
above-defined;
[0062] A=
[0063] (1) an amino acid of the formula (II) 5
[0064] wherein R.sub.6 and R.sub.7 are as defined above;
[0065] R.sub.12 is independently
[0066] (1) H or
[0067] (2) (CR.sub.6R.sub.7).sub.1-6-R.sub.13, and R.sub.6 and
R.sub.7 are as above-defined;
[0068] R.sub.13=
[0069] (1) H
[0070] (2) F
[0071] (3) CF.sub.3
[0072] (4) OH
[0073] (5) OR.sub.11
[0074] (6) NR.sub.6R.sub.14
[0075] (7) cycloalkyl
[0076] (8) aryl
[0077] (9) heteroaryl
[0078] (10) SH
[0079] (11) SR.sub.11
[0080] (12) CONR.sub.5R.sub.6
[0081] (13) COOR.sub.5 or 6
[0082] R.sub.14=
[0083] (1) R.sub.7
[0084] (2) COR.sub.10
[0085] (3) SO.sub.2NR.sub.5R.sub.6 or 7
[0086] or
[0087] A=
[0088] (2) an amino acid selected from the group consisting of
89
[0089] R.sub.1 is an acyl group of the formula (III) 10
[0090] wherein
[0091] R.sub.12 is
[0092] (1) OR.sub.5
[0093] (2) NR.sub.5R.sub.6
[0094] (3) R.sub.5
[0095] (4) --CH.dbd.CHR.sub.5 11
[0096] wherein R.sub.15 single bond,
(CH.sub.2).sub.2-6--NR.sub.6--, (CH.sub.2).sub.2-6--O-- and
[0097] R.sub.5 and R.sub.6 are as above defined; or
[0098] a sulfonyl group of the formula (IV) 12
[0099] wherein
[0100] R.sub.16 is
[0101] (1) R.sub.5 13
[0102] wherein R.sub.5 and R.sub.6 are as above-defined.
[0103] As used herein the term pharmaceutically acceptable salts
include the acid and base addition salts.
[0104] The term acid addition salts refers to those salts which
retain the biological effectiveness and properties of the free
bases and which are not biologically or otherwise undesirable,
formed with inorganic acids such as hydrochloric acid, hydrobromic
acid, sulfuric acid, nitric acid, phosphoric acid and the like, and
organic acids such as acetic acid, propionic acid, glycolic acid,
pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic
acid, fumaric acid, tartaric acid, citric acid, benzoic acid,
cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic
acid, p-toluenesulfonic acid, salicylic acid and the like.
[0105] The term base addition salts include those derived from
inorganic bases such as sodium, potassium, lithium, ammonium,
calcium, magnesium, iron, zinc, copper, manganese, aluminum salts
and the like. Particularly preferred are the ammonium, potassium,
sodium, calcium and magnesium salts derived from pharmaceutically
acceptable organic non-toxic bases include salts of primary,
secondary, and tertiary amines, substituted amines including
naturally occurring substituted amines, cyclic amines and basic ion
exchange resins, such as isopropylamine, trimethylamine,
diethylamine, triethylamine, tripropylamine, ethanolamine,
2-dimethylaminoethanol, 2-diethylaminoethanol, trimethamine,
dicyclohexylamine, lysine, arginine, histidine, caffeine,
procaines, hydrabamine, choline, betaine, ethylenediamine,
glucosamine, methylglucamine, theobromine, purines, piperazine,
piperidine, N-ethylpiperidine, polyamine resins and the like.
Particularly preferred organic non-toxic bases are isopropylamine,
diethylamine, ethanolamine, trimethamine, dicyclohexylamine,
choline and caffeine.
[0106] "Alkyl" means a saturated or unsaturated aliphatic
hydrocarbon which may be either straight- or branched-chain.
Preferred groups have no more than about 12 carbon atoms and may be
methyl, ethyl and structural isomers of propyl, butyl, pentyl,
hexyl, heptyl, octyl, nonyl, decyl, undecyl and dodecyl.
[0107] "Lower alkyl" means an alkyl group as above, having 1 to 7
carbon atoms. Suitable lower alkyl groups are methyl, ethyl,
n-propyl, isopropyl, butyl, tert-butyl, n-pentyl, neopentyl,
n-hexyl, and n-heptyl.
[0108] "Substituted phenyl" means a phenyl group in which one or
more of the hydrogens has been replaced by the the same or
different substituents including halo, lower alkyl, nitro, amino,
acylamino, hydroxyl, lower alkoxy, aryl, heteroaryl, lower alkoxy,
alkylsulfonyl, trifluoromethyl, morpholinoethoxy,
morpholino-sulfonyl, and carbobenzoxy-methyl sulfamoyl.
[0109] "Halogen" means chloride, fluoride, bromide or iodide.
[0110] "Lower cycloalkyl" means cycloalkyl having C.sub.3 to
C.sub.6 carbon atoms.
[0111] The present invention also concerns the pharmaceutical
composition and method of treatment of IL-1.beta. mediated disease
states or disorders in a mammal in need of such treatment
comprising the administration of IL-1.beta. inhibitors of formula
(I) as the active agent. These disease states and disorders
include: infectious diseases, such as meningitis and salpingitis;
septic shock, respiratory diseases; inflammatory conditions, such
as arthritis, cholangitis, colitis, encephalitis, endocerolitis,
hepatitis, pancreatitis and reperfusion injury, immune-based
disease, such as hypersensitivity; auto-immune diseases, such as
multiple sclerosis; bone diseases; and certain tumors.
[0112] In the practice of this invention an effective amount of a
compound of the invention or a pharmaceutical composition thereof
is administered to the subject in need of, or desiring, such
treatment. These compounds or compositions may be administered by
any of a variety of routes depending upon the specific end use,
including orally, parenterally (including subcutaneous,
intraarticular, intramuscular and intravenous administration),
rectally, buccally (including sublingually), transdermally or
intranasally. The most suitable route in any given case will depend
upon the use, the particular active ingredient, and the subject
involved. The compound or composition may also be administered by
means of controlled-release, depot implant or injectable
formulations as described more fully herein.
[0113] In general, for the uses as described in the instant
invention, it is expedient to administer the active ingredient in
amounts between about 0.1 and 100 mg/kg body weight, most
preferably from about 0.1 to 30 .mu.g/kg body weight for human
therapy, the active ingredient will be administered preferably in
the range of from about 0.1 to about 20-50 .mu.g/kg/day. This
administration may be accomplished by a single administration, by
distribution over several applications or by slow release in order
to achieve the most effective results. When administered as a
single dose, administration will most preferably be in the range of
from about 0.1 to 10 mg/kg of body weight.
[0114] The exact dose and regimen for administration of these
compounds and compositions will necessarily be dependent upon the
needs of the individual subject being treated, the type of
treatment, and the degree of affliction or need. In general,
parenteral administration requires lower dosage than other methods
of administration which are more dependent upon absorption.
[0115] A further aspect of the present invention relates to
pharmaceutical compositions comprising as an active ingredient a
compound of the present invention in admixture with a
pharmaceutically acceptable, non-toxic carrier. As mentioned above,
such compositions may be prepared for use for parenteral
(subcutaneous, intraarticular, intramuscular or intravenous)
administration, particularly in the form of liquid solutions or
suspensions; for oral or buccal administration, particularly in the
form of tablets or capsules; or intranasally, particularly in the
form of powders, nasal drops or aerosols.
[0116] When administered orally (or rectally) the compounds will
usually be formulated into a unit dosage form such as a tablet,
capsule, suppository or cachet. Such formulations typically include
a solid, semi-solid or liquid carrier or diluent. Exemplary
diluents and vehicles are lactose, dextrose, sucrose, sorbitol,
mannitol, starches, gum acacia, calcium phosphate, mineral oil,
cocoa butter, oil of theobroma, alginates, tragacanth, gelatin,
syrup, methylcellulose, polyoxyethylene sorbitar monolaurate,
methyl hydroxybenzoate, propyl hydroxybenzoate, talc, and magnesium
stearate.
[0117] The compositions may be prepared by any of the methods
well-known in the pharmaceutical art, for example as described in
Remington's Pharmaceutical Sciences, 17th edition, Mack Publishing
Company, Easton, Pa., 1985. Formulations for parentera
administration may contain as common excipients sterile water or
saline, alkylene glycols such as propylene glycol, polyalkylene
glycols such as polyethylene glycol, oils of vegetable origin,
hydrogenated naphthalenes and the like. Examples of vehicles for
parenteral administration include water, aqueous vehicles such as
saline, Ringer's solution, dextrose solution, and Hank's solution
and nonaqueous vehicles such as fixed oils (such as corn,
cottonseed, peanut, and sesame), ethyl oleate, and isopropyl
myristate. Sterile saline is a preferred vehicle and the compounds
are sufficiently water soluble to be made up as a solution for all
foreseeable needs. The vehicle may contain minor amounts of
additives such as substances that enhance solubility, isotonicity,
and chemical stability, e.g., antioxidants, buffers, and
preservatives. For oral administration, the formula can be enhanced
by the addition of bile salts and also by the addition of
acylcarnitines (Am. J. Physiol. 251:332 (1986)). Formulations for
nasal administration may be solid and contain as excipients, for
example, lactose or dextran, or may be aqueous or oily solutions
for administration in the form of nasal drops or metered spray. For
buccal administration typical excipients include sugars, calcium
stearate, magnesium stearate, pregelatinated starch, and the
like.
[0118] When formulated for nasal administration the absorption
across the nasal mucous membrane is enhanced by surfactant acids,
such as for example, glycocholic acid, cholic acid, taurocholic
acid, desoxycholic acid, chenodesoxycholic acid, dehydrocholic
acid, glycodeoxy-cholic acid, and the like (See, B. H. Vickery,
"LHRH and its Analogs-Contraception and Therapeutic Applications",
Pt. 2, B. H. Vickery and J. S. Nester, Eds., MTP Press, Lancaster,
UK, 1987).
DETAILED DESCRIPTION OF THE INVENTION
[0119] Compounds of the present invention are prepared using the
procedure described generally in Schemes I, II and III and in more
detail described in the Examples. 14
[0120] wherein A, R.sub.3 and m are as defined in formula (I) and Z
is benzyloxycarbonyl. 15
[0121] wherein Z, A, R.sub.1, R.sub.3, m and n are as defined in
formula (I). 16
[0122] Methods of Preparation
[0123] The synthesis of the disclosed interleukin enzyme (ICE)
inhibitors was conducted by one of two methods depicted in Schemes
I and II. For inhibitors which contained an N-terminal
benzyloxycarbonyl group ("Z" group), N-benzyloxycarbonyl-L-aspartic
acid mon t-butyl ester or other benzyloxycarbonyl protected
aspartic acid-based peptides (Formula A) were used as starting
materials. The synthesis of the requisite peptides are readily
carried out by a variety of methods known to those practicing in
the art of peptide chemistry. The aspartic acid-based peptide
(Formula A) is reacted with ethyl chloroformate and N-methyl
morpholine in tetrahydrofuran (THF) at low temperature (ca.
-15.degree. C.) for approximately 30 min. This generates a mixed
anhydride in solution thereby activating the free carboxylate
toward nucleophilic attack. Other activating reagents (e.g.
isopropyl chloroformate), solvents (diethyl ether, dioxane), and
tertiary amine bases (diisopropyl ethyl amine, triethyl amine) can
be used in place of the above preferred reagents to form a reactive
carboxylate species. The pre-formed mixed anhydride is treated
(without isolation) with a solution of diazomethane in diethyl
ether. The diazomethane reagent is prepared under standard
conditions from DIAZALD.RTM. using a commercially available
(Aldrich) diazomethane generator. A one to two molar excess of
diazomethane is added and the reaction mixture is warmed from
-15.degree. C. to 25.degree. C. over a 20 min period. During this
time, diazomethane reacts with the mixed anhydride to form an
.alpha.-diazoketone. The .alpha.-diazoketone is not isolated by the
reaction mixture is treated directly with an excess of a 1:1
solution of 48% hydrobromic (HBr) and glacial acetic (HOAc) acids.
The mixture of acids are added dropwise to the .alpha.-diazoketone
and the reaction mixture is subsequently stirred for at least 15
minutes. This treatment with 1:1 48% HBr and glacial HOAc
decomposes the .alpha.-diazoketone to yield the desired
N-benzyloxycarbonyl-L-aspartic acid mono t-butyl ester
.alpha.-bromoketone (Formula B) and nitrogen gas as a by-product.
The bromomethyl ketone is typically isolated as an oil using
standard procedures which are apparent to those skilled in the art.
The .alpha.-bromoketone so obtained is of sufficient purity to be
used in all subsequent reactions. However, the ketone can be
further purified by high pressure liquid chromatography (HPLC), if
analytically pure material is desired.
[0124] The t-butyl ester .alpha.-bromoketone (Formula B) is
subsequently reacted with a variety of phenols, naphthols, and
arylcarboxylic acids. This is conducted by exposing the bromomethyl
ketone to an excess of the phenol or arylcarboxylic acid in
dimethylformamide containing sodium or potassium hydride or
potassium fluoride. The reaction can be conveniently monitored by
thin layer chromatography (TLC) and once the TLC indicates that
displacement of the bromide with the phenol or carboxylate is
completed, the product is isolated using standard procedures. The
desired aspartic acid mono t-butyl ester .alpha.-aryloxymethyl- or
.alpha.-arylacyloxymethyl ketone (Formula C) may be purified by
conventional methods including recrystallization and silica gel
column chromatography.
[0125] The remaining synthetic transformation to generate the ICE
inhibitors is the hydrolysis of the t-butyl ester function. This is
conducted by exposing the t-butyl ester (Formula C) to a 25%
solution of trifluoroacetic acid (TFA) in methylene chloride at
25.degree. C. The de-esterification is typically complete within 3
hrs. Removal of the volatile TFA and organic solvent affords the
aspartic acid (Formula 1). The yield of the reaction is
quantitative in most instances, providing the t-butyl ester
starting material is of high purity. Purification, if required, can
be performed by recrystallization or chromatographic techniques
which are well known to those skilled in the art. The concentration
of TFA may range from 5%-100% and other organic solvents may be
used such as chloroform. Alternatively, a solution of three molar
anhydrous hydrochloric acid in ethyl acetate may be used in place
of the TFA-methylene chloride solution with equal efficiency. the
.sup.1H NMR spectra of these acids of Formula 1 indicate that they
exist in equilibrium as the closed hemiketal form shown in Formula
1A and that the ratio of Fomula 1 versus Formula 1A is solvent
dependent.
[0126] In Scheme II, the synthesis of aryloxy- and
arylacyloxymethyl ketones (Formula 2) which possess an N-terminal
group other than the Z-group are described. The aspartic acid
derivatives of Formula C are the starting material for the
synthesis of inhibitors of Formula 2. First the Z-group is removed
to generate the N-terminal amine (Formula D) under hydrogenolytic
conditions. The reagents and conditions typically used to carry out
the hydrogenolytic removal of the Z-group are hydrogen gas, ambient
temperature conditions and pressure, 5% palladium on carbon as the
catalyst in an alcoholic solvent e.g., methanol, optionally
containing two equivalents of hydrochloric acid. It is not
necessary to purify the intermediate free amine (or the
hydrochloride salt if hydrochloric acid is used in the
hydrogenolysis), though this material needs to be dry and free of
alcohol for the subsequent coupling reaction to proceed in good
yield.
[0127] The N-terminal amine is then condensed with a carboxylic
acid to yield intermediates of Formula E. It is generally necessary
to first activate the acid as an acid chloride or mixed anhydride
and then react it with the free amine (or hydrolchloride salt) in
the presence of an organic base, e.g., N-methylmorpholine.
Alternatively, coupling with acid with the intermediate amine is
conducted using amide coupling reagents/conditions employed in
peptide coupling chemistry ("The Practice of Peptide Synthesis", M.
Bodanszky, Springer-Verlag, NY, 1984; "The Peptides", Vol 1-3, E.
Gross and J. Meienhofer, Eds. Academic Press, NY, 1981). Lastly,
the t-butyl ester in Formula E is removed with trifluoroacetic acid
(as described above) to give the aspartic acid analogs of Formula
2. As in the case of the compounds of Formula 1, the .sup.1H NMR of
components of Formula 2 appear to exist in equilibrium with their
corresponding closed hemiketal counterparts of Formula 2A.
[0128] The phenols, naphthyls and arylcarboxylic acids used in the
reaction with the bromomethyl ketones can be either purchased form
commercial sources or synthesized by adopting known procedures.
Their synthesis would be readily deduced by those skilled in the
art of organic synthesis. By way of example, the preparation of the
2,6-dichloro-3-sulfonamido benzoic acids are presented in Scheme
III. Thus, 2,6-dichlorobenzoic acid (Formula F; available from
Aldrich Chemical Co.) is reacted with chlorosulfonic acid to yield
the intermediate sulfonyl chloride (Formula G). The electrophilic
sulfonyl chloride is reacted with a variety of amines to give the
substituted benzoic acids (Formula 3).
[0129] Intermediate compounds for use in making the final compounds
of the present invention are described in Examples 1-37.
EXAMPLE 1
[0130] N-Benzyloxycarbonyl-L-aspartic acid bromomethyl ketone
.beta.-tert-butyl ester
[0131] To a solution of N-benzyloxycarbonyl L-aspartate
.beta.-tert-butyl ester (Formula A; 10 g, 31 mmol) in 70 ml of
anhydrous THF at -15.degree. C. was added N-methyl morpholine (4.7
ml, 43.4 mol) followed by the dropwise addition of ethyl
chloroformate (3.9 ml, 40.5 mmol). The reaction mixture was stirred
for 30 min at -15.degree. C. and the suspension treated with
diazomethane in ether (160 ml of a 0.4 in solution in ether,
prepared from "DIAZALD.RTM." [Aldrich]) and warmed to room
temperature.
[0132] The bromomethyl ketone was formed in the same pot by cooling
the intermediate diazoketone above followed by the dropwise
addition of a 1:1 solution of 48% hydrobromic acid and glacial
acetic acid (62 ml). After stirring for 15 min the reaction mixture
was poured into a separatory funnel. The aqueous layer was drawn
off and discarded. The remaining organic phase was washed with
water, saturated aqueous NaHCO.sub.3, brine and dried (MgSO.sub.4).
The solvents were removed in vacuo and the title compound so
obtained (m.p. 41-43.degree. C.) was used in the subsequent
displacement reactions without further purification.
EXAMPLE 2
[0133] N-Benzyloxycarbonyl-L-Aspartic Acid
2,6-dichlorobenzoyloxymethyl Ketone .beta.-tert-Butyl Ester
[0134] N-Benzyloxycarbonyl-L-aspartic acid bromomethyl ketone
.beta.-tert-butyl ester (0.30 g; 0.76 mM) was dissolved in 12 ml of
anhydrous DMF. To this solution was added powdered potassium
fluoride (0.11 g; 19 mmol) and 2,6-dichlorobenzoic acid (0.17 g;
0.91 mmol) and the reaction mixture was stirred overnight. The
solution was diluted with Et.sub.2O and washed with water, aqueous
saturated NaHCO.sub.3, brine and dried (MgSO.sub.4). The ketone so
obtained was purified by silica gel chromatography using ethyl
acetate/hexane as the eluting solvent (.sup.1H NMR (CDCl.sub.3)
.delta.7.35 (m, 8H)), 5.90 (d, 2H each), 5.20 (m, 4H), 4.70 (m,
1H), 3.00 and 2.75 (doublet of doublets, 1H each), 1.42 (m,
9H).
[0135] In a similar manner, the following compounds of formula B
were prepared:
EXAMPLE 3
[0136] N-Benzyloxycarbonyl-L-aspartic acid 2,6-difluorophenoxy-
methyl ketone .beta.-tert-butyl ester from
N-benzyloxycarbonyl-L-aspartic acid bromomethyl ketone
.beta.-tert-butyl ester and 2,6-difluorophenol (mp 50-51.degree.
C.)
EXAMPLE 4
[0137] N-Benzyloxycarbonyl-L-aspartic acid 2,6-ditrifluoromethyl
benzyloxymethyl ketone .beta.-tert-butyl ester from
N-benzyloxycarbonyl-L-aspartic acid bromomethyl ketone
.beta.-tert-butyl ester and 2,6-ditrifluoromethyl benzoic acid (mp
62-63.degree. C.)
EXAMPLE 5
[0138] N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichlorophenoxymethyl ketone .beta.-tert-butyl ester from
N-benzyloxycarbonyl-L-aspartic acid bromomethyl ketone
.beta.-tert-butyl ester and 2,6-dichlorophenol.
EXAMPLE 6
[0139] N-Benzyloxycarbonyl-L-aspartic acid
2-fluoro-4-(N-morpholinyl sulfonamido)phenoxymethyl ketone
.beta.-tert-butyl ester from N-benzyloxycarbonyl-L-aspartic acid
bromomethyl ketone .beta.-tert-butyl ester and
2-fluoro-4-(N-morpholinylsulfonamido)phenol.
EXAMPLE 7
[0140] N-Benzyloxycarbonyl-L-aspartic acid
2-chloro-4-(N-thiomorpholinylsu- lfonamido)phenoxymethyl ketone
.beta.-tert-butyl ester from N-benzyloxycarbonyl-L-aspartic acid
bromomethyl ketone .beta.-tert-butyl ester and
2-chloro-4-(N-thiomorpholinyl sulfonamido)phenol.
EXAMPLE 8
[0141] N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichloro-3-(2-N-morpholinyl- ethoxy)benzyloxymethyl ketone
.beta.-tert-butyl ester from N-benzyloxycarbonyl-L-aspartic acid
bromomethyl ketone .beta.-tert-butyl ester and
2,6-dichloro-3-(2-N-morpholinylethoxy) benzoic acid.
EXAMPLE 9
[0142] N-Benzyloxycarbonyl-L-aspartic acid 2,6-dimethoxy
benzoyloxymethyl ketone .beta.-tert-butyl ester from
N-benzyloxycarbonyl-L-aspartic acid bromomethyl ketone
.beta.-tert-butyl ester and 2,6-dimethoxy-benzoic acid.
EXAMPLE 10
[0143] N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichloro-3-(benzyloxy) benzoyloxymethyl ketone
.beta.-tert-butyl ester from N-benzyloxycarbonyl-L-aspartic acid
bromomethyl ketone .beta.-tert-butyl ester and
2,6-dichloro-3-(benzoyloxy)benzoic acid.
EXAMPLE 11
[0144] N-Benzyloxycarbonyl-L-aspartic acid 2-acetamido-6-chloro
benzoyloxymethyl ketone .beta.-tert-butyl ester from
N-benzyloxycarbonyl-L-aspartic acid bromomethyl ketone
.beta.-tert-butyl ester and 2-acetamido-6-chlorobenzoic acid.
EXAMPLE 12
[0145] N-Benzyloxycarbonyl-L-aspartic acid
2,6-difluorobenzoyloxy-methyl ketone .beta.-tert-butyl ester from
N-benzyloxycarbonyl-L-aspartic acid bromomethyl ketone
.beta.-tert-butyl ester and 2,6-difluorobenzoic acid.
EXAMPLE 13
[0146] N-Benzyloxycarbonyl-L-aspartic acid
3-(N-butylsulfonamido)-2,6-dich- lorobenzoyloxymethyl ketone
.beta.-tert-butyl ester from N-benzyloxycarbonyl-L-aspartic acid
bromomethyl ketone .beta.-tert-butyl ester and
3-(N-butylsulfonamido)-2,6-dichlorobenzoic acid.
EXAMPLE 14
[0147] N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichloro-3-sulfonamido benzoyloxymethyl ketone
.beta.-tert-butyl ester from N-benzyloxycarbonyl-L-aspartic acid
bromomethyl ketone .beta.-tert-butyl ester and
2,6-dichloro-3-sulfonamidobenzoic acid.
EXAMPLE 15
[0148] N-Benzyloxycarbonyl-L-aspartic
acid-3-(N-benzylsulfonamido)-2,6-dic- hlorobenzoyloxymethyl ketone
.beta.-tert-butyl ester from N-benzyloxycarbonyl-L-aspartic acid
bromomethyl ketone .beta.-tert-butyl ester and
3-(N-benzylsulfonamido)benzoic acid.
EXAMPLE 16
[0149] N-Benzyloxycarbonyl-L-aspartic acid
3-(N-[2-aminoacetamidoyl]sulfon-
amido)-2,6-dichlorobenzoyloxymethyl ketone .beta.-tert-butyl ester
from N-benzyloxycarbonyl-L-aspartic acid bromomethyl ketone
.beta.-tert-butyl ester and
3-(N-[2-aminoacetamidoyl]sulfonamido)-2,6-dichlorobenzoic acid.
EXAMPLE 17
[0150] N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichloro-3-(N-morpholinylsu- lfonamido)benzoyloxymethyl ketone
.beta.-tert-butyl ester from N-benzyloxycarbonyl-L-aspartic acid
bromomethyl ketone .beta.-tert-butyl ester and
2,6-dichloro-3-N-morpholinyl sulfonamido)benzoic acid.
EXAMPLE 18
[0151] N-Methoxycarbonyl-L-Alanine-L-Aspartic Acid
2,6-dichloro-benzoyloxy- methyl Ketone .beta.-tert-Butyl Ester and
other Compounds of Formula E
[0152] Part A:
[0153] N-benzyloxycarbonyl-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone .beta.-tert-butyl ester (1.02
g, 2 mmol) was dissolved in absolute ethanol (100 ml) containing 6
N aqueous HCl (0.67 ml, 4 mmol). 10% Palladium on carbon (96 mg)
was added and the reaction mixture was stirred under an ambient
atmosphere of hydrogen gas for approximately 1 hour (thin layer
chromotography [5% MeOH--CH.sub.2Cl.sub.2] indicated the
disappearance of starting material). The solution was filtered and
the solvent was removed in vacuo to give L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone .beta.-tert butyl ester-HCl
salt (Formula D) which was used immediately in the subsequent
reaction described in Part B.
[0154] Part B:
[0155] A solution of N-methoxycarbonyl-L-alanine (301 mg, 2.05
mmol) in CH.sub.2Cl.sub.2 (10 ml) was cooled to -20.degree. C. and
isobutylchloroformate (0.28 ml, 2.05 mmol) and N-methylmorpholine
(0.23 ml, 2.05 mmol) were added sequentially. The reaction mixture
was stirred for 15 minutes and a solution of aspartic acid
2,6-dichlorobenzoyl methyl ketone .beta.-tert-butyl ester-HCl salt
(prepared in Part A above) followed by a second addition of
N-methyl morpholine (0.23 ml, 2.05 mmol).
[0156] The reaction mixture was stirred for 30 minutes and was then
diluted with EtOAc, washed with water, aqueous saturated
NaHCO.sub.3, brine and dried (MgSO.sub.4). The solvents were
removed in vacuo and the product purified by silica gel
chromatography using 40% EtOAc--hexane as eluent to give
N-methoxycarbonyl-L-alanine-L-aspartic acid 2,6-dichlorobenzoyl
methyl ketone .beta.-tert ester (0.72 g; 80%).
[0157] In a similar fashion the following compounds of Formula E
were prepared:
EXAMPLE 19
[0158] N-(2-Thienyl)carbonyl-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone .beta.-tert-butyl ester from
N-benzyloxycarbonyl-L-aspartic acid 2,6-dichlorobenzoyloxymethyl
ketone .beta.-tert-butyl ester and 2-thiophene carboxylic acid.
EXAMPLE 20
[0159] N-Methoxycarbonyl glycine-L-aspartic acid
2,6-dichlorobenzoyloxymet- hyl ketone .beta.-tert-butyl ester from
N-benzyloxycarbonyl-L-aspartic acid 2,6-dichlorobenzoyloxymethyl
ketone .beta.-tert-butyl ester and N-methoxycarbonyl glycine.
EXAMPLE 21
[0160] N-Methoxycarbonyl-L-phenylalanine-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone .beta.-tert-butyl ester from
N-benzyloxycarbonyl-L-aspartic acid 2,6-dichlorobenzoyloxymethyl
ketone .beta.-tert-butyl ester and N-methoxycarbonyl-L-phenyl
alanine.
EXAMPLE 22
[0161] N-Methoxycarbonyl L-.beta.-(2-thienyl)alanine-L-aspartic
acid 2,6-dichlorobenzoyloxymethyl ketone .beta.-tert-butyl ester
from N-benzyloxycarbonyl-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone .beta.-tert-butyl ester and
N-methoxycarbonyl-L-.beta.-(2-thienyl)alaine.
EXAMPLE 23
[0162] N-Methoxycarbonyl-L-valine-L-aspartic acid
2,6-dichlorobenzoyloxyme- thyl ketone .beta.-tert-butyl ester from
N-benzyloxycarbonyl-L-aspartic acid 2,6-dichlorobenzoyloxymethyl
ketone .beta.-tert-butyl ester and N-methoxycarbonyl-L-valine.
EXAMPLE 24
[0163] N-Methoxycarbonyl-L-histidine-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone .beta.-tert-butyl ester from
N-benzyloxycarbonyl-L-aspartic acid 2,6-dichlorobenzoyloxymethyl
ketone .beta.-tert-butyl ester and
N-methoxycarbonyl-L-histidine.
EXAMPLE 25
[0164] N-Benzyloxycarbonyl-L-alanine-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone .beta.-tert-butyl ester from
N-benzyloxycarbonyl-L-aspartic acid 2,6-dichlorobenzoyloxymethyl
ketone .beta.-tert-butyl ester and
N-benzyloxycarbonyl-L-valine.
EXAMPLE 26
[0165] N-Benzyloxycarbonyl-L-alanine-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone .beta.-tert-butyl ester from
N-benzyloxycarbonyl-L-aspartic acid 2,6-dichlorobenzoyloxymethyl
ketone .beta.-tert-butyl ester and
N-benzyloxycarbonyl-L-alanine.
EXAMPLE 27
[0166] i) Benzyloxycarbonyl-L-valine-L-alanine-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone .beta.-tert-butyl ester from
N-benzyloxycarbonyl-L-aspartic acid 2,6-dichlorobenzoyloxymethyl
ketone .beta.-tert-butyl ester and
N-benzyloxycarbonyl-L-valine-L-alanine
EXAMPLE 28
[0167] N-2-Furoyl-L-Aspartic Acid 2,6-Dichlorobenzoyloxymethyl
Ketone .beta.-tert-Butyl Ester
[0168] Part A:
[0169] N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone .beta.-tert-butyl ester (1.02
g, 2 mmol) was dissolved in absolute ethanol (100 ml) containing 6
N aqueous HCl (0.67 ml, 4 mmol). 10% Palladium on carbon (96 mg)
was added and the reaction mixture was stirred under an ambient
atmosphere of hydrogen gas for approximately 1 hour (thin layer
chromotography [15% MeOH--CH.sub.2Cl.sub.2] indicated the
disappearance of starting material). The solution was filtered and
the solvent was removed in vacuo to give L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone .beta.-tert-ester-HCl salt
(Formula D) which was used immediately in the subsequent reaction
described in Part B.
[0170] Part B:
[0171] To a solution of L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone .beta.-tert-butyl ester-HCl
salt (2.0 mmol, prepared in Part A above) in CH.sub.2Cl.sub.2 (10
ml) at 0.degree. C. was added 2-furoyl chloride (0.21 ml, 2.05
mmol). N-methylmorpholine (0.25 ml; 2.10 mmol) was then added and
the reaction mixture stirred for 1 hour as it slowly was allowed to
warm to room temperature. The solution was diluted with EtOAc,
washed with water, saturated aqueous NaHCO.sub.3, brine and dried
(MgSO.sub.4). The solvents were removed in vacuo. The product was
purified by silica gel chromatography using 30% EtOAc-hexane as
eluent to give N-2-furoyl-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone-tert-butyl ester (mp
73-74.degree. C.).
[0172] In a similar fashion the following compounds of Formula E
were prepared:
EXAMPLE 29
[0173] N-2-Furonylcarbonyl-L-aspartic acid
2,6-dichloro-3-(N-morpholinylsu- lfonamido) benzoyloxymethyl ketone
.beta.-tert-butyl ester from N-benzyloxycarbonyl-L-aspartic acid
2,6-dichloro-3-(N-morpholinylsulfonam- ido) benzoyloxymethyl ketone
.beta.-tert-butyl ester and 2-furoic acid chloride.
EXAMPLE 30
[0174] N-(3-Phenylpropionyl)-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone .beta.-tert-butyl ester from
N-benzyloxycarbonyl-L-aspartic acid 2,6-dichlorobenzoyloxymethyl
ketone .beta.-tert-butyl ester and 3-phenylpropionyl chloride.
EXAMPLE 31
[0175] N-Methoxycarbonyl-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone .beta.-tert-butyl ester from
N-benzyloxycarbonyl-L-aspartic acid 2,6-dichlorobenzoyloxymethyl
ketone .beta.-tert-butyl ester and methyl chloroformate.
EXAMPLE 32
[0176] N-(N,N-4-Dimethylaminomethyl)benzoyl-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone .beta.-tert-butyl ester (m.p.
63-65.degree. C.) from 2,6-dichlorobenzoyloxymethyl ketone
.beta.-tert-butyl ester and 4-(N,N-dimethylaminomethyl)benzoyl
chloride.
EXAMPLE 33
[0177] 3-(N-Butylsulfonamidoyl)-2,6-dichlorobenzoic acid and Other
Compound of Formula 3
[0178] Part A:
[0179] Under an atmosphere of nitrogen gas, a reaction vessel was
charged with 2,6-dichlorobenzoic acid (10 g, 53.55 mmol) (Formula
F) and chlorosulfonic acid (3 ml, 472 mmol). The reaction mixture
was refluxed for 1 hour and cooled to 10.degree. C. The contents of
the reaction vessel were poured slowly into 3 L of ice water. The
white solid which precipitated was collected by filtration and
dried in vacuo (10 mm) at 35.degree. C. for 48 hours to give
3-(chlorosulfonyl)-2,6-dichlorobenzoic acid (Formula G) (9.2 g, 61%
yield).
[0180] Part B:
[0181] 3-(Chlorosulfonyl)-2,6-dichlorobenzoic acid (1.5 g; 5.2
mmol) was dissolved in anhydrous toluene (35 ml) to which was added
powdered K.sub.2CO.sub.3 (1.44 g: 10.4 mmol) and n-butylamine (1.0
ml, 10.4 mmol). The reaction mixture was stirred at 25.degree. C.
for 12 hours. The solution was diluted slowly with 1 M ethereal HCl
(20 ml) and was then stirred for 30 minutes. The solution was
filtered and the resulting filtrate was evaporated to dryness to
give crude product. Further purification of the material by silica
gel chromatography using EtOAc as the eluent provided
3-(N-butylsulfonamidoyl)-2,6-dichlorobenzoic acid (Formula 3) (1.43
g, 85%. .sup.1H NMR (DMSO) .delta.8.11 (t, 1H), 7.98 and 7.71
(doublets, 1H each), 2.75 (m, 2H), 1.55 (m, 2H), 1.32 (m 2H), 0.87
(t, 3H).
[0182] In a similar manner, the following compounds were
prepared:
EXAMPLE 34
[0183] 2,6-Dichloro-3-sulfonamidoylbenzoic acid (.sup.1H NMR (DMSO)
.delta.8.11 (t, 1H), 7.42 and 7.15 (doublets, 1H each), 7.26 (d,
2H) from 3-chlorosulfonyl-2,6-dichlorobenzoic acid and 40% aqueous
ammonium hydroxide.
EXAMPLE 35
[0184] 3-(N-Benzylsulfonamidoyl)-2,6-dichlorobenzoic acid (.sup.1H
NMR (DMSO) .delta.8.70 (t, 1H), 7.90 and 7.65 (doublets, 1H each),
7.25 (m, 5H), 4.15 (m, 2H) from
3-chlorosulfonyl-2,6-dichlorobenzoic acid and benzyl amine.
EXAMPLE 36
[0185] 3-(N-[2-Aminoacetamido]sulfonamidoly)-2,6-dichlorobenzoic
acid from 3-chlorosulfonyl-2,6-dichlorobenzoic acid and glycinamide
(m. p. 210-213.degree. C.
EXAMPLE 37
[0186] 3-(N-Morpholino)sulfonamidoyl)-2,6-dichlorobenzoic acid from
3-chlorosulfonyl-2,6-dichlorobenzoic acid and morpholine.
EXAMPLE 38
[0187] N-Benzyloxycarbonyl-L-Aspartic Acid
2,6-Dichlorobenzoyloxymethyl Ketone and Other Compounds of Formula
I
[0188] A solution of .beta.-tert-butyl ester of
N-benzyloxycarbonyl-L-aspa- rtic acid 2,6-dichlorobenzoyloxymethyl
ketone (Example 2) in methylene chloride containing 25% v/v
trifluoroacetic acid (20 ml) was stirred for 2 hours at 0.degree.
C. The solvent was removed in vacuo and the residue azeotroped
three times with methylene chloride to give analytically pure
N-benzyloxycarbonyl-L-aspartic acid 2,6-dichlorobenzoyloxymethyl
ketone (high resolution mass spectrum for
C.sub.20H.sub.17Cl.sub.2NO.sub.7 found 453.1572).
[0189] In a similar fashion, the following compounds of Formulas 1
and 2 were prepared:
EXAMPLE 39
[0190] N-Benzyloxycarbonyl-L-aspartic acid
2,6-difluorophenoxymethyl ketone (high resolution mass spectrum for
C.sub.19H.sub.17F.sub.2NO.sub.6 found 393.3562) from the
.beta.-tert-butyl ester of Example 3.
EXAMPLE 40
[0191] N-Benzyloxycarbonyl-L-aspartic acid 2,6-ditrifluoromethyl
benzoyloxymethyl ketone (high resolution mass spectrum for
C.sub.22H.sub.170.sub.7F.sub.6 found 521.1452) from the
.beta.-tert-butyl ester of Example 4.
EXAMPLE 41
[0192] N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichlorophenoxymethyl ketone (mass spectrum m/z 426 (M+H) from
the .beta.-tert-butyl ester of Example 5.
EXAMPLE 42
[0193] N-Benzyloxycarbonyl-L-aspartic acid
2-fluoro-4-(N-morpholinyl sulfonamido)phenoxymethyl ketone (m.p.
65-66.degree. C.) from the .beta.-tert-butyl ester of Example
6.
EXAMPLE 43
[0194] N-Benzyloxycarbonyl-L-aspartic acid
2-chloro-4-(N-thiomorpholinyl-s- ulfonamido)phenoxymethyl ketone
(m.p. 180-181.degree. C.) from the .beta.-tert-butyl ester of
Example 7.
EXAMPLE 44
[0195] N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichloro-3-(2-N-morpholinyl- ethoxy)benzoyloxymethyl ketone
(high resolution mass spectrum for
C.sub.26H.sub.29O.sub.9N.sub.2Cl.sub.2 found 583.1245 ) from the
.beta.-tert-butyl ester of Example 8.
EXAMPLE 45
[0196] N-Benzyloxycarbonyl-L-aspartic acid 2,6-dimethoxybenzoyloxy
methyl ketone (high resolution mass spectrum for
C.sub.22H.sub.24O.sub.9N found 446.1430 ) from the
.beta.-tert-butyl ester of Example 9.
EXAMPLE 46
[0197] N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichloro-3-(benzyloxy) benzoyloxymethyl ketone (high resolution
mass spectrum for C.sub.27H.sub.24O.sub.8NCl.sub.2 found 560.0865 )
from the .beta.-tert-butyl ester of Example 10.
EXAMPLE 47
[0198] N-Benzyloxycarbonyl-L-aspartic acid
2-acetamido-6-chlorobenzoyloxym- ethyl ketone (high resolution mass
spectrum for C.sub.22H.sub.22O.sub.8N.s- ub.2Cl.sub.2 found
477.1044) from the .beta.-tert-butyl ester of Example 11.
EXAMPLE 48
[0199] N-Benzyloxycarbonyl-L-aspartic acid
2,6-difluorobenzoyloxymethyl ketone (high resolution mass spectrum
for C.sub.20H.sub.18O.sub.7NF.sub.2 found 422.1046) from the
.beta.-tert-butyl ester of Example 12.
EXAMPLE 49
[0200] N-Benzyloxycarbonyl-L-aspartic acid
3-(N-butylsulfonamido)-2,6-dich- lorobenzoyloxymethyl ketone (m.p.
48-50.degree. C.) from the .beta.-tert-butyl ester of Example
13.
EXAMPLE 50
[0201] N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichloro-3-sulfonamido benzoyloxymethyl ketone (m.p.
44-46.degree. C.) from the .beta.-tert-butyl ester of Example
14.
EXAMPLE 51
[0202] N-Benzyloxycarbonyl-L-aspartic acid
3-(N-benzylsulfonamido)-2,6-dic- hlorobenzoyloxymethyl ketone (m.p.
66-68.degree. C.) from the .beta.-tert-butyl ester of Example
15.
EXAMPLE 52
[0203] N-Benzyloxycarbonyl-L-aspartic acid
3-(N-[2-aminoacetamidoyl]sulfon-
amido)-2,6-dichlorobenzoyloxymethyl ketone (m.p. 54-56.degree. C.)
from the .beta.-tert-butyl ester of Example 16.
EXAMPLE 53
[0204] N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichloro-3-(N-morpholinylsu- lfonamido)benzoyloxymethyl ketone
(high resolution mass spectrum for
C.sub.24H.sub.25O.sub.10N.sub.2Cl.sub.2 found 603.0594) from the
.beta.-tert-butyl ester of Example 17.
EXAMPLE 54
[0205] N-Methoxycarbonyl-L-alanine-L-aspartic acid
2,6-dichlorobenzoyloxym- ethyl ketone (Anal. calc. for
C.sub.17H.sub.18O.sub.8Cl.sub.2N.sub.2: C, 45.45; H, 4.04; N, 6.24.
Found: C, 45.20; H, 4.06; N, 5.98) from the .beta.-tert-butyl ester
of Example 18.
EXAMPLE 55
[0206] N-(2-thienyl)carbonyl-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone (mass spectrum m/z 430 (M+))
from the .beta.-tert-butyl ester of Example 19.
EXAMPLE 56
[0207] N-Methoxycarbonyl-glycine-L-aspartic acid
2,6-dichlorobenzoyloxymet- hyl ketone (Anal. calc. for
C.sub.16H.sub.16O.sub.8Cl.sub.2N.sub.2: C, 44.16; H, 3.17; N, 6.44.
Found: C, 44.24; H, 3.15; N, 6.12) from the .beta.-tert-butyl ester
of Example 20.
EXAMPLE 57
[0208] N-Methoxycarbonyl-L-phenylalanine-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone (Anal. calc. for
C.sub.23H.sub.22O.sub.8Cl.sub.2N.sub.2: C, 52.59; H, 4.22; N, 5.33.
Found: C, 52.98; H, 4.38; N, 5.21) from the .beta.-tert-butyl ester
of Example 21.
EXAMPLE 58
[0209] N-Methoxycarbonyl-L-.beta.-(2-thienyl)alanine-L-aspartic
acid 2,6-di-chlorobenzoyloxymethyl ketone (mass spectrum m/z 531
(M+)) from the .beta.-tert-butyl ester of Example 22.
EXAMPLE 59
[0210] N-Methoxycarbonyl-L-valine-L-aspartic acid
2,6-dichlorobenzoyloxyme- thyl ketone (m.p. 119-120.degree. C.)
from the .beta.-tert-butyl ester of Example 23.
EXAMPLE 60
[0211] N-Methoxycarbonyl-L-histidine-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone (Anal. calc. for
C.sub.22H.sub.21O.sub.10F.sub.3Cl.sub.2N.sub.4: C, 41.99; H, 3.36;
N, 8.90. Found: C, 42.08; H, 3.48; N, 8.67; mass spectrum m/z 515
(M+)) from the .beta.-tert-butyl ester of Example 24.
EXAMPLE 61
[0212] N-Benzyloxycarbonyl-L-valine-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone (Anal. calc. for
C.sub.25H.sub.26O.sub.8Cl.sub.2N.sub.2: C, 54.26; H, 4.47; N, 5.06.
Found: C, 54.06; H, 4.74; N, 4.91) from the .beta.-tert-butyl ester
of Example 25.
EXAMPLE 62
[0213] N-Benzyloxycarbonyl-L-alanine-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone (mass spectrum m/z 525 (M+))
from the .beta.-tert-butyl ester of Example 26.
EXAMPLE 63
[0214] N-Benzyloxycarbonyl-L-valine-L-alanine-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone (Anal. calc. for
C.sub.28H.sub.31O.sub.9Cl.sub.2N.sub.3: C, 53.85; H, 5.00; N, 6.73.
Found: C, 54.00; H, 5.04; N, 6.66) from the .beta.-tert-butyl ester
of Example 27.
EXAMPLE 64
[0215] N-(2-Furonyl)carbonyl-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone (mass spectrum m/z 414 (M+))
from the .beta.-tert-butyl ester of Example 28.
EXAMPLE 65
[0216] N-(2-Furonyl)carbonyl-L-aspartic acid
2,6-dichloro-3-(N-morpholinyl- sulfonamido)benzoyloxymethyl ketone
(mass spectrum m/z 563 (M+)) from the .beta.-tert-butyl ester of
Example 29.
EXAMPLE 66
[0217] N-(3-Phenylpropionyl)-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone (.sup.1H NMR (CDCl.sub.3)
.delta.7.40 (m, 9H), 5.05 (2xdd, 4H), 4,70 (m, 1H), 2.85 (m, 2H),
2.65 (dd, 1H), 2.60 (dd, 1H), 2.50 (m,2H) from the
.beta.-tert-butyl ester of Example 30.
EXAMPLE 67
[0218] N-Methoxycarbonyl-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone (1H NMR (DMSO) .delta.7.60 (m,
6H), 5.24 (m, 4H), 4.51 (m, 1H), 3.58 (s, 3H), 2.75 (dd, 1H), 2.55
(dd, 1H) from the .beta.-tert-butyl ester of Example 31.
EXAMPLE 68
[0219] N-(4-N,N-dimethylaminomethyl)benzoyl-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone (m.p. 55-57.degree. C.) from
the .beta.-tert-butyl ester of Example 32.
EXAMPLE 69
[0220] N-Benzyloxycarbonyl-D-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone (high resolution mass spectrum
for C.sub.20H.sub.17C.sub.12NO.sub.- 7, found 453.1583) from
N-benzyloxycarbonyl-D-aspartic acid-.beta.-tert-butyl ester and
2,6-dichlorobenzoic acid using the procedures described in Examples
1, 2 and 38.
EXAMPLE 70
[0221] N-(2-[2,6-dichlorobenzoyloxy])acetyl-L-aspartic acid
2,6-dichlorobenzoyloxymethyl ketone (mass spectrum m/z 551
(M.sup.+) from N-Benzyloxycarbonyl-L-aspartic acid
2,6-dichlorobenzyloxymethyl ketone and
2-(2,6-dichlorobenzoyloxy)acetic acid using the procedures
described in Examples 18 and 38.
EXAMPLE 71
[0222] N-Benzyloxycarbonyl-L-valine-L-aspartic acid
4-(N,N-diethylsulfonamido)-2,3,5,6-tetrafluorophenoxymethyl ketone
(mass spectrum m/z 664 (M+H) from N-Benzyloxycarbonyl-L-aspartic
acid bromomethyl ketone .beta.-tert-butyl ester,
N-benzyloxycarbonyl-L-valine and
4-(N,N-diethylsulfonamido)-2,3,5,6-tetrafluorophenol using the
procedures described in Examples 2, 18 and 38. The
4-(N,N-diethyl-2,3,5,6-tetrafluorophenol was prepared by reacting
2,3,5,6-tetrafluorophenol with chlorosulfonic acid followed by
reaction with diethylamine, analogous to the procedure described in
Scheme III, and Example 33.
[0223] Compounds of the present invention were tested for
IL-1.beta. protease inhibition activity according to the following
protocol:
[0224] Partially purified IL-1.beta. protease is stored at
-80.degree. C., thawed on ice, and preincubated for 10 minutes at
37.degree. C. with 2.5 mM dithiothreitol in a buffer solution
containing 10 mM Tris-HCl (pH 8.0) and 25% (v/w) glycerol.
Inhibitors are prepared as stock solutions in dimethyl sulfoxide
(DMSO). The protease is preincubated with inhibitor in a volume of
20 .mu.l in a 1.5 ml polypropylene microcentrifuge tube for 15
minutes at 37.degree. C. The volume of compound added to the assay
is adjusted to yield a DMSO concentration in the preincubation of
<15% (v/v). The enzyme assay is then initiated by the addition
of substrate (TRITC-AYVHDAPVRS-NH2) to yield a final concentration
of 67 .mu.M in a final volume of 30 .mu.L. The reaction are carried
out for 60 minutes at 37.degree. C. in the dark and are terminated
by the addition of 10 .mu.l of 10% trifluoroacetic acid (TFA).
Following the addition of 115 .mu.l of 0.1% TFA, the samples are
analyzed by high pressure liquid chromatography using a reverse
phase (C18) column and elution with an acetonitrile/water/TFA
gradient. Substrate and product are monitored by their absorbance
at 550 nm and elute at 4.2 and 5.2 minutes, respectively.
1TABLE I Example No. Name of Compound IC.sub.50.mu.m 38
N-Benzyloxycarbonyl-L-aspartic acid 0.05
2,6-dichlorobenzoyloxymethyl ketone 40
N-Benzyloxycarbonyl-L-aspartic acid 0.10 2,6-ditrifluoromethylben-
zoyloxymethyl ketone 41 N-BenzyloxycarbonykL-aspartic acid 0.10
2,6-dichlorophenoxymethyl ketone 42 N-Benzyloxycarbonyl-L-aspartic
acid 0.32 2 fluoro-4-(N-morpholinyl sulfonamido)phenoxymethyl
ketone 49 N-Benzyloxycarbonyl-L-aspartic acid 3-(N- 0.09
butylsulfonamido)-2,6-dichflorobenzoyloxymethyl ketone 52
N-BenzyloxycarbonykL-aspartic acid 3- 0.06
(N-[2-aminoacetamidoyl]sulfonamido)- 2,6-dichlorobenzoyloxymethyl
ketone 53 N-Benzyloxycarbonyl-L-aspartic acid 0.09
2,6-dichloro-3-(N-morpholinylsulfonamido)- benzoyloxymethyl ketone
54 N-Methoxycarbonyl-L-alanine-L-aspartic acid 0.06
2,6-dichlorobenzoyloxymethyl ketone 57 N-Methoxycarbonyl-L-phenyla-
lanine-L-aspartic 0.07 acid 2,6-dichlorobenzoyloxymethyl ketone 64
N-(2-furonyl)carbonyl-L-aspartic acid 2,6- 0.14
dichlorobenzoyloxymethyl ketone 67 N-Methoxycarbonyl-L-aspartic
acid 0.08 2,6-dichlorobenzoyloxymethyl ketone 68
N-(4-(N,N-dimethylaminomethyl)benzoyl-L-aspartic 0.3 acid
2,6-dichlorobenzoyloxymethyl ketone 70 N-(2-[2,6-dichlorobenzoylox-
y])acetyl-L-aspartic 0.2 acid 2,6-dichlorobenzoyloxymethyl
ketone
* * * * *