U.S. patent application number 15/235351 was filed with the patent office on 2016-12-01 for fluorinated epoxyketone-based compounds and uses thereof as proteasome inhibitors.
This patent application is currently assigned to Trillium Therapeutics Inc.. The applicant listed for this patent is Trillium Therapeutics Inc.. Invention is credited to Peter Dove, Abdelmalik Slassi.
Application Number | 20160347792 15/235351 |
Document ID | / |
Family ID | 50101155 |
Filed Date | 2016-12-01 |
United States Patent
Application |
20160347792 |
Kind Code |
A1 |
Slassi; Abdelmalik ; et
al. |
December 1, 2016 |
FLUORINATED EPOXYKETONE-BASED COMPOUNDS AND USES THEREOF AS
PROTEASOME INHIBITORS
Abstract
The present application relates to novel fluorinated
epoxyketone-based compounds, compositions comprising these
compounds and their use, in particular for the treatment of
diseases, disorders or conditions mediated by proteasome
inhibition. In particular, the present application includes
compounds of Formula I, and compositions and uses thereof:
##STR00001##
Inventors: |
Slassi; Abdelmalik;
(Mississauga, CA) ; Dove; Peter; (Burlington,
CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Trillium Therapeutics Inc. |
Mississauga |
|
CA |
|
|
Assignee: |
Trillium Therapeutics Inc.
Mississauga
CA
|
Family ID: |
50101155 |
Appl. No.: |
15/235351 |
Filed: |
August 12, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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14420817 |
Feb 10, 2015 |
9441012 |
|
|
PCT/CA2013/050620 |
Aug 13, 2013 |
|
|
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15235351 |
|
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61682836 |
Aug 14, 2012 |
|
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 27/02 20180101;
A61P 19/02 20180101; A61P 27/12 20180101; A61P 1/04 20180101; A61P
11/06 20180101; A61P 29/00 20180101; A61P 3/04 20180101; A61P 35/02
20180101; A61P 9/10 20180101; C07K 5/0827 20130101; A61P 17/06
20180101; A61P 31/18 20180101; A61P 19/10 20180101; A61P 27/06
20180101; A61P 37/02 20180101; A61P 37/06 20180101; A61K 38/06
20130101; C07K 5/06043 20130101; A61P 1/16 20180101; C07K 5/06078
20130101; A61P 37/08 20180101; A61P 43/00 20180101; A61P 25/28
20180101; A61P 25/16 20180101; A61P 9/00 20180101; C07K 5/06008
20130101; A61P 31/04 20180101; A61P 3/10 20180101; A61P 35/00
20180101; A61P 5/14 20180101; A61P 21/00 20180101; A61P 21/02
20180101; A61P 31/12 20180101; A61P 25/14 20180101; A61P 17/00
20180101; A61K 38/00 20130101 |
International
Class: |
C07K 5/08 20060101
C07K005/08 |
Claims
1. A compound of Formula I, or a pharmaceutically acceptable salt
or solvate thereof: ##STR00114## wherein: R.sup.1 is selected from
the group consisting of C.sub.1-10alkyl, C.sub.2-10alkenyl,
C.sub.2-10alkynyl, C.sub.1-10haloalkyl, C.sub.1-10cyanoalkyl,
C.sub.1-10alkoxy, C.sub.2-10alkenyloxy, C.sub.2-10alkynyloxy,
C.sub.3-10cycloalkyl, heterocycloalkyl, aryl, heteroaryl,
C.sub.1-6alkylene-O--C.sub.1-6alkyl,
C.sub.1-6alkylene-O--C.sub.1-6haloalkyl,
C.sub.2-6alkenylene-O--C.sub.1-6haloalkyl,
C.sub.2-6alkynylene-O--C.sub.1-6haloalkyl,
C.sub.1-6alkylene-C.sub.3-8cycloalkyl,
C.sub.1-6alkylene-heterocycloalkyl, C.sub.1-6alkylene-aryl,
C.sub.1-6alkylene-heteroaryl, C(O)R.sup.7, OC(O)R.sup.7,
C(O)OR.sup.7, C.sub.1-6alkylene-O--R.sup.7,
C.sub.1-6alkylene-C(O)R.sup.7, C.sub.1-6alkylene-O--C(O)R.sup.7,
C.sub.1-6alkylene-C(O)OR.sup.7, C.sub.1-6alkylene-O--C(O)OR.sup.7,
C.sub.1-6alkylene-NR.sup.7R.sup.8,
C.sub.1-6alkylene-C(O)NR.sup.7R.sup.8,
C.sub.1-6alkylene-NR.sup.7C(O)R.sup.8,
C.sub.1-6alkylene-NR.sup.7C(O)NR.sup.7R.sup.8,
C.sub.1-6alkylene-S--R.sup.7, C.sub.1-6alkylene-S(O)R.sup.7,
C.sub.1-6alkylene-SO.sub.2R.sup.7,
C.sub.1-6alkylene-SO.sub.2NR.sup.7R.sup.8,
C.sub.1-6alkylene-NR.sup.7SO.sub.2R.sup.8,
C.sub.1-6alkylene-NR.sup.7SO.sub.2NR.sup.7R.sup.8,
C(O)NR.sup.7R.sup.8 and C.sub.1-6alkylene-NR.sup.7C(O)OR.sup.8,
wherein any cyclic moiety is optionally substituted with
C.sub.1-4alkyl and/or is optionally fused to a further cyclic
moiety; X is absent or is selected from the group consisting of O,
NH, NC.sub.1-6alkyl, S, S(O), SO.sub.2, C(O), C.sub.1-6alkylene,
C.sub.2-6alkenylene, C.sub.2-6alkynylene, C.sub.1-6haloalkylene,
C.sub.3-8cycloalkylene, heterocycloalkylene, arylene and
heteroarylene, or X is a combination of two or three of O, NH,
NC.sub.1-6alkyl, S, S(O), SO.sub.2, C.sub.1-6alkylene,
C.sub.2-6alkenylene, C.sub.2-6alkynylene, C.sub.1-6haloalkylene,
C.sub.3-8cycloalkylene, heterocycloalkylene, arylene or
heteroarylene, bonded together in a linear fashion, provided that
two or three of O, NH, NC.sub.1-6alkyl, S, S(O) and SO.sub.2 and
not bonded directly to each other; R.sup.2, R.sup.3 and R.sup.4 are
each independently selected from the group consisting of
C.sub.1-10alkyl, C.sub.2-10alkenyl, C.sub.2-10alkynyl,
C.sub.1-10haloalkyl, C.sub.1-10cyanoalkyl, C.sub.1-10alkoxy,
C.sub.2-10alkenyloxy, C.sub.2-10alkynyloxy, C.sub.3-10cycloalkyl,
heterocycloalkyl, aryl, heteroaryl,
C.sub.1-6alkylene-O--C.sub.1-6alkyl,
C.sub.1-6alkylene-O--C.sub.1-6haloalkyl,
C.sub.2-6alkenylene-O--C.sub.1-6haloalkyl,
C.sub.2-6alkynylene-O--C.sub.1-6haloalkyl,
C.sub.1-6alkylene-C.sub.3-8cycloalkyl,
C.sub.1-6alkylene-heterocycloalkyl, C.sub.1-6alkylene-aryl,
C.sub.1-6alkylene-heteroaryl, C(O)R.sup.7, OC(O)R.sup.7,
C(O)OR.sup.7, C.sub.1-6alkylene-O--R.sup.7,
C.sub.1-6alkylene-C(O)R.sup.7, C.sub.1-6alkylene-O--C(O)R.sup.7,
C.sub.1-6alkylene-C(O)OR.sup.7, C.sub.1-6alkylene-O--C(O)OR.sup.7,
C.sub.1-6alkylene-NR.sup.7R.sup.8,
C.sub.1-6alkylene-C(O)NR.sup.7R.sup.8,
C.sub.1-6alkylene-NR.sup.7C(O)R.sup.8,
C.sub.1-6alkylene-NR.sup.7C(O)NR.sup.7R.sup.8,
C.sub.1-6alkylene-S--R.sup.7, C.sub.1-6alkylene-S(O)R.sup.7,
C.sub.1-6alkylene-SO.sub.2R.sup.7,
C.sub.1-6alkylene-SO.sub.2NR.sup.7R.sup.8,
C.sub.1-6alkylene-NR.sup.7SO.sub.2R.sup.8,
C.sub.1-6alkylene-NR.sup.7SO.sub.2NR.sup.7R.sup.8,
C(O)NR.sup.7R.sup.8 and C.sub.1-6alkylene-NR.sup.7C(O)OR.sup.8,
wherein any cyclic moiety is optionally fused to a further 5- to
7-membered cyclic moiety, wherein at least one of R.sup.2, R.sup.3
and R.sup.4 is C.sub.1-6-alkylene-O--C.sub.1-6haloalkyl, and
wherein R.sup.2, R.sup.3 and R.sup.4 are optionally substituted
with one or more independently-selected R.sup.6 groups; R.sup.5 is
selected from the group consisting of H, C.sub.1-6alkyl,
C.sub.1-6haloalkyl, C.sub.2-6alkenyl, C.sub.2-6alkynyl,
C.sub.3-8cycloalkyl, C.sub.1-6alkylene-C.sub.3-8cycloalkyl, aryl,
heteroaryl, and heterocycloalkyl; R.sup.6 is selected from the
group consisting of C.sub.1-6alkyl, OH, halo,
O--(C.sub.2-3alkylene)-O, C.sub.1-6alkoxy, aryloxy,
--NH--C.sub.1-6alkyl, --N(C.sub.1-6alkyl).sub.2,
C.sub.1-6alkylene-N(C.sub.1-6alkyl).sub.2,
C.sub.1-6alkylene-NH--C.sub.1-6alkyl, cycloalkyl, heterocycloalkyl,
aryl and heteroaryl; and R.sup.7 and R.sup.8 are each independently
selected from the group consisting of H, C.sub.1-6alkyl,
C.sub.1-6haloalkyl, C.sub.2-6alkenyl, C.sub.2-6alkynyl,
C.sub.3-10cycloalkyl, C.sub.1-6alkylene-C.sub.3-10cycloalkyl,
heterocycloalkyl, aryl, C.sub.1-6alkylene-aryl,
C.sub.1-6alkylene-heterocycloalkyl, heteroaryl, and
C.sub.1-6alkylene-heteroaryl, wherein any cyclic moiety is
optionally fused to a further cyclic moiety.
2. The compound of claim 1 wherein R.sup.1 is selected from: (i)
C.sub.1-6alkyl; (ii) a substituted or unsubstituted 5- or
6-membered heteroaryl; and (iii) C.sub.3-8heterocycloalkyl, wherein
the substituents for the 5- or 6-membered heteroaryl are
independently selected from C.sub.1-4alkyl.
3. The compound of claim 1, wherein R.sup.1 is selected from: (i)
substituted or unsubstituted thiazolyl; (ii) substituted or
unsubstituted isothiazolyl; (iii) substituted or unsubstituted
oxazolyl; (iv) substituted or unsubstituted isooxazolyl; (v)
substituted or unsubstituted thiophenyl; (vi) substituted or
unsubstituted furanyl; (vii) substituted or unsubstituted
1,2,4-triazolyl; (viii) substituted or unsubstituted pyridyl; (ix)
substituted or unsubstituted pyrazinyl; (x) substituted or
unsubstituted pyrimidinyl; and (xi) substituted or unsubstituted
1,2,4-triazinyl, wherein the substituents for thiazolyl,
isothiazolyl, oxazolyl, isooxazolyl, thiophenyl, furanyl,
1,2,4-triazolyl, pyridyl, pyrazinyl, pyrimidinyl and
1,2,4-triazinyl are independently selected from C.sub.1-4alkyl.
4. The compound of claim 3, wherein R.sup.1 is selected from:
##STR00115##
5. The compound of claim 2, wherein R.sup.1 is
C.sub.3-8heterocycloalkyl and X is C.sub.1-6alkylene.
6. The compound of claim 5, wherein R.sup.1 is selected from
morpholinyl, 1,4-oxazepanyl, thiomorpholinyl, 1,4-thiazepanyl,
1,4-thiazepanyl-1-oxide, 1,4-thiazepanyl-1,1-dioxide,
1,4-thiazinanyl-1-oxide, 1,4-thiazinanyl-1,1-dioxide, aziridinyl,
azetidinyl, pyrrolidinyl, piperazinyl and 1,4-diazepanyl.
7. The compound of claim 6, wherein X is --CH.sub.2--.
8. The compound of claim 1, wherein R.sup.2 and R.sup.3 are each
independently selected from the group consisting of
C.sub.1-10alkyl, C.sub.2-10alkenyl, C.sub.2-10alkynyl,
C.sub.1-6alkylene-O--C.sub.1-6alkyl,
C.sub.1-6alkylene-O--C.sub.1-6haloalkyl,
C.sub.2-6alkenylene-O--C.sub.1-6haloalkyl and
C.sub.2-6alkynylene-O--C.sub.1-6haloalkyl, wherein at least one of
R.sup.2 and R.sup.3 is
C.sub.1-6-alkylene-O--C.sub.1-6haloalkyl.
9. The compound of claim 8, wherein R.sup.2 and R.sup.3 are each
independently selected from the group consisting of isobutyl,
--CH.sub.2--O--CH.sub.3 and --CH.sub.2--O--CHF.sub.2, wherein at
least one of R.sup.2 and R.sup.3 is --CH.sub.2--O--CHF.sub.2.
10. The compound of claim 8, wherein R.sup.4 is selected from the
group consisting of C.sub.1-10alkyl, C.sub.2-10alkenyl,
C.sub.2-10alkynyl, C.sub.1-6alkyleneC.sub.3-8cycloalkyl and
C.sub.1-6alkyleneC.sub.6-14aryl.
11. The compound of claim 1, wherein R.sup.5 is selected from the
group consisting of H and C.sub.1-6alkyl.
12. The compound of claim 1, wherein having the following relative
stereochemistry: ##STR00116##
13. The compound of claim 1, or a salt or solvate thereof, selected
from: 2-Methyl-thiazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
2-Methyl-thiazole-5-carboxylic
acid-((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarb-
amoyl]-2-methoxy-ethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
2-Methyl-thiazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-methoxyethyl)-amide;
2-Methyl-oxazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
3-Methyl-isoxazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
Thiazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxoethylcarbamoyl]-
-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)amide;
Oxazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
5-Methyl-thiophene-2-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
5-Methyl-furan-2-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
Thiophene-2-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
N-[(1S)-2-[[(1S)-2-[[(1S)-1-benzyl-2-[(2R)-2-methyloxiran-2-yl]-2-oxo-eth-
yl]amino]-1
(difluoromethoxymethyl)-2-oxo-ethyl]amino]-1-(difluoromethoxymethyl)-2-ox-
o-ethyl]-1H-1,2,4-triazole-5-carboxamide;
N-[(1S)-1-(difluoromethoxymethyl)-2-[[(1S)-1-(difluoromethoxymethyl)-2-[[-
(1S)-3-methyl-1-[(2R)-2-methyloxirane-2-carbonyl]butyl]-amino]-2-oxo-ethyl-
]amino]-2-oxo-ethyl]-2-methyl-thiazole-5-carboxamide;
Pyridine-2-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
N--((S)-1-{(S)-1-[(S)-1-Benzyl-2-((R)-2-methyl-oxiranyl)-2-oxo-ethylcarba-
moyl]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-nicotinamid-
e; Pyridine-2-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
N-[(1S)-2-[[(1S)-2-[[(1S)-1-benzyl-2-[(2R)-2-methyloxiran-2-yl]-2-oxo-eth-
yl]amino]-1-(difluoromethoxymethyl)-2-oxo-ethyl]amino]-1-(difluoromethoxym-
ethyl)-2-oxo-ethyl]-pyrimidine-2-carboxamide;
[1,2,4]Triazine-3-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarb-
amoyl]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
Pyrimidine-4-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
2-Methyl-thiazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-3-methylbutylcarbamoyl}-2-difluoromethoxyethyl)-amide;
2-Methyl-thiazole-5-carboxylic acid
((S)-2-difluoromethoxy-1-{(S)-3-methyl-1-[(S)-3-methyl-1-((R)-2-methyloxi-
ranecarbonyl)-butylcarbamoyl]-butylcarbamoyl}-ethyl)-amide,
2-Methyl-thiazole-5-carboxylic acid
((S)-1-{(S)-2-difluoromethoxy-1-[(S)-3-methyl-1-((R)-2-methyloxiranecarbo-
nyl)-butylcarbamoyl]-ethylcarbamoyl}-3-methylbutyl)-amide;
2-Methyl-thiazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-3-methylbutyl)-amide;
2-Methyl-thiazole-5-carboxylic acid
((S)-1-{(S)-2-difluoromethoxy-1-[(S)-3-methyl-1-((R)-2-methyloxiranecarbo-
nyl)-butylcarbamoyl]-ethylcarbamoyl}-2-phenylethyl)-amide;
2-Methyl-thiazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyl-oxiranyl)-2-oxo-ethylcarbamoy-
l]-2-difluoromethoxyethylcarbamoyl}-2-phenylethyl)-amide;
2S)-2-[[(2S)-3-(difluoromethoxy)-2-[(2-morpholinoacetyl)amino]propanoyl]a-
mino]-N-[(1S)-3-methyl-1-[(2R)-2-methyloxirane-2-carbonyl]butyl]-3-phenylp-
ropanamide;
(S)--N--{(S)-1-[(S)-1-Benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethyl-carbamo-
yl]-2-difluoromethoxyethyl}-3-difluoromethoxy-2-(2-morpholin-4-yl-acetylam-
ino)-propionamide;
(2S)--N-[(1S)-1-benzyl-2-[(2R)-2-methyloxiran-2-yl]-2-oxo-ethyl]-2-[[(2S)-
-3-(difluoromethoxy)-2-[(2-morpholinoacetyl)amino]propanoyl]amino]-4-methy-
lpentanamide;
(2S)-2-[[(2S)-3-(difluoromethoxy)-2-[(2-morpholinoacetyl)amino]propanoyl]-
amino]-4-methyl-N-[(1S)-3-methyl-1-[(2R)-2-methyloxirane-2-carbonyl]butyl]-
pentanamide;
(2S)--N-[(1S)-1-benzyl-2-[(2R)-2-methyloxiran-2-yl]-2-oxo-ethyl]-2-[[(2S)-
-3-(difluoromethoxy)-2-[(2-morpholinoacetyl)amino]propanoyl]amino]-3-(1-me-
thylcyclohexa-1,3,5-trien-1-yl)propanamide;
(S)-4-Methyl-2-(2-morpholin-4-yl-acetylamino)-pentanoic acid
{(S)-2-difluoromethoxy-1-[(S)-3-methyl-1-((R)-2-methyloxiranecarbonyl)-bu-
tylcarbamoyl]-ethyl}-amide;
(S)-4-Methyl-2-(2-morpholin-4-yl-acetylamino)-pentanoic acid
{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl]-2-dif-
luoromethoxyethyl}-amide;
(2S)--N-[(1S)-2-[[(1S)-1-benzyl-2-[(2R)-2-methyloxiran-2-yl]-2-oxo-ethyl]-
amino]-1(difluoromethoxymethyl)-2-oxo-ethyl]-2-[(2-morpholinoacetyl)amino]-
-3-phenylpropanamide; and
(2S)-3-(difluoromethoxy)-N-[(1S)-1-(difluoromethoxymethyl)-2-[[(1S)-3-met-
hyl-1-[(2R)-2-methyloxirane-2-carbonyl]butyl]amino]-2-oxo-ethyl]-2-[(2-mor-
pholinoacetyl)amino]-propanamide.
14. The compound of claim 13, wherein the compound is: ##STR00117##
or a salt or solvate thereof.
15. A pharmaceutical composition comprising a compound of claim 1
and a pharmaceutically acceptable carrier.
16. A method for inhibiting proteasome in a cell, comprising
administering an effective amount of a compound according to claim
1 to the cell.
17. A method of inhibiting uncontrolled and/or abnormal cellular
activities affected directly or indirectly by proteasome inhibition
in a cell, comprising administering an effective amount of a
compound according to claim 1 to the cell.
18. A method of treating a disease, disorder or condition that is
mediated by proteasome inhibition, comprising administering a
therapeutically effective amount of a compound according to claim 1
to a subject in need thereof.
19. A method of inhibiting the degradation of a protein by a
proteasome capable of degrading the protein, comprising contacting
the proteasome with an effective amount of a compound according to
claim 1.
20. A method of treating accelerated and/or enhanced proteolysis
comprising administering a therapeutically effective amount of a
compound according to claim 1 to a subject in need thereof.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation application of co-pending
U.S. patent application Ser. No. 14/420,817 filed on Feb. 10, 2015
which is a National Stage Application of International Application
No. PCT/CA2013/050620 filed on Aug. 13, 2013 which claims the
benefit of priority from U.S. Provisional Patent Application Ser.
No. 61/682,836 filed on Aug. 14, 2012, the contents of each of
which are incorporated herein by reference in their entirely.
FIELD
[0002] The present application relates to novel fluorinated
epoxyketone-based compounds, to processes for their preparation, to
compositions comprising them, and to their use in therapy. More
particularly, it relates to compounds useful in the treatment of
diseases, disorders or conditions mediated by or associated with
proteasome inhibition.
BACKGROUND
[0003] The multi-catalytic proteasome is the ubiquitous proteinase
found in cells throughout the plant and animal kingdoms that is
responsible for the ubiquitin-dependent degradation of
intracellular proteins. Thousands of copies are found in all cells,
in both the cytoplasm and the nucleus, which constitute up to 3% of
all cellular protein content. Proteasomes serve multiple
intracellular functions, including the degradation of damaged
proteins and the modulation of many regulatory proteins that affect
inflammatory processes, viral shedding, the cell cycle, growth, and
differentiation, to name but a few [Cell 1994, 79, 13-21; Nat. Rev.
Mol. Cell Biol. 2005, 6, 79-87; Semin. Oncol. 2004, 31, 3-9; Chem.
Biol. 2001, 8, 739-758].
[0004] The ubiquitin-proteasome pathway (UPP), also known as the
ubiquitin-proteasome system (UPS), regulates the degradation of
intracellular proteins with specificity as to target, time and
space. The pathway plays a central role in recognizing and
degrading misfolded and abnormal proteins in most mammalian cells
[Nature 2000, 404, 770-774]. Such a process is very important in
maintaining the biological homeostasis and regulation of different
cellular processes such as but not limited to cell differentiation,
cell cycle control, antigen processing and hormone metabolism [EMBO
J. 1998, 17, 7151-7160; Chem. Biol. 2001, 8, 739-758]. In this
pathway, the 26S proteasome is the main proteolytic component,
which is found in all eukaryotic cells and is made up of of the
cylinder-shaped multi-catalytic proteinase complex (MPC) 20S
proteasome and two regulatory particle (RP) 19S proteasomes. The
19S proteasome located at each end of the 20S proteasome is made up
of 18 subunits, and controls the recognition, unfolding, and
translocation of protein substrates into the lumen of the 20S
proteasome [Annu. Rev. Biochem. 1999, 68, 1015-1068].
[0005] X-ray crystallography of the 26S proteasome revealed that
the 20S proteasome is composed of 28 protein subunits arranged in
four stack rings, with each ring made up of seven .alpha.- and
.beta.-type subunits, following an .alpha.1-7.beta.1-7
stoichiometry [Science 1995, 268, 533-539; Nature (London) 1997,
386, 463-467]. The two outer chambers are formed by a subunits,
while the central chamber, containing the proteolytic active sites,
is made up of 1 subunits. Three of the 14 .beta. subunits are
responsible for the post-glutamyl peptide hydrolysis activity
(PGPH, attributed to .beta.1), trypsin-like activity (T-L,
.beta.2), and chymotripsin-like activity (CT-L, .beta.5),
respectively, and all these three active subunits hydrolyze the
amide bond of protein substrates with the hydrophilic
.gamma.-hydroxyl group of the N-terminal threonine
(O.gamma.-Thr1).
[0006] Rising interest in the mechanism and function of the
proteasomes and the ubiquitin system revealed that it is hard to
find any aspect of the cellular metabolic network that is not
directly or indirectly affected by the degradation system. This
includes, for example the cell cycle, the "quality control" of
newly synthesized proteins (ERAD: Endoplasmic Reticulum Associated
Protein Degradation), transcription factor regulation, gene
expression, cell differentiation and immune response as well as
pathologic processes such as cancer, neurodegenerative diseases,
lipofuscin formation, diabetes, atherosclerosis, inflammatory
processes and cataract formation in addition to the aging process
and the degradation of oxidized proteins in order to maintain cell
homeostasis. But this seems to be only a small aspect of the
general view. The various regulator proteins that are able to
change the rate or specificity of proteolysis, fitting it out for
highly specialized tasks, or the precise regulation of the
half-life of cellular proteins by ubiquitin-mediated degradation
shape the proteasome and the ubiquitin-proteasome system into a
useful part of cellular function in the three kingdoms of bacteria,
plants and animals.
[0007] Cancer is a leading cause of death worldwide. Despite
significant efforts to find new approaches for treating cancer, the
primary treatment options remain surgery, chemotherapy and
radiation therapy, either alone or in combination. Surgery and
radiation therapy, however, are generally useful only for fairly
defined types of cancer, and are of limited use for treating
patients with disseminated disease. Chemotherapy is a method that
is useful in treating patients with metastatic cancers or diffuse
cancers such as leukemias. However, although chemotherapy can
provide a therapeutic benefit, it often fails to result in cure of
the disease due to the patient's cancer cells becoming resistant to
the chemotherapeutic agent. Therefore, a need exists for additional
chemotherapeutics to treat cancer.
[0008] The concept of proteasome inhibition as a therapeutic
approach in cancer is known. The first-in-class inhibitor
bortezomib is a potent, selective, and reversible proteasome
inhibitor which targets the 26S proteasome complex and inhibits its
function. Proteasomal degradation of misfolded or damaged proteins
proceeds by recognition of poly-ubiquitinated proteins by the 19S
regulatory subunit of the 26S protease, and subsequent hydrolysis
to small polypeptides.
[0009] The successful development of bortezomib for treatment of
relapsed/refractory multiple myeloma (MM) and mantle cell lymphoma,
has shown proteasome inhibition to be a useful therapeutic strategy
[Nat. Rev. Cancer 2004, 4, 349-360; Bioorg. Med. Chem. Lett. 1998,
8, 333-338; J. Clin. Oncol. 2002, 20, 4420-4427; N. Engl. J. Med.
2003, 348, 2609-2617; N. Engl. J. Med. 2005, 352, 2487-2498; J.
Clin. Oncol. 2007, 25, 3892-3901]. Bortezomib primarily inhibits
chymotryptic, without altering tryptic or caspase-like, proteasome
activity. Bortezomib has pleiotropic effects on multiple myeloma
biology by targeting a) cell-cycle regulatory proteins; b) the
unfolded protein response (UPR) pathway via modulating the
transcriptional activity of plasma cell differentiation factor
X-box binding protein-I (XBP-I); c) p53-mediated apoptosis/MDM2; d)
DNA repair mechanisms; and e) classical stress-response pathways
via both intrinsic (caspase-9 mediated) and extrinsic (caspase-3
mediated) cell death cascades. Specifically, bortezomib activates
c-Jun N-terminal kinase (JNK), which triggers mitochondrial
apoptotic signalling: release of cytochrome-c (cyto-c) and second
mitochondrial activator of caspases (Smac) from mitochondria to
cytosol, followed by activation of caspase-9 and caspase-3.
[0010] Although bortezomib has shown clinical success, a
significant fraction of patients relapse or are refractory to
treatment [J. Clin. Oncol. 2005, 23, 676-684; J. Clin. Oncol. 2005,
23, 667-675]. Additionally, dose-limiting toxicities (DLT),
including a painful peripheral neuropathy and thrombocytopenia,
have been reported [J. Clin. Oncol. 2006, 24, 3113-3120; Blood
2005, 106, 3777-3784]. To date, it is unclear whether these
toxicities can be attributed to off-target effects because
bortezomib inhibits other enzymes such as serine proteases.
[0011] A recently reported structural analogue of the microbial
natural product epoxomicin, known as carfilzomib (also called
PR-171) was initially identified for its antitumor activity and
subsequently shown to be a potent inhibitor of the proteasome
[Cancer Res. 2007, 67, 6383-6391; Curr. Opin. Drug Discovery 2008,
11, 616-625; J. Am. Chem. Soc. 2000, 122, 1237-1238; J. Antibiot.
(Tokyo) 1992, 45, 1746-1752; Bioorg. Med. Chem. Lett. 1999, 9,
2283-2288; Cancer Res. 1999, 59, 2798-2801; Proc. Natl. Acad. Sci.
U.S.A. 1999, 96, 10403-10408]. Carfilzomib selectively inhibits the
CT-L activity of the 20S proteasome with minimal cross reactivity
to other protease classes.
[0012] Preclinical studies and phase I clinical studies
demonstrated that consecutive daily dosing schedules with
carfilzomib are both well-tolerated and promote antitumor activity
in hematologic malignancies, including patients previously treated
with bortezomib [Blood 2007, 110, 3281-3290; Br. J. Hamaetol. 2007,
136, 814-828; Blood 2007, 110, 409; Blood 2007, 110, 411].
Carfilzomib is currently being evaluated in phase I and phase II
clinical trials in multiple myeloma, non-Hodgkin's lymphoma, and
solid tumors.
[0013] Clinical responses to known proteasome inhibitor therapies
require frequent dosing (e.g., twice per week) and prolonged
treatment. For example, both bortezomib and carfilzomib are
administered intravenously (iv) on biweekly or more frequent dosing
schedules with a treatment that can extend for over 6 months.
Therefore, the development of orally bioavailable proteasome
inhibitors that would allow for dosing flexibility and improve
patient convenience is desirable.
[0014] Proteasome inhibitor-based therapeutics are useful in other
diseases beyond clinical oncology. In addition to its role in
cancer therapy, the proteasome is linked to the production of the
majority of the class I antigens [Nature 1992, 357, 375-379].
Therefore excessive inhibition of the proteasome might increase the
chance of viral infections. For example, it was reported that
replication of the HIV-1 virus could be limited by the degradative
actions of the proteasome and that the proteasome inhibitor, MG-132
or lactacystin, enhanced the ability of the virus to replicate [J.
Virol. 1998, 72, 3845-3850]. In contrast, a number of recent
publications have suggested that the ubiquitin-proteasome pathway
has a useful role in the processing of retroviral assembly,
maturation, and budding [Proc. Natl. Acad. Soc. USA 2000, 97,
13069-13074; Proc. Natl. Acad. Sci. USA 2000, 97, 13057-13062;
Proc. Natl. Acad. Sci. USA 2000, 97, 13063-13068]. Proteasome
inhibition also interferes with gag polyprotein processing,
release, and maturation of HIV-1 and HIV-2, and ubiquitination is
required for retroviral release. Hence, proteasome inhibitors can
be useful for the treatment of HIV and other viral infections.
[0015] Proteasome inhibition also has clinical potential for
treatment of inflammatory and autoimmune diseases through multiple
pathways, including MHC-mediated antigen presentation, cytokine and
cell cycle regulation, and apoptosis [J. Rheumatol. 2005, 32,
1192-119]. In inflammatory arthritis, it was shown that NF-.kappa.B
regulates multiple critical cytokines involved in the pathogenesis
of rheumatoid arthritis (RA) [Arthritis Rheum. 2004, 50, 2381-2386;
Arthritis Rheum. 2004, 50, 3541-3548]. In the
peptoglycan/polysaccharide-induced inflammatory arthritis model, a
proteasome inhibitor improved the arthritis score by suppressing
the activation of NF-.kappa.B, reducing the expression of cell
adhesion molecules and IL-6. In addition, proteasome inhibition may
regulate the development of inflammatory arthritis by controlling
angiogenesis [J. Mol. Med. 2003, 81, 235-245].
[0016] Psoriasis is one of the prototypical T cell-mediated
diseases, and its development is related to the activation of
NF-.kappa.B. Administration of a proteasome inhibitor has been
reported to reduce the size of psoriatic lesions in human skin
explants grafted onto mice. The treatment also resulted in reduced
super antigen-mediated T-cell activation, attenuated cell adhesion
molecule expression and decreased expression of T-cell activation
markers that were significantly elevated during the disease process
[J. Clin. Invest. 2002, 109, 671-679].
[0017] In addition, other studies showed oral proteasome inhibition
by bortezomib significantly limited overall inflammation, reduced
the activation of NF-.kappa.B, lowered cell adhesion molecule
expression, inhibited nitric oxide synthase activity, attenuated
the circulating levels of IL-6, reduced the arthritic index and
swelling observed in the joints of the animals, and improved the
histologic appearance of the joints compared with vehicle-treated
animals [Carcinogenesis 2000, 2, 505-515].
[0018] A link between proteasome inhibition, allergy and asthma has
also been shown. Abnormal activation of type 2 helper T cells (Th2)
results in asthmatic and allergic symptoms [Nat. Immunol. 2002, 3,
715-720]. E3 ubiquitin ligase Itch plays a useful role in
maintaining immune tolerance mediated through Th2 cells both in
vitro and in vivo. Itch deficient mice failed to block the
development of airway inflammation in an allergic model [J. Clin.
Invest. 2006, 116, 1117-1126]. Consistent with these findings,
useful therapeutic effects were observed in a rodent model of
allergen-induced asthma [J. Allergy Clin. Immunol. 1999, 104,
294-300].
[0019] Other inflammatory and autoimmune diseases have been linked
to the ubiquitin-proteasome system (UPS), such as seronegative
spondyloarthropathies (SpA) which are a group of diseases
characterized by, but not limited to, axial joint inflammation.
Ankylosing spondylitis (AS) is the prototypical SpA. Most patients
with AS carry the MHC class I HLA-B27 gene, and therefore much
research effort has been directed at understanding the role of this
gene in the disease pathogenesis. There has also been interest
focused on determining the origin and nature of the peptides being
presented by HLA-B27 and the cell surface expression of misfolded
HLA-B27, two areas in which the UPS is known to play a role.
[0020] The UPS is involved in the regulation or induction of
apoptosis. Apoptosis has been implicated in both experimental
models and clinical systemic lupus erythematosus (SLE). In mature,
activated lymphocytes, the proteasome inhibitor lactacystin induces
DNA fragmentation and apoptosis in a dose-dependent fashion,
indicating that proteasome suppresses apoptosis in these cells.
Altered clearance of auto antigens is thought to allow for
targeting by the immune system and the development of autoimmunity.
The involvement of UPS in regulating the levels of Ku70 and other
autoantigens has been reported [J. Biol. Chem. 1998, 273,
31068-31074; J. Cell. Sci. 1994, 107 (Pt 11), 3223-3233; Exp. Cell.
Res. 2006, 312, 488-499].
[0021] Proteasome inhibition has also been linked to heart disease.
Evidence continues to emerge to support a hypothesis that
proteasome functional insufficiency represents a common
pathological phenomenon in a large subset of heart disease,
compromises protein quality control in heart muscle cells, and
thereby acts as a major pathogenic factor promoting the progression
of the subset of heart disease to congestive heart failure. This
front is represented by the studies on the UPS in cardiac
proteinopathy, which have taken advantage of a transgenic mouse
model expressing a fluorescence reporter for UPS proteolytic
function.
[0022] In addition, pharmacological inhibition of the proteasome
has been explored experimentally as a potential therapeutic
strategy to intervene on some forms of heart disease, such as
pressure-overload cardiac hypertrophy, viral myocarditis, and
myocardial ischemic injury [Biochimica et Biophysica Acta--Gene
Regulatory Mechanisms, 1799(9), 2010, 597-668]. Furthermore,
initial reports on the effects of proteasome inhibitors in
cardiovascular diseases, indicate that proteasome inhibition might
be a useful therapeutic strategy for the reduction of the
proliferative phenomena of the progression stage of atherogenesis
[Cardiovasc. Res. 2004, 61, 11-21]. Recent data on the improvement
of endothelium-dependent vasorelaxation in vitro, correlating with
an increase in endothelial nitric oxide synthase (eNOS) expression,
suggest a therapeutic potential of proteasome inhibition in the
early stages of atherosclerosis [FASEB 2004, 18, 272-279].
[0023] Proteasome inhibitors have been shown to exert a substantial
anti-inflammatory effect, which was attributed to a reduction in
the activity of the factor NF-.kappa.B [Cardiovasc. Res. 2004, 61,
11-21]. As the pathogenesis of cardiovascular events in diabetic
patients involves inflammation, the use of proteasome inhibitors
may be a useful therapy. In addition to epidemiological evidence
for the role of inflammation in diabetes-associated cardiovascular
events, clinical studies of patients on cardio-protective drug
regimens have revealed that many of the pharmacotherapies mediate
their benefits, at least in part, through anti-inflammatory
activities. This is the case for one class of drugs that improves
adipose tissue physiology and insulin sensitivity, the peroxisome
proliferator-activated receptor-.gamma. (PPAR.gamma.) agonists
[Arterioscler. Thromb. Vasc. Biol. 2002, 22, 717-726]. For example,
the PPAR.gamma. agonist rosiglitazone, reducing inflammation, may
prevent plaque progression to an unstable phenotype in diabetic
patients with asymptomatic carotid stenosis, enlisted to undergo
carotid endarterectomy for extracranial high-grade (>70%)
internal carotid artery stenosis [Diabetes 2006, 55, 622-632].
[0024] The anti-inflammatory effects of glitazones are felt to be
mediated partly by their beneficial effects on glycemia, but there
is also evidence that glitazones may directly modulate inflammation
via transcription factors such as NF-.kappa.B [Arterioscler.
Thromb. Vasc. Biol. 2002, 22, 717-726]. In line with this, recent
data have shown an inhibitory effect of rosiglitazone on
ubiquitin-proteasome activity in diabetic lesions [Diabetes 2006,
55, 622-632]. At the same level of blood glucose levels, diabetic
patients treated with rosiglitazone had the lowest level of
ubiquitin and proteasome 20S activity, plaque inflammatory cells,
cytokines, oxidative stress and MMP-9 associated with the highest
content of plaque interstitial collagen. Patients assigned to
rosiglitazone had lesser plaque progression to an unstable
phenotype compared with patients assigned to placebo.
[0025] For aspirin and statins, two of the most successful drugs in
treatment of cardiovascular diseases, a proteasome inhibitory
effect has been described [Mol. Pharmacol. 2002, 62,
1515-1521].
[0026] Drugs that modulate the proteasomal degradation of proteins
could be useful agents for the treatment of insulin-resistant and
type-2 diabetes, and pharmacological therapies targeting UPS
activity may be useful in the treatment of vascular biology
disorders associated with diabetes [Cardiovascular Diabetology
2007, 6:35, 1-9].
[0027] The ubiquitin-proteasome system is also believed to degrade
the major contractile skeletal muscle proteins and plays a major
role in muscle wasting. Different and multiple events in the
ubiquitination, deubiquitination and proteolytic machineries are
responsible for the activation of the system and subsequent muscle
wasting. However, other proteolytic enzymes act upstream (possibly
m-calpain, cathepsin L, and/or caspase-3) and downstream
(tri-peptidyl-peptidase II and amino-peptidases) of the UPS, for
the complete breakdown of the myofibrillar proteins into free amino
acids. Recent studies have identified a few proteins that seem
necessary for muscle wasting i.e. the MAFbx (muscle atrophy F-box
protein, also called atrogin-1) and MuRF-1 (muscle-specific RING
ubiquitin-protein ligases) proteins. The characterization of their
signaling pathways is leading to new pharmacological approaches
that can be useful to block or partially prevent muscle wasting in
human patients [Essays Biochem. 2005, 41, 173-86].
[0028] The UPS has also been linked to the development of human
obesity. For example, it was shown that there is a possible
correlation between plasma ubiquitin, 26S proteasome levels, and
obesity. The body mass index (BMI), plasma ubiquitin levels, and
26S proteasome activity levels were determined and statistically
analyzed. Comparison of the immunoglobulin among the underweight,
normal weight, and overweight groups demonstrated that plasma
ubiquitin is significantly decreased in obese individuals versus
normal controls, and plasma ubiquitin levels were found to be
inversely correlated with the BMI. In addition, there was an
inverse relationship between 20S proteasome levels in red blood
cells and BMI, whereas 26S proteasome activity was found to be
dependent quantitatively to S5a in erythrocytes. Furthermore,
immunoglobulin is significantly decreased in overweight individuals
versus normal controls [Metabolism 2009, 58(11), 1643-8].
[0029] A wide variety of preclinical and early clinical studies
have been performed to test the potential usefulness of proteasome
inhibitors for the treatment of neurodegenerative disorders,
including Alzheimer's (AD) and Parkinson's (PD) diseases. These CNS
disorders are characterized by a selective loss of neurons in
specific, but different, regions of the brain, and the result is
often a disruption to motor, sensory or cognitive systems,
resulting in severe disability of the patient. The pathological
characteristic of many neurodegenerative diseases is the presence
of distinctive ubiquitin-positive, intra- or extracellular
inclusion bodies in affected regions of the brain. In general,
these inclusions are made up of insoluble, unfolded, ubiquitylated
polypeptides that fail to be targeted and degraded by the 26S
proteasome [J. Pathol. 1988, 155, 9-15; Neuron 2001, 29, 15-32].
Their apparent stability may, in part, be due to decreased levels
of 26S proteasomal activity that is associated with increasing age
[Ann. N. Y. Acad. Sci. 2001, 928, 54-64].
[0030] Proteins associated with the UPS are now known to play
either a direct or indirect role in familial forms of
neurodegenerative disease and, in particular, PD. UPS-mediated
post-translational modification and degradation of proteins is
useful for most cellular processes such as cell cycling, DNA
repair, cell signaling, gene transcription and apoptosis.
Historically, it was recognized that the UPS is the major route by
which proteins are selected for temporal and spatial degradation in
eukaryotic organisms [Cell 2004, 116, 181-190; Nat. Rev. Mol. Cell
Biol. 2003, 4, 192-201]. The key constituents of the inclusions
associated with neurodegenerative disorders are mis-folded
proteins. The major causes of protein mis-folding and subsequent
loss of function are mis-sense mutations, modifications or
posttranslational damage of proteins, or expansion of amino acid
repeats as is observed in polyglutamine (polyQ) disorders such as
Huntington's disease (HD).
[0031] Of all the neurodegenerative diseases, PD is most closely
associated with aberrant protein processing via the UPS. Indeed, of
the known proteins associated with hereditary forms of PD, Parkin
and UCH-L1 are components of the UPS, whereas modified and/or
mutant .alpha.-Synuclein and DJ-1 are degraded by the system
[Nature 1998, 392, 605-608; Nature 1998, 395, 451-452; J. Biol.
Chem. 2003, 278, 36588-36595].
[0032] A wide variety of preclinical and early clinical studies
have been performed to test the potential usefulness of proteasome
inhibitors for the treatment of Alzheimer's disease [J. Neurochem.
1999, 72, 255-261], amyotrophic lateral sclerosis [J. Neurol. Sci.
1996, 139, 15-20], autoimmune thyroid disease [Tissue Antigens.
1997, 50, 153-163], cachexia [N. Engl. J. Med. 1996, 335,
1897-1905; Am. J. Physiol. 1999, 277, 332-341], Crohn's disease [J.
Pharmacol. Exp. Ther. 1997, 282, 1615-1622], Hepatitis B [Oncogene,
1998, 16, 2051-2063], inflammatory bowel disease [Inflamm. Bowel
Dis. 1996, 2, 133-147], sepsis [Ann. Surg. 1997, 225, 307-316],
systemic lupus erythematosus [J. Exp. Med. 1996, 10, 1313-1318],
and transplantation rejection and related immunology [Drug Discov.
Today 1999, 4, 63-70; Transplantation 2001, 72, 196-202].
[0033] The ubiquitin-proteasome system is also believed to play
roles in the pathogenesis of eye diseases. Accumulation of the
cytotoxic abnormal proteins in eye tissues is etiologically
associated with many age-related eye diseases such as retina
degeneration, cataract, and certain types of glaucoma. Age- or
stress-induced impairment or overburdening of the UPP appears to
contribute to the accumulation of abnormal proteins in eye tissues.
Cell cycle and signal transduction are regulated by the conditional
UPP-dependent degradation of the regulators of these processes.
Impairment or overburdening of the UPP could also result in
dysregulation of cell cycle control and signal transduction. The
consequences of the improper cell cycle and signal transduction
include defects in ocular development, wound healing, angiogenesis,
or inflammatory responses. Methods that enhance or preserve UPP
function or reduce its burden may be useful strategies for
preventing age-related eye diseases [Pro. Mol. Biol. & Trans.
Sc., Vol. 109, 2012, 347-396].
[0034] The search for subunit selective inhibitors is predominantly
conducted by either screening of natural products [Bioorg. Med.
Chem. Lett. 1999, 9, 3335-3340], rational design [Chem. Biol. 2009,
16, 1278-1289], or compound library building [Proc. Natl. Acad.
Sci. USA 2001, 98, 2967-2972; Org. Biomol. Chem. 2007, 5,
1416-1426]. It was noted that in these studies the effect of
fluorine functionality in proteasome inhibitors is relatively
uncharted [Bioorg. Med. Chem. Lett. 2009, 19, 83-86].
[0035] The epoxomicin analog PR-047 was recently reported to be an
orally-bioavailable candidate that displayed moderate to poor
metabolic properties [J. Med. Chem. 2009, 52, 3028-3038]. While not
wishing to be limited by theory, this poor metabolic property is
thought to be due to the methoxy groups in the serine (OMe)
side-chains undergoing demethylation to the O-desmethyl metabolite.
A need therefore exists to find a route to block this demethylation
pathway to give compounds having useful clinical profile.
SUMMARY
[0036] A novel class of halogenated epoxyketone-based proteasome
inhibitors of Formula I has been prepared and found to be useful in
the treatment of cancers and other proteasome mediated or
associateddisorders.
[0037] Accordingly, the present application includes a compound of
Formula I or a pharmaceutically acceptable salt, solvate or prodrug
thereof:
##STR00002##
wherein:
[0038] R.sup.1 is selected from the group consisting of
C.sub.1-10alkyl, C.sub.2-10alkenyl, C.sub.2-10alkynyl,
C.sub.1-10haloalkyl, C.sub.1-10cyanoalkyl, C.sub.1-10alkoxy,
C.sub.2-10alkenyloxy, C.sub.2-10alkynyloxy, C.sub.3-10cycloalkyl,
heterocycloalkyl, aryl, heteroaryl,
C.sub.1-6alkylene-O--C.sub.1-6alkyl,
C.sub.1-6alkylene-O--C.sub.1-6haloalkyl,
C.sub.2-6alkenylene-O--C.sub.1-6haloalkyl,
C.sub.2-6alkynylene-O--C.sub.1-6haloalkyl,
C.sub.1-6alkylene-C.sub.3-8cycloalkyl,
C.sub.1-6alkylene-heterocycloalkyl, C.sub.1-6alkylene-aryl,
C.sub.1-6alkylene-heteroaryl, C(O)R.sup.7, OC(O)R.sup.7,
C(O)OR.sup.7, C.sub.1-6alkylene-O--R.sup.7,
C.sub.1-6alkylene-C(O)R.sup.7, C.sub.1-6alkylene-O--C(O)R.sup.7,
C.sub.1-6alkylene-C(O)OR.sup.7, C.sub.1-6alkylene-O--C(O)OR.sup.7,
C.sub.1-6alkylene-NR.sup.7R.sup.8,
C.sub.1-6alkylene-C(O)NR.sup.7R.sup.8,
C.sub.1-6alkylene-NR.sup.7C(O)R.sup.8,
C.sub.1-6alkylene-NR.sup.7C(O)NR.sup.7R.sup.8,
C.sub.1-6alkylene-S--R.sup.7, C.sub.1-6alkylene-S(O)R.sup.7,
C.sub.1-6alkylene-SO.sub.2R.sup.7,
C.sub.1-6alkylene-SO.sub.2NR.sup.7R.sup.8,
C.sub.1-6alkylene-NR.sup.7SO.sub.2R.sup.8,
C.sub.1-6alkylene-NR.sup.7SO.sub.2NR.sup.7R.sup.8,
C(O)NR.sup.7R.sup.8 and C.sub.1-6alkylene-NR.sup.7C(O)OR.sup.8,
wherein any cyclic moiety is optionally substituted with
C.sub.1-4alkyl and/or is optionally fused to a further cyclic
moiety;
[0039] X is absent or is selected from the group consisting of O,
NH, NC.sub.1-6alkyl, S, S(O), SO.sub.2, C(O), C.sub.1-6alkylene,
C.sub.2-6alkenylene, C.sub.2-6alkynylene, C.sub.1-6haloalkylene,
C.sub.3-8cycloalkylene, heterocycloalkylene, arylene and
heteroarylene, or X is a combination of two or three of O, NH,
NC.sub.1-6alkyl, S, S(O), SO.sub.2, C.sub.1-6alkylene,
C.sub.2-6alkenylene, C.sub.2-6alkynylene, C.sub.1-6haloalkylene,
C.sub.3-8cycloalkylene, heterocycloalkylene, arylene or
heteroarylene, bonded together in a linear fashion, provided that
two or three of O, NH, NC.sub.1-6alkyl, S, S(O) and SO.sub.2 and
not bonded directly to each other;
[0040] R.sup.2, R.sup.3 and R.sup.4 are each independently selected
from the group consisting of C.sub.1-10alkyl, C.sub.2-10alkenyl,
C.sub.2-10alkynyl, C.sub.1-10haloalkyl, C.sub.1-10cyanoalkyl,
C.sub.1-10alkoxy, C.sub.2-10alkenyloxy, C.sub.2-10alkynyloxy,
C.sub.3-10cycloalkyl, heterocycloalkyl, aryl, heteroaryl,
C.sub.1-6alkylene-O--C.sub.1-6alkyl,
C.sub.1-6alkylene-O--C.sub.1-6haloalkyl,
C.sub.2-6alkenylene-O--C.sub.1-6haloalkyl,
C.sub.2-6alkynylene-O--C.sub.1-6haloalkyl,
C.sub.1-6alkylene-C.sub.3-8cycloalkyl,
C.sub.1-6alkylene-heterocycloalkyl, C.sub.1-6alkylene-aryl,
C.sub.1-6alkylene-heteroaryl, C(O)R.sup.7, OC(O)R.sup.7,
C(O)OR.sup.7, C.sub.1-6alkylene-O--R.sup.7,
C.sub.1-6alkylene-C(O)R.sup.7, C.sub.1-6alkylene-O--C(O)R.sup.7,
C.sub.1-6alkylene-C(O)OR.sup.7, C.sub.1-6alkylene-O--C(O)OR.sup.7,
C.sub.1-6alkylene-NR.sup.7R.sup.8,
C.sub.1-6alkylene-C(O)NR.sup.7R.sup.8,
C.sub.1-6alkylene-NR.sup.7C(O)R.sup.8,
C.sub.1-6alkylene-NR.sup.7C(O)NR.sup.7R.sup.8,
C.sub.1-6alkylene-S--R.sup.7, C.sub.1-6alkylene-S(O)R.sup.7,
C.sub.1-6alkylene-SO.sub.2R.sup.7,
C.sub.1-6alkylene-SO.sub.2NR.sup.7R.sup.8,
C.sub.1-6alkylene-NR.sup.7SO.sub.2R.sup.8,
C.sub.1-6alkylene-NR.sup.7SO.sub.2NR.sup.7R.sup.8,
C(O)NR.sup.7R.sup.8 and C.sub.1-6alkylene-NR.sup.7C(O)OR.sup.8,
wherein any cyclic moiety is optionally fused to a further 5- to
7-membered cyclic moiety, wherein at least one of R.sup.2, R.sup.3
and R.sup.4 is C.sub.1-6-alkylene-O--C.sub.1-6haloalkyl, and
wherein R.sup.2, R.sup.3 and R.sup.4 are optionally substituted
with one or more independently-selected R.sup.6 groups;
[0041] R.sup.5 is selected from the group consisting of H,
C.sub.1-6alkyl, C.sub.1-6haloalkyl, C.sub.2-6alkenyl,
C.sub.2-6alkynyl, C.sub.3-8cycloalkyl,
C.sub.1-6alkylene-C.sub.3-8cycloalkyl, aryl, heteroaryl, and
heterocycloalkyl;
[0042] R.sup.6 is selected from the group consisting of
C.sub.1-6alkyl, OH, halo, O--(C.sub.2-3alkylene)-O,
C.sub.1-6alkoxy, aryloxy, --NH--C.sub.1-6alkyl,
--N(C.sub.1-6alkyl).sub.2,
C.sub.1-6alkylene-N(C.sub.1-6alkyl).sub.2,
C.sub.1-6alkylene-NH--C.sub.1-6alkyl, cycloalkyl, heterocycloalkyl,
aryl and heteroaryl; and
[0043] R.sup.7 and R.sup.8 are each independently selected from the
group consisting of H, C.sub.1-6alkyl, C.sub.1-6haloalkyl,
C.sub.2-6alkenyl, C.sub.2-6alkynyl, C.sub.3-10cycloalkyl,
C.sub.1-6alkylene-C.sub.3-10cycloalkyl, heterocycloalkyl, aryl,
C.sub.1-6alkylene-aryl, C.sub.1-6alkylene-heterocycloalkyl,
heteroaryl, and C.sub.1-6alkylene-heteroaryl, wherein any cyclic
moiety is optionally fused to a further cyclic moiety.
[0044] The present application also includes a composition
comprising one or more compounds of the application and a carrier.
In an embodiment, the composition is a pharmaceutical composition
comprising one or more compounds of the application and a
pharmaceutically acceptable carrier.
[0045] The compounds of the application have been shown to
inhibitors of proteasome activity. Therefore the compounds of the
application are useful for treating diseases, disorders or
conditions mediated by or associated with proteasome inhibition.
Accordingly, the present application also includes a method of
treating a disease, disorder or condition mediated by proteasome
inhibition, comprising administering a therapeutically effective
amount of one or more compounds of the application to a subject in
need thereof.
[0046] In a further embodiment, the compounds of the application
are used as medicaments. Accordingly, the application also includes
a compound of the application for use as a medicament.
[0047] The present application also includes a use of one or more
compounds of the application for treatment of a disease, disorder
or condition mediated by proteasome inhibition as well as a use of
one or more compounds of the application for the preparation of a
medicament for treatment of a disease, disorder or condition
mediated by proteasome inhibition. The application further includes
one or more compounds of the application for use in treating a
disease, disorder or condition mediated by proteasome
inhibition.
[0048] In an embodiment, the disease, disorder or condition
mediated by proteasome inhibition is a neoplastic disorder. In an
embodiment, the treatment is in an amount effective to ameliorate
at least one symptom of the neoplastic disorder, for example,
reduced cell proliferation or reduced tumor mass in a subject in
need of such treatment.
[0049] In an embodiment, the disease, disorder or condition
mediated by proteasome inhibition is cancer.
[0050] In an embodiment, the disease, disorder or condition
mediated by proteasome inhibition is a disease, disorder or
condition associated with an uncontrolled and/or abnormal cellular
activity affected directly or indirectly by proteasome inhibition.
In another embodiment, the uncontrolled and/or abnormal cellular
activity that is affected directly or indirectly by proteasome
inhibition is proliferative activity in a cell.
[0051] The application also includes a method of inhibiting
proliferative activity in a cell, comprising administering an
effective amount of one or more compounds of the application to the
cell.
[0052] In a further embodiment the disease, disorder or condition
mediated by proteasome inhibition is cancer and the one or more
compounds of the application are administered in combination with
one or more additional cancer treatments. In another embodiment,
the additional cancer treatment is selected from radiotherapy,
chemotherapy, targeted therapies such as antibody therapies and
small molecule therapies such as tyrosine-kinase inhibitors,
immunotherapy, hormonal therapy and anti-angiogenic therapies.
[0053] In another embodiment, the disease, disorder or condition
mediated by proteasome inhibition is selected from by proteasome
inhibition is selected from a viral infection, an inflammatory
disease, an autoimmune disease, heart disease, an age-related eye
disease and a neurodegenerative disease.
[0054] The application additionally provides a process for the
preparation of compounds of Formula I. General and specific
processes are discussed in more detail and set forth in the
Examples below.
[0055] In an embodiment of the present application, the compounds
of Formula I comprise at least one fluorine atom. Factors to be
considered when synthesising fluorine-containing compounds include
(a) the relatively small size of the fluorine atom (van der Waals
radius of 1.47 .ANG.), comparable to hydrogen (van der Waals radius
of 1.20 .ANG.), (b) the highly electron-withdrawing nature of
fluorine, (c) the greater stability of the C--F bond compared to
the C--H bond and (d) the greater lipophilicity of fluorine
compared to hydrogen. The introduction of a fluorine atom into a
molecule can alter the physicochemical properties of the compound
due to its electronegativity.
[0056] The introduction of a halogen atom into a molecule also
provides the opportunity for the use of the molecule in
radiolabeling applications. For example, .sup.18F is used as a
radiolabel tracer in the sensitive technique of Positron Emission
Tomography (PET). Accordingly the present application also includes
methods of using the compounds of Formula I for diagnostic and
imaging purposes.
[0057] Other features and advantages of the present application
will become apparent from the following detailed description. It
should be understood, however, that the detailed description and
the specific examples while indicating embodiments of the
application are given by way of illustration only, since various
changes and modifications within the spirit and scope of the
application will become apparent to those skilled in the art from
this detailed description.
BRIEF DESCRIPTION OF THE DRAWINGS
[0058] The present application will now be described in greater
detail with reference to the drawings, in which:
[0059] FIG. 1 shows the effect of a representative compound of the
application (the compound of Example 1) on the enzymatic activity
of purified proteasomes, compared to bortezomib. Purified beta
proteasome subunits from the archaebacteria Thermoplasma
acidophilum (A) or whole cell lysates from the human myeloma cell
line LP1 (B).
[0060] FIG. 2 shows the compound of Example 1 synergized with
standard antimyeloma therapeutics when combined in a fixed ratio
for 72 hours at low fractional effect levels. Synergy was assessed
using the MTS assay and combination index (CI) analysis, where a
CI<1 indicates synergy between two drugs, a CI=1 indicates
additivity, and a CI>1 indicates antagonism.
[0061] FIG. 3 shows the effects of the compound of Example 1 on the
cell viability of samples from patients with multiple myeloma (A,C)
or from the peripheral blood of a patient with plasma cell leukemia
(B).
DETAILED DESCRIPTION
I. Definitions
[0062] Unless otherwise indicated, the definitions and embodiments
described in this and other sections are intended to be applicable
to all embodiments and aspects of the application herein described
for which they are suitable as would be understood by a person
skilled in the art. Unless otherwise specified within this
application or unless a person skilled in the art would understand
otherwise, the nomenclature used in this application generally
follows the examples and rules stated in "Nomenclature of Organic
Chemistry" (Pergamon Press, 1979), Sections A, B, C, D, E, F, and
H. Optionally, a name of a compound may be generated using a
chemical naming program: ACD/ChemSketch, Version 5.09/September
2001, Advanced Chemistry Development, Inc., Toronto, Canada.
[0063] The term "compound of the application" or "compound of the
present application" and the like as used herein refers to a
compound of Formula I, or a pharmaceutically acceptable salt,
solvate and/or prodrug thereof.
[0064] As used in the present application, the singular forms "a",
"an" and "the" include plural references unless the content clearly
dictates otherwise. For example, an embodiment including "a
compound" should be understood to present certain aspects with one
compound, or two or more additional compounds.
[0065] In embodiments comprising an "additional" or "second"
component, such as an additional or second compound, the second
component as used herein is chemically different from the other
components or first component. A "third" component is different
from the other, first, and second components, and further
enumerated or "additional" components are similarly different.
[0066] In understanding the scope of the present application, the
term "comprising" and its derivatives, as used herein, are intended
to be open ended terms that specify the presence of the stated
features, elements, components, groups, integers, and/or steps, but
do not exclude the presence of other unstated features, elements,
components, groups, integers and/or steps. The foregoing also
applies to words having similar meanings such as the terms,
"including", "having" and their derivatives. The term "consisting"
and its derivatives, as used herein, are intended to be closed
terms that specify the presence of the stated features, elements,
components, groups, integers, and/or steps, but exclude the
presence of other unstated features, elements, components, groups,
integers and/or steps. The term "consisting essentially of", as
used herein, is intended to specify the presence of the stated
features, elements, components, groups, integers, and/or steps as
well as those that do not materially affect the basic and novel
characteristic(s) of features, elements, components, groups,
integers, and/or steps.
[0067] The term "suitable" as used herein means that the selection
of the particular compound or conditions would depend on the
specific synthetic manipulation to be performed, and the identity
of the species to be transformed, but the selection would be well
within the skill of a person trained in the art. All method steps
described herein are to be conducted under conditions sufficient to
provide the desired product. A person skilled in the art would
understand that all reaction conditions, including, for example,
reaction solvent, reaction time, reaction temperature, reaction
pressure, reactant ratio and whether or not the reaction should be
performed under an anhydrous or inert atmosphere, can be varied to
optimize the yield of the desired product and it is within their
skill to do so.
[0068] In embodiments of the present application, the compounds
described herein have at least one asymmetric center. Where
compounds possess more than one asymmetric center, they may exist
as diastereomers. It is to be understood that all such isomers and
mixtures thereof in any proportion are encompassed within the scope
of the present application. It is to be further understood that
while the stereochemistry of the compounds may be as shown in any
given compound listed herein, such compounds may also contain
certain amounts (for example, less than 20%, suitably less than
10%, more suitably less than 5%) of compounds of the present
application having alternate stereochemistry. It is intended that
any optical isomers, as separated, pure or partially purified
optical isomers or racemic mixtures thereof are included within the
scope of the present application.
[0069] In embodiments of the present application, the compounds
described herein having a double bond can exist as geometric
isomers, for example cis or trans isomers. It is to be understood
that all such geometric isomers and mixtures thereof in any
proportion are encompassed within the scope of the present
application. It is to be further understood that while the
stereochemistry of these compounds may be as shown in any given
compound listed herein, such compounds may also contain certain
amounts (for example, less than 20%, suitably less than 10%, more
suitably less than 5%) of compounds of the present application
having alternate stereochemistry.
[0070] The compounds of the present application can also exist in
different tautomeric forms and it is intended that any tautomeric
forms which the compounds form, are included within the scope of
the present application.
[0071] The compounds of the present application may further exist
in varying polymorphic forms and it is contemplated that any
polymorphs which form, are included within the scope of the present
application.
[0072] Terms of degree such as "substantially", "about" and
"approximately" as used herein mean a reasonable amount of
deviation of the modified term such that the end result is not
significantly changed. These terms of degree should be construed as
including a deviation of at least .+-.5% of the modified term if
this deviation would not negate the meaning of the word it modifies
or unless the context suggests otherwise to a person skilled in the
art.
[0073] The expression "proceed to a sufficient extent" as used
herein with reference to the reactions or method steps disclosed
herein means that the reactions or method steps proceed to an
extent that conversion of the starting material or substrate to
product is maximized. Conversion may be maximized when greater than
about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75,
80, 85, 90, 95 or 100% of the starting material or substrate is
converted to product.
[0074] The term "seven-membered" or "7-membered" as used herein as
a prefix refers to a group having a ring that contains seven ring
atoms.
[0075] The term "six-membered" or "6-membered" as used herein as a
prefix refers to a group having a ring that contains six ring
atoms.
[0076] The term "five-membered" or "5-membered" as used herein as a
prefix refers to a group having a ring that contains five ring
atoms.
[0077] The term "hydrocarbon" as used herein, whether it is used
alone or as part of another group, refers to any structure
comprising only carbon and hydrogen atoms up to 14 carbon
atoms.
[0078] The term "hydrocarbon radical" or "hydrocarbyl" as used
herein, whether it is used alone or as part of another group,
refers to any structure derived as a result of removing a hydrogen
atom from a hydrocarbon.
[0079] The term "hydrocarbylene" as used herein, whether it is used
alone or as part of another group, refers to any structure derived
as a result of removing a hydrogen atom from two ends of a
hydrocarbon.
[0080] The term "alkyl" as used herein, whether it is used alone or
as part of another group, means straight or branched chain,
saturated hydrocarbyl groups. For example, the term C.sub.1-10alkyl
means an alkyl group having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon
atoms.
[0081] The term "alkylene" as used herein means straight or
branched chain, saturated hydrocarbyl group, that is a saturated
carbon chain that contains substituents on two of its ends. For
example, the term C.sub.1-10alkylene means an alkylene group having
1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms.
[0082] The term "alkenyl" as used herein, whether it is used alone
or as part of another group, means straight or branched chain,
unsaturated alkenyl groups. For example, the term C.sub.2-10alkenyl
means an alkenyl group having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon
atoms and at least one double bond, for example 1-3, 1-2 or 1
double bond.
[0083] The term "alkenylene" as used herein means straight or
branched chain, unsaturated alkenylene group, that is an
unsaturated carbon chain that contains substituents on two of its
ends. For example, the term C.sub.2-6alkenylene means an alkenylene
group having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms and at least
1, for example 1-3, 1-2 or 1 double bond.
[0084] The term "alkynyl" as used herein, whether it is used alone
or as part of another group, means straight or branched chain
unsaturated alkynyl groups. The term C.sub.2-6alkynyl means an
alkynyl group having 2, 3, 4, 5 or 6 carbon atoms and at least one
triple bond, for example 1-3, 1-2 or 1 triple bond.
[0085] The term "alkynylene" as used herein means straight or
branched chain, unsaturated alkynylene group, that is an
unsaturated carbon chain that contains substituents on two of its
ends. The term C.sub.2-6alkynylene means an alkynylene group having
2, 3, 4, 5 or 6 carbon atoms and at least 1, for example 1-3, 1-2
or 1 triple bond.
[0086] The term "haloalkyl" as used herein refers to an alkyl group
wherein one or more, including all of the hydrogen atoms are
replaced by a halogen atom. In an embodiment, the halogen is
fluorine, in which case the haloalkyl is referred to herein as a
"fluoroalkyl" group. In another embodiment, the haloalkyl comprises
at least one --CHF.sub.2 group.
[0087] The term "haloalkylene" as used herein refers to an alkylene
group wherein one or more, including all of the hydrogen atoms are
replaced by a halogen atom. In an embodiment, the halogen is
fluorine, in which case the haloalkylene is referred to herein as a
"fluoroalkylene" group. In another embodiment, the haloalkylene
comprises a branched fluoroalkylene having at least one --CHF.sub.2
group.
[0088] The term "cyanoalkyl" as used herein refers to an alkyl
group that is substituted by at least one cyano group. For example,
the term C.sub.1-10cyanoalkyl means an alkyl group having 1, 2, 3,
4, 5, 6, 7, 8, 9 or 10 carbon atoms and at least one cyano group
attached thereto.
[0089] The term "alkoxy" as used herein, whether it is used alone
or as part of another group, refers to the group "alkyl-O--". For
example, the term C.sub.1-10alkoxy means an alkyl group having 1,
2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms bonded to the oxygen atom
of the alkoxy group. Exemplary alkoxy groups include without
limitation methoxy, ethoxy, propoxy, isopropoxy, butoxy, t-butoxy
and isobutoxy.
[0090] The term "alkenyloxy" as used herein, whether it is used
alone or as part of another group, refers to the group
"alkenyl-O--". For example, the term C.sub.2-10alkenyloxy means an
alkenyl group having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms and
at least one double bond bonded to the oxygen atom of the
alkenyloxy group. An exemplary alkoxy group is an allyloxy
group.
[0091] The term "alkynyloxy" as used herein, whether it is used
alone or as part of another group, refers to the group
"alkynyl-O--". For example, the term C.sub.2-10alkynyloxy means an
alkynyl group having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms and
at least one triple bond bonded to the oxygen atom of the
alkynyloxy group. An exemplary alkoxy group is a propargyloxy
group.
[0092] The term "aryloxy" as used herein, whether it is used alone
or as part of another group, refers to the group "aryl-O--". In an
embodiment of the present disclosure, the aryl group contains 6, 9,
10 or 14 atoms such as phenyl, naphthyl, indanyl or
anthracenyl.
[0093] The term "cycloalkyl," as used herein, whether it is used
alone or as part of another group, means saturated alkyl groups
having at least one cyclic ring. For example, the term
C.sub.3-10cycloalkyl means a cycloalkyl group having 3, 4, 5, 6, 7,
8, 9 or 10 carbon atoms.
[0094] The term "cycloalkylene" as used herein refers to a
cycloalkyl group that contains substituents on two of its ends.
[0095] The term "aryl" as used herein, whether it is used alone or
as part of another group, refers to cyclic groups that contain at
least one aromatic ring. In an embodiment of the application, the
aryl group contains from 6, 9, 10 or 14 atoms, such as phenyl,
naphthyl, indanyl or anthracenyl.
[0096] The term "arylene" as used herein refers to an aryl group
that contains substituents on two of its ends.
[0097] The term "heteroarylene" as used herein refers to a
heteroaryl group that contains substituents on two of its ends.
[0098] The term "heterocycloalkyl" as used herein, whether it is
used alone or as part of another group, refers to a non-aromatic,
ring-containing group having one or more multivalent heteroatoms,
independently selected from the group consisting of N, O and S, as
a part of the ring structure and including at least 3 and up to 20
atoms in the ring(s). Heterocycloalkyl groups are either saturated
or unsaturated (i.e. contain one or more double bonds) and may
contain more than one ring. When a heterocycloalkyl group contains
more than one ring, the rings may be fused, bridged, spiro
connected or linked by a single bond.
[0099] A first ring group being "fused" with a second ring group
means the first ring and the second ring share at least two
adjacent atoms there between.
[0100] A first ring group being "bridged" with a second ring group
means the first ring and the second ring share at least two
non-adjacent atoms there between.
[0101] A first ring group being "spiro connected" with a second
ring group means the first ring and the second ring share one atom
there between.
[0102] Heterocycloalkyl includes monocyclic heterocycloalkyls such
as but not limited to aziridinyl, oxiranyl, thiiranyl, azetidinyl,
oxetanyl, thietanyl, pyrrolidinyl, pyrrolinyl, imidazolidinyl,
pyrazolidinyl, pyrazolinyl, dioxolanyl, sulfolanyl,
2,3-dihydrofuranyl, 2,5-dihydrofuranyl, tetrahydrofuranyl,
thiophanyl, piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,
morpholinyl, thiomorpholinyl, pyranyl, thiopyranyl,
2,3-dihydropyranyl, tetrahydropyranyl, 1,4-dihydropyridinyl,
1,4-dioxanyl, 1,3-dioxanyl, dioxanyl, homopiperidinyl,
2,3,4,7-tetrahydro-1H-azepinyl, homopiperazinyl, 1,3-dioxepanyl,
4,7-dihydro-1,3-dioxepinyl, and hexamethylene oxidyl. Additionally,
heterocycloalkyl includes polycyclic heterocycloalkyls such as but
not limited to pyrolizidinyl, and quinolizidinyl. In addition to
the polycyclic heterocycloalkyls described above, heterocycloalkyl
includes polycyclic heterocycloalkyls wherein the ring fusion
between two or more rings includes more than one bond common to
both rings and more than two atoms common to both rings. Examples
of such bridged heterocycles include but are not limited to
quinuclidinyl, diazabicyclo[2.2.1]heptyl and
7-oxabicyclo[2.2.1]heptyl.
[0103] The term "heteroaryl" as used herein means a monocyclic ring
or a polycyclic ring system containing 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, 15, 16, 17, 18, 19 or 20 atoms, of which one or more, for
example 1 to 8, 1 to 6, 1 to 5, or 1 to 4, of the atoms are a
heteromoiety selected from O, S, NH and NC.sub.1-6alkyl, with the
remaining atoms being C, CH or CH.sub.2, said ring system
containing at least one aromatic ring.
[0104] Heteroaryl includes for example, pyridinyl, pyrazinyl,
pyrimidinyl, pyridazinyl, thienyl, furyl, furazanyl, pyrrolyl,
imidazolyl, thiazolyl, oxazolyl, pyrazolyl, isothiazolyl,
isoxazolyl, 1,2,3-triazolyl, tetrazolyl, 1,2,3-thiadiazolyl,
1,2,3-oxadiazolyl, 1,2,4-triazolyl, 1,2,4-thiadiazolyl,
1,2,4-oxadiazolyl, 1,3,4-triazolyl, 1,3,4-thiadiazolyl and 1,3,4
oxadiazolyl.
[0105] Heteroaryl also includes polycyclic heteroaryls such as but
not limited to indolyl, indolinyl, isoindolinyl, quinolinyl,
tetrahydroquinolinyl, isoquinolinyl, tetrahydroisoquinolinyl,
1,4-benzodioxanyl, coumarinyl, dihydrocoumarinyl, benzofuranyl,
2,3-dihydrobenzofuranyl, isobenzofuranyl, chromenyl, chromanyl,
isochromanyl, xanthenyl, phenoxathiinyl, thianthrenyl, indolizinyl,
isoindolyl, indazolyl, purinyl, phthalazinyl, naphthyridinyl,
quinoxalinyl, quinazolinyl, cinnolinyl, pteridinyl,
phenanthridinyl, perimidinyl, phenanthrolinyl, phenazinyl,
phenothiazinyl, phenoxazinyl, 1,2-benzisoxazolyl, benzothiophenyl,
benzoxazolyl, benzthiazolyl, benzimidazolyl, benztriazolyl,
thioxanthinyl, carbazolyl, carbolinyl and acridinyl.
[0106] A five-membered heteroaryl is a heteroaryl with a ring
having five ring atoms, where 1, 2 or 3 ring atoms are a
heteromoiety selected from O, S, NH and NC.sub.1-6alkyl, Exemplary
five-membered heteroaryls include but are not limited to thienyl,
furyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, pyrazolyl,
isothiazolyl, isoxazolyl, 1,2,3-triazolyl, tetrazolyl,
1,2,3-thiadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-triazolyl,
1,2,4-thiadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-triazolyl,
1,3,4-thiadiazolyl, and 1,3,4-oxadiazolyl.
[0107] A six-membered heteroaryl is a heteroaryl with a ring having
six ring atoms wherein 1, 2 or 3 ring atoms are a heteromoiety
selected from O, S, NH and NC.sub.1-6alkyl, Exemplary six-membered
heteroaryls include but are not limited to pyridyl, pyrazinyl,
pyrimidinyl, triazinyl and pyridazinyl.
[0108] The term "cyclic moiety" as used herein refers to any
cycloalkyl, aryl, heteroaryl or heterocycloalkyl group as defined
herein.
[0109] The term "heteromoiety" as used herein refers to a group of
atoms containing at least one heteratom.
[0110] As a prefix, the term "substituted" as used herein refers to
a structure, molecule or group in which one or more available
hydrogen atoms are replaced with one or more other chemical groups.
In an embodiment, the chemical group is a C.sub.1-4alkyl. In
another embodiment, the chemical group is a C.sub.1-12alkyl or a
chemical group that contains one or more heteroatoms selected from
N, O, S, F, Cl, Br, I, and P. Exemplary chemical groups containing
one or more heteroatoms include heterocycloalkyl, heteroaryl,
--NO.sub.2, --OR, --R'OR, --Cl, --Br, --I, --F, --CF.sub.3,
--C(O)R, --NR.sub.2, --SR, --SO.sub.2R, --S(O)R, --CN, --C(O)OR,
--C(O)NR.sub.2, --NRC(O)R, --NRC(O)OR, --R'NR.sub.2, oxo (O), imino
(.dbd.NR), thio (.dbd.S), and oximino (.dbd.N--OR), wherein each
"R" is hydrogen or a C.sub.1-12alkyl and "R'" is a
C.sub.1-12alkylene. For example, substituted phenyl may refer to
nitrophenyl, pyridylphenyl, methoxyphenyl, chlorophenyl,
aminophenyl, etc., wherein the nitro, pyridyl, methoxy, chloro, and
amino groups may replace any available hydrogen on the phenyl
ring.
[0111] As a suffix, the term "substituted" as used herein in
relation to a first structure, molecule or group, followed by one
or more variables or names of chemical groups, refers to a second
structure, molecule or group that results from replacing one or
more available hydrogen atoms of the first structure, molecule or
group with the one or more variables or named chemical groups. For
example, a "phenyl substituted by nitro" refers to nitrophenyl.
[0112] The term "available hydrogen atoms" as used herein refers to
hydrogen atoms on a molecule or group that can be replaced with
another group under conditions that will not degrade or decompose
the parent compound. Such conditions include the use of protecting
groups to protect sensitive functional groups in the molecule while
the hydrogen atom is being replaced.
[0113] The term "optionally substituted" refers to groups,
structures, or molecules that are either substituted or
unsubstituted.
[0114] The term "amine" or "amino," as used herein, whether it is
used alone or as part of another group, refers to radicals of the
general formula --NRR', wherein R and R' are each independently
selected from hydrogen or a alkyl group, for example
C.sub.1-6alkyl.
[0115] The term "halo" as used herein refers to a halogen atom and
includes fluoro, chloro, bromo and iodo.
[0116] The term "acac" as used herein refers to
acetylacetonate.
[0117] The term "atm" as used herein refers to atmosphere.
[0118] The term "aq." as used herein refers to aqueous.
[0119] The terms "Boc" and "t-Boc" as used herein refers to the
group tert-butoxycarbonyl.
[0120] DCM as used herein refers to dichloromethane.
[0121] DIPEA as used herein refers to N,N-diisopropyl
ethylamine
[0122] DMF as used herein refers to dimethylformamide.
[0123] DMSO as used herein refers to dimethylsulfoxide.
[0124] EDCl.HCl as used herein refers to
N-[3-(dimethylamino)propyl]-N'-ethylcarbodiimide hydrochloride.
[0125] EDC as used herein refers to
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide.
[0126] Et.sub.2O as used herein refers to diethylether.
[0127] EtOAc as used herein refers to ethyl acetate.
[0128] Et as used herein refers to the group ethyl.
[0129] Fmoc as used herein refers to the group
9-fluorenylmethyloxycarbonyl.
[0130] The term "hr(s)" as used herein refers to hour(s).
[0131] The term "min(s)" as used herein refers to minute(s).
[0132] HOBt as used herein refers to N-hydroxybenzotriazole.
[0133] HBTU as used herein refers to
O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate.
[0134] MeOH as used herein refers to methanol.
[0135] Me as used herein refers to the group methyl.
[0136] t-BuLi as used herein refers to tert-butyllithium.
[0137] ON as used herein refers to overnight.
[0138] RT as used herein refers to room temperature.
[0139] TEA as used herein refers to triethylamine.
[0140] TFA as used herein refers to trifluoroacetic acid.
[0141] THF as used herein refers to tetrahydrofuran.
[0142] t-Bu as used herein refers to the group tertiary butyl.
[0143] SPE as used herein refers to solid phase extraction, for
example using columns containing silica gel for
mini-chromatography.
[0144] The term "protecting group" or "PG" and the like as used
herein refers to a chemical moiety which protects or masks a
reactive portion of a molecule to prevent side reactions in those
reactive portions of the molecule, while manipulating or reacting a
different portion of the molecule. After the manipulation or
reaction is complete, the protecting group is removed under
conditions that do not degrade or decompose the remaining portions
of the molecule. The selection of a suitable protecting group can
be made by a person skilled in the art. Many conventional
protecting groups are known in the art, for example as described in
"Protective Groups in Organic Chemistry" McOmie, J. F. W. Ed.,
Plenum Press, 1973, in Greene, T. W. and Wuts, P. G. M.,
"Protective Groups in Organic Synthesis", John Wiley & Sons,
3.sup.rd Edition, 1999 and in Kocienski, P. Protecting Groups, 3rd
Edition, 2003, Georg Thieme Verlag (The Americas). Examples of
suitable protecting groups include, but are not limited to t-Boc,
cbz, Ac, Ts, Ms, silyl ethers such as TMSi, TBDMS, TBDPS, Tf, Ns,
Bn, Fmoc, benzoyl, dimethoxytrityl, methoxyethoxymethyl ether,
methoxymethyl ether, pivaloyl, p-methyoxybenzyl ether,
tetrahydropyranyl, trityl, ethoxyethyl ethers, carbobenzyloxy,
benzoyl and the like.
[0145] Cbz as used herein refers to the group carboxybenzyl.
[0146] Ac as used herein refers to the group acetyl.
[0147] Ts (tosyl) as used herein refers to the group
p-toluenesulfonyl.
[0148] Ms as used herein refers to the group methanesulfonyl.
[0149] TMS as used herein refers to tetramethylsilane.
[0150] TMSi as used herein refers to the group trimethylsilyl.
[0151] TBDMS as used herein refers to the group
t-butyldimethylsilyl.
[0152] TBDPS as used herein refers to the group
t-butyldiphenylsilyl.
[0153] Tf as used herein refers to the group
trifluoromethanesulfonyl.
[0154] Ns as used herein refers to the group naphthalene
sulphonyl.
[0155] Bn as used herein refers to the group benzyl.
[0156] The term "cell" as used herein refers to a single cell or a
plurality of cells and includes a cell either in a cell culture or
in a subject.
[0157] The term "subject" as used herein includes all members of
the animal kingdom including mammals, and suitably refers to
humans. Thus the methods and uses of the present application are
applicable to both human therapy and veterinary applications. In an
embodiment, the subject is a mammal. In another embodiment, the
subject is human.
[0158] The term "pharmaceutically acceptable" means compatible with
the treatment of subjects, for example humans.
[0159] The term "pharmaceutically acceptable carrier" means a
non-toxic solvent, dispersant, excipient, adjuvant or other
material which is mixed with the active ingredient in order to
permit the formation of a pharmaceutical composition, i.e., a
dosage form capable of administration to a subject. One
non-limiting example of such a carrier is a pharmaceutically
acceptable oil typically used for parenteral administration.
[0160] The term "pharmaceutically acceptable salt" means either an
acid addition salt or a base addition salt which is suitable for,
or compatible with the treatment of subjects.
[0161] An acid addition salt suitable for, or compatible with, the
treatment of subjects is any non-toxic organic or inorganic acid
addition salt of any basic compound. Basic compounds that form an
acid addition salt include, for example, compounds comprising an
amine group. Illustrative inorganic acids which form suitable salts
include hydrochloric, hydrobromic, sulfuric, nitric and phosphoric
acids, as well as acidic metal salts such as sodium monohydrogen
orthophosphate and potassium hydrogen sulfate. Illustrative organic
acids which form suitable salts include mono-, di- and
tricarboxylic acids. Illustrative of such organic acids are, for
example, acetic, trifluoroacetic, propionic, glycolic, lactic,
pyruvic, malonic, succinic, glutaric, fumaric, malic, tartaric,
citric, ascorbic, maleic, hydroxymaleic, benzoic, hydroxybenzoic,
phenylacetic, cinnamic, mandelic, salicylic, 2-phenoxybenzoic,
p-toluenesulfonic acid and other sulfonic acids such as
methanesulfonic acid, ethanesulfonic acid and
2-hydroxyethanesulfonic acid. Either the mono- or di-acid salts can
be formed, and such salts can exist in either a hydrated, solvated
or substantially anhydrous form. In general, acid addition salts
are more soluble in water and various hydrophilic organic solvents,
and generally demonstrate higher melting points in comparison to
their free base forms. The selection criteria for the appropriate
salt will be known to one skilled in the art. Other
non-pharmaceutically acceptable salts such as but not limited to
oxalates may be used, for example in the isolation of compounds of
the application for laboratory use, or for subsequent conversion to
a pharmaceutically acceptable acid addition salt. In another
embodiment of the present invention, the compound of Formula I is
converted to a pharmaceutically acceptable salt or solvate thereof,
in particular an acid addition salt such as a hydrochloride,
hydrobromide, phosphate, acetate, fumarate, maleate, tartrate,
citrate, methanesulphonate or p-toluenesulphonate
[0162] A base addition salt suitable for, or compatible with, the
treatment of subjects is any non-toxic organic or inorganic base
addition salt of any acidic compound. Acidic compounds that form a
basic addition salt include, for example, compounds comprising a
carboxylic acid group. Illustrative inorganic bases which form
suitable salts include lithium, sodium, potassium, calcium,
magnesium or barium hydroxide as well as ammonia. Illustrative
organic bases which form suitable salts include aliphatic,
alicyclic or aromatic organic amines such as isopropylamine,
methylamine, trimethylamine, picoline, diethylamine, triethylamine,
tripropylamine, ethanolamine, 2-dimethylaminoethanol,
2-diethylaminoethanol, dicyclohexylamine, lysine, arginine,
histidine, caffeine, procaine, hydrabamine, choline, betaine,
ethylenediamine, glucosamine, methylglucamine, theobromine,
purines, piperazine, piperidine, N-ethylpiperidine, polyamine
resins, and the like. Exemplary organic bases are isopropylamine,
diethylamine, ethanolamine, trimethylamine, dicyclohexylamine,
choline, and caffeine. [See, for example, S. M. Berge, et al.,
"Pharmaceutical Salts," J. Pharm. Sci. 1977, 66, 1-19]. The
selection of the appropriate salt may be useful so that an ester
functionality, if any, elsewhere in a compound is not hydrolyzed.
The selection criteria for the appropriate salt will be known to
one skilled in the art.
[0163] Prodrugs of the compounds of the present application may be,
for example, conventional esters formed with available hydroxy,
thiol, amino or carboxyl groups. For example, available hydroxy or
amino groups may be acylated using an activated acid in the
presence of a base, and optionally, in inert solvent (e.g. an acid
chloride in pyridine). Some common esters which have been utilized
as prodrugs are phenyl esters, aliphatic (C.sub.1-C.sub.24) esters,
acyloxymethyl esters, carbamates and amino acid esters.
[0164] The term "solvate" as used herein means a compound, or a
salt or prodrug of a compound, wherein molecules of a suitable
solvent are incorporated in the crystal lattice. A suitable solvent
is physiologically tolerable at the dosage administered. Examples
of suitable solvents are ethanol, water and the like. When water is
the solvent, the molecule is referred to as a "hydrate". The
formation of solvates of the compounds of the application will vary
depending on the compound and the solvate. In general, solvates are
formed by dissolving the compound in the appropriate solvent and
isolating the solvate by cooling or using an antisolvent. The
solvate is typically dried or azeotroped under ambient conditions.
The selection of suitable conditions to form a particular solvate
can be made by a person skilled in the art.
[0165] The term "treating" or "treatment" as used herein and as is
well understood in the art, means an approach for obtaining
beneficial or desired results, including clinical results.
Beneficial or desired clinical results can include, but are not
limited to alleviation or amelioration of one or more symptoms or
conditions, diminishment of extent of disease, stabilized (i.e. not
worsening) state of disease, preventing spread of disease, delay or
slowing of disease progression, amelioration or palliation of the
disease state, diminishment of the reoccurrence of disease, and
remission (whether partial or total), whether detectable or
undetectable. "Treating" and "treatment" can also mean prolonging
survival as compared to expected survival if not receiving
treatment. "Treating" and "treatment" as used herein also include
prophylactic treatment. For example, a subject with early cancer
can be treated to prevent progression, or alternatively a subject
in remission can be treated with a compound or composition
described herein to prevent recurrence. Treatment methods comprise
administering to a subject a therapeutically effective amount of
one or more of the compounds of the application and optionally
consist of a single administration, or alternatively comprise a
series of administrations. For example, the compounds of the
application may be administered at least once a week. However, in
another embodiment, the compounds may be administered to the
subject from about one time per three weeks, or about one time per
week to about once daily for a given treatment. In another
embodiment, the compounds are administered 2, 3, 4, 5 or 6 times
daily. The length of the treatment period depends on a variety of
factors, such as the severity of the disease, disorder or
condition, the age of the subject, the concentration and/or the
activity of the compounds of the application, and/or a combination
thereof. It will also be appreciated that the effective dosage of
the compound used for the treatment may increase or decrease over
the course of a particular treatment regime. Changes in dosage may
result and become apparent by standard diagnostic assays known in
the art. In some instances, chronic administration may be required.
For example, the compounds are administered to the subject in an
amount and for a duration sufficient to treat the patient.
[0166] "Palliating" a disease or disorder means that the extent
and/or undesirable clinical manifestations of a disorder or a
disease state are lessened and/or time course of the progression is
slowed or lengthened, as compared to not treating the disorder.
[0167] The term "prevention" or "prophylaxis", or synonym thereto,
as used herein refers to a reduction in the risk or probability of
a patient becoming afflicted with a disease, disorder or condition
mediated by proteasome inhibition or manifesting a symptom
associated with a disease, disorder or condition mediated by
proteasome inhibition.
[0168] As used herein, the term "effective amount" or
"therapeutically effective amount" means an amount effective, at
dosages and for periods of time necessary to achieve the desired
result. For example in the context of treating a disease, disorder
or condition mediated by proteasome inhibition, an effective amount
is an amount that, for example, increases proteasome inhibition
compared to the proteasome inhibition without administration of the
compound. Effective amounts may vary according to factors such as
the disease state, age, sex and/or weight of the subject. The
amount of a given compound that will correspond to such an amount
will vary depending upon various factors, such as the given drug or
compound, the pharmaceutical formulation, the route of
administration, the type of condition, disease or disorder, the
identity of the subject being treated, and the like, but can
nevertheless be routinely determined by one skilled in the art.
[0169] The term "mediated by" or "associated with" as used herein
refers to a disease, disorder or condition in a subject wherein at
least one of the causes is the specified physiological abnormality,
for example an enhanced level of proteasome activity, in particular
compared to subjects that do not have the disease, disorder or
condition.
[0170] The term "administered" as used herein means administration
of a therapeutically effective amount of a compound or composition
of the application to a cell either in cell culture or in a
subject.
[0171] The term "neoplastic disorder" as used herein refers to a
disease, disorder or condition characterized by cells that have the
capacity for autonomous growth or replication, e.g., an abnormal
state or condition characterized by proliferative cell growth. The
term "neoplasm" as used herein refers to a mass of tissue resulting
from the abnormal growth and/or division of cells in a subject
having a neoplastic disorder. Neoplasms can be benign (such as
uterine fibroids and melanocytic nevi), potentially malignant (such
as carcinoma in situ) or malignant (i.e. cancer). Exemplary
neoplastic disorders include but are not limited to carcinoma,
sarcoma, metastatic disorders (e.g., tumors arising from the
prostate), hematopoietic neoplastic disorders, (e.g., leukemias,
lymphomas, myeloma and other malignant plasma cell disorders),
metastatic tumors and other cancers. Prevalent cancers include
breast, prostate, colon, lung, liver, brain, ovarian and pancreatic
cancers.
[0172] The term "cancer" as used herein refers to
cellular-proliferative disease states, including but not limited
to: Acute Lymphoblastic Leukemia, Adult; Acute Lymphoblastic
Leukemia, Childhood; Acute Myeloid Leukemia, Adult; Adrenocortical
Carcinoma; Adrenocortical Carcinoma, Childhood; AIDS-Related
Lymphoma; AIDS-Related Malignancies; Anal Cancer; Astrocytoma,
Childhood Cerebellar; Astrocytoma, Childhood Cerebral; Bile Duct
Cancer, Extrahepatic; Bladder Cancer; Bladder Cancer, Childhood;
Bone Cancer, Osteosarcoma/Malignant Fibrous Histiocytoma; Brain
Stem Glioma, Childhood; Brain Tumor, Adult; Brain Tumor, Brain Stem
Glioma, Childhood; Brain Tumor, Cerebellar Astrocytoma, Childhood;
Brain Tumor, Cerebral Astrocytoma/Malignant Glioma, Childhood;
Brain Tumor, Ependymoma, Childhood; Brain Tumor, Medulloblastoma,
Childhood; Brain Tumor, Supratentorial Primitive Neuroectodermal
Tumors, Childhood; Brain Tumor, Visual Pathway and Hypothalamic
Glioma, Childhood; Brain Tumor, Childhood (Other); Breast Cancer;
Breast Cancer and Pregnancy; Breast Cancer, Childhood; Breast
Cancer, Male; Bronchial Adenomas/Carcinoids, Childhood; Carcinoid
Tumor, Childhood; Carcinoid Tumor, Gastrointestinal; Carcinoma,
Adrenocortical; Carcinoma, Islet Cell; Carcinoma of Unknown
Primary; Central Nervous System Lymphoma, Primary; Cerebellar
Astrocytoma, Childhood; Cerebral Astrocytoma/Malignant Glioma,
Childhood; Cervical Cancer; Childhood Cancers; Chronic Lymphocytic
Leukemia; Chronic Myelogenous Leukemia; Chronic Myeloproliferative
Disorders; Clear Cell Sarcoma of Tendon Sheaths; Colon Cancer;
Colorectal Cancer, Childhood; Cutaneous T-Cell Lymphoma;
Endometrial Cancer; Ependymoma, Childhood; Epithelial Cancer,
Ovarian; Esophageal Cancer; Esophageal Cancer, Childhood; Ewing's
Family of Tumors; Extracranial Germ Cell Tumor, Childhood;
Extragonadal Germ Cell Tumor; Extrahepatic Bile Duct Cancer; Eye
Cancer, Intraocular Melanoma; Eye Cancer, Retinoblastoma;
Gallbladder Cancer; Gastric (Stomach) Cancer; Gastric (Stomach)
Cancer, Childhood; Gastrointestinal Carcinoid Tumor; Germ Cell
Tumor, Extracranial, Childhood; Germ Cell Tumor, Extragonadal; Germ
Cell Tumor, Ovarian; Gestational Trophoblastic Tumor; Glioma,
Childhood Brain Stem; Glioma, Childhood Visual Pathway and
Hypothalamic; Hairy Cell Leukemia; Head and Neck Cancer;
Hepatocellular (Liver) Cancer, Adult (Primary); Hepatocellular
(Liver) Cancer, Childhood (Primary); Hodgkin's Lymphoma, Adult;
Hodgkin's Lymphoma, Childhood; Hodgkin's Lymphoma During Pregnancy;
Hypopharyngeal Cancer; Hypothalamic and Visual Pathway Glioma,
Childhood; Intraocular Melanoma; Islet Cell Carcinoma (Endocrine
Pancreas); Kaposi's Sarcoma; Kidney Cancer; Laryngeal Cancer;
Laryngeal Cancer, Childhood; Leukemia, Acute Lymphoblastic, Adult;
Leukemia, Acute Lymphoblastic, Childhood; Leukemia, Acute Myeloid,
Adult; Leukemia, Acute Myeloid, Childhood; Leukemia, Chronic
Lymphocytic; Leukemia, Chronic Myelogenous; Leukemia, Hairy Cell;
Lip and Oral Cavity Cancer; Liver Cancer, Adult (Primary); Liver
Cancer, Childhood (Primary); Lung Cancer, Non-Small Cell; Lung
Cancer, Small Cell; Lymphoblastic Leukemia, Adult Acute;
Lymphoblastic Leukemia, Childhood Acute; Lymphocytic Leukemia,
Chronic; Lymphoma, AIDS-Related; Lymphoma, Central Nervous System
(Primary); Lymphoma, Cutaneous T-Cell; Lymphoma, Hodgkin's, Adult;
Lymphoma, Hodgkin's, Childhood; Lymphoma, Hodgkin's During
Pregnancy; Lymphoma, Non-Hodgkin's, Adult; Lymphoma, Non-Hodgkin's,
Childhood; Lymphoma, Non-Hodgkin's During Pregnancy; Lymphoma,
Primary Central Nervous System; Macroglobulinemia, Waldenstrom's;
Male Breast Cancer; Malignant Mesothelioma, Adult; Malignant
Mesothelioma, Childhood; Malignant Thymoma; Medulloblastoma,
Childhood; Melanoma; Melanoma, Intraocular; Merkel Cell Carcinoma;
Mesothelioma, Malignant; Metastatic Squamous Neck Cancer with
Occult Primary; Multiple Endocrine Neoplasia Syndrome, Childhood;
Multiple Myeloma/Plasma Cell Neoplasm; Mycosis Fungoides;
Myelodysplastic Syndromes; Myelogenous Leukemia, Chronic; Myeloid
Leukemia, Childhood Acute; Myeloma, Multiple; Myeloproliferative
Disorders, Chronic; Nasal Cavity and Paranasal Sinus Cancer;
Nasopharyngeal Cancer; Nasopharyngeal Cancer, Childhood;
Neuroblastoma; Non-Hodgkin's Lymphoma, Adult; Non-Hodgkin's
Lymphoma, Childhood; Non-Hodgkin's Lymphoma During Pregnancy;
Non-Small Cell Lung Cancer; Oral Cancer, Childhood; Oral Cavity and
Lip Cancer; Oropharyngeal Cancer; Osteosarcoma/Malignant Fibrous
Histiocytoma of Bone; Ovarian Cancer, Childhood; Ovarian Epithelial
Cancer; Ovarian Germ Cell Tumor; Ovarian Low Malignant Potential
Tumor; Pancreatic Cancer; Pancreatic Cancer, Childhood; Pancreatic
Cancer, Islet Cell; Paranasal Sinus and Nasal Cavity Cancer;
Parathyroid Cancer; Penile Cancer; Pheochromocytoma; Pineal and
Supratentorial Primitive Neuroectodermal Tumors, Childhood;
Pituitary Tumor; Plasma Cell Neoplasm/Multiple Myeloma;
Pleuropulmonary Blastoma; Pregnancy and Breast Cancer; Pregnancy
and Hodgkin's Lymphoma; Pregnancy and Non-Hodgkin's Lymphoma;
Primary Central Nervous System Lymphoma; Primary Liver Cancer,
Adult; Primary Liver Cancer, Childhood; Prostate Cancer; Rectal
Cancer; Renal Cell (Kidney) Cancer; Renal Cell Cancer, Childhood;
Renal Pelvis and Ureter, Transitional Cell Cancer; Retinoblastoma;
Rhabdomyosarcoma, Childhood; Salivary Gland Cancer; Salivary Gland
Cancer, Childhood; Sarcoma, Ewing's Family of Tumors; Sarcoma,
Kaposi's; Sarcoma (Osteosarcoma)/Malignant Fibrous Histiocytoma of
Bone; Sarcoma, Rhabdomyosarcoma, Childhood; Sarcoma, Soft Tissue,
Adult; Sarcoma, Soft Tissue, Childhood; Sezary Syndrome; Skin
Cancer; Skin Cancer, Childhood; Skin Cancer (Melanoma); Skin
Carcinoma, Merkel Cell; Small Cell Lung Cancer; Small Intestine
Cancer; Soft Tissue Sarcoma, Adult; Soft Tissue Sarcoma, Childhood;
Squamous Neck Cancer with Occult Primary, Metastatic; Stomach
(Gastric) Cancer; Stomach (Gastric) Cancer, Childhood;
Supratentorial Primitive Neuroectodermal Tumors, Childhood; T-Cell
Lymphoma, Cutaneous; Testicular Cancer; Thymoma, Childhood;
Thymoma, Malignant; Thyroid Cancer; Thyroid Cancer, Childhood;
Transitional Cell Cancer of the Renal Pelvis and Ureter;
Trophoblastic Tumor, Gestational; Unknown Primary Site, Cancer of,
Childhood; Unusual Cancers of Childhood; Ureter and Renal Pelvis,
Transitional Cell Cancer; Urethral Cancer; Uterine Sarcoma; Vaginal
Cancer; Visual Pathway and Hypothalamic Glioma, Childhood; Vulvar
Cancer; Waldenstrom's Macro globulinemia; and Wilms' Tumor.
Metastases of the aforementioned cancers can also be treated in
accordance with the methods described herein.
II. Compounds of the Application
[0173] Compounds of the present application were prepared and found
to inhibit uncontrolled and/or abnormal cellular activities
affected directly or indirectly by the proteasome. In particular,
compounds of the present application exhibited activity as
proteasome inhibitors, and are therefore useful in therapy, for
example for the treatment of neoplastic disorders such as cancer
and neurodegenerative disorders associated directly or indirectly
with proteasome inhibition.
[0174] Accordingly, the present application includes a compound of
Formula I or a pharmaceutically acceptable salt, solvate or prodrug
thereof:
##STR00003##
wherein:
[0175] R.sup.1 is selected from the group consisting of
C.sub.1-10alkyl, C.sub.2-10alkenyl, C.sub.2-10alkynyl,
C.sub.1-10haloalkyl, C.sub.1-10cyanoalkyl, C.sub.1-10alkoxy,
C.sub.2-10alkenyloxy, C.sub.2-10alkynyloxy, C.sub.3-10cycloalkyl,
heterocycloalkyl, aryl, heteroaryl,
C.sub.1-6alkylene-O--C.sub.1-6alkyl,
C.sub.1-6alkylene-O--C.sub.1-6haloalkyl,
C.sub.2-6alkenylene-O--C.sub.1-6haloalkyl,
C.sub.2-6alkynylene-O--C.sub.1-6haloalkyl,
C.sub.1-6alkylene-C.sub.3-8cycloalkyl,
C.sub.1-6alkylene-heterocycloalkyl, C.sub.1-6alkylene-aryl,
C.sub.1-6alkylene-heteroaryl, C(O)R.sup.7, OC(O)R.sup.7,
C(O)OR.sup.7, C.sub.1-6alkylene-O--R.sup.7,
C.sub.1-6alkylene-C(O)R.sup.7, C.sub.1-6alkylene-O--C(O)R.sup.7,
C.sub.1-6alkylene-C(O)OR.sup.7, C.sub.1-6alkylene-O--C(O)OR.sup.7,
C.sub.1-6alkylene-NR.sup.7R.sup.8,
C.sub.1-6alkylene-C(O)NR.sup.7R.sup.8,
C.sub.1-6alkylene-NR.sup.7C(O)R.sup.8,
C.sub.1-6alkylene-NR.sup.7C(O)NR.sup.7R.sup.8,
C.sub.1-6alkylene-S--R.sup.7, C.sub.1-6alkylene-S(O)R.sup.7,
C.sub.1-6alkylene-SO.sub.2R.sup.7,
C.sub.1-6alkylene-SO.sub.2NR.sup.7R.sup.8,
C.sub.1-6alkylene-NR.sup.7SO.sub.2R.sup.8,
C.sub.1-6alkylene-NR.sup.7SO.sub.2NR.sup.7R.sup.8,
C(O)NR.sup.7R.sup.8 and C.sub.1-6alkylene-NR.sup.7C(O)OR.sup.8,
wherein any cyclic moiety is optionally substituted with
C.sub.1-4alkyl and/or is optionally fused to a further cyclic
moiety;
[0176] X is absent or is selected from the group consisting of O,
NH, NC.sub.1-6alkyl, S, S(O), SO.sub.2, C(O), C.sub.1-6alkylene,
C.sub.2-6alkenylene, C.sub.2-6alkynylene, C.sub.1-6haloalkylene,
C.sub.3-8cycloalkylene, heterocycloalkylene, arylene and
heteroarylene, or X is a combination of two or three of O, NH,
NC.sub.1-6alkyl, S, S(O), SO.sub.2, C.sub.1-6alkylene,
C.sub.2-6alkenylene, C.sub.2-6alkynylene, C.sub.1-6haloalkylene,
C.sub.3-8cycloalkylene, heterocycloalkylene, arylene or
heteroarylene, bonded together in a linear fashion, provided that
two or three of O, NH, NC.sub.1-6alkyl, S, S(O) and SO.sub.2 and
not bonded directly to each other;
[0177] R.sup.2, R.sup.3 and R.sup.4 are each independently selected
from the group consisting of C.sub.1-10alkyl, C.sub.2-10alkenyl,
C.sub.2-10alkynyl, C.sub.1-10haloalkyl, C.sub.1-10cyanoalkyl,
C.sub.1-10alkoxy, C.sub.2-10alkenyloxy, C.sub.2-10alkynyloxy,
C.sub.3-10cycloalkyl, heterocycloalkyl, aryl, heteroaryl,
C.sub.1-6alkylene-O--C.sub.1-6alkyl,
C.sub.1-6alkylene-O--C.sub.1-6haloalkyl,
C.sub.2-6alkenylene-O--C.sub.1-6haloalkyl,
C.sub.2-6alkynylene-O--C.sub.1-6haloalkyl,
C.sub.1-6alkylene-C.sub.3-8cycloalkyl,
C.sub.1-6alkylene-heterocycloalkyl, C.sub.1-6alkylene-aryl,
C.sub.1-6alkylene-heteroaryl, C(O)R.sup.7, OC(O)R.sup.7,
C(O)OR.sup.7, C.sub.1-6alkylene-O--R.sup.7,
C.sub.1-6alkylene-C(O)R.sup.7, C.sub.1-6alkylene-O--C(O)R.sup.7,
C.sub.1-6alkylene-C(O)OR.sup.7, C.sub.1-6alkylene-O--C(O)OR.sup.7,
C.sub.1-6alkylene-NR.sup.7R.sup.8,
C.sub.1-6alkylene-C(O)NR.sup.7R.sup.8,
C.sub.1-6alkylene-NR.sup.7C(O)R.sup.8,
C.sub.1-6alkylene-NR.sup.7C(O)NR.sup.7R.sup.8,
C.sub.1-6alkylene-S--R.sup.7, C.sub.1-6alkylene-S(O)R.sup.7,
C.sub.1-6alkylene-SO.sub.2R.sup.7,
C.sub.1-6alkylene-SO.sub.2NR.sup.7R.sup.8,
C.sub.1-6alkylene-NR.sup.7SO.sub.2R.sup.8,
C.sub.1-6alkylene-NR.sup.7SO.sub.2NR.sup.7R.sup.8,
C(O)NR.sup.7R.sup.8 and C.sub.1-6alkylene-NR.sup.7C(O)OR.sup.8,
wherein any cyclic moiety is optionally fused to a further 5- to
7-membered cyclic moiety, wherein at least one of R.sup.2, R.sup.3
and R.sup.4 is C.sub.1-6-alkylene-O--C.sub.1-6haloalkyl, and
wherein R.sup.2, R.sup.3 and R.sup.4 are optionally substituted
with one or more independently-selected R.sup.6 groups;
[0178] R.sup.5 is selected from the group consisting of H,
C.sub.1-6alkyl, C.sub.1-6haloalkyl, C.sub.2-6alkenyl,
C.sub.2-6alkynyl, C.sub.3-8cycloalkyl,
C.sub.1-6alkylene-C.sub.3-8cycloalkyl, aryl, heteroaryl, and
heterocycloalkyl;
[0179] R.sup.6 is selected from the group consisting of
C.sub.1-6alkyl, OH, halo, O--(C.sub.2-3alkylene)-O,
C.sub.1-6alkoxy, aryloxy, --NH--C.sub.1-6alkyl,
--N(C.sub.1-6alkyl).sub.2,
C.sub.1-6alkylene-N(C.sub.1-6alkyl).sub.2,
C.sub.1-6alkylene-NH--C.sub.1-6alkyl, cycloalkyl, heterocycloalkyl,
aryl and heteroaryl; and
[0180] R.sup.7 and R.sup.8 are each independently selected from the
group consisting of H, C.sub.1-6alkyl, C.sub.1-6haloalkyl,
C.sub.2-6alkenyl, C.sub.2-6alkynyl, C.sub.3-10cycloalkyl,
C.sub.1-6alkylene-C.sub.3-10cycloalkyl, heterocycloalkyl, aryl,
C.sub.1-6alkylene-aryl, C.sub.1-6alkylene-heterocycloalkyl,
heteroaryl, and C.sub.1-6alkylene-heteroaryl, wherein any cyclic
moiety is optionally fused to a further cyclic moiety.
[0181] In an embodiment, R.sup.1 is selected from: [0182] (i)
C.sub.1-6alkyl; [0183] (ii) C.sub.2-10alkenyl; [0184] (iii)
C.sub.2-10alkynyl; [0185] (iv) substituted or unsubstituted
C.sub.6-14aryl; [0186] (v) substituted or unsubstituted heteroaryl;
[0187] (vi) substituted or unsubstituted C.sub.3-10cycloalkyl; and
[0188] (vii) substituted or unsubstituted
C.sub.3-10heterocycloalkyl, wherein the substituents for
C.sub.6-14aryl, heteroaryl, C.sub.3-10cycloalkyl and
C.sub.3-10heterocycloalkyl are independently selected from
C.sub.1-4alkyl.
[0189] In another embodiment, R.sup.1 is selected from: [0190] (i)
C.sub.1-6alkyl; [0191] (ii) C.sub.2-6alkenyl; [0192] (iii)
C.sub.2-6alkynyl; [0193] (iv) substituted or unsubstituted
C.sub.6-10aryl; [0194] (v) substituted or unsubstituted 5- or
6-membered heteroaryl; [0195] (vi) substituted or unsubstituted
C.sub.3-8cycloalkyl; and [0196] (vii) substituted or unsubstituted
C.sub.3-8heterocycloalkyl, wherein the substituents for
C.sub.6-10aryl, 5- or 6-membered heteroaryl, C.sub.3-8cycloalkyl
and C.sub.3-8heterocycloalkyl are independently selected from
C.sub.1-4alkyl.
[0197] In a further embodiment, R.sup.1 is selected from: [0198]
(i) C.sub.1-6alkyl; [0199] (ii) a substituted or unsubstituted 5-
or 6-membered heteroaryl; and [0200] (iii)
C.sub.3-8heterocycloalkyl, wherein the substituents for the 5- or
6-membered heteroaryl are independently selected from
C.sub.1-4alkyl.
[0201] It is an embodiment that R.sup.1 is selected from the group
consisting of an unsubstituted 5- or 6-membered heteroaryl, a 5- or
6-membered heteroaryl substituted with a C.sub.1-4alkyl and
C.sub.3-8heterocycloalkyl.
[0202] In an embodiment, R.sup.1 is C.sub.1-10alkyl. In another
embodiment, R.sup.1 is C.sub.1-6alkyl. In a further embodiment,
R.sup.1 is t-butyl.
[0203] It is an embodiment that R.sup.1 is an unsubstituted 5- or
6-membered heteroaryl, a 5-membered heteroaryl substituted with a
C.sub.1-4alkyl or a 6-membered heterocycloalkyl.
[0204] In an embodiment, R.sup.1 is heterocycloalkyl. In another
embodiment, R.sup.1 is selected from morpholinyl, 1,4-oxazepanyl,
thiomorpholinyl, 1,4-thiazepanyl, 1,4-thiazepanyl-1-oxide,
1,4-thiazepanyl-1,1-dioxide, 1,4-thiazinanyl-1-oxide,
1,4-thiazinanyl-1,1-dioxide, aziridinyl, azetidinyl, pyrrolidinyl,
piperazinyl and 1,4-diazepanyl. In another embodiment, R.sup.1 is a
6-membered heterocycloalkyl having one O atom and one N atom as a
part of the ring structure. It is an embodiment that R.sup.1 is
morpholinyl. In another embodiment of the present application,
R.sup.1 is
##STR00004##
[0205] In an embodiment, R.sup.1 is an unsubstituted 5- or
6-membered heteroaryl or a 5- or 6-membered heteroaryl substituted
with a C.sub.1-4alkyl. In another embodiment, R.sup.1 is an
unsubstituted 5-membered heteroaryl. In a further embodiment,
R.sup.1 is an unsubstituted 6-membered heteroaryl. It is an
embodiment that R.sup.1 is a 5-membered heteroaryl, substituted
with a C.sub.1-4alkyl. In another embodiment, R.sup.1 is a
5-membered heteroaryl, substituted with a methyl.
[0206] In an embodiment, R.sup.1 is selected from: [0207] (i)
substituted or unsubstituted thiazolyl; [0208] (ii) substituted or
unsubstituted isothiazolyl; [0209] (iii) substituted or
unsubstituted oxazolyl; [0210] (iv) substituted or unsubstituted
isooxazolyl; [0211] (v) substituted or unsubstituted thiophenyl;
[0212] (vi) substituted or unsubstituted furanyl; [0213] (vii)
substituted or unsubstituted 1,2,4-triazolyl; [0214] (viii)
substituted or unsubstituted pyridyl; [0215] (ix) substituted or
unsubstituted pyrazinyl; [0216] (x) substituted or unsubstituted
pyrimidinyl; and [0217] (xi) substituted or unsubstituted
1,2,4-triazinyl, wherein the substituents for thiazolyl,
isothiazolyl, oxazolyl, isooxazolyl, thiophenyl, furanyl,
1,2,4-triazolyl, pyridyl, pyrazinyl, pyrimidinyl and
1,2,4-triazinyl are independently selected from a C.sub.1-4alkyl
such as a methyl.
[0218] In another embodiment R.sup.1 is selected from: [0219] (i)
substituted or unsubstituted isoxazolyl; [0220] (ii) substituted or
unsubstituted isothiazolyl; [0221] (iii) substituted or
unsubstituted furanyl; [0222] (iv) substituted or unsubstituted
thiophenyl; [0223] (v) substituted or unsubstituted oxazolyl;
[0224] (vi) substituted or unsubstituted thiazolyl; [0225] (vii)
substituted or unsubstituted pyrazolyl; and [0226] (viii)
substituted or unsubstituted imidazolyl, wherein the substituents
for isoxazolyl, isothiazolyl, furanyl, thiophenyl, oxazolyl,
thiazolyl, pyrazolyl and imidazolyl are independently selected from
a C.sub.1-4alkyl such as a methyl.
[0227] In another embodiment, R.sup.1 is selected from: [0228] (i)
substituted or unsubstituted isoxazolyl; [0229] (ii) substituted or
unsubstituted furanyl; and [0230] (iii) substituted or
unsubstituted thiazolyl, wherein the substituents for isooxazolyl,
furanyl and thiazolyl are independently selected from a
C.sub.1-4alkyl such as a methyl.
[0231] In another embodiment, R.sup.1 is selected from:
##STR00005##
[0232] In an embodiment, X is absent or is selected from O, NH,
NC.sub.1-6alkyl, S, C.sub.1-6alkylene, C.sub.2-6alkenylene and
C.sub.2-6alkynylene. In another embodiment, X is absent. In a
further embodiment, X is O or C.sub.1-6alkylene. It is an
embodiment that X is O. In another embodiment, X is
C.sub.1-4alkylene. In a further embodiment X is --CH.sub.2--.
[0233] In an embodiment, X is O and R.sup.1 is C.sub.1-6alkyl. In
another embodiment, X is O and R.sup.1 is t-butyl.
[0234] In another embodiment, X is C.sub.1-6alkylene and R.sup.1 is
C.sub.3-8heterocycloalkyl. In a further embodiment, X is
C.sub.1-4alkylene and R.sup.1 is a 6-membered heterocycloalkyl. It
is an embodiment of the present application that X and R.sup.1
together form the structure:
##STR00006##
[0235] In other embodiments, X is absent and R.sup.1 is substituted
or unsubstituted heteroaryl, wherein the substituents for
heteroaryl are independently selected from a C.sub.1-4alkyl such as
a methyl. It will be appreciated that in such embodiments of the
present application, the heteroaryl embodiments can be as discussed
above in respect of R.sup.1.
[0236] In an embodiment, R.sup.2, R.sup.3 and R.sup.4 are each
independently selected from the group consisting of
C.sub.1-10alkyl, C.sub.2-10alkenyl, C.sub.2-10alkynyl,
C.sub.1-6alkyleneC.sub.6-14aryl, C.sub.1-6alkylene-heteroaryl,
C.sub.1-6alkyleneC.sub.3-8cycloalkyl,
C.sub.1-6alkylene-O--C.sub.1-6alkyl,
C.sub.1-6alkylene-O--C.sub.1-6haloalkyl,
C.sub.2-6alkenylene-O--C.sub.1-6haloalkyl and
C.sub.2-6alkynylene-O--C.sub.1-6haloalkyl, wherein at least one of
R.sup.2, R.sup.3 and R.sup.4 is
C.sub.1-6-alkylene-O--C.sub.1-6haloalkyl. In another embodiment,
R.sup.2, R.sup.3 and R.sup.4 are each independently selected from
the group consisting of C.sub.1-6alkyl,
C.sub.1-6alkyleneC.sub.6-10aryl,
C.sub.1-6alkylene-O--C.sub.1-4alkyl and
C.sub.1-6alkylene-O--C.sub.1-4fluoroalkyl, wherein at least one of
R.sup.2, R.sup.3 and R.sup.4 is
C.sub.1-6alkylene-O--C.sub.1-4fluoroalkyl. In a further embodiment
R.sup.2, R.sup.3 and R.sup.4 are each independently selected from
the group consisting of C.sub.1-6alkyl, C.sub.1-4alkylene-phenyl,
C.sub.1-4alkylene-O--C.sub.1-4alkyl,
C.sub.1-4alkylene-O--CH.sub.2F, C.sub.1-4alkylene-O--CHF.sub.2 and
C.sub.1-4alkylene-O--CF.sub.3, wherein at least one of R.sup.2,
R.sup.3 and R.sup.4 is C.sub.1-4alkylene-O--CH.sub.2F,
C.sub.1-4alkylene-O--CHF.sub.2 or C.sub.1-4alkylene-O--CF.sub.3. It
is an embodiment that R.sup.2, R.sup.3 and R.sup.4 are each
independently selected from the group consisting of isobutyl,
--CH.sub.2-Ph, --CH.sub.2--O--CH.sub.3, --CH.sub.2--O--CH.sub.2F,
--CH.sub.2--O--CHF.sub.2 and --CH.sub.2--O--CF.sub.3, wherein at
least one of R.sup.2, R.sup.3 and R.sup.4 is
--CH.sub.2--O--CH.sub.2F, --CH.sub.2--O--CHF.sub.2 or
--CH.sub.2--O--CF.sub.3 In another embodiment, R.sup.2, R.sup.3 and
R.sup.4 are each independently selected from the group consisting
of isobutyl, --CH.sub.2-Ph, --CH.sub.2--O--CH.sub.3 and
--CH.sub.2--O--CHF.sub.2, wherein at least one of R.sup.2, R.sup.3
and R.sup.4 is --CH.sub.2--O--CHF.sub.2.
[0237] In an embodiment, R.sup.2 and R.sup.3 are each independently
selected from the group consisting of C.sub.1-10alkyl,
C.sub.2-10alkenyl, C.sub.2-10alkynyl,
C.sub.1-6alkylene-O--C.sub.1-6alkyl,
C.sub.1-6alkylene-O--C.sub.1-6haloalkyl,
C.sub.2-6alkenylene-O--C.sub.1-6haloalkyl and
C.sub.2-6alkynylene-O--C.sub.1-6haloalkyl, wherein at least one of
R.sup.2 and R.sup.3 is C.sub.1-6-alkylene-O--C.sub.1-6haloalkyl. In
another embodiment, R.sup.2 and R.sup.3 are each independently
selected from the group consisting of C.sub.1-6alkyl,
C.sub.1-6alkylene-O--C.sub.1-4alkyl and
C.sub.1-6alkylene-O--C.sub.1-4fluoroalkyl, wherein at least one of
R.sup.2 and R.sup.3 is C.sub.1-6alkylene-O--C.sub.1-4fluoroalkyl.
In a further embodiment R.sup.2 and R.sup.3 are each independently
selected from the group consisting of C.sub.1-6alkyl,
C.sub.1-4alkylene-O--C.sub.1-4alkyl,
C.sub.1-4alkylene-O--CH.sub.2F, C.sub.1-4alkylene-O--CHF.sub.2 and
C.sub.1-4alkylene-O--CF.sub.3 wherein at least one of R.sup.2 and
R.sup.3 is C.sub.1-4alkylene-O--CH.sub.2F,
C.sub.1-4alkylene-O--CHF.sub.2 or C.sub.1-4alkylene-O--CF.sub.3. It
is an embodiment that R.sup.2 and R.sup.3 are each independently
selected from the group consisting of isobutyl,
--CH.sub.2--O--CH.sub.3, --CH.sub.2--O--CH.sub.2F,
--CH.sub.2--O--CHF.sub.2 and --CH.sub.2--O--CF.sub.3, wherein at
least one of R.sup.2 and R.sup.3 is --CH.sub.2--O--CH.sub.2F,
--CH.sub.2--O--CHF.sub.2 or --CH.sub.2--O--CF.sub.3. In another
embodiment, R.sup.2 and R.sup.3 are each independently selected
from the group consisting of isobutyl, --CH.sub.2--O--CH.sub.3 and
--CH.sub.2--O--CHF.sub.2, wherein at least one of R.sup.2 and
R.sup.3 is --CH.sub.2--O--CHF.sub.2.
[0238] In an embodiment, R.sup.2 and R.sup.3 are each
C.sub.1-6alkylene-O--C.sub.1-4fluoroalkyl. In another embodiment,
R.sup.2 and R.sup.3 are each C.sub.1-4alkylene-O--CHF.sub.2. In a
further embodiment, R.sup.2 and R.sup.3 are each
--CH.sub.2--O--CHF.sub.2.
[0239] In an embodiment, R.sup.4 is selected from the group
consisting of C.sub.1-10alkyl, C.sub.2-10alkenyl,
C.sub.2-10alkynyl, C.sub.1-6alkyleneC.sub.3-8cycloalkyl and
C.sub.1-6alkyleneC.sub.6-14aryl. In another embodiment, R.sup.4 is
selected from the group consisting of C.sub.1-6alkyl,
C.sub.1-6alkyleneC.sub.3-8cycloalkyl, and
C.sub.1-6alkyleneC.sub.6-10aryl. In a further embodiment, R.sup.4
is selected from the group consisting of C.sub.1-6alkyl and
C.sub.1-4alkylene-phenyl. It is an embodiment that R.sup.4 is
isobutyl or --CH.sub.2-Ph. In an embodiment, R.sup.4 is isobutyl.
In another embodiment, R.sup.4 is --CH.sub.2-Ph.
[0240] In another embodiment, at least one of R.sup.2 and R.sup.3
or both is independently selected from the group consisting of
C.sub.1-6alkylene-O--CH.sub.2F, C.sub.1-6alkylene-O--CHF.sub.2 and
C.sub.1-6alkylene-O--CF.sub.3. In a further embodiment, at least
one of R.sup.2 and R.sup.3 or both is --CH.sub.2--O--CHF.sub.2.
[0241] In another embodiment, R.sup.4 is selected from
C.sub.1-6alkyl, C.sub.1-6alkyleneC.sub.3-8cycloalkyl and
C.sub.1-6alkyleneC.sub.6-14aryl, optionally substituted with one or
more independently-selected groups R.sup.6. In a further
embodiment, R.sup.4 is selected from --CH.sub.2-phenyl
--(CH.sub.2).sub.2-phenyl, isobutyl, and tert-butyl. It is an
embodiment that R.sup.4 is --CH.sub.2-phenyl or isobutyl.
[0242] In an embodiment, R.sup.5 is selected from the group
consisting of H, C.sub.1-6alkyl, C.sub.2-6alkenyl and
C.sub.2-6alkynyl. In another embodiment, R.sup.5 is C.sub.1-6alkyl.
In a further embodiment, R.sup.5 is C.sub.1-4alkyl. It is an
embodiment that R.sup.5 is methyl.
[0243] In an embodiment, R.sup.6 is selected from the group
consisting of C.sub.1-4alkyl, OH, C.sub.1-4alkoxy,
C.sub.6-10aryloxy, --NH--C.sub.1-4alkyl, --N(C.sub.1-4alkyl).sub.2,
C.sub.1-4alkylene-N(C.sub.1-4alkyl).sub.2,
C.sub.1-4alkylene-NH--C.sub.1-4alkyl, C.sub.3-8cycloalkyl,
heterocycloalkyl, C.sub.6-10aryl and a 5- or 6-membered heteroaryl.
In another embodiment, R.sup.6 is selected from the group
consisting of C.sub.1-4alkyl, OH, C.sub.1-4alkoxy,
--NH--C.sub.1-4alkyl and --N(C.sub.1-4alkyl).sub.2.
[0244] In an embodiment, R.sup.7 and R.sup.8 are each independently
selected from the group consisting of H, C.sub.1-4alkyl,
C.sub.1-4haloalkyl, C.sub.2-4alkenyl, C.sub.2-4alkynyl,
C.sub.3-8cycloalkyl, C.sub.1-4alkylene-C.sub.3-8cycloalkyl,
heterocycloalkyl, C.sub.6-10aryl, C.sub.1-4alkylene-C.sub.6-10aryl,
C.sub.1-4alkylene-heterocycloalkyl, heteroaryl, and
C.sub.1-4alkylene-heteroaryl, wherein any cyclic moiety is
optionally fused to a further 5- to 7-membered
heterocycloalkyl.
[0245] In an embodiment, the compounds of Formula I have the
following relative stereochemistry:
##STR00007##
[0246] In an embodiment, the compound of the present application is
selected from the compounds of Examples 1 to 33 as illustrated
below or a pharmaceutically acceptable salt, solvate or prodrug
thereof: [0247] 2-Methyl-thiazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
[0248] 2-Methyl-thiazole-5-carboxylic
acid-((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarb-
amoyl]-2-methoxy-ethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
[0249] 2-Methyl-thiazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-methoxyethyl)-amide; [0250]
2-Methyl-oxazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
[0251] 3-Methyl-isoxazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
[0252] Thiazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxoethylcarbamoyl]-
-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)amide;
[0253] Oxazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
[0254] 5-Methyl-thiophene-2-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
[0255] 5-Methyl-furan-2-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
[0256] Thiophene-2-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
[0257]
N-[(1S)-2-[[(1S)-2-[[(1S)-1-benzyl-2-[(2R)-2-methyloxiran-2-yl]-2-oxo-eth-
yl]amino]-1
(difluoromethoxymethyl)-2-oxo-ethyl]amino]-1-(difluoromethoxymethyl)-2-ox-
o-ethyl]-1H-1,2,4-triazole-5-carboxamide; [0258]
N-[(1S)-1-(difluoromethoxymethyl)-2-[[(1S)-1-(difluoromethoxymethyl)-2-[[-
(1S)-3-methyl-1-[(2R)-2-methyloxirane-2-carbonyl]butyl]-amino]-2-oxo-ethyl-
]amino]-2-oxo-ethyl]-2-methyl-thiazole-5-carboxamide; [0259]
Pyridine-2-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
[0260]
N--((S)-1-{(S)-1-[(S)-1-Benzyl-2-((R)-2-methyl-oxiranyl)-2-oxo-ethylcarba-
moyl]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-nicotinamid-
e; [0261] Pyridine-2-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
[0262]
N-[(1S)-2-[[(1S)-2-[[(1S)-1-benzyl-2-[(2R)-2-methyloxiran-2-yl]-2-oxo-eth-
yl]amino]-1-(difluoromethoxymethyl)-2-oxo-ethyl]amino]-1-(difluoromethoxym-
ethyl)-2-oxo-ethyl]-pyrimidine-2-carboxamide; [0263]
[1,2,4]Triazine-3-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
[0264] Pyrimidine-4-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
[0265] 2-Methyl-thiazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-3-methylbutylcarbamoyl}-2-difluoromethoxyethyl)-amide; [0266]
2-Methyl-thiazole-5-carboxylic acid
((S)-2-difluoromethoxy-1-{(S)-3-methyl-1-[(S)-3-methyl-1-((R)-2-methyloxi-
ranecarbonyl)-butylcarbamoyl]-butylcarbamoyl}-ethyl)-amide, [0267]
2-Methyl-thiazole-5-carboxylic acid
((S)-1-{(S)-2-difluoromethoxy-1-[(S)-3-methyl-1-((R)-2-methyloxiranecarbo-
nyl)-butylcarbamoyl]-ethylcarbamoyl}-3-methylbutyl)-amide; [0268]
2-Methyl-thiazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-3-methylbutyl)-amide; [0269]
2-Methyl-thiazole-5-carboxylic acid
((S)-1-{(S)-2-difluoromethoxy-1-[(S)-3-methyl-1-((R)-2-methyloxiranecarbo-
nyl)-butylcarbamoyl]-ethylcarbamoyl}-2-phenylethyl)-amide; [0270]
2-Methyl-thiazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyl-oxiranyl)-2-oxo-ethylcarbamoy-
l]-2-difluoromethoxyethylcarbamoyl}-2-phenylethyl)-amide; [0271]
2S)-2-[[(2S)-3-(difluoromethoxy)-2-[(2-morpholinoacetyl)amino]propanoyl]a-
mino]-N-[(1S)-3-methyl-1-[(2R)-2-methyloxirane-2-carbonyl]butyl]-3-phenylp-
ropanamide; [0272]
(S)--N--{(S)-1-[(S)-1-Benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethyl-carbamo-
yl]-2-difluoromethoxyethyl}-3-difluoromethoxy-2-(2-morpholin-4-yl-acetylam-
ino)-propionamide; [0273]
(2S)--N-[(1S)-1-benzyl-2-[(2R)-2-methyloxiran-2-yl]-2-oxo-ethyl]-2-[[(2S)-
-3-(difluoromethoxy)-2-[(2-morpholinoacetyl)amino]propanoyl]amino]-4-methy-
lpentanamide; [0274]
(2S)-2-[[(2S)-3-(difluoromethoxy)-2-[(2-morpholinoacetyl)amino]propanoyl]-
amino]-4-methyl-N-[(1S)-3-methyl-1-[(2R)-2-methyloxirane-2-carbonyl]butyl]-
pentanamide; [0275]
(2S)--N-[(1S)-1-benzyl-2-[(2R)-2-methyloxiran-2-yl]-2-oxo-ethyl]-2-[[(2S)-
-3-(difluoromethoxy)-2-[(2-morpholinoacetyl)amino]propanoyl]amino]-3-(1-me-
thylcyclohexa-1,3,5-trien-1-yl)propanamide; [0276]
(S)-4-Methyl-2-(2-morpholin-4-yl-acetylamino)-pentanoic acid
{(S)-2-difluoromethoxy-1-[(S)-3-methyl-1-((R)-2-methyloxiranecarbonyl)-bu-
tylcarbamoyl]-ethyl}-amide; [0277]
(S)-4-Methyl-2-(2-morpholin-4-yl-acetylamino)-pentanoic acid
{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl]-2-dif-
luoromethoxyethyl}-amide; [0278]
(2S)--N-[(1S)-2-[[(1S)-1-benzyl-2-[(2R)-2-methyloxiran-2-yl]-2-oxo-ethyl]-
amino]-1(difluoromethoxymethyl)-2-oxo-ethyl]-2-[(2-morpholinoacetyl)amino]-
-3-phenylpropanamide; and [0279]
(2S)-3-(difluoromethoxy)-N-[(1S)-1-(difluoromethoxymethyl)-2-[[(1S)-3-met-
hyl-1-[(2R)-2-methyloxirane-2-carbonyl]butyl]amino]-2-oxo-ethyl]-2-[(2-mor-
pholinoacetyl)amino]-propanamide.
[0280] In another embodiment, the compound of the present
application is selected from: [0281] 2-Methyl-thiazole-5-carboxylic
acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide;
[0282]
(2S)-2-[[(2S)-3-(difluoromethoxy)-2-[(2-morpholinoacetyl)amino]propanoyl]-
amino]-N-[(1S)-3-methyl-1-[(2R)-2-methyloxirane-2-carbonyl]butyl]-3-phenyl-
-propanamide; [0283]
(S)--N--{(S)-1-[(S)-1-benzyl-2-((R)-2-methyl-oxiranyl)-2-oxo-ethylcarbamo-
yl]-2-difluoromethoxyethyl}-3-difluoromethoxy-2-(2-morpholin-4-yl-acetylam-
ino)-propionamide; [0284]
(2S)--N-[(1S)-1-benzyl-2-[(2R)-2-methyloxiran-2-yl]-2-oxo-ethyl]-2-[[(2S)-
-3-(difluoromethoxy)-2-[(2-morpholinoacetyl)amino]propanoyl]amino]-4-methy-
lpentanamide; [0285]
(2S)-2-[[(2S)-3-(difluoromethoxy)-2-[(2-morpholino-acetyl)amino]propanoyl-
]amino]-4-methyl-N-[(1S)-3-methyl-1-[(2R)-2-methyloxirane-2-carbonyl]butyl-
]pentanamide; [0286]
(2S)--N-[(1S)-1-benzyl-2-[(2R)-2-methyloxiran-2-yl]-2-oxo-ethyl]-2-[[(2S)-
-3-(difluoromethoxy)-2-[(2-morpholinoacetyl)amino]propanoyl]amino]-3-(1-me-
thylcyclohexa-1,3,5-trien-1-yl)propanamide; and [0287]
(2S)-3-(difluoromethoxy)-N-[(1S)-1-(difluoromethoxymethyl)-2-[[(1S)-3-met-
hyl-1-[(2R)-2-methyloxirane-2-carbonyl]butyl]amino]-2-oxo-ethyl]-2-[(2-mor-
pholinoacetyl)amino]propanamide.
[0288] In another embodiment, the compound of the present
application is 2-methyl-thiazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide.
[0289] In an embodiment, the present application also includes a
compound of Formula (I):
##STR00008##
wherein:
[0290] R.sup.1 is selected from the group consisting of
C.sub.1-6-alkyl, C.sub.1-6-alkyloxy, C.sub.1-6-alkyloxyoalkyl,
C.sub.1-6-alkenyloxyhaloalkyl, C.sub.1-6-alkynyloxyhaloalkyl,
C.sub.1-6-alkylhalo, C.sub.2-6-alkenyl, C.sub.2-6-alkynyl,
C.sub.3-8-cycloalkyl, C.sub.1-6-alkyl-C.sub.3-8-cycloalkyl, aryl,
heteroaryl, C.sub.1-6-alkylaryl, C.sub.1-6-alkylheteroaryl,
C.sub.1-6-alkylheterocycloalkyl, C(O)H, (CO)R.sup.7, O(CO)R.sup.7,
C(O)OR.sup.7, C.sub.1-6-alkylOR.sup.7, C.sub.1-6-alkyl(CO)R.sup.7,
C.sub.0-6-alkylCO.sub.2R.sup.7, C.sub.1-6-alkylcyano,
C.sub.1-6-alkylNR.sup.7R.sup.8, C.sub.1-6-alkyl(CO)NR.sup.7R.sup.8,
C.sub.1-6-alkylNR.sup.6(CO)R.sup.8,
C.sub.1-6-alkylNR.sup.7(CO)NR.sup.7R.sup.8,
C.sub.1-6-alkylSR.sup.7, C.sub.1-6-alkyl(SO)R.sup.7,
C.sub.1-6-alkylSO.sub.2R.sup.6,
C.sub.1-6-alkyl(SO.sub.2)NR.sup.7R.sup.8,
C.sub.1-6-alkylNR.sup.7(SO.sub.2)R.sup.8,
C.sub.1-6-alkylNR.sup.7(SO.sub.2)NR.sup.7R.sup.8,
(CO)NR.sup.7R.sup.8, C.sub.1-6-alkylNR.sup.7(CO)OR.sup.8, and a 3-
to 7-membered ring that may contain one or more heteroatoms
independently selected from the group consisting of N, O and S,
wherein any cyclic moiety is optionally fused to a 5- to 7-membered
ring that may contain one or more heteroatoms independently
selected from the group consisting of N, O and S;
[0291] X is selected from the group consisting of hydrogen, carbon,
oxygen, nitrogen, sulfur, C.sub.1-6-alkyl, C.sub.1-6-alkyloxy,
C.sub.1-6-alkyloxyoalkyl, C.sub.1-6-alkenyloxyamminoalkyl,
C.sub.1-6-alkynyloxyhaloalkyl, C.sub.1-6-alkylhalo,
C.sub.2-6-alkenyl, C.sub.2-6-alkynyl, C.sub.3-8-cycloalkyl,
C.sub.1-6-alkyl-C.sub.3-8-cycloalkyl, aryl, heteroaryl,
C.sub.1-6-alkylaryl, C.sub.1-6-alkylheteroaryl,
C.sub.1-6-alkylheterocycloalkyl;
[0292] R.sup.2, R.sup.3 and R.sup.4, are selected from the group
consisting of C.sub.1-6-alkyl, C.sub.1-6-alkyloxy,
C.sub.1-6-alkyloxyoalkyl, C.sub.1-6-alkenyloxyhaloalkyl,
C.sub.1-6-alkynyloxyhaloalkyl, C.sub.1-6-alkylhalo,
C.sub.2-6-alkenyl, C.sub.2-6-alkynyl, C.sub.3-8-cycloalkyl,
C.sub.1-6-alkyl-C.sub.3-8-cycloalkyl, aryl, heteroaryl,
C.sub.1-6-alkylaryl, C.sub.1-6-alkylheteroaryl,
C.sub.1-6-alkylheterocycloalkyl, C(O)H, (CO)R.sup.7, O(CO)R.sup.7,
C(O)OR.sup.7, C.sub.1-6-alkylOR.sup.7, C.sub.1-6-alkyl(CO)R.sup.7,
C.sub.0-6-alkylCO.sub.2R.sup.7, C.sub.1-6-alkylcyano,
C.sub.1-6-alkylNR.sup.7R.sup.8, C.sub.1-6-alkyl(CO)NR.sup.7R.sup.8,
C.sub.1-6-alkylNR.sup.6(CO)R.sup.8,
C.sub.1-6-alkylN.sup.7(CO)NR.sup.7R.sup.8, C.sub.1-6-alkylSR.sup.7,
C.sub.1-6-alkyl(SO)R.sup.7, C.sub.1-6-alkylSO.sub.2R.sup.6,
C.sub.1-6-alkyl(SO.sub.2)NR.sup.7R.sup.8,
C.sub.1-6-alkylNR.sup.7(SO.sub.2)R.sup.8,
C.sub.1-6-alkylNR.sup.7(SO.sub.2)NR.sub.7R.sup.8,
(CO)NR.sup.7R.sup.8, C.sub.1-6-alkylNR.sup.7(CO)OR.sup.8, and a 3-
to 7-membered ring that may contain one or more heteroatoms
independently selected from the group consisting of N, O and S,
wherein any cyclic moiety is optionally fused to a 5- to 7-membered
ring that may contain one or more heteroatoms independently
selected from the group consisting of N, O and S; with the proviso
that at least R.sup.2, R.sup.3 or R.sup.4 is
C.sub.1-6-alkyloxyhaloalkyl, optionally substituted with one or
more independently-selected groups R.sup.6; and cannot
simultaneously be C.sub.1-6-alkyloxyoalkyl;
[0293] R.sup.5, is selected from the group consisting of H,
C.sub.1-6-alkyl, C.sub.1-6-alkylhalo, C.sub.2-6-alkenyl,
C.sub.2-6-alkynyl, C.sub.3-8-cycloalkyl,
C.sub.1-6-alkyl-C.sub.3-8-cycloalkyl, aryl, heteroaryl, and a 3- to
7-membered ring that may contain one or more heteroatoms
independently selected from the group consisting of N, O and S;
[0294] R.sup.6 is selected from the group consisting of H,
C.sub.1-6-alkyl, F, Cl, Br, I, OH, --O--(CH.sub.2).sub.2,3--O--,
OC.sub.1-6-alkyl, OC.sub.1-6-aryl, NH--C.sub.1-6-alkyl, and
N(C.sub.1-6-alkyl).sub.2, C.sub.1-6-alkyl-N(C.sub.1-6-alkyl).sub.2,
C.sub.1-6-alkyl-NH--C.sub.1-6-alkyl cycloalkyl, heterocycloalkyl,
aryl and heteroaryl;
[0295] R.sup.7 and R.sup.8 are independently selected from the
group consisting of H, C.sub.1-6-alkyl, C.sub.1-6-alkylhalo,
C.sub.2-6-alkenyl, C.sub.2-6-alkynyl, C.sub.3-8-cycloalkyl,
C.sub.1-6-alkyl-C.sub.3-8-cycloalkyl, cycloalkyl, aryl,
C.sub.1-6-alkylaryl, C.sub.0-6-alkyl-heterocycloalkyl, heteroaryl,
and C.sub.1-6alkylheteroaryl, wherein any cyclic moiety is
optionally fused to a 5- to 7-membered ring that may contain one or
more heteroatoms independently selected from the group consisting
of C, N, O and S.
[0296] In an embodiment, X is C.sub.0-7 alkyl and R.sup.1 is a 5-
or -6 membered heteroaryl and acyclic and 3-7-membered amine.
[0297] In another embodiment, X is C.sub.0-7 alkyl, R.sup.1 is
selected from morpholine, 1,4-oxazepane, thiomorpholine,
1,4-thiazepane, 1,4-thiazepane 1-oxide, 1,4-thiazepane 1,1-dioxide,
1,4-thiazinane 1-oxide, 1,4-thiazinane 1,1-dioxide, aziridine,
azetidine, pyrrolidine, piperazine and 1,4-diazepane, oxadiazole
isoxazole, isothiazole, furan, thiophene, oxazole, thiazole,
pyrazole, and imidazole.
[0298] In another embodiment, at least one of R.sup.2, R.sup.3 or
R.sup.4 is selected from C.sub.1-6 alkoxyhaloalkyl, in a further
certain aspect, at least one of R.sup.2, R.sup.3 and R.sup.4 or all
is selected from mono-fluoromethyl, di-fluoromethyl and
tri-fluoromethyl moiety groups.
[0299] In another embodiment, at least one of R.sup.2 and R.sup.3
is selected from C.sub.1-6-alkoxyhaloalkyl, in a further certain
aspect, at least one of R.sup.2 and R.sup.3 or both are selected
from di-fluoromethyl, moiety.
[0300] In another embodiment, R.sup.4 is selected from C.sub.1-6
alkyl, C.sub.1-6 alkyl, C.sub.1-6 alkylcycloalkyl and C.sub.1-6
alkylaryl. In certain such embodiment, R.sup.4 is selected from
phenylmethyl, phenyl-ethyl, 2-methyl-butanyl, 2,2-dimethyl-butanyl,
preferably phenyl-methyl and 2-Methyl-butanyl.
III. Preparation of Compounds of the Application
[0301] Compounds of the present application can be prepared by
various synthetic processes. The choice of particular structural
features and/or substituents may influence the selection of one
process over another. The selection of a particular process to
prepare a given compound of Formula I is within the purview of the
person of skill in the art. Some starting materials for preparing
compounds of the present application are available from commercial
chemical sources. Other starting materials, for example as
described below, are readily prepared from available precursors
using straightforward transformations that are well known in the
art.
[0302] In an embodiment, the compounds of Formula I are generally
prepared according to the process illustrated in Scheme I.
Variables in the following schemes are as defined above for Formula
I unless otherwise specified.
##STR00009##
[0303] In an embodiment, as shown in Scheme 1, the compounds of the
present application are prepared by coupling of a dipeptide of
Formula II with an epoxyketone of Formula III via the formation of
a peptide bond. Methods for coupling compounds through peptide
(amide) bonds are well known in the art and described, for example,
in The Peptides: Analysis, Synthesis, Biology, Vol. I., eds.
Academic Press, 1979.
[0304] In an embodiment, the intermediate compound of Formula II is
prepared according to standard procedures for peptide bond
formation as illustrated in Scheme II, wherein the compounds of
Formula IV and V are coupled via amide bond formation. For example,
the compounds of Formula IV (wherein R.sup.1 is optionally a
protecting group, such as Boc or Cbz) and Formula V (wherein A is,
for example an alkyl or benzyl group) are obtained from commercial
sources or prepared by methods known in the art. Examples of the
dipeptide of Formula II are provided in specific examples described
below.
##STR00010##
[0305] In an embodiment, epoxyketones of Formula III are prepared
as illustrated in Scheme III using modified literature methods
[see, for example, Bioorg. Med. Chem. Lett. 2007, 17, 6169-6171;
Bioorg. Med. Chem. Lett. 1999, 9, 2283-2288; Eur. J. Org. Chem.
2005, 4829-4834; and J. Med. Chem. 2009, 52, 3028-3038].
##STR00011##
[0306] As shown in Scheme III, epoxyketone compounds of Formula
III, in one embodiment, are prepared from a protected amino acid of
Formula VI which is transformed to the corresponding Weinreb amide
of Formula VII [see, for example, Synthesis 1983, 676; Bioorg. Med.
Chem. Lett. 1999, 2283-2288], followed by an appropriate lithium,
zinc or Grignard reagent condensation leading to the unsaturated
ketone of Formula VIII. Subsequent epoxidation with alkaline
hydrogen peroxide provides epoxide derivatives of Formula IX as a
mixture of diastereomers, which are readily separated by column
chromatography. Removal of the protecting group (PG) by a suitable
method such as a hydrogenolysis reaction (for example, wherein PG
is a Cbz group) or hydrolysis in acidic conditions (for example,
wherein PG is a Boc protecting group), gives the epoxides of
Formula III in salt form, such as a salt of TFA.
[0307] In an alternative embodiment, according to Scheme IV,
epoxyketone intermediates of Formula III are prepared from an
unsaturated ketone of Formula VIII, which is reduced to the
corresponding allylic alcohol of Formula X. Subsequently, Sharpless
asymmetric epoxidation leads to hydroxyl-ketone compounds of
Formula XI which are oxidized to provide epoxyketone intermediates
of Formula IX, which are subsequently hydrolyzed to provide
intermediate compounds of Formula III [see, for example, J. Med.
Chem. 2009, 52, 3028-3038; Tetrahedron: Asymmetry 2001, 12,
943-947].
##STR00012##
[0308] In an embodiment, as a representative example of the
incorporation of a C.sub.1-6alkylene-O--C.sub.1-6haloalkyl group in
R.sup.2, R.sup.3 and/or R.sup.4 of the compounds of the
application, a precursor compound to the compounds of Formula I,
for example a compound of the Formula II, III, IV or V, or
protected forms thereof, wherein R.sup.2, R.sup.3 and/or R.sup.4 is
C.sub.1-6alkylene-OH is reacted with, for example,
2-fluorosulfonyldifluoroacetic acid in the presence of a metal
catalyst, such as copper (I) iodide, under conditions to covert the
C.sub.1-6alkylene-OH to C.sub.1-6alkylene-OCHF.sub.2. A person
skilled in the art would know other methods of functionalizing the
C.sub.1-6alkylene-OH group with alternative C.sub.1-6haloalkyl
group using methods and reagents available in the art.
[0309] Another aspect of the present application provides processes
for preparing compounds of Formula I, or salts, solvates or
prodrugs thereof. Processes for the preparation of the compounds in
the present application are described herein.
[0310] Accordingly, the present application includes a method for
preparing a compound of Formula I comprising reacting a compound of
a Formula II with a compound of a Formula III:
##STR00013##
Wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4 and R.sup.5 are as
defined above for the compounds of Formula I, or are protected
forms thereof, under conditions for the formation of an amide bond;
and removal of protecting groups if present.
[0311] In another embodiment, the compounds of Formula II are
prepared by reacting a compound of the Formula VIII:
##STR00014##
wherein PG is a protecting group and R.sup.4 and R.sup.5 are as
defined above for the compounds of Formula I, or are protected
forms thereof, under conditions for the epoxidation of the double
bond followed by removal of protecting groups.
[0312] In an alternative embodiment, the compounds of Formula II
are prepared by reacting a compound of the Formula VIII:
##STR00015##
wherein PG is a protecting group and R.sup.4 and R.sup.5 are as
defined above for the compounds of Formula I, or are protected
forms thereof, under conditions for the reduction of the ketone to
the corresponding alcohol, followed by epoxidation of the double
bond, followed by oxidation of the alcohol and then removal of
protecting groups.
[0313] In addition, it is intended that the present application
cover compounds made either using standard organic synthetic
techniques, including combinatorial chemistry or by biological
methods, such as bacterial digestion, metabolism, enzymatic
conversion, and the like.
[0314] Throughout the processes described herein it is to be
understood that, where appropriate, suitable protecting groups will
be added to, and subsequently removed from, the various reactants
and intermediates in a manner that will be readily understood by
one skilled in the art. Conventional procedures for using such
protecting groups as well as examples of suitable protecting groups
are described, for example, in "Protective Groups in Organic
Synthesis", T. W. Green, P. G. M. Wuts, Wiley-lnterscience, New
York, (1999). It is also to be understood that a transformation of
a group or substituent into another group or substituent by
chemical manipulation can be conducted on any intermediate or final
product on the synthetic path toward the final product, in which
the possible type of transformation is limited only by inherent
incompatibility of other functionalities carried by the molecule at
that stage to the conditions or reagents employed in the
transformation. Such inherent incompatibilities, and ways to
circumvent them by carrying out appropriate transformations and
synthetic steps in a suitable order, will be readily understood to
one skilled in the art. Examples of transformations are given
herein, and it is to be understood that the described
transformations are not limited only to the generic groups or
substituents for which the transformations are exemplified.
References and descriptions of other suitable transformations are
given in "Comprehensive Organic Transformations--A Guide to
Functional Group Preparations" R. C. Larock, VHC Publishers, Inc.
(1989). References and descriptions of other suitable reactions are
described in textbooks of organic chemistry, for example, "Advanced
Organic Chemistry", March, 4th ed. McGraw Hill (1992) or, "Organic
Synthesis", Smith, McGraw Hill, (1994). Techniques for purification
of intermediates and final products include, for example, straight
and reversed phase chromatography on column or rotating plate,
recrystallisation, distillation and liquid-liquid or solid-liquid
extraction, which will be readily understood by one skilled in the
art.
IV. Compositions
[0315] The compounds of the present application are suitably
formulated in a conventional manner into compositions using one or
more carriers. Accordingly, the present application also includes a
composition comprising one or more compounds of the application and
a carrier. In another embodiment, the compounds of the application
are suitably formulated into pharmaceutical compositions for
administration to subjects in a biologically compatible form
suitable for administration in vivo. Accordingly, the present
application further includes a pharmaceutical composition
comprising one or more compounds of the application and a
pharmaceutically acceptable carrier.
[0316] The compounds of the application are administered to a
subject in a variety of forms depending on the selected route of
administration, as will be understood by those skilled in the art.
A compound of the application is administered, for example, by
oral, parenteral, buccal, sublingual, nasal, rectal, patch, pump or
transdermal administration and the pharmaceutical compositions
formulated accordingly. In an embodiment, administration is by
means of a pump for periodic or continuous delivery.
[0317] Parenteral administration includes intravenous,
intra-arterial, intraperitoneal, subcutaneous, intramuscular,
transepithelial, nasal, intrapulmonary (for example, by use of an
aerosol), intrathecal, rectal and topical (including the use of a
patch or other transdermal delivery device) modes of
administration. In an embodiment, parenteral administration is by
continuous infusion over a selected period of time. Conventional
procedures and ingredients for the selection and preparation of
suitable compositions are described, for example, in Remington's
Pharmaceutical Sciences (2000-20th edition) and in The United
States Pharmacopeia: The National Formulary (USP 24 NF19) published
in 1999.
[0318] In an embodiment, a compound of the application is orally
administered, for example, with an inert diluent or with an
assimilable edible carrier, or it is enclosed in hard or soft shell
gelatin capsules, or it may be compressed into tablets, or it is
incorporated directly with the food of the diet. In a further
embodiment, for oral therapeutic administration, the compound is
incorporated with excipient and used in the form of ingestible
tablets, buccal tablets, troches, capsules, caplets, pellets,
granules, lozenges, chewing gum, powders, syrups, elixirs, wafers,
aqueous solutions or suspensions, and the like. In the case of
tablets, carriers that are used include lactose, corn starch,
sodium citrate and salts of phosphoric acid. Pharmaceutically
acceptable excipients include binding agents (e.g., pregelatinized
maize starch, polyvinylpyrrolidone or hydroxypropyl
methylcellulose); fillers (e.g., lactose, microcrystalline
cellulose or calcium phosphate); lubricants (e.g., magnesium
stearate, talc or silica); disintegrants (e.g., potato starch or
sodium starch glycolate); or wetting agents (e.g., sodium lauryl
sulphate). In an embodiment, the tablets are coated by methods well
known in the art. In the case of tablets, capsules, caplets,
pellets or granules for oral administration, pH sensitive enteric
coatings, such as Eudragits.TM., designed to control the release of
active ingredients are optionally used. Oral dosage forms also
include modified release, for example immediate release and
timed-release, formulations. Examples of modified-release
formulations include, for example, sustained-release (SR),
extended-release (ER, XR, or XL), time-release or timed-release,
controlled-release (CR), or continuous-release (CR or Contin),
employed, for example, in the form of a coated tablet, an osmotic
delivery device, a coated capsule, a microencapsulated microsphere,
an agglomerated particle, e.g., as of molecular sieving type
particles, or, a fine hollow permeable fiber bundle, or chopped
hollow permeable fibers, agglomerated or held in a fibrous packet.
Timed-release compositions are formulated, for e.g. in liposomes or
those wherein the active compound is protected with differentially
degradable coatings, such as by microencapsulation, multiple
coatings, etc. Liposome delivery systems include, for example,
small unilamellar vesicles, large unilamellar vesicles and
multilamellar vesicles. In an embodiment, liposomes are formed from
a variety of phospholipids, such as cholesterol, stearylamine or
phosphatidylcholines. For oral administration in a capsule form,
useful carriers or diluents include lactose and dried corn
starch.
[0319] Liquid preparations for oral administration take the form
of, for example, solutions, syrups or suspensions, or they are
suitably presented as a dry product for constitution with water or
other suitable vehicle before use. When aqueous suspensions and/or
emulsions are administered orally, the compound of the application
is suitably suspended or dissolved in an oily phase that is
combined with emulsifying and/or suspending agents. If desired,
certain sweetening and/or flavoring and/or coloring agents are
added. Such liquid preparations for oral administration are
prepared by conventional means with pharmaceutically acceptable
additives such as suspending agents (e.g., sorbitol syrup, methyl
cellulose or hydrogenated edible fats); emulsifying agents (e.g.,
lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily
esters or ethyl alcohol); and/or preservatives (e.g., methyl or
propyl p-hydroxybenzoates or sorbic acid). Useful diluents include
lactose and/or high molecular weight polyethylene glycols.
[0320] It is also possible to freeze-dry the compounds of the
application and use the lyophilizates obtained, for example, for
the preparation of products for injection.
[0321] In a further embodiment, a compound of the application is
administered parenterally. For example, solutions of a compound of
the application are prepared in water suitably mixed with a
surfactant such as hydroxypropylcellulose. Dispersions are also
prepared in glycerol, liquid polyethylene glycols, DMSO and
mixtures thereof with or without alcohol, and in oils. Under
ordinary conditions of storage and use, these preparations contain
a preservative to prevent the growth of microorganisms. A person
skilled in the art would know how to prepare suitable formulations.
For parenteral administration, sterile solutions of the compounds
of the application are usually prepared, and the pH's of the
solutions are suitably adjusted and buffered. For intravenous use,
the total concentration of solutes should be controlled to render
the preparation isotonic. For ocular administration, ointments or
droppable liquids are, for example, delivered by ocular delivery
systems known to the art such as applicators or eye droppers. In an
embodiment, such compositions include mucomimetics such as
hyaluronic acid, chondroitin sulfate, hydroxypropyl methylcellulose
and/or polyvinyl alcohol, preservatives such as sorbic acid, EDTA
and/or benzyl chromium chloride, and the usual quantities of
diluents or carriers. For pulmonary administration, diluents and/or
carriers will be selected to be appropriate to allow the formation
of an aerosol.
[0322] In a further embodiment, the compounds of the application
are formulated for parenteral administration by injection,
including using conventional catheterization techniques or
infusion. Formulations for injection are, for example, presented in
unit dosage form, e.g., in ampoules or in multi-dose containers,
with an added preservative. In an embodiment, the compositions take
such forms as sterile suspensions, solutions or emulsions in oily
or aqueous vehicles, and, optionally contain formulating agents
such as suspending, stabilizing and/or dispersing agents. In all
cases, the form must be sterile and must be fluid to the extent
that easy syringability exists. Alternatively, the compounds of the
application are suitably in a sterile powder form for
reconstitution with a suitable vehicle, e.g., sterile pyrogen-free
water, before use.
[0323] In an embodiment, compositions for nasal administration are
conveniently formulated as aerosols, drops, gels and powders. For
intranasal administration or administration by inhalation, the
compounds of the application are conveniently delivered in the form
of a solution, dry powder formulation or suspension from a pump
spray container that is squeezed or pumped by the patient or as an
aerosol spray presentation from a pressurized container or a
nebulizer. Aerosol formulations typically comprise a solution or
fine suspension of the active substance in a physiologically
acceptable aqueous or non-aqueous solvent and are usually presented
in single or multidose quantities in sterile form in a sealed
container, which can take the form of a cartridge or refill for use
with an atomising device. Alternatively, the sealed container is a
unitary dispensing device such as a single dose nasal inhaler or an
aerosol dispenser fitted with a metering valve which is intended
for disposal after use. Where the dosage form comprises an aerosol
dispenser, it will contain a propellant which is, for example, a
compressed gas such as compressed air or an organic propellant such
as fluorochlorohydrocarbon. Suitable propellants include but are
not limited to dichlorodifluoromethane, trichlorofluoromethane,
dichlorotetrafluoroethane, heptafluoroalkanes, carbon dioxide or
another suitable gas. In the case of a pressurized aerosol, the
dosage unit is suitably determined by providing a valve to deliver
a metered amount. In an embodiment, the pressurized container or
nebulizer contains a solution or suspension of the active compound.
Capsules and cartridges (made, for example, from gelatin) for use
in an inhaler or insufflator are, for example, formulated
containing a powder mix of a compound of the application and a
suitable powder base such as lactose or starch. In an embodiment,
the aerosol dosage forms also take the form of a pump-atomizer.
[0324] Compositions suitable for buccal or sublingual
administration include tablets, lozenges, and pastilles, wherein
the active ingredient is formulated with a carrier such as sugar,
acacia, tragacanth, and/or gelatin and glycerine. Compositions for
rectal administration are conveniently in the form of suppositories
containing a conventional suppository base such as cocoa
butter.
[0325] Suppository forms of the compounds of the application are
useful for vaginal, urethral and rectal administrations. Such
suppositories will generally be constructed of a mixture of
substances that is solid at room temperature but melts at body
temperature. The substances commonly used to create such vehicles
include but are not limited to theobroma oil (also known as cocoa
butter), glycerinated gelatin, other glycerides, hydrogenated
vegetable oils, mixtures of polyethylene glycols of various
molecular weights and/or fatty acid esters of polyethylene glycol.
See, for example: Remington's Pharmaceutical Sciences, 16th Ed.,
Mack Publishing, Easton, Pa., 1980, pp. 1530-1533 for further
discussion of suppository dosage forms.
[0326] In an embodiment, compounds of the application are coupled
with soluble polymers as targetable drug carriers. Such polymers
include, for example, polyvinylpyrrolidone, pyran copolymer,
polyhydroxypropylmethacrylamide-phenol,
polyhydroxy-ethylaspartamide-phenol, or
polyethyleneoxide-polylysine substituted with palmitoyl residues.
In a further embodiment, compounds of the application are coupled
to a class of biodegradable polymers useful in achieving controlled
release of a drug, for example, polylactic acid, polyglycolic acid,
copolymers of polylactic and polyglycolic acid, polyepsilon
caprolactone, polyhydroxy butyric acid, polyorthoesters,
polyacetals, polydihydropyrans, polycyanoacrylates and crosslinked
or amphipathic block copolymers of hydrogels.
[0327] The compounds of the application including pharmaceutically
acceptable salts, solvates and prodrugs thereof are suitably used
on their own but will generally be administered in the form of a
pharmaceutical composition in which the one or more compounds of
the application (the active ingredient) is in association with a
pharmaceutically acceptable carrier. Depending on the mode of
administration, the pharmaceutical composition will comprise from
about 0.05 wt % to about 99 wt % or about 0.10 wt % to about 70 wt
%, of the active ingredient, and from about 1 wt % to about 99.95
wt % or about 30 wt % to about 99.90 wt % of a pharmaceutically
acceptable carrier, all percentages by weight being based on the
total composition.
[0328] Compounds of the application are used alone or in
combination with other known agents useful for treating diseases,
disorders or conditions mediated by proteasome inhibition. When
used in combination with other agents useful in treating diseases,
disorders or conditions mediated by proteasome inhibition, it is an
embodiment that the compounds of the application are administered
contemporaneously with those agents. As used herein,
"contemporaneous administration" of two substances to a subject
means providing each of the two substances so that they are both
biologically active in the individual at the same time. The exact
details of the administration will depend on the pharmacokinetics
of the two substances in the presence of each other, and can
include administering the two substances within a few hours of each
other, or even administering one substance within 24 hours of
administration of the other, if the pharmacokinetics are suitable.
Design of suitable dosing regimens is routine for one skilled in
the art. In particular embodiments, two substances will be
administered substantially simultaneously, i.e., within minutes of
each other, or in a single composition that contains both
substances. It is a further embodiment of the present application
that a combination of agents is administered to a subject in a
non-contemporaneous fashion. In an embodiment, a compound of the
present application is administered with another therapeutic agent
simultaneously or sequentially in separate unit dosage forms or
together in a single unit dosage form. Accordingly, the present
application provides a single unit dosage form comprising a
compound of Formula I, an additional therapeutic agent, and a
pharmaceutically acceptable carrier.
[0329] The dosage of compounds of the application varies depending
on many factors such as the pharmacodynamic properties of the
compound, the mode of administration, the age, health and weight of
the recipient, the nature and extent of the symptoms, the frequency
of the treatment and the type of concurrent treatment, if any, and
the clearance rate of the compound in the subject to be treated.
One of skill in the art can determine the appropriate dosage based
on the above factors. In an embodiment, compounds of the
application are administered initially in a suitable dosage that is
optionally adjusted as required, depending on the clinical
response. Dosages will generally be selected to maintain a serum
level of compounds of the application from about 0.01 .mu.g/cc to
about 1000 .mu.g/cc, or about 0.1 .mu.g/cc to about 100 .mu.g/cc.
As a representative example, oral dosages of one or more compounds
of the application will range between about 1 mg per day to about
1000 mg per day for an adult, suitably about 1 mg per day to about
500 mg per day, more suitably about 1 mg per day to about 200 mg
per day. For parenteral administration, a representative amount is
from about 0.001 mg/kg to about 10 mg/kg, about 0.01 mg/kg to about
10 mg/kg, about 0.01 mg/kg to about 1 mg/kg or about 0.1 mg/kg to
about 1 mg/kg. For oral administration, a representative amount is
from about 0.001 mg/kg to about 10 mg/kg, about 0.1 mg/kg to about
10 mg/kg, about 0.01 mg/kg to about 1 mg/kg or about 0.1 mg/kg to
about 1 mg/kg. For administration in suppository form, a
representative amount is from about 0.1 mg/kg to about 10 mg/kg or
about 0.1 mg/kg to about 1 mg/kg. In an embodiment of the
application, compositions are formulated for oral administration
and the compounds are suitably in the form of tablets containing
0.25, 0.5, 0.75, 1.0, 5.0, 10.0, 20.0, 25.0, 30.0, 40.0, 50.0,
60.0, 70.0, 75.0, 80.0, 90.0, 100.0, 150, 200, 250, 300, 350, 400,
450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950 or 1000 mg of
active ingredient per tablet. In an embodiment, compounds of the
application are administered in a single daily dose or the total
daily dose is divided into two, three, four or more daily
doses.
V. Methods and Uses of the Application
[0330] The compounds of the application have been shown to be
capable of inhibiting proteasome activity.
[0331] Accordingly, the present application includes a method for
inhibiting proteasome in a cell, either in a biological sample or
in a patient, comprising administering an effective amount of one
or more compounds of the application to the cell. The application
also includes a use of one or more compounds of the application for
inhibition of proteasome in a cell as well as a use of one or more
compounds of the application for the preparation of a medicament
for inhibition of proteasome in a cell. The application further
includes one or more compounds of the application for use in
inhibiting proteasome in a cell.
[0332] As the compounds of the application have been shown to be
capable of inhibiting proteasome activity, the compounds of the
application are useful for treating diseases, disorders or
conditions mediated by proteasome inhibition. Therefore the
compounds of the present application are useful as medicaments.
Accordingly, the present application includes a compound of the
application for use as a medicament.
[0333] The present application also includes a method of treating a
disease, disorder or condition that is mediated by proteasome
inhibition comprising administering a therapeutically effective
amount of one or more compounds of the application to a subject in
need thereof.
[0334] The present application also includes a use of one or more
compounds of the application for treatment of a disease, disorder
or condition mediated by proteasome inhibition as well as a use of
one or more compounds of the application for the preparation of a
medicament for treatment of a disease, disorder or condition
mediated by proteasome inhibition. The application further includes
one or more compounds of the application for use in treating a
disease, disorder or condition mediated by proteasome
inhibition.
[0335] In an embodiment, the disease, disorder or condition
mediated by proteasome inhibition is a neoplastic disorder.
Accordingly, the present application also includes a method of
treating a neoplastic disorder comprising administering a
therapeutically effective amount of one or more compounds of the
application to a subject in need thereof. The present application
also includes a use of one or more compounds of the application for
treatment of a neoplastic disorder as well as a use of one or more
compounds of the application for the preparation of a medicament
for treatment of a neoplastic disorder. The application further
includes one or more compounds of the application for use in
treating a neoplastic disorder. In an embodiment, the treatment is
in an amount effective to ameliorate at least one symptom of the
neoplastic disorder, for example, reduced cell proliferation or
reduced tumor mass, among others, in a subject in need of such
treatment.
[0336] Compounds of the application have been demonstrated to be
effective against the cell lines of a 60 human tumor cell line
panel. Therefore in another embodiment of the present application,
the disease, disorder or condition mediated by proteasome
inhibition is cancer. Accordingly, the present application also
includes a method of treating cancer comprising administering a
therapeutically effective amount of one or more compounds of the
application to a subject in need thereof. The present application
also includes a use of one or more compounds of the application for
treatment of cancer as well as a use of one or more compounds of
the application for the preparation of a medicament for treatment
of cancer. The application further includes one or more compounds
of the application for use in treating cancer. In an embodiment,
the compound is administered for the prevention of cancer in a
subject such as a mammal having a predisposition for cancer.
[0337] In an embodiment, the cancer is selected from a cancer of
the skin, blood, prostate, colorectum, pancreas, kidney, ovary,
breast, for example mammary, liver, tongue and lung. In another
embodiment, the cancer is selected from leukaemia, lymphoma,
non-Hodgkin's lymphoma and multiple myeloma. In a further
embodiment of the present application, the cancer is selected from
leukemia, melanoma, lung cancer, colon cancer, brain cancer,
ovarian cancer, breast cancer, prostate cancer and kidney
cancer.
[0338] In an embodiment, the disease, disorder or condition
mediated by proteasome inhibition is a disease, disorder or
condition associated with an uncontrolled and/or abnormal cellular
activity affected directly or indirectly by proteasome inhibition.
In another embodiment, the uncontrolled and/or abnormal cellular
activity that is affected directly or indirectly by proteasome
inhibition is proliferative activity in a cell. Accordingly, the
application also includes a method of inhibiting proliferative
activity in a cell, comprising administering an effective amount of
one or more compounds of the application to the cell. The present
application also includes a use of one or more compounds of the
application for inhibition of proliferative activity in a cell as
well as a use of one or more compounds of the application for the
preparation of a medicament for inhibition of proliferative
activity in a cell. The application further includes one or more
compounds of the application for use in inhibiting proliferative
activity in a cell.
[0339] The present application also includes a method of inhibiting
uncontrolled and/or abnormal cellular activities affected directly
or indirectly by proteasome inhibition in a cell, either in a
biological sample or in a subject, comprising administering an
effective amount of one or more compounds of the application to the
cell. The application also includes a use of one or more compounds
of the application for inhibition of uncontrolled and/or abnormal
cellular activities affected directly or indirectly by proteasome
inhibition in a cell as well as a use of one or more compounds of
the application for the preparation of a medicament for inhibition
of uncontrolled and/or abnormal cellular activities affected
directly or indirectly by proteasome inhibition in a cell. The
application further includes one or more compounds of the
application for use in inhibiting uncontrolled and/or abnormal
cellular activities affected directly or indirectly by proteasome
inhibition in a cell.
[0340] The administration of the compound of Example 1 of the
present application in combination with either bortezomib or
dexamethasone has been demonstrated herein to be capable of being
synergistic. Accordingly, the present application also includes a
method of treating a disease, disorder or condition that is
mediated by proteasome inhibition comprising administering a
therapeutically effective amount of one or more compounds of the
application in combination with another known agent useful for
treatment of a disease, disorder or condition mediated by
proteasome inhibition to a subject in need thereof. The present
application also includes a use of one or more compounds of the
application in combination with another known agent useful for
treatment of a disease, disorder or condition mediated by
proteasome inhibition for treatment of a disease, disorder or
condition mediated by proteasome inhibition as well as a use of one
or more compounds of the application in combination with another
known agent useful for treatment of a disease, disorder or
condition mediated by proteasome inhibition for the preparation of
a medicament for treatment of a disease, disorder or condition
mediated by proteasome inhibition. The application further includes
one or more compounds of the application in combination with
another known agent useful for treatment of a disease, disorder or
condition mediated by proteasome inhibition for use in treating a
disease, disorder or condition mediated by proteasome inhibition.
In an embodiment, the disease, disorder or condition mediated by
proteasome inhibition is cancer such as multiple myeloma. In
another embodiment, the other known agent useful for treatment of a
disease, disorder or condition mediated by proteasome inhibition is
bortezomib or dexamethasone.
[0341] In a further embodiment, the disease, disorder or condition
mediated by proteasome inhibition is cancer and the one or more
compounds of the application are administered in combination with
one or more additional cancer treatments. In another embodiment,
the additional cancer treatment is selected from radiotherapy,
chemotherapy, targeted therapies such as antibody therapies and
small molecule therapies such as tyrosine-kinase inhibitors,
immunotherapy, hormonal therapy and anti-angiogenic therapies.
[0342] The present application also includes a method of inhibiting
the degradation of a protein by a proteasome capable of degrading
the protein, comprising contacting the proteasome with an effective
amount of one or more compounds of the application. The present
application further includes a use of one or more compounds of the
application for inhibition of the degradation of a protein by a
proteasome capable of degrading the protein as well as a use of one
or more compounds of the application for preparation of a
medicament for inhibition of the degradation of a protein by a
proteasome capable of degrading the protein. The present
application also includes one or more compounds of the application
for inhibiting the degradation of a protein by a proteasome capable
of degrading the protein.
[0343] In an embodiment, the protein is marked with ubiquitin. In
another embodiment, the protein is p53.
[0344] The present application also includes a method of treating
accelerated and/or enhanced proteolysis, comprising administering a
therapeutically effective amount of one or more compounds of the
application to a subject in need thereof. The present application
further includes a use of one or more compounds of the application
for treatment of accelerated and/or enhanced proteolysis as well as
a use of one or more compounds of the application for the
preparation of a medicament for treatment of accelerated and/or
enhanced proteolysis. The present application also includes one or
more compounds of the application for treating accelerated and/or
enhanced proteolysis. In an embodiment, the subject is a mammal
having or predisposed to accelerated and/or enhanced
proteolysis.
[0345] In another embodiment of the present application, the
disease, disorder or condition mediated by proteasome inhibition is
selected from a disease, disorder or condition associated with the
cell cycle, Endoplasmic Reticulum Associated Protein Degradation,
transcription factor regulation, gene expression, cell
differentiation, the immune response, angiogenesis and the
regulation or induction of apoptosis.
[0346] In another embodiment of the present application, the
disease, disorder or condition mediated by proteasome inhibition is
selected from viral infection, an inflammatory disease, an
autoimmune disease, heart disease, an age-related eye disease and a
neurodegenerative disease
[0347] In another embodiment of the present application, the
disease, disorder or condition mediated by proteasome inhibition is
selected from HIV infection, type-1 diabetes, type-2 diabetes,
allergic reactions, asthma, inflammatory arthritis, rheumatoid
arthritis, osteoporosis, osteoarthritis, psoriasis, seronegative
spondyloarthopathies, ankylosing spondylitis, systemic lupus
erythematosus (SLE), autoimmune thyroid disease, congestive heart
failure, pressure-overload cardiac hypertrophy, viral myocarditis,
myocardial ischemic injury, heart disease, artherogenesis,
atherosclerosis, cardiac events in diabetes, vascular disorders in
diabetes, muscle wasting, obesity, Alzheimer's disease, Parkinson's
disease, Huntington's disease, amyotrophic lateral sclerosis,
autoimmune thyroid disease, cachexia, Crohn's disease, inflammatory
bowel disease, sepsis, hepatitis B, transplantation rejection and
related immunology, retina degeneration, cataracts and
glaucoma.
[0348] In a further embodiment, the compounds of the application
are used for treating a disease or disorder associated with
inflammation in humans as well as other mammals. Exemplary
inflammatory conditions include, but not limited to rheumatoid,
arthritis, multiple sclerosis, degenerative joint disease,
spondouloarthropathies, osteoporosis, diabetes, Alzheimer's
disease, Parkinson's disease, shock, among others.
[0349] In a further embodiment, the compounds of the application
are used for treating a disease or disorder selected from allergies
and respiratory conditions, including asthma, bronchitis, pulmonary
fibrosis, allergic rhinitis, oxygen toxicity, emphysema, chronic
bronchitis, acute respiratory distress syndrome, and any chronic
obstructive pulmonary disease (COPD).
[0350] In a further embodiment, the compounds of the application
are used for treating a disease or disorder selected from viral
infections (HIV-1 and HIV-2), osteoporosis, osteoarthritis,
psoriasis, restenosis heart disease, diabetes-associated
cardiovascular disorders, inflammatory bowel disease, inflammatory
and autoimmune diseases (arthritis, psoriasis), seronegative
spondyloarthropathies (SpA), muscle wasting, obesity, allergy and
asthma, neurodegenerative disorders, including Alzheimer's (AD) and
Parkinson's (PD) diseases, and autoimmune diseases in a mammal
having or predisposed to said disease or disorder.
[0351] The introduction of a halogen atom into a molecule also
provides the opportunity for the use of the molecule in
radiolabeling applications. For example, .sup.18F is used as a
radiolabel tracer in the sensitive technique of Positron Emission
Tomography (PET). Accordingly the present application also includes
methods of using the compounds of Formula I for diagnostic and
imaging purposes, wherein the compounds of Formula I comprise at
least one radiolabel, such as .sup.18F.
[0352] Therefore the present application includes the use of one of
more compounds of the application for radiolabel imaging, wherein
the compounds of the application comprise at least one radiolabel,
such as .sup.18F.
[0353] The present application also includes a method of radiolabel
imaging comprising contacting a subject to be imaged with one or
more compounds of the application, and performing an imaging
technique on the subject, wherein the compounds of the application
comprise at least one radiolabel, such as .sup.18F. In an
embodiment the subject is a human or animal and the imaging
technique is PET and the one or more compounds of the application
or contacted with the subject by administration of a imaging
effective amount of the compound(s) to the subject.
[0354] The following non-limiting examples are illustrative of the
present application:
EXAMPLES
[0355] The introduction of the fluorine atom into molecules may
bring about changes in the physical and/or chemical properties of
the parent molecules, for example it may result in the enhancement
of pharmacokinetic properties and/or biological activities.
Replacement of hydrogen atoms may also result in improved thermal
and metabolic stability. Improved metabolic stability is generally
a desirable feature since the possibility exists that in vivo
decomposition may produce toxic effects. The properties of the
fluorine atom include its small size, low polarizability, high
electronegativity and its ability to form strong bonds with carbon.
Accordingly, bioactive compounds containing fluorinated groups such
as --OCHF.sub.2 are useful.
[0356] The geminal combination of an alkoxy or aryloxy group with a
fluorine atom offers the possibility of bonding/nonbonding
resonance, which can be formally expressed by the superposition of
a covalent and ionic limiting structure. This phenomenon, which
reveals itself as a lengthening and weakening of the carbon-halogen
bond and a shortening and strengthening of the carbon-oxygen bond
is known as the generalized anomeric effect [Schlosser et al. Chem.
Rev. 2005, 105, 827-856].
A. General Methods
[0357] All starting materials used herein were commercially
available or earlier described in the literature. The .sup.1H and
.sup.13C NMR spectra were recorded either on Bruker 300, Bruker
DPX400 or Varian +400 spectrometers operating at 300, 400 and 400
MHz for .sup.1H NMR respectively, using TMS or the residual solvent
signal as an internal reference, in deuterated chloroform as
solvent unless otherwise indicated. All reported chemical shifts
are in ppm on the delta-scale, and the fine splitting of the
signals as appearing in the recordings is generally indicated, for
example as s: singlet, br s: broad singlet, d: doublet, t: triplet,
q: quartet, m: multiplet. Unless otherwise indicated, in the tables
below, .sup.1H NMR data was obtained at 400 MHz, using CDCl.sub.3
as the solvent.
[0358] Purification of products was carried out using Chem Elut
Extraction Columns (Varian, cat #1219-8002), Mega BE-SI (Bond Elut
Silica) SPE Columns (Varian, cat #12256018; 12256026; 12256034) or
by flash chromatography in silica-filled glass columns.
B: Synthesis and Characterization of Compounds
I. Preparation of Intermediate Compounds of Formula II
[0359] Scheme V outlines the synthesis of intermediate compounds of
Formula IV(a) and Formula V(a) used in the preparation of the
intermediate compound of Formula II(a)(i), used in the preparation
of compounds of Formula I wherein R.sub.2 and/or R.sub.3 are a
--CH.sub.2--O--CHF.sub.2 moiety.
##STR00016##
[0360] Reagents and conditions used in Scheme V: (i)
2-fluorosulfonyldifluoroacetic acid, Cu(I)I, Na.sub.2SO.sub.4,
CH.sub.3CN 0.degree. C./30 min; (ii) H.sub.2, Pd/C (10%), THF, RT/2
hrs; (iii) 2M HCl/Et.sub.2O, 0.degree. C. to RT/2 hrs.
(a) Preparation of
(S)-2-tert-butoxycarbonylamino-3-difluoromethoxy-propionic acid
benzyl ester (XIII(a))
##STR00017##
[0362] To a stirred solution of the compound of Formula XII(a),
benzyl 2-(tert-butoxycarbonylamino)-3-hydroxypropanoate (1 g, 3.38
mmol) in acetonitrile (10 mL) was added copper(I) iodide (64.4 mg,
0.338 mmol) and sodium sulfate (48.1 mg, 0.338 mmol). The resulting
mixture was stirred at 60.degree. C. and treated with
2,2-difluoro-2-fluorosulfonylacetic acid (524 .mu.L, 5.08 mmol),
dropwise, as a solution in acetonitrile (2 mL) over a period of 1.5
h. Upon completion of the addition, the mixture was stirred for a
further 30 min then cooled to room temperature. The mixture was
diluted with diethyl ether and washed with brine (2.times.), water
(3.times.) and brine (lx). The organic phase was dried, filtered
and concentrated in vacuo then chromatographed in 0-30% ethyl
acetate in hexanes, to provide the compound of Formula XIII(a),
(S)-2-tert-butoxycarbonylamino-3-difluoromethoxy-propionic acid
benzyl ester (458 mg, 40%) as a colorless sticky oil. .sup.1H NMR
(300 MHz, CDCl.sub.3): .delta. (ppm) 7.32-7.38 (m, 5H), 6.18 (wt,
1H), 5.28 (dd, 1H), 5.19 (dd, 2H), 4.55 (dt, 1H) 4.22 (td, 1H),
4.15 (m, 2H), 1.38 (s, 9H).
[0363] Alternatively, the compound of Formula XIII(a),
(S)-2-tert-butoxycarbonylamino-3-difluoromethoxy-propionic acid
benzyl ester was prepared by difluoromethyl insertion of
(S)-2-tert-butoxycarbonylamino-3-thioformyloxy-propionic acid
benzyl ester in a 40-90% yield using
2,2-difluoro-1,3-dimethylimidazolidine, as a reagent for
difluoromethyl insertion.
(b) Preparation of
(S)-2-tert-butoxycarbonylamino-3-difluoromethoxy-propionic acid
(IV(a))
##STR00018##
[0365] A solution of the compound of Formula XIII(a),
(S)-2-tert-butoxycarbonylamino-3-difluoromethoxy-propionic acid
benzyl ester (1.76 g, 5.09 mmol) in THF was stirred with 10% Pd/C
(360 mg) under a hydrogen atmosphere for 2 hours. The reaction
mixture was filtered and concentrated to give the compound of
Formula IV(a),
(S)-2-tert-butoxycarbonylamino-3-difluoromethoxy-propionic acid
(1.3 g, 100%) as a sticky, colorless oil. .sup.1H NMR (300 MHz,
CDCl.sub.3): .delta. (ppm) 6.21 (wt, 1H), 5.35 (d, 1H), 4.59 (m,
1H), 4.35 (m, 1H), 4.21 (m, 1H) and 1.42 (s, 9H).
(c) Preparation of (S)-2-amino-3-difluoromethoxy-propionic acid
benzyl ester Hydrochloride salt (V(a))
##STR00019##
[0367] A solution of the compound of Formula XIII(a),
(S)-2-tert-butoxycarbonylamino-3-difluoromethoxy-propionic acid
benzyl ester (2.11 g, 5.89 mmol) in ether (10 mL) was treated with
2M HCl/ether, and stirred at 0.degree. C. for 2 hours. The reaction
mixture was concentrated to dryness and triturated with
hexane/ether to give the compound of Formula V(a)
(S)-2-amino-3-difluoromethoxy-propionic acid benzyl ester
hydrochloride salt (1.15 g, 69%) as a white powder. .sup.1H NMR
(300 MHz, CDCl.sub.3): b (ppm) 7.40-7.33 (m, 5H), 6.53 (broad s,
1H), 6.18 (wt, 1H), 5.25 (dd, 1H), 5.21 (dd, 2H), 4.53 (dt, 1H)
4.18 (td, 1H), 4.16-4.13 (m, 2H).
[0368] Scheme VI outlines the synthesis of intermediate compounds
of Formula III(a)(i) used in the preparation of the compounds of
Formula I wherein R.sub.2 and R.sub.3 are a
--CH.sub.2--O--CHF.sub.2 moiety.
##STR00020##
[0369] Reagents and conditions used in Scheme VI: (i) EDCl.HCl,
HOBt, N-methylmorpholine, CH.sub.2Cl.sub.2, 0.degree. C./ON; (ii)
H.sub.2, Pd/C (10%), THF, RT/1 hrs.
(d) Preparation of
(S)-2-((S)-2-tert-butoxycarbonylamino-3-difluoromethoxy-propionylamino)-3-
-difluoromethoxy-propionic acid benzyl ester (II(a)(ii))
##STR00021##
[0371] To a solution of the compound of Formula IV(a),
(S)-2-tert-butoxycarbonylamino-3-difluoromethoxypropionic acid
(1.15 g, 4.49 mmol) and the compound of Formula V(a),
(S)-2-amino-3-difluoromethoxypropionic acid benzyl ester
hydrochloride salt (1.15 g, 4.08 mmol), HOBt (0.689 g, 5.10 mmol)
and EDCl.HCl (0.867 g, 5.10 mmol) in dichloromethane (20 mL) was
added N-methylmorpholine (0.45 mL, 8.16 mmol) dropwise at 0.degree.
C. The reaction mixture was allowed to warm up to room temperature
and stirred overnight. The mixture was then diluted with ethyl
acetate and washed successively with water, 1N HCl and brine. The
organic layer was dried over MgSO.sub.4 and concentrated. The crude
product was purified by silica-gel column chromatography, eluting
with 10% to 12.5% ethyl acetate in hexanes, to give the compound of
Formula II(a)(ii),
(S)-2-((S)-2-tert-butoxycarbonylamino-3-difluoromethoxy-propionylamino)-3-
-di-fluoromethoxy-propionic acid benzyl ester (1.29 g, 65%) as an
off-white solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. (ppm)
7.35-7.41 (m, 5H), 7.18 (td, 1H), 6.18 (wt, 1H), 6.22 (wt, 1H),
5.25-5.20 (m, 1H), 5.19 (s, 2H), 4.82 (td, 1H), 4.42 (br s, 1H),
4.31 (td, 1H), 4.20-4.15 (m, 2H), 4.01 (dd, 1H), 1.41 (s, 9H).
[0372] In a similar manner, using the above general procedure, the
intermediate compounds shown in Table 1 were synthesized.
(e) Preparation of
(S)-2-((S)-2-tert-butoxycarbonylamino-3-difluoromethoxy-propionylamino)-3-
-difluoromethoxy-propionic acid (II(a)(i))
##STR00022##
[0374] A solution of the compound of Formula II(a)(ii),
(S)-2-((S)-2-tert-butoxycarbonylamino-3-difluoromethoxy-propionylamino)-3-
-difluoromethoxypropionic acid benzyl ester (1.27 g, 2.63 mmol) in
THF was stirred with 10% Pd/C (400 mg) under a hydrogen atmosphere
for 1 hour. The reaction mixture was filtered and concentrated to
give the compound of Formula II(a)(i),
(S)-2-((S)-2-tert-butoxycarbonylamino-3-difluoromethoxy-propionylamino)-3-
-difluoromethoxy-propionic acid (1.03 g, .about.100%) as a sticky,
off-white foam. NMR (d.sub.6-DMSO, 400 MHz) .delta. (ppm) 8.34 (d,
J=8 Hz, 1H), 7.12 (d, J=8 Hz, 1H), 6.66 (t, J=76 Hz, 1H), 6.62 (t,
J=76 Hz, 1H), 4.57-4.51 (m, 1H), 4.37-4.29 (m, 1H), 4.15-4.09 (m,
1H), 4.04-3.97 (m, 2H), 3.92-3.83 (m, 1H), 1.37 (s, 9H). Scheme VII
outlines the synthesis of compound intermediates of Formula IV(b)
and Formula V(b) which are used in the preparation of the compounds
of Formula (I) wherein R.sub.2 and/or R.sub.3 is a
--CH.sub.2--O--CH.sub.3 moiety.
##STR00023##
[0375] Reagents and conditions used in Scheme VII: (i)
BF.sub.4(OMe).sub.3, proton sponge, CH.sub.2Cl.sub.2, 0.degree. C.
to TR/ON; (ii) TFA, CH.sub.2Cl.sub.2, 0.degree. C./30 min; (iii)
HCl/Et.sub.2O.
(a) Preparation of
(S)-2-tert-butoxycarbonylamino-3-methoxy-propionic acid benzyl
ester (XIII(b))
##STR00024##
[0377] To a solution of the compound of Formula XII(a),
(S)-2-tert-butoxycarbonylamino-3-hydroxy-propionic acid benzyl
ester (1.7 g, 5.76 mmol) in dichloromethane (50 mL) was added
trimethyloxonium tetrafluoroborate (1.11 g, 7.50 mmol), followed by
the portion-wise addition of proton sponge (1.61 g, 7.50 mmol).
After stirring for 24 hrs at RT, the solution was concentrated, and
the residue was dissolved in ethyl acetate, and the solution
filtered through a pad of celite. The filtrate was washed with 1 M
HCl and brine. The organic layers were dried over MgSO.sub.4,
filtered, and concentrated in vacuo. Silica-gel flash
chromatography with 5% to 15% ethyl acetate in hexanes afforded the
compound of Formula XIII(b),
(S)-2-tert-butoxycarbonylamino-3-methoxy-propionic acid benzyl
ester (1.24 g, 69.6%) as a colorless oil. .sup.1H NMR (300 MHz,
CDCl.sub.3): .delta. (ppm) 7.35-7.29 (m, 5H), 4.90-4.87 (m, 1H),
4.51 (s, 2H), 3.92-3.87 (m, 1H), 3.80-3.78 (m, 1H), 3.56 (dd, 2H),
3.42 (dd, 1H), 3.33 (s, 3H), 1.42 (s, 9H).
(b) Preparation of
(S)-2-tert-butoxycarbonylamino-3-methoxy-propionic acid (IV(b))
##STR00025##
[0379] The compound of Formula IV(b) was prepared in a similar
manner as the compound of Formula IV(a), described above, from the
compound of Formula XIII(b)
(S)-2-tert-butoxycarbonylamino-3-methoxy-propionic acid benzyl
ester (0.76 g, 2.46 mmol), to provide the compound of Formula
IV(b), (S)-2-tert-butoxycarbonylamino-3-methoxy-propionic acid
(0.540 mg, 100%) as a sticky, colorless oil.
(c) Preparation of (S)-2-amino-3-methoxy-propionic acid benzyl
ester, hydrochloride salt (V(b))
##STR00026##
[0381] The compound of Formula V(b) was prepared, as a
hydrochloride salt, in a similar manner as the compound of Formula
V(a), described above, from the compound of Formula XIII(b),
(S)-2-tert-butoxycarbonylamino-3-methoxy-propionic acid benzyl
ester (0.80 g, 2.59 mmol), to provide the compound of Formula V(b),
(S)-2-amino-3-methoxy-propionic acid benzyl ester, hydrochloride
salt (100%) as a white powder.
[0382] Scheme VIII outlines the synthesis of the intermediate
compound of Formula II(b)(i) which is used in the preparation of
the compounds of Formula (I) wherein R.sub.2 is a
--CH.sub.2--O--CHF.sub.2 moiety and R.sub.3 is a
--CH.sub.2--O--CH.sub.3 group.
##STR00027##
[0383] Reagents and conditions used in Scheme VIII: (i) EDCl.HCl,
HOBt, DIPEA, CH.sub.2Cl.sub.2, 0.degree. C./ON; (ii) H.sub.2, Pd/C
(10%), THF, RT/2 hrs.
(d) Preparation of
(S)-2-((S)-2-tert-butoxycarbonylamino-3-difluoromethoxy-propionylamino)-3-
-methoxy-propionic acid benzyl ester (II(b)(ii))
##STR00028##
[0385] The compound of Formula II(b)(ii) was prepared as an
off-white foam in a similar manner as the compound of Formula
II(a)(ii), described above, from the compound of Formula IV(a),
(S)-2-tert-butoxycarbonylamino-3-difluoromethoxy-propionic acid and
the compound of Formula V(b), (S)-2-amino-3-methoxy-propionic acid
benzyl ester, hydrochloride salt.
(e) Preparation of
(S)-2-((S)-2-tert-butoxycarbonylamino-3-difluoromethoxy-propionylamino)-3-
-methoxy-propionic acid (II(b)(i))
##STR00029##
[0387] The compound of Formula II(b)(i) was prepared, as an
off-white sticky foam, in a similar manner as the compound of
Formula II(a)(i), described above from the compound of Formula
II(b)(ii),
(S)-2-((S)-2-tert-butoxycarbonylamino-3-difluoromethoxy-propionylamino)-3-
-methoxy-propionic acid benzyl ester.
[0388] Scheme IX outlines the synthesis of the intermediate of
Formula II(c)(i), used in the preparation of the compounds of
Formula (I) when R.sub.2 is a --CH.sub.2--O--CH.sub.3 group and
R.sub.3 is a --CH.sub.2--O--CHF.sub.2 moiety.
##STR00030##
[0389] Reagents and conditions used in Scheme IX: (i) EDCl.HCl,
HOBt, DIPEA, CH.sub.2Cl.sub.2, 0.degree. C./ON; (ii) H.sub.2, Pd/C
(10%), THF, RT/2 hrs.
[0390] In a similar manner to the above general procedure, the
other compounds shown in Table 2 were synthesized
II. Preparation of Intermediate Compounds of Formula III
[0391] Scheme X outlines the synthesis of intermediate epoxyketones
of Formula III used for the preparation of compounds of Formula I
wherein R.sup.4 is --CH.sub.2C.sub.6H.sub.5 and R.sup.5 is
methyl.
##STR00031##
[0392] Reagents and conditions used in Scheme X: (i) iBuOCOCl,
N-methylmorpholine, HNMe(OMe).HCl, TEA, CH.sub.2Cl.sub.2, 0.degree.
C./45 min; (ii) isopropenylmagnesium bromide, THF, 0.degree. C./2
hrs or 2-bromopropene, t-BuLi, Et.sub.2O, -78.degree. C./2 hrs;
(iii) (a) H.sub.2O.sub.2 (35%), benzonitrile, iPr.sub.2EtN, MeOH,
0.degree. C. to RT/ON; (b) Silica-gel column chromatography; (iv)
TFA, CH.sub.2Cl.sub.2, 0.degree. C./30 min.
[0393] In a similar manner as Scheme X, the synthesis of the
intermediate epoxyketones of Formula III(b) for compounds of
Formula I wherein R.sub.4 is CH.sub.2CH(CH.sub.3).sub.2 and R.sub.5
is methyl was prepared.
(a) Preparation of
[(S)-1-(methoxy-methyl-carbamoyl)-2-phenyl-ethyl]-carbamic acid
tert-butyl ester (VII(a))
##STR00032##
[0395] To a solution of
(S)-2-tert-butoxycarbonylamino-3-phenyl-propionic acid (24.85 g,
93.66 mmol) in dichloromethane (150 mL) was added
N-methylmorpholine (10.3 mL, 93.66 mmol), followed by addition of
isobutyl chloroformate (12.25 mL, 93.66 mmol) at 0.degree. C. The
reaction mixture was stirred for 20 min. then
N,O-dimethylhydroxylamine hydrochloride (9.14 g, 93.66 mmol) in one
portion was added. Subsequently, triethylamine (13 mL, 93.66 mmol)
was added dropwise over 15 min. The reaction mixture was stirred
for another hour, then it was quenched with 1N HCl (100 mL) and the
organic phase was washed with saturated NaHCO.sub.3 and brine (500
mL). The organic layers were dried over (MgSO.sub.4), filtered, and
concentrated in vacuo to give the Weinreb amide of Formula VII(a),
[(S)-1-(methoxy-methyl-carbamoyl)-2-phenyl-ethyl]-carbamic acid
tert-butyl ester as a clear sticky oil (29.3 g, 100%). .sup.1H NMR
(400 MHz, d.sub.6-DMSO): .delta. (ppm) 7.33-7.09 (m, 6H), 4.61-4.47
(m, 1H), 3.70 (s, 3H), 3.08 (s, 3H), 2.89-2.78 (m, 1H), 2.75-2.63
(m, 1H), 1.29 (s, 9H).
(b) Preparation of
((S)-1-benzyl-3-methyl-2-oxo-but-3-enyl)-carbamic acid tert-butyl
ester (VIII(a))
##STR00033##
[0397] To a 0.degree. C. solution of the above Weinreb amide of
Formula VII(a), (28.88 g, 93.66 mmol) in THF (150 mL) was added a
0.5 M solution in THF of isopropenyl magnesium bromide (386 mL,
192.9 mmol) at 0.degree. C. over 40 min. The reaction mixture was
then stirred at room temperature for 2 hours. The reaction mixture
was then quenched at 0.degree. C. with 1N HCl (350 mL). The aqueous
layer was extracted with EtOAc (2.times.200 mL). The organic layer
was washed successively with water and brine, dried over
MgSO.sub.4, filtered, and concentrated in vacuo to give, after
silica-gel flash chromatography with 5% to 10%
ethylacetate/hexanes, the compound of Formula VIII(a),
(S)-1-benzyl-3-methyl-2-oxo-but-3-enyl)-carbamic acid tert-butyl
ester (14.5 g, 53.5%) as a white powder. .sup.1H NMR (400 MHz,
d.sub.6-DMSO): .delta. (ppm) 7.33-7.09 (m, 6H), 4.61-4.47 (m, 1H),
3.70 (s, 3H), 3.08 (s, 3H), 2.89-2.78 (m, 1H), 2.75-2.63 (m, 1H),
1.29 (s, 9H).
(c) Preparation of
[(S)-1-benzyl-2-((S)-2-methyl-oxiranyl)-2-oxo-ethyl]-carbamic acid
tert-butyl ester and
[(S)-1-benzyl-2-((R)-2-methyl-oxiranyl)-2-oxo-ethyl]-carbamic acid
tert-butyl ester (IX(a))
##STR00034##
[0399] To a solution of the above compound of Formula VIII(a),
((S)-1-benzyl-3-methyl-2-oxo-but-3-enyl)-carbamic acid tert-butyl
ester (5.78 g, 20 mmol) in MeOH (250 mL) at 0.degree. C. was added
benzonitrile (15.46 mL, 150 mmol), H.sub.2O.sub.2 35% solution in
water (34.4 mL, 400 mmol), and diisopropylethylamine (26 mL, 150
mmol). The reaction mixture was stirred at 0.degree. C. to room
temperature overnight. The resulting mixture was concentrated under
reduced pressure to dryness. The obtained residue was quenched with
ice-water (100 mL) to provide a white precipitate. After
filtration, the aqueous layer was extracted with 20% ethyl acetate
in hexanes (2.times.200 mL). The organic layer was washed
successively with water and brine, dried over MgSO.sub.4, filtered,
and concentrated in vacuo to give, after silica-gel flash
chromatography with 3% to 3.5% ethyl acetate/hexanes the compound
of Formula S,R-IX(a),
[(S)-1-benzyl-2-((R)-2-methyl-oxiranyl)-2-oxo-ethyl]-carbamic acid
tert-butyl ester (3.33 g, 54%) as a white solid (.sup.1H NMR (300
MHz, CDCl.sub.3): .delta. (ppm) 7.21-7.29 (m, 3H), 7.17-7.21 (m,
2H), 4.92 (dd, 1H), 4.58 (td, 1H), 3.25 (d, 1H), 3.18 (dd, 1H),
2.94 (d, 1H), 2.75 (dd, 1H), 1.45 (s, 3H), 1.39 (s, 9H)), and the
compound of Formula S,S-IX(a),
[(S)-1-benzyl-2-((S)-2-methyl-oxiranyl)-2-oxo-ethyl]-carbamic acid
tert-butyl ester (1.66 g, 27%) as a sticky oil (.sup.1H NMR (300
MHz, CDCl.sub.3): .delta. (ppm) 7.30-7.22 (m, 3H), 7.20-7.15 (m,
2H), 4.95 (dd, 1H), 4.62 (td, 2H), 3.25 (d, 1H), 3.00 (dd, 1H),
2.82 (dd, 1H), 2.61 (dd, 2H), 1.45 (s, 3H), 1.40 (s, 9H)).
Preparation of
(S)-2-amino-1-((R)-2-methyl-oxiranyl)-3-phenyl-propan-1-one, TFA
salt (S,R-III(a))
##STR00035##
[0401] To a solution of (2.20 g, 7.20 mmol) of the compound of
Formula S,R-IX(a),
[(S)-1-benzyl-2-((R)-2-methyl-oxiranyl)-2-oxo-ethyl]-carbamic acid
tert-butyl ester in dichloromethane (10 mL), TFA (3.3 mL) was added
at 0.degree. C. The reaction mixture was stirred for 30 min. Excess
of TFA was evaporated to dryness, and the residue obtained was
triturated with 20% ether in hexanes (20 mL), followed by 100%
hexanes. After evaporation of solvents, drying under high vacuum
provided the compound of Formula S,R-III(a)
(S)-2-amino-1-((R)-2-methyl-oxiranyl)-3-phenyl-propan-1-one, TFA
salt (2.3 g, 100%), as an off-white powder. .sup.1H NMR (300 MHz,
methanol-d): .delta.(ppm) 7.22-7.44 (m, 5H), 4.32 (dd, 1H), 4.39
(dd, 1H), 3.17 (dd, 1H), 2.95 (dd, 1H), 2.88 (dd, 1H), 1.57 (s,
3H).
[0402] Alternatively, epoxyketones of the Formulae S,S-III(a) and
S,R-III(a) can be prepared as outlined in Schemes XI and XII.
##STR00036##
[0403] Reagents and conditions used in Scheme XI: (i) NaBH.sub.4,
CeCl.sub.3.7H.sub.2O, MeOH, THF, 0.degree. C./30 min; (ii) (a)
VO(acac).sub.2, t-BuO.sub.2H, CH.sub.2Cl.sub.2, 0.degree. C. to
RT/1 hr; (b) silica-gel column chromatography; (iii) (a)
Dess-Martin periodinane, CH.sub.2Cl.sub.2, 0.degree. C. to RT/2
hrs; (b) TFA, CH.sub.2Cl.sub.2, 0.degree. C./30 min.
##STR00037##
[0404] Reagents and conditions used in Scheme XII: (i) iBuOCOCl,
N-methylmorpholine, HNMe(OMe).HCl, TEA, CH.sub.2Cl.sub.2, 0.degree.
C./1 hr.; (ii) isopropenylmagnesium bromide, THF, 0.degree. C./2
hrs or 2-bromopropene, t-BuLi, Et.sub.2O, -78.degree. C./2 hrs.;
(iii) (a) H.sub.2O.sub.2 (35%), benzonitrile, i-Pr.sub.2EtN, MeOH,
0.degree. C. to RT/ON; (b) silica-gel column chromatography; (iv)
H.sub.2, Pd/C (10%), TFA, RT/6 hrs.
II. Preparation of Compounds of Formula I
Example 1
[0405] The preparation of the compound of Formula I of Example 1,
2-methyl-thiazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyloxiranyl)-2-oxo-ethylcarbamoyl-
]-2-difluoromethoxyethylcarbamoyl}-2-difluoromethoxyethyl)-amide is
outlined in Scheme XIII.
##STR00038##
[0406] Reagents and conditions used in Scheme XIII: (i) HBTU, HOBt,
DIPEA, THF, 0.degree. C. to RT/ON; (ii) H.sub.2, Pd/C (10%), THF,
RT/2 hrs.; (iii) HBTU, HOBt, DIPEA, THF,
2-methyl-thiazole-5-carboxylic acid, 0.degree. C. to RT/ON.
(a) Preparation of
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyl-oxiranyl)-2-oxo-ethylcarbamoy-
l]-2-difluoromethoxy-ethylcarbamoyl}-2-difluoromethoxy-ethyl)-carbamic
acid tert-butyl ester (XIV(a))
##STR00039##
[0408] To a solution of the compound of Formula II(b)(i),
(S)-2-((S)-2-tert-butoxycarbonylamino-3-difluoromethoxy-propionylamino)-3-
-methoxy-propionic acid (2.75 g, 7.028 mmol) and the compound of
Formula S,R-III(a),
(S)-2-amino-1-((R)-2-methyl-oxiranyl)-3-phenyl-propan-1-one, TFA
salt (12.04 g, 6.389 mmol), HOBt (1.04 g, 7.67 mmol), HBTU (2.91 g
7.67, mmol) in THF (75 mL) was added DIPEA (2.22 mL, 12.75 mmol)
dropwise at 0.degree. C. The reaction mixture was allowed to warm
up to room temperature and was stirred overnight. The mixture was
then quenched with ice water, washed with NaHCO.sub.3 and brine and
extracted with 2.times.100 mL of ethyl acetate. The organic layer
was dried over MgSO.sub.4 and concentrated. The crude product was
purified by column chromatography, eluting with 20% to 30% ethyl
acetate in hexanes, to give the title compound (2.7 g, 73%) as an
off-white solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. (ppm)
7.21-7.32 (m, 3H), 7.14 (dd, 2H), 6.85 (wd, 1H), 6.72 (wd, 1H),
6.18 (wt, 1H), 6.14 (wt, 1H), 5.26 (br s, 1H), 4.86 (td, 1H), 4.58
(td, 1H), 4.36 (m, 1H), 4.32-4.08 (m, 2H), 4.01 (dd, 1H), 3.90 (dd,
1H), 3.24 (d, 1H), 3.12 (dd, 1H), 2.90 (d, 1H), 2.85 (dd, 1H), 1.48
(s, 3H), 1.44 (s, 9H).
[0409] In a similar manner to the above general procedure, the
compounds shown in Table 3 were synthesized.
(b) Preparation of
(S)-2-amino-N--{(S)-1-[(S)-1-benzyl-2-((R)-2-methyl-oxiranyl)-2-oxo-ethyl-
carbamoyl]-2-difluoromethoxy-ethyl}-3-difluoromethoxy-propionamide
TFA salt (XV(a))
##STR00040##
[0411] To a solution of the compound of Formula XIV(a),
((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyl-oxiranyl)-2-oxo-ethylcarbamoy-
l]-2-difluoromethoxy-ethylcarbamoyl}-2-difluoromethoxy-ethyl)-carbamic
acid tert-butyl ester (2.68 g, 4.62 mmol) in dichloromethane (10
mL) at 0.degree. C. was added TFA (10 mL), and the resulting
mixture was stirred for 1 hr. Excess TFA was evaporated to dryness,
and the residue obtained was triturated with ether in hexanes
(2.times.20 mL) to provide the compound of Formula XV(a),
(S)-2-amino-N--{(S)-1-[(S)-1-benzyl-2-((R)-2-methyl-oxiranyl)-2-oxo-ethyl-
carbamoyl]-2-difluoromethoxy-ethyl}-3-difluoromethoxy-propionamide
TFA salt (2.34 g, 85%), as a pale-yellow solid. .sup.1H NMR (300
MHz, d.sub.6-DMSO): .delta. (ppm) 7.42-7.32 (m, 3H), 7.23 (dd, 2H),
6.85 (wd, 1H), 6.71 (br s, 1H), 6.68 (wd, 1H), 6.10 (w t, 1H), 5.98
(wt, 1H), 5.12 (br s, 1H), 4.78 (td, 1H), 4.51 (td, 1H), 4.29 (m,
1H), 4.30-3.99 (m, 2H), 3.97 (dd, 1H), 3.85 (dd, 1H), 3.21 (d, 1H),
3.10 (dd, 1H), 2.86 (d, 1H), 2.81 (dd, 1H), 1.45 (s, 3H).
[0412] In a similar manner to the above general procedure, the
compounds shown in Table 4 were synthesized.
(c) Preparation of 2-methyl-thiazole-5-carboxylic
acid((S)-1-{(S)-1-[(S)-1-benzyl-2-((R)-2-methyl-oxiranyl)-2-oxo-ethylcarb-
amoyl]-2-difluoromethoxy-ethylcarbamoyl}-2-difluoromethoxy-ethyl)-amide
(Example 1)
##STR00041##
[0414] To a solution of the compound of Formula XV(a),
(S)-2-amino-N--{(S)-1-[(S)-1-benzyl-2-((R)-2-methyl-oxiranyl)-2-oxo-ethyl-
carbamoyl]-2-difluoromethoxy-ethyl}-3-difluoromethoxy-propionamide
TFA salt (2.2 g, 3.70 mmol), 2-methyl-thiazole-5-carboxylic acid
(0.635 g, 4.44 mmol), HOBt (0.649 g, 4.81 mmol) and HBTU (1.82 g,
4.81 mmol) in THF (70 mL) was added DIPEA (1.29 mL, 7.4 mmol)
dropwise at 0.degree. C. The reaction mixture was allowed to warm
up to room temperature and was stirred overnight. The mixture was
then quenched with ice water, washed with NaHCO.sub.3 and brine and
extracted with 2.times.100 mL of ethyl acetate. The organic layer
was dried over magnesium sulphate and concentrated. The crude
product was purified by column chromatography, eluting with 70% to
80% ethyl acetate in hexanes, to give the compound of Formula I of
Example 1. (2.11 g, 94.6%) as a pale-yellowish solid. .sup.1H NMR
(300 MHz, CDCl.sub.3): .delta. (ppm) 8.10 (s, 1H), 7.28-7.25 (m,
3H), 7.20 (dd, 2H), 6.82 (d, 1H), 6.79 (d, 1H), 6.65 (d, 1H), 6.18
(wt, 1H), 6.14 (wt, 1H), 4.90-4.65 (m, 2H), 4.60 (td, 1H), 4.35
(dd, 1H), 4.25 (dd, 1H), 4.05 (td, 1H), 3.95 (dd, 1H), 3.30 (d,
1H), 3.28 (dd, 1H), 3.16 (d, 1H), 2.90 (dd, 1H), 2.85 (dd, 1H),
2.75 (s, 3H), 1.41 (s, 3H).
[0415] In a similar manner to the above general procedure, the
compounds of Formula I of Examples 2-24 shown in Table 5 were
synthesized.
Example 25
[0416] The compound of Formula I of Example 25,
(2S)-2-[[(2S)-3-(difluoromethoxy)-2-[(2-morpholinoacetyl)amino]propanoyl]-
amino]-N-[(1S)-3-methyl-1-[(2R)-2-methyloxirane-2-carbonyl]butyl]-3-phenyl-
-propanamide was prepared according to the following synthetic
procedure:
(a) Preparation of tert-butyl 2-morpholinoacetate
##STR00042##
[0418] To a stirred solution of tert-butyl bromoacetate (8.47 mL,
57.4 mmol) in THF (50 mL) was added a 1:1 mixture of triethylamine
(8 mL, 57.4 mmol) and morpholine (5.02 mL, 57.4 mmol), dropwise (a
mild exotherm was observed) and the resulting white suspension was
stirred at 60.degree. C. for 2 h. The mixture was diluted with
water (100 mL) and saturated sodium carbonate (50 mL) and extracted
with ethyl acetate (2.times.50 mL). The combined organics were
washed with saturated sodium carbonate (100 mL), water (3.times.50
mL), and brine (50 mL). The organic phase was dried over anhydrous
sodium sulfate, filtered and concentrated then chromatographed in
0%-100% ethyl acetate in hexanes. Product-containing fractions were
concentrated in vacuo giving the title product (11.5 g,
quantitative) as a pale yellow liquid. .sup.1H NMR (CDCl.sub.3, 400
MHz): .delta. (ppm) 3.78-3.71 (m, 4H), 3.10 (s, 2H), 2.59-2.54 (m,
4H), 1.46 (m, 9H).
(b) Preparation of 2-morpholinoacetic acid hydrochloride salt
##STR00043##
[0420] Tert-butyl 2-morpholinoacetate (11 g, 54.7 mmol) was stirred
with HCl, 4M in dioxane (54 mL), giving a white precipitate (a mild
exotherm was observed) which slowly dissolved with stirring at room
temperature. Ten minutes after complete dissolution, the mixture
solidified. Then the mixture was warmed to 60.degree. C. and the
thick suspension was stirred vigorously overnight. The mixture was
then cooled to room temperature, diluted with diethyl ether (60 mL)
and filtered to collect the title compound (8 g, 80%) as a white
solid. .sup.1H NMR (CD.sub.3OD, 400 MHz): .delta. (ppm) 4.13 (s,
2H), 3.94 (brm, 4H), 3.41 (brm, 4H).
(c) Preparation of benzyl
(2S)-2-(tert-butoxycarbonylamino)-3-phenyl-propanoate
##STR00044##
[0422] To a stirred solution of Boc-Phe-OH (10 g, 37.7 mmoL) in DMF
(50 mL) was added potassium carbonate (7.81 g, 56.5 mmol) followed
by benzyl chloride (4.55 mL, 39.6 mmol) and the resulting
suspension was stirred at 60.degree. C. for 3 d. The mixture was
diluted with water (400 mL) and extracted with ethyl acetate (75
mL, 2.times.25 mL). The combined organics were washed with brine
(200 mL), water (3.times.100 mL) and brine (50 mL). The organic
phase was dried over anhydrous sodium sulfate, filtered and
concentrated then chromatographed on silica gel eluting with 0%-20%
ethyl acetate in hexanes. The product-containing fractions were
concentrated in vacuo giving the title compound (12.84 g, 95%) as a
white solid.
[0423] In a similar manner to the above general procedure, benzyl
(2S)-2-(tert-butoxycarbonylamino)-4-methyl-pentanoate was
synthesized.
(d) Preparation of benzyl (2S)-2-amino-3-phenyl-propanoate
hydrochloride
##STR00045##
[0425] Benzyl (2S)-2-(tert-butoxycarbonylamino)-3-phenyl-propanoate
(12.8 g, 36 mmol) was stirred in HCl, 2 M in diethyl ether (90 mL)
and HCl, 4 M in dioxane (18 mL) and the resulting mixture was
stirred overnight at room temperature. The mixture was then diluted
with hexanes (100 mL) and filtered to collect the title compound
(7.98 g, 76%) as a fine white powder. .sup.1H NMR (CD.sub.3OD, 400
MHz): .delta. (ppm) 7.40-7.15 (m, 10H), 5.23 (s, 2H), 4.36-4.31 (m,
1H), 3.26-3.12 (m, 2H).
(e) Preparation of benzyl
(2S)-2-[[(2S)-2-(tert-butoxycarbonylamino)-3-(difluoromethoxy)propanoyl]a-
mino]-3-phenyl-propanoate (I(g)(ii))
##STR00046##
[0427]
(2S)-2-(tert-butoxycarbonylamino)-3-(difluoromethoxy)propanoic acid
(1.0 g, 3.92 mmol), benzyl (2S)-2-lamino-3-phenyl-propanoate (1.2
g, 4.11 mmol) and HOBt hydrate (0.66 g, 4.31 mmol) were stirred in
DCM (20 mL). The resulting mixture was treated with EDC
hydrochloride (0.83 g, 4.31 mmol), cooled to 0.degree. C. and
treated with DIPEA (0.715 mL, 4.11 mmol). The mixture was warmed to
room temperature and stirred overnight. The mixture was diluted
with DCM (30 mL) and washed with saturated sodium bicarbonate
(2.times.50 mL). The organic phase was dried over anhydrous sodium
sulfate, filtered and concentrated in vacuo then chromatographed in
0%-40% ethyl acetate in hexanes. The product-containing fractions
were concentrated in vacuo giving the compound of Formula II(g)(ii)
(1.9 g, 98%) as a waxy solid. .sup.1H NMR (CDCl.sub.3, 400 MHz):
.delta. (ppm) 7.40-6.95 (m, 10H), 6.71 (m, 1H), 6.14 (t, J=74 Hz,
1H), 5.19-5.09 (m, 2H), 4.95-4.85 (m, 1H), 4.40-4.20 (m, 2H),
3.98-3.90 (m, 1H), 3.19-3.10 (m, 2H), 1.44 (s, 9H).
(f) Preparation of benzyl
(2S)-2-[[(2S)-2-amino-3-(difluoromethoxy)propanoyl]amino]-3-phenyl-propan-
oate hydrochloride (XVI(a))
##STR00047##
[0429] Benzyl(2S)-2-[[(2S)-2-(tert-butoxycarbonylamino)-3
(difluoromethoxy)propanoyl]amino]-3-phenyl-propanoate (1.9 g, 3.86
mmol) was stirred in HCl, 2M in diethyl ether (12 mL), at room
temperature overnight. The resulting suspension was diluted with
hexanes and the filtered to collect the compound of Formula XVI(a)
(1.65 g, quantitative yield) as a white solid. The product was used
directly in the subsequent reaction.
(g) Preparation of benzyl
(2S)-2-[[(2S)-3-(difluoromethoxy)-2-[(2-morpholinoacetyl)
amino]propanoyl]amino]-3-phenyl-propanoate (XVII(a))
##STR00048##
[0431] To a stirred solution of benzyl
(2S)-2-[[(2S)-2-amino-3-(difluoromethoxy)propanoyl]amino]-3-phenyl-propan-
oate hydrochloride (1.65 g, 3.84 mmol), 2-morpholinoacetic acid
hydrochloride (768 mg, 4.23 mmol) and HOBt hydrate (648 mg, 4.23
mmol) in DCM (20 mL) was added EDC hydrochloride (811 mg, 4.23
mmol). The resulting mixture was cooled to 0.degree. C. and treated
with DIPEA (0.736 mL, 4.23 mmoL), dropwise, warmed to room
temperature and stirred overnight. The mixture was diluted with DCM
(30 mL) and washed with saturated sodium bicarbonate (2.times.50
mL). The organic phase was dried over anhydrous sodium sulfate,
filtered and concentrated in vacuo then chromatographed on silica
gel eluting with 0%-100% ethyl acetate in hexanes. The
product-containing fractions were concentrated in vacuo giving the
compound of Formula XVII(a) (1.54 g, 77%) as a white solid. .sup.1H
NMR (CDCl.sub.3, 400 MHz): .delta. (ppm) 7.72 (m, 1H), 7.35-6.90
(m, 10H), 6.12 (t, J=74 Hz, 1H), 5.14-5.04 (m, 2H), 4.81-4.75 (m,
1H), 4.65-4.55 (m, 1H), 4.26-4.20 (m, 1H), 3.93-3.87 (m, 1H),
3.65-3.50 (m, 4H), 3.15-2.90 (m, 4H), 2.45-2.35 (m, 4H).
[0432] In a similar manner to the above general procedure, the
compounds shown in Table 6 were synthesized.
(h) Preparation of
(2S)-2-[[(2S)-3-(difluoromethoxy)-2-[(2-morpholinoacetyl)
amino]propanoyl]amino]-3-phenyl-propanoic acid (XVIII(a))
##STR00049##
[0434] A stirred solution of benzyl
(2S)-2-[[(2S)-3-(difluoromethoxy)-2-[(2-morpholinoacetyl)amino]propanoyl]-
amino]-3-phenyl-propanoate (1.54 g, 2.96 mmol) in THF (10 mL) was
treated with palladium (10 wt. % on activated carbon) (157 mg,
0.148 mmol) and stirred under an atmosphere of hydrogen, balloon
pressure, for 1 h. The mixture was then filtered through a pad of
Celite.TM. and concentrated in vacuo giving the compound of Formula
XVIII(a) (1.27 g, quantitative yield) as colourless foam. .sup.1H
NMR (CDCl.sub.3, 400 MHz): .delta. (ppm) 8.09 (m, 1H), 7.24-7.05
(m, 5H), 6.10 (t, J=74 Hz, 1H), 5.6 (brs, 1H), 4.70-4.60 (m, 2H),
4.15-4.01 (m, 1H), 3.95-3.85 (m, 1H), 3.70-3.55 (m, 4H), 3.25-2.90
(m, 4H), 2.65-2.50 (m, 4H).
[0435] In a similar manner to the above general procedure, the
compounds shown in Table 7 were synthesized.
(i) Preparation of (2S)-2-[[(2S)-3-(difluoromethoxy)-2-[(2
morpholinoacetyl)amino]propanoyl]amino]-N-[(1S)-3-methyl-1-[(2R)-2-methyl-
oxirane-2-carbonyl]butyl]-3-phenyl-propanamide (Example-25)
##STR00050##
[0437] To a stirred solution of
(2S)-2-[[(2S)-3-(difluoromethoxy)-2-[(2-morpholinoacetyl)-amino]propanoyl-
] amino]-3-phenyl-propanoic acid (200 mg, 0.46 mmol),
(2S)-2-amino-4-methyl-1-(2-methyloxiran-2-yl)pentan-1-one
trifluoroacetate (133 mg, 0.46 mmol) and HOBt hydrate (71 mg, 0.46
mmol) in THF (10 mL) cooled to 0.degree. C. was added HBTU (176 mg,
0.46 mmol) followed by DIPEA (162 .mu.L, 0.93 mmol). The mixture
was warmed to room temperature and stirred for 4 h. The mixture was
diluted with DCM (40 mL) and washed with saturated sodium
bicarbonate (2.times.50 mL). The organic phase was dried over
anhydrous sodium sulfate, filtered and concentrated then
chromatographed on silica gel eluting with 50%-100% ethyl acetate
in hexanes. The product-containing fractions were concentrated in
vacuo and triturated with a 1:1 mixture of hexanes:diethyl ether
giving the compound of Formula I of Example 25 (108 mg, 40%) as a
pale solid. .sup.1H NMR (CDCl.sub.3, 400 MHz): .delta. (ppm) 7.78
(m, 1H), 7.31-7.15 (m, 5H), 6.71 (m, 1H), 6.22 (t, J=74 Hz, 1H),
6.10 (m, 1H), 4.65-4.49 (m, 3H), 4.30-4.20 (m, 1H), 4.09-4.00 (m,
1H), 3.75-3.63 (m, 4H), 3.25-2.80 (m, 5H), 2.55-2.45 (m, 4H), 2.17
(s, 3H), 1.55-1.18 (m, 3H), 0.95-0.85 (m, 6H).
[0438] In a similar manner to the above general procedure, the
compounds of Formula I of Examples 26-33 shown in Table 8 were
synthesized.
C. Biological Assays
Cells and Cell Culture
[0439] Human multiple myeloma cell lines 8226, H929, JJN3, KMH11,
KMS11, KMS18, LP1, MM1S, OPM2 and U266 were grown in Iscove
modified Dulbecco's medium (IMDM). Human leukemia cell lines K562,
OCI-AML2 and U937 were cultured in RPMI-1640 medium. Primary
peripheral blood mononuclear cells were isolated from multiple
myeloma patients by Ficoll density gradient centrifugation and bone
marrow aspirates were obtained from multiple myeloma patients at
the Princess Margaret Cancer Centre of the University Health
Network (UHN; Toronto, ON, Canada). Primary cells were cultured in
IMDM. The collection and use of human tissue for this study were
approved by the UHN institutional ethics review board. All cell
culture media were obtained from the Ontario Cancer Institute
Tissue Culture Media Facility (Toronto, ON, Canada) and were
supplemented with 10% fetal calf serum, 100 .mu.g/mL penicillin,
and 100 U/mL streptomycin (Hyclone, Logan, Utah). All cells were
grown in a humidified incubator at 37.degree. C. with 5%
CO.sub.2.
Proteasome Enzymatic Activity (Tumor Cell Lysates)
[0440] Cells were harvested by centrifugation at 1,200 rpm at room
temperature. Cell pellets were washed with PBS and lysed with assay
lysis buffer (50 mM HEPES
(N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid), pH 7.5; 150
mM NaCl; 1% Triton X-100; 2 mM ATP). Cell lysates were incubated on
ice for 30 minutes, mixed by vortex every 5 minutes, and then
centrifuged at 12,000 g for 10 minutes. The supernatant was
transferred to a 96-well plate. For each assay, 10 .mu.g of total
protein were incubated for 1 hour at 37.degree. C. with increasing
concentrations (1 nM to 10 .mu.M) of test compound diluted in assay
buffer (50 mM Tris-HCl (tris(hydroxymethyl)aminomethane-HCl), pH
7.5; 150 mM NaCl). DMSO alone was used as a control in every assay
plate. After incubation, a specific fluorogenic proteasome
substrate was added to each assay reaction at a final concentration
of 40 .mu.M in a total volume of 100 .mu.L.
N-Succinyl-Leu-Leu-Val-Tyr-7-amino-4-methylcoumarin
(Suc-Leu-Leu-Val-Tyr-AMC) was used for measuring chymotrypsin-like
(CT-L) activity,
t-butoxycarbonyl-Leu-Arg-Arg-7-amino-4-methylcoumarin
(Boc-Leu-Arg-Arg-AMC) for trypsin-like (T-L) activity, and
benzyloxycarbonyl-L-leucyl-L-leucyl-L-glutamyl-7-amino-4-methylcoumarin
(Z-Leu-Leu-Glu-AMC) for caspase-like (C-L) activity. The excitation
wavelength was set at 360 nm and the fluorescence emission
wavelength of AMC was detected at 460 nm. The fluorescence of free
AMC released during the enzymatic reaction was measured with a
SpectraMax M5 fluorescent spectrophotometric plate reader
(Molecular Devices, Sunnyvale, Calif.). AMC release rate was
measured at 37.degree. C. in a kinetic mode, recording every 5
minutes for 30 minutes. Experiments were performed in triplicate
and repeated at least twice.
Cell Viability Assays
[0441] Cellular viability was primarily assessed by
3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-
-2H-tetrazolium (MTS) assay according to the manufacturer's
instructions (Promega; Madison, Wis.). Cells were first seeded at a
density of 10,000 cells per well in tissue culture-treated 96-well
plates. Two hours after seeding, cells were treated with compounds
for 72 hours at concentrations as indicated. As a control, cells
were treated with DMSO alone in every assay plate. Following
treatment and the MTS assay, cell viability was independently
confirmed by reading the optical density (O.D.) at 490 nm and by
exclusion of trypan blue stain (Invitrogen; Burlington, ON,
Canada). The viability of primary mononuclear cells was determined
by staining with Annexin V.
[0442] Table 9 summarizes data obtained for representative
compounds of Formula I for cell viability and CT-L proteasome
activity using OCI-AML-2 and KMS-11 cell lines.
Proteases Purified from Thermoplasma acidophilum
[0443] Increasing concentrations (0, 0.26, 0.519, 1.0375, 2.075,
4.15 and 8.3 .mu.M) of compounds of Formula (I) and bortezomib were
incubated with 1 .mu.M purified beta subunits of proteasomes
isolated from Thermoplasma acidophilum in assay buffer supplemented
with 2 mM ATP. DMSO alone was used as a control. After 1 hour of
incubation at 37.degree. C., the Suc-Leu-Leu-Val-Tyr-AMC substrate
was added at a final concentration of 40 .mu.M and CT-L proteasome
activity was measured as described above for tumor cell
lysates.
[0444] FIG. 1 shows the effect of a representative of compounds of
Formula I (the compound of Example 1) on the enzymatic activity of
purified proteasomes, compared to bortezomib and shows that the
compound of Formula 1 of Example 1 disrupts the
ubiquitin-proteasome system. Purified beta proteasome subunits from
the archaebacteria Thermoplasma acidophilum (A) and whole cell
lysates from the human myeloma cell line LP1 (B) were used in the
study.
Immunoblotting Assay:
[0445] Whole cell lysates were prepared from LP1 cells in RIPA
lysis buffer. Anti-ubiquitin antibody was purchased from Cell
Signaling Technology Inc, (Danvers, Mass.) and anti-tubulin
antibody was purchased from Sigma-Aldrich (St. Louis, Mo.).
Secondary horseradish peroxidase-conjugated goat anti-mouse or
anti-rabbit IgG was purchased from Amersham Bioscience (Piscataway,
N.J.). Detection was performed using an enhanced chemiluminescence
kit from Pierce, (Rockford, Ill.).
[0446] LP1 cells exposed to compounds of Formula I demonstrated a
time-dependent and dose-dependent increase in the abundance of high
molecular weight ubiquitylated proteins detected by immunoblot,
using tubulin as a loading control bortezomib; IB, immunoblot.
Specifically, the compound of Examples 1 & 25 were tested and
showed an IC.sub.50<50 nM.
Combination Assays: Viability of Myeloma LP1 Cells in Culture
[0447] To examine whether compounds of Formula I can be combined
with other conventional and newer anti-multiple myeloma agents,
such as dexamethasone and bortezomib, respectively, LP1 myeloma
cells were treated for 72 hours with increasing concentrations of
compounds of Formula I in combination with dexamethasone or
bortezomib. After incubation, cell growth and viability was
measured by the MTS assay. The combination index (CI) analysis,
where a CI<1 indicates synergy between two drugs, a CI=1
indicates additivity, and a CI>1 indicates antagonism, was used
to determine whether the cytotoxicity produced by the combinations
with compounds of Formula I were synergistic, additive, or
antagonistic.
[0448] The effects of combining these agents with a representative
of compounds of Formula I i.e. the compound of Example 1 on the
viability of myeloma LP1 cells in culture were assessed. LP1 cells
were treated for 72 hours with increasing concentrations of the
compound of Example 1 in combination with dexamethasone or
bortezomib. After incubation, cell growth and viability was
measured by the MTS assay.
[0449] The combination of the compound of Example 1 and bortezomib
was synergistic with Combination Index values of 0.41, 0.46, and
0.88 at Fraction affected (Fa) levels of 0.39, 0.58, and 0.6,
respectively (FIG. 2A).
[0450] Thus using low doses of bortezomib may limit its off-target
activity and associated adverse effects while allowing for a more
efficient, synergistic, and specific blockade of CT-L proteasome
activity in combination with a compound of Formula 1 such as the
compound of Example 1.
[0451] The combination of the compound of Example 1 and
dexamethasone was synergistic with Combination Index values of
0.41, 0.44, and 0.5 at Fraction affected levels of 0.43, 0.5, and
0.64, respectively (FIG. 2B).
[0452] The data provides the rationale for combining the compound
of Example 1 with another agent such as bortezomib to achieve
useful proteasome inhibition and antitumor activity, which may
allow for use of lower doses of agents such as bortezomib and
potentially reduced side effects.
Cell Death in Primary Myeloma Cells Preferential Over Normal
Hematopoietic Cells:
[0453] The viability of leukemia and myeloma cells treated with
compounds of Formula I or bortezomib was assessed with the use of a
CellTiter 96.TM. AQueous One Solution Cell Proliferation Assay
(Promega, Madison, Wis.), which is a form of the
3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-
-2H-tetrazolium inner salt (MTS) assay, or with fluorescence-based
Alamar Blue cell viability reagent (Invitrogen, Carlsbad, Calif.)
according to manufacturer's instructions and as described
previously [J. Natl. Cancer Inst. 2007, 99(10), 811-822; Hum.
Reprod. 2007, 22(5), 1304-1309] or by trypan blue staining.
[0454] Apoptosis was measured by staining cells treated with
compounds of Formula I with annexin V-fluorescein isothiocyanate
and propidium iodide (both from Biovision Research Products,
Mountain View, Calif.) and flow cytometry according to
manufacturer's instructions and as previously described [Blood.
2005, 105(10), 4043-4050]. Experiments were performed at least in
duplicate and repeated at least twice (n=4-20 data points). Viable
primary myeloma cells were identified by staining with
phycoerythrin-conjugated mouse monoclonal anti-CD138 antibody (20
.mu.L/106 cells; Beckman Coulter, Brea, Calif.). The percentage of
myeloma cells that were CD138 positive and annexin V negative after
compounds of Formula (I) treatment compared with untreated samples
was quantified as a marker of cell viability as previously
described [Blood. 2007, 109(12), 5430-5438].
[0455] Effects of a representative of compounds of Formula I (the
compound of Example 1) on the viability of primary malignant and
normal hematopoietic cells isolated from patient samples was also
studied. The mononuclear cells from a bone and marrow of a patient
with myeloma were incubated for 24 hours with increasing
concentrations of the compound of Example 1 (Patient 1); the
peripheral blood mononuclear cells from a patient with plasma cell
leukemia were incubated for 24 hours with increasing concentrations
of the compound of Example 1 (Patient 2); and a primary myeloma
patient's samples were treated with increasing concentrations of
the compound of Example 1 (Patient 3). Viability of normal
hematopoietic cells (CD138-) and myeloma cells (CD138+) were
measured by flow cytometry with PE-labeled anti-CD138 and
FITC-labeled Annexin V co-staining.
[0456] Data represent the percentage of viable cells from each
patient sample (FIG. 3). FIG. 3 shows the effects of the compound
of Example 1 on the cell viability of primary samples from myeloma
patients detected by Annexin V staining and flow cytometry
following 24 hours of treatment, relative to vehicle control. The
compound of Example 1 selectively reduced the viability of primary
CD138+ myeloma cells over CD138- normal hematopoietic cells
isolated from the bone marrow of patients with multiple myeloma
(FIGS. 3A and 3C) or from the peripheral blood of a patient with
plasma cell leukemia (FIG. 3B). The compound of Example 1 induced
cell death in the plasma cells of the myeloma patients at nanomolar
concentrations. The compound of Example 1 was less cytotoxic to the
normal mononuclear hematopoietic cells with an LD.sub.50>2.5
.mu.M. By contrast, carfilzomib demonstrates a much narrower
therapeutic index between CD138+ and CD-138- in patient samples
[Trudel et al. ASH 2009, Poster Board 1-867].
Murine Red Blood Cells and Organ Homogenates
[0457] All mouse experiments were performed in accordance with
approval from the Ontario Cancer Institute institutional animal
review board. Five- to six-week-old male non-obese diabetic/severe
combined immunodeficient (NOD/SCID) mice were grouped randomly (n=3
mice per group). Mice were administered vehicle (5% DMSO, 20%
Cremophor) or compounds of Formula I at different doses either
intravenously or by oral gavage, and venous blood samples (20-50
.mu.L) were collected from each mouse over 24 hours. Blood samples
were mixed with heparin (APP Pharmaceuticals; Schaumburg, Ill.) in
0.5 mL tubes in accordance with the manufacturer's instructions.
After centrifugal separation at 3,000 g for 10 minutes, red blood
cells (RBCs) in the bottom layer were transferred into a new tube
and stored at -70.degree. C. until use. RBCs were lysed with assay
lysis buffer and incubated on ice for 30 minutes, mixing by vortex
every 5 minutes, and then centrifuged at 12,000 g for 10 minutes.
The supernatant was transferred to a 96-well plate and proteasome
activity was measured as described in above for tumor cell
lysates.
[0458] To evaluate proteasome activity of compounds of Formula I in
the organs of treated mice, five- to six-week-old male NOD/SCID
mice were sacrificed by CO.sub.2 inhalation 4 hours after oral
gavage with control vehicle and administered representative of
compounds of Formula I at doses of 30 mg to 100 mg. The brain,
liver, heart, lung, kidney, femurs and bone marrow were removed,
washed with PBS, and stored at -70.degree. C. until use. Prior to
analysis, mouse organs were thawed and homogenized on ice in assay
lysis buffer. Femurs were cut at both ends and bone marrow was
flushed out with assay lysis buffer. Organ homogenates were
centrifuged at 13,000 g for 30 minutes at 4.degree. C. and the
supernatant was used for measuring proteasome activity as described
above for tumor cell lysates.
[0459] Proteasome subunit activity (Chymotrypsin-like,
Trypsin-like, Caspase-like, CT-L, T-L, C-L, respectively) were
monitored over the course of 24 hrs following oral administration
of representative compounds of Formula I. NOD/SCID mice were
treated with the compound from Example 1 (50 mg/kg by oral gavage)
and the compound from Example 26 (60 mg/kg by oral gavage) or
vehicle control for up to 24 hours, as described above.
[0460] Table 10 summarizes the data for the representative
compounds of Formula (I); the compounds of Examples 1 and 26. As
can be seen from the data in Table 10, the compounds of Examples 1
and 26 display pharmacodynamic activity following oral
administration to mice.
National Cancer Institute (NCI) Screening Panel:
[0461] Representative compounds of Formula I were screened using
the National Cancer Institute (NCI) screening panel, which consists
of a panel of 60 different human tumor cell lines, representing
leukemia [CCRF-CEM, HL-60 (TB), K-562, MOLT-4, SR], melanoma [LOX
IMVI, MALME-3M, M14, SMDA-MB-435, SK-MEL-2, SK-MEL-28, SK-MEL-5,
UACC-257 and UACC-62] and cancers of the lung [A549/ATCC, EKVX,
HOP-62, HOP-93, NCI-H226, NCI-H23, NCI-H322M, NCI-H460], colon
[COLO 205, HCT-116, HCT-15, HT29, KM12, SW-620], brain [SF-268,
SF-295, SF-539, SNB-19, SNB-75, U251], ovary [IGROV1, OVCAR-3,
OVCAR-4, OVCAR-5, OVCAR-8, NCI/ADR-RES, SK-OV-3], breast [MCF7,
MDA-MB-231, BT-549, T-47D, MDA-MB-468], prostate [PC-3, DU-145],
and renal [786-0, A498, ACHN, CAKI-1, RXF-393, SN12C, TK-10, UO-31]
cancers.
[0462] After 24 h, two plates of each cell line are fixed in situ
with TCA, to represent a measurement of the cell population for
each cell line at the time of drug addition (T.sub.z). Experimental
drugs are solubilised in dimethyl sulfoxide at 400-fold the desired
final maximum test concentration and stored frozen prior to use. At
the time of drug addition, an aliquot of frozen concentrate is
thawed and diluted to twice the desired final maximum test
concentration with complete medium containing 50 .mu.g/ml
gentamicin. Additional four, 10-fold or 1 log serial dilutions are
made to provide a total of five drug concentrations plus control.
Aliquots of 100 .mu.l of these different drug dilutions are added
to the appropriate microtiter wells already containing 100 .mu.l of
medium, resulting in the required final drug concentrations.
[0463] Following drug addition, the plates are incubated for an
additional 48 h at 37.degree. C., 5% CO.sub.2, 95% air, and 100%
relative humidity. For adherent cells, the assay is terminated by
the addition of cold TCA (trichloroacetic acid). Cells are fixed in
situ by the gentle addition of 50 .mu.l of cold 50% (w/v) TCA
(final concentration, 10% TCA) and incubated for 60 minutes at
4.degree. C. The supernatant is discarded, and the plates are
washed five times with tap water and air dried. Sulforhodamine B
(SRB) solution (100 .mu.l) at 0.4% (w/v) in 1% acetic acid is added
to each well, and plates are incubated for 10 minutes at room
temperature. After staining, unbound dye is removed by washing five
times with 1% acetic acid and the plates are air dried. Bound stain
is subsequently solubilised with 10 mM trizma base, and the
absorbance is read on an automated plate reader at a wavelength of
515 nm. For suspension cells, the methodology is the same except
that the assay is terminated by fixing settled cells at the bottom
of the wells by gently adding 50 .mu.l of 80% TCA (final
concentration, 16% TCA). Using the seven absorbance measurements
[time zero, (T.sub.i), control growth, (C), and test growth in the
presence of drug at the five concentration levels (T.sub.i)], the
percentage growth is calculated at each of the drug concentration
levels. Percentage growth inhibition is calculated as:
[(T.sub.i-T.sub.z)/(C-T.sub.z)].times.100 for concentrations in
which T.sub.i>/=T.sub.z and
[(T.sub.i-T.sub.z)/T.sub.z].times.100 for concentrations in which
T.sub.i<T.sub.z.
[0464] Three dose response parameters are calculated for each
experimental agent. Growth inhibition of 50% (GI.sub.50) is
calculated from [(T.sub.i-T.sub.z)/(C-T.sub.z)].times.100=50, which
is the drug concentration resulting in a 50% reduction in the net
protein increase (as measured by SRB staining) in control cells
during the drug incubation. The drug concentration resulting in
total growth inhibition (TGI) is calculated from T.sub.i=T.sub.z.
The LC.sub.50 (concentration of drug resulting in a 50% reduction
in the measured protein at the end of the drug treatment as
compared to that at the beginning) indicating a net loss of cells
following treatment is calculated from
[(T.sub.i-T.sub.z)/T.sub.z].times.100=-50. Values are calculated
for each of these three parameters if the level of activity is
reached. However, if the effect is not reached or is exceeded, the
value for that parameter is expressed as greater or less than the
maximum or minimum concentration tested.
[0465] The results obtained from this study shows compounds of
Formula I are effective against the cell lines of the 60 human
tumor cell lines panel. Inhibition of human cancer cell lines in
vitro by representative compounds of Formula (I) are shown in Table
11 (Example 1), Table 12 (Example 25), Table 13 (Example 26) and
Table 14 (Example 29).
Inhibition of Tumor Growth in Mouse Xenograft Models:
Efficacy on Tumor Growth in Hollow Fiber Assay (HFA)
[0466] Hollow fibre assay in vivo pharmacodynamic studies were
carried out. This in vivo animal model uses semi-permeable
biocompatible fibres that are filled with cancer cells, heat-sealed
and implanted surgically (s.c. or i.p.) in mice or rats, which can
then be treated with chemotherapeutics. Many different cell lines
from different tissue origins and cellular characteristics can be
encapsulated within the fibres, providing a cost-effective
screening method.
[0467] The Oncology Program Screening and Characterization Strategy
group at NCI uses HFA, which provides quantitative indices of drug
efficacy with minimal expenditures of time and materials. Thus, the
HFA is being utilized as the initial in vivo experience for
compounds of Formula I.
[0468] A total of 3 different tumor lines are prepared for each
experiment so that each mouse receives 3 intraperitoneal implants
(1 of each tumor line) and 3 subcutaneous implants (1 of each tumor
line). A compound is considered for xenograft testing if it
produces cell kill of any cell line at either dose level evaluated,
or it has a combined ip+sc score of 20 or greater, a sc score of 8
or greater. This scoring system has been validated by DCTDC
statisticians in CTEP to represent a level of detection expected to
score current "standard" agents as active.
[0469] The compound of Example 1 was evaluated in an HFA assay via
ip administration at two doses; a 75 mg/kg/dose and a 35.5
mg/kg/dose, QD.times.4. A standard panel of 12 tumor cell lines was
used for hollow fiber screening of compound of Formula I. These
include NCI-H23 and NCI-H522 for non-small cell lung cancer,
MDA-MB-231 for breast cancer, MDA-MB-435, LOX IMVI and UACC-62 for
melanoma, SW-620 and COLO 205 for colon cancer, OVCAR-3 and OVCAR-5
for ovarian cancer, and U251 and SF-295 for CNS (central nervous
system) cancer. The compound of Example 1 of Formula I produced
cell killing of multiple cell lines at either of the two doses (75
mg/kg/dose and 35.5 mg/kg/dose).
Efficacy on Tumor Growth in Leukemia AML2 Xenograft Models:
[0470] Sublethally irradiated NOD-SCID mice bearing an established
human tumor xenograft derived from RL and BALB/c mice challenged
with the murine tumor cell line AML2 were treated twice weekly on
days 1 and 2 (QD.times.2) with a dose of 30 mg/kg to 200 mg/kg of
the compound of Example 1 by oral gavage. Dosing began on Day 6
post tumor challenge. Results demonstrated that compounds of
Formula I exhibit antitumor response. For example, the compound of
Example 1 of Formula I exhibits an antitumor response of greater
than 40% tumor growth inhibition in the AML2 mouse xenograft
leukemia model at 30 mg/kg with no observed gross adverse effects
including reductions in body weight or alterations in
behaviour.
Efficacy on Tumor Growth Inhibition in Myeloma MM.1S Mouse
Xenograft Models:
[0471] The mice were irradiated (200 rads) using a Co60 irradiator
source. After 24 hrs, each mouse was inoculated subcutaneously with
5.times.10.sup.6 MM.1S tumor cells in 0.1 ml PBS for tumor
development. Treatments were started when the tumor volume reached
100 mm.sup.3. Each treatment group consisted of 10 mice. The
compound of Example 1 was administrated to the tumor-bearing mice
according to a specific predetermined regimen. The compound from
Example 1 at dose levels of 50 mg/kg (p.o, qd.times.28 days) and
100 mg/kg (p.o, days 1, 3, 5/wk.times.4 wks) produced statically
significant antitumor activity vs. control with no observed gross
adverse effects including reductions in body weight or alterations
in behaviour.
[0472] While the present application has been described with
reference to what are presently considered to be the preferred
examples, it is to be understood that the present application is
not limited to the disclosed examples. To the contrary, the present
application is intended to cover various modifications and
equivalent arrangements included within the spirit and scope of the
appended claims.
[0473] All publications, patents and patent applications are herein
incorporated by reference in their entirety to the same extent as
if each individual publication, patent or patent application was
specifically and individually indicated to be incorporated by
reference in its entirety. Where a term in the present application
is found to be defined differently in a document incorporated
herein by reference, the definition provided herein is to serve as
the definition for the term.
TABLE-US-00001 TABLE 1 Compound Yield # Structure Nomenclature
Appearance (%) II(b)(ii) ##STR00051## Benzyl-(2S)-2-[[(2S)-2-(tert-
butoxycarbonylamino)-3- (difluoromethoxy)propanoyl]-
amino]-3-methoxy- propanoate off-white powder 71 .sup.1H NMR (300
MHz CDCl.sub.3): .delta. (ppm): 7.41-7.39 (m, 5H), 7.18 (td, 1H),
6.21 (wt, 1H), 5.25-5.20 (m, 1H), 5.19 (s, 2H), 4.82 (td, 1H), 4.42
(br s, 1H), 4.31 (td, 1H), 4.20-4.15 (m, 2H), 4.01 (dd, 1H), 3.25
(s, 3H), 1.41 (s, 9H). II(c)(ii) ##STR00052##
Benzyl-(2S)-2-[[(2S)-2-(tert- butoxycarbonylamino)-3-
methoxy-propanoyl]amino)- 3- (difluoromethoxy)propanoate off-white
powder 86 .sup.1H NMR (300 MHz CDCl.sub.3): .delta. (ppm):
7.42-7.38 (m, 5H), 7.20 (td, 1H), 6.23 (wt, 1H), 5.22-5.26 (m, 1H),
5.18 (s, 2H), 4.79 (td, 1H), 4.41 (br s, 1H), 4.33 (td, 1H),
4.19-4.16 (m, 2H), 4.03 (dd, 1H), 3.26 (s, 3H), 1.43 (s, 9H).
II(d)(ii) ##STR00053## Benzyl-(2S)-2-[[(2S)-2-(tert-
butoxycarbonylamino)-3- (difluoromethoxy)propanoyl]
amino]-4-methyl-pentanoate white solid 76 .sup.1H NMR (CDCl.sub.3,
400 MHz): .delta. 7.42-7.35 (m, 5H), 6.87-6.81 (m, 1H), 6.13 (t, J
= 74 Hz, 1H), 5.22-5.17 (m, 2H), 4.89-4.77 (m, 2H), 4.33- 4.25 (m,
1H), 4.18-4.09 (m, 2H), 1.75-1.61 (m, 2H), 1.54-1.39 (m, 1H),
0.99-0.87 (m, 6H). II(e)(ii) ##STR00054##
Benzyl-(2S)-2-[[(2S)-2-(tert- butoxycarbonylamino)-4-
methyl-pentanoyl]amino]-3- (difluoromethoxy)propanoate clear sticky
oil 85 .sup.1H NMR (CDCl.sub.3, 400 MHz): .delta. 7.40-7.31 (m,
5H), 6.89-6.83 (m, 1H), 6.14 (t, J = 74 Hz, 1H), 5.24-5.18 (m, 2H),
4.87-4.79 (m, 2H), 4.32- 4.26 (m, 1H), 4.18-4.09 (m, 2H), 1.73-1.62
(m, 2H), 1.53-1.40 (m, 1H), 0.98-0.88 (m, 6H). II(f)(ii)
##STR00055## benzyl (2S)-2-[[(2S)-2-(tert- butoxycarbonylamino)-3-
phenyl-propanoyl]amino]-3- (difluoromethoxy)propanoate white solid
94 II(g)(ii) ##STR00056## Benzyl-(2S)-2-[[(2S)-2-(tert-
butoxycarbonylamino)-3- (difluoromethoxy)propanoyl]
amino]-3-phenylpropanoate white solid 78
TABLE-US-00002 TABLE 2 Compound Yield # Structure Nomenclature
Appearance (%) II(b)(i) ##STR00057## (2S)-2-[[(2S)-2-(tert-
butoxycarbonylamino)-3- (difluoromethoxy)propanoyl]-
amino]-3-methoxypropanoic acid off-white foam 98 NMR (d.sub.6-DMSO,
400 MHz): .delta. 8.36 (d, J = 8 Hz, 1H), 7.12 (d, J = 8 Hz, 1H),
6.63 (t, J = 76 Hz, 1H), 4.57-4.51 (m, 1H), 4.37-4.29 (m, 1H),
4.15-4.09 (m, 1H), 4.04-3.97 (m, 2H), 3.92-3.83 (m, 1H), 3.21 (s,
3H), 1.37 (s, 9H). II(c)(i) ##STR00058## (2S)-2-[[(2S)-2-(tert-
butoxycarbonylamino)-3- methoxy-propanoyl]amino]-
3(difluoromethoxy)propanoic acid clear sticky oil 99 NMR
(d.sub.6-DMSO, 400 MHz): .delta. 8.33 (d, 1H), 7.14 (d, J = 8 Hz,
1H), 6.63 (t, J = 75 Hz, 1H), 4.59-4.49 (m, 1H), 4.38-4.27 (m, 1H),
4.17- 4.10 (m, 1H), 4.06-3.97 (m, 2H), 3.95-3.80 (m, 1H), 3.22 (s,
3H), 1.38 (s, 9H). II(f)(i) ##STR00059## (2S)-2-[[(2S)-2-(tert-
butoxycarbonylamino)-3- phenyl-propanoyl]amino]-3-
(difluoromethoxy)propanoic acid white solid 100 II(g)(i)
##STR00060## (2S)-2-[[(2S)-2-(tert- butoxycarbonylamino)-3-
(difluoromethoxy)propanoyl] amino]-3-phenyl-propanoic acid white
powder 98 II(d)(i) ##STR00061## (2S)-2-[[(2S)-2-(tert-
butoxycarbonylamino)-3- (difluoromethoxy)propanoyl]
amino]-4-methyl-pentanoic acid off-white yellowish foam 66 .sup.1H
NMR (d.sub.6-DMSO, 400 MHz): 8.35 (d, 1H), 6.11 (t, J = 74 Hz, 1H),
5.17-5.05 (m, 1H), 4.79-4.71 (m, 1H), 4.29-4.16 (m, 1H), 4.11-4.01
(m, 1H), 1.68-1.52 (m, 2H), 1.50-1.27 (m, 1H), 1.38 (s, 9H), 0.91-
0.83 (m, 6H). II(e)(i) ##STR00062## (2S)-2-[[(2S)-2-(tert-
butoxycarbonylamino)-4- methyl-pentanoyl]amino]-3-
(difluoromethoxy)propanoic acid clear oil 85 .sup.1H NMR
(d.sub.6-DMSO, 400 MHz): 8.37 (d, 1H), 6.13 (t, J = 74 Hz, 1H),
5.15-5.01 (m, 1H), 4.81-4.75 (m, 1H), 4.28-4.15 (m, 1H), 4.13-4.02
(m, 1H), 1.71-1.55 (m, 2H), 1.52-1.26 (m, 1H), 1.38 (s, 9H), 0.94-
0.82 (m, 6H).
TABLE-US-00003 TABLE 3 Compound Formula and Yield # Structure
Nomenclature Appearance (%) XIV(b) ##STR00063##
C.sub.25H.sub.35F.sub.2N.sub.3O.sub.8 ((S)-1-{(S)-1-[(S)-1-
Benzyl-2-((R)-2-methyl- oxiranyl)-2-oxo- ethylcarbamoyl]-2-
methoxyethylcarbamoyl}-2- difluoromethoxyethyl)- carbamic acid
tert-butyl ester off-white solid 85 .sup.1H-NMR (300 MHz,
CDCl.sub.3): .delta. 7.21-7.35 (m, 3H), 7.18 (dd, 2H), 6.80 (wd,
1H), 6.72 (wd, 1H), 6.21 (w t, 1H), 5.29 (br s, 1H), 4.79 (td, 1H),
4.58 (td, 1H), 4.36 (m, 1H), 4.35-4.12 (m, 2H), 4.10 (dd, 1H), 3.87
(dd, 1H), 3.26 (d, 1H), 3.25 (s, 3H), 3.14 (dd, 1H), 2.87 (d, 1H),
2.87 (dd, 1H), 1.49 (s, 3H), 1.43 (s, 9H). XIV(c) ##STR00064##
C.sub.25H.sub.35F.sub.2N.sub.3O.sub.8 ((S)-1-{(S)-1-[(S)-1-
Benzyl-2-((R)-2-methyl- oxiranyl)-2-oxo- ethylcarbamoyl]-2-
difluoromethoxyethyl- carbamoyl}-2- methoxyethyl)-carbamic acid
tert-butyl ester off-white powder 83 .sup.1H-NMR (300 MHz,
CDCl.sub.3): .delta. 7.20-7.28 (m, 3H), 7.20 (dd, 2H), 6.83 (wd,
1H), 6.75 (wd, 1H), 6.71 (w t, 1H), 5.27 (br s, 1H), 4.81 (td, 1H),
4.60 (td, 1H), 4.38 (m, 1H), 4.38-4.09 (m, 2H), 4.12 (dd, 1H), 3.91
(dd, 1H), 3.29 (d, 1H), 3.26 (s, 3H), 3.16 (dd, 1H), 2.84 (d, 1H),
2.81 (dd, 1H), 1.50 (s, 3H), 1 41 (s, 9H). XIV(d) ##STR00065##
C.sub.22H.sub.35F.sub.4N.sub.3O.sub.8 (S)-2-Difluoromethoxy-
1-{(S)-2-di- fluoromethoxy-1-[(S)-3- methyl-1-((R)-2-methyl-
oxiranecarbonyl)- butylcarbamoyl]- ethylcarbamoyl}-ethyl)- carbamic
acid tert-butyl ester off-white powder 99 .sup.1H-NMR (CDCl.sub.3,
300 MHz): .delta. 7.02 (d, J = 6 Hz, 1H), 6.74-6.64 (m, 1H), 6.25
(t, J = 75 Hz, 1H), 6.21 (t, J = 75 Hz, 1H), 5.34-5.24 (m, 1H),
4.69-4.58 (m, 3H), 4.44-4.24 (m, 4H), 4.10-4.04 (m, 1H), 3.99- 3.92
(m, 1H), 3.24 (d, J = 6 Hz, 1H), 2.91-2.87 (m, 2H), 1.82-1.72 (m,
1H), 1.51 (s, 3H), 1.46 (s, 9H), 0.96-0.90 (m, 6H). XIV(e)
##STR00066## C.sub.24H.sub.41F.sub.2N.sub.3O.sub.7
((S)-2-Difluoromethoxy- 1-{(S)-3- methyl-1-[(S)-3-methyl-
1-((R)-2-methyl- oxiranecarbonyl)- butylcarbamoyl]-
butylcarbamoyl}-ethyl)- carbamic acid tert-butyl ester white powder
70 NMR (CDCl.sub.3, 300 MHz): .delta. 6.98 (d, 1H), 6.74-6.64 (m,
1H), 6.22 (t, J = 75 Hz, 1H), 5.33-5.24 (m, 1H), 4.69-4.58 (m, 3H),
4.44-4.24 (m, 4H), 4.10-4.04 (m, 1H), 3.99-3.92 (m, 1H), 3.21 (d, J
= 6 Hz, 1H), 2.94-2.85 (m, 2H), 1.80-1.69 (m, 1H), 1.50 (s, 3H),
1.43 (s, 9H), 1.01-0.89 (m, 12H). XIV(f) ##STR00067##
C.sub.27H.sub.39F.sub.2N.sub.3O.sub.7 ((S)-1-{(S)-1-[(S)-1-
Benzyl-2-((R)- 2-methyl-oxiranyl)-2- oxo-ethylcarbamoyl]-3- methyl-
butylcarbamoyl}-2- difluoromethoxyethyl)- carbamic acid tert-butyl
ester off-white powder 82 .sup.1H NMR (300 MHz, CDCl.sub.3):
.delta. (ppm): 7.32-7.20 (m, 3H), 7.13 (dd, 2H), 6.82 (wd, 1H),
6.70 (wd, 1H), 6.51 (w t, 1H), 6.25 (t, J = 75 Hz, 1H), 5.26 (br s,
1H), 4.86 (td, 1H), 4.58 (td, 1H), 4.36 (m, 1H), 4.32- 4.08 (m,
2H), 4.01 (dd, 1H), 3.90 (dd, 1H), 3.24 (d, 1H), 3.12 (dd, 1H),
2.90 (d, 1H), 2.85 (dd, 1H), 1.81-1.70 (m, 1H), 1.50 (s, 3H), 1.43
(s, 9H), 1.03-0.88 (m, 6H). XIV(g) ##STR00068##
C.sub.27H.sub.39F.sub.2N.sub.3O.sub.7 ((S)-1-{(S)-1-[(S)-1-
Benzyl-2-((R)-2-methyl- oxiranyl)-2-oxo- ethylcarbamoyl]-2-
difluoromethoxy- ethylcarbamoyl}-3- methylbutyl)-carbamic acid
tert-butyl ester off-white powder 87 .sup.1H NMR (300 MHz,
CDCl.sub.3): .delta. (ppm): 7.34-7.21 (m, 3H), 7.12 (dd, 2H), 6.79
(wd, 1H), 6.73 (wd, 1H), 6.49 (wt, 1H), 6.24 (t, J = 75 Hz, 1H),
5.24 (br s, 1H), 4.83 (td, 1H), 4.55 (td, 1H), 4.38 (m, 1H), 4.35-
4.03 (m, 2H), 4.06 (dd, 1H), 3.88 (dd, 1H), 3.25 (d, 1H), 3.11 (dd,
1H), 2.89 (d, 1H), 2.83 (dd, 1H), 1.84-1.75 (m, 1H), 1.52 (s, 3H),
1.42 (s, 9H), 1.02-0.91 (m, 6H). XIV(h) ##STR00069##
C.sub.24H.sub.41F.sub.2N.sub.3O.sub.7 ((S)-1-{(S)-2-
Difluoromethoxy-1-[(S)-3- methyl-1-((R)-2-methyl- oxiranecarbonyl)-
butylcarbamoyl]- ethylcarbamoyl}-3- methylbutyl)-carbamic acid
tert-butyl ester off-white powder 67 .sup.1H NMR (300 MHz,
CDCl.sub.3): .delta. (ppm): 6.81 (wd, 1H) 6.72 (wd, 1H), 6.51 (wt,
1H), 6.25 (t, J = 75 Hz, 1H), 5.24 (br s, 1H), 4.83 (td, 1H), 4.55
(td, 1H), 4.38 (m, 1H), 4.35-4.03 (m, 2H), 4.06 (dd, 1H), 3.88 (dd,
1H), 3.25 (d, 1H), 3.11 (dd, 1H), 2.89 (d, 1H), 2.83 (dd, 1H),
1.80-1.71 (m, 2H), 1.52 (s, 3H), 1.42 (s, 9H), 1.05-0.93 (m,
12H).
TABLE-US-00004 TABLE 4 Compound Formula and Yield # Structure
Nomenclature Appearance (%) XV(b) ##STR00070##
C.sub.20H.sub.27F.sub.2N.sub.3O.sub.6 (free base)
(S)-2-Amino-N-{(S)-1- [(S)-1-benzyl- 2-((R)-2-methyl-oxiranyl)-
2-oxo-ethylcarbamoyl]-2- methoxyethyl}-3-
difluoromethoxypropionamide off-yellow solid 98 .sup.1H NMR (300
MHz, d.sub.6-DMSO): .delta. (ppm): 7.39-7.25 (m, 3H), 7.24 (dd,
2H), 6.87 (wd, 1H), 6.66 (wd, 1H), 6.55 (br s, 1H), 6.13 (w t, 1H),
6.02 (wt, 1H), 5.15 (br s, 1H), 4.81 (td, 1H), 4.49 (td, 1H), 4.32
(m, 1H), 4.93-3.31 (m, 2H), 3.99 (dd, 1H), 3.88 (dd, 1H), 3.55 (s,
3H). 3.21 (d, 1H), 3.12 (dd, 1H), 2.81 (d, 1H), 2.78 (dd, 1H), 1.43
(s 3H). XV(c) ##STR00071## C.sub.20H.sub.27F.sub.2N.sub.3O.sub.6
(free base) (S)-2-Amino-N-{(S)-1- [(S)-1-benzyl-
2-((R)-2-methyl-oxiranyl)- 2-oxo-ethylcarbamoyl]-2-
difluoromethoxyethyl}-3- methoxypropionamide off-yellow solid 85
.sup.1H NMR (300 MHz, d.sub.6-DMSO): .delta. (ppm): 7.33-7.27 (m,
3H), 7.25 (dd, 2H), 6.87 (d, 1H), 6.59 (wd, 1H), 6.57 (br s, 1H),
6.09 (w t, 1H), 6.11 (t, 1H), 5.21 (br s, 1H), 4.76 (td, 1H), 4.52
(td, 1H), 4.41 (m, 1H), 4.33-3.98 (m, 2H), 3.95 (dd, 1H), 3.89 (dd,
1H), 3.51 (s, 3H). 3.18 (d, 1H), 3.09 (dd, 1H), 2.83 (d, 1H), 2.76
(dd, 1H), 1.44 (s, 3H). XV(d) ##STR00072##
C.sub.17H.sub.27F.sub.4N.sub.3O.sub.6 (free base) (S)-2-Amino-3-
difluoromethoxy-N-{(S)-2- difluoromethoxy-1-[(S)-3-
methyl-1-((R)-2- methyloxiranecarbonyl)- butylcarbamoyl]-ethyl}-
propionamide sticky yellow powder 73 .sup.1H NMR (300 MHz,
d.sub.6-DMSO): .delta. (ppm): 6.97 (d, 1H), 6.69-6.65 (m, 1H),6.45
(br.s, 2H), 6.16 (t, J = 75 Hz, 1H), 6.13 (t, J = 75 Hz, 1H),
5.35-5.26 (m, 1H), 4.69-4.58 (m, 3H), 4.44-4.24 (m, 4H), 4.10-4.04
(m, 1H), 3.99-3.92 (m, 1H), 3.24 (d, J = 6 Hz, 1H), 2.91-2.87 (m,
2H), 1.72-1.69 (m, 1H), 1.49 (s, 3H), 0.94-0.91 (m, 6H). XV(e)
##STR00073## C.sub.19H.sub.33F.sub.2N.sub.3O.sub.5 (free base)
(S)-2-((S)-2-Amino-3- difluoromethoxy- propionylamino)-4-methyl-
pentanoic acid [(S)-3- methyl-1-((R)-2- methyloxiranecarbonyl)-
butyl]-amide white powder 65 .sup.1H NMR (300 MHz, d.sub.6-DMSO):
.delta. (ppm): 6.89 (d, 1H), 6.70-6.59 (m, 1H), 6.55 (br. s, 2H),
6.17, (t, J = 75 Hz, 1H), 5.35-5.24 (m, 1H), 4.71-4.63 (m, 3H),
4.51-4.53 (m, 4H), 4.13-4.01 (m, 1H), 3.97-3.91 (m, 1H), 3.23 (d,
1H), 2.98-2.81 (m, 2H), 1.78-1.65 (m, 1H), 1.48 (s, 3H), 0.98-0.83
(m, 12H). XV(f) ##STR00074## C.sub.22H.sub.31F.sub.2N.sub.3O.sub.5
(free base) (S)-2-((S)-2-Amino-3- difluoromethoxy-
propionylamino)-4-methyl- pentanoic acid [(S)-1- benzyl-2-((R)-2-
methyloxiranyl)-2-oxo- ethyl]-amide off-white powder 62 .sup.1H NMR
(300 MHz, d.sub.6-DMSO): .delta. (ppm): 7.29-7.18 (m, 3H), 7.09
(dd, 2H), 6.74 (wd, 1H), 6.67 (br. s, 2H), 6.65 (d, 1H), 6.48 (t,
1H), 6.19 (t, J = 75 Hz, 1H), 5.21 (br s, 1H), 4.77 (td, 1H), 4.51
(td, 1H), 4.38- 4.34 (m, 1H), 4.19-4.05 (m, 2H), 3.99 (dd, 1H),
3.95-3.92 (m, 1H), 3.18 (d, 1H), 3.14-3.16 (m, 1H), 2.85 (d, 1H),
2.75-7.62 (m, 1H), 1.77-1.69 (m, 1H), 1.48 (s, 3H), 0.97-0.86 (m,
6H). XV(g) ##STR00075## C.sub.22H.sub.31F.sub.2N.sub.3O.sub.5 (free
base) (S)-2-Amino-4-methyl- pentanoic acid {(S)-1-
[(S)-1-benzyl-2-((R)-2- methyloxiranyl)-2-oxo- ethylcarbamoyl]-2-
difluoromethoxyethyl}- amide off-white powder 76 .sup.1H NMR (300
MHz, d.sub.6-DMSO): .delta. (ppm): 7.31-7.19 (m, 3H), 7.11 (dd,
2H), 6.76-6.68 (m, 1H), 6.52 (br. s, 2H), 6.49 (d, 1H), 6.45 (t,
1H), 6.21 (t, J = 75 Hz, 1H), 5.18 (br. s, 1H), 4.76-4.68 (m, 1H),
4.54-4.49 (m, 1H), 4.29-4.25 (m, 1H), 4.15-4.01 (m, 2H), 3.98-3.91
(m, 1H), 3.87-3.82 (m, 1H), 3.15 (d, 1H), 3.15-3.11 (m, 1H), 2.76
(d, 1H), 2.77-7.60 (m, 1H), 1.89-1.78 (m, 1H), 1.47 (s, 3H),
0.98-0.87 (m, 6H). XV(h) ##STR00076##
C.sub.19H.sub.33F.sub.2N.sub.3O.sub.5 (S)-2-Amino-4-methyl-
pentanoic acid {(S)-2- difluoromethoxy-1-[(S)-3- methyl-1-((R)-2-
methyloxiranecarbonyl)- butylcarbamoyl]-ethyl}- amide off-white
powder 89 .sup.1H NMR (300 MHz, d.sub.6-DMSO): .delta. (ppm): 6.92
(wd, 1H), 6.72 (wd, 1H), 6.55-6.50 (m, 1H), 6.15 (t, J = 75 Hz,
1H), 5.09 (br. s, 1H), 4.81-4.78 (m, 1H), 4.57-4.52 (m, 1H),
4.28-4.19 (m, 1H), 4.17-4.02 (m, 2H), 3.99-3.87 (m, 1H), 3.73 (dd,
1H), 3.17 (d, 1H), 3.07 (dd, 1H), 2.75 (d, 1H), 2.81-2.77 (m, 1H),
1.73-1.69 (m, 2H), 1.49 (s, 3H), 0.98-0.91 (m, 12H).
TABLE-US-00005 TABLE 5 Example Yield # Structure Nomenclature
Appearance (%) 2 ##STR00077## 2-Methyl-thiazole-5- carboxylic
acid-((S)-1- {(S)-1-[(S)-1-benzyl-2- ((R)-2-methyl-oxiranyl)-
2-oxo-ethylcarbamoyl]- 2-methoxy- ethylcarbamoyl}-2-
difluoromethoxyethyl)- amide white yellowish powder 73 .sup.1H NMR
(300 MHz, CDCl.sub.3): .delta. (ppm): 8.06 (s, 1H), 7.27-7.24 (m,
3H), 7.18 (dd, 2H), 6.79 (d, 1H), 6.77 (d, 1H), 6.64 (d, 1H), 6.13
(wt, 1H), 4.87-4.67 (m, 2H), 4.58 (td, 1H), 4.32 (dd, 1H), 4.27
(dd, 1H), 4.02 (td, 1H), 3.91 (dd, 1H), 3.30 (s, 3H), 3.27 (d, 1H),
3.28 (dd, 1H), 3.16 (d, 1H), 2.88 (dd, 1H), 2.83 (dd, 1H), 2.74 (s,
3H), 1.46 (s, 3H). 3 ##STR00078## 2-Methyl-thiazole-5- carboxylic
acid ((S)-1-{(S)- 1-[(S)-1-benzyl-2-((R)-2- methyl-oxiranyl)-2-oxo-
ethylcarbamoyl]-2- difluoromethoxyethyl- carbamoyl}-
2-methoxyethyl)- amide off-white powder 65 .sup.1H NMR (300 MHz,
CDCl.sub.3): .delta. (ppm): 8.07 (s, 1H), 7.28-7.26 (m, 3H), 7.15
(dd, 2H), 6.81 (d, 1H), 6.82 (d, 1H), 6.71 (d, 1H), 6.21 (wt, 1H),
4.91-4.82 (m, 2H), 4.62 (td, 1H), 4.38 (dd, 1H), 4.31 (dd, 1H),
4.05 (td, 1H), 3.99 (dd, 1H), 3.29 (s, 3H), 3.27 (d, 1H), 3.25 (dd,
1H), 3.15 (d, 1H), 2.91 (dd, 1H), 2.79 (dd, 1H), 2.71 (s, 3H), 1.49
(s, 3H). 4 ##STR00079## 2-Methyl-oxazole-5- carboxylic acid
((S)-1-{(S)- 1-[(S)-1-benzyl-2-((R)-2- methyl-oxiranyl)-2-oxo-
ethylcarbamoyl]-2- difluoromethoxyethyl- carbamoyl}-2-
difluoromethoxyethyl)- amide off-white powder 45 .sup.1H NMR (300
MHz, CDCl.sub.3): .delta. (ppm): 8.05 (s, 1H), 7.30-7.27 (m, 3H),
7.18 (dd, 2H), 6.79 (d, 1H), 6.76 (d, 1H), 6.63 (d, 1H), 6.16 (wt,
1H), 6.10 (wt, 1H), 4.88-4.61 (m, 2H), 4.58 (td, 1H), 4.31 (dd,
1H), 4.22 (dd, 1H), 4.01 (td, 1H), 3.89 (dd, 1H), 3.26 (d, 1H),
3.24 (dd, 1H), 3.14 (d, 1H), 2.85 (dd, 1H), 2.85 (dd, 1H), 2.68 (s,
3H), 1.43 (s, 3H). 5 ##STR00080## 3-Methyl-isoxazole-5- carboxylic
acid ((S)-1-{(S)- 1-[(S)-1-benzyl-2-((R)-2- methyl-oxiranyl)-2-oxo-
ethylcarbamoyl]-2- difluoromethoxyethyl- carbamoyl}-2-
difluoromethoxyethyl)- amide white solid 75 .sup.1H NMR (300 MHz,
CDCl.sub.3): .delta. (ppm): 7.87 (s, 1H), 7.31-7.28 (m, 3H), 7.18
(dd, 2H), 6.79 (d, 1H), 6.76 (d, 1H), 6.63 (d, 1H), 6.14 (wt, 1H),
6.14 (wt, 1H), 4.78-4.55 (m, 2H), 4.55 (td, 1H), 4.29 (dd, 1H),
4.19 (dd, 1H), 4.02 (td, 1H), 3.91 (dd, 1H), 3.24 (d, 1H), 3.24
(dd, 1H), 3.14 (d, 1H), 2.85 (dd, 1H), 2.78 (dd, 1H), 2.45 (s, 3H),
1.45 (s, 3H). 6 ##STR00081## Thiazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl- 2-((R)-2-methyloxiranyl)-2-
oxoethylcarbamoyl]-2- difluoromethoxyethyl carbamoyl}-2-
difluoromethoxyethyl)amide off-white yellowish powder 63 .sup.1H
NMR (300 MHz, CDCl.sub.3): .delta. (ppm): 9.35 (s, 1H), 7.31-7.27
(m, 4H), 7.18 (dd, 2H), 6.92 (d, 1H), 6.79 (d, 1H), 6.66 (d, 1H),
6.21 (wt, 1H), 6.16 (wt, 1H), 4.92-4.71 (m, 2H), 4.68 (td, 1H),
4.28 (dd, 1H), 4.19 (dd, 1H), 4.02 (td, 1H), 3.95 (dd, 1H), 3.28
(d, 1H), 3.25 (dd, 1H), 3.13 (d, 1H), 2.88 (dd, 1H), 2.77 (dd, 1H),
1.43 (s, 3H). 7 ##STR00082## Oxazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl- 2-((R)-2-methyloxiranyl)-2-
oxo-ethylcarbamoyl]-2- difluoromethoxy- ethylcarbamoyl}-2-
difluoromethoxyethyl)- amide off-white powder 43 .sup.1H NMR (300
MHz, CDCl.sub.3): .delta. (ppm): 8.89 (s, 1H), 7.38-7.25 (m, 4H),
7.15 (dd, 2H), 6.88 (d, 1H), 6.77 (d, 1H), 6.63 (d, 1H), 6.18 (wt,
1H), 6.13 (wt, 1H), 4.83-4.69 (m, 2H), 4.59 (td, 1H), 4.28 (dd,
1H), 4.23 (dd, 1H), 4.05 (td, 1H), 3.98 (dd, 1H), 3.31 (d, 1H),
3.28 (dd, IH), 3.09 (d, 1H), 2.95 (dd, 1H), 2.89 (dd, 1H), 1.41 (s,
3H). 8 ##STR00083## 5-Methyl-thiophene-2- carboxylic acid ((S)-1-
{(S)-1-[(S)-1-benzyl-2- ((R)-2-methyl-oxiranyl)-2-
oxo-ethylcarbamoyl]-2- difluoro- methoxyethylcarbamoyl}-
2-difluoromethoxyethyl)- amide yellow powder 36 .sup.1H NMR (300
MHz, CDCl.sub.3): .delta. (ppm): 7.95 (s, 1H), 7.76 (s, 1H), 7.36-
7.28 (m, 3H), 7.21 (dd, 2H), 6.90 (d, 1H), 6.81 (d, 1H), 6.72 (d,
1H), 6.21 (wt, 1H), 6.22 (wt, 1H), 4.89-4.68 (m, 2H), 4.63 (td,
1H), 4.25 (dd, 1H), 4.14 (dd, 1H), 4.04 (td, 1H), 3.87 (dd, 1H),
3.33 (d, 1H), 3.28 (dd, 1H), 3.18 (d, 1H), 2.81 (dd, 1H), 2.77 (dd,
1H), 2.51 (s, 3H). 1.41 (s, 3H). 9 ##STR00084##
5-Methyl-furan-2-carboxylic acid ((S)-1-{(S)-1-[(S)-1-
benzyl-2-((R)-2- methyloxiranyl)-2-oxo- ethylcarbamoyl]-2-
difluoromethoxyethylcarbamoyl}- 2-difluoromethoxyethyl)- amide
off-white powder 69 .sup.1H NMR (300 MHz, CDCl.sub.3): .delta.
(ppm): 7.73 (s, 1H), 7.74 (s, 1H), 7.37- 7.25 (m, 3H), 7.18 (dd,
2H), 6.87 (d, 1H), 6.78 (d, 1H), 6.69 (d, 1H), 6.22 (wt, 1H), 6.19
(wt, 1H), 4.93-4.72 (m, 2H), 4.71 (td, 1H), 4.29 (dd, 1H), 4.18
(dd, 1H), 4.03 (td, 1H), 3.96 (dd, 1H), 3.41 (d, 1H), 3.34 (dd,
1H), 3.21 (d, 1H), 2.99 (dd, 1H), 2.75 (dd, 1H), 2.46 (s, 3H). 1.42
(s, 3H). 10 ##STR00085## Thiophene-2-carboxylic acid ((S)-1-
{(S)-1-[(S)-1-benzyl-2-((R)-2- methyloxiranyl)-2-oxo
ethylcarbamoyl]-2- difluoromethoxy- ethylcarbamoyl}-2-
difluoromethoxyethyl)-amide off-white powder 74 .sup.1H NMR (300
MHz, CDCl.sub.3): .delta. (ppm): 7.95 (d, 1H), 7.77-158 (m, 2H),
7.38-7.26 (m, 3H), 7.19 (dd, 2H), 6.88 (d, 1H), 6.79 (d, 1H), 6.68
(d, 1H), 6.18 (wt, 1H), 6.19 (wt, 1H), 4.91-4.72 (m, 2H), 4.58 (td,
1H), 4.17 (dd, 1H), 4.11 (dd, 1H), 4.01 (td, 1H), 3.91 (dd, 1H),
3.36 (d, 1H), 3.22 (dd, 1H), 3.13 (d, 1H), 2.91 (dd, 1H), 2.68 (dd,
1H), 1.45 (s, 3H). 11 ##STR00086## N-[(1S)-2-[[(1S)-2-[[(1S)-1-
benzyl-2-[(2R)-2-methyloxiran- 2-yl]-2-oxo-ethyl]amino]-
1(difluoromethoxymethyl)-2- oxo-ethyl]amino]-1-
(difluoromethoxymethyl)-2- oxo-ethyl]-1H-1,2,4-triazole-5-
carboxamide white powder 6 .sup.1H NMR (300 MHz, d.sub.6-DMSO):
.delta. (ppm): 8.58 (d, J = 4 HZ, 1H), 8.49 (d, J = 8 Hz, 1H),
7.29-7.16 (m, 6H), 6.61 (d, J = 76 Hz, 1H), 6.60 (d, J = 76 Hz,
1H), 4.83-4.76 (m, 1H), 4.63-4.54 (m, 2H), 4.14-4.08 (m, 2H),
3.99-3.85 (m, 2H), 3.18 (d, J = 6 Hz, 1H), 2.99-2.91 (m, 2H),
2.73-2.64 (m, 1H), 1.34 (s, 3H). 12 ##STR00087## N-[(1S)-1-
(difluoromethoxymethyl)-2- [[(1S)-1- (difluoromethoxymethyl)-2-
[[(1S)-3-methyl-1-[(2R)-2- methyloxirane-2-carbonyl]butyl]-
amino]-2-oxo-ethyl]amino]-2- oxo-ethyl]-2-methylthiazole-5-
carboxamide white powder 75 .sup.1H NMR (300 MHz, d.sub.6-DMSO):
.delta. (ppm): 8.88 (d, J = 4 Hz, 1H), 8.50 (d, J = 4 Hz, 1H),
8.34-8.28 (m, 2H), 6.65 (t, J = 76 Hz, 1H), 6.61 (t, J = 76 Hz,
1H), 4.83-4.74 (m, 1H), 4.58-4.49 (m, 1H), 4.40-4.31 (m, 1H), 4.15-
4.08 (m, 1H), 4.05-3.88 (m, 3H), 3.12 (d, J = 4 Hz, 1H), 2.98 (d, J
= 4 Hz, 1H), 2.65 (s, 3H), 1.67-1.55 (m, 1H), 1.38 (s, 3H),
1.35-1.22 (m, 2H), 0.86 (d, J = 6 Hz, 3H), 0.81 (d, J = 6 Hz, 3H).
13 ##STR00088## Pyridine-2-carboxylic acid
((S)-1-{(S)-1-[(S)-1-benzyl-2- ((R)-2-methyloxiranyl)-2-oxo-
ethylcarbamoyl]-2- difluoromethoxy- ethylcarbamoyl}-2-
difluoromethoxyethyl)-amide white powder 86 .sup.1H NMR (300 MHz,
d.sub.6-DMSO): .delta. (ppm): 8.39 (d, 1H), 7.45-7.35 (m, 2H),
7.31-7.14 (m, 6H), 6.59 (wt, J = 75 Hz, 1H), 6.57 (wt, J = 76 Hz,
1H), 4.85-4.78 (m, 1H), 4.71-4.56 (m, 2H), 4.12-4.05 (m, 2H),
4.01-3.87 (m, 2H), 3.19-3.14 (m, 1H), 3.02-2.94 (m, 2H), 2.68-2.58
(m, 1H), 1.36 (s, 3H). 14 ##STR00089##
N-((S)-1-{(S)-1-[(S)-1-Benzyl- 2-((R)-2-methyl-oxiranyl)-2-
oxo-ethylcarbamoyl]-2- difluoromethoxyethylcarbamoyl}-
2-difluoromethoxyethyl)- nicotinamide white powder 77 .sup.1H NMR
(300 MHz, d.sub.6-DMSO): .delta. (ppm): 8.33 (s, 1H), 8.28 (dd,
1H), 7.38-7.29 (m, 2H), 7.29-7.17 (m, 5H), 6.62 (wt, J = 75 Hz,
1H), 6.59 (wt, J = 76 Hz, 1H), 4.78-4.67 (m, 1H), 4.83-4.59 (m,
2H), 4.15-4.03 (m, 2H), 3.98-3.83 (m, 2H), 3.21-3.15 (m, 1H),
2.99-2.91 (m, 2H), 2.70-2.56 (m, 1H), 1.35 (s, 3H). 15 ##STR00090##
Pyridine-2-carboxylic acid ((S)-1-{(S)-1-[(S)-1-benzyl-2-
((R)-2-methyloxiranyl)-2-oxo- ethylcarbamoyl]-2- difluoromethoxy-
ethylcarbamoyl}-2- difluoromethoxy-ethyl)-amide white yellowish
powder 61 .sup.1H NMR (300 MHz, d.sub.6-DMSO): .delta. (ppm): 8.65
(s, 1H), 8.41-8.35 (m, 2H), 7.27-7.16 (m, 5H), 6.61 (wt, J = 75 Hz,
1H), 6.58 (wt, J = 76 Hz, 1H), 4.81-4.69 (m, 1H), 4.82-4.57 (m,
2H), 4.21-4.04 (m, 2H), 3.99-3.79 (m, 2H), 3.25-3.18 (m, 1H),
2.99-2.91 (m, 2H), 2.69-2.54 (m, 1H), 1.37 (s, 3H). 16 ##STR00091##
N-[(1S)-2-[[(1S)-2-[[(1S)-1- benzyl-2-[(2R)-2-methyloxiran-
2-yl]-2-oxo-ethyl]amino]-1- (difluoromethoxymethyl)-2-oxo-
ethyl]amino]-1- (difluoromethoxymethyl)-2-oxo-
ethyl]-pyrimidine-2-carboxamide white yellowish powder 56 .sup.1H
NMR (300 MHz, d.sub.6-DMSO): .delta. (ppm): 8.55-7.58 (m, 2H),
7.55-745 (m, 1H), 7.29-7.18 (m, 5H), 6.59 (wt, J = 75 Hz, 1H), 6.57
(wt, J = 76 Hz, 1H), 4.78-4.65 (m, 1H), 4.80-4.61 (m, 2H),
4.17-4.01 (m, 2H), 4.02- 3.85 (m, 2H), 3.23-3.17 (m, 1H), 2.98-2.87
(m, 2H), 2.70-2.55 (m, 1H), 1.35 (s, 3H). 17 ##STR00092##
[1,2,4]Triazine-3-carboxylic acid((S)-1-{(S)-1-[(S)-1-
benzyl-2-((R)-2-methyl- oxiranyl)-2-oxo- ethylcarbamoyl]-2-
difluoromethoxyethylcarbamoyl}- 2-difluoromethoxy-ethyl)- amide
white solid 51 .sup.1H NMR (300 MHz, d.sub.6-DMSO): .delta. (ppm):
9.61 (dd, 1H), 932 (dd, 1H), 7.32-7.15 (m, 5H), 6.60 (wt, J = 75
Hz, 1H), 6.58 (wt, J = 76 Hz, 1H), 4.71-4.61 (m, 1H), 4.76-4.59 (m,
2H), 4.19-3.99 (m, 2H), 4.03-3.88 (m, 2H), 3.25-3.16 (m, 1H),
2.97-2.85 (m, 2H), 2.68-2.53 (m, 1H), 1.37 (s, 3H). 18 ##STR00093##
Pyrimidine-4-carboxylic acid ((S)-1-{(S)-1-[(S)-1-benzyl-2-
((R)-2-methyloxiranyl)-2-oxo- ethylcarbamoyl]-2- difluoromethoxy-
ethylcarbamoyl}-2- difluoromethoxyethyl)-amide white solid 39
.sup.1H NMR (300 MHz, d.sub.6-DMSO): .delta. (ppm): 9.73 (s, 1H),
7.93 (dd, 1H), 7.68 (dd, 1H), 7.32-7.15 (m, 5H), 6.60 (wt, J = 75
Hz, 1H), 6.58 (wt, J = 76 Hz, 1H), 4.75-4.61 (m, 1H), 4.83-4.59 (m,
2H), 4.21-4.03 (m, 2H), 4.01-3.84 (m, 2H), 3.19-3.13 (m, 1H),
3.01-2.89 (m, 2H), 2.67-2.51 (m, 1H), 1.37 (s, 3H). 19 ##STR00094##
2-Methyl-thiazole-5- carboxylic acid ((S)-1-{(S)-1-
[(S)-1-benzyl-2-((R)-2-methyl- oxiranyl)-2-oxo- ethylcarbamoyl]-3-
methylbutylcarbamoyl}-2- difluoromethoxyethyl)-amide white
yellowish solid 48 .sup.1H NMR (300 MHz, d.sub.6-DMSO): .delta.
(ppm): 8.77 (s, 1H), 7.31-7.16 (m, 3H), 7.11 (dd, 2H), 6.71 (wd,
1H), 6.63 (d, 1H), 6.41 (t, 1H), 6.17 (t, J = 75 Hz, 1H), 5.21 (br.
s, 1H), 4.81-4.78 (m, 1H), 4.53 (td, 1H), 4.41-4.37 (m, 1H),
4.23-4.05 (m, 2H), 4.01 (dd, 1H), 3.97-3.90 (m, 1H), 3.25 (s, 3H),
3.21 (d, 1H), 3.16-3.13 (m, 1H), 2.79 (d, 1H), 2.81-7.65 (m, 1H),
1.79-1.68 (m, 1H), 1.47 (s, 3H), 0.97-0.89 (m, 6H). 20 ##STR00095##
2-Methyl-thiazole-5-carboxylic acid ((S)-2-difluoromethoxy-1-
{(S)-3-methyl-1-[(S)-3-methyl- 1-((R)-2- methyloxiranecarbonyl)-
butylcarbamoyl]- butylcarbamoyl}-ethyl)-amide white yellowish solid
67 .sup.1H NMR (300 MHz, d.sub.6-DMSO): .delta. (ppm): 8.65 (s,
1H), 6.98 (wd, 1H), 6.83 (wd, 1H), 6.61-6.53 (m, 1H), 6.17 (t, J =
75 Hz, 1H), 6.07 (br. s, 1H), 4.79-4.68 (m, 1H), 4.61-4.55 (m, 1H),
4.26-4.15 (m, 1H), 4.13-4.03 (m, 2H), 3.98-3.86 (m, 1H), 3.81 (dd,
1H), 3.21 (s, 3H) 3.27 (d, 1H), 3.12 (dd, 1H), 2.81 (d, 1H),
2.78-2.75 (m, 1H), 1.71-1.67 (m, 2H), 1.47 (s, 3H), 0.97-0.89 (m,
12H). 21 ##STR00096## 2-Methyl-thiazole-5-carboxylic acid
((S)-1-{(S)-2-difluoromethoxy- 1-[(S)-3-methyl-1-((R)-2-
methyloxiranecarbonyl)- butylcarbamoyl]- ethylcarbamoyl}-3-
methylbutyl)-amide white yellowish solid 34 .sup.1H NMR (300 MHz,
d.sub.6-DMSO): .delta. (ppm): 8.61 (s, 1H), 6.81-6.78 (m, 2H),
6.68-6.61 (m, 1H), 6.16 (t, J = 75 Hz, 1H), 5.99 (br. s, 1H),
4.79-4.59
(m, 2H), 4.26-4.06 (m, 3H), 3.99-3.89 (m, 1H), 3.76 (dd, 1H), 3.27
(d, 1H), 3.18 (s, 3H), 3.15 (dd, 1H), 2.93 (d, 1H), 2.75-2.71 (m,
1H), 1.681.64 (m, 2H), 1.48 (s, 3H), 0.98-0.90 (m, 12H). 22
##STR00097## 2-Methyl-thiazole-5-carboxylic acid
((S)-1-{(S)-1-[(S)-1- benzyl-2-((R)-2-methyl- oxiranyl)-2-oxo-
ethylcarbamoyl]-2- difluoromethoxyethylcarbamoyl}-
3-methylbutyl)-amide white yellowish solid 34 .sup.1H NMR (300 MHz,
d.sub.6-DMSO): .delta. (ppm): 8.65 (s, 1H), 7.28-7.17 (m, 3H), 7.09
(dd, 2H), 7.01-6.71 (m, 1H), 6.65 (d, 1H), 6.38 (t, 1H), 6.15 (t, J
= 75 Hz, 1H), 5.98 (br. s, 1H), 4.75-4.67 (m, 1H), 4.45 (td, 1H),
4.45-4.36 (m, 1H), 4.30-4.05 (m, 2H), 3.98 (dd, 1H), 3.95-3.88 (m,
1H), 3.19 (s, 3H), 3.17 (d, 1H), 3.15-3.08 (m, 1H), 2.81 (d, 1H),
2.77-7.60 (m, 1H), 1.81-1.72 (m, 1H), 1.46 (s, 3H), 0.98-0.87 (m,
6H). 23 ##STR00098## 2-Methyl-thiazole-5-carboxylic acid
((S)-1-{(S)-2- difluoromethoxy-1-[(S)-3- methyl-1-((R)-2-methyl-
oxiranecarbonyl)- butylcarbamoyl]-ethylcarbamoyl}-
2-phenylethyl)-amide yellow solid 28 .sup.1H NMR (300 MHz,
d.sub.6-DMSO): .delta. (ppm): 8.69 (s, 1H), 7.32-7.16 (m, 3H), 7.13
(dd, 2H), 7.11-6.91 (m, 1H), 6.72 (d, 1H), 6.48 (t, 1H), 6.17 (t, J
= 75 Hz, 1H), 6.08 (br. s, 1H), 4.75-4.67 (m, 1H), 4.45 (td, 1H),
4.45-4.36 (m, 1H), 4.30-4.05 (m, 2H), 3.98 (dd, 1H), 4.01-3.89 (m,
1H), 3.21 (s, 3H), 3.19 (d, 1H), 3.17-3.11 (m, 1H), 2.78 (d, 1H),
2.76-7.62 (m, 1H), 1.85-1.77 (m, 1H), 1.45 (s, 3H), 0.97-0.86 (m,
6H). 24 ##STR00099## 2-Methyl-thiazole-5- carboxylic acid
((S)-1-{(S)-1- [(S)-1-benzyl-2-((R)-2-methyl- oxiranyl)-2-oxo-
ethy!carbamoyl]-2- difluoromethoxyethylcarbamoyl}-
2-phenylethyl)-amide yellow pale solid 49 .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta. (ppm): .delta. (ppm): 8.45 (s, 1H), 7.35-7.07
(m, 10H), 6.81-6.76 (m, 1H), 6.62-6.57 (m, 1H), 6.19 (t, J = 72 Hz,
1H), 4.78-4.67 m, 1H), 4.63-4.52 (m, 2H), 4.28-4.19 (m, 1H),
4.03-3.93 (m, 1 H), 3.55 (br. s, 2H), .53-2.39 (m, 4H), 1.46 (s,
3H).
TABLE-US-00006 TABLE 6 Compound Yield # Structure Nomenclature
Appearance (%) XVII(b) ##STR00100## benzyl (2S)-3-
(difluoromethoxy)- 2-[[(2S)-3- (difluoromethoxy)- 2-[(2-
morpholinoacetyl)amino] propanoyl]amino]propanoate white solid 84
.sup.1H NMR (400 MHz, CDCl.sub.3): .delta. (ppm): 7.89 (m, 1H),
7.41-7.30 (m 5H), 7.04 (m, 1H), 6.22 (t, J = 74 Hz, 1H), 6.13(t, J
= 74 Hz, 1H), 5.22 (s, 2H), 4.84-4.78 (m, 1H), 4.77-4.72 (m, 1H),
4.32-4.27 (m, 2H), 4.21- 4.15 (m, 1H), 4.07-4.00 (m, 1H), 3.76-3.69
(m, 4H), 3.07 (s, 2H), 2.68- 2.52 (m, 4H). XVII(c) ##STR00101##
benzyl(2S)-2- [[(2S)-3-(difluoro- methoxy)-2-[(2-
morpholinoacetyl)- amino]- propanoyl]amino]- 4- methylpentanoate
off-white powder 89 .sup.1H NMR (400 MHz, CDCl.sub.3): .delta.
(ppm): 7.85 (m, 1H), 7.40-7.30 (m, 5H), 6.66 (m, 1H), 6.20 (t, J =
74 Hz, 1H), 5.20-5.12 (m, 2H), 4.72-4.58 (m, 2H), 4.26-4.22 (m,
1H), 4.03-3.97 (m, 1H), 3.75-3.67 (m, 4H), 3.06 (s, 2H), 2.55-2.50
(m, 4H), 1.72-1.62 (m, 1H), 0.96-0.85 (m, 6H). XVII(d) ##STR00102##
benzyl (2S)- 3(difluoromethoxy)-2- [[(2S)-2-[(2- morpholinoacetyl)
amino]-3-phenyl- propanoyl]amino]- propanoate off-white powder 76
.sup.1H NMR (400 MHz, CDCl.sub.3): .delta. (ppm): 7.81 (m, 1H),
7.41-6.95 (m 10 H), 6.17 (t, J = 74 Hz, 1H), 5.21-5.13 (m, 2H),
4.90-4.83 (m, 1H), 4.72-4.64 (m, 1H), 4.31-4.25 (m, 1H), 401-3.93
(m, 1H), 3.73-3.61 (m, 4H), 3.91- 2.49 (m, 4H), 2.51-2.43 (m, 4H).
XVII(e) ##STR00103## benzyl (2S)-3- (difluoromethoxy)-2-
[[(2S)-4-methyl-2-[(2- morpholinoacetyl)amino]
pentanoyl]amino]propanoate off-white powder 87
TABLE-US-00007 TABLE 7 Compound Yield # Structure Nomenclature
Appearance (%) XVIII(b) ##STR00104## (2S)-3- (difluoromethoxy)-2-
[[(2S)-3- (difluoromethoxy)-2- [(2-morpholino- acetyl)amino]
propanoyl]amino]- propanoic acid white solid 95 .sup.1H NMR (300
MHz, CDCl.sub.3): .delta. (ppm): 7.89 (m, 1H), 7.41-7.30 (m, 5H),
7.04 (m, 1H), 6.22 (t, J = 74 Hz, 1H), 6.13 (t, J = 74 Hz, 1H),
5.22 (s, 2H), 4.84-4.78 (m, 1H), 4.77-4.72 (m, 1H), 4.32-4.27 (m,
2H), 4.21- 4.15 (m, 1H), 4.07-4.00 (m, 1H), 3.76-3.69 (m, 4H), 3.07
(s, 2H), 2.68-2.52 (m, 4H). XVIII(c) ##STR00105## (2S)-2-[[(2S)-3-
(difluoromethoxy)- 2-[(2- morpholinoacetyl) amino]-
propanoyl]amino]- 4-methylpentanoic acid off-white sticky solid 99
.sup.1H NMR (300 MHz, CDCl.sub.3): .delta. (ppm): 7.85 (m, 1H),
7.40-7.30 (m, 5H), 6.66 (m, 1H), 6.20 (t, J = 74 Hz, 1H), 5.20-5.12
(m, 2H), 4.72-4.58 (m, 2H), 4.26-4.22 (m, 1H), 4.03-3.97 (m, 1H),
3.75-3.67 (m, 4H), 3.06 (s, 2H), 2.55-2.50 (m, 4H), 1.72-1.62 (m,
1H), 0.96-0.85 (m, 6H).
TABLE-US-00008 TABLE 8 Example Yield # Structure Nomenclature
Appearance (%) 26 ##STR00106## (S)-N-{(S)-1-[(S)-1-
Benzyl-2-((R)-2-methyl- oxiranyl)-2-oxo- ethylcarbamoyl]-2-
difluoromethoxyethyl}-3- difluoromethoxy-2-(2- morpholin-4-yl-
acetylamino)-propionamide white solid 53 .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta. (ppm): 7.94 (m, 1H), 7.32-7.12 (m, 5H), 6.87
(m, 1H), 6.70 (m, 1H), 6.22 (t, J = 74 Hz, 1H), 6.12 (t, J = 74 Hz,
1H), 4.88-4.78 (m, 1H), 4.66-4.55 (m, 1H), 4.32-4.26 (m, 1H),
4.23-4.14 (m, 1H), 4.08-4.02 (m, 1H), 3.92-3.88 (m, 1H), 3.77-3.68
(m, 4H), 3.28- 2.80 (m, 5H), 2.60-2.51 (m, 4H), 1.51 (s, 3H). 27
##STR00107## (2S)-N-[(1S)-1-benzyl-2- [(2R)-2-methyloxiran-2-
yl]-2-oxo-ethyl]-2-[[(2S)-3- (difluoromethoxy)-2-[(2-
morpholinoacetyl)amino] propanoyl]amino]-4- methylpentanamide
off-white solid 47 .sup.1H NMR (400 MHz, CDCl.sub.3): .delta.
(ppm): 7.85 (m, 1H), 7.38-7.12 (m, 5H), 6.82-6.78 (m, 1H), 6.24 (t,
J = 74 Hz, 1H), 6.13-6.11 (m, 1H), 4.71-4.58 (m, 3H), 4.42-4.35 (m,
1H), 4.15-4.05 (m, 1H), 3.83-3.75 (m, 4H), 3.33- 2.95 (m, 5H),
2.60-2.55 (m, 4H), 2.22 (s, 3H), 1.61-1.42 (m, 3H), 0.98- 0.87 (m,
6H). 28 ##STR00108## (2S)-2-[[(2S)-3- (difluoromethoxy)-2-[(2-
morpholino-acetyl)amino] propa-noyl]amino]-4-
methyl-N-[(1S)-3-methyl- 1-[(2R)-2-methyloxirane- 2-carbonyl]
butyl]pentanamide pale solid 71 .sup.1H NMR (400 MHz, CDCl.sub.3):
.delta. (ppm): 7.84 (m, 1H), 6.59 (m, 1H), 6.26 (t, J = 74 Hz, 1H),
6.22 (m, 1H), 4.70-4.64 (m, 1H), 4.61-4.55 (m, 1H), 4.43-4.36 (m,
1H), 4.29-4.02 (m, 1H), 3.75-3.71 (m, 4H), 3.28-3.25 (m, 1H), 3.07
(s, 2H), 2.91-2.87 (m, 1H), 2.57-2.53 (m, 4H), 1.68-1.50 (m, 4H),
1.51 (s, 3H), 1.31-1.22 (m, 2H), 0.97-0.89 (m, 12H). 29
##STR00109## (2S)-N-[(1S)-1-benzyl-2- [(2R)-2-methyloxiran-2-yl]-
2-oxo-ethyl]-2-[[(2S)-3- (difluoromethoxy)-2-[(2-
morpholinoacetyl)amino] propanoyl]amino]-3-(1-
methylcyclohexa-1,3,5- trien-1-yl)propanamide white powder 63
.sup.1H NMR (400 MHz, CDCl.sub.3): .delta. (ppm): .delta. 7.74 (m,
1H), 7.32-6.98 (m, 10H), 6.70 (m, 1H), 6.59 (m, 1H), 6.18 (t, J =
72 Hz, 1H), 4.81-4.68 m, 1H), 4.61-4.51 (m, 2H), 4.23 = 4.17 (m,
1H), 3.99-3.95 (m, 1H), 3.68 (br. s, 2H), 3.10-2.90 (m, 6H),
2.80-2.59 (m, 2H), 2.53-2.39 (m, 4H), 1.48 (s, 3H). 30 ##STR00110##
(S)-4-Methyl-2-(2- morpholin-4-yl-acetylamino)- pentanoic acid
{(S)-2- difluoromethoxy-1-[(S)-3- methyl-1-((R)-2-methyl-
oxiranecarbonyl)- butylcarbamoyl]-ethyl}- amide off-white solid 34
.sup.1H NMR (400 MHz, CDCl.sub.3): .delta. (ppm): 7.88-7.76 (m,
1H), 6.63-6.59 (m, 1H), 6.29 (t, J = 75 Hz, 1H), 6.34 (m, 1H),
4.81-4.75 (m, 1H), 4.70-4.63 (m, 1H), 4.47-4.4 (m, 1H), 4.32-4.05
(m, 1H), 3.81-3.74 (m, 4H), 3.36- 3.26 (m, 1H), 3.11 (s, 2H),
3.01-2.95 (m, 1H), 2.66-2.61 (m, 4H), 1.73- 1.65 (m, 4H), 1.52 (s,
3H), 1.42-1.36 (m, 2H), 0.99-0.91 (m, 12H). 31 ##STR00111##
(S)-4-Methyl-2-(2-morpholin-4- yl-acetylamino)-pentanoic acid
{(S)-1-[(S)-1-benzyl-2-((R)-2- methyl-oxiranyl)-
2-oxo-ethylcarbamoyl]-2- difluoro- methoxyethyl}-amide off-white
solid 72 .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. (ppm): 8.01 (m,
1H), 7.02 (m, 1H), 6.79 (m, 1H), 6.41 (t, J = 74 Hz, 1H), 6.35 (t,
J = 74 Hz, 1H), 4.74-4.68 (m, 3H), 4.88-4.24 (m, 2H), 4.14-4.08 (m,
1H), 3.97-3.89 (m, 1H), 3.76-3.68 (m, 4H), 3.22 (d, J = 4 Hz, 1H),
2.90 (d, J = 4 Hz, 1H), 2.58-2.53 (m, 4H), 1.63-1.50 (m, 2H), 1.51
(s, 3H), 1.36-1.26 (m, 1H), 0.96-0.91 (m, 6H). 32 ##STR00112##
(2S)-N-[(1S)-2-[[(1S)-1-benzyl-2- [(2R)-2-methyloxiran-2-yl]-2-
oxo-ethyl]amino]- 1(difluoromethoxymethyl)-2-oxo-
ethyl]-2-[(2-morpholino- acetyl)amino]-3-phenyl- propanamide
off-white powder 61 .sup.1H NMR (400 MHz, CDCl.sub.3): .delta.
(ppm): 7.82 (m, 1H), 7.42-7.01 (m, 10H), 6.77-6.68 (m, 1H),
6.65-6.58 (m, 1H), 6.18 (t, J = 74 Hz, 1H), 4.86-4.72 (m, 1H),
4.72-4.60 (m, 2H), 4.23-4.21-4.16 (m, 1H), 402-3.98 (m, 1H), 3.71
(br. s, 2H), 3.14-2.95 (m, 6H), 2.82-2.69 (m, 2H), 2.62-2.47 (m,
4H), 1.49 (s, 3H). 33 ##STR00113## (2S)-3-(difluoromethoxy)-N-
[(1S)-1- (difluoromethoxymethyl)-2- [[(1S)-3-methyl-1-[(2R)-2-
methyloxirane-2- carbonyl]butyl]amino]-2-oxo- ethyl]-2-[(2-
morpholinoacetyl)amino] propanamide off-white powder 71 NMR
(CDCl.sub.3, 400 MHz): .delta. (ppm): 7.93 (m, 1H), 6.95 (m, 1H),
6.58 (m, 1H), 6.29 (t, J = 74 Hz, 1H), 6.22 (t, J = 74 Hz, 1H),
4.74-4.68 (m, 3H), 4.88-4.24 (m, 2H), 4.14-4.08 (m, 1H), 3.97-3.89
(m, 1H), 3.76-3.68 (m, 4H), 3.22 (d, J = 4 Hz, 1H), 2.90 (d, J = 4
Hz, 1H), 2.58-2.53 (m, 4H), 1.63-1.50 (m, 2H), 1.51 (s, 3H),
1.36-1.26 (m, 1H), 0.96-0.91 (m, 6H).
TABLE-US-00009 TABLE 9 Cell line OCI-AML-2 KMS-11 Ex- IC.sub.50
(nM)* am- CT-L CT-L ple proteasome proteasome # Cell Viability
activity Cell Viability activity 1 108.7 .+-. 25.31 39.17 .+-.
2.121 30.90 .+-. 10.71 29.62 .+-. 3.69 25 55.60 .+-. 19.20 153.9
.+-. 14.81 39.55 .+-. 7.45 172.9 .+-. 14.15 27 400.6 .+-. 275.4
599.9 .+-. 57.47 249.6 .+-. 25.89 742.5 .+-. 83.71 28 131.6 .+-.
87.55 556.1 .+-. 122.9 100.3 .+-. 18.69 575.1 .+-. 62.72 26 223.2
.+-. 10.34 358.5 .+-. 176.8 99.67 .+-. 17.73 174.1 .+-. 434.5 33
140.2 .+-. 53.47 598.8 .+-. 77.65 46.71 .+-. 9.89 703.8 .+-. 173 29
75.86 .+-. 14.97 131.7 .+-. 10.38 60.4 .+-. 11.18 135.3 .+-. 12.3
*Unless otherwise indicated, all experiments have been performed
independently at least twice, and results are presented as the mean
.+-. standard deviation.
TABLE-US-00010 TABLE 10* Example # chymotrypsin-like (CT-L)
Trypsin-like (T-L) Caspase-like (C-L) (Dose) 0 30 min 24 hours 0 30
min 24 hrs 0 30 min 24 hrs Vehicle 100% 98% 101% 100% 100% 100%
100% 100% 100% 1 (50 mg) 100% 27% 22% 100% 100% 100% 100% 100% 100%
26 (60 mg) 100% 33% 27% 100% 100% 100% 100% 100% 100% *Data are
presented as mean residual activity (SEM) relative to vehicle
treated controls.
TABLE-US-00011 TABLE 11 Panel/Cell Line Log.sub.10GI.sub.50
Log.sub.10TGI.sub.50 Log.sub.10LC.sub.50 Leukemia CCRF-CEM -7.26
>-4.00 >-4.00 HL-60(TB) -6.56 >-4.00 >-4.00 K-562 -6.50
>-4.00 >-4.00 MOLT-4 -7.37 >-4.00 >-4.00 RPMI-8226
-7.40 -4.86 >-4.00 SR -7.30 >-4.00 >-4.00 Non-Small Cell
Lung Cancer A549/ATCC -6.41 >-4.00 >-4.00 EKVX -6.22
>-4.00 >-4.00 HOP-62 -6.41 >-4.00 >-4.00 HOP-92 -6.23
-5.40 -4.39 NCI-H226 -7.44 -6.84 -- NCI-H23 -6.85 -5.92 -5.27
NCI-H322M -5.54 -4.74 -4.23 NCI-H460 -6.48 -5.84 -5.12 NCI-H522
-6.86 -6.15 -4.15 Colon Cancer COLO 205 -6.22 -4.85 >-4.00
HCC-2998 -6.74 -6.45 -6.15 HCT-116 -7.32 -6.71 -6.17 HCT-15 -5.65
-4.93 -4.16 HT29 -7.29 -6.22 -4.32 KM12 -6.88 -6.49 -6.10 SW-620
-7.39 -5.95 -4.40 CNS Cancer SF-268 -6.98 -5.90 >-4.00 SF-295
-7.17 -6.24 -4.88 SF-539 -7.15 -6.44 -4.27 SNB-19 -6.49 >-4.00
>-4.00 SNB-75 -6.81 -6.24 >-4.00 U251 -6.55 -4.94 >-4.00
Melanoma LOX IMVI -6.96 -6.56 -6.16 MALME-3M -7.30 -6.38 >-4.00
M14 -6.86 -6.24 -4.35 MDA-MB-435 -7.43 -6.74 >-4.00 SK-MEL-28
-6.85 -6.12 -4.69 SK-MEL-5 -7.15 -6.34 -5.47 UACC-257 -6.87 -6.20
>-4.00 UACC-62 -6.68 -5.78 -4.95 Ovarian Cancer OVCAR-3 -7.46
-6.90 -6.30 OVCAR-4 -6.29 >-4.00 >-4.00 OVCAR-5 -6.18 -5.30
-4.17 OVCAR-8 -5.80 >-4.00 >-4.00 NCI/ADR-RES -5.11 -4.23
>-4.00 SK-OV-3 -6.06 >-4.00 >-4.00 Renal Cancer 786-0
-6.43 -4.62 >-4.00 A498 -8.00 -6.38 -4.88 ACHN -6.53 -4.81
>-4.00 CAKI-1 -6.40 -5.28 >-4.00 RXF 393 -6.79 -6.41 -6.02
SN12C -6.54 -5.29 -4.03 TK-10 -6.71 -6.06 -- UO-31 -5.84 -5.32
-4.59 Prostate Cancer PC-3 -6.48 -4.99 >-4.00 DU-145 -6.58 -5.64
-5.02 Breast Cancer MCF7 -7.35 -4.62 >-4.00 MDA-MB- -6.66 -6.01
-4.73 231/ATCC HS 578T -7.33 -5.44 >-4.00 BT-549 -7.49 -6.87
-5.74 T-47D -6.59 >-4.00 -- MDA-MB-468 -6.75 -6.32 --
TABLE-US-00012 TABLE 12 Panel/Cell Line Log.sub.10GI.sub.50
Log.sub.10TGI.sub.50 Log.sub.10LC.sub.50 Leukemia HL-60(TB) -7.45
-6.35 >-4.00 K-562 -7.41 >-4.00 >-4.00 MOLT-4 -7.69 -7.11
>-4.00 RPMI-8226 -7.52 -7.04 >-4.00 SR -7.71 >-4.00
>-4.00 Non-Small Cell Lung Cancer A549/ATCC -7.41 -6.13
>-4.00 HOP-62 -7.75 -7.42 -7.09 HOP-92 -7.65 -7.16 -6.35
NCI-H226 -7.77 -7.43 -7.10 NCI-H23 -7.62 -7.13 -5.64 NCI-H322M
-7.10 -6.33 >-4.00 NCI-H460 -7.21 -6.43 -4.13 NCI-H522 -7.67
-7.19 >-4.00 Colon Cancer COLO 205 -7.39 -6.76 -6.21 HCC-2998
-7.74 -7.47 -7.20 HCT-116 -7.70 -7.30 -6.55 HCT-15 -6.47 >-4.00
>-4.00 HT29 -7.45 -5.84 >-4.00 KM12 -7.72 -7.39 -7.06 SW-620
-7.41 -5.93 >-4.00 CNS Cancer SF-268 -7.60 -7.14 >-4.00
SF-295 -7.68 -7.26 -6.49 SF-539 -7.63 -7.26 -6.48 SNB-19 -7.23
>-4.00 >-4.00 SNB-75 -7.71 -7.24 -6.35 U251 -7.44 -6.75
>-4.00 Melanoma LOX IMVI -7.73 -7.42 -7.10 MALME-3M -7.51 -6.64
>-4.00 M14 -7.48 -6.71 >-4.00 MDA-MB-435 -7.75 -7.42 -7.10
SK-MEL-2 -7.54 -7.03 -5.46 SK-MEL-28 -7.38 -6.69 -4.82 SK-MEL-5
-7.60 -7.17 -6.49 UACC-257 -7.18 -6.32 >-4.00 UACC-62 -7.35
-6.63 -4.58 Ovarian Cancer IGROV1 -7.43 -6.38 >-4.00 OVCAR-3
-7.66 -7.33 -6.97 OVCAR-4 -7.50 -6.75 >-4.00 OVCAR-5 -7.36 -6.44
>-4.00 OVCAR-8 -7.32 -6.41 >-4.00 NCI/ADR-RES -5.82 >-4.00
>-4.00 SK-OV-3 -7.30 -6.60 -- Renal Cancer 786-0 -7.20 -6.49
-5.44 A498 -7.61 -7.03 -6.34 ACHN -7.01 >-4.00 >-4.00 CAKI-1
-6.72 -6.09 >-4.00 RXF 393 -7.64 -7.25 -6.70 SN12C -7.39 -6.67
-5.15 TK-10 -7.49 -6.07 >-4.00 UO-31 -6.62 -5.95 -5.17 Prostate
Cancer PC-3 -7.45 -6.80 -5.14 DU-145 -7.27 -5.32 >-4.00 Breast
Cancer MCF7 -7.53 -4.89 >-4.00 MDA-MB- -7.52 -6.80 -4.21
231/ATCC HS 578T -7.54 -6.24 >-4.00 BT-549 -7.73 -7.39 -7.05
T-47D -7.78 -7.46 -- MDA-MB-468 -7.54 -6.96 >-4.00
TABLE-US-00013 TABLE 13 Panel/Cell Line Log.sub.10GI.sub.50
Log.sub.10TGI.sub.50 Log.sub.10LC.sub.50 Leukemia HL-60(TB) -7.45
-6.35 >-4.00 K-562 -7.41 >-4.00 >-4.00 MOLT-4 -7.69 -7.11
>-4.00 RPMI-8226 -7.52 -7.04 >-4.00 SR -7.71 >-4.00
>-4.00 Non-Small Cell Lung Cancer A549/ATCC -7.41 -6.13
>-4.00 HOP-62 -7.75 -7.42 -7.09 HOP-92 -7.65 -7.16 -6.35
NCI-H226 -7.77 -7.43 -7.10 NCI-H23 -7.62 -7.13 -5.64 NCI-H322M
-7.10 -6.33 >-4.00 NCI-H460 -7.21 -6.43 -4.13 NCI-H522 -7.67
-7.19 >-4.00 Colon Cancer COLO 205 -7.39 -6.76 -6.21 HCC-2998
-7.74 -7.47 -7.20 HCT-116 -7.70 -7.30 -6.55 HCT-15 -6.47 >-4.00
>-4.00 HT29 -7.45 -5.84 >-4.00 KM12 -7.72 -7.39 -7.06 SW-620
-7.41 -5.93 >-4.00 CNS Cancer SF-268 -7.60 -7.14 >-4.00
SF-295 -7.68 -7.26 -6.49 SF-539 -7.63 -7.26 -6.48 SNB-19 -7.23
>-4.00 >-4.00 SNB-75 -7.71 -7.24 -6.35 U251 -7.44 -6.75
>-4.00 Melanoma LOX IMVI -7.73 -7.42 -7.10 MALME-3M -7.51 -6.64
>-4.00 M14 -7.48 -6.71 >-4.00 MDA-MB-435 -7.75 -7.42 -7.10
SK-MEL-2 -7.54 -7.03 -5.46 SK-MEL-28 -7.38 -6.69 -4.82 SK-MEL-5
-7.60 -7.17 -6.49 UACC-257 -7.18 -6.32 >-4.00 UACC-62 -7.35
-6.63 -4.58 Ovarian Cancer IGROV1 -7.43 -6.38 >-4.00 OVCAR-3
-7.66 -7.33 -6.97 OVCAR-4 -7.50 -6.75 >-4.00 OVCAR-5 -7.36 -6.44
>-4.00 OVCAR-8 -7.32 -6.41 >-4.00 NCI/ADR-RES -5.82 >-4.00
>-4.00 SK-OV-3 -7.30 -6.60 Renal Cancer 786-0 -7.20 -6.49 -5.44
A498 -7.61 -7.03 -6.34 ACHN -7.01 >-4.00 >-4.00 CAKI-1 -6.72
-6.09 >-4.00 RXF 393 -7.64 -7.25 -6.70 SN12C -7.39 -6.67 -5.15
TK-10 -7.49 -6.07 >-4.00 UO-31 -6.62 -5.95 -5.17 Prostate Cancer
PC-3 -7.45 -6.80 -5.14 DU-145 -7.27 -5.32 >-4.00 Breast Cancer
MCF7 -7.53 -4.89 >-4.00 MDA-MB- -7.52 -6.80 -4.21 231/ATCC HS
578T -7.54 -6.24 >-4.00 BT-549 -7.73 -7.39 -7.05 T-47D -7.78
-7.46 -- MDA-MB-468 -7.54 -6.96 >-4.00
TABLE-US-00014 TABLE 14 Panel/Cell Line Log.sub.10GI.sub.50
Log.sub.10TGI.sub.50 Log.sub.10LC.sub.50 Leukemia CCRF-CEM -7.62
>-4.00 >-4.00 K-562 -7.00 >-4.00 >-4.00 MOLT-4 -7.54
-7.05 >-4.00 RPMI-8226 -7.40 -6.43 >-4.00 SR -7.69 -4.36
>-4.00 Non-Small Cell Lung Cancer A549/ATCC -6.68 >-4.00
>-4.00 HOP-62 -7.31 -6.53 -4.32 HOP-92 -7.18 -6.55 -6.03
NCI-H226 -7.64 -7.24 -6.51 NCI-H23 -7.21 -5.55 >-4.00 NCI-H322M
-6.27 -5.52 -4.18 NCI-H460 -6.49 -5.87 >-4.00 NCI-H522 -6.90
-6.17 >-4.00 Colon Cancer COLO 205 -6.90 -6.32 -5.34 HCC-2998
-6.80 -6.50 -6.21 HCT-116 -7.46 -5.91 >-4.00 HCT-15 -6.20
>-4.00 >-4.00 HT29 -7.31 -4.77 >-4.00 KM12 -7.35 -6.77
-6.27 SW-620 -7.40 -4.92 >-4.00 CNS Cancer SF-268 -7.27 -6.50
>-4.00 SF-295 -7.19 -6.55 -5.94 SF-539 -7.39 -6.76 -4.49 SNB-19
-6.51 >-4.00 >-4.00 SNB-75 -6.83 -6.41 -5.96 U251 -6.98 -5.70
>-4.00 Melanoma LOX IMVI -7.50 -6.90 -6.35 MALME-3M -6.69 -6.15
>-4.00 M14 -7.21 -5.85 >-4.00 MDA-MB-435 -7.63 -7.17 --
SK-MEL-2 -7.19 -6.02 >-4.00 SK-MEL-28 -7.13 -6.34 >-4.00
SK-MEL-5 -7.22 -6.66 -6.20 UACC-257 -6.81 -6.00 >-4.00 UACC-62
-7.00 -6.37 >-4.00 Ovarian Cancer IGROV1 -6.67 >-4.00
>-4.00 OVCAR-3 -7.35 -6.71 -5.66 OVCAR-4 -6.98 >-4.00 --
OVCAR-5 -6.65 -5.58 >-4.00 OVCAR-8 -6.59 >-4.00 >-4.00
NCI/ADR-RES -5.58 >-4.00 >-4.00 SK-OV-3 -6.46 -5.78 >-4.00
Renal Cancer 786-0 -6.55 -5.96 -4.87 A498 -6.68 -6.11 -5.47 ACHN
-6.59 >-4.00 >-4.00 CAKI-1 -6.56 -5.94 >-4.00 RXF 393
-7.20 -6.70 -6.31 SN12C -6.92 -6.38 -4.39 TK-10 -6.55 -5.10
>-4.00 UO-31 -6.46 -5.71 -4.62 Prostate Cancer PC-3 -7.04 -6.40
>-4.00 DU-145 -7.00 -5.05 -4.09 Breast Cancer MCF7 -7.38
>-4.00 >-4.00 MDA-MB- -7.04 -6.27 -4.44 231/ATCC HS 578T
-7.07 >-4.00 >-4.00 BT-549 -7.26 -6.59 -4.94 T-47D -7.16
-6.62 -- MDA-MB-468 -6.67 -6.10 >-4.00
* * * * *