U.S. patent application number 12/169209 was filed with the patent office on 2009-01-22 for inhibitors of serine protease, particularly hcv ns3-ns4a protease.
Invention is credited to Govinda Rao Bhisetti, Luc J. Farmer, Robert B. Perni, Keith P. Wilson.
Application Number | 20090022688 12/169209 |
Document ID | / |
Family ID | 33302781 |
Filed Date | 2009-01-22 |
United States Patent
Application |
20090022688 |
Kind Code |
A1 |
Farmer; Luc J. ; et
al. |
January 22, 2009 |
INHIBITORS OF SERINE PROTEASE, PARTICULARLY HCV NS3-NS4A
PROTEASE
Abstract
The present invention relates to compounds of formula I:
##STR00001## or a pharmaceutically acceptable salt, or mixtures
thereof, that inhibit serine protease activity, particularly the
activity of hepatitis C virus NS3-NS4A protease. As such, they act
by interfering with the life cycle of the hepatitis C virus and are
useful as antiviral agents. The invention further relates to
pharmaceutically acceptable compositions comprising said compounds
either for ex vivo use or for administration to a patient suffering
from HCV infection and processes for preparing the compounds. The
invention also relates to methods of treating an HCV infection in a
patient by administering a pharmaceutical composition comprising a
compound of this invention.
Inventors: |
Farmer; Luc J.; (Foxboro,
MA) ; Perni; Robert B.; (Marlborough, MA) ;
Bhisetti; Govinda Rao; (Lexington, MA) ; Wilson;
Keith P.; (La Jolla, CA) |
Correspondence
Address: |
VERTEX PHARMACEUTICALS INC.
130 WAVERLY STREET
CAMBRIDGE
MA
02139-4242
US
|
Family ID: |
33302781 |
Appl. No.: |
12/169209 |
Filed: |
July 8, 2008 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10821793 |
Apr 9, 2004 |
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12169209 |
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60513765 |
Oct 23, 2003 |
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Current U.S.
Class: |
424/85.5 ;
424/85.6; 424/85.7; 514/235.8; 514/255.05; 544/120; 544/406 |
Current CPC
Class: |
C07K 5/0812 20130101;
A61K 38/00 20130101; A61P 43/00 20180101; C07D 403/12 20130101;
C07K 5/0808 20130101; C07K 5/0804 20130101; A61P 31/14 20180101;
A61P 31/12 20180101; C07D 417/12 20130101 |
Class at
Publication: |
424/85.5 ;
544/406; 514/255.05; 544/120; 514/235.8; 424/85.7; 424/85.6 |
International
Class: |
A61K 38/21 20060101
A61K038/21; C07D 403/12 20060101 C07D403/12; A61K 31/497 20060101
A61K031/497; C07D 413/12 20060101 C07D413/12; A61K 31/5377 20060101
A61K031/5377 |
Claims
1. A compound of formula I: ##STR00068## or a pharmaceutically
acceptable salt, or mixtures thereof, wherein: W is: ##STR00069##
wherein each R.sub.6 is independently: hydrogen-,
(C1-C12)-aliphatic-, (C6-C10)-aryl-,
(C6-C10)-aryl-(C1-C12)aliphatic-, (C3-C10)-cycloalkyl- or
cycloalkenyl-, [(C3-C10)-cycloalkyl- or
cycloalkenyl]-(C1-C12)-aliphatic-, (C3-C10)-heterocyclyl-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, (C5-C10)-heteroaryl-, or
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-, or wherein up to 3
aliphatic carbon atoms in each R.sub.6 may be optionally replaced
with S, --S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)-- in a
chemically stable arrangement; wherein R.sub.6 may be optionally
substituted with up to 3 J substituents; or two R.sub.6 groups,
together with the nitrogen atom to which they are bound, may
optionally form a 5- to 6-membered aromatic or a 3- to 7-membered
saturated or partially unsaturated ring system wherein up to 3 ring
atoms may be optionally replaced with N, NH, O, S, SO, and
SO.sub.2, wherein said ring system may be optionally fused to a
(C6-C10)aryl, (C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a
(C3-C10)heterocyclyl, wherein any ring has up to 3 substituents
selected independently from J; wherein each R.sub.8 is
independently --OR'; or the R.sub.8 groups together with the boron
atom, may optionally form a (C3-C10)-membered heterocyclic ring
wherein each R.sub.8 is independently --OR'; or the R.sub.8 groups
together with the boron atom, may optionally form a
(C3-C10)-membered heterocyclic ring having, in addition to the
boron, up to 3 ring atoms optionally replaced with N, NH, O, S, SO,
and SO.sub.2; J is halogen, --OR', --NO.sub.2, --CN, --CF.sub.3,
--OCF.sub.3, --R', oxo, thioxo, .dbd.N(R'), .dbd.N(OR'),
1,2-methylenedioxy, 1,2-ethylenedioxy, --N(R').sub.2, --SR',
--SOR', --SO.sub.2R', --SO.sub.2N(R').sub.2, --SO.sub.3R',
--C(O)R', --C(O)C(O)R', --C(O)C(O)OR', --C(O)C(O)NR',
--C(O)CH.sub.2C(O)R', --C(S)R', --C(S)OR', --C(O)OR', --OC(O)R',
--C(O)N(R').sub.2, --OC(O)N(R').sub.2, --C(S)N(R').sub.2,
--(CH.sub.2).sub.0-2NHC(O)R', --N(R')N(R')COR',
--N(R')N(R')C(O)OR', --N(R')N(R')CON(R').sub.2, --N(R')SO.sub.2R',
--N(R')SO.sub.2N(R').sub.2, --N(R')C(O)OR', --N(R')C(O)R',
--N(R')C(S)R', --N(R')C(O)N(R').sub.2, --N(R')C(S)N(R').sub.2,
--N(COR')COR', --N(OR')R', --C(.dbd.NH)N(R').sub.2, --C(O)N(OR')R',
--C(.dbd.NOR')R', --OP(O)(OR').sub.2, --P(O)(R').sub.2,
--P(O)(OR').sub.2, or --P(O)(H)(OR'); wherein; R' is independently
selected from: hydrogen-, (C1-C12)-aliphatic-, (C3-C10)-cycloalkyl-
or -cycloalkenyl-, [(C3-C10)-cycloalkyl or
-cycloalkenyl]-(C1-C12)-aliphatic-, (C6-C10)-aryl-,
(C6-C10)-aryl-(C1-C12)aliphatic-, (C3-C10)-heterocyclyl-,
(C3-C10)-heterocyclyl-(C1-C12)aliphatic-, (C5-C10)-heteroaryl-, and
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; wherein up to 5 atoms in
R' may be optionally and independently substituted with J; wherein
two R' groups bound to the same atom may optionally form a 5- to
6-membered aromatic or a 3- to 7-membered saturated or partially
unsaturated ring system wherein up to 3 ring atoms may be
optionally replaced with a heteroatom independently selected from
N, NH, O, S, SO, and SO.sub.2, wherein said ring system may be
optionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,
(C3-C10)cycloalkyl, or a (C3-C10)heterocyclyl, wherein any ring has
up to 3 substituents selected independently from J; R.sub.5 and
R.sub.5' are each independently hydrogen or (C1-C12)-aliphatic,
wherein any hydrogen may be optionally replaced with halogen;
wherein any terminal carbon atom of R.sub.5 may be optionally
substituted with sulfhydryl or hydroxy; or R.sub.5 is Ph or
--CH.sub.2Ph and R.sub.5' is H, wherein said Ph or --CH.sub.2Ph
group may be optionally substituted with up to 3 substituents
independently selected from J; or R.sub.5 and R.sub.5' together
with the atom to which they are bound may optionally form a 3- to
6-membered saturated or partially unsaturated ring system wherein
up to 2 ring atoms may be optionally replaced with N, NH, O, SO, or
SO.sub.2; wherein said ring system has up to 2 substituents
selected independently from J; R.sub.2, R.sub.4, and R.sub.7 are
each independently: hydrogen-, (C1-C12)-aliphatic-,
(C3-C10)-cycloalkyl-(C1-C12)-aliphatic-, or
(C6-C10)-aryl-(C1-C12)-aliphatic-; wherein up to two aliphatic
carbon atoms in each of R.sub.2, R.sub.4, and R.sub.7 may be
optionally replaced with S, --S(O)--, --S(O).sub.2--, --O--, --N--,
or --N(H)-- in a chemically stable arrangement; wherein each of
R.sub.2, R.sub.4, and R.sub.7 may be independently and optionally
substituted with up to 3 substituents independently selected from
J; R.sub.1 and R.sub.3 are each independently: (C1-C12)-aliphatic-,
(C3-C10)-cycloalkyl- or -cycloalkenyl-, [(C3-C10)-cycloalkyl- or
-cycloalkenyl]-(C1-C12)-aliphatic-,
(C6-C10)-aryl-(C1-C12)aliphatic-, or
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; wherein up to 3 aliphatic
carbon atoms in each of R.sub.1 and R.sub.3 may be optionally
replaced with S, --S(O)--, --S(O).sub.2--, --O--, --N--, or
--N(H)-- in a chemically stable arrangement; wherein each of
R.sub.1 and R.sub.3 may be independently and optionally substituted
with up to 3 substituents independently selected from J; R.sub.9,
R.sub.9', R.sub.10, and R.sub.10' are each independently --X--Y-Z;
X is a bond, --C(H)(R.sub.6)--, --O--, --S--, or --N(R.sub.11)--;
R.sub.11 is: hydrogen-, (C1-C12)-aliphatic-, (C6-C10)-aryl-,
(C6-C10)-aryl-(C1-C12)aliphatic-, (C3-C10)-cycloalkyl- or
cycloalkenyl-, [(C3-C10)-cycloalkyl- or
cycloalkenyl]-(C1-C12)-aliphatic-, (C3-C10)-heterocyclyl-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, (C5-C10)-heteroaryl-, or
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-, wherein up to 3 aliphatic
carbon atoms in each R.sub.11 may be optionally replaced with S,
--S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)-- in a chemically
stable arrangement; wherein R.sub.11 may be optionally substituted
with up to 3 J substituents; or wherein R.sub.11 and Z together
with the atoms to which they are bound, optionally form a nitrogen
containing 5-7-membered mono- or 6-11-membered bicyclic ring system
optionally substituted with up to 3 J substitutents, wherein up to
3 ring atoms in said ring system may be optionally replaced with O,
NH, S, SO, or SO.sub.2 in a chemically stable arrangement; Y is a
bond, --CH.sub.2--, --C(O)--, --C(O)C(O)--, --S(O)--, S(O).sub.2--,
or --S(O)(NR.sub.12)--; R.sub.12 is: hydrogen-,
(C1-C12)-aliphatic-, (C6-C10)-aryl-,
(C6-C10)-aryl-(C1-C12)aliphatic-, (C3-C10)-cycloalkyl- or
cycloalkenyl-, [(C3-C10)-cycloalkyl- or
cycloalkenyl]-(C1-C12)-aliphatic-, (C3-C10)-heterocyclyl-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, (C5-C10)-heteroaryl-, or
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-, wherein up to 3 aliphatic
carbon atoms in each R.sub.12 may be optionally replaced with S,
--S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)--, in a
chemically stable arrangement; wherein R.sub.12 may be optionally
substituted with up to 3 J substituents; Z is: hydrogen-,
(C1-C12)-aliphatic-, (C3-C10)-cycloalkyl- or -cycloalkenyl-,
[(C3-C10)-cycloalkyl or -cycloalkenyl]-(C1-C12)-aliphatic-,
(C6-C10)-aryl-, (C6-C10)-aryl-(C1-C12)aliphatic-,
(C3-C10)-heterocyclyl-, (C3-C10)-heterocyclyl-(C1-C12)aliphatic-,
(C5-C10)-heteroaryl-, or (C5-C10)-heteroaryl-(C1-C12)-aliphatic-;
wherein up to three aliphatic carbon atoms in Z may be optionally
replaced with S, --S(O)--, --S(O).sub.2--, --O--, --N--, or
--N(H)--, in a chemically stable arrangement; wherein any ring may
be optionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,
(C3-C10)cycloalkyl, or (C3-C10)heterocyclyl; wherein Z may be
independently and optionally substituted with up to 3 substituents
independently selected from J; V is --C(O)--, --S(O)--, or
--S(O).sub.2--; R is --C(O)--, --S(O)--, --S(O).sub.2--,
--N(R.sub.12)--, --O--, or a bond; T is: (C1-C12)-aliphatic-;
(C6-C10)-aryl-, (C6-C10)-aryl-(C1-C12)aliphatic-,
(C3-C10)-cycloalkyl or -cycloalkenyl-, [(C3-C10)-cycloalkyl or
-cycloalkenyl]-(C1-C12)-aliphatic-, (C3-C10)-heterocyclyl-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, (C5-C10)-heteroaryl-, or
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; wherein up to 3 aliphatic
carbon atoms in T may be replaced with S, --S(O)--, --S(O).sub.2--,
--O--, --N--, or --N(H)--, in a chemically stable arrangement;
wherein each T may be optionally substituted with up to 3 J
substituents; or T is selected from --N(R.sub.6)(R.sub.6'); and
R.sub.6' is hydrogen-, (C1-C12)-aliphatic-, (C6-C10)-aryl-,
(C6-C10)-aryl-(C1-C12)aliphatic-, (C3-C10)-cycloalkyl- or
cycloalkenyl-, [(C3-C10)-cycloalkyl- or
cycloalkenyl]-(C1-C12)-aliphatic-, (C3-C10)-heterocyclyl-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, (C5-C10)-heteroaryl-, or
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-, or wherein up to 3
aliphatic carbon atoms in each R.sub.6' may be optionally replaced
with S, --S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)-- in a
chemically stable arrangement; wherein R.sub.6' may be optionally
substituted with up to 3 J substituents; or R.sub.6 and R.sub.6',
together with the nitrogen atom to which they are bound, may
optionally form a (C3-C10)-heterocyclic ring system wherein said
ring system may be optionally substituted with up to 3 substituents
independently selected from J.
2. The compound according to claim 1, wherein the ##STR00070##
radical is, ##STR00071## wherein; in R', R.sub.10, and R.sub.10', X
and Y are both a bond and Z is hydrogen; and in R.sub.9'; X is a
bond; Y is a bond, --CH.sub.2--, or --C(O)--; and Z is
(C1-C12)-aliphatic-, (C3-C10)-cycloalkyl- or -cycloalkenyl-,
[(C3-C10)-cycloalkyl or -cycloalkenyl]-(C1-C12)-aliphatic-,
(C6-C10)-aryl-, (C6-C10)-aryl-(C1-C12)aliphatic-,
(C3-C10)-heterocyclyl-, (C3-C10)-heterocyclyl-(C1-C12)aliphatic-,
(C5-C10)-heteroaryl-, or (C5-C10)-heteroaryl-(C1-C12)-aliphatic-;
wherein up to three aliphatic carbon atoms in Z may be optionally
replaced with S, --S(O)--, --S(O).sub.2--, --O--, --N--, or
--N(H)--, in a chemically stable arrangement; wherein any ring may
be optionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,
(C3-C10)cycloalkyl, or (C3-C10)heterocyclyl; wherein Z may be
independently and optionally substituted with up to 3 substituents
independently selected from J.
3. The compound according to claim 2, wherein in R.sub.9'; X is a
bond; Y is a bond; and Z is (C1-C12)-aliphatic-,
(C3-C10)-cycloalkyl- or -cycloalkenyl-, [(C3-C10)-cycloalkyl or
-cycloalkenyl]-(C1-C12)-aliphatic-, (C6-C10)-aryl-,
(C6-C10)-aryl-(C1-C12)aliphatic-, (C5-C10)-heteroaryl-, or
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; wherein up to three
aliphatic carbon atoms in Z may be optionally replaced with S,
--S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)--, in a
chemically stable arrangement; wherein any ring may be optionally
fused to a (C6-C10)aryl, (C5-C10)heteroaryl, (C3-C10)cycloalkyl, or
(C3-C10)heterocyclyl; wherein Z may be independently and optionally
substituted with up to 3 substituents independently selected from
J.
4. The compound according to claim 3, wherein in R.sub.9'; X is a
bond; Y is a bond; and Z is (C1-C12)-aliphatic-,
(C3-C10)-cycloalkyl- or -cycloalkenyl-, [(C3-C10)-cycloalkyl or
-cycloalkenyl]-(C1-C12)-aliphatic-, or
(C6-C10)-aryl-(C1-C12)aliphatic-, wherein up to three aliphatic
carbon atoms in Z may be optionally replaced with S, --S(O)--,
--S(O).sub.2--, --O--, --N--, or --N(H)--, in a chemically stable
arrangement; wherein Z may be independently and optionally
substituted with up to 3 substituents independently selected from
J.
5. The compound according to claim 4, wherein R.sub.9' is
##STR00072##
6. The compound according to claim 5, wherein R.sub.9' is
##STR00073##
7. The compound according to claim 6, wherein R.sub.9' is
ethyl.
8. The compound according to claim 1, wherein in R.sub.9, R.sub.10,
and R.sub.10', X and Y are both a bond and Z is hydrogen; and in
R.sub.9'; X is a bond; Y is --C(O)--; and Z is (C1-C12)-aliphatic-,
or (C3-C10)-heterocyclyl-(C1-C12)aliphatic-; wherein up to three
aliphatic carbon atoms in Z may be optionally replaced with S,
--S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)--, in a
chemically stable arrangement; wherein any ring may be optionally
fused to a (C6-C10)aryl, (C5-C10)heteroaryl, (C3-C10)cycloalkyl, or
(C3-C10)heterocyclyl; wherein Z may be independently and optionally
substituted with up to 3 substituents independently selected from
J.
9. The compound according to claim 8, wherein Z is
--O--(C1-C6)-aliphatic or --N(R').sub.2, wherein the two R' groups
bound to the nitrogen atom may optionally form a 3- to 7-membered
saturated or partially unsaturated ring system wherein up to 3 ring
atoms may be optionally replaced with a heteroatom independently
selected from N, NH, O, S, SO, and SO.sub.2, wherein said ring
system may be optionally fused to a (C6-C10)aryl,
(C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a (C3-C10)heterocyclyl,
wherein any ring has up to 3 substituents selected independently
from J.
10. The compound according to claim 8, wherein Z is --N(R').sub.2,
wherein the two R' groups bound to the nitrogen atom may optionally
form a 3- to 7-membered saturated or partially unsaturated ring
system wherein up to 3 ring atoms may be optionally replaced with a
heteroatom independently selected from N, NH, O, S, SO, and
SO.sub.2, wherein said ring system may be optionally fused to a
(C6-C10)aryl, (C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a
(C3-C10)heterocyclyl, wherein any ring has up to 3 substituents
selected independently from J.
11. The compound according to claim 1, wherein in R', and R.sub.10,
X and Y are a bond and Z is hydrogen; and in each of R.sub.9 and
R.sub.10' independently; X is a bond; Y is a bond; and Z is
(C1-C12)-aliphatic-, (C3-C10)-cycloalkyl- or -cycloalkenyl-,
[(C3-C10)-cycloalkyl or -cycloalkenyl]-(C1-C12)-aliphatic-,
(C6-C10)-aryl-, (C6-C10)-aryl-(C1-C12)aliphatic-,
(C3-C10)-heterocyclyl-, (C3-C10)-heterocyclyl-(C1-C12)aliphatic-,
(C5-C10)-heteroaryl-, or (C5-C10)-heteroaryl-(C1-C12)-aliphatic-;
wherein up to three aliphatic carbon atoms in Z may be optionally
replaced with S, --S(O)--, --S(O).sub.2--, --O--, --N--, or
--N(H)--, in a chemically stable arrangement; wherein any ring may
be optionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,
(C3-C10)cycloalkyl, or (C3-C10)heterocyclyl; wherein Z may be
independently and optionally substituted with up to 3 substituents
independently selected from J.
12. The compound according to claim 11, wherein Z, in each of
R.sub.9' and R.sub.10' independently, is (C1-C12)-aliphatic-,
(C3-C10)-cycloalkyl- or -cycloalkenyl-, or [(C3-C10)-cycloalkyl or
-cycloalkenyl]-(C1-C12)-aliphatic-; wherein up to three aliphatic
carbon atoms in Z may be optionally replaced with S, --S(O)--,
--S(O).sub.2--, --O--, --N--, or --N(H)--, in a chemically stable
arrangement; wherein Z may be independently and optionally
substituted with up to 3 substituents independently selected from
J.
13. The compound according to claim 12, wherein Z, in each of
R.sub.9' and R.sub.10' independently, is (C1-C6)-aliphatic-.
14. The compound according to claim 1, wherein in R.sub.10, and
R.sub.10', X and Y are a bond and Z is hydrogen; and in each of
R.sub.9 and R.sub.9'; X is a bond, Y is a bond, and Z is
(C1-C6)-aliphatic-, wherein Z may be independently and optionally
substituted with up to 3 substituents independently selected from
J.
15. The compound according to claim 1, wherein W is: ##STR00074##
wherein in the W, the NR.sub.6R.sub.6 is selected from --NH--(C1-C6
aliphatic), --NH-- (C3-C6 cycloalkyl), --NH--CH(CH.sub.3)-aryl, or
--NH--CH(CH.sub.3)-heteroaryl, wherein said aryl or said heteroaryl
is optionally substituted with up to 3 halogens.
16. The compound according to claim 15, wherein in the W, the
NR.sub.6R.sub.6 is: ##STR00075##
17. The compound according to claim 16, wherein in the W, the
NR.sub.6R.sub.6 is: ##STR00076##
18. The compound according to claim 17, wherein in the W, the
NR.sub.6R.sub.6 is: ##STR00077##
19. The compound according to claim 18, wherein in the W, the
NR.sub.6R.sub.6 is: ##STR00078##
20. The compound according to claim 1, wherein R.sub.5' is hydrogen
and R.sub.5 is: ##STR00079##
21. The compound according to claim 20, wherein R.sub.5' is
hydrogen and R.sub.5 is: ##STR00080##
22. The compound according to claim 1, wherein R.sub.2, R.sub.4,
and R.sub.7 are each independently H, methyl, ethyl, or propyl.
23. The compound according to claim 22, wherein R.sub.2, R.sub.4,
and R.sub.7 are each hydrogen.
24. The compound according to claim 1, wherein R.sub.3 is:
##STR00081##
25. The compound according to claim 24, wherein R.sub.3 is:
##STR00082##
26. The compound according to claim 25, wherein R.sub.3 is:
##STR00083##
27. The compound according to claim 1, wherein R.sub.1 is:
##STR00084##
28. The compound according to claim 27, wherein R.sub.1 is:
##STR00085##
29. The compound according to claim 18, wherein R.sub.1 is
isopropyl or cyclohexyl.
30. The compound according to claim 1, wherein the ##STR00086##
radical is: ##STR00087## wherein: R.sub.6, R.sub.6', R.sub.7, and
R.sub.12, are as defined in claim 1.
31. The compound according to claim 30, wherein in the ##STR00088##
radical; R.sub.6' and R.sub.7 are both hydrogen; R.sub.6 is:
(C1-C12)-aliphatic-; (C6-C10)-aryl-,
(C6-C10)-aryl-(C1-C12)aliphatic-, (C3-C10)-cycloalkyl or
-cycloalkenyl-, [(C3-C10)-cycloalkyl or
-cycloalkenyl]-(C1-C12)-aliphatic-, (C3-C10)-heterocyclyl-,
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, (C5-C10)-heteroaryl-, or
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; wherein up to 3 aliphatic
carbon atoms in R.sub.6 may be optionally replaced by S, --S(O)--,
--S(O).sub.2--, --O--, --N--, or --N(H)--, in a chemically stable
arrangement; and wherein R.sub.6 may be optionally substituted with
up to 3 substituents independently selected from J; and R.sub.12 is
as defined in claim 1.
32. The compound according to claim 31, wherein; R.sub.6 is:
(C1-C12)-aliphatic-; (C6-C10)-aryl-(C1-C12)aliphatic-, or
(C3-C10)-cycloalkyl or -cycloalkenyl-; wherein up to 3 aliphatic
carbon atoms in R.sub.6 may be optionally replaced by S, --S(O)--,
--S(O).sub.2--, --O--, --N--, or --N(H)--, in a chemically stable
arrangement; wherein R.sub.6 may be optionally substituted with up
to 3 substituents independently selected from J; and R.sub.12 is as
defined in claim 1.
33. The compound according to claim 32, wherein the radical is:
##STR00089##
34. The compound according to claim 33, wherein the ##STR00090##
radical is: ##STR00091##
35. The compound according to claim 1, wherein; V is --C(O)--; and
R is a bond.
36. The compound according to claim 1, wherein; V is --C(O)--; R is
a bond; and T is: (C3-C10)-heterocyclyl- or (C5-C10)heteroaryl-;
wherein each T is optionally substituted with up to 3 J
substituents.
37. The compound according to claim 36, wherein T is
(C5-C6)heterocyclyl- or (C5-C6)heteroaryl-; wherein each T is
optionally substituted with up to 3 J substituents.
38. The compound according to claim 37, wherein T is: ##STR00092##
wherein: Z' is independently O, S, NR', or C(R').sub.2.
39. The compound according to claim 38, wherein T is:
##STR00093##
40. The compound according to claim 1, wherein the compound is:
##STR00094## ##STR00095## ##STR00096## ##STR00097## ##STR00098##
##STR00099## ##STR00100## ##STR00101## ##STR00102## ##STR00103##
##STR00104## ##STR00105## ##STR00106## ##STR00107## ##STR00108##
##STR00109## ##STR00110## ##STR00111## ##STR00112##
41. A pharmaceutical composition comprising a compound according to
claim 1 or a pharmaceutically acceptable salt or mixtures thereof
in an amount effective to inhibit a serine protease; and a
acceptable carrier, adjuvant or vehicle.
42.-45. (canceled)
46. A method of inhibiting the activity of a serine protease
comprising the step of contacting said serine protease with a
compound according to claim 1.
47. (canceled)
48. A method of treating an HCV infection in a patient comprising
the step of administering to said patient a composition according
to claim 42.
49.-50. (canceled)
51. A method of eliminating or reducing HCV contamination of a
biological sample or medical or laboratory equipment, comprising
the step of contacting said biological sample or medical or
laboratory equipment with a composition according to claim 41.
52.-53. (canceled)
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] The present application claims the benefit of U.S.
Provisional Application No. 60/513,765, filed Oct. 23, 2003,
entitled "Inhibitors of Serine Proteases, Particularly HCV NS3-NS4A
Protease", the entire contents of which is hereby incorporated by
reference. The present application also claims the benefit of U.S.
patent application Ser. No. 10/412,600, filed Apr. 11, 2003,
entitled "Inhibitors of Serine Proteases, Particularly HCV NS3-NS4A
Protease", the entire contents of which is hereby incorporated by
reference.
TECHNICAL FIELD OF THE INVENTION
[0002] The present invention relates to compounds that inhibit
serine protease activity, particularly the activity of hepatitis C
virus NS3-NS4A protease. As such, they act by interfering with the
life cycle of the hepatitis C virus and are also useful as
antiviral agents. The invention further relates to pharmaceutical
compositions comprising these compounds either for ex vivo use or
for administration to a patient suffering from HCV infection. The
invention also relates to processes for preparing the compounds and
methods of treating an HCV infection in a patient by administering
a pharmaceutical composition comprising a compound of this
invention.
BACKGROUND OF THE INVENTION
[0003] Infection by hepatitis C virus ("HCV") is a compelling human
medical problem. HCV is recognized as the causative agent for most
cases of non-A, non-B hepatitis, with an estimated human
sero-prevalence of 3% globally [A. Alberti et al., "Natural History
of Hepatitis C," J. Hepatology, 31., (Suppl. 1), pp. 17-24 (1999)].
Nearly four million individuals may be infected in the United
States alone [M. J. Alter et al., "The Epidemiology of Viral
Hepatitis in the United States, Gastroenterol. Clin. North Am., 23,
pp. 437-455 (1994); M. J. Alter "Hepatitis C Virus Infection in the
United States," J. Hepatology, 31., (Suppl. 1), pp. 88-91
(1999)].
[0004] Upon first exposure to HCV only about 20% of infected
individuals develop acute clinical hepatitis while others appear to
resolve the infection spontaneously. In almost 70% of instances,
however, the virus establishes a chronic infection that persists
for decades [S. Iwarson, "The Natural Course of Chronic Hepatitis,"
FEMS Microbiology Reviews, 14, pp. 201-204 (1994); D. Lavanchy,
"Global Surveillance and Control of Hepatitis C," J. Viral
Hepatitis, 6, pp. 35-47 (1999)]. This usually results in recurrent
and progressively worsening liver inflammation, which often leads
to more severe disease states such as cirrhosis and hepatocellular
carcinoma [M. C. Kew, "Hepatitis C and Hepatocellular Carcinoma",
FEMS Microbiology Reviews, 14, pp. 211-220 (1994); I. Saito et.
al., "Hepatitis C Virus Infection is Associated with the
Development of Hepatocellular Carcinoma," Proc. Natl. Acad. Sci.
USA, 87, pp. 6547-6549 (1990)]. Unfortunately, there are no broadly
effective treatments for the debilitating progression of chronic
HCV.
[0005] The HCV genome encodes a polyprotein of 3010-3033 amino
acids [Q. L. Choo, et. al., "Genetic Organization and Diversity of
the Hepatitis C Virus." Proc. Natl. Acad. Sci. USA, 88, pp.
2451-2455 (1991); N. Kato et al., "Molecular Cloning of the Human
Hepatitis C Virus Genome From Japanese Patients with Non-A, Non-B
Hepatitis," Proc. Natl. Acad. Sci. USA, 87, pp. 9524-9528 (1990);
A. Takamizawa et. al., "Structure and Organization of the Hepatitis
C Virus Genome Isolated From Human Carriers," J. Virol., 65, pp.
1105-1113 (1991)]. The HCV nonstructural (NS) proteins are presumed
to provide the essential catalytic machinery for viral replication.
The NS proteins are derived by proteolytic cleavage of the
polyprotein [R. Bartenschlager et. al., "Nonstructural Protein 3 of
the Hepatitis C Virus Encodes a Serine-Type Proteinase Required for
Cleavage at the NS3/4 and NS4/5 Junctions," J. Virol., 67, pp.
3835-3844 (1993); A. Grakoui et. al., "Characterization of the
Hepatitis C Virus-Encoded Serine Proteinase: Determination of
Proteinase-Dependent Polyprotein Cleavage Sites," J. Virol., 67,
pp. 2832-2843 (1993); A. Grakoui et. al., "Expression and
Identification of Hepatitis C Virus Polyprotein Cleavage Products,"
J. Virol., 67, pp. 1385-1395 (1993); L. Tomei et. al., "NS3 is a
serine protease required for processing of hepatitis C virus
polyprotein", J. Virol., 67, pp. 4017-4026 (1993)].
[0006] The HCV NS protein 3 (NS3) contains a serine protease
activity that helps process the majority of the viral enzymes, and
is thus considered essential for viral replication and infectivity.
It is known that mutations in the yellow fever virus NS3 protease
decrease viral infectivity [Chambers, T. J. et. al., "Evidence that
the N-terminal Domain of Nonstructural Protein NS3 From Yellow
Fever Virus is a Serine Protease Responsible for Site-Specific
Cleavages in the Viral Polyprotein", Proc. Natl. Acad. Sci. USA,
87, pp. 8898-8902 (1990)]. The first 181 amino acids of NS3
(residues 1027-1207 of the viral polyprotein) have been shown to
contain the serine protease domain of NS3 that processes all four
downstream sites of the HCV polyprotein [C. Lin et al., "Hepatitis
C Virus NS3 Serine Proteinase: Trans-Cleavage Requirements and
Processing Kinetics", J. Virol., 68, pp. 8147-8157 (1994)].
[0007] The HCV NS3 serine protease and its associated cofactor,
NS4A, helps process all of the viral enzymes, and is thus
considered essential for viral replication. This processing appears
to be analogous to that carried out by the human immunodeficiency
virus aspartyl protease, which is also involved in viral enzyme
processing. HIV protease inhibitors, which inhibit viral protein
processing, are potent antiviral agents in man, indicating that
interrupting this stage of the viral life cycle results in
therapeutically active agents. Consequently HCV NS3 serine protease
is also an attractive target for drug discovery.
[0008] Furthermore, the current understanding of HCV has not led to
any other satisfactory anti-HCV agents or treatments. Until
recently, the only established therapy for HCV disease was
interferon treatment. However, interferons have significant side
effects [M. A. Wlaker et al., "Hepatitis C Virus: An Overview of
Current Approaches and Progress," DDT, 4, pp. 518-29 (1999); D.
Moradpour et al., "Current and Evolving Therapies for Hepatitis C,"
Eur. J. Gastroenterol. Hepatol., 11, pp. 1199-1202 (1999); H. L. A.
Janssen et al. "Suicide Associated with Alfa-Interferon Therapy for
Chronic Viral Hepatitis," J. Hepatol., 21, pp. 241-243 (1994); P.
F. Renault et al., "Side Effects of Alpha Interferon," Seminars in
Liver Disease, 9, pp. 273-277. (1989)] and induce long term
remission in only a fraction (.about.25%) of cases [O. Weiland,
"Interferon Therapy in Chronic Hepatitis C Virus Infection", FEMS
Microbiol. Rev., 14, pp. 279-288 (1994)]. Recent introductions of
the pegylated forms of interferon (PEG-Intron.RTM. and
Pegasys.RTM.) and the combination therapy of ribavirin and
pegylated interferon (Rebetrol.RTM.) have resulted in only modest
improvements in remission rates and only partial reductions in side
effects. Moreover, the prospects for effective anti-HCV vaccines
remain uncertain.
[0009] Thus, there is a need for more effective anti-HCV therapies.
Such inhibitors would have therapeutic potential as protease
inhibitors, particularly as serine protease inhibitors, and more
particularly as HCV NS3 protease inhibitors. Specifically, such
compounds may be useful as antiviral agents, particularly as
anti-HCV agents.
SUMMARY OF THE INVENTION
[0010] The present invention provides a compound of formula I:
##STR00002##
or a pharmaceutically acceptable salt, or mixtures thereof,
wherein: [0011] W is:
[0011] ##STR00003## [0012] wherein each R.sub.6 is independently:
[0013] hydrogen-, [0014] (C1-C12)-aliphatic-, [0015]
(C6-C10)-aryl-, [0016] (C6-C10)-aryl-(C1-C12)aliphatic-, [0017]
(C3-C10)-cycloalkyl- or cycloalkenyl-, [0018] [(C3-C10)-cycloalkyl-
or cycloalkenyl]-(C1-C12)-aliphatic-, [0019]
(C3-C10)-heterocyclyl-, [0020]
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, [0021]
(C5-C10)-heteroaryl-, or [0022]
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-, or [0023] wherein up to 3
aliphatic carbon atoms in each R.sub.6 may be optionally replaced
with S, --S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)-- in a
chemically stable arrangement; [0024] wherein R.sub.6 may be
optionally substituted with up to 3 J substituents; or [0025] two
R.sub.6 groups, together with the nitrogen atom to which they are
bound, may optionally form a 5- to 6-membered aromatic or a 3- to
7-membered saturated or partially unsaturated ring system wherein
up to 3 ring atoms may be optionally replaced with N, NH, O, S, SO,
and SO.sub.2, wherein said ring system may be optionally fused to a
(C6-C10)aryl, (C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a
(C3-C10)heterocyclyl, wherein any ring has up to 3 substituents
selected independently from J; [0026] wherein each R.sub.8 is
independently --OR'; or the R.sub.8 groups together with the boron
atom, may optionally form a (C3-C10)-membered heterocyclic ring
having, in addition to the boron, up to 3 ring atoms optionally
replaced with N, NH, O, S, SO, and SO.sub.2; [0027] J is halogen,
--OR', --NO.sub.2, --CN, --CF.sub.3, --OCF.sub.3, --R', oxo,
thioxo, .dbd.N(R'), .dbd.N(OR'), 1,2-methylenedioxy,
1,2-ethylenedioxy, --N(R').sub.2, --SR', --SOR', --SO.sub.2R',
--SO.sub.2N(R').sub.2, --SO.sub.3R', --C(O)R', --C(O)C(O)R',
--C(O)C(O)OR', --C(O)C(O)NR', --C(O)CH.sub.2C(O)R', --C(S)R',
--C(S)OR', --C(O)OR', --OC(O)R', --C(O)N(R').sub.2,
--OC(O)N(R').sub.2, --C(S)N(R').sub.2,
--(CH.sub.2).sub.0-2NHC(O)R', --N(R')N(R')COR',
--N(R')N(R')C(O)OR', --N(R')N(R')CON(R').sub.2, --N(R')SO.sub.2R',
--N(R')SO.sub.2N(R').sub.2, --N(R')C(O)OR', --N(R')C(O)R',
--N(R')C(S)R', --N(R')C(O)N(R').sub.2, --N(R')C(S)N(R').sub.2,
--N(COR')COR', --N(OR')R', --C(.dbd.NH)N(R').sub.2, --C(O)N(OR')R',
--C(.dbd.NOR')R', --OP(O)(OR').sub.2, --P(O)(R').sub.2,
--P(O)(OR').sub.2, or --P(O)(H)(OR'); wherein; [0028] R' is
independently selected from: [0029] hydrogen-, [0030]
(C1-C12)-aliphatic-, [0031] (C3-C10)-cycloalkyl- or -cycloalkenyl-,
[0032] [(C3-C10)-cycloalkyl or -cycloalkenyl]-(C1-C12)-aliphatic-,
[0033] (C6-C10)-aryl-, [0034] (C6-C10)-aryl-(C1-C12)aliphatic-,
[0035] (C3-C10)-heterocyclyl-, [0036]
(C3-C10)-heterocyclyl-(C1-C12)aliphatic-, [0037]
(C5-C10)-heteroaryl-, and [0038]
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; [0039] wherein up to 5
atoms in R' may be optionally and independently substituted with J;
[0040] wherein two R' groups bound to the same atom may optionally
form a 5- to 6-membered aromatic or a 3- to 7-membered saturated or
partially unsaturated ring system wherein up to 3 ring atoms may be
optionally replaced with a heteroatom independently selected from
N, NH, O, S, SO, and SO.sub.2, wherein said ring system may be
optionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,
(C3-C10)cycloalkyl, or a (C3-C10)heterocyclyl, wherein any ring has
up to 3 substituents selected independently from J; [0041] R.sub.5
and R.sub.5' are each independently hydrogen or (C1-C12)-aliphatic,
wherein any hydrogen may be optionally replaced with halogen;
wherein any terminal carbon atom of R.sub.5 may be optionally
substituted with sulfhydryl or hydroxy; or R.sub.5 is Ph or
--CH.sub.2Ph and R.sub.5' is H, wherein said Ph or --CH.sub.2Ph
group may be optionally substituted with up to 3 substituents
independently selected from J; or [0042] R.sub.5 and R.sub.5'
together with the atom to which they are bound may optionally form
a 3- to 6-membered saturated or partially unsaturated ring system
wherein up to 2 ring atoms may be optionally replaced with N, NH,
O, SO, or SO.sub.2; wherein said ring system has up to 2
substituents selected independently from J; [0043] R.sub.2,
R.sub.4, and R.sub.7 are each independently: [0044] hydrogen-,
[0045] (C1-C12)-aliphatic-, [0046]
(C3-C10)-cycloalkyl-(C1-C12)-aliphatic-, or [0047]
(C6-C10)-aryl-(C1-C12)-aliphatic-; [0048] wherein up to two
aliphatic carbon atoms in each of R.sub.2, R.sub.4, and R.sub.7 may
be optionally replaced with S, --S(O)--, --S(O).sub.2--, --O--,
--N--, or --N(H)-- in a chemically stable arrangement; [0049]
wherein each of R.sub.2, R.sub.4, and R.sub.7 may be independently
and optionally substituted with up to 3 substituents independently
selected from J; [0050] R.sub.1 and R.sub.3 are each independently:
[0051] (C1-C12)-aliphatic-, [0052] (C3-C10)-cycloalkyl- or
-cycloalkenyl-, [0053] [(C3-C10)-cycloalkyl- or
-cycloalkenyl]-(C1-C12)-aliphatic-, [0054]
(C6-C10)-aryl-(C1-C12)aliphatic-, or [0055]
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; [0056] wherein up to 3
aliphatic carbon atoms in each of R.sub.1 and R.sub.3 may be
optionally replaced with S, --S(O)--, --S(O).sub.2--, --O--, --N--,
or --N(H)-- in a chemically stable arrangement; [0057] wherein each
of R.sub.1 and R.sub.3 may be independently and optionally
substituted with up to 3 substituents independently selected from
J; [0058] R.sub.9, R.sub.9', R.sub.10, and R.sub.10' are each
independently --X--Y-Z; [0059] X is a bond, --C(H)(R.sub.6)--,
--O--, --S--, or --N(R.sub.11)--; [0060] R.sub.11 is: [0061]
hydrogen-, [0062] (C1-C12)-aliphatic-, [0063] (C6-C10)-aryl-,
[0064] (C6-C10)-aryl-(C1-C12)aliphatic-, [0065]
(C3-C10)-cycloalkyl- or cycloalkenyl-, [0066] [(C3-C10)-cycloalkyl-
or cycloalkenyl]-(C1-C12)-aliphatic-, [0067]
(C3-C10)-heterocyclyl-, [0068]
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, [0069]
(C5-C10)-heteroaryl-, or [0070]
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-, [0071] wherein up to 3
aliphatic carbon atoms in each R.sub.11 may be optionally replaced
with S, --S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)-- in a
chemically stable arrangement; [0072] wherein R.sub.11 may be
optionally substituted with up to 3 J substituents; or [0073]
wherein R.sub.11 and Z together with the atoms to which they are
bound, optionally form a nitrogen containing 5-7-membered mono- or
6-11-membered bicyclic ring system optionally substituted with up
to 3 J substitutents, wherein up to 3 ring atoms in said ring
system may be optionally replaced with O, NH, S, SO, or SO.sub.2 in
a chemically stable arrangement; [0074] Y is a bond, --CH.sub.2--,
--C(O)--, --C(O)C(O)--, --S(O)--, S(O).sub.2--, or
--S(O)(NR.sub.12)--; [0075] R.sub.12 is: [0076] hydrogen-, [0077]
(C1-C12)-aliphatic-, [0078] (C6-C10)-aryl-, [0079]
(C6-C10)-aryl-(C1-C12)aliphatic-, [0080] (C3-C10)-cycloalkyl- or
cycloalkenyl-, [0081] [(C3-C10)-cycloalkyl- or
cycloalkenyl]-(C1-C12)-aliphatic-, [0082] (C3-C10)-heterocyclyl-,
[0083] (C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, [0084]
(C5-C10)-heteroaryl-, or [0085]
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-, [0086] wherein up to 3
aliphatic carbon atoms in each R.sub.12 may be optionally replaced
with S, --S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)--, in a
chemically stable arrangement; [0087] wherein R.sub.12 may be
optionally substituted with up to 3 J substituents; [0088] Z is:
[0089] hydrogen-, [0090] (C1-C12)-aliphatic-, [0091]
(C3-C10)-cycloalkyl- or -cycloalkenyl-, [0092] [(C3-C10)-cycloalkyl
or -cycloalkenyl]-(C1-C12)-aliphatic-, [0093] (C6-C10)-aryl-,
[0094] (C6-C10)-aryl-(C1-C12)aliphatic-, [0095]
(C3-C10)-heterocyclyl-, [0096]
(C3-C10)-heterocyclyl-(C1-C12)aliphatic-, [0097]
(C5-C10)-heteroaryl-, or [0098]
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; [0099] wherein up to three
aliphatic carbon atoms in Z may be optionally replaced with S,
--S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)--, in a
chemically stable arrangement; [0100] wherein any ring may be
optionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,
(C3-C10)cycloalkyl, or (C3-C10)heterocyclyl; [0101] wherein Z may
be independently and optionally substituted with up to 3
substituents independently selected from J; [0102] V is --C(O)--,
--S(O)--, or --S(O).sub.2--; [0103] R is --C(O)--, --S(O)--,
--S(O).sub.2--, --N(R.sub.12)--, --O--, or a bond; [0104] T is:
[0105] (C1-C12)-aliphatic-; [0106] (C6-C10)-aryl-, [0107]
(C6-C10)-aryl-(C1-C12)aliphatic-, [0108] (C3-C10)-cycloalkyl or
-cycloalkenyl-, [0109] [(C3-C10)-cycloalkyl or
-cycloalkenyl]-(C1-C12)-aliphatic-, [0110] (C3-C10)-heterocyclyl-,
[0111] (C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, [0112]
(C5-C10)-heteroaryl-, or [0113]
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; [0114] wherein up to 3
aliphatic carbon atoms in T may be replaced with S, --S(O)--,
--S(O).sub.2--, --O--, --N--, or --N(H)--, in a chemically stable
arrangement; [0115] wherein each T may be optionally substituted
with up to 3 J substituents; or [0116] T is selected from
--N(R.sub.6)(R.sub.6'); and [0117] R.sub.6' is [0118] hydrogen-,
[0119] (C1-C12)-aliphatic-, [0120] (C6-C10)-aryl-, [0121]
(C6-C10)-aryl-(C1-C12)aliphatic-, [0122] (C3-C10)-cycloalkyl- or
cycloalkenyl-, [0123] [(C3-C10)-cycloalkyl- or
cycloalkenyl]-(C1-C12)-aliphatic-, [0124] (C3-C10)-heterocyclyl-,
[0125] (C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, [0126]
(C5-C10)-heteroaryl-, or [0127]
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-, or [0128] wherein up to 3
aliphatic carbon atoms in each R.sub.6' may be optionally replaced
with S, --S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)-- in a
chemically stable arrangement; [0129] wherein R.sub.6' may be
optionally substituted with up to 3 J substituents; or [0130]
R.sub.6 and R.sub.6', together with the nitrogen atom to which they
are bound, may optionally form a (C3-C10)-heterocyclic ring system
wherein said ring system may be optionally substituted with up to 3
substituents independently selected from J.
[0131] The invention also relates to processes for preparing the
above compounds and to compositions that comprise the above
compounds and the use thereof. Such compositions may be used to
pre-treat invasive devices to be inserted into a patient, to treat
biological samples, such as blood, prior to administration to a
patient, and for direct administration to a patient. In each case
the composition will be used to inhibit HCV replication and to
lessen the risk of or the severity of HCV infection.
DETAILED DESCRIPTION OF THE INVENTION
[0132] The present invention provides a compound of formula I:
##STR00004##
or a pharmaceutically acceptable salt, or mixtures thereof,
wherein: [0133] W is:
[0133] ##STR00005## [0134] wherein each R.sub.6 is independently:
[0135] hydrogen-, [0136] (C1-C12)-aliphatic-, [0137]
(C6-C10)-aryl-, [0138] (C6-C10)-aryl-(C1-C12)aliphatic-, [0139]
(C3-C10)-cycloalkyl- or cycloalkenyl-, [0140] [(C3-C10)-cycloalkyl-
or cycloalkenyl]-(C1-C12)-aliphatic-, [0141]
(C3-C10)-heterocyclyl-, [0142]
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, [0143]
(C5-C10)-heteroaryl-, or [0144]
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-, or [0145] wherein up to 3
aliphatic carbon atoms in each R.sub.6 may be optionally replaced
with S, --S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)-- in a
chemically stable arrangement; [0146] wherein R.sub.6 may be
optionally substituted with up to 3 J substituents; or [0147] two
R.sub.6 groups, together with the nitrogen atom to which they are
bound, may optionally form a 5- to 6-membered aromatic or a 3- to
7-membered saturated or partially unsaturated ring system wherein
up to 3 ring atoms may be optionally replaced with N, NH, O, S, SO,
and SO.sub.2, wherein said ring system may be optionally fused to a
(C6-C10)aryl, (C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a
(C3-C10)heterocyclyl, wherein any ring has up to 3 substituents
selected independently from J; [0148] wherein each R.sub.8 is
independently --OR'; or the R.sub.8 groups together with the boron
atom, may optionally form a (C3-C10)-membered heterocyclic ring
having, in addition to the boron, up to 3 ring atoms optionally
replaced with N, NH, O, S, SO, and SO.sub.2; [0149] J is halogen,
--OR', --NO.sub.2, --CN, --CF.sub.3, --OCF.sub.3, --R', oxo,
thioxo, .dbd.N(R'), .dbd.N(OR'), 1,2-methylenedioxy,
1,2-ethylenedioxy, --N(R').sub.2, --SR', --SOR', --SO.sub.2R',
--SO.sub.2N(R').sub.2, --SO.sub.3R', --C(O)R', --C(O)C(O)R',
--C(O)C(O)OR', --C(O)C(O)NR', --C(O)CH.sub.2C(O)R', --C(S)R',
--C(S)OR', --C(O)OR', --OC(O)R', --C(O)N(R').sub.2,
--OC(O)N(R').sub.2, --C(S)N(R').sub.2,
--(CH.sub.2).sub.0-2NHC(O)R', --N(R')N(R')COR',
--N(R')N(R')C(O)OR', --N(R')N(R')CON(R').sub.2, --N(R') SO.sub.2R',
--N(R')SO.sub.2N(R').sub.2, --N(R')C(O)OR', --N(R')C(O)R',
--N(R')C(S)R', --N(R')C(O)N(R').sub.2, --N(R')C(S)N(R').sub.2,
--N(COR')COR', --N(OR')R', --C(.dbd.NH)N(R').sub.2, --C(O)N(OR')R',
--C(.dbd.NOR')R', --OP(O)(OR').sub.2, --P(O)(R').sub.2,
--P(O)(OR').sub.2, or --P(O)(H)(OR'); wherein; [0150] R' is
independently selected from: [0151] hydrogen-, [0152]
(C1-C12)-aliphatic-, [0153] (C3-C10)-cycloalkyl- or -cycloalkenyl-,
[0154] [(C3-C10)-cycloalkyl or -cycloalkenyl]-(C1-C12)-aliphatic-,
[0155] (C6-C10)-aryl-, [0156] (C6-C10)-aryl-(C1-C12)aliphatic-,
[0157] (C3-C10)-heterocyclyl-, [0158]
(C3-C10)-heterocyclyl-(C1-C12)aliphatic-, [0159]
(C5-C10)-heteroaryl-, and (C5-C10)-heteroaryl-(C1-C12)-aliphatic-;
[0160] wherein up to 5 atoms in R' may be optionally and
independently substituted with J; [0161] wherein two R' groups
bound to the same atom may optionally form a 5- to 6-membered
aromatic or a 3- to 7-membered saturated or partially unsaturated
ring system wherein up to 3 ring atoms may be optionally replaced
with a heteroatom independently selected from N, NH, O, S, SO, and
SO.sub.2, wherein said ring system may be optionally fused to a
(C6-C10)aryl, (C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a
(C3-C10)heterocyclyl, wherein any ring has up to 3 substituents
selected independently from J; [0162] R.sub.5 and R.sub.5' are each
independently hydrogen or (C1-C12)-aliphatic, wherein any hydrogen
may be optionally replaced with halogen; wherein any terminal
carbon atom of R.sub.5 may be optionally substituted with
sulfhydryl or hydroxy; or R.sub.5 is Ph or --CH.sub.2Ph and R.sub.5
is H, wherein said Ph or --CH.sub.2Ph group may be optionally
substituted with up to 3 substituents independently selected from
J; or [0163] R.sub.5 and R.sub.5' together with the atom to which
they are bound may optionally form a 3- to 6-membered saturated or
partially unsaturated ring system wherein up to 2 ring atoms may be
optionally replaced with N, NH, O, SO, or SO.sub.2; wherein said
ring system has up to 2 substituents selected independently from J;
[0164] R.sub.2, R.sub.4, and R.sub.7 are each independently: [0165]
hydrogen-, [0166] (C1-C12)-aliphatic-, [0167]
(C3-C10)-cycloalkyl-(C1-C12)-aliphatic-, or [0168]
(C6-C10)-aryl-(C1-C12)-aliphatic-; [0169] wherein up to two
aliphatic carbon atoms in each of R.sub.2, R.sub.4, and R.sub.7 may
be optionally replaced with S, --S(O)--, --S(O).sub.2--, --O--,
--N--, or --N(H)-- in a chemically stable arrangement; [0170]
wherein each of R.sub.2, R.sub.4, and R.sub.7 may be independently
and optionally substituted with up to 3 substituents independently
selected from J; [0171] R.sub.1 and R.sub.3 are each independently:
[0172] (C1-C12)-aliphatic-, [0173] (C3-C10)-cycloalkyl- or
-cycloalkenyl-, [0174] [(C3-C10)-cycloalkyl- or
-cycloalkenyl]-(C1-C12)-aliphatic-, [0175]
(C6-C10)-aryl-(C1-C12)aliphatic-, or [0176]
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; [0177] wherein up to 3
aliphatic carbon atoms in each of R.sub.1 and R.sub.3 may be
optionally replaced with S, --S(O)--, --S(O).sub.2--, --O--, --N--,
or --N(H)-- in a chemically stable arrangement; [0178] wherein each
of R.sub.1 and R.sub.3 may be independently and optionally
substituted with up to 3 substituents independently selected from
J; [0179] R.sub.9, R.sub.9', R.sub.10, and R.sub.10' are each
independently --X--Y-Z; [0180] X is a bond, --C(H)(R.sub.6)--,
--O--, --S--, or --N(R.sub.11)--; [0181] R.sub.11 is: [0182]
hydrogen-, [0183] (C1-C12)-aliphatic-, [0184] (C6-C10)-aryl-,
[0185] (C6-C10)-aryl-(C1-C12)aliphatic-, [0186]
(C3-C10)-cycloalkyl- or cycloalkenyl-, [0187] [(C3-C10)-cycloalkyl-
or cycloalkenyl]-(C1-C12)-aliphatic-, [0188]
(C3-C10)-heterocyclyl-, [0189]
(C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, [0190]
(C5-C10)-heteroaryl-, or [0191]
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-, [0192] wherein up to 3
aliphatic carbon atoms in each R.sub.11 may be optionally replaced
with S, --S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)-- in a
chemically stable arrangement; [0193] wherein R.sub.11 may be
optionally substituted with up to 3 J substituents; or [0194]
wherein R.sub.11 and Z together with the atoms to which they are
bound, optionally form a nitrogen containing 5-7-membered mono- or
6-11-membered bicyclic ring system optionally substituted with up
to 3 J substitutents, wherein up to 3 ring atoms in said ring
system may be optionally replaced with O, NH, S, SO, or SO.sub.2 in
a chemically stable arrangement; [0195] Y is a bond, --CH.sub.2--,
--C(O)--, --C(O)C(O)--, --S(O)--, S(O).sub.2--, or
--S(O)(NR.sub.12)--; [0196] R.sub.12 is: [0197] hydrogen-, [0198]
(C1-C12)-aliphatic-, [0199] (C6-C10)-aryl-, [0200]
(C6-C10)-aryl-(C1-C12)aliphatic-, [0201] (C3-C10)-cycloalkyl- or
cycloalkenyl-, [0202] [(C3-C10)-cycloalkyl- or
cycloalkenyl]-(C1-C12)-aliphatic-, [0203] (C3-C10)-heterocyclyl-,
[0204] (C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, [0205]
(C5-C10)-heteroaryl-, or [0206]
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-, [0207] wherein up to 3
aliphatic carbon atoms in each R.sub.12 may be optionally replaced
with S, --S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)--, in a
chemically stable arrangement; [0208] wherein R.sub.12 may be
optionally substituted with up to 3 J substituents; [0209] Z is:
[0210] hydrogen-, [0211] (C1-C12)-aliphatic-, [0212]
(C3-C10)-cycloalkyl- or -cycloalkenyl-, [0213] [(C3-C10)-cycloalkyl
or -cycloalkenyl]-(C1-C12)-aliphatic-, [0214] (C6-C10)-aryl-,
[0215] (C6-C10)-aryl-(C1-C12)aliphatic-, [0216]
(C3-C10)-heterocyclyl-, [0217]
(C3-C10)-heterocyclyl-(C1-C12)aliphatic-, [0218]
(C5-C10)-heteroaryl-, or [0219]
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; [0220] wherein up to three
aliphatic carbon atoms in Z may be optionally replaced with S,
--S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)--, in a
chemically stable arrangement; [0221] wherein any ring may be
optionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,
(C3-C10)cycloalkyl, or (C3-C10)heterocyclyl; [0222] wherein Z may
be independently and optionally substituted with up to 3
substituents independently selected from J; [0223] V is --C(O)--,
--S(O)--, or --S(O).sub.2--; [0224] R is --C(O)--, --S(O)--,
--S(O).sub.2--, --N(R.sub.12)--, --O--, or a bond; [0225] T is:
[0226] (C1-C12)-aliphatic-; [0227] (C6-C10)-aryl-, [0228]
(C6-C10)-aryl-(C1-C12)aliphatic-, [0229] (C3-C10)-cycloalkyl or
-cycloalkenyl-, [0230] [(C3-C10)-cycloalkyl or
-cycloalkenyl]-(C1-C12)-aliphatic-, [0231] (C3-C10)-heterocyclyl-,
[0232] (C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, [0233]
(C5-C10)-heteroaryl-, or [0234]
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; [0235] wherein up to 3
aliphatic carbon atoms in T may be replaced with S, --S(O)--,
--S(O).sub.2--, --O--, --N--, or --N(H)--, in a chemically stable
arrangement; [0236] wherein each T may be optionally substituted
with up to 3 J substituents; or [0237] T is selected from
--N(R.sub.6)(R.sub.6'); and [0238] R.sub.6' is [0239] hydrogen-,
[0240] (C1-C12)-aliphatic-, [0241] (C6-C10)-aryl-, [0242]
(C6-C10)-aryl-(C1-C12)aliphatic-, [0243] (C3-C10)-cycloalkyl- or
cycloalkenyl-, [0244] [(C3-C10)-cycloalkyl- or
cycloalkenyl]-(C1-C12)-aliphatic-, [0245] (C3-C10)-heterocyclyl-,
[0246] (C3-C10)-heterocyclyl-(C1-C12)-aliphatic-, [0247]
(C5-C10)-heteroaryl-, or [0248]
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-, or [0249] wherein up to 3
aliphatic carbon atoms in each R.sub.6' may be optionally replaced
with S, --S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)-- in a
chemically stable arrangement; [0250] wherein R.sub.6 may be
optionally substituted with up to 3 J substituents; or [0251]
R.sub.6 and R.sub.6', together with the nitrogen atom to which they
are bound, may optionally form a (C3-C10)-heterocyclic ring system
wherein said ring system may be optionally substituted with up to 3
substituents independently selected from J.
DEFINITIONS
[0252] The term "aryl" as used herein means a monocyclic or
bicyclic carbocyclic aromatic ring system. Phenyl is an example of
a monocyclic aromatic ring system. Bicyclic aromatic ring systems
include systems wherein both rings are aromatic, e.g., naphthyl,
and systems wherein only one of the two rings is aromatic, e.g.,
tetralin. It is understood that as used herein, the term
"(C6-C10)-aryl-" includes any one of a C6, C7, C8, C9, and C10
monocyclic or bicyclic carbocyclic aromatic ring.
[0253] The term "heterocyclyl" as used herein means a monocyclic or
bicyclic non-aromatic ring system having 1 to 3 heteroatom or
heteroatom groups in each ring selected from O, N, NH, S, SO, and
SO.sub.2 in a chemically stable arrangement. In a bicyclic
non-aromatic ring system embodiment of "heterocyclyl" one or both
rings may contain said heteroatom or heteroatom groups. It is
understood that as used herein, the term "(C5-C10)-heterocyclyl-"
includes any one of a C5, C6, C7, C8, C9, and C10 monocyclic or
bicyclic non-aromatic ring system having 1 to 3 heteroatom or
heteroatom groups in each ring selected from O, N, NH, and S in a
chemically stable arrangement.
[0254] The term "heteroaryl" as used herein means a monocyclic or
bicyclic aromatic ring system having 1 to 3 heteroatom or
heteroatom groups in each ring selected from O, N, NH, and S in a
chemically stable arrangement. In such a bicyclic aromatic ring
system embodiment of "heteroaryl":
[0255] one or both rings may be aromatic; and
[0256] one or both rings may contain said heteroatom or heteroatom
groups. It is understood that as used herein, the term
"(C5-C10)-heteroaryl-" includes any one of a C5, C6, C7, C8, C9,
and C10 monocyclic or bicyclic aromatic ring system having 1 to 3
heteroatom or heteroatom groups in each ring selected from O, N,
NH, and S in a chemically stable arrangement.
[0257] The term "aliphatic" as used herein means a straight chained
or branched alkyl, alkenyl or alkynyl. It is understood that as
used herein, the term "(C1-C12)-aliphatic-" includes any one of a
C1, C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, and C12 straight or
branched alkyl chain of carbon atoms. It is also understood that
alkenyl or alkynyl embodiments need at least two carbon atoms in
the aliphatic chain. The term "cycloalkyl or cycloalkenyl" refers
to a monocyclic or fused or bridged bicyclic carbocyclic ring
system that is not aromatic. Cycloalkenyl rings have one or more
units of unsaturation. It is also understood that as used herein,
the term "(C3-C10)-cycloalkyl- or -cycloalkenyl-" includes any one
of a C3, C4, C5, C6, C7, C8, C9, and C10 monocyclic or fused or
bridged bicyclic carbocyclic ring. Preferred cycloalkyl groups
include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
cyclohexenyl, cycloheptyl, cycloheptenyl, norbornyl, adamantyl and
decalin-yl.
[0258] The phrase "chemically stable arrangement" as used herein
refers to a compound structure that renders the compound
sufficiently stable to allow manufacture and administration to a
mammal by methods known in the art. Typically, such compounds are
stable at a temperature of 40.degree. C. or less, in the absence of
moisture or other chemically reactive condition, for at least a
week.
EMBODIMENTS
[0259] According to one embodiment of compounds of formula I,
the
##STR00006##
radical is,
##STR00007##
wherein; in R', R.sub.10, and R.sub.10', X and Y are both a bond
and Z is hydrogen; and in R.sub.9'; X is a bond; Y is a bond,
--CH.sub.2--, or --C(O)--; and Z is (C1-C12)-aliphatic-, [0260]
(C3-C10)-cycloalkyl- or -cycloalkenyl-, [0261] [(C3-C10)-cycloalkyl
or -cycloalkenyl]-(C1-C12)-aliphatic-, [0262] (C6-C10)-aryl-,
[0263] (C6-C10)-aryl-(C1-C12)aliphatic-, [0264]
(C3-C10)-heterocyclyl-, [0265]
(C3-C10)-heterocyclyl-(C1-C12)aliphatic-, [0266]
(C5-C10)-heteroaryl-, or [0267]
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; [0268] wherein up to three
aliphatic carbon atoms in Z may be optionally replaced with S,
--S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)--, in a
chemically stable arrangement; [0269] wherein any ring may be
optionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,
(C3-C10)cycloalkyl, or (C3-C10)heterocyclyl; [0270] wherein Z may
be independently and optionally substituted with up to 3
substituents independently selected from J.
[0271] According to another embodiment, in R.sub.9';
X is a bond; Y is a bond; and Z is (C1-C12)-aliphatic-, [0272]
(C3-C10)-cycloalkyl- or -cycloalkenyl-, [0273] [(C3-C10)-cycloalkyl
or -cycloalkenyl]-(C1-C12)-aliphatic-, [0274] (C6-C10)-aryl-,
[0275] (C6-C10)-aryl-(C1-C12)aliphatic-, [0276]
(C5-C10)-heteroaryl-, or [0277]
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; [0278] wherein up to three
aliphatic carbon atoms in Z may be optionally replaced with S,
--S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)--, in a
chemically stable arrangement; [0279] wherein any ring may be
optionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,
(C3-C10)cycloalkyl, or (C3-C10)heterocyclyl; [0280] wherein Z may
be independently and optionally substituted with up to 3
substituents independently selected from J.
[0281] According to another embodiment, in R.sub.9';
X is a bond; Y is a bond; and Z is (C1-C12)-aliphatic-, [0282]
(C3-C10)-cycloalkyl- or -cycloalkenyl-, [0283] [(C3-C10)-cycloalkyl
or -cycloalkenyl]-(C1-C12)-aliphatic-, or [0284]
(C6-C10)-aryl-(C1-C12)aliphatic-, [0285] wherein up to three
aliphatic carbon atoms in Z may be optionally replaced with S,
--S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)--, in a
chemically stable arrangement; [0286] wherein Z may be
independently and optionally substituted with up to 3 substituents
independently selected from J.
[0287] According to another embodiment, R.sub.9' is
##STR00008##
[0288] According to another embodiment, R.sub.9' is
##STR00009##
[0289] According to another embodiment, R.sub.9' is ethyl.
[0290] According to another embodiment of compounds of formula I,
in R.sub.9, R.sub.10, and R.sub.10', X and Y are both a bond and Z
is hydrogen; and in R.sub.9';
X is a bond;
Y is --C(O)--; and
[0291] Z is (C1-C12)-aliphatic-, or [0292]
(C3-C10)-heterocyclyl-(C1-C12)aliphatic-; [0293] wherein up to
three aliphatic carbon atoms in Z may be optionally replaced with
S, --S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)--, in a
chemically stable arrangement; [0294] wherein any ring may be
optionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,
(C3-C10)cycloalkyl, or (C3-C10)heterocyclyl; [0295] wherein Z may
be independently and optionally substituted with up to 3
substituents independently selected from J.
[0296] According to another embodiment, Z is --O--(C1-C6)-aliphatic
or --N(R').sub.2, wherein the two R' groups bound to the nitrogen
atom may optionally form a 3- to 7-membered saturated or partially
unsaturated ring system wherein up to 3 ring atoms may be
optionally replaced with a heteroatom independently selected from
N, NH, O, S, SO, and SO.sub.2, wherein said ring system may be
optionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,
(C3-C10)cycloalkyl, or a (C3-C10)heterocyclyl, wherein any ring has
up to 3 substituents selected independently from J.
[0297] According to another embodiment of compounds of formula I, Z
is --N(R').sub.2, wherein the two R' groups bound to the nitrogen
atom may optionally form a 3- to 7-membered saturated or partially
unsaturated ring system wherein up to 3 ring atoms may be
optionally replaced with a heteroatom independently selected from
N, NH, O, S, SO, and SO.sub.2, wherein said ring system may be
optionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,
(C3-C10)cycloalkyl, or a (C3-C10)heterocyclyl, wherein any ring has
up to 3 substituents selected independently from J.
[0298] According to another embodiment of compounds of formula I,
in R.sub.9, and R.sub.10, X and Y are both a bond and Z is
hydrogen; and in each of R.sub.9' and R.sub.10' independently,
X is a bond; Y is a bond; and Z is (C1-C12)-aliphatic-, [0299]
(C3-C10)-cycloalkyl- or -cycloalkenyl-, [0300] [(C3-C10)-cycloalkyl
or -cycloalkenyl]-(C1-C12)-aliphatic-, [0301] (C6-C10)-aryl-,
[0302] (C6-C10)-aryl-(C1-C12)aliphatic-, [0303]
(C3-C10)-heterocyclyl-, [0304]
(C3-C10)-heterocyclyl-(C1-C12)aliphatic-, [0305]
(C5-C10)-heteroaryl-, or [0306]
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-; [0307] wherein up to three
aliphatic carbon atoms in Z may be optionally replaced with S,
--S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)--, in a
chemically stable arrangement; [0308] wherein any ring may be
optionally fused to a (C6-C10)aryl, (C5-C10)heteroaryl,
(C3-C10)cycloalkyl, or (C3-C10)heterocyclyl; [0309] wherein Z may
be independently and optionally substituted with up to 3
substituents independently selected from J.
[0310] According to another embodiment, Z in each of R.sub.9' and
R.sub.10' independently, is [0311] (C1-C12)-aliphatic-, [0312]
(C3-C10)-cycloalkyl- or -cycloalkenyl-, or [0313]
[(C3-C10)-cycloalkyl or -cycloalkenyl]-(C1-C12)-aliphatic-; [0314]
wherein up to three aliphatic carbon atoms in Z may be optionally
replaced with S, --S(O)--, --S(O).sub.2--, --O--, --N--, or
--N(H)--, in a chemically stable arrangement; [0315] wherein Z may
be independently and optionally substituted with up to 3
substituents independently selected from J.
[0316] According to another embodiment Z, in each of R.sub.9', and
R.sub.10', independently is (C1-C6)-aliphatic-.
[0317] According to another embodiment of compounds of formula I,
in R.sub.10, and R.sub.10', X and Y are both a bond and Z is
hydrogen; and in R.sub.9 and R.sub.9';
X is a bond, Y is a bond, and Z is (C1-C6)-aliphatic-, [0318]
wherein Z may be independently and optionally substituted with up
to 3 substituents independently selected from J.
[0319] According to another embodiment of compounds of formula I, W
is:
##STR00010##
wherein in the W, the NR.sub.6R.sub.6 is selected from --NH--
(C1-C6 aliphatic), --NH-- (C3-C6 cycloalkyl),
--NH--CH(CH.sub.3)-aryl, or --NH--CH(CH.sub.3)-heteroaryl, wherein
said aryl or said heteroaryl is optionally substituted with up to 3
halogens.
[0320] According to another embodiment in the W, the
NR.sub.6R.sub.6 is:
##STR00011##
[0321] According to another embodiment in the W, the
NR.sub.6R.sub.6 is:
##STR00012##
[0322] According to another embodiment in the W, the
NR.sub.6R.sub.6 is:
##STR00013##
[0323] According to another embodiment in the W, the
NR.sub.6R.sub.6 is:
##STR00014##
[0324] According to another embodiment in compounds of formula I,
the NR.sub.6R.sub.6 in the W radical is:
##STR00015##
[0325] According to another embodiment, the NR.sub.6R.sub.6 in the
W radical is:
##STR00016##
[0326] According to another embodiment, in the W, the
NR.sub.6R.sub.6 is:
##STR00017##
[0327] According to another embodiment W in compounds of formula I
is:
##STR00018##
wherein R.sub.8 is as defined above.
[0328] According to another embodiment each R.sub.8 together with
the boron atom, is a (C5-C10)-membered heterocyclic ring having no
additional heteroatoms other than the boron and the two oxygen
atoms.
[0329] In another embodiment W is:
##STR00019##
[0330] wherein R' is (C1-C6)-aliphatic.
[0331] In another embodiment R' is methyl.
[0332] According to another embodiment of compounds of formula I,
R.sub.5' is hydrogen and R.sub.5 is:
##STR00020##
[0333] According to another embodiment R.sub.5' is hydrogen and
R.sub.5 is:
##STR00021##
[0334] According to another embodiment in compounds of formula I,
R.sub.5' and R.sub.5 is:
##STR00022##
[0335] According to another embodiment of compounds of formula I,
R.sub.2, R.sub.4, and R.sub.7 are each independently H, methyl,
ethyl, or propyl.
[0336] According to another embodiment R.sub.2, R.sub.4, and
R.sub.7 are each hydrogen.
[0337] According to another embodiment of compounds of formula I,
R.sub.3 is:
##STR00023##
[0338] According to another embodiment R.sub.3 is:
##STR00024##
[0339] According to another embodiment R.sub.3 is:
##STR00025##
[0340] According to another embodiment of compounds of formula I,
R.sub.1 is:
##STR00026##
[0341] According to another embodiment R.sub.1 is:
##STR00027##
[0342] According to another embodiment wherein R.sub.1 is isopropyl
or cyclohexyl.
[0343] According to another embodiment of compounds of formula I,
the
##STR00028##
radical is:
##STR00029##
wherein:
[0344] R.sub.6, R.sub.6', R.sub.7, and R.sub.12, are as defined in
any of the embodiments herein.
[0345] According to another embodiment in the
##STR00030##
radical;
[0346] R.sub.6' and R.sub.7 are both hydrogen;
[0347] R.sub.6 is:
[0348] (C1-C12)-aliphatic-;
[0349] (C6-C10)-aryl-,
[0350] (C6-C10)-aryl-(C1-C12)aliphatic-,
[0351] (C3-C10)-cycloalkyl or -cycloalkenyl-,
[0352] [(C3-C10)-cycloalkyl or
-cycloalkenyl]-(C1-C12)-aliphatic-,
[0353] (C3-C10)-heterocyclyl-,
[0354] (C3-C10)-heterocyclyl-(C1-C12)-aliphatic-,
[0355] (C5-C10)-heteroaryl-, or
[0356] (C5-C10)-heteroaryl-(C1-C12)-aliphatic-; [0357] wherein up
to 3 aliphatic carbon atoms in R.sub.6 may be optionally replaced
by S, --S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)--, in a
chemically stable arrangement; and [0358] wherein R.sub.6 may be
optionally substituted with up to 3 substituents independently
selected from J; and R.sub.12 is as defined in any of the
embodiments herein.
[0359] According to another embodiment
R.sub.6 is:
[0360] (C1-C12)-aliphatic-;
[0361] (C6-C10)-aryl-(C1-C12)aliphatic-, or
[0362] (C3-C10)-cycloalkyl or -cycloalkenyl-; [0363] wherein up to
3 aliphatic carbon atoms in R.sub.6 may be optionally replaced by
S, --S(O)--, --S(O).sub.2--, --O--, --N--, or --N(H)--, in a
chemically stable arrangement; [0364] wherein R.sub.6 may be
optionally substituted with up to 3 substituents independently
selected from J; and R.sub.12 is as defined in any of the
embodiments herein.
[0365] According to another embodiment the radical is:
##STR00031##
[0366] According to another embodiment the
##STR00032##
radical is:
##STR00033##
[0367] According to another embodiment of compounds of formula I, V
is --C(O)-- and R is a bond.
[0368] According to another embodiment of compounds of formula I, V
is --C(O)--, R is a bond, and
T is:
[0369] (C3-C10)-heterocyclyl- or (C5-C10)heteroaryl-; [0370]
wherein each T is optionally substituted with up to 3 J
substituents.
[0371] According to another embodiment, T is
(C5-C6)heterocyclyl- or (C5-C6)heteroaryl-; [0372] wherein each T
is optionally substituted with up to 3 J substituents.
[0373] According to another embodiment, T is:
##STR00034##
wherein:
[0374] Z' is independently O, S, NR', or C(R').sub.2.
[0375] According to another embodiment, T is:
##STR00035##
[0376] According to another embodiment, this invention does not
include the following compounds: [0377] 1.
3-Acetyl-4,5-dimethyl-1H-pyrrole-2-carboxylic acid
(cyclohexyl-{1-[3-cyclohexyl-2-(1-cyclopropylaminooxalyl-butylcarbamoyl)--
pyrrolidine-1-carbonyl]-2,2-dimethyl-propylcarbamoyl}-methyl)-amide;
[0378] 2. 3-Acetyl-4,5-dimethyl-1H-pyrrole-2-carboxylic acid
(cyclohexyl-{1-[2-(1-cyclopropylaminooxalyl-butylcarbamoyl)-3-isopropyl-p-
yrrolidine-1-carbonyl]-2,2-dimethyl-propylcarbamoyl}-methyl)-amide;
[0379] 3. 3-Acetyl-4,5-dimethyl-1H-pyrrole-2-carboxylic acid
(cyclohexyl-{1-[2-(1-cyclopropylaminooxalyl-butylcarbamoyl)-4-(quinazolin-
-4-yloxy)-pyrrolidine-1-carbonyl]-2,2-dimethyl-propylcarbamoyl}-methyl)-am-
ide; and [0380] 4. 3-Acetyl-4,5-dimethyl-1H-pyrrole-2-carboxylic
acid
({1-[4-(5-chloro-pyridin-2-yloxy)-2-(1-cyclopropylaminooxalyl-butylcarbam-
oyl)-pyrrolidine-1-carbonyl]-2,2-dimethyl-propylcarbamoyl}-cyclohexyl-meth-
yl)-amide (e.g., compounds 63, 64, 66, and 67 of WO 03/087092).
[0381] According to yet another embodiment, this invention does not
include the following compounds
wherein: V is --C(O)--, R is a bond, T is the (C5-C10)-heteroaryl
3-acetyl-4,5-dimethyl-1H-pyrrole and the
##STR00036##
radical is:
##STR00037## ##STR00038## ##STR00039## ##STR00040##
##STR00041##
(e.g., substituted proline radicals at pages 56 and 57 of WO
03/087092).
[0382] According to another embodiment, this invention does not
include compounds wherein:
V is --C(O)--;
[0383] R is a bond; and T is 3-acetyl-4,5-dimethyl-1H-pyrrole
(e.g., compounds of formula II'' at page 85 of WO 03/087092).
[0384] According to another embodiment, this invention does not
include compounds wherein T is a C5-heteroaryl (e.g., compounds of
formula II at page 22 of WO 03/087092).
[0385] According to another embodiment, this invention does not
include compounds wherein T is an optionally substituted pyrrole
group (e.g., compounds of formula II at page 22 of WO
03/087092).
[0386] According to another embodiment, this invention does not
include compounds wherein:
V is --C(O)--, --S(O)--, or --S(O).sub.2--;
[0387] R is a bond; and
T is:
##STR00042##
[0388] wherein:
[0389] R.sub.14 is --H, --S(O)R', --S(O).sub.2R', --C(O)R',
--C(O)OR', --C(O)N(R').sub.2, --N(R')C(O)R', --N(COR')COR',
--SO.sub.2N(R').sub.2, --SO.sub.3R', --C(O)C(O)R',
--C(O)CH.sub.2C(O)R', --C(S)R', --C(S)N(R').sub.2,
--(CH.sub.2).sub.0-2NHC(O)R', --N(R')N(R')COR',
--N(R')N(R')C(O)OR', --N(R')N(R')CON(R').sub.2, --N(R')SO.sub.2R',
--N(R') SO.sub.2N(R').sub.2, --N(R')C(O)OR', --N(R')C(O)R',
--N(R')C(S)R', --N(R')C(O)N(R').sub.2, --N(R')C(S)N(R').sub.2,
--N(COR')COR', --N(OR')R', --C(.dbd.NH)N(R').sub.2, --C(O)N(OR')R',
--C(.dbd.NOR')R', --OP(O)(OR').sub.2, --P(O)(R').sub.2,
--P(O)(OR').sub.2, or --P(O)(H)(OR');
[0390] R.sub.15 and R.sub.16 are independently halogen, --OR',
--OC(O)N(R').sub.2, --NO.sub.2, --CN, --CF.sub.3, --OCF.sub.3,
--R', oxo, 1,2-methylenedioxy, 1,2-ethylenedioxy, --N(R').sub.2,
--SR', --SOR', --SO.sub.2R', --SO.sub.2N(R').sub.2, --SO.sub.3R',
--C(O)R', --C(O)C(O)R', --C(O)CH.sub.2C(O)R', --C(S)R', --C(O)OR',
--OC(O)R', --C(O)N(R').sub.2, --OC(O)N(R').sub.2,
--C(S)N(R').sub.2, --(CH.sub.2).sub.0-2NHC(O)R', --N(R')N(R')COR',
--N(R')N(R')C(O)OR', --N(R')N(R')CON(R').sub.2, --N(R')SO.sub.2R',
--N(R')SO.sub.2N(R').sub.2, --N(R')C(O)OR', --N(R')C(O)R',
--N(R')C(S)R', --N(R')C(O)N(R').sub.2, --N(R')C(S)N(R').sub.2,
--N(COR')COR', --N(OR')R', --CN, --C(.dbd.NH)N(R').sub.2,
--C(O)N(OR')R', --C(.dbd.NOR')R', --OP(O)(OR').sub.2,
--P(O)(R').sub.2, --P(O)(OR').sub.2, or --P(O)(H)(OR');
[0391] Z.sub.2 is .dbd.O, .dbd.NR', .dbd.NOR', or
.dbd.C(R').sub.2;
[0392] R.sub.19 is --OR', --CF.sub.3, --OCF.sub.3, --R',
--N(R').sub.2, --SR', --C(O)R', --COOR', --CON(R').sub.2,
--N(R')COR', or --N(COR')COR'; wherein
[0393] two R' groups together with the atoms to which they are
bound form a 3- to 10-membered aromatic or non-aromatic ring having
up to 3 heteroatoms independently selected from N, NH, O, S, SO, or
SO.sub.2, wherein the ring is optionally fused to a (C6-C10)aryl,
(C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a (C3-C10)heterocyclyl,
and wherein any ring has up to 3 substituents selected
independently from J.sub.2; or
[0394] each R' is independently selected from: [0395] hydrogen-,
[0396] (C1-C12)-aliphatic-, [0397] (C3-C10)-cycloalkyl or
-cycloalkenyl-, [0398] [(C3-C10)-cycloalkyl or
-cycloalkenyl]-(C1-C12)-aliphatic-, [0399] (C6-C10)-aryl-, [0400]
(C6-C10)-aryl-(C1-C12)aliphatic-, [0401] (C3-C10)-heterocyclyl-,
[0402] (C6-C10)-heterocyclyl-(C1-C12)aliphatic-, [0403]
(C5-C10)-heteroaryl-, or [0404]
(C5-C10)-heteroaryl-(C1-C12)-aliphatic-, wherein R' has up to 3
substituents selected independently from J.sub.2; and
[0405] J.sub.2 is halogen, --OR', --OC(O)N(R').sub.2, --NO.sub.2,
--CN, --CF.sub.3, --OCF.sub.3, --R', oxo, thioxo,
1,2-methylenedioxy, --N(R').sub.2, --SR', --SOR', --SO.sub.2R',
--SO.sub.2N(R').sub.2, --SO.sub.3R', --C(O)R', --C(O)C(O)R',
--C(O)CH.sub.2C(O)R', --C(S)R', --C(O)OR', --OC(O)R',
--C(O)N(R').sub.2, --OC(O)N(R').sub.2, --C(S)N(R').sub.2,
--(CH.sub.2).sub.0-2NHC(O)R', --N(R')N(R')COR',
--N(R')N(R')C(O)OR', --N(R')N(R')CON(R').sub.2, --N(R')SO.sub.2R',
--N(R')SO.sub.2N(R').sub.2, --N(R')C(O)OR', --N(R')C(O)R',
--N(R')C(S)R', --N(R')C(O)N(R').sub.2, --N(R')C(S)N(R').sub.2,
--N(COR')COR', --N(OR')R', --CN, --C(.dbd.NH)N(R').sub.2,
--C(O)N(OR')R', --C(.dbd.NOR')R', --OP(O)(OR').sub.2,
--P(O)(R').sub.2, --P(O)(OR').sub.2, or --P(O)(H)(OR') (e.g.,
compounds of formula II at page 22 of WO 03/087092).
[0406] According to another preferred embodiment in compounds of
formula I, the compound is:
##STR00043## ##STR00044## ##STR00045## ##STR00046## ##STR00047##
##STR00048## ##STR00049## ##STR00050## ##STR00051## ##STR00052##
##STR00053## ##STR00054## ##STR00055## ##STR00056## ##STR00057##
##STR00058## ##STR00059## ##STR00060## ##STR00061##
[0407] The compounds of this invention may contain one or more
asymmetric carbon atoms and thus may occur as racemates and racemic
mixtures, single enantiomers, diastereomeric mixtures and
individual diastereomers. All such isomeric forms of these
compounds are expressly included in the present invention. Each
stereogenic carbon may be of the R or S configuration.
[0408] In another embodiment, the compounds of this invention have
the structure and stereochemistry depicted in compounds 1-77.
[0409] Any of the embodiments recited above, including those
embodiments in the above species, may be combined to produce a
preferred embodiment of this invention.
[0410] As can be appreciated by the skilled artisan, the synthetic
schemes shown are not intended to comprise a comprehensive list of
all means by which the compounds described and claimed in this
application may be synthesized. Other equivalent schemes, which
will be readily apparent to the ordinary skilled organic chemist,
may alternatively be used to synthesize various portions of the
molecule as illustrated by the general schemes below. Additionally,
the various synthetic steps described above may be performed in an
alternate sequence or order to give the desired compounds. Other
equivalent schemes, which will be readily apparent to the ordinary
skilled organic chemist, may alternatively be used to synthesize
various portions of the molecule as illustrated by the general
schemes below, and the preparative examples that follow.
[0411] Abbreviations which are used in the schemes, preparations
and the examples that follow are: [0412] DCM: dichloromethane
[0413] THF: tetrahydrofuran [0414] DMF: N,N,-dimethylformamide
[0415] EtOAc: ethyl acetate [0416] AcOH: acetic acid [0417] NMM:
N-methylmorpholine [0418] NMP: N-methylpyrrolidinone [0419] EtOH:
ethanol [0420] t-BuOH: tert-butanol [0421] Et.sub.2O: diethyl ether
[0422] DMSO: dimethyl sulfoxide [0423] DCCA: dichloroacetic acid
[0424] DIEA: diisopropylethylamine [0425] MeCN: acetonitrile [0426]
TFA: trifluoroacetic acid [0427] DBU:
1,8-diazabicyclo[5.4.0]undec-7-ene [0428] DEAD: diethyl
azodicarboxylate [0429] HOBt: 1-hydroxybenzotriazole hydrate [0430]
HOAt: 1-hydroxy-7-azabenzotriazole [0431] EDC:
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride [0432]
Boc: tert-butyloxycarbonyl [0433] Boc.sub.2O:
di-tert-butyldicarbonate [0434] Cbz: benzyloxycarbonyl [0435]
Cbz-Cl: benzyl chloroformate [0436] Fmoc: 9-fluorenyl
methyloxycarbonyl [0437] SEM: silylethoxymethyl [0438] TBAF:
tetrabutylammonium fluoride [0439] Chg: cyclohexylglycine [0440]
t-BG: tert-butylglycine [0441] mCBPA: 3-chloroperoxybenzoic acid
[0442] DAST: (diethylamino)sulfur trifluoride [0443] TEMPO:
2,2,6,6-tetramethyl-1-piperidinyloxy, free radical [0444] PyBOP:
tris(pyrrolidino)bromophosphonium hexafluorophosphate [0445] TBTU
or HATU: 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium
tetrafluoroborate [0446] DMAP: 4-dimethylaminopyridine [0447] AIBN:
2,2'-azobisisobutyronitrile [0448] rt or RT: room temperature
[0449] ON: overnight [0450] ND: not determined [0451] MS: mass
spectrometry [0452] LC: liquid chromatography
General Synthetic Methodology:
[0453] The compounds of this invention may be prepared in general
by methods known to those skilled in the art. Schemes 1-6 below
illustrate synthetic routes to the compounds of the present
invention. Other equivalent schemes, which will be readily apparent
to the ordinary skilled organic chemist, may alternatively be used
to synthesize various portions of the molecule as illustrated by
the general schemes below, and the preparative examples that
follow.
##STR00062##
##STR00063##
##STR00064##
##STR00065##
##STR00066##
##STR00067##
[0454] Scheme 1-6 above provide synthetic pathways for the
preparation of the compounds of this invention. Many of the
starting proline derivatives may be purchased commercially from
chemical suppliers known to those in the art. Intermediate A1 may
be prepared according to the procedure described in J. Med. Chem.
39, p. 2367 (1996).
[0455] Although certain embodiments are depicted and described
below, it will be appreciated that compounds of this invention can
be prepared according to the methods described generally above
using appropriate starting materials generally available to one of
ordinary skill in the art.
[0456] Another embodiment of this invention provides a
pharmaceutical composition comprising a compound of formula I or a
pharmaceutically acceptable salt thereof. According to another
embodiment, the compound of formula I is present in an amount
effective to decrease the viral load in a sample or in a patient,
wherein said virus encodes a serine protease necessary for the
viral life cycle, and a pharmaceutically acceptable carrier.
[0457] If pharmaceutically acceptable salts of the compounds of
this invention are utilized in these compositions, those salts are
preferably derived from inorganic or organic acids and bases.
Included among such acid salts are the following: acetate, adipate,
alginate, aspartate, benzoate, benzene sulfonate, bisulfate,
butyrate, citrate, camphorate, camphor sulfonate,
cyclopentane-propionate, digluconate, dodecylsulfate,
ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate,
hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide,
hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate,
methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate,
pamoate, pectinate, persulfate, 3-phenyl-propionate, picrate,
pivalate, propionate, succinate, tartrate, thiocyanate, tosylate
and undecanoate. Base salts include ammonium salts, alkali metal
salts, such as sodium and potassium salts, alkaline earth metal
salts, such as calcium and magnesium salts, salts with organic
bases, such as dicyclohexylamine salts, N-methyl-D-glucamine, and
salts with amino acids such as arginine, lysine, and so forth.
[0458] Also, the basic nitrogen-containing groups may be
quaternized with such agents as lower alkyl halides, such as
methyl, ethyl, propyl, and butyl chloride, bromides and iodides;
dialkyl sulfates, such as dimethyl, diethyl, dibutyl and diamyl
sulfates, long chain halides such as decyl, lauryl, myristyl and
stearyl chlorides, bromides and iodides, aralkyl halides, such as
benzyl and phenethyl bromides and others. Water or oil-soluble or
dispersible products are thereby obtained.
[0459] The compounds utilized in the compositions and methods of
this invention may also be modified by appending appropriate
functionalities to enhance selective biological properties. Such
modifications are known in the art and include those which increase
biological penetration into a given biological system (e.g., blood,
lymphatic system, central nervous system), increase oral
availability, increase solubility to allow administration by
injection, alter metabolism and alter rate of excretion.
[0460] Pharmaceutically acceptable carriers that may be used in
these compositions include, but are not limited to, ion exchangers,
alumina, aluminum stearate, lecithin, serum proteins, such as human
serum albumin, buffer substances such as phosphates, glycine,
sorbic acid, potassium sorbate, partial glyceride mixtures of
saturated vegetable fatty acids, water, salts or electrolytes, such
as protamine sulfate, disodium hydrogen phosphate, potassium
hydrogen phosphate, sodium chloride, zinc salts, colloidal silica,
magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based
substances, polyethylene glycol, sodium carboxymethylcellulose,
polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers,
polyethylene glycol and wool fat.
[0461] According to another embodiment, the compositions of this
invention are formulated for pharmaceutical administration to a
mammal. In another embodiment the compositions of this invention
are formulated for pharmaceutical administration to a human
being.
[0462] Such pharmaceutical compositions of the present invention
may be administered orally, parenterally, by inhalation spray,
topically, rectally, nasally, buccally, vaginally or via an
implanted reservoir. The term "parenteral" as used herein includes
subcutaneous, intravenous, intramuscular, intra-articular,
intra-synovial, intrasternal, intrathecal, intrahepatic,
intralesional and intracranial injection or infusion techniques. In
another embodiment, the compositions are administered orally or
intravenously.
[0463] Sterile injectable forms of the compositions of this
invention may be aqueous or oleaginous suspension. These
suspensions may be formulated according to techniques known in the
art using suitable dispersing or wetting agents and suspending
agents. The sterile injectable preparation may also be a sterile
injectable solution or suspension in a non-toxic parenterally
acceptable diluent or solvent, for example as a solution in
1,3-butanediol. Among the acceptable vehicles and solvents that may
be employed are water, Ringer's solution and isotonic sodium
chloride solution. In addition, sterile, fixed oils are
conventionally employed as a solvent or suspending medium. For this
purpose, any bland fixed oil may be employed including synthetic
mono- or di-glycerides. Fatty acids, such as oleic acid and its
glyceride derivatives are useful in the preparation of injectables,
as are natural pharmaceutically-acceptable oils, such as olive oil
or castor oil, especially in their polyoxyethylated versions. These
oil solutions or suspensions may also contain a long-chain alcohol
diluent or dispersant, such as carboxymethyl cellulose or similar
dispersing agents which are commonly used in the formulation of
pharmaceutically acceptable dosage forms including emulsions and
suspensions. Other commonly used surfactants, such as Tweens, Spans
and other emulsifying agents or bioavailability enhancers which are
commonly used in the manufacture of pharmaceutically acceptable
solid, liquid, or other dosage forms may also be used for the
purposes of formulation.
[0464] Dosage levels of between about 0.01 and about 100 mg/kg body
weight per day, preferably between about 0.5 and about 75 mg/kg
body weight per day of the protease inhibitor compounds described
herein are useful in a monotherapy for the prevention and treatment
of antiviral, particularly anti-HCV mediated disease. Typically,
the pharmaceutical compositions of this invention will be
administered from about 1 to about 5 times per day or
alternatively, as a continuous infusion. Such administration can be
used as a chronic or acute therapy. The amount of active ingredient
that may be combined with the carrier materials to produce a single
dosage form will vary depending upon the host treated and the
particular mode of administration. A typical preparation will
contain from about 5% to about 95% active compound (w/w). In
another embodiment, such preparations contain from about 20% to
about 80% active compound.
[0465] When the compositions of this invention comprise a
combination of a compound of formula I and one or more additional
therapeutic or prophylactic agents, both the compound and the
additional agent should be present at dosage levels of between
about 10 to 100% and in another embodiment between about 10 to 80%
of the dosage normally administered in a monotherapy regimen.
[0466] The pharmaceutical compositions of this invention may be
orally administered in any orally acceptable dosage form including,
but not limited to, capsules, tablets, aqueous suspensions or
solutions. In the case of tablets for oral use, carriers that are
commonly used include lactose and corn starch. Lubricating agents,
such as magnesium stearate, are also typically added. For oral
administration in a capsule form, useful diluents include lactose
and dried cornstarch. When aqueous suspensions are required for
oral use, the active ingredient is combined with emulsifying and
suspending agents. If desired, certain sweetening, flavoring or
coloring agents may also be added.
[0467] Alternatively, the pharmaceutical compositions of this
invention may be administered in the form of suppositories for
rectal administration. These may be prepared by mixing the agent
with a suitable non-irritating excipient which is solid at room
temperature but liquid at rectal temperature and therefore will
melt in the rectum to release the drug. Such materials include
cocoa butter, beeswax and polyethylene glycols.
[0468] The pharmaceutical compositions of this invention may also
be administered topically, especially when the target of treatment
includes areas or organs readily accessible by topical application,
including diseases of the eye, the skin, or the lower intestinal
tract. Suitable topical formulations are readily prepared for each
of these areas or organs.
[0469] Topical application for the lower intestinal tract may be
effected in a rectal suppository formulation (see above) or in a
suitable enema formulation. Topically-transdermal patches may also
be used.
[0470] For topical applications, the pharmaceutical compositions
may be formulated in a suitable ointment containing the active
component suspended or dissolved in one or more carriers. Carriers
for topical administration of the compounds of this invention
include, but are not limited to, mineral oil, liquid petrolatum,
white petrolatum, propylene glycol, polyoxyethylene,
polyoxypropylene compound, emulsifying wax and water.
Alternatively, the pharmaceutical compositions may be formulated in
a suitable lotion or cream containing the active components
suspended or dissolved in one or more pharmaceutically acceptable
carriers. Suitable carriers include, but are not limited to,
mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters
wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and
water.
[0471] For ophthalmic use, the pharmaceutical compositions may be
formulated as micronized suspensions in isotonic, pH adjusted
sterile saline, or, preferably, as solutions in isotonic, pH
adjusted sterile saline, either with our without a preservative
such as benzylalkonium chloride. Alternatively, for ophthalmic
uses, the pharmaceutical compositions may be formulated in an
ointment such as petrolatum.
[0472] The pharmaceutical compositions of this invention may also
be administered by nasal aerosol or inhalation. Such compositions
are prepared according to techniques well known in the art of
pharmaceutical formulation and may be prepared as solutions in
saline, employing benzyl alcohol or other suitable preservatives,
absorption promoters to enhance bioavailability, fluorocarbons,
and/or other conventional solubilizing or dispersing agents.
[0473] In another embodiment, the pharmaceutical compositions are
formulated for oral administration.
[0474] In one embodiment, the compositions of this invention
additionally comprise another agent, including a cytochrome P-450
inhibitor. Such cytochrome P-450 inhibitors include, but are not
limited to, ritonavir.
[0475] In another embodiment, the compositions of this invention
additionally comprise another anti-viral agent, including an
anti-HCV agent. Such anti-viral agents include, but are not limited
to, immunomodulatory agents, such as .alpha.-, .beta.-, and
.gamma.-interferons, pegylated derivatized interferon-.alpha.
compounds, and thymosin; other anti-viral agents, such as
ribavirin, amantadine, and telbivudine; other inhibitors of
hepatitis C proteases (NS2-NS3 inhibitors and NS3-NS4A inhibitors);
inhibitors of other targets in the HCV life cycle, including but
not limited to, helicase and polymerase inhibitors; inhibitors of
internal ribosome entry; broad-spectrum viral inhibitors, such as
IMPDH inhibitors (e.g., VX-497 and other IMPDH inhibitors disclosed
in U.S. Pat. Nos. 5,807,876 and 6,498,178, mycophenolic acid and
derivatives thereof); inhibitors of cytochrome P-450, such as
ritonavir, or combinations of any of the above.
[0476] Upon improvement of a patient's condition, a maintenance
dose of a compound, composition or combination of this invention
may be administered, if necessary. Subsequently, the dosage or
frequency of administration, or both, may be reduced, as a function
of the symptoms, to a level at which the improved condition is
retained when the symptoms have been alleviated to the desired
level, treatment should cease. Patients may, however, require
intermittent treatment on a long-term basis upon any recurrence of
disease symptoms.
[0477] It should also be understood that a specific dosage and
treatment regimen for any particular patient will depend upon a
variety of factors, including the activity of the specific compound
employed, the age, body weight, general health, sex, diet, time of
administration, rate of excretion, drug combination, and the
judgment of the treating physician and the severity of the
particular disease being treated. The amount of active ingredients
will also depend upon the particular described compound and the
presence or absence and the nature of the additional anti-viral
agent in the composition.
[0478] According to another embodiment, the invention provides a
method for treating a patient infected with a virus characterized
by a virally encoded serine protease that is necessary for the life
cycle of the virus by administering to said patient a
pharmaceutically acceptable composition of this invention. In
another embodiment, the methods of this invention are used to treat
a patient suffering from a HCV infection. Such treatment may
completely eradicate the viral infection or reduce the severity
thereof. In another embodiment, the methods of this invention are
used to treat a patient suffering from a HCV infection wherein the
patient is a human being.
[0479] In an alternate embodiment, the methods of this invention
additionally comprise the step of administering to said patient an
anti-viral agent preferably an anti-HCV agent. Such anti-viral
agents include, but are not limited to, immunomodulatory agents,
such as .alpha.-, .beta.-, and .gamma.-interferons, pegylated
derivatized interferon-.alpha. compounds, and thymosin; other
anti-viral agents, such as ribavirin, amantadine, and telbivudine;
other inhibitors of hepatitis C proteases (NS2-NS3 inhibitors and
NS3-NS4A inhibitors); inhibitors of other targets in the HCV life
cycle, including helicase and polymerase inhibitors; inhibitors of
internal ribosome entry; broad-spectrum viral inhibitors, such as
IMPDH inhibitors (e.g., VX-497 and other IMPDH inhibitors disclosed
in U.S. Pat. Nos. 5,807,876 and 6,498,178, mycophenolic acid and
derivatives thereof); inhibitors of cytochrome P-450, such as
ritonavir, or combinations of any of the above.
[0480] Such additional agent may be administered to said patient as
part of a single dosage form comprising both a compound of this
invention and an additional anti-viral agent. Alternatively the
additional agent may be administered separately from the compound
of this invention, as part of a multiple dosage form, wherein said
additional agent is administered prior to, together with or
following a composition comprising a compound of this
invention.
[0481] In yet another embodiment the present invention provides a
method of pre-treating a biological substance intended for
administration to a patient comprising the step of contacting said
biological substance with a pharmaceutically acceptable composition
comprising a compound of this invention. Such biological substances
include, but are not limited to, blood and components thereof such
as plasma, platelets, subpopulations of blood cells and the like;
organs such as kidney, liver, heart, lung, etc; sperm and ova; bone
marrow and components thereof, and other fluids to be infused into
a patient such as saline, dextrose, etc.
[0482] According to another embodiment the invention provides
methods of treating materials that may potentially come into
contact with a virus characterized by a virally encoded serine
protease necessary for its life cycle. This method comprises the
step of contacting said material with a compound according to the
invention. Such materials include, but are not limited to, surgical
instruments and garments (e.g. clothes, gloves, aprons, gowns,
masks, eyeglasses, footwear, etc.); laboratory instruments and
garments (e.g. clothes, gloves, aprons, gowns, masks, eyeglasses,
footwear, etc.); blood collection apparatuses and materials; and
invasive devices, such as shunts, stents, etc.
[0483] In another embodiment, the compounds of this invention may
be used as laboratory tools to aid in the isolation of a virally
encoded serine protease. This method comprises the steps of
providing a compound of this invention attached to a solid support;
contacting said solid support with a sample containing a viral
serine protease under conditions that cause said protease to bind
to said solid support; and eluting said serine protease from said
solid support. In another embodiment, the viral serine protease
isolated by this method is HCV NS3-NS4A protease.
[0484] In order that this invention be more fully understood, the
following preparative and testing examples are set forth. These
examples are for the purpose of illustration only and are not to be
construed as limiting the scope of the invention in any way.
EXAMPLES
[0485] .sup.1H-NMR spectra were recorded at 500 MHz using a Bruker
AMX 500 instrument. Mass spec. samples were analyzed on a MicroMass
ZQ or Quattro II mass spectrometer operated in single MS mode with
electrospray ionization. Samples were introduced into the mass
spectrometer using flow injection (FIA) or chromatography. Mobile
phase for all mass spec. analysis consisted of acetonitrile-water
mixtures with 0.2% formic acid as a modifier.
[0486] As used herein, the term "R.sub.t(min)" refers to the HPLC
retention time, in minutes, associated with the compound. The HPLC
retention times listed were either obtained from the mass spec.
data or using the following method: [0487] Instrument: Hewlett
Packard HP-1050; [0488] Column: YMC C18 (Cat. No. 326289C46);
[0489] Gradient/Gradient Time: 10-90% CH.sub.3CN/H2O over 9
minutes, then 100% CH.sub.3CN for 2 minutes; [0490] Flow Rate: 0.8
ml/min; [0491] Detector Wavelength: 215 nM and 245 nM.
[0492] Chemical naming for selected compounds herein was
accomplished using the naming program provided by CambridgeSoft
Corporations ChemDraw Ultra.RTM., version 7.0.1.
Example 1
Pyrazine-2-carboxylic acid
(cyclohexyl-{1-[2-(1-cyclopropylaminooxalyl-butylcarbamoyl)-3-isopropyl-p-
yrrolidine-1-carbonyl]-2,2-dimethyl-propylcarbamoyl}-methyl)-amide
(56)
[0493] To a stirring suspension of copper bromide-dimethylsulfide
(9.1 g, 44.28 mmol) in 100 mL of dry ether at -20.degree. C. was
added isopropenyl magnesium bromide 0.09M (100 mL). After 15 min.
of stirring, the temperature was lowered to -78.degree. C. and
enone 4a (4.0 g, 8.86 mmol, prepared according to procedure in
JACS, 117, p. 10775, (1995)) in 50 mL of ether was added followed
by TMSCl (2.25 mL, 18 mmol). The reaction mixture was stirred at
-78.degree. C. for 1 h and quenched with 100 mL of ammonium
hydroxide-ammonium chloride solution (1:4). Extracted with ether
and the organic phase was washed to remove all the copper salts.
The ether layer was dried with sodium sulfate and concentrated in
vacuo to an oil that was subjected to flash chromatography
(ether-hexanes (2:3) to provide 3.5 g (73%) of the desired
intermediate olefin.
[0494] .sup.1H NMR (CDCl.sub.3) .delta. 4.8 (d, 2H); 3.8 (m, 2H);
3.7 (d, 1H); 2.8 (m, 2H); 2.2 (d, 1H); 1.7 (s, 3H); 1.5 (s, 9H);
0.8 (s, 9H); 0.1 (s, 3H); 0.08 (s, 3H) ppm.
[0495] Hydrogenation with 10% Pd--C under 1 atmosphere of hydrogen
provided 3.5 g (100%) of the desired proline 5b.
[0496] HCl gas was bubbled 5 minutes to a solution of 5b (3.5 g,
6.47 mmol) in 50 mL of ethyl acetate at -20.degree. C. Stirred at
-20.degree. C. for 30 minutes then warmed up at rt and stirred for
1 h. It was concentrated in vacuo to 1.71 g (100%)of an oil that
was reduced with 2.5 equivalent of a 1M LAH in THF solution under
reflux for 4 h. Cooled and subjected to a Fieser work up which
provided 1.35 g (85%) of the desired compound 8b. .sup.1H NMR
(CDCl.sub.3) .delta. 4.0 (dd, 1H); 3.6 (m, 1H); 3.4 (m, 1H); 3.3
(m, 1H); 3.2 (m, 1H); 2.2 (m, 1H); 1.8 (m, 3H); 1.0 (d, 3H); 0.9
(d, 3H) ppm.
[0497] To a solution of potassium carbonate (190 mg, 1.38 mmol) in
4 mL of water at rt with stirring, was added 8b (357 mg, 2.5 mmol)
in 5 mL of THF. The solution was cooled to -2.degree. C. and Cbz
chloride (0.447 mL, 3.13 mmol) was added dropwise maintaining the
temperature at 0 to -2.degree. C. It was stirred for an additional
15 minutes, poured into water-ice. The aqueous phase was saturated
with salt and the organic phase separated. Further extraction with
ethyl acetate was necessary to extract all the compound. The
combined organics were washed with HCl 5%, water and brine, dried
with sodium sulfate and concentrated in vacuo to 416 mg (60%) on
benzoylated hydroxymethylpyrrolidine intermediate. 328 mg of this
material was oxidized with Jones reagent to provide 260 mg (75%) of
the proline intermediate. The above proline (260 mg, 0.889 mmol)
was esterified with isobutylene in dichloromethane with a catalytic
amount of concentrated sulfuric acid at rt in a seal vessel for 48
h to provide 289 mg (96%) of the intermediate ester. .sup.1H NMR
(CDCl.sub.3) .delta. 7.5 (m, 5H); 5.1 (m, 2H); 4.1 (dd, 1H); 3.6
(m, 1H); 3.5 (m, 1H); 2.1 (m, 2H); 1.7 (m, 2H); 1.5 (s, 9H); 1.1
(d, 3H); 1.0 (d, 3H) ppm.
[0498] Hydrogenation with 10% Pd/C in ethyl acetate gave 290 mg
(100%) of the desired compound 9b.
[0499] To a solution of Cbz-tert-butyl glycine (271 mg, 1.02 mmol)
in 2 mL of DCM at 0.degree. C. was added EDC (235 mg, 1.23 mmol),
HOBt (203 mg, 1.33 mmol) and DIEA (0.534 mL, 3.07 mmol). The
resulting mixture was stirred at 0.degree. C. for 15 min. after
which, the above amino ester 9b was slowly added in 2 mL of DCM.
The resulting reaction mixture was stirred at rt for 16 h.
Concentrated to a residue that was redissolved in EtOAc. Successive
washes with 0.5N HCL, satd' aqueous NaHCO.sub.3 and brine gave
after drying (Na.sub.2SO.sub.4) and concentration in vacuo the
desired product which was subjected to flash chromatography (20%
EtOAc/80% hexanes) to provide 480 mg (100%) of pure dipeptide.
.sup.1H NMR (CDCl.sub.3) .delta. 4.2 (d, 2H); 4.0 (t, 1H); 3.5 (m,
1H); 2.0 (m, 3H0; 2.8 (m, 2H); 1.5 (s, 9H); 1.1 (s, 9H); 1.0 (d,
3H); 0.9 (d, 3H) ppm.
[0500] The Cbz group of the dipeptide was removed as described
above and the resulting aminoester dipeptide was coupled to
Cbz-cyclohexyl glycine shown in the next step.
[0501] To a solution of Cbz-cyclohexyl glycine (289 mg, 1 mmol) in
2 mL of DCM at 0.degree. C. was added EDC (228 mg, 1.19 mmol), HOBt
(190 mg, 1.29 mmol) and DIEA (0.517 mL, 2.97 mmol). The resulting
mixture was stirred at 0.degree. C. for 15 min. after which, the
above amino ester was slowly added in 2 mL of DCM. The resulting
reaction mixture was stirred at rt for 16 h. Concentrated to a
residue that was redissolved in EtOAc. Successive washes with 0.5N
HCL, satd' aqueous NaHCO.sub.3 and brine gave after drying
(Na.sub.2SO.sub.4) and concentration in vacuo the desired product
which was subjected to flash chromatography (20% EtOAc/80% hexanes)
to provide 556 mg (90%) of pure tripeptide. The Cbz group of the
tripeptide was removed as described above and the resulting
aminoester tripeptide was coupled to 1,4-pyrazine carboxylic acid
shown in the next step.
[0502] To a solution of 1,4-pyrazine carboxylic acid (110 mg, 0.891
mmol)) in 2 mL of DCM was added PyBrOP (457 mg, 0.98 mmol and DIEA
(0.465 mL, 2.67 mmol). The resulting mixture was stirred at rt for
15 min. after which, the above amino ester was slowly added in 2 mL
of DCM. The resulting reaction mixture was stirred at rt for 16 h.
Concentrated to a residue that was redissolved in EtOAc. Successive
washes with 0.5N HCL, sat'd aqueous NaHCO.sub.3 and brine gave
after drying (Na.sub.2SO.sub.4) and concentration in vacuo the
desired product which was subjected to flash chromatography (50%
EtOAc/50% hexanes) to provide 410 mg (79%) of pure capped
tripeptide with consistent .sup.1H NMR (CDCl.sub.3).
[0503] The t-butyl ester group of the capped tripeptide (410 mg,
0.688 mmol) was cleaved with a 1:1 mixture of TFA-DCM at rt for 45
minutes and concentrated in vacuo. The resulting aminoester
tripeptide was coupled to hydroxyamide detailed in the next
step.
[0504] To a stirring solution of the capped tripeptide acid from
above in 6 mL of dry DMF at 0.degree. C. was added, PyBOP (376 mg,
0.722 mmol) followed by NMM (0.226 mL, 2.06 mmol). The reaction
mixture was stirred for 1 h at rt after which a solution of
hydroxyamide (168 mg, 0.758 mmol) and 0.226 mL of NMM was slowly
added. The coupling reaction was stirred for 16 h, diluted with
ethyl acetate and was successively washed with; water (3.times.),
citric acid 10%, water and brine. The organic layer was dried
(Na.sub.2SO.sub.4) and concentrated in vacuo. Flash chromatography
(2.5% MeOH/97.5% ethyl acetate) provided 362 mg of hydroxy amide
tetrapeptide that was oxidized with Dess-Martin periodinane reagent
(650 mg, 1.53 mmol) and t-butanol (0.65 mL) in 5 mL of DCM at rt
for 3 h. The reaction mixture was quenched with sodium thiosulfate
1M solution (2 mL) and stirred until the two phases are clearly
separated. The organic layer was diluted with 5 more mL of DCM and
washed (3.times.) with 10% potassium carbonate aqueous solution (5
mL), dried (Na.sub.2SO.sub.4) and concentrated in vacuo. Flash
chromatography (2.5% MeOH/97.5% ethyl acetate) provided 270 mg of
ketoamide tetrapeptide 56. LCMS M+H=706.42, M-H=704.42. Retention
Time (10-90% MeCN--H.sub.2O with 0.1% TFA over 9 minutes)=7.73-8.81
min. LCMS M+H=682.2.
Example 2
3-tert-Butyl-2-(tert-butyl-dimethyl-silanyloxymethyl)-5-oxo-pyrrolidine-1--
carboxylic acid tert-butyl ester (6a)
[0505] t-Butyl zinc bromide 0.5M solution in THF (3.7 mL, 1.83
mmol)was added to a solution of enone 4a (280 mg, 0.85 mmol) in THF
containing BF.sub.3OEt.sub.2 (350 uL, 2.75 mmol) and TMSCl (465 uL)
at -30.degree. C. over a period of 5 minutes. The heterogeneous
mixture was stirred at -30.degree. C. for 3.5 h than quenched with
sat'd NH.sub.4Cl solution. Extracted with ether (3.times.) and the
combined extract were washed with brine, dried with sodium sulfate
and concentrated in vacuo. Flash chromatography (10% ethyl
acetate-hexanes) provided 210 mg (64%) of 6a. .sup.1H NMR
(CDCl.sub.3) .delta. 3.9 (s, 1H); 3.8 (dd, 1H); 3.5 (d, 1H); 2.8
(dd, 1H); 2.3 (d, 1H); 1.9 (d, 1H); 1.4 (s, 9H); 0.9 (s, 18H); 0.1
(s, 3H); 0.05 (s, 3H) ppm.
Example 3
3-Benzyl-2-(tert-butyl-dimethyl-silanyloxymethyl)-5-oxo-pyrrolidine-1-carb-
oxylic acid tert-butyl ester (7a)
[0506] To a mixture of n-butyllithium (5.5 mL, 0.0086 mol) and THF
at -78.degree. C., was added TMEDA and benzylphenylsulfide (1.91 g,
0.0095 mol). The colorless solution turned pale yellow. After 15
min of stirring at -78.degree. C., the pyrolidone 4 (2.4 g, 0.0073
mol) in 10 mL of THF was added dropwise. After the addition was
completed, the reaction mixture was stirred for 1 h at -78.degree.
C. The reaction was quenched with satd' NH.sub.4Cl solution and the
mixture warmed to rt and poured into water. Ether mixture was
extracted with ethyl ether and the organic phase was washed with
brine, dried and concentrated in vacuo. Flash chromatography (20%
ethyl acetate-hexane) provided 1.69 g (45%) of the desired
intermediate. Reduction with 16.9 g of Ra--Ni in refluxing
acetone-water (1:1) for 12 h provided, after chromatography (2%
acetone-chloroform), 1.11 g (83%) of the desired compound 7.
.sup.1H NMR (CDCl.sub.3) .delta. 7.3 (m, 5H); 3.8 (m, 2H); 3.7 (d,
1H); 2.7-2.9 (m, 3H); 2.1 (m, 2H); 1.5 (s, 9H); 1.7 (s, 9H); 0.1
(s, 6H) ppm.
Example 4
Pyrazine-2-carboxylic acid
({1-[3-benzyl-2-(1-cyclopropylaminooxalyl-butylcarbamoyl)-pyrrolidine-1-c-
arbonyl]-2,2-dimethyl-propylcarbamoyl}-cyclohexyl-methyl)-amide
(65)
[0507] Prepared as described above in scheme 1 starting with
intermediate 7a to give 65 with consistent analytical data.
Retention Time (10-90% MeCN--H.sub.2O with 0.1% TFA over 6
minutes)=8.0-9.2 min. LCMS M+H=730.2
Example 5
3-Cyclohexyl-pyrrolidine-2-carboxylic acid tert-butyl ester
(12a)
[0508] 3-Phenyl proline 10a was hydrogenated with catalytic
platinum oxide in ethanol/acetic acid/water (7/2/1) under 50 psi of
hydrogen for 18 h to give 3-cyclohexyl proline quantitatively.
Compound 12a was prepared according to benzoylation and
esterification from example 1; step 3.
Example 6
3-Cyclopropylmethyl-pyrrolidine-1,2-dicarboxylic acid 1-tert-butyl
ester 2-methyl ester (A)
[0509] In a round bottom flask, a solution of the allyl proline
(358 mg, 1.33 mmol) in dry DCE was cooled to 0.degree. C., and
diethyl zinc in hexanes 15% (5.5 mL, 6.63 mmol) was added slowly
via a syringe. To this solution was added chloroiodomethane (967
uL, 13.3 mmol) dropwise. The solution was stirred at 0.degree. C.
for 20 minutes, allowed to warm to rt an stirred for 1 h. The
reaction mixture was cooled to 0.degree. C. and quenched with satd'
NH.sub.4Cl solution and stirred vigorously for 10 minutes.
Extracted with dichloromethane, dried with sodium sulfate and
concentrated in vacuo. Chromatography (20% ethyl acetate-hexanes)
gave 65 mg (17%) of the desired product A. .sup.1H NMR (CDCl.sub.3)
.delta. 3.8 (s, 1H); 3.7 (s, 3H); 3.6-3.4 (m, 2H); 2.4 (m, 1H); 2.3
(m, 1H); 1.3 (m, 3H); 0.8 (m, 1H); 0.5 (m, 2H); 0.2 (m, 2H)
ppm.
Example 7
3-Cyclopropylmethyl-1-(3-methyl-2-{3-methyl-2-[(pyrazine-2-carbonyl)-amino-
]-butyrylamino}-butyryl)-pyrrolidine-2-carboxylic acid methyl ester
(C)
[0510] Tripeptide C was prepared by the coupling of
cyclopropylmethyl proline A (41 mg, 0.16 mmol) and capped dipeptide
B (52 mg, 0.16 mmol) with EDC/HOAt to give 60 mg (77%) of the
desired tripeptide C after chromatography (1:1 ethyl
acetate-hexanes). .sup.1H NMR (CDCl.sub.3) .delta. 9.4 (s, 1H); 8.8
(s, 1H); 8.5 (s, 1H); 8.2 (d, 1H); 6.5 (d, 1H); 4.6 (t, 1H); 4.5
(t, 1H); 4.2 (m, 1H); 3.8 (s, 3H); 3.7 (m, 2H); 2.3 (m, 4H); 2.2
(m, 2H); 1.5 (s, 12H); 1.3 (m, 2H); 1.0 (m, 2H); 0.5 (m, 2H)
ppm.
Example 8
2-(3-{[3-Cyclopropylmethyl-1-(3-methyl-2-{3-methyl-2-[(pyrazine-2-carbonyl-
)-amino]-butyrylamino}-butyryl)-pyrrolidine-2-carbonyl]-amino}-2-oxo-hexan-
oylamino)-3-phenyl-propionic acid (D)
[0511] Prepared as in example 1. Retention Time (10-90%
MeCN--H.sub.2O with 0.1% TFA over 6 minutes)=7.55-7.78 min. LCMS
M+H=748.3.
Example 9
HCV Replicon Cell Assay Protocol
[0512] Cells containing hepatitis C virus (HCV) replicon were
maintained in DMEM containing 10% fetal bovine serum (FBS), 0.25 mg
per ml of G418, with appropriate supplements (media A).
[0513] On day 1, replicon cell monolayer was treated with a
trypsin:EDTA mixture, removed, and then media A was diluted into a
final concentration of 100,000 cells per ml with. 10,000 cells in
100 ul were plated into each well of a 96-well tissue culture
plate, and cultured overnight in a tissue culture incubator at
37.degree. C.
[0514] On day 2, compounds (in 100% DMSO) were serially diluted
into DMEM containing 2% FBS, 0.5% DMSO, with appropriate
supplements (media B). The final concentration of DMSO was
maintained at 0.5% throughout the dilution series.
[0515] Media on the replicon cell monolayer was removed, and then
media B containing various concentrations of compounds was added.
Media B without any compound was added to other wells as no
compound controls.
[0516] Cells were incubated with compound or 0.5% DMSO in media B
for 48 hours in a tissue culture incubator at 37.degree. C. At the
end of the 48-hour incubation, the media was removed, and the
replicon cell monolayer was washed once with PBS and stored at
-80.degree. C. prior to RNA extraction.
[0517] Culture plates with treated replicon cell monolayers were
thawed, and a fixed amount of another RNA virus, such as Bovine
Viral Diarrhea Virus (BVDV) was added to cells in each well. RNA
extraction reagents (such as reagents from RNeasy kits) were added
to the cells immediately to avoid degradation of RNA. Total RNA was
extracted according the instruction of manufacturer with
modification to improve extraction efficiency and consistency.
Finally, total cellular RNA, including HCV replicon RNA, was eluted
and stored at -80.degree. C. until further processing.
[0518] A Taqman real-time RT-PCR quantification assay was set up
with two sets of specific primers and probe. One was for HCV and
the other was for BVDV. Total RNA extractants from treated HCV
replicon cells was added to the PCR reactions for quantification of
both HCV and BVDV RNA in the same PCR well. Experimental failure
was flagged and rejected based on the level of BVDV RNA in each
well. The level of HCV RNA in each well was calculated according to
a standard curve run in the same PCR plate. The percentage of
inhibition or decrease of HCV RNA level due to compound treatment
was calculated using the DMSO or no compound control as 0% of
inhibition. The IC.sub.50 (concentration at which 50% inhibition of
HCV RNA level is observed) was calculated from the titration curve
of any given compound.
Example 10
HCV Ki Assay Protocol
[0519] HPLC Microbore method for separation of 5AB substrate and
products
Substrate:
NH.sub.2-Glu-Asp-Val-Val-(alpha)Abu-Cys-Ser-Met-Ser-Tyr-COOH
[0520] A stock solution of 20 mM 5AB (or concentration of your
choice) was made in DMSO w/0.2M DTT. This was stored in aliquots at
-20 C.
[0521] Buffer: 50 mM HEPES, pH 7.8; 20% glycerol; 100 mM NaCl
[0522] Total assay volume was 100 .mu.L
TABLE-US-00001 X1 conc. in (.mu.L) assay Buffer 86.5 see above 5 mM
KK4A 0.5 25 .mu.M 1 M DTT 0.5 5 mM DMSO or inhibitor 2.5 2.5% v/v
50 .mu.M tNS3 0.05 25 nM 250 .mu.M 5AB 20 25 .mu.M (initiate)
[0523] The buffer, KK4A, DTT, and tNS3 were combined; distributed
78 .mu.L each into wells of 96 well plate. This was incubated at 30
C for .about.5-10 min.
[0524] 2.5 .mu.L of appropriate concentration of test compound was
dissolved in DMSO (DMSO only for control) and added to each well.
This was incubated at room temperature for 15 min.
[0525] Initiated reaction by addition of 20 .mu.L of 250 .mu.M 5AB
substrate (25 .mu.M concentration is equivalent or slightly lower
than the Km for 5AB).
[0526] Incubated for 20 min at 30 C.
[0527] Terminated reaction by addition of 25 .mu.L of 10% TFA
[0528] Transferred 120 .mu.L aliquots to HPLC vials
[0529] Separated SMSY product from substrate and KK4A by the
following method:
Microbore Separation Method:
Instrumentation: Agilent 1100
Degasser G1322A
[0530] Binary pump G1312A
Autosampler G1313A
[0531] Column thermostated chamber G1316A Diode array detector
G1315A
Column:
[0532] Phenomenex Jupiter; 5 micron C18; 300 angstroms; 150.times.2
mm; P/O 00F-4053-B0 Column thermostat: 40 C Injection volume: 100
.mu.L Solvent A=HPLC grade water+0.1% TFA Solvent B=HPLC grade
acetonitrile+0.1% TFA
TABLE-US-00002 Time Flow Max (min) % B (ml/min) press. 0 5 0.2 400
12 60 0.2 400 13 100 0.2 400 16 100 0.2 400 17 5 0.2 400
Stop time: 17 min Post-run time: 10 min.
[0533] Table 1 below depicts IC.sub.50 data for certain compounds
of the invention.
Compounds with Ki's ranging from 0.5 .mu.M to >1 .mu.M are
designated A. Compounds with Ki's ranging from 0.5 .mu.M to 0.1
.mu.M are designated B. Compounds with Ki's below 0.1 .mu.M are
designated C. Compounds with IC.sub.50's ranging from 1 .mu.M to
>10 .mu.M are designated A. Compounds with IC.sub.50's ranging
from 1 .mu.M to 0.5 .mu.M are designated B. Compounds with
IC.sub.50's below 0.5 .mu.M are designated C. ND means no data.
TABLE-US-00003 TABLE 1 Compound Ki IC.sub.50 5 C ND 15 B ND 16 B ND
19 B ND 20 B ND 22 B ND 25 B ND 26 C ND 27 C ND 28 C ND 29 C ND 30
B ND 31 C ND 32 B ND 33 B ND 35 A ND 36 B ND 39 C B 41 A C 42 C ND
43 B ND 44 B B 45 A ND 46 B ND 50 B ND 51 B ND 52 C C 53 B ND 54 B
A 55 C B 56 B ND 57 C ND 60 ND A 62 C B 63 C B 64 B A 65 C B 66 B B
67 A A 68 C C 69 C C 71 C A 72 C B 73 B A 74 B A 75 B A 76 B A 77 B
A
* * * * *