U.S. patent application number 10/094790 was filed with the patent office on 2003-04-10 for novel sulfonamide inhibitors of aspartyl protease.
This patent application is currently assigned to Vertex Pharmaceuticals, Inc.. Invention is credited to Bhisetti, Govinda R., Murcko, Mark A., Tung, Roger D..
Application Number | 20030069222 10/094790 |
Document ID | / |
Family ID | 25477403 |
Filed Date | 2003-04-10 |
United States Patent
Application |
20030069222 |
Kind Code |
A1 |
Tung, Roger D. ; et
al. |
April 10, 2003 |
Novel sulfonamide inhibitors of aspartyl protease
Abstract
The present invention relates to a novel class of sulfonamides
which are aspartyl protease inhibitors. In one embodiment, this
invention relates to a novel class of HIV aspartyl protease
inhibitors characterized by specific structural and physicochemical
features. This invention also relates to pharmaceutical
compositions comprising these compounds. The compounds and
pharmaceutical compositions of this invention are particularly well
suited for inhibiting HIV-1 and HIV-2 protease activity and
consequently, may be advantageously used as anti-viral agents
against the HIV-1 and HIV-2 viruses. This invention also relates to
methods for inhibiting the activity of HIV aspartyl protease using
the compounds of this invention and methods for screening compounds
for anti-HIV activity.
Inventors: |
Tung, Roger D.; (Arlington,
MA) ; Murcko, Mark A.; (Holliston, MA) ;
Bhisetti, Govinda R.; (Lexington, MA) |
Correspondence
Address: |
FISH & NEAVE
1251 AVENUE OF THE AMERICAS
50TH FLOOR
NEW YORK
NY
10020-1105
US
|
Assignee: |
Vertex Pharmaceuticals,
Inc.
|
Family ID: |
25477403 |
Appl. No.: |
10/094790 |
Filed: |
March 8, 2002 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10094790 |
Mar 8, 2002 |
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08484326 |
Jun 7, 1995 |
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6372778 |
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08484326 |
Jun 7, 1995 |
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08142327 |
Nov 24, 1993 |
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5585397 |
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08142327 |
Nov 24, 1993 |
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07941982 |
Sep 8, 1992 |
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Current U.S.
Class: |
514/211.01 ;
514/217.12; 514/227.5; 514/231.2; 514/252.12; 514/331; 514/365;
514/374; 514/385 |
Current CPC
Class: |
C07C 311/13 20130101;
C07D 263/24 20130101; C07D 413/12 20130101; C07C 311/39 20130101;
C07K 5/06139 20130101; C07D 271/08 20130101; A61K 38/00 20130101;
C07C 311/29 20130101; C07D 403/12 20130101; C07D 413/14 20130101;
C07D 213/78 20130101; C07D 409/14 20130101; C07D 207/26 20130101;
C07D 231/14 20130101; C07D 307/12 20130101; C07C 2601/02 20170501;
C07C 2601/08 20170501; C07D 213/30 20130101; C07D 307/20 20130101;
C07D 215/38 20130101; C07D 405/12 20130101; C07C 307/06 20130101;
C07D 409/12 20130101; C07D 417/12 20130101; C07D 215/48 20130101;
C07D 239/22 20130101; C07C 311/18 20130101; A61P 31/18 20180101;
C07C 311/46 20130101; C07C 2601/14 20170501; C07D 277/36 20130101;
A61P 31/12 20180101; C07D 209/08 20130101 |
Class at
Publication: |
514/211.01 ;
514/217.12; 514/227.5; 514/231.2; 514/252.12; 514/331; 514/365;
514/374; 514/385 |
International
Class: |
A61K 031/553; A61K
031/554; A61K 031/55; A61K 031/54; A61K 031/535; A61K 031/445 |
Claims
We claim:
1. A compound of formula I: 610wherein: A is selected from the
group consisting of H; Het; -R.sup.1-Het;
--R.sup.1--C.sub.1-C.sub.6 alkyl, which may be optionally
substituted with one or more groups selected from the group
consisting of hydroxy, C.sub.1-C.sub.4 alkoxy, Het, --O-Het,
--NR.sup.2--CO--N(R.sup.2)(R.sup.2) and --CO--N(R.sup.2)(R.sup.2);
and --R.sup.1--C.sub.2-C.sub.6 alkenyl, which may be optionally
substituted with one or more groups selected from the group
consisting of hydroxy, C.sub.1-C.sub.4 alkoxy, Het, --O-Het,
--NR.sup.2--CO--N(R.sup.2)(R.sup.2) and --CO--N(R.sup.2)(R.sup.2);
each R.sup.1 is independently selected from the group consisting of
--C(O)--, --S(O).sub.2--, --C(O)--C(O)--, --O--C(O)--,
--O--S(O).sub.2, --NR.sup.2--S(O).sub.2--, --NR.sup.2--C(O)-- and
--NR.sup.2--C(O)--C(O)--; each Het is independently selected from
the group consisting of C.sub.3-C.sub.7 cycloalkyl; C.sub.5-C.sub.7
cycloalkenyl; C.sub.6-C.sub.10 aryl; and 5-7 membered saturated or
unsaturated heterocycle, containing one or more heteroatoms
selected from N, N(R.sup.2), O, S and S(O).sub.n, wherein said
heterocycle may optionally be benzofused; and wherein any member of
said Het may be optionally substituted with one or more
substituents selected from the group consisting of oxo, --OR.sup.2,
--R.sup.2, --N(R.sup.2)(R.sup.2), --R.sup.2--OH, --CN,
--CO.sub.2R.sup.2, --C(O)--N(R.sup.2)(R.sup.2),
--S(O).sub.2--N(R.sup.2)(R.sup.2), --N(R.sup.2)--C(O)--R.sub.2,
--C(O)--R.sup.2, --S(O).sub.n--R.sup.2, --OCF.sub.3,
--S(O).sub.n--Ar, methylenedioxy, --N(R.sup.2)--S(O).sub.2(R-
.sup.2), halo, --CF.sub.3, --NO.sub.2, Ar and --O--Ar; each R.sup.2
is independently selected from the group consisting of H and
C.sub.1-C.sub.3 alkyl optionally substituted with Ar; B, when
present, is --N(R.sup.2)--C(R.sup.3) (R.sup.3)--C(O)--; x is 0 or
1; each R.sup.3 is independently selected from the group consisting
of H, Het, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.3-C.sub.6 cycloalkyl and C.sub.5-C.sub.6 cycloalkenyl,
wherein any member of said R.sup.3, except H, may be optionally
substituted with one or more substituents selected from the group
consisting of --OR.sup.2, --C(O)--NH--R.sup.2,
--S(O).sub.n--N(R.sup.2)(R.sup.2), Het, --CN, --SR.sup.2,
--CO.sub.2R.sup.2, NR.sup.2--C(O)--R.sup.2; each n is independently
1 or 2; D and D' are independently selected from the group
consisting of Ar; C.sub.1-C.sub.4 alkyl, which may be optionally
substituted with one or more groups selected from C.sub.3-C.sub.6
cycloalkyl, --OR.sub.2, --R.sup.3, --O--Ar and Ar; C.sub.2-C.sub.4
alkenyl, which may be optionally substituted with one or more
groups selected from the group consisting of C.sub.3-C.sub.6
cycloalkyl, --OR.sup.2, --R.sup.3, --O--Ar and Ar; C.sub.3-C.sub.6
cycloalkyl, which may be optionally substituted with or fused with
Ar; and C.sub.5-C.sub.6 cycloalkenyl, which may be optionally
substituted with or fused with Ar; each Ar is independently
selected from the group consisting of phenyl; 3-6 membered
carbocyclic ring and 5-6 membered heterocyclic ring containing one
or more heteroatoms selected from O, N, S, S(O).sub.n and
N(R.sup.2), wherein said carbocyclic or heterocyclic ring may be
saturated or unsaturated and optionally substituted with one or
more groups selected from the group consisting of oxo, --OR.sup.2,
--R.sup.2, --N(R.sup.2)(R.sup.2), --N(R.sup.2)--C(O)--R.sup.2,
--R.sup.2--OH, --CN, --CO.sub.2R.sup.2,
--C(O)--N(R.sup.2)(R.sup.2), halo and --CF.sub.3; E is selected
from the group consisting of Het; O-Het; Het-Het; --O--R.sup.3;
--NR.sup.2R.sup.3; C.sub.1-C.sub.6 alkyl, which may be optionally
substituted with one or more groups selected from the group
consisting of R.sup.4 and Het; C.sub.2-C.sub.6 alkenyl, which may
be optionally substituted with one or more groups selected from the
group consisting of R.sup.4 and Het; C.sub.3-C.sub.6 saturated
carbocycle, which may optionally be substituted with one or more
groups selected from the group consisting of R.sup.4 and Het; and
C.sub.5-C.sub.6 unsaturated carbocycle, which may optionally be
substituted with one or more groups selected from the group
consisting of R.sup.4 and Het; and each R.sup.4 is independently
selected from the group consisting of , --C(O)--NHR.sup.2,
--S(O).sub.2--NHR.sup.2, halo, --NR.sup.2--C(O)--R.sup- .2 and
--CN.
2. The compound according to claim 1, wherein said compound has the
structure of formula XXII: 611wherein A, D' and E are defined as in
claim 1.
3. The compound according to claim 1, wherein said compound has the
structure of formula XXIII: 612wherein x, Het, R.sup.3, D' and E
are defined as in claim 1.
4. The compound according to claim 1, wherein said compound has the
structure of formula XXXI: 613wherein A, R.sup.3, D' and E are
defined as in claim 1.
5. A compound of formula I, wherein: A is selected from the group
consisting of H; --R.sup.1-Het; --R.sup.1--C.sub.1-C.sub.6 alkyl,
which may be optionally substituted with one or more groups
selected from the group consisting of hydroxy, C.sub.1-C.sub.4
alkoxy, Het and --O-Het; and --R.sup.1--C.sub.2-C.sub.6 alkenyl,
which may be optionally substituted with one or more groups
selected from hydroxy, C.sub.1-C.sub.4 alkoxy, Het and --O-Het;
each R.sup.1 is independently selected from the group consisting of
--C(O)--, --S(O).sub.2--, --C(O)--C(O)--, --O--CO--,
--O--S(O).sub.2-- and --NR.sup.2--S(O).sub.2--; each Het is
independently selected from the group consisting of C.sub.3-C.sub.7
cycloalkyl; C.sub.5-C.sub.7 cycloalkenyl; C.sub.6-C.sub.10 aryl;
and 5-7 membered saturated or unsaturated heterocycle, containing
one or more heteroatoms selected from N, O and S, which may
optionally be benzofused; wherein any member of said Het may be
optionally substituted with one or more substituents selected from
the group consisting of oxo, --OR.sup.2, R.sup.2,
--N(R.sup.2).sub.2, --R.sup.2--OH, --CN, --CO.sub.2R.sup.2.sub.1
--C(O)--N(R.sup.2).sub.2 and --S(O).sub.2--N(R.sup.2).sub.2; each
R.sup.2 is independently selected from the group consisting of H
and C.sub.1-C.sub.3 alkyl; B, when present, is
--NH--CH(R.sup.3)--C(O)--; x is 0 or 1; R.sup.3 is selected from
the group consisting of Het, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.3-C.sub.6 cycloalkyl and C.sub.5-C.sub.6
cycloalkenyl, wherein any member of said R.sup.3 may be optionally
substituted with one or more substituents selected from the group
consisting of --OR.sup.2, --C(O)--NH--R.sup.2,
--S(O).sub.n--N(R.sup.2).sub.2, Het and --CN; n is 1 or 2; D and D'
are independently selected from the group consisting of Ar;
C.sub.1-C.sub.4 alkyl, which may be optionally substituted with
C.sub.3-C.sub.6 cycloalkyl or Ar; C.sub.2-C.sub.4 alkenyl, which
may be optionally substituted with C.sub.3-C.sub.6 cycloalkyl or
Ar; C.sub.3-C.sub.6 cycloalkyl, which may be optionally substituted
or fused with Ar; and C.sub.1-C.sub.6 cycloalkenyl, which may be
optionally substituted or fused with Ar; with the proviso that when
D is attached to N, D may not be methyl or C.sub.2 alkenyl; Ar is
selected from the group consisting of phenyl; 3-6 membered
carbocyclic ring and 5-6 membered heterocyclic ring containing one
or more heteroatoms selected from O, N and S, wherein said
carbocyclic or heterocyclic ring may be saturated or unsaturated
and optionally substituted with one or more groups selected from
the group consisting of oxo, --OR.sup.2, --R.sup.2,
--N(R.sup.2).sub.2, --N(R.sup.2)--C(O)R.sup.2, --R.sup.2--OH, --CN,
--CO.sub.2R.sup.2, --C(O)--N(R.sup.2).sub.2, halo and --CF.sub.3; E
is selected from the group consisting of Het; --O--R.sup.3;
--NR.sup.2R.sup.5; C.sub.1-C.sub.6 alkyl, which may be optionally
substituted with one or more R.sup.4 or Het; C.sub.2-C.sub.6
alkenyl, which may be optionally substituted with one or more
R.sup.4 or Het; C.sub.3-C.sub.6 saturated carbocycle, which may
optionally be substituted with one or more R.sup.4 or Het; and
C.sub.5-C.sub.6 unsaturated carbocycle, which may optionally be
substituted with one or more R.sup.4 or Het; each R.sup.4 is
independently selected from the group consisting of ,
--C(O)--NHR.sup.2, --S(O).sub.2--NHR.sup.2, halo and --CN; and each
R is independently selected from the group consisting of H and
R.sup.3, with the proviso that at least one R.sup.5 is not H.
6. The compound according to claim 2, wherein: A is R.sup.1-Het;
and D' is selected from the group consisting of C.sub.1-C.sub.3
alkyl and C.sub.3 alkenyl, wherein said alkyl or alkenyl may
optionally be substituted with one or more groups selected from the
group consisting of C.sub.3-C.sub.6 cycloalkyl, --OR.sup.2, --O--Ar
and Ar.
7. The compound according to claim 3, wherein: R.sup.3 is selected
from the group consisting of C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.5-C.sub.6 cycloalkyl, C.sub.5-C.sub.6 cycloalkenyl
and a 5-6 membered saturated or unsaturated heterocycle, wherein
any member of said R.sup.3 may optionally be substituted with one
or more substituents selected from the group consisting of
--OR.sup.2, --C(O)--NH--R.sup.2,
--S(O).sub.nN(R.sup.2)(R.sup.2).sub.2, Het, --CN, --SR.sup.2,
--C(O).sub.2R.sup.2, NR.sup.2--C(O)--R.sup.2; and D' is selected
from the group consisting of C.sub.1-C.sub.3 alkyl and C.sub.3
alkenyl, wherein said alkyl or alkenyl may optionally be
substituted with one or more groups selected from the group
consisting of C.sub.3-C.sub.6 cycloalkyl, --OR.sup.2, --O--Ar and
Ar.
8. The compound according to claim 4, wherein: A is R.sup.1-Het;
each R.sup.3 is independently C.sub.1-C.sub.6 alkyl, which may be
optionally substituted with a substituent selected from the group
consisting of --OR.sup.2.sub.1 --C(O)--NH--R.sup.2,
--S(O).sub.nN(R.sup.2).sub.2, Het, --CN, --SR.sup.2,
--CO.sub.2R.sup.2. --NR.sup.2--C(O)--R.sup.2; and D' is
C.sub.1-C.sub.4 alkyl, which may be optionally substituted with a
group selected from the group consisting of C.sub.3-C.sub.6
cycloalkyl, --OR.sup.2, --O--Ar and Ar; and E is selected from the
group consisting of Het, Het-Het and --NR.sup.2R.sup.3.
9. A compound selected from the group consisting of:
(S)--N-1-(3-((3-Acetylamino-4-fluorobenzenesulfonyl)-benzyl-amino)-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinam-
ide and
(S)--N-1-(3-((4-Acetylamino-3-fluorobenzenesulfonyl)-benzyl-amino)-
-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-su-
ccinamide (compounds 2);
(S)--N-1-(3-((5-Acetylamino-3-methyl-thiophene-2--
sulfonyl)-benzyl-amino)-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinolin-
e-2-carbonyl)-amino)-succinamide (compound 5);
(S)--N-1-(1-Benzyl-3-(benzy-
l-(5-isoxazol-3-yl-thiophene-2-sulfonyl)-amino)-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinam-
ide (compound 6);
(S)--N-1-(3-((Benzo(1,2,5)oxadiazole-4-sulfonyl)-benzyl--
amino)-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-ami-
no)-succinamide (compound 9);
N-1-(1-(S)-Benzyl-3-(benzyl-(3-sulfamoyl-ben-
zenesulfonyl)-amino)-2-(syn)-hydroxy-propyl)-2-((quinoline-2-carbonyl)-ami-
no)-succinamide (compound 10);
(S)--N-1-(1-(S)-Benzyl-2-(syn)-hydroxyl-3-(-
isobutyl-(5-pyridin-2-yl-thiophene-2-sulfonyl)-amino)-propyl)-2-((quinolin-
e-2-carbonyl)-amino)-succinamide (compound 12);
(S)--N-1-(3-((4-Benzenesul-
fonyl-thiophene-2-sulfonyl)-isobutyl-amino)-(1S,2
syn)-1-benzyl-2-hydroxy--
propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide (compound 13);
(S)--N-1-(1-(S)-Benzyl-3-((4-fluoro-benzenesulfonyl)-isobutyl-amino)-2-(s-
yn)-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide
(compound 14);
(S)--N-1-(3-((4-Acetylamino-3-fluorobenzenesulfonyl)-isobutyl-amino)-
-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-su-
ccinamide (compound 15);
(S)--N-1-(3-((3-Acetylamino-4-fluorobenzenesulfon-
yl)-isobutyl-amino)-(1S,2 syn)-1-benzyl
2-hydroxy-propyl)-2-((quinoline-2-- carbonyl)-amino)-succinamide
(compound 16); (S)--N-1-(1-(S)-Benzyl-3-((4-a-
cetylamino-benzenesulfonyl)-isobutyl-amino)-2-(syn)-hydroxy-propyl)-2-((qu-
inoline-2-carbonyl)-amino)-succinamide (compound 17);
(S)--N-1-(3-((5-Acetylamino-3-methyl-thiophene-2-sulfonyl)-isobutyl-amino-
)-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-s-
uccinamide (compound 18);
(S)--N-1-(3-((3-Acetylamino-benzenesulfonyl)-iso-
butyl-amino)-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbony-
l)-amino)-succinamide (compound 19);
(S)--N-1-(3-((Benzo(1,2,5)oxadiazole--
4-sulfonyl)-isobutyl-amino)-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quin-
oline-2-carbonyl)-amino)-succinamide (compound 20); N-1-((1S-2
syn)-1-Benzyl-2-hydroxy-3-(1-isobutyl-3,3-dimethylsulfonylurea)-propyl)-2-
-((quinoline-2-carbonyl)-amino)-succinamide (compound 21);
N-1-(3-((4-Acetylamino-benzenesulfonyl)-isobutyl-amino)-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-(pyridin-2-yl-methoxycarbonyl)-succinam-
ide (compound 22);
N-1-(3-((4-Acetylamino-benzenesulfonyl)-isobutyl-amino)- -(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-(pyridin-4-yl-methoxycarbonyl)-su-
ccinamide (compound 23);
N-1-(3-((4-Fluoro-benzenesulfonyl)-isobutyl-amino- )-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-(pyridin-2-yl-methoxycarbonyl)-s-
uccinamide (compound 26); 4-Fluoro-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S-
)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonami-
de (compound 35); 3,4-Dichloro-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-te-
trahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide
(compound 37); N-(4-(((2 syn,
3S)-2-Hydroxy-4-phenyl-3-(pyridin-3-yl-meth-
oxycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide
(compound 44); 2,4-Dimethyl-thiazole-5-sulfonic
acid-(1,1-dimethyl-ethoxycarbonylam- ino)-(2 syn,
3S)-2-hydroxy-4-phenyl-butyl)-isobutyl-amide (compound 46);
N-(4-(((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbo-
nylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide (compound
48); 4-Fluoro-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((R)-tetrahydrofuran-3-yloxy-
carbonylamino)-butyl)-N-isobutyl-benzenesulfonamide and
4-Fluoro-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((R)-tetrahydrofuran-3-yloxycarbonylamino)--
butyl)-N-isobutyl-benzenesulfonamide (compounds 52); Benzo
(1,2,5)oxadiazole-5-sulfonic acid ((2 syn,
3S)-2-hydroxy-4-phenyl-3-(pyri-
din-3-yl-methoxycarbonylamino)-butyl)-isobutylamide (compound 66);
N-(4-(((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((R)-tetrahydrofuran-3-yloxycarbo-
nylamino)-butyl)-isobutyl-sulfamoyl-phenyl)-acetamide and N-(4-(((2
syn,
3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl-
)-isobutyl-sulfamoyl)-phenyl)-acetamide (compounds 86);
N-(2-Fluoro-5-(((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)--
butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide (compound 88);
N-(3-(((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl-
)-isobutyl-sulfamoyl)-phenyl)-acetamide (compound 91);
4-Fluoro-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((R)-tetrahydrofuran-3-yloxycarbonylamino)-butyl-
)-N-isobutyl-benzenesulfonamide (compound 93);
N-(4-(((syn)-2-Hydroxy-(S)--
4-phenyl-3-((tetrahydro-furan-(R)-3-yl)-oxycarbonylamino)-butyl)-isobutyl--
sulfamoyl)-phenyl)-acetamide (compound 94); 4-Fluoro-N-(2 syn,
3S)-2-hydroxy-4-phenyl-3-((tetrahydro-furan-(R)-3-ylmethoxycarbonylamino)-
-butyl)-N-isobutyl-benzenesulfonamide and 4-Fluoro-N-(2 syn,
3S)-2-hydroxy-4-phenyl-3-((tetrahydro-furan-(S)-3-ylmethoxycarbonylamino)-
-butyl)-N-isobutyl-benzenesulfonamide (compounds 97);
4-Fluoro-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-(pyridin-3-yl-methoxycarbonylamino)-butyl)-N-iso-
butyl-benzenesulfonamide (compound 98); 4-Chloro-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl-
)-isobutyl-benzenesulfonamide (compound 99); N-((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl-
)-N-isobutyl-4-methoxy-benzenesulfonamide (compound 100);
4-Fluoro-N-(2-(syn)-hydroxy-3-((2-oxazolidon-(S)-4-yl)-methoxycarbonylami-
no)-4-(S)-phenyl-butyl)-N-isobutyl-benzenesulfonamide (compound
109); Benzene-1,3-disulfonic acid 1-amide 3-((2 syn,
3S)-2-hydroxy-4-phenyl-3-(-
3-(S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-amide
(compound 112); Furan-3-sulfonic acid (2 syn,
3S)-2-hydroxy-4-phenyl-3-((-
S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-amide
(compound 113); N-((3-Allyloxycarbonylamino)-(2 syn,
3S)-2-hydroxy-4-phenyl-butyl)--
N-cyclopentylmethyl-4-fluoro-benzenesulfonamide (compound 114);
N-Cyclopentylmethyl-N-((3-ethoxycarbonylamino)-(2 syn,
3S)-2-hydroxy-4-phenyl-butyl)-4-fluoro-benzenesulfonamide (compound
115); 4-Chloro-N-cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tet-
rahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide
(compound 116); 4-Chloro-N-cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-(p-
yridin-3yl-methoxycarbonyl)-butyl)-benzenesulfonamide (compound
118); N-(4-(Cyclopentylmethyl-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydro-
furan-3-yloxycarbonylamino)-butyl)-sulfamoyl)-phenyl)-acetamide
(compound 125); 3-Chloro-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-
-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide (compound
138);
4-Chloro-N-cyclopentylmethyl-N-(2-(syn)-hydroxy-3-((2-oxazolidon-4-(S)-yl-
-methyl)-oxycarbonylamino)-4-phenyl-butyl)-benzenesulfonamide
(compound 139); N-cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrah-
ydrofuran-3-yloxycarbonylamino)-butyl)-4-methoxy-benzenesulfonamide
(compound 140) N-((3-allyloxycarbonylamino)-(2 syn,
3S)-2-hydroxy-4-phenyl-butyl)-N-cyclopentylmethyl-4-methoxy-benzenesulfon-
amide (compound 141); N-Cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-(3-pyridin-3-yl-methoxycarbonylamino)-butyl-4-me-
thoxy-benzenesulfonamide (compound 142); Pyridine-3-sulfonic acid
((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl-
)-isobutyl-amide, trifluoroacetic acid salt (compound 144);
5-Isoxazol-3-yl-thiophene-2-sulfonic acid ((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl-
)-isobutyl-amide (compound 145);
N-(4-((3-(Allyloxycarbonylamino)-(2 syn,
3S)-2-hydroxy-4-phenyl-butyl)-cyclopentylmethylsulfamoyl)-phenyl)-acetami-
de (compound 146); N-(4-(Cyclopentylmethyl-((2 syn,
3S)-2-hydroxy-4-phenyl-3-(pyridin-3-yl-methoxycarbonylamino)-butyl)-sulfa-
moyl)-phenyl)-acetamide (compound 147); N-Cyclopentylmethyl-N-((2
syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl-
)-benzenesulfonamide (compound 148); Pyridine-3-sulfonic acid
cyclopentylmethyl-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran--
3-yloxycarbonylamino)-butyl)-amide (compound 149);
Piperidine-1-sulfonic acid ((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbon-
ylamino)-butyl)-isobutyl-amide (compound 150);
N-4-((2-(syn)-Hydroxy-3-((2-
-methoxymethyl-allyloxycarbonylamino)-4-(S)-phenyl-butyl)-isobutyl-sulfamo-
yl)-phenyl)-acetamide (compound 155);
1-Acetyl-2,3-dihydro-1H-indole-6-sul- fonic acid
((allyloxycarbonylamino)-(2 syn, 3S)-2-hydroxy-4-phenyl-butyl)--
cyclopentylmethyl-amide (compound 156);
1-Acetyl-2,3-dihydro-1H-indole-6-s- ulfonic acid
cyclopentylmethyl-((2 syn, 3S)-2-hydroxy-4-phenyl-3-((S)-tetr-
ahydrofuran-3-yloxycarbonylamino)-butyl)-amide (compound 157);
N-Cyclohexylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofur-
an-3-yloxycarbonylamino)-butyl)-4-methoxy-benzenesulfonamide
(compound 158); N-Cyclohexylmethyl-4-fluoro-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S-
)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide
(compound 159); N-(4-(Cyclohexylmethyl)-((2 syn,
3S)-2-hydroxy-4-phenyl-3-
-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-sulfamoyl-phenyl)-aceta-
mide (compound 160); N-((2 syn,
3S)-2-Hydroxy-4-phenyl-3-(pyridin-4-yl-met-
hoxycarbonylamino)-butyl)-N-isobutyl-4-methoxy-benzenesulfonamide
(compound 163); N-((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((syn)-tetrahydrofura-
n-3-yloxycarbonylamino)-butyl)-N-isobutyl-4-methyl-benzenesulfonamide
(compound 165); N-cyclopentylmethyl-4-hydroxy-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-(pyridin-3-yl-methoxycarbonylamino)-butyl)-benze-
nesulfonamide (compound 166); N-((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((S)-tet-
rahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-4-nitro-benzenesulfon-
amide (compound 167); 4-Amino-N-((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((S)-tet-
rahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide
(compound 168); N-Cyclopentylmethyl-4-hydroxy-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl-
)-benzenesulfonamide (compound 169); N-Cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl-
)-4-nitro-benezensulfonamide (compound 170);
4-Amino-N-cyclopentylmethyl-N- -((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylami-
no)-butyl)-benzenesulfonamide (compound 171);
2,4-Diamino-N-cyclopentylmet- hyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbon-
ylamino)-butyl)-benzenesulfonamide (compound 173); 4-Hydroxy-N-(2
syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl-
)-N-isobutyl-benzenesulfonamide (compound 175);
N-Cyclopentylmethyl-4-fluo- ro-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbony-
lamino)-butyl)-benzenesulfonamide (compound 182);
3,4-Dichloro-N-cyclopent- ylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxyc-
arbonylamino)-butyl)-benzenesulfonamide (compound 183);
Benzyloxycarbonyl-(L)-isoleucine-N-(5-((3-amino-(2 syn,
3S)-2-hydroxy-4-phenyl-butyl)-isobutyl-sulfamoyl)-2-fluoro-phenyl)-acetam-
ide (compound 187); and N-((2 syn,
3S)-4-Cyclohexyl-2-hydroxy-3-((S)-tetra-
hydrofuran-3-yloxycarbonylamino)-butyl)-N-cyclopentylmethyl -4
-methoxy-benzenesulfonamide (compound 195).
10. The compound according to claim 9, said compound being selected
from the group consisting of:
(S)--N-1-(1-(S)-Benzyl-2-(syn)-hydroxyl-3-(isobu-
tyl-(5-pyridin-2-yl-thiophene-2-sulfonyl)-amino)-propyl)-2-((quinoline-2-c-
arbonyl)-amino)-succinamide (compound 12);
(S)--N-1-(1-(S)-Benzyl-3-((4-fl-
uoro-benzenesulfonyl)-isobutyl-amino)-2-(syn)-hydroxy-propyl)-2-((quinolin-
e-2-carbonyl)-amino)-succinamide (compound 14);
(S)--N-1-(3-((4-Acetylamin-
o-3-fluorobenzenesulfonyl)-isobutyl-amino)-(1S,2
syn)-1-benzyl-2-hydroxy-p-
ropyl)-2-((quinoline-2-carbonyl)-amino)-succinamide (compound 15);
(S)--N-1-(3-((Benzo(1,2,5)oxadiazole-4-sulfonyl)-isobutyl-amino)-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinam-
ide (compound 20); N-1-((1S-2
syn)-1-Benzyl-2-hydroxy-3-(1-isobutyl-3,3-di-
methylsulfonylurea)-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide
(compound 21); N-(4-(((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofur-
an-3-yloxycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide
(compound 48); N-((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-
-yloxycarbonylamino)-butyl)-N-isobutyl-4-methoxy-benzenesulfonamide
(compound 100); 4-Chloro-N-cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl-
)-benzenesulfonamide (compound 116); N-Cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl-
)-4-methoxy-benzenesulfonamide (compound 140);
N-Cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-(3-pyridin-3-yl-methoxycarbonylamino)-butyl-
-4-methoxy-benzenesulfonamide (compound 142);
N-Cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)--
butyl)-benzenesulfonamide (compound 148); N-Cyclohexylmethyl-N-((2
syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl-
)-4-methoxy-benzenesulfonamide (compound 158);
N-(4-(Cyclohexylmethyl)-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)--
butyl)-sulfamoyl-phenyl)-acetamide (compound 160);
N-cyclopentylmethyl-4-h- ydroxy-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-(pyridin-3-yl-methoxycarbonylam-
ino)-butyl)-benzenesulfonamide (compound 166); 4-Amino-N-((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl-
)-N-isobutyl-benzenesulfonamide (compound 168);
4-Amino-N-cyclopentylmethy- l-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonyl-
amino)-butyl)-benzenesulfonamide (compound 171);
2,4-Diamino-N-cyclopentyl- methyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycar-
bonylamino)-butyl)-benzenesulfonamide (compound 173);
4-Hydroxy-N-(2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl-
)-N-isobutyl-benzenesulfonamide (compound 175); and N-((2 syn,
3S)-4-Cyclohexyl-2-hydroxy-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-b-
utyl)-N-cyclopentylmethyl-4-methoxy-benzenesulfonamide (compound
195).
11. The compound according to claim 1, wherein said compound has a
molecular weight less than or equal to about 700 g/mol.
12. A compound according to claim 11, wherein said compound has a
molecular weight less than or equal to about 600 g/mol.
13. A compound having the following structural and physicochemical
characteristics: (1) a first and a second hydrogen bond acceptor
moiety, at least one of which is more highly polarizable than a
carbonyl, said moieties being the same or different, and being
capable of hydrogen bonding with the hydrogen atoms of the flap
water molecule of an HIV aspartyl protease when the compound is
bound thereto; (2) substantially hydrophobic moieties which
associate with the P.sub.1 and P.sub.1' binding pockets of said HIV
aspartyl protease when the compound is bound thereto; (3) a third
hydrogen bonding moiety, which may be either a hydrogen bond donor
or acceptor, capable of simultaneously hydrogen bonding to Asp25
and Asp25' of said HIV aspartyl protease when the compound is bound
thereto; (4) an additional occupied volume of space of at least 100
.ANG..sup.3 when the compound is bound to the active site of said
HIV aspartyl protease, said space overlapping with the volume of
space that would be filled by a native substrate of said HIV
aspartyl protease or a nonhyrolyzable isostere thereof; (5) a
deformation energy of binding of the compound to said HIV aspartyl
protease of not greater than 10 kcal/mole; and (6) a neutral or
favorable enthalpic contribution from the sum of all electrostatic
interactions between the compound and the protease when the
compound is bound to said HIV aspartyl protease.
14. The compound according to claim 13, said compound having the
structure of formula XL:
Z.sup.1--Q.sup.1--L.sup.1--M--L.sup.2--Q.sup.2--Z.sup.2 (XL)
wherein: Q.sup.1 and Q.sup.2 are independently hydrogen bond
acceptor moieties capable of binding with the hydrogen atoms of the
flap water molecule of an HIV aspartyl protease, with the proviso
that at least one of Q.sup.1 or Q.sup.2 is more highly polarizable
than a carbonyl; M is a hydrogen bonding moiety, which may be
either a hydrogen bond donor or acceptor, capable of simultaneously
hydrogen bonding to Asp25 and Asp25' of said HIV aspartyl protease;
L.sup.1 and L.sup.2 are independently acyclic or cyclic linker
moieties; and each of Z.sup.1 and Z.sup.2 may be optionally present
and, if present, are independently selected from groups which
occupy a volume of space overlapping with the volume of space that
would be filled by the native substrate of said HIV aspartyl
protease.
15. The compound according to claim 14, wherein at least one of
Q.sup.1 or Q.sup.2 is a substituted sulfonamide.
16. A pharmaceutical composition effective against viral infection
comprising a pharmaceutically effective amount of a compound
according to any one of claims 1-4 and 13-14 and a pharmaceutically
acceptable carrier, adjuvant or vehicle.
17. The pharmaceutical composition according to claim 16, further
comprising an additional anti-viral agent.
18. The use of a compound according to any one of claims 1-4 and
13-14 as a therapeutic agent against viral infection, said virus
requiring an aspartyl protease for an obligatory life cycle
event.
19. The use according to claim 18, wherein said virus is
HIV-1,HIV-2, or HTLV.
20. The use of a compound according to any one of claims 1-4 and
13-14, for inhibiting enzymatic activity in an aspartyl
protease.
21. The use according to claim 20, wherein said aspartyl protease
is HIV protease.
22. A method for preventing HIV infection in a mammal comprising
the step of administering to said mammal a pharmaceutically
effective amount of a pharmaceutical composition according to claim
16 or 17.
23. A method for treating HIV infection in a mammal comprising the
step of administering to said mammal a pharmaceutically effective
amount of a pharmaceutical composition according to claim 16 or
17.
24. The method according to claim 22 or 23, wherein said step of
administering comprises oral administration or administration by
injection.
25. A method for identification, design, or prediction of an HIV
protease inhibitor comprising the steps of: (a) selecting a
candidate compound of defined chemical structure containing a first
and a second hydrogen bond acceptor moiety, at least one of which
is more highly polarizable than a carbonyl, said moieties being the
same or different; a third hydrogen bonding moiety, which may be
either a hydrogen bond donor or acceptor; and at least two
substantially hydrophobic moieties; (b) determining a low-energy
conformation for binding of said compound to the active site of an
HIV aspartyl protease; (c) evaluating the capability of said first
and second hydrogen bond acceptor moieties to form hydrogen bonds
to the flap water molecule of said HIV aspartyl protease when said
compound is bound thereto in said conformation; (d) evaluating the
capability of said substantially hydrophobic moieties to associate
with the P.sub.1 and P.sub.1' binding pockets of said HIV aspartyl
protease when said compound is bound thereto in said conformation;
(e) evaluating the capability of said third hydrogen bonding moiety
to form hydrogen bonds to Asp25 and Asp25' of said HIV aspartyl
protease when said compound is bound thereto in said conformation;
(f) evaluating the overlap of the occupied volume of said compound
when said compound is bound to said HIV aspartyl protease in said
conformation and the occupied volume of a native substrate of HIV
aspartyl protease or a nonhydrolyzable isostere thereof, when said
polypeptide is bound to said HIV aspartyl protease; (g) evaluating
the deformation energy of binding of said compound to said HIV
aspartyl protease; (h) evaluating the enthalpic contribution of the
sum of all electrostatic interactions between said compound and
said HIV aspartyl protease when said compound is bound thereto in
said conformation; and (i) accepting or rejecting said candidate
compound as an HIV protease inhbitor based upon the determinations
and evaluations carried out in steps (b) through (h).
26. An HIV protease inhibitor identified, designed or predicted by
the method according to claim 25.
27. The HIV protease inhibitor according to claim 26, wherein said
inhibitor has the structure of formula XL:
Z.sup.1--Q.sup.1--L.sup.1--M--- L.sup.2--Q.sup.2-Z.sup.2 (XL)
wherein: Q.sup.1 and Q.sup.2 are independently hydrogen bond
acceptor moieties capable of binding with the hydrogen atoms of the
flap water molecule of an HIV aspartyl protease, with the proviso
that at least one of Q.sup.1 or Q.sup.2 is more highly polarizable
than a carbonyl; M is a hydrogen bonding moiety, which may be
either a hydrogen bond donor or acceptor, capable of simultaneously
hydrogen bonding to Asp25 and Asp25' of said HIV aspartyl protease;
L.sup.1 and L.sup.2 are independently acyclic or cyclic linker
moieties; and each of Z.sup.1 and Z.sup.2 may be optionally present
and, if present, are independently selected from groups which
occupy a volume of space overlapping with the volume of space that
would be filled by the native substrate of said HIV aspartyl
protease.
Description
TECHNICAL FIELD OF THE INVENTION
[0001] The present invention relates to a novel class of
sulfonamides which are aspartyl protease inhibitors. In one
embodiment, this invention relates to a novel class of HIV aspartyl
protease inhibitors characterized by specific structural and
physicochemical features. This invention also relates to
pharmaceutical compositions comprising these compounds. The
compounds and pharmaceutical compositions of this invention are
particularly well suited for inhibiting HIV-1 and HIV-2 protease
activity and consequently, may be advantageously used as anti-viral
agents against the HIV-1 and HIV-2 viruses. This invention also
relates to methods for inhibiting the activity of HIV aspartyl
protease using the compounds of this invention and methods for
screening compounds for anti-HIV activity.
BACKGROUND OF THE INVENTION
[0002] The human immunodeficiency virus ("HIV") is the causative
agent for acquired immunodeficiency syndrome ("AIDS")--a disease
characterized by the destruction of the immune system, particularly
of CD4.sup.+ T-cells, with attendant susceptibility to
opportunistic infections--and its precursor AIDS-related complex
("ARC")--a syndrome characterized by symptoms such as persistent
generalized lymphadenopathy, fever and weight loss.
[0003] As in the case of several other retroviruses, HIV encodes
the production of a protease which carries out post-translational
cleavage of precursor polypeptides in a process necessary for the
formation of infectious virions (S. Crawford et al., "A Deletion
Mutation in the 5' Part of the pol Gene of Moloney Murine Leukemia
Virus Blocks Proteolytic Processing of the gag and pol
Polyproteins", J. Virol., 53, p. 899 (1985)). These gene products
include pol, which encodes the virion RNA-dependent DNA polymerase
(reverse transcriptase), an endonuclease, HIV protease, and gag,
which encodes the core-proteins of the virion (H. Toh et al.,
"Close Structural Resemblance Between Putative Polymerase of a
Drosophila Transposable Genetic Element 17.6 and pol gene product
of Moloney Murine Leukemia Virus", EMBO J., 4, p. 1267 (1985); L.
H. Pearl et al., "A Structural Model for the Retroviral Proteases",
Nature, pp. 329-351 (1987); M.D. Power et al., "Nucleotide Sequence
of SRV-1, a Type D Simian Acquired Immune Deficiency Syndrome
Retrovirus", Science, 231, p. 1567 (1986)).
[0004] A number of synthetic anti-viral agents have been designed
to target various stages in the replication cycle of HIV. These
agents include compounds which block viral binding to CD4.sup.+
T-lymphocytes (for example, soluble CD4), and compounds which
interfere with viral replication by inhibiting viral reverse
transcriptase (for example, didanosine and zidovudine (AZT)) and
inhibit integration of viral DNA into cellular DNA (M. S. Hirsh and
R. T. D'Aqulia, "Therapy for Human Immunodeficiency Virus
Infection", N.Eng.J.Med., 328, p. 1686 (1993)). However, such
agents, which are directed primarily to early stages of viral
replication, do not prevent the production of infectious virions in
chronically infected cells. Furthermore, administration of some of
these agents in effective amounts has led to cell-toxicity and
unwanted side effects, such as anemia and bone marrow
suppression.
[0005] More recently, the focus of anti-viral drug design has been
to create compounds which inhibit the formation of infectious
virions by interfering with the processing of viral polyprotein
precursors. Processing of these precursor proteins requires the
action of virus-encoded proteases which are essential for
replication (Kohl, N. E. et al. "Active HIV Protease is Required
for Viral Infectivity" Proc. Natl. Acad. Sci. USA, 85, p. 4686
(1988)). The anti-viral potential of HIV protease inhibition has
been demonstrated using peptidal inhibitors. Such peptidal
compounds, however, are typically large and complex molecules that
tend to exhibit poor bioavailability and are not generally
consistent with oral administration. Accordingly, the need still
exists for compounds that can effectively inhibit the action of
viral proteases, for use as agents for preventing and treating
chronic and acute viral infections.
SUMMARY OF THE INVENTION
[0006] The present invention provides a novel class of compounds,
and pharmaceutically acceptable derivatives thereof, that are
useful as inhibitors of aspartyl proteases, in particular, HIV
aspartyl protease. These compounds can be used alone or in
combination with other therapeutic or prophylactic agents, such as
anti-virals, antibiotics, immunomodulators or vaccines, for the
treatment or prophylaxis of viral infection.
[0007] According to a preferred embodiment, the compounds of this
invention are capable of inhibiting HIV viral replication in human
CD.sub.4.sup.+ T-cells. These compounds are useful as therapeutic
and prophylactic agents to treat or prevent infection by HIV-1 and
related viruses which may result in asymptomatic infection,
AIDS-related complex ("ARC"), acquired immunodeficiency syndrome
("AIDS"), or similar disease of the immune system.
[0008] It is a principal object of this invention to provide a
novel class of sulfonamides which are aspartyl protease inhibitors,
and particularly, HIV aspartyl protease inhibitors. This novel
class of sulfonamides is represented by formula I: 1
[0009] wherein:
[0010] A is selected from the group consisting of H; Het;
--R.sup.1-Het; --R.sup.1--C.sub.1-C.sub.6 alkyl, which may be
optionally substituted with one or more groups selected from the
group consisting of hydroxy, C.sub.1-C.sub.4 alkoxy, Het, --O-Het,
--NR.sup.2--CO--N(R.sup.2)(R.sup.2) and --CO--N(R.sup.2)(R.sup.2);
and --R.sup.1--C.sub.2-C.sub.6 alkenyl, which may be optionally
substituted with one or more groups selected from the group
consisting of hydroxy, C.sub.1-C.sub.4 alkoxy, Het, --O-Het,
--NR.sup.2--CO--N(R.sup.2)(R.sup.2) and
--CO--N(R.sup.2)(R.sup.2);
[0011] each R.sup.1 is independently selected from the group
consisting of --C(O)--, --S(O).sub.2--, --C(O)--C(O)--,
--O--C(O)--, --O--S(O).sub.2, --NR--S(O).sub.2--, --NR--C(O)-- and
--NR--C(O)--C(O)--;
[0012] each Het is independently selected from the group consisting
of C.sub.3-C.sub.7 cycloalkyl; C.sub.5-C.sub.7 cycloalkenyl;
C.sub.6-C.sub.10 aryl; and 5-7 membered saturated or unsaturated
heterocycle, containing one or more heteroatoms selected from N,
N(R.sup.2), O, S and S(O).sub.n, wherein said heterocycle may
optionally be benzofused; and wherein any member of said Het may be
optionally substituted with one or more substituents selected from
the group consisting of oxo, --OR.sup.2, --R.sup.2,
--N(R.sup.2)(R.sup.2), --R.sup.2--OH, --CN, --CO.sub.2R.sup.2,
--C(O)--N(R.sup.2)(R.sup.2), --S(O).sub.2--N(R.sup.2)(R.sup.2),
--N(R.sup.2)--C(O)--R.sub.2, --C(O)--R.sup.2, S(O).sub.n--R.sup.2,
--OCF.sub.3, --S(O).sub.n--Ar, methylenedioxy,
--N(R.sup.2)--S(O).sub.2(R.sup.2), halo, --CF.sub.3, --NO.sub.2, Ar
and --O--Ar;
[0013] each R.sup.2 is independently selected from the group
consisting of H and C.sub.1-C.sub.3 alkyl optionally substituted
with Ar;
[0014] B, when present, is --N(R.sup.2)--C(R.sup.3)
(R.sup.3)--C(O)--;
[0015] x is 0 or 1;
[0016] each R.sup.3 is independently selected from the group
consisting of H, Het, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.3-C.sub.6 cycloalkyl and C.sub.5-C.sub.6
cycloalkenyl, wherein any member of said R.sup.3, except H, may be
optionally substituted with one or more substituents selected from
the group consisting of --OR , --C(O)--NH--R , --S(O) n
N(R.sup.2)(R.sup.2), Het, --CN, --SR.sup.2, --CO.sub.2R.sup.2,
NR.sup.2--C(O)--R.sup.2 each n is independently 1 or 2;
[0017] D and D' are independently selected from the group
consisting of Ar; C.sub.1-C.sub.4 alkyl, which may be optionally
substituted with one or more groups selected from C.sub.3-C.sub.6
cycloalkyl, --OR.sub.2, --R.sup.3, --O--Ar and Ar; C.sub.2-C.sub.4
alkenyl, which may be optionally substituted with one or more
groups selected from the group consisting of C.sub.3-C.sub.6
cycloalkyl, --OR.sup.2, --R.sup.3, --O--Ar and Ar; C.sub.3-C.sub.6
cycloalkyl, which may be optionally substituted with or fused with
Ar; and C.sub.5-C.sub.6 cycloalkenyl, which may be optionally
substituted with or fused with Ar;
[0018] each Ar is independently selected from the group consisting
of phenyl; 3-6 membered carbocyclic ring and 5-6 membered
heterocyclic ring containing one or more heteroatoms selected from
O, N, S, S(O).sub.n and N(R.sup.2), wherein said carbocyclic or
heterocyclic ring may be saturated or unsaturated and optionally
substituted with one or more groups selected from the group
consisting of oxo, --OR.sup.2, --R.sup.2, --N(R.sup.2)(R.sup.2),
--N(R.sup.2)--C(O)--R.sup.2, --R.sup.2--OH, --CN,
--CO.sub.2R.sup.2, --C(O) --N(R.sup.2)(R.sup.2), halo and
--CF.sub.3;
[0019] E is selected from the group consisting of Het; O-Het;
Het-Het; --O--R.sup.2; --NR.sup.2R.sup.3; C.sub.1-C.sub.6 alkyl,
which may be optionally substituted with one or more groups
selected from the group consisting of R.sup.4 and Het;
C.sub.2-C.sub.6 alkenyl, which may be optionally substituted with
one or more groups selected from the group consisting of R.sup.4
and Het; C.sub.3-C.sub.6 saturated carbocycle, which may optionally
be substituted with one or more groups selected from the group
consisting of R.sup.4 and Het; and C.sub.5-C.sub.6 unsaturated
carbocycle, which may optionally be substituted with one or more
groups selected from the group consisting of R.sup.4 and Het;
and
[0020] each R.sup.4 is independently selected from the group
consisting of --OR.sup.2, --C(O)--NHR.sup.2, S(O).sub.2--NHR.sup.2,
halo, --NR.sup.2--C(O)--R.sup.2 and --CN.
[0021] It is a also an object of this invention to provide
pharmaceutical compositions comprising the sulfonamides of formula
I and methods for their use as inhibitors of HIV aspartyl
protease.
[0022] It is a further object of this invention to provide a novel
class of HIV aspartyl protease inhibitor compounds characterized by
the following novel combination of structural and physicochemical
features:
[0023] (1) a first and a second hydrogen bond acceptor moiety, at
least one of which is more highly polarizable than a carbonyl, said
moieties being the same or different, and being capable of hydrogen
bonding with the hydrogen atoms of the flap water molecule of an
HIV aspartyl protease when the compound is bound thereto;
[0024] (2) substantially hydrophobic moieties which associate with
the P.sub.1 and P.sub.1' binding pockets of said HIV aspartyl
protease when the compound is bound thereto;
[0025] (3) a third hydrogen bonding moiety, which may be either a
hydrogen bond donor or acceptor, capable of simultaneously hydrogen
bonding to Asp25 and Asp25' of said HIV aspartyl protease when the
compound is bound thereto;
[0026] (4) an additional occupied volume of space of at least 100
.ANG..sup.3 when the compound is bound to the active site of said
HIV aspartyl protease, said space overlapping with the volume of
space that would be filled by a native substrate of said HIV
aspartyl protease or a nonhyrolyzable isostere thereof;
[0027] (5) a deformation energy of binding of the compound to said
HIV aspartyl protease of not greater than 10 kcal/mole; and
[0028] (6) a neutral or favorable enthalpic contribution from the
sum of all electrostatic interactions between the compound and the
protease when the compound is bound to said HIV aspartyl
protease.
[0029] It is also an object of this invention to provide
pharmaceutical compositions comprising compounds having the
above-mentioned features and methods for their use as inhibitors of
HIV aspartyl protease.
[0030] It is a further object of this invention to provide a method
for identification, design, or prediction of HIV aspartyl protease
inhibitors comprising the steps of:
[0031] (a) selecting a candidate compound of defined chemical
structure containing a first and a second hydrogen bond acceptor
moiety, at least one of which is more highly polarizable than a
carbonyl, said moieties being the same or different; a third
hydrogen bonding moiety, which may be either a hydrogen bond donor
or acceptor; and at least two substantially hydrophobic
moieties;
[0032] (b) determining a low-energy conformation for binding of
said compound to the active site of an HIV aspartyl protease;
[0033] (c) evaluating the capability of said first and second
hydrogen bond acceptor moieties to form hydrogen bonds to the flap
water molecule of said HIV aspartyl protease when said compound is
bound thereto in said conformation;
[0034] (d) evaluating the capability of said substantially
hydrophobic moieties to associate with the P.sub.1 and P.sub.1'
binding pockets of said HIV aspartyl protease when said compound is
bound thereto in said conformation;
[0035] (e) evaluating the capability of said third hydrogen bonding
moiety to form hydrogen bonds to Asp25 and Asp25' of said HIV
aspartyl protease when said compound is bound thereto in said
conformation;
[0036] (f) evaluating the overlap of the occupied volume of said
compound when said compound is bound to said HIV aspartyl protease
in said conformation and the occupied volume of a native substrate
of HIV aspartyl protease or a nonhydrolyzable isostere thereof,
when said polypeptide is bound to said HIV aspartyl protease;
[0037] (g) evaluating the deformation energy of binding of said
compound to said HIV aspartyl protease;
[0038] (h) evaluating the enthalpic contribution of the sum of all
electrostatic interactions between said compound and said HIV
aspartyl protease when said compound is bound thereto in said
conformation; and
[0039] (i) accepting or rejecting said candidate compound as an HIV
protease inhbitor based upon the determinations and evaluations
carried out in steps (b) through (h).
BRIEF DESCRIPTION OF THE DRAWINGS
[0040] FIG. 1 depicts a stereo drawing of a low-energy conformation
of Compound 140, as predicted by computer-modelling.
[0041] FIG. 2 depicts a stereo drawing of the actual crystal
structure of Compound 140, as observed by X-ray
crystallography.
[0042] FIG. 3 depicts a stereo drawing of the correlation between
the predicted (thin line) and observed (thick line) conformation of
Compound 140.
DETAILED DESCRIPTION OF THE INVENTION
[0043] In order that the invention herein described may be more
fully understood, the following detailed description is set forth.
In the description, the following abbreviations are used:
1 Designation Reagent or Fragment Ac acetyl Me methyl Et ethyl Bzl
benzyl Trityl triphenylmethyl Asn D- or L-asparagine Ile D- or
L-isoleucine Phe D- or L-phenylalanine Val D- or L-valine Boc
tert-butoxycarbonyl Cbz benzyloxycarbonyl (carbobenzyloxy) Fmoc
9-fluorenylmethoxycarbonyl DCC dicyclohexylcarbodiimide DIC
diisopropylcarbodiimide EDC
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride HOBt
1-hydroxybenzotriazole HOSu 1-hydroxysuccinimide TFA
trifluoroacetic acid DIEA diisopropylethylamine DBU
1,8-diazabicyclo(5.4.0)undec-7-ene EtOAc ethyl acetate
[0044] The following terms are employed herein:
[0045] Unless expressly stated to the contrary, the terms
"--SO.sub.2--" and "--S(O).sub.2--" as used herein refer to a
sulfone or sulfone derivative (i.e., both appended groups linked to
the S), and not a sulfinate ester.
[0046] For the compounds of formula I, and intermediates thereof,
the stereochemistry of the explicitly shown hydroxyl is defined
relative to D on the adjacent carbon atom, when the molecule is
drawn in an extended zig-zag representation (such as that drawn for
compounds of formula XI, XV, XXII, XXIII and XXXI). If both OH and
D reside on the same side of the plane defined by the extended
backbone of the compound, the stereochemistry of the hydroxyl will
be referred to as "syn". If OH and D reside on opposite sides of
that plane, the stereochemistry of the hydroxyl will be referred to
as "anti".
[0047] The term "heterocyclic" refers to a stable 5-7 membered
monocycle or 8-11 membered bicyclic heterocycle which is either
saturated or unsaturated, and which may be optionally benzofused if
monocyclic. Each heterocycle consists of carbon atoms and from one
to four heteroatoms selected from the group consisting of nitrogen,
oxygen and sulfur. As used herein, the terms "nitrogen and sulfur
heteroatoms" include any oxidized form of nitrogen and sulfur, and
the quaternized form of any basic nitrogen. The heterocyclic ring
may be attached by any heteroatom of the cycle which results in the
creation of a stable structure. Preferred heterocycles defined
above include, for example, benzimidazolyl, imidazolyl,
imidazolinoyl, imidazolidinyl, quinolyl, isoquinolyl, indolyl,
pyridyl, pyrrolyl, pyrrolinyl, pyrazolyl, pyrazinyl, quinoxolyl,
piperidinyl, morpholinyl, thiamorpholinyl, furyl, thienyl,
triazolyl, thiazolyl, .beta.-carbolinyl, tetrazolyl, thiazolidinyl,
benzofuanoyl, thiamorpholinyl sulfone, benzoxazolyl,
oxopiperidinyl, oxopyrroldinyl, oxoazepinyl, azepinyl, isoxazolyl,
tetrahydropyranyl, tetrahydrofuranyl, thiadiazoyl, benzodioxolyl,
thiophenyl, tetrahydrothiophenyl and sulfolanyl.
[0048] The terms "HIV protease" and "HIV aspartyl protease" are
used interchangeably and refer to the aspartyl protease encoded by
the human immunodeficiency virus type 1 or 2. In a preferred
embodiment of this invention, these terms refer to the human
immunodeficiency virus type 1 aspartyl protease.
[0049] The term "hydrophobic" refers to a moiety which tends not to
dissolve readily in water and is often fat-soluble. Hydrophobic
moieties include, but are not limited to, hydrocarbons, such as
alkanes, alkenes, alkynes, cycloalkanes, cycloalkenes, cycloalkynes
and aromatic hydrocarbons, such as aryls, certain saturated and
unsaturated heterocycles and moieties that are substantially
similar to the side chains of hydrophobic natural and unnatural
.alpha.-amino acids, including valine, leucine, isoleucine,
methionine, phenylalanine, .alpha.-amino isobutyric acid,
alloisoleucine, tyrosine, and tryptophan.
[0050] The term "substantially hydrophobic" refers to a hydrophobic
moiety which may optionally contain polar atoms or groups in the
region of the moiety which are solvent exposed when the compound is
bound in the active site of an aspartyl protease.
[0051] The term "linker moiety" refers to a group within a
compound, said group consisting of a backbone of 1-6 atoms selected
from the group consisting of C, N, O, S and P, said backbone being
substituted with, fused to or otherwise associated with a
substantially hydrophobic group capable of associating with the P
or binding pocket of an HIV aspartyl protease when said compound is
bound thereto. In alternative embodiments of this invention, such
linker moieties may optionally be substituted with a group or
groups which occupy a volume of space overlapping with the volume
of space that would be filled by a native substrate of HIV aspartyl
protease or a nonhydrolyzable isostere thereof.
[0052] The term "more highly polarizable than a carbonyl" refers to
a moiety having a polarizability (a) greater than that of a
carbonyl group of a corresponding aldehyde, ketone, ester or amide
moiety.
[0053] The term "pharmaceutically effective amount" refers to an
amount effective in treating HIV infection in a patient. The term
"prophylactically effective amount" refers to an amount effective
in preventing HIV infection in a patient. As used herein, the term
"patient" refers to a mammal, including a human.
[0054] The term "pharmaceutically acceptable carrier or adjuvant"
refers to a non-toxic carrier or adjuvant that may be administered
to a patient, together with a compound of this invention, and which
does not destroy the pharmacological activity thereof.
[0055] As used herein, the compounds of this invention, including
the compounds of formula I, are defined to include pharmaceutically
acceptable derivatives thereof. A "pharmaceutically acceptable
derivative" means any pharmaceutically acceptable salt, ester, or
salt of such ester, of a compound of this invention or any other
compound which, upon administration to a recipient, is capable of
providing (directly or indirectly) a compound of this invention or
an anti-virally active metabolite or residue thereof.
[0056] Pharmaceutically acceptable salts of the compounds of this
invention include those derived from pharmaceutically acceptable
inorganic and organic acids and bases. Examples of suitable acids
include hydrochloric, hydrobromic, sulfuric, nitric, perchloric,
fumaric, maleic, phosphoric, glycollic, lactic, salicylic,
succinic, toluene-p-sulfonic, tartaric, acetic, citric,
methanesulfonic, formic, benzoic, malonic, naphthalene-2-sulfonic
and benzenesulfonic acids. Other acids, such as oxalic, while not
in themselves pharmaceutically acceptable, may be employed in the
preparation of salts useful as intermediates in obtaining the
compounds of the invention and their pharmaceutically acceptable
acid addition salts.
[0057] Salts derived from appropriate bases include alkali metal
(e.g., sodium), alkaline earth metal (e.g., magnesium), ammonium
and N-(C.sub.14 alkyl).sub.4+salts.
[0058] The term "thiocarbamates" refers to compounds containing the
functional group N--SO.sub.2--O.
[0059] The compounds of this invention contain one or more
asymmetric carbon atoms and thus 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. The
explicitly shown hydroxyl is also preferred to be syn to D, in the
extended zig-zag conformation between the nitrogens shown in
compounds of formula I.
[0060] Combinations of substituents and variables envisioned by
this invention are only those that result in the formation of
stable compounds. The term "stable", as used herein, refers to
compounds which possess stability sufficient 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
conditions, for at least a week.
[0061] The compounds of the present invention may be used in the
form of salts derived from inorganic or organic acids. Included
among such acid salts, for example, are the following: acetate,
adipate, alginate, aspartate, benzoate, benzenesulfonate,
bisulfate, butyrate, citrate, camphorate, camphorsulfonate,
cyclopentanepropionate, digluconate, dodecylsulfate,
ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate,
hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide,
hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate,
methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate,
pamoate, pectinate, persulfate, 3-phenylpropionate, picrate,
pivalate, propionate, succinate, tartrate, thiocyanate, tosylate
and undecanoate.
[0062] This invention also envisions the quaternization of any
basic nitrogen-containing groups of the compounds disclosed herein.
The basic nitrogen can be quaternized with any agents known to
those of ordinary skill in the art including, for example, lower
alkyl halides, such as methyl, ethyl, propyl and butyl chloride,
bromides and iodides; dialkyl sulfates including dimethyl, diethyl,
dibutyl and diamyl sulfates; long chain halides such as decyl,
lauryl, myristyl and stearyl chlorides, bromides and iodides; and
aralkyl halides including benzyl and phenethyl bromides. Water or
oil-soluble or dispersible products may be obtained by such
quaternization.
[0063] The novel sulfonamides of this invention are those of
formula I: 2
[0064] wherein:
[0065] A is selected from the group consisting of H; Het;
--R.sup.1-Het; --R.sup.1--C.sub.1-C.sub.6 alkyl, which may be
optionally substituted with one or more groups selected from the
group consisting of hydroxy, C.sub.1-C.sub.4 alkoxy, Het, --O-Het,
--NR.sup.2--CO--N(R.sup.2)(R.sup.2) and --CO--N(R.sup.2)(R.sup.2);
and --R.sup.1--C.sub.2-C.sub.6 alkenyl, which may be optionally
substituted with one or more groups selected from the group
consisting of hydroxy, C.sub.1-C.sub.4 alkoxy, Het, --O-Het, --NR
--CO--N(R.sup.2)(R.sup.2) and --CO--N(R.sup.2)(R.sup.2);
[0066] each R.sup.1 is independently selected from the group
consisting of --C(O)--, --S(O).sub.2--C(O)--C(O)--C(O)--,
O--C(O)--, --O--S(O).sub.2, --NR --S(O).sub.2--, --NR.sup.2--C(O)--
and --NR.sup.2--C(O)--C(O)--;
[0067] each Het is independently selected from the group consisting
of C.sub.3-C.sub.7 cycloalkyl; C.sub.5-C.sub.7 cycloalkenyl;
C.sub.6-C.sub.10 aryl; and 5-7 membered saturated or unsaturated
heterocycle, containing one or more heteroatoms selected from N,
N(R.sup.2), O, S and S(O).sub.n, wherein said heterocycle may
optionally be benzofused; and wherein any member of said Het may be
optionally substituted with one or more substituents selected from
the group consisting of oxo, --OR.sup.2 --R.sup.2.sub.1
--N(R.sup.2)(R.sup.2) --R.sup.2--OH, --CN, --CO.sub.2R.sup.2,
--C(O)--N(R.sup.2)(R.sup.2), --S(O).sub.2--N(R.sup.2)(R.sup.2),
--N(R.sup.2)--C(O)--R.sub.2, --C(O)--R.sup.2, --S(O).sub.n--R,
OCF.sub.3, --S(O).sub.n--Ar, methylenedioxy,
--N(R.sup.2)--S(O).sub.2(R.sup.2), halo, --CF.sub.31 --NO.sub.2, Ar
and --O--Ar;
[0068] each R.sup.2 is independently selected from the group
consisting of H and C.sub.1 -C.sub.3 alkyl optionally substituted
with Ar;
[0069] B, when present, is --N(R.sup.2)--C(R.sup.3)
(R.sup.3)--C(O)--;
[0070] x is 0 or 1;
[0071] each R.sup.3 is independently selected from the group
consisting of H, Het, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.3-C.sub.6 cycloalkyl and C.sub.5-C.sub.6
cycloalkenyl, wherein any member of said R.sup.3, except H, may be
optionally substituted with one or more substituents selected from
the group consisting of --OR.sup.2, --C(O)--NH--R.sup.2,
--S(O).sub.n--N(R.sup.2)(R.sup.2), Het, --CN, --SR.sup.2,
--CO.sub.2R.sup.2, NR.sup.2--C(O)--R.sup.2;
[0072] each n is independently 1 or 2;
[0073] D and D' are independently selected from the group
consisting of Ar; C.sub.1-C.sub.4 alkyl, which may be optionally
substituted with one or more groups selected from C.sub.3-C.sub.6
cycloalkyl, --OR.sub.2, --R.sup.3, --O--Ar and Ar; C.sub.2-C.sub.4
alkenyl, which may be optionally substituted with one or more
groups selected from the group consisting of C.sub.3-C.sub.6
cycloalkyl, --OR.sup.2.sub.1 --R.sup.3, --O--Ar and Ar;
C.sub.3-C.sub.6 cycloalkyl, which may be optionally substituted
with or fused with Ar; and C.sub.5-C.sub.6 cycloalkenyl, which may
be optionally substituted with or fused with Ar;
[0074] each Ar is independently selected from the group consisting
of phenyl; 3-6 membered carbocyclic ring and 5-6 membered
heterocyclic ring containing one or more heteroatoms selected from
O, N, S, S(O) and N(R.sup.2), wherein said carbocyclic or
heterocyclic ring may be saturated or unsaturated and optionally
substituted with one or more groups selected from the group
consisting of oxo, --OR.sup.2.sub.1 --R.sup.2.sub.1
--N(R.sup.2)(R.sup.2), --N(R.sup.2)--C(O)--R.sup.2.sub.1
--R.sup.2--OH, --CN, --CO.sub.2R.sup.2.sub.1
--C(O)--N(R.sup.2)(R.sup.2), halo and --CF.sub.3;
[0075] E is selected from the group consisting of Het; O-Het;
Het-Het; --O--R.sup.3; --NR.sup.2R.sup.3; C.sub.1-C.sub.6 alkyl,
which may be optionally substituted with one or more groups
selected from the group consisting of R.sup.4 and Het;
C.sub.2-C.sub.6 alkenyl, which may be optionally substituted with
one or more groups selected from the group consisting of R.sup.4
and Het; C.sub.3-C.sub.6 saturated carbocycle, which may optionally
be substituted with one or more groups selected from the group
consisting of R.sup.4 and Het; and C.sub.5-C.sub.6 unsaturated
carbocycle, which may optionally be substituted with one or more
groups selected from the group consisting of R.sup.4 and Het;
and
[0076] each R.sup.4 is independently selected from the group
consisting of --OR.sup.2, --C(O)--NHR.sup.2,
--S(O).sub.2-NHR.sup.2, halo, --NR.sup.2C(O)--R.sup.2 and --CN.
[0077] Except where expressly provided to the contrary, as used
herein, the definitions of variables A, R.sup.1--R.sup.4, Het, B,
x, n, D, D', Ar and E are to be taken as they are defined above for
the compounds of formula I.
[0078] According to one embodiment of this invention, a subclass of
compounds are those compounds of formula I, and pharmaceutically
acceptable salts thereof, wherein:
[0079] A is selected from the group consisting of H; --R.sup.1-Het;
--R.sup.1--C.sub.1-C.sub.6 alkyl, which may be optionally
substituted with one or more groups selected from the group
consisting of hydroxy, C.sub.1-C.sub.4 alkoxy, Het and --O-Het; and
--R.sup.1--C.sub.2-C.sub.6 alkenyl, which may be optionally
substituted with one or more groups selected from hydroxy,
C.sub.1-C.sub.4 alkoxy, Het and --O-Het;
[0080] each R.sup.1 is independently selected from the group
consisting of --C(O)--, --S(O).sub.2--, --C(O)--C(O)--, --O--CO--,
--S(O).sub.2-- and --NR.sup.2--S(O).sub.2--;
[0081] each Het is independently selected from the group consisting
of C.sub.3-C.sub.7 cycloalkyl; C.sub.5-C.sub.7 cycloalkenyl;
C.sub.6-C.sub.10 aryl; and 5-7 membered saturated or unsaturated
heterocycle, containing one or more heteroatoms selected from N, O
and S, which may optionally be benzofused; wherein any member of
said Het may be optionally substituted with one or more
substituents selected from the group consisting of oxo, --OR.sup.2,
--R.sup.2, --N(R.sup.2).sub.2, --R.sup.2--OH, --CN,
--CO.sub.2R.sup.2, --C(O)--N(R.sup.2).sub.2 and
--S(O).sub.2--N(R.sup.2).sub.2;
[0082] each R.sup.2 is independently selected from the group
consisting of H and C.sub.1-C.sub.3 alkyl;
[0083] B, when present, is --NH--CH(R.sup.3)--C(O)--;
[0084] x is 0 or 1;
[0085] R.sup.3 is selected from the group consisting of Het,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.3-C.sub.6
cycloalkyl and C.sub.5-C.sub.6 cycloalkenyl, wherein any member of
said R.sup.3 may be optionally substituted with one or more
substituents selected from the group consisting of --OR.sup.2,
--C(O)--NH--R.sub.2, --S(O).sub.n--N(R.sup.2).sub.2, Het and
--CN;
[0086] n is 1 or 2;
[0087] D and D' are independently selected from the group
consisting of Ar; C.sub.1-C.sub.4 alkyl, which may be optionally
substituted with C.sub.3-C.sub.6 cycloalkyl or Ar; C.sub.2-C.sub.4
alkenyl, which may be optionally substituted with C.sub.3-C.sub.6
cycloalkyl or Ar; C.sub.3-C.sub.6 cycloalkyl, which may be
optionally substituted or fused with Ar; and C.sub.5-C.sub.6
cycloalkenyl, which may be optionally substituted or fused with Ar;
with the proviso that when D is attached to N, D may not be methyl
or C.sub.2 alkenyl;
[0088] Ar is selected from the group consisting of phenyl; 3-6
membered carbocyclic ring and 5-6 membered heterocyclic ring
containing one or more heteroatoms selected from O, N and S,
wherein said carbocyclic or heterocyclic ring may be saturated or
unsaturated and optionally substituted with one or more groups
selected from the group consisting of oxo, --OR.sup.2, --R.sup.2,
--N(R.sup.2).sub.2, --N(R.sup.2)--C(O)R.sup.2- , --R.sup.2--OH,
--CN, --CO.sub.2R.sup.2, --C(O)--N(R.sup.2).sub.2; halo and
--CF.sub.3;
[0089] E is selected from the group consisting of Het;
--O--R.sup.3; --NR.sup.2R.sup.5; C.sub.1-C.sub.6 alkyl, which may
be optionally substituted with one or more R.sup.4 or Het;
C.sub.2-C.sub.6 alkenyl, which may be optionally substituted with
one or more R.sup.4 or Het; C.sub.3-C.sub.6 saturated carbocycle,
which may optionally be substituted with one or more R.sup.4 or
Het; and C.sub.5-C.sub.6 unsaturated carbocycle, which may
optionally be substituted with one or more R.sup.4 or Het;
[0090] each R.sup.4 is independently selected from the group
consisting of --OR.sup.2,
--C(O)--NHR.sup.2,--S(O).sub.2--NHR.sup.2, halo and --CN; and
[0091] each R.sup.5 is independently selected from the group
consisting of H and R.sup.3, with the proviso that at least one
R.sup.5 is not H.
[0092] A preferred subclass of compounds of this invention are
those compounds of formula I having a molecular weight of less than
about 700 g/mole. More preferably, the subclass of compounds of
formula I have a molecular weight of less than about 600
g/mole.
[0093] Other preferred subclasses of this invention are those
compounds of formulas XXII, XXIII and XXXI: 3
[0094] wherein A, R.sup.3, Het, D, D', x and E are as defined above
for compounds of formula I. For ease of reference, the two R.sup.3
moieties present in formula XXXI have been labeled R.sup.3 and
R.sup.3'.
[0095] For compounds of formula XXII, most preferred compounds are
those wherein A is R.sup.1-Het and D' is C.sub.1-C.sub.3 alkyl or
C.sub.3 alkenyl, wherein said alkyl or alkenyl may optionally be
substituted with one or more groups selected from the group
consisting of C.sub.3-C.sub.6 cycloalkyl, --OR.sub.2, --O--Ar and
Ar (with all other variables being defined as above for compounds
of formula I). For compounds of formula XXIII, most preferred
compounds are those wherein R.sup.3 is C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.5 -C.sub.6 cycloalkyl,
C.sub.5-C.sub.6 cycloalkenyl or a 5-6 membered saturated or
unsaturated heterocycle, wherein any member of said R.sup.3 may be
optionally substituted with one or more substituents selected from
the group consisting of --OR.sup.2.sub.1 --C(O)--NH--R.sup.2,
--S(O).sub.nN(R.sup.2)(R.sup.2), Het, --CN, --SR.sup.2,
--C(O).sub.2R.sup.2 and NR.sup.2--C(O)--R.sup.2 and D' is
C.sub.1-C.sub.3 alkyl or C.sub.3 alkenyl, wherein said alkyl or
alkenyl may optionally be substituted with one or more groups
selected from the group consisting of C.sub.3-C.sub.6 cycloalkyl,
--OR.sub.2, --O--Ar and Ar (with all other variables being defined
as above for compounds of formula I).
[0096] For compounds of formula XXXI, most preferred compounds are
those wherein A is R.sup.1-Het, each R.sup.3 is independently
C.sub.1-C.sub.6 alkyl which may be optionally substituted with a
substituent selected from the group consisting of --OR.sup.2,
--C(O)--NH--R.sup.2, --S(O).sub.nN(R.sup.2)(R.sup.2), Het. --CN,
--SR.sup.2, --CO.sub.2R.sup.2 and --NR.sup.2--C(O)--R.sup.2; D' is
C.sub.1-C.sub.4 alkyl, which may be optionally substituted with a
group selected from the group consisting of C.sub.3-C.sub.6
cycloalkyl, --OR.sup.2, --O--Ar; and E is Het, Het-Het and
--NR.sup.2R.sup.3.
[0097] Sulfonamides of this invention include the following
specific compounds contained in Tables I-VI. In Tables I-IV and VI,
A is attached through the right-most bond, unless otherwise
expressly noted. All other substituents in Tables I-VI are attached
via the left-most bond, unless otherwise expressly noted.
2TABLE I 4 COMPOUND A R.sup.3 D' E 1 5 6 7 8 2 9 10 11 12 3 13 14
15 16 4 17 18 19 20 5 21 22 23 24 6 25 26 27 28 7 29 30 31 32 8 33
34 35 CH.sub.3 9 36 37 38 39 10 40 41 42 43 11 44 45 46 47 12 48 49
50 51 13 52 53 54 55 14 56 57 58 59 15 60 61 62 63 16 64 65 66 67
17 68 69 70 71 18 72 73 74 75 19 76 77 78 79 20 80 81 82 83 21 84
85 86 87 22 88 89 90 91 23 92 93 94 95 24 96 97 98 99 25 100 101
102 103 26 104 105 106 107
[0098]
3TABLE II 108 COMPOUND A D' E 27 109 110 111 28 112 113 114 29 115
116 117 30 118 119 120 31 121 122 123 32 124 125 126 33 127 128 129
34 130 131 132 35 133 134 135 36 136 137 138 37 139 140 141 38 142
143 144 39 145 146 147 40 148 149 150 41 151 152 153 42 154 155 156
43 157 158 159 44 160 161 162 45 163 164 165 46 166 167 168 47 169
170 171 48 172 173 174 49 175 176 177 50 178 179 180 51 181 182 183
52 184 185 186 53 187 188 189 54 190 191 192 55 193 194 195 56 196
197 198 57 199 200 201 58 202 203 204 59 205 206 207 60 208 209 210
61 211 212 213 62 214 215 216 63 217 218 219 64 220 221 222 65 223
224 225 66 226 227 228 67 229 230 231 68 232 233 234 69 235 236 237
70 238 239 240 71 241 242 243 72 244 245 246 73 247 248 249 74 250
251 252 75 253 254 255 76 256 257 258 77 259 260 261 78 262 263 264
79 265 266 267 80 268 269 270 81 271 272 273 82 274 275 276 83 277
278 279 84 280 281 282 85 283 284 285 86 286 287 288 87 289 290 291
88 292 293 294 89 295 296 297 90 298 299 300 91 301 302 303 92 304
305 306 93 307 308 309 94 310 311 312 95 313 314 315 96 316 317 318
97 319 320 321 98 322 323 324 99 325 326 327 100 328 329 330 101
331 CH.sub.3 332 102 333 334 335 103 336 337 338 104 339 340 341
105 342 343 344 106 345 346 347 107 348 349 350 108 351 352 353 109
354 355 356 110 357 358 359 111 360 361 362 112 363 364 365 113 366
367 368 114 369 370 371 115 372 373 374 116 375 376 377 117 378 379
380 118 381 382 383 119 384 385 386 120 387 388 389 121 390 391 392
122 393 394 395 123 396 397 398 124 399 400 401 125 402 403 404 126
405 CH.sub.3 406 127 407 CH.sub.3 408 128 409 CH.sub.3 410 129 411
412 413 130 414 415 416 131 417 418 419 132 420 421 422 133 423 424
425 134 426 427 428 135 429 430 431 136 432 433 434 137 435 436 437
138 438 439 440 139 441 442 443 140 444 445 446 141 447 448 449 142
450 451 452 143 453 454 455 144 456 457 458 145 459 460 461 146 462
463 464 147 465 466 467 148 468 469 470 149 471 472 473 150 474 475
476 151 477 478 479 152 480 481 482 153 483 484 485 154 486 487 488
155 489 490 491 156 492 493 494 157 495 496 497 158 498 499 500 159
501 502 503 160 504 505 506 161 507 508 509 162 510 511 512 163 513
514 515 164 516 517 518 165 519 520 521 166 522 523 524 167 525 526
527 168 528 529 530 169 531 532 533 170 534 535 536 171 537 538 539
172 540 541 542 173 543 544 545 174 546 547 548 175 549 550 551 176
552 553 554 177 555 556 557 178 558 559 560 179 561 562 563 180 564
565 566 181 567 568 569 182 570 571 572 183 573 574 575 184 576 577
578 185 579 580 581
[0099]
4TABLE III 582 COMPOUND A D' E 186 583 584 585 187 586 587 588 188
589 590 591
[0100]
5TABLE IV 592 COMPOUND A 189 593 190 594
[0101]
6TABLE V 595 COM- POUND E 191 596 192 597 193 598 194 599
[0102]
7TABLE VI 600 COMPOUND D D' 195 601 602 196 603 604
[0103] Preferred compounds of this invention are*: * As can be
appreciated by those of ordinary skill in the art, many different
conventions are used in naming chemical compounds. Because of
possible discrepencies in the art of chemical nomenclature, the
structures shown in Tables I-VI herein are controlling for the
definition of compounds 1-195 of this invention.
[0104]
(S)--N-1-(3-((3-Acetylamino-4-fluorobenzenesulfonyl)-benzyl-amino)--
(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-suc-
cinamide and
(S)--N-1-(3-((4-Acetylamino-3-fluorobenzenesulfonyl)-benzyl-a-
mino)-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amin-
o)-succinamide (compounds 2);
[0105]
(S)--N-1-(3-((5-Acetylamino-3-methyl-thiophene-2-sulfonyl)-benzyl-a-
mino)-(1S,2
syn)-1-benzyl-2-hydroxypropyl)-2-((quinoline-2-carbonyl)-amino-
)-succinamide (compound 5);
[0106]
(S)--N-1-(1-Benzyl-3-(benzyl-(5-isoxazol-3-yl-thiophene-2-sulfonyl)-
-amino)-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-am-
ino)-succinamide (compound 6);
[0107]
(S)--N-1-(3-((Benzo(1,2,5)oxadiazole-4-sulfonyl)-benzyl-amino)-(1S,-
2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succina-
mide (compound 9);
[0108]
N-1-(1-(S)-Benzyl-3-(benzyl-(3-sulfamoylbenzenesulfonyl)-amino)-2-(-
syn)-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide
(compound 10);
[0109]
(S)--N-1-(1-(S)-Benzyl-2-(syn)-hydroxyl-3-(isobutyl-(5-pyridin-2-yl-
-thiophene-2-sulfonyl)-amino)-propyl)-2-((quinoline-2-carbonyl)-amino)-suc-
cinamide (compound 12);
[0110]
(S)--N-1-(3-((4-Benzenesulfonyl-thiophene-2-sulfonyl)-isobutyl-amin-
o)-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)--
succinamide (compound 13);
[0111]
(S)--N-1-(1-(S)-Benzyl-3-((4-fluorobenzenesulfonyl)-isobutyl-amino)-
-2-(syn)-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide
(compound 14);
[0112]
(S)--N-1-(3-((4-Acetylamino-3-fluorobenzenesulfonyl)-isobutyl-amino-
)-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-s-
uccinamide (compound 15);
[0113]
(S)--N-1-(3-((3-Acetylamino-4-fluorobenzenesulfonyl)-isobutyl-amino-
)-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-s-
uccinamide (compound 16);
[0114]
(S)--N-1-(1-(S)-Benzyl-3-((4-acetylamino-benzenesulfonyl)-isobutyl--
amino)-2-(syn)-hydroxypropyl)-2-((quinoline-2-carbonyl)-amino)-succinamide
(compound 17);
[0115]
(S)--N-1-(3-((5-Acetylamino-3-methyl-thiophene-2-sulfonyl)-isobutyl-
-amino)-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-am-
ino)-succinamide (compound 18);
[0116]
(S)--N-1-(3-((3-Acetylamino-benzenesulfonyl)-isobutyl-amino)-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinam-
ide (compound 19);
[0117]
(S)--N-1-(3-((Benzo(1,2,5)oxadiazole-4-sulfonyl)-isobutyl-amino)-(1-
S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succi-
namide (compound 20);
[0118] N-1-((1S-2
syn)-1-Benzyl-2-hydroxy-3-(1-isobutyl-3,3-dimethylsulfon-
ylurea)-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide
(compound 21);
[0119]
N-1-(3-((4-Acetylamino-benzenesulfonyl)-isobutyl-amino)-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-(pyridin-2-yl-methoxycarbonyl)-succinam-
ide (compound 22);
[0120]
N-1-(3-((4-Acetylamino-benzenesulfonyl)-isobutyl-amino)-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-(pyridin-4-yl-methoxycarbonyl)-succinam-
ide (compound 23);
[0121] N-1-(3-((4-Fluoro-benzenesulfonyl)-isobutyl-amino)-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-(pyridin-2-yl-methoxycarbonyl)-succinam-
ide (compound 26);
[0122] 4-Fluoro-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-
-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide (compound
35);
[0123] 3,4-Dichloro-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofur-
an-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide
(compound 37);
[0124] N-(4-(((2 syn,
3S)-2-Hydroxy-4-phenyl-3-(pyridin-3-yl-methoxycarbon-
ylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide (compound
44);
[0125] 2,4-Dimethyl-thiazole-5-sulfonic
acid-(1,1-dimethyl-ethoxycarbonyla- mino)-(2 syn,
3S)-2-hydroxy-4-phenyl-butyl)-isobutyl-amide (compound 46);
[0126] N-(4-(((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-ylox-
ycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide
(compound 48);
[0127] 4-Fluoro-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((R)-tetrahydrofuran-3-
-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide and
4-Fluoro-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((R)-tetrahydrofuran-3-yloxy-
carbonylamino)-butyl)-N-isobutyl-benzenesulfonamide (compounds
52);
[0128] Benzo(1,2,5)oxadiazole-5-sulfonic acid ((2 syn,
3S)-2-hydroxy-4-phenyl-3-(pyridin-3-yl-methoxycarbonylamino)-butyl)-isobu-
tylamide (compound 66);
[0129] N-(4-(((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((R)-tetrahydrofuran-3-ylox-
ycarbonylamino)-butyl)-isobutyl-sulfamoyl-phenyl)-acetamide and
N-(4-(((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)--
butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide (compounds 86);
[0130] N-(2-Fluoro-5-(((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofur-
an-3-yloxycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide
(compound 88);
[0131] N-(3-(((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-ylox-
ycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide
(compound 91);
[0132] 4-Fluoro-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((R)-tetrahydrofuran-3-
-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide (compound
93);
[0133]
N-(4-(((syn)-2-Hydroxy-(S)-4-phenyl-3-((tetrahydro-furan-(R)-3-yl)--
oxycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide
(compound 94);
[0134] 4-Fluoro-N-(2 syn,
3S)-2-hydroxy-4-phenyl-3-((tetrahydro-furan-(R)--
3-ylmethoxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide and
4-Fluoro-N-(2 syn,
3S)-2-hydroxy-4-phenyl-3-((tetrahydro-furan-(S)-3-ylme-
thoxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide (compounds
97);
[0135] 4-Fluoro-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-(pyridin-3-yl-methoxyc-
arbonylamino)-butyl)-N-isobutyl-benzenesulfonamide (compound
98);
[0136] 4-Chloro-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-
-yloxycarbonylamino)-butyl)-isobutyl-benzenesulfonamide (compound
99);
[0137] N-((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycar-
bonylamino)-butyl)-N-isobutyl-4-methoxy-benzenesulfonamide
(compound 100);
[0138]
4-Fluoro-N-(2-(syn)-hydroxy-3-((2-oxazolidon-(S)-4-yl)-methoxycarbo-
nylamino)-4-(S)-phenyl-butyl)-N-isobutyl-benzenesulfonamide
(compound 109);
[0139] Benzene-1,3-disulfonic acid 1-amide 3-((2 syn,
3S)-2-hydroxy-4-phenyl-3-(3-(S)-tetrahydrofuran-3-yloxycarbonylamino)-but-
yl)-isobutyl-amide (compound 112);
[0140] Furan-3-sulfonic acid (2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahy-
drofuran-3-yloxycarbonylamino)-butyl)-isobutyl-amide (compound
113);
[0141] N-((3-Allyloxycarbonylamino)-(2 syn,
3S)-2-hydroxy-4-phenyl-butyl)--
N-cyclopentylmethyl-4-fluorobenzenesulfonamide (compound 114);
[0142] N-Cyclopentylmethyl-N-((3-ethoxycarbonylamino)-(2 syn,
3S)-2-hydroxy-4-phenyl-butyl)-4-fluorobenzenesulfonamide (compound
115);
[0143] 4-Chloro-N-cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((-
S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide
(compound 116);
[0144] 4-Chloro-N-cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-(p-
yridin-3yl-methoxycarbonyl)-butyl)-benzenesulfonamide (compound
118);
[0145] N-(4-(Cyclopentylmethyl-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetr-
ahydrofuran-3-yloxycarbonylamino)-butyl)-sulfamoyl)-phenyl)-acetamide
(compound 125);
[0146] 3-Chloro-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-
-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide (compound
138);
[0147]
4-Chloro-N-cyclopentylmethyl-N-(2-(syn)-hydroxy-3-((2-oxazolidon-4--
(S)-yl-methyl)-oxycarbonylamino)-4-phenyl-butyl)-benzenesulfonamide
(compound 139);
[0148] N-cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrah-
ydrofuran-3-yloxycarbonylamino)-butyl)-4-methoxy-benzenesulfonamide
(compound 140);
[0149] N-((3-allyloxycarbonylamino)-(2 syn,
3S)-2-hydroxy-4-phenyl-butyl)--
N-cyclopentylmethyl-4-methoxy-benzenesulfonamide (compound
141);
[0150] N-Cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-(3-pyridin--
3-yl-methoxycarbonylamino)-butyl-4-methoxy-benzenesulfonamide
(compound 142);
[0151] Pyridine-3-sulfonic acid ((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tet-
rahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-amide,
trifluoroacetic acid salt (compound 144);
[0152] 5-Isoxazol-3-yl-thiophene-2-sulfonic acid ((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl-
)-isobutyl-amide (compound 145);
[0153] N-(4-((3-(Allyloxycarbonylamino)-(2 syn,
3S)-2-hydroxy-4-phenyl-but-
yl)-cyclopentylmethylsulfamoyl)-phenyl)-acetamide (compound
146);
[0154] N-(4-(Cyclopentylmethyl-((2 syn,
3S)-2-hydroxy-4-phenyl-3-(pyridin--
3-yl-methoxycarbonylamino)-butyl)-sulfamoyl)-phenyl)-acetamide
(compound 147);
[0155] N-Cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrah-
ydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide (compound
148);
[0156] Pyridine-3-sulfonic acid cyclopentylmethyl-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl-
)-amide (compound 149);
[0157] Piperidine-1-sulfonic acid ((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-t-
etrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-amide
(compound 150);
[0158]
N-4-((2-(syn)-Hydroxy-3-((2-methoxymethyl-allyloxycarbonylamino)-4--
(S)-phenyl-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide (compound
155);
[0159] 1-Acetyl-2,3-dihydro-1H-indole-6-sulfonic acid
((allyloxycarbonylamino)-(2 syn,
3S)-2-hydroxy-4-phenyl-butyl)-cyclopenty- lmethyl-amide (compound
156);
[0160] 1-Acetyl-2,3-dihydro-1H-indole-6-sulfonic acid
cyclopentylmethyl-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran--
3-yloxycarbonylamino)-butyl)-amide (compound 157);
[0161] N-Cyclohexylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahy-
drofuran-3-yloxycarbonylamino)-butyl)-4-methoxy-benzenesulfonamide
(compound 158);
[0162] N-Cyclohexylmethyl-4-fluoro-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S-
)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide
(compound 159);
[0163] N-(4-(Cyclohexylmethyl)-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetr-
ahydrofuran-3-yloxycarbonylamino)-butyl)-sulfamoyl-phenyl)-acetamide
(compound 160);
[0164] N-((2 syn,
3S)-2-Hydroxy-4-phenyl-3-(pyridin-4-yl-methoxycarbonylam-
ino)-butyl)-N-isobutyl-4-methoxy-benzenesulfonamide (compound
163);
[0165] N-((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((syn)-tetrahydrofuran-3-yloxyc-
arbonylamino)-butyl)-N-isobutyl-4-methyl-benzenesulfonamide
(compound 165);
[0166] N-cyclopentylmethyl-4-hydroxy-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-(-
pyridin-3-yl-methoxycarbonylamino)-butyl)-benzenesulfonamide
(compound 166);
[0167] N-((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycar-
bonylamino)-butyl)-N-isobutyl-4-nitro-benzenesulfonamide (compound
167);
[0168] 4-Amino-N-((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3--
yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide (compound
168);
[0169] N-Cyclopentylmethyl-4-hydroxy-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-(-
(S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide
(compound 169);
[0170] N-Cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrah-
ydrofuran-3-yloxycarbonylamino)-butyl)-4-nitro-benezensulfonamide
(compound 170);
[0171] 4-Amino-N-cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S-
)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide
(compound 171);
[0172] 2,4-Diamino-N-cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-
-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide
(compound 173);
[0173] 4-Hydroxy-N-(2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-
-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide (compound
175);
[0174] N-Cyclopentylmethyl-4-fluoro-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((-
S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide
(compound 182);
[0175] 3,4-Dichloro-N-cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl--
3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide
(compound 183); and
[0176] Benzyloxycarbonyl-(L)-isoleucine-N-(5-((3-amino-(2 syn,
3S)-2-hydroxy-4-phenyl-butyl)-isobutyl-sulfamoyl)-2-fluoro-phenyl)-acetam-
ide (compound 187);
[0177] N-((2 syn,
3S)-4-Cyclohexyl-2-hydroxy-3-((S)-tetrahydrofuran-3-ylox-
ycarbonylamino)-butyl)-N-cyclopentylmethyl-4-methoxy-benzenesulfonamide
(compound 195).
[0178] More preferred compounds of this invention are:
[0179]
(S)--N-1-(1-(S)-Benzyl-2-(syn)-hydroxyl-3-(isobutyl-(5-pyridin-2-yl-
-thiophene-2-sulfonyl)-amino)-propyl)-2-((quinoline-2-carbonyl)-amino)-suc-
cinamide (compound 12);
[0180]
(S)--N-1-(1-(S)-Benzyl-3-((4-fluorobenzenesulfonyl)-isobutyl-amino)-
-2-(syn)-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide
(compound 14);
[0181]
(S)--N-1-(3-((4-Acetylamino-3-fluorobenzenesulfonyl)-isobutyl-amino-
)-(1S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-s-
uccinamide (compound 15);
[0182]
(S)--N-1-(3-((Benzo(1,2,5)oxadiazole-4-sulfonyl)-isobutyl-amino)-(1-
S,2
syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succi-
namide (compound 20);
[0183] N-1-((1S-2
syn)-1-Benzyl-2-hydroxy-3-(1-isobutyl-3,3-dimethylsulfon-
ylurea)-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide
(compound 21);
[0184] N-(4-(((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-ylox-
ycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide
(compound 48);
[0185] N-((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycar-
bonylamino)-butyl)-N-isobutyl-4-methoxy-benzenesulfonamide
(compound 100);
[0186] 4-Chloro-N-cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((-
S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide
(compound 116);
[0187] N-Cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrah-
ydrofuran-3-yloxycarbonylamino)-butyl)-4-methoxy-benzenesulfonamide
(compound 140);
[0188] N-Cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-(3-pyridin--
3-yl-methoxycarbonylamino)-butyl-4-methoxy-benzenesulfonamide
(compound 142);
[0189] N-Cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrah-
ydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide (compound
148);
[0190] N-Cyclohexylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahy-
drofuran-3-yloxycarbonylamino)-butyl)-4-methoxy-benzenesulfonamide
(compound 158);
[0191] N-(4-(Cyclohexylmethyl)-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetr-
ahydrofuran-3-yloxycarbonylamino)-butyl)-sulfamoyl-phenyl)-acetamide
(compound 160);
[0192] N-cyclopentylmethyl-4-hydroxy-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-(-
pyridin-3-yl-methoxycarbonylamino)-butyl)-benzenesulfonamide
(compound 166);
[0193] 4-Amino-N-((2 syn,
3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3--
yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide (compound
168);
[0194] 4-Amino-N-cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-((S-
)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide
(compound 171);
[0195] 2,4-Diamino-N-cyclopentylmethyl-N-((2 syn,
3S)-2-hydroxy-4-phenyl-3-
-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide
(compound 173);
[0196] 4-Hydroxy-N-(2 syn,
3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-
-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide (compound
175); and
[0197] N-((2 syn,
3S)-4-Cyclohexyl-2-hydroxy-3-((S)-tetrahydrofuran-3-ylox-
ycarbonylamino)-butyl)-N-cyclopentylmethyl-4-methoxy-benzenesulfonamide
(compound 195).
[0198] The sulfonamides of this invention may be synthesized using
conventional techniques. Advantageously, these compounds are
conveniently synthesized from readily available starting
materials.
[0199] The compounds of this invention are among the most readily
synthesized HIV protease inhibitors known. Previously described HIV
protease inhibitors often contain four or more chiral centers,
numerous peptide linkages and/or require air-sensitive reagents
(such as organometallic complexes) to effect their synthesis. The
relative ease with which the compounds of this invention can be
synthesized represents an enormous advantage in the large scale
production of these compounds.
[0200] In general, sulfonamides of formula I are conveniently
obtained from .alpha.-amino acid derivatives having the general
formula A--(B).sub.x--NH--CH(D)--COOH, wherein A, B, X and D are
defined as above for the compounds of formula I. Such .alpha.-amino
acid derivatives are often commercially available or may be
conveniently prepared from commercially available .alpha.-amino
acid derivatives using known techniques. See, for example, T. W.
Greene and P. G. M. Wuts, "Protective Groups in Organic Synthesis",
2nd Ed., John Wiley and Sons (1991). Although this invention
envisions the use of racemic mixtures of such starting materials,
when x=0, a single enantiomer in the S configuration is
preferred.
[0201] Using known techniques, the .alpha.-amino acid derivative of
general formula A--(B)--NH--CH(D)--COOH may be readily converted to
an amino ketone derivative of general formula
A--(B).sub.x--NH--CH(D)--CO--C- H.sub.2--X, wherein X is a leaving
group which suitably activates the .alpha.-carbon (i.e., makes the
methylene susceptible to nucleophilic attack). Suitable leaving
groups are well known in the art and include halides and
sulfonates, such as methanesulfonate, trifluoromethanesulfona- te
or 4-toluenesulfonate X may also be a hydroxyl which is converted
in situ to a leaving group (e.g. by treatment with a trialkyl- or
triarylphosphine in the presence of a dialkylazodicarboxylate).
Methods for the formation of such amino ketone derivatives also are
well known to those of skill in the art (see, for example, S. J.
Fittkau, J. Prakt. Chem., 315, p. 1037 (1973)). Alternatively,
certain amino ketone derivatives are commercially available (e.g.,
from Bachem Biosciences, Inc., Philadelphia, Pa.).
[0202] The amino ketone derivative may then be reduced to the
corresponding amino alcohol, represented by the formula
A--(B).sub.x--NH--CH(D)--CH(OH)--CH.sub.2--X. Many techniques for
reduction of amino ketone derivatives such as
A--(B).sub.x--NH--CH(D)--CO- --CH.sub.2--X are well known to those
of ordinary skill in the art (Larock, R. C. "Comprehensive Organic
Transformations", pp. 527-547, VCH Publishers, Inc..COPYRGT. 1989
and references cited therein). A preferred reducing agent is sodium
borohydride. The reduction reaction is conducted at a temperature
of from about -40.degree. C. to about 40.degree. C. (preferably, at
about OC to about 20.degree. C.), in a suitable solvent system such
as, for example, aqueous or neat tetrahydrofuran or a lower
alcohol, such as methanol or ethanol. Although this invention
envisions both stereospecific and non-stereospecific reduction of
the amino ketone derivative
A--(B).sub.x--NH--CH(D)--CO--CH.sub.2--X, stereoselective reduction
is preferred. Stereoselective reduction may be accomplished by use
of chiral reagents known in the art. In the present invention,
stereoselective reduction may be conveniently achieved, for
instance, under non-chelating reducing conditions, where chiral
induction of the newly formed hydroxyl group is set by the
stereochemistry of the D group (i.e., Felkin-Ahn addition of
hydride). We particularly prefer stereoselective reductions wherein
the resulting hydroxyl is syn to D. We have found that when the
hydroxyl group is syn to D, the final sulfonamide product is an HIV
protease inhibitor of higher potency than the anti
diastereomer.
[0203] The hydroxyl group of the amino alcohol may optionally be
protected by any known oxygen protecting group (such as
trialkylsilyl, benzyl, or alkyloxymethyl) to yield a protected
amino alcohol having the formula
A--(B).sub.x--NH--CH(D)--C(OR.sup.6)--CH.sub.2--X, wherein R.sup.6
is H or any suitable hydroxy protecting group. Several useful
protecting groups are described in T. W. Greene and P. G. M. Wuts,
Protective Groups in Organic Synthesis, 2d Ed., John Wiley and Sons
(1991).
[0204] The amino alcohol may then be reacted with a nucleophilic
amine compound to form an intermediate of formula III: 605
[0205] wherein D and R.sup.6 are as described above, and L is
either D' (as described for compounds of formula I) or
hydrogen.
[0206] In a particularly advantageous synthetic scheme,
simultaneous activation of the methylene and protection of the
alcohol may be accomplished by forming an N-protected amino epoxide
from the oxygen and its adjacent methylene to give an intermediate
of formula II: 606
[0207] wherein A, B and D are as defined above for compounds of
formula I. Suitable solvent systems for preparing the N-protected
amino epoxide include ethanol, methanol, isopropanol,
tetrahydrofuran, dioxane, dimethyl formamide and the like
(including mixtures thereof). Suitable bases for producing the
epoxide include alkali metal hydroxides, potassium t-butoxide, DBU
and-the like. A preferred base is potassium hydroxide.
[0208] Reaction of the N-protected amino epoxide or other suitably
activated intermediates with an amine is carried out neat, i.e. in
the absence of solvent, or in the presence of a polar solvent such
as lower alkanols, water, dimethylformamide or dimethylsulfoxide.
The reaction can be carried out conveniently between about
0.degree. C. and 120.degree. C., preferably between about
20.degree. C. and 100.degree. C. Alternatively, the reaction may be
carried out in the presence of an activating agent, such as
activated alumina in an inert solvent, preferably an ether, such as
diethyl ether, tetrahydrofuran, dioxane, or tert-butyl methyl
ether, conveniently from about room temperature to about
110.degree. C., as described by Posner and Rogers, J. Am Chem.
Soc., 99, p. 8208 (1977). Other activating reagents include lower
trialkylaluminum species, such as triethylaluminum, or
dialkylaluminum halide species, such as diethylaluminum chloride
(Overman and Flippin, Tetrahedron Letters, p. 195 (1981)).
Reactions involving these species are conveniently carried out in
inert solvents such as dichloromethane, 1,2-dichloroethane,
toluene, or acetonitrile between about 0.degree. C. and about
110.degree. C. Further methods of displacing leaving groups, or
opening epoxides with amines or their equivalents such as azides or
timethylsilyl cyanide (Gassman and Guggenheim, J. Am. Chem. Soc.
104, p. 5849 (1982)), are known and will be apparent to those of
ordinary skill in the art.
[0209] Compounds of formulae II and III, and
functionality-protected derivatives thereof, are useful as
intermediates for the preparation of compounds of formula I. In
those cases where L represents D', compounds of formula III may be
converted to compounds of formula I by reaction with
sulfonyl-activated species to form sulfonamides, sulfonyl ureas,
thiocarbamates and the like. Methods for preparing such
sulfonyl-activated species are well within the ordinary skill of
the art. Typically, sulfonyl halides are used to obtain
sulfonamides. Many sulfonyl halides are commercially available;
others may be easily obtained using conventional synthetic
techniques (Gilbert, E. E. "Recent Developments in Preparative
Sulfonation and Sulfation" Synthesis 1969: 3 (1969) and references
cited therein; Hoffman, R. V. "M-Trifluoromethylbenzenesulfonyl
Chloride" Org. Synth. Coll. Vol. VII, John Wiley and Sons (1990);
Hartman, G. D. et. al. "4-Substituted Thiophene-and
Furan-2-sulfonamides as Topical Carbonic Anhydrase Inhibitors" J.
Med. Chem., 35, p. 3822 (1992) and references cited therein.
Sulfonyl ureas are usually obtained by the reaction of an amine
with sulfuryl chloride or a suitable equivalent such as
sulfuryl-bis-imidazole or sulfuryl-bis-N-methyl imidazole.
Thiocarbamates are typically obtained by the reaction of an alcohol
with sulfuryl chloride or a suitable equivalent such as
sulfuryl-bis-imidazole or sulfuryl-bis-N-methyl imidazole.
[0210] In the case of compounds of formula III wherein L is
hydrogen, conversion of the resultant primary amine to a secondary
amine may be carried out by known techniques. Such techniques
include reaction with an alkyl halide of alkyl sulfonate, or by
reductive alkylation with an aldehyde or carboxylic acid or
activated derivative thereof using, for instance, catalytic
hydrogenation or sodium cyanoborohydride (Borch et al., J. Am.
Chem. Soc., 93, p. 2897 (1971)). Alternatively, the primary amine
may be acylated followed by reduction with borane or another
suitable reducing reagent, for example, as described by Cushman et
al., J. Org. Chem., 56, p. 4161 (1991). This technique is
especially useful in compounds of formula III where B is absent and
A represents a protecting group such as tert-butoxycarbonyl (Boc)
or benzyloxycarbonyl (Cbz).
[0211] If variable A of a particular compound of formula I
represents a removable protecting group, removal of that group
followed by reaction of the resulting amine with an appropriate
activated reagent will advantageously yield a different compound of
formula I. For instance, reaction with an activated carboxylate,
such as an acyl halide (e.g., acid fluo-rides, acid chlorides, and
acid bromides), an activated ester such as nitrophenyl ester or
1-hydroxysuccinimide (HOSU) ester, an anhydride such as the
symmetrical anhydride or isobutyl anhydride, or mixed
carbonic-phosphoric or carbonic-phosphinic anhydrides, will yield
the corresponding amide. Ureas may be obtained by reaction with
isocyanates or amines in the presence of bis-activated carbonic
acid derivatives such as phosgene or carbonyldiimdazole. Carbamates
may be obtained by reaction with chlorocarbonates, with carbonates
esterified with leaving groups such as 1-hydroxybenzotriazole
(HOBT) or HOSu, or with alcohols in the presence of bis-activated
carbonic acid derivatives such as phosgene or carbonyldiimdazole.
It will be readily recognized that in order to facilitate specific
reactions, the protection of one or more potentially reactive
groups followed by subsequent removal of that group may be
required. Such modification to the reaction schemes outlined above
are within the ordinary skill of the art.
[0212] If variable B of a particular compound of formula I is
absent and variable A of that compound represents a removable
protecting group, removal of A, followed by reaction of the
resulting amine with an amino acid or suitably N-protected
derivative thereof, followed by a subsequent reaction of the free
a-amine if present, as described above, will yield a further
compound of formula I. The addition of amino acids and their
derivatives is accomplished by well known methods of peptide
synthesis. Some of these methods are generally set forth in
Bodanszky and Bodanszky, "The Practice of Peptide Synthesis",
Springer-Verlag, Berlin, Germany (1984) and in the "The Peptides",
Gross and Meinhofer (Eds); Academic Press, 1979, Vols. I-III, which
are incorporated herein by reference.
[0213] Typically, for solution phase synthesis of peptides, the
a-amine of the amino acid to be coupled is protected by Boc, Cbz,
allyloxycarbonyl (Alloc) or 9-fluorenylmethoxycarbonyl (Fmoc),
while the free carboxyl is activated by reaction with a
carbodiimide such as dicyclohexylcarbodiimid- e (DCC),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC),
or diisopropylcarbodiimide (DIC), optionally in the presence of a
catalyst such as HOBT, HOSu, or dimethylaminopyridine (DMAP). Other
methods which proceed through the intermediacy of activated esters,
acid halides, enzyme-activated amino acids and anhydrides including
N-carboxy-anhydrides, symmetrical anhydrides, mixed carbonic
anhydrides, carbonic-phosphinic and carbonic-phosphoric anhydrides
are also suitable. After the peptide has been formed, protecting
groups may be removed by methods described in the references listed
above, such as by hydrogenation in the presence of a palladium,
platinum or rhodium catalyst, treatment with sodium in liquid
ammonia, hydrochloric, hydrofluoric, hydrobromic, formic,
trifluoromethanesulfonic, or trifluoroacetic acid, secondary
amines, fluoride ion, trimethylsilyl halides including bromide and
iodide, or alkali.
[0214] One particularly useful synthetic scheme for producing
sulfonamides of formula XV is shown below: 607
[0215] Compounds of formula X may be advantageously synthesized
from readily available starting materials (see D. P. Getman, J.
Med. Chem., 36, p. 288 (1993)). Each step of the above synthetic
scheme may be carried out as generally described above.
[0216] A particularly useful synthetic scheme for producing the
preferred sulfonamides of formula XXII is shown below: 608
[0217] Compounds of formula XX may be advantageously synthesized
from readily available starting materials (see B. E. Evans et al.,
J. Org. Chem., 50, p. 4615 (1985)). Each step of the above
synthetic scheme may be carried out as generally described
above.
[0218] After converting a compound of formula XX to a compound of
formula XXI, as detailed in the previous reaction scheme, the
compound of formula XXI may alternatively be reacted with an amino
acid or amino acid derivative, as described generally above, to
yield a preferred compound of formula XXXI. A particularly useful
synthetic scheme utilizing this strategy is set forth below:
609
[0219] As can be appreciated by the skilled artisan, the above
synthetic schemes are not intended to comprise a comprehensive list
of all means by which the compounds described and claimed in this
application may be synthesized. Further methods will be evident to
those of ordinary skill in the art.
[0220] The compounds of this invention may be modified by appending
appropriate functionalites 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.
[0221] The compounds of formula I are characterized by a superior
ability to inhibit HIV protease activity and viral replication. We
believe that this is due to specific steric and electronic
interactions between the protease and compounds of formula I. This
belief stems from our analysis of the structural basis for the
activity of compounds of formula I, in view of the known crystal
structures of HIV protease and bound inhibitors, such as the
structure reported in Miller et al. "Structure of Complex of
Synthetic HIV-1 Protease with a Substrate-Based Inhibitor at 2.3
.ANG. Resolution", Science, vol. 246, pp. 1149-1152 (1989), which
is incorporated herein by reference, as well as structures
determined in our laboratories. According to these structures, the
active site of HIV aspartyl protease is defined by a deep groove
containing subpockets for accommodation of various side chains of
the protease substrate--referred to as P.sub.1-P.sub.n and
P.sub.1'-P.sub.n', according to conventional protease nomenclature.
In the center of the groove, lie two aspartic acid residues (Asp25
and Asp25' according to the numbering system of Miller et al.) in a
manner typical of the active site aspartates of known aspartyl
proteases, which are believed to be the catalytic residues of the
enzyme. The groove is covered by two C.sub.2-symmetrically disposed
"flaps" which also make various direct and indirect contacts with
bound substrates.
[0222] We believe that the substituents A, D, D' and E of the
compounds of formula I associate with HIV protease by way of
hydrophobic forces in the binding pockets of the enzyme. We also
believe that the sulfonamide group hydrogen binds tightly to a
water molecule held by hydrogen bonds to the flaps of the protease
("the flap water molecule"; water molecule 511, according to the
Miller et al. numbering system).
[0223] In view of the above discovery, an alternative embodiment of
this invention relates to novel HIV protease inhibitors possessing
certain structural and physicochemical features. We have discovered
that compounds possessing the following novel combination of
features are surprisingly effective HIV protease inhibitors:
[0224] (1) a first and a second hydrogen bond acceptor moiety, at
least one of which is more highly polarizable than a carbonyl, said
moieties being the same or different, and being capable of hydrogen
bonding with the hydrogen atoms of the flap water molecule of an
HIV aspartyl protease when the compound is bound thereto;
[0225] (2) substantially hydrophobic moieties which associate with
the P.sub.1 and P.sub.1' binding pockets of said HIV aspartyl
protease when the compound is bound thereto;
[0226] (3) a third hydrogen bonding moiety, which may be either a
hydrogen bond donor or acceptor, capable of simultaneously hydrogen
bonding to Asp25 and Asp25' of said HIV aspartyl protease when the
compound is bound thereto;
[0227] (4) an additional occupied volume of space of at least 100
.ANG..sup.3 when the compound is bound to the active site of said
HIV aspartyl protease, said space overlapping with the volume of
space that would be filled by a native substrate of said HIV
aspartyl protease or a nonhyrolyzable isostere thereof;
[0228] (5) a deformation energy of binding of the compound to said
HIV aspartyl protease of not greater than 10 kcal/mole; and
[0229] (6) a neutral or favorable enthalpic contribution from the
sum of all electrostatic interactions between the compound and the
protease when the compound is bound to said HIV aspartyl
protease.
[0230] Compounds having the above-cited features can be readily
identified or designed by one of ordinary skill in the art using a
combination of chemical reasoning and computational methods. For
example, those of ordinary skill in the art can readily identify or
choose hydrogen bonding and hydrophobic moieties or groups required
in features (1)-(3), while features (4)-(6) can be ascertained
using well known computational methods for determination of
structural (e.g. conformational) and energetic properties of
molecules.
[0231] Furthermore, compounds characterized by features (1) through
(6) listed above may be obtained using any conventional technique,
including chemical synthesis and natural product isolation. We
prefer using the synthetic schemes detailed above for compounds of
formula I.
[0232] We have discovered that when an HIV protease inhibitor forms
hydrogen bonds to the flap water molecule through two hydrogen
bonding moieties, at least one of which is more highly polarizable
than a carbonyl, the ability of those compounds to inhibit HIV
protease activity is dramatically improved, as compared with
conventional HIV protease inhibitors.
[0233] While not wishing to be bound by theory, we believe that the
strong hydrogen bonds that form between the flap water molecule and
the two hydrogen bonding moieties, at least one of which is more
highly polarizable than a carbonyl, lower the overall binding
energy of the inhibitor. Most HIV protease inhibitors known in the
art utilize only carbonyl groups for hydrogen bonding to the flap
water molecule and are, thus, inferior to those of the present
invention. We believe that the increased polarization that results
from the large dipole moment of the highly polarizable hydrogen
bonding moiety (as compared to the dipole moment of a carbonyl
moiety) creates a stronger and tighter hydrogen bond with the flap
water molecule. We prefer to utilize tetravalent oxygenated sulfur,
hexavalent oxygenated sulfur and pentavalent oxygenated phosphorus
as the highly polarizable hydrogen bonding moiety. Tetravalent
oxygenated sulfur and hexavalent oxygenated sulfur are more
preferred as the highly polarizable hydrogen bonding moiety.
Hexavalent oxygenated sulfur (--SO.sub.2--) is most preferred.
[0234] We have found that when the highly polarizable hydrogen
bonding moiety is a sulfonamide, the overall binding energy of the
inhibitor is particularly low. We believe that this increased
stability is due to particular conformational characteristics of
the sulfonamide S--N bond. Specifically, the sulfonamide S--N bond
exists in only two low-energy rotamers (see J. B. Nicholas et al.,
J. Phys. Chem., 95, p. 9803 (1991) and R. D. Bindal et al., J. Am.
Chem. Soc., 112, p. 7861 (1990)). This has the effect of locking
that portion of the molecule into a favorable conformation wherein
one or both of the highly polarized S.dbd.O oxygens can be involved
in hydrogen bonding interactions with the flap water.
[0235] The remaining five structural and physicochemical features
recited above (i.e., features (2) through (6)) are generally
recognized in the art to improve the ability of a compound to
competitively inhibit HIV protease activity. Although there are
several other features thought to increase the inhibitory property
(such as binding of the inhibitor backbone to the enzyme), we have
discovered that the combination of the five above-cited elements
alone, together with novel element (1), typifies effective HIV
protease inhibitors.
[0236] In general, the binding energy of a particular protease
inhibitor is lowered when hydrophobic moieties on the inhibitor are
located so as to associate with the enzyme's hydrophobic binding
pockets. In the case of HIV-1 protease, the location and nature of
the P.sub.1 and P.sub.1' binding pockets are known to those of
ordinary skill in the art (see, for example, M. Miller et al.,
cited above). Substantially hydrophobic side chains which fit into
the well defined P.sub.1 and P.sub.1' binding pockets are also
known to those in the art. Preferred side chains are located within
4 .ANG. of the enzyme when bound to HIV protease. Preferred
hydrophobic side chains include those substantially similar to
those of hydrophobic natural and unnatural .alpha.-amino acids,
including alanine, valine, leucine, isoleucine, methionine,
phenylalanine, .alpha.-amino isobutyric acid, alloisoleucine,
tyrosine, and tryptophan. Insofar as a portion of this side chain
is in contact with bulk solvent or protrudes out of the enzyme, it
is not considered to be wholly within P.sub.1 or P.sub.1' and may
contain polar functionality such as a charged amine at that
location.
[0237] It has also been established in the art that the presence of
a hydroxyl group within hydrogen bond proximity to the two
catalytic aspartic acid residues of HIV protease (Asp25 and Asp25')
is an important feature of an effective HIV protease inhibitor
(see, for example, R. Bone et al., "X-ray Crystal Structure of the
HIV Protease Complex with L-700,417, an Inhibitor with Pseudo
C.sub.2 Symmetry", J. Am. Chem. Soc., 113, pp. 9382-84 (1991)). It
is further understood that the geometry of the Asp-binding hydrogen
bonding moiety is of particular importance. Although we prefer to
use a hydroxyl group at this position, any hydrogen bonding moiety
that is capable of forming hydrogen bonds with the Asp residues is
acceptable. Such hydrogen bonding moieties are known to those of
skill in the art (e.g., phosphinic acid (D. Grobelny et al.,
Biochem. Biophys. Res. Commun., 169, p. 1111 (1990)).
[0238] It is further understood that binding of competitive
inhibitors to HIV protease is optimally accomplished by having the
inhibitor traverse a volume overlapping that occupied by the native
polypeptide substrate when it is bound to the active site of the
enzyme. Effective HIV protease inhibitors typically have a
relatively small difference in energy between their bound and free
states (i.e., a small deformation energy of binding). The most
preferred HIV protease inhibitors of this invention have a
deformation energy of binding of not greater than 10 kcal/mole
(preferably, not greater than 7 kcal/mole). It should be noted,
however, that HIV protease inhibitors may interact with HIV
protease in more than one conformation which is similar in overall
binding energy (see K. H. M. Murthy, J. Biol. Chem., 267, (1992)).
In those cases, the deformation energy of binding is taken to be
the difference between the energy of the free compound and the
average energy of the conformations observed when the inhibitor
binds to the enzyme.
[0239] Furthermore, it is understood that the most effective
protease inhibitors also lack repulsive electrostatic interaction
with the target protease in their bound state. Such
non-complementary (e.g., electrostatic) interactions include
repulsive charge-charge, dipole-dipole and charge-dipole
interactions. Specifically, in the most preferred HIV protease
inhibitors of this invention, the sum of all electrostatic
interactions between the compound and the enzyme when the compound
is bound to HIV protease makes a neutral or favorable contribution
to the enthalpy of binding.
[0240] Preferred compounds characterized by the above features
(1)-(6) are compounds of formula XL:
Z.sup.1--Q.sup.1--L.sup.1--M--L.sup.2--Q.sup.2--Z.sup.2 (XL)
[0241] wherein:
[0242] Q.sup.1 and Q.sup.2 are independently hydrogen bond acceptor
moieties capable of binding with the hydrogen atoms of the flap
water molecule of an HIV aspartyl protease, with the proviso that
at least one of Q.sup.1 or Q.sup.2 is more highly polarizable than
a carbonyl;
[0243] M is a hydrogen bonding moiety, which may be either a
hydrogen bond donor or acceptor, capable of simultaneously hydrogen
bonding to Asp25 and Asp25' of said HIV aspartyl protease;
[0244] L.sup.1 and L.sup.2 are independently acyclic or cyclic
linker moieties; and
[0245] each of Z.sup.1 and Z.sup.2 may be optionally present and,
if present, are independently selected from groups which occupy a
volume of space overlapping with the volume of space that would be
filled by the native substrate of said HIV aspartyl protease.
[0246] More preferred compounds of formula XL contain at least one
group Q.sup.1 or Q.sup.2 comprising --SO.sub.2--. Most preferrred
compounds of formula XL contain at least one group Q.sup.1 or
Q.sup.2 comprising a substituted sulfonamide.
[0247] In one embodiment of this invention, compounds of formula XL
may be further constrained by "conformational locks", such as a
macrocyclic ring structure. Such constraints are well known in the
art of peptidomimetics and may result in compounds with strong
biological activity. See, for example, Dhanoa, D. S. et al. "The
Synthesis of Potent Macrocyclic Renin Inhibitors" Tetrahedron Lett.
33, 1725 (1992) and Flynn, G. A. et al. "An Acyl-Iminium Ion
Cyclization Route to a Novel Conformationally Restricted Dipeptide
Mimic: Applications to Angiotensin-Converting Enzyme Inhibition"
J.Am. Chem. Soc. 109, 7914 (1989)).
[0248] This invention also includes novel methods for accurate
identification, design, or prediction of HIV inhibitors
characterized by structural and physicochemical features (1)
through (6). By virtue of these methods, the skilled artisan can
routinely predict and produce particularly effective HIV protease
inhibitors.
[0249] We have found that the following method for identification,
design or prediction of effective HIV protease inhibitors is
particularly useful;
[0250] (a) selecting a candidate compound of defined chemical
structure containing a first and a second hydrogen bond acceptor
moiety, at least one of which is more highly polarizable than a
carbonyl, said moieties being the same or different; a third
hydrogen bonding moiety, which may be either a hydrogen bond donor
or acceptor; and at least two substantially hydrophobic
moieties;
[0251] (b) determining a low-energy conformation for binding of
said compound to the active site of an HIV aspartyl protease;
[0252] (c) evaluating the capability of said first and second
hydrogen bond acceptor moieties to form hydrogen bonds to the flap
water molecule of said HIV aspartyl protease when said compound is
bound thereto in said conformation;
[0253] (d) evaluating the capability of said substantially
hydrophobic moieties to associate with the P.sub.1 and P.sub.1'
binding pockets of said HIV aspartyl protease when said compound is
bound thereto in said conformation;
[0254] (e) evaluating the capability of said third hydrogen bonding
moiety to form hydrogen bonds to Asp25 and Asp25' of said HIV
aspartyl protease when said compound is bound thereto in said
conformation;
[0255] (f) evaluating the overlap of the occupied volume of said
compound when said compound is bound to said HIV aspartyl protease
in said conformation and the occupied volume of a native substrate
of HIV aspartyl protease or a nonhydrolyzable isostere thereof,
when said polypeptide is bound to said HIV aspartyl protease;
[0256] (g) evaluating the deformation energy of binding of said
compound to said HIV aspartyl protease;
[0257] (h) evaluating the enthalpic contribution of the sum of all
electrostatic interactions between said compound and said HIV
aspartyl protease when said compound is bound thereto in said
conformation; and
[0258] (i) accepting or rejecting said candidate compound as an HIV
protease inhbitor based upon the determinations and evaluations
carried out in steps (b) through (h).
[0259] Using the novel combination of steps set forth in this
screening method, the skilled artisan can advantageously avoid time
consuming and expensive experimentation to determine enzymatic
inhibition activity of particular compounds. The method is also
useful for facilitating rational design of HIV protease inhibitors
and anti-HIV viral agents, including therapeutic and prophylactic
agents against HIV infection. Accordingly, the present invention
relates to such inhibitors and anti-viral agents produced by the
screening method described above.
[0260] A variety of conventional techniques may be used to carry
out each of the above evaluations. Generally, these techniques
involve determining the location and binding proximity of a given
moiety, the occupied volume of space of a bound compound, the
deformation energy of binding of a given compound and electrostatic
interaction energies. Examples of conventional techniques useful in
the above evaluations include: quantum mechanics, molecular
mechanics, molecular dynamics, Monte Carlo sampling, systematic
searches and distance geometry methods (G. R. Marshall, Ann. Ref.
Pharmacol. Toxicol., 27, p. 193 (1987)). Specific computer software
has been developed for use in carrying out these methods. Examples
of programs designed for such uses include: Gaussian 92, revision C
(M. J. Frisch, Gaussian, Inc., Pittsburgh, Pa. .COPYRGT.1992);
AMBER, version 3.0 (U.C. Singh, University of California at San
Francisco, .COPYRGT.1992); QUANTA/CHARMM (Molecular Simulations,
Inc., Burlington, Mass. .COPYRGT.1992); and Insight II/Discover
(Biosysm Technologies Inc., San Diego, Calif. .COPYRGT.1992). These
programs may be implemented, for instance, using a Silicon Graphics
workstation, IRIS 4D/35 or IBM RISC/6000 workstation model 550.
Other hardware systems and software packages will be known and of
evident applicability to those skilled in the art.
[0261] Additional analysis of the actual detailed interactions of
the HIV protease-inhibitor complex can be employed to ascertain
more specifically the binding associations between the enzyme and
the bound inhibitor. Such analysis may be carried out, for example,
by studying a solution of the complex by single- and
multi-dimensional NMR techniques. Advantageously, the enzyme and/or
the inhibitor may be enriched with stable isotopes such as
.sup.13C, .sup.15N and .sup.2H to more easily determine binding
conformation and proximity. Techniques, such as isotope editing,
may be used to enhance the resolution with which the interactions
are observed.
[0262] Either as an alternative or a supplemental analysis, the HIV
protease-inhibitor complex may be studied by single crystal X-ray
diffraction. The process of determining the structures of
protein/inhibitor complexes using the X-ray techniques described
above is well known and has been used for many different complexes
(see T. L. Blundel and L. N. Johnson, Protein Crystallography,
Academic Press, (1976) and Methods in Enzymology, volumes 114 and
115, H. W. Wyckoff et al., eds., Academic Press (1985)). This
technique can employ, for instance, a highly purified preparation
of HIV protease complexed with an inhibitor of interest in a
buffered solution (typically at a pH of between about 4.5 and about
8.0). The complex is allowed to crystallize in the presence of a
precipitation agent (such as ammonium sulfate) under conditions
which yield single crystals of the complex. Specific conditions for
crystallizing HIV protease with various inhibitors have been well
documented (see, for example, G. B. Dreyer et al., Biochemistry,
31, p. 6646 (1992)). Application of a concentrated X-ray beam to an
appropriately prepared and mounted crystal (preferably, an X-ray
beam from a rotating anode X-ray generator or synchrotron) will
yield a diffraction pattern from the reflected X-ray beam.
[0263] Detection of the diffracted rays may be carried out by
visualizing photographic paper exposed to the diffracted X-rays or
alternatively, by using a multiwire area detector (such as that
manufactured by Siemens Analytical X-Ray Instruments, Inc.
(Madison, Wis.)) or an R-axis II image plate system from Rigaku
Corporation (distributed by Molecular Structure Corporation, The
Woodlands, Tex.). Other systems for generating and collecting X-ray
diffraction data will be known to those of ordinary skill in the
art.
[0264] Refinement of the X-ray diffraction data yields a three
dimensional structure. Computer software (such as X-PLOR (Yale
University, .COPYRGT.1992, distributed by Molecular Simulations,
Inc.) has been developed to carry out this refinement.
[0265] In general, using the above techniques with an appropriately
prepared crystalline complex, a structure may be refined to about
2-3 .ANG. with an R value of about 0.25 or less. As the skilled
artisan can appreciate, these values are adequate to determine the
interactions between HIV protease and a given compound such that it
will be clear if features (1) through (6) are present and
consequently, whether that given compound is an HIV aspartyl
protease inhibitor. Thus, additional inhibitors according to this
invention may be designed and predicted based on a combination of
crystallographic structural information and computational
analysis.
[0266] For example, to predict the binding of a candidate inhibitor
according to this invention, the inhibitor is examined to determine
whether the molecule contains functionality which is not well
represented by the existing forcefield models in CHARMM (Molecular
Simulations Incorporated, Burlington, Mass.) or AMBER (Professor P.
A. Kollman, UCSF). If any functionality is not well represented, we
then examine all published structural information for molecules
containing such functionality, and in some cases perform high-level
ab initio calculations on simple molecules containing these
functionalities to determine their preferred conformations and the
energy differences between various conformations. More accurate
parameters describing these functional groups may then be derived
for the CHARMM and/or AMBER forcefields and used in subsequent
calculations.
[0267] Next, the candidate inhibitor is aligned in 3-dimensional
space with other, related inhibitors whose bound conformations have
previously been determined by x-ray crystallography. Both Van der
Walls volume and electrostatic potentials are used to direct the
alignment process. The alignment is typically done with software
like Quanta (Molecular Simulations) or InsightII (Biosym
Technologies, San Diego, Calif.). This alignment can be done
manually within this software, or more automated alignment
procedures within the software (e.g. the "superimpose" option of
Quanta or the "APEX" module of InsightII) may be used. The result
of this alignment is a first guess of the "bound" conformation of
the candidate inhibitor. This inhibitor is then docked in the
active site of HIV protease, and the confomation is energy
minimized with the enzyme atoms held fixed in space. These
minimizations are typically done using the CHARMM or AMBER
forcefields.
[0268] Because inhibitors can sometimes bind in multiple or
unexpected conformations within an active site, we often then carry
out further searches of the bound conformation of the
enzyme-inhibitor complex. For example, a variety of Monte Carlo
search techniques (e.g. as found in the Conformational Search
Module of Quanta) may be used, along with high-temperature dynamics
and simulated annealing. These search techniques reveal whether
there are alternative, reasonable low-energy confomrations in which
the inhibitor may bind to the enzyme. The effects of salvation and
desolation in the formation of the various enzyme-inhibitor
complexes may be estimated with programs such as DELPHI (Biosym),
Polaris (Molecular Simulations) and AMSOL (Professor C. Cramer,
University of Minnesota). The result of this searching is a set of
one or more bound conformations for the candidate inhibitor.
[0269] For each of the low-energy conformations, waters may then be
added to the active site of the enzyme and the entire system
relaxed. Finally, molecular dynamics simulations may be used to
study the detailed motions of the enzyme, the inhibitor, and
related water molecules.
[0270] The final set of remaining low-energy conformations
(typically a very small number) represents our predictions of the
bound conformation of the candidate inhibitor. Each conformation
includes our estimate of the dynamic flexibility of the entire
system (inhibitor, enzyme, and waters).
[0271] The more advanced methodology is typically applied to the
study of the first few compounds in a series, when there are the
greatest uncertainties about the possible binding mode(s) in the
enzyme active site. For later compounds within a series, the low
energy conformers obtained from the searches on earlier compounds
provide information about the possible low energy conformers of the
inhibitor compounds. In addition, crystallographic information
about the conformation of the bound complexes of earlier compounds
within a series is often available. This prior computational and
structural work advantageously facilitates the prediction of the
bound conformation of candidate inhibitor molecules.
[0272] To exemplify the above screening method, we have carried out
the following evaluation of compound 140 (Table II), a preferred
compound of this invention, as described below.
[0273] Prediction of Binding Conformation and Energy of Compound
140 to HIV Protease
[0274] The forcefield for the benzenesulfonamide portion of
compound 140 was derived from ab initio calculations and
incorporated into the AMBER forcefield. The latest CHARMM
forcefield parameters for this moiety were found to be adequate for
energy minimization studies and are used in all Quanta/CHARMM
calculations.
[0275] The low energy conformers obtained from the conformational
searches on earlier compounds in the sulfonamide series (such as
compound 16) provided information about the possible low energy
conformers of compound 140. These low energy conformers were
aligned in 3-dimensional space with other related inhibitors whose
bound conformations have previously been determined by x-ray
crystallography. This alignment process was carried out manually
within Quanta and, in some cases, was assisted with the
"conformational search" option of Quanta. The reference crystal
structure used in this alignment was the complex of HIV-1 protease
with compound 16. This inhibitor structure was energy minimized in
the active site of the enzyme using Quanta/CHARM. The enzyme atoms
were held fixed during this minimization. Only the flap water was
included. Later simulations allowed the enzyme to relax and used a
variety of dielectric approximations. A single low-energy
conformation which was consistent with all previous conformational
simulations and crystallographic data was obtained (see FIG. 1).
This predicted binding conformation was later found to be
essentially in agreement with the results obtained by x-ray
crystallography (see FIGS. 2 and 3).
[0276] As discussed above, the novel compounds of the present
invention are excellent ligands for aspartyl proteases,
particularly HIV-1 and HIV-2 proteases. Accordingly, these
compounds are capable of targeting and inhibiting late stage events
in HIV replication, i.e., the processing of the viral polyproteins
by HIV encoded proteases. Such compounds inhibit the proteolytic
processing of viral polyprotein precursors by inhibiting aspartyl
protease. Because aspartyl protease is essential for the production
of mature virions, inhibition of that processing effectively blocks
the spread of virus by inhibiting the production of infectious
virions, particularly from chronically infected cells. Compounds
according to this invention advantageously inhibit the ability of
the HIV-1 virus to infect immortalized human T cells over a period
of days, as determined by an assay of extracellular p24 antigen--a
specific marker of viral replication. Other anti-viral assays have
confirmed the potency of these compounds.
[0277] The compounds of this invention may be employed in a
conventional manner for the treatment of viruses, such as HIV and
HTLV, which depend on aspartyl proteases for obligatory events in
their life cycle. Such methods of treatment, their dosage levels
and requirements may be selected by those of ordinary skill in the
art from available methods and techniques. For example, a compound
of this invention may be combined with a pharmaceutically
acceptable adjuvant for administration to a virally-infected
patient in a pharmaceutically acceptable manner and in an amount
effective to lessen the severity of the viral infection.
[0278] Alternatively, the compounds of this invention may be used
in vaccines and methods for protecting individuals against viral
infection over an extended period of time. The compounds may be
employed in such vaccines either alone or together with other
compounds of this invention in a manner consistent with the
conventional utilization of protease inhibitors in vaccines. For
example, a compound of this invention may be combined with
pharmaceutically acceptable adjuvants conventionally employed in
vaccines and administered in prophylactically effective amounts to
protect individuals over an extended period time against HIV
infection. As such, the novel protease inhibitors of this invention
can be administered as agents for treating or preventing HIV
infection in a mammal.
[0279] The compounds of formula I, especially those having a
molecular weight of less than about 700 g/mole, may be readily
absorbed by the bloodstream of mammals upon oral administration.
Compounds of formula I having a molecular weight of less than about
600 g/mole are most likely to demonstrate oral availability. This
surprisingly impressive oral availability makes such compounds
excellent agents for orally-administered treatment and prevention
regimens against HIV infection.
[0280] The compounds of this invention may be administered to a
healthy or HIV-infected patient either as a single agent or in
combination with other anti-viral agents which interfere with the
replication cycle of HIV. By administering the compounds of this
invention with other anti-viral agents which target different
events in the viral life cycle, the therapeutic effect of these
compounds is potentiated. For instance, the co-administered
anti-viral agent can be one which targets early events in the life
cycle of the virus, such as cell entry, reverse transcription and
viral DNA integration into cellular DNA. Anti-HIV agents targeting
such early life cycle events include, didanosine (ddI), alcitabine
(ddC), d4T, zidovudine (AZT), polysulfated polysaccharides, sT4
(soluble CD4), ganiclovir, dideoxycytidine, trisodium
phosphonoformate, eflornithine, ribavirin, acyclovir, alpha
interferon and trimenotrexate. Additionally, non-nucleoside
inhibitors of reverse transcriptase, such as TIBO or nevirapine,
may be used to potentiate the effect of the compounds of this
invention, as may viral uncoating inhibitors, inhibitors of
trans-activating proteins such as tat or rev, or inhibitors of the
viral integrase.
[0281] Combination therapies according to this invention exert a
synergistic effect in inhibiting HIV replication because each
component agent of the combination acts on a different site of HIV
replication. The use of such combinations also advantageously
reduces the dosage of a given conventional anti-retroviral agent
which would be required for a desired therapeutic or prophylactic
effect as compared to when that agent is administered as a
monotherapy. These combinations may reduce or eliminate the side
effects of conventional single anti-retroviral agent therapies
while not interfering with the anti-retroviral activity of those
agents. These combinations reduce potential of resistance to single
agent therapies, while minimizing any associated toxicity. These
combinations may also increase the efficacy of the conventional
agent without increasing the associated toxicity. In particular, we
have discovered that these compounds act synergistically in
preventing the replication of HIV in human T cells.
[0282] Preferred combination therapies include the administration
of a compound of this invention with AZT, ddI, ddC or d4T.
[0283] Alternatively, the compounds of this invention may also be
co-administered with other HIV protease inhibitors such as Ro
31-8959 (Roche), L-735,524 (Merck), XM 323 (Du-Pont Merck) and
A-80,987 (Abbott) to increase the effect of therapy or prophylaxis
against various viral mutants or members of other HIV quasi
species.
[0284] We prefer administering the compounds of this invention as
single agents or in combination with retroviral reverse
transcriptase inhibitors, such as derivatives of AZT, or other HIV
aspartyl protease inhibitors. We believe that the co-administration
of the compounds of this invention with retroviral reverse
transcriptase inhibitors or HIV aspartyl protease inhibitors may
exert a substantial synergistic effect, thereby preventing,
substantially reducing, or completely eliminating viral infectivity
and its associated symptoms.
[0285] The compounds of this invention can also be administered in
combination with immunomodulators (e.g., bropirimine, anti-human
alpha interferon antibody, IL-2, GM-CSF, methionine enkephalin,
interferon alpha, diethyldithiocarbamate, tumor necrosis factor,
naltrexone and rEPO); and antibiotics (e.g., pentamidine
isethiorate) to prevent or combat infection and disease associated
with HIV infections, such as AIDS and ARC.
[0286] When the compounds of this invention are administered in
combination therapies with other agents, they may be administered
sequentially or concurrently to the patient. Alternatively,
pharmaceutical or prophylactic compositions according to this
invention may be comprised of a combination of an aspartyl protease
inhibitor of this invention and another therapeutic or prophylactic
agent.
[0287] Although this invention focuses on the use of the compounds
disclosed herein for preventing and treating HIV infection, the
compounds of this invention can also be used as inhibitory agents
for other viruses which depend on similar aspartyl proteases for
obligatory events in their life cycle. These viruses include, as
well as other AIDS-like diseases caused by retroviruses, such as
simian immunodeficiency viruses, but are not limited to, HTLV-I and
HTLV-II. In addition, the compounds of this invention may also be
used to inhibit other aspartyl proteases, and in particular, other
human aspartyl proteases, including renin and aspartyl proteases
that process endothelin precursors.
[0288] Pharmaceutical compositions of this invention comprise any
of the compounds of the present invention, and pharmaceutically
acceptable salts thereof, with any pharmaceutically acceptable
carrier, adjuvant or vehicle. Pharmaceutically acceptable carriers,
adjuvants and vehicles that may be used in the pharmaceutical
compositions of this invention include, but are not limited to, ion
exchangers, alumina, aluminum stearate, lecithin, serum proteins,
such as human serum albumin, buffer substances such as phosphates,
glycine, sorbic acid, potassium sorbate, partial glyceride mixtures
of saturated vegetable fatty acids, water, salts or electrolytes,
such as protamine sulfate, disodium hydrogen phosphate, potassium
hydrogen phosphate, sodium chloride, zinc salts, colloidal silica,
magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based
substances, polyethylene glycol, sodium carboxymethylcellulose,
polyacrylates, waxes, polyethylene-polyoxypropyle- ne-block
polymers, polyethylene glycol and wool fat.
[0289] The pharmaceutical compositions of this invention may be
administered orally, parenterally, by inhalation spray, topically,
rectally, nasally, buccally, vaginally or via an implanted
reservoir. We prefer oral administration or administration by
injection. The pharmaceutical compositions of this invention may
contain any conventional non-toxic pharmaceutically-acceptable
carriers, adjuvants or vehicles. The term parenteral as used herein
includes subcutaneous, intracutaneous, intravenous, intramuscular,
intra-articular, intrasynovial, intrasternal, intrathecal,
intralesional and intracranial injection or infusion
techniques.
[0290] The pharmaceutical compositions may be in the form of a
sterile injectable preparation, for example, as a sterile
injectable aqueous or oleaginous suspension. This suspension may be
formulated according to techniques known in the art using suitable
dispersing or wetting agents (such as, for example, Tween 80) 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 mannitol, 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 diglycerides. 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 Ph. Helv or a similar alcohol.
[0291] The pharmaceutical compositions of this invention may be
orally administered in any orally acceptable dosage form including,
but not limited to, capsules, tablets, and aqueous suspensions and
solutions. In the case of tablets for oral use, carriers which are
commonly used include lactose and corn starch. Lubricating agents,
such as magnesium stearate, are also typically added. For oral
administration in a capsule form, useful diluents include lactose
and dried corn starch. When aqueous suspensions are administered
orally, the active ingredient is combined with emulsifying and
suspending agents. If desired, certain sweetening and/or flavoring
and/or coloring agents may be added.
[0292] The pharmaceutical compositions of this invention may also
be administered in the form of suppositories for rectal
administration. These compositions can be prepared by mixing a
compound of this invention with a suitable non-irritating excipient
which is solid at room temperature but liquid at the rectal
temperature and therefore will melt in the rectum to release the
active components. Such materials include, but are not limited to,
cocoa butter, beeswax and polyethylene glycols.
[0293] Topical administration of the pharmaceutical compositions of
this invention is especially useful when the desired treatment
involves areas or organs readily accessible by topical application.
For application topically to the skin, the pharmaceutical
composition should be formulated with a suitable ointment
containing the active components suspended or dissolved in a
carrier. Carriers for topical administration of the compounds of
this invention include, but are not limited to, mineral oil, liquid
petroleum, white petroleum, propylene glycol, polyoxyethylene
polyoxypropylene compound, emulsifying wax and water.
Alternatively, the pharmaceutical composition can be formulated
with a suitable lotion or cream containing the active compound
suspended or dissolved in a carrier. 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. The pharmaceutical compositions of this
invention may also be topically applied to the lower intestinal
tract by rectal suppository formulation or in a suitable enema
formulation. Topically-transdermal patches are also included in
this invention.
[0294] The pharmaceutical compositions of this invention may 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 solubilizing or dispersing agents known in the
art.
[0295] Dosage levels of between about 0.01 and about 100 mg/kg body
weight per day, preferably 0.5 and about 50 mg/kg body weight per
day of the active ingredient compound are useful in the prevention
and treatment of viral infection, including HIV infection.
Typically, the pharmaceutical compositions of this invention will
be administered from about 1 to about 5 times per day or
alternatively, as a continuous infusion. Such administration can be
used as a chronic or acute therapy. The amount of active ingredient
that may be combined with the carrier materials to produce a single
dosage form will vary depending upon the host treated and the
particular mode of administration. A typical preparation will
contain from about 5% to about 95% active compound (w/w).
Preferably, such preparations contain from about 20% to about 80%
active compound.
[0296] 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.
[0297] As the skilled artisan will appreciate, lower or higher
doses than those recited above may be required. Specific dosage and
treatment regimens for any particular patient will depend upon a
variety of factors, including the activity of the specific compound
employed, the age, body weight, general health status, sex, diet,
time of administration, rate of excretion, drug combination, the
severity and course of the infection, the patient's disposition to
the infection and the judgment of the treating physician.
[0298] The compounds of this invention are also useful as
commercial reagents which effectively bind to aspartyl proteases,
particularly HIV aspartyl protease. As commercial reagents, the
compounds of this invention, and their derivatives, may be used to
block proteolysis of a target peptide or may be derivatized to bind
to a stable resin as a tethered substrate for affinity
chromatography applications. These and other uses which
characterize commercial aspartyl protease inhibitors will be
evident to those of ordinary skill in the art.
[0299] In order that this invention be more fully understood, the
following 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.
[0300] General Materials and Methods
[0301] All temperatures are recorded in degrees Celsius. Thin layer
chromatography (TLC) was carried out using 0.25 mm thick E. Merck
silica gel 60 F.sub.254 plates and elution with the indicated
solvent system. Detection of the compounds was carried out by
treating the plate with an appropriate visualizing agent, such as
10% solution of phosphomolybdic acid in ethanol or a 0.1% solution
of ninhydrin in ethanol, followed by heating, and/or by exposure to
UV light or iodine vapors when appropriate. Thick layer silica gel
chromatography was also carried out using E. Merck 60 F.sub.254
plates ("prep plates") of 0.5, 1.0, or 2.0 mm thickness. Following
development of the plate, the band of silica containing the desired
compound was isolated and eluted with an appropriate solvent.
Analytical HPLC was carried out using a Water's Delta Pak, 5 .mu.M
silica, C18 reversed-phase column, 3.9 mm ID.times.15 cm L with a
flow rate of 1.5 mL/min using the following table:
8 Mobile phase: A = 0.1% CF.sub.3CO.sub.2H in H.sub.2O B = 0.1%
CF.sub.3CO.sub.2H in CH.sub.3CN Gradient: T = 0 min., A (95%), B
(5%) T = 20 min., A (0%), B (100%) T = 22.5 min., A (0%), B
(100%)
[0302] Preparative HPLC was also carried out using C.sub.18
reversed-phase media. HPLC retention times were recorded in
minutes. NMR spectral data was recorded using a Bruker AMX500,
equipped with either a reverse or QNP probe, at 500 MHz, and was
taken in the indicated solvent.
[0303] We have measured the inhibition constants of each compound
against HIV-1 protease using the method described essentially by M.
W. Pennington et al., Peptides 1990, Gimet, E. and D. Andrew, Eds.,
Escom; Leiden, Netherlands (1990).
[0304] Compounds of formula I were tested for their antiviral
potency in several virological assays. In the first assay, the
compounds were added as a solution in dimethylsulfoxide (DMSO) to a
test cell culture of CCRM-CEM cells, a strain of CD4+ human T-cell
lymphoma cells, previously acutely infected with HIV.sub.IIIb using
standard protocols (see Meek, T. D. et al., "Inhibition of HIV-1
protease in infected T-lymphocytes by synthetic peptide analogues",
Nature, 343, p. 90 (1990). Preferred compounds are those which are
able to inhibit 90% of viral infectivity at a concentration of 1
.mu.M or less. More preferred compounds are those which are able to
inhibit 90% of viral infectivity at a concentration of 100 nM or
less.
[0305] The effect of the compounds on inhibiting the replication of
the virus was measured by determining the HIV extracellular p24
antigen concentration using a commercial enzyme immunoassay
(obtained from Coulter Corporation, Hialeah, Fla.).
[0306] Depending on the cell type and the desired readout, syncytia
formation, reverse-transcriptase (RT) activity, or cytopathic
effect as assayed by a dye uptake method may also be used as
readouts of antiviral activity. See H. Mitsuya and S. Broder,
"Inhibition of the in vitro infectivity and cytopathic effect of
human T-lymphotropic virus type III/lymphoadenopathy-associated
virus (HTLV-III/LAV) by 2',3'-dideoxynucleosides", Proc. Natl.
Acad. Sci. USA, vol. 83, pp. 1911-1915 (1986). The effect of
compounds of formula I on clinical isolates of other HIV-1 strains
was determined by obtaining low-passaged virus from HIV-infected
patients and assaying the effect of the inhibitors in preventing
infection of the HIV virus in freshly prepared human peripheral
blood mononuclear cells (PBMCs).
[0307] Insofar as compounds of formula I are able to inhibit the
replication of the HIV virus in human T-cells and furthermore, may
be delivered orally to mammals, they are of evident clinical
utility for the treatment of HIV infection. These tests are
predictive of the compounds ability to inhibit HIV protease in
vivo.
EXAMPLE 1
[0308] A. Compound XI ((syn)-OH, D'=Benzyl).
[0309] 184 g of Brockman Super I grade neutral alumina was slurried
in sufficient diethyl ether to form a thick, stirrable suspension
and was treated with 7.48 mL of benzylamine. After stirring for 5
min, 7.28 g of
(1S,2S)-1-(N-benzyoxycarbonyl)-amino-2-phenylethyl-oxirane was
added and the mixture stirred for 15 h. The mixture was treated
with 15.28 g of di-tert-butylpyrocarbonate and 4.70 mL of
diisopropylethylamine. This mixture was stirred for 3.5 h, then
treated with 600 mL of methanol, allowed to stand for 3.5 h, and
filtered to yield a yellow oil, which was purified by silica gel
chromatography using a gradient of 0.5 to 1.5% methanol in
methylene chloride to yield 3.88 g of the desired product as a
white solid. Further washing the filter cake with methanol and with
3% ammonium hydroxide in methanol yielded 2.2 g of
4-benzylamino-2-N-benzylo- xycarbonylamino-3-hydroxy-1-phenylbutane
in several portions. Each of these portions was treated separately,
as a solution in methylene chloride, with 1.1 molar equivalents
each of di-tert butylpyrocarbonate and diisopropylethylamine,
followed by aqueous workup with water, 10% aqueous KHSO.sub.4, and
brine, drying over MgSO.sub.4, and concentration in vacuo. The
combined products of these reactions were purified by silica gel
chromatography using a gradient of 5% to 15% diethyl ether in
methylene chloride. The resulting pure fractions were collected and
combined with the previously purified product to yield 5.49 g of a
white solid. TLC: Rf=0.56, 5% methanol/CH.sub.2Cl.sub.2;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
[0310] B. Compound XII ((syn)-OH, D'=Benzyl).
[0311] A solution of 5.49 g of the resultant compound of Example 1A
in 40 mL of ethanol was hydrogenated under a slight positive
pressure of hydrogen in the presence of 380 mg of 10% palladium on
carbon for 16 h. After filtering and concentrating in vacuo, the
desired product was obtained as 4.03 g of a white solid. TLC:
Rf=0.21, 95:5:0.5 CH.sub.2Cl.sub.2/methanol/concentrated
NH.sub.4OH.
[0312] C. Compound XIII ((syn)-OH, A=Benzyloxycarbonyl,
D'=Benzyl).
[0313] A solution of 3.02 g of the resultant compound of Example 1B
in 150 mL of methylene chloride was treated with 4.35 g of
N.sup..alpha.-Cbz-N.sup..delta.-trityl asparagine, 1.16 g of
hydroxybenzotriazole hydrate, and 1.64 g of
1-(3-dimethylaminopropyl)-3-e- thylcarbodiimide hydrochloride. The
mixture was stirred for 16 h, then diluted with 3 volumes of
diethyl ether and washed sequentially with water, saturated
NaHCO.sub.3 solution, 10% KHSO.sub.4 solution, and brine. After
drying over MgSO.sub.4 and concentrating in vacuo, a yellow oil was
obtained which was purified by chromatography on a Florisil column
using a gradient of 0% to 25% EtOAc in CH.sub.2Cl.sub.2 as eluant
to yield 8.00 g of the title compound as a white foam. TLC:
Rf=0.51, 5% methanol/CH.sub.2Cl.sub.2; (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
[0314] D. Compound XIV ((syn)-OH, A=H, D'=Benzyl).
[0315] A solution of 7.90 g of the resultant compound of Example 1C
in 150 mL of ethanol was hydrogenated under a slight positive
pressure of hydrogen int he presence of 550 mg of 10% palladium on
carbon for 2.5 h, then ca. 50 mg more 10% palladium on carbon was
added, the mixture was then filtered and concentrated in vacuo to
give the desired product as 6.66 g of a white solid which was used
without subsequent purification. TLC: Rf=0.26, 95:5:0.5
CH.sub.2Cl.sub.2/methanol/concentrated NH.sub.4OH.
[0316] E. Compound XIV ((syn)-OH, A=Quinoline-2-Carbonyl,
D=Benzyl).
[0317] A suspension of 1.51 g of quinaldic acid and 6.17 g of the
resultant compound of Example 1D in 150 mL of acetonitrile was
treated with 1.52 mL of diisopropylethylamine and 3.58 g of BOP
reagent. The mixture was stirred for 14 h, then concentrated in
vacuo. The gummy residue was partitioned between ether and water,
and the organic layer was washed sequentially with brine, saturated
NaHCO.sub.3 solution, water, 10% KHSO.sub.4 solution, and brine,
then dried over MgSO.sub.4 and concentrated in vacuo. Subsequent
purification by silica gel chromatography using 0 to 8.5% solvent A
in methylene chloride (where solvent A is defined as 90:10:1,
methylene chloride/methanol/concentrated ammonium hydroxide)
yielded 5.79 g of the title compound as a white foam, along with
ca. 600 mg of slightly impure side fractions. TLC: Rf=0.41, 5%
methanol/CH.sub.2Cl.sub.2; (.sup.1H)-NMR (CDCl.sub.3) consistent
with structure.
[0318] F. Compound 1.
[0319] A 58 mg portion of the resultant compound of Example 1E was
treated with 1 mL of 90% aqueous TFA and allowed to stand for 17 h.
The mixture was concentrated in vacuo and the residue taken up in 3
mL of CH.sub.2Cl.sub.2, treated with 100 .mu.L of DIEA, and cooled
to 0.degree. C. To this solution was added 26 .mu.L of
benzenesulfonyl chloride, and the mixture was stirred for 18 h,
warming slowly to ambient temperature. After concentration of the
mixture in vacuo, the residue was purified by thick layer silica
gel chromatography using 5% MeOH/CH.sub.2Cl.sub.2 as eluant
followed by preparative reversed-phase C.sub.18HPLC using a linear
gradient of 40% to 100% CH.sub.3CN/H.sub.2O with 0.1% TFA for
elution to obtain 8.3 mg of the title compound. TLC: Rf=0.50, 5%
MeOH/CH.sub.2Cl.sub.2, HPLC: Rt=17.8 min. NMR (DMSO-d.sub.6)
.delta. 2.62 (dd, 1H); 2.76 9d, 2H); 2.80 (dd, 1H); 3.11, (d, 2H);
3.34 (dd, 1H); 4.59 (br s, 1H); 4.68 (br s, 1H); 3.97 (m, 1H); 4.20
(d, 1H), 4.35 (d, 1H); 4.68 (dd, 1H); 6.39 (d, 1H); 6.74 (t, 1H);
6.81 (t, 2H); 6.93 (d, 2H); 7.12-7.24 (m, 6H); 7.51 (t, 2H); 7.57
(t, 1H); 7.62 (dd, 1H); 7.77 (t, 2H), 7.96 (d, 1H); 8.09 (d, 1H);
8.16 (d, 1H); 8.31 (d, 1H); 8.53 (d, 1H).
EXAMPLE 2
[0320] Compound 2.
[0321] A 150 mg portion of the resultant compound of Example 1E was
dissolved in 1 mL of 90% aqueous TFA and stirred at ambient
temperature overnight, then concentrated in vacuo. The crude TFA
salt residue was dissolved in 7 mL of dry methylene chloride and
the pH of the solution was adjusted to pH 8 with 1N NaOH. 56 mg of
a mixture of 4-fluoro-3-acetamidobenzene sulfonylchloride and
3-fluoro-4-acetamidobenz- ene sulfonylchloride (.about.1:1) was
added and the mixture stirred vigorously for 3 hours after which an
additional 25 mg was added and the reaction allowed to continue for
an additional 12 hours. The reaction was then diluted with 50 mL of
ethylene chloride and the organic layer was washed sequentially
with water and brine, dried over MgSO.sub.4 and concentrated in
vacuo. The crude residue was purified using a silica gel flash
chromatography column using a gradient of 3% to 5% MeOH in
methylene chloride as eluant to yield 60 mg of the title compounds.
TLC: Rf=0.50, 10% MeOH/CH.sub.2Cl.sub.2; HPLC: Rt=13.93 min. NMR
(CDCl.sub.3): .delta. 9.05 (s, 1H); 8.65 (d, 0.5H); 8.58 (t, 0.5H),
8.20 (dd, 0.5H), 7.85 (d, 1H) 7.75 (m, 0.5H), 7.45-7.63 (m, 1.5H),
7.14-7.25 (m, 6H), 6.78-6.95 (m, 5H), 6.70 (d, 1H), 6.41 (s, 0.5H),
6.25 (s, 0.5H), 6.18 (s, 0.SH), 6.10 (s, 0.5H), 4.88 (m, 0.5H),
4.81 (m, 0.5H), 4.37 (d, 1H), 4.35 (m, 1H), 4.21 (d, 1H), 4.00 (m,
1H), 3.46 (m, 0.5H), 3.35 (m, 0.5H), 3.27 (d, 0.5H), 3.16 (d,
0.5H), 3.14 (d, 1H), 2.45-2,75 (m, 5H); 2.16, 2.20 (2 s, 3H
total).
EXAMPLE 3
[0322] Compound 3.
[0323] A 23 mg portion of the resultant compound of Example 1E was
treated with 1 mL of 90% aqueous TFA and allowed to stand for 15 h.
The mixture was concentrated in vacuo and the residue taken upon in
2 mL of CH.sub.2Cl.sub.2, treated with 6 .mu.L of DIEA, and cooled
to 0.degree. C. To this solution was added 23 mg of
3,5-dimethylisoxazole-4-sulfonyl chloride, and the mixture was
stirred for 18 h, warming slowly to ambient temperature. After
concentration of the mixture in vacuo, the residue was purified by
preparative reversed-phase C.sub.18HPLC using a linear gradient of
35% to 100% CH.sub.3CN/H.sub.2O with 0.1% TFA for elution to obtain
1.1 mg of the title compound. TLC: Rf=0.55, 10%
MeOH/CH.sub.2Cl.sub.2, HPLC: Rt=14.5 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 4
[0324] Compound 4.
[0325] A 33 mg portion of the resultant compound of Example 1E was
treated with 1 mL of 90% aqueous TFA and allowed to stand for 15 h.
The mixture was concentrated in vacuo and the residue taken up in 3
mL of CH.sub.2Cl.sub.2, treated with 16 .mu.L of DIEA, and cooled
to 0.degree. C. To this solution was added 10 .mu.L of
3-trifluoromethylbenzene sulfonyl chloride, and the mixture was
stirred for 18 h, warming slowly to ambient temperature. After
concentration of the mixture in vacuo, the residue was purified by
preparative reversed-phase C.sub.18HPLC using a linear gradient of
35% to 100% CH.sub.3CN/H.sub.2O with 0.1% TFA for elution to obtain
1.1 mg of the title compound. TLC: Rf=0.55, 10%
MeOH/CH.sub.2Cl.sub.2, HPLC: Rt=14.5 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 5
[0326] Compound 5.
[0327] A 20 mg portion of the resultant compound of Example 1E was
treated with 1 mL of 90% aqueous TFA and allowed to stand for 18 h.
The mixture was concentrated in vacuo and the residue taken up in 1
mL of CH.sub.2Cl.sub.2, treated with 10 .mu.L of DIEA, and cooled
to 0.degree. C. To this solution was added 13 mg of
2-acetamido-4-methyl-5-thiazolesul- fonyl chloride, and the mixture
was stirred for 17 h, warming slowly to ambient temperature. After
concentration of the mixture in vacuo, the residue was purified by
preparative reversed-phase C.sub.18HPLC using a linear gradient of
35% to 100% CH.sub.3CN/H.sub.2O with 0.1% TFA for elution to obtain
0.40 mg of the title compound. TLC: Rf=0.5, 10%
MeOH/CH.sub.2Cl.sub.2, HPLC: Rt=13.8 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 6
[0328] Compound 6.
[0329] A 33 mg portion of the resultant compound of Example 1E was
treated with 1 mL of 90% aqueous TFA and allowed to stand for 16 h.
The mixture was concentrated in vacuo and the residue taken up in 2
mL of CH.sub.2Cl.sub.2, treated with 16 .mu.L of DIEA, and cooled
to 0.degree. C. To this solution was added 11 mg of
5-(isoxazol-3-yl)thiophene-2-sulfo- nyl chloride, and the mixture
was stirred for 18 h, warming slowly to ambient temperature. After
concentration of the mixture in vacuo, the residue was purified by
preparative reversed-phase C.sub.18HPLC using a linear gradient of
35% to 100% CH.sub.3CN/H.sub.2O with 0.1% TFA for elution to obtain
1.5 mg of the title compound. TLC: Rf=0.7, 10%
MeOH/CH.sub.2Cl.sub.2, HPLC: Rt=14.7 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 7
[0330] Compound 7.
[0331] A 35.5 mg portion of the resultant compound of Example 1E
was treated with 1 mL of 90% aqueous TFA and allowed to stand for
18 h. The mixture was concentrated in vacuo and the residue taken
up in 3 mL of CH.sub.2Cl.sub.2, treated with 16 .mu.L of DIEA, and
cooled to 0.degree. C. To this solution was added 10 mg of
3-chlorosulfonylbenzoic acid, and the mixture was stirred for 16 h,
warming slowly to ambient temperature. After concentration of the
mixture in vacuo, the residue was purified by preparative
reversed-phase C.sub.18HPLC using a linear gradient of 35% to 100%
CH.sub.3CN/H.sub.2O with 0.1% tTFA for elution to obtain 1.6 mg of
the title compound. TLC: Rf=0.7, 10% MeOH/CH.sub.2Cl.sub.2, HPLC:
Rt=13.6 min; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 8
[0332] Compound 8.
[0333] 0.04 mmol of the resultant compound of Example 10A was
converted to the free base by partitioning between EtOAc and sat.
NaHCO.sub.3. Treatment of the resulting compound with an excess of
1% HCl/MeOH and concentration in vacuo yielded the hydrochloride
salt as a white solid. This compound was suspended in
CH.sub.2Cl.sub.2 and treated with sufficient DIEA to bring the pH
to >10 (moist pH paper). The solution was treated with 7 molar
equivalents of chlorotrimethylsilane and stirred for 15 h under
nitrogen, then treated with 0.06 mmol of methane sulfonyl chloride
and stirred for 1 h. The resulting mixture was concentrated to a
small volume, applied directly to a thick layer silica gel plate
and eluted with 7% MeOH/CH.sub.2Cl.sub.2. The primary UV-quenching
band was isolated and further purified by preparative
reversed-phase HPLC to yield the title compound as a white solid.
TLC: Rf=0.65, 10% CH.sub.3OH/CH.sub.2Cl.sub.2, HPLC: Rt=12.3 min;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLES 9 AND 192
[0334] A. Compound XIV ((syn, Anti-OH, A=Quinoline-2-Carbonyl,
D'=Isobutyl).
[0335] A solution of 317 mg (0.425 mmol) of the resultant compounds
of Example 17B, diastereomer B and 0.11 mL (0.637 mmol) of
diisopropylethyl amine in 7 mL of dichloromethane was treated with
139.1 mg (0.637 mmol) of di-tert-butyl dicarbonate. After 24 hours,
the mixture was diluted with dichloromethane. The mixture was
washed with water, 5% NaHCO.sub.3, 0.5 N HCl, brine then dried over
MgSO.sub.4, filtered and concentrated in vacuo. The residue was
purified by low pressure silica gel column chromatography using a
20% ethyl acetate/dichloromethane as eluent to yield 81.2 mg of the
fast moving hydroxyl diastereomer, 65.8 mg of the slower moving
hydroxyl diastereomer, and 65.8 mg of the mixed diastereomers. TLC:
Rf=0.60, 0.67, 40% EtOAc/CH.sub.2Cl.sub.2; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
[0336] B. Compounds 9 and 192.
[0337] A solution of 35.1 mg (0.041 mmol) of the resultant mixed
diastereomers (.about.1:1) of Example 9/192A in 0.8 mL of
dichloromethane was treated with 0.8 mL of trifluoroacetic acid.
After 4 hours, the mixture was concentrated in vacuo. TLC: Rf=0.11,
10% CH.sub.3OH/CH.sub.2Cl.sub.2. To a solution of the resulting
trifluoroacetic acid salt (entire yield) in 1 mL of dichloromethane
was sequentialled added 0.3 mL of saturated NaHCO.sub.3, a small
amount of solid NaHCO.sub.3 and 11.8 mg (0.054 mmol) of
benzofurazan-4-sulphonyl chloride. After 3 hours, the mixture was
diluted with dichloromethane. The two layers were separated and the
aqueous layer was extracted once with dichloromethane. The combined
organic layer was washed with brine then dried over MgSO.sub.4,
filtered and concentrated in vacuo. The residue was purified by
preparative HPLC to yield 2.0 mg of compound 9 as a white solid:
TLC: Rf=0.20, 5% CH.sub.3OH/CH.sub.2Cl.sub.2; HPLC, RT 14.2 min.
2.7 mg of compound 192 was also obtained as a white solid, which
was determined by NMR and HPLC to be contaminated with .about.25%
of compound 9: TLC: Rf=0.20, 5% CH.sub.3OH/CH.sub.2Cl.sub.2; HPLC,
Rt=14.2 min. (.sup.1H)-NMR consistent with structure.
EXAMPLE 10
[0338] A. Compound XV ((syn)-OH, A=Quinoline-2-Carbonyl, D'=Benzyl;
TFA Salt).
[0339] A 0.degree. C. solution of 1.027 g portion of the resultant
compound of Example 1E in 5 mL of CH.sub.2Cl.sub.2 was treated with
5 mL of TFA and allowed to stand for 3 h. The mixture was
concentrated in vacuo to yield 0.95 g of the title compound, which
was used without subsequent purification.
[0340] B. Compound 10.
[0341] A solution of 30.2 mg of the resultant compound of Example
10A in 3 mL of CH.sub.2Cl.sub.2 was treated with 0.33 mL of DIEA
and 31.1 mg of m-benzenedisulfonyl chloride. The mixture was
stirred for 2 h, then treated with 2 mL of concentrated aqueous
ammonium hydroxide. The biphasic mixture was stirred for an
additional 16 h, concentrated in vacuo, and the residue partitioned
between ethyl acetate and brine. The organic layer was dried over
anhydrous MgSO.sub.4 and concentrated in vacuo, and the residue was
purified by preparative thick layer silica gel chromatography using
3% MeOH/CH.sub.2Cl.sub.2 as eluant to yield 4.5 mg of the title
compound. TLC: Rf=0.5, 3% MeOH/CH.sub.2Cl.sub.2 as eluant to yield
4.5 mg of the title compound. TLC: Rf=0.5, 3%
MeOH/CH.sub.2Cl.sub.2, HPLC: Rt=13.4 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 11
[0342] Compound 11.
[0343] A solution of 57.9 mg of the resultant compound of Example
10A in 5 mL of CH.sub.2Cl.sub.2 was treated with 30 .mu.L of DIEA
and 9.3 .mu.L of dimethylsulfamoyl chloride. The mixture was
stirred for 12 h, then treated with an additional 30 .mu.L of DIEA
and 9.3 .mu.L of dimethylsulfamoyl chloride and the reaction was
allowed to proceed an additional 12 hours. The mixture was then
diluted with CH.sub.2Cl.sub.2 and washed with saturated NH.sub.4Cl;
the aqueous layer was washed with CH.sub.2Cl.sub.2, and the
combined organic extracts were dried over MgSO.sub.4. Filtration
and concentration provided a residue which was chromatographed on a
silica gel column using 2.5% MeOH/EtOAc as eluent, yielding a
slightly impure product which was further purified by preparative
HPLC using a linear gradient of 35% to 100% CH.sub.3/CN/H.sub.2O
with 0.1% TFA for elution. HPLC: Rt=13.0 minutes. NMR (CDCL.sub.3):
.delta. 9.15 (d, 1H), 8.34 (d, 1H), 8.22 (d, 1H), 8.18 (d, 1H),
7.90 (d, 1H), 7.80 (t, 1H) 7.65 (t, 1H), 7.16-738 (m, 5H), 7.05 (d,
1H), 6.95 (t, 1H), 6.87 (t, 1H), 5.85 (br s, 1H), 5.62 (br s, 1H),
4.87 (M, 1H), 4.46 (s, 2H), 4.08 (m, 1H), 3.66 (m, 1H), 3.30 (m,
2H), 2.59-2.94 (m, 4H), 2.81 (s, 6H).
EXAMPLE 12
[0344] A. Compound XIV ((syn)-OH, A=Quinoline-2-Carbonyl,
D'=Benzyl; Trifluoroacetate Salt).
[0345] To a solution of 1.027 g (1.164 mmol) of the resultant
compound of Example 1E in CH.sub.2Cl.sub.2 (5 mL) at 0.degree. to
5.degree. C. was added trifluoromethanesulfonic acid (5 mL). After
stirring for 3 h, the reaction mixture was concentrated in vacuo to
provide 0.95 g of light yellow, gummy product, containing one
equivalent of triphenylmethanol, which was used without subsequent
purification.
[0346] B. Compound 12.
[0347] To a solution of 30.2 mg (0.038 mmol) of the resultant
compound of Example 12A in CH.sub.2Cl.sub.2 (3 mL) was added
diisopropylethylamine (0.33 mL, 0.189 mmol), and
2-(pyrid-2-yl)-tyiophene-5-sulfonyl chloride 13 mg, (0.249 mmol).
After 14 h, the resulting mixture was diluted with ethyl acetate,
washed with saturated brine, dried over magnesium sulfate, filtered
and concentrated in vacuo. The residue was purified by preparative
reversed-phase chromatography using a 5% to 100%
H.sub.2O/acetonitrile gradient as eluant to yield the title
product.
EXAMPLE 13
[0348] Compound 13.
[0349] To a solution of 30 mg (0.038 mmol) of the resultant
compound of Example 12A in CH.sub.2Cl.sub.2 (3 mL) was added
diisopropylethylamine (0.33 mL, 0.189 mmol), and
2-(3-phenylsulfonyl)thiophene sulfonyl chloride (0.113 mmol). After
stirring for 2 h, the reaction mixture was made biphasic by
addition of 30% ammonium hydroxide solution (2 mL). After stirring
for an additional 16 h, the resultant mixture was concentrated in
vacuo, reconstituted in ethyl acetate, washed with saturated brine,
dried over magnesium sulfate, filtered, and re-concentrated in
vacuo. Purification by thin layer preparative chromatography
yielded the desired compound.
EXAMPLE 14
[0350] Compound 14.
[0351] The resulting compound of Example 17B, diastereomer B (170
mg) was treated with 1 mL of 90% aqueous TFA and allowed to stand
for 12 h. The mixture was concentrated in vacuo and the residue
taken up in 5 mL of dry CH.sub.2Cl.sub.2. To this solution, 3 mL of
saturated aqueous sodium bicarbonate and 50 mg of
4-fluorobenzenesulfonyl chloride was added and the mixture stirred
for 3 h. The resulting mixture was diluted with CH.sub.2Cl.sub.2
and washed with water, dried over magnesium sulfate and filtered.
After concentration of the mixture in vacuo, a portion of the
residue was purified by preparative reversed-phase C.sub.18HPLC
using a linear gradient of 35% to 100% CH.sub.3CN/H.sub.2O with
0.1% TFA for elution to obtain 3.0 mg of the title compound. TLC:
Rf=0.25, 5% CH.sub.3OH in CH.sub.2Cl.sub.2, HPLC: Rt=14.78 min;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 15
[0352] Compound 15.
[0353] A sample of a mixture of 4-fluoro-3-acetamidobenzenesulfonyl
chloride and 3-fluoro-4-acetamidobenzenesulfonyl chloride (approx.
1:1; obtained from Maybridge Chemicals) was resolved into its
respective regioisomers by silica gel chromatography using 10%
isopropyl alcohol/hexane as eluent. A solution of
4-acetamido-3-fluorobenzenesulfon- yl chloride (30 mg) and the
resulting compound of Example 17B, diastereomer B (80 mg) in 10 mL
of CH.sub.2Cl.sub.2 was reacted in the same manner as described for
Example 14. After workup and purification of a portion of the
product by preparative reversed-phase C.sub.18HPLC using a linear
gradient of 35% to 100% CH.sub.3CN/H.sub.2O with 0.1% TFA as
eluent, 1.2 mg of the title compound was obtained as a white solid.
TLC: Rf=0.25, 5% CH.sub.3OH in CH.sub.2Cl.sub.2, HPLC: Rt=12.91
min; (.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 16
[0354] Compound 16.
[0355] 80 mg of the resulting compound of Example 17B, diastereomer
B, was reacted with 45 mg of 3-acetamido-4-fluorobenzenesulfonyl
chloride in the same manner as described for Example 14. After
workup and purification of a portion of the product by preparative
reversed-phase C.sub.18HPLC using a linear gradient of 35% to 100%
CH.sub.3CN/H.sub.2O with 0.1% TFA as eluent, 1.4 mg of the title
compound was obtained. TLC: Rf=0.25, 5% CH.sub.3OH in
CH.sub.2Cl.sub.2, HPLC: Rt=12.91 min; (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 17
[0356] A. (2S)-2-((1S, 2R syn,
anti)-3-(2-methylpropyl)amino-1-benzyl-2-hy-
doxypropyl)-N.sup.1-((quinoline-2-carbonyl)-amino)-N.sup.4-trityl
succinamide.
[0357] A solution of 683.1 mg (0.96 mmol) of the resultant
compounds of Example 191D and 1.9 mL (19.2 mmol) of isobutylamine
in 10 mL of acetonitrile in a sealed tube was heated at
90-100.degree. C. for 24 hours. After cooling to room temperature,
the mixture was concentrated in vacuo. The residue was taken up in
dichloromethane and washed with water, brine, then dried over
MgSO.sub.4, filtered and concentrated in vacuo to yield 783.8 mg of
the mixed diastereomeric products. TLC: Rf=0.11, 10%
CH.sub.3OH/CH.sub.2Cl.sub.2; (.sup.1H)-NMR (CDCl.sub.3) consistent
with structure.
[0358] B. Compound XIII, ((syn, anti)-OH, A=Quinoline-2-Carbonyl,
D'=Isobutyl).
[0359] A solution of 583.8 mg of the resultant compounds of Example
17A and 0.2 mL of diisopropylethylamine in 10 mL of dichloromethane
was treated with 256 mg of di-tert-butyl dicarbonate. After 24
hours, the mixture was diluted with dichloromethane. The mixture
was washed with water, 5% NaHCO3, 0.5 N HCl, brine then dried over
MgSO.sub.4, filtered and concentrated in vacuo. The residue was
purified by low pressure silica gel column chromatography using a
20% ethyl acetate/dichloromethane as eluent to yield 154.6 mg of
the fast moving diastereomer A, later identified as having the anti
configuration at the hydroxyl center; 98.8 mg of the slower moving
diastereomer B, having the syn configuration at the hydroxyl
center, and 204.6 mg of the mixed diastereomers A and B. TLC:
Rf=0.60, 0.67, 40% EtOAc/CH.sub.2Cl.sub.2.
[0360] C. Compound 17.
[0361] A solution of 64.6 mg of the resultant compounds of Example
17B, diastereomer B, in 1.5 mL of dichloromethane was treated with
1.5 mL of trifluoroacetic acid. After 4 hours, the mixture was
concentrated in vacuo to yield the amine trifluoroacetate salt.
TLC: Rf=0.11, 10% CH.sub.3OH/CH.sub.2Cl.sub.2. To a solution of
17.8 mg of the resultant trifluoroacetate salt in 1 mL of
dichloromethane was sequentially added 0.3 mL of saturated
NaHCO.sub.3, a small amount of solid NaHCO.sub.3 and 10.7 mg of
4-acetamido-benzenesulphonyl chloride. After 3 hours, the mixture
was diluted with dichloromethane. The two layers were separated and
the aqueous layer was extracted once with dichloromethane. The
combined organic layer was washed with brine then dried over
MgSO.sub.4, filtered and concentrated in vacuo. The residue was
purified by preparative HPLC to yield 14.4 mg of the title compound
as a white solid; TLC: Rf=0.54, 10% CH.sub.3OH/CH.sub.2Cl.sub.2;
HPLC, Rt=13.58 min; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 18
[0362] Compound 18.
[0363] To a solution of 20.8 mg (0.041 mmol) of the crude
trifluoroacetate salt obtained as from Example 17B, diastereomer B,
in 1 mL of dichloromethane was sequentially added 0.3 mL of
saturated NaHCO.sub.3, a small amount of solid NaHCO.sub.3 and 13.6
mg (0.054 mmol) of 2-acetamido-4-methyl-5-thiazolesulphonyl
chloride. After 3 hours, the mixture was diluted with
dichloromethane. The two layers were separated and the aqueous
layer was extracted once with dichloromethane. The combined organic
layer was washed with brine then dried over MgSO.sub.4, filtered
and concentrated in vacuo. The residue was purified by preparative
HPLC to yield 4.8 mg of the title compound as a white solid; TLC:
Rf=0.50, 10% CH.sub.3OH/CH.sub.2Cl.sub.2; HPLC: Rt=13.35 min;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 19
[0364] A. Sodium 3-Acetamidobenzenesulfonate.
[0365] A solution of 118.6 mg (0.55 mmol) of
3-acetamidobenzenesulfonic acid in 0.5 mL of water was treated with
0.55 mL (0.55 mmol) of 1.0 N NaOH at 0.degree. C. After stirring at
room temperature for 4 hours, the mixture was concentrated to
dryness and used without subsequent purification.
[0366] B. 3-Acetamidobenzenesulfonyl Chloride.
[0367] The crude mixture from Example 19A was cooled to 0.degree.
C. and 0.29 g (1.38 mmol) of phosphorus pentachloride was added.
The mixture of solid was stirred for 3 hours then 5 mL
dichloromethane was added. After 24 hours, the slurry was filtered
and concentrated in vacuo to yield 81.4 mg of solid product which
was used without subsequent purification. TLC: Rf=0.50, 40%
EtOAC/CH.sub.2Cl.sub.2.
[0368] C. Compound 19.
[0369] A solution of 82.7 mg (0.098 mmol) of diastereomer B,
obtained in Example 17B, in 2 mL of dichloromethane was treated
with 2 mL of trifluoroacetic acid. After 4 hours, the mixture was
concentrated in vacuo to yield the amine trifluoroacetate salt
which was used without further purification; TLC: Rf=0.11, 10%
CH.sub.3OH/CH.sub.2Cl.sub.2. A solution of this salt (entire yield)
in 2 mL of dichloromethane was treated sequentially with 0.5 mL of
saturated NaHCHO.sub.3, small amount of solid NaHCO.sub.3 and a
solution of 81.4 mg (0.046 mmol) of the resultant compound of
Example 19B. After 3 hours, the mixture was diluted with
dichloromethane. The two layers were separated and the aqueous
layer was extracted once with dichloromethane. The combined organic
layer was washed with brine then dried over MgSO.sub.4, filtered
and concentrated in vacuo. The residue was purified by preparative
HPLC to yield 24.7 mg of the title compound as a white solid; TLC:
Rf=0.42, 10% CH.sub.3OH/CH.sub.2Cl.sub.2; HPLC: Rt=13.8 min;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 20
[0370] Compound 20.
[0371] A solution of 209.0 mg (0.24 mmol) of the resultant compound
of Example 17B, diastereomer B, in 5 mL of dichloromethane was
treated with 5 mL of trifluoroacetic acid. After 4 hours, the
mixture was concentrated in vacuo. TLC: Rf=0.11, 10%
CH.sub.3OH/CH.sub.2Cl.sub.2. To a solution of this residue in 2 mL
of dichloromethane was sequentially added 0.5 mL of saturated
NaHCO3, a small amount of solid NaHCO.sub.3 and 70.2 mg (0.32 mmol)
of benzofurazan-4-sulphonyl chloride. After 3 hours, the mixture
was diluted with dichloromethane. The two layers were separated and
the aqueous layer was extracted once with dichloromethane. The
combined organic layer was washed with brine then dried over
MgSO.sub.4, filtered and concentrated in vacuo. The residue was
purified by preparative HPLC to yield 108.0 mg of the title
compound as a white solid; TLC: Rf=0.60, lot
CH.sub.3OH/CH.sub.2Cl.sub.2; HPLC: Rt=14.95 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 21
[0372] Compound 21.
[0373] The resulting compound of Example 17B, diastereomer B, (228
mg, 0.27 mmol) was dissolved in 1:1 CH.sub.2Cl.sub.2/TFA (10 mL),
and the reaction mixture stirred for 3.5 hours, then concentrated
to dryness to afford the product trifluoroacetate salt as a yellow
solid which was used in the next reaction without purification. To
a solution of this residue (34.7 mg, 0.05 mmol) in CH.sub.2Cl.sub.2
(3 mL) was added Heunig's base (41 .mu.l, 0.24 mmol) and
dimethylsulfamoyl chloride (11 .mu.l, 0.09 mmol), and the reaction
was stirred for 17 hours at room temperature. The reaction mixture
was then diluted with CH.sub.2Cl.sub.2 and washed with saturated
NH.sub.4Cl, and the organic layer was dried over MgSO.sub.4.
Filtration and concentration provided a residue which was
chromatographed on a silica gel column using 8%
CH.sub.3OH/CH.sub.2Cl.sub.2 as eluent, yielding the desired
compound which was further subject to purification by preparative
HPLC. HPLC: Rt=13.8 minutes. TLC: Rf=0.40, 8%
CH.sub.3OH/CH.sub.2Cl.sub.2; (.sup.1H)-NMR (CDCl.sub.3) consistent
with structure.
EXAMPLE 22
[0374] A. N.sup..alpha.-isocyano-L-valine Methyl Ester.
[0375] To the HCl salt of valine methyl ester (2.08 g, 12.40 mmol)
in toluene (20 mL) was added a 20% solution of phosgene in toluene
(32 mL, 62.00 mmol), and the solution was heated at reflux for 12
hours. The reaction was then cooled to room temperature and
concentrated in vacuo to give a pale yellow liquid which was used
in the subsequent reaction without purification. TLC: Rf=0.88, 50%
Hexane/EtOAc; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
[0376] B. N.sup..alpha.-(2-pyridylmethyl)-oxycarbonyl-L-valine
Methyl Ester.
[0377] A mixture of 2-pyridylcarbinol (941 .mu.l, 9.75 mmol) and
the resulting compound of Example 22A (1.28 g, 8.12 mmol) were
allowed to stir in CH.sub.2Cl.sub.2 (7 mL) for 12 hours, then the
reaction was concentrated and the residue chromatographed with 50%
hexane/EtOAc to afford 2.03 grams of the title compound as a
colorless oil. TLC: Rf=0.26, 50% Hexane/EtOAc; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
[0378] C. N.sup..alpha.-(2-pyridylmethyl)-oxycarbonyl-L-valine.
[0379] A solution of the resulting compound of Example 22B (634 mg,
2.38 mmol) in a 1/1 mixture of 1N HCl/THF (6 mL) containing 12 N
HCl (0.5 mL) was allowed to stir at room temperature over 15 hours,
but much starting material was still present by TLC. Hence, more 12
N HCL was added (1 mL), and the reaction stirred an additional 48
hours. The reaction was then concentrated to dryness and diluted
with CH.sub.2Cl.sub.2, yielding the desired carboxylic acid as an
insoluble resin which was washed with additional CH.sub.2Cl.sub.2,
providing 22C which contained minor quantities of 22B. This
material was used in the subsequent reaction without further
purification. TLC: Rf=0.11, 8% CH.sub.3OH/CH.sub.2Cl.sub.- 2;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
[0380] D. Compound XXX (A=(2-pyridylmethyl)-oxycarbonyl,
R.sup.3=Isopropyl, R.sup.3'=H, D'=Isobutyl,
A'=Tert-butoxycarbonyl).
[0381] To the resulting compound of Example 21B (277 mg, 0.82 mmol)
in CH.sub.2Cl.sub.2 (5 mL) was added
1-(3-dimethylaminopropyl)-3-ethylcarbod- iimide hydrochloride (210
mg, 1.10 mmol), the acid 22C (402 mg, 1.10 mmol), and
1-hydroxybenzotriazole hydrate (148 mg, 1.10 mmol). The reaction
proceeded for 12 hours at room temperature, then was diluted with
CH.sub.2Cl.sub.2 and washed successively with saturated NH.sub.4Cl
and NaHCO.sub.3, and the organic layer was dried over MgSO.sub.4.
Filtration and concentration provided a residue which was
chromatographed on a silica gel column using 17%
THF/CH.sub.2Cl.sub.2 as eluent, yielding 396 mg of product. TLC:
Rf=0.26, 17% THF/CH.sub.2Cl.sub.2; (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
[0382] E. Compound 22. The resulting compound of Example 22D (396
mg, 0.69 mmol) was dissolved in 90% aqueous TFA (11 mL), and the
reaction mixture stirred for 3 hours at room temperature, then was
concentrated to dryness. To a solution of this residue (231 mg.
0.33 mmol) in CH.sub.2Cl.sub.2 (5 mL) was added excess solid
NaHCO.sub.3 (approx. 1 gram) and saturated aqueous NaHCO.sub.3 (20
.mu.l), followed by N-acetylsulfanilyl chloride (116 mg, 0.50
mmol), and the reaction proceeded for 12 hours at room temperature.
The reaction mixture was then diluted with CH.sub.2Cl.sub.2 and
washed with saturated NaHCO.sub.3, and the organic layer was dried
over MgSO.sub.4. Filtration and concentration provided a residue
which was chromatographed on a silica gel column using 8%
CH.sub.3OH/CH.sub.2Cl.sub.2 as eluent, yielding the desired
compound which was further subject to purification by preparative
HPLC (76.1 mg of 3 was obtained). HPLC: Rt=12.1 minutes. TLC:
Rf=0.46, 8% CH.sub.3OH/CH.sub.2Cl.sub.2; NMR (CDCl.sub.3): 8.76 (d,
1H), 8.40 (br s, 1H), 8.26 (t, 1H), 7.72 (d, 2H), 7.67 (d, 2H),
7.58 (d, 2H), 7.37 (d, 1H), 7.25 (m, 4H), 7.16 (br d, 1H), 6.47 (d,
1H), 5.65 (d, 1H), 5.26 (d, 1H), 4.32 (m, 1H), 3.91 (t, 1H), 3.83
(m, 1H), 3.23 (d, 1H), 3.05 (m, 2H), 2.68-3.10 (m, 3H), 2.22 (m,
3H), 2.0 (m, 1H), 1.82 (m, 1H), 0.85 (d, 3H), 0.80 (d, 3H), 0.71
(d, 3H), 0.65 (d, 3H).
EXAMPLE 23
[0383] Compound 23.
[0384] Prepared by the same route as described for Example 22,
except 4-pyridylcarbinol was utilized for reaction with the product
of Example 22A. HPLC: Rt=12.0 minutes. TLC: Rf=0.50 (8%
CH.sub.3OH/CH.sub.2Cl.sub.2)- ; (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 24
[0385] Compound 24.
[0386] A solution of the resulting compound of the trifluoroacetic
acid deprotection of Example 22D (as described in Example 22E; 215
mg, 0.31 mmol) in CH.sub.2Cl.sub.2 at room temperature was treated
with diisopropylethylamine (214 .mu.l, 1.23 mmol) and
dimethylsulfamoyl chloride (40 .mu.l, 0.37 mmol) in
CH.sub.2Cl.sub.2 at room temperature in CH.sub.2Cl.sub.2 at room
temperature for 12 hours. The reaction mixture was concentrated and
chromatographed on a silica gel column with 5%
CH.sub.3OH/CH.sub.2Cl.sub.2 as eluent, yielding the desired
compound which was further subject to purification by preparative
HPLC (9.5 mg obtained). HPLC: Rt=14.4 minutes. TLC: Rf=0.88, 11%
CH.sub.3OH/CH.sub.2Cl.sub.2; (.sup.1H)-NMR (CDCl.sub.3) consistent
with structure.
EXAMPLE 25
[0387] Compound 25.
[0388] This compound was prepared by the route described for
Example 22, except that 3-pyridylcarbinol was utilized for reaction
with the compound produced in Example 22A, and in the reaction
corresponding to 22E, the trifluoracetate-deprotected material was
reacted with benzofurazan-4-sulphonyl chloride. HPLC: Rt=9.4
minutes. TLC: Rf=0.10, 11% CH.sub.3OH/CH.sub.2Cl.sub.2;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 26
[0389] Compound 26.
[0390] A solution of the resulting compound from the
trifluoroacetic acid deprotection of Example 22D (as described in
Example 22E; 27 mg, 0.14 mmol) in CH.sub.2Cl.sub.2 was treated with
excess solid NaHCO.sub.3 (approx. 1 gram) and saturated aqueous
NaHCO.sub.3 (7 .mu.l), then stirred vigorously at room temperature
for 3 hours. The reaction mixture was decanted from the solids,
concentrated, then the residue was purified directly by preparation
HPLC (3.0 mg of white solid obtained). HPLC: Rt=14.7 minutes;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 27
[0391] Compound 27.
[0392] A solution of 33 mg of the resultant compound of Example 40A
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 20 mg of
N,N-diisopropylethylamine and 9.3 mg of allyl chloroformate. The
mixture was stirred for 3 h and then concentrated in vacuo. The
residue was taken up in ethyl acetate and washed with 0.5 N HCl and
saturated NaCl then dried over MgSO.sub.4, filtered, and
concentrated in vacuo. The residue was purified by preparative thin
layer silica gel chromatography using a 2:1 mixture of (5:10:85
NH.sub.4OH/CH.sub.3OH/CH.sub.2Cl.sub.2):diethyl ether to yield 24
mg of the title compound as a white solid. TLC: Rf=0.53, 5:10:85
NH.sub.4OH/CH.sub.3OH/CH.sub.2Cl.sub.2. HPLC: Rt=14.53 min;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 28
[0393] Compound 28.
[0394] A solution of 47.5 mg of the resultant compound of Example
40A in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 28.7 mg of
N,N-diisopropylethylamine and 15.2 mg of isobutyl chloroformate.
The mixture was stirred 3 h and then concentrated in vacuo. The
residue was taken up in ethyl acetate and washed with 0.5 N HCl and
saturated NaCl then dried over MgSO.sub.4, filtered, and
concentrated in vacuo. The residue was purified by preparative thin
layer silica gel chromatography using a 2:1 mixture of (5:10:85
NH.sub.4OH/CH.sub.3OH/CH.sub.2Cl.sub.2):diethyl ether to yield 45
mg of the title compound as a white solid. TLC: Rf=0.60, 5:10:85
NH.sub.4OH/CH.sub.3OH/CH.sub.2Cl.sub.2, HPLC: Rt=15.58 min;
(1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 29
[0395] Compound 29.
[0396] A solution of 35.6 mg of the resultant compound of Example
40A in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 21.5 mg of
N,N-diisopropylethylamine and 0.083 nL of 1.0 M isopropyl
chloroformate. The mixture was stirred 3 h and then concentrated in
vacuo. The residue was taken up in ethyl acetate and washed with
0.5 N HCl and saturated NaCl then dried over MgSO.sub.4, filtered,
and concentrated in vacuo. The residue was purified by preparative
thin layer silica gel chromatography using a 2:1 mixture of 5:10:85
NH.sub.4OH/CH.sub.3OH/CH.sub.2Cl.sub.2:diethyl ether to yield 33.2
mg of the title compound as a white solid. TLC: Rf=0.56, 5:10:85
NH.sub.4OH/CH.sub.3OH/CH.sub.2Cl.sub.2, HPLC: Rt=14.81 min;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 30
[0397] A. (2-Pyrrolidinonyl-hydroxyethyl-N-hydroxysuccinimdyl
Carbonate.
[0398] A solution of 572 mg of 1-(2-hydroxyethyl)-2-pyrrolidinone
and 1.70 g of N,N'-disuccinimidyl carbonate in acetonitrile was
treated, at ambient temperature under an atmosphere of nitrogen,
with 1717 mg of N,N-diisopropylethylamine. The mixture was stirred
for 14 h and concentrated in vacuo. The residue was taken up in
ethyl acetate and washed with saturated NaHCO.sub.3, saturated
NaCl, then dried over MgSO.sub.4, filtered, and concentrated in
vacuo to yield 200 mg of a white solid. TLC: Rf=0.56, 10%
isopropanol in CH.sub.2Cl.sub.2; (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
[0399] B. Compound 30.
[0400] A solution of 68 mg of the resultant compound of Example 30A
in CH.sub.2Cl.sub.2 was added, at ambient temperature under an
atmosphere of nitrogen, to a solution of 32 mg of the resultant
compound of Example 40A and 39 mg N,N-diisopropylethylamine in
CH.sub.2Cl.sub.2. The mixture was stirred for 4 h, diluted with
CH.sub.2Cl.sub.2, washed with saturated NaRCO.sub.3 and saturated
NaCl, then dried over MgSO.sub.4, filtered, and concentrated in
vacuo. The residue was subjected to preparative thin layer silica
gel chromatography using a 2:1 mixture of 5:10:85
NH.sub.4OH/CH.sub.3OH/CH.sub.2Cl.sub.2:diethyl ether to yield 45 mg
of residue. About 20 mg of this residue was purified by preparative
HPLC to yield 13.5 mg of the title compound as a white solid. TLC:
Rf=0.47, 5:10:85 NH.sub.4OH/CH.sub.3OH/CH.sub.2Cl2.HPLC: Rt=12.79
mid; (1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 31
[0401] Compound 31.
[0402] A solution of 39.7 mg of the resultant compound of Example
40A in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 24 mg of
N,N-diisopropylethylamine and 14.5 mg of phenyl chloroformate. The
mixture was stirred for 3 h and then concentrated in vacuo. The
residue was taken up in ethyl acetate and washed with 0.5 N HCl and
saturated NaCl then dried over MgSO.sub.4, filtered, and
concentrated in vacuo. The residue was purified by preparative thin
layer silica gel chromatography using a 2:1 mixture of 5:10:85
NH.sub.4OH/CH.sub.3OH/CH.sub.2Cl.sub.2:diethyl ether to yield 39.7
mg of the title compound. TLC: Rf=0.53, 5:10:85
NH.sub.4OH/CH.sub.3OH/CH.sub.2Cl.sub.2, HPLC: Rt=15.22 min;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 32
[0403] Compound 32.
[0404] A solution of 391 mg of the resultant compound of Example
39A in 4:1 CH.sub.2Cl.sub.2/saturated aqueous NaHCO.sub.3 was
treated sequentially, at ambient temperature under an atmosphere of
nitrogen, with 271 mg of 4-fluorobenzenesulfonyl chloride and 117
mg of sodium bicarbonate. The mixture was stirred for 14 h, diluted
with CH.sub.2Cl.sub.2, washed with saturated NaCl then dried over
MgSO.sub.4, filtered, and concentrated in vacuo. The residue was
purified by low pressure silica gel chromatography using 5% diethyl
ether in CH.sub.2Cl.sub.2 as eluent to yield 420 mg of the title
compound as a white solid. TLC: Rf=0.20, 5% diethyl ether in
CH.sub.2Cl.sub.2. HPLC: Rt=17.41 min; (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 33
[0405] Compound 33.
[0406] A solution of 30 mg of the resultant compound of Example 40A
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 18.1 mg of
N,N-diisopropylethylamine and 9.3 mg of benzyl isocyanate. The
mixture was stirred 14 h and then concetrated in vacuo. The residue
was taken up in ethyl acetate and washed with 0.5 N HCl and
saturated NaCl then dried over MgSO.sub.4, filtered, and
concentrated in vacuo. The residue was purified by preparative thin
layer silica gel chromatography using a mixture of 5:10:85
NH.sub.4OH/CH.sub.3OH/CH.sub.2Cl.sub.2 to yield 30.2 mg of the
title compound as a white solid. TLC: Rf=0.56, 5:10:85
NH.sub.4OH/CH.sub.3OH/CH.sub.2Cl.sub.2, HPLC: Rt=14.36 min;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 34
[0407] Compound 34.
[0408] A solution of 55 mg of the resultant compound of Example 40A
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 33.3 mg of
N,N-diisopropylethylamine and 17.8 mg of 2-methoxyethyl
chloroformate. The mixture was stirred for 3 h and then
concentrated in vacuo. The residue was taken up in ethyl acetate
and washed with 0.5 N HCl and saturated NaCl then dried over
MgSO.sub.4, filtered, and concentrated in vacuo. The residue was
purified by preparative thin layer silica gel chromatography using
a 2:1 mixture of (5:10:85
NH.sub.4OH/CH.sub.3OH/CH.sub.2Cl.sub.2):diethyl ether to yield 48.1
mg of the title compound as a white solid. TLC: Rf=0.56, 5:10:85
NH.sub.4OH/CH.sub.3OH/CH.sub.2Cl.sub.2. HPLC: Rt=13.43 min;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 35
[0409] A. Compound XXI (D'=Isobutyl, A'=4-Fluorophenyl,
Hydrochloride Salt).
[0410] A solution of 398 mg of the resultant compound of Example 32
in ethyl acetate was treated at -20.degree. C. with HCl gas. The
HCl was bubbled through the mixture for 20 min over which time the
temperature was allowed to warm to 20.degree. C. Nitrogen was then
bubbled through the mixture for 15 min and solvent removed in vacuo
to yield 347 mg of the title compound as a white solid. TLC:
Rf=0.82, 5:10:85 NH.sub.4OH/CH.sub.3OH/CH.sub.2Cl.sub.2;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
[0411] B. Compound 35.
[0412] A solution of 111 mg of the resultant compound of Example
35A in CH.sub.2Cl.sub.2 was added, at ambient temperature under an
atmosphere of nitrogen, to a solution of 118 mg of the resultant
compound of Example 48A and 133 mg N,N-diisopropylethylamine in
CH.sub.2Cl.sub.2. The mixture was stirred for 14 h, diluted with
CH.sub.2Cl.sub.2, washed with saturated NaHCO.sub.3 and saturated
NaCl, then dried over MgSO.sub.4, filtered, and concentrated in
vacuo. The residue was subjected to preparative thin layer silica
gel chromatography using 5% CH.sub.3OH in CH.sub.2Cl.sub.2 to yield
98.8 mg of the title compound as a white solid. TLC: Rf=0.48, 5%
CH.sub.3OH in CH.sub.2Cl.sub.2, HPLC: Rt=15.18 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 36
[0413] Compound 36.
[0414] A solution of 48 mg of the resultant compound of Example 40A
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 29.0 mg of
N,N-diisopropylethylamine and 15.1 mg of 3-butenyl chloroformate.
The mixture was stirred for 3 h and then concentrated in vacuo. The
residue was taken up in ethyl acetate and washed with 0.5 N HCl and
saturated NaCl then dried over MgSO.sub.4, filtered, and
concentrated in vacuo. The residue was purified by preparative thin
layer silica gel chromatography using a 2:1 mixture of (5:10:85
NH.sub.4OH/CH.sub.3OH/CH.sub.2Cl.sub.2):diethyl ether to yield 43.8
mg of the title compound as a white solid. TLC: Rf=0.83, 5:10:85
NH.sub.4OH/CH.sub.3OH/CH.sub.2Cl.sub.2; Rf=0.24, 5% diethyl ether
in CH.sub.2Cl.sub.2, HPLC: Rt=14.76 min; (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 37
[0415] Compound 37.
[0416] A solution of 99 mg of the resultant compound of Example 51D
in 4:1 CH.sub.2Cl.sub.2/saturated aqueous NaHCO.sub.3 was treated
sequentially, at ambient temperature under an atmosphere of
nitrogen, with 83.2 mg of 3,4-dichlorobenzenesulfonyl chloride and
29 mg of sodium bicarbonate. The mixture was stirred for 14 h,
diluted with CH.sub.2Cl.sub.2, washed with saturated NaCl then
dried over MgSO.sub.4, filtered, and concentrated in vacuo. The
residue was subjected to operative thin layer silica gel
chromatography using 5% CH.sub.3OH in CH.sub.2Cl.sub.2 to yield 107
mg of the title compound as a white solid. TLC: Rf=0.35 (5%
CH.sub.3OH in CH.sub.2Cl.sub.2). HPLC: Rt=17.27 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 38
[0417] Compound 38.
[0418] To a solution of 32 mg of the resultant compound of Example
35A in CH.sub.2Cl.sub.2 was added, at ambient temperature under an
atmosphere of nitrogen, 14 mg of benzyl chloroformate and 21 mg
N,N-diisopropylethylami- ne. The mixture was stirred for 4 h,
diluted with CH.sub.2Cl.sub.2, washed with saturated NaHCO.sub.3
and saturated NaCl, then dried over MgSO.sub.4, filtered, and
concentrated in vacuo. The residue was purified by preparative thin
layer silica gel chromatography using 10% diethyl ether in
CH.sub.2Cl.sub.2 as eluent to yield 33 mg of product. TLC: Rf=0.62,
10% diethyl ether in CH.sub.2Cl.sub.2, HPLC: Rt=17.27 min.
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 39
[0419] A. Compound XXI (D'=Isobutyl, A=Tert-butoxy Carbonyl,
A'=H).
[0420] A solution of 4.1 g of epoxide XX (A=Boc) in 30 mL of
ethanol was treated with 22.4 mL of isobutylamine and heated under
reflux for 1 h. The mixture was concentrated to yield the title
compound as a white solid which was used without subsequent
purification. NMR (CDCl.sub.3): .delta. 0.91 (d, 3H); 0.93 (d, 3H);
1.37 (s, 9H); 1.68 (br s, 2H); 2.40 (d, 2H); 2.68 (d, 2H); 2.87
(dd, 1H); 2.99 (dd, 1H); 3.46 (dd, 1H); 3.75 (br s, 1H); 3.80 (br
s, 1H); 4.69 (d, 1H); 7.19-7.32 (m, 4H).
[0421] B. Compound 39.
[0422] To a solution of 514.1 mg of the resultant compound of
Example 39A in dichloromethane (10 mL) was added aqueous sodium
bicarbonate (5 mL) and N-acetylsulfanilyl chloride (428.4 mg).
After 14 h, the resulting mixture was diluted with ethyl acetate,
washed with sodium bicarbonate, saturated brine, dried over
magnesium sulfate, filtered and concentrated in vacuo. The residue
was purified by low pressure silica gel column chromatography using
20% ethyl acetate in dichloromethane eluent to yield 714.4 mg of
the title product. TLC: Rf=0.63, 60% ethyl acetate/dichloromethane,
HPLC: Rt=15.3 min; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 40
[0423] A. Compound XXII (D'=Isobutyl, A=H, E=4-Acetamidophenyl),
Hydrochloride Salt.
[0424] To a solution of 691.4 mg (1.296 mmol) of the resultant
compound of Example 39B in ethyl acetate (20 mL) at -20.degree. C.
was bubbled anhydrous HCl gas for 10 min. The ice bath was removed
and after an additional 15 min., the reaction mixture was sparged
with nitrogen then concentrated in vacuo to provide 610 mg of title
product which was used without subsequent purification.
[0425] B. Compound 40.
[0426] A solution of 41.5 mg of the resultant crude compound of
Example 40A in 5 mL of dichloromethane was treated sequentially, at
ambient temperature under an atmosphere of nitrogen, with 18.1 mg
of L-dihydroorotic acid, 0.031 mL (0.176 mmol)
diisopropylethylamine, 15.5 mg (0.115 mmol) of
1-hydroxybenzotriazole hydrate, 22 mg (0.115 mmol) EDC. After 1 h,
the slurry was treated with 1 mL of dimethylformamide. The mixture
was stirred for 16 h and then concentrated in vacuo. The residue
was taken up in ethyl acetate and washed with water and saturated
brine, dried over magnesium sulfate, filtered and concentrated in
vacuo. The residue was purified by thin layer preparative
chromatography using (1/2/17 v/v/v/30% ammonium
hydroxide/methanol/dichlomethane) eluent to provide 34.2 mg of the
title product. TLC: Rf=0.33, 1/2/17 v/v/v/30% ammonium
hydroxide/methanol/dichlomethane).
[0427] HPLC: Rt=11.3 min; (.sup.1H)-NMR (CDCl.sub.3) consistent
with structure.
EXAMPLE 41
[0428] Compound 41.
[0429] To a solution of 42.8 mg of the resultant compound of
Example 40A in 5 mL dichloromethane was added sequentially, at
ambient temperature under an atmosphere of nitrogen, 17.2 mg of
N-tert-butyl glyoxalic acid, 0.032 mL diisopropylethylamine, 16 mg
of 1-hydroxybenzotriazole hydrate, 22.6 mg EDC. The mixture was
stirred for 16 h and then concentrated in vacuo. The residue was
taken up in ethyl acetate and washed with water, 0.5 N hydrochloric
acid, washed with sodium bicarbonate, saturated brine, dried over
magnesium sulfate, filtered and concentrated in vacuo. The residue
was purified by thin layer preparative chromatography using 40%
ethyl acetate/dichloromethane eluent to provide 14.9 mg of the
title product. TLC: Rf=0.47, 40% ethyl acetate/dichloromethane,
HPLC: Rt=15.2 min; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 42
[0430] Compound 42.
[0431] To a solution of 43.5 mg of the resultant crude compound of
Example 40A in 5 mL dichloromethane was added sequentially at
ambient temperature, under an atmosphere of nitrogen, 13.0 mg of
succinamic acid, 0.024 mL diisopropylethylamine, 15.0 mg of
1-hydroxybenzotriazole hydrate, and 21.3 mg EDC. The mixture was
stirred for 16 h and then concentrated in vacuo. The residue was
taken up in ethyl acetate and washed with sodium bicarbonate,
saturated brine, dried over magnesium sulfate, filtered and
concentrated in vacuo. The residue was purified by thin layer
preparative chromatography using (1/2/11 v/v/v/30% ammonium
hydroxide/methanol/dichlomethane) eluent to provide 35.3 mg of the
title product. TLC: Rf=0.25, 1/2/11 v/v/v/30% ammonium
hydroxide/methanol/dichl- omethane, HPLC: Rt=11.6 min;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 43
[0432] Compound 43.
[0433] To a solution of 42.8 mg of the resultant compound of
Example 40A in 5 mL dichloromethane was added sequentially, at
ambient temperature under an atmosphere of nitrogen, with 14.1 mg
of L-pyroglutamic acid, 0.024 mL diisopropylethylamine, 14.8 mg of
1-hydroxybenzotriazole hydrate, 20.9 mg EDC. The mixture was
stirred for 16 h and then concentrated in vacuo. The residue was
taken up in ethyl acetate and washed with water, 0.5 N hydrochloric
acid, washed with sodium bicarbonate, saturated brine, dried over
magnesium sulfate, filtered and concentrated in vacuo. The residue
was purified by thin layer preparative chromatography using (1/2/11
v/v/v/30% ammonium hydroxide/methanol/dichlo- methane) eluent to
provide 29.9 mg of the title product. TLC: Rf=0.33, 1/2/11
v/v/v/30% ammonium hydroxide/methanol/dichlomethane, HPLC: Rt=11.7
min; (.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 44
[0434] A. 3-Pyridylmethyl-N-hydroxysucchinimdyl Carbonate.
[0435] To a solution of 181.0 mg of 3 pyidinecarbinol in 5 mL
acetonitrile was added sequentially at ambient temperature under an
atmosphere of nitrogen, with 0.72 mL diisopropylethylamine and
354.1 mg of N,N'-disuccinimidyl carbonate. After 4 h, the resultant
mixture was concentrated in vacuo to provide a yellow solid which
was used without subsequent purification.
[0436] B. Compound 44.
[0437] To a solution of 58.1 mg of the resultant crude compound of
Example 40A in 3 mL of dichloromethane was added sequentially, at
ambient temperature under an atmosphere of nitrogen, 0.075 mL
diisopropylethylamine and 46.3 mg of the resultant compound of
Example 20A. The mixture was stirred for 16 h and then concentrated
in vacuo. The residue was taken up in diethyl ether and extracted
into 3.times.25 mL of 0.5N HCl. The combined aqueous extracts were
adjusted to pH 8 with solid sodium bicarbonate and extracted into
3.times.25 mL ethyl acetate. The combined organic extracts were
washed with saturated brine, dried over magnesium sulfate,
filtered, and concentrated in vacuo. The residue was purified by
thin layer preparative chromatography using (1/2/17/20 v/v/v/30%
ammonium hydroxide/methanol/dichlomethane/diethyl ether) eluent to
provide 10.3 mg of the title product. TLC: Rf=0.4, 1/2/17/20
v/v/v/30% ammonium hydroxide/methanol/dichlomethane/diethyl ether,
HPLC: Rt=11.8 min; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 45
[0438] Compound 45.
[0439] To a solution of 28.3 mg of the resultant compound of
Example 39A in 4 mL of dichloromethane was added 1 mL saturated
aqueous sodium bicarbonate solution, 9.2 mg sodium bicarbonate, and
0.013 mL of benzenesulfonyl chloride. After 14 h, the resulting
mixture was diluted with ethyl acetate, washed with saturated
brine, dried over magnesium sulfate, filtered and concentrated in
vacuo. The residue was purified by thin layer preparative
chromatography using 10% diethyl ether/dichloromethane eluent to
provide 19.3 mg of the title product. TLC: Rf=0.84, 25% diethyl
ether/dichlormethane, HPLC: Rt=17.2 min; (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 46
[0440] Compound 46.
[0441] To a solution of 47.0 mg (0.140 mmol) of the resultant
compound of Example 39A in 4 mL of dichloromethane was added 1 mL
saturated aqueous sodium bicarbonate solution, 17.6 mg of solid
sodium bicarbonate, and 41.4 mg of 2,4 dimethylthiazole-5-sulfonyl
chloride. After 14 h, the resulting mixture was diluted with ethyl
acetate, washed with saturated brine, dried over magnesium sulfate,
filtered and concentrated in vacuo. The residue was purified by
thin layer preparative chromatography using 25% ethyl
acetate/dichloromethane eluent to provide 34.6 mg of the title
product. TLC: Rf=0.44, 25% diethyl ether/dichloromethane, HPLC:
Rt=16.4 min; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 47
[0442] Compound 47.
[0443] To a solution of 50.7 mg of the resultant compound of
Example 39A in 4 mL of dichloromethane was added 1 mL saturated
aqueous sodium bicarbonate solution, 15.2 mg of solid sodium
bicarbonate, and 2-fluorobenzenesulfonyl chloride 35.2 mg. After 14
h, the resulting mixture was diluted with ethyl acetate, washed
with saturated brine, dried over magnesium sulfate, filtered and
concentrated in vacuo. The residue was purified by thin layer
preparative chromatography using 10% diethyl ether/dichloromethane
eluent to provide 40.5 mg of the title product. TLC: Rf=0.44, 25%
diethyl ether/dichloromethane, HPLC: Rt=17.2 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 48
[0444] A. N-succinimidlyl-(S)-3-tetrahydrofuryl Carbonate.
[0445] To a solution of 12.5 mL of 1.93 M phosgene in toluene at
0-5.degree. C. was added 1.3 g of
(S)-(+)-3-hydroxy-tetrahydrofuran. After stirring for 2 h, the
reaction mixture was sparged with nitrogen and then concentrated to
dryness in vacuo to provide 1.486 g of crude chloroformate. This
material was taken up in 10 mL of acetonitrile and treated
sequentially at ambient temperature under an atmosphere of nitrogen
with 1.17 g of N-hydroxysuccinimide and 1.41 mL of triethylamine.
After stirring for 14 h, the reaction mixture was concentrated in
vacuo to provide 3.44 g of the title product as a white solid.
[0446] B. Compound 48.
[0447] To a solution of 87.2 mg of the resultant compound of
Example 40A in 5 mL of dichloromethane was added sequentially, at
ambient temperature under an atmosphere of nitrogen, 0.113 mL
diisopropylethylamine and 68 mg of the resultant compound of
Example 48A. The mixture was stirred for 16 h and then concentrated
in vacuo. The residue was taken up in ethyl acetate and washed with
water, 0.5 N HCl, saturated sodium bicarbonate, saturated brine,
dried over magnesium sulfate, filtered, and concentrated in vacuo.
The residue was purified by silica gel chromatography using
(3/6/20/65 v/v/v/v/30% ammonium hydroxide/methanol/diethyl
ether/dichlomethane) eluent followed by crystallization from a
mixture of dichloromethane, diethyl ether, and hexanes to provide
58 mg of the title product. TLC: Rf=0.17, 75k ethyl
acetate/dichloromethane, HPLC: Rt=13.1 min.; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 49
[0448] Compound 49.
[0449] Following the procedure described in Example 83, a solution
of the resultant compound of Example 39A in CH.sub.2Cl.sub.2 is
reacted with 2,4-difluorobenzenesulfonyl chloride in the presence
of water and NaHCO.sub.3. Following dilution with additional
CH.sub.2Cl.sub.2 and aqueous workup, the resultant product is dried
over MgSO.sub.4 filtered, and concentrated in vacuo. The residue is
then purified by silica gel chromatography using an appropriate
solvent system to yield the title product.
EXAMPLE 50
[0450] Compound 50.
[0451] A solution of 30 mg of the resulting compound of Example 58
and 9 .mu.L of dimethysulfamoyl chloride in 10 mL of
CH.sub.2Cl.sub.2 was reacted in the same manner as described for
Example 14. After workup and purification by preparative
reversed-phase C.sub.18HPLC using a linear gradient of 35% to 100%
CH.sub.3CN/H.sub.2O with 0.1% TFA as eluent, 6.5 mg of the title
compound was obtained. TLC: Rf=0.2, 3% CH.sub.3OH in
CH.sub.2Cl.sub.2, HPLC: Rt=15.96 min; (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 51
[0452] A. Compound XXI (A=Tert-butoxycarbonyl, D'=Isobutyl,
A'=Benzyloxycarbonyl).
[0453] To a solution of the resultant compound of Example 39A
(2.5g, 7.43 mmol) in CH.sub.2Cl.sub.2 (50 mL) was added
triethylamine (2.1 mL, 14.9 mmol) followed by addition of benzyl
chloroformate (1.2 mL, 8.1 mmol). The mixture was allowed to stir
at ambient temperature for 6 h. The solution was diluted with 1 L
of CH.sub.2Cl.sub.2 and washed with water. The organics were dried
over anhydrous MgSO.sub.4, concentrated under reduced pressure,
then purified via silica gel chromatography. Gradient solvent
system: CH.sub.2Cl.sub.2 followed by 3:97 methanol/CH.sub.2Cl.sub-
.2. The title compound (2.97 g, was obtained as a colorless oil.
TLC: Rf=0.14, 3:97 methanol/CH.sub.2Cl.sub.2; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
[0454] B. Compound XXI (A=H, D'=Isobutyl, A'=Benzyloxycarbonyl,
Hydrochloride Salt).
[0455] To a solution of 1.5 g (3.187 mmol) of the resultant
compound of Example 51A in ethyl acetate (25 mL) at -20.degree. C.
was bubbled anhydrous HCl gas for 10 min. The ice bath was removed
and after an additional 15 min. the reaction mixture was sparged
with nitrogen, then concentrated in vacuo to provide 1.29 g of
title product as a white solid which was used directly for ensuing
reaction. TLC: Rf=0.14, 10% methanol/CH.sub.2Cl.sub.2.
[0456] C. Compound XXI (A=(S)-3-tetrahydrofuryloxycarbonyl,
D'=Isobutyl, A'=Benzyloxycarbonyl).
[0457] To a solution of 1.077 g of the resultant crude compound of
Example 51B (2.647 mmol) in acetonitrile (10 mL) was added
sequentially at ambient temperature under an atmosphere of
nitrogen, 1.61 mL (9.263 mmol) of diisopropylethylamine and 910 mg
(3.97 mmol) of the resultant compound of Example 48A. After
stirring for 3 h, an additional 223 mg (0.973 mmol) of the
resultant compound of Example 48A was added. The mixture was
stirred for 16 h and then concentrated in vacuo. The residue was
taken up in ethyl acetate and washed with water, 0.5 N HCl,
saturated sodium bicarbonate, saturated brine, dried over magnesium
sulfate, filtered, and concentrated in vacuo. The residue was
purified by low pressure silica gel column chromatography using a
gradient 10% to 25% ethyl acetate in CH.sub.2Cl.sub.2 eluent to
yield 1.025 g of the title product as a white solid. TLC: Rf=0.10,
10% ethyl acetate/CH.sub.2Cl.sub.2; (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
[0458] D. Compound XXI (A=(S)-3-tetrahydrofuryloxycarbonyl,
D'=Isobutyl, A'=H).
[0459] A solution of 872 mg (1.799 mmol) of the resultant compounds
of Example 51C in (10 mL) of ethyl alcohol was added, at ambient
temperature under a nitrogen atmosphere, to a slurry of 87 mg (10%
by weight) of 10% palladium on carbon in (5 mL) ethyl alcohol and
hydrogenated for 16 h under a slight positive pressure of hydrogen.
The mixture was filtered and concentrated in vacuo to yield 553.2
mg of the title product as a colorless glass which was used
directly for ensuing reaction. TLC: Rf=0.46, 10%
methanol/CH.sub.2Cl.sub.2.
[0460] E. Compound 51.
[0461] To a solution of 72.7 mg (0.207 mmol) of the resultant
compound of Example 51D in CH.sub.2Cl.sub.2 (4 mL) was added
aqueous sodium bicarbonate (1 mL), solid sodium bicarbonate 22.6 mg
(0.27 mmol), and 2-(pyrid-2-yl)-thiophene-5-sulfonyl chloride 64.6
mg, (0.249 mmol). After 14 h, the resulting mixture was diluted
with ethyl acetate, washed with saturated brine, dried over
magnesium sulfate, filtered and concentrated in vacuo. The residue
was purified by thin layer preparative chromatography using 15 to
30% ethyl acetate/CH.sub.2Cl.sub.2 eluent to provide 53 mg of the
title product as a white solid. TLC; RF =0.25, 25% ethyl
acetate/CH.sub.2Cl.sub.2, HPLC: Rt=15.3 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 52
[0462] A. N-hydroxysuccinimidyl-(RS)-3-hydroxyl-tetrahydrofuryl
Carbonate.
[0463] The title compound was prepared as described in Example 48A
starting with 1.0 g of (RS)-3-hydroxy-tetrahydrofuran and yielding
2.33 g of a white solid.
[0464] B. Compound 52.
[0465] To a solution of 105 mg of the resultant compound of Example
35A in CH.sub.2Cl.sub.2 was added, at ambient temperature under an
atmosphere of nitrogen, 112 mg of the resultant compound of Example
52A and 126 mg N,N-diisopropylethylamine. The mixture was stirred
for 4 h, diluted with CH.sub.2Cl.sub.2 washed and saturated
NaHCO.sub.3 and saturated NaCl, then dried over MgSO.sub.4,
filtered, and concentrated in vacuo. The residue was purified by
low pressure silica gel chromatography using 5% CH.sub.3OH in
CH.sub.2Cl.sub.2 as eluent to yield 101.4 mg of product. TLC:
Rf=0.52, 5% CH.sub.3OH in CH.sub.2Cl.sub.2, HPLC: Rt=15.05 min.
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 53
[0466] Compound 53.
[0467] To a solution of 72.3 mg (0.19 mmol) of the resultant
compound of Example 51D in CH.sub.2Cl.sub.2 (4 mL) was added
aqueous sodium bicarbonate (1 mL), solid sodium bicarbonate 19.2 mg
(0.228 mmol), and 4-acetamido-3-chlorobenzene sulfonyl chloride
61.1 mg, (0.228 mmol). After 14 h, the resulting mixture was
diluted with EtOAc, washed with saturated brine, dried over
magnesium sulfate, filtered and concentrated in vacuo. The residue
was purified by low pressure silica gel column chromatography using
20% to 45% EtOAc/CH.sub.2Cl.sub.2 eluent to provide 49.1 mg of the
title product. TLC: RF=0.29, 50% EtOAc/CH.sub.2Cl.sub.2, HPLC:
Rt=13.9 min; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 54
[0468] Compound 54.
[0469] A solution of 260 mg of the resulting Compound of 39A and 45
mg of 3-acetamido-4-fluorobenzenesulfonyl chloride in 10 mL of
CH.sub.2Cl.sub.2 was reacted in the same manner as described for
Example 14. After workup and purification by preparative
reversed-phase C.sub.18HPLC using a linear gradient of 35% to 100%
CH.sub.3CN/H.sub.2O with 0.1% TFA as eluent, 1.4 mg of the title
compound was obtained. TLC: Rf=0.25, 5% CH.sub.3OH in
CH.sub.2Cl.sub.2, HPLC: Rt=15.63 min; (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 55
[0470] Compound 55.
[0471] 35.0 mg of the resulting compound of Example 54 was treated
with 1 mL of 90% aqueous TFA and allowed to stand for 12 h. The
mixture was concentrated in vacuo and the residue taken up in 10 mL
of dry CH.sub.2Cl.sub.2, treated with 34 .mu.L of DIEA (0.23
mmoles) and 20 mg of 1-benzyl-3-tert-butyl-1H-pyrazole-5-carbonyl
chloride. The mixture was stirred for 1.5 h, then diluted with in
CH.sub.2Cl.sub.2, and washed with 1 N HCl. After drying over
MgSO.sub.4 and concentrating in vacua, a portion of the mixture was
purified by preparative reversed-phase C.sub.18HPLC using a linear
gradient of 35% to 100% CH.sub.3CN/H.sub.2O with 0.1% TFA for
elution to obtain 1.1 mg of the title compound. TLC: Rf=0.8, 5%
CH.sub.3OH in CH.sub.2Cl.sub.2, HPLC: Rt=18.25 min; 1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 56
[0472] A. S(-)-1-phenylethyl-N-hydroxysuccinimdyl Carbonate.
[0473] The title compound was prepared from 9.5 .mu.L of
S(-)-1-phenylethanol and 30 m of N,N-disuccinimidyl carbonate as
described in Example 44A. The resulting material was used without
subsequent purification; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
[0474] B. Compound 56.
[0475] 45.0 mg of the resulting compound of Example 58 was treated
with 1 mL of 90% aqueous TFA and allowed to stand for 12 h. The
mixture was concentrated in vacuo and the residue taken up in 15 mL
of dry CH.sub.2Cl.sub.2, treated with the above mixed anhydride and
65 .mu.L of triethylamine. The mixture was stirred for 14 h then
diluted with ethyl acetate and washed with saturated sodium
bicarbonate solution and saturated brine, dried over magnesium
sulfate, filtered and concentrated in vacuo. A portion of the
mixture was purified by preparative reversed-phase C.sub.18HPLC
using a linear gradient of mixture was purified by preparative
reversed-phase C.sub.18HPLC using a linear gradient of 35% of 100%
CH.sub.3CN/H.sub.2O with 0.1% TFA for elution to obtain 1.1 mg of
the title compound. TLC: Rf=0.5, 3% CH.sub.3OH in CH.sub.2Cl.sub.2,
HPLC: Rt=17.44 min; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 57
[0476] Compound 57.
[0477] 30 mg of the resultant compound of Example 58 was treated
with 1 mL of 90% aqueous TFA and allowed to stand for 12 h. The
mixture was concentrated in vacuo and the residue taken up in 25 mL
of dry CH.sub.2Cl.sub.2, washed and saturated sodium bicarbonate
solution, dried over magnesium sulfate, filtered and concentrated
in vacuo. A solution of 14 mg of the resultant free amine in 10 mL
of CH.sub.2Cl.sub.2 was treated with 6 .mu.L of phenoxyacetyl
chloride and 12 .mu.L of triethylamine. The mixture was stirred
under an inert atmosphere for 1 h, then diluted in CH.sub.2Cl.sub.2
and washed with 1 N HCl. After drying over MgSO.sub.4 and
concentrating in vacuo. A portion of the mixture was purified by
preparative reversed-phase C.sub.18HPLC using a linear gradient of
35% to 100% CH.sub.3CN/H.sub.2O with 0.1% TFA as eluant to obtain
16.5 mg of the title compound. TLC: Rf=0.25, 3% MeOH in
CH.sub.2Cl.sub.2, HPLC: Rt=16.6 min; (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 58
[0478] Compound 58.
[0479] A solution of 500 mg of the resulting compound of Example
39A and 370 mg of benzofurazan-4-sulfonyl chloride in 10 mL of
CH.sub.2Cl.sub.2 was reacted in the same manner as described for
Example 14. After workup, the title compound was obtained by
crystallization from hot ethanol. Further purification of this
material by preparative reversed-phase C.sub.18HPLC using a linear
gradient of 35% to 100% CH.sub.3CN/H.sub.2O with 0.1% TFA as eluent
gave 2.0 mg of the title compound. TLC: Rf=0.35, 3% CH.sub.3OH in
CH.sub.2Cl.sub.2, HPLC: Rt=17.00 min; (1H)-NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 59
[0480] A. R(+)-1-phenylethyl-N-hydroxysuccinimdyl Carbonate.
[0481] The title compound was prepared from R(+)-1-phenylethanol as
described in Example 56A to yield a white solid. The resulting
material was used directly for subsequent reaction; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
[0482] B. Compound 59.
[0483] A 36 mg portion of the resultant compound of Example 58 and
0.21 .mu.mol of the resulting compound of 59A were reacted in the
manner described in example 56B. After workup and purification by
preparative reversed-phase C.sub.18HPLC using a linear gradient of
35% to 100% CH.sub.3CN/H.sub.2O with 0.1% TFA as eluant, 1.0 mg of
the title compound was obtained as a white solid. TLC: Rf=0.45, 3%
MeOH in CH.sub.2Cl.sub.2, HPLC: Rt=17.34 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 60
[0484] Compound 60.
[0485] To a solution of 70 mg of the resultant compound of Example
51D in 10 mL of CH.sub.2Cl.sub.2 was added 3 mL of saturated
aqueous sodium bicarbonate solution, 50 mg of sodium bicarbonate,
and 53 mg of benzofurazan-4-sulfonyl chloride. The mixture was
stirred vigorously for 4 h, then the resulting mixture was diluted
with CH.sub.2Cl.sub.2, washed with saturated brine, dried over
magnesium sulfate and filtered. After concentration of the mixture
in vacuo, the residue was purified by thick layer silica gel
chromatography using 5% MeOH/CH.sub.2Cl.sub.2 as eluant to obtain
80 mg of the title compound as a white solid. TLC: Rf=0.80, 5% MeOH
in CH.sub.2Cl.sub.2, HPLC: Rt=14.96 min; (.sup.1H)-NMR (CD.sub.3)
consistent with structure.
EXAMPLE 61
[0486] Compound 61.
[0487] To a solution of 35.5 mg (0.076 mmol) of the resultant
compound of Example 16 in 1 mL of dichloromethane was sequentially
added 27.6 .mu.L (0.159 mmol) of diisopropylethyl amine and 12
.mu.L (0.083 mmol) of benzyl chloroformate. After 1 hour, the
mixture was concentrated in vacuo. The residue was purified by
preparative thin layer chromatography with 50% ethyl
acetate/dichloromethane as an eluent to yield 38.7 mg of the title
compound as a white solid; TLC: Rf=0.63, 50% ethyl
acetate/dichlormethane; HPLC: Rt=15.45 min; (1H)-NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 62
[0488] A. Benzofurazan-4-sulfonic Acid.
[0489] To a solution of 252.0 mg (1.05 mmol) of
o-nitroaniline-m-sulfonic acid sodium salt in 1 mL of water was
added 0.52 mL of 2.0 N HC1. After 1/2 h, 0.68 mL (1.05 mmol) of
tertrabutylammonium hydroxide (40% in water) was added. After 2
hours, the mixture was concentrated in vacua. A solution of the
residue in 7 mL of acetic acid was treated with 488.5 mg (1.10
mmol) of lead tertraacetate. After 24 hours, the precipitate was
filtered and washed with small amount of acetic acid. The solid was
further dried in vacuo to yield 267.9 mg of product. TLC: Rf=0.09,
10% CH.sub.3OH/CH.sub.2Cl.sub.2.
[0490] B. Benzofurazan-4-sulfonyl Chloride.
[0491] To a solution of 137.0 mg (0.522 mmol) of triphenylphosphine
in 0.5 mL of dichloromethane was slowly added 47 .mu.L (0.594 mmol)
of sulfuric chloride at 0C. The ice-water bath was removed and the
crude resultant compound of Example 62A in 0.5 mL of
dichloromethane was added slowly. After 3 hours, the mixture was
treated with 30 mL of 50% ether/hexane. The supernatant was
decanted into a dry flask and concentrated in vacuo. The residue
was purified by filtering through a plug of silica gel with 25%
ethyl acetate as an eluent to yield 23 mg of product. TLC: Rf=0.6,
10% CH.sub.3OH/CH.sub.2Cl.sub.2; (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
[0492] C. Compound 62.
[0493] To a solution of 55.7 mg (0.166 mmol) of the resultant
compound of Example 39A in 1 mL of dichloromethane was sequentially
added 0.5 mL of saturated NaHCO.sub.3, a small amount of solid
NaHCO.sub.3 and the resultant compound of Example 62B. After 3
hours, the mixture was diluted with dichloromethane. The two layers
were separated and the aqueous layer was extracted once with
dichloromethane. The combined organic layer was washed with brine
then dried over MgSO.sub.4, filtered and concentrated in vacuo. The
residue was purified by preparative HPLC to yield 5.3 mg of the
title compound as a white solid; TLC: Rf=0.40, 50% ethyl
acetate/dichloromethane; HPLC Rt=16.5 min; H)-NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 63
[0494] A.
[0495] A Solution of 3.0 mg (0.0058 mmol) of the resultant title
compound of Example 62 in 2 mL ethyl acetate was treated with HCl
gas (moderate stream) for 3 minutes. The mixture was concentrated
in vacuo to yield the crude amine hydrochloride salt. TLC: Rf=0.20,
10% CH.sub.3OH/CH.sub.2Cl.s- ub.2.
[0496] B. Compound 63.
[0497] To a solution of the crude resultant compound of Example 63A
in 1 mL of dichloromethane was sequentially added 2.1 uL (0.0121
mmol) of diisopropyl ethyl amine and 0.9 uL (0.0064 mmol) of benzyl
chloroformate. After 1 hour, the mixture was concentrated in vacuo.
The residue was purified by preparative thin layer chromatography
with 90% dichloromethane/methanol as an eluent to yield 2.6 mg of
the title compound as a white solid; TLC: Rf=0.34, 50% ethyl
acetate/dichloromethane; HPLC, Rt=17.1 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 64
[0498] A. 5-(Dimethylamino)thioxomethoxy)-benzofurazan.
[0499] To a solution of 500 mg (3.67 mmol) of 5-hydroxybezofurazan
in 10 mL of DMF was added 140 mg (4.59 mmol) of NaH in small
portions. The resulting mixture was stirred at room temperature
until no more gas evolved. The flask was then immersed in a cold
water bath and 540 mg (4.41 mmol) of dimethylthiocarbamoyl chloride
(from Aldrich) was added. After 5 minutes, the water bath was
removed the mixture was heated to 80.degree. C. for 1 hour. After
being cooled to room temperature, the mixture was poured into 20 mL
of 0.5 N NaOH three times and water three times. The solid was
dried in vacuum to yield 580 mg of product that was used in the
next reaction without further purification; TLC: Rf=0.20, 20% ethyl
acetate/hexane; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
[0500] B. 5-((Dimethylamino)carbonyl)thio)-benzofurazan.
[0501] The crude product, 510 mg (2.28 mmol), from Example 64A was
heated to 190.degree. C. in a sealed tube. After 5 hours, it was
cooled to room temperature and ethyl acetate was added. The
solution was filtered through a plug of a silica and concentrated
in vacuo to yield 360 mg of product which was again used in the
next reaction without further purification. TLC: Rf=0.20, 20% ethyl
acetate/hexane.
[0502] C. 5-Mercaptobenzofurazan.
[0503] To a solution of 357.4 mg (1.60 mmol) of the resultant
compound of Example 64B in 2 mL of methanol was added 7 mL of 6 N
NaOH. The mixture was heated to 90.degree. C. for 2 hours. The
mixture was poured into 100 mL ice and acidified with concentrated
HCl. The slurry was filtered and rinsed three times with water. The
residue was dried in vacuo to yield 145.6 mg of product; TLC:
Rf=0.70, 20 ethyl acetate/hexane; (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
[0504] D. Benzofurazan-5-sulfonyl Chloride.
[0505] Chlorine gas was bubbled through a solution of 39.9 mg (0.26
mmol) of the resultant compound of Example 64C in a mixture of 1 mL
of ethyl acetate and 0.5 mL of water for 3 minutes. The mixture was
then washed repeatedly with brine until no more precipitate formed.
The organic layer was dried over MgSO.sub.4, filtered and
concentrated to yield 30 mg of the product (52%). TLC: Rf=0.22, 20%
ethyl acetate/hexane.
[0506] E. Compound 64.
[0507] A solution of the resultant compounds of Examples 52D and
39A (total yields) in a mixture of 1 mL of dichloromethane, 0.3 mL
of saturated NaHCO.sub.3 and a small amount of solid NaHCO.sub.3
was stirred at room temperature for 2 hours. The solution was
diluted with 30 mL of dichloromethane and the two layers were
separated. The aqueous layer was extracted once with
dichloromethane chloride. The combined organic layer was washed
with brine, dried over MgSO.sub.4 and concentrated. The residue was
purified by preparative thin layer chromatography with 90%
dichloromethane/ether as an eluent to yield 30 mg of the title
product as a white solid; TLC: Rf=0.46, 10%
Et.sub.2O/CH.sub.2Cl.sub.2, HPLC Rt=17.6 min; (.sup.1H)-NMR
(CDCl.sub.3): .delta. 8.45 (s), 1H; 7.96(d), 1H; 7.65 (d), 1H;
7.25(m), 5H; 4.65(d), 1H; 3.85(m), 1H; 3.78(m), 1H; 3.30(d), 2H;
3.10(m), 2H; 2.90(m), 2H: 1.90(m), 1H; 1.40(s), 9H; 0.90 (d),
6H.
EXAMPLE 65
[0508] Compound 65.
[0509] A solution of 13.1 mg (0.025 mmol) of the resultant compound
of Example 64E in 1.5 mL of ethyl acetate was treated with gaseous
HCl (moderate stream) at 0.degree. C. for 3 minutes. The solvent
was removed to yield a solid residue which was used in the next
reaction without further purification; TLC: Rf=0.52, 10%
CH.sub.3OH/CH.sub.2Cl.sub.2. A solution of this hydrochloride salt
(entire yield) in 1 mL of dichloromethane was treated sequentially
with 9.2 .mu.L (0.053 mmol) of diisopropyl ethyl amine and 4.0
.mu.L (0.028 mmol) of benzyl chloroformate. After 3 hours, the
mixture was concentrated and purified by preparative thin layer
chromatography with 90% dichloromethane/ether as an eluent to yield
11.7 mg of the title compound as a white solid; TLC: Rf=0.65, 10%
Et.sub.2O/CH.sub.2Cl.sub.2; HPLC Rt=17.6 min; (.sup.1H)-NMR
(CDCl.sub.3: .delta. 8.45(s), 1H; 7.96(d), 1H; 7.65(d), 1H;
7.25(m), 1OH; 5.00,(m), 2H; 4.85(d), 1H; 3.86(m), 2H; 3.60(bs), 1H;
3.25(m), 12H; 3.05(d), 2H; 2.96(m), 1H: 2.98(m), 1H; 1.88(m), 1H;
0.90(dd), 6H.
EXAMPLE 66
[0510] Compound 66.
[0511] A solution of 100 mg (0.46 mmol) of the resultant compound
of Example 64D and 101 mg (0.286 mmol) of the resultant compound of
Example 48A in a mixture of 2 mL of dichloromethane, 0.5 mL of
saturated NaHCO.sub.3 and small amount of solid NaHCO.sub.3 was
stirred at room temperature for 2 hours. The solution was diluted
with 50 mL of dichloromethane and the two layers were separated.
The aqueous layer was extracted once with dichloromethane. The
combined organic layer was washed with brine, dried over MgSO.sub.4
and concentrated. The residue was purified by preparative thin
layer chromatography with 20% ethyl acetate/hexane as an eluent to
yield 82 mg of the title product as a slightly impure pale yellow
solid. The material was further purified by preparative HPLC with a
linear gradient solvent system of 35% to 80% of acetonitrile/water
(0.1% TFA) over 80 min. Upon removal the solvents 50 mg of white
solid was obtained. TLC: Rf=0.46, 10% Et.sub.2O/CH.sub.2Cl.su- b.2;
HPLC, Rt=17.6 min; (.sup.1H)-NMR (CDCl.sub.3): .delta. 8.45 (s),
1H; 7.96 (d), 1H; 7.65 (d), 1H; 7.25 (m), 5H; 5.15 (m), 1H: 4.85
(d), 1H; 3.82 (m) 4H; 3.68 (d), 1H; 3.20(m), 2H, 3.05 (d), 2H; 2.96
(m), 1H; 2.88 (m), 1H; 2.14(m), 1H; 1.92(m), 2H; 1.50(bs), 1H;
0.90(dd), 6H.
EXAMPLE 67
[0512] Compound 67.
[0513] Following the procedure described in Example 40B, a solution
of the resultant compound of Example 40A in CH.sub.2Cl.sub.2 is
treated with bis- ((carboxamido)-amino)-acetic acid,
diisopropylethylamine, HOBt, and EDC in a 1:1:1:1:1 molar ratio.
the mixture is stirred for 16 h at ambient temperature while
protected from moisture, then diluted with additional
CH.sub.2Cl.sub.2 and washed sequentially with H.sub.2O, saturated
NaHCO.sub.3 solution and brine, then dried over MgSO.sub.4 and
concentrated in vacuo. The residue is purified by silica gel
chromatography using an appropriate eluant to yield the title
product.
EXAMPLE 68
[0514] Compound 68. This compound was prepared by the route
described in Example 26, except that the reacting amine used was
the resulting compound of Example 39A (146 mg, 0.43 mmol) and the
acylating agent was 4-fluorophenyl sulphonyl chloride (27 mg, 0.14
mmol). After chromatographic purification on a silica gel column
using 8% CH.sub.3OH/CH.sub.2Cl.sub.2 as eluent, 92.8 mg of the
title compound was obtained. HPLC: Rt=15.9 minutes. TLC: Rf=0.54,
8% MeOH/CH.sub.2Cl.sub.2; (.sup.1H)-NMR (CDCl.sub.3) consistent
with structure.
EXAMPLE 69
[0515] A.
[0516] The resulting compound of Example 68 (72.1 mg, 0.167 mmol)
was dissolved in 90% aqueous TFA (3.3 mL), and the reaction mixture
stirred for 3 hours at room temperature, then was concentrated to
dryness. TLC: Rf=0.29, 8% MeOH/CH.sub.2Cl.sub.2.
[0517] B. Compound 69.
[0518] To a solution of the resulting compound of Example 69A (41.7
mg, 0.09 mmol) in CH.sub.2Cl.sub.2 (2 mL) was added
diisopropylethtylamine (47 .mu.l, 0.27 mmol) and the resulting
compound of Example 48A (33 mg, 0.15 mmol), and the reaction
proceeded for 14 hours at room temperature. The reaction mixture
was then concentrated, and the residue was chromatographed on a
silica gel column using 8% THF/CH.sub.2Cl.sub.2 was eluent,
yielding the desired compound which was further subjected to
purification by preparative HPLC, yielding 7.8 mg of a white solid.
HPLC: Rt=13.5 minutes. TLC: Rf=0.36, 8-6 THF/CH.sub.2Cl.sub.2;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 70
[0519] Compound 70.
[0520] A solution of 30 mg of the resulting compound of Example 54
and 17.6 mg of 3-acetamido-4-fluorobenzenesulfonyl chloride in 10
mL of CH.sub.2Cl.sub.2 was reacted in the same manner as described
for Example 14. After workup and purification by preparative
reversed-phase C.sub.16HPLC using a linear gradient of 35% to 100%
CH.sub.3CN/H.sub.2O with 0.1% TFA as eluent, 2.0 mg of the title
compound was obtained. TLC: Rf=0.5, 10% CH.sub.3OH in
CH.sub.2Cl.sub.2, HPLC: Rt=13.74 min; (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 71
[0521] Compound 71.
[0522] A 30 mg portion of the resultant compound of Example 58 was
deprotected with trifluoroacetic acid and the resulting compound
reacted with 9 .mu.L of dimethysulfamoyl chloride in 10 mL of
CH.sub.2Cl.sub.2 was reacted in the manner described in Example 14.
After workup and purification by preparative reversed-phase
C.sub.18HPLC using a linear gradient of 35% to 100%
CH.sub.3CN/H.sub.2O with 0.1% TFA as eluant, 6.5 mg of the title
compound was obtained. TLC: Rf=0.2, 3% MeOH in CH.sub.2Cl.sub.2,
HPLC: Rt=15.96 min; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 72
[0523] Compound 72.
[0524] A solution of the resulting compound from the
trifluoroacetic acid deprotection of Example 69A (31 mg, 0.07 mmol)
in CH.sub.2Cl.sub.2 (2 mL) was added diisopropylethylamine (47
.mu.l, 0.27 mmol) and dimethylsulfamoyl chloride (22 .mu.l, 0.20
mmol), and the reaction proceeded for 16 hours at room temperature.
The reaction mixture was then concentrated, and the residue was
chromatographed on a thick layer silica gel plate (1.0 mm) using 5%
THF/CH.sub.2Cl.sub.2 as eluent, yielding the desired compound which
was further subjected to purification by preparative HPLC to yield
7.8 mg of a white solid. HPLC: Rt=14.8 minutes. TLC: Rf=0.44, 5%
THF/CH.sub.2Cl.sub.2 Example 73
[0525] Compound 73.
[0526] A 43 mg portion of the resultant compound of Example 54 was
treated with 1 mL of 90% aqueous TFA and allowed to stand for 12 h.
The mixture was concentrated in vacuo and the residue taken up in 5
mL of CH.sub.2Cl.sub.2. To this solution, 3 mL saturated aqueous
sodium bicarbonate and 25 mg of 2,5-dimethoxybenzenesulfonyl
chloride was added, and the mixture was stirred for 12 h, warming
slowly to ambient temperature. After concentration of the mixture
in vacuo, the residue was purified by thick layer silica gel
chromatography using 3% MeOH/CH.sub.2Cl.sub.2 as eluant followed by
preparative reversed-phase C.sub.18HPLC using a linear gradient of
35% to 100% CH.sub.3CN/H.sub.2O with 0.1% TFA as eluant to obtain
5.5 mg of the title compound. TLC: Rf=0.20, 3%
MeOH/CH.sub.2Cl.sub.2, HPLC: Rt=15.15 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 74
[0527] A. Compound XXI (A=Tert-butoxycarbonyl,
D'=Cyclopropylmethyl, A'=H).
[0528] To a solution of compound XX (A=tert-butoxycarbonyl) (0.8 g,
2.67 mmol) in ethanol (30 mL) was added a solution of KOH (0.18 g,
3.2 mmol) in ethanol (20 mL) and the mixture stirred for 45 min at
room temperature. In a separate flask, a solution of
cyclopropylmethyl-amine hydrochloride (1.44 g, 13.3 mmol) in
ethanol (20 mL) was added KOH (0.75 g, 13.3 mmol). The mixture was
stirred 30 min at room temperature. The solutions were combined and
heated at 85.degree. C. for 3 h. The solution was concentrated
under reduced pressure and the residue slurried in diethyl ether
and filtered. The ethereal layer was concentrated to give 0.32 g of
a white solid; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
[0529] B. Compound 74.
[0530] To a solution of the resulting compound of Example 74A (0.1
g, 0.30 mmol) in CH.sub.2Cl.sub.2 (20 mL) was added a saturated
solution of sodium bicarbonate, followed by addition of solid
sodium bicarbonate (30 mg, 0.36 mmol), then 4-fluorobenzenesulfonyl
chloride (0.07 g, 0.36 mmol). The mixture was allowed to stir at
room temperature for 4 h. The organics were extracted into 250 mL
CH.sub.2Cl.sub.2, dried over anhydrous MgSO.sub.4, concentrated
under reduced pressure then purified via medium pressure liquid
chromatography using a gradient system of CH.sub.2Cl.sub.2 followed
by 0.5:99.5 methanol/CH.sub.2Cl.sub.2 followed by 1:99
methanol/CH.sub.2Cl.sub.2. The title compound was obtained as 35 mg
of a colorless foam. HPLC: Rt=16.8 min. TLC: Rf=0.32, 3:97
methanol/CH.sub.2Cl.sub.2; (.sup.1H)-NMR (CDCl.sub.3) consistent
with structure.
EXAMPLE 75
[0531] A. Compound XXI (A=Tert-butoxycarbonyl, D'=Isopropyl,
A'=H).
[0532] To a solution of Compound XX (A=tert-butoxycarbonyl) (1.67
mmol) in ethanol (10 mL) was treated with isopropylamine (10 mL).
The solution was heated to 85.degree. C. for 72 h. The solution was
filtered then concentrated under reduced pressure to give 0.56 g of
the title compound which was used without subsequent purification.
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
[0533] B. Compound 75.
[0534] To a solution of the resultant compound of Example 75A (0.2
g, 0.65 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added a saturated
solution of sodium bicarbonate (3 mL), followed by addition of
solid sodium bicarbonate (0.11 g, 1.31 mmol), then
p-fluorobenzenesulfonyl chloride (0.25 g, 1.28 mmol). The mixture
was stirred overnight at ambient temperature. The organics were
extracted into 100 mL CH.sub.2Cl.sub.2, dried over anhydrous
MgSO.sub.4, concentrated under reduced pressure then purified via
medium pressure silica gel chromatography using a gradient system
of CH.sub.2Cl.sub.2 followed by 1:99 methanol/CH.sub.2Cl.sub.2. The
title compound was obtained as a colorless foam 200 mg. TLC:
Rf=0.22, 3:97 methanol/CH.sub.2Cl.sub.2, HPLC: Rt=16.48 min;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 76
[0535] A. Compound XXI (A=Tert-butoxycarbonyl, D'=Morpholinyl,
A'=H).
[0536] To a solution of compound XX (A=Boc) in ethanol is added 3
molar equivalents of N-amino morpholine. The mixture is heated
under reflux for 12 h, cooled, and the mixture concentrated in
vacuo. The residue is purified by preparative reversed-phase
chromatography using a linear gradient of 5% to 100%
acetonitrile/H.sub.2O as eluant to yield the title compound.
[0537] B. Compound 76.
[0538] Following the procedure described in Example 81, a solution
of the resultant compound of Example 76A in CH.sub.2Cl.sub.2 is
reacted with 4-fluorobenzenesulfonyl chloride in the presence of
water and NaHCO.sub.3. Following dilution with additional
CH.sub.2Cl.sub.2 and aqueous workup, the resultant product is dried
over MgSO.sub.4, filtered, and concentrated in vacuo. The residue
is then purified by silica gel chromatography using an appropriate
solvent system to yield the title product.
EXAMPLE 77
[0539] A. Compound XXI (A=Tert-butoxycarbonyl,
D'=4-(N,N-dimethylamino)-be- nzyl, A'=H).
[0540] To a solution of compound XX (A=Boc) in ethanol is added 3
molar equivalents of 4-aminomethyl-(N,N-dimethyl)-aniline. The
mixture is heated under reflux for 12 h, cooled, and the mixture
concentrated in vacuo. The residue is purified by silica gel
chromatography using an appropriate solvent system as eluant to
yield the title product.
[0541] B. Compound 77.
[0542] Following the procedure described in Example 81, a solution
of the resultant compound of Example 77A in CH.sub.2Cl.sub.2 is
reacted with 4-fluorobenzenesulfonyl chloride in the presence of
water and NaHCO.sub.3. Following dilution with additional
CH.sub.2Cl.sub.2 and aqueous workup, the resultant product is dried
over MgSO.sub.4, filtered, and concentrated in vacuo. The residue
is then purified by silica gel chromatography using an appropriate
solvent system to yield the title product.
EXAMPLE 78
[0543] A. Compound XXI (A=Tert-butoxycarbonyl, D'=Cyclopentyl,
A=H).
[0544] To a solution of compound XX (A=Boc) in ethanol is added 10
molar equivalents of cyclopentylamine. The mixture is heated under
reflux for 12 h, cooled, and the mixture concentrated in vacuo. The
residue is used without subsequent purification.
[0545] B. Compound 78.
[0546] Following the procedure described in Example 81, a solution
of the resultant compound of Example 78A in CH.sub.2Cl.sub.2 is
reacted with 4-fluorobenzenesulfonyl chloride in the presence of
water and NaHCO.sub.3. Following dilution with additional
CH.sub.2Cl.sub.2 and aqueous workup, the resultant product is dried
over MgSO.sub.4, filtered, and concentrated in vacuo. The residue
is then purified by silica gel chromatography using an appropriate
solvent system to yield the title product.
EXAMPLE 79
[0547] A. Compound XXI (A=Tert-butoxycarbonyl,
D'=2-(4-pyridyl)ethyl, A'=H).
[0548] To a solution of compound XX (A=Boc) in ethanol is added 3
molar equivalents of 4-aminoethylpyridine. The mixture is heated
under reflux for 12 h, cooled, and the mixture concentrated in
vacuo. The residue is purified by preparative reversed-phase
chromatography using a linear gradient of 5% to 100%
acetonitrile/H.sub.2O as eluant to yield the title product.
[0549] B. Compound 79.
[0550] Following the procedure described in Example 81, a solution
of the resultant compound of Example 79A in CH.sub.2Cl.sub.2 is
reacted with 4-fluorobenzenesulfonyl chloride in the presence of
water and NaHCO.sub.3. Following dilution with additional
CH.sub.2Cl.sub.2 and aqueous workup, the resultant product is dried
over MgSO.sub.4, filtered, and concentrated in vacuo. The residue
is then purified by silica gel chromatography using an appropriate
solvent system to yield the title product.
EXAMPLE 80
[0551] A. 4-Cyanotetrahydro-4H-pyran.
[0552] Following essentially the procedure of Yoneda, R.
"Cyanophosphate: An Efficient intermediate for Conversion of
Carbonyl compounds to Nitriles," Tetrahedron Lett., 30, 3681
(1989), a solution of tetrahydro-4H-pyran-one (9.9 g, 97.8 mmol) in
dry THF (50 mL) is reacted with lithium cyanide (9.7 g, 294 mmol)
and diethylcyanophosphonate (24 g, 146 mmol). The mixture is
stirred for 24 h at ambient temperature. The reaction is quenched
by the addition of 100 mL H.sub.2O. The product is extracted into
1.5 L of diethyl ether, dried over anhydrous MgSO.sub.4 then
concentrated under reduced pressure. The residue is dissolved in
dry THF (30 mL) and tert-butyl alcohol (7.25 g, 97.8 mmol). This
solution is added slowly to 75 mL of a 1 M solution of SmI.sub.2.
The mixture is stirred for 15 h at ambient temperature. The
reaction is quenched by addition of 100 mL of saturated aqueous
NH.sub.4Cl. The resulting mixture is extracted with diethyl ether
and the organic layers dried over anhydrous MgSO.sub.4 and
concentrated under reduced pressure. Purification by silica gel
chromatography gives the title compound.
[0553] B. 4-(aminomethyl)tetrahydro-4H-pyran
[0554] To a solution of the compound of the Example 80A (10 g, 89.9
mmol) in absolute ethanol (200 mL) is added Raney Nickel (2.0 g,
50% slurry in water). The mixture is stirred for 24 hours at
ambient temperature under 40 psig of hydrogen. The solution is
filtered through celite and the solution concentrated under reduced
pressure. The residue is taken up in ether (2L) washed with brine,
dried in anh. MgSO.sub.4, then concentrated under reduced pressure
to give the title compound.
[0555] C.
(1S,2R)-N-(1-Benzyl-3-(N-(4-(aminomethyl)tetrahydro-4H-pyran))-2-
-hydroxypropyl)-tert butoxycarbonylamine.
[0556] To a solution of the compound of Example 80B (5g, 48.5 mmol)
in absolute ethanol (20 mL) is added the compound XX (A=Boc)(2.55
g, 9.7 mmol). The mixture is stirred for 24 hours at ambient
temperature. The solution is concentrated under reduced pressure
and the crude product is puffed via column chromatography to give
the title compound.
[0557] D. Compound XXII (A=Boc, D'=(4-tetrahydro-4H-pyranyl)methyl,
A'=H).
[0558] To a solution of compound XX (A=Boc) in ethanol is added 3
molar equivalents of the resulting compound of Example 80C. The
mixture is heated under reflux for 12 h, cooled, and the mixture
concentrated in vacuo. The residue is purified by preparative
reversed-phase chromatography using a linear gradient of 5% to 100%
acetonitrile/H.sub.2O as eluant to yield the title compound.
[0559] To a solution of compound XX(A=Boc) in ethanol is added 3
molar equivalents of N-amino morpholine. The mixture is heated
under reflux for 12 h, cooled, and the mixture concentrated in
vacuo. The residue is purified by preparative reversed-phase
chromatography using a linear gradient of 5% to 100%
acetonitrile/H.sub.2O as eluant to yield the title compound.
[0560] E. Compound 80.
[0561] Following the procedure described in Example 81, a solution
of the resultant compound of Example 80D in CH.sub.2Cl.sub.2 is
reacted with 4-fluorobenzenesulfonyl chloride in the presence of
water and NaHCO.sub.3. Following dilution with additional
CH.sub.2Cl.sub.2 and aqueous workup, the resultant product is dried
over MgSO.sub.4, filtered, and concentrated in vacuo. The residue
is then purified by silica gel chromatography to yield the title
product.
EXAMPLE 81
[0562] A. Compound XXII (A=Tert-butoxycarbonyl, D'=Isobutyl,
E=3,4-Dichlorophenyl).
[0563] A solution of 316 mg of the resultant compound of Example
39A in 4:1 CH.sub.2Cl.sub.2/saturated aqueous NaHCO.sub.3 was
treated sequentially, at ambient temperature under an atmosphere of
nitrogen, with 276 mg of 3,4-dichorobenzenesulfonyl chloride and 95
mg of sodium bicarbonate. The mixture was stirred for 14 h, diluted
with CH.sub.2Cl.sub.2, washed with saturated NaCl then dried over
MgSO.sub.4, filtered, and concentrated in vacuo. The residue was
purified by low pressure silica gel chromatography using 5% diethyl
ether/CH.sub.2Cl.sub.2 as, eluent to yield 490 mg of product. TLC:
Rf=0.26, 5% diethyl ether in CH.sub.2Cl.sub.2, HPLC: Rt=18.92 min.
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
[0564] B. Compound XXII (A=H, D'=isobutyl, E=3,4-dichlorophenyl,
Hydrochloride Salt).
[0565] A solution of 467 mg of the resultant compound of Example
81A in ethyl acetate was treated at -20.degree. C. with HCl gas.
The HCl was bubbled through the mixture for 20 min over which time
the temperature was allowed to warm to 20.degree. C. Nitrogen was
then bubbled through the mixture for 15 min and solvent removed in
vacuo to yield 412 mg of product as a white solid which was used
without subsequent purification.
[0566] C. Compound 81.
[0567] A solution of 91 mg of the resultant compound of Example 81B
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 25 mg of allyl
chloroformate and 52 mg N,N-diisopropylethylamine. The mixture was
stirred for 4 h and then concentrated in vacuo. The residue was
taken up in ethyl acetate and washed with 0.5 N HCl and saturated
NaCl then dried over MgSO.sub.4, filtered, and concentrated in
vacuo to yield 89 mg of the title product as a white solid. TLC:
Rf=0.53, 5% diethyl ether in CH.sub.2Cl.sub.2, HPLC: Rt=17.95 min.
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 82
[0568] A. (3-Pyridyl)-methyl-4-nitrophenyl-carbonate.
[0569] To a solution of 3.65 g of bis-(nitrophenyl) carbonate in 25
mL of CH.sub.2Cl.sub.2 at 0.degree. C. was added sequentially 0.97
mL of 3-pyridyl carbinol and 1.3 mL of 4-methyl morphine. After
stirring at room temperature for 24 hours, the resultant mixture
was diluted with 100 mL of CH.sub.2Cl.sub.2, washed with saturated
sodium bicarbonate, water and brine, dried over magnesium sulfate,
filtered and concentrated in vacuo. The residue was purified by
filtration through a plug of silica gel, using 0-40%
EtOAc/CH.sub.2Cl.sub.2 as eluent to provide 1.68 g of the title
product. TLC: Rf=0.19, 50% EtOAc/hexane.
[0570] B. Compound XXII (A=Tert-butoxycarbonyl, D'=Isobutyl,
E=3,4-Benzofurazan).
[0571] To a solution of 498.6 mg of the resultant compound of
Example 39A in 10 mL of CH.sub.2Cl.sub.2 was added sequentially, 2
mL of saturated sodium bicarbonate, a small amount of solid sodium
bicarbonate and 518.4 mg of the resultant compound of Example 64D.
After stirring at room temperature for 3 hours, the resultant
mixture was diluted with 60 mL of CH.sub.2Cl.sub.2, washed with
saturated sodium bicarbonate and brine, dried over magnesium
sulfate, filtered and concentrated in vacuo. The residue was
purified by silica gel chromatography using 5% diethyl ether/hexane
as eluent to yield 300 mg of white solid. TLC: Rf=0.80, 50%
EtOAc/hexane.
[0572] C. Compound XXII (A=H, D'=Isobutyl; E=3,4-Benzofurazan,
Hydrochloride Salt.).
[0573] A solution of 60.3 mg of the resultant compound of Example
82B in 3 mL of EtOAc at -20.degree. C. was treated with anhydrous
HCl gas for 5 min. The ice bath was removed and after an additional
10 min. The reaction mixture was sparged with nitrogen then
concentrated in vacuum and the resulting white solid used without
subsequent purification for subsequent reaction.
[0574] D. Compound 82.
[0575] To a solution of the resultant compound of Example 82C
(entire yield) in 2 mL of CH.sub.2Cl.sub.2 was added sequentially,
45 .mu.L of diisopropylethylamine and 35.1 mg of the resultant
compound of Example 82A. The mixture was stirred for 24 hours and
then concentrated in vacuo. The residue was purified by preparative
thin layer chromatography using 60% ether/CH.sub.2Cl.sub.2 as
eluent followed by preparative reversed-phase C.sub.18HPLC using a
linear gradient of 40% to 100% CH.sub.3CN/H.sub.2O with 0.1% TFA as
eluant. The resultant TFA salt of the title compound was washed
with saturated sodium bicarbonate to yield 6.5 mg of the title
compound. TLC: Rf=0.15, 20% EtOAc/CH.sub.2Cl.sub.2, HPLC: Rt=13.52
min. (.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 83
[0576] A. Compound XXII (A=Tert-butoxycaronyl, D'=Isobutyl,
E=4-acetamido-3-chlorophenyl).
[0577] A solution of 339 mg of the resultant compound of Example
39A in 4:1 CH.sub.2Cl.sub.2/saturated aqueous NaHCO.sub.3 was
treated sequentially, at ambient temperature under an atmosphere of
nitrogen, with 324 mg of 4-acetamido-3-chlorobenzenesulfonyl
chloride and 102 mg of sodium bicarbonate. The mixture was stirred
for 14 h, diluted with CH.sub.2Cl.sub.21 washed with saturated NaCl
then dried over MgSO.sub.4, filtered, and concentrated in vacuo.
The residue was purified by low pressure silica gel chromatography
using 20% diethyl ether in CH.sub.2Cl.sub.2 as eluent to yield 498
mg of product. TLC: Rf=0.27 (20% diethyl ether in
CH.sub.2Cl.sub.2). HPLC: Rt=16.20 min. (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
[0578] B. Compound XXII (A=H, D'=Isobutyl,
E=4-acetamido-3-chlorophenyl, Hydrochloride Salt).
[0579] A solution of 474 mg of the resultant compound of Example
83A in ethyl acetate was treated at -20.degree. C. with HCl gas.
The HCl was bubbled through the mixture for 20 min over which time
the temperature was allowed to warm to 20.degree. C. Nitrogen was
then bubbled through the mixture for 15 min and the solvent was
removed in vacuo to yield 421 mg of product as a white solid which
was used without subsequent purification.
[0580] C. Compound 83.
[0581] A solution of 92 mg of the resultant compound of Example 83B
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 24 mg of allyl
chloroformate and 52 mg N,N-diisopropylethylamine. The mixture was
stirred for 4 h and then concentrated in vacuo. The residue was
taken up in ethyl acetate and washed with 0.5 N HCl and saturated
NaCl, then dried over MgSO.sub.4, filtered, and concentrated in
vacuo to yield 106 mg of the title product as a white solid. TLC:
Rf=0.38 (20% diethyl ether in CH.sub.2Cl.sub.2). HPLC: Rt=15.28
min. .sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 84
[0582] Compound XXII (A=Tert-butoxycarbonyl, D'=Isobutyl,
E=3,4-dichlorophenyl).
[0583] To a solution of the resultant compound of Example 51D (220
mg, 0.61 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added the
3,4-dichlorobenzenesulfonyl chloride (300 mg, 1.22 mmol) followed
by the addition of a saturated solution of sodium bicarbonate (3
mL) followed by addition of 0.1 g of solid sodium bicarbonate. The
mixture was stirred at ambient temperature overnight. The solution
was diluted with 100 mL CH.sub.2Cl.sub.2, the organics separated,
dried over anhydrous, MgSO.sub.4, and the organics concentrated
under reduced pressure to obtain 0.17 g of crude product. The crude
product was purified via medium pressure liquid chromatography
using CH.sub.2Cl.sub.2 followed by 0.5:99.5
methanol/CH.sub.2Cl.sub.2 followed by a 1:99
methanol/CH.sub.2Cl.sub.2 solution as the solvent system to give
103 mg of the title compound as a white solid. TLC: Rf=0.56 (3:97
methanol/CH.sub.2Cl.sub.2), HPLC: Rt=19.78 min, (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 85
[0584] A. (3-Tetrahydrofuryl)-methyl-4-nitrophenyl-carbonate.
[0585] To a solution of 1.21 g of p-nitrophenyl chloroformate in 20
mL of CH.sub.2Cl.sub.2 0.degree. C. was added sequentially, 0.51 g
of tetrahydro-3-furanmethanol and 0.66 mL of 4-methyl morpholine.
After stirring at room temperature for 2 hours. The mixture was
stirred for 2 hours and concentrated in vacuo. The residue was
purified by filtering through a plug of silica gel, using 0-50%
EtOAc/CH.sub.2Cl.sub.2 as eluent to provide 1.17 g of the title
product as a pale yellow solid. TLC: Rf=0.20, 50% EtOAc/hexane.
[0586] B. Compound 85.
[0587] To a solution of 70 mg of the resultant compound of Example
81B in 1 mL of THF was added sequentially, 56 .mu.L of
diisopropylethylamine and a solution of 46.6 mg of the resultant
compound of Example 85A in 1 mL of THF. The mixture was stirred for
24 hours and then concentrated in vacuo. The residue was diluted
with 60 mL of CH.sub.2Cl.sub.2, washed with 5% sodium bicarbonate
and brine, dried over magnesium sulfate, filtered and concentrated
in vacuo to yield 120 mg of crude product. The residue was purified
by preparative thin layer chromatography using 20%
EtOAc/CH.sub.2Cl.sub.2 as eluent to yield 82 mg of the title
compound. TLC: Rf=0.4, 20% EtOAc/CH.sub.2Cl.sub.2, HPLC: Rt=17.08
min. .sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 86
[0588] Compound 86.
[0589] A solution of 42 mg of the resultant compound of Example 40A
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 41 mg of the
product of Example 52A and 46 mg N,N-diisopropylethylamine. The
mixture was stirred 14 h, diluted with CH.sub.2Cl.sub.2, washed
with saturated NaHCO.sub.3 and saturated NaCl, then dried over
MgSO.sub.4, filtered, and concentrated in vacuo. The residue was
purified by preparative thin layer silica gel chromatography using
ethyl acetate as eluent to yield 43 mg of product. TLC: Rf=0.44
(20% ethyl acetate). HPLC: Rt=13.14 min. (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 87
[0590] A. Compound XXII (A=H, D'=Isobutyl, E=4-acetamido,
3-fluoro).
[0591] A solution of 25 mg of the resultant compound of Example 54
in EtOAc (10 mL) at 0.degree. C. was treated with anhydrous
hydrogen chloride gas for 10 min., and allowed to stand for 12 h
while warming to ambient temperature. The resultant mixture was
then concentrated in vacuo to yield compound as a white solid which
was used without subsequent purification for ensuing reaction.
[0592] B. Compound 87.
[0593] A 0.045 mmol portion of the resultant compound of Example
87A was taken up in 5 mL of CH.sub.2Cl.sub.2. To this solution, 40
.mu.L of diisopropylethylamine and 6 .mu.L of allyl chloroformate
were added at 0.degree. C. and the mixture was stirred for 12 h,
while warming slowly to ambient temperature. The resulting mixture
was diluted with CH.sub.2Cl.sub.2, washed with saturated brine,
dried over magnesium sulfate and filtered. After concentrated in
vacuo, the residue was purified by preparative reversed-phase
C.sub.18HPLC using a linear gradient of 35% to 100%
CH.sub.3CN/H.sub.2O with 0.1% TFA as eluant to obtain 11.6 mg of
the title compound. TLC: Rf=0.20, 5% MeOH/CH.sub.2Cl.sub.2, HPLC:
Rt=14.6 min; H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 88
[0594] Compound 88.
[0595] A 0.033 mmol portion of the resultant compound of Example
87A was taken up in 5 mL of CH.sub.2Cl.sub.2. To this solution, 26
.mu.L of triethylamine and 12 mg of the resultant compound of
Example 48A were added and stirred for 12 h. The resulting mixture
was diluted with CH.sub.2Cl.sub.2, washed with saturated sodium
bicarbonate solution and saturated brine, dried over magnesium
sulfate and filtered. After concentration of the mixture in vacuo,
the residue was purified by thick layer silica gel chromatography
using 5% MeOH/CH.sub.2Cl.sub.2 as eluant followed by preparative
reversed-phase C.sub.18HPLC using a linear gradient of 35% to 100%
CH.sub.3CN/H.sub.2O with 0.1% TFA as eluant to obtain 7.5 mg of the
title compound. TLC: Rf=0.30, 5% MeOH/CH.sub.2Cl.sub.2, HPLC:
Rt=13.38 min; .sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 89
[0596] Compound 89.
[0597] A solution of 28 mg of the resultant compound of Example 81B
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 8 mg of n-propyl
chloroformate and 17 mg N,N-diisopropylethylamine. The mixture was
stirred for 3 h and then concentrated in vacuo. The residue was
taken up in ethyl acetate and washed with 0.5 N HCl and saturated
NaCl then dried over MgSO.sub.4, filtered, and concentrated in
vacuo to yield 31 mg of the title product as a white solid. TLC:
Rf=0.35 (5% diethyl ether in CH.sub.2Cl.sub.2). HPLC: Rt=18.12 min.
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 90
[0598] Compound 90.
[0599] A solution of 28 mg of the resultant compound of Example 83B
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 7 mg of n-propyl
chloroformate and 15 mg N,N-diisopropylethylamine. The mixture was
stirred for 3 h and then concentrated in vacuo. The residue was
taken up in ethyl acetate and washed with 0.5 N HCl and saturated
NaCl then dried over MgSO.sub.4, filtered, and concentrated in
vacuo to yield 30 mg of the title product as a white solid. TLC:
Rf=0.47 (20% diethyl ether in CH.sub.2Cl.sub.2). HPLC: Rt=15.41
min. (.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 91
[0600] A. 3-Acetamidobenzene Sulfonic Acid.
[0601] A solution of 1.48 g of 3-aminobenzene sulfonic acid in 1:1
tetrahydrofuran/water was treated at 0.degree. C. with 1.43 g of
sodium bicarbonate. After 5 min, 1.30 g of acetic anhydride was
added dropwise and the reaction allowed to warm to ambient
temperature under an atmosphere of nitrogen over 14 h. The reaction
mixture was passed through a column of Amberlyst 15 ion exchange
resin, eluted with water, and concentrated in vacuo to yield an oil
which upon treatment with benzene and azeotropic removal of water
in vacuo yielded 1.8 g of the title product as a white crystalline
solid. (.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
[0602] B. 3-Acetamidobenzene Sulfonic Acid, Sodium Salt.
[0603] The resultant compound of Example 91A in water was treated
at 0.degree. C. with 8.5 mL of 1 N sodium hydroxide. The mixture
was stirred for 3 h and concentrated in vacuo to yield an oil which
upon treatment with benzene and azeotropic removal of water in
vacuo yielded the title product as a tan solid which was used
directly in the next reaction.
[0604] C. 3-Acetamidobenzenesulfonyl Chloride.
[0605] The resultant compound of Example 91B in CH.sub.2Cl.sub.2
was treated at 0.degree. C. with 4.5 g of phosphorous pentachloride
under an atmosphere of nitrogen. The mixture was stirred 14 h,
extracted with CH.sub.2Cl.sub.2, and concentrated in vacuo to yield
1.7 g of the title product as a brown oil. TLC: Rf=0.21 (1:1
toluene/diethyl ether). (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
[0606] D. Compound XXII (A=Tert-butoxycarbonyl, D'=Isobutyl,
E=3-acetamidophenyl).
[0607] A solution of 280 mg of the resultant compound of Example
39A in 4:1 CH.sub.2Cl.sub.2/saturated aqueous NaHCO.sub.3 was
treated sequentially, at ambient temperature under an atmosphere of
nitrogen, with 252 mg of the resultant compound of Example 91C and
105 mg of sodium bicarbonate. The mixture was stirred for 60 h,
diluted with CH.sub.2Cl.sub.2, washed with saturated NaCl then
dried over MgSO.sub.4, filtered and concentrated in vacuo. The
residue was purifed by low pressure silica gel chromatography using
20% diethyl ether in CH.sub.2Cl.sub.2 as eluent to yield 156 mg of
the title product. TLC: Rf=0.14 (20% diethyl ether in
CH.sub.2Cl.sub.2was). HPLC: Rt=15.39 min. (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
[0608] E. Compound XXII (A=H, D'=Isobutyl, E=3-acetamidophenyl,
Hydrochloride Salt).
[0609] A solution of 123 mg of the resultant compound of Example
91D in ethyl acetate was treated at -20.degree. C. with HCl gas.
The HCl was bubbled through the mixture for 20 min, over which time
the temperature was allowed to warm to 20.degree. C. Nitrogen was
then bubbled through the mixture for 15 min and solvent removed in
vacuo to yield 118 mg of the title product as a white solid which
was used directly in subsequent reactions.
[0610] F. Compound 91.
[0611] A solution of 49 mg of the resultant compound of Example 91E
in CH.sub.2Cl.sub.2 was added, at ambient temperature under an
atmosphere of nitrogen, to a solution of 48 mg of the resultant
compound of Example 48A and 54 mg N,N-diisopropylethylamine in
CH.sub.2Cl.sub.2. The mixture was stirred for 14 h, diluted with
CH.sub.2Cl.sub.2, washed with saturated NaHCO.sub.3 and saturated
NaCl, then dried over MgSO.sub.4, filtered and concentrated in
vacuo. The residue was subjected to preparative thin layer silica
gel chromatography using 5% CH.sub.3OH in CH.sub.2Cl.sub.2 to yield
42 mg of product. TLC: Rf=0.32 (5% CH.sub.3OH in CH.sub.2Cl.sub.2).
HPLC: Rt=13.27 min. (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 92
[0612] Compound 92.
[0613] To a solution of 63.5 mg of the resultant compound of
Example 17B, diastereomer B in 1 mL of THF was added sequentially,
52 .mu.L of diisopropylethylamine and a solution of 43.3 mg of the
resultant compound of Example 85A in 1 mL of THF. The mixture was
stirred for 24 hours and then concentrated in vacuo. The residue
was diluted with 60 mL of CH.sub.2Cl.sub.2, washed with 5% sodium
bicarbonate and brine, dried over magnesium sulfate, filtered and
concentrated in vacuo to yield 70.7 mg of crude product. The
residue was purified by preparative reversed-phase C.sub.18 HPLC
using a linear gradient of 30% to 100% CH.sub.3CN/H.sub.2O with
0.1% TFA as eluant to obtain 43.9 mg of the title compound. TLC:
Rf=0.29, 100% EtOAc. HPLC: Rt=13.24 min; (.sup.1H) NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 93
[0614] A. N-hydroxysuccinimidyl-(R)-3-hydroxytetrahydrofuryl
Carbonate.
[0615] The title compound was prepared as described in Example 48A
starting with 81 mg of (R)-3-hydroxytetrahydrofuran to yield 56 mg
of the title product as a white solid. (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
[0616] B. Compound 93.
[0617] To a solution of 43 mg of the resultant compound of Example
35A in CH.sub.2Cl.sub.2 was added, at ambient temperature under an
atmosphere of nitrogen, 27 mg of the resultant compound of Example
93A and 39 mg N,N-diisopropylethylamine. The mixture was stirred
for 14 h, diluted with CH.sub.2Cl.sub.2, washed with saturated
NaHCO.sub.3 and saturated NaCl, then dried over MgSO.sub.4,
filtered, and concentrated in vacuo. The residue was purified by
preparative thin layer silica gel chromatography using 2%
CH.sub.3OH in CH.sub.2Cl.sub.2 as eluent to yield 45 mg of the
title product as a white solid. TLC: Rf=0.52 (5% CH.sub.3OH
CH.sub.2Cl.sub.2). HPLC: Rt=14.94 min. (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 94
[0618] Compound 94.
[0619] A solution of 47 mg of the resultant compound of Example 40A
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 28 mg of the
product of Example 93A and 39 mg N,N-diisopropylethylamine. The
mixture was stirred for 14 h, diluted with CH.sub.2Cl.sub.2, washed
with saturated NaHCO.sub.3 and saturated NaCl, then dried over
MgSO.sub.4, filtered, and concentrated in vacuo. The residue was
purified by preparative thin layer silica gel chromatography using
5% methanol in CH.sub.2Cl.sub.2 as eluent to yield 40 mg of the
title product as a white solid. TLC: Rf=0.38 (ethyl acetate). HPLC:
Rt=13.09 min. (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 95
[0620] Compound 95.
[0621] To a solution of 72.0 mg (0.189 mmol) of the resultant
compound of Example 51D in CH.sub.2Cl.sub.2 (4 mL) was added
aqueous sodium bicarbonate (1 mL), solid sodium bicarbonate 19.1 mg
(0.227 mmol), and 2,3-dichlorothiophenesulfonyl chloride 57.1 mg,
(0.227 mmol). After 14 h, the resulting mixture was diluted with
EtOAc, washed with saturated brine, dried over magnesium sulfate,
filtered and concentrated in vacuo. The residue was purified by low
pressure silica gel column chromatography using 5 to 12%
EtOAc/CH.sub.2Cl.sub.2 eluent to provide 49.1 mg of the title
product. TLC: Rf=0.62 25% EtOAc/CH.sub.2Cl.sub.2HPLC: Rt=17.3 min;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 96
[0622] A. (4-Acetamido)-phenylmethyl-4-nitrophenyl-carbonate.
[0623] To a solution of 242.8 mg of p-nitrophenyl chloroformate in
5 mL of acetonitrile at 0.degree. C. was added sequentially, 165.2
mg of 4-acetamidobenzyl alcohol and 0.13 mL of 4-methyl morpholine.
The mixture was stirred for 24 hours and concentrated in vacuo. The
residue was taken up in CH.sub.2Cl.sub.2 and washed with 5% sodium
bicarbonate and brine, dried over magnesium sulfate, filtered and
concentrated in vacuo to yield 320 mg of the title compound. TLC:
Rf=0.23, 50% EtOAc/hexane.
[0624] B. Compound 96.
[0625] To solution of the resultant compound of Example 40A in 1 mL
of THF was added sequentially, 56 .mu.L of diisopropylethylamine
and 63 mg of the resultant compound of Example 96A. The mixture was
stirred for 24 hours and then concentrated in vacuo. The residue
was purified by preparative thin layer chromatography using 10%
methanol/CH.sub.2Cl.sub.2 as eluent followed by preparative
reversed-phase C.sub.18HPLC using a linear gradient of 30% to 100%
CH.sub.3CN/H.sub.2O with 0.1% TFA as eluant to yield 50.2 mg of the
title compound. TLC: Rf=0.43, 10% methanol/CH.sub.2Cl.sub.2, HPLC:
Rt=13.54 min. (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 97
[0626] Compound 97.
[0627] To solution of 60 mg of the resultant compound of Example
35A in 1 mL of THF was added sequentially, 54 .mu.L of
diisopropylethylamine and a solution of 48.9 mg of the resultant
compound of Example 85A in 1 mL THF. The mixture was stirred for 24
hours and then concentrated in vacuo. The residue was diluted with
60 mL of CH.sub.2Cl.sub.2 washed with 5% sodium bicarbonate and
brine, dried over magnesium sulfate, filtered and concentrated in
vacuo. The residue was purified by preparative thin layer
chromatography using 20% EtOAc/CH.sub.2Cl.sub.2 as eluent to yield
46.9 mg of the title compound. TLC: Rf=0.31, 20%
EtOAc/CH.sub.2Cl.sub.2. HPLC: Rt=15.18 min. (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 98
[0628] Compound 98.
[0629] To a solution of 61.0 mg of the resultant compound of
Example 35A in 1 mL of THF was added sequentially, 49 .mu.L of
diisopropylethylamine and a solution of 44 mg of the resultant
compound of Example 82A in 1 mL THF. The mixture was stirred for 24
hours and then concentrated in vacuo. The residue was purified by
preparative thin layer chromatography using 5%
methanol/CH.sub.2Cl.sub.2 as eluent to yield 61.0 mg of a white
solid. TLC: Rf=0.19, 5 methanol/CH.sub.2Cl.sub.2. HPLC: Rt=13.28
min; 13.28 min. (1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 99
[0630] Compound 99.
[0631] A solution of 75 mg of the resultant compound of Example 51D
and 45 mg of 4-chlorobenzenesulfonyl chloride were reacted in the
manner described in Example 60. After workup and purification by
preparative reversed-phase C18HPLC using a linear gradient of 35%
to 100% CH.sub.3CN/H.sub.2O with 0.1w TFA as eluant, 24.6 mg of the
title compound was obtained. TLC: Rf=0.3, 4% MeOH/CH.sub.2Cl.sub.2,
HPLC: Rt=15.87 min; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 100
[0632] Compound 100.
[0633] A solution of 40 mg of the resultant compound of Example 51D
and 45 mg of 4-methoxybenzenesulfonyl chloride were reacted in the
manner described in Example 60. After workup and purification by
preparative reversed-phase C.sub.18HPLC using a linear gradient of
35% to 1006 CH.sub.3CN/H.sub.2O with 0.1% TFA as eluant, 21.4 mg of
the title compound was obtained as a white solid. TLC: Rf=0.2, 4%
MeOH/CH.sub.2Cl.sub.2, HPLC: Rt=14.85 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 101
[0634] Compound 101.
[0635] This compound was prepared from the resultant compound of
Example 128 by treatment with hydrogen chloride gas and subsequent
reaction with the resultant compound of Example 48A in the manner
described in Example 132. After workup and purification by
preparative reversed-phase C.sub.18HPLC using a linear gradient of
35% to 100% CH.sub.3CN/H.sub.2O with 0.1% TFA as eluant on a
portion of the crude mixture, 4.2 mg of the title compound was
obtained as a white solid. TLC: Rf=0.2, 4% MeOH/CH.sub.2Cl.sub.2,
HPLC: Rt=11.53 min; (1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 102
[0636] Compound 102.
[0637] A solution of 36 mg of the resultant compound of Example 40A
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 8 mg of methyl
chloroformate and 22 mg N,N-diisopropylethylamine. The mixture was
stirred for 3 h and then concentrated in vacuo. The residue was
taken up in ethyl acetate and washed with 0.5 N HCl and saturated
NaCl then dried over MgSO.sub.4, filtered, and concentrated in
vacuo. The residue was purified by low pressure silica gel
chromatography using 30% diethyl ether in CH.sub.2Cl.sub.2 as
eluent to provide 27 mg of the title product as a white solid. TLC:
Rf=0.10 (30% diethyl ether in CH.sub.2Cl.sub.2). HPLC: Rt=13.49
min. (.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 103
[0638] Compound 103.
[0639] A solution of 29 mg of the resultant compound of Example 81B
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 6 mg of methyl
chloroformate and 17 mg N,N-diisopropylethylamine. The mixture was
stirred for 3 h and then concentrated in vacuo. The residue was
taken up in ethyl acetate and washed with 0.5 N HCl and saturated
NaCl then dried over MgSO.sub.4, filtered, and concentrated in
vacuo. The residue was purified by low pressure silica gel
chromatography using 5% diethyl ether/CH.sub.2Cl.sub.2 as eluent to
provide 29 mg of the title product as a white solid. TLC: Rf=0.24
(5% diethyl ether in CH.sub.2Cl.sub.2). HPLC: Rt=17.07 min.
.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 104
[0640] Compound 104.
[0641] A solution of 31 mg of the resultant compound of Example 35A
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 8 mg of methyl
chloroformate and 21 mg N,N-diisopropylethylamine. The mixture was
stirred for 3 h and then concentrated in vacuo. The residue was
taken up in ethyl acetate and washed with 0.5 N HCl and saturated
NaCl then dried over MgSO.sub.4, filtered, and concentrated in
vacuo. The residue was purified by low pressure silica gel
chromatography using 5% diethyl ether/CH.sub.2Cl.sub.2 as eluent to
provide 24 mg of the title product as a white solid. TLC: Rf=0.23
(5% diethyl ether in CH.sub.2Cl.sub.2). HPLC: Rt=15.41 min.
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 105
[0642] A. N-hydroxysuccinimidyl Methallyl Carbonate.
[0643] To a solution of 2.9 mL of 1.93 M phosgene in toluene at
-10.degree. C. was added 857 mg of methallyl alcohol. The mixture
was stirred for 2 h at -10.degree. C. to produce a 1.9 M solution
of the title compound which was used directly in subsequent
reactions.
[0644] B. Compound 105.
[0645] A solution of 39 mg of the resultant compound of Example 40A
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 0.05 mL of the
resultant compound of Example 105A and 24 mg
N,N-diisopropylethylamine. The mixture was stirred for 3 h and then
concentrated in vacuo. The residue was taken up in ethyl acetate
and washed with 0.5 N HCl and saturated NaCl then dried over
MgSO.sub.4, filtered, and concentrated in vacuo. The residue was
purified by preparative thin layer silica gel chromatography using
ethyl acetate as eluent to yield 18 mg of the title product as a
white solid. TLC: Rf=0.67 (ethyl acetate). HPLC: Rt=14.97 min.
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 106
[0646] Compound 106.
[0647] A solution of 31 mg of the resultant compound of Example 81B
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 0.04 mL of the
resultant compound of Example 105A and 18 mg
N,N-diisopropylethylamine. The mixture was stirred for 3 h and then
concentrated in vacuo. The residue was taken up in ethyl acetate
and washed with 0.5 N HCl and saturated NaCl then dried over
MgSO.sub.4, filtered, and concentrated in vacuo. The residue was
purified by low pressure silica gel chromatography using 5% diethyl
ether/CH.sub.2Cl.sub.2 as eluent to provide 19 mg of the title
product as a white solid. TLC: Rf=0.34 (5% diethyl
ether/CH.sub.2Cl.sub.2). HPLC: Rt=18.24 min. (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 107
[0648] Compound 107.
[0649] A solution of 28 mg of the resultant compound of Example 35A
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 0.05 mL of the
resultant compound of Example 105A and 19 mg
N,N-diisopropylethylamine. The mixture was stirred for 3 h and then
concentrated in vacuo. The residue was taken up in ethyl acetate
and washed with 0.5 N HCl and saturated NaCl then dried over
MgSO.sub.4, filtered, and concentrated in vacuo. The residue was
purified by low pressure silica gel chromatography using S% diethyl
ether in CH.sub.2Cl.sub.2 as eluent to provide 18 mg of the title
product as a white solid. TLC: Rf=0.25 (5% diethyl ether in
CH.sub.2Cl.sub.2). HPLC: Rt=16.68 min. (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 108
[0650] Compound 108.
[0651] To a solution of 62.5 mg of 124B in 1 mL of THF was added
sequentially 56 .mu.L of diisopropylethylamine and a solution of
49.6 mg of the resultant compound of Example 82A in 1 mL THF. The
mixture was stirred for 24 hours and then concentrated in vacuo.
The residue was purified by preparative thin layer chromatography
using 50% EtOAc/CH.sub.2Cl.sub.2 as eluent followed by preparative
reversed-phase C18HPLC using a linear gradient of 30% to 100%
CH.sub.3CN/H.sub.2O with 0.1% TFA as eluant on a portion of the
crude mixture, 4.2 mg of the title compound was obtained as a white
solid. TLC: Rf=0.16, 10% methanol/CH.sub.2Cl.sub.2, HPLC: Rt=13.67
min. (.sup.1H) NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 109
[0652] A. (S)-4-Methoxycarbonyl-oxazlidin-2-one.
[0653] To a solution of 4.88 g of serine methyl ester hydrochloride
in 25 mL of water was added 6.94 g of potassium carbonate. The
mixture was cooled to 0.degree. C. and 19.5 mL of phosgene was
added dropwise. After stirring at 0.degree. C. for 3 hours, water
was removed to yield a white solid with was washed with copious of
CH.sub.2Cl.sub.2. The organic solution was then dried over
magnesium sulfate, filtered and concentrated to yield 3.26 g of the
title product as a clear oil. (.sup.1H) NMR (D.sub.2O):
.delta.=3.82 (s, 3H), 4.43 (dd, 1H), 4.53 (dd, lh), 4.67 (t, 1H),
6.29 (s, 1H).
[0654] B. (S)-4-Hydroxymethyl-oxazlidin-2-one.
[0655] To a solution of 3.26 g of the resultant compound of Example
109A in 20 mL of ethanol at 0.degree. C. was added 0.85 g of sodium
borohydride in small portions. The ice bath was removed and after
additional 3 hours, 20 mL of 2.0 N hydrogen chloride was added to
the mixture, which was then concentrated to yield an oil. The
residue was extracted with EtOAc and the organic solution was dried
over magnesium sulfate, filtered and concentrated to yield 2.50 g
of the title compound. (.sup.1H) NMR (CDCl.sub.3): .delta.=2.48 (s,
1H), 3.69 (dd, 1H), 4.08 (m, 1H), 4.31 (t, 1H), 4.57 (t, 1H).
[0656] C. 4-Nitrophenyl-((S)-4-oxazlidin-2-onyl)-methyl
Carbonate.
[0657] To a solution of 1.04 g of p-nitrophenyl chloroformate in 20
mL of CH.sub.2Cl.sub.2 at 0.degree. C. was added sequentially, 0.5
g of the resultant compound of Example 109B and 0.6 mL of 4-methyl
morpholine. The mixture was stirred for 2 hours at ambient
temperature and then concentrated in vacuo. The residue was
purified by low pressure silica gel column chromatography using 20%
EtOAc in CH.sub.2Cl.sub.2 eluent to yield 0.57 g of the title
compound. TLC: Rf=0.10, 50% EtOAc/hexane.
[0658] D. Compound 109.
[0659] To a solution of 60 mg of the resultant compound of Example
35A in 1 mL of THF was added sequentially, 56 .mu.L of
diisopropylethylamine and a solution of 51.1 mg of the resultant
compound of Example 109C in 1 mL acetonitrile. The mixture was
stirred for 24 hours and then concentrated in vacuo. The residue
was purified by preparative thin layer chromatography using 5%
methanol/CH.sub.2Cl.sub.2 as eluent to yield 60.4 mg of the title
compound. TLC: Rf=0.38, 5% methanol/CH.sub.2Cl.sub.2, HPLC:
Rt=14.11 min. (.sup.1H) NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 110
[0660] Compound 110.
[0661] To a solution of 60 mg of the resultant compound of Example
40A in 1 mL of acetonitrile was added sequentially, 51 .mu.L of
diisopropylethylamine and a solution of 46.8 mg of the resultant
compound of Example 109C in 1 mL acetonitrile. The mixture was
stirred for 48 hours and then concentrated in vacuo. The residue
was purified by preparative thin layer chromatography using 10%
methanol/CH.sub.2Cl.sub.2 eluent followed by preparative
reversed-phase C18HPLC using a linear gradient of 30% to 100%
CH.sub.3CN/H.sub.2O with 0.1% TFA as eluant to yield 16 mg of the
title compound. TLC: Rf=0.28, 50% EtOAc/CH.sub.2Cl.sub.2. HPLC:
Rt=12.47 min. (.sup.1H) NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 111
[0662] A solution of 0.067 mmol of the resultant compound of
Example 114D in 5 mL of tetrahydrofuran was added 20 .mu.L of
diisopropylethylamine followed dropwise by a solution of the
resultant compound of Example 82A in 5 mL of tetrahydrofuran during
one hour. The mixture was stirred 16 h and then concentrated in
vacuo. The crude residue was purified by thick layer silica gel
chromatography using 5% MeOH/CH.sub.2Cl.sub.2 as eluant to obtain
21.8 mg of the title compound. TLC: Rf=0.45, 5%
MeOH/CH.sub.2Cl.sub.2; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 112
[0663] A. Compound XXII (A=tert-butoxycarbonyl, D'=Isobutyl,
E=3-sulfonamidophenyl).
[0664] To a solution of 96.6 mg (0.287 mmol) of the resultant
compound of Example 39A in CH.sub.2Cl.sub.2 (4 mL) was added
aqueous sodium bicarbonate (1 mL), solid sodium bicarbonate 36.2 mg
(0.431 mmol), and m-benzene disulfonylchloride 86.9 mg, (1.08
mmol). After stirring for 1 h, 30% ammonium hydroxide (10 mL) was
added. After 14 h the resulting mixture was diluted with
CH.sub.2Cl.sub.2, washed with saturated brine, dried over magnesium
sulfate, filtered and concentrated in vacuo. The residue was
purified by flash chromatography using 0 to 10%
methanol/CH.sub.2Cl.sub.2 eluent to provide 49.3 mg of the title
product. (.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
[0665] B. Compound XXII (A=H, D'=Isobutyl, E=3-sulfonamidophenyl,
Hydrochloride Salt).
[0666] A solution of 49.3 mg (0.089 mmol) of the resultant compound
of Example 112A in EtOAc (10 mL) at -20.degree. C. was treated with
anhydrous HCl gas for 10 min. The ice bath was removed and after an
additional 15 min., the reaction mixture was sparged with nitrogen
then concentrated in vacuo to provide 53.1 mg of title product as
the HCl salt. (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
[0667] C. Compound 112.
[0668] To a solution of 53.1 mg of the resultant compound of
Example 112B (0.089 mmol) in CH.sub.2Cl.sub.2 (3 mL) was treated
sequentially at ambient temperature under an atmosphere of
nitrogen, with 0.031 mL (0.177 mmol) diisopropylethylamine and 24.3
mg (0.106 mmol) of the resultant compound of Example 48A. The
mixture was stirred 16 h and then concentrated in vacuo. The
residue was taken up in CH.sub.2Cl.sub.2 and washed with saturated
brine, dried over magnesium sulfate, filtered, and concentrated in
vacuo. The residue was purified by low pressure silica gel column
chromatography using a gradient 5% to 20% EtOAc in CH.sub.2Cl.sub.2
as eluent to yield 10.8 mg of the title product. TLC: RF =0.4 25%
EtOAc in CH.sub.2Cl.sub.2, HPLC: Rt=13.3 min; (1H) NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 113
[0669] A. 3-Furansulfonyl Chloride.
[0670] In flame dried glassware under a nitrogen atmosphere to a
solution of 428 mg (2.909 mmol) of 3 bromofuran in anhydrous
tetrahydrofuran at -78.degree. C. was added 2.0 mL n-butyl lithium
(3.2 mmol at 1.6 molar in hexane). After 45 minutes the resultant
solution was added via cannula to a 20.degree. C. solution of
sulfuryl chloride in diethyl ether (5 mL plus a 2 mL rinse). After
1 h, the reaction was quenched with 0.5 N hydrochloric acid and
extraced into diethyl ether. The ethereal extracts were washed with
saturated brine, dried over magnesium sulfate, filtered and
concentrated in vacuo to provide 158 mg of the title product.
(.sup.1H) NMR (CDCl.sub.3) consistent with structure.
[0671] B. Compound XXII (A=Tert-butoxycarbonyl, D'=Isobutyl,
E=3-furyl).
[0672] To a solution of 289.7 mg (0.861 mmol) of the resultant
compound of Example 39A in CH.sub.2Cl.sub.2 (8 mL) was added
aqueous sodium bicarbonate (2 mL), solid sodium bicarbonate 108 mg
(1,292 mmol), and the resultant product from Example 113A 157.8 mg,
(1.08 mmol). After stirring for 1 h 30% ammonium hydroxide (10 mL)
was added. After 14 h, the resulting mixture was diluted with
CH.sub.2Cl.sub.2, washed with saturated brine, dried over magnesium
sulfate, filtered and concentrated in vacuo. The residue was
purified by flash chromatography using 1% to 15%
EtOAc/CH.sub.2Cl.sub.2.
[0673] C. Compound XXII (A=H, D'=Isobutyl, E=3-furyl, Hydrochloride
Salt).
[0674] A solution of 217.3 mg (0.581 mmol) of the resultant
compound of Example 113B in EtOAc (15 mL) at -20.degree. C. was
treated with anhydrous HCl gas for 10 min. The ice bath was removed
and after an additional 15 min. the reaction mixture was sparged
with nitrogen then concentrated in vacuo to provide 228 mg of title
product as the HCl salt. TLC: Rf=0.52 10%
methanol/CH.sub.2Cl.sub.2.
[0675] D. Compound 113.
[0676] To a solution of 65.3 mg of the resultant compound of
Example 113C (0.162 mmol) in CH.sub.2Cl.sub.2 (3 mL) was treated
sequentially at ambient temperature under an atmosphere of
nitrogen, with 0.056 mL (0.324 mmol) diisopropylethylamine and 44.6
mg (0.194 mmol) of the resultant compound of Example 48A. The
mixture was stirred 16 h and then concentrated in vacuo. The
residue was taken up in CH.sub.2Cl.sub.2 and washed with saturated
brine, dried over magnesium sulfate, filtered, and concentrated in
vacuo. The residue was purified by low pressure silica gel column
chromatography using a gradient 3% to 20% EtOAc in CH.sub.2Cl.sub.2
eluent to yield 10.8 mg of the title product. TLC: Rf=0.6, 25%
EtOAc/CH.sub.2Cl.sub.2, HPLC: Rt=13.9 min; (.sup.1H) NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 114
[0677] A. Aminomethylcyclopentane.
[0678] To a solution of LiAlH.sub.4 (38 g, 1.0 mole) in diethyl
ether (2 L) was added cyclopentanecarbonitrile (73.2 g, 0.77 mol)
as a solution in 250 mL ether. The solution was stirred overnight
at ambient temperature and then quenched by addition of the
organics to 3 L of a saturated potassium, sodium tartrate solution.
The amine was extracted into 3 L of ether, dried over anhydrous
K.sub.2CO.sub.3 then concentrated by distillation to approximately
400 mL total volume. The crude product was purified via
distillation to give 58.2 g of the title compound as a colorless
oil. (.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
[0679] B. Compound XXI (A=Tert-butoxycarbonyl,
D'=Cyclopentylmethyl, A'=H).
[0680] To the resultant compound of Example 114A (20 g, 0.2 mol)
was added compound XX (A=Boc) (5.84 g) and the mixture was stirred
for 24 h at ambient temperature. The solution was concentrated by
distillation under reduced pressure. The residue was triturated
with hexane and the solid collected by suction filtration and
washed with hexane to give 7.08 g of a white solid which was used
without further purification. TLC: Rf=0.59 (1:10:90 concentrated
NH.sub.4OH/methanol/CH.sub.2Cl.sub.2), (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
[0681] C. Compound XXII (A=Tert-butoxycarbonyl,
D'=Cyclopentylmethyl, E=4-fluorophenyl).
[0682] To a solution of the resultant compound of Example 114B (200
mg, 0.55 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added
4-fluorobenzenesulfonyl chloride (210 mg, 1.1 mmol) followed by the
addition of a saturated solution of sodium bicarbonate (3 mL)
followed by addition of solid sodium bicarbonate (0.1 g, 1.2 mmol).
The mixture was allowed to stir at ambient temperature overnight.
The solution was diluted with 100 mL CH.sub.2Cl.sub.2, the organics
separated, dried over anhydrous MgSO.sub.4, and the organics
concentrated under reduced pressure to obtain 0.33 g of crude
product. This material was purified via medium pressure liquid
chromatography using CH.sub.2Cl.sub.2, followed by 0.5:99.5
methanol/CH.sub.2Cl.sub.2, followed by a 1:99
methanol/CH.sub.2Cl.sub.2 solution as the solvent system to give
120 mg (42% yield) of the title compound as a white solid. TLC:
Rf=0.48 (3:97 methanol/CH.sub.2Cl.sub.2); HPLC: Rt=18.22 min,
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
[0683] D. Compound XXII (A=H, D'=Cyclopentylmethyl,
E=4-fluorophenyl, Hydrochloride Salt).
[0684] A solution of 266 mg of the resultant compound of Example
114C in ethyl acetate was treated at -20.degree. C. with HCl gas
for 20 min, during which time the temperature was allowed to warm
to 20.degree. C. Nitrogen was then bubbled through the mixture for
15 min and the solvent removed in vacuo to yield 224 mg of white
solid which was used directly for ensuing reaction.
[0685] E. Compound 114.
[0686] A solution of 31 mg of the resultant compound of Example
114D in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature, under an atmosphere of nitrogen, with 9 mg of allyl
chloroformate and 19 mg N,N-diisopropylethylamine. The mixture was
stirred for 3 h and then concentrated in vacuo. The residue was
taken up in ethyl acetate and washed in 0.5 N HCl and saturated
NaCl then dried over MgSO.sub.4, filtered, and concentrated in
vacuo to yield 34 mg of the title product as a white solid. TLC:
Rf=0.34 (5% diethyl ether in CH.sub.2Cl.sub.2). HPLC: Rt=17.21 min.
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 115
[0687] Compound 115.
[0688] A solution of 31 mg of the resultant compound of Example
114B in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 8 mg of ethyl
chloroformate and 19 mg N,N-diisopropylethylamine. The mixture was
stirred for 3 h and then concentrated in vacuo. The residue was
taken up in ethyl acetate and washed with 0.5 N HCl and saturated
NaCl then dried over MgSO.sub.4, filtered, and concentrated in
vacuo to yield 35 mg of the title product as a white solid. TLC:
Rf=0.32 (5% diethyl ether/CH.sub.2Cl.sub.2). HPLC: Rt=16.86 min.
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 116
[0689] A. Compound XXII (A=Tert-butoxycarbonyl,
D'=cyclopentylmethyl, E=4-chlorophenyl). The resultant compound of
Example 114B (252 mg) was reacted with 4-chlorobenzenesulfonyl
chloride (175 mg) in the manner described in Example 166A. Workup
and purification by silica gel chromatography using
EtOAc/CH.sub.2Cl.sub.2 as eluant yielded the product as a white
solid; (.sup.1H) NMR (CDCl.sub.3) consistent with structure.
[0690] B. Compound XXII (A=H, D'=Cyclopentylmethyl,
E=4-chlorophenyl, Hydrochloride Salt).
[0691] A solution of 320 mg of the resultant compound of Example
116A in 20 mL of EtOAc was treated with anhydrous HCl gas for 5
min. The reaction mixture was sparged with nitrogen then
concentrated in vacuo to yeld a white solid which was used directly
for subsequent reaction.
[0692] C. Compound 116.
[0693] To a solution of 63.4 mg of the resultant compound of
Example 116B in 1 mL of THF was added sequentially 54 .mu.L of
diisopropylethylamine and a solution of 39.9 mg of the resultant
compound of Example 48A in 1 mL THF. The mixture was stirred for 24
hours and then concentrated in vacuo. The residue was purified by
low pressure silica gel column chromatography using 20% EtOAc in
CH.sub.2Cl.sub.2 eluent to yield 0.62 g of the title compound. TLC:
Rf=0.71, 40% EtOAc/CH.sub.2Cl.sub.2, HPLC: Rt=16.88 min. (1H) NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 117
[0694] Compound 117.
[0695] A solution of 66.1 mg of the resultant compound of Example
116B in 1 mL of THF was treated sequentially with 56 .mu.L of
diisopropylethylamine and 19.3 .mu.L of allyl chloroformate. The
mixture was stirred for 4 hours and concentrated in vacuo. The
residue was taken into 50 mL of EtOAc and washed with 1.0 N HCl,
saturated sodium bicarbonate, brine, dried over magnesium sulfate,
filtered and concentrated. The residue was purified by low pressure
silica gel column chromatography using 20% EtOAc in hexane eluent
to yield 69.7 mg of the title compound. TLC: Rf=0.20, 20%
EtOAc/hexane. HPLC: Rt=17.83 min. (.sup.1H) NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 118
[0696] Compound 118.
[0697] To a solution of 65.3 mg of the resultant compound of
Example 116B in 1 mL of THF was added sequentially 55 .mu.L of
diisopropylethylamine and a solution of 49.2 mg of the resultant
compound of Example 82A in 1 mL THF. The mixture was stirred for 24
hours and concentrated in vacuo. The residue was purified by low
pressure silica gel column chromatography using 406 EtOAc in
CH.sub.2Cl.sub.2 as eluent followed by preparative reversed-phase
C.sub.18HPLC using a linear gradient of 40% to 80%
CH.sub.3CN/H.sub.2O for elution to yield 70.7 mg of the title
compound. TLC: Rf=0.27, 40% EtOAc/CH.sub.2Cl.sub.2, HPLC: Rt=14.85
min. (.sup.1H) NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 119
[0698] Compound 119.
[0699] A solution of 26 mg of the resultant compound of Example 81B
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 6 mg of ethyl
chloroformate and 15 mg N,N-diisopropylethylamine. The mixture was
stirred for 3 h and then concentrated in vacuo. The residue was
taken up in ethyl acetage and washed with 0.5 N HCl and saturated
NaCl then dried over MgSO.sub.4, filtered, and concentrated in
vacuo. The residue was purified by low pressure silica gel
chromatography using 5% diethyl ether/CH.sub.2Cl.sub.2 as eluent to
provide 26 mg of the title product as a white solid. TLC: Rf=0.19
(5% diethyl ether in CH.sub.2Cl.sub.2). HPLC: Rt=17.50 min.
.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 120
[0700] Compound 120.
[0701] A solution of 30 mg of the resultant compound of Example 40A
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 8 mg of ethyl
chloroformate and 18 mg N,N-diisopropylethylamine. The mixture was
stirred for 3 h and then concentrated in vacuo. The residue was
taken up in ethyl acetage and washed with 0.5 N HCl and saturated
NaCl then dried over MgSO.sub.4, filtered, and concentrated in
vacuo. The residue was purified by preparative thin layer silica
gel chromatography using ethyl acetate as eluent to yield 25 mg of
the title product as a white solid. TLC: Rf=0.60 (ethyl acetate).
HPLC: Rt=13.86 min. (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 121
[0702] Compound 121.
[0703] A solution of 26 mg of the resultant compound of Example 35A
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 7 mg of ethyl
chloroformate and 17 mg N,N-diisopropylethylamine. The mixture was
stirred for 3 h and then concentrated in vacuo. The residue was
taken up in ethyl acetate and washed with 0.5 N HCl and saturated
NaCl then dried over MgSO.sub.4, filtered, and concentrated in
vacuo. The residue was purified by low pressure silica gel
chromatography using 5% diethyl ether/CH.sub.2Cl.sub.2 as eluent to
provide 22 mg of the title product as a white solid. TLC: Rf=0.14
(5% diethyl ether/CH.sub.2Cl.sub.2). HPLC: Rt=15.95 min.
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 122
[0704] Compound 122.
[0705] A solution of 27 mg of the resultant compound of Example 35A
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 8 mg of allyl
chloroformate and 18 mg N,N-diisopropylethylamine. The mixture was
stirred for 3 h and then concentrated in vacuo. The residue was
taken up in ethyl acetate and washed with 0.5 N HCl and saturated
NaCl then dried over MgSO.sub.4, filtered, and concentrated in
vacuo. The residue was purified by low pressure silica gel
chromatography using 5% diethyl ether in CH.sub.2Cl.sub.2 as eluent
to provide 23 mg of the title product as a white solid. TLC:
Rf=0.33, 5% diethyl ether in CH.sub.2Cl.sub.2. HPLC: Rt=16.28 min.
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 123
[0706] A. Compound XXII (A=Tert-butoxycarbonyl, D'=Isobutyl,
E=3,4-dimethoxyphenyl).
[0707] To a solution of 401 mg (1.192 mmol) of the resultant
compound of Example 39A in CH.sub.2Cl.sub.2 (12 mL) was added
aqueous sodium bicarbonate (3 mL), solid sodium bicarbonate 130 mg
(1.549 mmol), and 3, 4-dimethoxybenzenesulfonyl chloride 33.8 mg,
(1.43 mmol). After 14 h, the resulting mixture was diluted with
EtOAc, washed with saturated brine, dried over magnesium sulfate,
filtered and concentrated in vacuo. The residue was purified by
flash chromatography using 5% to 25% EtOAc/CH.sub.2Cl.sub.2 eluent
to provide 440.1 mg of the title product. TLC: Rf=0.72, 20%
EtOAc/CH.sub.2Cl.sub.2.
[0708] B. Compound XXII (A=H, D'=Isobutyl, E=3,4-dimethoxyphenyl,
Hydrochloride Salt).
[0709] A solution of 440 mg (0.820 mmol) of the resultant compound
of Example 123A in EtOAc (15 mL) at -20.degree. C. was treated with
anhydrous HCl gas for 10 min. The ice bath was removed and after an
additional 15 min. the reaction mixture was sparged with nitrogen
then concentrated in vacuo to provide 610 mg of title product as
the HCl salt. TLC: Rf=0.44, 10% methanol/CH.sub.2Cl.sub.2.
[0710] C. Compound 123.
[0711] A solution of 38.9 mg of the resultant compound of Example
123B (0.170 mmol) in CH.sub.2Cl.sub.2 (3 mL) was treated
sequentially at ambient temperature under an atmosphere of nitrogen
with 0.049 mL (0.283 mmol) diisopropylethylamine and 66.9 mg (169.6
mmol) of the resultant compound of Example 48A. The mixture was
stirred for 16 h and then concentrated in vacuo. The residue was
taken up in CH.sub.2Cl.sub.2 and washed with saturated brine, dried
over magnesium sulfate, filtered, and concentrated in vacuo. The
residue was purified by low pressure silica gel column
chromatography using a gradient 10% to 25% diethyl ether in
CH.sub.2Cl.sub.2 cluent to yield 57.6 mg of the title product. TLC:
Rf=0.39, 25% diethyl ether/CH.sub.2Cl.sub.2. HPLC: Rt=14.3 min;
(.sup.1H) NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 124
[0712] A. Compound XXII (A=Tert-butoxycarbonyl, D'=Isobutyl, E=3,4
Difluorophenyl).
[0713] To a solution of 332.7 mg (0.989 mmol) of the resultant
compound of Example 39A in CH.sub.2Cl.sub.2 (12 mL) was added
aqueous sodium bicarbonate (3 mL), solid sodium bicarbonate 125 mg
(1.483 mmol), and 3,4 difluorobenzensulfonyl chloride 231 mg.
(1.088 mmol). After 14 h, the resulting mixture was diluted with
CH.sub.2Cl.sub.2, washed with saturated brine, dried over magnesium
sulfate, filtered and concentrated in vacuo. The residue was
purified by flash chromatography using 5% to 25% diethyl
ether/CH.sub.2Cl.sub.2 eluent to provide 313.6 mg of the title
product. (.sup.1H)-NMR (CDCl.sub.3) consistent with structure. B.
Compound XXII (A=H, D'=isobutyl, E=3,4 difluorophenyl,
hydrochloride salt). A solution of 312.6 mg (0.610 mmol) of the
resultant compound of Example 124A in EtOAc (15 mL) at -20.degree.
C. was treated with anhydrous HCl gas for 10 min. The ice bath was
removed and after an additional 15 min., the reaction mixture was
sparged with nitrogen then concentrated in vacuo to provide 280 mg
of title product as a white solid. TLC: Rf=0.46, 10%
methanol/CH.sub.2Cl.sub.2.
[0714] C. Compound 124.
[0715] To a solution of 64.7 mg of the resultant compound of
Example 124B (0.144 mmol) in CH.sub.2Cl.sub.2 (3 mL) was treated
sequentially at ambient temperature under an atmosphere of
nitrogen, with 0.050 mL (0.288 mmol) diisopropylethylamine and 39.6
mg (172.9 mmol)of the resultant compound of Example 48A. The
mixture was stirred for 16 h and then concentrated in vacuo. The
residue was taken up in CH.sub.2Cl.sub.2 and washed with saturated
brine, dried over magnesium sulfate, filtered, and concentrated in
vacuo. The residue was purified by low pressure silica gel column
chromatography using a gradient 5% to 20k diethyl ether in
CH.sub.2Cl.sub.2 eluent to yield 44 mg of the title product. TLC:
RF =0.54 25% diethyl ether/CH.sub.2Cl.sub.2, HPLC: Rt=15.4 min.
(.sup.1H) NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 125
[0716] Compound 125.
[0717] This compound was prepared from the resultant compound of
Example 146B in the manner described in Example 88. After workup
and purification by preparative reversed-phase C.sub.18HPLC using a
liner gradient of 35% to 100% CH.sub.3CN/H.sub.2O with 0.1% TFA as
eluant, 10.5 mg of the title compound was obtained as a white
solid. TLC: Rf=0.4, 4% MeOH/CH.sub.2Cl.sub.2, HPLC: Rt=14.06 min;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 126
[0718] A. Compound XXI (A=Tert-butoxycarbonyl, D'=Methyl,
A'=H).
[0719] To a solution compound XX (1.7 mmol) in ethanol (20 mL) was
added methylamine gas, at ambient temperature, for 30 min. The
solution was stirred overnight, then concentrated under reduced
pressure to give 0.47 g of the title compound which was used
without subsequent purification. TLC: Rf=0.19, 1:10:90
NH.sub.4OH/methanol/CH.sub.2Cl.sub.2, .sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
[0720] B. Compound 126.
[0721] To a solution of the product of Example 126A (0.15 g, 0.51
mmol) in CH.sub.2Cl.sub.2 (10 mL) was added a saturated solution of
sodium bicarbonate (3 mL), followed by addition of solid sodium
bicarbonate (90 mg, 1.1 mmol), followed by addition of
3,4-dichlorobenzenesulfonyl chloride (0.25 g, 1.0 mmol). The
mixture was stirred at ambient temperature overnight. The organics
were extracted into 100 mL CH.sub.2Cl.sub.2, dried over anhydrous,
MgSO.sub.4, concentrated under reduced pressure then purified via
medium pressure silica gel chromatography using a gradient system
of CH.sub.2Cl.sub.2 followed by 5:95 ether/CH.sub.2Cl.sub.2. The
title compound was obtained as a colorless foam 210 mg. TLC:
Rf=0.42 (3:97 methanol/CH.sub.2Cl.sub.2), HPLC: Rt=17.2 min;
.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 127
[0722] Compound 127.
[0723] To a solution of the product of Example 126A (0.15 g, 0.51
mmol) in CH.sub.2Cl.sub.2 (10 mL) was added a saturated solution of
sodium bicarbonate (3 mL), followed by addition of solid sodium
bicarbonate (100 mg, 1.0 mmol), followed by addition of
4-fluorobenzenesulfonyl chloride (0.20 g, 1.0 mmol). The mixture
was stirred at ambient temperature overnight. The organics were
extracted into 100 mL CH.sub.2Cl.sub.2, dried over anhydrous,
MgSO.sub.4, concentrated under reduced pressure then purified via
medium pressure silica gel chromatography using a gradient system
of CH.sub.2Cl.sub.2 followed by 5:95 ether/CH.sub.2Cl.sub.2. The
title compound was obtained as a white solid 104 mg. TLC: Rf=0.36,
3:97 methanol/CH.sub.2Cl.sub.2, HPLC: Rt=15.86 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 128
[0724] Compound 128.
[0725] To a solution of the product of Example 126A (0.15 g, 0.51
mmol) in CH.sub.2Cl.sub.2 (6 mL) was added a saturated solution of
sodium bicarbonate (3 mL), followed by addition of solid sodium
bicarbonate (90 mg, 1.0 mmol), followed by addition of
acetamidobenzenesulfonyl chloride (0.24 g, 1.02 mmol). The mixture
was stirred at ambient temperature overnight. The organics were
extracted into 100 mL CH.sub.2Cl.sub.2, dried over anhydrous,
MgSO.sub.4, concentrated under reduced pressure then purified via
medium pressure silica gel chromatography using a gradient system
of CH.sub.2Cl.sub.2 followed by 5:95 EtOAc/CH.sub.2Cl.sub.2,
followed by 10:90 EtOAc/CH.sub.2Cl.sub.2. The title compound was
obtained as 244 mg of white solid. TLC: Rf=0.13, 3:97
methanol/CH.sub.2Cl.sub.2, HPLC: Rt=13.47 min; (1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 129
[0726] A. Compound XXI (A=Tert-butoxycarbonyl,
D'=(2-tetrahydrofuryl)-meth- yl, A'=H).
[0727] To a solution compound XX (3.3 mmol) in ethanol (30 mL) was
added tetrahydrofurfurylamine (1.03 mL, 10 mmol). The mixture was
warmed to 85.degree. C. and stirred overnight. The solution was
filtered and the solution concentrated under reduced pressure to
give 1.29 g of the title compound which was used without subsequent
purification. TLC: Rf=0.52, 1:10:90
NH.sub.4OH/methanol/CH.sub.2Cl.sub.2.
[0728] B. Compound 129.
[0729] To a solution of the resultant compound of Example 129A (200
mg, 0.55 mmol) in CH.sub.2Cl.sub.2 (6 mL) was added
4-fluorobenzenesulfonyl chloride (320 mg, 1.6 mmol) followed a
saturated solution of sodium bicarbonate (3 mL) and solid sodium
bicarbonate (0.1 g, 1.2 mmol). The mixture was stirred at ambient
temperature overnight. The solution was diluted with 100 mL
CH.sub.2C.sub.2, the organics separated, dried over anhydrous
MgSO.sub.4, and the organics concentrated under reduced pressure.
The crude product was purified via medium pressure liquid
chromatography using a gradient solvent system of CH.sub.2Cl.sub.2
followed by 5:95 ether/CH.sub.2Cl.sub.2 followed by a 10:90
ether/CH.sub.2Cl.sub.2 solution to give 130 mg of the title
compound as a white solid. TLC: Rf=0.35, 3:97
methanol/CH.sub.2Cl.sub.2, HPLC: Rt=16.37 min, (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 130
[0730] A. Compound XXI (A=Tert-butoxycarbonyl, D'=(Isobutenyl,
A'=H)).
[0731] To a solution compound XX (A=tert-butoxycarbonyl) (2.5 mmol)
in ethanol (30 mL) was added a solution 2-methylallylamine
hydrochloride (1.34 g, 12.5 mmol) and KOH (0.70 g, 12.5 mmol) in
ethanol (20 mL). The mixture stirred 30 min at ambient temperature.
The solutions were combined and heated to 85.degree. C. for 24 h.
The solution was filtered and concentrated under reduced pressure
to give 0.82 g of the title compound which was used without
subsequent purification. TLC: Rf=0.45, 1:10:90 concentrated
NH.sub.4OH/methanol/CH.sub.2Cl.sub.2.
[0732] B. Compound 130.
[0733] To a solution of the product of Example 130A (0.20 g, 0.60
mmol) in CH.sub.2Cl.sub.2 (6 mL) was added a saturated solution of
sodium bicarbonate (3 mL), followed by solid sodium bicarbonate
(0.1 g, 1.2 mmol) and then p-fluorobenzenesulfonyl chloride (0.35
g, 1.78 mmol). The mixture was stirred at ambient temperature for
24 h. The organics were extracted into 100 mL CH.sub.2Cl.sub.2,
dried over anhydrous MgSO.sub.4, concentrated under reduced
pressure then purified via medium pressure silica gel
chromatography using a gradient system of CH.sub.2Cl.sub.2,
followed by 1:99 methanol/CH.sub.2Cl.sub.2. The title compound was
obtained as a white solid 180 mg. TLC: Rf=0.35, 3:97
methanol/CH.sub.2Cl.sub.2, HPLC: Rt=16.82 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 131
[0734] Compound 131.
[0735] To a solution of the resultant compound of Example 130A (200
mg, 0.60 mmol) in CH.sub.2Cl.sub.2 (6 mL) was added
4-acetamidobenzenesulfony- l chloride (410 mg, 1.76 mmol), followed
by a saturated solution of sodium bicarbonate (3 mL) and solid
sodium bicarbonate (0.1 g, 1.2 mmol). The mixture was stirred at
ambient temperature overnight. The solution was diluted with 100 mL
CH.sub.2Cl.sub.2, the organics separated, dried over anhydrous
MgSO.sub.4, and the organics concentrated under reduced pressure.
The crude product was purified via medium pressure liquid
chromatography using a gradient solvent system of CH.sub.2Cl.sub.2,
followed by 30:70 EtOAc/CH.sub.2Cl.sub.2 solution to give 140 mg of
the title compound as a white solid. TLC: Rf=0.19, 3:97
methanol/CH.sub.2Cl.sub.2HPLC: Rt=15.06 min, (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 132
[0736] A. Compound XXII (A=H, D'=(2-tetrahydrofuryl)-methyl,
E=4-fluorophenyl, hydrochloride salt). To a solution of the
resultant compound of Example 129B (30 mg, 0.057 mmol) in EtOAc (3
mL) was added 30% w/w HCl in EtOAc (1 mL). The mixture was stirred
overnight at ambient temperature. The solution was concentrated
under reduced pressure to give 16 mg of the title compound as a
white solid which was used without subsequent purification. TLC:
Rf=0.60 (1:10:90 NH.sub.4OH/methanol/CH.sub- .2Cl.sub.2).
[0737] B. Compound 132.
[0738] To a solution of the resultant compound of Example 132A (16
mg) in CH.sub.2Cl.sub.2 (5 mL) was added triethylamine (0.1 mL,
0.72 mmol) followed by the compound of Example 48A (20 mg, 0.09
mmol). The mixture was stirred at ambient temperature for 24 hours.
The solution was concentrated under reduced pressure and the crude
product purified via medium pressure colum chromatography using
20:80 EtOAc/CH.sub.2C12 as the solvent system to give 7.4 mg.
Rf=0.37 (3:97 methanol/CH.sub.2Cl.sub.2), HPLC: Rt=14.19 min,
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 133
[0739] A. Compound XXII (A=Tert-butoxycarbonyl,
D'=(2-tetrahydrofuryl)-met- hyl, E=4-acetamidophenyl).
[0740] To a solution of the resultant compound of Example 129A (200
mg, 0.55 mmol) in CH.sub.2Cl.sub.2 (6 mL) was added
4-acetamidobenzenesulfony- l chloride (380 mg, 1.6 mmol) followed
by a saturated solution of sodium bicarbonate (3 mL) and solid
sodium bicarbonate (0.1 g, 1.2 mmol). the mixture was stirred at
ambient temperature overnight. The solution was diluted with 100 mL
CH.sub.2Cl.sub.2, the organics separated, dried over anhydrous.
MgSO.sub.41 and the organics concentrated under reduced pressure.
The crude product was purified via medium pressure liquid
chromatography using a gradient solvent system of CH.sub.2Cl.sub.2,
followed by 10:90 EtOAc/CH.sub.2Cl.sub.2, followed by a 30:70
EtOAc/CH.sub.2Cl.sub.2 solution to give 120 mg of the title
compound as a white solid. TLC: Rf=0.13, 3:97
methanol/CH.sub.2Cl.sub.2, (.sup.1H)-NMR (CDCl.sub.3) consistent
with structure.
[0741] B. Compound XXII (A=H, D'=(2-tetrahydrofuryl)-methyl,
E=4-acetamidophenyl, Hydrochloride Salt).
[0742] To a solution of the resultant compound of Example 133A (120
mg 0.22 mmol) in EtOAc (5 mL) was added 30% w/w 5HCl in EtOAc (2
mL). The mixture was stirred overnight at ambient temperature. The
solution was concentrated under reduced pressure to give the title
compound which was used without subsequent purification. TLC:
Rf=0.50, 1:10:90 NH.sub.4OH/methanol/CH.sub.2Cl.sub.2.
[0743] C. Compound 133.
[0744] To a solution of the resultant compound of Example 133B in
CH.sub.2Cl.sub.2 (S mL) was added triethylamine (0.2 mL, 1.4 mmol)
followed by the compound of Example 48A (73 mg, 0.32 mmol). The
mixture was stirred at ambient temperature for 24 hours. The
solution was concentrated under reduced pressure and the crude
product purified via medium pressure column chromatography using a
gradient solvent system of CH.sub.2Cl.sub.2, followed by 1:99
methanol/CH.sub.2Cl.sub.2, followed by 3:97
methanol/CH.sub.2Cl.sub.2 as the solvent system to give 87.8 mg.
Rf=0.09, 3:97 methanol/CH.sub.2Cl.sub.2, HPLC:
[0745] Rt=12.53 min, (.sub.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 134
[0746] A. Compound XXII (A=H, D'=Isobutenyl, E=4-acetamidophenyl,
Hydrochloride Salt).
[0747] To a solution of the resultant compound of Example 131 (40
mg 0.075 mmol) in EtOAc (5 mL) was added 306 w/w HCl in EtOAc (2
mL). The mixture was stirred overnight at ambient temperature. The
solution was concentrated under reduced pressure to give the title
compound, which was used without subsequent purification. TLC:
Rf=0.38, 1:10:90 NH.sub.4OH/methanol/CH.sub.2Cl.sub.2.
[0748] B. Compound 134.
[0749] To a solution of the resultant compound of Example 134A in
CH.sub.2Cl.sub.2 (5 mL) was added triethylamine (0.1 mL, 0.72
mmol), followed by the compound of Example 48A (26 mg, 0.11 mmol).
The mixture was stirred at ambient temperature for 24 hours. The
solution was concentrated under reduced pressure and the crude
product purified via medium pressure column chromatography using a
gradient solvent system of CH.sub.2Cl.sub.2, followed by 1:99
methanol/CH.sub.2Cl.sub.2, followed by 3:97
methanol/CH.sub.2Cl.sub.2 as the solvent system to give 10.1 mg of
the title compound. Rf=0.11 (3:97 methanol/CH.sub.2Cl.sub.2), HPLC:
Rt=12.86 min, (.sub.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 135
[0750] A. Compound XXI (A=H, D'=(Isobutenyl, E=4-fluoropheynl,
Hydrochloride Salt).
[0751] To a solution of the resultant compound of Example 130B (50
mg, 0.10 mmol) in EtOAc (5 mL) was added 30% w/w HCl in EtOAc (1
mL). The mixture was stirred overnight at ambient temperature. The
solution was concentrated under reduced pressure to give the title
compound which was used without subsequent purification. TLC:
Rf=0.48, 1:10:90 NH.sub.4OH/methanol/CH.sub.2Cl.sub.2.
[0752] B. Compound 135.
[0753] To a solution of the resultant compound of Example 135A in
CH.sub.2Cl.sub.2 (5 mL) was added triethylamine (0.1 mL, 0.72
mmol), followed by the compound of Example 48A (35 mg, 0.15 mmol).
The mixture was stirred at ambient temperature for 24 hours. The
solution was concentrated under reduced pressure and the crude
product purified via medium pressure column chromatography using a
gradient solvent system of CH.sub.2Cl.sub.2 followed by 20:80
EtOAc/CH.sub.2Cl.sub.2 as the solvent system to give 12 mg.
Rf=0.34, 3:97 methanol/CH.sub.2Cl.sub.2, HPLC: Rt=14.64 min,
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 136
[0754] A. Compound XXI (A=Tert-butoxycarbonyl, D'=2-furfuryl,
A'=H). To a solution compound XX (2.5 mmol) in ethanol (30 mL) was
added furfurylamine (0.67 mL, 7.5 mmol) and the mixture was heated
to 85.degree. C. for 24 h. The solution was filtered and
concentrated under reduced pressure to give 0.80 g of the title
compound which was used without subsequent purification. TLC:
Rf=0.38, 1:10:90 concentrated
NH.sub.4OH/methanol/CH.sub.2Cl.sub.2.
[0755] B. Compound XXII (A=Tert-butoxycarbonyl, D'=2-furyl,
E=4-fluorophenyl).
[0756] To a solution of the product of Example 136A (0.20 g, 0.60
mmol) in CH.sub.2Cl.sub.2 (6 mL) was added a saturated solution of
sodium bicarbonate (3 mL), followed by addition of solid sodium
bicarbonate (0.1 g, 1.2 mmol), then p-fluorobenzenesulfonyl
chloride (0.32 g, 1.6 mmol). The mixture was stirred at ambient
temperature for 24 h. The organics were extracted into 100 mL
CH.sub.2Cl.sub.2, dried over anhydrous MgSO.sub.4, concentrated
under reduced pressure, then purified via medium pressure silica
gel chromatography using a gradient system of CH.sub.2Cl.sub.2,
followed by 1:99 methanol/CH.sub.2Cl.sub.2. The title compound was
obtained as a white solid (86.1 mg). TLC: Rf=0.17, 3:97
methanol/CH.sub.2Cl.sub.2, HPLC: Rt=16.5 min; (.sub.1H)-NMR
(CDCl.sub.3) consistent with structure.
[0757] C. Compound XXII (A=H, D'=2-furyl, E=4-fluorophenyl,
Hydrochloride Salt).
[0758] To a solution of the resultant compound of Example 136B (16
mg, 0.031 mmol) in EtOAc (3 mL) was added 30% w/w HCl in EtOAc (1
mL). The mixture was stirred overnight at ambient temperature. The
solution was concentrated under reduced pressure to give the title
compound, which was used without subsequent purification. TLC:
Rf=0.48, 1:10:90 NH.sub.4OH/methanol/CH.sub.2Cl.sub.2.
[0759] D. Compound 136.
[0760] To a solution of the resultant compound of Example 136C in
CH.sub.2Cl.sub.2 (5 mL) was added triethylamine (0.1 mL, 0.72
mmol), followed by the resultant compound of Example 48A (11 mg,
0.05 mmol). The mixture was stirred at ambient temperature for 24
hours. The solution was concentrated under reduced pressure and the
crude product purified via medium pressure column chromatography
using a gradient solvent system of CH.sub.2Cl.sub.2 followed by
20:80 EtOAc/CH.sub.2Cl.sub.2 as the solvent system to give 4.9 mg.
TLC: Rf=0.28, (3:97 methanol/CH.sub.2Cl.sub.2, HPLC: Rt=14.57 min,
(.sub.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 137
[0761] A. Compound XXII (A=Tert-butoxycarbonyl, D'=2-furyl,
E=4-acetamidophenyl).
[0762] To a solution of the resultant compound of Example 136B (200
mg, 0.55 mmol) in CH.sub.2Cl.sub.2 (6 mL) was added
4-acetamidobenzenesulfony- l chloride (390 mg, 1.7 mmol) followed
by saturated solution of sodium bicarbonate (3 mL) and solid sodium
bicarbonate (0.1 g, 1.2 mmol). The mixture was stirred at ambient
temperature overnight. The solution was diluted with 100 mL
CH.sub.2Cl.sub.2, the organics separated, dried over anhydrous.
MgSO.sub.41 and the organics concentrated under reduced pressure.
The crude product was purified via medium pressure liquid
chromatography using a gradient solvent system of CH.sub.2Cl.sub.2,
followed by 10:90 EtOAc/CH.sub.2Cl.sub.2, followed by a 30:70
EtOAc/CH.sub.2Cl.sub.2 solution to give 100 mg of the title
compound as a white solid. TLC: Rf=0.19, 3:97
methanol/CH.sub.2Cl.sub.2, (.sup.1H)-NMR (CDCl.sub.3) consistent
with structure.
[0763] B. Compound XXII (A=H, D'=2-furyl, E=4-acetamidophenyl,
Hydrochloride Salt).
[0764] To a solution of the resultant compount of Example 137A (30
mg, 0.054 mmol) in EtOAc (3 mL) was added 30% w/w HCl in EtOAc (1
mL). The mixture was stirred overnight at ambient temperature. The
solution was concentrated under reduced pressure to give the title
compound which was used without subsequent purification. TLC:
Rf=0.37 (1:10:90 NH.sub.4OH/methanol/CH.sub.2Cl.sub.2).
[0765] C. Compound 137.
[0766] To a solution of the resultant compound of Example 137a in
CH.sub.2Cl.sub.2 (5 mL) was added triethylamine (0.1 mL, 0.72 mmol)
followed by the compound of Example 48A (19 mg, 0.083 mmol). The
mixture was stirred at ambient temperature for 24 hours. The
solution was concentrated under reduced pressure and the crude
product purified via medium pressure column chromatography using a
gradient solvent system of CH.sub.2Cl.sub.2, followed by 1:99
methanol/CH.sub.2Cl.sub.2, followed by 3:97
methanol/CH.sub.2Cl.sub.2 as the solvent system to give 8.5 mg of
the title compound. TLC: Rf=0.11 (3:97 methanol/CH.sub.2Cl.sub.2),
HPLC: Rt=12.69 min; (.sub.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 138
[0767] Compound 138.
[0768] A solution of 75 mg of the resultant compound of Example 51D
and 45 mg of 3-chlorobenzenesulfonyl chloride were reacted in the
manner described in Example 60. After workup and purification by
preparative reversed-phase C.sub.18HPLC using a linear gradient of
35% to 100% CH.sub.3CN/H.sub.2O with 0.1% TFA as eluant, 29.7 mg of
the title compound was obtained. TLC: Rf=0.3, 4%
MeOH/CH.sub.2Cl.sub.2, HPLC: Rt=15.83 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 139
[0769] Compound 139.
[0770] To a solution of 67.9 mg of the resultant compount of
Example 116B in 1 mL of THF was added sequentially, 57 .mu.L of
diisopropylethylamine and a solution of 52.6 mg of the resultant
compound of Example 109C in 1 mL THF. The mixture was stirred for
24 hours and concentrated in vacuo. The residue was purified by
preparative thick layer silica gel chromatography using 7% methanol
in CH.sub.2Cl.sub.2 eluent to yield 70.0 mg of the title compound.
TLC: Rf=0.30, 5% methanol/CH.sub.2Cl.sub.2, HPLC: Rt=15.78 min;
(.sup.1H) NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 140
[0771] A. 3(S)-amino-2(syn)-hydroxy-4-phenyl-1-chlorobutane Formate
Salt.
[0772] To a slurry of 16.33 g of 10% palladium on carbon (25% by
weight) in methanol and tetrahydrofuran (400 mL, 1:1) was added,
under N.sub.2, 65.35 g of
3(S)-N-(-benzyloxycarbonyl)-amino-1-chloro-2(syn)-hydroxy-4-ph-
enylbutane (195.77 mmol) as a solution in methanol and
tetrahydrofuran (1.2 L). To this slurry was added 540 mL of formic
acid. After 15 h, the reaction mixture was filtered through
diatomaceous earth and concentrated to dryness. The resultant oil
was slurried in toluene and evaporated, then triturated
sequentially with diethyl ether and CH.sub.2Cl.sub.2 to provide
47.64 g of product as a granular tan solid. TLC: Rf=0.17, 5% acetic
acid/ethyl acetate.
[0773] B.
3(S)-N-(3(S)-tetrahydrofuryloxycarbonyl)-amino-1-chloro-2(syn)-h-
ydroxy-4-phenylbutane.
[0774] To a solution of the resultant compound of Example 140A
(1.97 g, 7.95 mmol) in CH.sub.2Cl.sub.2 (20 mL) was added a
saturated solution of sodium bicarbonate (5 mL), followed by solid
sodium bicarbonate (1.33 g, 17.9 mmol), and the resultant compound
of Example 48A (2.0 g, 8.7 mmol). The mixture was stirred at
ambient temperature overnight. The solution was diluted with 200 mL
CH.sub.2Cl.sub.2, the organics separated, dried over anhydrous
MgSO.sub.4, and concentrated under reduced pressure. The residue
was recrystallized from ethyl acetate/hexane to give 1.01 g of the
title compound as a white solid. TLC: Rf=0.35, 3:97
methanol/CH.sub.2Cl.sub.2. (.sup.1H)-NMR (CDCl.sub.3) consistent
with structure.
[0775] C. Compound XX (A=3(s)-tetrahydrofuryloxycarbonyl).
[0776] To a solution of the resultant compound of Example 140B (1.0
g, 3.2 mmol) in absolute ethanol (15 mL) was added solid KOH (0.21
g, 3.8 mmol). The mixture was stirred at ambient temperature for
1.0 h. The solution was filtered through a pad of Celite then
concentrated under reduced pressure. The residue was taken up in
ether (100 mL), washed with brine, dried over MgSO.sub.4, the
concentrated under reduced pressure to give 0.88 g of the title
compound as a white solid. TLC: Rf=0.49 (3:97
methanol/CH.sub.2Cl.sub.2), (.sup.1H)-NMR (CDCl.sub.3) consistent
with structure.
[0777] D. Compound XXI (A=(S)-3-tetrahydrofuryloxycarbonyl,
D'=Cyclopentylmethyl, A'=H).
[0778] The resultant compound of Example 140C (0.88 g, 3.2 mmol)
was added to the resultant compound of Example 114A (5.0 g, 50.4
mmol) and stirred for 24 h at ambient temperature. The solution was
concentrated by distillation under reduced pressure. The residue
was triturated with hexane and the solid collected by suction
filtration and washed with hexane to give 0.93 g of the title
compound. TLC: Rf=0.44, 1:10:90 concentrated
NH.sub.4OH/methanol/CH.sub.2Cl.sub.2; (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
[0779] E. Compound 140.
[0780] To a solution of the resultant compound of Example 140D
(0.93 g, 2.47 mmol) in CH.sub.2Cl.sub.2 (20 mL) was added a
saturated solution of sodium bicarbonate (5 mL) followed by
addition of solid sodium bicarbonate (0.42 g, 4.94 mmol) and
4-methoxybenzenesulfonyl chloride (0.61 g, 2.96 mmol). the mixture
was stirred at ambient temperature for 4 hours. The solution was
diluted with 200 mL CH.sub.2Cl.sub.2, the organics separated, dried
over anhydrous MgSO.sub.4, and the organics concentrated under
reduced pressure. The crude product was purified via medium
pressure liquid chromatography using CH.sub.2Cl.sub.2 followed by
1:99 methanol/CH.sub.2Cl.sub.2 solution as the eluent system to
give 1.28 g of the title compound as a white solid. TLC: Rf=0.26,
3:97 methanol/CH.sub.2Cl.sub.2, HPLC: Rt=15.66 min, H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 141
[0781] A. Compound XXII (A=H, D'=Cyclopentylmethyl,
E=4-methoxyphenyl, Hydrochloride Salt).
[0782] A solution of 71.3 mg of the resultant compound of Example
166A in EtOAc (25 mL) at 0.degree. C. was treated with anhydrous
HCl gas for 10 min., and allowed to stand for 12 h while warming to
ambient temperature, then concentrated under reduced pressure and
the resulting white solid used without purification for subsequent
reaction.
[0783] B. Compound 141.
[0784] The resultant compound of Example 141A (0.134 mmol) was
reacted with allyl chloroformate in the manner described in Example
87B. After concentration of the mixture in vacuo and workup, the
residue was purified by thick layer silica gel chromatography using
5% MeOH/CH.sub.2Cl.sub.2 as eluant followed by preparative
reversed-phase C.sub.18HPLC using a linear gradient of 35% to 100%
CH.sub.3CN/H.sub.2O with 0.1% TFA as eluant to obtain 21.6 mg of
the title compound. TLC: Rf=0.45, 5% MeOH/CH.sub.2Cl.sub.2, HPLC:
Rt=16.96 min.
EXAMPLE 142
[0785] Compound 142.
[0786] To a solution of 4.0 g of the resultant compound of Example
141A in 45 mL of THF was added sequentially, 1.96 mL of
diisopropylethylamine and a solution of 2.68 g of the resultant
compound of Example 82A in 45 mL THF. The mixture was stirred for
24 hours and concentrated in vacuo. The residue was taken up in
CH.sub.2Cl.sub.2, washed with saturated sodium bicarbonate and
brine, dried over magnesium sulfate, filtered and concentrated in
vacuo. The residue was purified by low pressure silica gel column
chromatography using 20% to 40% EtOAc in hexane eluent to yield
3.69 g of the title compound. TLC: Rf=0.41, 50%
EtOAc/CH.sub.2Cl.sub.2.
EXAMPLE 143
[0787] Compound 143.
[0788] A solution of 3.69 g of the resultant compound of Example
142 in 100 mL of ethyl ether was treated with anhydrous HCl gas for
10 min. The reaction mixture was sparged with nitrogen then
filtered. The solid was taken up in methanol and concentrated to
yield 3.71 g of the title compound. TLC: Rf=0.62, 90/10/1
CH.sub.2Cl.sub.2/MeOH/AcOH, HPLC: Rt=13.87 min. (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 145
[0789] A. Compound XXII (A=Tert-butoxycarbonyl, D'=Isobutyl,
E=2-(5-isoxazoy-3-yl)-thiophene).
[0790] To a solution of 342.5 mg (1.02 mmol) of the resultant
compound of Example 39A in CH.sub.2Cl.sub.2 (8 mL) was added
aqueous sodium bicarbonate (2 mL), solid sodium bicarbonate 257 mg
(3.1 mmol), and 5-(isoxazol-3-yl)-thiophenesulfonyl chloride 254.2
mg, (1.02 mmol). After 14 h, the resulting mixture was diluted with
CH.sub.2Cl.sub.2, washed with saturated brine, dried over magnesium
sulfate, filtered and concentrated in vacuo. The residue was
purified by flash chromatography using 5% to 25%
EtOAc/CH.sub.2Cl.sub.2 eluent and recrystallized from ether
CH.sub.2Cl.sub.2 to provide 228.6 mg of the title product.
.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
[0791] B. Compound XXII (A-H, D'=Isobutyl,
E=2-(5-isoxazoy-3-ly)-thiopene, Hydrochloride Salt).
[0792] A solution of 228.6 mg (0.416 mmol) of the resultant
compound of Example 145A in EtOAc (15 mL) at -20.degree. C. was
treated with anhydrous HCl gas for 10 min. The ice bath was removed
and after an additional 15 min, the reaction mixture was sparged
with nitrogen then concentrated in vacuo to provide 223.6 mg of
title product as the HCl salt. TLC: Rf=0.48, 10%
methanol/CH.sub.2Cl.sub.2.
[0793] C. Compound 145.
[0794] A solution of 78.5 mg of the resultant compound of Example
145B (0.162 mmol) in CH.sub.2Cl.sub.2 (3 mL) was treated
sequentially at ambient temperature under an atmosphere of nitrogen
with 0.07 mL (0.408 mmol) diisopropylethylamine and 55.6 mg (0.243
mmol) of the resultant compound of Example 48A. The mixture was
stirred 16 h and then concentrated in vacuo. The residue was taken
up in CH.sub.2Cl.sub.2 and washed with saturated brine, dried over
magnesium sulfate, filtered, and concentrated in vacuo. The residue
was purified by preparative HPLC to yield 48.7 mg of the title
product. TLC: Rf=0.36, 25% EtAOc/CH.sub.2Cl.sub.2, HPLC: Rt=15.2
min; (.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 146
[0795] A. Compound XXI (A=Tert-butoxycarbonyl,
D'=Cyclopentylmethyl, E=4-acetamidophenyl).
[0796] To a solution of the resultant compound of Example 114B (300
mg, 0.83 mmol) in CH.sub.2Cl.sub.2 (15 mL) was added
4-acetamidobenzenesulfon- yl chloride (580 mg, 2.48 mmol) followed
by the addition of a saturated solution of sodium bicarbonate (4
mL) and solid sodium bicarbonate (0.14 g, 1.67 mmol). The mixture
was stirred at ambient temperature overnight. The solution was
diluted with 150 mL CH.sub.2Cl.sub.2, the organics separated, dried
over anhydrous. MgSO.sub.4 and the organics concentrated under
reduced pressure. The crude product was purified via medium
pressure liquid chromatography using a gradient solvent system of
CH.sub.2Cl.sub.2, followed by 5:95 EtOAc/CH.sub.2Cl.sub.2, followed
by 10:90 EtOAc/CH.sub.2Cl.sub.2 solution to give 310 mg of the
title compound as a white solid. TLC: Rf=0.10, 3:97
methanol/CH.sub.2Cl.sub.2, HPLC: Rt=15.96 min, (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
[0797] B. Compound XXII (A=H, D'=Cyclopentylmethyl,
E=4-acetamidophenyl, Hydrochloride Salt).
[0798] To a solution of the resultant compound of Example 146A (210
mg, 0.38 mmol) was added 30% w/w HCl in EtOAc (15 mL). The mixture
was stirred for 1 hour at ambient temperature. The solution was
concentrated under reduced pressure to give 180 mg of the title
compound which was used without subsequent purification. TLC:
Rf=0.14, 1:10:90 NH.sub.4OH/methanol/CH.sub.2Cl.sub.2.
[0799] C. Compound XXII (A=allyloxycarbonyl, D'=Cyclopentylmethyl,
E=4-acetamidophenyl).
[0800] To a solution of the resultant compound of Example 146B (100
mg, 0.20 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added triethylamine
(0.1 mL, 0.72 mmol), followed by allylchloroformate (0.04 mL, 0.3
mmol). The mixture was stirred at ambient temperature for 24 hours.
The solution was diluted with 150 mL CH.sub.2Cl.sub.2, washed with
water, dried over anhydrous MgSO.sub.4, and the organics
concentrated under reduced pressure. The crude product was purified
via medium pressure column chromatography using a gradient solvent
system of CH.sub.2Cl.sub.2, followed by 1:99
methanol/CH.sub.2Cl.sub.2, followed by 3:97
methanol/CH.sub.2Cl.sub.2 as the solvent system to give 103 mg.
Rf=0.22, 3:97 methanol/CH.sub.2Cl.sub.2HPLC: Rt=15.29 min,
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 147
[0801] Compound 147.
[0802] To a solution of the resultant compound of Example 146B (80
mg, 0.16 mmol) in CH.sub.2Cl.sub.2(5 mL) was added triethylamine
(0.07 mL, 0.48 mmol), followed by slow addition over 3 hours of the
resultant compound of Example 82A (53 mg, 0.19 mmol) as a solution
in CH.sub.2Cl.sub.2 (3 mL). The mixture was stirred at ambient
temperature for 24 hours. The solution was diluted with 100 mL
CH.sub.2Cl.sub.2 washed with water, dried over anhydrous
MgSO.sub.4, and the organics concentrated under reduced pressure.
The crude product was purified via medium pressure column
chromatography using a gradient solvent system of CH.sub.2Cl.sub.2,
followed by 1:99 methanol/CH.sub.2Cl.sub.2, followed by 2:98
methanol/CH.sub.2Cl.sub.2 as the solvent system to give 71.7 mg of
the title compound. Rf=0.06, 3:97 methanol/CH.sub.2Cl.sub.2, HPLC:
Rt=12.61 min, (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 148
[0803] A. Compound XXII (A=Tert-butoxycarbonyl,
D'=Cyclopentylmethyl, E=Phenyl).
[0804] A solution of 297 mg of the resultant compound of Example
114B in 4:1 CH.sub.2Cl.sub.2/saturated aqueous NaHCO.sub.3 was
treated sequentially, at ambient temperature under an atmosphere of
nitrogen, with 217 mg of benzenesulfonyl chloride and 103 mg of
sodium bicarbonate. The mixture was stirred for 6 h, diluted with
CH.sub.2Cl.sub.2, washed with saturated NaCl then dried over
MgSO.sub.4, filtered, and concentrated in vacuo to yield 426 mg of
the title product as a white solid. TLC: Rf=0.32, 5% diethyl
ether/CH.sub.2Cl.sub.2. (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
[0805] B. Compound XXII (A=H, D'=Cyclopentylmethyl, E=Phenyl,
Hydrochloride Salt).
[0806] A solution of 400 mg of the resultant compound of Example
148A in ethyl acetate was treated at -20.degree. C. with HCl gas
for 20 min, during which time the temperature was allowed to warm
to 20.degree. C. Nitrogen was then bubbled through the mixture for
15 min and solvent removed in vacuo to yield 349 mg of white solid
which was used directly for the ensuing reaction.
[0807] C. Compound 148.
[0808] A solution of 40 mg of the resultant compound of Example
148B in CH.sub.2Cl.sub.2 was added, at ambient temperature under an
atmosphere of nitrogen, to a solution of 31 mg of the resultant
compound of Example 48A and 35 mg N,N-diisopropylethylamine in
CH.sub.2Cl.sub.2. The mixture was stirred for 14 h, diluted with
CH.sub.2Cl.sub.2, washed with saturated NaHCO.sub.3 and saturated
NaCl, then dried over MgSO.sub.4, filtered, and concentrated in
vacuo. The residue was purified by low pressure silica gel
chromatography using 20% diethyl ether/CH.sub.2Cl.sub.2 as eluent
to provide 45 mg of the title product as a white solid. TLC:
Rf=0.46, 20% diethyl ether/CH.sub.2Cl.sub.2. HPLC: Rt=15.78 min.
.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 149
[0809] A. Compound XXII (A=Tert-butoxycarbonyl,
D'=Cyclopentymethyl, E=3-pyridyl).
[0810] To a solution of 153 mg (0.422 mmol) of the resultant
compound of Example 114B in CH.sub.2Cl.sub.2 (4 mL) was added
aqueous sodium bicarbonate (1 mL), solid sodium bicarbonate 141.7
mg (1.69 mmol), and the resultant compound of Example 144A 156.1
mg. (0.879 mmol). After 14 h, the resulting mixture was diluted
with CH.sub.2Cl.sub.2, washed with saturated brine, dried over
magnesium sulfate, filtered and concentrated in vacuo. The residue
was purified by flash chromatography using 20% to 40%
EtOAc/CH.sub.2Cl.sub.2 eluent to provide 64.7 mg of the title
product. TLC; Rf=0.24, 20% EtOAc/CH.sub.2C.sub.2.
[0811] B. Compound XXII (A=Tert-butoxycarbonyl,
D'=Cyclopentylmethyl, E=3-pyridyl, Hydrochloride Salt).
[0812] A solution of 273.1 mg (0.572 mmol) of the resultant
compound of Example 149A in EtOAc (15 mL) at -20.degree. C. was
treated with anhydrous HCl gas for 10 min. The ice bath was removed
and after an additional 15 min., the reaction mixture was sparged
with nitrogen then concentrated in vacuo. To a solution of the
resulting residue in CH.sub.2Cl.sub.2 (3 mL) was added,
sequentially at ambient temperature under an atmosphere of
nitrogen, with 0.076 mL (0.437 mmol) diisopropylethylamine and 34.3
mg (0.150 mmol) of the resultant compound of Example 48A. The
mixture was stirred for 16 h and then concentrated in vacuo. The
residue was taken up in CH.sub.2Cl.sub.2 and washed with saturated
brine, dried over magnesium sulfate, filtered, and concentrated in
vacuo. The residue was purified by low pressure silica gel column
chromatography using a gradient 20% to 50% EtOAc in
CH.sub.2Cl.sub.2 eluent to yield 11.3 mg of the title product. TLC;
Rf=0.15 40% EtOAc/CH.sub.2Cl.sub.2, HPLC: Rt=13.7 min; (1H) NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 150
[0813] A. 1-Piperidinesulfonyl Chloride.
[0814] A solution of 4 g of sulfuryl chloride in acetronitrile was
treated dropwise with 861 mg of piperidine at ambient temperature
under an atmosphere of nitrogen. After complete addition, the
mixture was refluxed for 18 h, cooled to room temperature and
concentrated in vacuo to yield the title product as a red oil. TLC:
Rf=0.86, CH.sub.2Cl.sub.2. (.sup.1H)-NMR (CDCl.sub.3) consistent
with structure.
[0815] B. Compound XXII (A=Tert-butoxycarbonyl, D'=Isobutyl,
E=Piperidinyl).
[0816] A solution of 73 mg of the resultant compound of Example 39A
in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 121 mg of the
resultant compound of Example 150A and 84 mg of
N,N-diisopropylethylamine. The mixture was stirred for 14 h,
diluted with CH.sub.2Cl.sub.2, washed with saturated NaCl then
dried over MgSO.sub.4, filtered, and concentrated in vacuo. The
residue was purified by low pressure silica gel chromatography
using 5% diethyl ether/CH.sub.2Cl.sub.2 as eluent to provide 70 mg
of the title product as a white solid. TLC: Rf=0.21 (5% diethyl
ether in CH.sub.2Cl.sub.2). HPLC: Rt=17.40 min. (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
[0817] C. Compound XXII (A=H, D'=Isobutyl, E=Piperidinyl,
Hydrochloride Salt).
[0818] A solution of 70 mg of the resultant compound of Example
150B in ethyl acetate was treated at -20.degree. C. with HCl gas
for 20 min during which time the temperature was allowed to warm to
20.degree. C. Nitrogen was then bubbled through the mixture for 15
min and solvent removed in vacuo to yield a viscous oil which was
used directly for the ensuing reaction.
[0819] D. Compound 150.
[0820] A solution of the resultant compound of Example 150C in
CH.sub.2Cl.sub.2 was added, at ambient temperature under an
atmosphere of nitrogen, to a solution of 50 mg of the resultant
compound of Example 48A and 56 mg N,N-diisopropylethylamine in
CH.sub.2Cl.sub.2, the mixture was stirred for 14 h, diluted with
CH.sub.2Cl.sub.2, washed with saturated NaHCO.sub.3 and saturated
NaCl, then dried over MgSO.sub.4, filtered, and concentrated in
vacuo. The residue was purified by low pressure silica gel
chromatography using 20% diethyl ether/CH.sub.2Cl.sub.2 as eluent
to provide 16 mg of the title product as a white solid. TLC:
Rf=0.45, (0% diethyl ether/CH.sub.2Cl.sub.2, HPLC: Rt=15.00 min.
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 151
[0821] A. Compound XXII (A=Tert Butoxycarbonyl,
D'=Cyclopentylmethyl, E=4-trifluormethoxyphenyl).
[0822] A solution of 71 mg of the resultant compound of Example
114B in 4:1 CH.sub.2Cl.sub.2/saturated aqueous NaHCO.sub.3 was
treated sequentially, at ambient temperature under an atmosphere of
nitrogen, with 76 mg of 4-trifluoromethoxybenzensulfonyl chloride
and 25 mg of sodium bicarbonate. The mixture was stirred, 14 h,
diluted with CH.sub.2Cl.sub.2, washed with saturated NaCl then
dried over MgSO.sub.4, filtered, and concentrated in vacuo. The
residue was purified by low pressure silica gel chromatography
using 5% diethyl ether/CH.sub.2Cl.sub.2 as eluent to provide 92 mg
of the title product as a white solid. TLC: Rf=0.34, 5% diethyl
ether/CH.sub.2Cl.sub.2. (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
[0823] B. Compound XXII (A=H, D'=Cyclopentylmethyl,
E=4-trifluormethoxyphenyl, Hydrochloride Salt).
[0824] A solution of 92 mg of the resultant compound of Example
151A in ethyl acetate was treated at -20.degree. C. with HCl gas
for 20 min, during which time the temperature was allowed to warm
to 20.degree. C. Nitrogen was then bubbled through the mixture for
15 min and solvent removed in vacuo to yield 83 mg of white solid
which was used directly for the ensuing reaction.
[0825] C. Compound 151.
[0826] A solution of 22 mg of the resultant compound of Example
151B in CH.sub.2Cl.sub.2 was added, at ambient temperature under an
atmosphere of nitrogen, to a solution of 15 mg of the resultant
compound of Example 48A and 16 mg N,N-diisopropylethylamine in
CH.sub.2Cl.sub.2. The mixture was stirred for 60 h, diluted with
CH.sub.2Cl.sub.2, washed with saturated NaHCO.sub.3 and saturated
NaCl, then dried over MgSO.sub.4, filtered, and concentrated in
vacuo. The residue was purified by low pressure silica gel
chromatography using 20k diethyl ether/CH.sub.2Cl.sub.2 as eluent
to provide 23 mg of the title product as a white solid. TLC:
Rf=0.44, 20k diethyl ether/CH.sub.2Cl.sub.2, HPLC: Rt=16.99 min.
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 152
[0827] A. Compound XXII (A=Tert-butoxycarbonyl, D'=Isobutyl,
E=4-trifluormethoxyphenyl).
[0828] A solution of 97 mg of the resultant compound of Example 39A
in 4:1 CH.sub.2Cl.sub.2/saturated aqueous NaHCO.sub.3 was treated
sequentially, at ambient temperature under an atmosphere of
nitrogen, with 113 mg of 4-trifluoromethoxybenzenesulfonyl chloride
and 36 mg of sodium bicarbonate. The mixture was stirred for 14 h,
diluted with CH.sub.2Cl.sub.2, washed with saturated NaCl then
dried over MgSO.sub.4 filtered, and concentrated in vacuo. The
residue was purified by low pressure silica gel chromatography
using 5% diethyl ether/CH.sub.2Cl.sub.2 as eluent to provide 120 mg
of the title product as a white solid. TLC: Rf=0.34, 5% diethyl
ether/CH.sub.2Cl.sub.2, HPLC: Rt=18.54 min. (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
[0829] B. Compound XXII (A=H, D'=Isobutyl,
E=4-trifluormethoxyphenyl, Hydrochloride Salt).
[0830] A solution of 100 mg of the resultant compound of Example
152A in ethyl acetate was treated at -20.degree. C. with HCl gas
for 20 min, during which time the temperature was allowed to warm
to 20.degree. C. Nitrogen was then bubbled through the mixture for
15 min and solvent removed in vacuo to yield 89 mg of white solid
which was used directly for ensuing reaction.
[0831] C. Compound 152.
[0832] A solution of 41 mg of the resultant compound of Example
152B in CH.sub.2Cl.sub.2 was added, at ambient temperature under
the atmosphere of nigrogen, to a solution of 28 mg of the resultant
compound of Example 48A and 32 mg N,N-diisopropylethylamine in
CH.sub.2Cl.sub.2. The mixture was stirred 14 h, diluted with
CH.sub.2Cl.sub.2, washed with saturated NaHCO.sub.3 and saturated
NaCl, then dried over MgSO.sub.4, filtered, and concentrated in
vacuo. The residue was purified by low pressure silica gel
chromathography using 5% diethyl ether/CH.sub.2Cl.sub.2 as eluent
to provide 30 mg of the title product as white solid. TLC: Rf=0.08
(5% diethyl ether/CH.sub.2Cl.sub.2). HPLC: Rt=16.52 min. (1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 153
[0833] A. Compound XXII (A=Tert-butoxycarbonyl, D'=Isobutyl,
E=4-methoxyphenyl).
[0834] To a solution of the resultant compound of Example 39A (600
mg, 1.77 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added
4-methoxybenzenesulfonyl chloride (0.55 g. 2.66 mmol) followed by
the addition of a saturated solution of sodium bicarbonate (3 mL)
and 0.30 g of solid sodium bicarbonate. The mixture was stirred at
ambient temperature overnight. The solution was diluted with 200 mL
CH.sub.2Cl.sub.2 the organics were separated, dried over anhydrous
MgSO.sub.4, and the organics concentrated under reduced pressure.
The crude product was purified via medium pressure liquid
chromatography using a gradient solvent system of CH.sub.2Cl.sub.2
followed by 5:95 ether/CH.sub.2Cl.sub.2 solution to give 630 mg of
the title compound as a white solid. TLC: Rf=0.48, 3:97
methanol/CH.sub.2Cl.sub.2. (.sup.1H)-NMR (CDCl.sub.3) consistent
with structure.
[0835] B. Compound XXII (A=H, D'=Isobutyl, E=4-methoxyphenyl,
Hydrochloride Salt).
[0836] To a solution of the resultant compound of Example 153A
(0.63 g, 1.24 mmol) in EtAc (5 mL) was added 30% w/w HCl in EtOAc
(5 mL.) The mixture was stirred for 6 hours ambient temperature.
The solution was concentrated under reduced pressure to give 0.59 g
of a white solid which was used directly for subsequent reaction.
TLC Rf=0.12, 3:97 methanol/CH.sub.2Cl.sub.2.
[0837] C. Compound XXII (A=(3-pyridyl)-methyloxycarbonyl,
D'=Isobutyl, E=4-methoxyphenyl).
[0838] To a solution of the resultant compound of Example 153B (100
mg, 0.23 mmol) in CH.sub.2Cl.sub.2 (5 mL) was added triethylamine
(0.1 mL, 0.72 mmol) followed by slow addition over 3 hours of the
resultant compound of Example 82A (75 mg, 0.27 mmol) as a solution
in CH.sub.2Cl.sub.2 (5 mL). The mixture was stirred at ambient
temperature for 24 hours. The organics was concentrated under
reduced pressure and the crude product was purified via medium
pressure column chromatography using a gradient solvent system of
CH.sub.2Cl.sub.2, followed by 1:99 methanol/CH.sub.2Cl.sub.2,
followed by 3:97 methanol/CH.sub.2Cl.sub.2 as the solvent system to
give 49.3 mg of the title compound. Rf=0.33, 3:97
methanol/CH.sub.2Cl.sub.2, HPLC: Rt=13.18 min, (1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 154
[0839] Compound 154.
[0840] To a solution of the resultant compound of Example 153B (100
mg, 0.20 mmol) in CH.sub.2Cl.sub.2 (5 mL) was added triethylamine
(0.25 mL, 1.8 mmol) followed by allylchlorofornate (0.1 mL, 0.94
mmol). The mixture was stirred at ambient temperature for 24 hours.
The solution was concentrated under reduced pressure and the crude
product purified via medium pressure column chromatography using a
gradient solvent system of CH.sub.2C.sub.2, followed by 1:99
methanol/CH.sub.2Cl.sub.2 as the solvent system to give 94 mg of
the title compound. Rf=0.71, 3:97 methanol/CH.sub.2Cl.sub.2, HPLC:
Rt=16.12 min, (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 155
[0841] A. N-hydroxysuccinimidyl-1-methoxypropane-3-carbonate.
[0842] A solution of 355 mg of 2-methylene-1,3-propanediol in
acetonitrile (30 mL) was added sequentially, at ambient
temperature, 65 mg of sodium hydride and 0.25 mL iodomethane. The
mixture was stirred for 12 h and concentrated in vacuo. The residue
was then taken up in 15 mL of acetonitrile and treated
sequentially, at ambient temperature under an atmosphere of
nitrogen, with 1.3 g of N,N-disuccinimidyl carbonate and 1.6 mL of
triethylamine. After stirring for 14 h, the reaction mixture was
concentrated in vacuo and the residue was diluted CH.sub.2Cl.sub.2,
washed with saturated sodium bicarbonate solution and saturated
brine, dried over magnesium sulfate, filtered and concentrated in
vacuo. The residue was purified by silica gel chromatography with
EtOAc as eluant to give 95 mg of the title compound. (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
[0843] B. Compound 155.
[0844] A solution of 0.056 mmol of the resultant compound of
Example 40A was reacted with the resultant compound of Example 155A
in the manner described in Example 132. After concentration of the
mixture in vacuo and workup, the residue was purified by thick
layer silica gel chromatography using 7% MeOH/CH.sub.2Cl.sub.2 as
eluant followed by preparative reversed-phase C.sub.18HPLC using a
linear gradient of 35% to 100% CH.sub.3CN/H.sub.2O with 0.1 % TFA
as eluant to obtain 3.7 mg of the title compound. TLC: Rf=0.45, 7%
MeOH/CH.sub.2Cl.sub.2. HPLC: Rt=13.78 min.
EXAMPLE 156
[0845] A. 1-acetylindoline-5-sulfonyl Chloride.
[0846] A 1.02 g portion of 1-acetylindoline was treated with 2 mL
of chlorosulfonic acid at 0.degree. C. The mixture was heated at
60.degree. C. for 2 h, then treated with crushed ice, filtered and
dried to give 1.3 g of the title compound which was used directly
for subsequent reaction. TLC: Rf=0.18, 50% EtOAc/hexane.
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
[0847] B. Compound XXII (A=Tert-butoxycarbonyl,
D'=Cyclopentylmethyl, E=5-(N-acetyl)-indoline).
[0848] To a solution of 60 mg of the resultant compound of Example
114B in 15 mL of CH.sub.2Cl.sub.2 was added (5 mL) saturated
aqueous sodium bicarbonate solution, 50.0 mg sodium bicarbonate,
and 60 mg of the resultant compound of Example 156A. After 4 h, the
resulting mixture was diluted with CH.sub.2Cl.sub.2, washed with
saturated brine, dried over magnesium sulfate and filtered. The
mixture was then concentrated in vacuo to give the desired product
which was used directly for subsequent reastion. (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
[0849] C. Compound 156.
[0850] A solution of 37 mg of the resultant compound of Example
156B in EtOAc (15 mL) at 0.degree. C. was treated with anhydrous
hydrogen chloride gas for 10 min., and allowed to stand for 12 h
while warming to ambient temperature. This crude material was then
reacted with allyl chloroformate in the manner described in Example
87B. After concentration of the mixture in vacuo and workup, the
residue was purified by thick layer silica gel chromatography using
7% MeOH/CH.sub.2Cl.sub.2 as eluant followed by preparative
reversed-phase C.sub.18HPLC using a linear gradient of 35% to 100%
CH.sub.3CN/H.sub.2O with 0.1% TFA as eluant to obtain 10.5 mg of
the title compound. TLC: Rf=0.75, 10% MeOH/CH.sub.2Cl.sub.2. HPLC:
Rt=15.78 min; (.sup.1H)-NMR (CDCL.sub.3) consistent with
structure.
EXAMPLE 157
[0851] Compound 157.
[0852] A solution of 37 mg of the resultant compound of Example
156B in EtOAc (15 mL) at 0.degree. C. was treated with anhydrous
hydrogen chloride gas for 10 min., and allowed to stand for 12 h
while warming to ambient temperature. This crude material was then
reacted with the resultant compound of Example 48A in the manner
described in Example 88. After concentration of the mixture in
vacuo, the residue was purified by preparative reversed-phase
C.sub.18HPLC using a linear gradient of 35% to 100%
CH.sub.3CN/H.sub.2O with 0.1% TFA as eluant to obtain 17.9 mg of
the title compound. TLC: Rf=0.6, 10% MeOH/CH.sub.2Cl.sub.2, HPLC:
Rt=14.68 min; (.sup.1H)-NMR (CDCL.sub.3) consistent with
structure.
EXAMPLE 158
[0853] A. Compound XXII (A=Tert-butoxycarbonyl,
D'=Cyclohexylmethyl, E=H).
[0854] To a solution of compound XX (A=Boc) (5.0 mmol) in ethanol
(20 mL) was added cyclohexylmethylamine (3.25 mL, 2.83 mmol) and
the mixture was stirred for 3 hours at ambient temperature. The
solution was filtered and concentrated under reduced pressure to
give 1.49 g of a white solid which was used directly for subsequent
reaction. TLC: Rf=0.14, 3:97 methanol/CH.sub.2Cl.sub.2.
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
[0855] B. Compound XXII (A=Tert-butoxycarbonyl,
D'=Cyclohexylmethyl, E=4-methoxyphenyl).
[0856] To a solution of the resultant compound of Example 158A (400
mg, 1.06 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added
4-methoxybenzenesulfonyl chloride (0.66 g, 3.1 mmol) followed by
addition of a saturated solution of sodium bicarbonate (3 mL) and
0.18 g of solid sodium bicarbonate. The mixture was stirred at
ambient temperature overnight. The solution was diluted with 200 mL
CH.sub.2Cl.sub.2, the organics separated, dried over anhydrous
MgSO.sub.4, and the organics concentrated under reduced pressure.
The crude product was purified via medium pressure liquid
chromatography using CH.sub.2Cl.sub.2, followed by 1:99
methanol/CH.sub.2Cl.sub.2 as the solvent system to give 340 mg of
the title compound as a white solid. TLC: Rf=0.39, 3:97
methanol/CH.sub.2Cl.sub.2, (.sup.1H)-NMR (CDCL.sub.3) consistent
with structure.
[0857] C. Compound XXI (A=H, D'=Cyclohexylmethyl,
E=4-methoxyphenyl, Hydrochloride Salt).
[0858] To a solution of the resultant compound of Example 158B
(0.34 g, 0.62 mmol) in EtOAc (10 mL) was added 30% w/w HCl in EtOAc
(5 mL). The mixture was stirred for 3 hours at ambient temperature.
The solution was concentrated under reduced pressure to give 0.3 g
of a white solid which was used directly for subsequent reaction.
TLC: Rf=0.12, 3:97 methanol/CH.sub.2Cl.sub.2.
[0859] D. Compound 158.
[0860] To a solution of the resultant compound of Example 158C (100
mg, 0.21 mmol) in CH.sub.2Cl.sub.2 (8 mL) was added triethylamine
(0.2 mL, 1.44 mmol) followed by the resultant compound of Example
48A (71 mg, 0.31 mmol). The mixture was stirred at ambient
temperature for 6 hours. The solution was diluted with
CH.sub.2Cl.sub.2, (200 mL) washed with a saturated solution of
sodium bicarbonate (30 mL), the organics separated, dried over
anhydrous MgSO.sub.4 and concentrated under reduced pressure and
the crude product purified via medium pressure column
chromatography using a gradient solvent system of CH.sub.2Cl.sub.2
followed by 10:90 EtOAc/CH.sub.2Cl.sub.2 as the solvent system to
give 84.9 mg of the title compound. TLC: Rf=0.48, 3:97
methanol/CH.sub.2Cl.sub.2, HPLC: Rt=16.35 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 159
[0861] A. Compound XXII (A=Tert-butoxycarbonyl,
D'=Cyclohexylmethyl, E=4-fluorophenyl).
[0862] To a solution of the resultant compound of Example 158A (400
mg, 1.06 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added
4-fluorobenzenesulfonyl chloride (0.62 g, 3.2 mmol) followed by
addition of a saturated solution of sodium bicarbonate (3 mL) and
0.18 g of solid sodium bicarbonate. The mixture was stirred at
ambient temperature overnight. The solution was diluted with 200 mL
CH.sub.2Cl.sub.2, the organics separated, dried over anhydrous
MgSO.sub.4, and the organics concentrated under reduced pressure.
The crude product was purified via medium pressure liquid
chromatography using CH.sub.2Cl.sub.2 followed by 1:99
methanol/CH.sub.2Cl.sub.2 solution as the solvent system to give
280 mg of a white solid. TLC: Rf=0.47, 3:97
methanol/CH.sub.2Cl.sub.2, (.sup.1H)-NMR (CDCl.sub.3) consistent
with structure.
[0863] B. Compound XXII (A=H, D'=Cyclohexylmethyl,
E=4-fluorophenyl, Hydrochloride Salt).
[0864] To a solution of the resultant compound of Example 159A
(0.28 g, 0.52 mmol) was added 30% w/w HCl in EtOAc (10 mL). The
mixture was stirred for 3 hours at ambient temperature. The
solution was concentrated under reduced pressure to give 0.23 g of
a white solid which was used directly for subsequent reaction. TLC:
Rf=0.13, (3:97 methanol/CH.sub.2Cl.sub.2, (.sup.1H)-NMR
(CDCL.sub.3) consistent with structure.
[0865] C. Compound 159.
[0866] To a solution of the resultant compound of Example 159C (100
mg, 0.21 mmol) in CH.sub.2Cl.sub.2 (8 mL) was added triethylamine
(0.2 mL, 1.44 mmol) followed by the resultant compound of Example
48A (73 mg, 0.32 mmol). The mixture was stirred at ambient
temperature for 6 hours. The solution was diluted with
CH.sub.2Cl.sub.2, (200 mL) washed with saturated solution of sodium
bicarbonate (30 mL), dried over anhydrous MgSO.sub.4, the organics
concentrated under reduced pressure and the crude product purified
via medium pressure column chromatography using a gradient solvent
system of CH.sub.2Cl.sub.2, followed by 10:90
EtOAc/CH.sub.2Cl.sub.2 as the solvent system to give 54 mg of the
title compound. TLC: Rf=0.46, 3:97 methanol/CH.sub.2Cl.sub.2, HPLC:
Rt=16.48 min; (.sup.1H)-NMR (CDCL.sub.3) consistent with
structure.
EXAMPLE 160
[0867] A. Compound XXII (A=Tert-butoxycarbonyl,
D'=Cyclohexylmethyl, E=4-acetamidophenyl).
[0868] To a solution of the resultant compound of Example 158A (400
mg, 1.06 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added
4-acetamidobenzenesulfon- yl chloride (0.75 g, 3.2 mmol) followed
by addition of a saturated solution of sodium bicarbonate (3 mL)
and 0.18 g of solid sodium bicarbonate. The mixture was stirred at
ambient temperature overnight. The solution was diluted with 200 mL
CH.sub.2Cl.sub.2, the organics separated, dried over anhydrous
MgSO.sub.4, and the organics concentrated under reduced pressure.
The crude product was purified via medium pressure liquid
chromatography using CH.sub.2Cl.sub.2, followed by 1:99
methanol/CH.sub.2Cl.sub.2 and 2:98 methanol/CH.sub.2Cl.sub.2 as the
solvent system to give 290 mg of the title compound as a white
solid. TLC: Rf=0.14, 3:97 methanol/CH.sub.2Cl.sub.2, (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
[0869] B. Compound XXII (A=H, D'=Cyclohexylmethyl,
E=4-acetamidophenyl, Hydrochloride Salt).
[0870] To the resultant compound of Example 160A (0.29 g, 0.51
mmol) was added 30% w/w HCl in EtOAc (10 mL). The mixture was
stirred for 3 hours at ambient temperature. The solution was
concentrated under reduced pressure to give 0.28 g of a white solid
which was used directly for subsequent reaction. TLC: Rf=0.10, 3:97
methanol/CH.sub.2Cl.sub.2.
[0871] C. Compound 160.
[0872] To a solution of the resultant compound of Example 160B (100
mg, 0.20 mmol) in CH.sub.2Cl.sub.2 (8 mL) was added triethylamine
(0.2 mL, 1.44 mmol) followed by the resultant compound of Example
48A (67 mg, 0.30 mmol). The mixture was stirred at ambient
temperature for 6 hours. The solution was diluted with
CH.sub.2Cl.sub.2, (200 mL) washed with saturated solution of sodium
bicarbonate (30 mL), dried over anhydrous MgSO.sub.4, the organics
concentrated under reduced pressure and the crude product purified
via medium pressure column chromatography using a gradient solvent
system of CH.sub.2Cl.sub.2, followed by 10:90
EtOAc/CH.sub.2Cl.sub.2, followed by 20:80 EtOAc/CH.sub.2Cl.sub.2 as
the solvent system to give 56.8 mg of a white solid. TLC: Rf=0.17,
3:97 methanol/CH.sub.2Cl.sub.2, HPLC: Rt=14.65 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 161
[0873] A. 4-Morpholinesulfonyl Chloride.
[0874] A solution of 4.6 g of sulfuryl chloride in acetonitrile was
treated dropwise with 996 mg of morpholine at ambient temperature
under an atmosphere of nitrogen. After complete addition, the
mixture was refluxed for 16 h, cooled to room temperature, and
concentrated in vacuo to yield the title product as a red oil. TLC:
Rf=0.65 CH.sub.2Cl.sub.2. (1H)-NMR (CDCl.sub.3) consistent with
structure.
[0875] B. Compound XXII (A-tert-butoxycarbonyl, D'=Isobutyl,
E=Morpholinyl).
[0876] A solution of 98 mg of the resultant compound of Example 39A
in 4:1 CH.sub.2Cl.sub.2/saturated aqueous NaHCO.sub.3 was treated
sequentially, at ambient temperature under an atmosphere of
nitrogen, with 270 mg of the resultant compound of Example 161A and
122 mg of sodium bicarbonate. The mixture was stirred for 14 h,
diluted with CH.sub.2Cl.sub.2, dried over MgSO.sub.4, filtered, and
concentrated in vacuo. The residue was purified by low pressure
silica gel chromatography using CH.sub.2Cl.sub.2 as eluent followed
by preparative HPLC to provide 22 mg of the title product as an
oily solid. TLC: Rf=0.46, 20% diethyl ether/CH.sub.2Cl.sub.2, HPLC:
Rt=15.50 min. (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
[0877] C. Compound XXII (A=H, D'=Isobutyl, E=Morpholinyl,
Hydrochloride Salt).
[0878] A solution of 22 mg of the resultant compound of Example
161B in ethyl acetate was treated at -20.degree. C. Nitrogen was
then bubbled through the mixture for 15 min and solvent removed in
vacuo to yield an oily semi-solid mass which was used directly for
the ensuing reaction.
[0879] D. Compound 161.
[0880] A solution of the resultant compound of Example 161C in
CH.sub.2Cl.sub.2 was added, at ambient temperature under an
atmosphere of nitrogen, to a solution of 16 mg of the resultant
compound of Example 48A and 18 mg N,N-diisopropylethylamine in
CH.sub.2Cl.sub.2. The mixture was stirred for 14 h, diluted with
CH.sub.2Cl.sub.2, washed and saturated with NaHCO.sub.3 and
saturated NaCl, then dried over MgSO.sub.4, filtered, and
concentrated in vacuo. The residue was purified by preparative HPLC
to provide 21 mg of the title product as an oily solid. TLC:
Rf=0.22, 20% diethyl ether/CH.sub.2Cl.sub.2, HPLC: Rt=13.01 min.
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 162
[0881] Compound 162.
[0882] A solution of 30 mg of the resultant compound of Example
166A was deprotected with hydrogen chloride gas and the resultant
compound was reacted with the resultant compound of Example 155A in
the manner described in Example 155B. After concentration of the
mixture in vacuo and workup, the residue was purified by thick
layer silica gel chromatography using 5% MeOH/CH.sub.2Cl.sub.2 as
eluant, followed by preparative reversed-phase C.sub.18HPLC using a
linear gradient of 35% to 100% CH.sub.3CN/H.sub.2) with 0.1% TFA as
eluant to obtain 6.2 mg of the title compound. TLC: Rf=0.65, 5%
MeOH/CH.sub.2Cl.sub.2, HPLC: Rt=15.93 min .sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 163
[0883] Compound 163.
[0884] A 120.3 mg portion of the resultant compound of Example 153B
was reacted with the resultant compound of Example 82A as described
in Example 82B. After workup and concentration in vacuo, the
residue was purified by low pressure silica gel column
chromatography using 50% EtOAc in CH.sub.2Cl.sub.2 eluent, followed
by preparative reversed-phase C.sub.18HPLC using a linear gradient
of 40% to 100% acetonitrile/water for elution to obtain 44.3 mg of
the title compound. TLC: Rf=0.18, 50% EtOAc/CH.sub.2Cl.sub.2, HPLC:
Rt=13.13 min; (.sup.1H) NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 164
[0885] A. N-hydroxysuccinimidyl-(2-phenyl)ethyl Carbonate.
[0886] A solution of 306 mg of phenethyl alcohol and 535 mg of
N,N'-disuccinimidyl carbonate in acetonitrile was treated, at
ambient temperature under an atmosphere of nitrogen, with 810 mg of
N,N-diisopropylethylamine. The mixture was stirred for 60 h and
concentrated in vacuo. The residue was taken up in ethyl acetate
and washed with saturated NaHCO.sub.3, saturated NaCl, then dried
over MgSO.sub.4, filtered, and concentrated in vacuo to yield the
title product as a yellow oil. TLC: Rf=0.40 (5% methanol in
CH.sub.2Cl.sub.2). (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
[0887] B. Compound 164.
[0888] A solution of 81 mg of the resultant compound of Example
164A in CH.sub.2Cl.sub.2 was added, at ambient temperature under an
atmosphere of nitrogen, to a solution of 41 mg of the resultant
compound of example 40a and 45 mg N,N-diisopropylethylamine in
CH.sub.2Cl.sub.2. The mixture was stirred for 4 h, diluted with
CH.sub.2Cl.sub.2, washed with saturated NaHCO.sub.3 and saturated
NaCl, then dried over MgSO.sub.4, filtered, and concentrated in
vacuo. The residue was subjected to preparative HPLC to yield 18 mg
of the title product. TLC: Rf=0.83 (5:10:85
NH.sub.4H/CH.sub.3OH/CH.sub.2Cl.sub.2). HPLC: Rt=15.78 min.
.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 165
[0889] Compound 165.
[0890] A solution of 36 mg of the resultant compound of Example 51D
in 4:1 CH.sub.2Cl.sub.2/saturated aqueous NaHCO.sub.3 was treated
sequentially, at ambient temperature under an atmosphere of
nitrogen, with 20 mg of p-toluenesulfonyl chloride and 18 mg of
sodium bicarbonate. The mixture was stirred for 3 h, diluted with
CH.sub.2Cl.sub.2, washed with saturated NaCl then dried over
MgSO.sub.4, filtered, and concentrated in vacuo. The residue was
purified by low pressure silica gel chromatography using 5% diethyl
ether/CH.sub.2Cl.sub.2 as eluent to provide 38 mg of the title
product as a white solid. TLC: Rf=0.15, 5% diethyl
ether/CH.sub.2Cl.sub.2, HPLC: Rt=15.27 min. (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 166
[0891] A. Compound XXII (A=Tert-butoxycarbonyl,
D'=Cyclopentylmethyl, E=4-methoxyphenyl).
[0892] To a solution of the resultant compound of Example 114B (1.8
g, 4.96 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added
4-methoxylbenzensulfonyl chloride (2.10 g, 9.93 mmol), followed by
addition of a saturated solution of sodium bicarbonate (3 mL) and
0.83 g of solid sodium bicarbonate. The mixture was stirred at
ambient temperature for 24 hours. The solution was diluted with 200
mL CH.sub.2Cl.sub.2 the organics were separated, dried over
anhydrous MgSO.sub.4, and concentrated under reduced pressure. The
crude product was purified via medium pressure liquid
chromatography using CH.sub.2Cl.sub.2, followed by 1:99
methanol/CH.sub.2Cl.sub.2 followed by 2:98
methanol/CH.sub.2Cl.sub.2 as the solvent system to give 1.49 g of
the title compound as a white solid. TLC: Rf=0.37, 3:97
methanol/CH.sub.2Cl.sub.2; (.sup.1H)-NMR (CDCl.sub.3) consistent
with structure.
[0893] B. Compound XXII (A=H, D=Cyclopentylmethyl,
E=4-hydroxyphenyl).
[0894] A solution of the resultant compound of Example 166A (1.11
g, 2.08 mmol) in CH.sub.2Cl.sub.2 (20 mL) was added to a solution
of boron tribromide in CH.sub.2Cl.sub.2 (1.0 M, 10.4 mL). The
mixture was stirred at ambient temperature for 24 hours. The
solution was poured onto 40 mL of a saturated solution of sodium
bicarbonate. The aqueous layer was extracted with 250 mL
CH.sub.2Cl.sub.2 followed by extraction with 250 mL EtOAc. The
combined organics were dried over anhydrous MgSO.sub.4,
concentrated under reduced pressure and the crude product purified
via medium pressure column chromatography using a gradient solvent
system of CH.sub.2Cl.sub.2, followed by 1:99
methanol/CH.sub.2Cl.sub.2, followed by 9:98
methanol/CH.sub.2Cl.sub.2, followed by a 1:5:95 concentrated
NH.sub.4OH/methanol/CH.sub.2Cl.sub.2 solution as the solvent system
to give 0.38 g of the title compound. TLC: Rf=0.18, 3:97
methanol/CH.sub.2Cl.sub.2, (.sup.1H)-NMR (CDCl.sub.3) consistent
with structure.
[0895] C. Compound 166.
[0896] To a solution of the resultant compound of Example 166B (300
mg, 0.69 mmol) in CH.sub.2Cl.sub.2 (5 mL) was added triethylamine
(0.12 mL, 8.6 mmol), followed by slow addition over 3 hours of the
resultant compound of Example 82A (0.21 g, 0.77 mmol) as a solution
in CH.sub.2Cl.sub.2 (5 mL). The mixture was stirred at ambient
temperature for 24 hours. The solution was diluted with 250 mL
CH.sub.2Cl.sub.2, washed with water, dried over anhydrous
MgSO.sub.4, and the organics concentrated under reduced pressure.
The crude product was purified via medium pressure column
chromatography using a gradient solvent system of CH.sub.2Cl.sub.2
followed by 1:99 methanol/CH.sub.2Cl.sub.2, followed by 2:98
methanol/CH.sub.2Cl.sub.2 as the solvent system to give 110 mg of a
white solid. TLC: Rf=0.14 (3:97 methanol/CH.sub.2Cl.sub.2), HPLC:
Rt=12.69 min, 1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 167
[0897] Compound 167.
[0898] A solution of 102 mg of the resultant compound of Example
51D in 4:1 CH.sub.2Cl.sub.2/saturated aqueous NaHCO.sub.3 was
treated sequentially, at ambient temperature under an atmosphere of
nitrogen, with 65 mg of p-nitrobenzenesulfonyl chloride and 51 mg
of sodium bicarbonate. The mixture was stirred for 14 h, diluted
with CH.sub.2Cl.sub.2, washed with saturated NaCl, then dried over
MgSO.sub.4, filtered, and concentrated in vacuo. The residue was
purified by low pressure silica gel chromatography using 20%
diethyl ether/CH.sub.2Cl.sub.2 as eluent to provide 124 mg of the
title product as a white solid. TLC: Rf=0.36, 20% diethyl
ether/CH.sub.2Cl.sub.2, HPLC: Rt=15.15 min. (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 168
[0899] Compound 168.
[0900] A solution of 124 mg of the resultant compound of Example
167 in ethyl acetate was treated, at ambient temperature, with 13
mg of 10% palladium on carbon. The mixture was stirred for 14 h
under an atmosphere of hydrogen, filtered through a pad of Celite
filter agent, and concentrated in vacuo. The residue was subjected
to preparative HPLC to yield 82 mg of the title product as a white
solid. TLC: Rf=0.10, 20% ether/CH.sub.2Cl.sub.2, HPLC: Rt=13.16
min. (.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 169
[0901] Compound 169.
[0902] To a solution of the resultant compound of Example 166B (80
mg, 0.18 mmol) in CH.sub.2Cl.sub.2 (15 mL) was added a saturated
solution of sodium bicarbonate (5 mL) followed by the addition of
the resultant compound of Example 48A (55 mg, 0.24 mmol). The
mixture was stirred at ambient temperature for 5 hours. The
solution was diluted with 200 mL CH.sub.2Cl.sub.2, the organics
separated, dried over anhydrous MgSO.sub.4, and concentrated under
reduced pressure. The crude product was purified via medium
pressure liquid chromatography using CH.sub.2Cl.sub.2, followed by
1:99 methanol/CH.sub.2Cl.sub.2 as the solvent system to give 56 mg
of the title compound as a white solid. TLC: Rf=0.24, 3:97
methanol/CH.sub.2Cl.sub.2, HPLC: Rt=14.29 min. (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 170
[0903] A. Compound XXII (A=Tert-butoxycarbonyl,
D'=Cyclopentylmethyl, E=4,nitrophenyl).
[0904] To a solution of the resultant compound of Example 114B (250
mg, 0.69 mmol) in CH.sub.2Cl.sub.2 (15 mL) was added a saturated
solution of sodium bicarbonate (5 mL) followed by solid sodium
bicarbonate (0.12 g, 1.37 mmol) and 4-nitrobenzensulfonyl chloride
(200 mg, 0.9 mmol). The mixture was stirred at ambient temperature
for 24 hours. The solution was diluted with 200 mL
CH.sub.2Cl.sub.2, the organics separated, dried over anhydrous
MgSO.sub.4, and concentrated under reduced pressure. The crude
product was purified via medium pressure liquid chromatography
using a gradient solvent system of CH.sub.2Cl.sub.2 followed by
1:99 methanol/CH.sub.2Cl.sub.2 to give 360 mg of the title compound
as an orange solid. TLC: Rf=0.45, 3:97 methanol/CH.sub.2Cl.sub.2.
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
[0905] B. Compound XXII (A=H, D'=Cyclopentylmethyl,
E-4-nitrophenyl, Hydrochloride Salt).
[0906] To the resultant compound of Example 170A (360 mg, 0.66
mmol) was added 10% w/w HCl in EtOAc (15 mL). The mixture was
stirred for 3 hours at ambient temperature. The solution was
concentrated under reduced pressure to give 310 mg of the title
compound as an orange solid which was used directly for subsequent
reaction. TLC: Rf=0.70, 1:10:90
NH.sub.4OH/methanol/CH.sub.2Cl.sub.2.
[0907] C. Compound 170.
[0908] To a solution of the resultant compound of Example 170B (310
mg, 0.64 mmol) in CH.sub.2Cl.sub.2 (15 mL) was added a saturated
solution of sodium bicarbonate (5 mL) followed by the addition of
solid sodium bicarbonate (0.11 g, 1.3 mmol) and the resultant
compound of Example 48A (0.18 g, 0.77 mmol). The mixture was
stirred at ambient temperature for 24 hours. The solution was
diluted with 150 mL CH.sub.2Cl.sub.2, the organics separated, dried
over anhydrous MgSO.sub.4, and concentrated under reduced pressure.
The crude product was purified via medium pressure liquid
chromatography using CH.sub.2Cl.sub.2, followed by 1:99
methanol/CH.sub.2Cl.sub.2 as the solvent system to give 0.32 g of
the title compound as a white solid. TLC: Rf=0.28, 3:97
methanol/CH.sub.2Cl.sub.2, HPLC: Rt=16.06 min, .sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 171
[0909] Compound 171.
[0910] A solution of the resultant compound of Example 170C (0.19
g, 0.34 mmol) in EtOAc (10 mL) was treated at ambient temperature
with 50 mg of 10% palladium on carbon and hydrogenated for 72 hours
under a slight positive pressure of hydrogen. The mixture was
filtered and concentrated in vacuo and the crude product purified
via medium pressure liquid chromatography using CH.sub.2Cl.sub.2,
followed by 1:99 methanol/CH.sub.2Cl.sub.2, followed by 3:97
methanol/CH.sub.2Cl.sub.2, followed by 10:90
methanol/CH.sub.2Cl.sub.2 as the solvent system to give 97 mg of
the title compound as a white solid. TLC: Rf=0.25, 3:97
methanol/CH.sub.2Cl.sub.2, HPLC: Rt=14.28 min, (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 172
[0911] A. Compound XXII (A=Tert-butoxycarbonyl,
D=Cyclopentylmethyl, E=2,4-dinitrophenyl).
[0912] To a solution of the resultant compound of Example 114B (500
mg, 1.38 mmol) in CH.sub.2Cl.sub.2 (15 mL) was added a saturated
solution of sodium bicarbonate (5 mL) followed by solid sodium
bicarbonate (0.23 g, 2.76 mmol) and 2,4-dinitrobenzenesulfonyl
chloride (440 mg, 1.65 mmol). The mixture was stirred at ambient
temperature for 2 hours. The solution was diluted with 200 mL
CH.sub.2Cl.sub.2, the organics separated, dried over anhydrous
MgSO.sub.4, and concentrated under reduced pressure. The crude
product was purified via medium pressure liquid chromatography
using a gradient solvent system of CH.sub.2Cl.sub.2, followed by
1:99 methanol/CH.sub.2Cl.sub.2 to give 700 mg of the title compound
as a brown solid. TLC: Rf=0.48, 3:97 methanol/CH.sub.2Cl.sub.2,
(.sup.1H)-NMR (CDCl.sub.3), consistent with structure.
[0913] B. Compound XXII (A=H, D=Cyclopentylmethyl,
E-2,4-dinitrophenyl, Hydrochloride Salt).
[0914] To a the resultant compound of Example 172A (700 mg, 1.18
mmol) was added 10% w/w HCl in EtOAc (20 mL). The mixture was
stirred for 3 hours at ambient temperature. The solution was
concentrated under reduced pressure to give 590 mg of the title
compound as a brown solid which was used without subsequent
purification. TLC: Rf=0.55, 1:10:90
NH.sub.4OH/methanol/CH.sub.2Cl.sub.2.
[0915] C. Compound 172.
[0916] To a solution of the resultant compound of 172B (590 mg,
1.11 mmol) in CH.sub.2Cl.sub.2 (15 mL) was added a saturated
solution of sodium bicarbonate (5 mL), followed by solid sodium
bicarbonate (0.19 g, 2.2 mmol) and the resultant compound of
Example 48A (0.31 g, 1.3 mmol). The mixture was stirred at ambient
temperature for 24 hours. The solution was diluted with 150 mL
CH.sub.2Cl.sub.2, the organics separated, dried over anhydrous
MgSO.sub.4, and the organics concentrated under reduced pressure.
The crude product was purified via medium pressure liquid
chromatography using a CH.sub.3OH/CH.sub.2Cl.sub.2 gradient as
eluant, to yield the product as 0.59 g of a white solid. HPLC:
Rt=16.36 min, .sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 173
[0917] Compound 173.
[0918] A solution of the resultant compound of Example 172C (0.20
g, 0.33 mmol) in EtOAc (10 mL) was treated under ambient
temperature with 50 mg of 10% palladium on carbon and hydrogenated
for 72 hours under a slight positive pressure of hydrogen. The
mixture was filtered and concentrated in vacuo and the crude
product purified via medium pressure liquid chromatography using
CH.sub.2Cl.sub.2, followed by 1:99 methanol/CH.sub.2Cl.sub.2, 3:97
methanol/CH.sub.2Cl.sub.2, and 10:90 methanol/CH.sub.2Cl.sub.2 as
the solvent system to give 120.2 mg of the title compound as a
light brown solid. TLC: Rf=0.17, 3:97 methanol/CH.sub.2Cl.sub.2,
HPLC: Rt=13.47 min, (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 174
[0919] A. 4-Benzyloxybenzenesulfonyl Chloride.
[0920] To 0.87 g of dimethylformamide, at 0.degree. C. under an
atmosphere of nitrogen, was added 1.61 g of sulfuryl chloride. The
mixture was stirred for 15 min and treated with 2.00 g of benzyl
phenyl ether. The mixture was then heated at 100.degree. C. for 1.5
h, cooled to about 40.degree. C., poured onto ice, extracted with
CH.sub.2Cl.sub.2, dried over MgSO.sub.4, filtered, and concentrated
in vacuo. The residue was purified by low pressure silica gel
chromatography using 10% ethyl acetate in hexane as eluent to
provide 0.78 g of the title product as a white solid. TLC: Rf=0.46,
10% ethyl acetate in hexane. (.sup.1H)-NMR (CDCl.sub.3) consistent
with structure.
[0921] B. Compound 174.
[0922] A solution of 30 mg of the resultant compound of Example 51D
in 4:1 CH.sub.2Cl.sub.2/saturated aqueous NaHCO.sub.3 was treated
sequentially, at ambient temperature under an atmosphere of
nitrogen, with 24 mg of the resultant compound of Example 174A and
18 mg of sodium bicarbonate. The mixture was stirred for 14 h,
diluted with CH.sub.2Cl.sub.2, washed with saturated NaCl then
dried over MgSO.sub.4, filtered, and concentrated in vacuo. The
residue was purified by low pressure silica gel chromatography
using 20k diethyl ether/CH.sub.2Cl.sub.2 as the eluent to provide
14 mg of the title product as a white solid. TLC: Rf=0.43, 20%
diethyl ether/CH.sub.2Cl.sub.2, HPLC: Rt=17.01 min. (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 175
[0923] Compound 175.
[0924] A solution of 11 mg of the resultant compound Example 174B
in ethyl acetate was treated at ambient temperature, with 2 mg of
10% palladium on carbon. The mixture was stirred for 14 h under an
atmosphere of hydrogen, filtered through a pad of Celite filter
agent, filtered, and concentrated in vacuo. The residue was
purified by low pressure silica gel chromatography using 10%
methanol in CH.sub.2Cl.sub.2 as the eluent to provide 9 mg of the
title product as a white solid. TLC: Rf=0.38, 10% methanol in
CH.sub.2Cl.sub.2. HPLC: Rt=13.37 min. (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 176
[0925] A. 1,3-Benzodioxole-5-sulfonyl Chloride.
[0926] To 3.50 g of dimethylformamide, at 0.degree. C. under an
atmosphere of nitrogen, was added 6.47 g of sulfuryl chloride. The
mixture was stirred 15 min and treated with 5.32 g of
1,3-benxodioxole. The mixture was then heated at 120.degree. C. for
45 min, cooled to about 40.degree. C., poured onto ice, extracted
with CH.sub.2Cl.sub.2, dried over MgSO.sub.4, filtered, and
concentrated in vacuo. The residue was purified by low pressure
silica gel chromatography using 40% CH.sub.2Cl.sub.2 in hexane as
eluent to provide 2.70 g of the title product as a yellow solid.
TLC: Rf=0.37, 40% CH.sub.2Cl.sub.2 in hexane. (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
[0927] B. Compound XXII (A=Tert-butoxy, D'=Isobutyl,
E=3,4-benzodioxole).
[0928] A solution of 49 mg of the resultant compound of Example 39A
in 4:1 CH.sub.2Cl.sub.2/saturated aqueous NaHCO.sub.3 was treated
sequentially, at ambient temperature under an atmosphere of
nitrogen, with 45 mg of the resultant comnpound of Example 176A and
28 mg of sodium bicarbonate. The mixture was stirred for 14 h,
diluted with CH.sub.2Cl.sub.2, washed with saturated NaCl then
dried over MgSO.sub.4, filtered, and concentrated in vacuo. The
residue was purified by low pressure silica gel chromatography
using 20% diethyl ether/CH.sub.2Cl.sub.2 as the eluent to provide
71 mg of the title product as a waxy solid. TLC: Rf=0.65, 20%
diethyl ether/CH.sub.2Cl.sub.2. (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
[0929] C. Compound XXII (A=H, D'=Isobutyl, E=3,4-benzodioxole,
Hydrochloride Salt).
[0930] A solution of 71 mg of the resultant compound of Example
176B in ethyl acetate was treated at -20.degree. C. with HCl gas.
The HCl was bubbled through the mixture for 20 min over which time
the temperature was allowed to warm to 20.degree. C. Nitrogen was
then bubbled through the mixture for 15 min and solvent removed in
vacuo to yield 66 mg of the title product as a white solid which
was used directly in subsequent reactions.
[0931] D. Compound 176.
[0932] A solution of 18 mg of the resultant compound of Example
176C in CH.sub.2Cl.sub.2 was added, at ambient temperature under an
atmosphere of nitrogen, to a solution of 13 mg of the resultant
compound of Example 48A and 14 mg N,N-diisopropylethylamine in
CH.sub.2Cl.sub.2. The mixture was stirred for 16 h, diluted with
CH.sub.2Cl.sub.2, washed with saturated NaHCO.sub.3 and saturated
NaCl, then dried over MgSO.sub.4, filtered, and concentrated in
vacuo. The residue was purified by low pressure silica gel
chromatography using 5% diethyl ether/CH.sub.2Cl.sub.2 as the
eluent to provide 9 mg of the title product as a white solid. TLC:
Rf=0.14, 5% diethyl ether/CH.sub.2Cl.sub.2. HPLC: Rt=15.52 min;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 177
[0933] A. (4-Methoxyphenyl)-methyl-4-nitrophenyl Carbonate.
[0934] To a solution of 1.50 g of p-nitrophenyl chloroformate in 30
mL of CH.sub.2Cl.sub.2 at 0.degree. C. was added sequentially, 0.77
mL of 4-methoxybenzyl alcohol and 0.82 mL of 4-methyl morpholine.
After stirring for a half hour at ambient temperature, the
resulting mixture was diluted with CH.sub.2Cl.sub.2, washed with
water, brine, dried over magnesium sulfate, filtered and
concentrated in vacuo to yield a pale yellow solid which was
triturated with CH.sub.2Cl.sub.2/hexane and filtered to yield 1.51
g of the title compound. TLC: Rf=0.40, 20% EtOAc/hexane.
[0935] B. Compound 177.
[0936] To a solution of 96.7 mg of the resultant compound of
Example 141A in 2 mL of CH.sub.2Cl.sub.2 was added sequentially, 90
.mu.L of diisopropylethylamine and 81.3 mg of the resultant
compound of Example 178A. After stirring for 24 hours, the mixture
was diluted with CH.sub.2Cl.sub.2, washed with water and brine,
dried over magnesium sulfate, filtered and concentrated in vacuo.
The residue was purified by preparative thin layer chromatography
using 5% methanol in CH.sub.2Cl.sub.2 eluent to yield 104.8 mg of
the title compound. TLC: Rf=0.4, 20% EtOAc/hexane, HPLC: Rt=17.66
min, (.sup.1H)NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 178
[0937] A. (3-Methoxyphenyl)-methyl-4-nitrophenyl Carbonate.
[0938] Prepared by the same route as described for Example 177A,
except 3-methoxybenzyl alcohol was utilized for reaction with
p-nitrophenyl chloroformate to yield the title compound as a pale
yellow solid. TLC: Rf=0.40, 20% EtOAc/hexane.
[0939] B. Compound 178.
[0940] To a solution of 97.8 mg of the resultant compound of
Example 141A in 2 mL of CH.sub.2Cl.sub.2 was added sequentially, 91
.mu.L of disopropylethylamine and 82.2 mg of the resultant compound
of Example 178A. After stirring for 24 hours, the mixture was
diluted with CH.sub.2Cl.sub.2, washed with water and brine, dried
over magnesium sulfate, filtered and concentrated in vacuo. The
residue was purified by preparative thin layer chromatography using
5% methanol in CH.sub.2Cl.sub.2 eluent to yield 25.7 mg of the
title compound. TLC: Rf=0.4, 20% EtOAc/hexane, HPLC: Rt=17.75 min.
(.sup.1H)NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 179
[0941] A. (2-Methoxyphenyl)-methyl-4-nitrophenyl Carbonate.
[0942] Prepared by the same route as described for Example 177A,
except 2-methoxybenzyl alcohol was utilized for reaction with
p-nitrophenyl chloroformate to yield the title compound as a pale
yellow solid. TLC: Rf=0.40, 20% EtOAc/hexane.
[0943] B. Compound 179.
[0944] To a solution of 97.8 mg of the resultant compound of
Example 141A in 2 mL of CH.sub.2Cl.sub.2 was added sequentially, 99
.mu.L of diisoprophylethylamine and 89.2 mg of the resultant
compound of Example 179A. After stirring for 24 hours the mixture
was diluted with CH.sub.2Cl.sub.2, washed with water and brine,
dried over magnesium sulfate, filtered and concentrated in vacuo.
The residue was purified by preparative thin layer chromatography
using 5% methanol in CH.sub.2Cl.sub.2 eluent to yield 107.0 mg of
the title compound. TLC: Rf=0.4, 20% EtOAc/hexane, HPLC: Rt=17.58
min. (.sup.1H)NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 180
[0945] A. 2,3-Dihydrobenzofuran-5-sulfonyl Chloride.
[0946] To. 3.35 g of dimethylformamide, at 0.degree. C. under an
atmosphere of nitrogen, added 6.18 g of sulfuryl chloride. The
mixture was stirred 15 min and treated with 4.69 g of
2,3-dihydrobenzofuran. The mixture was then heated at 100.degree.
C. for 1.5 h, cooled to about 40.degree. C., poured onto ice,
extracted with CH.sub.2Cl.sub.2, dried over MgSO.sub.4, filtered,
and concentrated in vacuo. The residue was taken up in ethyl
acetate, cooled to 5.degree. C. for 16 h, and the resultant pink
crystals collected by vacuum filtration to provide 6.12 g of the
title product. TLC: Rf=0.41, 10% ethyl acetate in hexane.
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
[0947] B. Compound 180.
[0948] A solution of 32 mg of the resultant compound of Example
140D in 4:1 CH.sub.2Cl.sub.2/saturated aqueous NaHCO.sub.3 was
treated sequentially, at ambient temperature under an atmosphere of
nitrogen, with 22 mg of the resultant compound of Example 180A and
18 mg of sodium bicarbonate. The mixture was stirred 14 h, diluted
with CH.sub.2Cl.sub.2 washed with saturated NaCl then dried over
MgSO.sub.4, filtered, and concentrated in vacuo. The residue was
purified by low pressure silica gel chromatography using 20%
diethyl ether/CH.sub.2Cl.sub.2 as eluent to provide 20 mg of the
title product as a white solid. TLC: Rf=0.52, 20% diethyl
ether/CH.sub.2Cl.sub.2, HPLC: Rt=15.49 min .sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 181
[0949] Compound 181.
[0950] A solution of the resultant compound of Example 140D (150
mg, 0.4 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added a saturated
solution of sodium bicarbonate (5 mL) followed by solid sodium
bicarbonate (0.1 g, 1.2 mmol) and 4-cyanobenzensulfonyl chloride
(0.1 g, 0.48 mmol). The mixture was stirred at ambient temperature
for 4 hours. The solution was diluted with 200 mL CH.sub.2Cl.sub.2,
the organics separated, dried over anhydrous MgSO.sub.4, and the
organics concentrated under reduced pressure. The crude product was
purified via medium pressure liquid chromatography using
CH.sub.2Cl.sub.2, followed by 1:99 methanol/CH.sub.2Cl.sub.2
solution as the solvent system to give 0.19 g (86% yield) of the
title compound as a white solid. TLC: Rf=0.40, 3:97
methanol/CH.sub.2Cl.sub.2, HPLC: Rt=15.02 min, (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 182
[0951] Compound 182.
[0952] This compound was prepared from the resultant compound of
Example 114D and the resultant compound of Example 48A in the same
manner described in Example 88. After workup and purification by
preparative reversed-phase C.sub.18HPLC using a linear gradient of
35% to 100% CH.sub.3CN/H.sub.2O with 0.1% TFA as eluant, 32.8 mg of
the title compound was obtained. TLC: Rf=0.25, 4%
MeOH/CH.sub.2Cl.sub.2, HPLC: Rt=16.06 min; (.sup.1H)NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 183
[0953] Compound 183.
[0954] This compound was prepared from the resultant compound of
Example 84 by treatment with hydrogen chloride gas and subsequent
reaction with the resultant compound of Example 48A in the manner
described in Example 132. After workup and purification by
crystallization from EtOAc, 33.0 mg of the title compound was
obtained as a white solid. TLC: Rf=0.25, 4% MeOH/CH.sub.2Cl.sub.2,
HPLC: Rt=17.71 min; (.sup.1H)-NMR (CDCL.sub.3) consistent with
structure.
EXAMPLE 184
[0955] A.
(N-tert-butoxycarbonyl)-(R)-3-pyrrolidinyl-N-hydroxysuccinimidyl
Carbonate.
[0956] To a solution of 1.0 g of (R)-3-hydroxypyrrolidine in
tetrahydrofuran (50 mL) was added sequentially, at ambient
temperature, 3.75 g of di-tert-butyl dicarboante and 1 mL of 2N
sodium hydroxide. The mixture was stirred for 1 hour, filtered and
concentrated in vacuo. The resultant compound was reacted with
N,N-disuccinimidyl carbonate in the manner described in Example
155A. Workup and purification by thick layer silica gel
chromatography using an EtOAc eluent yielded the title compound as
a white solid; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
[0957] B. Compound 184.
[0958] A solution of 350 mg of the resultant compound of Example
166A was deprotected with hydrogen chloride gas and the resultant
compound was reacted with the resultant compound of Example 184A in
the manner described in Example 88. After concentration of the
mixture in vacuo and workup, the residue was purified by thick
layer silica gel chromatography using 7% MeOH/CH.sub.2Cl.sub.2 as
eluant, to obtain 120 mg of the title compound. TLC: Rf=0.45, 5%
MeOH/CH.sub.2Cl.sub.2, HPLC: Rt=16.97 min; (1H)-NMR (CDCl.sub.3)
consistent with structure.
EXAMPLE 185
[0959] Compound 185.
[0960] A solution of 120 mg of the resultant compound of Example
184B in EtOAc (25 mL) at 0.degree. C. was treated with anhydrous
hydrogen chloride gas for 10 min., and allowed to stand for 12 h
while warming to ambient temperature. Concentration in vacuo
yielded 110 mg of the title compound. TLC: Rf=0.35, 10% MeOH/89%
CH.sub.2Cl.sub.2/1% NH.sub.4OH. HPLC: Rt=13.72 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 186
[0961] A. Compound XXX ((syn, anti)-OH, A=Carbobenzyloxy,
R.sup.3=(s)-sec-butyl, R.sup.3=H, D'-benzyl,
A'=Tert-butoxycarbonyl).
[0962] A solution of 1.37 g of the resultant compound of Example 1B
in 150 mL of methylene chloride was treated with 1.03 of Cbz-Ile,
523 mg of HOBT.H.sub.2O, and 742 mg of EDC. The mixture was stirred
for 18 h, then diluted with 3 volumes of diethyl ether and washed
sequentially with water, saturated NaHCO.sub.3 solution, 10%
KHSO.sub.4 solution, and brine. After drying over MgSO.sub.4 and
concentrating in vacuo, the residue was purified by chromatography
on a silica gel column using a gradient of 1% to 1.5% MeOH in
CH.sub.2Cl.sub.2 as eluant to yield 2.10 g of the title compound as
a white foam. TLC: Rf=0.51, 5% methanol/CH.sub.2Cl.sub.2.
[0963] B. Compound XXX ((syn, anti)-OH, A=Carbobenzyloxy,
R.sup.3=(S)-sec-butyl, R.sup.3=H, D'=Benzyl, A'=H), Hydrochloride
Salt.
[0964] A solution of 650 mg of the resultant compound of Example
12A in 12 mL of ethyl acetate was cooled in an ice/water bath and
treated with a slow stream of HCl gas for approximately 6 min with
vigorous stirring. The mixture was capped and stirred for an
additional 10 min, then purged with a stream of nitrogen for 15
minutes and concentrated in vacuo to yield a white solid which was
used without subsequent purification. TLC: Rf=0.18, 95:5:0.5
CH.sub.2Cl.sub.2/methanol/concentrated NH.sub.4OH.
[0965] C. Compound 186.
[0966] A solution of 20 mg of the resultant compound of Example
186B in 0.8 mL of methylene chloride was cooled in ice/methanol
(approximately 15.degree. C.), then treated with 13.8 .mu.L of DIEA
followed by 7.6 mg of a-toluene sulfonyl chloride. The mixture was
stirred for 15 h, warming slowly to ambient temperature. The
mixture was concentrated to a small volume, applied to a 0.5 mm
thick prep plate and eluted with 3.5% MeOH/CH.sub.2Cl.sub.2. The
band containing the desired diastereomer was isolated and eluted
with 8% MeOH/CH.sub.2Cl.sub.2 to yield 4.8 mg of the title
compound. TLC: Rf=0.42, 15% diethyl ether/CH.sub.2Cl.sub.2, HPLC:
Rt=17.81 min. NMR (CDCl.sub.3): 0.78 (dd, 6H) 0.84 (m, 1H) 1.07,
(m, 1H) 1.76-1.86 (m, 2H) 2.72 (m, 2H); 3.14 (s, 2H); 3.49 (dd,
1H); 3.87 (dd, 1H); 3.58 (m, 1H); 4.01 (d, 1H); 4.14, (d, 1H);
4.26, (d, 1H); 4.35, (d, 1H); 4.90, (m, 1H); 5.08, (s, 2H); 5.97,
(d, 1H), 7.08, (d, 2H); 717, (t, 1H); 7.20-7.40, (m, 17H).
EXAMPLE 187
[0967] Compound 187.
[0968] 100 mg of the resulting compound 54A was treated with 1 mL
of 90% aqueous TFA and allowed to stand for 12 h. The mixture was
concentrated in vacuo and the residue taken up in 10 mL of dry
CH.sub.2Cl.sub.2, treated with 65 mg of N-Cbz-L-isoleucine (0.235
mmol), 50 .mu.L of DIEA (0.27 mmoles), 30 mg of HOBt (0.22 mmoles),
and 42 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride (0.22 mmoles). The mixture was stirred for 3 h, then
diluted with in CH.sub.2Cl.sub.2 and washed sequentially with
water, saturated NaHCO.sub.3 solution, and brine. After drying over
MgSO.sub.4 and concentrating in vacuo, the mixture was purified by
chromatography on a silica gel column using 5% CH.sub.3OH in
CH.sub.2Cl.sub.2 as eluent to yield the title compound, a portion
which was purified by preparative reversed-phase C.sub.18HPLC using
a linear gradient of 35% to 100% CH.sub.3CN/H.sub.2O with 0.1W TFA
for elution to obtain 36.0 mg 99.0% pure compound. TLC: Rf=0.25, 5k
CH.sub.3OH in CH.sub.2Cl.sub.2, HPLC: Rt=16.45 min; H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 188
[0969] Compound 188.
[0970] A solution of 51 mg of the resulting compound of Example
187A in 15 mL of methanol was hydrogenated under a slight positive
pressure of hydrogen in the presence of 10 mg of 10% Pd(OH).sub.2
for 14 h. After filtering and concentrating in vacuo, the crude
mixture was taken up into 10 mL CH.sub.2Cl.sub.2 and treated with
0.203 mL of DIEA and 19.0 mg of 2-quinoxaloyl chloride. The mixture
was stirred for 6 h, then diluted with CH.sub.2Cl.sub.2 and washed
with water. After dryuing over MgSO.sub.4 and concentrating in
vacuo, a portion of the mixture was purified by preparative
reversed-phase Cl.sub.8HPLC using a linear gradient of 35% to 100%
CH.sub.3CN/H.sub.2O with 0.1% TFA for elution to obtain 2.1 mg of
the title compound. TLC: Rf=0.25, 6% Ch.sub.3CN/H.sub.2O with 0.1%
TFA for elution to obtain 2.1 mg of the title compound. TLC:
Rf=0.25, 6% CH.sub.3OH in CH.sub.2Cl.sub.2. HPLC: Rt=16.21 min;
(.sup.1H)-NMR (CDCl.sub.3) consistent with structure.
EXAMPLE 189
[0971] A. Compound XXII (D'=Isobutyl, A=H, E=4-acetamidophenyl,
Trifluoroacetate Salt).
[0972] To a solution of 89.3 mg. (0.167 mmol) of the resultant
compound of Example 39B in CH.sub.2Cl.sub.2 (1 mL) at 0.degree. to
5.degree. C. was added trifluoromethanesulfonic acid (1 mL). After
stirring for 0.5 h the resultant mixture was concentrated in vacuo
and the resulting yellow gum used without subsequent
purification.
[0973] B. Compound 189.
[0974] A solution of the resultant compound of Example 189A (0.167
mmol) in CH.sub.2Cl.sub.2 was treated sequentially, at ambient
temperature under an atmosphere of nitrogen, with 44.2 mg (0.217
mmol) of N-Boc-.alpha.-aminoisobutyric acid, 0.044 mL (0.251 mmol)
diisopropylethylamine, 27.1 mg (0.201 mmol) of
1-hydroxybenzotriazole hydrate, 38.5 mg (0.201 mmol)
1-(3-dimethylaminopropyl)-3-ethylcarbodiimi- de hydrochloride. The
mixture was stirred for 16 h and then concentrated in vacuo. The
residue was taken up in ethyl acetate and washed with water, 0.5 N
hydrochloric acid, washed with sodium bicarbonate, saturated brine,
dried over magnesium sulfate, filtered and concentrated in vacuo.
The residue was purified by low pressure silica gel column
chromatography using a 10% to 35% gradient of ethyl
acetate/CH.sub.2Cl.sub.2 eluent to yield 69.3 mg of the title
product as a white solid. TLC: Rf=0.46, 60% ethyl
acetate/CH.sub.2Cl.sub.2, HPLC: Rt=15.0 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 190
[0975] A. Compound XXXI (A=H, R.sup.3=Methyl, R.sup.3'=Methyl,
D=Isobutyl, E=4-acetamidophenyl, Hydrochloride Salt).
[0976] To a solution of 60.1 mg of the resultant compound of
Example 189B in CH.sub.2Cl.sub.2 (1 mL) at 0.degree. to 5.degree.
C. was added trifluoromethanesulfonic acid (1 mL). After stirring
for 0.75 h, the resultant mixture was concentrated in vacuo and the
resulting white solid used directly for subsequent reaction.
[0977] B. Compound 190.
[0978] To a solution of 37 mg (0.059 mmol) of the resultant
compound of Example 190A in CH.sub.2Cl.sub.2 (3 mL) was added
sequentially, at ambient temperature under an atmosphere of
nitrogen, with 15.4 mg (0.089 mmol) of 1-hydroxybenzotriazole
hydrate, and 17.8 mg (0.089 mmol) EDC. The mixture was stirred for
16 h and then concentrated in vacuo. The residue was taken up in
EtOAc and washed with saturated brine, dried over magnesium
sulfate, filtered and concentrated in vacuo. The residue was
purified by thin layer silica gel column chromatography using 50%
of EtOAc in CH.sub.2Cl.sub.2 as eluent to yield 32.5 mg of the
title product. TLC: Rf=0.35, 50% EtOAc/CH.sub.2Cl.sub.2, HPLC:
Rt=15.65 min; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 191
[0979] A. (2S, 3RS)-S-Amino-1-chloro-2-hydroxy-4-phenylbutane).
[0980] A solution of 2.24 g (6.71 mmol) of (1S,
2RS)-N-(1-benzyl-3-chloro-- 2-hydroxypropyl)-benzyloxycarbonylamine
in 5 mL of methanol was added, at ambient temperature under a
nitrogen atmosphere, to a slurry of 0.22 g (10% by weight) of 10%
palladium on carbon in 60 mL methanol and hydrogenerated for 24 h,
under a slight positive pressure of hydrogen. The mixture was
filtered and concentrated in vacuo to yield 1.34 g of the mixed
diastereomeric products. TLC: Rf=0.33, 10k
CH.sub.3OH/CH.sub.2Cl.su- b.2.
[0981] B. (2S)-2-Benzyloxycarbonylamino-N.sup.1-((1S,
2RS)-1-benzyl-3-chloro-2-hydroxypropyl)-N.sup.4-trityl
Succinamide.
[0982] A solution of 1.34 g (6.71 mmol) of the resultant compounds
of Example 191A in 60 mL of dichloromethane was treated
sequentially, at ambient temperature under an atmosphere of
nitrogen, with 3.58 g (7.05 mmol) of
Cbz-N.sup..delta.-trityl-asparagine, 0.95 g (7.05 mmol) of
1-hydroxybenzotriazole hydrate, 1.35 g (7.05 mmol) of EDC. The
mixture was stirred for 24 hours and then concentrated in vacuo.
The residue was taken up in ethyl acetate and washed with water,
saturated NaHCO.sub.3, saturated NaCl; dried over MgSO.sub.4;
filtered and concentrated in vacuo. The residue was purified by low
pressure silica gel column chromatography using 10% ethyl
acetate/dichloromethane as eluent to yield 3.08 g total of the
mixed diastereomeric products. TLC: Rf=0.75, 0.83, 406
EtOAc/CH.sub.2Cl.sub.2; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
[0983] C. (2S)-2-Amino-N.sup.1-((1S,
2RS)-1-benzyl-3-chloro-2-hydroxypropy- l)-N.sup.4-trityl
Succinamide.
[0984] A solution of 2.80 g (4.06 mmol) of the resultant compounds
of Example 191B in 5 mL of methanol was added, at ambient
temperature under a nitrogen atmosphere, to a slurry of 0.28 g (10%
by weight) of 10% palladium on carbon in 100 mL methanol and
hydrogenated for 24 h under a slight positive pressure of hydrogen.
The mixture was filtered and concentrated in vacuo to yield 2.26 g
of the mixed distereomeric products. TLC: Rf=0.42, 10%
CH.sub.3OH/CH.sub.2Cl.sub.2.
[0985] D. (2S)-2-((1S,
2RS)-1-Benzyl-3-chloro-2-hydroxypropyl)-N.sup.1
((quinoline-2-carbonyl)-amino)-N.sup.4-trityl Succinamide.
[0986] A solution of 2.26 g (4.06 mmol) of the resultant compounds
of Example 191C in 60 mL of dichloromethane was treated
sequentially, at ambient temperature under an atmosphere of
nitrogen, with 0.74 g (4.27 mmol) of quinaldic acid, 0.58 g (4.27
mmol) of 1-hydroxybenzotriazole hydrate, and 0.82 g (4.27 mmol) of
EDC. After 24 hours, 30 mL of dichloromethane was added. The
mixture was washed with water, 5% NaHCO.sub.3 solution, saturated
NaCl, dried over MgSO.sub.4, filtered and concentrated in vacuo.
The residue was dissolved in 50% ethyl acetate/hexane and filtered
through a plug of silica gel. Removal of the solvents yielded 2.30
g of the mixed diastereomeric products. TLC: Rf=0.53, 0.58, 40%
EtOAc/CH.sub.2Cl.sub.2; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
[0987] E.
(2S)-2-((1S,2RS)-1-Benzyl-2-hydroxy-3-iodopropyl)-N.sup.1-((quin-
oline-2-carbonyl)-amino)-N.sup.4-trityl Succinamide.
[0988] A solution of 1.05 g (1.48 mmol) of the resultant compounds
of Example 191D and 0.36 g (2.37 mmol) of sodium iodide in 15 mL of
methyl ethyl ketone was heated to reflux for 24 hours. The mixture
was cooled to room temperature and then concentrated in vacuo. The
residue was taken up in dichloromethane and washed with water,
saturated NaCl, dried over MgSO.sub.4, filtered and concentrated in
vacuo to yield 1.3 g of the mixed diastereomeric products. TLC:
Rf=0.58, 0.65, 40% EtOAc/CH.sub.2Cl.sub.2; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
[0989] F. (2S)-2-((1S, 2 syn,
anti)-3-(2-methylpropyl)amino-1-benzyl-2-hyd-
roxypropyl)-N.sup.1-((quinoline-2-carbonyl)-amino)-N.sup.4-trityl
Succinamide.
[0990] A solution of 207.6 mg (0.26 mmol) of the resultant
compounds of Example 191E and 0.5 mL (5.17 mmol) of isobutylamine
in 9 mL of acetonitrile in a sealed tube was heated to reflux for
24 hours. After cooling to room temperature, the mixture was
concentrated in vacuo. The residue was taken up in dichloromethane
and washed with water, saturated NaCl, dried over MgSO.sub.4,
filtered and concentrated in vacuo to yield 209.2 mg of the mixed
diastereomeric products. TLC: Rf=0.11, 10%
CH.sub.3OH/CH.sub.2Cl.sub.2; (.sup.1H)-NMR (CDCl.sub.3) consistent
with structure.
[0991] G. Compound XIV ((syn, anti) --OH, A=quinoline-2-carbonyl,
D'=Isobutyl).
[0992] A solution of 192.9 mg (0.26 mmol) of the resultant
compounds of Example 191F and 0.07 mL (0.388 mmol) of
diisopropylethylamine in 5 mL of dichloromethane was treated with
112.9 mg (0.517 mmol) of di-tert-butyldicarbonate. After 24 hours,
the mixture was diluted with dichloromethane. The mixture washed
with water, 5k NaHCO.sub.3, 0.5 N HCl, saturated NaCl, dried over
MgSO.sub.4, filtered and concentrated in vacuo. The residue was
purified by low pressure silica gel column chromatography using 40%
ethyl acetate/dichloromethane as eluent to yield 147.3 mg of the
mixed diastereomeric products. TLC: Rf=0.60, 0.67, 40%
EtOAc/CH.sub.2Cl.sub.2; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
[0993] H. Compounds 191.
[0994] A solution of 147.3 mg (0.174 mmol) of the resultant
compounds of Example 191G in 2 mL of dichloromethane was treated
with 2 mL of trifluoroacetic acid. After 4 hours, the mixture was
concentrated in vacuo. TLC: Rf=0.11, 10%
CH.sub.3OH/CH.sub.2Cl.sub.2. To a solution of the resultant
compound in 2 mL of dichloromethane was sequentially added 0.5 mL
of saturated NaHCO.sub.3, small amount of solid NaHCO.sub.3 and 67
mg (0.226 mmol) of a mixture of
4-acetamido-3-fluorobenzenesulphonyl chloride and
3-acetamido-4-fluorobenzenesulphonyl chloride. After 3 hours, the
mixture was diluted with dichloromethane. The two layers were
separated and the aqueous layer was extracted once with
dichloromethane. The combined organic layer was washed with
saturated NaCl then dried over MgSO.sub.4, filtered and
concentrated in vacuo. The residue was purified by low pressure
silica gel column chromatography using 2% methanol/dichloromethane
was eluent to yield 64 mg of the mixed diastereomers and
regioisomers which were further purified with preparative HPLC to
yield 18.9 mg of the mixed regioisomers comprising compounds 191 as
a white solid. TLC: Rf=0.14, 5% CH.sub.3OH/CH.sub.2Cl.su- b.2;
HPLC, Rt=13.36 min; (.sup.1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 193
[0995] Compound 193.
[0996] A solution of 81.2 mg (0.096 mmol) of the resultant lower Rf
diastereomer of Example 9/192A in 3 mL of dichloromethane was
treated with 3 mL of trifluoroacetic acid. After 4 hours, the
mixture was concentrated in vacuo. TLC: Rf=0.11, 10%
CH.sub.3OH/CH.sub.2Cl.sub.2. To a solution of 20.6 mg (0.0431 mmol)
of the resultant residue in 1 mL of dichloromethane was
sequentially added 0.3 mL of saturated NaHCO.sub.3, small amount of
solid NaHCO.sub.3 and 12.4 mg (0.053 mmol) of
4-acetamidobenzenesulphonyl chloride. After 3 hours, the mixture
was diluted with dichloromethane. The two layers were separated and
the aqueous layer was extracted once with dichloromethane. The
combined organic layer was washed with brine then dried over
MgSO.sub.4, filtered and concentrated in vacuo. The residue was
purified by preparative HPLC to yield 8.3 mg of the title compound
as a white solid; TLC: Rf=0.10, 5% CH.sub.3OH/CH.sub.2Cl.sub.2;
HPLC, Rt=12.7 min; (1H)-NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 194
[0997] Compound 194.
[0998] To a solution of 13.0 mg (0.026 mmol) of the trifluoroacetic
acid deprotection product described in Example 193 in 1 mL of
dichloromethane was sequentially added 0.3 mL of saturated
NaHCO.sub.3, small amount of solid NaHCO.sub.3 and 8.4 mg (0.033
mmol) of 5-(isoxazol-3-yl)thiophene-2- -sulphonyl chloride. After 3
hours, the mixture was diluted with dichloromethane. The two layers
were separated and the aqueous layer was extracted once with
dichloromethane. The combined organic layer was washed with brine
then dried over MgSO.sub.4, filtered and concentrated in vacuo. The
residue was purified by preparative HPLC to yield 5.1 mg of the
title product as a white solid; TLC: Rf=0.27, 5%
CH.sub.3OH/CH.sub.2Cl.sub.2; HPLC, Rt=14.4 min; (.sup.1H)-NMR
(CDCl.sub.3) consistent with structure.
EXAMPLE 195
[0999] A. Compound XXII(A=(S)-3-tetrahydrofuryl,
D'=Cyclopentylmethyl, A'=Tert-butoxycarbonyl).
[1000] To a solution of 264 mg of the resultant compound of Example
140D in 10 mL of CH.sub.2Cl.sub.2 was added 0.14 mL of
disopropylethylamine and 175 mg of di-tert butylpyrocarbonate.
After stirring for 4 hours, the mixture was diluted with 50 mL of
CH.sub.2Cl.sub.2, washed with 0.5N of HCl and brine, dried over
magnesium sulfate, filtered and concentrated in vacuo to yield 364
mg of the title compound as a white solid which was used without
subsequent purification. TLC: Rf=0.58, 40%
EtOAc/CH.sub.2Cl.sub.2.
[1001] B. A solution of 334 mg of the resultant compound of Example
195A in 5 mL of ethanol was hydrogenated under 30 psi of hydrogen
in the presence of 80 mg of platinum (IV) oxide for 24 hours. The
mixture was filtered and concentrated. The residue was purified by
a low pressure silica gel column chromatography using 20% EtOAc in
CH.sub.2Cl.sub.2 eluent to yield 268 mg of the title compound. TLC:
Rf=0.55, 40% EtOAc/CH.sub.2Cl.sub.2. (.sup.1H)-NMR (CDCl.sub.3)
consistent with structure.
[1002] C.
[1003] A solution of 268 mg of the resultant compound of Example
195B in 10 mL of EtOAc was treated with anhydrous HCl gas for 5
min. The reaction mixture was sparged with nitrogen then
concentrated in vacuo and the resulting white solid used without
subsequent purification for subsequent reaction.
[1004] D. Compound 195.
[1005] To a solution of 233 mg of the crude resultant compound of
Example 195C in 10 mL of CH.sub.2Cl.sub.2 was added 2 mL of
saturated aqueous sodium bicarbonate and 149 mg of
4-methyloxybenzene sulfonyl chloride. After 3 hours, the resulting
mixture was diluted with CH.sub.2Cl.sub.2, washed with sodium
bicarbonate, brine, dried over magnesium sulfate, filtered and
concentrated in vacuo. The residue was purified by low pressure
silica gel column chromatography using 0% to 20%
EtOAc/CH.sub.2Cl.sub.2 to yield 225 mg of the title compound as a
white solid. TLC: Rf=0.40, 20% EtOAc/CH.sub.2Cl.sub.2; HPLC:
Rt=15.65 min.: (.sup.1H)NMR (CDCl.sub.3) consistent with
structure.
EXAMPLE 196
[1006] A.
(1S,2S)-N-(1-Isobutyl-3-chloro-2-hydroxypropyl)benzyloxycarbonyl-
amine.
[1007] To a solution of N-Cbz-leucine chloromethyl ketone (2.0g) in
20 mL ofmethanol was added, at 0.degree. C., 1.0 g of sodium
borohydride and the mixture was stirred at ambient temperature for
24 h. The solution was concentrated under reduced pressure and the
residue partitioned between 20 mL of saturated aqueous NH.sub.4Cl
and 500 ml of diethyl ether. The organic fraction was separated,
dried over MgSO.sub.4 and concentrated in vacuo and the residue
purified by silica gel chromatography to yield 1.8 g of white
solid.
[1008] B. (1S)-1-1(S)(Carbobenzyloxy)amino-2-isobutyl-oxirane.
[1009] To a solution of the resultant compound of Example 196A (300
mg) in absolute ethanol was added 67 mg of powdered KOH. The
mixture was stirred for 3 h at ambient temperature, filtered
through diatomaceous earth, and concentrated in vacuo. The residue
was dissolved in diethyl ether, dried over MgSO.sub.4, and
concentrated to yield 230 mg of colorless oil, which was used
directly for subsequent reaction.
[1010] C.
(2R,3S)-N.sup.3-Carbobenzyloxy-N.sup.1-isobutyl-1,3-diamino-2-hy-
droxy-5-methylhexane.
[1011] A 230 mg portion of the resultant compound of example 196B
was suspended in 5 mL of isobutylamine and the mixture stirred
overnight at ambient temperature. The mixture was concentrated in
vacuo to yield the title product as 179 mg of a white solid, which
was used directly for subsequent reaction.
[1012] D. Compound I (A=Tert-butoxycarbonyl, x=0,D'=Isobutyl,
E=4-methoxyphenyl, (s)-hydroxy).
[1013] Following the procedure described in Example 81, a solution
of the resultant compound of example 196C (170 mg) in
CH.sub.2Cl.sub.2 was reacted with 4-methoxybenzenxulfoyl chloride
(150 mg) in the presence of aqueous NaHCO.sub.3. Workup and silica
gel chromatography yielded 90 mg of product as a white solid.
[1014] E. Compound I (A=H, x=0, D=Isobutyl, E=4-methoxyphenyl,
(S)-hydroxy).
[1015] A solution of the resultant compound of Example 196D (90 mg)
in ethanol was treated with 50 mg of 10% palladium on carbon and
the mixture stirred under an atmosphere of hydrogen. After
completion of reaction, the mixture was filtered and concentrated
in vacuo to yield 60 mg of the title compound which was used
directly for subsequent reaction.
[1016] F. Compound 196.
[1017] Reaction of the resultant compound of Example 196E (60 mg)
in CH.sub.2Cl.sub.2 was reacted with the resultant product of
example 48A (150 mg) as described earlier yielded, following
aqueous workup, drying over MgSO.sub.4, filtering, and
concentration in vacuo, a residue which was purified by silica gel
chromatography using methanol/CH.sub.2CL.sub.2 as eluant to yield
the title product as 40 mg of white solid.
[.sup.1h]-NMR(CDCl.sub.3) consistent with structure.
EXAMPLE 197
[1018] We measured the inhibition constants of the compounds listed
in Table VII against HIV-1 protease using the above-cited method of
Pennington et al.
[1019] We also measured the anti-viral potency of the compounds in
CCRM-CEM cells by the above-cited method of Meek et al. In the
Tables below, K.sub.i and IC.sub.90 values are expressed in nM.
[1020] In Table VIII, the following classifications have been
employed:
[1021] A: inhibits HIV replication at concentration of 100 nM or
less.
[1022] B: inhibits HIV replication at concentration of between 101
and 1,000 nM.
[1023] C: inhibits HIV replication at a concentration of between
1,001 and 10,000 nM.
[1024] D: inhibits HIV replication at a concentration of between
10,001 and 40,000 nM.
[1025] ND: not tested.
9 TABLE VII Compound K.sub.1 value 1 4.0 2 2.0 3 32 4 19 5 2.0 6
3.0 7 8.0 8 850 9 4.0 10 4.0 11 34 12 0.1 13 0.2 14 0.1 15 <0.1
16 <0.1 17 <0.1 18 <0.1 19 <0.1 20 0.1 21 0.7 22 1.0 23
1.5 24 32,500 25 3,000 26 0.1 27 8.0 28 17 29 17 30 61 31 ND 32 2.5
33 80 34 17 35 4.0 36 19 37 0.1 38 1.5 39 17 40 1,100 41 220 42 46
43 4,200 44 5.0 45 6.0 46 154 47 4.0 48 1.4 49 9.0 50 11 51 ND 52
0.4 53 27 54 22 55 430 56 60 57 200 58 34 59 206 60 4.0 61 4.0 62
72 63 7.0 64 3.0 65 0.7 66 0.4 67 7,400 68 120 69 42 70 25 71 470
72 4000 73 140 74 11 75 290 76 ND 77 ND 78 ND 79 ND 80 ND 81 2.3 82
1.5 83 ND 84 1.4 85 4.0 86 5.0 87 10 88 1.4 89 2.0 90 93 91 2.5 92
20 93 0.8 94 1.7 95 1.3 96 8.0 97 2.5 98 0.5 99 0.24 100 0.16 101
250 102 33 103 4.5 104 5.5 105 7.5 106 1.4 107 1.4 108 2.0 109 6.0
110 28 111 0.3 112 4.0 113 3.0 114 0.35 115 0.5 116 <0.1 117
0.26 118 <0.1 119 1.8 120 11 121 2.0 122 1.2 123 10 124 1.1 125
0.3 126 310 127 650 128 >5000 129 19 130 14 131 60 132 6.0 133
24 134 8.4 135 2.7 136 18 137 26 138 1.4 139 1.2 140 <0.1 141
0.1 142 <0.1 143 <0.1 144 8.0 145 1.4 146 2.0 147 1.6 148 0.2
149 1.7 150 6.0 151 0.8 152 2.5 153 0.2 154 0.5 155 1.7 156 2.8 157
0.7 158 <0.1 159 0.2 160 1.0 161 20 162 0.5 163 0.5 164 130 165
0.4 166 <0.1 167 0.45 168 0.6 169 <0.1 170 0.2 171 0.2 172 21
173 0.6 174 10 175 0.1 176 <0.1 177 <0.1 178 0.1 179 0.4 180
<0.1 181 0.3 182 0.2 183 0.1 184 5.0 185 3.5 186 140 187 0.3 188
11.5 189 5,500 190 ND 191 33 192 67 193 400 194 350 195 0.2 196
ND
[1026]
10 TABLE VIII Compound IC.sub.90 Range 1 C 2 B 3 C 4 C 5 B 6 B 7 D
8 ND 9 B 10 B 11 ND 12 A 13 A 14 A 15 A 16 B 17 B 18 B 19 B 20 A 21
A 22 B 23 B 24 ND 25 ND 26 B 27 C 28 ND 29 C 30 ND 31 ND 32 C 33 ND
34 ND 35 B 36 ND 37 B 38 C 39 C 40 ND 41 ND 42 ND 43 ND 44 B 45 C
46 ND 47 C 48 B 49 C 50 C 51 C 52 B 53 ND 54 C 55 ND 56 ND 57 ND 58
ND 59 ND 60 C 61 C 62 ND 63 C 64 C 65 C 66 B 67 ND 68 ND 69 ND 70
ND 71 ND 72 ND 73 ND 74 ND 75 ND 76 ND 77 ND 78 ND 79 ND 80 ND 81 C
82 C 83 ND 84 C 85 C 86 B 87 C 88 B 89 C 90 ND 91 B 92 ND 93 B 94 B
95 C 96 ND 97 B 98 B 99 B 100 A 101 ND 102 ND 103 C 104 C 105 ND
106 C 107 C 108 C 109 B 110 ND 111 C 112 B 113 B 114 B 115 B 116 A
117 C 118 B 119 C 120 ND 121 C 122 C 123 ND 124 D 125 B 126 ND 127
ND 128 ND 129 ND 130 ND 131 ND 132 ND 133 ND 134 ND 135 C 136 ND
137 ND 138 B 139 B 140 A 141 B 142 A 143 A 144 B 145 B 146 B 147 B
148 A 149 B 150 B 151 C 152 ND 153 ND 154 ND 155 B 156 B 157 B 158
A 159 B 160 A 161 ND 162 C 163 B 164 ND 165 B 166 A 167 B 168 A 169
A 170 B 171 A 172 ND 173 A 174 ND 175 A 176 ND 177 ND 178 ND 179 ND
180 ND 181 ND 182 B 183 B 184 ND 185 ND 186 ND 187 B 188 C 189 ND
190 ND 191 C 192 C 193 ND 194 ND 195 A 196 ND
[1027] As demonstrated in Tables VII and VIII, all of the compounds
tested displayed inhibitory and anti-viral activity. Moreover,
several of these compounds exhibited activity levels far greater
than those of known HIV protease inhibitors.
[1028] While we have described a number of embodiments of this
invention, it is apparent that our basic constructions may be
altered to provide other embodiments which utilize the products and
processes of this invention. Therefore, it will be appreciated that
the scope of this invention is to be defined by the appended
claims, rather than by the specific embodiments which have been
presented by way of example.
* * * * *